The use of vapor phase extraction in metabolic profiling of phytohormones and other metabolites.
Through complex networks of signaling interactions, phytohormones regulate growth, development, reproduction and responses to biotic and abiotic stress. Comprehensive metabolomic approaches, seeking to quantify changes in vast numbers of plant metabolites, may ultimately clarify these complex signaling interactions and consequently explain pleiotropic effects on plant metabolism. Synergistic and antagonistic phytohormone signaling interactions, referred to as crosstalk, are often considered at the level of transduction without proper consideration of synthesis or accumulation of phytohormones because of the limitation and difficulty in quantifying numerous signals. Significant progress has recently been made in the expansion of metabolic profiling and analysis of multiple phytohormones [Birkemeyer et al. (J. Chromatogr. A, 2003, 993, 89); Chiwocha et al. (Plant J., 2003, 35, 405); Müller et al. (Planta, 2002, 216, 44); Schmelz et al. (Proc. Natl Acad. Sci. USA, 2003, 100, 10552)]. We recently presented a novel metabolic profiling approach to the analysis of acidic phytohormones and other metabolites based on a simplistic preparation scheme and analysis by chemical ionization-gas chromatography/mass spectrometry. We now provide a detailed description of this vapor phase extraction technique and use pathogen infection of Arabidopsis with Pseudomonas syringae DC3000 to illustrate metabolic changes in salicylic acid, cinnamic acid, jasmonic acid, indole-3-acetic acid, abscisic acid, unsaturated C(18) fatty acids, 12-oxo-phytodienoic acid, and phytotoxin coronatine. Directions for further method expansion are provided and include issues of recovery, derivatization, range of accessible analytes, optimization, reproducibility and future directions.


The combined scapular/parascapular flap for the treatment of extensive electrical burns of the upper extremity.
The combined scapular parascapular flap was used for coverage of six high voltage electrical burn defects of upper extremity. The flaps resurfaced the exposed tendon, nerve, bone or blood vessels on the volar and dorsal side of the distal forearm and the hand. In all cases successful soft tissue coverage and wound healing was achieved. After the stabilisation of the wounds, secondary reconstructive procedures, such as nerve grafting, tendon grafting, tendon transfers were performed. Nerve healing and tendon gliding was found to be successful under the flap.


A Multidisciplinary Approach for Treating Bilateral Congenitally Missing Lateral Incisors.
To effectively treat congenitally missing teeth (CMT), a multidisciplinary approach involving both clinical and laboratory components is often not only beneficial but necessary. Such an approach frequently may involve the use of orthodontics, removable partial dentures (RPDs), fixed partial dentures, indirect adhesive restorations, or implant-supported restorations (ISRs), or a combination of these. In the case presented, an adult female patient had CMT at site Nos. 7 and 10 and was ready to have her existing RPD that she had worn for many years replaced and her smile enhanced. The treatment called for ISRs placed in the sites of the lateral incisors with lithium-disilicate crowns cemented to custom zirconia abutments. Lithium-disilicate veneers would be placed on adjacent teeth. However, in order to carry out the plan, limited fixed orthodontics would first be needed to create sufficient space for the ISR.


Hyperinsulinemia does not change atherosclerosis development in apolipoprotein E null mice.
To determine the contribution of hyperinsulinemia to atherosclerosis development. Apolipoprotein E (Apoe) null mice that had knockout of a single allele of the insulin receptor (Insr) gene were compared with littermate Apoe null mice with intact insulin receptors. Plasma insulin levels in Insr haploinsufficient/Apoe null mice were 50% higher in the fasting state and up to 69% higher during a glucose tolerance test, but glucose tolerance was not different in the 2 groups. C-peptide levels, insulin sensitivity, and postreceptor insulin signaling in muscle, liver, fat, and aorta were not different between groups, whereas disappearance in plasma of an injected insulin analog was delayed in Insr haploinsufficient/Apoe null mice, indicating that impaired insulin clearance was the primary cause of hyperinsulinemia. No differences were observed in plasma lipids or blood pressure. Despite the hyperinsulinemia, atherosclerotic lesion size was not different between the 2 groups at time points up to 52 weeks of age when measured as en face lesion area in the aorta, cross-sectional plaque area in the aortic sinus, and cholesterol abundance in the brachiocephalic artery. Hyperinsulinemia, without substantial vascular or whole-body insulin resistance and without changes in plasma lipids or blood pressure, does not change susceptibility to atherosclerosis.


Simulation model for dental arch shapes.
To develop a simulation model for dental arch shapes. Analysis of measurements of dental casts to determine a general second degree equation for the dental arches. Department of Human Anatomy and School of Computing and Informatics, University of Nairobi. The measurement of dental casts, 30 (15M and 15F) each from three Kenyan ethnic groups (Maasai, Kalenjin, Kikuyu), aged 12 years. The arches change their shapes from a parabola to an ellipse, governed by the boundary conditions at the position of the canine tooth, based on the general second degree equation for the conic sections. The simulation model graphically confirms the change from parabolic to elliptic shapes of dental arches with boundary conditions at the canine. This could be used to show the changes in dental arches for other ethnic groups.


A pilot study on the effects of almond consumption on DNA damage and oxidative stress in smokers.
The effects of almond consumption on DNA damage and oxidative stress among cigarette smokers were studied. Thirty healthy adult male regular smokers were randomly divided into three groups, 10 subjects per group. Group A (control group) did not receive any almonds. Subjects in Groups B and C received 3 oz and 6 oz (84 g and 168 g) of almonds each day respectively for 4 wk. Two known biomarkers for DNA damage, urinary 8-hydroxy-2'-deoxyguanosine (8-OH-dG) and single strand DNA breaks of peripheral blood lymphocytes, were measured by enzyme-linked immunosorbent assay and comet assay, respectively. In addition, plasma malondialdehyde (MDA) level, superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activities were measured as biomarkers for oxidative stress. The results showed lower levels of urinary 8-OH-dG and single strand DNA breaks in the two almond-treated groups as compared with the control group. Furthermore, MDA levels in the almond-treated groups were lower than the controls. However, no significant effects of almonds on SOD and GSH-Px activities were found. In conclusion, results from this pilot study indicate that almond consumption has preventive effects on oxidative stress and DNA damage caused by smoking. A larger, randomized, placebo-controlled clinical trial on almonds will be initiated in the near future.


Using FBRM to investigate the sewage sludge flocculation efficiency of cationic polyelectrolytes.
Polyelectrolyte flocculation is a commonly used method for sewage sludge conditioning. The rate and extent of water removal from the flocculated sludge depends on the properties of the polyelectrolytes. This study investigates the flocculation performance of four different cationic polyelectrolytes using an in-situ laser probe which uses focused beam reflectance measurements (FBRM). It is used to characterise the floc number and size distribution of the flocculated sludge at various polyelectrolyte doses. Results show that the FBRM technique is very successful in tracking the change in particle population and chord lengths during the sludge flocculation process. The FBRM offers an alternative method for optimising the flocculation system in both selecting the flocculant and determining its optimum dose. Both the reduction in particle count for particles of less than 10 mum and the flocculation efficiency defined from the amount of the original sludge distribution remaining after flocculation correlate well with dewatering performance.


Could Short-Term Perioperative High-Dose Atorvastatin Offer Antiarrhythmic and Cardio-Protective Effects in Rheumatic Valve Replacement Surgery?
To evaluate the role of prophylactic high-dose atorvastatin for prevention of postoperative atrial fibrillation (POAF), inflammatory response attenuation, and myocardial protection after valve replacement cardiac surgery. Randomized controlled trial. Assiut University Hospitals. Sixty-four adult patients undergoing cardiac valve replacement surgery. The participants were equally divided into 2 groups. Group S received 80 mg of atorvastatin (oral tablets), 12 and 2 hours preoperatively, and on the 2nd, 3rd, 4th, and 5th postoperative days. Control group C received placebo at the same time periods. The incidence of POAF, postoperative white blood cell count, serum C-reactive protein, interleukin 6, and troponin I. Group S patients showed a lower incidence of POAF compared with the placebo group (p = 0.031). The white blood cell count showed significant reductions in group S compared with group C on the second, third, fourth, and fifth postoperative days. The C-reactive protein level showed significant reductions on the third, fourth, and fifth postoperative days in group S compared with group C (p = 0.001, 0.001, and 0.001, respectively). The serum level of interleukin 6 showed a significant reduction on the fifth postoperative day in group S compared with group C (p = 0.001). There was no significant difference between the 2 groups regarding the troponin I level and inotropic score. Prophylactic use of high dose atorvastatin can decrease the incidence of POAF and attenuate the inflammatory process in adult patients undergoing isolated rheumatic cardiac valve replacement surgery.


Chemical synthesis of dendrotoxin-I: revision of the reported structure.
Dendrotoxin I (DTX-I) is a 60-residue peptide from the venom of the black mamba snake Dendroaspis polylepis, which binds to neuronal K+ channels. The structure reported previously for DTX-I was synthesized for the first time by a solution procedure. The synthetic product was confirmed to have the correct primary and disulfide structure determined by peptide mapping, sequence analysis and mass measurements. Comparison of synthetic DTX-I with the natural one by high-performance liquid chromatography and capillary zone electrophoresis, as well as by sequence analysis, revealed that the Asn residue at position 12 in the synthetic peptide was Asp in the natural product. Synthesis of DTX-I with Asp at position 12 gave a peptide identical with the natural product in all aspects. NMR analysis of synthetic [Asn12]- and [Asp12]-DTX-I also supported our findings that the Asn residue at position 12 in the DTX-I molecule should be revised as Asp. [Asn12]- and [Asp12]-DTX-I had very similar binding affinities when tested against radiolabeled dendrotoxin binding to rat brain synaptosomal membranes.


Gold nanorod vaccine for respiratory syncytial virus.
Respiratory syncytial virus (RSV) is a major cause of pneumonia and wheezing in infants and the elderly, but to date there is no licensed vaccine. We developed a gold nanorod construct that displayed the major protective antigen of the virus, the fusion protein (F). Nanorods conjugated to RSV F were formulated as a candidate vaccine preparation by covalent attachment of viral protein using a layer-by-layer approach. In vitro studies using ELISA, electron microscopy and circular dichroism revealed that conformation-dependent epitopes were maintained during conjugation, and transmission electron microscopy studies showed that a dispersed population of particles could be achieved. Human dendritic cells treated with the vaccine induced immune responses in primary human T cells. These results suggest that this vaccine approach may be a potent method for immunizing against viruses such as RSV with surface glycoproteins that are targets for the human immune response.


Deciphering starch quality of rice kernels using metabolite profiling and pedigree network analysis.
The physiological properties of rice grains are immediately obvious to consumers. High-coverage metabolomic characterization of the rice diversity research set predicted a negative correlation between fatty acid and lipid levels and amylose/total starch ratio (amylose ratio), but the reason for this is unclear. To obtain new insight into the relationships among the visual phenotypes of rice kernels, starch granule structures, amylose ratios, and metabolite changes, we investigated the metabolite changes of five Japonica cultivars with various amylose ratios and two knockout mutants (e1, a Starch synthase IIIa (SSIIIa)-deficient mutant and the SSIIIa/starch branching enzyme (BE) double-knockout mutant 4019) by using mass spectrometry-based metabolomics techniques. Scanning electron microscopy clearly showed that the two mutants had unusual starch granule structures. The metabolomic compositions of two cultivars with high amylose ratios (Hoshiyutaka and Yumetoiro) exhibited similar patterns, while that of the double-knockout mutant, which has an extremely high amylose ratio, differed. Rice pedigree network analysis of the cultivars and the mutants provided insight into the association between metabolic-trait properties and their underlying genetic basis in rice breeding in Japan. Multidimensional scaling analysis revealed that the Hoshiyutaka and Yumetoiro cultivars were Indica-like, yet they are classified as Japonica subpopulations. Exploring metabolomic traits is a powerful way to follow rice genetic traces and breeding history.


A Prospective Cohort Study of Intimate Partner Violence and Unprotected Sex in HIV-Positive Female Sex Workers in Mombasa, Kenya.
We conducted a prospective cohort study to test the hypothesis that intimate partner violence (IPV) is associated with unprotected sex in HIV-positive female sex workers in Mombasa, Kenya. Women completed monthly visits and quarterly examinations. Any IPV in the past year was defined as ≥1 act of physical, sexual, or emotional violence by the current or most recent emotional partner ('index partner'). Unprotected sex with any partner was measured by self-report and prostate specific antigen (PSA) test. Recent IPV was associated with significantly higher risk of unprotected sex (adjusted relative risk [aRR] 1.91, 95 % CI 1.32, 2.78, p = 0.001) and PSA (aRR 1.54, 95 % CI 1.17, 2.04, p = 0.002) after adjusting for age, alcohol use, and sexual violence by someone besides the index partner. Addressing IPV in comprehensive HIV programs for HIV-positive women in this key population is important to improve wellbeing and reduce risk of sexual transmission of HIV.


Work related musculoskeletal injuries sustained by Australian osteopaths: qualitative analysis of effects on practitioner health, clinical practice, and patient care.
There is limited literature that explores the experiences of osteopaths injured while engaging in clinical practice. Evidence from other similar health professions has described the numerous effects of work-related musculoskeletal injuries (WRMI). Work-related musculoskeletal injury refers to trauma to joints, ligaments, muscles and tendons resulting from injury sustained while undertaking work duties. This research aimed to gain a contextualised understanding of the experiences of osteopaths who have sustained a work-related musculoskeletal injury while performing clinical practice. This research used a descriptive qualitative design. Participants were recruited as part of a larger cross-sectional study. Thirteen Australian osteopaths who had sustained a work-related musculoskeletal injury consented to participate in semi-structured interviews during May and June 2016. Thematic analysis was used to elicit important themes from the interview transcripts that had been recorded and transcribed verbatim. The qualitative accounts provided by the participants were coded for the impacts of their injuries on work, home life and leisure activities. The participants provided detailed, contextual information about their injuries, including the contributing factors and the experience of living with a WRMI. The findings indicate that injured osteopaths often continue working because of financial commitments and their dedication to patient care. The participants offered insights into the challenges they faced due to the injury and the management strategies they used to deal with the impact on their work and personal life. The injuries were mostly unreported, the burden being carried by the participants and their families. This is the first research that explores the experiences of osteopaths who have sustained a WRMI. We anticipate that this research will encourage a broad and constructive discussion within the profession of the issues associated with WRMIs, including risk minimisation and injury prevention. Further research is warranted to understand the relationship between osteopaths training in ergonomics and injury prevention. This would lead to the development of guidelines and educational curricula addressing safe work for osteopaths.


Deaths from stroke in US children, 1979 to 1998.
Studies have documented declining mortality from stroke in adults over the past two decades, with black adults at greater risk of death from stroke than whites in all years. As these findings have been attributed to control of stroke risk factors that are less important in children, trends and demographics of childhood stroke mortality are of interest, but have not been explored. The authors analyzed death certificate data for ischemic and hemorrhagic stroke (subarachnoid hemorrhage [SAH] and intracerebral hemorrhage [ICH]) in children under 20 years of age in the United States for the years 1979 through 1998, covering approximately 1.5 billion person-years. Childhood mortality from stroke declined by 58% overall, with reductions in all major subtypes (ischemic stroke decreased by 19%; SAH, by 79%; ICH, by 54%). Black ethnicity was a risk factor for mortality from all stroke types (relative risk 1.74 for ischemic stroke; 1.76 for SAH; 2.06 for ICH; p < 0.0001 for all types). Male sex was a risk factor for mortality from SAH (relative risk 1.30, p < 0.0001) and ICH (relative risk 1.21, p < 0.0001), but not from ischemic stroke (relative risk 1.02, p = 0.76). Mortality from stroke in US children has decreased dramatically over the last 20 years. Black children are at greater risk of death from all stroke types than are white children. As control of known stroke risk factors is unlikely to account for declining stroke mortality and ethnic differences in children, unrecognized stroke risk factors may be important.


Proteomic analysis of the balance between survival and cell death responses in cisplatin-mediated ototoxicity.
Cisplatin, a widely used anticancer drug, preferentially damages outer hair cells (OHCs) of the inner ear. In this study, an antibody microarray was used to identify early changes in protein expression in the rat cochlea induced by cisplatin. Only small changes in hearing thresholds (4-34 dB elevation) were detected two days after cisplatin treatment (12 mg/kg). OHC function, measured by otoacoustic emissions, was slightly depressed (10 dB), and little or no receptor cell loss was observed. However, cisplatin induced large changes in the expression of 19 proteins involved in apoptosis, cell survival, or progression through the cell cycle. Fifteen of the proteins are novel to the study of the inner ear. Immunoblotting confirmed increases in the levels of the pro-survival activating transcription factor 2 (ATF2), of pro-apoptotic serine-threonine protein kinase, receptor interacting protein, and a 70/75 kDa nitrotyrosine bearing doublet of unknown function. Anti-nitrotyrosine antibodies localized these oxidatively damaged proteins to the stereocilia of OHCs, the Golgi-centrosome region of Hensen's cells, nuclei of outer pillar cells, and tunnel crossing fibers innervating OHCs. The results of this proteomic analysis reflect the commencement of ototoxic and cell survival responses before the observation of a significant functional or anatomical loss.


Do Follicles of Obese Patients Yield Competent Oocytes/Embryos?
This study evaluated the competency of oocytes/embryos derived from follicles >15 mm in diameter from obese patients, compared with nonobese patients. A cohort study was conducted in a single tertiary medical center between July 2018 and May 2019. Before ultrasound-guided follicular aspiration, follicles were measured and those with maximal dimensional size >15 mm were tracked. Microscopic examination of the follicular aspirates was performed by an embryologist. Each follicle aspirated was evaluated for oocyte maturation, oocyte fertilization, and embryo quality. 457 follicles were measured: 380 (83.2%) in nonobese and 77 (16.8%) in obese patients. No in-between group differences were observed in the causes of infertility, patients' demographics, or ovarian stimulation characteristics. Oocytes were achieved during aspiration from 277 (72.8%) and 54 (70.0%) of the nonobese and obese groups, respectively (p = 0.67). No in-between group differences were observed in fertilization (2PN/oocyte), top quality embryo (TQE) per zygote (2PN), and TQE per follicle. Oocyte recovery rate from follicles >15 mm is unrelated to patients' BMI. Moreover, the oocytes recovered from obese patients are competent yielding comparable zygote and TQE per follicle/oocyte, compared with nonobese patients. Further investigation is required to strengthen this finding.


Defining recurrence of nonmelanoma skin cancer after Mohs micrographic surgery: Report of the American College of Mohs Surgery Registry and Outcomes Committee.
Standardized definitions and methods of surveillance for local recurrence of nonmelanoma skin cancer are critical in determining cure rates attributed to treatment modalities. We sought to offer a standard definition of local recurrence after surgical treatment of nonmelanoma skin cancer and to propose an acceptable surveillance period and tracking methods. A literature search was performed for background definitions of local recurrence and tracking methods. The American College of Mohs Surgery (ACMS) Registry and Outcomes Committee then conducted a modified Delphi process to arrive at consensus definitions. We define local recurrence as a tumor with comparable histology, with contiguity to the surgical scar after treatment, and that arises within the area of the previously treated tumor. This project reports the results of a modified Delphi method process involving members of the ACMS. The model described may not be useful for nonexcision type treatments such as topical chemotherapy, electrodessication and curettage, or radiation treatment. Previous definitions of recurrence and surveillance methods after surgical treatment of nonmelanoma skin cancer are variable and nonstandard. We describe consensus standards for defining and tracking recurrence that should allow for consistent scientific evaluation and development of performance data in skin cancer outcomes registries.


A study of posttraumatic disorders in children who experienced an industrial disaster in the Briey region.
To study posttraumatic disorders in children who were directly and indirectly involved in an industrial disaster; to assess the respective impact of traumatism exposure, parental disorders and sociodemographic variables on the posttraumatic disorders of the children. The children were assessed with self-administered questionnaires (STAIC, CDI, IES) and questionnaires filled in by parents (CPRS, CBCL). Parents were assessed with the GHQ-28. Forty-three exposed children were compared with 44 children who were exposed to the same risk (indirectly exposed group) and with a control group of 50 unexposed children. The exposed group obtained significantly higher anxiety and trauma-related scores than the control group and the threatened group, as well as higher scores of behavioural symptoms and of parental disorders. Indirectly exposed children did not have higher rates of symptoms than control children. The younger exposed children exhibited the highest psychopathological scores. Low sociodemographic status was associated with more disorders. There were no differences on questionnaire scores between girls and boys. Children's disorders correlated with disorders in both parents; but this only accounted for part of the variance, a finding which supports the hypothesis of a direct impact of the trauma on the child, irrespective of parental clinical status, SES of the family, children's age and gender. Children's and parent's disorders interact in a complex fashion which needs further study.


Pox-Protein Public Private Partnership program and upcoming HIV vaccine efficacy trials.
The purpose of review is to provide an overview of the Pox-Protein Public Private Partnership (P5) and highlight the progress of the P5 program, including an upcoming HIV vaccine efficacy trial in South Africa. The RV144 Thai vaccine efficacy trial was the first to demonstrate that an HIV-1 vaccine can prevent HIV acquisition. The P5 vaccine regimen uses an ALVAC prime and protein boost modeled after the RV144 vaccine and adapted for the subtype C virus predominant in the southern African region. This regimen was recently tested in the HIV Vaccine Trials Network 100 phase 1/2a study in South Africa. Based on prospectively defined immunogenicity thresholds, criteria were met to support the launch of an efficacy study in late 2016. The aim of this phase 2b/3 trial will be to improve upon the results of RV144, with increased and more durable vaccine efficacy, to accelerate the potential licensure of a preventive vaccine in southern Africa. The planned P5 efficacy trial, HIV Vaccine Trials Network 702, is designed to test and prospectively define correlates of protection, if efficacious. A vaccine with modest efficacy, vaccine efficacy at least 50%, could have substantial public health impact and significantly decrease the incidence of new infections in heavily burdened areas of the world.


An open-source system for in planta gene stacking by Bxb1 and Cre recombinases.
The rapid development of crops with multiple transgenic traits arouses the need for an efficient system for creating stacked cultivars. Most major crops rely on classical breeding to introgress the transgene from a laboratory variety to the numerous cultivars adapted to different growing regions. Even with vegetative propagated crops, genetic crosses are conducted during varietal improvement prior to vegetative cloning. The probability to assort the 'x' number of transgenic loci into a single genome may seem trivial, (¼) (x) for a diploid species, but given the 'y' number of other nontransgenic traits that breeders also need to assemble into the same genome, the (¼) (x+y) probability for a 'breeding stack' could quickly make the line conversion process unmanageable. Adding new transgenes onto existing transgenic varieties without creating a new segregating locus would require site-specific integration of new DNA at the existing transgenic locus. Here, we tested a recombinase-mediated gene-stacking scheme in tobacco. Sequential site-specific integration was mediated by the mycobacteriophage Bxb1 integrase-catalyzed recombination between attP and attB sites. Transgenic DNA no longer needed after integration was excised by Cre recombinase-mediated recombination of lox sites. Site-specific integration occurred in ~10% of the integration events, with half of those events usable as substrates for a next round of gene stacking. Among the site-specific integrants, however, a third experienced gene silencing. Overall, precise structure and reproducible expression of the sequentially added triple traits were obtained at an overall rate of ~3% of the transformed clones--a workable frequency for the development of commercial cultivars. Moreover, since neither the Bxb1-att nor the Cre-lox system is under patent, there is freedom to operate.


Clostridium difficile colonization and infection in patients with hepatic cirrhosis.
The aim of this study was to investigate the toxigenic Clostridium difficile colonization (CDC, colonization with toxigenic C. difficile but without symptoms) and C. difficile infection (CDI, active C. difficile infection resulting in disease symptoms) in hepatic cirrhosis patients, identify the risk factors of CDC, and determine the correlation between CDC and CDI. The strains of toxigenic C. difficile were isolated from patients with hepatic cirrhosis within 48 h after admission, followed by multilocus sequence typing (MLST). Patients were divided into toxigenic CDC group and noncolonized (NC) group according to the colonization. Logistic regression analysis was performed to analyse the risk factors for the CDC. Besides, the CDI incidence was compared between the two groups. Colonization of toxigenic C. difficile was identified in 104 cases (19.8 %). Eighteen sequence types (STs) were identified, among which ST-3, ST-54, ST-35 and ST-37 were the predominant types. Child-Pugh class C(relative risk, RR, 3.025; 95 % CI: 1.410-6.488), decrease of prothrombin time activity (PTA) (RR 2.180; 95 % CI: 1.368-3.476), decrease of platelet (RR 2.746; 95 % CI: 0.931-8.103) and concurrent hepatic encephalopathy (RR 1.740; 95 % CI: 1.012-2.990) were identified as the risk factors for the hepatic cirrhosis patients with CDC. The CDI incidence in the CDC group was also significantly higher than that of the NC group (26.0 % vs 1.7 %, P<0.001). An carriage rate of 19.8 % was reported in the hepatic cirrhosis patients with C. difficile colonization. Child's class C, decrease of PTA and platelet, and concurrent hepatic encephalopathy were the risk factors for the hepatic cirrhosis patients with C. difficile colonization. Hepatic cirrhosis patients with C. difficile colonization were more susceptible to CDI.


Hot-tip laser angioplasty: a review of the Texas Heart Institute experience.
At the Texas Heart Institute, from October 1986 to October 1988, laser thermal angioplasty was used in conjunction with balloon angioplasty for treatment of 201 atherosclerotic vascular lesions in 162 patients. All patients had significant symptoms or impending limb loss. Most of the occlusions (148) were located in the superficial femoral artery; the clinical success rate in those cases was 80%. Similar success rates were noted in 25 iliac occlusions (77%) and 16 popliteal occlusions (88%). A small number of tibial vessels were also reopened successfully. Analysis of the results showed a correlation between success rates and lesion length. The success rate dropped from 79% for occlusions that were 10 cm and shorter, to 56% for occlusions ranging from 11 to 20 cm, and to 55% for occlusions longer than 20 cm. The incidence of perforations in most laser series is reflective of operator experience. Perforations occurred in 19 of 90 vessels (21%) in the 1st year of our experience and in 3 of 111 vessels (2.7%) in the 2nd year. Other complications included groin hematomas in 2 patients and embolization to the trifurcation vessels in 4. One patient died 8 hours after the procedure of complications unrelated to laser angioplasty. The safety and efficacy of "hot-tip" laser recanalization in the peripheral vasculature have been proved in early clinical experience. Some preliminary investigations have shown that laser-probe applications may be feasible in the coronary vasculature as well. Future applications of laser systems will depend on their proven ability to reduce restenosis rates and to reliably recanalize totally occluded vessels.


Association between clinician factors and a patient's wish to hasten death: terminally ill cancer patients and their doctors.
This study investigated the clinical factors associated with a wish to hasten death among patients with advanced cancer receiving palliative care, with a focus on the role of clinician-related factors. Patients were grouped into high- and low-scoring groups on the basis of their wish to hasten death; doctor-patient pairs were formed. Questionnaire data collected from patients and their treating doctors were subjected to multivariate analysis. Significant predictors of a high wish to hasten death in terminally ill patients from among treating clinicians included the clinician's perception of the patient's lower optimism and greater emotional suffering, the patient indicating a wish to hasten death, the doctor willing to assist the patient in hastening death (if requested and legal), and the doctor reporting less training in psychotherapy. When these variables were combined with patient factors identified in a previous study, the model significantly predicted a wish to hasten death with the following variables-patient factors: a higher perceived burden on others, higher depressive symptom scores, and lower family cohesion; physician factors: the doctor willing to assist the patient in hastening death (if requested and legal), the doctor's perception of lower levels of optimism and greater emotional distress in the patient, and the doctor having less training in psychotherapy; and the setting of care: recent admission to a hospice. The findings support the multifactorial influences on the wish to hasten death and suggest that the role of the clinician is a vital context within which the wish to hasten death should be considered.


Medical Mobile App Classification Using the National Institute for Health and Care Excellence Evidence Standards Framework for Digital Health Technologies: Interrater Reliability Study.
Clinical governance of medical mobile apps is challenging, and there is currently no standard method for assessing the quality of such apps. In 2018, the National Institute for Health and Care Excellence (NICE) developed a framework for assessing the required level of evidence for digital health technologies (DHTs), as determined by their clinical function. The framework can potentially be used to assess mobile apps, which are a subset of DHTs. To be used reliably in this context, the framework must allow unambiguous classification of an app's clinical function. The objective of this study was to determine whether mobile health apps could be reliably classified using the NICE evidence standards framework for DHTs. We manually extracted app titles, screenshots, and content descriptions for all apps listed on the National Health Service (NHS) Apps Library website on July 12, 2019; none of the apps were downloaded. Using this information, 2 mobile health (mHealth) researchers independently classified each app to one of the 4 functional tiers (ie, 1, 2, 3a, and 3b) described in the NICE digital technologies evaluation framework. Coders also answered contextual questions from the framework to identify whether apps were deemed to be higher risk. Agreement between coders was assessed using Cohen κ statistic. In total, we assessed 76 apps from the NHS Apps Library. There was classification agreement for 42 apps. Of these, 0 apps were unanimously classified into Tier 1; 24, into Tier 2; 15, into Tier 3a; and 3, into Tier 3b. There was disagreement between coders in 34/76 cases (45%); interrater agreement was poor (Cohen κ=0.32, 95% CI 0.16-0.47). Further investigation of disagreements highlighted 5 main explanatory themes: apps that did not correspond to any tier, apps that corresponded to multiple tiers, ambiguous tier descriptions, ambiguous app descriptions, and coder error. The current iteration of the NICE evidence standards framework for DHTs did not allow mHealth researchers to consistently and unambiguously classify digital health mobile apps listed on the NHS app library according to their functional tier.


Paradoxical hypotension during dobutamine infusion for myocardial perfusion scintigraphy.
Dobutamine as a predominant beta-1 agonist increases heart rate and myocardial contractility and at sufficient high doses, it also increases systolic blood pressure. This study was undertaken to describe instances of paradoxical hypotension during dobutamine infusion for TI-201 myocardial perfusion SPECT study and the relationship between scintigraphic findings and hypotension occurred during dobutamine infusion. In 201 consecutive patients unable to perform adequate exercise, dobutamine TI-201 myocardial SPECT was performed. Dobutamine was infused starting from 10 micrograms/kg/min increasing to 40 micrograms/kg/min. Paradoxical hypotension was defined as a decrease in systolic blood pressure > or = 20 mmHg compared with baseline study. Paradoxical hypotension was observed in 40 patients (Group A) out of 201 (19.9%) while no significant change in systolic blood pressure was detected in the remaining 161 patients (Group B). Mean maximum fall in systolic blood pressure was 39 +/- 18 mmHg (range: 20-90). In 33 of 40 patients (83%) with paradoxical hypotension, scintigraphy was normal compared to 131 (81%) of the remaining 161 patients. In patients of Group A, angiography, echocardiography and tilt table tests were performed in 13, 11 and 6 patients respectively. Nine of 13 angiographic evaluations (69%), 10 of 11 echocardiographic evaluations (91%), all of the tilt table tests were normal. Additionally, all of the patients of Group A were clinically followed up at least 6 months after the myocardial perfusion scintigraphy. None of the patients had a cardiac event except one patient during the follow-up period. Paradoxical hypotension during dobutamine infusion for myocardial scintigraphy is not an uncommon finding and up to 19.9% patients may develop such hypotension. To maximize test safety, precautions should be taken during dobutamine myocardial stress test, since remarkable decrease in systolic blood pressure may occur. Unlike hypotension occurring with exercise test, hypotension response to dobutamine is not always a marker for coronary artery disease.


Clinical efficacy of endoscopic pancreatic drainage for pain relief with malignant pancreatic duct obstruction.
This study was conducted to investigate the efficacy of pancreatic drainage for pain relief in advanced pancreatic cancer. Seventy-one patients with pancreatic carcinoma were divided into two groups: dilated and non-dilated pancreatic ducts. All patients underwent endoscopic retrograde cholangiopancreatography (ERCP), endoscopic biliary stenting and pancreatic stenting. Visual Analog Scale (VAS) scores, pain remission rates and survival time were evaluated during follow-up. The post-ERCP VAS score of the dilated group was lower than that of the non-dilated group at 1 and 3 months post-ERCP. There was no difference at 6 months. The pain remission rate in the dilated duct group was significantly higher than that in non-dilated duct group in 1 and 3 months post-ERCP. The median survival times were 8.17 and 8.22 months respectively. Endoscopic pancreatic drainage can relieve pain of advanced pancreatic cancer accompanied by safe dilation of the pancreatic duct.


Validation of MRC Centre MRI calf muscle fat fraction protocol as an outcome measure in CMT1A.
To translate the quantitative MRC Centre MRI protocol in Charcot-Marie-Tooth disease type 1A (CMT1A) to a second site; validate its responsiveness in an independent cohort; and test the benefit of participant stratification to increase outcome measure responsiveness. Three healthy volunteers were scanned for intersite standardization. For the longitudinal patient study, 11 patients with CMT1A were recruited with 10 patients rescanned at a 12-month interval. Three-point Dixon MRI of leg muscles was performed to generate fat fraction (FF) maps, transferred to a central site for quality control and analysis. Clinical data collected included CMT Neuropathy Score. Test-retest reliability of FF within individual healthy calf muscles at the remote site was excellent: intraclass correlation coefficient 0.79, limits of agreement -0.67 to +0.85 %FF. In patients, mean calf muscle FF was 21.0% and correlated strongly with disease severity and age. Calf muscle FF significantly increased over 12 months (+1.8 ± 1.7 %FF, p = 0.009). Patients with baseline FF >10% showed a 12-month FF increase of 2.9% ± 1.3% (standardized response mean = 2.19). We have validated calf muscle FF as an outcome measure in an independent cohort of patients with CMT1A. Responsiveness is significantly improved by enrolling a stratified patient cohort with baseline calf FF >10%.


Effect of desipramine on norepinephrine metabolism in humans: interaction with aging.
To determine whether differences in neuronal reuptake contribute to age-related changes of sympathetic nervous system activity, we compared norepinephrine (NE) release and metabolism during [3H]NE infusion and decay in six young (age 19-26 yr) and seven older (age 61-73 yr) healthy nonobese subjects. Subjects were studied on a control day and on a separate day after desipramine (DMI; 125 mg orally), a neuronal reuptake blocker. Compartmental analysis of plasma NE specific activity was used to determine several NE kinetic parameters. Plasma NE levels and NE spillover rates were higher in the elderly. Although plasma NE was unaffected by DMI in both age groups, both the metabolic clearance rate of NE from plasma and the rate of NE spillover into plasma fell in young and older groups during DMI. Furthermore, DMI dramatically lowered the mass of NE in the extravascular compartment and the rate of NE entry into the extravascular compartment. Thus neuronal uptake blockade has major effects on NE release as well as NE metabolism in humans. However, age-related differences in NE kinetics cannot be explained by differences in neuronal uptake.


[An analysis of prior antibiotic treatment on the impact of meningococcal disease in children of the Valencian Community. The Study Group of Invasive Diseases].
The objective of this study was to describe the effect of antibiotics given prior to hospitalization of children with meningococcal disease and to assess their relationship with disease outcome and microbiological isolation. A prospective surveillance system in all hospitals of the Community of Valencia was implemented. All cases of children less than 15 years of age with clinically suspected invasive disease and: 1) N. meningitidis isolated in a normally sterile body fluid; 2) positive capsular antigens in blood or CSF and a positive Gram stain; and 3) clinical diagnosis of an invasive N. meningitidis disease. In a two-year period 157 cases were reported. In 143 cases, data about antibiotic prescription prior to hospitalization was known. Of these, 24.5% had received antibiotics and none had received parenteral penicillin or cephalosporins. Oral antibiotics decreased bacterial isolation (p < 0.001) and did not modify outcome (p = 0.66). Oral antibiotics, not recommended for N. meningitidis diseases, did not modify prognosis, but decreased bacterial isolation, and therefore worsened clinical handling of the cases and epidemiological studies.


Distinct behaviors of neural stem and progenitor cells underlie cortical neurogenesis.
Neocortical precursor cells undergo symmetric and asymmetric divisions while producing large numbers of diverse cortical cell types. In Drosophila, cleavage plane orientation dictates the inheritance of fate-determinants and the symmetry of newborn daughter cells during neuroblast cell divisions. One model for predicting daughter cell fate in the mammalian neocortex is also based on cleavage plane orientation. Precursor cell divisions with a cleavage plane orientation that is perpendicular with respect to the ventricular surface (vertical) are predicted to be symmetric, while divisions with a cleavage plane orientation that is parallel to the surface (horizontal) are predicted to be asymmetric neurogenic divisions. However, analysis of cleavage plane orientation at the ventricle suggests that the number of predicted neurogenic divisions might be insufficient to produce large amounts of cortical neurons. To understand factors that correlate with the symmetry of cell divisions, we examined rat neocortical precursor cells in situ through real-time imaging, marker analysis, and electrophysiological recordings. We find that cleavage plane orientation is more closely associated with precursor cell type than with daughter cell fate, as commonly thought. Radial glia cells in the VZ primarily divide with a vertical orientation throughout cortical development and undergo symmetric or asymmetric self-renewing divisions depending on the stage of development. In contrast, most intermediate progenitor cells divide in the subventricular zone with a horizontal orientation and produce symmetric daughter cells. We propose a model for predicting daughter cell fate that considers precursor cell type, stage of development, and the planar segregation of fate determinants.


Virological outcome after structured interruption of antiretroviral therapy for human immunodeficiency virus infection is associated with the functional profile of virus-specific CD8+ T cells.
A clear understanding of the antiviral effects of CD8(+) T cells in the context of chronic human immunodeficiency virus (HIV) infection is critical for the development of prophylactic vaccines and therapeutics designed to support T-cell-mediated immunity. However, defining the potential correlates of effective CD8(+) T-cell immunity has proven difficult; notably, comprehensive analyses have demonstrated that the size and shape of the CD8(+) T-cell response are not necessarily indicative of efficacy determined by measures of plasma viral load. Here, we conducted a detailed quantitative and qualitative analysis of CD8(+) T-cell responses to autologous virus in a cohort of six HIV-infected individuals with a history of structured interruption of antiretroviral therapy (ART) (SIT). The magnitude and breadth of the HIV-specific response did not, by themselves, explain the changes observed in plasma virus levels after the cessation of ART. Furthermore, mutational escape from targeted epitopes could not account for the differential virological outcomes in this cohort. However, the functionality of HIV-specific CD8(+) T-cell populations upon antigen encounter, determined by the simultaneous and independent measurement of five CD8(+) T-cell functions (degranulation and gamma interferon, macrophage inflammatory protein 1beta, tumor necrosis factor alpha, and interleukin-2 levels) reflected the emergent level of plasma virus, with multiple functions being elicited in those individuals with lower levels of viremia after SIT. These data show that the quality of the HIV-specific CD8(+) T-cell response, rather than the quantity, is associated with the dynamics of viral replication in the absence of ART and suggest that the effects of SIT can be assessed by measuring the functional profile of HIV-specific CD8(+) T cells.


Magnetic chitosan nanocomposite: Fabrication, properties, and optimization for adsorptive removal of crystal violet from aqueous solutions.
Recovery of crystal violet (CV) dye was investigated using magnetic chitosan nano-composites (MCNCs) and the effects of process variables (contact time, initial CV concentration, adsorbent dose, and pH) were optimized through response surface methodology. The reliability of the RSM models (first-order model, first model with interaction, the second-order model, and reduced model) was tested by fitting the data. A comparative analysis of the results derived from the models demonstrated that the reduced model was the best. According to modelling results, MCNCs dosage and contact time were found to be the most effective variables on the adsorption efficiency procedure, respectively. Also, pH had no significant effect on the adsorption uptake statistically. MCNC has the maximum adsorption efficiency (72%) when the contact time, adsorbent dosage, and initial concentration of CV were optimally set as 140 min, 1 g, and 77 mg/L, respectively. Quantity uptake of CV was evaluated using the Langmuir, Freundlich, and Temkin models. Based on findings, Freundlich isotherm fitted well with the experimental results. Kinetic studies showed that the pseudo-first-order model fitted the best the experimental data, which indicated that the adsorption rate of CV molecules onto MCNC was time-dependent. The studies on the well regenerability of MCNC in addition to its high potentiality of cationic dyes removal make it an attractive adsorbent in terms of dye-containing wastewaters treatment.


Renal sympathetic nerves modulate glomerular ANP receptors and filtration.
We examined characteristics of atrial natriuretic peptide (ANP) receptors in glomeruli isolated from subacutely (3-5 days) denervated (DNX) and contralateral nondenervated (non-DNX) kidneys of normal rats (NL) and rats subjected to water deprivation for 48 h (WD). Total ANP receptor density in DNX kidneys of WD rats, measured by competitive inhibition binding between 125I-labeled ANP and ANP, was twofold higher than non-DNX kidneys (726 +/- 96 vs. 384 +/- 32 fmol/mg protein, P less than 0.05). Equilibrium association constant (Ka) was not significantly different (2.33 +/- 0.43 vs. 3.34 +/- 0.78 x 10(9) M-1). In NL rats, there was no difference in ANP receptor density between DNX and non-DNX kidneys (244 +/- 20 and 264 +/- 16 fmol/mg protein). Production of guanosine 3',5'-cyclic monophosphate (cGMP), a putative second messenger of ANP, in response to ANP (10(-7) M) in glomeruli isolated from DNX was significantly larger than non-DNX kidneys of WD rats. To determine whether these changes in ANP receptors have functional consequences in vivo, glomerular capillary ultrafiltration coefficient (Kf) was assessed by micropuncture technique in WD Munich-Wistar rats. In DNX kidneys, ANP infusion (4 micrograms.kg-1.h-1) significantly increased whole kidney glomerular filtration rate (GFR) and single-nephron (SN) GFR (0.64 +/- 0.06 to 0.89 +/- 0.17 ml/min and 25 +/- 2 to 33 +/- 2 nl/min, respectively; n = 7) and Kf (1.26 +/- 0.29 to 2.18 +/- 0.41 nl.min-1.mmHg-1).(ABSTRACT TRUNCATED AT 250 WORDS)


Cardiac Baroreflex Variability and Resetting during Sustained Mild Effort.
This exploratory study assessed the pattern of closed-loop baroreflex resetting using multi-logistic-curve analysis. Operating point gain and ranges of RR-interval (RRI) and systolic blood pressure (SBP) are derived to examine how these relate to sympathetic activation. Sustained low-intensity isometric handgrip exercise, with a period of post-exercise circulatory occlusion (PECO), provided a model to study baroreflex resetting because the progression toward fatigue at constant tension induces a continuous increase in volitional contribution to neuro-cardiovascular control. Continuous measurements of muscle sympathetic nerve activity (MSNA), blood pressure, and RRI were made simultaneously throughout the experimental session. Spontaneous sequence analysis was used to detect episodes of baroreflex "engagements", but the results are examined with a view to the fundamental difference between experimental conditions that isolate the carotid sinus (open-loop) and intact physiological conditions (closed-loop). While baroreflex function under open-loop conditions can be described in terms of a single logistic curve, intact physiologic conditions require a family of logistic curves. The results suggest that the baroreflex is in a "floating" state whereby it is continuously resetting during the timeline of the experiment but with minute-by-minute average values that mimic the less complex step-wise resetting pattern reported under open-loop conditions. Furthermore, the results indicate that baroreflex function and resetting of the operating point gain is reflected not in terms of change in the values of blood pressure or RR-interval but in terms of change in the range of values of these variables prevailing under different experimental conditions.


The effects of prolonged epinephrine infusion on the physiology of the rainbow trout, Salmo gairdneri. II. Branchial solute fluxes.
Rainbow trout were infused continuously for 24 h with epinephrine in order to elevate circulating levels to those measured during periods of acute extracellular acidosis (about 5 X 10(-8) mol l-1). Concomitant effects on branchial solute fluxes were evaluated. Epinephrine infusion caused complex and differential adjustments of Na+ and Cl- unidirectional fluxes (influx and efflux) resulting in a significant elevation of the arithmetic difference between Na+ and Cl- net fluxes (JnetNa+-JnetCl-). A significant correlation existed between JnetNa+-JnetCl- and net branchial acid excretion (JnetH+), thereby suggesting a role for epinephrine in piscine acid-base regulation. The stimulation of JnetH+ by epinephrine was due primarily to a reduction in the excretion of titratable acid (JTA) accompanied by non-significant changes in ammonia excretion (JAmm). The results are discussed with respect to a role for epinephrine in regulating acid-base disturbances by interacting with branchial ionic exchange mechanisms.


Intraoperative and fluorescein angiographic findings of a secondary macular hole associated with age-related macular degeneration treated by pars plana vitrectomy.
Macular hole results from a tractional force at the vitreo-retinal interface which is developed by modification and subsequent degeneration of the posterior precortical vitreous and the internal limiting membrane (ILM). Meanwhile, the wet type of age-related macular degeneration (AMD) is caused by the submacular formation of choroidal neovascularization (CNV). Although exudative changes derived from CNV may cause epiretinal membrane (ERM) formation, which can also cause tractional force at the vitreo-retinal interface, there have been few reports of AMD-associated macular hole development in which the full thickness of the retinal tissue is completely torn by the tractional force. Moreover, intraoperative finding of macular hole associated with AMD with a possible involvement of subretinal lesion has not been described. A 78-year-old man diagnosed with wet AMD with subretinal fluid and mild cataract received 8 treatments with intravitreal pegaptanib. After AMD remission, he developed a secondary macular hole in the same eye. He underwent a pars plana vitrectomy that successfully closed the macular hole. Intraoperatively, it was found that the patient's vitreous was formed and that the ERM and ILM were adherent, suggesting the involvement of a tractional force at the vitreo-retinal interface due to an inflammatory reaction related to AMD and/or intravitreally injected chemical compounds, resulting in macular hole development. Changes in the condition of his AMD and the RPE were observed on a fluorescein angiogram (FA) and an indocyanine green angiogram (IA) that preceded macular hole development, suggesting that subretinal changes may also have been involved in the pathogenesis. These clinical data, including the intraoperative findings and the temporal changes in the angiograms, suggest that an inflammatory reaction at the vitreo-retinal interface and subretinal lesion related to AMD contribute to the macular hole development in AMD patients treated with intravitreal injection.


Song trait similarity in great tits varies with social structure.
For many animals, long-range signalling is essential to maintain contact with conspecifics. In territorial species, individuals often have to balance signalling towards unfamiliar potential competitors (to solely broadcast territory ownership) with signalling towards familiar immediate neighbours (to also maintain so-called "dear enemy" relations). Hence, to understand how signals evolve due to these multilevel relationships, it is important to understand how general signal traits vary in relation to the overall social environment. For many territorial songbirds dawn is a key signalling period, with several neighbouring individuals singing simultaneously without immediate conflict. In this study we tested whether sharing a territory boundary, rather than spatial proximity, is related to similarity in dawn song traits between territorial great tits (Parus major) in a wild personality-typed population. We collected a large dataset of automatized dawn song recordings from 72 unique male great tits, during the fertile period of their mate, and compared specific song traits between neighbours and non-neighbours. We show here that both song rate and start time of dawn song were repeatable song traits. Moreover, neighbours were significantly more dissimilar in song rate compared to non-neighbours, while there was no effect of proximity on song rate similarity. Additionally, similarity in start time of dawn song was unrelated to sharing a territory boundary, but birds were significantly more similar in start time of dawn song when they were breeding in close proximity of each other. We suggest that the dissimilarity in dawn song rate between neighbours is either the result of neighbouring great tits actively avoiding similar song rates to possibly prevent interference, or a passive consequence of territory settlement preferences relative to the types of neighbours. Neighbourhood structuring is therefore likely to be a relevant selection pressure shaping variation in territorial birdsong.


Persistent hypoglycemia is induced by tolbutamide administration in broiler chickens fed a low-carbohydrate diet.
With a view to gaining an insight into the regulatory mechanism of blood glucose concentrations specific to the chicken, an experimental induction of hypoglycemia was conducted by single or sequential administration of tolbutamide in broiler chickens fed a standard or low-carbohydrate diet. A single dosing of tolbutamide at levels of 25-200 mg/kg body weight decreased plasma glucose concentrations for 2 to 8 h after the dosing in chickens fed either diet. No significant rise in plasma insulin concentration was observed for 2 to 24 h after the single dosing of tolbutamide in chickens on either diet, with the exception of a significant rise when chickens on the standard diet received 100 mg tolbutamide. However, a transient increase of plasma insulin concentration was observed only in the 20 min immediately after the single dosing. Persistent hypoglycemia that was sustained for 5 days, with no significant changes in plasma insulin concentration, was induced by sequential dosing (3 times per day for 5 days, every 8 h) of tolbutamide (100 and 200 mg/kg body weight) in chickens fed the low-carbohydrate diet. In these chickens, the consistently low concentration of plasma glucose, with small diurnal variations, was evidenced by the determination of plasma glucose every 3 h in day 4/5 of the tolbutamide dosing. In chickens fed the standard diet, on the other hand, the low plasma glucose concentrations for 5 days were accompanied by significant diurnal fluctuations. Chickens with persistent hypoglycemia showed slight decreases in plasma non-esterified fatty acids (NEFA) concentration and only slight changes in blood D-3-hydroxybutyrate (3HB) concentration. The present study shows that the persistent hypoglycemia with normoinsulinemia, in the main, is induced by tolbutamide dosing in chickens fed a low-carbohydrate diet, and that the blood concentrations of NEFA and 3HB, alternatives of energy source in animals, are only slightly changed or not at all in hypoglycemic chickens.


Regulation of Mumps Virus Replication and Transcription by Kinase RPS6KB1.
Mumps virus (MuV) caused the most viral meningitis before mass immunization. Unfortunately, MuV has reemerged in the United States in the past several years. MuV is a member of the genus Rubulavirus, in the family Paramyxoviridae, and has a nonsegmented negative-strand RNA genome. The viral RNA-dependent RNA polymerase (vRdRp) of MuV consists of the large protein (L) and the phosphoprotein (P), while the nucleocapsid protein (NP) encapsulates the viral RNA genome. These proteins make up the replication and transcription machinery of MuV. The P protein is phosphorylated by host kinases, and its phosphorylation is important for its function. In this study, we performed a large-scale small interfering RNA (siRNA) screen targeting host kinases that regulated MuV replication. The human kinase ribosomal protein S6 kinase beta-1 (RPS6KB1) was shown to play a role in MuV replication and transcription. We have validated the role of RPS6KB1 in regulating MuV using siRNA knockdown, an inhibitor, and RPS6KB1 knockout cells. We found that MuV grows better in cells lacking RPS6KB1, indicating that it downregulates viral growth. Furthermore, we detected an interaction between the MuV P protein and RPS6KB1, suggesting that RPS6KB1 directly regulates MuV replication and transcription.IMPORTANCE Mumps virus is an important human pathogen. In recent years, MuV has reemerged in the United State, with outbreaks occurring in young adults who have been vaccinated. Our work provides insight into a previously unknown mumps virus-host interaction. RPS6KB1 negatively regulates MuV replication, likely through its interaction with the P protein. Understanding virus-host interactions can lead to novel antiviral drugs and enhanced vaccine production.


Risk of Acute Coronary Heart Disease After Sepsis Hospitalization in the REasons for Geographic and Racial Differences in Stroke (REGARDS) Cohort.
Sepsis is associated with long-term health consequences. We sought to determine the long-term risks of acute and fatal coronary heart disease (CHD) events after sepsis hospitalizations among community-dwelling adults. We analyzed data from 30329 participants in the population-based REasons for Geographic And Racial Differences in Stroke (REGARDS) cohort. Sepsis events included hospitalizations for a serious infection with ≥2 systemic inflammatory response syndrome criteria. Acute CHD events included myocardial infarctions (MIs; nonfatal and fatal) and acute CHD deaths. Fatal CHD included deaths ≤28 days of an acute MI and acute CHD deaths. We age- and time-matched each sepsis participant with 5 nonsepsis participants. We assessed the associations between sepsis hospitalizations and future acute and fatal CHD events using Cox regression, Gray's model, and competing risks analysis, adjusting for comorbidities. The matched cohort contained 1070 sepsis and 5350 nonsepsis participants. Risk of acute CHD was higher for sepsis than nonsepsis controls after adjusting for sex, race, education, income, region, tobacco use, and select chronic medical conditions (0-1 year adjusted hazard ratio [HR], 4.38 [95% confidence interval (CI), 2.03-9.45]; 1-4 years, 1.78 [1.09-2.88]; and 4+ years, 1.18 [0.52-2.67]). Risk of fatal CHD was similarly higher for sepsis than nonsepsis individuals (0-1 year adjusted HR, 3.12 [95% CI, 1.35-7.23]; 1-4 years, 3.29 [1.89-5.74]; and 4+ years HR, 1.15 [0.34-3.94]). The long-term risks of acute and fatal CHD are elevated after sepsis hospitalization. Management of acute CHD risk may be important for individuals surviving a sepsis event.


Pickering nano-emulsions stabilized by solid lipid nanoparticles as a temperature sensitive drug delivery system.
The development of biomaterials with low environmental impact has seen increased interest in recent years. In this field, lipid nanoparticles have found a privileged place in research and industry. The purpose of this study was to develop Pickering O/W nano-emulsions only stabilized by solid lipid nanoparticles (SLNs), as a new generation of safe, non-toxic, biocompatible, and temperature-sensitive lipid nano-carriers. The first part is dedicated to understanding the interfacial behavior of SLNs and their related stabilization mechanisms onto nano-emulsions formulated by ultrasonication. Investigations were focused on the surface coverage as a function of the SLN size and volume fraction of dispersed oil, in order to prove that the droplet stabilization is effectively performed by the nanoparticles, and to disclose the limitations of this formulation. Characterization is performed by dynamic light scattering and transmission electron microscopy. The second part of the study investigated SLN adsorption on a model oil/water interface (surface tension and rheology) through an axisymmetrical drop shape analysis (drop tensiometer), following the interfacial tension and the rheological behavior. The objective of this part is to characterize the phenomenon governing the droplet/interface interactions, and disclose the rheological behavior of the interfacial SLN monolayer. The effect of temperature was also investigated, proving a real destabilization of the nano-suspension when the sample is heated above a temperature threshold, impacting on the integrity of the SLNs, which partially melt, and strongly enhancing the release of a model drug (ketoprofen) encapsulated in the nano-emulsion oil core. To conclude, Pickering nano-emulsions only stabilized by SLNs appear to be a very efficient innovative drug nano-carrier, opening new doors as a potential temperature-sensitive drug delivery system.


Insights from the Northern Territory on factors that facilitate effective palliative care for Aboriginal peoples.
This article aims to assist remote communities to develop their own palliative care services by providing findings on successful strategies identified through a 2-year research project which developed an innovative model for Indigenous palliative care. The discussion is set in the context of an understanding of the notion of cultural safety and discusses the positive experiences of the benefits of palliative care from the perspectives of both consumers and health professionals. The findings show that successful outcomes are derived from generic factors associated with palliative care philosophy and practice and from more specific factors, including the provision of pragmatic support to overcome practical problems, and community visits by health professionals. Factors specific to cultural respect are important, including familiarity and continuity of health care providers, cultural respect for grieving practices, provision of comfort food and bush tucker, development of culturally appropriate built environments, use of traditional healers and respect for spiritual practices.


Ocular effects of oral lithium in humans.
Thirteen healthy adult subjects, 34.2 +/- 7.4 years of age received oral lithium carbonate, 900 or 1200 mg daily for 10 days. Refraction, visual acuity, visual fields, corneal sensation, basal tear secretion, ocular motility, convergence amplitude, near point of convergence, Hertel exophthalmometry, pupillary reflexes, intraocular pressure, voluntary and pilocarpine-induced accommodation, and biomicroscopy and ophthalmoscopy of the anterior and posterior ocular segments were determined 4 days prior to starting lithium, 10 days after commencing lithium, and 14 days after discontinuing lithium. Some subjective visual or ocular complaints were reported, but no clinically or statistically significant change occurred in any parameter studied. There were slight, non-significant tendencies toward constriction of the measured visual field, reduction of voluntary and pilocarpine-induced accommodation, and reduction of near convergence amplitude while taking lithium; the former two were reversed when lithium was discontinued, the latter was not.


Cauda equina syndrome.
Single or double-level compression of the lumbosacral nerve roots located in the dural sac results in a polyradicular symptomatology clinically diagnosed as cauda equina syndrome. The cauda equina nerve roots provide the sensory and motor innervation of most of the lower extremities, the pelvic floor and the sphincters. Therefore, in a fully developed cauda equina syndrome, multiple signs of sensory disorders may appear. These disorders include low-back pain, saddle anesthesia, bilateral sciatica, then motor weakness of the lower extremities or chronic paraplegia and, bladder dysfunction. Multiple etiologies can cause the cauda equina syndrome. Among them, non-neoplastic compressive etiologies such as herniated lumbosacral discs and spinal stenosis and spinal neoplasms play a significant role in the development of the cauda equina syndrome. Non-compressive etiologies of the cauda equina syndrome include ischemic insults, inflammatory conditions, spinal arachnoiditis and other infectious etiologies. The use of canine, porcine and rat models mimicking the cauda equina syndrome enabled discovery of the effects of the compression on nerve root neural and vascular anatomy, the impairment of impulse propagation and the changes of the neurotransmitters in the spinal cord after compression of cauda equina. The involvement of intrinsic spinal cord neurons in the compression-induced cauda equina syndrome includes anterograde, retrograde and transneuronal degeneration in the lumbosacral segments. Prominent changes of NADPH diaphorase exhibiting, Fos-like immunoreactive and heat shock protein HSP72 were detected in the lumbosacral segments in a short-and long-lasting compression of the cauda equina in the dog. Developments in the diagnosis and treatment of patients with back pain, sciatica and with a herniated lumbar disc are mentioned, including many treatment options available.


Carbamate as a potential anti-Alzheimer's pharmacophore: A review.
Alzheimer's disease (AD) is a progressive age-related neurodegenerative brain disorder, which leads to loss of memory and other cognitive dysfunction. The underlying mechanisms of AD pathogenesis are very complex and still not fully explored. Cholinergic neuronal loss, accumulation of amyloid plaque, metal ions dyshomeostasis, tau hyperphosphorylation, oxidative stress, neuroinflammation, and mitochondrial dysfunction are major hallmarks of AD. The current treatment options for AD are acetylcholinesterase inhibitors (donepezil, rivastigmine, and galantamine) and NMDA receptor antagonists (memantine). These FDA-approved drugs mainly provide symptomatic relief without addressing the pathological aspects of disease progression. So, there is an urgent need for novel drug development that not only addresses the basic mechanisms of the disease but also shows the neuroprotective property. Various research groups across the globe are working on the development of multifunctional agents for AD amelioration using different core scaffolds for their design, and carbamate is among them. Rivastigmine was the first carbamate drug investigated for AD management. The carbamate fragment, a core scaffold of rivastigmine, act as a potential inhibitor of acetylcholinesterase. In this review, we summarize the last 10 years of research conducted on the modification of carbamate with different substituents which primarily target ChE inhibition, reduce oxidative stress, and modulate Aβ aggregation.


A patient with Down syndrome with a de novo derivative chromosome 21.
Pure partial trisomy of chromosome 21 is a rare event. The patients with this aberration are very important for setting up precise karyotype-phenotype correlations particularly in Down syndrome phenotype. We present here a patient with Down syndrome with a de novo derivative chromosome 21. Karyotype of the patient was designated as 46,XY,der(21)(p13)dup(21)(q11.2q21.3)dup(21)(q22.2q22.3) with regard to cytogenetic, FISH and array-CGH analyses. Non-continuous monosomic, disomic and trisomic chromosomal segments through the derivative chromosome 21 were detected by array-CGH analysis. STR analyses revealed maternal origin of the de novo derivative chromosome 21. The dual-specificity tyrosine (Y)-phosphorylation regulated kinase 1A (DYRK1A) and Down Syndrome Critical Region 1 (DSCR1) genes that are located in Down syndrome critical region, are supposed to be responsible for most of the clinical findings of Down syndrome. However, our patient is the first patient with Down syndrome whose clinical findings were provided in detail, with a de novo derivative chromosome 21 resulting from multiple chromosome breaks excluding DYRK1A and DSCR1 gene regions.


Patient and public involvement in designing and conducting doctoral research: the whys and the hows.
Evidence shows that public and patient involvement in research has a positive effect on its quality and end-results. Thus, public and patient involvement in all stages of research is becoming commonplace. There are limited detailed examples however, that describe how to make this possible, especially for those doing PhD research. Doctoral researchers are often new to research practice or have limited experience and are often bound by strict time and financial constraints. It is also not usually a requirement of the award to involve public and patients in their research. Hence, they may not feel confident or motivated to involve or engage with public and patients during their research. We, four doctoral researchers, share examples from our own research studies that have included different approaches to public and patient involvement. Two studies formed public and patient advisory groups who helped design the research questions, data collection tools and recruitment methods. One enlisted the help of an online public and patient panel from a local hospital. A different study worked with patients from an established group to help define key medical words. We did face some challenges, such as the need to develop good group work skills and to apply for grants to cover reimbursement, but we all found it beneficial to involve patients in our studies. We noticed a positive effect on each study's progression and an improvement in our own self-esteem. In addition, having public and patient involvement helped reduce the isolation we felt as doctoral researchers. Thus, we strongly encourage more doctoral researchers to involve public and patients in their studies. Public and patient involvement (PPI) has been shown to have a positive impact on health and social care research. However, adequate examples describing how to operationalise effective PPI, especially in doctoral studies, are lacking. Hence, doctoral researchers new to research, or those with limited experience, can be discouraged from facilitating PPI in their research. This paper aims to describe and discuss in detail the approaches used by four doctoral researchers to incorporate PPI at different stages of their research studies from study design to disseminating findings.We aim to inform other doctoral researchers about the challenges and limitations relating to PPI that we faced. Through these, we share pragmatic recommendations for facilitating PPI during doctoral studies.The description of four case studies demonstrated that PPI could be incorporated at various stages during doctoral research. This has had a beneficial impact on our research study progression, researcher self-esteem and lastly, helped alleviate researcher isolation during doctoral studies.


Proteomic expression profiles of virulent and avirulent strains of Listeria monocytogenes isolated from macrophages.
Listeria monocytogenes is able to survive and proliferate within macrophages. In the current study, the ability of three L. monocytogenes strains (serovar 1/2a strain EGDe, serovar 4b strain F2365, and serovar 4a strain HCC23) to proliferate in the murine macrophage cell line J774.1 was analyzed. We found that the avirulent strain HCC23 was able to initiate an infection but could not establish prolonged infection within the macrophages. By contrast, strains EGDe and F2365 proliferated within macrophages for at least 7 h. We further analyzed these strains by comparing their protein expression profiles at 0 h, 3 h, and 5 h post-infection using multidimensional protein identification technology coupled with electrospray ionization tandem mass spectrometry. Our results indicated that similar metabolic and cell wall associated proteins were expressed by all three strains at 3 h post-infection. However, increased expression of stress response and DNA repair proteins was associated with the ability to proliferate in macrophages at 5 h post-infection. By comparing the protein expression patterns of these three L. monocytogenes strains during intracellular growth in macrophages, we were able to detect biological differences that may determine the ability of L. monocytogenes to survive in macrophages.


Hematopoietic stem cell transplantation in the leukodystrophies: a systematic review of the literature.
The objective of this study is to systematically review the literature on worldwide numbers of leukodystrophy patients undergoing hematopoietic stem cell transplantation (HSCT) as well as the safety and efficacy of the procedure in this patient population. A PubMed and EMBASE search up to June 2012 was conducted with a manual search of references from relevant articles. Selected studies were evaluated using internationally accepted criteria. The effect estimates of HSCT upon survival in early-stage disease versus late-stage disease were compared. One hundred and fifty-two studies qualified for inclusion and reported on a total of 689 patients. Study quality ranged from poor to good; no study was rated excellent. Small sample sizes limited most studies. Meta-analysis in a subset of larger studies indicates that transplantation in earlier stages of disease fairs better than in the late stages. Beyond survival, little longitudinal data on functional outcome is reported and neurological outcome is sparse. Further studies are needed to determine the neurological outcome following HSCT in the leukodystrophies. HSCT in the early stages of cerebral disease is still recommended for select leukodystrophies.


Use of acute-stage-specific antigens of Toxoplasma gondii for serodiagnosis of acute toxoplasmosis.
Antisera to acetone-fixed tachyzoites were prepared by immunizing rabbits intravenously with the fixed organisms. Immunoblots in which purified immunoglobulin G (IgG) of the antisera was used recognized four antigens of a supernatant (after centrifugation at 10,000 x g for 30 min) fraction of sonicated tachyzoites. A purified toxoplasma antigen fraction, referred to as acute-stage-specific (AC) antigen, that contained mainly three antigens was obtained by affinity chromatography by using the purified IgG. One antigen had a molecular weight of approximately 52,000, and two antigens had molecular weights of approximately 6,000. An enzyme-linked immunosorbent assay (AC-ELISA) in which this purified antigen fraction, obtained by affinity chromatography, was used for detection of IgG antibody detected early acute infection in sera of patients with toxoplasmic lymphadenopathy. Ninety-two percent of sera obtained within 2 months after onset of lymphadenopathy were positive in the AC-ELISA, whereas only 9% of sera obtained later than 5 months after onset of the clinical illness were positive. The AC-ELISA appears highly sensitive and specific for diagnosis of acute toxoplasmosis.


Encephalitis with Autoantibodies against the Glutamate Kainate Receptors GluK2.
The objective of this study was to report the identification of antibodies against the glutamate kainate receptor subunit 2 (GluK2-abs) in patients with autoimmune encephalitis, and describe the clinical-immunological features and antibody effects. Two sera from 8 patients with similar rat brain immunostaining were used to precipitate the antigen from neuronal cultures. A cell-based assay (CBA) with GluK2-expressing HEK293 cells was used to assess 596 patients with different neurological disorders, and 23 healthy controls. GluK2-ab effects were determined by confocal microscopy in cultured neurons and electrophysiology in GluK2-expressing HEK293 cells. Patients' antibodies precipitated GluK2. GluK2 antibody-specificity was confirmed by CBA, immunoprecipitation, GluK2-immunoabsorption, and GluK2 knockout brain immunohistochemistry. In 2 of 8 samples, antibodies reacted with additional GluK2 epitopes present in GluK1 or GluK3; in both, the reactivity was abrogated after GluK2 immuno-absorption. Six of 8 patients developed acute encephalitis and clinical or magnetic resonance imaging (MRI) features of predominant cerebellar involvement (4 presenting as cerebellitis, which in 2 patients caused obstructive hydrocephalus), and 2 patients had other syndromes (1 with cerebellar symptoms). One of the samples showed mild reactivity with non-kainate receptors (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors [AMPAR] and N-methyl-D-aspartate receptors [NMDAR]) leading to identify 6 additional cases with GluK2-abs among patients with anti-AMPAR (5/71) or anti-NMDAR encephalitis (1/73). GluK2-abs internalized GluK2 in HEK293 cells and neurons; these antibody-effects were reversible in neurons. A significant reduction of GluK2-mediated currents was observed in cells treated with patients' GluK2 serum following the time frame of antibody-mediated GluK2 internalization. GluK2-abs associate with an encephalitis with prominent clinicoradiological cerebellar involvement. The antibody effects are predominantly mediated by internalization of GluK2. ANN NEUROL 2021;90:107-123.


[National level response to pandemic (H1N1) 2009].
The World Health Organization (WHO) announced the emergence of a novel influenza on April 24, 2009, and they declared pandemic on June 11. In Korea, the proportion of influenza-like illness and the consumption of antiviral agents peaked in early November. The government established the Central Headquarters for Influenza Control and operated the emergency response system. In the quarantine stations, we checked the body temperature and collected quarantine questionnaires from all the arrivals from infected countries. We also isolated the confirmed cases in the national isolation hospitals. However, as the community outbreaks were reported, we changed strategy from containment to mitigation. We changed the antiviral agent prescription guideline so that doctors could prescribe antiviral agents to all patients with acute febrile respiratory illness, without a laboratory diagnosis. Also the 470 designated hospitals were activated to enhance the efficacy of treatment. We vaccinated about 12 million people and manage the adverse event following the immunization management system. In 2010, we will establish additional national isolation wards and support hospitals to establish fever clinics and isolation intensive care unit (ICU) beds. We will also make a computer program for managing the national isolation hospitals and designated hospitals. We will establish isolation rooms and expand the laboratory in quarantine stations and we will construct a bio-safety level 3 laboratory in each province. In addition, we plan to construct a bio-safety level 4 laboratory at a new Korea Centers for Disease Control and Prevention (KCDC) facilities in Ossong.


Developmental origin of peripheral ciliary band neurons in the sea urchin embryo.
In the sea urchin larva, most neurons lie within an ectodermal region called the ciliary band. Our understanding of the mechanisms of specification and patterning of these peripheral ciliary band neurons is incomplete. Here, we first examine the gene regulatory landscape from which this population of neural progenitors arise in the neuroectoderm. We show that ciliary band neural progenitors first appear in a bilaterally symmetric pattern on the lateral edges of chordin expression in the neuroectoderm. Later in development, these progenitors appear in a salt-and-pepper pattern in the ciliary band where they express soxC, and prox, which are markers of neural specification, and begin to express synaptotagminB, a marker of differentiated neurons. We show that the ciliary band expresses the acid sensing ion channel gene asicl, which suggests that ciliary band neurons control the larva's ability to discern touch sensitivity. Using a chemical inhibitor of MAPK signaling, we show that this signaling pathway is required for proper specification and patterning of ciliary band neurons. Using live imaging, we show that these neural progenitors undergo small distance migrations in the embryo. We then show that the normal swimming behavior of the larvae is compromised if the neurogenesis pathway is perturbed. The developmental sequence of ciliary band neurons is very similar to that of neural crest-derived sensory neurons in vertebrates and may provide insights into the evolution of sensory neurons in deuterostomes.


TREM2 interacts with TDP-43 and mediates microglial neuroprotection against TDP-43-related neurodegeneration.
Triggering receptor expressed on myeloid cell 2 (TREM2) is linked to risk of neurodegenerative disease. However, the function of TREM2 in neurodegeneration is still not fully understood. Here, we investigated the role of microglial TREM2 in TAR DNA-binding protein 43 (TDP-43)-related neurodegeneration using virus-mediated and transgenic mouse models. We found that TREM2 deficiency impaired phagocytic clearance of pathological TDP-43 by microglia and enhanced neuronal damage and motor impairments. Mass cytometry analysis revealed that human TDP-43 (hTDP-43) induced a TREM2-dependent subpopulation of microglia with high CD11c expression and phagocytic ability. Using mass spectrometry (MS) and surface plasmon resonance (SPR) analysis, we further demonstrated an interaction between TDP-43 and TREM2 in vitro and in vivo as well as in human tissues from individuals with amyotrophic lateral sclerosis (ALS). We computationally identified regions within hTDP-43 that interact with TREM2. Our data highlight that TDP-43 is a possible ligand for microglial TREM2 and that this interaction mediates neuroprotection of microglia in TDP-43-related neurodegeneration.


Subdiffraction-resolution fluorescence imaging of proteins in the mitochondrial inner membrane with photoswitchable fluorophores.
Understanding the structural organization of biomolecules in cells, sub-cellular compartments or membranes requires non-invasive methods of observation that provide high spatial resolution. Recent advancements in fluorescence microscopy paved the way for novel super-resolution observations with an optical resolution well below the diffraction barrier of light. Here, we demonstrate that commercially available standard fluorescent probes, i.e. Alexa 647 labeled antibodies, can be used as efficient photoswitches. In combination with localization microscopy approaches the method is ideally suited to study the spatial organization of proteins in sub-cellular structures and membranes. The simplicity of the method lies in the fact that standard immunocytochemistry assays together with photoswitchable carbocyanine fluorophores and conventional total internal reflection fluorescence (TIRF) microscopy can be used to achieve a lateral resolution of ~ 20nm. We demonstrate subdiffraction-resolution fluorescence imaging of intracellular F(0)F(1)-ATP synthase and cytochrome c oxidase in the inner membrane of mitochondria. Besides the high localization precision of individual proteins we demonstrate how quantitative data, i.e. the protein distribution in the membrane, can be derived and compared.


Nuclear fibroblast growth factor 2 (FGF2) isoforms inhibit bone marrow stromal cell mineralization through FGF23/FGFR/MAPK in vitro.
Fibroblast growth factor 23 (FGF23) is responsible for phosphate wasting and the phenotypic changes observed in human diseases such as X-linked hypophosphatemia (XLH). Targeted overexpression of nuclear high-molecular weight fibroblast growth factor 2 isoforms (HMW isoforms) in osteoblasts resulted in a transgenic mouse with phenotypic changes similar to XLH, including increased FGF23, hypophosphatemia, and rickets/osteomalacia. The goal of this study was to assess whether HMW isoforms also reduced mineralized bone formation via phosphate-independent effects in bone marrow stromal cells (BMSCs) by modulating FGF23/FGF receptor (FGFR)/extracellular signal-regulated kinase (ERK) signaling. To determine if decreased bone formation in BMSC cultures from HMW transgenic mice could be rescued by blocking this pathway, an FGF23 neutralizing antibody, the FGFR tyrosine kinase inhibitor SU5402 and the mitogen-activated protein kinase (MAPK) inhibitor PD98059 were used. FGF23 levels in the conditioned medium of HMW BMSC cultures were dramatically increased compared to BMSC from control (Vector) mice. Mineralized nodule formation was significantly decreased in HMW BMSC cultures compared with control cultures. The decreased nodule formation in HMW cultures was partially rescued by the FGF23 neutralizing antibody, SU5402 and PD98059. mRNA levels for the osteoblast-related genes, osteocalcin, Runt-related transcription factor 2 (Runx2), and osterix, and the osteocyte-related gene dentin matrix acidic phosphoprotein 1 (Dmp1) were significantly decreased in HMW cultures compared with control cultures, and the decreases were partially rescued by SU5402 or PD98059 treatment. Matrix-gla-protein (Mgp) mRNA was significantly higher in HMW cultures compared with control cultures, reduced by SU5402, but further increased by PD98059. Our results suggest that phosphate-independent effects of HMW isoforms in vitro may be directly mediated in part via FGF23 and that HMW isoforms signal via FGF23/FGFR/MAPK to inhibit bone formation in vitro.


Application of ovarian transposition during hysterectomy.
To study the optimal position and method for ovarian transposition and its benefits and indications. We performed ovarian transposition in 34 patients from August 1989 to December 2000. Twelve patients were diagnosed with stage Ib to IIa cervical cancer, 4 had stage Ia endometrial carcinoma, 12 had stage III to IV endometriosis, 4 had myoma of uterus, 1 had dysfunctional uterine bleeding, and 1 had an ovarian granulosa cell tumor. Surgery went as follows: the ovary was dissociated by clamp, the skin was incised and a tunnel was made, then the ovary was translocated to the subcutaneous site. In the cases of benign lesions, the ovarian vessel pedicel went in through the abdominal cavity, but in malignant tumors, it went out through the peritoneum. In both cases (benign lesions or malignant tumors), the short-term and long-term endocrine function of the translocated ovary remained normal. Furthermore, patients could supervise their translocated ovary themselves. Subcutaneous ovary transposition might prevent not only implantation of gastrointestinal cancer but also the extension of pelvic carcinoma to the ovary. Because of the shallow transposition and the incision scar, it is easy for patients to supervise themselves. Moreover, the site of the ovary is easy to locate for ultrasound examinations. Thus, it can obtain the goal of early prevention for cancer. Subcutaneous ovarian transposition with skin incision is the optimal selection and suitable for all patients with various gynecologic diseases in which ovary removal is not necessary.


The persistence of folly: critical examination of dissociative identity disorder. Part II. The defence and decline of multiple personality or dissociative identity disorder.
In this second part of our review, we continue to explore the illogical nature of the arguments offered to support the concept of dissociative identity disorder (DID). We also examine the harm done to patients by DID proponents' diagnostic and treatment methods. It is shown that these practices reify the alters and thereby iatrogenically encourage patients to behave as if they have multiple selves. We next examine the factors that make impossible a reliable diagnosis of DID--for example, the unsatisfactory, vague, and elastic definition of "alter personality." Because the diagnosis is unreliable, we believe that US and Canadian courts cannot responsibly accept testimony in favour of DID. Finally, we conclude with a guess about the condition's status over the next 10 years.


Miniaturized parallelized sandwich immunoassays.
This chapter describes the development and use of bead-based miniaturized multiplexed sandwich immunoassays for focused protein profiling. Bead-based protein arrays or suspension microarrays allow simultaneous analysis of a variety of parameters within a single experiment. In suspension microarrays capture antibodies are coupled onto color-coded microspheres. The applications of suspension microarrays are described, which allow to analyze proteins present in different types of body fluids, such as serum or plasma, cerebrospinal, pleural and synovial fluids, as well as cell culture supernatants. The chapter is divided into the generation of suspension microarrays, sample preparation, processing of suspension microarrays, validation of analytical performance, and finally pattern generation using bioinformatics tools.


[Inhibition of lactate and malate dehydrogenase by dyes containing vinylsulfone and chlorotriazine groups].
The difference spectra of lactate and malate dehydrogenase complexes with four native dyes containing vinylsulfonic and triazinic groups (light-resistant yellow 2KT, red-violet 2KT, etc.) were monitored in 0.1 M phosphate buffer pH 8.2 at 20 degrees C. The dissociation constants were calculated from the spectral data. The most stable complexes were lactate dehydrogenase--light-resistant yellow 2KT and malate dehydrogenase--light-resistant yellow 2KT ones. The values of delta H degree = 5.75 kcal/mole and standard thermodynamic parameters, delta G degree = -6.5 kcal/mole and delta S degree = 41.2 e. u., were calculated from the values of association constants for temperature dependence. The thermodynamic characteristics confirmed the key role of hydrophobic interactions in lactate dehydrogenase--reactive dye complex formation. All the dyes under study competitively inhibit lactate and malate oxidation by the corresponding dehydrogenases. The inhibition constants of both enzymes by the four dyes were determined at 20 degrees C in 0.1 M phosphate buffer pH 8.2. Light-resistant yellow 2KT appeared to be the most effective inhibitor of the enzymes.


The influence of NaYF₄:Yb,Er size/phase on the multimodality of co-encapsulated magnetic photon-upconverting polymeric nanoparticles.
We report the synthesis, characterisation and evaluation of the in vitro biocompatibility of polymeric nanoparticles with both magnetic and upconverting fluorescent properties. The particles consist of superparamagnetic iron oxide nanoparticles and upconverting NaYF4:Yb,Er nanoparticles co-encapsulated within a poly(glycidyl methacrylate) sphere. Two different upconverting nanoparticles (10 nm α-NaYF4:Yb,Er and 50 nm β-NaYF4:Yb,Er) were synthesised and the optical and magnetic properties of the composite polymeric nanoparticle systems assessed by near infra-red laser spectroscopy, SQUID magnetometry and proton relaxometry. A live-dead assay was used to assess the viability of PC-12 neural cells incubated with varying concentrations of the nanoparticles. The composite nanoparticles produced no observed impact on cellular viability even at concentrations as high as 1000 μg mL(-1). Confocal microscopy revealed uptake of nanoparticles by PC-12 cells and peri-nuclear cytoplasmic localisation. Both particle systems show favourable magnetic properties. However, only the nanospheres containing 50 nm β-NaYF4:Yb,Er were suitable for optical tracking because the presence of iron oxide within the composites imparts a significant quenching of the upconversion emission. This study demonstrates the size and phase of the upconverting nanoparticles are important parameters that have to be taken into account in the design of multimodal nanoparticles using co-encapsulation strategies.


Localization of papillofoveal bundles in primates.
Axons in the fovea are precisely organized to ensure accurate vision. We investigated the morphologic characteristics and localization of nerve bundles in the optic nerve in primates. Macaque eyes were studied for conventional and immunostaining, and also marmoset eyes for carbocyanine dye tracing. Locally confined lesions associated with similar findings to human age-related macular degeneration (ARMD) were also evaluated. Axons of retinal ganglion cells formed fasciculi near their origin, and these fasciculi formed bundles thereafter. In the retinal nerve fiber layer, ascending bundles assembled stratification adding proximal bundle underneath successively. Bundles in the arcuate zone displayed a characteristic fine, parallel arrangement, whereas those in the outside zone intermingled with undefined reticular bundles as they approached the optic nerve head. Macular bundles remained in groups and were distributed in the temporal wedge of the optic nerve head. Orthograde and retrograde tracing revealed that these bundles formed confined groups of various sizes and, ultimately, a specific group of small bundles located in the innermost row, near the central vessels. In addition, these bundles showed evidence of focal degenerative deterioration in eyes with ARMD. Papillomacular bundles have a characteristic alignment and configuration. Foveal bundles that compose the confined group closest to the optic trunk (which we term papillofoveal bundles) appear to have functional significance with respect to the isolated lesions that accompany central vision loss or preservation.


Role of CXCL9/CXCR3 chemokine biology during pathogenesis of acute lung allograft rejection.
Acute allograft rejection is a major complication postlung transplantation and is the main risk factor for the development of bronchiolitis obliterans syndrome. Acute rejection is characterized by intragraft infiltration of activated mononuclear cells. The ELR-negative CXC chemokines CXCL9, CXCL10, and CXCL11) are potent chemoattractants for mononuclear cells and act through their shared receptor, CXCR3. Elevated levels of these chemokines in bronchoalveolar lavage fluid have been associated with human acute lung allograft rejection. This led to the hypothesis that the expression of these chemokines during an allogeneic response promotes the recruitment of mononuclear cells, leading to acute lung allograft rejection. We performed studies in a rat orthotopic lung transplantation model of acute rejection, and demonstrated increased expression of CXCL9 and CXCL10 paralleling the recruitment of mononuclear cells and cells expressing CXCR3 to the allograft. However, CXCL9 levels were 15-fold greater than CXCL10 during maximal rejection. Inhibition of CXCL9 decreased intragraft recruitment of mononuclear cells and cellular expression of CXCR3, resulting in lower acute lung allograft rejection scores. Furthermore, the combination of low dose cyclosporin A with anti-CXCL9 therapy had more profound effects on intragraft leukocyte infiltration and in reducing acute allograft rejection scores. This supports the notion that CXCL9 interaction with cells expressing CXCR3 has an important role in the recruitment of mononuclear cells, a pivotal event in the pathogenesis of acute lung allograft rejection.