Sea Spray Aerosol: The Chemical Link between the Oceans, Atmosphere, and Climate.
The oceans, atmosphere, and clouds are all interconnected through the release and deposition of chemical species, which provide critical feedback in controlling the composition of our atmosphere and climate. To better understand the couplings between the ocean and atmosphere, it is critical to improve our understanding of the processes that control sea spray aerosol (SSA) composition and which ones plays the dominate role in regulating atmospheric chemistry and climate.


[Nosocomial urinary tract infection: an analysis beyond urinary catheterization].
Nosocomially acquired urinary tract infections (NAUTI) represent an important public health issue, but its characteristics when they are not catheter associated (CA-UTI) or when they take place outside intensive care units (ICU) are poorly understood. To determine the patients' characteristics, etiology and antimicrobial susceptibility of NAUTI, both CA-UTI and no CA-UTI, in general ward and ICU. We conducted a retrospective analytic cross-sectional study, between 2009 and 2013, in a third level universitary hospital. All NAUTI episodes were identified, classifying them as CA-UTI and no CA-UTI. We included 253 episodes of NAUTI, being CA-UTI (60,9%) more frequent than no CA-UTI. A 37,4% of no CA-UTI and 59,7% of CA-UTI were identified in ICU. The most frequently isolated microorganisms were Escherichia coli, Klebsiella pneumoniae and Enterococcus sp. A 19% of extended spectrum betalactamase producing gram negative bacilli were found, without differences between groups. Patients's comorbidities, microorganisms associated to NAUTI and its antimicrobial susceptibility were similar in CA-UTI and no CA-UTI, as in general ward and ICU.


Measurement of calcium influx in tethered rings of rabbit aorta under tension.
Calcium (Ca) influx in vascular smooth muscle is routinely measured in untethered preparations not under passive stretch, and Ca influx data are correlated with data for steady-state isometric tension obtained under parallel conditions from tethered preparations under passive stretch. The validity of this method was tested by simultaneous measurement of Ca influx and tension in tethered rings of rabbit thoracic aorta. Ca influx (45Ca 3-min pulse) and tension were measured at 3 and 30 min after norepinephrine (NE) or KCl and under control (no agonist) conditions. Active tension was significantly altered by variations in passive tension. Ca influx was unaffected by passive tension under control, NE, or KCl conditions, and results were similar at 3 and 30 min. The results confirm the validity of correlating Ca influx data from untethered rings with steady-state contractile response data obtained from tethered rings under similar experimental conditions.


Social capital and social exclusion in England and Wales (1972-1999).
Recent research on social capital has explored trends in membership in voluntary organizations. However, there is currently little robust evidence on such trends in the UK since the 1970s, nor is there any analysis of whether participation bridges social divisions or accentuates them. This paper explores trends in participation in England and Wales since 1972 using data from the Social Mobility Inquiry of 1972 and the British Household Panel Survey of 1992 and 1999. We are concerned with social exclusion mechanisms in social capital generation in Britain over the three decades. Using binomial and multinomial models to 'unpack' the effects of socio-cultural factors on civic participation and on different types of associational membership, we test the thesis of across-the-board decline in social capital by Putnam (2000) and that of rising levels of middle-class social capital versus consistent low levels of working-class social capital by Hall (1999). The results show significant socio-cultural-gender differences, a relative stability of middle-class participation, and a rapid decline in the working-class access to social capital. We challenge the established accounts of both theses.


Lethal and Sublethal Effects of Three Microbial Biocontrol Agents on Spodoptera litura and Its Natural Predator Rhynocoris kumarii.
Entomopathogenic microbes such as Spodoptera litura nucleopolyhedrovirus (SpltNPV), Metarhizium anisopliae, and Pseudomonas fluorescens are biological agents used for the control of multiple arthropod pests. The objective of this study was to assess their effects on the biological parameters of Spodoptera litura (Lepidoptera: Noctuidae) larvae, and its natural reduviid predator Rhynocoris kumarii (Hemiptera: Reduviidae) under laboratory conditions. Results suggested that P. fluorescens reduced the food consumption index, relative growth rate, approximate digestibility, the efficiency of conversion of ingested food, and the efficiency of conversion of digested food of S. litura third instar larvae compared to prey infected with M. anisopliae and SpltNPV. Both SpltNPV and M. anisopliae caused similar mortality of S. litura life stages after 96 h of observation. To observe the effect of an infected prey diet on predator behavior, infected S. litura larvae were offered to the third, fourth, and fifth instar nymphs of R. kumarii, and their prey handling time, predation rate (number/day/predator), developmental period, and the survival rate was recorded. When the life stages of R. kumarii were offered entomopathogen-infected S. litura larvae, their predation rate was comparable to or higher than the untreated control. The juvenile predator, after feeding on P. fluorescens-infected S. litura larvae, had a significantly longer developmental period (2⁻4 days) compared to those fed on larvae infected with other microbial control agents. However, feeding on P. fluorescens alone did not affect the predator nymphal survival rate or the adult sex ratio. Although three entomopathogens had some degree of effect on the biological parameters of R. kumarii, the outcome of this study suggests that integration of reduviids with the tested entomopathogens are a compatible and potentially effective strategy for the management of S. litura populations. However promising, this combined strategy needs to be tested under field conditions to confirm the laboratory findings.


[Cell cycle and cancer--from the tumor cell line to the patient].
In normal cells the cell cycle is precisely regulated by a set of different gene products. Cyclins move cells along into the cycle while cyclin-dependent kinase inhibitors function as cell cycle brakes. Mutations in such genes are frequent in cancer and may exert a profound effect on their biology. Overactive cyclins may push cells into inadequate cycling. Inactivation of cell cycle brakes might likewise provide a proliferation advantage to tumour cells over normal cells. Basic research in cell biology and molecular biology has greatly enlarged our knowledge in this timely field. The detection of gene alterations impinging on the cell cycle machinery in cancer is of increasing practical relevance. Mutated genes such as the inactivated p53 tumour suppressor gene may be prognostically relevant in human cancer, or predict the appropriate choice of treatment. Precise knowledge of cell cycle disturbances will eventually pave the way towards gene therapy of cancer although achievement of this goal is still in the distant future.


The impact of maternal childhood abuse on parenting and infant temperament.
The impact of maternal history of maltreatment and psychopathology on mother-child relationship, parenting, and infant temperament was evaluated. Women completed self-report measures addressing childhood trauma, psychopathology, infant-parent relationship, parenting, and infant temperament. Maternal physical abuse was associated with poorer mother-child interactions, increased vigilance, and difficulty recovering from distress among infants, whereas a history of emotional abuse was linked with less interactional dysfunction, lower levels of infant frustration, and more pleasure. Maternal depression was associated with infant temperament and attitudes about parenting. These findings suggest that maternal history of childhood abuse and psychopathology are important determinants of parenting and infant temperament.


Development of an esophagus acellular matrix tissue scaffold.
A cell-extraction protocol yielding an esophagus acellular matrix (EAM) scaffold for use in tissue engineering of an esophagus, including hypotonic lysis, multiple detergent cell extraction steps, and nucleic acid digestion, was developed in a rat model. Histological techniques, burst pressure studies, in vitro esophageal epithelial cell seeding, and in vivo implantation were used to assess cell extraction, extracellular matrix (ECM) preservation, and biocompatibility. Microscopy demonstrated that cell extraction protocols using sodium dodecyl sulfate (SDS) (0.5%, wt/vol) as a detergent resulted in cell-free EAM with retained ECM protein collagen, elastin, laminin, and fibronectin. Burst pressure studies indicated a loss of tensile strength in EAMs, but at intraluminal pressures that were unlikely to affect in vivo application. In vitro cell seeding studies exhibited epithelial cell proliferation with stratification similar to native esophagi after 11 days, and subcutaneously implanted EAMs displayed neovascularization and a minimal inflammatory response after 30 days of implantation. This study presents an esophagus acellular matrix tissue scaffold with preserved ECM proteins, biomechanical properties, and the ability to support esophageal cell proliferation to serve as the foundation for a tissue-engineered esophagus.


Overexpression of GCN2-type protein kinase in wheat has profound effects on free amino acid concentration and gene expression.
A key point of regulation of protein synthesis and amino acid homoeostasis in eukaryotes is the phosphorylation of the α subunit of eukaryotic translation initiation factor 2 (eIF2α) by protein kinase general control nonderepressible (GCN)-2. In this study, a GCN2-type PCR product (TaGCN2) was amplified from wheat (Triticum aestivum) RNA, while a wheat eIF2α homologue was identified in wheat genome data and found to contain a conserved target site for phosphorylation by GCN2. TaGCN2 overexpression in transgenic wheat resulted in significant decreases in total free amino acid concentration in the grain, with free asparagine concentration in particular being much lower than in controls. There were significant increases in the expression of eIF2α and protein phosphatase PP2A, as well as a nitrate reductase gene and genes encoding phosphoserine phosphatase and dihydrodipicolinate synthase, while the expression of an asparagine synthetase (AS1) gene and genes encoding cystathionine gamma-synthase and sulphur-deficiency-induced-1 all decreased significantly. Sulphur deficiency-induced activation of these genes occurred in wild-type plants but not in TaGCN2 overexpressing lines. Under sulphur deprivation, the expression of genes encoding aspartate kinase/homoserine dehydrogenase and 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase was also lower than in controls. The study demonstrates that TaGCN2 plays an important role in the regulation of genes encoding enzymes of amino acid biosynthesis in wheat and is the first to implicate GCN2-type protein kinases so clearly in sulphur signalling in any organism. It shows that manipulation of TaGCN2 gene expression could be used to reduce free asparagine accumulation in wheat grain and the risk of acrylamide formation in wheat products.


Adult height in girls with central precocious puberty without gonadotropin-releasing hormone agonist treatment: a retrospective case-control study.
The primary aim of this study was to investigate the final adult height (FAH) of girls diagnosed with central precocious puberty (CPP) who were untreated. We retrospectively analyzed the medical records of 36 girls diagnosed with CPP between 8 and 9 years of age who did not receive treatment, and 206 girls diagnosed with CPP within the same age range who received gonadotropin-releasing hormone (GnRH) agonist treatment. Midparental height (MPH), predicted adult height (PAH) obtained using height and bone age (BA) at the time of diagnosis (PAH for BA), and PAH obtained using the Bayley-Pinneau method (PAH by BP) were calculated. Additionally, height at the time of growth completion was compared with the predicted height. The FAHs were 160.71±4.56 cm in the untreated group and 159.31±4.26 cm in the treated group. In the untreated group, the FAH was 0.99±4.50 cm shorter than the MPH but 4.29±3.33 cm and 3.46±3.93 cm greater than the PAH for BA and PAH by BP, respectively. In children diagnosed with CPP between 8 and 9 years of age who were untreated, FAH was greater than PAH for BA and PAH by BP at the time of diagnosis, indicating that the prognosis of FAH was not poor. Therefore, for girls diagnosed with CPP, it is recommended to consider various conditions, such as pubertal onset, height at diagnosis, BA, peak luteinizing hormone level, predicted height, and speed of puberty, when deciding whether to administer GnRH agonists.


Heat shock of human synovial and dermal fibroblasts induces delayed up-regulation of collagenase-gene expression.
We investigated the effect of heat shock on the expression of the collagenase gene in normal human synovial and dermal fibroblasts. Heat shock (42-44 degrees C for 1 h) caused a marked increase in heat-shock protein 70 (HSP-70) mRNA levels, followed by a delayed increase in collagenase mRNA levels, in both cell types. Pretreatment with cycloheximide had no effect on the heat-shock-induced increase in HSP-70 mRNA expression, but abrogated the induction of collagenase mRNA during the recovery. To study the mechanisms of collagenase-gene induction by heat shock, the transcriptional activity of a collagenase-promoter-driven chloramphenicol acetyltransferase (CAT) reporter gene was examined in transient transfection experiments. Heat shock was followed by a > 2-fold increase in CAT activity driven by a 3.8 kb fragment of the collagenase promoter, or by a construct containing an AP-1 binding site. A mutation in the AP-1 binding site abolished the effect of heat shock. Electrophoretic-mobility-shift assays revealed a marked increase in DNA-binding activity specific for the AP-1 binding site in nuclear extracts prepared from synovial fibroblasts recovering from heat shock. These results indicate that heat shock causes a delayed increase in collagenase-gene expression in human fibroblasts, and suggests that this stimulation involves, at least in part, transcriptional activation through an AP-1 binding site. Heat shock appears to initiate a programme of cellular events resulting in collagenase-gene expression, and therefore may contribute to connective-tissue degradation in disease states.


Model-Based Design and Optimization of Blood Oxygenators.
Blood oxygenators, also known as artificial lungs, are widely used in cardiopulmonary bypass surgery to maintain physiologic oxygen (O2) and carbon dioxide (CO2) levels in blood, and also serve as respiratory assist devices to support patients with lung failure. The time- and cost-consuming method of trial and error is initially used to optimize the oxygenator design, and this method is followed by the introduction of the computational fluid dynamics (CFD) that is employed to reduce the number of prototypes that must be built as the design is optimized. The CFD modeling method, while having progress in recent years, still requires complex three-dimensional (3D) modeling and experimental data to identify the model parameters and validate the model. In this study, we sought to develop an easily implemented mathematical models to predict and optimize the performance (oxygen partial pressure/saturation, oxygen/carbon dioxide transfer rates, and pressure loss) of hollow fiber membrane-based oxygenators and this model can be then used in conjunction with CFD to reduce the number of 3D CFD iteration for further oxygenator design and optimization. The model parameters are first identified by fitting the model predictions to the experimental data obtained from a mock flow loop experimental test on a mini fiber bundle. The models are then validated through comparing the theoretical results with the experimental data of seven full-size oxygenators. The comparative analysis show that the model predictions and experimental results are in good agreement. Based on the verified models, the design curves showing the effects of parameters on the performance of oxygenators and the guidelines detailing the optimization process are established to determine the optimal design parameters (fiber bundle dimensions and its porosity) under specific system design requirements (blood pressure drop, oxygen pressure/saturation, oxygen/carbon dioxide transfer rates, and priming volume). The results show that the model-based optimization method is promising to derive the optimal parameters in an efficient way and to serve as an intermediate modeling approach prior to complex CFD modeling.


Colocalization in vivo and association in vitro of perlecan and elastin.
We have colocalized elastin and fibrillin-1 with perlecan in extracellular matrix of tensional and weight-bearing connective tissues. Elastin and fibrillin-1 were identified as prominent components of paraspinal blood vessels, and posterior longitudinal ligament in the human fetal spine and outer annulus fibrosus of the fetal intervertebral disc. We also colocalized perlecan with a synovial elastic basal lamina, where the attached synovial cells were observed to produce perlecan. Elastin, fibrillin-1 and perlecan were co-localized in the intima and media of small blood vessels in the synovium and in human fetal paraspinal blood vessels. Elastic fibers were observed at the insertion point of the anterior cruciate ligament to bone in the ovine stifle joint where they colocalized with perlecan. Elastin has not previously been reported to be spatially associated with perlecan in these tissues. Interactions between the tropoelastin and perlecan heparan sulfate chains were demonstrated using quartz crystal microbalance with dissipation solid phase binding studies. Electrostatic interactions through the heparan sulfate chains of perlecan and core protein mediated the interactions with tropoelastin, and were both important in the coacervation of tropoelastin and deposition of elastin onto perlecan immobilized on the chip surface. This may help us to understand the interactions which are expected to occur in vivo between the tropoelastin and perlecan to facilitate the deposition of elastin and formation of elastic microfibrils in situ and would be consistent with the observed distributions of these components in a number of connective tissues.


Tissue distribution and biochemical and functional properties of Tp55 (CD27), a novel T cell differentiation antigen.
Two monoclonal antibodies (CLB-CD 27/1 and CLB-CD 27/2) were raised against a novel determinant on human T lymphocytes. One of these antibodies, CLB-CD 27/1 (clone 9F4), was grouped by the Third International Workshop and Conference on Human Leucocyte Differentiation Antigens together with three other monoclonal antibodies (VIT 14, OKT 18A, and S152) in the new cluster CD27. In this paper we show that antibodies belonging to this cluster recognize an antigen present on a large subset of peripheral T lymphocytes and most medullary thymocytes. At least two different nonoverlapping epitopes were identified with directly labeled monoclonal antibodies. Immunoprecipitation studies indicate that the target antigen of CD27 antibodies is a polypeptide of 55 kDa, which appears in the form of a disulfide-linked homodimer on the T lymphocyte membrane (Tp55). Stimulation of T cells via the T3/T cell antigen-receptor complex, with either phytohemagglutinin or CD3 monoclonal antibodies, resulted in a fivefold increase in the membrane expression of Tp55, whereas activation by phorbol myristate acetate caused a marked down-regulation. Moreover, an additional molecule of 32 kDa was precipitated from the membrane of activated but not of resting T cells. Addition of CD27 antibodies to cultures stimulated with either phytohemagglutinin or CD3 monoclonal antibody led to enhanced proliferation, whereas no effect was observed in phorbol myristate acetate or interleukin 2-stimulated cultures. The possible role of the Tp55 antigen in T cell activation is discussed.


Differences in community and academic practice patterns for newly diagnosed myelodysplastic syndromes (MDS) patients.
The heterogeneous nature of myelodysplastic syndromes (MDS) complicates therapeutic decision making, particularly for newly diagnosed disease. Factors impacting the treatment plan in this early period of disease course are poorly defined. This study determines whether therapeutic choices for newly diagnosed MDS are associated with location of treatment (community or academic), prognostic risk category, and patient age. The adults in Minnesota with myelodysplastic syndromes (AIMMS) database was utilized in this statewide, prospective population-based study conducted by the University of Minnesota (UMN), Mayo Clinic, and Minnesota Department of Health. Adult (age 20+ years) cases of MDS newly diagnosed starting in April 2010 were invited to participate. This analysis includes patients enrolled during the first study year with 1-year follow-up data. Treatment choices (supportive, active, and transplant) were stratified by the international prognostic scoring system (IPSS) and the revised-IPSS (IPSS-R), then separated into groups by location of care and age (<65 or 65+ years). Academic-based care was any contact with the UMN and Mayo Clinic; community-based care was all other clinical sites. Stratification by IPSS and IPSS-R showed supportive care decreased and active care increased with advancing risk categories (p<0.0001). Comparing treatment setting, community-based care had 77% supportive and 23% active treatment; academic-based care was 36% supportive, 41% active, and 23% transplant (p<0.0001). By age groups, patients <65 years with intermediate, high, or very high risk disease by IPSS-R received 97% active care/transplant, compared to only 52% of patients age 65+. Younger patients and those treated at academic centers had a more aggressive treatment approach. Whether these treatment differences convey improved disease control and mortality, and therefore should be extended more frequently to older and community-based patients, is the subject of ongoing prospective study.


Epilepsy surgery in India.
Modern epilepsy started in India in 1995 at Sri Chitra Tirunal Institute of Medical Science and Technology, Trivandrum and at All India Institute of Medical Sciences, New Delhi. At both centres the attempt was to get the program going with patients having surgically remediable epilepsy syndromes -who could be evaluated with non invasive investigations. The mainstay of the evaluation was a good quality epilepsy specific MRI and video EEG coupled with a SPECT study and a neuropsychological evaluation. Concordance of the focus on all investigations was critical to a good outcome. There were several problems on the way - but they were managed keeping in consideration our local needs and requirements. Intraoperative electocorticography was done and good outcomes attained. The critical determinants of success were the formation of a team with various interdisciplinary specialists and a strong will to succeed.


[A man in his eighties with recurrent syncope].
Syncope is usually a benign event that affects up to 50% of people over a lifetime, needing no extensive examination. The challenge is to diagnose the few with underlying life-threatening disease in need of immediate medical attention. Guidelines are clear, but unfortunately, clinical practice does not always follow recommendations, as illustrated by this case report. A diabetic, hypertensive male in his eighties had a myocardial infarction in his medical history. He presented to his GP with recurrent syncopal episodes that had occurred while erect and in motion. Physical examination was found to be normal for his age, and he was referred to a cardiologist and seen three months later. An ECG showed sinus rhythm and a previous inferior wall myocardial infarction. A 24-hour ECG recorder was fitted, and a consultation was scheduled for the next day. At home the patient died. The ECG recording revealed sinus rhythm with increasing depression of the ST segment, followed by rapidly conducted atrial fibrillation, and then rapid ventricular tachycardia followed by terminal ventricular fibrillation. A patient with a history where there is suspicion of cardiac syncope should be immediately and intensively examined when presenting to the healthcare services.


Clinics in diagnostic imaging (6).
A nine-month-old Chinese girl presented with a large buttock mass. Serum alpha-fetoprotein level was elevated. Plain radiographs, computerised tomography and magnetic resonance imaging demonstrated a solid heterogeneous presacral mass. A sacrococcygeal teratoma was excised using the abdominoperineal approach. The patient responded well to multiagent chemotherapy. The features of this tumour are discussed, with emphasis on the role of imaging.


Identifying patients at risk for nonroutine discharge after surgery for cervical myelopathy: an analysis from the Quality Outcomes Database.
Optimizing patient discharge after surgery has been shown to impact patient recovery and hospital/physician workflow and to reduce healthcare costs. In the current study, the authors sought to identify risk factors for nonroutine discharge after surgery for cervical myelopathy by using a national spine registry. The Quality Outcomes Database cervical module was queried for patients who had undergone surgery for cervical myelopathy between 2016 and 2018. Nonroutine discharge was defined as discharge to postacute care (rehabilitation), nonacute care, or another acute care hospital. A multivariable logistic regression predictive model was created using an array of demographic, clinical, operative, and patient-reported outcome characteristics. Of the 1114 patients identified, 11.2% (n = 125) had a nonroutine discharge. On univariate analysis, patients with a nonroutine discharge were more likely to be older (age ≥ 65 years, 70.4% vs 35.8%, p < 0.001), African American (24.8% vs 13.9%, p = 0.007), and on Medicare (75.2% vs 35.1%, p < 0.001). Among the patients younger than 65 years of age, those who had a nonroutine discharge were more likely to be unemployed (70.3% vs 36.9%, p < 0.001). Overall, patients with a nonroutine discharge were more likely to present with a motor deficit (73.6% vs 58.7%, p = 0.001) and more likely to have nonindependent ambulation (50.4% vs 14.0%, p < 0.001) at presentation. On multivariable logistic regression, factors associated with higher odds of a nonroutine discharge included African American race (vs White, OR 2.76, 95% CI 1.38-5.51, p = 0.004), Medicare coverage (vs private insurance, OR 2.14, 95% CI 1.00-4.65, p = 0.04), nonindependent ambulation at presentation (OR 2.17, 95% CI 1.17-4.02, p = 0.01), baseline modified Japanese Orthopaedic Association severe myelopathy score (0-11 vs moderate 12-14, OR 2, 95% CI 1.07-3.73, p = 0.01), and posterior surgical approach (OR 11.6, 95% CI 2.12-48, p = 0.004). Factors associated with lower odds of a nonroutine discharge included fewer operated levels (1 vs 2-3 levels, OR 0.3, 95% CI 0.1-0.96, p = 0.009) and a higher quality of life at baseline (EQ-5D score, OR 0.43, 95% CI 0.25-0.73, p = 0.001). On predictor importance analysis, baseline quality of life (EQ-5D score) was identified as the most important predictor (Wald χ2 = 9.8, p = 0.001) of a nonroutine discharge; however, after grouping variables into distinct categories, socioeconomic and demographic characteristics (age, race, gender, insurance status, employment status) were identified as the most significant drivers of nonroutine discharge (28.4% of total predictor importance). The study results indicate that socioeconomic and demographic characteristics including age, race, gender, insurance, and employment may be the most significant drivers of a nonroutine discharge after surgery for cervical myelopathy.


Adaptive geometry image.
We present a novel post-processing utility called adaptive geometry image (AGIM) for global parameterization techniques that can embed a 3D surface onto a rectangular1 domain. This utility first converts a single rectangular parameterization into many different tessellations of square geometry images(GIMs) and then efficiently packs these GIMs into an image called AGIM. Therefore, undersampled regions of the input parameterization can be up-sampled accordingly until the local reconstruction error bound is met. The connectivity of AGIM can be quickly computed and dynamically changed at rendering time. AGIM does not have T-vertices, and therefore no crack is generated between two neighboring GIMs at different tessellations. Experimental results show that AGIM can achieve significant PSNR gain over the input parameterization, AGIM retains the advantages of the original GIM and reduces the reconstruction error present in the original GIM technique. The AGIM is also for global parameterization techniques based on quadrilateral complexes. Using the approximate sampling rates, the PolyCube-based quadrilateral complexes with AGIM can outperform state-of-the-art multichart GIM technique in terms of PSNR.


Vigabatrin.
Vigabatrin (VGB) is a structural analogue of the inhibitory neurotransmitter gamma-amino butyric acid (GABA), which produces its antiepileptic effect by irreversibly inhibiting the degradative enzyme GABA-transaminase. This produces an increase in central nervous system (CNS) GABA levels. VGB is among the few antiepileptic drugs (AEDs) that was synthesized with a specific targeted mechanism in mind and was subsequently demonstrated to function by that mechanism. Tiagabine, a GABA reuptake blocker, is the only other "designer drug" among the currently available AEDs. Therefore, VGB is among the few AEDs for which the mechanism of action is well understood. Recently, safety issues have been raised with regard to the use of vigabatrin. This article reviews the mechanism of action, pharmacokinetics, safety, and efficacy of VGB.


Therapeutic management of cystic fibrosis.
The therapeutic management of patients with cystic fibrosis (CF) is reviewed. CF is an autosomal-recessive genetic disease that affects all exocrine glands. Drug therapy is directed toward the relief of symptoms and prevention of complications. Chronic respiratory tract infections and pancreatic insufficiency are the major manifestations. The replacement of pancreatic enzymes is best achieved with microencapsulated preparations. The dose of enzymes is increased until a maximum effect on decreasing steatorrhea is observed. Pseudomonas aeruginosa and Staphylococcus aureus are the most common pathogens in respiratory infections, which usually require 2-3 weeks of intravenous antibiotic therapy. Because patients with CF often have rapid clearance of both penicillins and aminoglycosides and altered apparent distribution volumes of aminoglycosides, individualized dosing is required. The role of prophylactic antibiotic therapy is controversial, since prolonged exposure to antimicrobials may hasten the development of mucoid strains. Aerosolized antibiotics have not been consistently useful in treating these patients, and there is no convincing evidence to support their long-term use. There are also no clear data to support the use of oral or aerosolized mucolytic agents. The inability to deliver the drug to the site of action plus the time and expense of inhalation equipment may only complicate the drug therapy while producing little benefit. Bronchodilators can be useful in improving pulmonary function. Vitamin supplementation with both the fat- and water-soluble vitamins is important in the treatment of CF. Iron supplementation may also be necessary. Until the basic defect in CF is discovered, the use of antibiotics and replacement of pancreatic enzymes will continue to be the mainstay of therapy.


Direct synthesis of Fe3 C-functionalized graphene by high temperature autoclave pyrolysis for oxygen reduction.
We present a novel approach to direct fabrication of few-layer graphene sheets with encapsulated Fe3 C nanoparticles from pyrolysis of volatile non-graphitic precursors without any substrate. This one-step autoclave approach is facile and potentially scalable for production. Tested as an electrocatalyst, the graphene-based composite exhibited excellent catalytic activity towards the oxygen reduction reaction in alkaline solution with an onset potential of ca. 1.05 V (vs. the reversible hydrogen electrode) and a half-wave potential of 0.83 V, which is comparable to the commercial Pt/C catalyst.


Transmembrane-domain determinants for SNARE-mediated membrane fusion.
Neurosecretion involves fusion of vesicles with the plasma membrane. Such membrane fusion is mediated by the SNARE complex, which is composed of the vesicle-associated protein synaptobrevin (VAMP2), and the plasma membrane proteins syntaxin-1A and SNAP-25. Although clearly important at the point of membrane fusion, the precise structural and functional requirements for the transmembrane domains (TMDs) of SNAREs in bringing about neurosecretion remain largely unknown. Here, we used a bimolecular fluorescence complementation (BiFC) approach to study SNARE protein interactions involving TMDs in vivo. VAMP2 molecules were found to dimerise through their TMDs in intact cells. Dimerisation was abolished when replacing a glycine residue in the centre of the TMD with residues of increasing molecular volume. However, such mutations still were fully competent in bringing about membrane-fusion events, suggesting that dimerisation of the VAMP2 TMDs does not have an important functional role. By contrast, a series of deletion or insertion mutants in the C-terminal half of the TMD were largely deficient in supporting neurosecretion, whereas mutations in the N-terminal half did not display severe secretory deficits. Thus, structural length requirements, largely confined to the C-terminal half of the VAMP2 TMD, seem to be essential for SNARE-mediated membrane-fusion events in cells.


A double-blind randomized comparison of the effect and tolerance of Varidase versus saline when instilled in the urinary bladder in patients with catheter problems.
Thirty-eight patients with problems with their urinary catheters were randomly treated with Varidase or normal saline solution once a day for 10 days. Photographs were taken by using a cystoscope before treatment, at day 10 and day 24 after the commencement of treatment. The photographs were evaluated according to the occurrence of pus, debris, coagula and rubor. In the Varidase group the improvements from baseline to day 10 and day 24 were statistically significant (p less than 0.05) in all parameters while there were no statistically significant improvements in the saline group. Comparing the two treatment groups, regarding the changes from baseline to day 10 and day 24, it was statistically demonstrated that Varidase was significantly more effective than saline in all parameters (p less than 0.05). In conclusion, this study shows that Varidase effectively cleans the urinary bladder from pus, fibrin, necrotic tissue and blood clots.


Adverse Impact of Intraoperative Conversion on the Postoperative Course Following Laparoscopic Pancreaticoduodenectomy.
The aim of the current study was to evaluate the adverse clinical impact of intraoperative conversion during laparoscopic pancreaticoduodenectomy (LPD). The medical records of patients who underwent pancreaticoduodenectomy (PD) were retrospectively reviewed. Perioperative clinical variables were compared between patients who underwent converted PD (cPD) and initially planned open PD (OPD) to investigate the clinical impact and predictive factors of intraoperative conversion during LPD. A total of 171 patients were included. Among them, 31 patients (19.3%) were found to have intraoperative conversion during LPD. Failure of progression due to severe adhesion (12 patients, 7%) and major vessel invasion (7 patients, 4%) were the two most frequent reasons for conversion. On multivariate analysis, age [Exp(β)=1.044, p=0.044] and pancreatic texture [Expa(β)=2.431, p=0.039) were found to be independent factors for predicting intraoperative conversion during LPD. In comparative analysis with the OPD group, the cPD group had a longer operation time (516.8 min vs. 449.9 min, p=0.001), higher rate of postoperative hemorrhage (12.1% vs. 0.85%, p=0.008), higher reoperation rate (9.1% vs. 0%, p=0.01), and higher cost (21886.4 USD vs. 17168.9 USD, p=0.018). Intraoperative conversion during LPD can have an adverse clinical impact on the postoperative course following LPD. Appropriate patients selection and improvement of surgical techniques will be crucial for unnecessary intraoperative conversion and safe LPD.


The BAMSE project: presentation of a prospective longitudinal birth cohort study.
The aims of this prospective and longitudinal project are to establish crucial risk factors for asthma and other allergic diseases in childhood, and to study factors of importance for prognosis at already established allergic disease. Socio-economic factors, such as inequality in health, are also to be addressed. The project started in February 1994. To reach sufficient power, 4,000 children had to be included. In November 1996, this number was reached (4,093). Inclusion in the study was made at 3-4 months of age. At that time, and before induction of allergic disease/ asthma of the child, a questionnaire focused on exposure, genetics and socio-economic factors was answered. Settled dust was sampled for later analysis of furred animal and mite allergens. When the children were aged both 1 and 2 years, their parents were asked to fill in new questionnaires focusing on respiratory and allergic (skin, gastrointestinal) symptoms, but also key variables of exposure. Cases with asthma are identified and, for every case, two matched controls drawn. During the following winter, the homes of cases and controls were investigated and the temperature, indoor humidity, air change rate and NO2 measured. Two hundred cases (5%) were expected to be identified during the first 2 years of the children's lives. Some 479 homes have now been investigated and 97.7% of the original 4,093 children still remain in the cohort. The 2-year symptom follow-up ended in November 1998. The 4-year follow-up started on 1 September 1998 and was planned to be finished in June 2000. Questionnaires (allergic and respiratory symptoms, key variables of exposure at home and day care) are sent out to all 4,093 families. All children are invited for examination, lung function tests (PEF, flow-volume, MVV and oxygen clearance) and physical performance. Blood is taken from all children (20 ml). Allergy screening is performed and specific IgE examined. Blood cells will be frozen to allow for later DNA extraction. In subsets (children with any allergic and/or respiratory manifestation and controls), markers of inflammation in blood and urine will be examined, as well as eosinophils in nasal smear. Interviews are carried out to assess the severity of asthma, type/periodicity of health care given, asthma medication and parental sick leave when appropriate. As a separate project, financed by the EU, outdoor pollution as risk factors for asthma and allergies are to be studied within the BAMSE cohort. A follow-up of 8-9 years is underway.


The cone electroretinogram in retinopathy of prematurity.
To test the hypothesis that retinopathy of prematurity (ROP) affects the cone photoreceptors less than the rod photoreceptors. Electroretinogram (ERG) responses to a 1.8-log-unit range of red flashes on a white, rod-saturating background were recorded in 42 subjects with a history of preterm birth and ROP (28 untreated; 6 treated) or no ROP (n = 8). The sensitivity (S(CONE)) and saturated amplitude (R(CONE)) of the cone photoresponse were calculated by fit of a model of the activation of cone phototransduction to the a-waves. The cone-driven b-wave amplitude was evaluated as a function of stimulus intensity. S(CONE) and R(CONE) were compared to the rod response parameters (S(ROD), R(ROD)) recorded from the same preterm subjects. Responses in the former preterm subjects were compared to those in control subjects. The values of S(CONE) and R(CONE) in the preterm subjects overlapped broadly with those in the control subjects. The shapes of the b-wave stimulus-response functions did not differ between preterm and control subjects. The relative value of S(CONE) was significantly greater than that of S(ROD). ROP has less effect on the cone than on the rod photoresponses, suggesting that cones are more resistant to the ROP disease process. The similar shape of the b-wave stimulus-response function in preterms and control subjects is evidence that ROP does not alter the balance of ON and OFF signals in the cone pathway.


Diltiazem and propranolol, alone and in combination, on exercise performance and left ventricular function in patients with stable effort angina: a double-blind, randomized, and placebo-controlled study.
Diltiazem and propranolol alone and in combination as antianginal agents were compared with placebo in 12 patients with stable exertional angina at Stanford University Medical Center. The patients performed symptom-limited, multi-stage upright bicycle ergometric exercise while undergoing radionuclide angiographic studies every two weeks while being treated with 90 mg of diltiazem four times daily, 60 mg of propranolol four times daily, a combination of 90 mg of diltiazem and 60 mg of propranolol four times daily, and placebo. Diltiazem, propranolol and a combination all significantly increased exercise duration compared to placebo (526 +/- 149, 525 +/- 115, and 549 +/- 129 vs 430 +/- 132 sec.). Although rate pressure product and heart rate decreased with diltiazem therapy at submaximal workloads, these values were unchanged at peak exercise in contrast to propranolol or the combination of propranolol or diltiazem. Diltiazem decreased the sub-maximal and maximal degree of exercise-induced ST segment depression by over 50% compared to placebo (P less than 0.01 vs placebo). Diltiazem resulted in a higher exercise left ventricular ejection fraction compared to placebo, propranolol or the combination of diltiazem or propranolol (all less than P less than 0.05). Sinus bradycardia or orthostatic hypertension occurred in four patients on the high-dose combination therapy and required dose reduction. We concluded that high-dose diltiazem, appeared to be even more effective than moderate-dose propranolol or the combination of diltiazem and propranolol in improving exercise tolerance, electrocardiographic evidence of myocardial ischaemia and left ventricular function in patients with stable effort angina due to occlusive coronary artery disease.


[Molecular characterization and genetic divergence of Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae) on different crops and growing areas].
Knowledge on the genetic variation of populations of Bemisia tabaci (Genn.) can improve the understanding of genetic diversity found in their biotypes and, consequently, offer guidelines for its management. In this study, the molecular characterization was performed and genetic diversity data were obtained for this insect from three regions of Brazil on different crops [cotton and soybean (Mato Grosso - MT); cabbage (Distrito Federal - DF); soybean and potato (São Paulo - SP)], using RAPD markers. RAPD analysis indicated 80.6% polymorphic loci and the average genetic similarity obtained by the Jaccard coefficient was 0.67. The whitefly populations collected on potato (SP) and soybean (MT) had higher genetic diversity values (0.75 and 0.72, respectively). Shannon's index (Ho) showed higher values for potato and soybean (SP e MT), and a smaller value for cabbage (DF). A high genetic divergence within and among the collected populations occurred, structured according to the regions of collection. Moreover, the great genetic similarity observed between potato (SP) and soybean (SP) populations suggested that both belong to the same biotype B and reinforces the polyphagous behavior of the species.


Imaging the pericardium.
Noninvasive imaging has profoundly altered our ability to diagnose and manage patients with suspected pericardial disease. Although pericardial involvement may first become apparent on a chest radiography, echocardiography (M-mode, 2-D, and Doppler) is the procedure of choice for identifying and characterizing pericardial disease. CT and MRI frequently provide complementary information and in certain instances are necessary for diagnosis. Studies that compare diagnostic accuracy in subsets of patients with pericardial disease are clearly needed, but availability, safety, and cost considerations will influence the choice of procedure. Regardless of the imaging study selected, it is imperative that the results be interpreted in the context of the clinical examination.


Human telomerase reverse transcriptase mRNA expression assessed by real-time reverse transcription polymerase chain reaction predicts chemosensitivity in patients with ovarian carcinoma.
To evaluate in vivo whether the expression of the human telomerase reverse transcriptase (hTERT) gene, the catalytic subunit of the telomerase complex, is predictive of response to chemotherapy in ovarian cancer patients. Fifty-nine advanced-stage ovarian cancer patients who were treated with platinum-based chemotherapy were studied. hTERT levels were evaluated by real-time reverse transcriptase polymerase chain reaction (RT-PCR) on tumor specimens obtained before the treatment. Variables were analyzed by the chi(2) and Fisher's exact tests. Logistic regression analysis was also performed to account for the effects of all the covariates investigated (residual disease, stage, histotype, and grade). Twenty-eight (47%) of the 59 tumors showed low hTERT levels, whereas 31 (53%) tumors displayed high hTERT levels. Seventy-five percent of complete responders showed high levels of hTERT expression, whereas 66% of partial responders or nonresponders exhibited low hTERT levels (P =.002). Only residual disease and hTERT expression were independent predictors of response (odds ratios, 13.455 and 7.586, respectively). The combination of these two parameters provides powerful predictive information: 18 of the 20 patients with residual disease more than 2 cm and low hTERT levels were partial responders or nonresponders, whereas 11 of the 12 patients with residual disease less than 2 cm and high hTERT levels showed a complete response (chi(2) = 21,416; P <.00001). Our data indicate that hTERT expression, measured by real-time RT-PCR, is a possible independent marker of response to platinum-based therapy in advanced stage ovarian cancer patients. Prospective validation of this marker will be required to further define its predictive value.


Role of free radicals and antioxidants in nasal polyps.
The aim of this study is to determine the role of free radicals and antioxidants in nasal polyps. Prospective, randomized, controlled study. Thirty-one patients with nasal polyposis and a control group consisting of 19 patients with septal deviation and lower turbinate hypertrophy were included in the study. Levels of the antioxidants retinol, beta-carotene, alpha-tocopherol, and ascorbic acid were measured from the sera of the patients with nasal polyposis and the control group. Plasma levels of superoxide dismutase activity (SOD), glutathione peroxidase (GSHPX) activity, and reduced glutathione (GSH) were also obtained. As a peroxidation product, the levels of the malondialdehyde-thiobarbituric acid (MDA) combination were measured from the plasma of patient and control groups. Measurements of MDA, GSH, and alpha-tocopherol levels were also taken from the polyp tissue and turbinate mucosa of the control group. The blood levels of antioxidants and MDA as an oxidant were significantly different in the patient group compared with the control group (P <.01). The tissue levels of antioxidants and MDA were significantly different in the patients with polyposis compared with the control group (P <.01). The blood and tissue anti-oxidant levels were found to be decreased, and MDA levels as an oxidant increased significantly in the patient group with polyposis when compared with the control group, and there was a negative correlation between oxidative stress and antioxidants. This study demonstrates that oxidative stress and tissue and blood antioxidants in the patients with polyposis were significantly different compared with the control group. The blood and tissue antioxidant levels decreased, and MDA levels, as an oxidant, increased significantly in the patient group with polyposis when compared with the control group. The current study demonstrates that there is strong evidence related to oxidative stress in the pathogenesis of nasal polyposis, and antioxidants can have a preventive role in free-radical-mediated tissue damage in nasal polyposis.


An Approach to the Estimation of Chronic Air Pollution Effects Using Spatio-Temporal Information.
There is substantial observational evidence that long-term exposure to particulate air pollution is associated with premature death in urban populations. Estimates of the magnitude of these effects derive largely from cross-sectional comparisons of adjusted mortality rates among cities with varying pollution levels. Such estimates are potentially confounded by other differences among the populations correlated with air pollution, for example, socioeconomic factors. An alternative approach is to study covariation of particulate matter and mortality across time within a city, as has been done in investigations of short-term exposures. In either event, observational studies like these are subject to confounding by unmeasured variables. Therefore the ability to detect such confounding and to derive estimates less affected by confounding are a high priority. In this article, we describe and apply a method of decomposing the exposure variable into components with variation at distinct temporal, spatial, and time by space scales, here focusing on the components involving time. Starting from a proportional hazard model, we derive a Poisson regression model and estimate two regression coefficients: the "global" coefficient that measures the association between national trends in pollution and mortality; and the "local" coefficient, derived from space by time variation, that measures the association between location-specific trends in pollution and mortality adjusted by the national trends. Absent unmeasured confounders and given valid model assumptions, the scale-specific coefficients should be similar; substantial differences in these coefficients constitute a basis for questioning the model. We derive a backfitting algorithm to fit our model to very large spatio-temporal datasets. We apply our methods to the Medicare Cohort Air Pollution Study (MCAPS), which includes individual-level information on time of death and age on a population of 18.2 million for the period 2000-2006. Results based on the global coefficient indicate a large increase in the national life expectancy for reductions in the yearly national average of PM2.5. However, this coefficient based on national trends in PM2.5 and mortality is likely to be confounded by other variables trending on the national level. Confounding of the local coefficient by unmeasured factors is less likely, although it cannot be ruled out. Based on the local coefficient alone, we are not able to demonstrate any change in life expectancy for a reduction in PM2.5. We use additional survey data available for a subset of the data to investigate sensitivity of results to the inclusion of additional covariates, but both coefficients remain largely unchanged.


Use of a collagen-glycosaminoglycan copolymer (Integra) in combination with adjuvant treatments for reconstruction of severe chest keloids.
Keloids occurring on the chest can be deforming with significant painful sequelae for patients. These lesions can pose a therapeutic dilemma for the dermatologic surgeon as certain excision defects may be too large to close primarily and recurrences tend to be high (40-100%). A collagen-glycosaminoglycan copolymer (Integra) has been found to be useful in the surgical treatment of scar excisions as the bovine collagen and glycosaminoglycans provide a template for neocollagenesis. Additionally, this dermal regeneration template concomitantly reduces tensile forces on the wound. The authors' group has followed five patients with chronic chest keloids refractory to myriad of interventions and treated these patients with surgical excision followed by Integra placement into the wound bed on the chest. Split-thickness grafts were applied shortly thereafter. Patients were followed at regular intervals and all patients received adjuvant therapy with single-dose radiation and intralesional chemotherapy (triamcinolone and/or 5 fluorouracil). This treatment protocol has provided a cure rate of 100% over an average of 43 months follow-up. The symptoms often accompanied by these chest keloids also appear to improve. The authors believe that this study provides the groundwork for further investigation of Integra for surgical management of keloids. A placebo-controlled study should be performed to adequately determine if this data holds true.


The bully within and without: strategies to address horizontal violence in nursing.
Pervasive horizontal violence in the nursing profession has bred the aphorism "nurses eat their young." Nurses must make a concerted effort to advocate for change in the existing professional culture. Such change can begin with embracing helpful tips in overcoming bullying and modeling successful positive behaviors. This paper defines horizontal violence in nursing and explores strategies to address it based upon shared information. Multiple workshops across the country have allowed nurses to exchange information about their experiences with horizontal violence. Nurses have examined their roles as bystander, perpetrator, and victim in discussing contemporary perspectives, employing methods of engagement, promoting debate, and using practical resource advice in dealing with the experience and prevention of horizontal violence. Workshops, evaluated through pretests/posttests and written evaluations, revealed increased knowledge about concepts of horizontal violence and their application to the workplace, recognition of horizontal violence, and methods of dealing with horizontal violence in nursing. Across the profession, nurses need exposure to practical approaches to curtailing horizontal violence in nursing. A similar workshop approach could be adapted across educational programs and workplaces based on the individual needs of each environment.


Tissue-specific cadmium and metallothionein levels in rainbow trout chronically acclimated to waterborne or dietary cadmium.
Rainbow trout were exposed to a sublethal concentration of waterborne Cd (0 or 3 microg/L) or dietary Cd (0 or 500 mg/kg dry wt) for 30 days to induce acclimation, and tissue Cd and metallothionein (MT) levels were examined. The greatest Cd concentrations were observed in the kidney followed by the gills and liver of the fish exposed to Cd via water, but in the gut tissues followed by the kidney, liver, and gills for dietary-exposed fish, reflecting a variation depending on the route of Cd exposure. Some MT was found in the nonacclimated naïve fish with no experience of elevated Cd exposure, and these background MT levels were quite high in the posterior intestine (480 microg/g), cecae (257 microg/g), and liver (248 microg/g) relative to other tissues (7-50 microg/g). With exposure to both waterborne and dietary Cd, MT levels rose significantly in all observed tissues. The increases relative to the control levels of MT in naïve fish were in the order: kidney (5.4 times) > gills (4.6) > liver (1.3) for the waterborne exposure group, and in the order kidney (19.3 times) >> cecae and posterior intestine (approximately 6.5 times) > liver and stomach (approximately 5 times) > midintestine (4.3 times) > gills (2.1 times) for the dietary exposure group. At 24 hours after an acute gastrointestinal dose of Cd (276 microg/kg) infused into the stomach of dietary exposure groups, large increases of total Cd but not MT levels were found in the gut tissues of nonacclimated fish; in the Cd-acclimated fish, the posterior intestine was greatly affected with decreases in Cd (71%), Zn (33%), Cu (70%) and MT (46%) levels, suggesting an enhanced sloughing of tissue materials after infusion. Exposure to Cd did not cause any notable decrease of Zn or Cu in any tissue, except that found in the posterior intestine. However, a molar analysis indicated that although Cd levels remained less than MT binding capacity in both waterborne and dietary exposure groups, the total metal levels (Cd + Zn + Cu) greatly exceeded MT binding capacity in all tissues of Cd-exposed fish, suggesting a potential competition of Cd with other metals for binding sites on MT and non-MT proteins in the tissues.


Feeding cell development by cyst and root-knot nematodes involves a similar early, local and transient activation of a specific auxin-inducible promoter element.
SUMMARY To study the role of the phytohormone auxin in nematode feeding cell induction and early development, the transcriptional regulation of the artificial auxin-responsive promoter element DR5 was monitored in Arabidopsis thaliana roots infected with the cyst nematode Heterodera schachtii or the root-knot nematode Meloidogyne incognita. For both nematode species, a specific and strong activation of DR5::gusA was observed inside the initial feeding cells at 18 h post inoculation, pointing to an increase in the perceived auxin concentration. This high expression was maintained until 3-5 days post inoculation and subsequently the GUS staining was reduced. Cyst and root-knot nematodes are distantly related and the feeding sites they induce are highly dissimilar. In this respect, the similarities between the two nematode-induced DR5 activation patterns in A. thaliana roots are remarkable. A transient and local increase in auxin perception could be due to an accumulation or to an increased sensitivity. Based on previously published data, a local auxin accumulation seems to be the more probable explanation. The observed early and localized increase of the perceived IAA concentration in the initial feeding structure underlines that this phytohormone could be an important clue in feeding cell induction by plant parasitic nematodes.


Early Stages of Alzheimer's Disease: Evolving the Care Team for Optimal Patient Management.
Alzheimer's disease (AD) is a progressive, neurodegenerative disease that creates complex challenges and a significant burden for patients and caregivers. Although underlying pathological changes due to AD may be detected in research studies decades prior to symptom onset, many patients in the early stages of AD remain undiagnosed in clinical practice. Increasing evidence points to the importance of an early and accurate AD diagnosis to optimize outcomes for patients and their families, yet many barriers remain along the diagnostic journey. Through a series of international working group meetings, a diverse group of experts contributed their perspectives to create a blueprint for a patient-centered diagnostic journey for individuals in the early stages of AD and an evolving, transdisciplinary care team. Here, we discuss key learnings, implications, and recommendations.


The Effect of Sheep and Cow Milk Supplementation of a Low Calcium Diet on the Distribution of Macro and Trace Minerals in the Organs of Weanling Rats.
The aim of this study was to investigate the effect of either sheep or cow milk supplementation to a low calcium and phosphorus diet on growth and organ mineral distribution in weanling rats. Rats were fed diets consisting of either a control chow, a 50% reduced calcium and phosphorous chow (low Ca/P), low Ca/P and sheep milk, or low Ca/P and cow milk diet for 28 days. Food intake of the rats, the growth rate of the rats, and the concentrations of minerals in the soft organs and serum were determined. Rats fed the low Ca/P diet alone had lower weight gain than rats consuming either of the milk-supplemented diets (p < 0.05). Both sheep milk and cow milk supplementation overcame the effects of consuming a diet restricted in calcium and phosphorus but the sheep milk was effective at a significantly lower level of milk intake (p < 0.05). Significant differences (p < 0.05) in essential and trace mineral concentrations due to milk type were observed in the kidney, spleen, and liver. For non-essential minerals, significant differences (p < 0.05), related to diet, were observed in all organs for arsenic, cesium, rubidium, and strontium concentrations.


Validity of predictive equations for 24-h urinary potassium excretion based on timing of spot urine collection among adults: the MESA and CARDIA Urinary Sodium Study and NHANES Urinary Sodium Calibration Study.
24-h urine collections are the suggested method to measure daily urinary potassium excretion (uK) but are costly and burdensome to implement. This study tested how well existing equations with the use of spot urine samples can estimate 24-h uK and if accuracy varies by timing of spot urine collection, age, race, or sex. This cross-sectional study used data from 407 participants aged 18-39 y from the Washington, DC area in 2011 and 554 participants aged 45-79 y from Chicago in 2013. Spot urine samples were collected in individual containers for 24 h, and 1 for each timed period (morning, afternoon, evening, and overnight) was selected. For each selected timed spot urine, 24-h uK was predicted through the use of published equations. Difference (bias) between predicted and measured 24-h uK was calculated for each timed period and within age, race, and sex subgroups. Individual-level differences were assessed through the use of Bland-Altman plots and correlation tests. For all equations, regardless of the timing of spot urine, mean bias was usually significantly different than 0. No one prediction equation was unbiased across all sex, race, and age subgroups. With the use of the Kawasaki and Tanaka equations, 24-h uK was overestimated at low levels and underestimated at high levels, whereas observed differential bias with the Mage equation was in the opposite direction. Depending on prediction equation and timing of urine sample, 61-75% of individual 24-h uKs were misclassified among 500-mg incremental categories from <1500 to ≥3000 mg. Correlations between predicted and measured 24-h uK were poor to moderate (0.19-0.71). Because predicted 24-h uK accuracy varies by timing of spot urine collection, published prediction equations, and within age-race-sex subgroups, study results making use of predicted 24-h uK in association with health outcomes should be interpreted with caution. It is possible that a more accurate prediction equation can be developed leading to different results.


A method for evaluating dynamic functional network connectivity and task-modulation: application to schizophrenia.
In this paper, we develop a dynamic functional network connectivity (FNC) analysis approach using correlations between windowed time-courses of different brain networks (components) estimated via spatial independent component analysis (sICA). We apply the developed method to fMRI data to evaluate it and to study task-modulation of functional connections. We study the theoretical basis of the approach, perform a simulation analysis and apply it to fMRI data from schizophrenia patients (SP) and healthy controls (HC). Analyses on the fMRI data include: (a) group sICA to determine regions of significant task-related activity, (b) static and dynamic FNC analysis among these networks by using maximal lagged-correlation and time-frequency analysis, and (c) HC-SP group differences in functional network connections and in task-modulation of these connections. This new approach enables an assessment of task-modulation of connectivity and identifies meaningful inter-component linkages and differences between the two study groups during performance of an auditory oddball task (AOT). The static FNC results revealed that connectivities involving medial visual-frontal, medial temporal-medial visual, parietal-medial temporal, parietal-medial visual and medial temporal-anterior temporal were significantly greater in HC, whereas only the right lateral fronto-parietal (RLFP)-orbitofrontal connection was significantly greater in SP. The dynamic FNC revealed that task-modulation of motor-frontal, RLFP-medial temporal and posterior default mode (pDM)-parietal connections were significantly greater in SP, and task modulation of orbitofrontal-pDM and medial temporal-frontal connections were significantly greater in HC (all P < 0.05). The task-modulation of dynamic FNC provided findings and differences between the two groups that are consistent with the existing hypothesis that schizophrenia patients show less segregated motor, sensory, cognitive functions and less segregated default mode network activity when engaged with a task. Dynamic FNC, based on sICA, provided additional results which are different than, but complementary to, those of static FNC. For example, it revealed dynamic changes in default mode network connectivities with other regions which were significantly different in schizophrenia in terms of task-modulation, findings which were not possible to discover by static FNC.


Is It Possible to Publish a Calibration Function for Radiochromic Film?
To assess the possibility of using a public calibration function for radiochromic film dosimetry in dose QA of highly conformal treatment plans. EBT3 film calibration strips (3.5 × 20 cm2 from lots A101212 and A011713) were exposed on a Varian Trilogy at a facility to a 10 × 10 cm2 open field at doses of 80, 160, 320 cGy using 6MV photons. Together with a strip of unexposed film from the same lot the exposed films were digitized in a single scan using different Epson 10,000 XL scanners at two different facilities. The dose-response data for each color-channel from each facility were generated using the same calibration function X(D) = a + b/(D - c), where X(D) is the response at dose D and a, b and c are the coefficients. Different batches of EBT3 film were exposed to a VMAT beam. These films, plus two reference strips exposed to doses of zero and 160 cGy, were digitized on the scanners at the two facilities. Using the multi-channel dosimetry method and One-scan protocol (Med Phys, 39:6339-49, 2012) the recorded doses on the VMAT films were calculated and the results were compared with the VMAT plan using a Gamma index of 3%/3 mm. The passing rates obtained for dose maps calculated for all combinations of VMAT images and calibration functions were nearly unchanged, using the One-scan protocol. Also, in all cases a passing rate of >99% was obtained for Gamma index of 3%/3 mm. On the other hand, if the One-scan protocol was not employed, the dose maps for VMAT images and calibration functions from different scanners showed poor correlation with the treatment plan. This is probably due to the scan-to-scan variability. The authors have found that it is feasible to use a public calibration function for a given radiochromic film lot using the same methodology, One-scan protocol, for patient-specific QA.


Engineering enters new cycle of development and definition.
The profession of engineering is evidencing important changes under the impact of the microprocessor and the energy crisis. Societal considerations are altering the normative criteria of engineering while computerized manufacturing systems require entirely new engineering approaches. The trend is toward greater autonomy of engineering from the traditional basic sciences. Significant changes in engineering education will be necessary. The new technological environment requires closer links among academia, industry, and the professional societies-engineering's essential triad.


The role of the CCR1 receptor in the inflammatory response to tobacco smoke in a mouse model.
The aim was to create pathological changes in mice relevant to human smoke exposure that can be used to further understand the mechanisms and pathology of smoke-induced inflammatory disease. Mice were exposed to tobacco smoke or lipopolysaccharide (LPS) to generate an inflammatory infiltrate within the lungs. Tobacco smoke exposure over a 4 day period led to neutrophilia in the lungs of BALB/c mice. Within the inflammatory exudates, significant changes were also seen in protein levels of IL-1B, IL-6, MIP-2, KC (IL-8) and TIMP-1 as measured by ELISA. Further protein changes, as measured via multiplex analysis revealed increased levels of MMP-9, MDC, LIF and MCP-1, amongst other mediators. Major changes in whole lung tissue gene expression patterns were observed. The neutrophilia seen after smoke exposure was steroid-insensitive, relative to doses of steroid needed to reduce LPS-driven neutrophilia in controls. This exposes pathological switches that are changed upon exposure to tobacco smoke, rendering steroids less effective under these conditions. Challenge of chemokine receptor type 1 (CCR1) KO mice in the tobacco smoke model showed that lack of this gene protected the mice from smoke-induced inflammation. This suggests the CCR1 receptor has a key role in the pathogenesis of smoke-induced inflammation.


Detection of Helicobacter pylori DNA in human feces by PCR: DNA stability and removal of inhibitors.
In this study, the stability of Helicobacter pylori DNA in human feces and the effect of a diet lacking in plant material, the suspected source of PCR inhibitors in human feces, were investigated. In addition, a method to remove these inhibitors was developed. Stools inoculated with H. pylori were used as a model. For this purpose, a H. pylori suspension (10(8) CFU/ml) was used to spike stool samples obtained from four healthy adults known to be H. pylori negative. The evaluation of the stability of H. pylori DNA in feces showed that DNA was degraded after 3 days of contact with fecal material at 37 degrees C. A 2-day diet completely free of plant material was sufficient to eliminate PCR inhibitors from human feces. However, inhibitors were detected 48 h after a normal diet was resumed. A new technique consisting of agarose blocks containing embedded DNA as a template for PCR amplification was used for removal of inhibitors, following DNA extraction by a modified QIAamp tissue method (Qiagen, Hilden, Germany). When this method was applied to inhibiting stool samples known to have an inhibitory effect and spiked with H. pylori (5.10(8) CFU/g), a positive PCR was obtained showing that inhibitors present in the original DNA samples were completely removed. The agarose embedded DNA block method is highly efficient and provides clean, high quality template DNA for PCR purposes avoiding long and fastidious conventional extraction methods. In conclusion, this study confirms that H. pylori DNA degrades with time in stools. A diet free of plant material or a special DNA preparation can be used to remove inhibitors and to allow the detection of H. pylori.


miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1.
miR-25, a member of the miR-106b-25 cluster, has been reported as playing an important role in many biological processes by numerous studies, while the role of miR-25 in metabolism and its transcriptional regulation mechanism remain unclear. In this study, gain-of-function and loss-of-function assays demonstrated that miR-25-3p positively regulated the metabolism of C2C12 cells by attenuating phosphoinositide 3-kinase (PI3K) gene expression and triglyceride (TG) content, and enhancing the content of adenosine triphosphate (ATP) and reactive oxygen species (ROS). Furthermore, the results from bioinformatics analysis, dual luciferase assay, site-directed mutagenesis, qRT-PCR, and Western blotting demonstrated that miR-25-3p directly targeted the AKT serine/threonine kinase 1 (Akt1) 3' untranslated region (3'UTR). The core promoter of miR-25-3p was identified, and the transcription factor activator protein-2α (AP-2α) significantly increased the expression of mature miR-25-3p by binding to its core promoter in vivo, as indicated by the chromatin immunoprecipitation (ChIP) assay, and AP-2α binding also downregulated the expression of Akt1. Taken together, our findings suggest that miR-25-3p, positively regulated by the transcription factor AP-2α, enhances C2C12 cell metabolism by targeting the Akt1 gene.


Simplified analysis of pathogenic leptospiral serovars by random amplified polymorphic DNA fingerprinting.
A rapid, simplified procedure combining random amplified polymorphic DNA (RAPD) fingerprinting of boiled cultures with high resolution agarose gel electrophoresis was used to compare strains from 46 pathogenic leptospiral serovars. The serovars were placed in eight groups on the basis of RAPD profile similarities. Groups 1-7 corresponded with the genome species Leptospira interrogans, L. borgpetersenii, L. santarosai, L. noguchii, L. weilii, L. kirschneri and L. meyeri. The eighth group did not correspond with a known genome species and may represent a new genome species. Primer choice determined the degree of discrimination possible between closely related serovars and genotypes. This procedure, unlike other procedures used for analysing taxonomic relationships between leptospiral serovars, does not require extensive DNA purification, polyacrylamide gel electrophoresis or autoradiography.


Reconstitution of pterin-free inducible nitric-oxide synthase.
Inducible nitric-oxide synthase (NOS) was expressed and purified in the absence of 6(R)-tetrahydro-l-biopterin (H(4)B). Pterin-free NOS exhibits a Soret band (416-420 nm) characteristic of predominantly low spin heme and does not catalyze the formation of nitric oxide (. NO) (Rusche, K. M., Spiering, M. M., and Marletta, M. A. (1998) Biochemistry 37, 15503-15512). Reconstitution of pterin-free NOS with H(4)B was monitored by a shift in the Soret band to 396-400 nm, the recovery of.NO-forming activity, and the measurement of H(4)B bound to the enzyme. As assessed by these properties, H(4)B binding was not rapid and required the presence of a reduced thiol. Spectral changes and recovery of activity were incomplete in the absence of reduced thiol. Full reconstitution of holoenzyme activity and stoichiometric H(4)B binding was achieved in the presence of 5 mm glutathione (GSH). Preincubation with GSH before the addition of H(4)B decreased, whereas lower concentrations of GSH extended, the time required for reconstitution. Six protected cysteine residues in pterin-free NOS were identified by labeling of NOS with cysteine-directed reagents before and after reduction with GSH. Heme and metal content of pterin-free and H(4)B-reconstituted NOS were also measured and were found to be independent of H(4)B content. Additionally, pterin-free NOS was reconstituted with 6-methylpterin analogs, including redox-stable deazapterins. Reconstitution with the redox-stable pterin analogs was neither time- nor thiol-dependent. Apparent binding constants were determined for the 6-methyl- (50 microm) and 6-ethoxymethyl (200 microm) deazapterins. The redox-stable pterin analogs appear to bind to NOS in a different manner than H(4)B.


[Use of a questionnaire for screening people with undiagnosed diabetes].
To evaluate a screening questionnaire to identify individuals with undiagnosed diabetes. We conducted a community survey to detect people at increased risk for diabetes using a questionnaire proposed by Herman et al. (1995), that incorporates major risk factors as age, obesity, family history of diabetes, sedentary lifestyle and personal history of delivering a macrosomic infant. Blood glucose test was made by means of reagent strip and a reflectance meter system. We used the ADA recommendations and cut-points for screening programs, adjusted for fasting and random blood glucose. We included 360 participants older than 20 years of age. A total of 200 subjects (55.5%) were at risk for diabetes according to the questionnaire, of whom 31 (15.5%) had an abnormal glucose test compared to the 4.4% of the low-risk group (p < 0.001). The 1995 Herman et al. Questionnaire had sensitivity of 81.6%, specificity of 47.5, positive predictive value of 15.5 and negative predictive value of 95.6%. The high-risk group was older (44.9 vs. 34.6 y, p < 0.001) and heavier (30.5 vs. 24.4 kg/m2, p < 0.001) than the low-risk group. There were 38/360 (10.5%) abnormal glucose readings, of which 31 (81.5%) had a positive questionnaire (p < 0.01). The mean fasting glucose in the high-risk group was higher (90.6 vs. 84.2 mg/dL, p = 0.015) than in those with low risk by questionnaire, as well as for random blood glucose (116.1 vs. 100 mg/dL, p < 0.01). The questionnaire proposed by Herman et al. combined with capillary blood glucose testing performance good in mexican population to identify people at high risk for undiagnosed diabetes, and improved the detection rate.


Kinetic activation-relaxation technique.
We present a detailed description of the kinetic activation-relaxation technique (k-ART), an off-lattice, self-learning kinetic Monte Carlo (KMC) algorithm with on-the-fly event search. Combining a topological classification for local environments and event generation with ART nouveau, an efficient unbiased sampling method for finding transition states, k-ART can be applied to complex materials with atoms in off-lattice positions or with elastic deformations that cannot be handled with standard KMC approaches. In addition to presenting the various elements of the algorithm, we demonstrate the general character of k-ART by applying the algorithm to three challenging systems: self-defect annihilation in c-Si (crystalline silicon), self-interstitial diffusion in Fe, and structural relaxation in a-Si (amorphous silicon).


Efficacy of Calcipotriol-Betamethasone Ointment in Patients with Mild to Moderate Plaque Psoriasis: Subgroup Analyses.
Several factors have been shown to affect psoriasis pathogenesis, clinical presentation and treatment response. The aim of this study was to investigate the potential relationship between patients' baseline characteristics and the efficacy of calcipotriol-betamethasone ointment in patients with mild to moderate plaque psoriasis and to evaluate whether the efficacy is consistent across subgroups. Using data from the therapeutic equivalence study on patients with plaque psoriasis, post hoc analyses were performed to evaluate the impact of baseline demographic and disease characteristics, habits and comorbidities on the response to treatment with calcipotriol-betamethasone ointment. Body mass index (BMI) and obesity were each independently associated (univariate analysis, p < 0.05) with reduction in modified Psoriasis Area and Severity Index (mPASI) and PASI75 (≥75% improvement in mPASI from baseline). Increased body weight is more common in patients with late-onset psoriasis. There was a significant trend for lower response rates with increasing BMI (p = 0.007) and obesity (p = 0.003). The odds of achieving PASI75 is 2.3 times lower for obese compared to normal-weight subjects.If patients with obesity or hypertension were treated with calcipotriol-betamethasone, they were still more likely to achieve PASI75 after 4-week treatment compared to vehicle (p < 0.001). Increased BMI and obesity present risk factors for reduced treatment effectiveness. Importantly, the efficacy of calcipotriol-betamethasone ointment was consistent in all subgroups.


Small heat-shock proteins interact with a flanking domain to suppress polyglutamine aggregation.
Small heat-shock proteins (sHsps) are molecular chaperones that play an important protective role against cellular protein misfolding by interacting with partially unfolded proteins on their off-folding pathway, preventing their aggregation. Polyglutamine (polyQ) repeat expansion leads to the formation of fibrillar protein aggregates and neuronal cell death in nine diseases, including Huntington disease and the spinocerebellar ataxias (SCAs). There is evidence that sHsps have a role in suppression of polyQ-induced neurodegeneration; for example, the sHsp alphaB-crystallin (alphaB-c) has been identified as a suppressor of SCA3 toxicity in a Drosophila model. However, the molecular mechanism for this suppression is unknown. In this study we tested the ability of alphaB-c to suppress the aggregation of a polyQ protein. We found that alphaB-c does not inhibit the formation of SDS-insoluble polyQ fibrils. We further tested the effect of alphaB-c on the aggregation of ataxin-3, a polyQ protein that aggregates via a two-stage aggregation mechanism. The first stage involves association of the N-terminal Josephin domain followed by polyQ-mediated interactions and the formation of SDS-resistant mature fibrils. Our data show that alphaB-c potently inhibits the first stage of ataxin-3 aggregation; however, the second polyQ-dependent stage can still proceed. By using NMR spectroscopy, we have determined that alphaB-c interacts with an extensive region on the surface of the Josephin domain. These data provide an example of a domain/region flanking an amyloidogenic sequence that has a critical role in modulating aggregation of a polypeptide and plays a role in the interaction with molecular chaperones to prevent this aggregation.


Chemistry and biology of cyclotides: circular plant peptides outside the box.
Cyclotides stand out as the largest family of circular proteins of plant origin hitherto known, with more than 280 sequences isolated at peptide level and many more predicted from gene sequences. Their unusual stability resulting from the signature cyclic cystine knot (CCK) motif has triggered a broad interest in these molecules for potential therapeutic and agricultural applications. Since the time of the first cyclotide discovery, our laboratory in Uppsala has been engaged in cyclotide discovery as well as the development of protocols to isolate and characterize these seamless peptides. We have also developed methods to chemically synthesize cyclotides by Fmoc-SPPS, which are useful in protein grafting applications. In this review, experience in cyclotide research over two decades and the recent literature related to their structures, synthesis, and folding as well the recent proof-of-concept findings on their use as "epitope" stabilizing scaffolds are summarized.


Propagation and titration of Alkhumra hemorrhagic fever virus in the brains of newborn Wistar rats.
Alkhumra hemorrhagic fever virus (AHFV) is a novel flavivirus identified first in Saudi Arabia. In this study, successful propagation of AHFV in the brains of newborn Wistar rats is described and the median rat lethal dose (RLD50) is determined. AHFV-RNA-positive human sera diluted 1:10 were injected intracerebrally into 16, ≤24h old rats. Post-inoculation, the rats were observed daily for 30 days. Brains of moribund rats were tested for AHFV-RNA using RT-PCR and cultured in LLC-MK2 cells. The titer of the isolated virus was determined and expressed in median tissue culture infectious dose (TCID50). To determine the RLD50, AHFV brain suspension was 10-fold diluted serially and each dilution was inoculated in the cerebral hemispheres of 10 rats for a total of 90 rats. Three days post-inoculation, the rats developed tremor, irritability, convulsion, opisthotonus, and spastic paresis starting in the hind limbs and ascending to involve the whole body. All infected rats died within 3-7 days with histopathologically confirmed meningoencephalitis. AHFV-RNA was detected in the brains of all infected rats and the virus titer was 10(9.4) RLD50/ml. The virus titer in LLC-MK2 was 10(8.2) TCID50/ml. In conclusion, AHFV was propagated successfully to high titers in the brains of newborn Wistar rats.


Plastid and mitochondrial genomes of Coccophora langsdorfii (Fucales, Phaeophyceae) and the utility of molecular markers.
Coccophora langsdorfii (Turner) Greville (Fucales) is an intertidal brown alga that is endemic to Northeast Asia and increasingly endangered by habitat loss and climate change. We sequenced the complete circular plastid and mitochondrial genomes of C. langsdorfii. The circular plastid genome is 124,450 bp and contains 139 protein-coding, 28 tRNA and 6 rRNA genes. The circular mitochondrial genome is 35,660 bp and contains 38 protein-coding, 25 tRNA and 3 rRNA genes. The structure and gene content of the C. langsdorfii plastid genome is similar to those of other species in the Fucales. The plastid genomes of brown algae in other orders share similar gene content but exhibit large structural recombination. The large in-frame insert in the cox2 gene in the mitochondrial genome of C. langsdorfii is typical of other brown algae. We explored the effect of this insertion on the structure and function of the cox2 protein. We estimated the usefulness of 135 plastid genes and 35 mitochondrial genes for developing molecular markers. This study shows that 29 organellar genes will prove efficient for resolving brown algal phylogeny. In addition, we propose a new molecular marker suitable for the study of intraspecific genetic diversity that should be tested in a large survey of populations of C. langsdorfii.


The potential of amniotic membrane/amnion-derived cells for regeneration of various tissues.
Regenerative medicine is a new field based on the use of stem cells to generate biological substitutes and improve tissue functions, restoring damaged tissue with high proliferability and differentiability. It is of interest as a potential alternative to complicated tissue/organ transplantation. Recently, amnion-derived cells have been reported to have multipotent differentiation ability, and these cells have attracted attention as a cell source for cell-transplantation therapy. The amnion possesses considerable advantageous characteristics: the isolated cells can differentiate into all three germ layers; they have low immunogenicity and anti-inflammatory functions; and they do not require the sacrifice of human embryos for their isolation, thus avoiding the current controversies associated with the use of human embryonic stem cells. Moreover, we developed human amniotic cell-sheets using a novel culture surface coated with a noncytotoxic, temperature-responsive elastic protein-based polymer. We also generated a "hyper-dry-amnion", which has already been applied clinically in the ophthalmological field. Compared to cryopreserved fresh amnion, "hyper-dry-amnion" is easy to handle and has started to bring good results to patients. These materials from the amnion are also expected to open a new field in tissue engineering. Thus, amnion, which had been discarded after parturition, has started to be appreciated as an attractive material in the field of regenerative medicine. In this review, the most recent and relevant clinical and experimental data about the use of amniotic membrane and cells derived from it are described.


Tissue Responses to Shiga Toxin in Human Intestinal Organoids.
Shiga toxin (Stx)-producing Escherichia coli (eg, O157:H7) infection produces bloody diarrhea, while Stx inhibits protein synthesis and causes the life-threatening systemic complication of hemolytic uremic syndrome. The murine intestinal tract is resistant to O157:H7 and Stx, and human cells in culture fail to model the complex tissue responses to intestinal injury. We used genetically identical, human stem cell-derived intestinal tissues of varying complexity to study Stx toxicity in vitro and in vivo. In vitro susceptibility to apical or basolateral exposure to Stx was assessed using human intestinal organoids (HIOs) derived from embryonic stem cells, or enteroids derived from multipotent intestinal stem cells. HIOs contain a lumen, with a single layer of differentiated epithelium surrounded by mesenchymal cells. Enteroids only contain epithelium. In vivo susceptibility was assessed using HIOs, with or without an enteric nervous system, transplanted into mice. Stx induced necrosis and apoptotic death in both epithelial and mesenchymal cells. Responses that require protein synthesis (cellular proliferation and wound repair) also were observed. Epithelial barrier function was maintained even after epithelial cell death was seen, and apical to basolateral translocation of Stx was seen. Tissue cross-talk, in which mesenchymal cell damage caused epithelial cell damage, was observed. Stx induced mesenchymal expression of the epithelial marker E-cadherin, the initial step in mesenchymal-epithelial transition. In vivo responses of HIO transplants injected with Stx mirrored those seen in vitro. Intestinal tissue responses to protein synthesis inhibition by Stx are complex. Organoid models allow for an unprecedented examination of human tissue responses to a deadly toxin.


[The nosological entity bulimia nerviosa].
Anorexia nervosa and bulimia nervosa are at the present moment, two well defined clinical entities among the group of the eating disorders. The psychopathological differentiation of both syndromes has a great importance for diagnosis and therapy. The authors make a phenomenological description, based on case histories of patients with diagnostics of anorexia and bulimia nervosa, and try to establish an approach to the essential symptomatology of those disorders. The presence of affective symptomatology--depressive, but not exclusively--in the eating behaviour disorders in general and particularly in bulimia nervosa, is nowadays interpreted as an unspecific emotional lability as a response to stressing situations. That is to say, it is a secondary depressive symptomatology, more than a primary mood disorder preceding or underlying bulimia. There is strong evidence in favour of a dysregulation of serotonin metabolism in patients with bulimia nervosa, in the sense of a reduced activity, which manifest itself clinically by binges with food with a high content in carbohydrates. High levels of 5-HT seem to induce increasing feelings of safety, fullness and lead to an interruption of eating. Fluoxetine and this active metabolite are selective inhibitors of the reuptake of 5-HT and their antibulimic effect could be mediated by this mechanism.


Restoration of vision I: neurobiological mechanisms of restoration and plasticity after brain damage - a review.
Lesions in the central nervous system often lead to loss of vision due to visual system involvement. In the course of weeks or months, some vision can recover in rats, cats, monkeys, and humans, though it is mostly incomplete. This paper reviews the current knowledge of the under-lying neurobiological mechanism of recovery of vision, particularly those observed in adult rats with partial optic nerve crush (ONC). Immediately after ONC, rats are almost completely blind, evident by their inability to perform visual tasks such as brightness and pattern discrimination and they fail to orient towards small, moving targets. Within about two weeks, however, rats significantly recover some of their lost visual functions despite the fact that only about 10 % of the retinal ganglion cells (RGCs) maintain a viable connection with their brain tar-gets. Molecular, anatomical and physiological studies have identified some of the neurobiological determinants of this visual restitution. Immediately following ONC there is a massive soma swelling of about 80 % of the RGCs with subsequent cell death due to apoptosis and necrosis. The remaining 20 % survive with or without axonal connection to their target and undergo marked changes: (i) about half of these RGCs experience a moderate, reversible soma swelling, (ii) there is a loss of anterograde axonal transport in the optic nerve which partially recovers after several weeks, and (iii) many of the surviving RGCs undergo alterations in gene expression, particularly that of the NR1 receptor and the immediate early gene, c-jun. While these changes may be part of an adaptive program of the cells to cope with the trauma, cell survival in the retina does not correlate well with subsequent recovery of vision. We have therefore also studied plasticity in the down stream denervated brain structures which are innervated by retinofugal pathways, particularly the superior colliculus. Here we found (i) recovery of metabolic activity and (ii) changes in gene expression, such as an up-regulation of the enhancer of split (R esp-1) gene. When viewed together with studies on recovery of vision and neuronal reorganization done in other laboratories it is clear that recovery of vision involves simultaneous plasticity in the damaged structure itself and, transsynaptically, in down-stream structures such as the tectum, the lateral geniculate nucleus and visual cortex. Considering both pre-clinical and clinical evidence, I propose the biological substrate underlying restoration of vision as follows: surviving neurons within areas of partial damage - which corresponds clinically to "transition zones" located between intact and deficient visual field sectors - act in concert with down-stream areas, which themselves undergo dramatic reorganization to cope with a condition of reduced, but residual input. Restitution of vision is therefore a multifactorial event of within-systems plasticity, involving neurobiological alterations along the entire retinofugal axis. It is likely that these mechanisms are also responsible for visual improvements that are seen both in animals and patients after prolonged visual restitution training.


Adaptive mammographic image enhancement using first derivative and local statistics.
This paper proposes an adaptive image enhancement method for mammographic images, which is based on the first derivative and the local statistics. The adaptive enhancement method consists of three processing steps. The first step is to remove the film artifacts which may be misread as microcalcifications. The second step is to compute the gradient images by using the first derivative operators. The third step is to enhance the important features of the mammographic image by adding the adaptively weighted gradient images. Local statistics of the image are utilized for adaptive realization of the enhancement, so that image details can be enhanced and image noises can be suppressed. The objective performances of the proposed method were compared with those by the conventional image enhancement methods for a simulated image and the seven mammographic images containing real microcalcifications. The performance of the proposed method was also evaluated by means of the receiver operating-characteristics (ROC) analysis for 78 real mammographic images with and without microcalcifications.


[Dynamics of morpho-functional changes in gastrin-producing cells of the stomach after electric stimulation of the vagus nerve].
In 25 white rats, direct stimulation of the vagus nerve increased the average diameter of secretory granules and decreased their number and volume, thus reflecting the release of gastrin during first 5 min after the stimulation. Subsequent increase in volume of nucleus, nucleolus, endoplasmatic reticulum, mitochondria and Golgi complex suggest rapid activation of hormone synthesis in G cells under vagostimulation. The data obtained allow to revise the existing concepts of gastrin release from cells and of ultrastructural criteria of this process.


Hb lepore/β0-thalassaemia with α+-thalassaemia interactions, a potential diagnostic pitfall.
Haemoglobin (Hb) Lepore is a variant Hb consisting of two α-globin and two δβ-globin chains. In a heterozygote, it is associated with clinical findings of thalassaemia minor, but interactions with other haemoglobinopathies can lead to various clinical phenotypes and pose diagnostic challenges. We reported a pair of siblings from a Malay family, who presented with pallor and hepatosplenomegaly at the ages of 21 months and 14 months old. The red cell indices and peripheral blood smears of both patients showed features of thalassaemia intermedia. Other laboratory investigations of the patients showed conflicting results. However, laboratory investigation results of the parents had led to a presumptive diagnosis of compound heterozygote Hb Lepore/β-thalassaemia and co-inheritance α+-thalassaemia (-α3.7). Hb Lepore has rarely been detected in Southeast Asian countries, particularly in Malaysia. These two cases highlight the importance of family studies for accurate diagnosis, hence appropriate clinical management and genetic counseling.


Superoxide anion generation by the cytochrome bc1 complex.
We have measured the rates of superoxide anion generation by cytochrome bc(1) complexes isolated from bovine heart and yeast mitochondria and by cytochrome bc(1) complexes from yeast mutants in which the midpoint potentials of the cytochrome b hemes and the Rieske iron-sulfur cluster were altered by mutations in those proteins. With all of the bc(1) complexes the rate of superoxide anion production was greatest in the absence of bc(1) inhibitor and ranged from 3% to 5% of the rate of cytochrome c reduction. Stigmatellin, an inhibitor that binds to the ubiquinol oxidation site in the bc(1) complex, eliminated superoxide anion formation, while myxothiazol, another inhibitor of ubiquinol oxidation, allowed superoxide anion formation at a low rate. Antimycin, an inhibitor that binds to the ubiquinone reduction site in the bc(1) complex, also allowed superoxide anion formation and at a slightly greater rate than myxothiazol. Changes in the midpoint potentials of the cytochrome b hemes had no significant effect on the rate of cytochrome c reduction and only a small effect on the rate of superoxide anion formation. A mutation in the Rieske iron-sulfur protein that lowers its midpoint potential from +285 to +220 mV caused the rate of superoxide anion to decline in parallel with a decline in cytochrome c reductase activity. These results indicate that superoxide anion is formed by similar mechanisms in mammalian and yeast bc(1) complexes. The results also show that changes in the midpoint potentials of the redox components that accept electrons during ubiquinol oxidation have only small effects on the formation of superoxide anion, except to the extent that they affect the activity of the enzyme.