Predictors of health-related quality of life and costs in adults with epilepsy: a systematic review.
Given the high burden of epilepsy on both health-related quality of life (HRQoL) and costs, identification of factors that are predictive of either reduced HRQoL or increased expenditure is central to the better future targeting and optimization of existing and emerging interventions and management strategies for epilepsy. Searches of Medline, Embase, and Cochrane Library (up to July 2010) to identify studies examining the association between demographic, psychosocial, and condition-related factors and HRQoL, resource utilization or costs in adults with epilepsy. For each study, predictor factor associations were summarized on the basis of statistical significance and direction; the results were then combined across studies. Ninety-three HRQoL and 16 resource utilization/cost studies were included. Increases in seizure frequency, seizure severity, level of depression, and level of anxiety and presence of comorbidity were strongly associated with reduced HRQoL. The majority of studies were cross-sectional in design and had an overall methodologic quality that was judged to be "moderate" for HRQoL studies and "poor" for health care resource or costs studies. In the 53 multivariate studies, age, gender, marital status, type of seizure, age at diagnosis, and duration of epilepsy did not appear to be associated with HRQoL, whereas the predictive influence of educational and employment status, number of antiepileptic drugs (AEDs) and AED side effects was unclear. The association between predictive factors and HRQoL appeared to be consistent across individuals whether refractory or seizures controlled or managed by AEDs. There were insufficient multivariate studies (five) to reliably comment on the predictors of resource utilization or cost in epilepsy. In addition to seizure control, effective epilepsy management requires the early detection of those most at risk of psychological dysfunction and comorbidity, and the targeting of appropriate interventions. There is need for more rigorous studies with appropriate multivariate statistical methods that prospectively investigate the predictors of HRQoL, resource utilization, and costs in epilepsy.


Characterization of heat-hyperalgesia in an experimental trigeminal neuropathy in rats.
Secondary trigeminal neuralgia (STN) follows an injury to the trigeminal nerve or one of its branches. Although rare, this condition results in great suffering and it is notoriously difficult to treat. The experimental analysis of painful neuropathy due to damage to the innervation of the limbs (e.g., the sciatic nerve) has progressed rapidly in recent years, but very few reports have appeared concerning experimental neuropathy in the trigemenial region. We report here an experimental rat model of trigeminal neuropathic pain produced by a chronic constriction injury to the infraorbital nerve (CCI-ION), and on a method that detects heat-evoked pain-related behavior. Rats with the CCI-ION have clear signs of heat-hyperalgesia when stimulated on the snout (the vibrissal pad). The hyperalgesia is seen both ipsi- and contralateral to the side of nerve injury, but is significantly more severe ipsilaterally, and lasts about 12 days.


Early Prediction of Response to Tyrosine Kinase Inhibitors by Quantification of EGFR Mutations in Plasma of NSCLC Patients.
The potential to accurately quantify epidermal growth factor receptor (EGFR) mutations in plasma from non-small-cell lung cancer patients would enable more rapid and more frequent analyses to assess disease status; however, the utility of such analyses for clinical purposes has only recently started to explore. Plasma samples were obtained from 69 patients with EGFR-mutated tumors and 21 negative control cases. EGFR mutations in plasma were analyzed by a standardized allele-specific polymerase chain reaction (PCR) test and ultra-deep next-generation sequencing (NGS). A semiquantitative index (SQI) was derived from dilutions of known EGFR mutation copy numbers. Clinical responses were evaluated by Response Evaluation Criteria in Solid Tumors 1.1 criteria and expressed as percent tumor shrinkage. The sensitivity and specificity of the PCR test and NGS assay in plasma versus tissue were 72% versus 100% and 74% versus 100%, respectively. Quantitative indices by the PCR test and NGS were significantly correlated (p < 0.001). EGFR testing at baseline and serially at 4 to 60 days during tyrosine kinase inhibitor therapy revealed a progressive decrease in SQI, starting from day 4, in 95% of cases. The rate of SQI decrease correlated with percent tumor shrinkage at 2 months (p < 0.0001); at 14 days, it was more than 50% in 70% of patients (rapid responders). In two patients with slow response, an early increase in the circulating levels of the T790M mutation was observed. No early T790M mutations were seen in plasma samples of rapid responders. Quantification of EGFR mutations from plasma with a standardized PCR test is feasible. To our knowledge, this is the first study showing a strong correlation between the EGFR SQI in the first days of treatment and clinical response with relevant implications for patient management.


Control of intestinal promoter activity of the cellular migratory regulator gene ELMO3 by CDX2 and SP1.
An important aspect of the cellular differentiation in the intestine is the migration of epithelial cells from the crypt to the villus tip. As homeodomaine transcription factor CDX2 has been suggested to influence cell migration, we performed a genome-wide promoter analysis for CDX2 binding in the differentiated human intestinal cancer cell line Caco-2 in order to identify CDX2-regulated genes involved in cellular migration. The engulfment and cell motility 3 (ELMO3) gene was identified as a potential CDX2 target gene. ELMO3 is an essential upstream regulator of the GTP-binding protein RAC during cell migration. However, no information is available about the transcriptional regulation of the ELMO3 gene. The aim of this study was to investigate the potential role of CDX2 in the regulation of the ELMO3 promoter activity. Electrophoretic mobility shift assays showed that CDX2 bound to conserved CDX2 sequences and mutations of the CDX2-binding sites, significantly reduced the promoter activity. Reporter gene assays demonstrated that the region mediating ELMO3 basal transcriptional activity to be located between -270 and -31 bp. Sequence analysis revealed no typical TATA-box, but four GC-rich sequences. In vitro analyses (electrophoretic mobility shift assays and promoter analyses) demonstrate that the SP1-binding sites are likely to play an important role in regulating the ELMO3 promoter activity. Furthermore, we showed here that CDX2 and SP1 can activate the ELMO3 promoter. Taken together, the present study reports the first characterization of the ELMO3 promoter and suggests a significant role of CDX2 in the basal transcriptional regulation of the intestine-specific expression of ELMO3, possibly through interaction with SP1.


Diagnostic yield of MRI for audiovestibular dysfunction using contemporary referral criteria: correlation with presenting symptoms and impact on clinical management.
To investigate the diagnostic yield of T2-weighted magnetic resonance imaging (MRI) screening for vestibular schwannoma and other relevant conditions in the setting of audiovestibular symptoms, given the more liberal contemporary referral criteria. To determine whether presenting clinical symptoms correlate with imaging outcome in order to guide future protocols for MRI referral. Eight hundred and eighty-one consecutive MRI examinations performed in patients with audiovestibular dysfunction were reviewed. Clinical indications and findings were recorded. Case notes were reviewed in patients with positive imaging findings. Two-way, cross-tabulation, Chi-square analysis was performed to assess the relationship between presenting symptoms and imaging outcome. Twelve of the 881 (1.4%) were positive for vestibular schwannoma. A further four of 881 (0.4%) revealed other relevant conditions. Incidental conditions, felt to be irrelevant to the presenting symptoms, were noted in 12 of the 881 (1.4%). In all 12 cases that were positive for vestibular schwannoma, either tinnitus or hearing loss was present. The yield for T2-weighted MRI to diagnose vestibular schwannoma and other relevant retrocochlear conditions was lower than for previous studies, which is likely to reflect trends in referral criteria. No single audiovestibular symptom or combination of symptoms is a statistically significant predictor of imaging outcome.


[The cerebral control of afferent somatosensory projections].
Cortical and dorsal column nuclei somatosensory evoked potentials (SEP) induced by electrical stimulation of the median nerve were recorded and analysed in 16 healthy volunteers practising transcendental meditation (TM) for two years. The records were performed before and during TM. The SEP changes during TM consisted of an increase in early SEP components amplitude. There were no changes in early SEP components peak latencies during TM.


Instep island flaps.
The instep island flap is safe and useful. It is based on branches of the posterior tibial artery and can be a musculocutaneous or a direct fasciocutaneous island flap. Sensory branches of the digital nerves may be transposed with the flap. There is a wide arc of transposition and the flap has survived even in very difficult circumstances.


Bone marrow spontaneous lesions in rodents from nonclinical 104-week carcinogenicity studies.
The authors performed a retrospective study to determine the incidences and range of spontaneous lesions in the bone marrow (sternum and femur) of control mice and rats. Data was collected from 2186 mice (Crl:CD-1(ICR)BR), and 2347 rats (Han Wistar and CD(SD) rats) from the control dose groups of 104-week carcinogenicity studies carried out between 2005 and 2014. The incidence of spontaneous lesions in the bone marrow was higher in mice than in rats, and in both species non-neoplastic lesions were more common than neoplastic lesions. In mice, the most common non-neoplastic lesions in the bone marrow were increased cellularity, pigmented macrophages, and decreased cellularity, and the most common neoplastic lesions were malignant lymphoma, granulocytic leukemia and histiocytic sarcoma. There were occasional sex and site differences (sternum marrow vs femur marrow) in the incidence of a few bone marrow lesions in mice. In rats, the most common non-neoplastic lesions were increased cellularity and stromal fibrosis, and the most common neoplastic lesion was malignant lymphoma. In rats, no sex predilection in the incidence of bone marrow lesions was apparent, and there were no significant site differences in the incidence of lesions. To the best knowledge of the authors, there are no recent reports on spontaneous pathological findings in bone marrow of rodents, and we believe that these results will facilitate the interpretation of background findings and/or their increased incidence in carcinogenicity studies.


Gastro-oesophageal reflux and interstitial lung disease.
Interstitial lung disease is a heterogeneous entity, which encompasses a large number of pulmonary disorders, including idiopathic pulmonary fibrosis. Gastro-oesophageal reflux appears to be more prevalent in idiopathic pulmonary fibrosis than in controls and in patients with secondary pulmonary fibrosis, but its contribution to interstitial lung disease remains unsettled. To prospectively evaluate gastro-oesophageal reflux in patients with interstitial lung disease and to define acid reflux patterns in patients with idiopathic and secondary forms of pulmonary fibrosis. Twenty-eight patients (15M, median age 66 years, interquartile range 60-71) were studied by history taking, pulmonary function tests, high resolution computed tomography, oesophageal manometry and 24h oesophago-gastric pH monitoring. Sixteen patients (57%) had typical reflux symptoms. All patients completed 24h pH metry and 14 underwent oesophageal manometry. An abnormal oesophageal acid exposure was observed in 19 patients (68%), 13 of which complained of predominant typical reflux symptoms. A diagnosis of idiopathic pulmonary fibrosis was made in 18 patients, while the remaining 10 patients were classified as secondary pulmonary fibrosis. Secondary pulmonary fibrosis patients had more restrictive lung defect, as expressed as Tiffeneau index (p<0.05) and greater acid reflux at nighttime (p<0.05) than idiopathic pulmonary fibrosis patients. The prevalence of abnormal acid reflux in interstitial lung disease patients is high and, in particular, patients with secondary pulmonary fibrosis show higher oesophageal acid exposure than those with idiopathic form.


Evaluation of metabolic syndrome related health information on internet in Indian context.
Metabolic Syndrome (MetS) in India is a major contributor to the global increase in CVD. Lifestyle modification programs have been effective in reducing the burden of MetS. The Objective of our study was to evaluate the quality of MetS related health information on the internet in an Indian context. We used a key term "metabolic syndrome" to retrieve websites from Google, Yahoo and Bing search engines by restricting pages from India during May 2012. Previously validated DISCERN tool was used by the three raters to assess 44 websites. All results have been reported as p-values. The most common topics that were covered in these websites included causes and risk factors of MetS (77.27%). On the contrary medication (13.64%), lab tests (11.36%), type of physical activities (6.82%), prognosis and regular check-ups (4.55%) were the least mentioned topics. The website category .org had higher average DISCERN scores as compared to others categories. Limited information was available related to treatment choices, warning signal and informed decision and hence the need exists for further research to develop evidence based health information portal for MetS in an Indian context.


'The biggest worry..': research findings on pain management for Aboriginal peoples in Northern Territory, Australia.
Effective pain management is considered essential during end-of-life care, and is core work for the discipline of hospice and palliative care. However, although there is extensive literature on pain relief during end-of-life care for Caucasians, there are few articles that focus specifically on issues associated with pain management for Australian Aboriginal peoples. In order to address this dearth, the present article provides findings from a National Health and Medical Research Council two-year study on Aboriginal palliative care, conducted in the Northern Territory, that explored and documented issues associated with pain management for rural and remote Aboriginal peoples. The data were collected through open-ended, qualitative interviews conducted with a cross-section of participants (consumers and health professionals) throughout the Northern Territory, Australia. There were a total of seventy-two interviews completed with a wide range of participants including patients (n = 10), carers (n = 19), Aboriginal health care workers (n = 11), healthcare professionals (n = 30) and interpreters (n = 2). The interviews were transcribed verbatim, coded and thematically analysed within a descriptive phenomenological framework. To understand the problems of pain management it is important to appreciate many of the cultural practices and beliefs of Aboriginal peoples. A complexity of cultural relationship rules determine who should and should not be directly involved in providing physical care. Findings from the study show that Aboriginal peoples may have a higher threshold of pain and are less likely to complain - this is particularly so for men, who do not want to appear weak by expressing their pain. Key factors impacting on pain management are cultural concerns about 'blame' and 'pay back'. There is also a fear of Western medicine, stemming from a lack of understanding of clinical notions of pain relief, fear of the administration, side effects and ramifications of medications, and fear that Western pain medications will speed up the dying process and inhibit the passing on of traditional knowledge and secrets that occurs during end-of-life. Strategies posited for ensuring effective pain management include developing trust, timely involvement of the doctor for administering pain medication, provision of emotional support, information giving to decrease fear, provision of the 'right' information to the 'right' person and strengthening of health service provision. The insights provided by a diversity of Aboriginal peoples and the health professionals who care for them provide valuable wisdom with respect to the best way to ensure effective pain management is made available to Australia's first peoples. At the core of this information is the need for cultural sensitivity and respect.


Plants of the genus Allium as antibacterial agents: From tradition to pharmacy.
Plants belonging to the genus Allium are widely cultivated and used all over the world as food and medicinal plants. Since ancient times, these plants, particularly garlic (Allium sativum L.) and onion (Allium cepa L.), have represented important components of typical recipes and traditional healing systems. Not the least of which, their use as food biopreservatives is well documented, due to the relevant antibacterial activity of their extracts and essential oils. In addition to garlic and onion, this review article deals with the main members of the genus Allium, including A. ampeloprasum (Leek), A. schoenoprasum (Chive) and A. ascalonicum (Shallot), focusing both on their ethnonutritional uses and potential as promising food biopreservative agents. Noteworthy, recent research has demonstrated Allium derivatives to be novel components in active edible coatings as well as nanoformulates.


The regulation mechanisms of AhR by molecular chaperone complex.
The AhR, so called the dioxin receptor, is a member of the nuclear receptor superfamily. The ligand-free AhR forms a cytosolic protein complex with the molecular chaperone HSP90, co-chaperone p23, and XAP2 in the cytoplasm. Following ligand binding like 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD), the AhR translocates into the nucleus. Although it has been reported that HSP90 regulates the translocation of the AhR to the nucleus, the precise activation mechanisms of the AhR have not yet been fully understood. AhR consists of the N-terminal bHLH domain containing NLS and NES, the middle PAS domain and the C-terminal transactivation domain. The PAS domain is familiar as a ligand and HSP90 binding domain. In this study, we focused on the bHLH domain that was thought to be a HSP90 binding domain. We investigated the binding properties of bHLH to HSP90. We analyzed the direct interaction of bHLH with HSP90, p23 and XAP2 using purified proteins. We found that not only the PAS domain but also the bHLH domain bound to HSP90. The bHLH domain forms complex with HSP90, p23 and XAP2. We also determined the bHLH binding domain was HSP90 N-domain. The bHLH domain makes a complex with HSP90, p23 and XAP2 via the HSP90 N-domain. Although the NLS is closed in the absence of a ligand, the structure of AhR will be changed in the presence of a ligand, which leads to NLS open, result in the nuclear translocation of AhR.


Conformational mobility of small molecules in glass-forming solutions studied by FTIR spectroscopy.
Small molecules with two or more stable conformations when embedded in a glass-forming liquid (matrix) serve the role of "conformational probes", i.e., their conformational transitions are used to follow local mobility in the matrix. In the present study, conformational probes were embedded in low-molecular-weight glass-forming liquids, and the molecular mobility was studied in a broad temperature range including the glass transition temperature (T(g)). Paraffin oil, dibutylphthalate, bis(2-ethylhexyl)phthalate and isopropylbenzene were used as glass-forming liquids while 1,2-dichloroethane, 1,2-diphenylethane, chlorocyclohexane and bromocyclohexane were used as conformational probes. For some of the matrix/probe systems, the conformational mobility was found to freeze-in at T(g), while for the others it froze-in at certain temperatures T(f)<T(g). In the latter case, it was possible to evaluate the activation energies related to mobility of free volume entities in the glassy matrices. In addition, it was possible to estimate the volumes of sub-molecular groups of the matrix that freeze-in at T(f).


Gene expression profile of adult human bone marrow-derived mesenchymal stem cells stimulated by the chemokine CXCL7.
A variety of chemokines has been shown to recruit human bone marrow-derived mesenchymal stem cells (MSC) and may be potential candidates for chemokine-based tissue regeneration approaches. The aim of our study was to determine whether the chemokine CXCL7 stimulates migration of human bone marrow-derived MSC and to analyze the effect of CXCL7 on the recruitment of MSC on the broad molecular level. Chemotaxis assays documented that high doses of CXCL7 significantly recruited MSC. Gene expression profiling using oligonucleotide microarrays showed that MSC treated with CXCL7 differentially expressed genes related to cell migration, cell adhesion and extracellular matrix remodeling. Pathway analysis showed that CXCL7 induced the expression of all chemokines binding the interleukin (IL) receptors A and B, CXCR1 and CXCR2, as well as the IL6 signal transducer (gp130) and its ligands IL6 and leukemia inhibitory factor (LIF). Induction of differentially expressed chemokines CXCL1-3, CXCL5, and CXCL6 as well as LIF and gp130 in MSC by CXCL7 was verified by real-time polymerase chain reaction. Immunoassay of cell culture supernatants confirmed elevated levels of the interleukins 6 and 8 in MSC upon treatment with CXCL7. Chemotaxis assays showed that interleukin 6 did not recruit MSC. In conclusion, CXCL7 significantly stimulates the migration of human MSC in vitro. Pathway analysis suggests that recruitment of human MSC by CXCL7 is supported by the induction of ligands of the interleukin 8 receptors, synergistically activating the respective signaling pathways.


Resveratrol Activated Sonic Hedgehog Signaling to Enhance Viability of NIH3T3 Cells in Vitro via Regulation of Sirt1.
Injuries of the brain and spinal cord result in the formation of glial (reactive gliosis) and fibrotic (formed by fibroblasts) scars. Recent studies have shown that the fibrotic scar was much more important for hindering regeneration after brain or spinal cord injury than the astrocytic scar. However, it has been given much less attention for effects and mechanism of fibroblasts during formation of the fibrotic scar. Resveratrol may be a potential anti-scarring agent in burn-related scarring and keloid fibroblasts. However, it is unclear whether and how resveratrol affects formation of the fibrotic scar after brain or spinal cord injury. Earlier studies have shown that the activated Shh signaling has anti-apoptosis, anti-oxidation, anti-inflammation properties. Moreover, resveratrol can activate the Shh signaling. However, it is unclear how resveratrol activates the Shh signaling. Resveratrol is a activator of Sirt1. It is unknown whether resveratrol activates the Shh signaling via Sirt1. NIH3T3 cells, a fibroblast cell line, were used as model cells and treated with drugs. Cell viability was assessed by Cell Counting Kit 8. The expressions and activity of Shh signaling pathway proteins were evaluated by immunocytochemistry and Western blotting. Transcriptional activity of Gli-1 was detected with Dual-Luciferase Reporter Gene Assay Kit. Resveratrol, Sirt1 agonist STR1720 and recombinant mouse Shh protein, an activator of hedgehog signaling, enhanced the viability of NIH3T3 cells, promoted Smo to translocated to the primary cilia and Gli-1 entered into the nuclei from cytoplasm, and upregulated expressions of Shh, Ptc-1, Smo, and Gli-1 proteins, which can be reversed by Smo antagonist cyclopamine and Sirt1 antagonist Sirtinol. Additionally, resveratrol increased transcriptional activity of Gli-1. We indicate in the first time that it may be mediated by Sirt1 for resveratrol activating the Shh signaling to enhance viability of NIH3T3 cells, and Sirt1 may be a regulator for upstream of the Shh signaling pathway.This study provides a basis for further investigating effects and mechanism of resveratrol during the formation of fibrous scar after brain or spinal cord injury.


Bimetal Prussian Blue as a Continuously Variable Platform for Investigating the Composition-Activity Relationship of Phosphides-Based Electrocatalysts for Water Oxidation.
Doping unary transition-metal phosphides (TMPs) by secondary metal is a powerful method to improve their catalytic activity for electrochemical oxygen evolution reaction (OER). However, the composition-activity relationship of such doping has not been systematically investigated yet because of the challenge in constructing bimetal TMPs with continuously variable composition while keeping homogenous elemental distribution. Herein, we develop a strategy of using bimetal Prussian blue analogues with homogenous elemental distribution at molecular scale as an ideal platform to achieve bimetal cobalt-iron phosphides (Co1- xFe xP) with a continuously changeable Co/Fe ratio (0 < x < 1) and uniform metal distribution. Such a system allows us to draw out a composition-activity volcano profile of Co1- xFe xP for OER. By optimizing the composition, the best catalytic activity is obtained at the Co/Fe ratio of 1.63 in Co1- xFe xP with small overpotentials of 230 and 268 mV at 10 and 100 mA cm-2, respectively, which outperform most of the reported TMPs. These results may inspire the use of multicomponent molecular platforms to understand composition-dependent performance and explore highly efficient catalysts for diverse applications.


Effect of sleep deprivation on the EEG of learning-impaired children with absence seizures.
Learning-impaired children can present with a clinical history suggesting additional problems, including attention deficit disorder and absence seizures. Physicians use clinical judgment in diagnosing the former, but rely heavily on the electroencephalogram (EEG) to diagnose the latter. Most clinicians are reluctant to treat patients with anticonvulsants without seeing 3 Hz spike-and-wave discharges in an EEG. We studied 21 learning-impaired and 7 control children, ages 5 through 12 years, with normal baseline EEGs. All had an additional EEG recorded after 24 hour sleep deprivation. Nine learning-impaired children had a history compatible with absence seizures; 8 of them showed spike-and-wave discharges after sleep deprivation. Of the 12 learning-impaired children with no history of absence seizures, 2 showed spike-and-wave discharges after sleep deprivation. These results are significant (p less than 0.001). Of the 7 controls, none had epileptiform activity following sleep deprivation. Our results show that routine EEGs may be falsely negative in patients with absence seizures, and that sleep deprivation is a potent activator.


Rapid speciation and determination of vanadium compounds using ion-pair reversed-phase ultra-high-performance liquid chromatography inductively coupled plasma-sector field mass spectrometry.
Environmental vanadium contamination is a potential concern to public health, as evidenced by its place on the U.S. Environmental Protection Agency Drinking Water Contaminant Candidate List as a priority contaminant. Vanadium toxicity varies significantly between different oxidation states; therefore, it is crucial to be able to monitor the speciation of vanadium in environmental samples. In this study, a novel method is described that utilizes ion-pair reversed-phase ultra-high-performance liquid chromatography with inductively coupled plasma-sector field mass spectrometry (IP-RP-UHPLC-ICP-SFMS) to separate vanadyl and vanadate ions and resolve a major polyatomic spectral interference ((35)Cl(16)O(+)) in less than a minute. Detection limits were obtained in the low ngL(-1) (part per trillion) range with linear calibrations across several orders of magnitude (50ngL(-1)-100μgL(-1)). The mechanism of chromatographic retention was elucidated through investigation of the role of ethylenediaminetetraacetic acid, tetrabutylammonium ion and pH on elution. The optimized method was then applied to the speciation of vanadium in local lake water samples.


Nonsteroidal antagonists of the mineralocorticoid receptor.
The broad clinical use of steroidal mineralocorticoid receptor antagonists (MRAs) is limited by the potential risk of inducing hyperkalemia when given on top of renin-angiotensin system blockade. Drug discovery campaigns have been launched aiming for the identification of nonsteroidal MRAs with an improved safety profile. This review analyses the evidence for the potential of improved safety profiles of nonsteroidal MRAs and the current landscape of clinical trials with nonsteroidal MRAs. At least three novel nonsteroidal MRAs have reportedly demonstrated an improved therapeutic index (i.e. less risk for hyperkalemia) in comparison to steroidal antagonists in preclinical models. Five pharmaceutical companies have nonsteroidal MRAs in clinical development with a clear focus on the treatment of chronic kidney diseases. No clinical data have been published so far for MT-3995 (Mitsubishi), SC-3150 (Daiichi-Sankyo), LY2623091 (Eli Lilly) and PF-03882845 (Pfizer). In contrast, data from two clinical phase II trials are available for finerenone (Bayer) which demonstrated safety and efficacy in patients with heart failure and additional chronic kidney diseases, and significantly reduced albuminuria in patients with diabetic nephropathy. Neither hyperkalemia nor reductions in kidney function were limiting factors to its use. Novel, nonsteroidal MRAs are currently tested in clinical trials. Based on preclinical and first clinical data, these nonsteroidal MRAs might overcome the limitations of today's steroidal antagonists.


Identification of basophilic cells that express mast cell granule proteases in the peripheral blood of asthma, allergy, and drug-reactive patients.
Metachromatic cells in the peripheral blood of patients with asthma, allergy, or an allergic drug reaction were evaluated for their nuclear morphology, surface expression of the mast cell (MC) marker c-kit, surface expression of the basophil marker Bsp-1, and granule expression of MC proteases. Consistent with previous findings by others, Bsp-1+/metachromatic cells represented <1% of the cells in the peripheral blood of normal individuals. These cells generally contained segmented nuclei. Very little, if any, tryptase (Try), chymase (Chy), or carboxypeptidase A (CPA) was found in their granules, and very little, if any, c-kit was observed on their surfaces. The number of metachromatic cells increased in the peripheral blood of the three groups of patients. Like the basophils in normal individuals, most of these metachromatic cells contained segmented nuclei and expressed Bsp-1. However, in contrast to the basophils in normal individuals, many of the metachromatic cells in the three patient groups expressed c-kit, Try, Chy, and/or CPA. That the metachromatic cells in the blood of our patients have some features of MCs and some features of basophils suggests that human basophils and MCs are derived from a common progenitor. As assessed by the chloroacetate esterase cytochemical assay, the immunoreactive Chy in the peripheral blood of these patients is enzymatically active. Because MC proteases regulate numerous immunologic and other biologic systems, the expression of Try, Chy, and/or CPA in a peripheral blood-localized cell in an individual having asthma, allergy, or an allergic drug reaction has important clinical implications.


Atlantoaxial Fusion: Sixteen Years of Epidemiology, Indications, and Complications in New York State.
A retrospective state database analysis. The aim of this study was to describe the epidemiology and complications of as well as indications for primary isolated atlantoaxial fusion. Atlantoaxial fusion involves unique indications, techniques, and complications. There is limited epidemiologic literature focused specifically on this procedure. New York's Statewide Planning and Research Cooperative System database, an all-payer hospitalization reporting system, was queried to identify all patients undergoing primary isolated atlantoaxial fusion in the state from 1997 to 2012. Demographic and clinical data were extracted and analyzed. United States Census Bureau figures were used to calculate population-adjusted surgical rates. One thousand five hundred fifty-nine patients underwent isolated primary atlantoaxial fusion during the study period. The overall population-adjusted annual surgical rate did not change significantly over time. By 2012, individuals aged ≥70 years had the highest incidence of surgery [2.37 per 100,000 population; 95% confidence interval (95% CI) 1.68-3.07]. Medicare was the most common payer (44.0% of claims). Approximately 85% of patients had a Charlson/Deyo Comorbidity Index of zero or one. Over time, a significantly lower proportion of atlantoaxial fusions were attributable to rheumatic disease, and a significantly higher proportion were due to fracture. By 2012, management of fractures was the most common indication for C1-C2 fusion (44.1% of cases). Dysphagia or dysphonia occurred after 0.8% of cases, dural tear after 0.3%, infection after 0.5%, and seroma, hematoma, or hemorrhage after 0.5%. In-hospital mortality was 2.7%, of which 76% had fracture as the surgical indication. Isolated atlantoaxial fusions have been performed at a stable, low level over the past 16 years in New York. Although most of these patients are relatively healthy pre-operatively, approximately one in 10 experience an in-hospital complication and nearly 3% die in-hospital. Knowledge of these risks will hopefully spur further efforts to minimize them and allow for more accurate counseling of patients and their families. 4.


Susceptibility of pregnant women to toxoplasma infection--potential benefits for newborn screening.
Congenital toxoplasmosis (CT) arises as a result of new acquisition of Toxoplasma infection by a susceptible woman during pregnancy. Early detection of CT through neonatal screening programmes could optimize management and improve infant outcome. This study sought to estimate the prevalence of Toxoplasma susceptibility in pregnant women. As detection of Toxoplasma antibodies in neonatal blood reflects maternal exposure history, maternal antibody seroprevalence was determined using anonymized residual blood from newborn screening cards. A total of 20,252 cards were tested in 1 year. 4,991 (24.6%) cards tested positive for Toxoplasma antibody. Results were stratified by county. Toxoplasma antibody seroprevalence rates of 25% indicated that Toxoplasma infection is common in Ireland and that up to 75% of women remain susceptible to primary infection during pregnancy. This study aimed to a) determine the seroprevalence of Toxoplasma antibody in pregnant women, and hence b) estimate the risk for acquisition of primary toxoplasmosis in pregnancy in order to support an application to fund a pilot newborn screening programme.


Eimeria tenella heat shock protein 70 enhances protection of recombinant microneme protein MIC2 subunit antigen vaccination against E. tenella challenge.
Heat shock proteins have been reported to stimulate the immune system via innate receptors. Our study found that the novel immunopotentiator, Eimeria tenella (E. tenella) heat shock protein 70 (HSP70), enhanced protective immunity elicited by E. tenella antigen microneme protein 2 (EtMIC2) against avian coccidiosis. It demonstrated that the expression of TLR2 and TLR4 were strongly upregulated in EtHSP70 and EtMIC2 plus EtHSP70 stimulated chicken embryo fibroblasts (CEF) compared with untreated controls and EtMIC2 alone. In addition, the same treatment induced high levels of interleukin (IL)-12 and interferon (IFN)-γ that are critical cytokines of innate immunity. In vivo experiments involved using broiler chickens subcutaneously immunized with EtMIC2 alone or EtMIC2 plus EtHSP70 at 7 and 14 days post-hatch, which were then orally challenged with live E. tenella at 7 days following secondary immunization. Body weight gains, cecal lesion scores, fecal oocyst shedding, serum antibody responses against MIC2, and intestinal cytokine transcript levels were assessed as measures of protective immunity. Chickens immunized with EtMIC2 plus EtHSP70 showed increased body weight gains, decreased oocyst shedding, increased serum antibody responses, and high levels of IL-12, IFN-γ, and IL-17 compared with the EtMIC2 only or control groups. Moreover, chickens immunized with EtHSP70 alone showed significantly protective effect against E. tenella infection. In summary, this study provides the first evidence of the immunoenhancing activities of EtHSP70 in poultry.


The lipopolysaccharide polyclonal response is a function of the background response.
The aims of this report are: to investigate the kinetics of the E. coli lipopolysaccharide polyclonal response as a function of the background response, to test the capacity of lipopolysaccharide to activate presumably resting memory cells for IgM. Because mice do not produce a plaque-forming cell background against rat erythrocytes, the mouse-rat erythrocyte system was chosen. Random bred Swiss albino mice, aged 2-3 months, were inoculated with rat erythrocytes (5 X 10(5)/mouse) and the course of the plaque response was followed up; no response was detected on day 0, it rose sharply up to day 4, dropped abruptly between days 4 and 7 and trailed off afterwards to disappear by day 29. As to the lipopolysaccharide-triggered anti-rat erythrocyte response, lipopolysaccharide (100 micrograms/mouse) was inoculated on days--3, 1, 4, 7, 18 and 29. post-rat erythrocytes and the responses were measured 3 days after lipopolysaccharide inoculation. The curve paralleled that of the antigenic response, although at a higher level; thus, both the specific and the polyclonal responses followed the same kinetics. At one month post-rat erythrocyte priming, the animals displayed a background comparable to that of naive mice, although their IgM response to a booster (2 X 10(7) erythrocytes, iv) was faster and greater (p less than 0.01) than the primary response to the same dose; i.e., there was IgM memory. Nevertheless, lipopolysaccharide stimulation (100 micrograms/mouse, ip) of primed animals gave a response comparable (p greater than 0.10) to that of naive mice; i.e., lipopolysaccharide alone was unable to elicit a secondary-like response.


Co-expression of membrane-bound TNF-alpha type 1 and 2 receptors differ in the subsets of immunocompetent cells.
The immunoregulatory cytokine tumor necrosis factor α plays crucial roles in the pathogenesis of a broad spectrum of disorders. However, its effect may depend on the expression and co-expression of receptors on the target cell. The aim of the present study was to evaluate the expression levels of type 1 and 2 tumor necrosis factor α receptors (TNFR1/2) on individual cell subsets from peripheral blood of healthy volunteers. Flow cytometry analysis was used to study whole populations as well as subsets (T regulatory cells, T memory cells, cytotoxic T cells, T helper cells). Significant differences in the co-expression of TNFR1/2 were seen within subsets and total pools. Further studies are necessary to explore the implications of the observed differences in the modulation of tumor necrosis factor α function in health and pathology.


Syndactyly of the toes.
The experience gained through the management of 43 patients with syndactyly of the toes is presented. The incidence appears to be similar to that of syndactyly of the fingers. Type 1 syndactyly, or zygodactyly, always presented itself as a cosmetic problem; its correction is occasionally indicated and the procedure used is discussed. Type 2 syndactyly, or polysyndactyly, represents a functional problem and deserves surgical correction. My negative experience with the more complex procedures described for the correction of polysyndactyly is presented as well as my satisfaction with the simpler procedures. Suggestions for management are offered.


Rhizopus stolonifer mediated biosynthesis of biocompatible cadmium chalcogenide quantum dots.
We report an efficient method to biosynthesize biocompatible cadmium telluride and cadmium sulphide quantum dots from the fungus Rhizopus stolonifer. The suspension of the quantum dots exhibited purple and greenish-blue luminescence respectively upon UV light illumination. Photoluminescence spectroscopy, X-ray diffraction, and transmission electron microscopy confirms the formation of the quantum dots. From the photoluminescence spectrum the emission maxima is found to be 424 and 476nm respectively. The X-ray diffraction of the quantum dots matches with results reported in literature. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay for cell viability evaluation carried out on 3-days transfer, inoculum 3×105 cells, embryonic fibroblast cells lines shows that more than 80% of the cells are viable even after 48h, indicating the biocompatible nature of the quantum dots. A good contrast in imaging has been obtained upon incorporating the quantum dots in human breast adenocarcinoma Michigan Cancer Foundation-7 cell lines.


[Incidence of unplanned extubations in a neonatal intensive care unit. A before and after study].
Unplanned extubations (UE) of mechanically ventilated newborns involves an undesirable increase in morbidity and mortality. A 2-stage study compared the frequency of UE in a Neonatal Intensive Care Unit before and after the implementation of a program of preventive measures to decrease UE. A before and after prospective study included all mechanically ventilated newborns participating in the 2 stage study from May-December 2011 and June-December 2012. In stage 1, the rate of UE per 100 intubated patient days was calculated and the characteristics of unplanned extubated newborns, circumstances of UE occurrence and need for re-intubation were studied. Consequently, a program of preventive measures for UE was designed and implemented, with the same variables being analysed in stage 2. No differences were found in patient characteristics during the two stages. Stage 1, incidence of UE was 5/100 intubated patient days; Stage 2, 4.5 UE/100 intubated patient days (P=.657). In both stages, most UE occurred during patient handling with re-intubation incidence at 77.4% and 67.7%, respectively. The combined rate of both stages during summer months of July, August and September was 6.2 UE/100 intubation days, in contrast with the remaining months of both stages: UE incidence rate, 3.4 UE/100 intubation days (p=.043). The implementation of a preventive measures program did not significantly reduce the incidence of UE. The summer period showed the highest incidence of UE.


[Migration of an intrauterine device to the bladder].
The authors report the case of a patient presenting with recurrent urinary tract infection and deterioration of the general status. Ultrasonography, plain x-rays and especially cystoscopy demonstrated the presence in the bladder of an intrauterine contraceptive device inserted 4 years previously. Endoscopic removal combined with antibiotics ensured cure of this patient.


Influence of age, gender, health status, and depression on quantitative EEG.
Quantitative electroencephalography (QEEG) has shown increasing utility in assessing brain function in clinical research studies of depression. QEEG findings may be influenced by a variety of factors other than the presence of depression, including age, gender, depression severity, and physical health status. Many of these factors have not been systematically evaluated. We therefore examined QEEG measures in 104 subjects with depression and normal controls to determine the influence of these factors. We examined QEEG power as well as cordance, a QEEG measure that has a stronger association with cerebral perfusion than conventional QEEG measures. Prefrontal cordance in the theta band has been associated with the pathophysiology of depression and response to treatment. We found that prefrontal cordance and relative power in the theta band were unaffected by age, gender, severity of depression, and health status, while prefrontal absolute power was higher in women than men. All of these measures were different from global measures of absolute and relative power, which were influenced by age, gender, and health status. These findings suggest that prefrontal cordance in depressed patients is not significantly affected by factors of age, gender, severity of depression, or physical illness. Global measures of power, and to a lesser extent prefrontal absolute power, must be interpreted with regard to confounding factors of age, gender, physical illness, and severity of depression.


The rate of appearance of thyroid hormone nuclear receptor is increased during deoxyribose nucleic acid synthesis in GC cells: analysis of thymidine-treated GC cells using dense amino acid labeling.
The growth rate of GH-producing rat pituitary tumor cells (GC cells) is dependent on thyroid hormone. Previous studies have shown that GC cells can be partially synchronized in the DNA synthesis phase (S-phase) of the cell cycle by a 25-h incubation with 2 mM thymidine. Measurements in GC cells partially synchronized in S-phase showed a significant increase in cellular thyroid hormone nuclear receptor appearance and disappearance in thymidine-treated GC cells using dense amino acid labeling and subsequent sucrose gradient ultracentrifugation. In comparison with GC cells in asynchronous culture, thymidine treatment causes a steady state increase in thyroid hormone nuclear receptors which results from an increase in the receptor appearance rate without an effect on the disappearance rate. Thus, position in the cell cycle appears to be another factor that influences the appearance rate of nuclear thyroid hormone receptors.


Emil Kraepelin and modern psychiatry.
A brief account is given of the career and scientific contributions of the German psychiatrist, Emil Kraepelin, to whom the concept of schizophrenia is generally attributed. Attention is also drawn to his social and political outlook and its relationship to his work. The implications for contemporary psychiatry are outlined.


Fast spin rotations by optically controlled geometric phases in a charge-tunable InAs quantum dot.
We demonstrate optical control of the geometric phase acquired by one of the spin states of an electron confined in a charge-tunable InAs quantum dot via cyclic 2pi excitations of an optical transition in the dot. In the presence of a constant in-plane magnetic field, these optically induced geometric phases result in the effective rotation of the spin about the magnetic field axis and manifest as phase shifts in the spin quantum beat signal generated by two time-delayed circularly polarized optical pulses. The geometric phases generated in this manner more generally perform the role of a spin phase gate, proving potentially useful for quantum information applications.


[The duration of the storage of the electrical characteristics of command neurons in the acquisition of long-term sensitization in the snail].
The retention of the long-term sensitization (LTS) of defensive reflex and dynamics of change in electric characteristics (membrane potential (Vm) and action potential generation threshold (Vt)) of command neurons of defensive reflex was studied in a snail during behavioral tests. The membrane mechanisms were analyzed by measuring electrical characteristics of the LPa3, RPa3, LPa2, and RPa2 command neurons on the 1st, 4th, 7th, 10th, and 14th days after the LTS formation and 1 month later. The membrane potential and threshold potential in sensitized snails (-54.1 +/- 2.0 and 24.5 +/- 1.4 microV, respectively) were significantly (p < 0.001) decreased in comparison with the control animals (-60.9 +/- 0.8 and 19.9 +/- 0.6 microV respectively). These changes retained within 14 days after the LTS formation. The results suggest the long-term retention of the increased excitability of command neurons. A month after the LTS formation, the duration of the defensive reflex returned to the initial level and the electric characteristics of command neurons did not significantly differ from the control (-61.1 +/- 2.0 and 19.3 +/- 1.4 microV, respectively).


Use of atropine in patients with acute myocardial infarction and sinus bradycardia.
Fifty-six patients with acute myocardial infarction complicated by sinus bradycardia (SB) were treated with intravenous atropine and monitored in a coronary care unit. Atropine decreased or completely abolished premature ventricular contractions (PVCs) and/or bouts of accelerated idioventricular rhythm in 27 of 31 patients (87%) and brought systemic blood pressure up to normal in 15 of 17 patients (88%) with hypotension. In addition, atropine administration was associated with improved atrioventricular conduction in 11 of 13 patients (85%) with acute inferior myocardial infarction associated with 2 degrees or 3 degrees atrioventricular block. Seven patients developed ten significant adverse effects: ventricular tachycardia or fibrillation in three, sustainedsinus tachycardia in three, increased PVCs in three, and toxic psychosis in one. These major adverse effects correlated with either a higher initial dose of atropine (i.e., 1.0 mg aa compared with the usual 0.5 or 0.6 mg) or a total cumulative dose exceeding 2.5 mg over 21/2 hours. Atropine is the drug of choice for management of patients with SB and hypotension and is effective in the treatment of ventricular arrhythmias as well as conduction disturbances in patients with inferior myocardial infarction. Serious adverse effects, however, preclude use of atropine without careful medical supervision.


Transplantation of filgrastim-mobilized peripheral blood stem cells from HLA-identical sibling or alternative family donors in patients with hematologic malignancies: a prospective comparison on clinical outcome, immune reconstitution, and hematopoietic chimerism.
The clinical results, cellular immune reconstitution, and hematopoietic chimerism obtained after transplantation of recombinant human granulocyte colony-stimulating factor mobilized allogeneic peripheral blood stem cells (PBSCs) from genotypically human leukocyte antigen (HLA)-identical sibling (n = 36) or alternative family donors (n = 24) were prospectively compared in patients with hematologic malignancies. Thirty-two of 34 evaluable patients with HLA-identical sibling donors and all patients with alternative family donors achieved trilineage engraftment. The median time intervals to reach peripheral neutrophil counts <500/microL (13 v 17 days) or <1,000/microL (16 v 19 days) and unsupported platelet counts <20,000/microL (11 v 15 days) or <50, 000/microL (19 v 24 days) as well as red blood cell and platelet transfusion requirements were not significantly different between both patient subsets. The cumulative probability of grades II through IV acute graft-versus-host disease (GVHD) for the 60 study patients was 48% +/- 10% but ranged between 86% +/- 12% in patients whose donors had at least one HLA-A,B,DR,DQ,DP antigen disparity in direction to acute GVHD, and 25% +/- 9% in recipients of GVHD-matched transplants (P < .003). The 2-year survival estimates were 54% +/- 10% for patients with alternative family donors and 65% +/- 9% for patients with HLA-identical sibling donors. Multivariate analysis identified the pretransplantation disease stage, patient age, and acute GVHD as independent predictors of overall and disease-free survival, whereas alternative family donors alone had no adverse effect on these clinical endpoints. Monthly monitoring of peripheral blood T-helper cell subsets, B cells, and monocytes during the first year posttransplantation showed a nearly identical course of immune cell reconstitution in both patient subsets. In addition, no differences in the proportions of complete chimeric patients were detectable between the two patient subsets by sex chromosome and variable number of tandem repeats analysis up to 12 months posttransplantation. In conclusion, PBSCs from alternative family donors represent an attractive source for allogeneic transplantation in patients lacking HLA-identical sibling donors and should be further evaluated in comparison with marrow transplants from alternative family donors.


Bimanual coordination: constraints imposed by the relative timing of homologous muscle activation.
It has often been supposed that patterns of rhythmic bimanual coordination in which homologous muscles are engaged simultaneously, are performed in a more stable manner than those in which the same muscles are activated in an alternating fashion. In order to assess the efficacy of this constraint, the present study investigated the effect of forearm posture (prone or supine) on bimanual abduction-adduction movements of the wrist in isodirectional and non-isodirectional modes of coordination. Irrespective of forearm posture, non-isodirectional coordination was observed to be more stable than isodirectional coordination. In the latter condition, there was a more severe deterioration of coordination accuracy/stability as a function of cycling frequency than in the former condition. With elevations in cycling frequency, the performers recruited extra mechanical degrees of freedom, principally via flexion-extension of the wrist, which gave rise to increasing motion in the vertical plane. The increases in movement amplitude in the vertical plane were accompanied by decreasing amplitude in the horizontal plane. In agreement with previous studies, the present findings confirm that the relative timing of homologous muscle activation acts as a principal constraint upon the stability of interlimb coordination. Furthermore, it is argued that the use of manipulations of limb posture to investigate the role of other classes of constraint (e.g. perceptual) should be approached with caution because such manipulations affect the mapping between muscle activation patterns, movement dynamics and kinematics.


Lorraine Anne Allan.
As an advocate for animals and students, she was a generous with her time in helping vet nurses to learn and develop. She will be remembered as an inspirational teacher.


A reference genome for pea provides insight into legume genome evolution.
We report the first annotated chromosome-level reference genome assembly for pea, Gregor Mendel's original genetic model. Phylogenetics and paleogenomics show genomic rearrangements across legumes and suggest a major role for repetitive elements in pea genome evolution. Compared to other sequenced Leguminosae genomes, the pea genome shows intense gene dynamics, most likely associated with genome size expansion when the Fabeae diverged from its sister tribes. During Pisum evolution, translocation and transposition differentially occurred across lineages. This reference sequence will accelerate our understanding of the molecular basis of agronomically important traits and support crop improvement.


Analysis of a shared pancreatic cancer antigen recognized by an HLA-A*2601-restricted cytotoxic T-lymphocyte clone.
We have generated HLA-A*2601-restricted CD8+ CTL clones against an autologous pancreatic cancer cell line. To characterize the antigen expressed on the cancer cells. We assessed cytotoxic activities and cytokine production of these CTL clones reacting against cancer cell lines that stably or transiently expressed the HLA-A*2601 gene. These CTL clones recognized 4 of 10 allogeneic pancreatic cancer cell lines and a gallbladder cancer cell line in the context of HLA-A*2601. However, the CTL clones did not recognize three hepatocellular carcinoma cell lines, two esophageal squamous cell carcinoma cell lines, or a lung adenocarcinoma cell line. Thus, the CTL clones may recognize a shared, but not ubiquitously expressed, tumor antigen on pancreatic and gallbladder cancer cells.


Rhomboid flap.
The rhomboid flap is a practical and efficient method of repairing small- to moderate-size surface defects of the head and neck. This flap has the advantage of combining the lateral shift and rotational methods of transfer that aid in the closure of the donor site as the flap is transposed. A description of this flap, the principle of its design, its clinical application, and potential pitfalls are presented.


Precision Medicine and the Changing Landscape of Research Ethics.
President Barack Obama announced the launch of the National Institutes of Health Precision Medicine Initiative® (PMI) in January 2015. Precision medicine includes the concept of individualized or personalized medicine at a more exact level through advances in science and technology, such as genetics and genomics sequencing. Although many disease processes will be investigated through the precision medicine lens for greater understanding and improved treatment responses, oncology research and translation to practice is leading the initiative's debut, referred to as the near-term focus.


Proteomic analysis of lamellar bodies isolated from amniotic fluid: implications for function.
Lamellar body count (LBC) in amniotic fluid is a well-established method for assessing fetal lung maturity. However, the biogenesis and function of lamellar bodies (LBs) secreted into the amniotic fluid have not been formally assessed. We purified LBs from amniotic fluids obtained from term gestation pregnancies that had been determined to have mature LBC. Using tandem mass spectrometry, we identified 122 unique proteins in the LB preparations from the amniotic fluids. There was minimal overlap between the proteins identified in amniotic fluid LB and those reported for human epidermis LB. In contrast, there was >40% concordance with the proteome of rat lung LBs despite species differences. Classification of the identified proteins into functional bins demonstrated that the preponderance of amniotic fluid LB proteins was associated with host defense or anti-inflammatory functions. These data suggest that amniotic fluid LBs are derived from lung secretions and may play an important role in innate host defense of the fetus.


Supranutritional vitamin E supplementation in pigs: Influence on subcellular deposition of α-tocopherol and on oxidative stability by conventional and derivative spectrophotometry.
The influence of three levels of vitamin E (30, 200 and 1000mg kg(-1)) in the diet of pigs on the subcellular deposition of α-tocopherol in muscle and on the oxidative stability determined by conventional and first derivative Spectrophotometry was studied. The content of α-tocopherol in m. gluteo biceps and in mitochondrial and microsomal fractions of the muscle significantly increased (p < 0.01) with increasing levels of dietary vitamin E. Concentrations of α-tocopherol in muscle, mitochondria and microsomes of pigs fed diet supplemented with 1000 mg kg(-1) α-tocopheryl acetate were 3.2-, 6.1- and 5.6-fold greater, respectively, than those in their counterparts from the control animals. These differences in α-tocopherol concentration in the subcellular fractions and intact muscle resulted in enhanced stability of the membranes and the tissue when exposed to iron-ascorbate induced peroxidation. When lipid oxidation in the same samples was further measured by the first derivative method, the resultant MDA-TBA values were 59-69% lower in tissue samples, 16-19% lower in mitochondria and 6-9% lower in microsomes than the conventional TBARS values.


Characteristics of optical coherence tomographic hyperreflective foci in retinal vein occlusion.
To evaluate the hyperreflective foci in branch retinal vein occlusion and central retinal vein occlusion depicted by spectral-domain optical coherence tomography (OCT). Consecutive series of 73 eyes of 73 patients with retinal vein occlusion (58 branch retinal vein occlusion and 15 central retinal vein occlusion) who had Spectralis OCT images obtained were retrospectively reviewed, comparing color fundus photographs and fluorescein angiography. Hyperreflective foci were detected in 54 eyes (74.0%) by spectral-domain OCT, and hard exudates were detected in 17 eyes (23.3%) by color fundus photography. Hard exudates on the color photographs corresponded to the confluence of hyperreflective foci mainly around the outer plexiform layer in the unaffected areas on the spectral-domain OCT images, whereas fine hyperreflective foci were scattered in all retinal layers of the affected areas (P < 0.001). Most eyes with hyperreflective foci attached to the inner side of the external limiting membrane also had serous retinal detachment (P < 0.001). Compared with diabetic macular edema, we did not find subfoveal hard exudates in retinal vein occlusion. Hyperreflective foci delineated on spectral-domain OCT suggest the pathogenesis regarding the flow of extravasated blood constituents in retinal vein occlusion.


Control of disinfection by-product formation using ozone-based advanced oxidation processes.
The effects of ozone dosage, water temperature and catalyst addition in an ozonation-fluidized bed reactor (O3/FBR) on treated water quality and on the control of chlorinated and ozonated disinfection by-products (DBPs) were investigated. A biofiltration column was used to evaluate its removal efficiency on biodegradable organic matter and to reduce DBP formation. The Dong-Gang River, polluted by agricultural and domestic wastewater in Pingtung, Taiwan, was used as the water source. The treated water quality in terms of dissolved organic carbon (DOC), biodegradable DOC, ultraviolet absorbance at 254 nm (UV254) and specific UV absorbance (SUVA) improved with increasing ozone and catalyst dosages. Catalytic ozonation was more effective than ozonation alone at reducing the formation of DBPs at a given dosage. Experimental results show that water temperature had little effect on the treated water quality with the O3/FBR system used in this study (p > 0.05). The combination of O3/FBR and the biofiltration process effectively decreased the amount ofDBP precursors. The concentration of total trihalomethanes (TTHMs) was less than the maximum contaminant level (MCL) requirement, which is 80 microg/L, for all treated waters and the concentration of five haloacetic acids (HAA5) fell below 60 microg/L with an ozone dosage higher than 2.5 mg/L.


Classical statistical considerations in medical decision models.
The influence of sampling error on decision-analytic models was investigated to determine how these errors affect model reliability. Formulas were developed to relate statistical error in the probability decision threshold and gain in expected utility to the error in the data samples upon which such models are based. The formulas were validated in a simulation experiment and then applied to a hypothetical decision model and to the clinical problem of immediate surgery versus continued observation in suspected acute appendicitis. The results of this analysis show that modest statistical error affecting any variable in a decision model may be amplified into a substantially larger error in both the probability decision threshold and the gain in utility predicted by the model. In addition, when errors are present simultaneously in several variables, they may compound to unexpectedly large magnitudes, rendering the model unreliable over a wide range of disease probability. The interpretation of the results of a decision analysis should be viewed along a continuum that takes into account both the magnitude of the gain or loss in expected utility predicted by the model and a quantitative measure of the reliability of this prediction. Whenever possible, a determination of statistical error should be an integral part of any formal decision analysis.


Venom of the crotaline snake Atropoides nummifer (jumping viper) from Guatemala and Honduras: comparative toxicological characterization, isolation of a myotoxic phospholipase A(2) homologue and neutralization by two antivenoms.
A comparative study was performed on the venoms of the crotaline snake Atropoides nummifer from Guatemala and Honduras. SDS-polyacrylamide gel electrophoresis, under reducing conditions, revealed a highly similar pattern of these venoms, and between them and the venom of the same species from Costa Rica. Similar patterns were also observed in ion-exchange chromatography on CM-Shephadex C-25, in which a highly basic myotoxic fraction was present. This fraction was devoid of phospholipase A(2) activity and strongly reacted, by enzyme-immunoassay, with an antiserum against Bothrops asper myotoxin II, a Lys-49 phospholipase A(2) homologue. A basic myotoxin of 16 kDa was isolated to homogeneity from the venom of A. nummifer from Honduras, showing amino acid composition and N-terminal sequence similar to those of Lys-49 phospholipase A(2) variants previously isolated from other crotaline snake venoms. Guatemalan and Honduran A. nummifer venoms have a qualitatively similar toxicological profile, characterized by: lethal; hemorrhagic; myotoxic; edema-forming; coagulant; and defibrinating activities, although there were significant quantitative variations in some of these activities between the two venoms. Neutralization of toxic activities by two commercially-available antivenoms in the region was studied. Polyvalent antivenom produced by Instituto Clodomiro Picado was effective in the neutralization of: lethal; hemorrhagic; myotoxic; coagulant; defibrinating; and phospholipase A(2) activities, but ineffective against edema-forming activity. On the other hand, MYN polyvalent antivenom neutralized: hemorrhagic; myotoxic; coagulant; defibrinating; and phospholipase A(2) activities, albeit with a lower potency than Instituto Clodomiro Picado antivenom. MYN antivenom failed to neutralize lethal and edema-forming activities of A. nummifer venoms.


Antibiotics for treating scrub typhus.
Scrub typhus is a cause of fever in regions of Asia and the Pacific. Recently, resistance to antibiotics has been reported. To assess treatment regimens for scrub typhus through time to fever resolution and incidence of relapse. Cochrane Controlled Trials Register, MEDLINE, EMBASE, and contacted individual researchers for unpublished data. Randomised and pseudorandomised studies; patients diagnosed with scrub typhus defined by authors; any comparison of antibiotic regimens for treating scrub typhus. Trial quality was assessed, and data abstracted by both reviewers. Three trials met the inclusion criteria. One small trial compared tetracycline against chloramphenicol. The other two trials compared doxycycline with tetracycline, and showed little difference in the proportion febrile at 48 hours, with no relapses after either drug. Tetracycline and doxycycline seem to be effective in treating scrub typhus. Further research is required to identify appropriate treatment in areas where Orientia tsutsugamushi resistant to doxycycline has been reported.


The cerebrospinal fluid immunoglobulin G index as a diagnostic aid in multiple sclerosis: a Bayesian approach.
Having determined immunoglobulin G (IgG) and albumin concentrations in 1100 cerebrospinal fluid and serum samples, we calculated the IgG index. Likelihood ratios for multiple sclerosis were calculated by using a training set consisting of 100 patients with definite multiple sclerosis and one consisting of 97 patients suffering from diseases from which multiple sclerosis must be differentiated. Predictive values for multiple sclerosis, given different values for the IgG index, are given in a graphical representation of Bayes' theorem. We conclude that this approach increases the diagnostic usefulness of the IgG index for the diagnosis of multiple sclerosis.


[Exocrine function of the pancreas in patients with chronic hepatitis and liver cirrhosis of various etiologies].
Exocrinous performance of the pancreatic gland under secretin-pancreozymin stimulation was studied in 76 patients with chronic diffuse diseases of the liver who were distinguished into 6 groups: those who suffered from chronic persistent hepatitis of viral and alcohol origin, chronic active hepatitis of viral origin, cirrhosis of the liver of viral and alcohol origin, primary biliary hepatocirrhosis. The results obtained were correlated with those from 11 normal persons (controls). Out of 76 examinees the disorders of exocrinous performance of the pancreatic gland were revealed in 75 persons. The most characteristic features were: a decrease in the basal and an increase in the stimulated volume of the pancreatic juice; a reduction of both basal and stimulated production of bicarbonates; a decrease in the trypsin and amylase fasting levels and their increment in the stimulated juice of the pancreatic gland. Disorder in the production of bicarbonates was stated as a most characteristic feature in the patients both with viral and alcohol origin of the disease but it was mostly manifest in the patients with hepatocirrhosis. Pronounced elevation of the activity of amylase and trypsin in the pancreatic juice was observed in patients with very high activity of disease development and in the patients who continuously used large amounts of alcohol. The authors suspected that alcohol abuse and the effect of hepatitis virus had an equal pathogenic impact on the liver and pancreatic gland.


Hemolytic-uremic syndrome associated with pancreatitis in an HIV-positive patient.
Hemolytic-uremic syndrome (HUS) is a newly recognized hematologic manifestation of HIV infection that may be triggered by local or systemic infections as well as by immunological disorders. We report the case of a 36-year-old HIV-positive man, an intravenous drug abuser who developed HUS during an episode of acute pancreatitis. Hematologic and clinical improvement occurred following 2 weeks of nonaggressive therapy including vitamin E and fresh-frozen plasma.


The insular formations of the dolphin brain: quantitative cytoarchitectonic studies of the insular component of the limbic lobe.
The large insula of the bottlenose dolphin consists of radial gyri arising, in fanlike fashion, from the transverse insular gyrus, and is covered completely by the frontal, parietal, and temporal opercula . On cytoarchitectonic grounds, the dolphin insula is divided into anterior, middle, and posterior sectors that may be the equivalent of the three similar sectors present in the primate insula. Rostrocaudally, these sectors become increasingly more homogeneous and less laminated. Within each sector progressive differentiation occurs in the direction of the circular sulcus. A transitional cortex, the peripaleocortex in the transverse insular gyrus, is interposed between the prepiriform and the periamygdalar cortex and the proisocortex of the insula proper. This peripaleocortex consists of outer and inner cellular strata separated by a hypocellular lamina dissecans. The outer cell stratum is continuous with layers II and III of the insular proisocortex ; the more prominent inner stratum is continuous with proisocortical layers V and VI; the intervening lamina dissecans becomes partially filled, mostly with modified pyramidal cells of medium size that may constitute an incipient layer IV. A band of myelinated fibers corresponding to the external band of Baillarger is found within the lamina dissecans. The anterior insular sector is characterized by distinct lamination and a well-defined, ribbonlike layer Va. In the middle sector, the cortex is internodense and lamination is less clear. The posterior sector is even less laminated and tends to be externodense . Within each sector, lamination becomes clearer in the direction of the circular sulcus. Furthermore, the rostrocaudal architectonic changes suggest a possible transition from a motor-type to a sensory-type cortex. Beyond the insula, the architecture of the opercular cortices reflect, in turn, the influences of the insular sectors.


A case of cystic lymphocytic hypophysitis with cacosmia and hypopituitarism.
Lymphocytic hypophysitis is a rare inflammatory disease of the pituitary gland that is being increasingly recognized as a cause of hypopituitarism. This condition may be due to an autoimmune pituitary destruction which usually occurs in young women during pregnancy or in the immediate postpartum period. We describe a case of cystic pituitary mass in a thirty-eight year-old woman presenting with nausea, vomiting, cold intolerance, blurring of vision and the presence of disagreeable odors for a one-month period. She had secondary amenorrhea and galactorrhea for three months. Combined anterior pituitary stimulation test confirmed the diagnosis of hypopituitarism. Magnetic resonance imaging scan with enhancement showed a huge cystic sellar mass with suprasellar extension and thickening of the pituitary stalk. Transsphenoidal exploration was performed with preoperative diagnosis of pituitary macroadenoma with cystic necrosis. Histological examination revealed lymphocytic hypophysitis characteristic of diffuse, dense lymphocytes and plasma cells infiltration with surrounding interstitial reactive fibrosis. Postoperatively, the patient's olfactory function returned to normal but combined anterior pituitary stimulation test showed persistence of hypopituitarism with mild hyperprolactinemia. Prednisolone, thyroxine and estrogen replacements were started and clinical symptoms were much improved. In summary, we report an extremely rare case of a woman with cystic lymphocytic hypophysitis with cacosmia and hypopituitarism, confirmed by histological examination.


Hormone replacement therapy, insulin sensitivity, and abdominal obesity in postmenopausal women.
The purpose of this study was to determine whether insulin sensitivity differs between postmenopausal women taking estradiol, women on estrogen plus progesterone hormone replacement therapy (HRT), and women not on HRT and whether differences are explained by the differences in total and/or abdominal adiposity and fat deposition in the muscle. We studied 28 obese, sedentary postmenopausal Caucasian women. Women taking oral estrogen (n = 6) were matched for age (57 +/- 3 vs. 58 +/- 2 years), weight (87.9 +/- 6.0 vs. 83.0 +/- 3.9 kg), and BMI (33.9 +/- 1.7 vs. 33.9 +/- 1.9 kg/m(2)) with women not on HRT (n = 6). Eight women taking oral estrogen plus progesterone were matched with eight different women not on HRT for age (59 +/- 2 vs. 60 +/- 2 years), weight (82.8 +/- 3.7 vs. 83.7 +/- 4.1 kg), and BMI (30.7 +/- 1.0 vs. 29.9 +/- 1.3 kg/m(2)). VO(2max) (maximal aerobic capacity), percentage of fat, total body fat mass, and fat-free mass (FFM) were similar between groups. Visceral fat, subcutaneous abdominal fat, sagittal diameter, and mid-thigh low-density lean tissue (intramuscular fat) did not differ by hormone status. Basal carbohydrate and fat utilization was not different among groups. Fasting plasma glucose and insulin did not differ by hormone use. Glucose utilization (M) was measured during the last 30 min of a 3-h hyperinsulinemic-euglycemic clamp (40 mU. m(2). min(-1)). Postmenopausal women taking oral estrogen had a 31% lower M than women not on HRT (42.7 +/- 4.0 vs. 61.7 +/- 4.7 micromol. kg(FFM). min(-1), P < 0.05). M was 26% lower in women taking estrogen plus progesterone (44.0 +/- 3.5 vs. 59.7 +/- 6.2 micromol. kg(FFM). min(-1), P < 0.05) than women not on HRT. M/I, the amount of glucose metabolized per unit of plasma insulin (I), an index of insulin sensitivity, was 36% lower in women taking estrogen compared with matched women not on HRT (P < 0.05) and 28% lower in women taking estrogen plus progesterone compared with matched women not on HRT (P < 0.05). Postmenopausal women taking oral estrogen or those taking a combination of estrogen and HRT are more insulin-resistant than women not on HRT, even when women are of comparable total and abdominal adiposity.


Evaluation of radiolabeled (hetero)aromatic analogues of N-(2-diethylaminoethyl)-4-iodobenzamide for imaging and targeted radionuclide therapy of melanoma.
Targeted radionuclide therapy using radioiodinated compounds with a specific affinity for melanoma tissue is a promising treatment for disseminated melanoma, but the candidate with the ideal kinetic profile remains to be discovered. Targeted radionuclide therapy concentrates the effects on tumor cells, thereby increasing the efficacy and decreasing the morbidity of radiotherapy. In this context, analogues of N-(2-diethylaminoethyl)-4-iodobenzamide (BZA) are of interest. Various (hetero)aromatic analogues 5 of BZA were synthesized and radioiodinated with (125)I, and their biodistribution in melanoma-bearing mice was studied after i.v. administration. Most [ (125)I] 5-labeled compounds appeared to bind specifically and with moderate-to-high affinity to melanoma tumor. Two compounds, 5h and 5k, stood out with high specific and long-lasting uptake in the tumor, with a 7- and 16-fold higher value than BZA at 72 h, respectively, and kinetic profiles that makes them promising agents for internal targeted radionuclide therapy of melanoma.


Cassava (Manihot esculenta Crantz) and Yam (Dioscorea spp.) Crops and Their Derived Foodstuffs: Safety, Security and Nutritional Value.
Cassava (Manihot esculenta Crantz) and yam (Dioscorea spp.) are tropical crops consumed by ca. 2 billion people and represent the main source of carbohydrate and energy for the approximately 700 million people living in the tropical and sub-tropical areas. They are a guarantee of food security for developing countries. The production of these crops and the transformation into food-derived commodities is increasing, it represents a profitable business and farmers generate substantial income from their market. However, there are some important concerns related to the food safety and food security. The high post-harvest losses, mainly for yam, the contamination by endogenous toxic compounds, mainly for cassava, and the contamination by external agents (such as micotoxins, pesticides, and heavy metal) represent a depletion of economic value and income. The loss in the raw crops or the impossibility to market the derived foodstuffs, due to incompliance with food regulations, can seriously limit all yam tubers and the cassava roots processors, from farmers to household, from small-medium to large enterprises. One of the greatest challenges to overcome those concerns is the transformation of traditional or indigenous processing methods into modern industrial operations, from the crop storage to the adequate package of each derived foodstuff.


Dynamics of nematodes in a high organic loading rotating biological contactors.
Nematode diversity and dynamics of a full-scale rotating biological contactor plant (RBC) has been studied. Analysis of biofilm composition showed a well-established zoning of microfauna among the three RBC sections analysed. Nematodes appeared to be the dominant group within the larger microfauna populations with average abundances between 200 and 300ind/mg or 8000 and 17000ind/cm(2). The most abundant nematode species were Diplogasteritus nudicapitatus and Paroigolaimella coprophages and, to a lesser extent, Paroigolaimella bernensis and Steinernema intermedia. The relationship between nematodes and filamentous bacteria (specifically the genus Beggiatoa) was the most significant biotic relationship found, and to a lesser extent, nematodes with ciliates. The relationship between the abundance of nematode species and the physical-chemical variables suggests that nematodes may be good indicators of low pollutant load levels in the entry of the RBC system. Finally, the results indicate that nematodes may have a relevant role for a good biofilm development.


Inhibitory activity of cranberry extract on the bacterial adhesiveness in the urine of women: an ex-vivo study.
Strains of uropathogenic E. coli are responsible for approximately 90% of community-acquired, uncomplicated cystitis, and fimbriae represent the adhesive factors enabling E. coli to be anchored to uroepithelial cells in the first step of the infectious process. Recently, a few studies have shown that a correlation between the consumption of cranberry (Vaccinium macrocarpon) and prevention of UTI is related to the ability of proanthocyanidins to reduce the bacterial adhesion to uroepithelial cells. In this study we evaluate the inhibitory activity of urine of healthy women treated with tablets containing cranberry extract on the adhesiveness of E. coli to uroepithelial human cells. Two groups of 12 female volunteers each, aged between 18 and 65 years, were enrolled, one group with negative history and one group with positive history of recurrent cystitis. Subjects were treated with the active product or placebo in a random, cross-over, double-blinded sequence for one week in each of the two treatment sequences. Urine samples were collected at the beginning and the end of each study period. Tests of bacterial adhesiveness were performed with two strains of E. coli (ATCC 25922 and ATCC 35218) on HT1376 human bladder carcinoma cells. Significant reductions of bacterial adhesiveness were observed in women who received cranberry extract (-50.9%; p less than 0.0001), regardless of their medical history and the treatment period in the cross-over sequence. No changes were observed with placebo (-0.29%; n.s.). This ex-vivo study showed that the assumption of cranberry extract in suitable amounts can have an anti-adhesive activity on uropathogenic E. coli.


Limitations of nls beta-galactosidase as a marker for studying myogenic lineage or the efficacy of myoblast transfer.
Nuclear localizing beta-galactosidase (nls beta-gal) is used as a marker for studying myoblast cell lineage and for evaluating myoblast survival after myoblast transfer, a procedure with potential use for gene complementation for muscular dystrophy. Usefulness of this construct depends on the establishment of the extent to which nls beta-gal or its mRNA may be translocated from the nucleus that encodes it to other non-coding myonuclei in hybrid myofibers and the ease with which the encoding and non-coding myonuclei can be distinguished. Previous in vitro studies (Ralston and Hall 1989. Science, 244:1066-1068) have suggested limited translocation of the fusion protein. We re-examined the extent to which nls beta-gal is translocated in hybrid myofibers, both in vitro and in vivo, and evaluated the extent to which one can rely on histochemistry to distinguish encoding from non-coding nuclei in these myofibers. Myotubes formed in co-cultures of a myoblast line (MM14 cells), stably transfected with a construct consisting of a nls beta-gal under the control of the myosin light chain 3F promoter and 3' enhancer (3FlacZ10 cells), and [3H]-thymidine-labeled parental MM14 cells (plated at ratios of 1:6 or 1:20, respectively) were reacted with X-gal. After autoradiography, the distance over which nls beta-gal was translocated in hybrid myotubes was determined. In vivo translocation of nls beta-gal was evaluated by injecting [3H]-thymidine-labeled 3FlacZ10 myoblasts into the regenerating extensor digitorum longus muscle of immunosuppressed normal and mdx (dystrophin deficient) mice. Sections stained with X-gal and subjected to autoradiography permitted determination of the extent of nls beta-gal translocation in hybrid myofibers. In vitro: All nuclei in > 92% of hybrid myotubes showed evidence of nls beta-gal after exposure to X-gal, suggesting extensive translocation. Within hybrid myotubes, MM14-derived myonuclei approximately 350 microns from a 3FlacZ10-derived myonucleus showed evidence of nls beta-gal. In vivo: Similar translocation of nls beta-gal was observed in vivo. One week after myoblast transfer, donor-derived myonuclei were distinguishable from host-derived myonuclei containing nls beta-gal by the greater accumulation of reaction product in donor myonuclei after X-gal staining. However, 2 weeks after injection, host myonuclei often contained a significant amount of nls beta-gal, and accumulation of reaction product could not be used as the criterion for identification of donor myonuclei. Translocation of nls beta-gal (or its mRNA) is significantly greater than previously reported (Ralston and Hall 1989), resulting in large numbers of nls beta-gal positive non-coding myonuclei in hybrid myofibers. One week after myoblast transfer, distinguishing between nls beta-gal encoding and non-coding myonuclei in hybrid myofibers after X-gal staining of sectioned muscle is feasible; however, by 2 weeks, nls beta-gal increases in host myonuclei, making identification of donor-derived myonuclei problematic. Translocation of nls beta-gal to non-coding myonuclei in hybrid myofibers must be considered when nls beta-gal is used for studies of myogenic lineage or the efficacy of myoblast transfer therapy, particularly if long-term survival of hybrid myotubes is required.


Let-7d Inhibits Growth and Metastasis in Breast Cancer by Targeting Jab1/Cops5.
MicroRNAs (miRNAs) regulate the expressions of cancer-related genes, and are involved in the development and progression of various human cancers. Here, we performed further analyses to determine whether let-7d is functionally linked to Jab1 in breast cancer. In situ hybridization and immunohistochemical analyses were used to determine the level of let-7d and Jab1 in breast cancer clinical specimens and its correlation with clinicopathological data. Let-7d overexpressing breast cancer cell lines combined with mouse models bearing cell-derived xenografts were used to assess the functional role of let-7d both in vitro and in vivo. In this study, we found that let-7d was downregulated in breast cancer tissues, coupled with the elevations of Jab1 protein expressions, compared with paired adjacent noncancerous breast tissues. Let-7d overexpression significantly suppressed the proliferation and invasion in MCF-7 and MDA-MB-231 cells. Dual luciferase reporter assay indicated that Jab1 was the direct target of let-7d. Stepwise studies from in vitro and in vivo experiments indicated that let-7d overexpression inhibited cell growth and decreased Jab1 expressions in breast cancer cells and nude mice tumor tissues. Statistical analyses demonstrated that breast cancer patients with low levels of let-7d or high levels of Jab1 had a significant correlation with worse prognosis. These findings provide novel insights into molecular mechanism of let-7d and Jab1 in tumor development and progression of breast cancer, and thus let-7d/Jab1 are novel potential therapeutic targets for breast cancer patients.


[Fifty years after the enactment of the law to separate drug dispensing from doctors].
This report summarizes the action of Mr. Usho Mizoroki, who was not a pharmacist but was an activist for enacting the law to separate drug dispensing from doctors (Division of Labour Law). We collected the information Mrs. Shigeko Mizoroki revealed about her husband's activities towards enacting the Division of Labour Law, and added the historical background of that era. Nobody knows of his activities related to the enactment of the Division of Labour Law. It may be possible that the law itself was not successful, or that Mr. Usho Mizoroki was not a pharmacist. However it is important to know his efforts because the present division of labour practice is the result of the efforts of many people like him.


Clinically relevant interactions of anti-apoptotic Bcl-2 protein inhibitors with ABC transporters.
In this work we studied clinically relevant interactions between the BH3 mimetics and the ABCB1 and ABCG2 transporters. We observed that the intracellular levels of ABT-263 and ABT-199, but not ABT-737, might be reduced by ABCB1 or ABCG2. Importantly, this effect was proportional to the transporter expression level. High transporter expression levels decreased the intracellular levels of ABT-263 and ABT-199 substantially. Low transporter expression levels, which are clinically relevant, affected the intracellular level of ABT-263 slightly but significantly, however, they failed to decrease the intracellular level of ABT-199 below the control level in parental cells. Our results further revealed that ABT-263 did not inhibit the ABCB1 mediated transport, however, it partially inhibited the ABCG2 mediated transport at clinically relevant concentrations. In contrast, ABT-199 inhibited partially the ABCB1 mediated transport and it fully inhibited the ABCG2 mediated transport at clinically relevant concentrations. Importantly, cells expressing higher drug transporters levels required higher concentrations of ABT-263 or ABT-199 to achieve certain inhibition of substrate efflux. Antiproliferative effects of ABT-263 and ABT-199 might be reduced by ABCB1 or ABCG2, however, this effect depends on transporter expression levels. Since the expression levels of ABCB1 and ABCG2 are rarely high in clinical samples, their contribution to the overall resistance to ABT-263 or ABT-199 is probably low. Inhibition study revealed that ABT-199, but not ABT-263, fully inhibited low expression level of ABCG2. Our data suggest that ABT-199 should be evaluated beyond its original application as an inhibitor of the ABCG2 transporter in clinical settings.


Determination and bioimaging method for nitric oxide in biological specimens by diaminofluorescein fluorometry.
A simple and sensitive assay and a cellular bioimaging method for nitric oxide (NO) were developed using a novel diaminofluorescein DAF-FM and its diacetate. DAF-FM is converted via an NO-specific mechanism to an intensely fluorescent triazole derivative. For the measurement of NO, the triazole derivative of DAF-FM was determined by reversed-phase high-performance liquid chromatography with fluorescence detection. In the presence of 1 microM DAF-FM, the concentrations of NOR-1, an NO donor, in the range of 2-200 nM were linearly related to the fluorescence intensity. This sensitive NO assay enabled us to detect the spontaneous and substance P-induced NO release from isolated porcine coronary arteries, both of which were dependent entirely on the NO synthase activity in vascular endothelial cells. We also obtained fluorescence images of cultured smooth muscle cells of the rat urinary bladder after loading with DAF-FM diacetate. In the cells pretreated with cytokines, the fluorescence intensity increased with time after DAF-FM loading. This increase in the fluorescence intensity was blocked by prior treatment of the muscle cells with an NO synthase inhibitor, N(G)-nitro-l-arginine methyl ester. Therefore, the present novel diaminofluorescein fluorometry should be useful not only for sensitive NO assay, but also for NO imaging in a variety of biological specimens.


Artificial nutritional support in cancer patients after esophagectomy: 11 years of experience.
Esophageal cancer represents a high-risk group of patients. This study determines the association of artificial nutrition with morbidity, mortality, and survival and studies clinical situations that determine the choice between enteral (EN) and parenteral support (PN). This retrospective single-center study compared 2 periods: 1) treatment centered in surgical process with discretionary demand of support, and 2) elective therapeutic and nutritional interventions were systematized. Risks factors that determined use of PN and survival were included in 4 multivariate regression models: 2 logistic, 1 multinomial, and a survival Cox analysis. Significance determined with 95% confidence interval (CI) of 95%; inclusion criteria was P < 0.1. During an 11-yr period, 175 patients were studied. Artificial nutrition consisted of 45 jejunostomy EN, 28 PN, and 102 both. Risk factors that conditioned PN were first period (OR: 2.41; 95% CI: 1.13-5.14), stay in intensive care unit (ICU) >3 days (OR: 1.70; 95% CI: 0.93-3.71), and surgical reintervention (OR: 3.83; 95% CI: 0.94-16.95). Risk factors associated with mortality were first period (OR: 22.7; 95% CI: 2.31-172.05), respiratory infection (OR: 11.23; 95% CI: 2.33-55.5) and coloplasty surgery (OR: 13.16; 95% CI: 2.11-83.33). Longer survival was associated with second period (OR: 2.36; 95% CI: 1.38-4.05) and lower neoplasm staging (OR: 1.43; 95% CI: 1.21-1.69). A multidisciplinary management that includes nutritional support of esophagectomized patients is 1 of the factors that improves survival. Protocol implies greater use of EN; PN remains an important nutritional therapy.


Refinement of the spinal muscular atrophy locus to the interval between D5S435 and MAP1B.
The childhood-onset SMA locus has been mapped to chromosome 5q13, in a region bounded by the proximal locus, D5S6, and the closely linked distal loci, D5S112 and MAP1B. We now describe a highly polymorphic, tightly linked microsatellite marker (D5S435) that is very likely the closest proximal marker to the SMA locus. Multipoint linkage analysis firmly establishes the following order of markers at 5q13: centromere-D5S76-D5S6-D5S435-MAP1B/D5S112- D5S39-telomere. The data indicate that SMA resides in an approximately 0.7-cM (range 0.1-2.1) region between D5S435 and MAP1B. This finding reduces by approximately fourfold the genetic region that most likely harbors the SMA locus and will facilitate the physical mapping and cloning of the disease gene region.


[Neuroendocrine sarcoidosis: a case].
The authors report a case of neuro-endocrine sarcoidosis, associating panhypopituitarism , diabetes insipidus, and involvement of the optic chiasma with behavioural changes, chronic meningitis and a diffuse pyramidal syndrome. The diagnosis was suggested by the presence of mediastinal lymphadenopathy and confirmed during a surgical procedure to conserve the optic nerves. The clinical course on corticosteroids was unfavourable because of iatrogenic cushing' syndrome and steroid dependence . It was not possible to withdraw steroids, even with the use of immunosuppressors (Methotrexate).


Transsphenoidal resection of pituitary adenoma: using a critical pathway.
The use of a transsphenoidal critical pathway can be a vital tool for critical care nurses in anticipating complications and improving patient outcomes. Complications such as diabetes insipidus and cerebrospinal fluid leak associated with posttranssphenoidal patients may result in prolonged hospitalization and worsened functional outcome. Implementing a transsphenoidal critical pathway for these patients can guide their care and alert critical care nurses to potential complications and their prevention and/or treatment.


Characterization of gyrA and gyrB mutations associated with fluoroquinolone resistance in Mycobacterium tuberculosis isolates from Morocco.
Fluoroquinolones (FQs) are the cornerstone of treatment for drug-resistant tuberculosis (TB). They are the most effective second-line antimycobacterial drugs and are recommended for the treatment of multidrug-resistant TB (MDR-TB). However, it is widely accepted that FQ resistance is high among MDR-TB isolates. Thus, characterisation of mutations conferring resistance to FQs will be of a great interest for effective and efficient management of TB resistance in Morocco. A laboratory collection of 30 Mycobacterium tuberculosis isolates previously characterised as phenotypically and genotypically MDR as well as 20 randomly selected pan-susceptible isolates were included in this retrospective study. The mutation profiles associated with resistance to FQs were assessed by PCR and DNA sequencing. Target sequences for two genes (gyrA and gyrB) were examined. All strains had their fingerprint previously established by spoligotyping. Molecular analyses showed that 30% of the MDR-TB isolates harboured FQ resistance mutations in gyrA, with the most prevalent being an alanine to threonine at position 90 (Ala90Thr) (56%; 5/9). None of the isolates harboured mutations in gyrB. All gyrA resistance mutant strains belonged to the LAM lineage, mostly LAM9, raising the possible emergence of a specific clone (gyrA mutant/LAM9). The results of this preliminary study highlight the high prevalence of FQ resistance among MDR-TB isolates in Morocco and consequently the need for rapid detection of FQ resistance once MDR-TB is confirmed to adjust treatment in a timely manner and to interrupt the propagation of more severe forms of M. tuberculosis drug resistance.


Cryopreservation of lar gibbon semen collected by manual stimulation.
We confirmed ejaculation as a result of manual stimulation in a lar gibbon, and attempted to cryopreserve the semen using TES-Tris-egg yolk-based (TTE) extender. After measuring the amount of semen (g), we first diluted the semen with TTE extender, and calculated sperm concentration (sperm/ml), total sperm count (sperm), and progressive sperm motility (%). Then, we cooled diluted semen slowly to 4 °C over 2 h, and added an equal volume of secondary extender containing glycerol over 30 min. Finally, we flash-froze the semen solution by plunging into liquid nitrogen. In addition, we freeze-thawed the solution to determine the recovery rate of the motile sperm. Collection of semen was successful on four of the five occasions. The median (min-max) quantity of ejaculate was 0.19 g (0.09-0.26 g), the median sperm concentration was 1.38 × 10(9) sperm/ml (1.20-1.53 × 10(9) sperm/ml), and the median total sperm count was 0.26 × 10(9) sperm (0.11-0.40 × 10(9) sperm). Moreover, the median sperm motility immediately after ejaculation was 65 % (60-75 %), the median sperm motility after freeze-thawing was 30 % (25-35 %), and the median recovery rate was 42.3 % (40.0-58.3 %). We were able to (1) collect semen from a lar gibbon by manual stimulation, (2) reveal andrological findings regarding semen characteristics, and (3) preserve the genetic resource using TTE cryopreservation.