Studies of the lethargic (lh/lh) mouse model of absence seizures: regulatory mechanisms and identification of the lh gene.
To understand the cellular and molecular mechanisms that underlie generalized absence seizures sufficiently well to design rational, efficacious new therapies for patients, it is necessary to turn to animal models to gain insights into these mechanisms. The lethargic (lh/lh) mutant mouse expresses spontaneous absence seizures that share behavioral, electrographic, and anticonvulsant profiles with absence seizures in patients. This validates its use to study the mechanisms that underlie absence seizures. This chapter discusses two scientific approaches that involve the use of lh/lh mice. The first part of the chapter discusses neurobiologic approaches used to investigate critical mechanisms that regulate the synchronized burst firing within the thalamocortical network that generates absence seizures. Two of these critical mechanisms have been studied in detail with lh/lh mice. The first critical mechanism involves the required activation of gamma-aminobutyric acid B (GABAB) receptors to generate absence seizures. Because the numbers of GABAB receptors are increased in thalamocortical populations among lh/lh mice compared with littermates without epilepsy, these receptors appear to play a pathophysiologic role in the expression of absence seizures among lh/lh mice. Moreover, there may be a role for GABAB receptors in the generation of absence seizures among humans, because administration of compounds that activate GABAB receptors can produce absence seizures among humans. These findings suggest that GABAB receptor antagonists may represent a new class of antiabsence compounds that will be efficacious against absence seizures among patients. A second critical mechanism that regulates generation of absence seizures involves GABAA receptors in the nucleus reticularis thalami (NRT), a nucleus that sends GABA-ergic afferents to thalamic relay nuclei. Activation of GABAA receptors in the NRT appears to suppress the generation of absence seizures among lh/lh mice and in other models. Moreover, clonazepam may exert its antiabsence actions through this mechanism. Together, these findings suggest that compounds that selectively activate GABAA receptor isoforms expressed in NRT may represent a class of antiabsence drugs that could have fewer side effects than compounds currently used to treat patients. The second part of the chapter discusses a molecular genetic approach to delineation of the mechanisms that underlie absence seizures. Absence seizures among lh/lh mice are caused by a single-gene defect on chromosome 2. If positional cloning and gene isolation techniques are successful, it will be possible to identify the lh disease gene. Subsequent studies of the lh gene product should greatly increase not only our understanding of the pathophysiologic basis for absence seizures among lh/lh mice but also our ability to seek similar mutations in homologous genes in human families that express absence seizures. Accordingly, strategies and progress in cloning and identifying the lh disease gene are presented.


Long-term neurological complications associated with surgery and peripheral nerve blockade: outcomes after 1065 consecutive blocks.
Peripheral nerve blockade is gaining popularity as an analgesic option for both upper or lower limb surgery. Published evidence supports the improved efficacy of regional techniques when compared to conventional opioid analgesia. The incidence of neurological deficit after surgery associated with peripheral nerve block is unclear. This paper reports on neurological outcomes occurring after 1065 consecutive peripheral nerve blocks over a one-year period from a single institution. All patients receiving peripheral nerve blocks for surgery were prospectively followed for up to 12 months to determine the incidence and probable cause of any persistent neurological deficit. Formal independent neurological review and testing was undertaken as indicated. Thirteen patients reported symptoms that warranted further investigation. A variety of probable causes were identified, with peripheral nerve block being implicated in two cases (one resolved at nine months and one remaining persistent). Overall incidence of block-related neuropathy was 0.22%. Persistent postoperative neuropathy is a rare but serious complication of surgery associated with peripheral nerve block. Formal follow-up of all such blocks is recommended to assess causality and allow for early intervention.


Xylella fastidiosa esterase rather than hydroxynitrile lyase.
In 2009, we reported that the product of the gene SCJ21.16 (XFa0032) from Xylella fastidiosa, a xylem-restricted plant pathogen that causes a range of diseases in several important crops, encodes a protein (XfHNL) with putative hydroxynitrile lyase activity. Sequence analysis and activity tests indicated that XfHNL exhibits an α/β-hydrolase fold and could be classified as a member of the family of FAD-independent HNLs. Here we provide a more detailed sequence analysis and new experimental data. Using pure heterologously expressed XfHNL we show that this enzyme cannot catalyse the cleavage/synthesis of mandelonitrile and that this protein is in fact a non-enantioselective esterase. Homology modelling and ligand docking simulations were used to study the active site and support these results. This finding could help elucidate the common ancestor of esterases and hydroxynitrile lyases with an α/β -hydrolase fold.


An oncogenic phenoscape of colonic stem cell polarization.
Cancer cells are regulated by oncogenic mutations and microenvironmental signals, yet these processes are often studied separately. To functionally map how cell-intrinsic and cell-extrinsic cues co-regulate cell fate, we performed a systematic single-cell analysis of 1,107 colonic organoid cultures regulated by (1) colorectal cancer (CRC) oncogenic mutations, (2) microenvironmental fibroblasts and macrophages, (3) stromal ligands, and (4) signaling inhibitors. Multiplexed single-cell analysis revealed a stepwise epithelial differentiation phenoscape dictated by combinations of oncogenes and stromal ligands, spanning from fibroblast-induced Clusterin (CLU)+ revival colonic stem cells (revCSCs) to oncogene-driven LRIG1+ hyper-proliferative CSCs (proCSCs). The transition from revCSCs to proCSCs is regulated by decreasing WNT3A and TGF-β-driven YAP signaling and increasing KRASG12D or stromal EGF/Epiregulin-activated MAPK/PI3K flux. We find that APC loss and KRASG12D collaboratively limit access to revCSCs and disrupt stromal-epithelial communication-trapping epithelia in the proCSC fate. These results reveal that oncogenic mutations dominate homeostatic differentiation by obstructing cell-extrinsic regulation of cell-fate plasticity.


Metal accumulation in the polychaete Hediste japonica with emphasis on interaction between heavy metals and petroleum hydrocarbons.
The accumulation of cadmium (Cd) and copper (Cu) in the polychaete Hediste japonica exposed to the mixture of Cd (or Cu) and petroleum hydrocarbons (PHCs) was investigated and compared with that exposed to single Cd (or Cu). The increased bioavailability of Cd or Cu with exposure concentrations resulted in an increase in the accumulation and net accumulation rate of Cd or Cu during single metal exposure. The net accumulation rate of Cd increased, but the net accumulation rate of Cu decreased with exposure time during single metal exposure, suggesting that H. japonica could actively regulate Cu burden in their body by inhibition of absolute uptake or promotion of excretion. The interactions between Cd (or Cu) and PHCs had complicated influences on the net accumulation rate of Cd and Cu in H. japonica under the condition of the binary mixture, which are dependent on their concentration combinations and exposure time.


Comparative effects of single- and linear triple-site rapid bipolar pacing on atrial activation in canine models.
Nonuniform conduction may cause block and/or delay, thereby providing a substrate for the onset and maintenance of reentrant atrial arrhythmias. We tested the hypothesis that linear triple-site, bipolar, rapid pacing (LTSBRP) of the right atrium generates more uniform wave-front propagation compared with single-site, bipolar, rapid pacing (SSBRP), thereby reducing and/or eliminating conduction block and delay that is otherwise present. Five dogs with pericarditis and three normal dogs were studied. Three plunge-wire electrode pairs were placed 5-7 mm apart in both perpendicular and parallel configurations at the superior aspect of the crista terminalis and were used to pace at 200- and 300-ms cycle lengths for < or =6 s. During pacing, 380 electrograms were recorded simultaneously from electrode arrays placed epicardially on the atria, which produced activation sequence maps for each pacing episode. Local conduction-velocity vectors were computed for each site during each episode. Histograms of absolute velocity vector angles from the x-axis (of the crista terminalis) were plotted to assess uniformity of wave-front propagation, and the magnitude of each vector was computed to assess the local speed. LTSBRP showed 1) more uniform linear activation wave fronts compared with SSBRP, 2) velocity vectors with a more uniform magnitude and direction compared with SSBRP, 3) a predominant absolute velocity vector angle vs. a scattered angle distribution with SSBRP, and 4) shorter right atrial activation time and faster mean epicardial speed than SSBRP for each pacing cycle length. LTSBRP created a more uniform wave-front propagation with less or no conduction block and/or delay compared with SSBRP.


Inhibition of TFF3 Enhances Sensitivity-and Overcomes Acquired Resistance-to Doxorubicin in Estrogen Receptor-Positive Mammary Carcinoma.
Dose-dependent toxicity and acquired resistance are two major challenges limiting the efficacious treatment of mammary carcinoma (MC) with doxorubicin. Herein, we investigated the function of Trefoil Factor 3 (TFF3) in the sensitivity and acquired resistance of estrogen receptor positive (ER+) MC cells to doxorubicin. Doxorubicin treatment of ER+MC cells increased TFF3 expression. The depletion of TFF3 by siRNA or inhibition with a small molecule TFF3 inhibitor (AMPC) synergistically enhanced the efficacy of doxorubicin in ER+MC through the suppression of doxorubicin-induced AKT activation and enhancement of doxorubicin-induced apoptosis. Elevated expression of TFF3 and increased activation of AKT were also observed using a model of acquired doxorubicin resistance in ER+MC cells. AMPC partially re-sensitized the doxorubicin resistant cells to doxorubicin-induced apoptosis. Indeed, doxorubicin resistant ER + MC cells exhibited increased sensitivity to AMPC as a single agent compared to doxorubicin sensitive cells. In vivo, AMPC attenuated growth of doxorubicin sensitive ER+MC xenografts whereas it produced regression of xenografts generated by doxorubicin resistant ER+MC cells. Hence, TFF3 inhibition may improve the efficacy and reduce required doses of doxorubicin in ER+MC. Moreover, inhibition of TFF3 may also be an effective therapeutic strategy to eradicate doxorubicin resistant ER+MC.


The development of a highly photostable and chemically stable zwitterionic near-infrared dye for imaging applications.
A novel zwitterionic near-infrared (NIR) dye, ZWCC, has been designed and synthesized. It shows significantly enhanced photostability and chemical stability compared to the existing zwitterionic NIR dye. In addition, the feasibility of labeling ZWCC with biological ligands was investigated and used in live cell imaging applications.


Large-bore tunneled central venous catheter insertion in patients with coagulopathy.
To evaluate the safety of tunneled central venous catheter (TCVC) insertion in patients with an International Normalized Ratio (INR) of greater than or equal to 1.5 or a platelet count lower than 50,000/dL. Our division's criteria for acceptable coagulation parameters in patients undergoing TCVC insertion is INR no greater than 2 and platelet count of at least 25,000/dL. Three-thousand one-hundred and eighty-eight TCVCs (8-14.5 F) placed between July 2001 and July 2008 were identified. After excluding incomplete records, 2,514 patients undergoing 3,170 TCVC placements were identified. The most current platelet count and INR at the time of TCVC insertion were identified. Bleeding complications were compared between patients with platelet counts lower than 50,000/dL and/or INR of at least 1.5 and published standards for complication rates for TCVC insertion. Of the total of 3,170 TCVCs placed, 428 were in patients whose most current platelet count was lower than 50,000/dL and 361 were in patients with an INR of 1.5 or greater. After exclusion of intercurrent blood product transfusion, 626 TCVCs placed in 567 patients with a platelet count lower than 50,000/dL and/or an INR of at least 1.5 were identified. There were 27 complications in the entire study population (0.85%); three were in patients with platelet counts lower than 50,000/dL and/or INR of at least 1.5. There were three bleeding complications in the study population (0.095%), none in patients with platelet count lower than 50,000/dL and/or INR of at least 1.5. Placement of TCVCs in patients with a platelet count between 25,000/dL and 50,000/dL and/or an INR between 1.5 and 2 is safe even without coagulation product transfusions.


Undistorted X-ray Absorption Spectroscopy Using s-Core-Orbital Emissions.
Detection of secondary emissions, fluorescence yield (FY), or electron yield (EY), originating from the relaxation processes upon X-ray resonant absorption has been widely adopted for X-ray absorption spectroscopy (XAS) measurements when the primary absorption process cannot be probed directly in transmission mode. Various spectral distortion effects inherent in the relaxation processes and in the subsequent transportation of emitted particles (electron or photon) through the sample, however, undermine the proportionality of the emission signals to the X-ray absorption coefficient. In the present study, multiple radiative (FY) and nonradiative (EY) decay channels have been experimentally investigated on a model system, FeCl3 aqueous solution, at the excitation energy of the Fe L-edge. The systematic comparisons between the experimental spectra taken from various decay channels, as well as the comparison with the theoretically simulated Fe L-edge XA spectrum that involves only the absorption process, indicate that the detection of the Fe 3s → 2p partial fluorescence yield (PFY) gives rise to the true Fe L-edge XA spectrum. The two key characteristics generalized from this particular decay channel-zero orbital angular momentum (i.e., s orbital) and core-level emission-set a guideline for obtaining undistorted X-ray absorption spectra in the future.


Validity of ICD-9-CM codes for breast, lung and colorectal cancers in three Italian administrative healthcare databases: a diagnostic accuracy study protocol.
Administrative healthcare databases are useful tools to study healthcare outcomes and to monitor the health status of a population. Patients with cancer can be identified through disease-specific codes, prescriptions and physician claims, but prior validation is required to achieve an accurate case definition. The objective of this protocol is to assess the accuracy of International Classification of Diseases Ninth Revision-Clinical Modification (ICD-9-CM) codes for breast, lung and colorectal cancers in identifying patients diagnosed with the relative disease in three Italian administrative databases. Data from the administrative databases of Umbria Region (910,000 residents), Local Health Unit 3 of Napoli (1,170,000 residents) and Friuli--Venezia Giulia Region (1,227,000 residents) will be considered. In each administrative database, patients with the first occurrence of diagnosis of breast, lung or colorectal cancer between 2012 and 2014 will be identified using the following groups of ICD-9-CM codes in primary position: (1) 233.0 and (2) 174.x for breast cancer; (3) 162.x for lung cancer; (4) 153.x for colon cancer and (5) 154.0-154.1 and 154.8 for rectal cancer. Only incident cases will be considered, that is, excluding cases that have the same diagnosis in the 5 years (2007-2011) before the period of interest. A random sample of cases and non-cases will be selected from each administrative database and the corresponding medical charts will be assessed for validation by pairs of trained, independent reviewers. Case ascertainment within the medical charts will be based on (1) the presence of a primary nodular lesion in the breast, lung or colon-rectum, documented with imaging or endoscopy and (2) a cytological or histological documentation of cancer from a primary or metastatic site. Sensitivity and specificity with 95% CIs will be calculated. Study results will be disseminated widely through peer-reviewed publications and presentations at national and international conferences.


Development of a herpes simplex virus subunit glycoprotein vaccine for prophylactic and therapeutic use.
A herpes simplex virus subunit vaccine has been tested for immunogenicity and protective efficacy in animal models. The vaccine is a mixture of the viral glycoproteins gB and gD expressed in mammalian cells as secreted carboxyl terminal truncated derivatives. The antigens have been combined with several adjuvants, including a lipophilic derivative of a muramyl tripeptide, MTP-PE, and administered in a stable, low-oil emulsion. Titers of antibody that are threefold to 15-fold greater than those obtained with alum and within twofold of those generated with complete Freund's adjuvant are elicited in animals, including primates. Prophylactic immunization of guinea pigs provides nearly complete protection against intravaginal challenge with herpes simplex virus type 2. Treatment of previously infected guinea pigs reduces the frequency and severity of recurrent disease, although variations in efficacy dependent on the antigen, adjuvant, and dosing regimen employed are observed. The immunogenicity of this vaccine in animals provides the rationale for further testing in human clinical trials.


Interfacial regions governing internal cavities of dendrimers. Studies of poly(alkyl aryl ether) dendrimers constituted with linkers of varying alkyl chain length.
This report deals with a study of the properties of internal cavities of dendritic macromolecules that are capable of encapsulating and mediating photoreactions of guest molecules. The internal cavity structures of dendrimers are determined by the interfacial regions between the aqueous exterior and hydrocarbon like interior constituted by the linkers that connect symmetrically sited branch points constituting the dendrimer and head groups that cap the dendrimers. Phloroglucinol-based poly(alkyl aryl ether) dendrimers constituted with a homologous series of alkyl linkers were undertaken for the current study. Twelve dendrimers within first, second, and third generations, having ethyl, n-propyl, n-butyl, and n-pentyl groups as the linkers and hydroxyl groups at peripheries in each generation, were synthesized. Encapsulation of pyrene and coumarins by aqueous basic solutions of dendrimers were monitored by UV-vis and fluorescence spectroscopies, which showed that a lower generation dendrimer with an optimal alkyl linker presented better encapsulation abilities than a higher generation dendrimer. Norrish type I photoreaction of dibenzyl ketone was carried out within the above series of dendrimers to probe their abilities to hold guests and reactive intermediate radical pairs within themselves. The extent of cage effect from the series of third generation dendrimers was observed to be higher with dendrimers having an n-pentyl group as the linker.


Age-related Trends in Adults with Urinary Tract Infections Presenting to the Emergency Department: A 5-Year Experience.
Urinary tract infections (UTIs) are among the most common bacterial infections, affecting 150 million people worldwide each year. Importantly, the incidence of UTI increases markedly with age. The increasing resistance to empirically prescribed antimicrobial agents complicates the management of this disease. This makes UTI an important issue in increasingly aging population and updated epidemiological investigation is advisable. To understand the epidemiological variation in UTI resistance patterns among differently aged populations, we conducted a retrospective study among patients presenting to the emergency department (ED) of a large tertiary-care hospital in Italy during January 2013 and June 2017. 1281 patients who presented with UTI symptoms to the ED, were stratified into four age categories: young adults (18-44 years old;), adults (45-64), the elderly (65-84), and the oldest old (≥ 85). Inclusion criteria were urine collected in ED patients with UTI symptoms and first positive culture from one given patient in a given year. 362 (28.2%) patients had a urine culture with positive result, leading to a total of 459 germs isolated, stratified into four categories: young adults (58 isolates, 12.6%), adults (98, 21.4%), the elderly (174, 37.9%), and the oldest old (129, 28.1%). Escherichia coli represents the 60% of all monomicrobial infections, followed in frequency by Klebsiella pneumoniae (15%), and Enterococcus faecalis (5%). The other 20% of the infections are caused by various germs. The most common association of germs in polymicrobial is E. coli + E. faecalis, accounting for the 28% of all infections. Overall, we found a peak of susceptibility to amoxicillin (AMX) in the oldest old ( 81%), significantly higher compared to young adults (54%), adults (47%) and elderly (35%) (p<0,001). For ciprofloxacin (CIP) there is a greater susceptibility in the young adult (55.5%), but not so marked compared to the other three groups; for fosfomicin (FOS) the susceptibility was greater in the group of adults (60%) compared to young adults, elderly and the oldest old. Also for trimethoprim/ sulfamethoxazole (TMP-SMX) we found greater susceptibility in the adult group (60%), followed by the oldest old (57,6%), young adults (49%) and elderly (47%). Age-related differences in antimicrobial-resistant microorganisms were evident for adults with UTI, and could potentially contribute to the risk of inappropriate empirical therapy in elderly patients. Thus, different empirical antimicrobial regimens should be considered for distinct age groups.


Development of High Dose Oseltamivir Phosphate Dry Powder for Inhalation Therapy in Viral Pneumonia.
Oseltamivir phosphate (OP) is an antiviral drug available only as oral therapy for the treatment of influenza and as a potential treatment option when in combination with other medication in the fight against the corona virus disease (COVID-19) pneumonia. In this study, OP was formulated as a dry powder for inhalation, which allows drug targeting to the site of action and potentially reduces the dose, aiming a more efficient therapy. Binary formulations were based on micronized excipient particles acting like diluents, which were blended with the drug OP. Different excipient types, excipient ratios, and excipient size distributions were prepared and examined. To investigate the feasibility of delivering high doses of OP in a single dose, 1:1, 1:3, and 3:1 drug/diluent blending ratios have been prepared. Subsequently, the aerosolization performance was evaluated for all prepared formulations by cascade impaction using a novel medium-resistance capsule-based inhaler (UNI-Haler). Formulations with micronized trehalose showed relatively excellent aerosolization performance with highest fine-particle doses in comparison to examined lactose, mannitol, and glucose under similar conditions. Focusing on the trehalose-based dry-powder inhalers' (DPIs) formulations, a physicochemical characterization of extra micronized grade trehalose in relation to the achieved performance in dispersing OP was performed. Additionally, an early indication of inhaled OP safety on lung cells was noted by the viability MTT assay utilizing Calu-3 cells.


Quantitative determination of joint incongruity and pressure distribution during simulated gait and cartilage thickness in the human hip joint.
The objective of this study was to provide quantitative data on hip-joint incongruity and pressure during a simulated walking cycle and on articular-cartilage thickness in the same set of specimens. Using a casting technique in eight specimens of the human hip (age: 18-75 years), we determined the width of the joint space (incongruity) required at minimal load for contact at four phases of the gait cycle. The pressure distribution, measured with pressure-sensitive film, was determined at physiologic load magnitudes on the basis of in vivo measurements of hip-joint forces. Cartilage thickness was assessed with A-mode ultrasound. At minimal loading, the average maximum width of the joint space ranged from 1.1 to 1.5 mm in the acetabular roof, with the contact areas located ventro-superiorly and dorso-inferiorly throughout the gait cycle. At physiological loading, the width decreased and the contact areas covered the complete articular surface during midstance and heel-off but not during heel-strike or toe-off. The pressure distribution was inhomogeneous during all phases, with average maximum pressures of 7.7 +/- 1.95 MPa at midstance. The cartilage thickness varied considerably throughout the joint surfaces; maxima greater than 3 mm were found ventro-superiorly. These data can be used to generate and validate computer models to determine the load-sharing between the interstitial fluid and the solid proteoglycan-collagen matrix of articular cartilage, the latter being relevant for the initiation of mechanically induced cartilage degeneration.


Ocular and endocrine function in patients with pituitary tumors. Operative results following transnasal, transsphenoidal approach with marsupialization of the sella turcica.
Thirty-one patients with pituitary tumors underwent transphenoidal operation via a unilateral, trans-nasal technique preserving the nasal septum and marsupializing the sella turcica unless CSF leak could be demonstrated. These two modifications neither detracted from exposure nor predisposed to delayed complications and significantly facilitated the procedure.


Prevalence of cataract and pseudophakia/aphakia among adults in the United States.
To determine the prevalence of cataract and pseudophakia/aphakia in the United States and to project the expected change in these prevalence figures by 2020. Summary prevalence estimates of cataract and of pseudophakia/aphakia were prepared separately for black, white, and Hispanic persons (for whom only cataract surgery data were available) in 5-year age intervals starting at 40 years for women and men. The estimates were based on a standardized definition of various types of cataract: cortical, greater than 25% of the lens involved; posterior subcapsular, present according to the grading system used in each study; and nuclear, greater than or equal to the penultimate grade in the system used. Data were collected from major population-based studies in the United States, and, where appropriate, Australia, Barbados, and Western Europe. The age-, gender-, and race/ethnicity-specific rates were applied to 2000 US Census data, and projected population figures for 2020, to obtain overall estimates. An estimated 20.5 million (17.2%) Americans older than 40 years have cataract in either eye, and 6.1 million (5.1%) have pseudophakia/aphakia. Women have a significantly (odds ratio = 1.37; 95% confidence interval, 1.26-1.50) higher age-adjusted prevalence of cataract than men in the United States. The total number of persons who have cataract is estimated to rise to 30.1 million by 2020; and for those who are expected to have pseudophakia/aphakia, to 9.5 million. The number of Americans affected by cataract and undergoing cataract surgery will dramatically increase over the next 20 years as the US population ages.


A World Health Organization field trial assessing a proposed ICD-11 framework for classifying patient safety events.
To assess the utility of the proposed World Health Organization (WHO)'s International Classification of Disease (ICD) framework for classifying patient safety events. Independent classification of 45 clinical vignettes using a web-based platform. The WHO's multi-disciplinary Quality and Safety Topic Advisory Group. The framework consists of three concepts: harm, cause and mode. We defined a concept as 'classifiable' if more than half of the raters could assign an ICD-11 code for the case. We evaluated reasons why cases were nonclassifiable using a qualitative approach. Harm was classifiable in 31 of 45 cases (69%). Of these, only 20 could be classified according to cause and mode. Classifiable cases were those in which a clear cause and effect relationship existed (e.g. medication administration error). Nonclassifiable cases were those without clear causal attribution (e.g. pressure ulcer). Of the 14 cases in which harm was not evident (31%), only 5 could be classified according to cause and mode and represented potential adverse events. Overall, nine cases (20%) were nonclassifiable using the three-part patient safety framework and contained significant ambiguity in the relationship between healthcare outcome and putative cause. The proposed framework enabled classification of the majority of patient safety events. Cases in which potentially harmful events did not cause harm were not classifiable; additional code categories within the ICD-11 are one proposal to address this concern. Cases with ambiguity in cause and effect relationship between healthcare processes and outcomes remain difficult to classify.


Sampling sites for detection of feline herpesvirus-1, feline calicivirus and Chlamydia felis in cats with feline upper respiratory tract disease.
Feline herpesvirus-1 (FHV-1), feline calicivirus (FCV) and Chlamydia felis are involved in feline upper respiratory tract disease (FURTD). Clinical signs caused by these agents can overlap, and the involvement of certain pathogens is often unpredictable. The objectives of this study were to compare detection rates of FHV-1, FCV and C felis at different sampling sites, and to investigate the correlation between positive test results and clinical signs in cats with FURTD. Swabs were taken from the nose, pharynx, tongue and conjunctiva of 104 cats with signs of FURTD. Real-time PCR was performed on all samples for the detection of FHV-1, FCV and C felis. Infectious agents were identified in 93 (89.4%) cats. Of these, 55.8% were positive for FHV-1, 50.0% for FCV and 35.6% for C felis. FCV was found more frequently in the oropharynx (92.3% of FCV-positive cats) and on the tongue (90.4%) than the conjunctiva (38.5%) (P <0.001). There was no significant difference between the four sampling sites for the detection of FHV-1 and C felis. If nasal samples had also been taken, 94.9% of FHV-1-positive cats, 96.2% of FCV-positive cats and 81.1% of C felis-positive cats would have been detected. The oropharynx can be recommended as the preferred single sampling site for the detection of FCV, FHV-1 and C felis if only one sample can be taken; however, taking samples at different sites significantly increases the detection rate for all pathogens studied. Interestingly, sampling from a site with FURTD-associated lesions did not increase the likelihood of detecting the infectious agents.


Visualizing Deep Learning Models for the Detection of Referable Diabetic Retinopathy and Glaucoma.
Convolutional neural networks have recently been applied to ophthalmic diseases; however, the rationale for the outputs generated by these systems is inscrutable to clinicians. A visualization tool is needed that would enable clinicians to understand important exposure variables in real time. To systematically visualize the convolutional neural networks of 2 validated deep learning models for the detection of referable diabetic retinopathy (DR) and glaucomatous optic neuropathy (GON). The GON and referable DR algorithms were previously developed and validated (holdout method) using 48 116 and 66 790 retinal photographs, respectively, derived from a third-party database (LabelMe) of deidentified photographs from various clinical settings in China. In the present cross-sectional study, a random sample of 100 true-positive photographs and all false-positive cases from each of the GON and DR validation data sets were selected. All data were collected from March to June 2017. The original color fundus images were processed using an adaptive kernel visualization technique. The images were preprocessed by applying a sliding window with a size of 28 × 28 pixels and a stride of 3 pixels to crop images into smaller subimages to produce a feature map. Threshold scales were adjusted to optimal levels for each model to generate heat maps highlighting localized landmarks on the input image. A single optometrist allocated each image to predefined categories based on the generated heat map. Visualization regions of the fundus. In the GON data set, 90 of 100 true-positive cases (90%; 95% CI, 82%-95%) and 15 of 22 false-positive cases (68%; 95% CI, 45%-86%) displayed heat map visualization within regions of the optic nerve head only. Lesions typically seen in cases of referable DR (exudate, hemorrhage, or vessel abnormality) were identified as the most important prognostic regions in 96 of 100 true-positive DR cases (96%; 95% CI, 90%-99%). In 39 of 46 false-positive DR cases (85%; 95% CI, 71%-94%), the heat map displayed visualization of nontraditional fundus regions with or without retinal venules. These findings suggest that this visualization method can highlight traditional regions in disease diagnosis, substantiating the validity of the deep learning models investigated. This visualization technique may promote the clinical adoption of these models.


[Role of biochemical risk factors and markers for the risk of atherosclerosis].
The prevalence of atherosclerotic cardiovascular disorders is constantly increasing worldwide, and the aging of the population will make this trend rather unlikely to be reversed. It is a diffuse, systemic, chronic disorder that starts early in childhood and progresses asymptomatically throughout the adult life. Later in life, it is clinically manifested as coronary artery disease, stroke, transient ischemic attack and peripheral occlusive arterial disease. The notions on the pathogenesis have been almost revolutionized over the past decades and over 300 risk factors have been associated with genesis, progression and complication of this challenging disorder. Particularly, biochemical risk factors and markers play a crucial role in all steps of atherosclerosis process, and entail a great relevance in risk stratification and early prevention.


Roles of Forkhead-box Transcription Factors in Controlling Development, Pathogenicity, and Stress Response in Magnaporthe oryzae.
Although multiple transcription factors (TFs) have been characterized via mutagenesis to understand their roles in controlling pathogenicity and infection-related development in Magnaporthe oryzae, the causal agent of rice blast, if and how forkhead-box (FOX) TFs contribute to these processes remain to be characterized. Four putative FOX TF genes were identified in the genome of M. oryzae, and phylogenetic analysis suggested that two of them (MoFKH1 and MoHCM1) correspond to Ascomycota-specific members of the FOX TF family while the others (MoFOX1 and MoFOX2) are Pezizomycotina-specific members. Deletion of MoFKH1 (ΔMofkh1) resulted in reduced mycelial growth and conidial germination, abnormal septation and stress response, and reduced virulence. Similarly, ΔMohcm1 exhibited reduced mycelial growth and conidial germination. Conidia of ΔMofkh1 and ΔMohcm1 were more sensitive to one or both of the cell cycle inhibitors hydroxyurea and benomyl, suggesting their role in cell cycle control. On the other hand, loss of MoFOX1 (ΔMofox1) did not show any noticeable changes in development, pathogenicity, and stress response. Deletion of MoFOX2 was not successful even after repeated attempts. Taken together, these results suggested that MoFKH1 and Mo-HCM1 are important in fungal development and that MoFKH1 is further implicated in pathogenicity and stress response in M. oryzae.


RNA Polymerase II Dependent Crosstalk between H4K16 Deacetylation and H3K56 Acetylation Promotes Transcription of Constitutively Expressed Genes.
Nucleosome dynamics in the coding region of a transcriptionally active locus is critical for understanding how RNA polymerase II progresses through the gene body. Histone acetylation and deacetylation critically influence nucleosome accessibility during DNA metabolic processes like transcription. Effect of such histone modifications is context and residue dependent. Rather than effect of individual histone residues, the network of modifications of several histone residues in combination generates a chromatin landscape that is conducive for transcription. Here we show that in Saccharomyces cerevisiae, crosstalk between deacetylation of the H4 N-terminal tail residue H4K16 and acetylation of the H3 core domain residue H3K56, promotes RNA polymerase II progression through the gene body. Results indicate that deacetylation of H4K16 precedes and in turn induces H3K56 acetylation. Effectively, recruitment of Rtt109, the HAT responsible for H3K56 acetylation is essentially dependent on H4K16 deacetylation. In Hos2 deletion strains, where H4K16 deacetylation is abolished, both H3K56 acetylation and RNA polymerase II recruitment gets significantly impaired. Notably, H4K16 deacetylation and H3K56 acetylation are found to be essentially dependent on active transcription. In summary, H4K16 deacetylation promotes H3K56 acetylation and the two modifications together work towards successful functioning of RNA polymerase II during active transcription.


Role of ornithine decarboxylase in enterocyte mitochondrial function and integrity.
We examined the role of ornithine decarboxylase (ODC) and polyamine biosynthesis in regulating mitochondrial function and integrity along the crypt-villus axis in male Sprague-Dawley rats. Isolated villus tip enterocytes from control rats demonstrated a greater cellular capacity for glucose oxidation than crypt enterocytes. Mitochondrial enzyme activities were similar along the crypt-villus axis. The role of ODC was assessed by treating experimental rats with the irreversible ODC inhibitor alpha-difluoromethylornithine (DFMO) for 24 h. Animals receiving DFMO demonstrated a decreased CO2 production from [2-(14)C]pyruvate along the entire crypt-villus axis coupled with an increase in lactate production in the upper cell populations. CO2 production from [14C]glucose and total ATP levels were not affected by DFMO treatment. Ultrastructural examination revealed localized mitochondrial swelling and bursting only in enterocytes corresponding to the population of cells newly emerged from the crypt during DFMO treatment. In DFMO-treated animals, 2 microM spermine completely prevented the structural mitochondrial injury and restored the metabolic crypt-villus gradient. These results suggest that as enterocytes migrate from the crypt up the villus, mitochondrial function increases to handle the increased metabolic demands placed on the cell by nutrient absorption. ODC activity and polyamines are necessary for this increased mitochondrial function and have a role in the maintenance of mitochondrial integrity in maturing enterocytes migrating from the crypt onto the villus.


Chromosome-scale comparative sequence analysis unravels molecular mechanisms of genome dynamics between two wheat cultivars.
Recent improvements in DNA sequencing and genome scaffolding have paved the way to generate high-quality de novo assemblies of pseudomolecules representing complete chromosomes of wheat and its wild relatives. These assemblies form the basis to compare the dynamics of wheat genomes on a megabase scale. Here, we provide a comparative sequence analysis of the 700-megabase chromosome 2D between two bread wheat genotypes-the old landrace Chinese Spring and the elite Swiss spring wheat line 'CH Campala Lr22a'. Both chromosomes were assembled into megabase-sized scaffolds. There is a high degree of sequence conservation between the two chromosomes. Analysis of large structural variations reveals four large indels of more than 100 kb. Based on the molecular signatures at the breakpoints, unequal crossing over and double-strand break repair were identified as the molecular mechanisms that caused these indels. Three of the large indels affect copy number of NLRs, a gene family involved in plant immunity. Analysis of SNP density reveals four haploblocks of 4, 8, 9 and 48 Mb with a 35-fold increased SNP density compared to the rest of the chromosome. Gene content across the two chromosomes was highly conserved. Ninety-nine percent of the genic sequences were present in both genotypes and the fraction of unique genes ranged from 0.4 to 0.7%. This comparative analysis of two high-quality chromosome assemblies enabled a comprehensive assessment of large structural variations and gene content. The insight obtained from this analysis will form the basis of future wheat pan-genome studies.


AOASM Position Statement on Esports, Active Video Gaming, and the Role of the Sports Medicine Physician.
Electronic sports, or esports, has a global audience of over 300 million fans and is increasing in popularity, resulting in projected revenue of over $1 billion by the end of this past year. The global pandemic of 2020 had little to no effect on these increasing numbers because athletes have been able to continue to engage in sports because of its electronic nature and fans have been able to follow them virtually. Esports has been recognized as an organized sport by the International Olympic Committee, the US National Collegiate Athletic Association, and several secondary school athletic associations within the United States. In addition, professional teams have been established in several major cities within the United States, Canada, Europe, and Australia. With the growth of esports, the necessity of incorporating esports medicine into the practice of sports medicine physicians has become paramount. Esports can be played on a monitor or screen and played using physical activity in what has become known as active video gaming. Within both of these platforms, there have emerged certain conditions unique to esports. There are also certain conditions seen in other sports applicable to esports athletes. This document will review the evaluation of the esports athlete, introduce conditions unique to these athletes and review common conditions seen in esports, discuss diagnostics used in the evaluation of esports athletes, introduce treatment options for conditions unique to esports and review those for commonly seen injuries in esports, discuss prevention of injuries in esports, and introduce a framework for the future development of esports medicine that can be introduced into the daily practice of the sports medicine physician.


Induction of Maternal Immune Activation in Mice at Mid-gestation Stage with Viral Mimic Poly(I:C).
Maternal immune activation (MIA) model is increasingly well appreciated as a rodent model for the environmental risk factor of various psychiatric disorders. Numerous studies have demonstrated that MIA model is able to show face, construct, and predictive validity that are relevant to autism and schizophrenia. To model MIA, investigators often use viral mimic polyinosinic:polycytidylic acid (poly(I:C)) to activate the immune system in pregnant rodents. Generally, the offspring from immune activated dam exhibit behavioral abnormalities and physiological alterations that are associated with autism and schizophrenia. However, poly(I:C) injection with different dosages and at different time points could lead to different outcomes by perturbing brain development at different stages. Here we provide a detailed method of inducing MIA by intraperitoneal (i.p.) injection of 20 mg/kg poly(I:C) at mid-gestational embryonic 12.5 days (E12.5). This method has been shown to induce acute inflammatory response in the maternal-placental-fetal axis, which ultimately results in the brain perturbations and behavioral phenotypes that are associated with autism and schizophrenia.


Non-BRCA1/2 Breast Cancer Susceptibility Genes: A New Frontier with Clinical Consequences for Plastic Surgeons.
Twenty percent of breast cancer cases may be related to a genetic mutation conferring an increased risk of malignancy. The most common and prominent breast cancer susceptibility genes are BRCA1 and BRCA2, found in nearly 40% of such cases. However, continued interest and investigation of cancer genetics has led to the identification of a myriad of different breast cancer susceptibility genes. Additional genes, each with unique significance and associated characteristics, continue to be recognized. Concurrently, advanced genetic testing, while still controversial, has become more accessible and cost-effective. As oncologic and reconstructive advances continue to be made in prophylactic breast reconstructive surgery, patients may present to plastic surgeons with an increasingly more diverse array of genetic diagnoses to discuss breast reconstruction. It is therefore imperative that plastic surgeons be familiar with these breast cancer susceptibility genes and their clinical implications. We, therefore, aim to review the most common non-BRCA1/2 breast cancer susceptibility genetic mutations in an effort to assist plastic surgeons in counseling and managing this unique patient population. Included in this review are syndromic breast cancer susceptibility genes such as TP53, PTEN, CDH1, and STK11, among others. Nonsyndromic breast cancer susceptibility genes herein reviewed include PALB2, CHEK2, and ataxia telangiectasia mutated gene. With this knowledge, plastic surgeons can play a central role in the diagnosis and comprehensive treatment, including successful breast reconstruction, of all patients carrying genetic mutations conferring increased risk for breast malignancies.


The admissions process in a graduate-entry dental school: can we predict academic performance?
To assess the association between the admissions performance and subsequent academic achievement within a graduate-entry dental school. The study was conducted at the University of Aberdeen Dental School. UCAS forms for course applicants were reviewed and assigned a pre-admission score (PAS) and a tariff given for the UCAS personal statement (UCAS). Individuals ranked highest were invited to attend multiple mini-interviews (MMI), which were scored. Data was correlated with academic performance reported as the University Common Assessment Scale (0-20). Comparisons were also made between the first degree and subsequent educational achievement. Data were analysed by multiple linear regression, Pearson correlation and unstacked ANOVA (IBM SPSS Statistics 19). Data were obtained for 75 students (F: 50; M: 25). A correlation between performance at MMI and CAS scores was identified (r = 0.180, p = 0.001, df = 538). A correlation was also noted between each student's first degree and the CAS scores (F = 4.08, p = 0.001, df = 9). This study suggests that candidate performance at MMI might be a stronger predictor of academic and clinical performance of graduate-entry dental students compared to other pre-interview selection criteria. The first degree for such a programme also appears to be significant.


Acute Systemic Complications of Convulsive Status Epilepticus-A Systematic Review.
Status epilepticus is a neurologic emergency with high morbidity and mortality requiring neurointensive care and treatment of systemic complications. This systematic review compiles the current literature on acute systemic complications of generalized convulsive status epilepticus in adults and their immediate clinical impact along with recommendations for optimal neurointensive care. We searched PubMed, Medline, Embase, and the Cochrane library for articles published between 1960 and 2016 and reporting on systemic complications of convulsive status epilepticus. All identified studies were screened for eligibility by two independent reviewers. Key data were extracted using standardized data collection forms. Thirty-two of 3,046 screened articles were included. Acute manifestations and complications reported in association with generalized convulsive status epilepticus can affect all organ systems fueling complex cascades and multiple organ interactions. Most reported complications result from generalized excessive muscle contractions that increase body temperature and serum potassium levels and may interfere with proper and coordinated function of respiratory muscles followed by hypoxia and respiratory acidosis. Increased plasma catecholamines can cause a decay of skeletal muscle cells and cardiac function, including stress cardiomyopathy. Systemic complications are often underestimated or misinterpreted as they may mimic underlying causes of generalized convulsive status epilepticus or treatment-related adverse events. Management of generalized convulsive status epilepticus should center on the administration of antiseizure drugs, treatment of the underlying causes, and the attendant systemic consequences to prevent secondary seizure-related injuries. Heightened awareness, systematic clinical assessment, and diagnostic workup and management based on the proposed algorithm are advocated as they are keys to optimal outcome.


Constructing computational pipelines.
Many bioinformatics tasks involve creating a computational pipeline from existing software components and algorithms. The traditional approach is to glue components together using scripts written in a programming language such as Perl. However, a new, more powerful approach is emerging that promises to revolutionise the way bioinformaticians create applications from existing components, an approach based on the concept of the scientific workflow. Scientific workflows are created in graphical environments known as workflow management systems. They have many benefits over traditional program scripts, including speed of development, portability, and their suitability for developing complex, distributed applications. This chapter explains how to design and implement bioinformatics workflows using free, Open Source software tools, such as the Taverna workflow management system. We also demonstrate how new and existing tools can be deployed as Web services so that they can be easily integrated into novel computational pipelines using the scientific workflow paradigm.


Patient-level factors associated with hemoglobin A1C testing in Alabama Medicaid beneficiaries with diabetes.
We evaluated patient-level factors associated with receipt of hemoglobin A1c (HbA1c) testing among Alabama Medicaid beneficiaries with type 2 diabetes. We conducted a retrospective analysis of person-year observations from Medicaid claims data from 2011 to 2020. Adults aged 19-64 years with type 2 diabetes and continuous enrollment in Medicaid for study year and year prior were included. Primary outcomes were ≥ 1 and ≥ 2 HbA1c test(s) per year. We conducted multivariable Poisson regression stratified by Medicaid eligibility reason (disability, poverty) examining the association of study year, demographics, clinical factors, and healthcare utilization with HbA1c testing. We analyzed 288,379 observations, 51% with disability-based, 49% poverty-based eligibility. Overall, 57% observations had ≥ 1 HbA1c, 35% had ≥ 2 HbA1c tests. More observations with disability-based than poverty-based eligibility had ≥ 1 (76% vs. 38%) and ≥ 2 HbA1c tests (49% vs. 20%). Patient-level factors were associated with a higher likelihood of having ≥ 1 HbA1c: Black race and older age (disability-based eligibility); year after 2011, female sex, and younger age (poverty-based eligibility); and rurality, insulin use, endocrinology care, diabetes complications, and ambulatory care visits (both groups). Just over one-third of adult Alabama Medicaid beneficiaries with diabetes had ≥ 2 HbA1c tests per year; testing frequency differed by Medicaid eligibility.


Adrenal incidentaloma in adults - management recommendations by the Polish Society of Endocrinology.
A wide use of imaging techniques results in more frequent diagnosis of adrenal incidenataloma. To analyse the current state of knowledge on adrenal incidentaloma in adults in order to prepare practical management recommendations. Following a discussion, the Polish Society of Endocrinology expert working group have analysed the available data and summarised the analysis results in the form of recommendations. Unenhanced adrenal computed tomography (CT) may be recommended as an initial assessment examination helpful in the differentiation between adenomas and "non-adenomatous" lesions. In the case of density > 10 Hounsfield units, CT with contrast medium washout assessment or MRI are recommended. However, in all patients with adrenal incidentaloma, hormonal assessment is recommended in order to exclude pheochromocytoma and hypercortisolism, notwithstanding the clinical picture or concomitant diseases. In addition, examination to exclude primary hyperaldosteronism is suggested in patients with diagnosed hypertension or hypokalaemia. Surgical treatment should be recommended in patients with adrenal incidentaloma, where imaging examinations suggest a malignant lesion (oncological indication) or with confirmed hormonal activity (endocrinological indication). The basis of the surgical treatment is laparoscopic adrenalectomy. Patients with suspected pheochromocytoma must be pharmacologically prepared prior to surgery. In patients not qualified for surgery, control examinations (imaging and laboratory tests) should be established individually, taking into consideration such features as the size, image, and growth dynamics of the tumour, clinical symptoms, hormonal tests results, and concomitant diseases.


Diagnosis and Clinical Management of Amyotrophic Lateral Sclerosis and Other Motor Neuron Disorders.
This article reviews the clinical features, diagnostic pathway, therapies, and current understanding of the pathophysiology of amyotrophic lateral sclerosis (ALS). The spectrum of motor neuron diseases is reviewed, and the clinical heterogeneity of ALS is described. ALS is increasingly recognized as a clinical spectrum disorder with pure upper and pure lower motor neuron presentations, supported by genetic links. The phenotypic variability is broad. Identification of ALS-related genes provides insights into disease mechanisms. ALS is a progressive fatal multisystem neurodegenerative disease primarily affecting motor neurons. Clinical recognition of suspicious symptoms and the appropriate laboratory evaluation are essential to limit diagnostic delay and avoid unnecessary testing and procedures. ALS has broad genetic and hypothesized environmental causes and phenotypic variability. Recognizing related motor neuron diseases will prevent misdiagnosis while allowing proper disease counseling. Although ALS cannot be cured, implementation of appropriate symptomatic treatment is valuable.


Lidocaine blockage of basolateral potassium channels in the amphibian urinary bladder.
1. Basolateral membranes of the frog urinary bladder were investigated after increasing the cationic conductance of the apical membrane by the incorporation of nystatin. 2. K+ currents were recorded in the presence of a mucosa to serosa oriented K+ gradient (SO4(2-) Ringer solution). Nystatin caused a rapid rise of the short-circuit current (Isc) followed by a slow increase over a period of 1-2 h. 3. Impedance analysis showed that the apical membrane resistance was drastically reduced by nystatin. The slow increase in Isc was accompanied by a progressive increase in basolateral conductance. 4. The transepithelial current and conductance recorded in the presence of nystatin could be depressed with lidocaine added to the mucosal and serosal solution. The effects of lidocaine were completely reversible. 5. Noise analysis showed that lidocaine induced additional fluctuations in Isc. The spectrum of these fluctuations was of the Lorentzian type. This noise component is caused by the random interruption of the current through the basolateral K+ channels. The Lorentzian parameters were used to calculate the microscopic parameters of the basolateral K+ channels.


Engineering Symmetry-Selective Couplings of a Superconducting Artificial Molecule to Microwave Waveguides.
Tailoring the decay rate of structured quantum emitters into their environment opens new avenues for nonlinear quantum optics, collective phenomena, and quantum communications. Here, we demonstrate a novel coupling scheme between an artificial molecule comprising two identical, strongly coupled transmon qubits and two microwave waveguides. In our scheme, the coupling is engineered so that transitions between states of the same (opposite) symmetry, with respect to the permutation operator, are predominantly coupled to one (the other) waveguide. The symmetry-based coupling selectivity, as quantified by the ratio of the coupling strengths, exceeds a factor of 30 for both waveguides in our device. In addition, we implement a Raman process activated by simultaneously driving both waveguides, and show that it can be used to coherently couple states of different symmetry in the single-excitation manifold of the molecule. Using that process, we implement frequency conversion across the waveguides, mediated by the molecule, with efficiency of about 95%. Finally, we show that this coupling arrangement makes it possible to straightforwardly generate spatially separated Bell states propagating across the waveguides. We envisage further applications to quantum thermodynamics, microwave photodetection, and photon-photon gates.


Cancer as an ecomolecular disease and a neoplastic consortium.
Current anticancer paradigms largely target driver mutations considered integral for cancer cell survival and tumor progression. Although initially successful, many of these strategies are unable to overcome the tremendous heterogeneity that characterizes advanced tumors, resulting in the emergence of resistant disease. Cancer is a rapidly evolving, multifactorial disease that accumulates numerous genetic and epigenetic alterations. This results in wide phenotypic and molecular heterogeneity within the tumor, the complexity of which is further amplified through specific interactions between cancer cells and the tumor microenvironment. In this context, cancer may be perceived as an "ecomolecular" disease that involves cooperation between several neoplastic clones and their interactions with immune cells, stromal fibroblasts, and other cell types present in the microenvironment. This collaboration is mediated by a variety of secreted factors. Cancer is therefore analogous to complex ecosystems such as microbial consortia. In the present article, we comment on the current paradigms and perspectives guiding the development of cancer diagnostics and therapeutics and the potential application of systems biology to untangle the complexity of neoplasia. In our opinion, conceptualization of neoplasia as an ecomolecular disease is warranted. Advances in knowledge pertinent to the complexity and dynamics of interactions within the cancer ecosystem are likely to improve understanding of tumor etiology, pathogenesis, and progression. This knowledge is anticipated to facilitate the design of new and more effective therapeutic approaches that target the tumor ecosystem in its entirety.


Physical properties of Tld, Sog, Tsg and Dpp protein interactions are predicted to help create a sharp boundary in Bmp signals during dorsoventral patterning of the Drosophila embryo.
Dorsal cell fate in Drosophila embryos is specified by an activity gradient of Decapentaplegic (Dpp), a homologue of bone morphogenetic proteins (Bmps) 2/4. Previous genetic and biochemical studies have revealed that the Sog, Tsg and Tld proteins modify Dpp activity at the post-transcriptional level. The predominant view is that Sog and Tsg form a strong ternary complex with Dpp that prevents it from binding to its cognate receptors in lateral regions of the embryo, while in the dorsal-most cells Tld is proposed to process Sog and thereby liberate Dpp for signaling. In this model, it is not readily apparent how Tld activity is restricted to the dorsal-most cells as it is expressed throughout the entire dorsal domain. In this study, additional genetic and biochemical assays were developed to further probe the relationships between the Sog, Tsg, Tld and Dpp proteins. Using cell based assays, we find that the dynamic range over which Dpp functions for signaling is the same range in which Dpp stimulates the cleavage of Sog by Tld. In addition, our data supports a role for Tsg in sensitizing the patterning mechanism to low levels of Dpp. We propose that the strong Dpp concentration dependence exhibited by the processing reaction, together with movement of Dpp by Sog and Tsg protein can help explain how Tld activity is confined to the dorsal-most region of the embryo through formation of a spatially dependent positive and negative reinforcement loop. Such a mechanism also explains how a sharp rather than smooth signaling boundary is formed.


Optimal ankle axis position for articulated boots.
An articulated boot design is commonly used in skiing and skating sports because it allows sagittal plane ankle mobility while still providing critical frontal plane stability. Although articulated boots have been in use for several decades, current manufacturers of these boots differ in their articulation placement. In this study we determined an optimal position of the ankle articulation axis. We also calculated the amount of anterior skin movement that a boot tongue must account for during a full range of ankle motion. Three-dimensional kinematic data were collected and analyzed from 40 participants moving their right foot through a full range of sagittal plane motion. The calculated horizontal position of the articulation axis was found to be highly predictable from foot length (r = 0.87, standard error of estimate = 3.44 mm), while its vertical component displayed less predictability (r = 0.49, standard error of estimate = 7.46 mm). The expansion required by the boot tongue had a moderate association with foot length and low variability (r = 0.58, standard error of estimate = 0.07 mm). An accurate axis placement will minimize relative motion between the boot cuff and the ankle, reducing friction and motion resistance. An expandable tongue will accommodate full plantar flexion and reduce pressure on the anterior ankle during dorsiflexion, eliminating common pressure-related injuries.


[Production and standardization of Paracoccidioides brasiliensis, Histoplasma capsulatum and Aspergillus fumigatus antigens to be used in immunodiagnosis. Comparison between immunodiffusion and immunoelectrophoresis].
Soluble antigens (Ag) from Paracoccidioides brasiliensis, Histoplasma capsulatum and Aspergillus fumigatus were prepared and standardized by double immunodiffusion (DID) and immunoelectroosmophoresis (IEOP). No difference in sensitivity was observed between the two techniques; 100% of standard patient sera were positive with P. brasiliensis and A. fumigatus Ag and 83.3% were positive with H. capsulatum Ag. The specificity of the tests was verified testing 96 sera from patients with paracoccidioidomycosis, histoplasmosis, systemic candidiasis, sporotrichosis, tuberculosis, lung cancer, visceral or cutaneous leishmaniasis and 18 sera from healthy individuals. All the three antigens were 100% specific with the DID (using the identification pattern indicated by the confluence of test serum with standard serum precipitin lines as a positive criterium). However in the IEOP, the specificity varied with each Ag. Positive reactions with P. brasiliensis Ag were observed in 16.7% of histoplasmosis sera and in 10% of cutaneous leishmaniasis sera. On the other hand 31.8% of paracoccidioidomycosis and 10% of cutaneous leishmaniasis sera reacted with H. capsulatum Ag. The high sensitivity and specificity of the DID test, its easy reproducibility and low cost, led us to consider it highly appropriate as a routine procedure for the screening of patients with respiratory infections.


Personal trust increases cooperation beyond general trust.
In this paper we present a new methodology which, while allowing for anonymous interaction, it also makes possible to compare decisions of cooperating or defecting when playing games within a group, according to whether or not players personally trust each other. The design thus goes beyond standard approaches to the role of trust in fostering cooperation, which is restricted to general trust. It also allows considering the role of the topology of the social network involved may play in the level of cooperation found. The results of this work support the idea that personal trust promotes cooperation beyond the level of general trust. We also found that this effect carries over to the whole group, making it more cohesive, but that higher levels of cohesion rely on a particular topology. As a conclusion, we hypothesize that personal trust is a psychological mechanism evolved to make human social life possible in the small groups our ancestors lived in, and that this mechanism persists and plays a role in sustaining cooperation and social cohesion.


Immunomodulation by vasoactive intestinal peptide is associated with increased survival and growth of Salmonella Typhimurium in mice.
Studies have shown that administration of vasoactive intestinal peptide (VIP) in mice rescues them from lethal endotoxaemia and that this is correlated with decreased concentration of inflammatory cytokines. VIP has, therefore, been proposed as a novel anti-inflammatory which could be used in the treatment of Gram negative sepsis. However, the effect of VIP has not been reported in mice infected with viable Gram negative bacteria. Here, we show that Salmonella enterica serovar Typhimurium 4/74 significantly increased expression of mRNA of a type 1 receptor (VPAC1) for anti-inflammatory vasoactive intestinal peptide (VIP) in murine ileum and mesenteric lymph nodes at day 6 post-infection (d6 pi) and in the spleen at d3 pi. When VIP (5 nmol/ml) was administered to S. Typhimurium-infected mice, there was a significant increase in the number of S. Typhimurium cultured from murine faeces and ileum at d3 and 6 pi and in MLN and spleen at d3 dpi, compared to faeces and tissues examined from mice infected with S. Typhimurium (without VIP administration). Administration of VIP to S. Typhimurium-infected mice also altered the splenic architecture, resulting in a lack of discernable periarterial lymphoid sheaths or marginal zones at d6 pi but liver histology appeared similar on both d3 and d6 pi. The effects of VIP administration were correlated with a significant decrease in expression of inflammatory cytokine mRNA, associated with systemic inflammatory response syndrome (SIRS) of bacteraemia and acute sepsis. We conclude that VIP inhibits expression of diagnostic/prognostic cytokine biomarkers of sepsis in S. Typhimurium-infected mice. However, this occurred with a concomitant increase in Salmonella growth in tissues and increased bacterial shedding in faeces. Thus, VIP may have potential as an adjunctive therapy to antibiotics in sepsis.


Morning cortisol Levels in preschool-aged foster children: differential effects of maltreatment type.
Maltreated foster children are subjected to a range of early adverse experiences, including neglect, abuse, and multiple caregiver disruptions. Research suggests that such disturbances alter the development and subsequent functioning of the hypothalamic-pituitary-adrenocortical system. The current study was designed to investigate morning cortisol levels in 117 foster children and 60 low-income, nonmaltreated children. Maltreatment and foster care placement experiences were coded from official records. Analyses revealed that the foster children were significantly more likely than the nonmaltreated children to have low morning cortisol levels. Additionally, specific maltreatment experiences were significantly associated with the foster children's morning cortisol levels. Foster children with low morning cortisol levels experienced more severe physical neglect than the other foster children. In contrast, foster children with high morning cortisol levels experienced more severe emotional maltreatment. These results suggest that specific early adverse experiences have differential effects on the functioning of the hypothalamic-pituitary-adrenocortical system.


Green tea and the risk of gastric cancer in Japan.
Although laboratory experiments and case-control studies have suggested that the consumption of green tea provides protection against gastric cancer, few prospective studies have been performed. In January 1984, a total of 26,311 residents in three municipalities of Miyagi Prefecture, in northern Japan (11,902 men and 14,409 women 40 years of age or older), completed a self-administered questionnaire that included questions about the frequency of consumption of green tea. During 199,748 person-years of follow-up, through December 1992, we identified 419 cases of gastric cancer (in 296 men and 123 women). We used Cox regression to estimate the relative risk of gastric cancer according to the consumption of green tea. Green-tea consumption was not associated with the risk of gastric cancer. After adjustment for sex, age, presence or absence of a history of peptic ulcer smoking status, alcohol consumption, other dietary elements, and type of health insurance, the relative risks associated with drinking one or two, three or four, and five or more cups of green tea per day, as compared with less than one cup per day, were 1.1 (95 percent confidence interval, 0.8 to 1.6), 1.0 (95 percent confidence interval, 0.7 to 1.4), and 1.2 (95 percent confidence interval, 0.9 to 1.6), respectively (P for trend=0.13). The results were similar after the 117 cases of gastric cancer that were diagnosed in the first three years of follow-up had been excluded, with respective relative risks of 1.2 (95 percent confidence interval, 0.8 to 1.8) 1.0 (95 percent confidence interval, 0.7 to 1.5), and 1.4 (95 percent confidence interval, 1.0 to 1.9) (P for trend=0.07). In a population-based, prospective cohort study in Japan, we found no association between green-tea consumption and the risk of gastric cancer.


Birdsong fails to support object categorization in human infants.
Recent evidence reveals a precocious link between language and cognition in human infants: listening to their native language supports infants' core cognitive processes, including object categorization, and does so in a way that other acoustic signals (e.g., time-reversed speech; sine-wave tone sequences) do not. Moreover, language is not the only signal that confers this cognitive advantage: listening to vocalizations of non-human primates also supports object categorization in 3- and 4-month-olds. Here, we move beyond primate vocalizations to clarify the breadth of acoustic signals that promote infant cognition. We ask whether listening to birdsong, another naturally produced animal vocalization, also supports object categorization in 3- and 4-month-old infants. We report that listening to zebra finch song failed to confer a cognitive advantage. This outcome brings us closer to identifying a boundary condition on the range of non-linguistic acoustic signals that initially support infant cognition.


Prospective memory and subjective memory decline: A neuropsychological indicator of memory difficulties in community-dwelling older people.
Prospective memory difficulties are known to occur in Alzheimer's disease, and may provide an early indicator of cognitive decline. Older people reporting high levels of subjective memory decline (SMD) but without evidence of cognitive decline on standard neuropsychological tests are increasingly considered at increased risk for Alzheimer's disease. Therefore, the objective of this study was to investigate whether prospective memory performance is differentially impaired in older people reporting high levels of SMD as compared to a control group. A total of 195 community-dwelling older adults (Mage = 73.48 years) were assessed for self-reported complaints of memory decline and allocated to either a group reporting high levels of SMD (SMD, n = 96) or a healthy control group (HC, n = 99). Groups were assessed on neuropsychological tests, an experimental prospective memory task (focal vs. nonfocal cue conditions), and a naturalistic prospective memory task. The groups did not differ in performance on standard neuropsychological tests of working memory, executive attention, and episodic retrospective memory. Furthermore, on an experimental task of prospective memory (the Supermarket Shopping Trip task), although performance of both groups was better when cues for prospective memory were focal to the ongoing activity (η2 = .35), the SMD group were not impaired relative to the control group. On a naturalistic prospective memory task, however, there was a small but significant effect, with the SMD group performing more poorly than the HC group (η2 = .02). In older adults with high levels of SMD, naturalistic measures of prospective memory provide an approach to assessing memory performance that can offer a means of investigating the memory complaints of people with SMD. Identifying prospective memory difficulties in SMD also offers a focus for intervention.


Early social deprivation impairs pair bonding and alters serum corticosterone and the NAcc dopamine system in mandarin voles.
Early life stress has a long-term negative impact on emotion, learning, memory and adult sexual behavior, and these deficits most likely impair pair bonding. Here, we investigated whether early social deprivation (ED) affects the formation of pair bonds in socially monogamous mandarin voles (Microtus mandarinus). In a partner preference test (PPT), ED-reared adult females and males did not show a preference for their partner, spent more time exploring the cage of an unfamiliar animal and directed high levels of aggression toward unfamiliar animals. In social interaction test, ED increased exploring behavior only in females, but increased movement around the partner and reduced inactivity in both males and females. Three days of cohabitation did not alter serum corticosterone levels in ED-reared males, but increased corticosterone levels in males that received bi-parental care (PC). Interestingly, serum corticosterone levels in ED- and PC-reared females declined after cohabitation. ED significantly increased basal serum corticosterone levels in males, but had no effect on females. ED significantly up-regulated the levels of dopamine and the mRNA expression of dopamine 1-type receptor (D1R) in the nucleus accumbens (NAcc) in females and males. ED suppressed dopamine 2-type receptor mRNA (D2R) expression in females, but increased this in males. After three days of cohabitation, levels of D1R mRNA and D2R mRNA expression changed in opposite directions in PC-reared voles, but in the same direction in ED-reared males, and only the expression of D2R mRNA increased in ED-reared females. Our results indicate that early social deprivation inhibits pair bonding at adulthood. This inhibition is possibly associated with sex-specific alterations in serum corticosterone, levels of dopamine and mRNA expression of two types of dopamine receptors in the NAcc.


Agreement between Two Methods of Dietary Data Collection in Male Adolescent Academy-Level Soccer Players.
Collecting accurate and reliable nutritional data from adolescent populations is challenging, with current methods providing significant under-reporting. Therefore, the aim of the study was to determine the accuracy of a combined dietary data collection method (self-reported weighed food diary, supplemented with a 24-h recall) when compared to researcher observed energy intake in male adolescent soccer players. Twelve Academy players from an English Football League club participated in the study. Players attended a 12 h period in the laboratory (08:00 h-20:00 h), during which food and drink items were available and were consumed ad libitum. Food was also provided to consume at home between 20:00 h and 08:00 h the following morning under free-living conditions. To calculate the participant reported energy intake, food and drink items were weighed and recorded in a food diary by each participant, which was supplemented with information provided through a 24-h recall interview the following morning. Linear regression, limits of agreement (LOA) and typical error (coefficient of variation; CV) were used to quantify agreement between observer and participant reported 24-h energy intake. Difference between methods was assessed using a paired samples t-test. Participants systematically under-reported energy intake in comparison to that observed (p < 0.01) but the magnitude of this bias was small and consistent (mean bias = -88 kcal·day(-1), 95% CI for bias = -146 to -29 kcal·day(-1)). For random error, the 95% LOA between methods ranged between -1.11 to 0.37 MJ·day(-1) (-256 to 88 kcal·day(-1)). The standard error of the estimate was low, with a typical error between measurements of 3.1%. These data suggest that the combined dietary data collection method could be used interchangeably with the gold standard observed food intake technique in the population studied providing that appropriate adjustment is made for the systematic under-reporting common to such methods.


Assessment of left ventricular mass changes after arteriovenous fistula surgical banding in end-stage renal disease.
Left ventricular hypertrophy (LVH) is a multifactorial complication frequently seen in patients with advanced chronic kidney disease. An arteriovenous fistula (AVF) is the preferred method for hemodialysis access. Once functional, AVFs demonstrate better patency rates and fewer complications when compared to other forms of vascular access. AVFs have been implicated in cardiac remodeling, but it is controversial whether those changes can be reversed by surgical ligation or blood flow reduction. In this study, we describe a cohort of asymptomatic patients with LVH who underwent AVF banding with a two-dimensional-echocardiogram done before and after the intervention to evaluate the association between AVF surgical banding and left ventricular mass (LVM) changes. Our results show that AVF surgical banding did not alter the left ventricular mass index (LVMI) with a mean prebanding LVMI of 70.3 ± 57.5 g/m2 and mean postbanding LVMI of 81.9 ± 55.9 g/m2, (P = 0.4). Our study shows that AVF flow reduction by surgical banding did not alter LVMI, and therefore LVH, in end-stage renal disease patients who have not yet shown clinical manifestations of cardiac disease.


Teleost fish larvae adapt to dietary arachidonic acid supply through modulation of the expression of lipid metabolism and stress response genes.
Dietary fatty acid supply can affect stress response in fish during early development. Although knowledge on the mechanisms involved in fatty acid regulation of stress tolerance is scarce, it has often been hypothesised that eicosanoid profiles can influence cortisol production. Genomic cortisol actions are mediated by cytosolic receptors which may respond to cellular fatty acid signalling. An experiment was designed to test the effects of feeding gilthead sea-bream larvae with four microdiets, containing graded arachidonic acid (ARA) levels (0·4, 0·8, 1·5 and 3·0 %), on the expression of genes involved in stress response (steroidogenic acute regulatory protein, glucocorticoid receptor and phosphoenolpyruvate carboxykinase), lipid and, particularly, eicosanoid metabolism (hormone-sensitive lipase, PPARα, phospholipase A2, cyclo-oxygenase-2 and 5-lipoxygenase), as determined by real-time quantitative PCR. Fish fatty acid phenotypes reflected dietary fatty acid profiles. Growth performance, survival after acute stress and similar whole-body basal cortisol levels suggested that sea-bream larvae could tolerate a wide range of dietary ARA levels. Transcription of all genes analysed was significantly reduced at dietary ARA levels above 0·4 %. Nonetheless, despite practical suppression of phospholipase A2 transcription, higher leukotriene B4 levels were detected in larvae fed 3·0 % ARA, whereas a similar trend was observed regarding PGE2 production. The present study demonstrates that adaptation to a wide range of dietary ARA levels in gilthead sea-bream larvae involves the modulation of the expression of genes related to eicosanoid synthesis, lipid metabolism and stress response. The roles of ARA, other polyunsaturates and eicosanoids as signals in this process are discussed.


[Microbiological diagnosis of epiglottitis].
Forty patients, between July 1977 and April 1982, with clinical diagnosis of epiglottitis were studied for the presence of capsular antigen by counter-immunoelectrophoresis (CIE) and for positive blood culture. Blood culture was positive, for Haemophilus influenzae only, in 15 patients. Haemophilus influenzae type b antigen was present in blood and/or urine of 25 patients (blood 14 patients, urine 18 patients). CIE associated with blood culture give conclusive proofs of Haemophilus influenzae etiology in 30 patients.


ATP1A1-mediated Src signaling inhibits coronavirus entry into host cells.
In addition to transporting ions, the multisubunit Na(+),K(+)-ATPase also functions by relaying cardiotonic steroid (CTS)-binding-induced signals into cells. In this study, we analyzed the role of Na(+),K(+)-ATPase and, in particular, of its ATP1A1 α subunit during coronavirus (CoV) infection. As controls, the vesicular stomatitis virus (VSV) and influenza A virus (IAV) were included. Using gene silencing, the ATP1A1 protein was shown to be critical for infection of cells with murine hepatitis virus (MHV), feline infectious peritonitis virus (FIPV), and VSV but not with IAV. Lack of ATP1A1 did not affect virus binding to host cells but resulted in inhibited entry of MHV and VSV. Consistently, nanomolar concentrations of the cardiotonic steroids ouabain and bufalin, which are known not to affect the transport function of Na(+),K(+)-ATPase, inhibited infection of cells with MHV, FIPV, Middle East respiratory syndrome (MERS)-CoV, and VSV, but not IAV, when the compounds were present during virus inoculation. Cardiotonic steroids were shown to inhibit entry of MHV at an early stage, resulting in accumulation of virions close to the cell surface and, as a consequence, in reduced fusion. In agreement with an early block in infection, the inhibition of VSV by CTSs could be bypassed by low-pH shock. Viral RNA replication was not affected when these compounds were added after virus entry. The antiviral effect of ouabain could be relieved by the addition of different Src kinase inhibitors, indicating that Src signaling mediated via ATP1A1 plays a crucial role in the inhibition of CoV and VSV infections. Coronaviruses (CoVs) are important pathogens of animals and humans, as demonstrated by the recent emergence of new human CoVs of zoonotic origin. Antiviral drugs targeting CoV infections are lacking. In the present study, we show that the ATP1A1 subunit of Na(+),K(+)-ATPase, an ion transporter and signaling transducer, supports CoV infection. Targeting ATP1A1 either by gene silencing or by low concentrations of the ATP1A1-binding cardiotonic steroids ouabain and bufalin resulted in inhibition of infection with murine, feline, and MERS-CoVs at an early entry stage. Infection with the control virus VSV was also inhibited. Src signaling mediated by ATP1A1 was shown to play a crucial role in the inhibition of virus entry by ouabain and bufalin. These results suggest that targeting the Na(+),K(+)-ATPase using cardiotonic steroids, several of which are FDA-approved compounds, may be an attractive therapeutic approach against CoV and VSV infections.


[Regulatory peptides. A functional continuum].
A hypothesis postulating the regulatory peptides continuum is presented. At certain combinations, the numerous regulatory peptides (RP) make up a continuous set of bioactivity spectra, i.e., functionally continuous integrity (continuum) which provides a way for any complex influences on organism functions. Nevertheless, each RP retains its qualitative differences as compared to other RPs. Data on reciprocal influences of RP on their release into body fluids are generalized. The experimental results testify to complex chain reactions of the whole RP continuum in response to changes in any single peptide concentration.


'I never really left the university:' Continuity amongst male academics in the transition from work to retirement.
This study investigates the transition to retirement of male academics. It applies approaches drawn from Cumulative Advantage and Disadvantage (CAD) principles to expand Continuity theory by examining evidence for continuity and inequalities in the lifestyles of the respondents. Biographical-Narrative interviews with 20 retired academics in Brazil and the United Kingdom were conducted and analysed using Thematic Analysis. An exploration of the same occupational group in different cultural and social contexts was carried out to capture a diversity of CAD influences on retirement outcomes. The experience of continuity was individualised to each participant due to their unique combination of advantages, and their subjective interpretation of their experiences. Cumulative processes identified in this study related to (1) occupational roles played in their career; and (2) organisational level policies and practices for retirement. At the level of individual retired men, cultural and social context factors were not perceived as influential in the achievement of continuity in retirement. Continuity theory is still supported, but alone offers limited explanations of the diversity of experiences in the transition to retirement of male academics. A consideration of the role of cumulative processes and systemic dynamics, including how individuals respond to their experiences in retirement, shows how these different factors interact and affect retirement and ageing. Further studies should investigate the processes identified with other occupations, as well as women and minority groups.


Kisspeptin and energy balance in reproduction.
Kisspeptin is vital for the neuroendocrine regulation of GNRH secretion. Kisspeptin neurons are now recognized as a central pathway responsible for conveying key homeostatic information to GNRH neurons. This pathway is likely to mediate the well-established link between energy balance and reproductive function. Thus, in states of severely altered energy balance (either negative or positive), fertility is compromised, as is Kiss1 expression in the arcuate nucleus. A number of metabolic modulators have been proposed as regulators of kisspeptin neurons including leptin, ghrelin, pro-opiomelanocortin (POMC), and neuropeptide Y (NPY). Whether these regulate kisspeptin neurons directly or indirectly will be discussed. Moreover, whether the stimulatory role of leptin on reproduction is mediated by kisspeptin directly will be questioned. Furthermore, in addition to being expressed in GNRH neurons, the kisspeptin receptor (Kiss1r) is also expressed in other areas of the brain, as well as in the periphery, suggesting alternative roles for kisspeptin signaling outside of reproduction. Interestingly, kisspeptin neurons are anatomically linked to, and can directly excite, anorexigenic POMC neurons and indirectly inhibit orexigenic NPY neurons. Thus, kisspeptin may have a direct role in regulating energy balance. Although data from Kiss1r knockout and WT mice found no differences in body weight, recent data indicate that kisspeptin may still play a role in food intake and glucose homeostasis. Thus, in addition to regulating reproduction, and mediating the effect of energy balance on reproductive function, kisspeptin signaling may also be a direct regulator of metabolism.


Interplay between AAUAAA and the trans-splice site in processing of a Caenorhabditis elegans operon pre-mRNA.
About half of Caenorhabditis elegans genes have a 1-2 bp mismatch to the canonical AAUAAA hexamer that signals 3' end formation. One rare variant, AGUAAA, is found at the 3' end of the mai-1 gene, the first gene in an operon also containing gpd-2 and gpd-3. When we expressed this operon under heat shock control, 3' end formation dependent on the AGUAAA was very inefficient, but could be rescued by a single bp change to create a perfect AAUAAA. When AGUAAA was present, most 3' ends formed at a different site, 100 bp farther downstream, right at the gpd-2 trans-splice site. Surprisingly, 3' end formation at this site did not require any observable match to the AAUAAA consensus. It is possible that 3' end formation at this site occurs by a novel mechanism--trans-splicing-dependent cleavage--as deletion of the trans-splice site prevented 3' end formation here. Changing the AGUAAA to AAUAAA also influenced the trans-splicing process: with AGUAAA, most of the gpd-2 product was trans-spliced to SL1, rather than SL2, which is normally used at downstream operon trans-splice sites. However, with AAUAAA, SL2 trans-splicing of gpd-2 was increased. Our results imply that (1) the AAUAAA consensus controls 3' end formation frequency in C. elegans; (2) the AAUAAA is important in determining SL2 trans-splicing events more than 100 bp downstream; and (3) in some circumstances, 3' end formation may occur by a trans-splicing-dependent mechanism.


CHIPing away at the problem of uninsured children.
The State Children's Health Insurance Program (CHIP), enacted one year ago this August, is the largest expansion of health insurance in more than three decades. One of the measures of its success will be whether state officials are able to enroll children who are eligible. Research conducted by Health System Change (HSC) shows that uninsured children are a diverse group, and that for CHIP to be successful, policy makers will need to target programs to specific groups and local market conditions. This Issue Brief discusses why children lack health insurance and the implications for implementing CHIP.


Increased serum levels of soluble CD95 (APO-1/Fas) in relapsing-remitting multiple sclerosis.
CD95/CD95 ligand interactions are critically involved in the negative regulation of peripheral T-cell responses. Here, we report that serum levels of soluble CD95 are significantly elevated in patients with relapsing remitting multiple sclerosis. In a transectional study, CD95 levels did not correlate with clinical disability or lesion formation on magnetic resonance imaging. Longitudinally, Expanded Disability Status Scale changes were associated with high CD95 levels. Interferon-beta (IFNbeta) treatment led to an initial increase and subsequent decline of serum CD95 levels. Interestingly, patients generating neutralizing antibodies to the drug had significantly higher baseline CD95 levels before IFNbeta treatment than those without neutralizing antibodies.


[Inverted papilloma of the bladder. Apropos of 2 cases].
Two cases of transitional cell inverted papilloma are described. The first case was a 44-year-old male patient who presented with hematuria. The second case was an incidental finding during a retropubic adenomectomy procedure. We describe the clinical features of this condition and discuss its histogenesis and treatment. Because of the possibility, although very small, of its evolving to malignancy, these patients must be followed carefully.


Anxiety, depression, and anger in functional gastrointestinal disorders: A Cross-sectional observational study.
Previous studies have identified a link between anger and somatization. However, little is known about the associations between anger and the development and progression of Functional Gastrointestinal Disorders (FGID). The study aim was to determine the associations between FGID and anger, anxiety, and depression. Participants in this cross-sectional observational study were 109 consecutive patients aged 18-64 years with FGID at Gastroenterology Clinic of Başkent University Hospital. A control group comprised of 96 individuals with no chronic gastrointestinal disorders recruited via snowball sampling. Sociodemographic and clinical information were obtained and participants completed the Hospital Anxiety and Depression Scale and the State-Trait Anger Expression Inventory-2. FGID participants scored higher than controls on depression, anxiety, state anger, and anger expression-in. When the FGID group was divided into upper and lower gastrointestinal symptom groups, the lower symptom group showed higher anger expression-out scores than the upper symptom group. Anger may contribute to the etiology and development of FGID. This is the first study to demonstrate a significant psychological difference between individuals with lower and upper FGID. Interdisciplinary collaboration with gastroenterologists and psychiatrists could strengthen FGID evaluation and may improve treatment compliance.


The Madurai Intraocular Lens Study. I: A randomized clinical trial comparing complications and vision outcomes of intracapsular cataract extraction and extracapsular cataract extraction with posterior chamber intraocular lens.
The Madurai Intraocular Lens Study (MIOLS) was designed to compare safety, efficacy, and quality of life outcomes after either intracapsular cataract extraction with aphakic glasses (ICCE-AG) or extracapsular cataract extraction with posterior chamber intraocular lens (ECCE/ PC-IOL). The Madurai Intraocular Lens Study was a nonmasked randomized controlled clinical trial conducted at a single hospital. Thirty-four hundred patients with age-related cataracts and having a best-corrected visual acuity less than or equal to 20/120 in the better eye were randomly assigned to either of the two cataract operative procedures. The main clinical outcomes were safety (complication rates) and efficacy (best-corrected visual acuity at 1 year equal to or better than 20/40). In addition, a subset of 1,700 trial participants received questionnaires before surgery, at 6 months after surgery, and at 1 year after surgery to measure visual functioning and vision-related quality of life. Details of study design, study organization, clinical and quality of life outcome variables, sample size calculations, patient eligibility criteria and recruitment, randomization and masking, participant flow, adherence to follow-up, quality assurance, and statistical methods are presented. The Madurai Intraocular Lens Study has sufficient power to detect clinically significant differences between the treatment options. There were no statistically significant differences between the two treatment groups for any of the major study variables at baseline. A high level of quality assurance was maintained throughout the October 1993 to June 1996 study period. The results should be applicable to all settings where the requisite expertise and resources are present.


Expression of the art/trs protein of HIV and study of its role in viral envelope synthesis.
The art/trs transactivator protein of human immunodeficiency virus (HIV) was expressed in mammalian cells as a 19-kilodalton protein that was immunoreactive with sera from HIV-infected patients. Separate plasmids encoding the art/trs protein, the tat protein, or the envelope glycoprotein gp120 were used to demonstrate that both art/trs and tat are absolutely required for the synthesis of gp120 from its cognate messenger RNA. In addition, both the tat and art/trs proteins influence the level of envelope RNA. The results suggest that art/trs and tat may be ideal targets for potential anti-HIV agents in AIDS therapy.


Phosphopeptide-selective column-switching RP-HPLC with a titania precolumn.
A methodology of phosphopeptide-selective analysis coupled with column-switching HPLC utilizing titania as precolumn media is presented. Phosphopeptides were selectively enriched on titania packing within a protein/peptide mixture without any additional procedure, and analyzed by column-switching high-performance liquid chromatography. First, phospho-compounds were separated from complex mixtures by trapping them under acidic conditions on a titania packing, where non-phosphorylated compounds were effused out of the precolumn. Subsequently, phospho-compounds were desorbed from the titania column under a specific condition and analyzed. The behavior of phospho-compounds on a titania surface, especially adsorption/desorption, was precisely examined and optimized. A phosphoric buffer was successively employed for the elution of phosphopeptides on a titania surface by competition with the free phosphate group. From the successes of a selective concentration/analysis of phosphopeptides with column-switching HPLC with a titania precolumn, a novel phosphopeptide-selective RP-HPLC analysis has been shown to have an application possibility as a tool for phosphoproteomics.


Relationship Between Atypical Sexual Fantasies, Behavior, and Pornography Consumption.
Paraphilia is a condition in which the sexual excitement relies on fantasizing and/or participating in unusual sexual behaviors although the line between "normal" and "abnormal" has been disputed. The project aimed to explore which sexual fantasies and behaviors are common and uncommon in the general population. Furthermore, the relationship between sexual fantasies, sexual behaviors, and problematic pornography consumption was examined. Finally, the impact of gender was assessed. Analyses were conducted on a sample of 139 participants. Correlations were found between fantasies, behaviors, and problematic pornography consumption. Furthermore, gender differences were found for both sexual fantasies and problematic pornography consumption. Finally, multiple regression revealed that age, gender [Men], fantasies, and behaviors were significant predictor of problematic pornography consumption. Those findings are in-line with previous studies which highlighted that the threshold to consider a sexual practice as being abnormal needs to be reconsidered on the basis of self-reported fantasies and behaviors in the general population.