Biventricular thrombus in hypereosinophilic syndrome presenting with shortness of breath.
A 48 years old male presented to clinic with 12 months of low grade fever with shortness of breath which has progressively worsened with no associated weight loss, night sweats or loss of appetite. There was no prior history of chronic illness before the current illness. Laboratory workup revealed a high white blood cell count with predominant eosinophils. Chest X-ray was normal. Transthoracic echocardiography and Cardiac Magnetic Resonance showed biventricular thrombi. On further extensive workup the findings were consistent with hypereosinophilic syndrome. The patient was started on oral steroids, hydroxyurea, imatanib mesylate and oral anticoagulation. The patient responded to the treatment with complete resolution of his symptoms over the course of few months. The repeat Echocardiogram after a year showed normal left ventricular systolic and diastolic function with complete resolution of biventricular thrombi.


Older Australians Can Achieve High Adherence to the Mediterranean Diet during a 6 Month Randomised Intervention; Results from the Medley Study.
Adherence to a Mediterranean diet (MedDiet) is thought to be achievable in non-Mediterranean regions, but this has yet to be investigated. We aimed to determine if an older Australian population could adhere to a MedDiet for six months. We conducted a randomised, parallel dietary intervention trial with two dietary arms: the Mediterranean diet (MedDiet) group and the habitual diet (HabDiet) control group. A 15-point Mediterranean diet adherence score and food and nutrient intakes were estimated from three-day weighed food records collected at baseline, two and four months. Erythrocyte fatty acids, serum carotenoids and urinary metabolites were assessed at baseline, three and six months. We enrolled 166 participants; 152 commenced and 137 completed the study (70 in the MedDiet group, 67 in the HabDiet group). Adherence scores were significantly higher in the MedDiet group at two months (between group difference 2.2, 95% CI 1.3, 2.9) and four months (between group difference 2.6, 95% CI 1.9, 3.3). Consumption of vegetables, fruits, fish, legumes, nuts and olive oil significantly increased in the MedDiet group compared to the control, and discretionary food intake decreased (p < 0.01). Measures of compliance including serum β-carotene, lycopene and erythrocyte monounsaturated fatty acids were significantly higher in the MedDiet group at three and six months (p < 0.05). Our results indicate that a population of older Australians can adopt a Mediterranean diet over a six month period.


Percutaneous Kyphoplasty: Is Bilateral Approach Necessary?
A multicenter retrospective study of patients who underwent unilateral and bilateral balloon kyphoplasty. The aim of this study was to compare the radiographic and clinical results of unilateral and bilateral balloon kyphoplasty to treat osteoporotic vertebral compression fractures. Percutaneous kyphoplasty has long been used as a successful method in the treatment of osteoporotic vertebral compression fractures. Although the bilateral approach is considered to be the mainstay application of percutaneous kyphoplasty, the unilateral approach has also been shown to be sufficient and even more effective in some cases. A total of 87 patients who underwent percutaneous kyphoplasty due to osteoporotic vertebral compression fractures between 2009 and 2016 were retrospectively evaluated and divided into two groups as patients who underwent unilateral or bilateral percutaneous kyphoplasty. Unilateral percutaneous kyphoplasty was performed in 36 and bilateral percutaneous kyphoplasty in 51 patients. The groups were compared in terms of clinical outcomes, radiological findings, and complications. Clinical outcomes were evaluated using Visual Analogue Scale and Oswestry Disability Index and the radiological findings were evaluated by comparing the preoperative and postoperative day 1 and year 1 values of anterior, middle, and posterior vertebral heights and kyphosis angle. Clinical improvement occurred in both groups but no significant difference was observed. In radiological workup, no significant difference was found between the groups in terms of improvements in vertebral heights and kyphosis angle. Operative time and the amount of cement used for the surgery were significantly lower in the patients that underwent unilateral kyphoplasty. Unilateral percutaneous kyphoplasty is as effective as bilateral percutaneous kyphoplasty both radiologically and clinically. Operative time and the amount of cement used for the surgery are significantly lower in unilateral kyphoplasty, which may play a role in decreasing complication rates. 3.


Expression of xanthine oxidoreductase in mouse mammary epithelium during pregnancy and lactation: regulation of gene expression by glucocorticoids and prolactin.
In the mammary gland of virgin mice, xanthine oxidoreductase (XOR) enzymic activity is barely measurable. a high increase in the levels of the enzyme is observed during the last days of pregnancy and during lactation, and this is parallelled by an elevation in the amounts of the respective protein and transcript. In situ hybridization experiments demonstrate that the XOR mRNA is specifically expressed in the alveolar epithelial cells of the mammary gland. In HC11 cells, a model culture system for normal breast epithelium, the levels of XOR enzymic activity are dose- and time-dependently induced by dexamethasone, and a further synergistic augmentation is observed in the presence of dexamethasone plus prolactin. Increased XOR gene expression is consequent on glucocorticoid receptor activation, as indicated by sensitivity to the specific receptor antagonist RU486. In addition, the phenomenon is likely to involve protein phosphorylation and dephosphorylation events, as suggested by modulation of XOR mRNA by tyrosine kinase and phosphatase inhibitors.


Using the IAT to predict ethnic and racial discrimination: small effect sizes of unknown societal significance.
Greenwald, Banaji, and Nosek (2015) present a reanalysis of the meta-analysis by Oswald, Mitchell, Blanton, Jaccard, and Tetlock (2013) that examined the effect sizes of Implicit Association Tests (IATs) designed to predict racial and ethnic discrimination. We discuss points of agreement and disagreement with respect to methods used to synthesize the IAT studies, and we correct an error by Greenwald et al. that obscures a key contribution of our meta-analysis. In the end, all of the meta-analyses converge on the conclusion that, across diverse methods of coding and analyzing the data, IAT scores are not good predictors of ethnic or racial discrimination, and explain, at most, small fractions of the variance in discriminatory behavior in controlled laboratory settings. The thought experiments presented by Greenwald et al. go well beyond the lab to claim systematic IAT effects in noisy real-world settings, but these hypothetical exercises depend crucially on untested and, arguably, untenable assumptions.


Pressure-induced superconductivity in CaC2.
Carbon can exist as isolated dumbbell, 1D chain, 2D plane, and 3D network in carbon solids or carbon-based compounds, which attributes to its rich chemical binding way, including sp-, sp(2)-, and sp(3)-hybridized bonds. sp(2)-hybridizing carbon always captures special attention due to its unique physical and chemical property. Here, using an evolutionary algorithm in conjunction with ab initio method, we found that, under compression, dumbbell carbon in CaC2 can be polymerized first into 1D chain and then into ribbon and further into 2D graphite sheet at higher pressure. The C2/m structure transforms into an orthorhombic Cmcm phase at 0.5 GPa, followed by another orthorhombic Immm phase, which is stabilized in a wide pressure range of 15.2-105.8 GPa and then forced into MgB2-type phase with wide range stability up to at least 1 TPa. Strong electron-phonon coupling λ in compressed CaC2 is found, in particular for Immm phase, which has the highest λ value (0.562-0.564) among them, leading to its high superconducting critical temperature Tc (7.9∼9.8 K), which is comparable with the 11.5 K value of CaC6. Our results show that calcium not only can stabilize carbon sp(2) hybridization at a larger range of pressure but also can contribute in superconducting behavior, which would further ignite experimental and theoretical interest in alkaline-earth metal carbides to uncover their peculiar physical properties under extreme conditions.


Synergistic soft tissue infections of the perineum.
Seven patients with necrotizing soft tissue infections of the perineum are described. Predisposing factors related to infection were present in four patients (diabetes mellitus, multiple myeloma, HIV, and a poorly defined immunodeficiency syndrome). Anaerobic and facultative anaerobic bacteria were cultured in each case. Two patients required skin graft closure of the debrided wounds, with the remaining wounds closed by contracture and epithelialization. A diverting sigmoid colostomy to facilitate wound care was performed on one patient who had complete dissolution of all anal sphincters. The role of hyperbaric oxygen therapy in four patients was of uncertain value.


ACR Appropriateness Criteria® Soft-Tissue Masses.
Imaging is an integral component of the evaluation of patients with a suspected soft-tissue mass. Imaging can not only confirm the presence of a mass but can provide essential information necessary for diagnosis, local staging, and biopsy planning. Although the objectives of the evaluation have not changed, the choices available for imaging of musculoskeletal masses have evolved dramatically in recent years. The purpose of this document is to identify the most common clinical scenarios and the most appropriate imaging for their assessment on the basis of the current literature and to provide general guidance for those scenarios that are not specifically addressed. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision include an extensive analysis of current medical literature from peer reviewed journals and the application of well-established methodologies (RAND/UCLA Appropriateness Method and Grading of Recommendations Assessment, Development, and Evaluation or GRADE) to rate the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where evidence is lacking or equivocal, expert opinion may supplement the available evidence to recommend imaging or treatment.


Effects of dose, administration route, and/or vehicle on decabromodiphenyl ether concentrations in plasma of maternal, fetal, and neonatal rats and in milk of maternal rats.
The effects of route and vehicle on blood and milk levels of decabromodiphenyl ether (DecaBDE; CASRN 1163-19-5) were investigated in the rat to assist in the design and conduct of a developmental neurotoxicity study. Blood plasma and/or milk concentrations were determined in dams, fetuses, and/or nursing pups after repeated DecaBDE administration by gavage throughout gestation or gestation and lactation using corn oil (CO) or soyaphospholipon/Lutrol F 127-water (SPL) as the vehicle. The impact of vehicle on plasma levels was also investigated in pups derived from naive dams after a single postnatal dose. This study reports for the first time fetal and neonatal plasma concentrations concurrent with those of maternal plasma and/or milk. Higher concentrations of DecaBDE were achieved in plasma and in milk with CO than with SPL. Furthermore, pups derived from dams treated with only SPL were lower in body weight, compared with those from dams treated with either CO, CO and DecaBDE, or SPL and DecaBDE. The study further shows that exposure to DecaBDE is relatively consistent across the dose range of 100 to 1000 mg/(kg · day) when administered in CO.


Recurrent Glioblastoma Treated with Recombinant Poliovirus.
The prognosis of patients with recurrent World Health Organization (WHO) grade IV malignant glioma is dismal, and there is currently no effective therapy. We conducted a dose-finding and toxicity study in this population of patients, evaluating convection-enhanced, intratumoral delivery of the recombinant nonpathogenic polio-rhinovirus chimera (PVSRIPO). PVSRIPO recognizes the poliovirus receptor CD155, which is widely expressed in neoplastic cells of solid tumors and in major components of the tumor microenvironment. We enrolled consecutive adult patients who had recurrent supratentorial WHO grade IV malignant glioma, confirmed on histopathological testing, with measurable disease (contrast-enhancing tumor of ≥1 cm and ≤5.5 cm in the greatest dimension). The study evaluated seven doses, ranging between 107 and 1010 50% tissue-culture infectious doses (TCID50), first in a dose-escalation phase and then in a dose-expansion phase. From May 2012 through May 2017, a total of 61 patients were enrolled and received a dose of PVSRIPO. Dose level -1 (5.0×107 TCID50) was identified as the phase 2 dose. One dose-limiting toxic effect was observed; a patient in whom dose level 5 (1010 TCID50) was administered had a grade 4 intracranial hemorrhage immediately after the catheter was removed. To mitigate locoregional inflammation of the infused tumor with prolonged glucocorticoid use, dose level 5 was deescalated to reach the phase 2 dose. In the dose-expansion phase, 19% of the patients had a PVSRIPO-related adverse event of grade 3 or higher. Overall survival among the patients who received PVSRIPO reached a plateau of 21% (95% confidence interval, 11 to 33) at 24 months that was sustained at 36 months. Intratumoral infusion of PVSRIPO in patients with recurrent WHO grade IV malignant glioma confirmed the absence of neurovirulent potential. The survival rate among patients who received PVSRIPO immunotherapy was higher at 24 and 36 months than the rate among historical controls. (Funded by the Brain Tumor Research Charity and others; ClinicalTrials.gov number, NCT01491893 .).


Nasal carriage of methicillin-resistant Staphylococcus aureus among pediatricians in Taiwan.
Health care workers (HCWs) are at the interface between hospitals and communities. The survey for methicillin-resistant Staphylococcus aureus (MRSA) carriage among HCWs has mostly been conducted to investigate outbreaks or endemics. Community-associated MRSA are prevalent among children in Taiwan. We conducted this study to better understand the carriage rate of MRSA among pediatricians in non-outbreak situations in Taiwan,. A total of 220 pediatricians from Taiwan who attended the annual meeting of Taiwan Pediatric Association in April, 2010 were recruited to participate in this study and were sampled from the nares for the detection of MRSA by polymerase chain reaction (PCR) and further by culture. The following molecular analyses were performed, including pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), typing of staphylococcal cassette chromosome mec (SCCmec) and the presence of Panton-Valentine leukocidin (PVL) genes. MRSA was detected from 15 attendees (6.8%) by PCR. MRSA-colonized attendees had a significantly lower rate (0.041) of working in the medical center, while borderline significantly higher rate of working in the Regional Hospital (p=0.056), than those without MRSA colonization. From those 15 samples, 12 MRSA isolates were identified by culture and molecularly characterized. Three PFGE patterns, two sequence types (ST 59, ST 508), and two SCCmec types (IV and VT) were identified, respectively. Five isolates, including three carrying SCCmec types VT, were PVL-positive. All 12 isolates were susceptible to vancomycin, teicoplanin, linezolid, fusidic acid, trimethoprim/sulfamethoxazole, and doxycyclin, and resistant to penicillin. Around seven percent of pediatricians in Taiwan harbored CA-MRSA in their nares.


Characterisation of an amphioxus Fringe gene and the evolution of the vertebrate segmentation clock.
In mouse and chick embryos, cyclic expression of lunatic fringe has an important role in the regulation of mesoderm segmentation. We have isolated a Fringe gene from the protochordate amphioxus. Amphioxus is the closest living relative of the vertebrates, and has mesoderm that is definitively segmented in a manner that is similar to, and probably homologous with, that of vertebrates. AmphiFringe is placed basal to vertebrate Fringe genes in molecular phylogenetic analyses, indicating that the duplications that formed radical-, manic- and lunatic fringe are specific to the vertebrate lineage. AmphiFringe expression was detected in the anterior neural plate of early neurulae, where it resolved into a series of segmental patches by the mid-neurulae stage. No AmphiFringe transcripts were detected in the mesoderm. Based on these observations, we propose a model depicting a successive recruitment of Fringe in the maintenance then regulation of segmentation during vertebrate evolution.


The cAMP-regulated enhancer-binding protein ATF-1 activates transcription in response to cAMP-dependent protein kinase A.
Many promoters respond transcriptionally to elevated levels of cAMP through the cAMP-responsive enhancer (CRE). Several proteins have been characterized which bind to the CRE and presumably modulate CRE-dependent transcription. Of these CRE-binding proteins, only CREB has been shown to be activated by cAMP-dependent protein kinase A (PKA), and as such, CREB represents the only basis for our understanding of cAMP-regulated transcriptional activity. In this report, we describe the complete cDNA sequence of another CRE-binding protein, ATF-1. This protein contains a consensus phosphorylation site for PKA and shares extensive homology with CREB in the region surrounding and carboxyl-terminal to the PKA site. ATF-1 does not contain sequences homologous to the glutamine-rich amino-terminal domain found in CREB, however. ATF-1, like CREB, is expressed in a wide variety of cell types, and ATF-1 is capable of dimerizing with CREB. Both ATF-1 homodimers and ATF-1/CREB heterodimers bind to the CRE but not to the related phorbol ester response element. ATF-1 is as active as CREB in its ability to mediate the transcriptional effects of PKA, and, because ATF-1 has a smaller effect on basal expression, it is actually more responsive than CREB to cAMP. These findings indicate that CREB is not unique in its ability to mediate cAMP-dependent transcriptional regulation.


"Memory bytes" - molecular match for CaMKII phosphorylation encoding of microtubule lattices.
Learning, memory and long-term potentiation (LTP) are supported by factors including post-synaptic calcium ion flux activating and transforming the hexagonal calcium-calmodulin kinase II (CaMKII) holoenzyme. Upon calcium-induced activation, up to six kinase domains extend upward, and up to six kinase domains extend downward from the CaMKII association domain, the fully activated holoenzyme resembling a robotic insect 20 nanometers in length. Each extended kinase domain can be phosphorylated, and able to phosphorylate other proteins, thus potentially further encoding synaptic information at intraneuronal molecular sites for memory storage, processing and distribution. Candidate sites for phosphorylation-encoded molecular memory include microtubules, cylindrical lattice polymers of the protein tubulin. Using molecular modeling, we find spatial dimensions and geometry of the six extended CaMKII kinase domains can precisely match those of microtubule hexagonal lattice neighborhoods (both A- and B-lattices), and show two feasible phosphorylation mechanisms. In one, phosphorylation sites (e.g., valine 208) on a CaMKII extended kinase domain interact with serine 444 on a C-terminal "tail" of tubulin. In the second, the CaMKII kinase domain unfurls, enabling phosphorylation sites to contact threonine and serine sites on the tubulin surface. We suggest sets of six CaMKII kinase domains phosphorylate hexagonal microtubule lattice neighborhoods collectively, e.g., conveying synaptic information as ordered arrays of six "bits", and thus a "byte", with (minimally) 2⁶ (64) possible bit states per CaMKII-microtubule interaction. We model two levels of interaction between CaMKII and microtubules, suggesting a testable framework for molecular memory encoding.


Treatment of scleroderma.
The treatment of systemic sclerosis (scleroderma) is difficult and remains a great challenge to the clinician. Because the cause is unknown, therapies are directed to improve peripheral blood circulation with vasodilators and antiplatelet aggregation drugs, to prevent the synthesis and release of harmful cytokines with immunosuppressant drugs, and to inhibit or reduce fibrosis with agents that reduce collagen synthesis or enhance collagenase production. The purpose of this review is to critically analyze conventional and new treatments of systemic sclerosis and localized scleroderma. The therapeutic options discussed for the treatment of systemic sclerosis include the use of (1) vasodilators (calcium channel blockers [nifedipine], angiotensin-converting enzyme inhibitors [captopril, losartan potassium], and prostaglandins [iloprost, epoprostenol]), (2) immunosuppressant drugs (methotrexate, cyclosporine, cyclophosphamide, and extracorporeal photopheresis), and (3) antifibrotic agents (D-penicillamine, colchicine, interferon gamma, and relaxin). The treatment options reviewed for localized scleroderma include the use of corticosteroids, vitamin D analogues (calcitriol, calcipotriene), UV-A, and methotrexate. Preliminary reports on new therapies for systemic sclerosis are also considered. These include the use of minocycline, psoralen-UV-A, lung transplantation, autologous stem cell transplantation, etanercept, and thalidomide.


Societal costs and burden of hereditary transthyretin amyloidosis polyneuropathy.
Background: Hereditary transthyretin amyloidosis polyneuropathy (ATTRv-PN) is a rare life-threatening disease that imposes considerable mortality and morbidity associated with increased costs, high social support and productivity losses. This study aims to estimate the societal costs and burden of ATTRv-PN.Methods: A cost-of-illness (COI) and burden of disease model were specified from a societal perspective, using a prevalence-based approach. Direct and indirect costs were included. Healthcare resource use was retrieved from public databases, previous Portuguese studies and the literature. The burden of disease was expressed in terms of disability-adjusted life years (DALYs), as defined by the World Health Organisation.Results: In 2016, the total annual COI of ATTRv-PN in Portugal was 52,502,796€ and the mean cost per patient was 28,152€ (79% direct; 21% indirect costs). Treatments accounted for 52% of total costs, while 0.18% were devoted to disease prevention. A total of 2056 DALYs were lost, 26% due to disability and 74% due to death.Conclusions: Annual costs and burden of ATTRv-PN were considerable but within the range of other rare diseases. Policies and public interventions to prevent and reduce the burden of disease should be prioritised, since patients experience excess morbidity, mortality and total costs will likely increase in the future.


Image-guided pleural biopsy.
Pleural diseases are a common and increasing clinical problem. Establishing accurate diagnosis is an essential step in management of these patients, and approximately 40% of pleural effusions will remain undiagnosed after initial diagnostic thoracocentesis. Obtaining pleural tissue (by blind, image-guided or thoracoscopic pleural biopsy) is therefore a key procedure. Recent evidence provides important information on the relative value of each of these techniques. For the diagnosis of malignant pleural disease, both thoracoscopic and image-guided biopsy have a far higher diagnostic yield than blind pleural biopsy. Cutting needle biopsies have a higher diagnostic yield in malignancy (and especially mesothelioma) compared with fine needle aspiration. The complication rate of image-guided biopsy is low. Rates of biopsy site tract invasion by mesothelioma may be lower using smaller biopsy ports, as used for image-guided pleural biopsy. Blind pleural biopsy should no longer be conducted for the study of malignant pleural disease if facilities for other techniques are available. Image-guided and thoracoscopic biopsies have similarly high diagnostic rates, and are complementary techniques used in different clinical situations. Further studies assessing biopsy tract site invasion from mesothelioma with different biopsy techniques are required.


Albino mice as an animal model for infantile nystagmus syndrome.
Individuals with oculocutaneous albinism are predisposed to visual system abnormalities affecting the retina and retinofugal projections, which may lead to reduced visual acuity and Infantile Nystagmus Syndrome (INS). Due to absence of an established mammalian animal model, mechanisms underlying INS remain elusive. In this study, we screened wild-type mice of varying pigmentation for ocular motor abnormalities in order to identify a possible mouse model for INS. Three albino mouse strains (CD1, BALB/c, DBA/1), and two normally pigmented strains (129S6, C57BL/6) were screened using infrared oculography. Varying visual stimuli (black or white background, stationary pattern, optokinetic, i.e., horizontally rotating pattern) were displayed to the full (fVF) or anterior visual field (aVF) of the restrained mouse. We found spontaneous nystagmus, specifically jerks and oscillations, in albino mice under all experimental conditions. Median eye velocity was between 0.8 and 3.4 deg/s, depending on the strain. In contrast, the eyes in pigmented mice were nearly stable with a median absolute eye velocity of below 0.4 deg/s. In albino mice, fVF optokinetic stimuli elicited an optokinetic response (OKR) in the correct direction, albeit with superimposed oscillations. However, aVF optokinetic stimuli evoked reversed OKR in these strains, a well known feature of INS. Based on our results, we endorse the investigated albino mouse strains as new animal models for INS.


An update on DNA-based BRCA1/BRCA2 genetic counseling in hereditary breast cancer.
The identification of BRCA1 and BRCA2 mutations has enabled physicians to identify persons at high risk for carcinoma of the breast and ovary in hereditary breast-ovarian cancer (HBOC) families. Many physicians have limited knowledge about the effective translation of these new discoveries into clinical practice settings. This problem is further confounded by the limited number of genetic counselors who have experience with cancer genetics. Genetic counseling about DNA test results was provided to 420 patients from 37 HBC/HBOC families. Descriptive data were collected and recorded about their responses to questions posed immediately before and after test results were disclosed. Findings disclosed a significant tendency of patients to overestimate rather than underestimate their risk (P < .001) prior to receiving results. The chief reason for declining to receive results was fear of insurance discrimination. The primary reason that patients sought test results was for their children. Most women reported that, if testing identified them as mutation carriers, they would consider lifetime surveillance and prophylactic surgery. Responses to DNA test results were varied and often unpredictable. Counseling by an appropriately educated and skilled professional is essential to assist people in making decisions regarding testing and health management.


Rapid emergence of a ceftazidime-resistant Burkholderia multivorans strain in a cystic fibrosis patient.
Burkholderia multivorans poses a serious health threat to cystic fibrosis patients due to innate resistance to multiple antibiotics and acquisition of resistance to a range of antibiotics due to the frequent use of antibiotics to treat chronic infections. Monitoring antibiotic susceptibility is crucial to managing patient care. We identified the rapid emergence of a ceftazidime-resistant strain in a single patient within four days during a hospitalization for treatment of an exacerbation. B. multivorans was isolated from expectorated sputum samples using Burkholderia cepacia selective agar. A macrodilution assay was performed on all isolates to determine the minimum inhibitory concentration of ceftazidime. Approximately 4000 colonies were scored to identify the percent of ceftazidime-resistant colonies. Extracted DNA was used to determine the total bacterial counts and abundance of B. multivorans using quantitative PCR. An increase from no detectable B. multivorans ceftazidime-resistant colonies to over 75% of all colonies tested occurred within a four-day period. The resistant population remained dominant in 6 of the 8 samples in the following 17 months of the study. qPCR revealed an association between change in the percent of resistant colonies and abundance of B. multivorans, but not of total bacteria. No association was found between the acquisition of resistance to ceftazidime and other antibiotics commonly used to treat B. multivorans infections. The rapid emergence of a ceftazidime-resistant by B. multivorans strain occurred during a hospitalization while under selective pressure of antibiotics. The resistant strain maintained dominance in the B. multivorans population which resulted in an overall decline in a patient health and treatment efficacy.


Changes in Adrenal Androgens and Steroidogenic Enzyme Activities From Ages 2, 4, to 6 Years: A Prospective Cohort Study.
The levels of adrenal androgens are increased through the action of steroidogenic enzymes with morphological changes in the adrenal zona reticularis. We investigated longitudinal changes in androgen levels and steroidogenic enzyme activities during early childhood. From a prospective children's cohort, the Environment and Development of Children cohort, 114 boys and 86 girls with available blood samples from ages 2, 4, and 6 years were included. Serum concentrations of adrenal androgens using liquid chromatography-tandem mass spectrometry and steroidogenic enzyme activity calculated by the precursor/product ratio. During ages 2 to 4 years, 17,20-lyase and dehydroepiandrosterone (DHEA) sulfotransferase activities increased (P < 0.01 for both in boys). During ages 4 to 6 years, 17,20-lyase activity persistently increased, but 3β-hydroxysteroid dehydrogenase (HSD) and 17β-HSD activities decreased (P < 0.01 for all). Serum DHEA sulfate (DHEA-S) levels persistently increased from 2, 4, to 6 years, and DHEA, 17-hydroxyprogesterone, and androstenedione levels increased during ages 4 to 6 years (P < 0.01 for all). Serum DHEA-S levels during early childhood were associated with body mass index z-scores (P = 0.001 in only boys). This study supports in vivo human evidence of increased 17,20-lyase and DHEA sulfotransferase activities and decreased 3β-HSD activity during early childhood.


Do User-Applied Safety Labels on Medication Syringes Reduce the Incidence of Medication Errors During Rapid Medical Response Intervention for Deteriorating Patients in Wards? A Systematic Search and Review.
Intravenous medication errors (MEs) occur during medical emergency situations. An initiative, not yet in common practice, that could address these errors is safety labeling. The aim of this review was to identify and appraise research evidence related to the impact of user-applied medication safety labeling on reducing the incidence of MEs during rapid medical response intervention for patient deterioration in the ward setting. A systematic search and review framework was used to conduct the review. A comprehensive database search was conducted of BioMed Central, Clinical Trials, Cumulative Index to Nursing and Allied Health Literature, Expanded Academic ASAP, Joanna Briggs Institute, MEDLINE, OVID, ProQuest Central, PubMed, Wiley Online Library, and World Health Organization Library. The Young and Solomon (2009) critical appraisal tool was used to critically appraise the identified research articles. Each article was then analyzed using a thematic network strategy to identify commonality. Four primary themes were identified; they were as follows: MEs occur during medical emergency responses (MERs); MEs occur throughout the medication administration process; MERs are stressful and are associated with MEs; and role of medication labeling in reducing MEs during MERs. Greater vigilance is required by health professionals during the medication administration process. The implementation of specific medication safety labeling into the MER could be beneficial in reducing the overall incidence of MEs. Further research is required to validate the merits of a MER medication safety labeling system.


In vivo efficacy of human immunodeficiency virus neutralizing antibodies: estimates for protective titers.
The definition of plasma neutralizing antibody titers capable of controlling human immunodeficiency virus (HIV) infection in vivo is considered a critical step in vaccine development. Here we provide estimates for effective neutralization titers by assessing samples from a recent passive immunization trial with the neutralizing monoclonal antibodies (MAbs) 2G12, 2F5, and 4E10 using an analytic strategy that dissects the contributions of these MAbs to the total neutralization activity in patient plasma. Assessment of neutralization activities for six responding patients with partial or complete control of viremia during the MAb treatment and for the eight nonresponding patients revealed a significant difference between these groups: Among responders, MAb-mediated activity exceeded the autologous neutralization response by 1 to 2 log units (median difference, 43.3-fold), while in the nonresponder group, the autologous activity prevailed (median difference, 0.63-fold). In order to reach a 50% proportion of the responders in our study cohort, MAb neutralizing titers higher than 1:200 were required based on this analysis. The disease stage appears to have a significant impact on the quantities needed, since titers above 1:1,000 were needed to reach the same effect in chronic infection. Although our analysis is based on very small sample numbers and thus cannot be conclusive, our data provide a first estimate on how in vitro-measured neutralizing antibody activity can relate to in vivo efficacy in controlling HIV infection and may therefore provide valuable information for vaccine development. Interestingly, lower neutralizing antibody levels showed an effect in acute compared to chronic infection, suggesting that in early disease stages, therapeutic vaccination may show promise. Equally, this raises hopes that a preventive vaccine could become effective at comparatively lower neutralizing antibody titers.


Understanding surfactant/graphene interactions using a graphene field effect transistor: relating molecular structure to hysteresis and carrier mobility.
Manipulation of transport hysteresis on graphene transistors and understanding electron transfer between graphene and polar/ionic adsorbates are important for the development of graphene-based sensor devices and nonvolatile memory electronics. We have investigated the effects of commonly used surfactants for graphene dispersion in aqueous solution on transport characteristics of graphene transistors. The adsorbates are found to transfer electrons to graphene, scatter carrier transport, and induce additional electron-hole puddles when the graphene is on an SiO(2) substrate. We relate the change in transport characteristics to specific chemical properties of a series of anionic, cationic, and neutral surfactants using a modification of a self-consistent transport theory developed for graphene. To understand the effects of surfactant adsorbates trapped on either side of the graphene, suspended devices were fabricated. Strong hysteresis is observed only when both sides of the graphene were exposed to the surfactants, attributable to their function as charge traps. This work is the first to demonstrate the control of hysteresis, allowing us to eliminate it for sensor and device applications or to enhance it to potentially enable nonvolatile memory applications.


Correlates of household smoking bans among Chinese Americans.
No population-based data are available on the degree to which Chinese Americans have adopted smoke-free household policies and whether these policies are effective in reducing environmental tobacco smoke (ETS) exposure. The present study examines the prevalence of smoke-free home rules among Chinese Americans living in New York City, describes predictors of adopting full smoking bans in the home, and explores the association between household smoking restrictions and ETS exposure at home. In-person interviews using a comprehensive household-based survey were conducted with 2,537 adults aged 18-74 years. Interviews were conducted in Mandarin, Cantonese, and other Chinese dialects. A total of 66% of respondents reported that smoking was not allowed inside the home, 22% reported a partial ban on smoking in the home, and 12% reported no smoking ban. Among current smokers, 38% reported a full household smoking ban. Current smoking status was the strongest predictor of less restrictive household smoking policies. Knowledge of the dangers of ETS, support of smoke-free air legislation, years in the United States, gender, income, and marital status also were associated with household smoking bans. Those living with a total household smoking ban were significantly less likely to report 30-day exposure to ETS than were those living in homes with a partial ban or no ban (7% vs. 68% and 73%, respectively). In homes of smokers and nonsmokers alike, exposure to ETS remains high. Smoke-free home rules and interventions among smokers and nonsmokers to raise awareness of the dangers of ETS have the potential to significantly reduce exposure to household ETS among this immigrant population.


Predictive validity of the pain and impairment relationship scale in a chronic nonmalignant pain population.
To examine the predictive and incremental validity of the Pain and Impairment Relationship Scale (PAIRS) and to determine its ability to measure changes in pain beliefs following interdisciplinary treatment. A before-after treatment design. A comprehensive interdisciplinary pain center at a large midwest university medical center. A cognitive-behavioral approach to pain management. The day-long program lasted 5 days a week for 4 weeks. Interventions were designed to improve physical functioning, reduce use of health care, and improve pain coping. Six-month follow-up outcome measures included interference with daily activity, pain severity, and life control as measured by the Multidimensional Pain Inventory, medication use measured by the Medication Quantification Scale, depression measured by the Beck Depression Inventory, and the number of health care visits and pain-related hospitalizations. Pretreatment PAIRS scores correlated significantly with interference with daily activities, pain severity, life control, health care visits, and depression. Stronger correlations were obtained between posttreatment PAIRS scores and all follow-up outcome measures. Posttreatment PAIRS scores accounted for a significant portion of the variance beyond that in demographic variables and pretreatment PAIRS scores in all but one of the follow-up measures. PAIRS scores changed significantly (p < .0001 ) in a positive direction after treatment. The PAIRS has excellent predictive validity, and can be used effectively to monitor individual and programmatic changes.


Sexual health (excluding reproductive health, intimate partner violence and gender-based violence) and COVID-19: a scoping review.
The COVID-19 pandemic has exposed and exacerbated existing socioeconomic and health disparities, including disparities in sexual health and well-being. While there have been several reviews published on COVID-19 and population health disparities generally-including some with attention to HIV-none has focused on sexual health (ie, STI care, female sexual health, sexual behaviour). We have conducted a scoping review focused on sexual health (excluding reproductive health (RH), intimate partner violence (IPV) and gender-based violence (GBV)) in the COVID-19 era, examining sexual behaviours and sexual health outcomes. A scoping review, compiling both peer-reviewed and grey literature, focused on sexual health (excluding RH, IPV and GBV) and COVID-19 was conducted on 15 September 2020. Multiple bibliographical databases were searched. Study selection conformed to Joanna Briggs Institute (JBI) Reviewers' Manual 2015 Methodology for JBI Scoping Reviews. We only included English-language original studies. We found that men who have sex with men may be moving back toward pre-pandemic levels of sexual activity, and that STI and HIV testing rates seem to have decreased. There was minimal focus on outcomes such as the economic impact on sexual health (excluding RH, IPV and GBV) and STI care, especially STI care of marginalised populations. In terms of population groups, there was limited focus on sex workers or on women, especially women's sexual behaviour and mental health. We noticed limited use of qualitative techniques. Very few studies were in low/middle-income countries (LMICs). Sexual health research is critical during a global infectious disease pandemic and our review of studies suggested notable research gaps. Researchers can focus efforts on LMICs and under-researched topics within sexual health and explore the use of qualitative techniques and interventions where appropriate.


Addition of inhaled treprostinil to oral therapy for pulmonary arterial hypertension: a randomized controlled clinical trial.
This study assessed the efficacy and safety of inhaled treprostinil in pulmonary arterial hypertension (PAH) patients receiving therapy with either bosentan or sildenafil. There is no cure for PAH, despite effective treatments, and outcomes remain suboptimal. The addition of inhaled treprostinil, a long-acting prostacyclin analog, might be a safe and effective treatment addition to other PAH-specific oral therapies. Two hundred thirty-five PAH patients with New York Heart Association (NYHA) functional class III (98%) or IV symptoms and a 6-min walk distance (6MWD) of 200 to 450 m while treated with bosentan (70%) or sildenafil were randomized to inhaled treprostinil (up to 54 mug) or inhaled placebo 4 times daily. The primary end point was peak 6MWD at 12 weeks. Secondary end points included time to clinical worsening, Borg Dyspnea Score, NYHA functional class, 12-week trough 6MWD, 6-week peak 6MWD, quality of life, and PAH signs and symptoms. The biomarker N-terminal pro-brain natriuretic peptide (NT-proBNP) was assessed. Twenty-three patients withdrew from the study prematurely (13 treprostinil, 10 placebo). The Hodges-Lehmann between-treatment median difference in change from baseline in peak 6MWD was 19 m at week 6 (p = 0.0001) and 20 m at week 12 (p = 0.0004). Hodges-Lehmann between-treatment median difference in change from baseline in trough 6MWD at week 12 was 14 m (p = 0.0066). Quality of life measures and NT-proBNP improved on active therapy. There were no improvements in other secondary end points, including time to clinical worsening, Borg Dyspnea Score, NYHA functional class, and PAH signs and symptoms. Inhaled treprostinil was safe and well-tolerated. This trial demonstrates that, among PAH patients who remain symptomatic on bosentan or sildenafil, inhaled treprostinil improves exercise capacity and quality of life and is safe and well-tolerated. (TRIUMPH I: Double Blind Placebo Controlled Clinical Investigation Into the Efficacy and Tolerability of Inhaled Treprostinil Sodium in Patients With Severe Pulmonary Arterial Hypertension; NCT00147199).


Disruption of the blood-brain barrier by intra-arterial administration of papaverine: a technical note.
Various endovascular techniques can be used to treat cerebral vasospasm after aneurysmal subarachnoid haemorrhage (SAH) including intra-arterial administration of vasodilator drugs such as papaverine or nicardipine and balloon dilatation of the affected vessel segment. Papaverine is known to have side effects, and we report a possible new one. After the treatment of cerebral vasospasm in a SAH patient by intra-arterial administration of papaverine into the left posterior cerebral artery, severe mesencephalic extravasation of blood and contrast media was detected. After reviewing the literature, the authors conclude that interruption of the blood-brain barrier by papaverine most likely combined with a secondary hyperperfusion phenomena, and perhaps a direct toxic effect on brain tissue was the mechanism of this major complication. In treating vasospasm in areas with a high density of perforating arteries, especially in the posterior circulation, papaverine should be used cautiously because a safe regimen has yet to be established. In this situation, alternative agents such as calcium channel blockers could be considered, but evidence-based data are still missing.


Motion Planning Under Uncertainty In Highly Deformable Environments.
Many tasks in robot-assisted surgery, food handling, manufacturing, and other applications require planning and controlling the motions of manipulators or other devices that must interact with highly deformable objects. We present a unified approach for motion planning under uncertainty in deformable environments that maximizes probability of success by accounting for uncertainty in deformation models, noisy sensing, and unpredictable actuation. Unlike prior planners that assume deterministic deformations or treat deformations as a type of small perturbation, our method explicitly considers the uncertainty in large, time-dependent deformations. Our method requires a simulator of deformable objects but places no significant restrictions on the simulator used. We use a sampling-based motion planner in conjunction with the simulator to generate a set of candidate plans based on expected deformations. Our method then uses the simulator and optimal control to numerically estimate time-dependent state distributions based on uncertain parameters (e.g. deformable material properties or actuation errors). We then select the plan with the highest estimated probability of successfully avoiding obstacles and reaching the goal region. Using FEM-based simulation of deformable tissues, we demonstrate the ability of our method to generate high quality plans in two medical-inspired scenarios: (1) guiding bevel-tip steerable needles through slices of deformable tissue around obstacles for minimally invasive biopsies and drug-delivery, and (2) manipulating planar tissues to align interior points at desired coordinates for precision treatment.


Stability and movement of mRNAs and their encoded proteins in Xenopus oocytes.
The stability and movement of several polyadenylated (poly A+) and nonpolyadenylated (poly A-) mRNAs in Xenopus oocytes have been examined. At least 50% of the poly A+ mRNA molecules (9S rabbit globin mRNA, chicken ovalbumin, and lysozyme) were stable in oocytes over a 48-h period, irrespective of the amount injected. About 50% of injected poly A- reovirus mRNAs was degraded within the first 24 h of injection, irrespective of the amount injected, although no further degradation was observed over an additional 24 h. The movement of all poly A+ mRNAs injected at either the animal or vegetal pole of the oocyte was very slow. Little movement of RNA from the animal half to the vegetal half was observed even 48 h after injection. In contrast, similar amounts of mRNA were present in both halves 48 h after vegetal pole injection. Similar results were obtained after injection of poly A- reovirus mRNAs. The movement of the proteins encoded by the poly A+ mRNAs was studied in the 6-h period after injection when little mRNA movement had occurred. 85% of the globin synthesized accumulated in the animal half irrespective of injection site. The movement of the sequestered secretory proteins ovalbumin and lysozyme in the same oocytes as globin was much slower; very little lysozyme appeared in the half of the oocyte opposite the site of injection.


Serine-arginine protein kinase 1 (SRPK1) inhibition as a potential novel targeted therapeutic strategy in prostate cancer.
Angiogenesis is required for tumour growth and is induced principally by vascular endothelial growth factor A (VEGF-A). VEGF-A pre-mRNA is alternatively spliced at the terminal exon to produce two families of isoforms, pro- and anti-angiogenic, only the former of which is upregulated in prostate cancer (PCa). In renal epithelial cells and colon cancer cells, the choice of VEGF splice isoforms is controlled by the splicing factor SRSF1, phosphorylated by serine-arginine protein kinase 1 (SRPK1). Immunohistochemistry staining of human samples revealed a significant increase in SRPK1 expression both in prostate intra-epithelial neoplasia lesions as well as malignant adenocarcinoma compared with benign prostate tissue. We therefore tested the hypothesis that the selective upregulation of pro-angiogenic VEGF in PCa may be under the control of SRPK1 activity. A switch in the expression of VEGF165 towards the anti-angiogenic splice isoform, VEGF165b, was seen in PC-3 cells with SRPK1 knockdown (KD). PC-3 SRPK1-KD cells resulted in tumours that grew more slowly in xenografts, with decreased microvessel density. No effect was seen as a result of SRPK1-KD on growth, proliferation, migration and invasion capabilities of PC-3 cells in vitro. Small-molecule inhibitors of SRPK1 switched splicing towards the anti-angiogenic isoform VEGF165b in PC-3 cells and decreased tumour growth when administered intraperitoneally in an orthotopic mouse model of PCa. Our study suggests that modulation of SRPK1 and subsequent inhibition of tumour angiogenesis by regulation of VEGF splicing can alter prostate tumour growth and supports further studies for the use of SRPK1 inhibition as a potential anti-angiogenic therapy in PCa.


Temperature effects on the nanoindentation characterization of stiffness gradients in confined polymers.
The stiffening of polymers near inorganic fillers plays an important role in strengthening polymer nanocomposites, and recent advances in metrology have allowed us to sample such effects using local mechanical measurement techniques such as nanoindentation and atomic force microscopy. A general understanding of temperature and confinement effects on the measured stiffness gradient length-scale ξint is lacking however, which convolutes molecular interpretation of local property measurements. Using coarse-grained molecular dynamics and finite element nanoindentation simulations, we show that the measured ξint increases with temperature in highly confined polymer systems, a dependence which acts in the opposite direction in systems with low confinement. These disparate trends are closely related to the polymer's viscoelastic state and the resulting changes in incompressibility and dissipative ability as the polymer transitions from glassy to rubbery. At high temperatures above the glass transition temperature, a geometrically confined system restricts the viscous dissipation of the applied load by the increasingly incompressible polymer. The indentation causes a dramatic build-up of hydrostatic pressure near the confining surface, which contributes to an enlarged measurement of ξint. By contrast, a less-confined system allows the pressure to dissipate via intermolecular motion, thus lowering the measured ξint with increased temperature above the glass transition temperature. These findings suggest that the well-established thin film-nancomposite analogy for polymer mobility near interfaces can be convoluted when measuring local mechanical properties, as the viscoelastic state and geometric confinement of the polymer can affect the nanomechanical response during indentation purely from continuum effects.


Soluble type immune checkpoint regulators using multiplex luminex immunoassay in chronic hepatitis B patients.
Soluble immune checkpoint regulators (sICs) were reported to have clinical impact on the diagnosis and progress of various diseases. This study compared the serum levels of 16 sICs in patients with chronic hepatitis B (CHB) to elucidate their clinical significance. The sICs of 86 patients with CHB and 50 healthy controls (HCs) were measured using luminex-based multiplex assay. The sICs were correlated with laboratory markers and sIC levels were compared in cirrhotic and non-cirrhotic groups. The levels of soluble programmed death-ligand 1, soluble cluster of differentiation 80/B7-1 (sCD80/B7-1), soluble cluster of differentiation 86/B7-2, soluble B-lymphocyte and T-lymphocyte attenuator, soluble herpes virus entry mediator, soluble cluster 28, soluble cluster of differentiation 40, soluble glucocorticoid-induced TNFR-related protein, soluble ligand for receptor TNFRSF18/AITR/GITR, soluble Toll-like receptor 2 and soluble inducible T-cell costimulator (sICOS) were decreased, while soluble T-cell immunoglobulin and mucin-domain containing-3 (sTIM-3) was increased in patients with CHB. Soluble programmed cell death protein 1 and sTIM-3 both positively correlated with hepatitis B virus (HBV) DNA level and increased in entecavir or tenofovir used group. The sTIM-3 positively correlated with aspartate aminotransferase, alanine aminotransferase, gamma-glutamyl transferase and gamma-glutamyl transferase to platelet ratio and fibrosis-4. Soluble cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) decreased in the liver cirrhosis (LC) group compared with the non-LC group. sCD80/B7-1 decreased LC risk, while soluble lymphocyte-activation gene increased LC risk by logistic regression analysis. Our results showed the preliminary data on dysregulated sICs in patients with CHB that may have clinical significance in diagnosis of patients with CHB. It can be applied to develop therapeutic target of HBV infection.


Akt-mediated phosphorylation and activation of estrogen receptor alpha is required for endometrial neoplastic transformation in Pten+/- mice.
PTEN is a tumor suppressor gene frequently mutated in human cancers. In vitro and in vivo studies have shown that PTEN can exert its tumor suppressive function through a variety of mechanisms, including regulation of cell death and cell proliferation. However, it is still unclear which of the many downstream pathways are critical in each different tissue, in vivo. Loss of PTEN is the earliest detectable genetic lesion in the estrogen-related type I (endometrioid) endometrial cancer. Pten(+/-) mice develop endometrial neoplastic lesions with full penetrance, thus providing a model system to dissect the genetic and biochemical events leading to the transition from normal to hyperplastic and neoplastic endometrial epithelium. Here, we show that loss of Pten in the mouse endometrium activates Akt and results in increased phosphorylation of estrogen receptor alpha (ERalpha) on Ser(167). ERalpha phosphorylation results, in turn, in the activation of this nuclear receptor both in vivo and in vitro, even in the absence of ligand, and in its increased ability to activate the transcription of several of its target genes. Strikingly, reduction of endometrial ERalpha levels and activity dramatically reduces the neoplastic effect of Pten loss in the endometrium, in contrast to complete estrogen depletion. Thus, we provide for the first time in vivo evidence supporting the hypothesis that loss of Pten and subsequent Akt activation result in the activation of ERalpha-dependent pathways that play a pivotal role in the neoplastic process.


Community responses and perceived barriers to responding to child maltreatment.
Although child maltreatment has important effects on physical and psychological health, even serious cases often go unreported. Little is known about actions that individuals take when they know of an abused child, factors influencing whether they take action, or general population beliefs about how best to prevent maltreatment. A random-digit-dialed telephone survey of 504 Washington State civilian, English-speaking adults living in households with telephones was conducted in 2002. Respondents were asked whether they had ever known an abused child and if so, how they responded and any barriers they experienced to responding. Regardless of whether they had known an abused child, they were asked how they would respond in a hypothetical situation and hypothetical barriers. They were also asked what they believed to be effective in preventing maltreatment. Half (49% +/- 5%) of the respondents indicated that they had known a child they believed to be abused and of these, four-fifths (84% +/- 5%) indicated that they took some action, most frequently reporting the abuse to Child Protective Services, talking to the parents about the abuse or how to parent, or calling the police or other law enforcement. The most frequently reported barriers were fear of retaliation by the abusive parent, being afraid of making the child's situation worse, and not wanting to intrude on family privacy. About nine-tenths of respondents believed that mental health services and drug and alcohol treatment, support services such as food banks and crisis nurseries, and parenting education classes were effective in preventing abuse. These results provide evidence that most people are willing to intervene to help an abused child. However, barriers to intervening (particularly fear of retaliation) exist and may account for some of the failures to report abuse.


What to use to express the variability of data: Standard deviation or standard error of mean?
Statistics plays a vital role in biomedical research. It helps present data precisely and draws the meaningful conclusions. While presenting data, one should be aware of using adequate statistical measures. In biomedical journals, Standard Error of Mean (SEM) and Standard Deviation (SD) are used interchangeably to express the variability; though they measure different parameters. SEM quantifies uncertainty in estimate of the mean whereas SD indicates dispersion of the data from mean. As readers are generally interested in knowing the variability within sample, descriptive data should be precisely summarized with SD. Use of SEM should be limited to compute CI which measures the precision of population estimate. Journals can avoid such errors by requiring authors to adhere to their guidelines.


Utilizing nanoparticles for improving anti-biofilm effects of azithromycin: A head-to-head comparison of modified hyaluronic acid nanogels and coated poly (lactic-co-glycolic acid) nanoparticles.
The widespread resistance of bacteria to traditional antibiotic treatments has expedited the search for novel therapies against these pathogens. The hypothesis of this work is that two distinctively different polymeric delivery systems, specifically D-α-tocopherol polyethylene glycol 1000 succinate (TPGS)-poly(lactic-co-glycolic acid) (PLGA) nanoparticles and octenyl succinic anhydride-modified low molecular weight hyaluronic acid (OSA-HA) nanogels may be used to substantially improve the properties of azithromycin, allowing its use for effective treatment of Pseudomonas aeruginosa biofilm infections. Azithromycin was encapsulated in both delivery systems and the physicochemical properties of the loaded delivery systems, including size, surface charge and drug loading were evaluated. Additionally, particle interaction with a mucin layer, penetration into a bacterial biofilm, prevention of biofilm formation and eradication of pre-formed biofilms, the influence on production of virulence factors and bacterial motility as well as cytotoxicity towards hepatocytes and lung epithelial cells were compared head-to-head. The TPGS-PLGA nanoparticles noticeably improved the antimicrobial activity and the biofilm prevention activity of azithromycin whereas the OSA-HA nanogels showed reduced mucin interactions together with improved reduction of pre-formed biofilms and maintained the low eukaryotic cell cytotoxicity of azithromycin.


Unraveling Complex Local Protein Environments with 4-Cyano-l-phenylalanine.
We present a multifaceted approach to effectively probe complex local protein environments utilizing the vibrational reporter unnatural amino acid (UAA) 4-cyano-l-phenylalanine (pCNPhe) in the model system superfolder green fluorescent protein (sfGFP). This approach combines temperature-dependent infrared (IR) spectroscopy, X-ray crystallography, and molecular dynamics (MD) simulations to provide a molecular interpretation of the local environment of the nitrile group in the protein. Specifically, a two-step enantioselective synthesis was developed that provided an 87% overall yield of pCNPhe in high purity without the need for chromatography. It was then genetically incorporated individually at three unique sites (74, 133, and 149) in sfGFP to probe these local protein environments. The incorporation of the UAA site-specifically in sfGFP utilized an engineered, orthogonal tRNA synthetase in E. coli using the Amber codon suppression protocol, and the resulting UAA-containing sfGFP constructs were then explored with this approach. This methodology was effectively utilized to further probe the local environments of two surface sites (sites 133 and 149) that we previously explored with room temperature IR spectroscopy and X-ray crystallography and a new interior site (site 74) featuring a complex local environment around the nitrile group of pCNPhe. Site 133 was found to be solvent-exposed, while site 149 was partially buried. Site 74 was found to consist of three distinct local environments around the nitrile group including nonspecific van der Waals interactions, hydrogen-bonding to a structural water, and hydrogen-bonding to a histidine side chain.


Impact of molecular clustering inside nanopores on desorption processes.
Understanding the sorption kinetics of nanoporous systems is crucial for the development and design of novel porous materials for practical applications. Here, using a porous coordination polymer/quartz crystal microbalance (PCP/QCM) hybrid device, we investigate the desorption of various vapor molecules featuring different degrees of intermolecular (hydrogen bonding) or molecule-framework interactions. Our findings reveal that strong intermolecular interactions lead to the desorption process proceeding via an unprecedented metastable state, wherein the guest molecules are clustered within the pores, causing the desorption rate to be temporarily slowed. The results demonstrate the considerable impact of the chemical nature of an adsorbate on the kinetics of desorption, which is also expected to influence the efficiency of certain processes, such as desorption by gas purge.


Can staccato and interrupted/fractionated uroflow patterns alone correctly identify the underlying lower urinary tract condition?
Worldwide, uroflowmetry without simultaneous electromyography is often the only testing performed during the initial assessment of children with lower urinary tract symptoms. Various alterations in uroflow pattern are thought to indicate particular types of lower urinary tract conditions, specifically staccato uroflow indicating dysfunctional voiding and intermittent/fractionated uroflow indicating detrusor underactivity. We determined how reliable uroflow pattern alone is as a surrogate for simultaneously measured pelvic floor electromyography activity during voiding, and how well staccato and interrupted uroflow actually correlate with the diagnoses they are presumed to represent. We reviewed uroflow/electromyography studies performed during the initial evaluation of 388 consecutive neurologically and anatomically normal patients with persistent lower urinary tract symptoms. We identified those with staccato, interrupted/fractionated and mixed uroflow based on current International Children's Continence Society guidelines. A total of 69 girls (58.5%) and 49 boys (41.5%) met inclusion criteria. Staccato uroflow was noted in 60 patients, interrupted/fractionated uroflow in 28 and a combination in 30. An active electromyography during voiding confirmed the diagnosis of dysfunctional voiding in 33.3% of patients with staccato, 46.4% with interrupted/fractionated and 50% with mixed uroflow patterns. Diagnoses based on uroflow pattern appearance without simultaneous electromyography to support them can be misleading, and reliance on uroflow pattern alone can lead to overdiagnoses of dysfunctional voiding and detrusor underactivity. When assessing patients with uroflow, an accompanying simultaneous pelvic floor electromyography is of utmost importance for improving diagnostic accuracy and thereby allowing for the most appropriate therapy.


Placental diffusing capacity in unanesthetized rabbits.
The purpose of this study was to determine the placental diffusing capacity of CO in unanesthetized rabbits. A standard dose of CO was administered to 18 near-term rabbits, and at time intervals varying from 5 min to 4 h maternal and fetal blood samples were analyzed for carboxyhemoglobin saturation (HbCO). Following administration of CO, the maternal HbCO rose rapidly within the 1st min and remained nearly constant for the remainder of the experiment. The fetal HbCO after CO introduction to the mother rose as CO crossed the placenta from maternal to fetal blood. After 15 min the fetal HbCO rose to equal the maternal HbCO averaging 18.5%. The fetal HbCO after 1 h averaged 33% saturation, 1.7-fold greater than the maternal HbCO at 1 h. Thereafter there was little further fetal rise and apparently equilibrium was reached. The average placental diffusing capacity for CO was 2.3 ml/(min times mmHg times kg), a value fourfold higher than has previously been reported in anesthetized sheep and dogs. Possible reasons are species difference, effects of posture, influence of anesthesia and surgery, and effects of an open versus intact uterus.


Myoglobin in pelagic small cetaceans.
Although myoglobin (Mb) is considered to contribute significantly to the oxygen and diving capacity of marine mammals, few data are available for cetaceans. Cetacean by-catch in the tuna driftnet fisheries in the Sulu Sea, Philippines, afforded the opportunity to examine Mb content and distribution, and to determine muscle mass composition, in Fraser's (Lagenodelphis hosei) and spinner (Stenella longirostris) dolphins and a pygmy killer whale (Feresa attenuata). Age was estimated by body length determination. Stomach contents were analyzed for the presence or absence of milk and solid foods. It was hypothesized (a) that Mb concentration ([Mb]) would be higher in Fraser's and spinner dolphins than in other small cetaceans because of the known mesopelagic distribution of their prey, (b) that [Mb] would vary among different muscles according to function during diving, and (c) that [Mb] would increase with age during development. The results were as follows. (1) Myoglobin concentrations of the longissimus muscle in adult Fraser's (6.8-7.2 g 100 g-1 muscle) and spinner (5-6 g 100 g-1 muscle) dolphins and in an immature pygmy killer whale (5.7 g 100 g-1 muscle) were higher than those reported previously for small cetaceans. (2) [Mb] varied significantly among the different muscle types in adult dolphins but not in calves; in adults, swimming muscles had significantly higher [Mb] than did non-swimming muscles, contained 82-86 % of total Mb, and constituted 75-80 % of total muscle mass. (3) Myoglobin concentrations in Fraser's and spinner dolphins increased with size and age and were 3-4 times greater in adults than in calves. The high Mb concentrations measured in the primary locomotory muscles of these pelagic dolphins are consistent with the known mesopelagic foraging behaviour of Fraser's and spinner dolphins and suggest that the pygmy killer whale is also a deep-diving species. The high Mb concentrations in epaxial, hypaxial and abdominal muscle groups also support the primary locomotory functions suggested for these muscles in other anatomical studies. As in other species, the increase in [Mb] during development probably parallels the development of diving capacity.


Synchronous versus sequential chemo-radiotherapy in patients with early stage breast cancer (SECRAB): A randomised, phase III, trial.
The optimal sequence of adjuvant chemotherapy and radiotherapy for breast cancer is unknown. SECRAB assesses whether local control can be improved without increased toxicity. SECRAB was a prospective, open-label, multi-centre, phase III trial comparing synchronous to sequential chemo-radiotherapy, conducted in 48 UK centres. Patients with invasive, early stage breast cancer were eligible. Randomisation (performed using random permuted block assignment) was stratified by centre, axillary surgery, chemotherapy, and radiotherapy boost. Permitted chemotherapy regimens included CMF and anthracycline-CMF. Synchronous radiotherapy was administered between cycles two and three for CMF or five and six for anthracycline-CMF. Sequential radiotherapy was delivered on chemotherapy completion. Radiotherapy schedules included 40 Gy/15F over three weeks, and 50 Gy/25F over five weeks. The primary outcome was local recurrence at five and ten years, defined as time to local recurrence, and analysed by intention to treat. ClinicalTrials.gov NCT00003893. Between 02-July-1998 and 25-March-2004, 2297 patients were recruited (1150 synchronous and 1146 sequential). Baseline characteristics were balanced. With 10.2 years median follow-up, the ten-year local recurrence rates were 4.6% and 7.1% in the synchronous and sequential arms respectively (hazard ratio (HR) 0.62; 95% confidence interval (CI): 0.43-0.90; p = 0.012). In a planned sub-group analysis of anthracycline-CMF, the ten-year local recurrence rates difference were 3.5% versus 6.7% respectively (HR 0.48 95% CI: 0.26-0.88; p = 0.018). There was no significant difference in overall or disease-free survival. 24% of patients on the synchronous arm suffered moderate/severe acute skin reactions compared to 15% on the sequential arm (p < 0.0001). There were no significant differences in late adverse effects apart from telangiectasia (p = 0.03). Synchronous chemo-radiotherapy significantly improved local recurrence rates. This was delivered with an acceptable increase in acute toxicity. The greatest benefit of synchronous chemo-radiation was in patients treated with anthracycline-CMF. Cancer Research UK (CR UK/98/001) and Pharmacia.


Sex differences in glutamate receptor gene expression in major depression and suicide.
Accumulating data indicate that the glutamate system is disrupted in major depressive disorder (MDD), and recent clinical research suggests that ketamine, an antagonist of the N-methyl-d-aspartate (NMDA) glutamate receptor (GluR), has rapid antidepressant efficacy. Here we report findings from gene expression studies of a large cohort of postmortem subjects, including subjects with MDD and controls. Our data reveal higher expression levels of the majority of glutamatergic genes tested in the dorsolateral prefrontal cortex (DLPFC) in MDD (F21,59=2.32, P=0.006). Posthoc data indicate that these gene expression differences occurred mostly in the female subjects. Higher expression levels of GRIN1, GRIN2A-D, GRIA2-4, GRIK1-2, GRM1, GRM4, GRM5 and GRM7 were detected in the female patients with MDD. In contrast, GRM5 expression was lower in male MDD patients relative to male controls. When MDD suicides were compared with MDD non-suicides, GRIN2B, GRIK3 and GRM2 were expressed at higher levels in the suicides. Higher expression levels were detected for several additional genes, but these were not statistically significant after correction for multiple comparisons. In summary, our analyses indicate a generalized disruption of the regulation of the GluRs in the DLPFC of females with MDD, with more specific GluR alterations in the suicides and in the male groups. These data reveal further evidence that, in addition to the NMDA receptor, the AMPA, kainate and the metabotropic GluRs may be targets for the development of rapidly acting antidepressant drugs.


A cap 'n' collar protein isoform contains a selective Hox repressor function.
We have characterized a protein isoform (CncB) from the Drosophila cap 'n' collar locus that selectively represses cis-regulatory elements that are activated by the Hox protein Deformed. Of the three Cnc protein isoforms, CncB is expressed in a localized pattern in mandibular and labral cells of the head during mid-stages of embryogenesis. When CncB protein is absent or reduced, mandibular cells are homeotically transformed toward maxillary identities. This transformation is associated with persistent Deformed expression in anterior mandibular cells, since the Deformed autoactivation circuit is normally antagonized by CncB function in these cells. Heat-shock-induced ectopic expression of CncB in mid-stages of embryogenesis is sufficient to attenuate the activity of Dfd response elements in maxillary epidermal cells, but appears to have no effect in trunk epidermal cells on either the function or the response elements of other Hox proteins. CncB provides a mechanism to modulate the specificity of Hox morphogenetic outcomes, which results in an increase in the segmental diversity in the Drosophila head.


Genetic and antigenic characterization of Canadian invasive Neisseria meningitidis serogroup C (MenC) case isolates in the post-MenC conjugate vaccine era, 2009-2013.
We previously reported a shift in the electrophoretic type (ET) of invasive MenC in Canada from predominantly ET-15 to ET-37 in the post-MenC conjugate vaccine period. This study sought to confirm this trend by examining all culture-confirmed invasive MenC case isolates in Canada in the period from 1 January 2009 to 31 December 2013. Of the 50 MenC isolates, 18 belonged to ET-15, 28 belonged to ET-37 (but not ET-15), and four belonged to other clonal types. Analysis of the serotype and serosubtype antigens, porA and fetA gene sequences provided data to show that invasive MenC belonging to ET-15 and ET-37 were two very different subpopulations within the ST-11 clonal complex. Sequence analysis of the fHbp genes suggested that 12 different types of factor H-binding protein were found among the ET-15 isolates while 86 % of ET-37 isolates were found to have fHbp genes predicted to encode peptide 22. The nadA gene in 12 MenC isolates was disrupted due to IS1301 insertion and 11 of these 12 isolates belonged to ET-15. Ten per cent of the invasive MenC were found to have a frame-shift mutation in their fHbp genes that predicted no fHbp produced. Significant diversity and frame-shift mutations of fHbp genes were found in invasive MenC strains in Canada.


CD8+ T cell epitope-flanking mutations disrupt proteasomal processing of HIV-1 Nef.
CTL play a critical role in the control of HIV and SIV. However, intrinsic genetic instability enables these immunodeficiency viruses to evade detection by CTL through mutation of targeted antigenic sites. These mutations can impair binding of viral epitopes to the presenting MHC class I molecule or disrupt TCR-mediated recognition. In certain regions of the virus, functional constraints are likely to limit the capacity for variation within epitopes. Mutations elsewhere in the protein, however, might still enable immune escape through effects on Ag processing. In this study, we describe the coincident emergence of three mutations in a highly conserved region of Nef during primary HIV-1 infection. These mutations (R69K, A81G, and H87R) flank the HLA B*35-restricted VY8 epitope and persisted to fixation as the early CTL response to this Ag waned. The variant form of Nef showed a reduced capacity to activate VY8-specific CTL, although protein stability and expression levels were unchanged. This effect was associated with altered processing by the proteasome that caused partial destruction of the VY8 epitope. Our data demonstrate that a variant HIV genotype can significantly impair proteasomal epitope processing and substantiate the concept of immune evasion through diminished Ag generation. These observations also indicate that the scale of viral escape may be significantly underestimated if only intraepitope variation is evaluated.


Syndecan signaling: when, where and why?
Syndecans are the only family of transmembrane heparan sulphate proteoglycans. Invertebrates all appear to have one Syndecan core protein, but in mammals there are four. Examination of the core protein sequences shows that the cytoplasmic domains are the most conserved. This suggests that Syndecans make important interactions and/or signalling contributions. It has been established that all syndecans can interact with proteins of the actin-associated cytoskeleton, but details of signalling have been harder to ascertain. However, it appears that Syndecans can signal, primarily to the cytoskeleton, and the details are reviewed here. Only for vertebrate syndecan-4 is there substantial detail, and it remains a challenge to dissect the functions and signalling of other vertebrate and invertebrate syndecans.


Maps of in vivo oxygen pressure with submillimetre resolution and nanomolar sensitivity enabled by Cherenkov-excited luminescence scanned imaging.
Low signal-to-noise ratios and limited imaging depths restrict the ability of optical-imaging modalities to detect and accurately quantify molecular emissions from tissue. Here, by using a scanning external X-ray beam from a clinical linear accelerator to induce Cherenkov excitation of luminescence in tissue, we demonstrate in vivo mapping of the oxygenation of tumours at depths of several millimetres, with submillimetre resolution and nanomolar sensitivity. This was achieved by scanning thin sheets of the X-ray beam orthogonally to the emission-detection plane, and by detecting the signal via a time-gated CCD camera synchronized to the radiation pulse. We also show with experiments using phantoms and with simulations that the performance of Cherenkov-excited luminescence scanned imaging (CELSI) is limited by beam size, scan geometry, probe concentration, radiation dose and tissue depth. CELSI might provide the highest sensitivity and resolution in the optical imaging of molecular tracers in vivo.


Myocardial Regional Interstitial Fibrosis is Associated With Left Intra-Ventricular Dyssynchrony in Patients With Heart Failure: A Cardiovascular Magnetic Resonance Study.
Left ventricular (LV) dyssynchrony is associated with poor prognosis in patients with heart failure (HF). The mechanisms leading to LV dyssynchrony are not fully elucidated. This study evaluates whether myocardium regional variation in interstitial fibrosis is associated with LV dyssynchrony. Forty-two patients with systolic heart failure (SHF), 76 patients with heart failure with preserved ejection fraction (HFpEF) and 20 patients without HF received cardiovascular magnetic resonance imaging (MRI) study. LV was divided into 18 segments by short-axis view. In each segment, regional extracellular volume fraction (ECV) and the time taken to reach minimum regional volume (Tmv) were derived. Intra-LV dyssynchrony were represented by maximum difference (Dysyn_max) and standard deviation (Dysyn_sd) of all Tmv. The results showed that among the covariates, only age (1.87, 95% CI: 0.61-3.13, p = 0.004) and ECV (3.77, 95% CI: 2.72-4.81, p < 0.001) were positively associated with Tmv. The results remained robust in certain subgroups. In conclusion, we demonstrated that LV myocardium regional variation in interstitial fibrosis is closely related to LV intra-ventricular dyssynchrony irrespective of the LV global function. These data might help explain the pathophysiology of LV dyssynchrony and it's underlying mechanisms leading to poor prognosis.


Behavioral and sex ratio modification of Japanese medaka (Oryzias latipes) in response to environmentally relevant mixtures of three pesticides.
We exposed Japanese medaka (Oryzias latipes) to environmentally relevant concentrations of azinphos-methyl, chlorothalonil, endosulfan, and mixtures of all three to determine if combinations of these pesticides result in additive, less-than-additive, or more-than-additive effects. Medaka were exposed from fertilization until 7 days posthatching, and end points included survival, time to hatch, size at 7 days posthatching, activity level (as measured by distance swam) and foraging ability at 3 weeks posthatching, and adult size, liver size, and sex ratio at 5 months posthatching. Although exposure to individual pesticides or pesticide mixtures did not affect survival, hatching time, or foraging ability, fry exposed to azinphos-methyl were significantly smaller at 1 week of age, and those exposed to chlorothalonil and a combination of the chemicals showed reduced activity. Adult sex ratios were biased toward females in all groups exposed to pesticides, with those exposed to azinphos-methyl, chlorothalonil, and the pesticide mixture departing significantly from an even sex ratio. There was no evidence of additive or synergistic effects of pesticide mixtures.


Physiology and bioenergetics of [NiFe]-hydrogenase 2-catalyzed H2-consuming and H2-producing reactions in Escherichia coli.
Escherichia coli uptake hydrogenase 2 (Hyd-2) catalyzes the reversible oxidation of H2 to protons and electrons. Hyd-2 synthesis is strongly upregulated during growth on glycerol or on glycerol-fumarate. Membrane-associated Hyd-2 is an unusual heterotetrameric [NiFe]-hydrogenase that lacks a typical cytochrome b membrane anchor subunit, which transfers electrons to the quinone pool. Instead, Hyd-2 has an additional electron transfer subunit, termed HybA, with four predicted iron-sulfur clusters. Here, we examined the physiological role of the HybA subunit. During respiratory growth with glycerol and fumarate, Hyd-2 used menaquinone/demethylmenaquinone (MQ/DMQ) to couple hydrogen oxidation to fumarate reduction. HybA was essential for electron transfer from Hyd-2 to MQ/DMQ. H2 evolution catalyzed by Hyd-2 during fermentation of glycerol in the presence of Casamino Acids or in a fumarate reductase-negative strain growing with glycerol-fumarate was also shown to be dependent on both HybA and MQ/DMQ. The uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP) inhibited Hyd-2-dependent H2 evolution from glycerol, indicating the requirement for a proton gradient. In contrast, CCCP failed to inhibit H2-coupled fumarate reduction. Although a Hyd-2 enzyme lacking HybA could not catalyze Hyd-2-dependent H2 oxidation or H2 evolution in whole cells, reversible H2-dependent reduction of viologen dyes still occurred. Finally, hydrogen-dependent dye reduction by Hyd-2 was reversibly inhibited in extracts derived from cells grown in H2 evolution mode. Our findings suggest that Hyd-2 switches between H2-consuming and H2-producing modes in response to the redox status of the quinone pool. Hyd-2-dependent H2 evolution from glycerol requires reverse electron transport.


Non-consensual sharing of personal sexually explicit imagery among young people in Australia: results from an online survey.
Consensual sharing of personal sexually explicit imagery (SEI) is part of young people's sexual practise; however, harms arise if content is shared without consent. Australians aged 15-29 years were recruited for an online survey. Participants indicated if they had ever sent SEI of themselves to someone else, received SEI directly from the person in the imagery and whether they thought it was illegal to forward SEI without consent. Participants reported whether anyone shared their SEI without permission, if they told people, made official reports or if there were consequences for perpetrator(s). Logistic regression was used to identify associations between victimisation, gender, age group, sexual identity and knowledge of SEI-related law. In total, 1007 participants (65% female, mean age 23 years, 67% heterosexual) were recruited; 63% sent personal SEI to another person, 71% received SEI from the person pictured and 77% correctly identified it is illegal to forward SEI without consent. Thirteen percent (n = 126) indicated another person forwarded personal SEI without consent. In univariate analysis, victimisation was associated with identifying as non-heterosexual (odds ratio = 1.51, confidence interval = 1.03-2.22), but was independent from age group, gender and knowledge. In multivariate analysis, sexual identity, age group, gender and knowledge were not significantly associated with victimisation. Among participants who experienced non-consensual sharing of personal SEI, 63% told friends, 10% told family, 93% made no official report and 94% reported no consequences for perpetrator(s). Initiatives are needed to promote legal rights and enable young people to seek support.


Correcting optical-axis calculation in polarization-sensitive optical coherence tomography.
Polarization-sensitive optical coherence tomography (PSOCT) has found many applications in imaging birefringence tissue samples. Polarization-sensitive detection is often implemented by utilizing a circularly polarized incident light and detecting the two orthogonal horizontal- and vertical-polarized interference components. However, the obtained optical axes images were inappropriately represented as depth-dependent periodic maps in all reported studies. A detailed analysis confirmed that this misrepresentation was caused by the accumulation of optical retardation with depth. A simple method was proposed to numerically correct this optical-axis calculation. Experimental studies in tendon tissue demonstrated that this method can be applied to map the 2-D optical-axis distributions in enface PSOCT images.


Digital and optical reconstruction of images from suboptical diffraction patterns.
Digital and optical reconstruction techniques are applied to synthetic holograms that are recorded at suboptical frequencies. The first section of the paper considers digital reconstruction that entails the application of the inverse diffraction transform to the diffraction pattern of an object illuminated with a suboptical source. Different cross sections of the object are displayed on a CRT in a sequential fashion. Image enhancement techniques are also employed in the process of digital reconstruction. The second section outlines a method for partially alleviating the longitudinal distortion that is inherent in optical reconstruction from synthetic holograms because of the difference between the recording and reconstructing wavelengths. The paper considers both the diffuse and nondiffuse illumination schemes and discusses the relative advantages and disadvantages of digital and optical reconstruction schemes.


Possible optical functions of the central core in lenses of trilobite eyes: spherically corrected monofocality or bifocality.
The function of the central core in lenses of certain schizochroal-eyed trilobites is unknown. To understand the possible optical function(s) of this central core, we performed computational ray-tracing on the lens in the schizochroal compound eyes of a Silurian Dalmanites trilobite. We computed the intensity of light focused by the lens versus the distance from the lower lens surface along the optical axis as functions of the refractive indices n(lu) and n(cc) of the lower lens unit and the central core. We determined those values of n(lu) and n(cc) that ensure that the studied central-cored trilobite lens is monofocal, bifocal, or trifocal. The sharpness (as the measure of the correction for spherical aberration) of these focal points was quantitatively studied. We show here that one of the possible optical functions of the central core could be the correction for spherical aberration, independently of the number (1, 2, or 3) of focal points. Another possible optical function of the core could be to ensure bifocality of the lens. In this case the peripheral lens region could have a given focal length and the central lens region could possess a longer or shorter focal length, if the refractive index n(cc) of the core is smaller or larger than the refractive index n(lu) of the upper lens unit. Finally, trifocality of the lenses can be considered only as a theoretical option, but by no means an optically optimally functioning possibility.


C-reactive protein levels and outcomes after statin therapy.
Statins lower the levels of low-density lipoprotein (LDL) cholesterol and C-reactive protein (CRP). Whether this latter property affects clinical outcomes is unknown. We evaluated relationships between the LDL cholesterol and CRP levels achieved after treatment with 80 mg of atorvastatin or 40 mg of pravastatin per day and the risk of recurrent myocardial infarction or death from coronary causes among 3745 patients with acute coronary syndromes. Patients in whom statin therapy resulted in LDL cholesterol levels of less than 70 mg per deciliter (1.8 mmol per liter) had lower event rates than those with higher levels (2.7 vs. 4.0 events per 100 person-years, P=0.008). However, a virtually identical difference was observed between those who had CRP levels of less than 2 mg per liter after statin therapy and those who had higher levels (2.8 vs. 3.9 events per 100 person-years, P=0.006), an effect present at all levels of LDL cholesterol achieved. For patients with post-treatment LDL cholesterol levels of more than 70 mg per deciliter, the rates of recurrent events were 4.6 per 100 person-years among those with CRP levels of more than 2 mg per liter and 3.2 events per 100 person-years among those with CRP levels of less than 2 mg per liter; the respective rates among those with LDL cholesterol levels of less than 70 mg per deciliter were 3.1 and 2.4 events per 100 person-years (P<0.001). Although atorvastatin was more likely than pravastatin to result in low levels of LDL cholesterol and CRP, meeting these targets was more important in determining the outcomes than was the specific choice of therapy. Patients who had LDL cholesterol levels of less than 70 mg per deciliter and CRP levels of less than 1 mg per liter after statin therapy had the lowest rate of recurrent events (1.9 per 100 person-years). Patients who have low CRP levels after statin therapy have better clinical outcomes than those with higher CRP levels, regardless of the resultant level of LDL cholesterol. Strategies to lower cardiovascular risk with statins should include monitoring CRP as well as cholesterol.


Abnormal perikaryal immunoreactivity to the phosphorylated heavy neurofilament unit in intracerebral basal forebrain transplants.
Grafts of Embryonic Day 14-15 basal forebrain tissue (medial septal/diagonal band nuclei) were transplanted into an aspirative fimbria-fornix cavity or the hippocampus of young adult rats. After extended periods of survival (1 and 2 years) the grafts were examined with immunocytochemical probes to identify specific types of neurons and assess the (spatial) distribution of the phosphorylated heavy neurofilament protein. Subpopulations of the long-term transplanted neurons expressed immunoreactivity to choline acetyl-transferase (CAT) and the low-affinity nerve growth factor receptor (192-IgG). Axons from the grafted neurons, visualized with the monoclonal antibody RT97 to the Mr 200,000 phosphorylated neurofilament unit, were observed to extend over the surfaces of the brain and connect with the host hippocampus. In subgroups of neurons without apparent axonal connections to the hippocampus, a change from axonal to cell body RT97 immunoreactivity was evident. A population of these neurons with abnormal neurofilament immunostaining in the soma was simultaneously identified as cholinergic with the CAT antibody. These studies indicate that abnormal changes can develop in the cytoskeleton of neurons in long-term intracerebral septal transplants. Although the reasons for this type of neurofilament modification in the grafted neurons are unknown, inappropriate terminal connections may be an important factor in the expression of this cytoskeletal change.


Long-lasting auditory gating deficit accompanied by GABA(B) receptor dysfunction in the hippocampus after early-life limbic seizures in rats.
In a previous study, we reported a rat model of early-life limbic seizures which resulted in a loss of GABA(B) receptor inhibition in the hippocampus. Since gating of auditory evoked potentials in the hippocampus (auditory gating) requires GABA(B) receptors and spatial behaviors depend on the hippocampus, we hypothesize that rats with early-life limbic seizures manifest deficits of auditory gating and spatial behaviors. Seizure rats were given a single injection of GABA(B) receptor antagonist CGP56999A (1-1.2 mg/kg i.p.) on postnatal day (PND) 15, which induced multiple limbic seizures in 8h; control rats were given saline injection. When tested at 3-9 weeks after seizure/control treatment, seizure as compared to control rats showed no difference in finding a hidden platform in the water maze, but were deficient in learning and maintaining consecutive criterion performance in the 8-arm radial arm maze. Auditory gating, as measured by paired-click (conditioning followed by test click) average auditory evoked potentials in the hippocampus, revealed a significant difference between seizure rats and controls. Seizure as compared to control rats showed an increased ratio of the test to conditioning click response as adolescents (50 days old) or adults (70 days old). Heterosynaptic electric paired-pulse depression of hippocampal population excitatory postsynaptic potential in freely moving rats, a measure of hippocampal GABA(B)-receptor mediated inhibition, was decreased in seizure as compared to control rats. Seizure as compared to control rats showed increased locomotor activity in a novel open field for the first 10 min, and decreased activity at 15-60 min. However, auditory prepulse inhibition, a measure of sensorimotor gating, revealed no difference between seizure and control rats. In conclusion, early-life limbic seizures induced a long-lasting deficit in auditory gating, likely caused by GABA(B) receptor-mediated inhibition loss in the hippocampus. Auditory gating loss is a symptom of schizophrenia, and thus GABA(B) receptor inhibition loss in the hippocampus provides a mechanism linking early-life seizures to a psychiatric symptom.


Development of a method to produce freeze-dried cubes from 3 Pacific salmon species.
Freeze-dried boneless skinless cubes of pink (Oncorhynchus gorbuscha), sockeye (Oncorhynchus nerka), and chum (Oncorhynchus keta) salmon were prepared and physical properties evaluated. To minimize freeze-drying time, the kinetics of dehydration and processing yields were investigated. The physical characteristics of the final product including bulk density, shrinkage, hardness, color, and rehydration kinetics were determined. Results showed that freeze-dried salmon cubes from each of the 3 Pacific salmon species can be produced with a moisture content of less that 10% and a(w) less 0.4 and freeze-drying time of 9 h. Processing yields ranged from 26% to 28.4%, depending on fish species. Shrinkage was less than 12% and rehydration of freeze-dried cubes was rapid. The value-added products developed have the potential to be utilized as ingredients for ready-to-eat soups, as snack food, salad topping, and baby finger-food.


A case for evolutionary genomics and the comprehensive examination of sequence biodiversity.
Comparative analysis is one of the most powerful methods available for understanding the diverse and complex systems found in biology, but it is often limited by a lack of comprehensive taxonomic sampling. Despite the recent development of powerful genome technologies capable of producing sequence data in large quantities (witness the recently completed first draft of the human genome), there has been relatively little change in how evolutionary studies are conducted. The application of genomic methods to evolutionary biology is a challenge, in part because gene segments from different organisms are manipulated separately, requiring individual purification, cloning, and sequencing. We suggest that a feasible approach to collecting genome-scale data sets for evolutionary biology (i.e., evolutionary genomics) may consist of combination of DNA samples prior to cloning and sequencing, followed by computational reconstruction of the original sequences. This approach will allow the full benefit of automated protocols developed by genome projects to be realized; taxon sampling levels can easily increase to thousands for targeted genomes and genomic regions. Sequence diversity at this level will dramatically improve the quality and accuracy of phylogenetic inference, as well as the accuracy and resolution of comparative evolutionary studies. In particular, it will be possible to make accurate estimates of normal evolution in the context of constant structural and functional constraints (i.e., site-specific substitution probabilities), along with accurate estimates of changes in evolutionary patterns, including pairwise coevolution between sites, adaptive bursts, and changes in selective constraints. These estimates can then be used to understand and predict the effects of protein structure and function on sequence evolution and to predict unknown details of protein structure, function, and functional divergence. In order to demonstrate the practicality of these ideas and the potential benefit for functional genomic analysis, we describe a pilot project we are conducting to simultaneously sequence large numbers of vertebrate mitochondrial genomes.