Internalization of bacterial redox protein azurin in mammalian cells: entry domain and specificity.
Azurin is a member of a group of copper-containing redox proteins called cupredoxins. Different cupredoxins are produced by different aerobic bacteria as agents of electron transfer. Recently, we demonstrated that azurin enters into J774 and several types of cancer cells leading to the induction of apoptosis. We now demonstrate that azurin is internalized in J774 or cancer cells in a temperature-dependent manner. Azurin shows preferential entry into cancer compared with normal cells. An 28-amino-acid fragment of azurin fused to glutathione S-transferase (GST) or the green fluorescent protein (GFP), which are incapable of entering mammalian cells by themselves, can be internalized in J774 or human melanoma or breast cancer cells at 37 degrees C, but not at 4 degrees C. Competition experiments as well as studies with inhibitors such as cytochalasin D suggest that azurin may enter cells, at least in part, by a receptor-mediated endocytic process. The 28-amino-acid peptide therefore acts as a potential protein transduction domain (PTD), and can be used as a vehicle to transport cargo proteins such as GST and GST-GFP fusion proteins. Another member of the cupredoxin family, rusticyanin, that has also been shown to enter J774 and human cancer cells and exert cytotoxicity, does not demonstrate preferential entry for cancer cells and lacks the structural features characteristic of the azurin PTD.


The impact of antimicrobial photodynamic therapy in an artificial biofilm model.
The susceptibility of bacterial cultures in biofilm formations is important for a variety of clinical treatment procedures. Therefore, the aim of the study was to assess the impact of laser-induced antimicrobial photodynamic therapy on the viability of Streptococcus mutans cells employing an artificial biofilm model. Using sterile chambered coverglasses, a salivary pellicle layer was formed in 40 chambers. Streptococcus mutans cells were inoculated in a sterile culture medium. Employing a live/dead bacterial viability kit, bacteria with intact cell membranes stained fluorescent green. Each pellicle-coated test chamber was filled with 0.7 ml of the bacterial suspension and analysed using a confocal laser scanning microscope within a layer of 10 μm at intervals of 1 μm from the pellicle layer. Phenothiazine chloride was used as a photosensitizer in all 40 test chambers. A diode laser (wavelength 660 nm, output power 100 mW) was used to irradiated 20 chambers for 2 min. Fluorescence values in the test chambers after laser irradiation (median 2.1 U, range 0.4-3.4 U) were significantly lower than baseline values after adding the photosensitizer (median 3.6 U, range 1.1-9.0; p < 0.05). The non-irradiated control chambers showed no change in fluorescence at the end of an additional photosensitizer residence time of 2 min without laser irradiation (median 1.9 U, range 0.7-3.6; median 1.9 U, range 0.8-6.0, respectively; p > 0.05). The present study indicated that laser irradiation is an essential part of antimicrobial photodynamic therapy to reduce bacteria within a layer of 10 μm. Further studies are needed to evaluate the maximum biofilm thickness that still allows a toxic effect on microorganisms.


Pregnancies complicated by gestational diabetes and fetal growth restriction: an analysis of maternal and fetal body composition using magnetic resonance imaging.
Maternal body composition may influence fetal body composition. The objective of this pilot study was to investigate the relationship between maternal and fetal body composition. Three pregnant women cohorts were studied: healthy, gestational diabetes (GDM), and fetal growth restriction (FGR). Maternal body composition (visceral adipose tissue volume (VAT), subcutaneous adipose tissue volume (SAT), pancreatic and hepatic proton-density fat fraction (PDFF) and fetal body composition (abdominal SAT and hepatic PDFF) were measured using MRI between 30 to 36 weeks gestation. Compared to healthy and FGR fetuses, GDM fetuses had greater hepatic PDFF (5.2 [4.2, 5.5]% vs. 3.2 [3, 3.3]% vs. 1.9 [1.4, 3.7]%, p = 0.004). Fetal hepatic PDFF was associated with maternal SAT (r = 0.47, p = 0.02), VAT (r = 0.62, p = 0.002), and pancreatic PDFF (r = 0.54, p = 0.008). When controlling for maternal SAT, GDM increased fetal hepatic PDFF by 0.9 ([0.51, 1.3], p = 0.001). In this study, maternal SAT, VAT, and GDM status were positively associated with fetal hepatic PDFF.


Early effect of progesterone on levels of cyclic adenosine 3':5'-monophosphate in Xenopus oocytes.
Progesterone treatment of Xenopus oocytes in vitro causes progression through meiotic cell division. The role of altered intracellular levels of cAMP on the initiation of meiotic cell division has been studied. Basal levels of cAMP averaged 1.5 pmol in oocytes from eight females, and exposure to progesterone caused a rapid drop in cAMP to about 40 to 60% of basal. Half-maximal decreases occurred within 15 to 60 s, and cAMP returned to near basal values by 20 min after progesterone. Theophylline inhibition of progesterone-induced cell division was characterized by a small increase in basal levels of cAMP and a reduced drop in cAMP due to the hormone. Cholera toxin, an activator of adenylate cyclase, was found to be a potent inhibitor of progesterone-induced meiosis, with half-maximal inhibition at 8 times 10(-12) M. In addition, the purified A subunit of cholera toxin was an effective inhibitor of progesterone action when microinjected into oocytes, with half-maximal inhibition occurring at an approximate internal concentration of 1 X 10(-7) M. Cholera toxin alone increased cAMP levels by 20%, but upon addition of progesterone, the level increased transiently to 200% of basal, indicating that the inhibition was due to elevated levels of cAMP. The results support a model in which the initiation of meiotic cell division is regulated by cAMP and protein phosphorylation.


In a cohort of individuals with type 2 diabetes using the drug sulfasalazine, HbA1c lowering is associated with haematological changes.
Several small studies indicate the sulphonamide component of the drug sulfasalazine lowers HbA1c. We investigated reduction of HbA1c following incident prescription of sulfasalazine and related aminosalicylates, lacking the sulphonamide group, in an observational cohort. Individuals in the Scottish Care Information Diabetes Collaboration (SCI-Diabetes) with type 2 diabetes and incident prescription for an aminosalicylate drug (sulfasalazine, mesalazine, olsalazine or balsalazide) were identified. Baseline and 6-month HbA1c were required for eligibility, to calculate HbA1c response. To investigate association with haemolysis, change in components of full blood count was assessed. Paired t-tests compared difference in baseline and treatment HbA1c measures and other clinical variables. In all, 113 individuals treated with sulfasalazine and 103 with mesalazine (lacking the sulphonamide group) were eligible, with no eligible individuals treated with olsalazine or balsalazide. Baseline characteristics were similar. Mean (SD) HbA1c reduction at 6 months was -9 ± 16 mmol/mol (-0.9 ± 1.4%) (p < 0.0001) in those taking sulfasalazine with no reduction in those taking mesalazine (2 ± 16 mmol/mol (0.2 ± 1.4%). Sulfasalazine but not mesalazine was associated with a mean (SD) increase in mean cell volume of 3.7 ± 5.6 fl (p < 0.0001) and decrease in red cell count of -0.2 ± 0.4 × 10-12 /L (p < 0.0001). In this observational, population-based study, sulfasalazine initiation was associated with a 6-month reduction in HbA1c . This correlated with haematological changes suggesting haemolytic effects of sulfasalazine. Haemolysis is proposed to contribute to HbA1c lowering through the sulphonamide pharmacophore. This suggests that HbA1c is not a reliable measure of glycaemia in individuals prescribed sulfasalazine.


"Host tissue damage" signal ATP promotes non-directional migration and negatively regulates toll-like receptor signaling in human monocytes.
The activation of Toll-like receptors (TLRs) by lipopolysaccharide or other ligands evokes a proinflammatory immune response, which is not only capable of clearing invading pathogens but can also inflict damage to host tissues. It is therefore important to prevent an overshoot of the TLR-induced response where necessary, and here we show that extracellular ATP is capable of doing this in human monocytes. Using reverse transcription-PCR, we showed that monocytes express P2Y(1), P2Y(2), P2Y(4), P2Y(11), and P2Y(13) receptors, as well as several P2X receptors. To elucidate the function of these receptors, we first studied Ca(2+) signaling in single cells. ATP or UTP induced a biphasic increase in cytosolic Ca(2+), which corresponded to internal Ca(2+) release followed by activation of store-operated Ca(2+) entry. The evoked Ca(2+) signals stimulated Ca(2+)-activated K(+) channels, producing transient membrane hyperpolarization. In addition, ATP promoted cytoskeleton reorganization and cell migration; however, unlike chemoattractants, the migration was non-directional and further analysis showed that ATP did not activate Akt, essential for sensing gradients. When TLR2, TLR4, or TLR2/6 were stimulated with their respective ligands, ATPgammaS profoundly inhibited secretion of proinflammatory cytokines (tumor necrosis factor-alpha and monocyte chemoattractant protein-1) but increased the production of interleukin-10, an anti-inflammatory cytokine. In radioimmune assays, we found that ATP (or ATPgammaS) strongly increased cAMP levels, and, moreover, the TLR-response was inhibited by forskolin, whereas UTP neither increased cAMP nor inhibited the TLR-response. Thus, our data suggest that ATP promotes non-directional migration and, importantly, acts as a "host tissue damage" signal via the G(s) protein-coupled P2Y(11) receptor and increased cAMP to negatively regulate TLR signaling.


Factors affecting the incidence of chronic pain following breast cancer surgery: Preoperative history, anesthetic management, and surgical technique.
Breast cancer is the most frequent cancer in women. Chronic pain following mastectomy remains a significant problem. This study aimed to evaluate risk factors associated with postoperative chronic pain. Medical records were analyzed retrospectively for 147 consecutive patients with breast cancer who underwent mastectomy between July 2016 and February 2018 with one-year follow-up. All patients received a paravertebral block pre-operatively and then general anesthesia for the surgery. The median postoperative pain score was 1.3 on the verbal analgesic score. The average total dose of opioids administered postoperatively was 10.4 mg, expressed in morphine milligram equivalents. The incidence of chronic pain after mastectomy was 16.7% at 6 months and 13.3% at 1 year. The satisfaction rate for anesthesia and surgery was 95.7%. The data suggested that preoperative paravertebral nerve blocks, appropriate adjuvants, and a balanced general anesthetic may contribute to better postoperative pain control and decreased the incidence of chronic pain. A history of preoperative use of opioids and/or a history of alcohol consumption were both associated with significant increased levels of postoperative pain, and this subgroup warrants attention to diminish the risk of developing chronic pain.


Multiple Arterial Thrombosis in Seronegative Antiphospholipid Syndrome: Need for New Diagnostic Criteria?
Antiphospholipid syndrome (APS) is defined as thromboembolic complications and/or pregnancy morbidity in the presence of persistent increased titres of antiphospholipid antibodies. Nevertheless, some patients with clinical signs suggestive of APS are negative for diagnostic antibodies and may be classified as having seronegative-APS (SN-APS). Among the 'non-diagnostic' antibodies, a few studies have suggested that the IgG anti-vimentin/cardiolipin antibodies (AVA/CL) may be associated with risk of thrombosis. The aim of this case report is to encourage the assessment of non-conventional antibodies in APS.Patient and methods: We report the case of a 69-year-old male patient with rapid onset of apparently unexplained multiple exclusively arterial thrombotic events in both coronary and peripheral vascular beds. The patient did not meet the diagnostic criteria for APS but was positive for AVA/CL, which result persisted on further testing at 3 and 6 months. Ongoing research has revealed the existence of non-criteria antibodies which may be relevant for the diagnosis of SN-APS and should be included in the classification criteria for the disease. In patients with unexplained multiple thrombosis without the conventional antibodies of antiphospholipid syndrome, the assessment of non-conventional antibodies should be considered.IgG anti-vimentin/cardiolipin antibodies may be associated with risk of thrombosis.Arterial thrombosis could be the only manifestation of antiphospholipid syndrome.


K(+) starvation inhibits water-stress-induced stomatal closure.
The effect of potassium starvation on stomatal conductance was studied in olive trees and sunflower plants, two major crops with greatly differing botanical characteristics. In both species, K(+) starvation inhibited water-stress-induced stomatal closure. In olive trees, potassium starvation favoured stomatal conductance and transpiration, as well as inhibiting shoot growth, in the three cultivars studied: 'Lechín de Granada', 'Arbequina' and 'Chetoui'. However, 'Lechín de Granada' - generally considered more drought-tolerant than 'Arbequina' and 'Chetoui' - proved less susceptible to potassium starvation. Results for olive trees also suggest genetic variability in olive cultivars in relation to potassium requirements for stem growth and the regulation of water transpiration. The results obtained suggest that inhibition of the stomatal closure mechanism produced by moderate potassium starvation is a widespread plant physiological disorder, and may be the cause of tissue dehydration in many water-stressed crops.


Reliability of Aortic Stenosis Severity Classified by 3-Dimensional Echocardiography in the Prediction of Cardiovascular Events.
The estimation of aortic valve area (AVA) by Doppler echocardiography-derived left ventricular stroke volume (LVSV) remains controversial. We hypothesized that AVA estimated from directly measured LVSV by 3-dimensional echocardiography (3DE) on the continuity equation might be more accurate in classifying aortic stenosis (AS) severity. We retrospectively enrolled 265 patients with moderate-to-severe AS with preserved ejection fraction. Indexed AVA (iAVA) was calculated using LVSV derived by 2D Doppler (iAVADop), Simpson's method (iAVASimp), and 3DE (iAVA3D). During a median follow-up period of 397 days (interquartile range 197 to 706 days), 135 patients experienced the composite end point (cardiac death 9%, aortic valve replacement 24%, and cardiovascular event 27%). Estimated iAVA3D and iAVASimp were significantly smaller than iAVADop and moderately correlated with peak aortic jet velocity. Upper septal hypertrophy was a major cause of discrepancy between iAVADop and iAVA3D methods. Based on the optimal cut-off point of iAVA for predicting peak aortic jet velocity >4.0 m/s, 141 patients (53%) were classified as severe AS and 124 patients (47%) as moderate AS by iAVADop. Indexed AVA3D classified 118 patients (45%) as severe and 147 patients (55%) as moderate AS. Of the 124 patients with moderate AS by iAVADop, 22 patients (18%) were reclassified as severe AS by iAVA3D and showed poor prognosis (hazard ratio 2.7, 95% CI 1.4 to 5.0; p = 0.001). In conclusion, 3DE might be superior in classifying patients with AS compared with Doppler method, particularly in patients with upper septal hypertrophy.


Liver Injury in Patients Hospitalized with Coronavirus Disease 2019 Correlates with Hyperinflammatory Response and Elevated Interleukin-6.
Liver injury is commonly seen in coronavirus disease 2019 (COVID-19); however, the mechanism behind liver injury, particularly in patients with severe and critical COVID-19, remains unclear, and the clinical course is poorly described. We conducted a single-center retrospective cohort study of consecutive patients hospitalized with severe and critical COVID-19 with or without liver injury and who underwent immunologic testing (interleukin [IL]-6, IL-8, tumor necrosis factor alpha [TNF-α], and IL-1β). Liver injury was defined as peak aminotransferases ≥3 times the upper limit of normal (40 U/L) or ≥120 U/L. Patients with liver injury were compared to those who had normal aminotransferases throughout the hospital course. We studied 176 patients: 109 with liver injury and 67 controls. Patients with liver injury were more likely to be men (71.6% vs. 37.3%, P < 0.001). Peak inflammatory markers and IL-6 were higher in the liver injury group: C-reactive protein (CRP), 247 vs. 168 mg/L, P < 0.001; lactate dehydrogenase (LDH), 706 vs. 421 U/L; ferritin, 2,973 vs. 751 ng/mL, P < 0.001; IL-6, 121.0 vs. 71.8 pg/mL, P < 0.001. There was no difference in the levels of IL-8, TNF-α, and IL-1β. The liver injury group had a longer length of stay in the hospital and more severe COVID-19 despite having less diabetes and chronic kidney disease. Conclusion: An exaggerated hyperinflammatory response (cytokine storm) characterized by significantly elevated CRP, LDH, ferritin, and IL-6 levels and increasing severity of COVID-19 appears to be associated with the occurrence of liver injury in patients with severe/critical COVID-19.


Comprehension of the copula: preschoolers (and sometimes adults) ignore subject-verb agreement during sentence processing.
Subject-verb (SV) agreement helps listeners interpret the number condition of ambiguous nouns (The sheep is/are fat), yet it remains unclear whether young children use agreement to comprehend newly encountered nouns. Preschoolers and adults completed a forced choice task where sentences contained singular vs. plural copulas (Where is/are the [novel noun(s)]?). Novel nouns were either morphologically unambiguous (tup/tups) or ambiguous (/geks/ = singular: gex / plural: gecks). Preschoolers (and some adults) ignored the singular copula, interpreting /ks/-final words as plural, raising questions about the role of SV agreement in learners' sentence comprehension and the status of is in Australian English.


Adenylate cyclase activity of cultural strains of Neisseria gonorrhoeae and Staphylococcus aureus.
Adenylate cyclase of gonococci and staphylococci from patients with gonorrhoeae and pyoderma was studied ultracytochemically. Adenylate cyclase activity was mainly revealed on the inner side of the plasma membrane and septa of dividing cells. The enzymic activity is markedly higher in penicillinase producing strains and dividing bacteria. These data may be useful for the diagnostics and treatments of the infections concerned.


Severity and natural history of acute pancreatitis in diabetic patients.
There is limited data on the prevalence of coexistent diabetes in acute pancreatitis and subsequent natural history in these patients. Using Pennsylvania Health Care Cost Containment Council data set, we identified 7399 unique White and Black Allegheny County, PA residents with first hospitalization for acute pancreatitis from 1996 to 2005. We evaluated the prevalence of coexistent diabetes, demographic and etiologic profile, severity of index hospitalization, and risk of readmission for acute or chronic pancreatitis during follow up (median 39 months) in this cohort. The prevalence of coexisting diabetes was 18% (1349/7399). When compared with non-diabetics, diabetics were significantly more likely to be older (63 vs. 56 yrs), male (OR 1.4, 95% CI 1.2-1.6), black (OR 2.4, 95% CI 2.1-2.7) and have non-alcoholic etiologies (biliary, OR 1.5, 95% CI 1.2-1.9; idiopathic, OR 2.0, 95% CI 1.7-2.5; metabolic, OR 5.2, 95% CI 4.0-6.7). While diabetic patients had a significantly longer length of stay (median 5 vs. 4 days, p < 0.05), their severity of acute pancreatitis (multivariable OR 1.18, 95% CI 0.94-1.48) or in-hospital mortality (1.9% each, p = 0.98) did not differ than non-diabetics. The overall risk of pancreatitis-related readmissions in diabetics was similar (33% each, p = 0.99), but their risk of subsequent admission for chronic pancreatitis (multivariable HR 0.65, 95% CI 0.44-0.97) was lower than non-diabetics. About 20% patients with acute pancreatitis have coexistent diabetes. Demographic and etiologic profile of diabetic patients with acute pancreatitis differs from non-diabetics. Diabetic status has limited effect on the severity of and natural history following acute pancreatitis.


Body ideals in women after viewing images of typical and healthy weight models.
Viewing thin models, pervasive in popular culture, is correlated with body dissatisfaction and anxiety in women. Whether or not the same is true when viewing healthy weight models is unknown. In this study we tested whether viewing healthy weight models increases the ideal female body size. Body image, anxiety, happiness and depression were measured in 44 female participants following viewing of images of thin or healthy weight models (within-subject separated by two weeks). We found that after viewing images of healthy weight models, women's body ideals (as measured by a participant-adjusted virtual model) were significantly larger than when the same women viewed images of very thin models. This effect was greatest in those women with the highest levels of baseline anxiety (as measured by the Hospital Anxiety and Depression Scale). These results suggest that viewing healthy weight models results in more healthy body ideals than those typically promoted through media.


Dynamic simulation of pollutant effects on the threonine pathway in Escherichia coli.
The enzymatic activities of threonine pathway in Escherichia coli are sensitive to pollutants such as cadmium, copper and mercury, which, even at low concentration, can substantially decrease or even block the pathway at several steps. Our aim was to investigate the complex effects on a metabolic pathway of such general enzyme inhibitors with several sites of action, using a previously developed computer simulation of the pathway. For this purpose, the inhibition parameters were experimentally determined and incorporated in the model. The calculation of the flux control coefficient distribution between the five steps of the threonine pathway showed that control remains shared between the three first steps under most inhibition conditions. Response coefficient analysis shows that the inhibition of aspartate semialdehyde dehydrogenase is quantitatively dominant in most circumstances.


Risks associated with communication delays in infants from underserved South African communities.
For optimal development young children need warm, responsive, enriched and communicative environments for learning social, language, and other skills. Infants and toddlers exposed to psychosocial risk lack enriched environments and may present with communication delays. To investigate the relationship between psychosocial risks and communication delays in infants from underserved communities in South Africa. Primary healthcare facilities in Tshwane district, South Africa. A parent interview and Rossetti Infant Toddler Language Scales were used to collect data from caregivers of 201 infants aged 6–12 months, selected through convenience sampling. Associations between communication delays and risks were determined (Chi-square and Fisher's exact tests). A log-linear model analysis was used to model the simultaneous effect of significant risks on the probability of having communication delays. Communication delays were present in 13% of infants. Infants with two or more siblings, born from mothers aged 18–29 years who own their house, had a 39% chance of presenting with communication delays. Developmental screening and early intervention is important in primary healthcare contexts in South Africa, as a clear relationship has been established between three risk factors and communication delays in infants.


Premature infants with gestational age less than 25 weeks require increased ophthalmology resources for retinopathy of prematurity.
It is unclear how increasing survival of low gestational age (GA) infants affects ophthalmologic screening and treatment rates for retinopathy of prematurity (ROP). This study compared the examination and treatment rates of infants born at GA of <25 weeks and those born at GA of at least 25 weeks. This was a retrospective study of patients who met institutional ROP screening criteria and were admitted to two neonatal intensive care units (NICUs) from January 2017 to June 2020. Variables analyzed were GA, birth weight, number of ophthalmology examinations, worst stage of ROP, presence of type 1 ROP, and comorbidities associated with ROP. The χ2, Fisher exact, and two-tailed t tests, as well as univariate and multivariable logistic regression, were used for statistical analysis. Compared to the GA≥25 group, the GA<25 group had a higher number of total exams (10 vs 4.3 [P < 0.001]), higher average worst stage of ROP (1.4 vs 0.3 [P < 0.001]) and higher rate of type 1 ROP (21% vs 1.4% [P < 0.001]), as well as higher mortality (37% vs 8.11% [P < 0. 001]). Multivariable logistic regression analysis controlling for GA, sepsis, and number of transfusions revealed that only GA was significantly associated with developing type 1 ROP. Infants with GA <25 weeks had more severe ROP and required significantly more ophthalmologic examinations than GA ≥25. It is important for ROP services to plan for this increased screening load, especially if the number of such lower-weight infants in their NICUs increases.


A broader role for advance medical planning.
Advance planning for future illness should be broadened from medical care in the event of incompetence to all medical care for the elderly. To plan effectively, patients need an assessment of their overall medical condition: whether they are robust, frail, demented, or dying. They need to understand the kinds of complications often engendered by aggressive treatment, given their underlying status. Given information about their circumstances and their capacity to withstand medical interventions, patients, together with their physicians, need to formulate broad goals for medical care. There are significant barriers to implementing this scheme, but pressure from patients, structural changes in the practice of medicine that create incentives for planning, and educational strategies, including videotaped interviews and role-playing exercises, can facilitate this form of preventive medicine.


Perinatal exposure to low doses of tributyltin chloride advances puberty and affects patterns of estrous cyclicity in female mice.
Tributyltin (TBT), a proven endocrine-disrupting chemical, is well known to induce imposex in female gastropods. Herein we demonstrate the effects of low doses of tributyltin chloride (TBTCl) on the female offspring of KM mice. Pregnant mice were administered by gavage with 0, 1, 10, or 100 μg TBTCl/kg body weight/day from day 6 of pregnancy through the period of lactation. TBTCl dramatically advanced the age of onset of vaginal opening (VO) and first vaginal estrus, and reduced body weights at VO and first estrus. Furthermore, perinatal treatment with TBTCl significantly reduced the number of days between VO and first estrus. In addition, female offspring from dams exposed to 10 and 100 μg kg(-1) TBTCl exhibited altered patterns of estrous cyclicity in adulthood. In conclusion, perinatal exposure to low doses TBTCl result in early puberty and impaired estrous cyclicity in female mice, which suggest that TBTCl might act as an estrogen agonist or/and a disruptor on hypothalamic-pituitary function in the present study.


Utilization of the emergency department as a routine source of care among children with asthma.
Objective: To describe characteristics of children with persistent asthma in the ED who receive most of their healthcare in emergency settings; and determine whether recent asthma experiences or historic patterns of care are associated with identifying the ED as a typical location for care.Methods: We conducted a sub-analysis of baseline data from Telemedicine Enhanced Asthma Management through the Emergency Department (TEAM-ED), an RCT of children (3-12 years) presenting to the ED with persistent asthma (2016-2020). Caregivers identified reasons for seeking emergency care, including if their child received most overall healthcare in the ED ('ED Care'; primary outcome) or not ('Other Care'). Independent variables included demographics, recent symptoms and quality of life (QOL), and historic preventive care and healthcare use. We compared responses between ED Care and Other Care groups using bivariate and multivariate analyses.Results: We analyzed data for 355 children (31% ED Care, 69% Other Care). Compared with Other Care, ED Care respondents were more likely to identify the ED as the closest source of healthcare; report fewer symptom nights but a poorer quality of life; and describe the ED as a usual place for sick care, despite most having a PCP.Conclusions: Many children with asthma use the ED as a typical source of healthcare, and are distinguished by need for proximity, poorer caregiver QOL, and historic patterns of care-seeking. Efforts to improve timely access to outpatient care and reinforce the role of PCP-directed asthma management, such as through telemedicine, may reduce preventable morbidity including ED visits.


Sc3+-catalyzed aldol-type additions of N-benzoylcyclopropanecarboxamides via iodide-mediated ring-opening: stereoselective synthesis of gamma-lactams.
A new catalytic aldol-type addition of cyclopropanecarboximides to aldehydes via iodide-mediated ring-opening is presented. The reaction was found to be catalyzed at 0 degrees C using either a Sc(OTf)3/NaI system or ScI3. Stereoselective formation of alpha,alpha-disubstituted enolates occurred in situ. gamma-Lactams bearing alpha-carbonyl quaternary stereocenters were obtained in 97-57% yield and dr = 90:10-80:20 after ring closure.


Psychological and psychiatric aspects of pain.
The relationship of pain to personality and the significance have been discussed. Pain may first be evidence of tissue damage, secondly it may be used as a means of communicating emotional distress to others and thirdly, it may be a means of manipulating others, expressing hostility or relieving guilt. The evidence is clear that an understanding of mental life and its relation to pain is important if this symptoms is to be dealt with effectively, irrespective of whether it is physical or psychogenic in origin.


Twinkle helicase (PEO1) gene mutation causes mitochondrial DNA depletion.
Mitochondrial DNA (mtDNA) depletion syndrome (MDS) is a clinically and genetically heterogeneous group of autosomal recessive diseases characterized by a reduction in mtDNA copy number. Several nuclear genes have been shown to account for these severe oxidative phosphorylation disorders, but the disease-causing mutations remain largely unknown. By virtue of homozygosity mapping, we tested candidate genes involved in mtDNA maintenance in patients born to consanguineous parents. We found homozygosity for microsatellite markers flanking the PEO1 gene, encoding the mitochondrial Twinkle helicase, in two sibs presenting a hepatocerebral form of MDS. Sequencing the PEO1 gene showed a homozygous mutation at a conserved position of the protein in the two patients (T457I). The modeling of the Twinkle protein showed that T457 is located in the interface between two monomers of the hexameric enzyme. Finally, using purified recombinant protein, we demonstrated that the T457I mutant Twinkle has a defective helicase activity. Although dominant Twinkle mutations have been previously reported in patients with autosomal dominant progressive external ophthalmoplegia and multiple mtDNA deletions, we report here the first recessive Twinkle mutation in patients with hepatocerebral form of MDS. Identifying other Twinkle mutations in MDS and/or autosomal dominant progressive external ophthalmoplegia and studying their impact on the isolated proteins should help in understanding why some mutations are recessive and others are dominant.


Starburst amacrine cells amplify optogenetic visual restoration through gap junctions.
Ectopic induction of optogenetic actuators, such as channelrhodopsin, is a promising approach to restoring vision in the degenerating retina. However, the cell type-specific response of ectopic photoreception has not been well understood. There are limits to obtaining efficient gene expression in a specifically targeted cell population by a transgenic approach. In the present study, we established a murine model with high efficiency of gene induction to retinal ganglion cells (RGCs) and amacrine cells using an improved tetracycline transactivator-operator bipartite system (KENGE-tet system). To investigate the cell type-specific visual restorative effect, we expressed the channelrhodopsin gene into RGCs and amacrine cells using the KENGE-tet system. As a result, enhancement in the visual restorative effect was observed to RGCs and starburst amacrine cells. In conclusion, a photoresponse from amacrine cells may enhance the maintained response of RGCs and further increase or improve the visual restorative effect.


In vivo neurochemical analysis, by push-pull perfusion, of the mesocortical dopaminergic system of the rat during self-stimulation.
The region immediately adjacent to a self-stimulation site in the medial prefrontal cortex of the unanesthetized rat was prelabeled with 0.5 mu Ci 14C-dopamine (DA) injected through an indwelling guide cannula. Then successive 5 min push-pull perfusions of the site with an artificial CSF were carried out at a rate of 25 microliter/min so that a washout curve of declining radioactivity was generated under control conditions. When square wave 100 Hz pulses were delivered to the contiguous self-stimulation site, the release of 14C-DA was enhanced either during the actual interval of electrical stimulation or in the perfusion sample collected immediately thereafter. In parallel experiments, however, self-stimulation by the rat of its ventral tegmental area failed to alter the kinetics of 14C-DA release from the cortex when homologous loci were perfused. Analyses by thin-layer chromatography of the perfusates for their content of catechol metabolities revealed that the homovanillic acid fraction declined during stimulation, whereas the level of DOPAC remained relatively elevated. Evidence was also obtained for the new synthesis and subsequent release of norepinephrine during the stimulation of the cortex of the rat. These results suggest that endogenous dopamine, because of the notable alterations in its release and metabolism, plays an important synaptic role in the mediation of self-stimulation behavior at the level of the cerebral cortex.


New insight into protein glycosylation in the development of Alzheimer's disease.
Alzheimer's disease (AD) is a chronic neurodegenerative disease that seriously endangers the physical and mental health of patients, however, there are still no effective drugs or methods to cure this disease up to now. Protein glycosylation is the most common modifications of the translated proteins in eukaryotic cells. Recently many researches disclosed that aberrant glycosylation happens in some important AD-related proteins, such as APP, Tau, Reelin and CRMP-2, etc, suggesting a close link between abnormal protein glycosylation and AD. Because of its complexity and diversity, glycosylation is thus considered a completely new entry point for understanding the precise cause of AD. This review comprehensively summarized the currently discovered changes in protein glycosylation patterns in AD, and especially introduced the latest progress on the mechanism of protein glycosylation affecting the progression of AD and the potential application of protein glycosylation in AD detection and treatment, thereby providing a wide range of opportunities for uncovering the pathogenesis of AD and promoting the translation of glycosylation research into future clinical applications.


Interaction of the hemochromatosis gene product HFE with transferrin receptor modulates cellular iron metabolism.
Mutations of a novel MHC class I-like protein, termed HFE, have been found in the vast majority of patients with the iron overload disease heredity hemochromatosis. Identification of HFE is likely to shed light on one of the major enigmas of mammalian iron homeostasis: How is intestinal iron absorption regulated?


[The effect of the osmotic pressure of the culture medium on cells of Francisella tularensis vaccinal strain 15].
The dependence of the growth rate of F. tularensis on the osmotic properties of the medium can be presented as a curve with the maximum in the area of 500-600 mOsm. Under these circumstances the intracellular osmotic pressure exceeds the extracellular one by 50-100 mOsm. With the rise of the osmotic pressure in the medium the increase of the concentration of K+ in the cells occurs. The energy-dependent accumulation of K+ in the cells at rest is activated by the rise of the osmotic pressure in the medium. F. tularensis are probably capable of osmoregulation, ensured by the energy-dependent osmosensitive K(+)-transporting system.


Overview of regulatory strategies and molecular elements in metabolic engineering of bacteria.
From a viewpoint of biotechnology, metabolic engineering mainly aims to change the natural status of a pathway in a microorganism towards the overproduction of certain bioproducts. The biochemical nature of a pathway implies us that changed pathway is often the collective results of altered behavior of the metabolic enzymes encoded by corresponding genes. By finely modulating the expression of these genes or the properties of the enzyme, we can gain efficient control on the pathway. In this article, we reviewed the typical methods that have been applied to regulate the expression of genes in metabolic engineering. These methods are grouped according to the operation targets in a typical gene. The transcription of a gene is controlled by an indispensable promoter. By utilizing promoters with different strengths, expected levels of expression can be easily achieved, and screening a promoter library may find suitable mutant promoters that can provide tunable expression of a gene. Auto-responsive promoter (quorum sensing (QS)-based or oxygen-inducible) simplifies the induction process by driving the expression of a gene in an automated manner. Light responsive promoter enables reversible and noninvasive control on gene activity, providing a promising method in controlling gene expression with time and space resolution in metabolic engineering involving complicated genetic circuits. Through directed evolution and/or rational design, the encoding sequences of a gene can be altered, leading to the possibly most profound changes in properties of a metabolic enzyme. Introducing an engineered riboswitch in mRNA can make it a regulatory molecule at the same time; ribosomal binding site is commonly engineered to be more attractive for a ribosome through design. Terminator of a gene will affect the stability of an mRNA, and intergenic region will influence the expression of many related genes. Improving the performance of these elements are generally the main activities in metabolic engineering.


Studies with a spontaneous murine tumour--II. Transformation of non-inhibitory lymph node cells (LNC) of tumour bearers into tumour inhibitor cells in vitro.
Syngeneic BALB/c mice repeatedly pre-exposed to mitomycin-c-treated cells of a 'spontaneous' tumour SP/N-1 failed to develop resistance against the same tumour when tested by cell transfer assays. No difference could be observed between tumour-bearer, tumour-pretreated or normal donor LNC, or spleen cells by this assay system. When tumour-bearer LNC were cultured in vitro for 45 hr in the absence of tumour cells, these LNC generated a relatively strong ability to suppress tumour outgrowth in vivo following cell transfers. The tumour suppression appears to be due to tumour cell destruction since observation over four months or more in some experiments failed to reveal the presence of tumours at the site of inoculation. These findings clearly demonstrate that naturally arising tumours so often considered as devoid of, or weak in, tumour-associated transplantation antigens (TATA) may not necessarily be so. Despite the fact that the syngeneic host may be capable of recognizing TATA as foreign, the immune response initiated may not progress to completion in at least a proportion of these tumour-host systems. The defect appears to reside in the failure of differentiation of the pre-effector cells into the mature effector form. We interpret this as being due to a strong activation of certain inhibitors of essential lymphokine mediators such as interleukin 2, needed for differentiation and expansion of effector cell clones. It remains to be seen whether the mechanism of non-responsiveness or weakness of the response against many other members of this category of tumours--the 'spontaneous' or natural tumours--can be explained on the same basis as that of the SP/N-1 system studied here.


Behavioral and physiological consequences of suckling in rat and human newborns.
Suckling, in addition to yielding milk, water and calories, exerts profound behavioral effects on newborn rats and humans. In particular, suckling induces feelings of calm, reduces heart rate and metabolic rate, causes infants to bring their hands to their mouths and elevates the pain threshold. These changes are mediated by opioid and non-opioid systems, each having its own separate behavioral and neurological characteristics. The implications of suckling-induced changes for long-term motivational and cognitive change are discussed.


The effects of movements on caudate sensory responses.
The aim of this experiment was to assess the effects of movement on somatosensory processing in the caudate nucleus. Units were recorded extracellularly in the caudate of awake partially restrained cats. Unit responses were evoked by either natural stimuli (brushing and skin indentation) or transcutaneous electrical stimulation. Response patterns evoked by stimuli presented when the cat was at rest were compared to responses evoked by similar stimuli presented when the cat was either making buccolingual movements or turning its head. In the majority of cases, movement suppressed caudate somatosensory responses. However, impressive augmentation of responses was seen in some cells with either type of movement.


Intratracheal budesonide mixed with surfactant to increase survival free of bronchopulmonary dysplasia in extremely preterm infants: study protocol for the international, multicenter, randomized PLUSS trial.
Bronchopulmonary dysplasia (BPD), an inflammatory-mediated chronic lung disease, is common in extremely preterm infants born before 28 weeks' gestation and is associated with an increased risk of adverse neurodevelopmental and respiratory outcomes in childhood. Effective and safe prophylactic therapies for BPD are urgently required. Systemic corticosteroids reduce rates of BPD in the short-term but are associated with poorer neurodevelopmental outcomes if given to ventilated infants in the first week after birth. Intratracheal administration of corticosteroid admixed with exogenous surfactant could overcome these concerns by minimizing systemic sequelae. Several small, randomized trials have found intratracheal budesonide in a surfactant vehicle to be a promising therapy to increase survival free of BPD. An international, multicenter, double-blinded, randomized trial of intratracheal budesonide (a corticosteroid) mixed with surfactant for extremely preterm infants to increase survival free of BPD at 36 weeks' postmenstrual age (PMA; primary outcome). Extremely preterm infants aged < 48 h after birth are eligible if: (1) they are mechanically ventilated, or (2) they are receiving non-invasive respiratory support and there is a clinical decision to treat with surfactant. The intervention is budesonide (0.25 mg/kg) mixed with poractant alfa (200 mg/kg first intervention, 100 mg/kg if second intervention), administered intratracheally via an endotracheal tube or thin catheter. The comparator is poractant alfa alone (at the same doses). Secondary outcomes include the components of the primary outcome (death, BPD prior to or at 36 weeks' PMA), potential systemic side effects of corticosteroids, cost-effectiveness, early childhood health until 2 years of age, and neurodevelopmental outcomes at 2 years of age (corrected for prematurity). Combining budesonide with surfactant for intratracheal administration is a simple intervention that may reduce BPD in extremely preterm infants and translate into health benefits in later childhood. The PLUSS trial is powered for the primary outcome and will address gaps in the evidence due to its pragmatic and inclusive design, targeting all extremely preterm infants regardless of their initial mode of respiratory support. Should intratracheal budesonide mixed with surfactant increase survival free of BPD, without severe adverse effects, this readily available intervention could be introduced immediately into clinical practice. Australian New Zealand Clinical Trials Registry ( https://www.anzctr.org.au ), ACTRN12617000322336. First registered on 28th February 2017.


Scanning electron microscopy of the internal wall topography of enteral feeding tubes.
A range of commercially available naso-gastric and naso-enteric tubes was examined by scanning electron microscopy (S.E.M.). Surface irregularities in which micro-organisms could become trapped were observed on the internal walls of all the samples. Fine bore tubes which had been perfused for 8 h with a milk based feed experimentally contaminated with Staphylococcus aureus showed patches of residual feed and S. aureus cells on their interior surfaces. The potential hazards of entrapment and/or attachment of micro-organisms to the interior walls of these tubes are discussed.


Binocular implantation of the Tecnis Z9000 or AcrySof MA60AC intraocular lens in routine cataract surgery: prospective randomized controlled trial comparing VF-14 scores.
To compare vision-related quality of life after binocular implantation of the Tecnis Z9000 intraocular lens (IOL) (Advanced Medical Optics) or the AcrySof MA60AC IOL (Alcon). Moorfields Eye Hospital, London, United Kingdom. The study was a double-masked prospective randomized controlled trial. The main outcome measure was the percentage of patients achieving a Visual Function-14 (VF-14) score of 90 or higher. Additional outcome measures were dysphotopsia symptom scores, a 5-point subjective grading of overall satisfaction, refraction, photopic and mesopic best corrected visual acuities, uncorrected visual acuity, Pelli-Robson contrast sensitivity, aberrometry, infrared pupillometry, and evaluation of posterior capsule opacification scores. Seventy-nine patients who had bilateral implantation of the Z9000 IOL and 100 patients who had bilateral implantation of the MA60AC IOL were assessed a mean of 7.9 months +/- 2.9 (SD) (range 4 to 16 months) after second-eye surgery. Spherical aberration (mean: Z9000, 0.00 microm +/- 0.06 [SD]; MA60AC, +0.14 +/- 0.09 microm) and root mean square scores for higher-order aberrations combined (mean: Z9000, 0.26 +/- 0.11 microm; MA60AC, 0.32 +/- 0.14 microm) were significantly lower in patients with the Z9000 IOL (P<.001 and P = .007, respectively). The VF-14 scores were 90 or higher in 96.2% of Z9000 patients and 90.0% of MA60AC patients (P = .15). It is unlikely that this weak trend in favor of the Z9000 IOL was clinically significant. The VF-14 scores were heavily skewed toward a perfect outcome and similarly distributed in both groups (median 100; interquartile range 98 to 100). Results for all other outcomes, including measures of acuity, were similarly distributed in Z9000 patients and MA60AC patients. The Z9000 IOL reduced spherical aberration compared with the MA60AC IOL. No improvement in subjective visual function using the VF-14 questionnaire was detected.


Cyclosporin A is a differential inhibitor of eukaryotic RNA polymerases.
In order to investigate the molecular mechanism of biological action of the drug cyclosporin A (CsA), we have studied its effect on eukaryotic RNA polymerase activities in two systems: (a) in vitro transcription; (b) injection into frog oocytes. The electron microscopic transcription R-loop method [1] has been used to assay for specific transcription of an immunoglobulin gene clone in HeLa cell lysates. In this system, CsA inhibits efficiently the transcription of immunoglobulin genes. Dilution experiments show that RNA polymerase II activity is inhibited by nanomolar concentrations (i.e. amounts comparable to the ones needed for the inhibition of lymphocyte stimulation), whereas polymerase III is inhibited at micromolar concentrations. In vitro transcription with E. coli RNA polymerase is not inhibited. Parallel experiments with the mushroom toxin alpha-amanitin have shown that the eukaryotic RNA polymerase II displays similar sensitivity towards the two cyclic peptides CsA and alpha-amanitin in vitro. When CsA is injected into frog oocytes, together with a cloned Xenopus laevis U1 small nuclear RNA gene, transcription of this gene by endogenous RNA polymerase II is inhibited by 60%. RNA polymerase III and RNA polymerase I transcription is not inhibited in the oocyte. Various possibilities for the selective action of CsA on stimulated lymphocytes are discussed.


Leveraging intrinsic flexibility to engineer enhanced enzyme catalytic activity.
Dynamic motions of enzymes occurring on a broad range of timescales play a pivotal role in all steps of the reaction pathway, including substrate binding, catalysis, and product release. However, it is unknown whether structural information related to conformational flexibility can be exploited for the directed evolution of enzymes with higher catalytic activity. Here, we show that mutagenesis of residues exclusively located at flexible regions distal to the active site of Homo sapiens kynureninase (HsKYNase) resulted in the isolation of a variant (BF-HsKYNase) in which the rate of the chemical step toward kynurenine was increased by 45-fold. Mechanistic pre–steady-state kinetic analysis of the wild type and the evolved enzyme shed light on the underlying effects of distal mutations (>10 Å from the active site) on the rate-limiting step of the catalytic cycle. Hydrogen-deuterium exchange coupled to mass spectrometry and molecular dynamics simulations revealed that the amino acid substitutions in BF-HsKYNase allosterically affect the flexibility of the pyridoxal-5′-phosphate (PLP) binding pocket, thereby impacting the rate of chemistry, presumably by altering the conformational ensemble and sampling states more favorable to the catalyzed reaction.


The Effect of Aβ₁₋₄₂ Oligomers on APP Processing and Aβ₁₋₄₀ Generation in Cultured U-373 Astrocytes.
Amyloid-β (Aβ) peptides are a family of proteins that are considered to be a principal aspect of Alzheimer's disease (AD), the most common cause of senile dementia affecting elderly individuals. These peptides result from the proteolytic processing of amyloid precursor protein (APP) by sequential cleavage mediated via β- and x03B3;-secretases. Evidence suggests that an overproduction and/or a lack of degradation may increase brain Aβ levels which, in turn, contribute to neuronal loss and development of AD. In this study, we seek to determine what effect Aβ has on APP processing in cultured astrocytes. Using the human astrocytoma cell line U-373, we investigated the effects induced by oligomeric Aβ1-42 treatment on the cellular levels/expression of APP and its products, C-terminal fragments αCTF and βCTF, and Aβ1-40. In conjunction with these experiments, we examined the relative levels and activity of β- and x03B3;-secretases in Aβ-treated astrocytes. We report here that Aβ1-42 treatment of astrocytes increased the expression of APP and its cleaved products including Aβ1-40 in a time-dependent manner. These results suggest that activated astrocytes can contribute to the development of AD by enhancing levels and processing of APP leading to an increased production/secretion of Aβ-related peptides.


Inguinofemoral sentinel lymph node biopsy by scar injection in vulvar cancer: an assessment of feasibility and long-term outcomes.
Performing inguinofemoral sentinel lymph node biopsy for vulvar cancer following a previous vulvar excision, often referred to as 'scar injection', is debated. To assess the feasibility of sentinel lymph node biopsy following scar injection and the long-term outcomes in patients undergoing this procedure. We conducted a retrospective observational cohort study of patients with vulvar cancer. We assessed detection rates and outcomes in patients who underwent sentinel lymph node biopsy by scar injection and compared them with patients who had injection around a visible tumor and with patients who had an inguinofemoral lymphadenectomy following previous vulvar excision. Sentinel node detection rates are described per patient and per groin and are compared using Χ2 analysis. Cox regression analysis was used to assess the association of recurrence and survival with surgical technique and recognized pathological variables. Data were analyzed for 173 groins in 97 patients. At least one sentinel lymph node was detected in 162 (94%) groins examined, and detection rate did not differ whether the groin was assessed following tumor injection (n=122, 94%) or scar injection (n=40, 93%; p=0.85). Patients in the scar-injection group had less frequent lymph node metastases (p<0.02), smaller tumors (p<0.001), and more superficial invasion (p<0.02). Median follow-up was 34.7 months (range 0-108). Scar injection was not independently associated with recurrence or death on multivariable analysis, and depth of invasion was the only independent predictor of disease recurrence (hazards ratio (HR)=1.14, p=0.03). Recurrence and survival were also comparable for patients who had a sentinel lymph node biopsy or inguinofemoral lymphadenectomy following previous vulvar excision (log rank p=0.30; p=0.67). Sentinel lymph node biopsy by scar injection is feasible and demonstrates similar long-term outcomes in patients having scar or tumor injections, and in patients following previous tumor excision undergoing sentinel lymph node biopsy or lymphadenectomy.


Doppler tissue evaluation of intra-atrial and interatrial electromechanical delay and comparison with P-wave dispersion in patients with mitral stenosis.
The aim of our study was to: (1) measure atrial electromechanical delay in patients with mitral stenosis (MS) and in a control group; (2) find the echocardiographic parameters that affect atrial electromechanical delay; and (3) examine the correlation between atrial electromechanical delay and P-wave dispersion (PWD). A total of 25 patients with pure MS (age 43 +/- 10 years; 18 women, 7 men) and 16 control subjects (age 41 +/- 8 years; 9 women, 7 men) were studied. Interatrial and intra-atrial electromechanical delay was measured with Doppler tissue echocardiography. From the 12-lead electrocardiograms, PWD was calculated. Interatrial electromechanical delay was 71.2 +/- 33 in the MS group and 40.5 +/- 21.0 in the control group (P = .01). In the MS group, PWD was 50 +/- 7 and in the control group it was 29 +/- 5 (P = .03). A positive correlation was detected between interatrial electromechanical delay and PWD (r = 0.6, P = .03). This study shows that interatrial electromechanical delay gets longer in MS and is correlated with PWD. Atrial electromechanical delay is related with left atrial size but not with severity of MS.


Diagnosis and Treatment of Anxiety in the Aging Woman.
The peri- and postmenopausal periods represent a window of vulnerability for emergence of anxiety symptoms and disorders in the life cycle of adult women. Compared to depression, anxiety symptoms and disorders remain largely unexplored during this phase of a woman's life, despite the significant impact on quality of life if not diagnosed and treated. Here, we review the literature to present our current understanding of the epidemiology, causal factors, diagnosis, and treatment of anxiety in the aging woman.


Leprosy and tuberculosis control scenario of the national program for the improvement of access and quality of primary care in Brazil.
In Brazil, despite advances in public health policies aimed at eliminating and controlling infectious and parasitic diseases, the incidence of neglected diseases is still high. The epidemiological scenario in Brazil of diseases such as tuberculosis and leprosy evidences a public policy agenda that has not been resolute in terms of control, nor in terms of elimination. To analyze the actions of diagnosis and treatment of leprosy and tuberculosis in the context of primary health care. In this ecological study, data from the third cycle of the Program for the Improvement of Access and Quality of Primary Care were extracted from electronic address of the Primary Health Care Secretariat of Brazil in the area of Actions, Programs and Strategies. A total of 37,350 primary health care teams were that answered the questionnaire were eligible, with variables extracted from leprosy and tuberculosis control actions. The municipalities were grouped according to the characteristic of the Brazilian municipality. The partition chi-square and the Residuals Test were used to assess whether there was a difference in the proportion of tuberculosis and leprosy actions between types of municipalities. Statistics were carried out using Minitab 20 and Bioestat 5.3. Regarding the leprosy treatment location, there is a higher proportion of people referred to be treated at the reference in adjacent rural (p = 0.0097) and urban (p < 0.0001) municipalities; monitoring of people with leprosy referred to the service network (p. = 0.0057) in remote rural areas. Lower proportion of teams requesting bacilloscopy in remote rural areas (p = 0.0019). Rural areas have a higher proportion of teams that diagnose new cases (p = 0.0004). Regarding the actions of diagnosis and treatment of tuberculosis. There is a higher proportion of teams that carry out consultations at the unit itself in rural areas when compared to adjacent intermediaries (p = 0.0099) and urban (p < 0.0001); who requested sputum smear microscopy in adjacent intermediaries (p = 0.0021); X-ray in adjacent intermediaries (p < 0.0001) and urban (p < 0.0001); collection of the first sputum sample in urban (p < 0.0001) and adjacent rural areas (p < 0.0001); directly observed treatment (p < 0.0001) in adjacent rural municipalities. There are inequalities in the diagnosis and treatment of leprosy and tuberculosis among the types of municipalities.


Modeling above-canopy CO2 flux and evapotranspiration in wheat.
Simulations of above-canopy water vapor and CO2 fluxes were calculated by the USGF linked model of canopy gas exchange and subsurface processes for the 1996-1997 winter wheat season at the AmeriFlux Wheat Site, Oklahoma. Soil surface CO2 flux plus canopy gas exchange and transpiration plus soil evaporation modeled the CO2 and water vapor fluxes, respectively. Parameter values for net photosynthesis, respiration and transpiration were obtained from published sources, generated from Wheat Site data, or estimated by minimizing standard deviation between model and data. The mean measured downward flux of CO2 during rapid growth and maturity of the crop was -0.45 mg m(-2) s(-1) compared to simulated flux of -0.47. Simulated downward CO2 flux exceeded measured values during rapid growth of the crop but underestimated the flux during maturity. For the entire 285-day period, the mean measured upward CO2 flux at night was 0.06 and simulated flux was 0.05.


Cooperative neural processes involved in stereoscopic acuity.
Results of psychophysical experiments are reported showing that synchrony, appropriate relative placement, and absence of standing disparity are important conditions to be met by members of a target configuration if they are to participate in the cooperative neural processes leading to the best disparity discrimination. Consecutive binocular presentation of the members of a stereo target decreases stereoacuity by a factor of about 10, and a step disparity displacement of a single line target needs to be larger still to be detected as a depth stimulus. A standing disaprity of even one minute of arc at least doubles the disaprity disxrimination threshold. It is postulated that a differencing mechanism operates on the depth signal of individual features; the temporal and spatial optima of target presentation for stereoscopic acuity outline the character of the concerned operations.


A spectrophotometric method for the determination of hydroperoxides in liposomes.
A modification of the Asakawa-Matsushita iodometric assay method for the determination of the content of lipid hydroperoxides was developed which permits the simultaneous processing of many samples of high lipid content. The method has the advantages of simplicity as well as good reproducibility, so it is not necessary to process standards with each determination. Our technique exceeds the sensitivity attained with other spectrophotometric determinations reported in the literature. The method requires the total elimination of water from the samples, and this was accomplished using an azeotropic mixture of ethanol:water of 96:4. The results obtained with liposomes indicate that the method is applicable to biological material limited to small volume samples, ranging 5-50 microliters. We want to emphasize that this method permits the study of the peroxidation process as function of time.


What can rodent models tell us about apathy and associated neuropsychiatric symptoms in Parkinson's disease?
In addition to classical motor symptoms, Parkinson's disease (PD) patients display incapacitating neuropsychiatric manifestations, such as apathy, anhedonia, depression and anxiety. These hitherto generally neglected non-motor symptoms, have gained increasing interest in medical and scientific communities over the last decade because of the extent of their negative impact on PD patients' quality of life. Although recent clinical and functional imaging studies have provided useful information, the pathophysiology of apathy and associated affective impairments remains elusive. Our aim in this review is to summarize and discuss recent advances in the development of rodent models of PD-related neuropsychiatric symptoms using neurotoxin lesion-based approaches. The data collected suggest that bilateral and partial lesions of the nigrostriatal system aimed at inducing reliable neuropsychiatric-like deficits while avoiding severe motor impairments that may interfere with behavioral evaluation, is a more selective and efficient strategy than medial forebrain bundle lesions. Moreover, of all the different classes of pharmacological agents, D2/D3 receptor agonists such as pramipexole appear to be the most efficient treatment for the wide range of behavioral deficits induced by dopaminergic lesions. Lesion-based rodent models, therefore, appear to be relevant tools for studying the pathophysiology of the non-motor symptoms of PD. Data accumulated so far confirm the causative role of dopaminergic depletion, especially in the nigrostriatal system, in the development of behavioral impairments related to apathy, depression and anxiety. They also put forward D2/D3 receptors as potential targets for the treatment of such neuropsychiatric symptoms in PD.


Alpha-1-antitrypsin deficiency presenting as bronchiectasis.
We describe three patients with the clinical and radiographic features of progressive bronchiectasis, and without evidence of emphysema, associated with deficiency of alpha-1-antitrypsin. The association of hereditary alpha-1-antitrypsin deficiency and pulmonary emphysema is well recognized (Eriksson 1965). About half of these subjects also have chronic bronchitis as defined by persistent sputum production (Tobin et al. 1983). Bronchographically proven bronchiectasis without apparent emphysema has been reported in only one subject with alpha-1-antitrypsin deficiency (Longstretch et al. 1975).


Productivity of Agave deserti: measurement by dry weight and monthly prediction using physiological responses to environmental parameters.
An "environmental productivity index" based on physiological responses to three environmental variables was used to predict the net productivity of a common succulent perennial of the Sonoran Desert, Agave deserti, on a monthly basis. Productivity was also independently measured in the field from dry weight changes. The index was based on soil water availability, day/night air temperatures, and photosynthetically active radiation (PAR), which were individually varied in the laboratory and the effect on net CO2 uptake by the leaves determined. From monthly precipitation, temperature, and PAR at the field site together with the responses measured in the laboratory, an index (maximum value of unity) was assigned to each of these three environmental variables and their product was termed the environmental productivity index. This index indicates the fraction of maximal CO2 uptake expected in the field for each month (well-watered A. deserti assimilated 285 mmol CO2 m-2 leaf area day-1 at PAR saturation and optimal day/night temperatures of 25° C/15° C). The dry weight analysis was based on the monthly unfolding of new leaves from the central spike of the rosette and their seasonal increase in dry weight, which were determined in the field. The production of new leaves was highly correlated with the environmental productivity index (r2=0.93), which in turn was highly correlated with the water status index (r2=0.97). After correction for respiration by folded leaves, stem, and roots, plant productivity predicted by the average environmental productivity index (0.36) over a wet June-to-October period agreed within 4% with the productivity based on the conventional dry weight analysis. The net productivity of A. deserti over this 5-month period was 0.57 kg m-2 ground area (5.7 Mg ha-1), a large value for a desert CAM plant. The environmental productivity index proposed here may provide a reliable means for predicting net productivity on a monthly basis, which may be particularly useful for species in relatively variable environments such as deserts.


[Antigen-specific and antigen-nonspecific reactions of the immunity system in vaccination with a purified concentrated staphylococcal anatoxin].
Immunization with purified and concentrated staphylococcal toxoid leads to the rapid 12-fold rise of the level of anti-alpha-toxin and to the 17-fold rise of the titres of antibodies to extracellular staphylococcal products. At the period of immunization phasic changes in cell-mediated and humoral immunity characteristics, indicative of the state of the nonspecific resistance system, can be observed. These changes consist in the transient suppression of phagocytosis, including a decrease in the activity of intraleukocytic bactericidal systems (myeloperoxidase and cation protein), a decrease in the activity of the complement and the bactericidal activity of the blood serum, which should be taken into account when using this immune preparation for therapy.


The comparative value of noninvasive testing for diagnosis and surveillance of deep vein thrombosis.
This is a prospective analysis of 351 patients in two distinct groups undergoing ascending phlebography, impedance plethysmography (IPG), and/or phleborheography (PRG) within the same 24-hour period. One hundred twenty patients also had a 125I-fibrinogen uptake test (RFUT). The two patient groups consisted of the following: those patients evaluated because of suspicion of deep vein thrombosis (DVT) (diagnostic) and those patients at high risk for postoperative DVT (total joint replacement) who had routine noninvasive testing and ascending phlebography (surveillance). The overall sensitivities for IPG and PRG were significantly better in the diagnosis group (71% [69 of 97 patients] and 78% [82 of 105], respectively) compared with the surveillance group (20% [14 of 71] and 27% [17 of 63], respectively) (p less than 0.0001). The sensitivities for IPG and PRG detecting proximal (A/K) thrombi was 83% (68 of 82 patients) and 92% (79 of 86) in the diagnosis group compared with 32% (11 of 34) and 33% (9 of 27) in the surveillance group (p less than 0.0001). Although there was no difference in overall incidence of DVT between the diagnosis group (56%, 118 of 212 patients) and the surveillance group (55%, 76 of 139), the results can be explained by the difference in A/K thrombi (84% [99 of 118] and 47% [36 of 76]) (p less than 0.001) and occlusive A/K thrombi (84% [58 of 69] and 23% [7 of 31]) (p less than 0.0001), respectively. Of the patients with A/K thrombi, 97% (67 of 69) in the diagnosis group had hemodynamically detectable thrombi compared with only 48% (12 of 25) in the surveillance group (p less than 0.001). Combining the RFUT with the noninvasive studies for surveillance significantly improved the sensitivity for both A/K and distal thrombi. Patient selection also appears to have a significant influence on the results of the combination of IPG and RFUT when the current surveillance group is compared with similarly performed studies in a previously reported diagnosis group. The location and magnitude of thrombi in any patient population can be skewed depending on indications and timing of testing, thereby significantly affecting the sensitivity of noninvasive tests. IPG and PRG are reliable for evaluating patients with suspected DVT. However, patients with postoperative DVT have a high incidence of nonocclusive thrombi. Because noninvasive hemodynamic tests cannot identify accurately postoperative DVT, they cannot be used to generate epidemiologic data or as end points for studies evaluating efficacy of prophylaxis in patients undergoing total joint replacement, and anatomic studies of the deep venous system continue to be required.


Electron-microscopical study of the operculum in anuran tadpole after extirpation of the right forelimb during metamorphosis.
The process of histolysis and fenestration of the skin of the prospective opercular perforation region of Rana japonica after extirpation of the right forelimb was observed during metamorphosis by transmission and scanning electron microscopy. Epidermal cells of the belly of the tadpole, including the operculum, are extremely similar in their ultrastructure. Epidermal cells of the prospective opercular perforation region during metamorphosis become thin and vacuolated especially around the nucleus perhaps by autolysis, associated with lysosomal activity. The histolysis and formation of the perforation of the operculum occurs in the complete absence of forelimb. Macrophages containing phagosomes and lymphocytes or other blood cells are almost always found in the intercellular epidermis. Necrotic epidermal cells progressively separate by cleft formation and slough off without cornification.


Solid state 13C NMR study of collagen molecular dynamics in hard and soft tissues.
The molecular dynamics of the collagen backbone in intact connective tissues has been elucidated using 13C line shape analysis. Since one-third of the amino acid residues in collagen are glycines, we have labeled: (a) reconstituted lathrytic (uncross-linked) chick calvaria collagen fibrils; (b) rat tail tendon (cross-linked); and (c) rat calvaria (cross-linked and mineralized) collagen with [1-13C]glycine. The proton-enhanced and normal 90 degrees - t proton-decoupled spectrum of each collagen sample shows an asymmetric chemical shift powder pattern for the glycine carbonyl carbon. The powder line width, delta, (delta = sigma zz - sigma xx) at 22 degrees C for the uncross-linked reconstituted collagen fibril is 108 ppm, whereas the maximum value of delta (140 ppm) is observed for the cross-linked and mineralized collagen fibrils in rat calvaria. The powder line widths for the cross-linked fibrils in tail tendons and demineralized calvaria are 124 and 120 ppm, respectively. However, since the same line shape and line width (145 ppm) are observed for all samples at -35 degrees C, the difference in delta values observed at room temperature is attributed to differences in molecular mobility of collagen in various samples. The line shapes are analyzed using a dynamic model in which azimuthal orientation of the collagen backbone is assumed to fluctuate as a consequence of reorientation about the helix axis. The observed line shapes are sensitive to motions having correlation times less than approximately 10(-4) s and the analysis provides the values of the root mean square fluctuation in azimuthal angle, gamma rms, due to such motions. It is found that gamma rms equals 41 degrees, 33 degrees, and 14 degrees for the uncross-linked, cross-linked, and mineralized collagens, respectively. These results provide the first information about the extent that cross-linking and mineralization restrict molecular motion in collagen.


Branched-chain amino acid supplementation promotes aerobic growth of Salmonella Typhimurium under nitrosative stress conditions.
Nitric oxide (NO) inactivates iron-sulfur enzymes in bacterial amino acid biosynthetic pathways, causing amino acid auxotrophy. We demonstrate that exogenous supplementation with branched-chain amino acids (BCAA) can restore the NO resistance of hmp mutant Salmonella Typhimurium lacking principal NO-metabolizing enzyme flavohemoglobin, and of mutants further lacking iron-sulfur enzymes dihydroxy-acid dehydratase (IlvD) and isopropylmalate isomerase (LeuCD) that are essential for BCAA biosynthesis, in an oxygen-dependent manner. BCAA supplementation did not affect the NO consumption rate of S. Typhimurium, suggesting the BCAA-promoted NO resistance independent of NO metabolism. BCAA supplementation also induced intracellular survival of ilvD and leuCD mutants at wild-type levels inside RAW 264.7 macrophages that produce constant amounts of NO regardless of varied supplemental BCAA concentrations. Our results suggest that the NO-induced BCAA auxotrophy of Salmonella, due to inactivation of iron-sulfur enzymes for BCAA biosynthesis, could be rescued by bacterial taking up exogenous BCAA available in oxic environments.


The age gap: teaching students about health education for the elderly.
In learning about caring for elderly people, student nurses need to be aware of their attitudes to the elderly and also what they themselves perceive to be their health needs.


Distribution of insulin receptor-like immunoreactivity in the rat forebrain.
Previous studies have suggested that insulin may play a role in the hormonal regulation of neurotransmitter metabolisms within the central nervous system. In order to provide additional information to support this hypothesis, we examined the distribution of insulin receptors within the forebrain of adult male rats. Insulin receptors were localized by immunocytochemistry, using an antibody directed against the carboxy-terminus of the beta-subunit of the insulin receptor. The antibody specificity was tested by immunoprecipitation of brain insulin receptors with antiserum and the purity of the receptor-antibody preparation was determined using hormone binding-assays with radiolabeled insulin and insulin-like growth factor-l. Insulin receptor-like immunoreactivity was found in a widespread, but selective, distribution on neurons throughout the rat forebrain. Double-labeling with glial fibrillary acidic protein did not demonstrate any detectable insulin receptor-like immunoreactivity on glial cells. Areas with the highest density of insulin receptor-like immunoreactivity were found in the olfactory bulbs, hypothalamus and median eminence, medial habenula, subthalamic nucleus, subfornical organ, CA 1/2 pyramidal cell layer of the hippocampus and piriform cortex. Double-staining of hypothalamic sections with somatostatin and vasopressin antisera revealed insulin receptor-like immunoreactivity on a subpopulation of somatostatin neurons in the periventricular region and on vasopressin neurons in the supraoptic nucleus. A moderately dense insulin receptor-like immunoreactivity was observed in layers II-IV of cerebral cortex, medial amygdala, reticular thalamic nucleus, zona incerta, and preoptic and septal regions, whereas a low density of insulin receptor-like immunoreactive neurons was found in basolateral amygdala and most thalamic regions. The basal ganglia and most parts of the thalamus were almost devoid of insulin receptor-like immunoreactivity. Our findings provide morphological support for a direct action of insulin on selected regions of the rat forebrain and suggest that the insulin receptor may modulate synaptic transmission or the release of neurotransmitters and peptide hormones in the CNS.


Effects of anti-Ig reagents on T cell functions I. Activation of rabbit T cells by anti-allotype antibodies.
Rabbit lymphocytes were analyzed by flow microfluorometry, using anti-T cell and anti-Ig reagents. Rabbit T cells and cells expressing surface Ig (B cells) appeared to belong to distinct subpopulations which could be separated on the basis of their selective adherence to nylon wool columns or to anti-Ig-coated dishes. Using flow microfluorometry, no evidence was obtained for the expression of a allotypes (VH framework) on T cells. Separated lymphocyte populations were functionally characterized using an in vitro proliferation assay. B and T cells from rabbit spleen or peripheral blood responded in a differential fashion to B and T cell-specific mitogens and to anti-Ig antibodies. Although such T cells did not respond upon stimulation with anti-Ig antibodies alone, significant proliferation could be induced by simultaneous addition of anti-Ig and T cell growth factor. In addition, activated T cells, derived from lymph nodes of immunized rabbits, generated a proliferative response upon stimulation with anti-Ig reagents alone. The above-mentioned effects on T cells could be obtained using heterologous anti-Ig antibodies or isologous anti-allotype antibodies, directed either against a allotypes (VH framework) or against b allotypes (kappa light chain). Antibodies against the Fc portion of rabbit Ig or against irrelevant allotypic specificities were ineffective in triggering T cells. Fab fragments from anti-allotype antibodies were equally stimulatory for T cells as compared to intact IgG, indicating that cross-linking of Ig-like molecules is not a necessary requirement for anti-Ig-induced T cell activation.


Chronic Cerebral Ischemia: Where "Evidence-Based Medicine" Fails Patients.
The publication of the COSS (Carotid Occlusion Surgery Study) trial in 2011 concluded that the addition of external carotid to internal carotid (EC-IC) bypass to a medical regimen "did not reduce the risk of ipsilateral ischemic stroke at 2 years" for patients with symptomatic internal carotid artery occlusion. This has resulted in decreasing referrals for surgical management under the guise of "evidence-based" medicine. The conclusions drawn from a study can only be as good as the selected end points. COSS did not consider important end points such as the effects of long-term cerebral ischemia or recurrent debilitating transient ischemic attacks (TIAs). However, the study is often quoted as "proof" that EC-IC bypass has no role in the treatment of these patients. A middle-aged patient presented to a primary neurologist with left-sided weakness and 2 months of personality changes. Magnetic resonance imaging (MRI) showed small watershed infarcts in the right hemisphere. An angiogram showed right carotid occlusion. Perfusion scans confirmed hypoperfusion. The patient was managed medically. Without any further clinical stroke events, the patient progressed to complete hemispheric infarction on MRI over the next 5 months. The public perception of the results of large randomized clinical trials is significantly altered by their ubiquitous dissemination and broad generalization without adequate understanding of the details. Careful assessment of the methodology and end points of a trial are essential when applying the results for evidence-based medicine to individual patients. This patient would have been considered a medical "success" in COSS, but her outcome with surgical intervention will never be known.


[Drug's administration via feeding tubes: evaluation of practices in an intensive care unit of a Tunisian hospital].
Drugs' administration via feeding tubes is a potential source of iatrogenic events for the intensive care patients because of the problem of not adapted galenic forms. We analyzed the prescriptions of patients with enteral feedings to determine if the galenic forms were compatible with administration via feeding tubes. We also observed and analyzed the methods of drugs passage by nurses. We analyzed 30 prescriptions of patients with enteral feedings in the intensive care unit of Habib Bourguiba Sfax hospital, by a prospective and exhaustive way. We also, observed and evaluated the practices of preparation and administration of drugs to these patients via feeding tube by nurses. Only 12% of drugs were liquids. Eighty-eight percent of the drugs were pulverised and capsule open before administration. The galenic form was not in conformity for 20% of drugs because of the prohibition to crushing tablet or opening capsule (gastroresistant form was dissolved), or because of the administration of a parenteral form (risk of irritation). Among 78 drugs administered by 10 different nurses, the time between passage of the drug and enteral nutrition were not respected for 59% of the observations. The drugs were managed in mixture for 90% of the observations. The gloves were not worn in 80% of observations. No rinsing is made between consecutive administrations and before administration. This study shows that it is possible to reduce risk of administration errors in the intensive care unit and to facilitate the administration of drug via feeding tube by prescribing liquid oral form or soluble solid oral form. It also shows the need for cooperation with the pharmacist in order to adapt the galenic forms and to redact protocol of administration.


Development of a self-report format of ASSIST with university students.
The Alcohol, Smoking and Substance Involvement Screening Test (ASSIST) is a reliable and valid tool to early detection in the harmful and hazardous drug use in primary care settings when administered by interview in the general population. As the risk of substance related problems in university students is high, it is necessary to have screening instruments that can be used beyond the health care settings. Thus, we compared a self-report adaptation of ASSIST with the validated interview format in a convenience sample of university students. A counter-balance design was chosen with students alternating between the interview and the self-report formats. Both formats were completed by all students (n=170) over 30 days. The scores for total involvement, tobacco, alcohol, cannabis and cocaine obtained from the two formats demonstrated good intra-class correlation coefficient (ICC >0.60). The agreement assessed by kappa between questions of the two formats was considered moderate for tobacco (0.76) and cannabis (0.69) and discrete for alcohol (0.47). The consistency of the self-report questionnaire was also good to moderate (Cronbach's alpha of 0.90 for tobacco, 0.71 for alcohol, 0.86 for cannabis and 0.89 for cocaine) and showed acceptable sensitivity (66.7-100%) and specificity (83.5-97.1%) for tobacco, alcohol, cannabis and cocaine when compared to the ASSIST interview format (gold standard). The findings suggest that self-report version is as acceptable as the interview and that the scores on the two formats are comparable. However, the participants reported more motivation for change behavior and more concern about substance use when they were interviewed.


Increased expression of osteopontin in patients with triple-negative breast cancer.
Patients with triple-negative [oestrogen receptor (ER) negative, progesterone receptor (PR) negative, and human epidermal growth factor receptor 2 (HER-2/neu) negative] breast cancer, accounting for about 15% of breast cancer cases, are associated with aggressive histology, poor prognosis and shorter survival. Osteopontin is a chemokine-like phosphorylated glycoprotein that plays important role in cancer progression and is found to be a metastasis-associated protein in breast cancer. The goal of the study was to evaluate osteopontin protein expression levels in triple-negative breast carcinomas to determine if they correlated with clinicopathological parameters, thus providing additional support for osteopontin functioning and better understanding of triple-negative breast cancer. A database of 239 patients, in whom all three markers (ER, PR, and HER-2/neu) were available, was reviewed. We performed osteopontin protein expression analyses by means of immunohistochemistry on 117 breast carcinoma tissue samples, and then assessed the mean value of osteopontin expression against triple-negative status and clinicopathological parameters. Of the 239 patients in the study, 47 were classified as triple negative. Of the 117 osteopontin-test patients in this cohort, mean osteopontin levels were significantly higher in the triple-negative breast cancers than in non-triple-negative subtype (P = 0.035). TNM (tumours, nodes, metastases) stage were significantly associated with osteopontin levels (P = 0.038). Univariate analysis showed tumour cell osteopontin positivity above an optimized cut-point to be significantly associated with decreased disease-free survival, but not overall survival. In the multivariate model, osteopontin was an independent prognostic factor for disease-free survival. Patients with osteopontin overexpression develop predominantly triple-negative tumours. Osteopontin overexpression is associated with tumour aggressiveness and poor prognosis.


[Adaptation, validity, and reliability of the Metacognition Questionnaire-30 for the Turkish population, and its relationship to anxiety and obsessive-compulsive symptoms].
The present study aimed to evaluate the psychometric properties of the short form of the Metacognition Questionnaire (MCQ-30) in a population of Turkish university students. The metacognitions questionnaire (MCQ) measures individual differences in a selection of metacognitive beliefs, judgments, and monitoring tendencies considered important in the metacognitive model of psychological disorders. The study included 850 university students from 15 Turkish universities. Mean age age of the participants was 21.22 years, 282 (33.17%) were female, and 568 (66.82%) were male. Construct validity was evaluated by exploratory and confirmatory factor analysis (CFA). Exploratory factor analysis revealed that the Turkish version of MCQ-30 has 5 components, which is same factor structure as the original form. In addition, the fit indices of CFA suggested an acceptable fit to a 5-factor model consistent with the original MCQ-30. The Turkish MCQ-30 showed acceptable to good test-retest reliability, internal consistency, and convergent validity. Significant positive relationships between the subscales of MCQ-30 and measures of anxiety and obsessive-compulsive symptoms provided further support for the convergent validity of the Turkish version. Moreover, significant negative correlations were observed between age and the MCQ-30 subscales, and the effect of gender was significant on some of the subscales. The psychometric properties of the Turkish version of MCQ-30 showed that the instrument is a valuable additional tool for the assessment of metacognition in Turkey.


Stability of an RNA secondary structure affects in vitro transcription pausing in the trp operon leader region.
Transcription of the tryptophan (trp) operon of Escherichia coli and other bacterial species is regulated by the formation of alternative secondary structures in the leader segment of the transcript. During in vitro transcription of the trp leader region RNA polymerase pauses at base pair 92 after synthesis of an RNA hairpin secondary structure. We studied the dependence of pausing on hairpin stability by examining mutant trp templates containing base pair substitutions in the region corresponding to the hairpin secondary structure. Base changes that lower the stability of the hairpin were found to reduce both the frequency and half-life of RNA polymerase pausing while base changes that do not affect hairpin stability had little effect on pausing. Pausing was enhanced by the nusA protein; this enhancement was greatly reduced on mutant templates specifying less stable hairpins. The frequency of pausing on some mutant templates was correlated with the extent of read-through transcription beyond the trp attenuator, suggesting a possible role for pausing in the coupling of transcription and translation during transcription of the leader region of the operon.


Surgical gauze pseudotumor.
Gauze forgotten at operations can be potentially life threatening, but such cases are seldom reported because of the medicolegal implications. We have presented a series of seven patients with long-term surgical gauze retention, four after pelvic operations, one after cholecystectomy, one after laryngectomy, and one after mastectomy. The median time interval between operation and gauze removal was 5 years. In three patients this was diagnosed as a tumoral mass, in three as an intestinal occlusion, and in one the gauze was found incidentally. All foreign bodies were removed and no patient died. The clinical and diagnostic aspects of retained surgical gauze have been discussed and the need for radiopaque markers in them have been emphasized.


Contribution of cytochrome P450 metabolites to bradykinin-induced vasodilation in endothelial NO synthase deficient mouse hearts.
We have characterized the contribution of endothelial nitric oxide synthase (eNOS), cyclo-oxygenase (COX) and cytochrome P450 (CYP450) to the bradykinin (BK)- induced vasodilation in isolated hearts from wildtype (WT) and eNOS deficient mice (eNOS-/-). The endothelium-dependent vasodilation by bradykinin (BK, 1 microM) was significantly lower in eNOS-/- hearts than that in WT hearts (+247% and +325% of basal flow, respectively), while there was no difference in the endothelium-independent vasodilation by adenosine. In WT hearts, the BK-induced vasodilation was markedly attenuated following inhibition of NOS with ETU (10 microM) but not after COX inhibition with diclofenac (3 microM) (P<0.01). In line with this finding, Bk did not increase the cardiac prostacyclin release as measured by ELISA for 6-keto-PGF1alpha in the coronary venous effluent. In eNOS-/- hearts, the flow response to BK was insensitive to both NOS and COX inhibition. The NOS/COX-independent vasodilatory factor which remained under L-NMMA+DF application was almost completely eliminated by either clotrimazole (3 microM), miconazole (0.5 microM) or 17-ODYA (1 microM), suggesting that it was a metabolite of CPY450 enzymes. Sulfaphenazole (10 microM), a CYP450 2C inhibitor, exerted only a minimal inhibitory effect. In eNOS-/- hearts the effect of CYP450 inhibitors on the BK response was significantly more pronounced than in WT hearts, indicating an enhanced contribution of CYP450 enzymes. These findings suggest that in isolated mouse hearts the BK-induced vasodilation is mediated by NO and CYP450 metabolites but not by prostaglandins. The CYP450 dependent vasodilator was was functionally up-regulated in eNOS-/- hearts and thus likely to compensate for the loss of eNOS in the coronary circulation.


Structural characteristics of low-digestible sweet potato starch prepared by heat-moisture treatment.
The objectives of this study were to optimize the conditions for the preparation of sweet potato starch containing a high proportion of slowly digestible starch (SDS) by heat-moisture treatment (HMT), and to investigate the structural properties of SDS and resistant starch (RS) fractions isolated from this modified starch. Response surface methodology evaluation of considering temperature, time, and moisture content was conducted to determine the HMT condition for maximizing SDS yield. Maximum SDS content (50.9%) was obtained after 8.6 h at 113 °C with 22.8% moisture level. Scanning electron micrographs of RS fractions displayed growth ring structure and hollow regions. X-ray diffraction patterns of SDS + RS and RS fractions were not changed by HMT. Gelatinization enthalpy of the SDS + RS fraction was decreased more than that of the RS fraction. The results indicated that the SDS fraction consisted of less perfect crystallites and amorphous regions, whereas the RS fraction was composed mostly of crystalline regions.


Retinal nerve fiber layer volume measurements in healthy subjects using spectral domain optical coherence tomography.
To report retinal nerve fiber layer (RNFL) volume measurement using spectral domain optical coherence tomography (SD-OCT). We determined the characteristics and effecting factors of the RNFL volume in healthy eyes. Eighty-six eyes from 86 healthy volunteers were included. All participants underwent 3D-disc scanning using SD-OCT. We developed a computer program using Matlab software to extract RNFL thickness values from colored topographic RNFL thickness map. Total average, quadrant, and clock-hour RNFL volume were automatically calculated between 2.5 and 5 mm diameter circles using the length, width, and height of each pixel derived from the RNFL thickness map. The mean total RNFL volume was 1.478±0.124 mm and RNFL volume profiles showed double-hump patterns. RNFL volumes of superior and inferior quadrants were significantly greater than those of temporal and nasal quadrants (all P<0.001). A strong correlation existed between RNFL volume and conventional circumpapillary RNFL thickness measured by built-in software (R=0.895 to 0.972). Image quality and age were associated with RNFL volume measurement. RNFL volume measurement from RNFL thickness map analysis provides an alternative approach to characterizing and evaluating RNFL. RNFL volume measurement may be valuable for the objective and quantitative evaluation of 3-dimensional RFNL changes.


LncRNA MIAT Promotes Allergic Inflammation and Symptoms by Targeting MiR-10b-5p in Allergic Rhinitis Mice.
Allergic rhinitis (AR) is one of the most common noninfectious respiratory diseases caused by immunoglobulin E (IgE) response. The study sought to explore the relationship between lncRNA MIAT and miR-10b-5p and their interaction in the regulation of allergic phenotypes in allergic rhinitis (AR) mice. A mice model of AR was constructed using ovalbumin (OVA) sensitization. AR mice were treated with miR-10b-5p agomiR and LNA mediated lncRNA MIAT. The targeting relationship between MIAT and miR-10b-5p was analyzed by the ENCORI website and dual-luciferase reporter assay. The numbers of rubbing and sneezing of mice were counted. Hematoxylin-eosin (HE) staining visualized the eosinophils infiltration in nasal mucosa tissues of mice. The percentage of Th17 cells was quantitated by flow cytometry analysis. ELISA was used to detect the levels of serum OVA-specific IgE, the Th12 cytokine IL-4, and inflammatory cytokines (IL-6, IL-17). MIAT was up-regulated in the nasal mucosa of AR mice, while miR-10b-5p was down-regulated. MIAT directly suppressed miR-10b-5p expression in AR mice. The numbers of rubbing and sneezing, the percentage of Th17 cells, and the levels of OVA-specific IgE, IL-4, IL-6, and IL-17 in AR mice were decreased by miR-10b-5p overexpression, which was reversed by MIAT overexpression. The eosinophils infiltration in AR mice was inhibited by miR-10b-5p overexpression, which was also reversed by MIAT overexpression. The present study demonstrates that MIAT overexpression Promotes allergic inflammation and symptoms by activating Th17 immune response via miR-10b-5p inhibition.


Excess risk of mortality and cardiovascular events associated with smoking among patients with diabetes: meta-analysis of observational prospective studies.
Numerous studies have demonstrated that both smoking and diabetes are risk factors for mortality and caused-specific cardiovascular events. However, few studies systematically investigated to what extent the excess risk could be attributed to smoking among diabetic patients. Literature references were searched up to April 2011 in MEDLINE and EMBASE, supplemented by manual searches. Inclusion criteria were prospective cohort studies, assessment of the association between smoking and total mortality, cardiovascular death, incidence of coronary heart disease (CHD), stroke and myocardial infarction (MI) in diabetic patients. Of 3758 studies in the literature searched, 46 were eligible with approximately 130,000 diabetic patients. The relative risk (RR) comparing smokers with nonsmokers was 1.48[95% confidential interval (CI): 1.34-1.64] for total mortality (27 studies), 1.36(1.22-1.52) for cardiovascular mortality (9 studies), 1.54(1.31-1.82) for CHD (13 studies), 1.44(1.28-1.61) for stroke (9 studies) and 1.52(1.25-1.83) for MI (7 studies). Furthermore, the excess risk was observed among former and current smokers with a greater risk in current smokers. Subgroup analysis showed that the increased risk appeared to be consistent regardless of several study characteristics with the RRs ranging from 1.31 to 1.94 for all-cause mortality, 1.37 to 2.28 for CHD, 1.21 to 1.87 for stroke, 1.13 to 1.74 for cardiovascular mortality and 1.15 to 2.01 for MI. Smoking amplified the risk of mortality as well as cardiovascular events and the effect size for CHD appeared to be higher than other events in diabetic patients. Moreover, a trend of decreasing risk was observed among smoking quitters.