Staphylococcus δ-toxin induces allergic skin disease by activating mast cells.
Atopic dermatitis is a chronic inflammatory skin disease that affects 15-30% of children and approximately 5% of adults in industrialized countries. Although the pathogenesis of atopic dermatitis is not fully understood, the disease is mediated by an abnormal immunoglobulin-E immune response in the setting of skin barrier dysfunction. Mast cells contribute to immunoglobulin-E-mediated allergic disorders including atopic dermatitis. Upon activation, mast cells release their membrane-bound cytosolic granules leading to the release of several molecules that are important in the pathogenesis of atopic dermatitis and host defence. More than 90% of patients with atopic dermatitis are colonized with Staphylococcus aureus in the lesional skin whereas most healthy individuals do not harbour the pathogen. Several staphylococcal exotoxins can act as superantigens and/or antigens in models of atopic dermatitis. However, the role of these staphylococcal exotoxins in disease pathogenesis remains unclear. Here we report that culture supernatants of S. aureus contain potent mast-cell degranulation activity. Biochemical analysis identified δ-toxin as the mast cell degranulation-inducing factor produced by S. aureus. Mast cell degranulation induced by δ-toxin depended on phosphoinositide 3-kinase and calcium (Ca(2+)) influx; however, unlike that mediated by immunoglobulin-E crosslinking, it did not require the spleen tyrosine kinase. In addition, immunoglobulin-E enhanced δ-toxin-induced mast cell degranulation in the absence of antigen. Furthermore, S. aureus isolates recovered from patients with atopic dermatitis produced large amounts of δ-toxin. Skin colonization with S. aureus, but not a mutant deficient in δ-toxin, promoted immunoglobulin-E and interleukin-4 production, as well as inflammatory skin disease. Furthermore, enhancement of immunoglobulin-E production and dermatitis by δ-toxin was abrogated in Kit(W-sh/W-sh) mast-cell-deficient mice and restored by mast cell reconstitution. These studies identify δ-toxin as a potent inducer of mast cell degranulation and suggest a mechanistic link between S. aureus colonization and allergic skin disease.


Increased levels of lactate dehydrogenase and hypertension are associated with severe illness of COVID-19.
Coronavirus disease 2019 (COVID-19) is amid an ongoing pandemic. It has been shown that patients with cardiovascular comorbidities are at higher risk of severe illness of COVID-19. To find out the relationship between cardiovascular comorbidities and severe illness of COVID-19. The clinical data of 140 COVID-19 patients treated from January 22, 2020 to March 3, 2020 at our hospital were retrospectively collected. The clinical characteristics were compared between patients with mild illness and those with severe illness. There were 75 male patients and 65 female patients (53.6% vs 46.4%). The mean age was 45.4 ± 14.6 years (range, 2-85 years). Most of the patients had mild illness (n = 114, 81.4%) and 26 patients had severe illness (18.6%). The most common symptom was fever (n = 110, 78.6%), followed by cough (n = 82, 58.6%) and expectoration (n = 51, 36.4%). Eight patients were asymptomatic but were positive for severe acute respiratory syndrome coronavirus 2 RNA. Patients with severe illness were significantly more likely to be hypertensive than those with mild illness [(10/26, 38.4%) vs (22/114, 19.3%), P = 0.036]. The levels of lactate dehydrogenase were significantly higher in the severe illness group than in the mild illness group (299.35 ± 68.82 vs 202.94 ± 63.87, P < 0.001). No patient died in either the severe illness or the mild illness group. Hypertension and elevated levels of lactate dehydrogenase may be associated with severe illness of COVID-19.


(2E)-1-(2,5-Dimeth-oxy-phen-yl)-3-(3-nitro-phen-yl)prop-2-en-1-one.
In the title compound, C(17)H(15)NO(5), an intra-molecular C-H⋯O hydrogen bond generates an S(6) ring motif. The benzene rings form a dihedral angle of 6.45 (7)° with each other. In the crystal, inversion dimers linked by pairs of C-H⋯O hydrogen bonds generate R(2) (2)(8) loops. Adjacent dimers are further connected by C-H⋯O hydrogen bonds into an infinite chain along the [011] direction.


Evaluation of the reliability of preoperative descriptive airway assessment tests in prediction of the Cormack-Lehane score: A prospective randomized clinical study.
In this study we investigated and compared the predictive values of different airway assessments tests including thyromental height measurement test, which has been recently suggested, in difficult laryngoscopy (Cormack and Lehane [C-L] scores 3 and 4). In addition, we compared the effectiveness of methods and C-L scores, by IDS, in terms of predicting difficult intubation. Prospective, blinded study. Maltepe University. Four hundred fifty-one patients selected randomly who underwent general anesthesia. In this study we compared predictive value of thyromental height measurement test (TMH), which has been recently suggested, modified Mallampati test (MMT), upper lip bite test (ULBT), and thyromental distance measurement test (TMD) in difficult laryngoscopy. Final C-L scores were compared with intubation difficulty scale (IDS) in terms of predicting difficult intubation. Patient's American Society of Anesthesiology score, age and weight were recorded. TMH, TMD, MMT, ULBT, IDS and C-L scores were measured and determined. The optimal cut-off point for TMH for predicting difficult laryngoscopy was 43.5 mm and for TMD was 82.06 mm. Use of TMH <43.5 with MMT has the highest sensitivity for predicting difficult intubation (78.38) with 75.36% specificity and 97.50% negative predictive value. TMH showed sensitivity of 91.89% and specificity 52.17% at 50 mm cut-off value. In the comparison of the area under the receiver operating characteristic curve values, none of the tests came forth individually or in combination with MMT test. The present study demonstrates the practicality of TMH as a digitalized test however the clinical benefits of TMH in daily medical practice are drawn into question. The additional variable of race may have had some bearing on this and further studies, larger in patient sample size, may need to use different methodology concerning age-, sex-, and race-dependent variables in evaluating these tests.


Robust estimation of critical values for genome scans to detect linkage.
Estimation of study specific critical values for linkage scans (suggestive and significant thresholds) is important to identify promising regions. In this report, I propose a fast and concrete recipe for finding study specific critical values. Previously, critical values were derived theoretically or empirically. Theoretically-derived values are often conservative due to their assumption of fully informative transmissions. Empirically-derived critical values are computer and skill intensive and may not even be computationally feasible for large pedigrees. In this report, I propose a method to estimate critical values for multipoint linkage analysis using standard, widely used statistical software. The proposed method estimates study-specific critical values by using Autoregressive (AR) models to estimate the correlation between standard normal statistics at adjacent map points and then use this correlation to estimate study-specific critical values. The AR-based method is evaluated using different family structures and density of markers, under both the null hypothesis of no linkage and the alternative hypothesis of linkage between marker and disease locus. Simulations results show the AR-based method accurately predicts critical values for a wide range of study designs.


Marginal and internal fit of feldspathic ceramic CAD/CAM crowns fabricated via different extraoral digitization methods: a micro-computed tomography analysis.
The aim of this study was to compare the fit of feldspathic ceramic crowns fabricated via 3 different extraoral digitizing methods. Twelve maxillary first premolars were prepared and 36 single crowns were fabricated via 3 extraoral digitizing methods using a laboratory scanner (n = 12): (1) scanning the typodont (ST [control] group); (2) scanning the impression (SI group); (3) scanning the stone cast (SC group). Micro-computed tomography was used to calculate two-dimensional marginal-internal gap and the three-dimensional volumetric gap between the crowns and their corresponding dies. The measured gaps were divided into 6 location categories as follows: marginal gap (MG), finish line gap (FLG), axial wall gap (AWG), cuspal gap (CG), proximal transition gap (PTG), and central fossa gap (CFG). The correlation between each of the 3 extraoral digitizing methods and the adaptation status of the crown margins were also evaluated. The Wilcoxon signed-rank test, Spearman's rank test, and Chi-square test were used for data analysis (α = 0.05). The marginal gaps in the ST, SI, and SC groups differed significantly (24, 198 and 117.6 µm, respectively) (p < 0.05). Significant differences were found between the groups with regard to internal gap measurements, with SI representing higher gap measurements at FLG, PTG and CFG locations (p < 0.05). 3D volumetric gap measurements did not differ significantly (p > 0.05). Under-extended margins observed in the SI and SC groups were correlated with the digitizing method (Cramer's V-square: 0.14). When performing extraoral digitalization, clinicians should choose to scan the stone cast as scanning the stone cast resulted in better internal and marginal fit compared to scanning the impression.


[Role of psychological correction in the combination treatment of patients with gastroesophageal reflux disease].
To evaluate the efficiency of psychotherapeutic methods on the clinical course of gastroesophageal reflux disease (GERD), quality of life (QL), esophagogastroduodenoscopic findings and 24-hour pH monitoring readings in patients with this condition. Sixty patients with GERD were divided into equal groups according to performed therapy: standard drug treatment or its combination with psychotherapeutic methods. The investigators estimated the degree of esophageal mucosal damage by esophagogastroduodenoscopy, esophageal acidity by 24-hour pH monitoring, and the magnitude of clinical manifestations by the Likert scale and assessed QL by the SF-36 questionnaire. The psychoemotional health component was analyzed using the clinical questionnaire for the identification and evaluation of neurotic states (CQIENS), the Leonhard-Shmichek characterological questionnaire, and the individual typological questionnaire. The patients with GERD were found to have individual personal traits and a definite psychological response. Incorporation of psychotherapeutic methods into the combination therapy was ascertained to have an additional therapeutic effect against the clinical manifestations of GERD, psychological responses in the patients, and QL indicators. There was a direct relationship between the psychological component of QL and the data of the CQIENS questionnaire. The most significant correlation was found between the CQIENS anxiety scores and the psychological component of QL (r = 0.73; p = 0.00004). Correlation analysis in the patients with GERD showed a relationship between the CQIENS anxiety scores and the clinical index (r = -0.68; p = 0.0001). The use of psychotherapeutic methods in addition to standard drug therapy improves both the physical and psychological states in patients with GERD and has some impact on a reduction in the magnitude of clinical symptomatology. This may contribute to the prompter recovery of patients and create prerequisites for improving their QL.


Linkage disequilibrium in laboratory strains of Drosophila ananassae is due to drift.
Two inversions in the opposite arms of the third chromosome are common in natural populations of Drosophila ananassae. When the strains are taken to laboratory, these inversions have been found to persist for several generations. Four natural populations were sampled and the data on the combinations between 3L and 3R karyotypes were obtained. For all the four populations, laboratory stocks (mass culture and isofemale lines) were established and maintained in food bottles by transferring fifty flies in each generation. After ten generations, chromosomes of all stocks were sampled. The results show that the two linked inversions are associated randomly in natural populations and in laboratory stocks established from mass culture. On the other hand, the same two inversions show non-random association (linkage disequilibrium) in several isofemale lines. The random association between inversions in natural and mass culture laboratory populations as contrasted to laboratory populations established from single females suggest that random drift is the cause of non-random association in isofemale lines. The tight linkage between the two inversions as evidenced by the results of recombination studies reported earlier, supports the notion that linkage disequilibrium is caused by drift.


Effect of Al substitution on the magnetization reversal and complex magnetic properties of NiCr2O4 ceramics.
A series of polycrystalline NiCr2-xAlxO4 (0.1 ≤ x ≤ 0.25) spinel ceramics have been synthesized using a sol-gel method. DC magnetization measurements are carried out at different temperatures and magnetic fields. A novel magnetization reversal has been observed in the field cooling process for the x = 0.2 sample, which can be ascribed to the competition between two magnetic sublattices due to their different temperature dependences. The magnetic interaction evolution, related to the complex magnetic properties, is revealed by exchange constants that have been estimated according to ferrimagnetic Curie-Weiss fitting and mean field theory. The fitting result confirmed the evolution of antiferromagnetic and ferromagnetic components with Al substitution, which is supported by the observations from the isothermal magnetization measurements. The positive and negative values of the magnetic moment can be utilized for storage applications based on the results of magnetic switching effect measurements.


The use of r-hFSH in treatment of idiopathic male factor infertility before ICSI.
To evaluate the efficacy of pre-treatment of idiopathic oligozoospermic patients with r-hFSH to improve the clinical results of ICSI. 82 infertile couples due to male factor who attended our center were included in the study. Thirty-six were randomized to the treatment group (group A) and forty-six to the control group (group B). The male partners in group A were treated with recombinant human FSH (r-hFSH; Gonal F®) 150 IU subcutaneously three times a week for a 3-months period. The control group (group B) did not receive any treatment. After the treatment couples of both groups underwent a cycle of ICSI. The fertilization rate was comparable in both groups. However, in the treatment group (group A), the clinical pregnancy rate was significantly higher (42%) compared to the control group (group B) (20%) (p < 0.02). Also, the implantation rate was significantly higher in treatment group (26%) compared to the control (15%) (p < 0.04). Miscarriage rate was lower (15.7%) in the treatment group than in the control (43.7%), and this difference was statistically significant (p < 0.05). Treatment of idiopathic male factor infertility with r-hFSH before ICSI improves clinical pregnancy rate, increases implantation rate and decreases the early pregnancy loss.


Activity of Paraoxonase/Arylesterase and Butyrylcholinesterase in Peripheral Blood of Gulf War Era Veterans With Neurologic Symptom Complexes or Post-Traumatic Stress Disorder.
Two groups of Gulf War era veterans, one exhibiting blurred vision, balance problems/dizziness, tremors/shaking, and speech difficulty and a second group with post-traumatic stress disorder (PTSD), but not the neurologic syndrome, were assessed for organophosphate-detoxifying enzyme paraoxonase/arylesterase (PON1) and its Q/R isoforms, butyrylcholinesterase (BuChE) and its U/A isoforms and cytokines. Defibrinated peripheral blood was evaluated for enzymes and cytokines. Trends toward elevation of Th2 cytokines interleukin-4 (IL-4) and IL-13 were observed in subjects with neurologic syndrome. Neither the activities nor isoforms of the enzyme, the neurologic symptoms, nor PTSD had any relationship to wartime deployment to the theater of combat. The negative outcomes described above suggest that exposure to organophosphates or other agents normally detoxified by PON1 and BuChE may not have contributed significantly to neurologic components of Gulf War Illness.


Inhibition of FGF receptor activity in glioma implanted into the mouse brain using the tetracyclin-regulated expression system.
We have investigated growth and vascularization of malignant glioma in mice upon conditional inhibition of fibroblast growth factor (FGF) receptor activity. C6 rat glioma cells were transfected with a dominant-negative fibroblast growth factor receptor-2 (FGFR2-DN) cDNA under the control of a tetracycline-regulated expression promoter (tet off) and implanted in the brain of immunodeficient mice. Magnetic resonance imaging analysis showed a significant decrease in tumor growth 14 days after implantation when FGFR2-DN was expressed compared to control. This size difference disappeared after 20 days. However, after 20 days, tumor and endothelial cells apoptosis were higher in the FGFR2-DN group and consequently angiogenesis was decreased whereas tumor cells were similarly associated with blood vessels at the tumor periphery. Pericyte coverage was not different between the two groups but a higher amount of pericytes not associated with vessels was found in the FGFR2-DN expressing group. This demonstrates, that conditional expression of inhibitor of FGF receptor activity in gliomas implanted in the brain of immunodeficient mice can be achieved efficiently, and that FGFs are major players in glioma development and in glioma angiogenesis.


Loss of function of axonemal dynein Mdnah5 causes primary ciliary dyskinesia and hydrocephalus.
Primary ciliary dyskinesia (PCD), also known as Kartagener's syndrome, is a human syndrome that results from ciliary dysfunction. This syndrome is characterized by recurrent respiratory infections, situs inversus and infertility. In some cases, hydrocephalus is also observed. We have characterized an insertional mutation in a mouse axonemal dynein heavy chain gene (Mdnah5) that reproduces most of the classical features of PCD, including recurrent respiratory infections, situs inversus and ciliary immotility. These mice also suffer from hydrocephalus and die perinatally. Electron microscopic studies demonstrate the loss of axonemal outer arms. These results show that mutations in Mdnah5 are a primary cause of PCD and provide direct evidence that mutations in an axonemal dynein can cause hydrocephalus. Mutations in the human DNAH5 have recently been identified in PCD patients. Comparison of the mouse model and the human data suggests that the degree of ciliary dysfunction is causally related to the severity of human PCD, particularly the presence of hydrocephalus.


Etoposide improves survival in high-grade glioma: a meta-analysis.
The purpose of this meta-analysis was to evaluate the therapeutic efficacy of topoisomerase inhibitors in the treatment of high-grade gliomas (HGGs). Using median overall survival (mOS) and survival gain, we compared the efficacy of chemotherapy drugs in a meta-analysis of 624 HGG studies, including 44,850 patients from studies published between 1976 and 2011. Patient cohorts treated with etoposide had significant improvement in mOS (15.66 months vs. 13.27 months, p=0.026, 49 vs. 795 cohorts) and significant survival gain advantage (p=0.022) over cohorts treated without etoposide. In contrast, patient cohorts treated with irinotecan had significantly worse mOS (10.20 vs. 13.55 months, p=0.008, 35 vs. 810 cohorts) and a disadvantage compared to cohorts treated without irinotecan in survival gain analysis. Results from this analysis suggest that etoposide may improve overall survival for patients with HGG, whereas the use of irinotecan might result in inferior outcomes.


Comparison of the haemodynamic profiles of the hypotensive action of verapamil and carvedilol.
Comparative haemodynamic investigations with carvedilol and verapamil were carried out on conscious, instrumented dogs. Arterial blood pressure, right atrial pressure (RAP), cardiac output (CO), heart rate, stroke volume and total peripheral resistance (TPR) were determined after intravenous (i.v.) injection of incremental doses (0.01-3 mg/kg) of the drugs. Carvedilol reduced the blood pressure in a dose-dependent manner, concomitant with a reduction in TPR. The RAP and the CO were not affected, indicating that arterial vasodilatation was induced by carvedilol. Verapamil showed a decrease in the blood pressure with a reduction of CO and SV. Moreover, at high doses the RAP was increased, indicating a reduction of the cardiac performance. Thus, in our experimental model remarkable differences between the haemodynamic effects of i.v. injections of carvedilol and verapamil have been observed, whereas after oral administration blood pressure also decreases after verapamil due to a reduction of TPR.


Migrant farmworker mothers talk about the meaning of food.
To describe the meaning of food in the family life of migrant farmworker mothers; specifically to describe their understanding of the relationship of dietary intake to health, the environmental contributors to their families' dietary intake, and the use of foods in the commemoration of family occasions. The PRECEDE-PROCEED self-management model guided the qualitative research study, which used short, guided interviews conducted at Midwest migrant camps. A sample of 43 Mexican mothers answered three interview questions, the answers to which were transcribed verbatim, read, sorted, and arranged into categories and then themes. Thematic analysis was used to analyze the data. The themes identified from the interviews were a) meal cost, b) lack of preparation time, c) distance to store locations, d) the importance of traditional ethnic foods, and e) limited knowledge of healthy food choices. Pediatric nurses should remember that before they dispense nutritional anticipatory guidance, situational and cultural assessments are needed. Because vulnerable populations such as migrant farmworkers have limited resources to follow nurses' teachings, nurses need to become knowledgeable about what is available in the community to support these families. In addition, facility appointment times may need to be expanded to accommodate their long work hours, and modes of transportation need to be evaluated. Recommendations for anticipatory guidance, prescriptions, treatments, and over-the-counter items need to consider cost and ease in purchase, and cultural acceptability.


Spiritual, religious, and existential concerns of cancer survivors in a secular country with focus on age, gender, and emotional challenges.
The purpose of this study was to examine the associations between self-reported spiritual/religious concerns and age, gender, and emotional challenges among cancer survivors who have completed a 5-day rehabilitation course at a rehabilitation center in Denmark (the former RehabiliteringsCenter Dallund (RC Dallund)). The data stem from the so-called Dallund Scale which was adapted from the NCCN Distress Thermometer and comprised questions to identify problems and concerns of a physical, psychosocial, and spiritual/religious nature. Descriptive statistics were performed using means for continuous variables and frequencies for categorical variables. Odds ratios were calculated by logistic regression. In total, 6640 participants filled in the questionnaire. Among participants, 21% reported one or more spiritual/religious concerns, the most reported concerns related to existence and guilt. Having one or more spiritual/religious concerns was significantly associated with age (OR 0.88), female gender (OR 1.38), and by those reporting emotional problems such as being without hope (OR 2.51), depressed (OR 1.49), and/or anxious (OR 1.95). Among participants, 8% stated they needed help concerning spiritual/religious concerns. Cancer patients, living in a highly secular country, report a significant frequency of spiritual/religious and existential concerns. Such concerns are mostly reported by the young, female survivors and by those reporting emotional challenges. Spiritual/religious and existential concerns are often times tabooed in secular societies, despite being present in patients. Our results call for an increased systemic attention among health professionals to these concerns, and a particular focus on identifying and meeting the spiritual/religious and existential concerns of women, the young and those challenged by hopelessness, depression, and anxiety.


Treatment of transthyretin familial amyloid polyneuropathy with tafamidis: a case report.
Familial amyloid polyneuropathy (FAP) is a rare hereditary disorder caused by mutations in the transthyretin (TTR) gene. Tafamidis is a TTR stabilizer able to prevent TTR tetramer dissociation, and several studies have demonstrated its safety and efficacy at slowing the progression of neuropathy in FAP caused by the TTR Val30Met mutation. However, nerve conduction study (NCS) and electromyography (EMG) results have yet to be reported in relation to FAP progression during tafamidis therapy. A 71-year-old man was admitted to the hospital because of severe numbness and walking difficulties. He did not complain of any autonomic dysfunction or visual disturbance, and he had no family history of neuromuscular disorders. A NCS and EMG indicated length-dependent axonal sensorimotor polyneuropathy. A sural nerve biopsy revealed amyloid deposits, and genetic testing demonstrated a TTR Val30Met mutation. We diagnosed the patient with FAP and treated him using tafamidis therapy. One year later, the patient is still on tafamidis therapy, and his symptoms have shown no significant change. The NCS showed no changes in the compound muscle action potential amplitudes of the left ulnar nerve, while EMG showed the fibrillation/positive sharp wave to have disappeared from the patient's upper limb. We conclude from these findings a slowing of the neurologic progression. Tafamidis was effective in slowing the neurologic progression over one year in a FAP patient with the TTR Val30Met mutation, and NCS and EMG were useful for assessing this therapeutic effect.


Seed Cryopreservation and Germination of Rhus glabra and the Critically Endangered Species Rhus michauxii.
Rhus michauxii is a perennial rhizomatous shrub native to the southeastern United States that is found mainly in sunny, dry, open rocky or sandy woodlands. Moreover, it is found on ridges or river bluffs in the inner coastal plane and lower piedmont of Virginia, Georgia, and the Carolinas. Habitat conversion to agriculture, suppression of fires, and low reproduction have caused R. michauxii to become rare and it is now federally listed as threatened. Methods are needed to multiply and conserve R. michauxii. Protocols were developed for seed cryopreservation, in vitro germination, and micropropagation for R. glabra and R. michauxii. Seed scarification in concentrated sulfuric acid for 6 h and germination on ½ MS medium resulted in germination up to 96% for control and cryopreserved seeds of R. glabra and 70 and 40% for control and cryopreserved seeds of R. michauxii. Shortly after germination in vitro, young seedlings were established in a greenhouse potting mix providing new plants from the endemic Georgia R. michauxii populations. Several of the findings meet goals within the R. michauxii recovery plan by providing methods for sexual and asexual multiplication and long-term seed storage under cryogenic conditions. The protocols developed will assist in the safeguarding and conservation of dwindling natural R. michauxii populations.


Exploring the impact of efficacy messages on cessation-related outcomes using Ecological Momentary Assessment.
Observational studies indicate that cigarette package inserts with efficacy messages about the benefits of quitting (i.e. response efficacy) and recommendations for successful cessation increase smokers' self-efficacy to quit and promote sustained cessation. However, the effects of inserts with such efficacy messages have not been studied using experimental designs. This study used ecological momentary assessment (EMA) to assess smokers' responses to efficacy inserts. In a randomized case-crossover study among smokers from the United States (n=23), participants received a one-week supply of cigarettes with efficacy inserts and a one-week supply without any inserts, and were randomized to use the packs with inserts on either the first or second week of the study. For 14 consecutive days, participants used a smartphone to answer brief surveys on cessation-related variables during smoking sessions and at the beginning of each day. Multilevel mixed-effects linear and logistic regression models compared responses during the insert period to those of the non-insert period. The insert period was associated with greater desire to quit (b=0.21, p=0.012), motivation to quit (b=0.18, p=0.001), self-efficacy to cut down (b=0.26, p<0.001) and to quit (b=0.28, p<0.000), and response efficacy/perceived benefits of quitting (b=0.13, p=0.012). Insert exposure was not significantly associated with forgoing cigarettes (OR=1.9, p=0.2). Results from this EMA study suggest that inserts with efficacy messages may promote determinants of smoking cessation. This is consistent with observational research in Canada, which is the only country to use inserts with efficacy messages as well as pictorial warnings about smoking risks on the outside of packs. Future studies should assess the extent to which efficacy inserts can not only be used to communicate health information to smokers but also work in synergy with pictorial warnings.


Isoflurane post-conditioning protects against intestinal ischemia-reperfusion injury and multiorgan dysfunction via transforming growth factor-β1 generation.
This study examined volatile anesthetic-mediated protection against intestinal ischemia-reperfusion injury (IRI). Intestinal IRI is a devastating complication in the perioperative period leading to systemic inflammation and multiorgan dysfunction. Volatile anesthetics, including isoflurane, have anti-inflammatory effects. We aimed to determine whether isoflurane, given after intestinal ischemia, protects against intestinal IRI and the mechanisms involved in this protection. : After IACUC approval, mice were anesthetized with pentobarbital and subjected to 30 minutes of superior mesenteric artery ischemia, followed by 4 hours of equianesthetic doses of pentobarbital or isoflurane. Five hours after reperfusion, small intestine tissues were analyzed for morphological injury, apoptosis, neutrophil infiltration, proinflammatory mRNAs, and TGF-(Transforming Growth Factor-)β1 levels. We also assessed hepatic and renal injury after intestinal IRI. Intestinal IRI with pentobarbital led to significant small intestinal dysfunction with increased mucosal injury, TUNEL (transferase biotin-dUTP nick end-labeling)-positive cells, neutrophil infiltration, and proinflammatory mRNAs as well as elevated plasma alanine aminotransferase and creatinine levels. Isoflurane exposure after IRI led to significant attenuation of intestinal, hepatic, and renal injuries. Furthermore, the protective effects of isoflurane were abolished by treatment with a TGF-β1 neutralizing antibody before induction of IRI. Finally, isoflurane exposure led to increased TGF-β1 levels in intestinal epithelial cells and in plasma. Our findings demonstrate that isoflurane post-conditioning protects against small intestinal injury and hepatic and renal dysfunction after severe intestinal IRI via induction of intestinal epithelial TGF-β1. Our findings support therapeutic applications of volatile anesthetics during the intraoperative and postoperative periods and imply an important role of TGF-β1 signaling in modulating multiorgan injury.


Two modeling strategies for empirical Bayes estimation.
Empirical Bayes methods use the data from parallel experiments, for instance observations Xk ~ 𝒩 (Θ k , 1) for k = 1, 2, …, N, to estimate the conditional distributions Θ k |Xk . There are two main estimation strategies: modeling on the θ space, called "g-modeling" here, and modeling on the×space, called "f-modeling." The two approaches are de- scribed and compared. A series of computational formulas are developed to assess their frequentist accuracy. Several examples, both contrived and genuine, show the strengths and limitations of the two strategies.


A single nucleotide polymorphism of AIRE gene located in the 21q22.3 increases the risk of rheumatoid arthritis.
Several studies addressed the association of autoimmune regulator (AIRE) gene polymorphism with the risk of rheumatoid arthritis (RA); however, their conclusions were inconsistent. For better investigating the effects of this polymorphism on the risk of RA, we conducted this study to evaluate the role of AIRE rs2075786 polymorphism in the risk of RA. Four eligible studies involving 6,755 cases and 7,970 controls were identified by searching the databases of PubMed, CNKI and EMBASE up to February 2017. Our study revealed that AIRE rs2075786 polymorphism was associated with an increased risk of RA under all genetic models. In the subgroup analysis, AIRE rs2075786 polymorphism contributed to RA susceptibility among Asians, but not among Caucasians. To summarize,, this meta-analysis confirms that AIRE rs2075786 polymorphism may play a significant role in increasing the risk of RA. Stratification analysis by ethnicity reveals that AIRE rs2075786 polymorphism is associated with an increased risk of RA among Asians, but not among Caucasians. These findings need further validation in the large multicenter case-control studies.


Accumulation of Chlorophyll, Chloroplastic Proteins, and Thylakoid Membranes during Reversion of Chromoplasts to Chloroplasts in Citrus sinensis Epicarp.
In vitro culture of pericarp segments from fruit of Citrus sinensis (L.) Osbeck cv Valencia was used to determine the temporal sequence in development of chloroplasts from chromoplasts during regreening of the epicarp. Regreening of chromoplasts closely resembled greening of etioplasts, except that regreening proceeded much more slowly. Chlorophyll, the light-harvesting chlorophyll a/b binding protein of photosystem II, the chlorophyll a binding protein of reaction center P-700 of photosystem I, thylakoid membranes, and adenosine triphosphate synthetase were all detected at very low levels in degreened epicarp. All of these increased in parallel during regreening of the epicarp. Ribulose 1,5-bisphosphate carboxylase (RuBPCase) levels were high in degreened epicarp and declined for the first 10 days of culture before reaccumulating in the regreening segments. Light was necessary for the accumulation of all of the chloroplastic components. A lack of exogenous nitrogen did not prevent the accumulation of any chloroplastic component except Ru-BPCase, although accumulation of the other components was reduced. Sucrose at 150 millimolar in media lacking nitrogen markedly inhibited the accumulation of chlorophyll and light-harvesting chlorophyll a/b-protein.


[Clinical studies of 16 cases of carcinomatous pericarditis].
We studied the effects of local therapy on cardiac tamponade caused by carcinomatous pericarditis in 16 cases. Five out of 16 cases (drainage group) received treatment by only drainage of pericardial effusion, and the remaining 11 cases (infusion group) had local infusion of antineoplastic agents into the pericardium after drainage. The median survival time was 16 weeks in the infusion group and 19 weeks in the drainage group. There was no statistically significant difference. Reaccumulation of pericardial effusion was recognized in 3 patients in the drainage group (60%), and in 3 patients in the infusion group (18%). Our conclusion was that local infusion had no effect on prolongation of the survival time, but had a suppressive effect on reaccumulation of pericardial effusion.


Mammalian Epidermis: A Compendium of Lipid Functionality.
Mammalian epidermis is a striking example of the role of lipids in tissue biology. In this stratified epithelium, highly specialized structures are formed that leverage the hydrophobic properties of lipids to form an impermeable barrier and protect the humid internal environment of the body from the dry outside. This is achieved through tightly regulated lipid synthesis that generates the molecular species unique to the tissue. Beyond their fundamental structural role, lipids are involved in the active protection of the body from external insults. Lipid species present on the surface of the body possess antimicrobial activity and directly contribute to shaping the commensal microbiota. Lipids belonging to a variety of classes are also involved in the signaling events that modulate the immune responses to environmental stress as well as differentiation of the epidermal keratinocytes themselves. Recently, high-resolution methods are beginning to provide evidence for the involvement of newly identified specific lipid molecules in the regulation of epidermal homeostasis. In this review we give an overview of the wide range of biological functions of mammalian epidermal lipids.


Dental occlusion in a split Amazon indigenous population: genetics prevails over environment.
Studies examining human and nonhuman primates have supported the hypothesis that the recent increase in the occurrence of misalignment of teeth and/or incorrect relation of dental arches, named dental malocclusion, is mainly attributed to the availability of a more processed diet and the reduced need for powerful masticatory action. For the first time on live human populations, genetic and tooth wear influences on occlusal variation were examined in a split indigenous population. The Arara-Iriri people are descendants of a single couple expelled from a larger village. In the resultant village, expansion occurred through the mating of close relatives, resulting in marked genetic cohesion with substantial genetic differences. Dental malocclusion, tooth wear and inbreeding coefficient were evaluated. The sample examined was composed of 176 individuals from both villages. Prevalence Ratio and descriptive differences in the outcomes frequency for each developmental stage of the dentition were considered. Statistical differences between the villages were examined using the chi-square test or Fisher's exact statistic. Tooth wear and the inbreeding coefficient (F) between the villages was tested with Mann-Whitney statistics. All the statistics were performed using two-tailed distribution at p≤0.05. The coefficient inbreeding (F) confirmed the frequent incestuous unions among the Arara-Iriri indigenous group. Despite the tooth wear similarities, we found a striking difference in occlusal patterns between the two Arara villages. In the original village, dental malocclusion was present in about one third of the population; whilst in the resultant village, the occurrence was almost doubled. Furthermore, the morphological characteristics of malocclusion were strongly different between the groups. Our findings downplay the widespread influence of tooth wear, a direct evidence of what an individual ate in the past, on occlusal variation of living human populations. They also suggest that genetics plays the most important role on dental malocclusion etiology.


Comparison of the differentiation of dental pulp stem cells and periodontal ligament stem cells into neuron-like cells and their effects on focal cerebral ischemia.
Recent studies have reported an increasing incidence of ischemic stroke, particularly in younger age groups. Dental pulp stem cells (DPSCs) and periodontal ligament stem cells (PDLSCs) are the most common stem cells acquired from the teeth of adults, even elderly people. However, there are no detailed reports on whether DPSCs or PDLSCs are suitable for the treatment of ischemic stroke. In this study, the in vitro differentiation of DPSCs and PDLSCs into neuron-like cells was evaluated. Then, we established a rat model of cerebral ischemia. DPSCs or PDLSCs were administered to animals, and the therapeutic effects of these two types of cells were investigated. The results showed that PDLSCs had a higher differentiation rate than DPSCs. Immunofluorescence studies showed that the expression of the neuronal differentiation marker Thy-1 was higher in PDLSCs than in DPSCs, and other gene markers of neuronal differentiation showed corresponding trends, which were confirmed by western blot analysis. In this process, the Notch and Wnt signaling pathways were inhibited and activated, respectively. Finally, rats with transient occlusion of the right middle cerebral artery were used as a model to assess the therapeutic effect of PDLSCs and DPSCs on ischemia. The results showed that rats in the PDLSC-treated group emitted significantly greater red fluorescence signal than the DPSC-treated group. PDLSC transplantation promoted the recovery of neurological function more effectively than DPSC transplantation. Hence, PDLSCs represent an autogenous source of adult mesenchymal stem cells with desirable biological properties and may be an ideal candidate for clinical applications.


Fabrication of Smart Tantalum Carbide MXene Quantum Dots with Intrinsic Immunomodulatory Properties for Treatment of Allograft Vasculopathy.
MXene nanomaterials have sparked significant interest among interdisciplinary researchers to tackle today's medical challenges. In particular, colloidal MXene quantum dots (MQDs) offer the high specific surface area and compositional flexibility of MXene while providing improvements to aqueous stability and material-cell interactions. The current study for the first time reports the development and application of immunoengineered tantalum-carbide (Ta4C3T x ) MQDs for in vivo treatment of transplant vasculopathy. This report comes at a critical juncture in the field as poor long-term safety of other MXene compositions challenge the eventual clinical translatability of these materials. Using rational design and synthesis strategies, the Ta4C3T x MQDs leverage the intrinsic anti-inflammatory and antiapoptotic properties of tantalum to provide a novel nanoplatform for biomedical engineering. In particular, these MQDs are synthesized with high efficiency and purity using a facile hydrofluoric acid-free protocol and are enriched with different bioactive functional groups and stable surface TaO2 and Ta2O5. Furthermore, MQDs are spontaneously uptaken into antigen-presenting endothelial cells and alter surface receptor expression to reduce their activation of allogeneic T-lymphocytes. Finally, when applied in vivo, Ta4C3T x MQDs ameliorate the cellular and structural changes of early allograft vasculopathy. These findings highlight the robust potential of tailored Ta4C3T x MQDs for future applications in medicine.


Screening strategies to identify new antibiotics.
The emergence of multi-drug resistant bacteria is one of the most critical medical problems currently facing humankind, which will only get worse if no new antibacterial drugs are launched. This article will first review commonly used screening strategies used to identify potential new antibiotics and then discuss novel screening methods. In addition, new assays, methods, biological targets and compounds with novel modes of action undergoing pre-clinical or clinical development are briefly discussed.


[Care of hospitalized adults: conceptual systems as the basis of activities].
The study analyze the social representation of the care attributes. Interpretation schemes were utilized to analyse the speeches of graduate course students of Adult Health Care Area. The results show a conceptual framework logical, with peculiar taxonomic relations.


Expression of the human IL-2 receptor on lymphocytes involves rapid turnover of its p55 alpha-subunit (Tac).
Upon mitogenic stimulation, both mRNA encoding the p55 alpha-subunit (Tac) of the human IL-2R alpha and IL-2R alpha protein are induced and expressed in tonsil lymphocyte populations over several days. Using a quantitative dot-blot immunoassay for the IL-2R alpha subunit, a rapid disappearance of this polypeptide from cells is demonstrated in the presence of the translation inhibitor, cycloheximide. The half-life of IL-2R alpha subunit protein is 2 to 3 h. This decline in cell-associated IL-2R alpha subunit is matched by a rapid decline in IL-2R alpha on the cell surface and is not accompanied by any increase in soluble IL-2R alpha protein. Long term expression of the IL-2R on the cell surface is thus the result of continual synthesis and rapid breakdown of IL-2R alpha chains in the cell. Steady state expression of the IL-2R after an immune stimulus hence depends upon continuous expression of the IL-2R alpha subunit gene. Rapid turnover of the unstable alpha-subunit on the cell surface provides a novel mechanism for sensitive control of functional IL-2R expression.


Neighbourhood socioeconomic characteristics and blood pressure among Jamaican youth: a pooled analysis of data from observational studies.
Neighbourhood characteristics are associated with several diseases, but few studies have investigated the association between neighbourhood and health in Jamaica. We evaluated the relationship between neighbourhood socioeconomic status (SES) and blood pressure (BP) among youth, 15-24 years old, in Jamaica. A pooled analysis was conducted using data from three studies (two national surveys and a birth cohort), conducted between 2005-2008, with individual level BP, anthropometric and demographic data, and household SES. Data on neighbourhood SES were obtained from the Mona Geo-Informatics Institute. Neighbourhood was defined using community boundaries from the Social Development Commission in Jamaica. Community characteristics (poverty, unemployment, dependency ratio, population density, house size, and proportion with tertiary education) were combined into SES scores using principal component analysis (PCA). Multivariable analyses were computed using mixed effects multilevel models. Analyses included 2,556 participants (1,446 females; 1,110 males; mean age 17.9 years) from 306 communities. PCA yielded two neighbourhood SES variables; the first, PCA-SES1, loaded highly positive for tertiary education and larger house size (higher value = higher SES); while the second, PCA-SES2, loaded highly positive for unemployment and population density (higher value = lower SES). Among males, PCA-SES1 was inversely associated with systolic BP (β-1.48 [95%CI -2.11, -0.84] mmHg, p < 0.001, for each standard deviation unit increase in PCA-SES1 score) in multivariable model accounting for age, household SES, study, BMI, fasting glucose, physical activity and diet. PCA-SES1 was not significantly associated with systolic BP among females (β -0.48 [-1.62, 0.66], p = 0.410) in a similar model. Associations for PCA-SES2 was assessed using linear splines to account for non-linear effects. The were no significant associations between systolic BP and PCA-SES2 among males. Among females, higher PCA-SES2 (i.e. lower SES) was associated with higher systolic BP at spline 2 [z-score -1 to 0] (β4.09 [1.49, 6.69], p = 0.002), but with lower systolic BP at spline 3 [z-core 0 to 1] (β-2.81 [-5.04, -0.59], p = 0.013). There were no significant associations between diastolic BP and PCA-SES1, but PCA-SES2 showed non-linear associations with diastolic BP particularly among males. Higher neighbourhood SES was inversely associated with systolic BP among male Jamaican youth; there were non-linear associations between neighbourhood SES and systolic BP among females and for diastolic BP for both males and females.


Undifferentiated, overlapping, and mixed connective tissue diseases.
Undifferentiated connective tissue disease (UCTD) is a term used by many rheumatologists to define a group of diffuse connective tissue disorders that lack definitive characteristics of any particular well-defined disorder. Overlapping connective tissue disease is often used interchangeably with UCTD but they both refer to diseases that are in evolution before all the characteristic clinical and laboratory symptoms are manifested. However, the clinical features of some of the overlapping connective tissue diseases appear to be better defined. The classical one is mixed connective tissue disease, where features of systemic lupus erythematosus, progressive systemic sclerosis, and polymyositis may exist together with a positive anti-extractable nuclear antibody and high titers of anti-ribonuclear protein antibody. This review attempts to clarify the confusion between these terms. The problems in the clinical and laboratory diagnosis of common connective tissue diseases that coexist are addressed and treatment options discussed. The long-term implications of making a diagnosis of a definitive connective tissue disease before all the required criteria are met should be kept in mind because the patient may never develop the disease and yet be subjected to psychological, social, and economic hardships.


The discovery and the development of bendamustine for the treatment of non-Hodgkin lymphoma.
Non-Hodgkin lymphoma (NHL) is a heterogeneous group of lymphoid malignancies. The treatment strategy for patients with NHL had remained unchanged until the advent of the era of molecular targeting agents. Although rituximab-containing chemotherapy has improved the response rates and survival of patients with B-cell NHL (B-NHL), several subtypes of B-NHL, especially indolent B-NHL and mantle cell lymphoma (MCL), remain incurable. Therefore, novel treatment modalities for B-NHL, especially for indolent B-NHL and MCL, are needed. Bendamustine is an old, but unique, multifunctional cytotoxic agent that exhibits structural similarity to alkylating agents and purine analogs. Areas covered: The basic aspects and preclinical development of bendamustine are summarized, followed by a discussion of the clinical development of bendamustine-based treatments for indolent B-NHL and MCL. Expert opinion: Bendamustine monotherapy or the use of bendamustine in combination with rituximab is highly effective against various types of hematological malignancies, especially relapsed or refractory indolent B-NHL, and exhibits an acceptable toxicity profile. Furthermore, bendamustine plus rituximab might be a promising treatment option for patients with untreated indolent B-NHL. Therefore, bendamustine has the potential to play an important role in the treatment of malignant lymphoma.


Electroosmotic Flow in Microchannels.
The electroosmotic flow induced by an applied electrostatic potential field through microchannels between two parallel plates and a 90 degrees bend is analyzed in this work. A nonlinear, two-dimensional Poisson-Boltzmann equation governing the electrical double-layer field and the Laplace equation governing the electrostatic field distribution in microchannels are numerically solved using a finite-difference method. A body force caused by the interaction between the electrical double-layer field and the applied electrostatic field is included in the full Navier-Stokes equations. The effects of the electrical double-layer field and the applied electrostatic field on the fluid velocity distribution, pressure drop, and skin friction are discussed. A small pressure drop along the parallel plates is detected, although it is always neglected in the literature. Pressure is not a constant across the channel height. The axial velocity profile is no longer flat across the channel height when the Reynolds number is large. A separation bubble is detected near the 90 degrees junction when the Reynolds number is large. Copyright 2001 Academic Press.


Genetic biomarkers in Brugada syndrome.
Brugada syndrome is an inherited arrhythmia syndrome predisposing to sudden cardiac death. Six years after its initial description as a clinical entity, the first mutations in SCN5A encoding the cardiac sodium channel Nav1.5 were reported. Over 300 mutations in SCN5A have since been described in addition to mutations in genes encoding Nav1.5 auxiliary units, potassium and calcium channels. This review summarizes the current knowledge on the genetics of Brugada syndrome, focusing on SCN5A, and discusses its use as a biomarker for diagnosis, prognosis and treatment.


Revolutionizing treatment outcomes in hepatitis C: managed care implications and considerations--a clinical overview.
Chronic hepatitis C virus (HCV) infection is highly prevalent and associated with a wide range of hepatic and extrahepatic complications. The treatment landscape for HCV infection has evolved to include regimens without interferon formulations and/or ribavirin. This change simplifies therapy, improves tolerability, and decreases therapy duration, while improving virologic response rates. This article, the first in a series of 3, will provide an overview of the hepatitis C disease state, identify populations at risk for HCV, describe testing recommendations for the diagnosis of HCV infection, and distinguish new and emerging HCV therapies.


Hypothyroidism and thyroiditis after therapy for Hodgkin's disease.
During the follow-up of thyroid function of 151 patients with Hodgkin's disease in complete remission for at least 1 year, 26 cases of subclinical, 12 cases of manifest clinical hypothyroidism and 2 cases of hyperthyroidism (Graves-Basedow disease) were confirmed. Thyroid dysfunction was more frequent in patients who had undergone mantle or neck radiotherapy. Hypothyroidism was most often revealed from the 6th year on following radiotherapy. Thyroid autoantibody positivity was found to be more frequent in patients with thyroid dysfunction, and conversely, thyroid dysfunction was more frequent among the 28 patients with autoantibody positivity. Ultrasound examination and fine needle aspiration cytology of the thyroid confirmed thyroiditis in 96% of the patients with antibody positivity. No relationship was found between thyroiditis and the form of treatment for Hodgkin's disease. We have found that both neck irradiation and thyroiditis may play a role in the increased number of thyroid dysfunction in patients treated for Hodgkin's disease. Thyroiditis is not caused by neck radiotherapy but may be the result of immune regulation disorders in Hodgkin's disease. For substitution or isohormone therapy, levothyroxine is suggested for use. We suggest that examination of the thyroid should be performed at least once a year during the follow-up of Hodgkin's disease patients.


Postmarket surveillance. Final rule.
The Food and Drug Administration (FDA) is implementing the postmarket surveillance (PS) provisions of the Federal Food, Drug, and Cosmetic Act (the act), as amended by the Food and Drug Administration Modernization Act of 1997 (FDAMA). The purpose of this rule is to provide for the collection of useful data about devices that can reveal unforeseen adverse events or other information necessary to protect the public health.


Expression of the human multidrug resistance and glucocerebrosidase cDNAs from adeno-associated vectors: efficient promoter activity of AAV sequences and in vivo delivery via liposomes.
Recombinant adeno-associated viruses (rAAV) are attractive tools for gene therapy. We designed plasmids in which the human multidrug resistance gene (hMDR1) cDNA was placed downstream from portions of the 5' end of AAV including either a 234-bp cassette or the entire AAV p5 promoter. The drug-resistant phenotype conferred by the P-glycoprotein (Pgp) efflux pump encoded by the hMDR1 cDNA was used to select NIH-3T3 cells transfected with these plasmids. The 234-bp region alone showed promoter activity similar in strength to that of the entire p5 promoter or the retroviral Harvey murine sarcoma virus long terminal repeat (LTR); this result demonstrates that the 234-bp cassette might be used as a small and efficient promoter in rAAV designed to express large genes approaching the packaging limit of AAV particles. After transfection of AAV-MDR1 vectors, the integration of MDR1 sequences into the host cell genome was demonstrated by fluorescent in situ hybridization (FISH). In addition, Southern analysis of low-molecular-weight DNA extracted from drug-resistant cells grown under continuous selection pressure indicated the persistence of nonintegrated AAV-MDR1 plasmids. Coordinate expression of Pgp and human glucocerebrosidase (hGC) was observed in drug-selected NIH-3T3 cells transfected with a bicistronic vector in which MDR1 cDNA was linked to hGC cDNA via the encephalomyocarditis internal ribosome entry site sequence. Moreover, following a single intravenous injection of the bicistronic vector complexed to cationic liposomes into recipient mice, delivery of MDR1 and GC cDNAs was achieved in all the organs we tested. Our results demonstrate that the efficiency of liposomes as vehicles for in vitro and in vivo gene delivery, the advantages of AAV-vectors, and the use of MDR1 as a selectable marker might be successfully combined in gene therapy protocols.


Synthesis and characterization of polymer nanocarriers for the targeted delivery of therapeutic enzymes.
Protein drugs, such as recombinant enzymes useful for detoxification and replacement therapies, have extraordinary specificity and potency. However, inherently inadequate delivery to target sites and rapid inactivation limit their medical utility. Using chaperone polymeric particles designed within an injectible size range (sub-micron) may help solve these shortcomings. Such nanocarriers would (i) prevent premature inactivation of encapsulated therapeutic protein cargoes, (ii) provide a carrier that can be surface decorated by targeting ligands, and (iii) optimize sub-cellular localization of the drug. This chapter describes the techniques successfully employed for the preparation of polymer nanocarriers (PNC) loaded with the antioxidant enzyme, catalase, and targeted to endothelial cells. Methods of PNC synthesis, loading with catalase, characterization, coupling of a targeting moiety, and in vitro testing of the enzymatic and targeting activities are provided here. Advantages and disadvantages of specific designs are discussed. Due to the modular nature of the targeting methodology employed, it is believed that these protocols will provide a solid foundation for the formulation of a wide variety of enzymatic drug targeting strategies.


Distance measures in terms of substitution processes.
Phylogenetic reconstruction from DNA or amino acid sequences relies heavily on suitable distance measures. A number of new distance measures (asynchronous, LogDet, and paralinear distances) which possess the desired property of tree additivity under fairly general models of sequence evolution have been proposed recently, but they are not well understood from a mechanistic point of view. We review them here in a unifying framework, which is the substitution process in continuous time. The emerging interpretation will also clarify the relationship among these distance measures. We also tackle situations with site-to-site variation of substitution rates which is well known to cause non-additive distances and inconsistent branch lengths. For homogeneous, stationary, time-reversible models, this may be repaired provided that the distribution of rates is known. In contrast, we will show that, for non-stationary models, different tree topologies may produce identical joint distributions of letters in pairs of sequences, given the same distribution of rates. This precludes the existence of any tree-additive pairwise distance measure.


Feasibility of resuming peritoneal dialysis after severe peritonitis and Tenckhoff catheter removal.
Published guidelines suggest that after an episode of severe peritonitis that requires Tenckhoff catheter removal, peritoneal dialysis can be resumed after a minimum of 3 wk. However, the feasibility of resuming peritoneal dialysis after Tenckhoff catheter removal remains unknown. One hundred patients were identified with peritonitis that did not respond to standard antibiotic therapy in a specific center. Their clinical course was reviewed; in all of them, Tenckhoff catheters were removed and reinsertion was attempted at least 4 wk later. In 51 patients, the Tenckhoff catheter was successfully reinserted and peritoneal dialysis was resumed (success group). In the other 49 patients, reinsertion failed and the patient was put on long-term hemodialysis (fail group). The patients were followed for 18.5 +/- 16.8 mo. The overall technique survival was 30.8% at 24 mo. In the success group, 11 patients were changed to long-term hemodialysis within 8 mo after their return to continuous ambulatory peritoneal dialysis. In the fail group, 18 of the 20 deaths occurred within 12 mo after conversion to long-term hemodialysis. After resuming peritoneal dialysis, there was a significant decline in net ultrafiltration volume (0.38 +/- 0.16 to 0.21 +/- 0.19 L; P = 0.03) and a trend of rise in dialysate-to-plasma ratios of creatinine at 4 h (0.664 +/- 0.095 to 0.725 +/- 0.095; P = 0.15). Forty-five patients (88.2%) required additional dialysis exchanges or hypertonic dialysate to compensate for the loss of solute clearance or ultrafiltration, although there was no significant change in dialysis adequacy or nutritional status. It was concluded that after an episode of severe peritonitis that required Tenckhoff catheter removal, only a small group of patients could return to peritoneal dialysis. An early assessment of peritoneal function after Tenckhoff catheter reinsertion may be valuable.


Analysis of X-chromosome inactivation in horse embryos.
To define the time of X-chromosome inactivation in the horse, 122 conceptuses were collected transcervically between Days 6 and 28 (ovulation = Day 0) and subjected to cytogenetic analysis: 59 of the embryos were divided and in 41 of these separate cytogenetic analysis of the embryonic disc and remaining tissues was possible. Conceptuses were measured and photographed before capsule removal, culture in the presence of 5-bromodeoxyuridine and subsequent fixation for cytogenetic analysis. On average, 15 slides were prepared per conceptus. C-banding was used to determine the sex of each conceptus and, in the females, the effects of 5-bromodeoxyuridine incorporation were revealed by u.v. irradiation which produced R-bands and allowed identification of the inactive X-chromosome. Inactivation of the X chromosome began gradually in the trophoblastic (+/- attached endoderm and mesoderm) cells around Day 7.5 and in the embryonic disc around Day 11.5. Cells with an inactive X-chromosome predominated in the trophoblast by Day 10.5 and in the disc by Day 12.5.


Warfarin therapy: a review of the literature since the Fifth American College of Chest Physicians' Consensus Conference on Antithrombotic Therapy.
Evidence-based medicine is currently a fashionable term. The evidence that warfarin is safe, effective, and cost beneficial in preventing stroke in AF, DVT treatment, and DVT prophylaxis is mounting. However, the evidence that warfarin remains underutilized in these conditions is also mounting. There is new evidence that supports conservative management of overanticoagulation without bleeding. The amount of time, if any, that patients are off warfarin for various procedures is being reduced. Warfarin interactions with other agents continue to be reported so that practitioners can avoid or treat them. Even the contraindication of warfarin in pregnancy is being reexamined. Those with expertise in anticoagulation therapy have an imperative to inform colleagues about the evidence in favor of warfarin.


Interaction of porcine circovirus type 2 replication with intracellular redox status in vitro.
Redox status influences replication of some viruses but its effect on porcine circovirus type 2 (PCV2), the primary causative agent of the emerging swine disease post-weaning multisystemic wasting syndrome is not known. The interaction of PCV2 replication with intracellular redox status in PK15 cells was examined in this study. Intracellular glutathione (GSH) was measured spectrophotometrically by reaction with 5, 5'-dithiobis (2-nitrobenzoic acid). Total superoxide dismutase activity (SOD) was assayed by inhibition of oxyamine oxidation by the xanthine oxidase system. Malondialdehyde (MDA) was assayed spectrophotometrically using the thiobarbituric acid reaction. Both quantification of PCV2 DNA by real-time polymerase chain reaction and indirect immunofluorescence of PCV2-infected cells were used to evaluate the replication of PCV2. Both GSH and SOD decreased significantly at 48 hours after PCV2 infection, whereas MDA concentration increased significantly after 48 hour post-infection. Furthermore, PCV2 replication in PK15 cells was significantly impaired after the elevation of intracellular GSH through treatment with the antioxidant N-acetyl-l-cysteine (NAC), a precursor in GSH synthesis. In contrast, PCV2 replication in PK15 cells was enhanced after reduction of GSH levels through H2O2-mediated oxidation. In addition, NAC treatment blocked the increase of virus replication induced by H2O2. This study suggests that PCV2 infection induces oxidative stress and that intracellular redox status influences PCV2 replication in PK15 cells.


Differences in bone and vitamin D metabolism between primary hyperparathyroidism and malignancy-associated hypercalcemia.
Bone and vitamin D metabolism are examined in patients with primary hyperparathyroidism (1 degree HPT), humoral hypercalcemia of malignancy (HHM), and local osteolytic hypercalcemia (LOH) with normal renal function. Among the bone resorption markers, T scores of total deoxypyridinoline (Dpyd) were highest in HHM and were significantly higher than those in 1 degree HPT. Among the formation markers, T scores of osteocalcin (OC) were highest in 1 degree HPT but were negative in HHM. The elevation in total Dpyd was associated with an increase in OC in 1 degree HPT, and the ratios of total Dpyd/OC were similar to those in controls. In contrast, many patients with HHM and LOH exhibited elevated total Dpyd and suppressed OC with increased total Dpyd/OC ratios, but the ratios varied widely. Serum 1,25-dihydroxyvitamin D [1,25(OH)2D] was elevated in 1 degrees HPT but was suppressed in HHM and LOH at any serum Ca levels. These results demonstrate that increased bone resorption is associated with enhanced bone formation in 1 degrees HPT but are uncoupled in many of the HHM and LOH patients, and that total Dpyd/OC ratio can be a useful index to estimate the coupling state of bone. It is suggested that the reduction in serum 1,25(OH)2D cannot be explained by an elevation in serum Ca in HHM and LOH, and that the differences in bone and vitamin D metabolism in HHM and LOH from those in 1 degree HPT may be caused by a common mechanism such as the secretion of some cytokines from tumors.


Impact of European settlement and land use changes on Great Barrier Reef river catchments reconstructed from long-term coral Ba/Ca records.
The increase in sediment and nutrient loads entering the coastal waters of the Great Barrier Reef (GBR) and the associated degradation of water quality represents a major threat to coral reefs. Although the strengthening of preventative management strategies remains a priority, there is a general lack of terrestrial runoff baseline information with respect to the spatial and temporal severity of disturbances associated with ongoing European-style land use practices. Here we use new and existing high-resolution coral Ba/Ca and luminescence records from the central Cairns region to the southern GBR shelf to reconstruct sediment fluxes discharged into the GBR from before European settlement in the 1860s to the present-day. Since the commencement of European settlement in the 1860s we document a tripling of flood-plume suspended sediment loads delivered by the Burdekin River to the GBR lagoon relative to 'natural' pre-European baseline levels. We show that this is indicative of a much more extreme degradation of the river catchments than hitherto appreciated with intensified discharge events particularly from the central and southern catchments carrying higher sediment loads. More-over from the 1930s onwards the Burdekin River, the largest source of both sediment and freshwater to the GBR, has also exhibited a progressive northwards expansion of its flood plume. This, together with increased variability of freshwater inputs indicated by coral luminescence records, now shows that the inner GBR not only continues to be impacted by increasing sediment/nutrient loads but is also subject to higher intensity river discharge events due to the loss of ground cover causing increased overland runoff and erosion.


Propagation of reticuloendotheliosis virus strain t in cell lines derived from avian lymphoid leukosis lymphoma and Marek's disease tumour.
Propagation of reticuloendotheliosis virus strain T (REV-T) was studied in avian lymphoid leukosis lymphoma-derived cell lines (LSCC-1104x5 and BK 3) and Marek's disease tumour-derived cell lines (MDCC-MSB1 and RP 1). These four cell lines showed cellular growth for a long time even after they were infected with REV-T. Production of non-oncogenic REV-T, replication non-defective virus, in these cell lines continued for 3 to 7 months. When culture fluids from 1104x5, MSB1 and RP 1, which were infected with oncogenic REV-T, were inoculated into 1-day-old chickens, acute development of lethal reticuloendotheliosis (RE) tumours occurred. Especially after the use of the fluids from 1104x5, propagation of oncogenic REV-T was observed for a long time, 7 months. Of the chickens inoculated with the culture fluid of any REV-T-infected cell lines, many chickens which survived for 4 weeks showed abnormal feathering, 'nakanuke'. The body weight of these chickens was significantly less than that of uninoculated chickens. No contamination with infectious REV was observed in the culture fluids from any uninfected line cells.


[Brain dysfunction in climacteric and prevention].
The risk of cardiovascular disease in women is relatively small before menopause but increases considerably afterwards. Postmenopausal women have on average a total cholesterol, LDL and VLDL cholesterol levels higher than premenopausal women; at the same time, HDL cholesterol serum level is lower. Postmenopausal estrogen replacement therapy (ERT) has been repeatedly proven to reduce the incidence of coronary heart disease as well as the mortality from cardiovascular disease. Furthermore, little is known about the potential risk of developing a stroke associated to hormonal changes involved in the menopause. Likewise the benefit of the ERT on reduction in stroke-legated mortality and morbidity is still debated. On the other hand, preliminary trials suggest a beneficial role of ERT in women with Alzheimer's disease (AD), probably due to the estrogen's cholinergic neurotrophic and neuroprotective effects. Moreover, recent reports suggest a possible additive beneficial effect of ERT to cholinergic treatments for AD In normal postmenopausal women the ERT not only improve cognitive and emotional functions but also increase verbal memory.


[Inertial processes in the auditory system during localization of moving sound sources].
The significant role of inertial features of the auditory system in localization of moving sound sources is presented. This inertia requires some time of observation about the movement in space of the auditory signals. Slower time of estimation of the localizing features of the moving sound source (as comparing with the unmoving one) is followed by the appearance of new important possibility of the auditory system, namely in perception and analysis of the parameters of moving sound sources. Meanwhile these inertial features are manifested by another significant change in the activity of the auditory system. After the cessation of the auditory signals we observed some aftereffects which manifested that the subjective perceived trajectory of the moving sound source is lengthening toward the direction of the existing movement even when the auditory signal is switched off. Stated above can serve as an evidence of presence in the auditory system some predictive possibilities concerning the direction of moving sound sources. This feature is essential from biological point of view for an adequate organisation of behaviour of the subjects.


Single nucleotide polymorphism discovery in barley using autoSNPdb.
Molecular markers are used to provide the link between genotype and phenotype, for the production of molecular genetic maps and to assess genetic diversity within and between related species. Single nucleotide polymorphisms (SNPs) are the most abundant molecular genetic marker. SNPs can be identified in silico, but care must be taken to ensure that the identified SNPs reflect true genetic variation and are not a result of errors associated with DNA sequencing. The SNP detection method autoSNP has been developed to identify SNPs from sequence data for any species. Confidence in the predicted SNPs is based on sequence redundancy, and haplotype co-segregation scores are calculated for a further independent measure of confidence. We have extended the autoSNP method to produce autoSNPdb, which integrates SNP and gene annotation information with a graphical viewer. We have applied this software to public barley expressed sequences, and the resulting database is available over the Internet. SNPs can be viewed and searched by sequence, functional annotation or predicted synteny with a reference genome, in this case rice. The correlation between SNPs and barley cultivar, expressed tissue type and development stage has been collated for ease of exploration. An average of one SNP per 240 bp was identified, with SNPs more prevalent in the 5' regions and simple sequence repeat (SSR) flanking sequences. Overall, autoSNPdb can provide a wealth of genetic polymorphism information for any species for which sequence data are available.


Tyrosine kinase receptor activation inhibits NPR-C in lung arterial smooth muscle cells.
We have previously demonstrated that expression of the atrial natriuretic peptide (ANP) clearance receptor (NPR-C) is reduced selectively in the lung of rats and mice exposed to hypoxia but not in pulmonary arterial smooth muscle cells (PASMCs) cultured under hypoxic conditions. The current study tested the hypothesis that hypoxia-responsive growth factors, fibroblast growth factors (FGF-1 and FGF-2) and platelet-derived growth factor-BB (PDGF-BB), that activate tyrosine kinase receptors can reduce expression of NPR-C in PASMCs independent of environmental oxygen tension. Growth-arrested rat PASMCs were incubated under hypoxic conditions (1% O2) for 24 h; with FGF-1, FGF-2, or PDGF-BB (0.1-20 ng/ml for 1-24 h); or with ANG II (1-100 nM), endothelin-1 (ET-1, 0.1 microM), ANP (0.1 microM), sodium nitroprusside (SNP, 0.1 microM), or 8-bromo-cGMP (0.1 mM) for 24 h under normoxic conditions. Steady-state NPR-C mRNA levels were assessed by Northern blot analysis. FGF-1, FGF-2, and PDGF-BB induced dose- and time-dependent reduction of NPR-C mRNA expression within 1 h at a threshold concentration of 1 ng/ml; hypoxia, ANG II, ET-1, ANP, SNP, or cGMP did not decrease NPR-C mRNA levels in PASMCs under the above conditions. Downregulation of NPR-C expression by FGF-1, FGF-2, and PDGF-BB was inhibited by the selective FGF-1 receptor tyrosine kinase inhibitor PD-166866 and mitogen-activated protein/extracellular signal-regulated kinase inhibitors U-0126 and PD-98059. These results indicate that activation of tyrosine kinase receptors by hypoxia-responsive growth factors, but neither hypoxia per se nor activation of G protein-coupled receptors, inhibits NPR-C gene expression in PASMCs. These results suggest that FGF-1, FGF-2, and PDGF-BB play a role in the signal transduction pathway linking hypoxia to altered NPR-C expression in lung.


Intergenerational concepts of adolescent sexuality: implications for community-based reproductive health care with Haitian immigrants.
Intergenerational conflict occurs when immigrant parents hold different values from those of their more rapidly acculturating offspring. These conflicts frequently involve disparate views related to sex roles and reproduction. A community-based study of 19 immigrant Haitian parent-adolescent pairs in South Florida compared their attitudes and values about sexuality and reproduction. Data were obtained through focused, open-ended interviews. Content analysis procedures at the level of words and phrases facilitated the categorization of responses. Data revealed considerable differences between parents and adolescents about the sources and types of information learned about reproduction and contraception, when such information is learned, and expectations regarding premarital sexual intercourse. Both parents and adolescents lacked accurate biomedical information about contraception, placed responsibility for contraceptive use primarily on the female partner, considered reproduction a natural rather than a medical event, and believed parents have the major responsibility for educating children about reproduction and contraception. Implications for culturally-appropriate health care center on increasing the role of the public health nurse in health education, minimizing intergenerational and intercultural conflict, and engaging the Haitian immigrant community in the promotion of reproductive health.


The evolving classification of soft tissue tumours - an update based on the new 2013 WHO classification.
The new World Health Organization (WHO) classification of soft tissue tumours was published in early 2013, almost 11 years after the previous edition. While the number of newly recognized entities included for the first time is fewer than that in 2002, there have instead been substantial steps forward in molecular genetic and cytogenetic characterization of this family of tumours, leading to more reproducible diagnosis, a more meaningful classification scheme and providing new insights regarding pathogenesis, which previously has been obscure in most of these lesions. This brief overview summarizes changes in the classification in each of the broad categories of soft tissue tumour (adipocytic, fibroblastic, etc.) and also provides a short summary of newer genetic data which have been incorporated in the WHO classification.


[The treatment of dentin hypersensitivity].
Dentine hypersensitivity is caused by a change in fluid flow in the dentinal tubules, which excites nerve endings located in the dentinal tubules and at the pulp-dentine border area. Traumatic oral hygiene procedures, excessive use of acid containing dietary fluids and certain dental treatments have been implicated in the occurrence of dentine hypersensitivity. Initial treatment relies on a positive doctor-patient relationship and the use of a desensitizing toothpaste containing strontium chloride, potassium nitrate or sodium citrate. If no relief is obtained, further treatment attempts need to be undertaken by the dentist--application of fluoride varnishes, potassium oxalate, resins or adhesives.


Acute disseminated encephalomyelitis (ADEM) after autologous peripheral blood stem cell transplant for non-Hodgkin's lymphoma.
Acute disseminated encephalomyelitis (ADEM) is a demyelinating disorder of the central nervous system with an acute clinical onset and a wide variability in severity and outcome. It usually follows a viral infection or an immunization and is thought to be immuno- mediated. We report a case of ADEM with a dramatic clinical onset in an autologous peripheral blood stem cell transplant (PBSCT) recipient for non-Hodgkin's lymphoma who developed the neurologic syndrome 12 days after PBSC reinfusion. This is the first report of ADEM in the setting of autologous PBSCT, a therapeutic procedure performed with increasing frequency in a wide variety of hematologic and solid malignancies.


Hydrogen plasma surface activation of silicon for biomedical applications.
Silicon has gradually been recognized to be an essential trace element in the normal metabolism of higher animals, and the role of silicon in the human body has aroused interests in the biomedical community. In fact, the interactions between silicon-based devices and the human body such as biosensors and microelectromechanical systems (MEMS) often suffer from poor biocompatibility. In this work, hydrogen plasma immersion ion implantation (H-PIII) is conducted to improve the bioactivity or bone conductivity of silicon. In order to investigate the formation mechanism of bone-like apatite on the surface of the hydrogen implanted silicon wafer, two comparative experiments, hydrogenation and argon bombardment, are performed. The H-PIII sample exhibits an amorphous surface consisting of Si-H bonds. After immersion in simulated body fluids, a negatively charged surface containing the functional group ([triple bond]Si-O-) is produced and bone-like apatite is observed to nucleate and grow on the surface. The surface of the H-PIII silicon wafer favors the adhesion and growth of osteoblast cells and good cytocompatibility may be inferred.


Inhibition of type 1 and type 2 5alpha-reductase activity by free fatty acids, active ingredients of Permixon.
In different cell systems, the lipido-sterolic extract of Serenoa repens (LSESr, Permixon inhibits both type 1 and type 2 5alpha-reductase activity (5alphaR1 and 5alphaR2). LSESr is mainly constituted of fatty acids (90+/-5%) essentially as free fatty acids (80%). Among these free fatty acids, the main components are oleic and lauric acids which represent 65% and linoleic and myristic acids 15%. To evaluate the inhibitory effect of the different components of LSESr on 5alphaR1 or 5alphaR2 activity, the corresponding type 1 and type 2 human genes have been cloned and expressed in the baculovirus-directed insect cell expression system Sf9. The cells were incubated at pH 5.5 (5alphaR2) and pH 7.4 (5alphaR1) with 1 or 3nM testosterone in presence or absence of various concentrations of LSESr or of its different components. Dihydrotestosterone formation was measured with an automatic system combining HPLC and an on-line radiodetector. The inhibition of 5alphaR1 and 5alphaR2 activity was only observed with free fatty acids: esterified fatty acids, alcohols as well as sterols assayed were inactive. A specificity of the fatty acids in 5alphaR1 or 5alphaR2 inhibition has been found. Long unsaturated chains (oleic and linolenic) were active (IC(50)=4+/-2 and 13+/-3 microg/ml, respectively) on 5alphaR1 but to a much lesser extent (IC(50)>100 and 35+/-21 microg/ml, respectively) on 5alphaR2. Palmitic and stearic acids were inactive on the two isoforms. Lauric acid was active on 5alphaR1 (IC(50)=17+/-3 microg/ml) and 5alphaR2 (IC(50)=19+/-9 microg/ml). The inhibitory activity of myristic acid was evaluated on 5alphaR2 only and found active on this isoform (IC(50)=4+/-2 microg/ml). The dual inhibitory activity of LSESr on 5alpha-reductase type 1 and type 2 can be attributed to its high content in free fatty acids.


1,3-substituted imidazolidine-2,4,5-triones: synthesis and inhibition of cholinergic enzymes.
A series of novel and highly active acetylcholinesterase and butyrylcholinesterase inhibitors derived from substituted benzothiazoles containing an imidazolidine-2,4,5-trione moiety were synthesized and characterized. The molecular structure of 1-(2,6-diisopropyl-phenyl)-3-[(1R)-1-(6-fluoro-1,3-benzothiazol-2-yl)ethyl]-imidazolidine-2,4,5-trione (3g) was determined by single-crystal X-ray diffraction. Both optical isomers are present as two independent molecules in the triclinic crystal system. The lipophilicity of the compounds was determined as the partition coefficient log K(ow) using the traditional shake-flask method. The in vitro inhibitory activity on acetylcholinesterase from electric eel and butyrylcholinesterase isolated from equine serum was determined. The inhibitory activity on acetylcholinesterase was significantly higher than that of the standard drug rivastigmine. The discussed compounds are also promising inhibitors of butyrylcholinesterase, as some of the prepared compounds inhibit butyrylcholinesterase better than the internal standards rivastigmine and galanthamine. The highest inhibitory activity (IC₅₀ = 1.66 μmol/L) corresponds to the compound 1-(4-isopropylphenyl)-3-[(R)-1-(6-fluorobenzo[d]thiazol-2-yl)ethyl]imidazolidine-2,4,5-trione (3d). For all the studied compounds, the relationships between the lipophilicity and the chemical structure as well as their structure-activity relationships are discussed.


Integration sites and genotype distributions of human papillomavirus in cervical intraepithelial neoplasia.
To analyse HPV integration prevalence and genotype distributions in cervical intraepithelial neoplasia (CIN) in east part of China, furthermore to assess preferential sites for common HPV integrations and provide baseline information for cervical abnormality screening and prevention. Integration of HPV in 113 paraffin-embedded cervical intraepithelial neoplasia samples was assessed using Gencap technology in Key Laboratory of Biotechnologies in BGI-Shenzhen. 64 samples were HPV-integrated and as the cervical lesions increased, the integration rate became higher significantly (P=0.002). Fifteen different HPV genotypes were detected, 14 high-risk (16, 18, 31, 33, 51, 52, 56, 58, 66, 68) and 1 low-risk (11). The most common genotypes were HPV-16, 58, 33, 52, 66, and 56. Thirteen patients had co-integration involving mainly HPV-16 and 58. The frequency of HPV gene disruption was higher in L1 and E1 genes than in other regions of the viral genomes. Some 56.6% of CIN lesions in Qingdao had HPV integrations, and 67.2% of HPV-integrated patients were HPV-16 and 58, more prone to be integrated in younger patients below 45 years old. There exist preferential sites for HPV-16 and HPV-58 integration, and they are more likely to be disrupted in the L1 and E1 loci.


The microtubule binding properties of CENP-E's C-terminus and CENP-F.
CENP-E (centromere protein E) and CENP-F (centromere protein F), also known as mitosin, are large, multi-functional proteins associated with the outer kinetochore. CENP-E features a well-characterized kinesin motor domain at its N-terminus and a second microtubule-binding domain at its C-terminus of unknown function. CENP-F is important for the formation of proper kinetochore-microtubule attachment and, similar to CENP-E, contains two microtubule-binding domains at its termini. While the importance of these proteins is known, the details of their interactions with microtubules have not yet been investigated. We have biochemically and structurally characterized the microtubule-binding properties of the amino- and carboxyl-terminal domains of CENP-F as well as the carboxyl-terminal (non-kinesin) domain of CENP-E. CENP-E's C-terminus and CENP-F's N-terminus bind microtubules with similar affinity to the well-characterized Ndc80 complex, while CENP-F's C-terminus shows much lower affinity. Electron microscopy analysis reveals that all of these domains engage the microtubule surface in a disordered manner, suggesting that these factors have no favored binding geometry and may allow for initial side-on attachments early in mitosis.


Selective Delivery of an Anticancer Drug with Aptamer-Functionalized Liposomes to Breast Cancer Cells in Vitro and in Vivo.
Selective targeting of cancer cells is a critical step in cancer diagnosis and therapy. To address this need, DNA aptamers have attracted significant attention as possible targeting ligands. However, while their use in targeting cancer cells in vitro has been reported, their effectiveness has rarely been established in vivo. Here we report the development of a liposomal drug delivery system for targeted anticancer chemotherapy. Liposomes were prepared containing doxorubicin as a payload, and functionalized with AS1411, a DNA aptamer with strong binding affinity for nucleolin. AS1411 aptamer-functionalized liposomes increased cellular internalization and cytotoxicity to MCF-7 breast cancer cells as compared to non-targeting liposomes. Furthermore, targeted liposomal doxorubicin improved antitumor efficacy against xenograft MCF-7 breast tumors in athymic nude mice, attributable to their enhanced tumor tissue penetration. This study suggests that AS1411 aptamer-functionalized liposomes can recognize nucleolin overexpressed on MCF-7 cell surface, and therefore enable drug delivery with high specificity and selectivity.


Microsphere Synthesis by Dispersion Copolymerizations Using Poly(t-butyl methacrylate) Macromonomers in Nonaqueous Media.
Polymer microspheres were synthesized by dispersion copolymerization of divinylbenzene (DVB) with vinylbenzyl-terminated poly(t-butyl methacrylate) (PBMA) macromonomers in nonaqueous media (methanol and acetonitrile). The particle diameter increased gradually with an increase of the DVB concentration in methanol or acetonitrile. The particle size distribution was very narrow. PBMA macromonomers acted not only as a comonomer but also as a stabilizer. The particle diameters obtained in acetonitrile were smaller than those obtained in methanol. Thus, we observed a tendency in which the more soluble for PBMA macromonomer the media became, the smaller the particle size became. Copyright 2001 Academic Press.


Symposium: Update on prediction and management of OHSS. Prevention of OHSS--dopamine agonists.
Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic complication of ovulation induction, which may seriously affect the patient's health, with 0.1-2.0% of the patients developing severe forms of the syndrome. OHSS signs and symptoms are direct consequences of fluid shift from capillaries to the third space due to increased vascular permeability. The mechanisms of OHSS pathophysiology remain unknown but evidence obtained in animal models prove that vascular endothelial growth factor (VEGF) is directly involved and the inhibition of the VEGF system could prevent OHSS from occurring. Dopamine agonists impede the phosphorylation of the VEGF receptor 2 and reduce the incidence and severity of OHSS with a safe clinical profile. As far as is known, this is the first pathophysiological approach for treatment and prevention of OHSS.


RBCK1 drives breast cancer cell proliferation by promoting transcription of estrogen receptor alpha and cyclin B1.
Cell cycle regulatory pathways in breast cancer are incompletely described. Here, we report an important role in estrogen receptor alpha (ERalpha)-positive breast cancer cells for the protein kinase C1 (PKC1)-interacting protein RBCK1 in supporting cell cycle progression by driving transcription of ERalpha and cyclin B1. RBCK1-depleted cells exhibited increased accumulation in G(2)-M phase of the cell cycle, decreased proliferation, and reduced mRNA levels for ERalpha and its target genes cyclin D1 and c-myc. Chromatin immunoprecipitation revealed that ERalpha transcription is associated with RBCK1 recruitment to the ERalpha promoter, suggesting that transcriptional regulation is one mechanism by which RBCK1 affects ERalpha mRNA levels. G(2)-M phase arrest was mediated independently from reduced ERalpha levels, instead associated with transcriptional inhibition of the key G(2)-M regulator cyclin B1. In breast tumor samples, there was a positive correlation between levels of RBCK1, ERalpha, and cyclin B1 mRNA levels. Our findings suggest that RBCK1 regulates cell cycle progression and proliferation of ERalpha-positive breast cancer cells by supporting transcription of ERalpha and cyclin B1.


Stereoisomer quantification of the beta-blocker drugs atenolol, metoprolol, and propranolol in wastewaters by chiral high-performance liquid chromatography-tandem mass spectrometry.
A chiral liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) method was developed and validated for measuring individual enantiomers of three beta-blocker drugs (atenolol, metoprolol, and propranolol) in wastewater treatment plant (WWTP) influents and effluents. Mean recoveries of the pharmaceuticals ranged from 67 to 106%, and the limits of detection of the analytes were 2-17 ng/L in wastewater effluents. The method was demonstrated by measuring, for the first time, the stereoisomer composition of target analytes in raw and treated wastewaters of two Canadian WWTPs. In these trials, racemic amounts of the three drugs were observed in influent of one wastewater treatment plant, but nonracemic amounts were observed in another. Effluents of the two plants contained nonracemic amounts of the drugs. These results indicate that biologically-mediated stereoselective processes that differ among WWTPs had occurred to eliminate individual enantiomers of the target analytes.


A review on biofiltration techniques: recent advancements in the removal of volatile organic compounds and heavy metals in the treatment of polluted water.
Good quality of water determines the healthy life of living beings on this earth. The cleanliness of water was interrupted by the pollutants emerging out of several human activities. Industrialization, urbanization, heavy population, and improper disposal of wastes are found to be the major reasons for the contamination of water. Globally, the inclusion of volatile organic compounds (VOCs) and heavy metals released by manufacturing industries, pharmaceuticals, and petrochemical processes have created environmental issues. The toxic nature of these pollutants has led researchers, scientists, and industries to exhibit concern toward the complete eradication of them. In this scenario, the development of wastewater treatment methodologies at low cost and in an eco-friendly way had gained importance at the international level. Recently, bio-based technologies were considered for environmental remedies. Biofiltration-based works have shown a significant result for the removal of volatile organic compounds and heavy metals in the treatment of wastewater. This was done with several biological sources such as bacteria, fungi, algae, plants, yeasts, etc. The biofiltration technique is cost-effective, simple, biocompatible, sustainable, and eco-friendly compared to conventional techniques. This review article provides deep insight into biofiltration technologies engaged in the removal of volatile organic compounds and heavy metals in the wastewater treatment process.


Better communication between experts is needed to solve the environmental origins of birth defects.
More than 6% of babies are born with a structural or functional defect, and many of these need special care and treatment to survive and thrive. Such defects can be inherited, arise through exposure to altered conditions or compounds in the womb, or result from a combination of genetic and environmental factors. Since the 1940s, animal experiments and epidemiological studies have identified many environmental factors that can cause particular birth defects. More recently, advances in genomics have allowed a simple genetic diagnosis in ∼ 30% of birth defects. However, the cause of the remainder is a mystery. I believe that a key limiter to successful identification of new environmental factors is that clinicians, epidemiologists and developmental biologists all approach the topic from different angles. I propose that better communication between such experts will further increase our understanding of the environmental causes of birth defects, and potentially reduce their global burden.


Active and inactive renin in the cat.
Pentobarbitone-anesthetized cats underwent peritoneal dialysis and had blood samples removed and kidneys deep frozen at sacrifice. Inactive renin is easily measurable in cat plasma and peritoneal dialysate fluid. Only small amounts are found after acid activation at pH 4.0, but large amounts after trypsin 2 mg/ml at 4 degrees C for 10 minutes. Mean active renin in pentobarbitone-anesthetized cats was 1.8 +/- 0.4 pmoles AI/ml/hr, while inactive renin was 2.3 +/- 0.5 pmoles AI/ml/hr. The increased angiotensin I producing activity after trypsin in peritoneal dialysate was most active at pH 7.0 (plasma and kidney active renin 7.25 and 7.85), and had an apparent molecular weight of 39-40,000. (Plasma active renin had an apparent MW of 33,500 and kidney active renin 36,000. Plasma inactive renin had an apparent MW of 35,500) Cat plasma after cibacron-blue affinity chromatography showed mainly active renin in the breakthrough buffer (30% of total renin eluted), and renin which is almost entirely inactive in the bound peak (70% of total renin eluted). Active renin from plasma and kidney, and activated inactive renin from concentrated peritoneal fluid, showed exactly similar inhibition by the renin inhibitor H77 (IC50 0.3 microM). Cat plasma angiotensinogen had an apparent MW of 53,000.


Detection of Border disease virus in fetuses, stillbirths, and newborn lambs from natural and experimental infections.
The purpose of the present study was to evaluate the use of enzyme-linked immunosorbent assay (ELISA) antigen detection in blood or fetal fluids and reverse transcription polymerase chain reaction (RT-PCR) amplification in tissues for routine laboratory diagnosis of Border disease virus (BDV) infection. Samples from 67 fetuses, 6 stillbirths, and 11 lambs from 25 commercial flocks with suspicion of BDV abortion and 3 fetuses, 7 stillbirths, and 15 lambs obtained from an experimental infection with a local isolate (BDV genotype 4) were investigated. Presence of BDV was detected by RT-PCR in 7.9% of fetuses, 50% of stillbirths, and 50% of lambs from the commercial flocks analyzed, corresponding to 8 of the 25 farms (32%). A similar percentage of the lambs and stillbirths from the experimental infection were positive by RT-PCR of tissue samples (54.5%), and the highest positivity was detected in lymph node, thyroid gland, and kidney. The current study revealed that RT-PCR analysis of stillbirths and lambs with clinical symptoms is more suitable than the analysis of fetuses to confirm the presence of BDV in a flock. Pestiviral antigen was detected by antigen ELISA in a high proportion of fetuses (24/58) and stillbirths (3/4) from commercial flocks, but in lambs, the presence of colostral antibodies masked the detection of the antigen by ELISA. Nevertheless, in lambs from the experimental infection that were not fed colostrum, antigen ELISA was less efficient than RT-PCR in detecting viral presence in stillbirths and lambs. Antigen ELISA is therefore recommended for fetuses with advanced autolysis that can adversely affect RNA integrity.