Pharmacodynamics in the development of new immunosuppressive drugs.
Over the past 10-20 years a number of immunosuppressive drugs, such as cyclosporine A, tacrolimus, sirolimus, or mycophenolate mofetil have been approved for clinical use and have been highly successful in preventing or delaying graft rejection. Nevertheless, there is an incessant need for better and safer drugs to improve short-term and long-term outcomes following transplantation. A number of low-molecular-weight molecules that interfere with immune cell functions are in development. These include molecules that inhibit the janus protein tyrosine kinase JAK3, compounds that alter lymphocyte trafficking (the sphingosine-1-phosphate receptor antagonist FTY720), and new malononitrilamides (FK778). All seem to show promising therapeutic potential. Among the biologic agents, there are high expectations for antibodies or recombinant chimeric molecules targeting costimulatory surface molecules or pathways involved in the migration of immune cells. The list of such targets includes the ligand pairs CD28:B7, CD154:CD40, LFA-1:ICAM-1, ICOS:B7RP-1, and VLA-4:VCAM-1. However, the clinical development of drugs for transplantation has proved to be difficult, complex, and time consuming. Therefore, newly emerging drug candidates will also demand better methods for monitoring their efficacy as well as their side effects in vivo. Pharmacokinetics (PK) and pharmacodynamics (PD) are complementary approaches used to select drugs on the basis of their in vivo efficacy as well as safety. Whereas PK monitors the handling of the drug by the body, PD focuses on the biologic effect of the drug on its target. Therefore, PD studies of in vivo efficacy are useful for clinical decisions to determine the optimal dose and type of immunosuppressant. At the preclinical stage, PD is aimed at accelerating the selection of lead compounds via PD-controlled trials in animals. Moreover, PD can help to discover new mechanisms of action for a drug or a drug candidate. However, its full potential has not been used, mainly because of laborious and time-consuming methodology. This review focuses on established and novel PD/PK approaches to assess immunosuppressive compounds in the context of new evolving drugs or drug combinations.


Diurnal hormone variation in fibromyalgia syndrome: a comparison with rheumatoid arthritis.
Twenty patients with fibromyalgia syndrome and 20 patients with rheumatoid arthritis (RA) were assessed as outpatients over a 3 day period with respect to peak and trough levels of plasma cortisol, growth hormone, prolactin, ACTH and thyroid stimulating hormone. Patients with fibromyalgia syndrome had loss of diurnal variation in plasma cortisol (trough levels 347.3 +/- 254.7 vs 232.8 +/- 70.0 nmol/l, p less than 0.001) compared with RA patients. Thirty-five percent (7/20) of patients with fibromyalgia syndrome and only 5 percent (1/20) of those with RA exhibited abnormal dexamethasone suppression tests (p less than 0.001). No differences were noted in the diurnal variation of other hormones tested. Beck Depression Inventory scores were similar in both groups and no patient exhibited clinical evidence of depression. These data suggest alteration in the pituitary hypothalamic axis with respect to cortisol secretion in fibromyalgia syndrome, perhaps as a consequence of chronic pain.


Mass scaling of metabolic rates in carabid beetles (Carabidae) - the importance of phylogeny, regression models and gas exchange patterns.
The origin of the allometric relationship between standard metabolic rate (MR) and body mass (M), often described as MR=aMb , remains puzzling, and interpretation of the mass-scaling exponent, b may depend on the methodological approach, shapes of residuals, coefficient of determination (r2) and sample size. We investigated the mass scaling of MRs within and between species of Carabidae beetles. We used ordinary least squares (OLS) regression, phylogenetically generalized least squares (PGLS) regression and standardized major axis (SMA) regression to explore the effects of different model-fitting methods and data clustering caused by phylogenetic clades (grade shift) and gas exchange patterns (discontinuous, cyclic and continuous). At the interspecific level, the relationship between MR and M was either negatively allometric (b<1) or isometric (b=1), depending on the fitting method. At the intraspecific level, the relationship either did not exist or was isometric or positively allometric (b>1), and the fit was significantly improved after the analyzed dataset was split according to gas exchange patterns. The studied species originated from two distinct phylogenetic clades that had different intercepts but a common scaling exponent (OLS, 0.61) that was much shallower than the scaling exponent for the combined dataset for all species (OLS, 0.71). The best scaling exponent estimates were obtained by applying OLS while accounting for grade shifts or by applying PGLS. Overall, we show that allometry of MR in insects can depend heavily on the model fitting method, the structure of phylogenetic non-independence and ecological factors that elicit different modes of gas exchange.


Adhesion complex formation after small keratectomy wounds in the cornea.
The adhesion complex of the corneal epithelium consists of the hemidesmosome and its associated structures, such as anchoring filaments, lamina densa of the basement membrane, and anchoring fibrils. It contributes to the adhesion of the corneal epithelium to Bowman's layer. To understand the adhesion complex better, an electron microscopic and immunofluorescence analysis was done of the reformation of the adhesion complex in small (1 mm) keratectomy wounds in the guinea pig cornea. In these wounds, the epithelium, hemidesmosomes, basal lamina, anchoring fibrils, and anterior stroma were removed. The wound bed was epithelialized completely by 24 hr after wounding. Immunofluorescence analyses involved the use of antibodies against plaque components of the hemidesmosome, an antibody against laminin, and an antibody against the collagen VII component of anchoring fibrils. At 18 hr after wounding, there was no morphologic evidence of hemidesmosomes at the epithelial-stromal interface. At 24 hr, hemidesmosomes were observed, with or without subjacent lamina densa. Furthermore, plaque components were detected by immunofluorescence in those cells in contact with the wound bed. In contrast, no type VII collagen was detected. On day 7, collagen VII, laminin, and bullous pemphigoid autoantibody markers colocalized along the wound bed as determined by immunofluorescence. However, at the ultrastructural level, even though the lamina densa of the basal lamina was observed primarily where hemidesmosomes were present, it remained incomplete. In this study, the precise temporal sequence in which components are incorporated into the assembling adhesion complex was described during wound healing. Furthermore, the possibility that the hemidesmosomal plaque nucleates the formation of the underlying basal lamina was discussed.


FSRD: fungal stress response database.
Adaptation to different types of environmental stress is a common part of life for today's fungi. A deeper understanding of the organization, regulation and evolution of fungal stress response systems may lead to the development of novel antifungal drugs and technologies or the engineering of industrial strains with elevated stress tolerance. Here we present the Fungal Stress Response Database (http://internal.med.unideb.hu/fsrd) aimed to stimulate further research on stress biology of fungi. The database incorporates 1985 fungal stress response proteins with verified physiological function(s) and their orthologs identified and annotated in 28 species including human and plant pathogens, as well as important industrial fungi. The database will be extended continuously to cover other fully sequenced fungal species. Our database, as a starting point for future stress research, facilitates the analysis of literature data on stress and the identification of ortholog groups of stress response proteins in newly sequenced fungal genomes. Database URL: http://internal.med.unideb.hu/fsrd


Possible direct influence of complement 3 in decreasing insulin sensitvity in a cohort of overweight and obese subjects.
A group of 608 apparently healthy patients, 136 men and 472 women, either overweight or obese, aged 18-69 years, were examined. BMI, waist circumference, fasting blood glucose (FBG), insulin, and complement 3 (C3) serum levels were measured; the homeostasis model assessment (HOMAIR) was used to evaluate insulin resistance; and physical activity was quantified by a questionnaire. HOMAIR showed a positive correlation with BMI (r: 0.478, p < 0.001), waist circumference (r: 0.487, p < 0.001), and C3 (r: 0.445, p < 0.001). Moreover, it was significantly associated with gender (F Fisher = 22.12, p < 0.001), and the mean HOMAIR levels were significantly different among the three groups of physical activity, with the lowest level of insulin resistance at the highest level of physical activity (F=7,31, p < 0.001). A multiple regression analysis was carried out with HOMAIR as the dependent variable and gender, age, BMI, waist circumference, C3 and the level of physical activity as independent variables (fitted model: F = 41.24, P<0.001, R2 = 0.328). HOMAIR maintained an independent association with C3 (β = 0.678, P<0.001), sex (β = 0.189, P<0.001), BMI (β = 0.637, P<0.01), and age (β = -0.004, P<0.05). This study of a cohort of overweight and obese subjects has shown that insulin resistance (dependent variable) is positively associated with C3 serum levels, independently of age, gender, anthropometric parameters and physical activity, suggesting that higher C3 serum levels may directly increase insulin resistance in obesity.


Rail-roading technique using 18 gauge intravenous catheter and silicon rod for frontalis suspension in blepharophimosis syndrome.
Silicon rods are widely employed for frontalis sling suspension. However, on passing through the tissues, at times, the silicon rod gets detached from the stainless steel needle. This occurs more commonly in patients of blepharophimosis syndrome, in which hypoplasia of superior orbital rim with deficiency of skin between lid and brow, causes difficulty in passage of the needle when it is manipulated upwards from lid towards the brow. To overcome these problems we describe the use of an 18 G intravenous catheter to railroad the needle with the silicon rod, obviating the blind upward maneuvers with the needle and protecting against the damage to the silicon rod -needle assembly. The technique is easily reproducible, safe and can be used in all silicon rod suspensions.


Homologous DNA exchanges in humans can be explained by the yeast double-strand break repair model: a study of 17p11.2 rearrangements associated with CMT1A and HNPP.
Rearrangements in 17p11.2, responsible for the 1.5 Mb duplications and deletions associated, respectively, with autosomal dominant Charcot-Marie-Tooth type 1A disease (CMT1A) and hereditary neuropathy with liability to pressure palsies (HNPP) are a suitable model for studying human recombination. Rearrangements in 17p11.2 are caused by unequal crossing-over between two homologous 24 kb sequences, the CMT1A-REPs, that flank the disease locus and occur in most cases within a 1.7 kb hotspot. We sequenced this hotspot in 28 de novo patients (25 CMT1A and three HNPP), in order to localize precisely, at the DNA sequence level, the crossing-overs. We show that some chimeric CMT1A-REPs in de novo patients (10/28) present conversion of DNA segments associated with the crossing-over. These rearrangements can be explained by the double-strand break (DSB) repair model described in yeast. Fine mapping of the de novo rearrangements provided evidence that the successive steps of this model, heteroduplex DNA formation, mismatch correction and gene conversion, occurred in patients. Furthermore, the model explains 17p11.2 recombinations between chromosome homologues as well as between sister chromatids. In addition, defective mismatch repair of the heteroduplex DNA, observed in two patients, resulted in two heterozygous chimeric CMT1A-REPs which can be explained, as in yeast, by post-meiotic segregation. This work supports the hypothesis that the DSB repair model of DNA exchange may apply universally from yeasts to humans.


Solubilization of the ileal receptor for intrinsic factor--vitamin B-12 complex by digestion with papain.
Brush-border-membrane vesicles isolated from hamster ileum were incubated with either papain or Pronase P and subsequently centrifuged to obtain soluble (supernatant) and insoluble (pellet) fractions. Papain (4 units/ml) solubilized 95--100% of the sucrase and leucine naphthylamide-hydrolysing activities but only 30% of the alkaline phosphatase. Digestion with papain also resulted in the solubilization of more than 75% of the ileal receptor for intrinsic factor-vitamin B-12 complex with a corresponding decrease in receptor activity in the pellet. Essentially 100% of the receptor activity was recovered. In contrast, digestion with Pronase P resulted in a decrease in total receptor activity. Papain-solubilized receptor was not sedimented by centrifugation at 105 000 g for 90 min and was eluted in the included volume of Sepharose 6B. Like the binding to more intact preparations, binding of intrinsic factor-vitamin B-12 complex to papain-solubilized receptor was rapid, reaching 50% of maximum in 8 min, and required Ca2+. Although Mg2+ could not completely substitute for Ca2+, Mg2+ did stimulate Ca2+-dependent binding at low Ca2+ concentrations. These results demonstrate that the ileal receptor for intrinsic factor-vitamin B-12 complex can be solubilized with papain, and suggest that papain solubilization may be a useful first step in the isolation and purification of this receptor.


Specific localization of a plant cell wall glycine-rich protein in protoxylem cells of the vascular system.
An antibody against glycine-rich protein 1.8 of bean (Phaseolus vulgaris L.) was used for immunogold/silver localization of the protein in different organs of the plant. In hypocotyls, ovaries, and seed coats, the protein was found specifically in xylem cells of the vascular tissue. In hypocotyls, only protoxylem cells were labeled with the antibody, which indicates a remarkable cell-type specificity for accumulation of this cell wall protein. In mature hypocotyls, the protein was restricted to the same subset of xylem cells but was no longer detected on tissue prints, where a positive antibody reaction depends on the transfer of soluble material from plant tissue to the nitrocellulose filter. This indicates that the glycine-rich protein is insolubilized in the cell wall during development. In longitudinal sections of tracheary elements of young hypocotyls and seed coats, the antibody stained a pattern very similar to that of the lignified secondary thickenings of the cell wall, which suggests a close functional relationship between glycine-rich protein and lignin deposition during cell wall biogenesis in protoxylem cells.


Outcome of per protocol best-evidence based routine breast cancer care in a large regional hospital in Belgium: the importance of a prospective database in quality assurance.
Criteria for future accreditation of breast cancer centres in Belgium will be mainly based on the case load per surgeon or per centre. We would like to argue that the prospective collection of relevant data and the analysis of treatment related outcome derived from these data is feasible and should be the ultimate criterion for quality assessment and thus for accreditation since outcome is a more direct measurement of quality. Data were prospectively collected on 715 invasive non metastatic breast cancers between 2002 and 2007 treated according to standard, best-evidence protocols in the setting of a large district hospital. Univariate and multivariate survival analysis were performed and compared to national and international databases. 5 year disease-free survival (DFS) and overall survival (OS) in our series were respectively 77 and 84%. In the multivariate analysis of DFS, only her-2-neu status (her-2-neu positivity being associated with a poor prognosis) and age (older age being a worse prognostic factor) were statistically significant prognostic factors. For OS, her-2-neu, age, and positive nodes were statistically significant prognostic factors. The outcome is comparable to other data sets. Centres dedicated to the care of women with breast cancer have the moral duty to produce outcome based results of their treatment. This report shows that such a collection of data is feasible and can be imposed as a prerequisite for accreditation. We also argue that outcome based data of treatment are a more solid base for quality assurance than case load.


Congenital deficiency of vitamin K-dependent coagulation factors and protein C.
A 15-month-old girl from Coimbra (Portugal) had a history of numerous hemorrhagic episodes with multiple bruises, hematomas but not hemarthroses. On serial testing she showed deficiency of factors II, VII, IX, X and protein C. Malabsorption-induced vitamin K deficiency, liver disease or ingestion of a coumarin compound were excluded. An absence of detectable abnormalities was found among her relatives. Consanguinity was not present. The immunologic assay, immunoelectrophoresis or antibody neutralization, revealed much higher levels of these factors than the clotting assay. The non-physiological activator (Echis carinatus venom) produced higher levels of prothrombin activation than those detected by physiological activation. Two-dimensional immunoelectrophoresis of the patient's plasma in calcium showed that prothrombin had the same mobility as acarboxyprothrombin. No significant response to large doses of intravenous vitamin K3 (6 mg) was observed. Transfusion of 120 ml of frozen fresh plasma led to an immediate increase in the procoagulant activities of vitamin K dependent protein, similar to that found after perfusion of plasma plus vitamin K3. The results obtained from this patient suggest a defect in the gammacarboxylation mechanism inside the hepatocyte.


Congress of Neurological Surgeons Systematic Review and Evidence-Based Guideline for the Management of Patients With Residual or Recurrent Nonfunctioning Pituitary Adenomas.
Despite the advancement of microsurgical and endoscopic techniques, some nonfunctioning pituitary adenomas (NFPAs) can be difficult to cure. Tumor recurrence or incomplete resection may occur in some patients with NFPAs, and management strategies for these NFPAs remain unclear. To review the existing literature as it pertains to the management of postsurgical residual or recurrent NFPAs. A systematic review of the treatment options for residual or recurrent NFPAs was performed. In this review, the authors critically evaluated the evidence to support the options of repeat microsurgical resection, stereotactic radiosurgery (SRS), stereotactic radiotherapy (SRT), and fractionated radiation therapy (XRT). Forty-nine studies met the inclusion criteria for analysis: outcome of repeat surgical resection (n = 4), radiosurgery (ie, single-session or hypofractionated SRS; n = 24), or fractionated radiotherapy (ie, conventional XRT, proton beam radiotherapy, intensity-modulated radiotherapy, SRT; n = 21). No class I evidence was available; 6 studies met criteria for class II evidence; and other studies provided class III evidence. A meta-analysis of 5 class II studies with recurrence rates for both adjuvant radiation therapy and observation demonstrated that XRT for residual/recurrent NFPAs offered a lower rate of recurrence (odds ratio = 0.04; 95% confidence interval, 0.01-0.20; P < .001). The analysis also demonstrated significant heterogeneity between the included studies (χ = 20.70; P = .003; I = 81%). Repeat resection, SRS, SRT, and XRT play a role in the management of patients with recurrent or residual NFPAs. SRS or some type of radiation therapy is typically performed for patients with residual tumor or tumor recurrence after resection. The full guidelines document for this chapter can be located at https://www.cns.org/guidelines/guidelines-management-patients-non-functioning-pituitary-adenomas/Chapter_7. NFPA, nonfunctioning pituitary adenomaSRS, stereotactic radiosurgerySRT, stereotactic radiotherapyXRT, fractionated radiation therapy.


Protective cytotoxic T lymphocyte responses induced by DNA immunization against immunodominant and subdominant epitopes of Listeria monocytogenes are noncompetitive.
Taking advantage of the fact that plasmid DNA encoding a single cytotoxic T lymphocyte (CTL) epitope can induce CTLs, we examined the influence of T-cell responses to dominant epitopes on those to a subdominant epitope derived from Listeria monocytogenes. Our data suggest that interaction between T cells against dominant and subdominant epitopes does not operate in the generation of the hierarchy. Furthermore, we found that a single dominant epitope is sufficient for the induction of protective immunity.


Readability of Wikipedia Pages on Autoimmune Disorders: Systematic Quantitative Assessment.
In the era of new information and communication technologies, the Internet is being increasingly accessed for health-related information. Indeed, recently published patient surveys of people with autoimmune disorders confirmed that the Internet was reported as one of the most important health information sources. Wikipedia, a free online encyclopedia launched in 2001, is generally one of the most visited websites worldwide and is often consulted for health-related information. The main objective of this investigation was to quantitatively assess whether the Wikipedia pages related to autoimmune disorders can be easily accessed by patients and their families, in terms of readability. We obtained and downloaded a list of autoimmune disorders from the American Autoimmune Related Diseases Association (AARDA) website. We analyzed Wikipedia articles for their overall level of readability with 6 different quantitative readability scales: (1) the Flesch Reading Ease, (2) the Gunning Fog Index, (3) the Coleman-Liau Index, (4) the Flesch-Kincaid Grade Level, (5) the Automated Readability Index (ARI), and (6) the Simple Measure of Gobbledygook (SMOG). Further, we investigated the correlation between readability and clinical, pathological, and epidemiological parameters. Moreover, each Wikipedia analysis was assessed according to its content, breaking down the readability indices by main topic of each part (namely, pathogenesis, treatment, diagnosis, and prognosis plus a section containing paragraphs not falling into any of the previous categories). We retrieved 134 diseases from the AARDA website. The Flesch Reading Ease yielded a mean score of 24.34 (SD 10.73), indicating that the sites were very difficult to read and best understood by university graduates, while mean Gunning Fog Index and ARI scores were 16.87 (SD 2.03) and 14.06 (SD 2.12), respectively. The Coleman-Liau Index and the Flesch-Kincaid Grade Level yielded mean scores of 14.48 (SD 1.57) and 14.86 (1.95), respectively, while the mean SMOG score was 15.38 (SD 1.37). All the readability indices confirmed that the sites were suitable for a university graduate reading level. We found no correlation between readability and clinical, pathological, and epidemiological parameters. Differences among the different sections of the Wikipedia pages were statistically significant. Wikipedia pages related to autoimmune disorders are characterized by a low level of readability. The onus is, therefore, on physicians and health authorities to improve the health literacy skills of patients and their families and to create, together with patients themselves, disease-specific readable sites, disseminating highly accessible health-related online information, in terms of both clarity and conciseness.


Adult Coping with Childhood Sexual Abuse: A Theoretical and Empirical Review.
Coping has been suggested as an important element in understanding the long-term functioning of individuals with a history of child sexual abuse (CSA). The present review synthesizes the literature on coping with CSA, first by examining theories of coping with trauma, and, second by examining how these theories have been applied to studies of coping in samples of CSA victims. Thirty-nine studies were reviewed, including eleven descriptive studies of the coping strategies employed by individuals with a history of CSA, eighteen correlational studies of the relationship between coping strategies and long-term functioning of CSA victims, and ten investigations in which coping was examined as a mediational factor in relation to long-term outcomes. These studies provide initial information regarding early sexual abuse and subsequent coping processes. However, this literature is limited by several theoretical and methodological issues, including a failure to specify the process of coping as it occurs, a disparity between theory and research, and limited applicability to clinical practice. Future directions of research are discussed and include the need to understand coping as a process, identification of coping in relation to adaptive outcomes, and considerations of more complex mediational and moderational processes in the study of coping with CSA.


Use of the UFOV to evaluate and retrain visual attention skills in clients with stroke: a pilot study.
The objective of this pilot study was to examine the use of a visual attention analyzer in the evaluation and retraining of useful field of view in clients with stroke. Fifty-two clients with stroke referred to a driving evaluation service were evaluated with a visual attention analyzer referred to as the UFOV. The UFOV assesses three aspects of visual attention: processing speed, divided attention, and selective attention. Seven participants were retested to determine the test-retest reliability of the UFOV. Six participated in the development of a training protocol and in a 20-session visual attention retraining program. UFOV scores indicated substantial reduction in visual attention in clients after stroke, with older participants performing the most poorly. Test-retest reliability was moderate (ICC = .70). Mean UFOV scores improved significantly after retraining. Although UFOV scores indicated poor visual attention skills in clients with stroke, preliminary information suggests that UFOV scores significantly improve with training.


Luminol and gold nanoparticle-co-precipitated reduced graphene oxide hybrids with long-persistent chemiluminescence for cholesterol detection.
Herein, dual-functionalized reduced graphene oxide (rGO) hybrids (rGO/AuNP/luminol) with gold nanoparticles (AuNPs) and luminol were prepared viaπ-π conjugation and electrostatic interaction. The as-prepared rGO/AuNP/luminol hybrids exhibited enhanced and long-persistent chemiluminescence (CL) performance without decay for one hour when mixed with H2O2; this was due to the generation of ˙OH, O2˙-, 1O2, and π-C[double bond, length as m-dash]C˙ species. A sensitive and selective CL sensor based on the rGO/AuNP/luminol hybrid CL enhancing system was developed for cholesterol quantitation in the linear range from 0.71 to 11.43 μM with a detection limit of 0.55 μM. Furthermore, the proposed method was successfully exploited for the determination of cholesterol in human serum samples.


Gestational and neonatal iron deficiency alters apical dendrite structure of CA1 pyramidal neurons in adult rat hippocampus.
The hippocampus develops rapidly during the late fetal and early postnatal periods. Fetal/neonatal iron deficiency anemia (IDA) alters the genomic expression, neurometabolism and electrophysiology of the hippocampus during the period of IDA and, strikingly, in adulthood despite neonatal iron treatment. To determine how early IDA affects the structural development of the apical dendrite arbor in hippocampal area CA1 in the offspring, pregnant rat dams were given an iron-deficient (ID) diet between gestational day 2 and postnatal day (P) 7 followed by rescue with an iron-sufficient (IS) diet. Apical dendrite morphology in hippocampus area CA1 was assessed at P15, P30 and P70 by Scholl analysis of Golgi-Cox-stained neurons. Messenger RNA levels of nine cytoplasmic and transmembrane proteins that are critical for dendrite growth were analyzed at P7, P15, P30 and P65 by quantitative real-time polymerase chain reaction. The ID group had reduced transcript levels of proteins that modify actin and tubulin dynamics [e.g. cofilin-1 (Cfl-1), profilin-1 (Pfn-1), and profilin-2 (Pfn-2)] at P7, followed at P15 by a proximal shift in peak branching, thinner third-generation dendritic branches and smaller-diameter spine heads. At P30, iron treatment since P7 resulted in recovery of all transcripts and structural components except for a continued proximal shift in peak branching. Nevertheless, at P65-P70, the formerly ID group showed a 32% reduction in 9 mRNA transcripts, including Cfl-1 and Pfn-1 and Pfn-2, accompanied by 25% fewer branches, that were also proximally shifted. These alterations may be due to early-life programming of genes important for structural plasticity during adulthood and may contribute to the abnormal long-term electrophysiology and recognition memory behavior that follows early iron deficiency.


Exposure of the US population to environmental tobacco smoke: the Third National Health and Nutrition Examination Survey, 1988 to 1991.
To estimate the extent of exposure of the US population to environmental tobacco smoke and the contribution of the home and workplace environment to environmental tobacco smoke exposure. Nationally representative cross-sectional survey including questionnaire information from persons aged 2 months and older (n=16818) and measurements of serum cotinine (a metabolite of nicotine) from persons aged 4 years and older (n=10642). Participants in the Third National Health and Nutrition Examination Survey, October 25, 1988, to October 21, 1991. Of US children aged 2 months to 11 years, 43% lived in a home with at least 1 smoker, and 37% of adult non-tobacco users lived in a home with at least 1 smoker or reported environmental tobacco smoke exposure at work. Serum cotinine levels indicated more widespread exposure to nictoine. Of non-tobacco users, 87.9% had detectable levels of serum cotinine. Both the number of smokers in the household and the hours exposed at work were significantly and independently associated (P<.001, multiple regression t test) with increased serum cotinine levels. Serum cotinine levels of children, non-Hispanic blacks, and males indicated that these groups had higher exposure to environmental tobacco smoke. Dietary variables showed no consistent association with serum cotinine levels, and dietary contribution to serum cotinine level, if any, appeared to be extremely small. The high proportion of the population with detectable serum cotinine levels indicates widespread exposure to environmental tobacco smoke in the US population. Both the home and workplace environments significantly contribute to environmental tobacco smoke exposure in the United States.


Nonsurgical endodontic treatment of dens invaginatus in a mandibular premolar with large periradicular lesion: a case report.
This case report describes the treatment of a dens invaginatus in a mandibular premolar with large periradicular lesion. A nonsurgical endodontic treatment was performed to tooth #28. During the treatment procedure, calcium hydroxide and a triple antibiotic paste were separately used for intracanal dressing. The apical third of the canal was obturated with mineral trioxide aggregate (MTA), and the remaining canal space was obturated with gutta-percha and AH 26 root canal sealer, using a lateral condensation technique. At follow-up examination after 1 year, the tooth was asymptomatic and radiographically showed repair of the lesion. Healing was achieved without any need for further endodontic surgical intervention.


Silver enhancement of polymerised diaminobenzidine: increased sensitivity for immunoperoxidase staining.
Unambiguous identification of lymphocytes is sometimes difficult because of weak immunostaining of the cell membrane immunoglobulins. A simple method of intensifying the diaminobenzidine (DAB) peroxidase reaction was therefore devised. Paraffin wax sections of formalin fixed tonsils and lymphomas were digested with trypsin and immunostained for kappa and lambda light immunoglobulin chains and CD3 antigen by various peroxidase linked detection systems. After reaction with hydrogen peroxide and DAB the sections were immersed in methenamine silver solution at 60 degrees C for three to seven minutes. The light brown stain on the cell membranes of the mantle zone lymphocytes became dark brown and the stronger stain of the plasma cells became black. Mantle zone B lymphocytes and CD3 positive T lymphocytes were precisely outlined even at low magnification and the lymphomas were easily classified as monoclonal or polyclonal. At high magnification, staining was clearer than with the immunogold-silver stain. Cryostat and paraffin wax sections of other tissues immunostained for various antigens showed similar intensification. Silver methenamine provides an easy means of increasing the sensitivity and visual impact of an immunoperoxidase/DAB reaction in any preparation.


Clinical utility of reliable digit span in assessing effort in children and adolescents with epilepsy.
The assessment of effort is an important aspect of a comprehensive neuropsychological evaluation, as this can significantly impact data interpretation. While recent work has validated the appropriateness of adult-derived cutoffs for standalone effort measures in younger populations, little research has focused on embedded effort measures in children. The present study includes 54 clinically referred children and adolescents (32 males/22 females; aged 6-17) with a confirmed diagnosis of epilepsy. Reliable Digit Spans (RDSs) were calculated and the Test of Memory Malingering (TOMM) was administered in the context of a comprehensive neuropsychological evaluation. Using a previously published RDS cutoff of ≤6, a pass rate of only 65% was obtained, well below the recommended 90% pass rate for an effective effort index. In contrast, when adult criteria were used on TOMM Trial 2, a 90% pass rate was observed. RDS scores were significantly correlated with IQ estimates (r = .59, p < .001) and age (r = .61, p < .001). The difference between RDS and the TOMM on the participant outcome was statistically significant (χ(2) = 9.05, p = .003). These results suggest that RDS appears to yield a large number of false positives and, therefore, may be of limited utility in detecting poor effort in a pediatric epilepsy population. These findings likely extend to other pediatric populations that are known to have significant cognitive loss.


New fundamental resistance exercise determinants of molecular and cellular muscle adaptations.
Abstract Physical activity relies on muscular force. In adult skeletal muscle, force results from the contraction of postmitotic, multinucleated myofibres of different contractile and metabolic properties. Myofibres can adapt to (patho-)physiological conditions of altered functional demand by radial growth, longitudinal growth, and regulation of fibre type functional gene modules. The adaptation's specificity depends on the distinct molecular and cellular events triggered by unique combinations of conditional cues. In order to derive effective and tailored exercise prescriptions, it must be determined (1) which mechano-biological condition leads to what molecular/cellular response, and (2) how this molecular/cellular response relates to the structural, contractile, and metabolic adaptation. It follows that a thorough mechano-biological description of the loading condition is imperative. Unfortunately, the definition of (resistance) exercise conditions in the past and present literature is insufficient. It is classically limited to load magnitude, number of repetitions and sets, rest in-between sets, number of interventions/week, and training period. In this review, we show why the current description is insufficient, and identify new determinants of quantitative and/or qualitative effects on skeletal muscle with respect to resistance exercise in healthy, adult humans. These new mandatory determinants comprise the fractional and temporal distribution of the contraction modes per repetition, duration of one repetition, rest in-between repetitions, time under tension, muscular failure, range of motion, recovery time, and anatomical definition. We strongly recommend to standardise the design and description of all future resistance exercise investigations by using the herein proposed set of 13 mechano-biological determinants (classical and new ones).


TreeKnit: Inferring ancestral reassortment graphs of influenza viruses.
When two influenza viruses co-infect the same cell, they can exchange genome segments in a process known as reassortment. Reassortment is an important source of genetic diversity and is known to have been involved in the emergence of most pandemic influenza strains. However, because of the difficulty in identifying reassortment events from viral sequence data, little is known about their role in the evolution of the seasonal influenza viruses. Here we introduce TreeKnit, a method that infers ancestral reassortment graphs (ARG) from two segment trees. It is based on topological differences between trees, and proceeds in a greedy fashion by finding regions that are compatible in the two trees. Using simulated genealogies with reassortments, we show that TreeKnit performs well in a wide range of settings and that it is as accurate as a more principled bayesian method, while being orders of magnitude faster. Finally, we show that it is possible to use the inferred ARG to better resolve segment trees and to construct more informative visualizations of reassortments.


Validation of Human Sterol 14α-Demethylase (CYP51) Druggability: Structure-Guided Design, Synthesis, and Evaluation of Stoichiometric, Functionally Irreversible Inhibitors.
Sterol 14α-demethylases (CYP51) are the cytochrome P450 enzymes required for biosynthesis of sterols in eukaryotes, the major targets for antifungal agents and prospective targets for treatment of protozoan infections. Human CYP51 could be and, for a while, was considered as a potential target for cholesterol-lowering drugs (the role that is now played by statins, which are also in clinical trials for cancer) but revealed high intrinsic resistance to inhibition. While microbial CYP51 enzymes are often inhibited stoichiometrically and functionally irreversibly, no strong inhibitors have been identified for human CYP51. In this study, we used comparative structure/functional analysis of CYP51 orthologs from different biological kingdoms and employed site-directed mutagenesis to elucidate the molecular basis for the resistance of the human enzyme to inhibition and also designed, synthesized, and characterized new compounds. Two of them inhibit human CYP51 functionally irreversibly with their potency approaching the potencies of azole drugs currently used to inhibit microbial CYP51.


Antireflux Surgery Versus Antireflux Medication and Risk of Esophageal Adenocarcinoma in Patients With Barrett's Esophagus.
Antireflux treatment is recommended to reduce esophageal adenocarcinoma in patients with Barrett's esophagus. Antireflux surgery (fundoplication) counteracts gastroesophageal reflux of all types of carcinogenic gastric content and reduces esophageal acid exposure to a greater extent than antireflux medication (eg, proton pump inhibitors). We examined the hypothesis that antireflux surgery prevents esophageal adenocarcinoma to a larger degree than antireflux medication in patients with Barrett's esophagus. This multinational and population-based cohort study included all patients with a diagnosis of Barrett's esophagus in any of the national patient registries in Denmark (2012-2020), Finland (1987-1996 and 2010-2020), Norway (2008-2020), or Sweden (2006-2020). Patients who underwent antireflux surgery were compared with nonoperated patients using antireflux medication. The risk of esophageal adenocarcinoma was calculated using multivariable Cox regression, providing hazard ratios (HRs) and 95% CIs adjusted for age, sex, country, calendar year, and comorbidity. The cohort consisted of 33,939 patients with Barrett's esophagus. Of these, 542 (1.6%) had undergone antireflux surgery. During up to 32 years of follow-up, the overall HR was not decreased in patients having undergone antireflux surgery compared with nonoperated patients using antireflux medication, but rather increased (adjusted HR, 1.9; 95% CI, 1.1-3.5). In addition, HRs did not decrease with longer follow-up, but instead increased for each follow-up category, from 1.8 (95% CI, 0.6-5.0) within 1-4 years of follow-up to 4.4 (95% CI, 1.4-13.5) after 10-32 years of follow-up. Patients with Barrett's esophagus who undergo antireflux surgery do not seem to have a lower risk of esophageal adenocarcinoma than those using antireflux medication.


Efficacy of Vonoprazan for Gastroesophageal Reflux Symptoms in Patients with Proton Pump Inhibitor-resistant Non-erosive Reflux Disease.
Objective Clinically, patients with proton pomp inhibitor (PPI)-resistant gastroesophageal reflux disease (GERD) are very challenging to treat. The aim of this study was to determine the rates of symptom relief and adverse events among PPI-resistant GERD patients that changed their therapy from a PPI to vonoprazan. Methods Patients with severe gastroesophageal reflux symptoms (total GERD-Q score ≥8) without endoscopic findings of mucosal breaks who changed their medication from a PPI to vonoprazan during a 12-week period from 2015 to 2016 at 2 hospitals were selected. The primary outcome was the self-reported relief of gastroesophageal reflux symptoms. The odds ratio (OR) for the improvement of symptoms was calculated based on an exact binomial distribution using a matched-pair analysis. The secondary outcome was the GERD-Q score and adverse events. Results Twenty-six patients (6 men) with a mean age of 67.5 years were analyzed. After the therapy was changed from a PPI to vonoprazan, 18 patients (69.2%) reported an improvement, 6 (23.1%) reported no change, and 2 (7.7%) reported an exacerbation of symptoms. A change in therapy was significantly associated with improved self-reported symptoms (OR 9.0, p<0.001). The mean total GERD-Q score during vonoprazan treatment was significantly lower than that during PPI therapy (11.96 vs. 8.92). There were no significant differences in the incidence of adverse events between the therapies. Conclusion Changing the medication from a PPI to vonoprazan was significantly associated with an improvement in gastroesophageal reflux symptoms. Vonoprazan is one of the most promising treatment options for patients with PPI-resistant GERD.


The validity of three-class Hotelling trace (3-HT) in describing three-class task performance: comparison of three-class volume under ROC surface (VUS) and 3-HT.
In order to describe multiclass classification performance, several figures of merit (FOM) have been proposed. Among the earliest and most widely known of these is the three-class Hotelling trace (3-HT). The goal of this paper is to present theoretical and empirical data demonstrating the failure of 3-HT as a measure of three-class task performance. To help do this, we contrast it to a newly proposed three-class FOM, the volume under the three-class receiver operating characteristic (ROC) surface (VUS). The VUS is obtained from a decision theory based three-class ROC analysis method which has been proved to extend the decision theoretic, linear discriminant analysis (LDA), and psychophysical foundations of binary ROC analysis to a three-class paradigm. We demonstrate empirically that the VUS and 3-HT do not have a monotonic relationship in general when describing three-class task performance. Numerical experiments demonstrated that the VUS provided reasonable results, while the 3-HT failed to distinguish between the case where all objects could be perfectly classified from the case where only one pair of the classes could be perfectly classified. We have provided theoretical explanations of this failure of 3-HT. The significance of this work goes beyond merely demonstrating the problems of the 3-HT, it demonstrates that a FOM that is mathematically correct and has a strong theoretical basis can provide results that violate a common sense understanding of three-class task performance. This fact raises the question of "how to evaluate a classification performance evaluation method?" We believe the answer to this question lies in the theoretical foundations of binary ROC analysis. We have thus contrasted the two FOMs in terms of three fundamental theories underlying binary ROC analysis: decision theory, binary linear discriminant analysis, and the equivalence of two psychophysical classification procedures. These theoretical investigations demonstrated the importance of extending and unifying all the fundamental theories of binary classification in the development of a three-class FOM; violating one of theses fundamental binary classification theories may, as it did for the L-HT, provide predictions of three-class task performance that do not agree with a common sense understanding of three-class task performance.


High levels of the circulating form of parathyroid hormone do not inhibit in vitro erythropoiesis.
Either primary or secondary hyperparathyroidism may be associated with anemia. The pathogenesis of this anemia remains obscure, but parathyroid hormone may directly suppress erythropoiesis or anemia may result from myelofibrosis. Previously reported in vitro studies of a direct inhibitory effect of PTH on erythroid progenitor growth were carried out with crude hormone preparations and appeared nonspecific. We have studied simultaneously the in vitro effects of pure (greater than 6000 U/mg) and crude (130 U/mg) preparations of PTH on murine and human hematopoietic progenitor growth. Increasing concentrations (2.5 to 20 U/ml) of crude PTH produced a dose-dependent inhibition of early erythroid (BFU-E) and granulocyte/macrophage progenitor (CFU-GM) growth in human and mouse marrow cell cultures. However, the biologically active N-terminal fragment containing amino acids 1-34 and the pure intact molecule of 84 amino acids failed to significantly inhibit hematopoietic colony growth. These observations demonstrate that in vitro inhibitory effects described with parathyroid gland extracts are not specific for erythropoiesis and may not be related to the circulating form of PTH.


Sleep habits and road traffic accident risk for Iranian occupational drivers.
The aim of this study is to assess the sleep quality and sleep disorders (prevalence of obstructive sleep apnea and sleepiness) among occupational drivers in Iran and to determine which demographic factors and occupational habits are linked to road traffic accidents. In this analytic cross-sectional study 556 occupational road drivers from Shahroud city (in the northeast of Iran) participated, upon a prior verbal informed consent, during 2013-2014. The Pittsburgh Sleep Quality Index (PSQI) standard questionnaire that scored on 7 point scale, the 8-item Epworth Sleepiness Scale (ESS) questionnaire and the 8-question STOP-Bang questionnaire along with demographic information and occupational data were used. To explore the independent factors associated with odds of poor sleep quality and road accident, multiple logistic regression models were used. Prevalence of previous road accidents, sleepiness while driving, and obstructive sleep apnea scored ≥ 3 in the study, and drivers accounted for 23.8%, 29%, and 24.8%, respectively. The global mean score of sleep quality and excessive sleepiness score were 5.2 and 4.8, respectively. The main factors related to the odds of poor sleep quality were snoring (odds ratio (OR) = 2.34; 95% confidence interval (CI): 1.15-4.77), smoking (OR = 2.12; 95% CI: 1.15-3.97), and driving times in a day (OR = 1.12; 95% CI: 1.03-1.21). The Epworth Sleepiness Scale (OR = 1.13; 95% CI: 1.07-1.23) and suffering from apnea (OR = 4.89; 95% CI: 1.07-23.83) were the best predictors for odds (increased risk) of road accidents. A considerable proportion of Iranian drivers had records of road accidents; poor sleep quality, sleepiness while driving, and sleep disorder breathing (obstructive sleep apnea - OSA). Snoring, smoking, driving time in a day, excessive sleepiness, and presumably apnea increase the odds of poor sleep quality and road traffic accident for Iranian occupational drivers.


Lens orientalis Contributes Quantitative Trait Loci and Candidate Genes Associated With Ascochyta Blight Resistance in Lentil.
Australian lentil production is affected by several major biotic constraints including Ascochyta blight (AB), caused by Ascochyta lentis, a devastating fungal disease. Cultivation of AB resistant cultivars, alongside agronomic management including fungicide application, is the current most economically viable control strategy. However, the breakdown of AB resistance in cultivars, such as Northfield and Nipper, suggests the need for introgression of new and diverse resistance genes. Successful introgression entails an understanding of the genetic basis of resistance. In this context, a biparental mapping population derived from a cross between a recently identified AB resistant accession ILWL 180 (Lens orientalis) and a susceptible cultivar ILL 6002 was produced. A genetic linkage map was constructed from single-nucleotide polymorphism markers generated using a genotyping-by-sequencing transcript approach. Genetic dissection of the mapping population revealed a major quantitative trait loci (QTL) region nested with three QTLs on linkage group 5 and explained 9.5-11.5 percent (%) of phenotypic variance for AB resistance. Another QTL was identified on LG2 with phenotypic variance of 9.6%. The identified QTL regions harbored putative candidate genes potentially associated with defense responses to A. lentis infection. The QTL analysis and the candidate gene information are expected to contribute to the development of diagnostic markers and enable marker-assisted resistance selection in lentil breeding programmes.


Pure red cell aplasia and hypogammaglobulinemia associated with Tr-cell chronic lymphocytic leukemia.
A 72-yr-old male with Tr-cell chronic lymphocytic leukemia (Tr-CLL) exhibited pure red cell aplasia (PRCA) and hypogammaglobulinemia. During a remission of Tr-CLL, and while receiving cyclophosphamide therapy, he recovered from PRCA and hypogammaglobulinemia. To investigate the pathogenesis of PRCA and hypogammaglobulinemia, we used coculture techniques to study the effect of the malignant Tr cells on erythroid colony formation and B-cell differentiation to immunoglobulin-producing cells. Varying numbers of malignant Tr cells (2 X 10 to 2 X 10(5) cells) were cocultured with 2 X 10(5) normal bone marrow cells. The malignant Tr cells caused a marked reduction of erythroid colony formation in the plasma clot system. This suppression of erythroid colony formation was reversed when the malignant Tr cells were pretreated with antilymphocyte serum and complement. There was no evidence of inhibitory effects in the serum or the supernatant media of the malignant Tr cells stimulated with phytohemagglutinin (PHA). The malignant Tr cells, stored at --80 degrees C before transfusion, were also capable of suppressing autologous erythroid colony formation after recovery from PRCA. In addition, malignant Tr cells were found to have strong suppressor activity against the immunoglobulin biosynthesis by allogeneic B cells. The in vitro suppressions of both erythroid colony formation and B-cell differentiation provide an explanation for the association of PRCA and hypogammaglobulinemia with Tr-CLL.


Liquid-stored red blood cells for transfusion. A status report.
Blood transfusion in a modern sense means the transfusion of red cells, when necessary supplemented by other components. The demand for plasma and plasma fractions and for platelets for therapeutic use has had an influence on the technique for preparing red cells. Automated devices have made it possible to perform collection as well as separation under more standardized conditions. Improved techniques for storage of red cells have prolonged the shelf life somewhat but most of the available methods disregard the rapid loss of 2,3-diphosphoglycerate and the accompanying increase in oxygen affinity. Methods are available which reduce the number of contaminating leukocytes to low levels, but information is still incomplete as to the degree of depletion actually needed.


Enhanced active extracellular polysaccharide production from Ganoderma formosanum using computational modeling.
Extracellular polysaccharide (EPS) is one of the major bioactive ingredients contributing to the health benefits of Ganoderma spp. In this study, response surface methodology was applied to determine the optimal culture conditions for EPS production of Ganoderma formosanum. The optimum medium composition was found to be at initial pH 5.3, 49.2 g/L of glucose, and 4.9 g/L of yeast extract by implementing a three-factor-three-level Box-Behnken design. Under this condition, the predicted yield of EPS was up to 830.2 mg/L, which was 1.4-fold higher than the one from basic medium (604.5 mg/L). Furthermore, validating the experimental value of EPS production depicted a high correlation (100.4%) with the computational prediction response model. In addition, the percentage of β-glucan, a well-recognized bioactive polysaccharide, in EPS was 53±5.5%, which was higher than that from Ganoderma lucidum in a previous study. Moreover, results of monosaccharide composition analysis indicated that glucose was the major component of G. formosanum EPS, supporting a high β-glucan percentage in EPS. Taken together, this is the first study to investigate the influence of medium composition for G. formosanum EPS production as well as its β-glucan composition.


Cyp2c44 epoxygenase is essential for preventing the renal sodium absorption during increasing dietary potassium intake.
The aim of this study is to test whether the Cyp2c44 epoxygenase-dependent metabolism of arachidonic acid prevents the hypertensive effect of a high K (HK) intake by inhibiting the epithelial sodium channel (ENaC) activity. A HK intake elevated Cyp2c44 mRNA expression and 11,12-epoxyeicosatrienoic acid levels in the cortical collecting duct in Cyp2c44(+/+) mice (wild-type [wt]). However, an HK intake failed to increase 11,12-epoxyeicosatrienoic acid formation in the cortical collecting ducts of Cyp2c44(-/-) mice. Moreover, increasing K intake enhanced arachidonic acid-induced inhibition of ENaC in the wt but not in Cyp2c44(-/-) mice. In contrast, 11,12-epoxyeicosatrienoic acid, a Cyp2c44 metabolite, inhibited ENaC in the wt and Cyp2c44(-/-) mice. The notion that Cyp2c44 is the epoxygenase responsible for mediating the inhibitory effects of arachidonic acid on ENaC is further suggested by the observation that inhibiting Cyp-epoxygenase increased the whole-cell Na currents in principal cells of wt but not in Cyp2c44(-/-) mice. Feeding mice with an HK diet raised the systemic blood pressures of Cyp2c44(-/-) mice but was without an effect on wt mice. Moreover, application of amiloride abolished the HK-induced hypertension in Cyp2c44(-/-) mice. The HK-induced hypertension of Cyp2c44(-/-) mice was accompanied by decreasing 24-hour urinary Na excretion and increasing the plasma Na concentration, and the effects were absent in wt mice. In contrast, disruption of the Cyp2c44 gene did not alter K excretion. We conclude that Cyp2c44 epoxygenase mediates the inhibitory effect of arachidonic acid on ENaC and that Cyp2c44 functions as an HK-inducible antihypertensive enzyme responsible for inhibiting ENaC activity and Na absorption in the aldosterone-sensitive distal nephron.


Results of treatment of arteriovenous fistulae with the Debrun technique.
In a series of 54 caroticocavernous fistulae and eight vertebrovertebral fistulae, clinical cures were obtained in 46 of the caroticocavernous fistulae and in all of the vertebrovertebral fistulae using the Debrun coaxial catheter detachable balloon technique. Most low vertebrovertebral and about half of the posteriorly opening caroticocavernous fistulae were successfully treated by the safe transvenous approach. Technical difficulties were not infrequent but were overcome in most cases. In nine cases the artery was occluded with the fistula and in two others minimal residual shunt persisted, progressing in one to a symptomatic aneurysm. Five caroticocavernous fistulae in which the balloon could not be satisfactorily sited were not closed, and another caroticocavernous fistula closed spontaneously after a failed transvenous approach. Displacement of balloons during detachment reopened two caroticocavernous fistulae. One of these had to be closed by transcranial surgery; the other balloon embolized the middle cerebral artery, was surgically removed, and a mild hemiparesis persisted. Premature balloon detachment in a grossly atheromatous artery caused the only other major complication and contributed to the death of an 87-year-old patient. The most frequent complications were transient ophthalmoplegias. Minor modifications to the catheter system should reduce the incidence of such catastrophes and complications.


Energy yield of denitrification: an estimate from growth yield in continuous cultures of Pseudomonas denitrificans under nitrate-, nitrite- and oxide-limited conditions.
The molar growth yields of Pseudomonas denitrificans, for nitrate, nitrite and nitrous oxide, were determined in chemostat culture under electron acceptor-limited conditions. Glutamate was used as the source of energy, carbon and nitrogen. The catabolic pattern was identical, irrespective of the terminal electron acceptors. The molar growth yields, corrected for maintenance energy, were 28-6 g/mol nitrate, 16-9 g/mol nitrite and 8-8 g/mol nitrous oxide. The energy yield, expressed on an electron basis, was proportional to the oxidation number of the nitrogen: nitrate (plus 5), nitrite (plus 3) and nitrous oxide (plus 1). It was concluded that oxidative phosphorylation occurs to a similar extent in each of the electron transport chains associated with the reduction of nitrate to nitrite, nitrite to nitrous oxide and nitrous oxide to nitrogen.


[Status of the knowledge of the Cynipini (Hymenoptera: Cynipidae) in Mexico: perspectives for future studies].
The knowledge status of the gall-former cynipids in the Mexican oaks is commented herein. For each of the 157 cited species from 33 oak species, the most important biological and ecological information is included. Andricus championi (Cameron), a species described only from the gall, is considered as 'incertae sedis'. Andricus tumeralis Pujade-Villar nom. nov. is proposed to denominate the species Andricus ashmeadi Dalla Torre & Kieffer for being a homonymy of Andricus ashmeadi Basset. Atrusca dugesi (Mayr) n. comb. (formery in Cynips) and Antron sagata (Kinsey) n. comb. (formery in Sphaeroteras) are proposed. The reasons of these changes are commented as well as the need for further studies on Mexican cynipids.


High expression of calreticulin indicates poor prognosis and modulates cell migration and invasion via activating Stat3 in nasopharyngeal carcinoma.
Objective: Emerging evidence suggests that calreticulin (CALR) has great impacts on the tumor formation and progression of various cancers, but the role of CALR remains controversial. We investigated the expression and clinical significance of CALR in nasopharyngeal carcinoma (NPC). Methods: Immunohistochemistry was used to detect the expression of CALR in NPC tissues, and the correlation of CALR with clinicopathological characteristics and prognosis were analyzed. The cell functions of CALR in NPC cells were also performed in vitro. Results: Compared with non-tumor nasopharyngeal epithelium (NPE) tissues, CALR expression was markedly up-regulated in NPC tissues (P < 0.001), and the high expression of CALR was positively associated with advanced clinical stage (P=0.003) and metastasis (P=0.023). Compared to the patients with low expression of CALR, patients who displayed high expression of CALR may achieve a poorer progression-free survival (PFS) and overall survival (OS) (P < 0.001). Furthermore, multivariate analysis showed that high expression of CALR was an independent predictor of poor prognosis. In addition, we found that knockdown of CALR significantly inhibited the proliferation, migration and invasion of CNE2 and HONE1 cells in vitro, and the mechanism might be associated with inactivation of Stat3 signaling pathway. Conclusion: CALR may promote NPC progression and metastasis via involving Stat3 signaling pathway, and can be regarded as an effective potential predictor for progression and prognosis of NPC.


Will an imperfect vaccine curtail the COVID-19 pandemic in the U.S.?
The novel coronavirus (COVID-19) that emerged from Wuhan city of China in late December 2019 continue to pose devastating public health and economic challenges across the world. Although the community-wide implementation of basic non-pharmaceutical intervention measures, such as social distancing, quarantine of suspected COVID-19 cases, isolation of confirmed cases, use of face masks in public, contact tracing and testing, have been quite effective in curtailing and mitigating the burden of the pandemic, it is universally believed that the use of a vaccine may be necessary to effectively curtail and eliminating COVID-19 in human populations. This study is based on the use of a mathematical model for assessing the impact of a hypothetical imperfect anti-COVID-19 vaccine on the control of COVID-19 in the United States. An analytical expression for the minimum percentage of unvaccinated susceptible individuals needed to be vaccinated in order to achieve vaccine-induced community herd immunity is derived. The epidemiological consequence of the herd immunity threshold is that the disease can be effectively controlled or eliminated if the minimum herd immunity threshold is achieved in the community. Simulations of the model, using baseline parameter values obtained from fitting the model with COVID-19 mortality data for the U.S., show that, for an anti-COVID-19 vaccine with an assumed protective efficacy of 80%, at least 82% of the susceptible US population need to be vaccinated to achieve the herd immunity threshold. The prospect of COVID-19 elimination in the US, using the hypothetical vaccine, is greatly enhanced if the vaccination program is combined with other interventions, such as face mask usage and/or social distancing. Such combination of strategies significantly reduces the level of the vaccine-induced herd immunity threshold needed to eliminate the pandemic in the US. For instance, the herd immunity threshold decreases to 72% if half of the US population regularly wears face masks in public (the threshold decreases to 46% if everyone wears a face mask).


The role of dysregulated glucose metabolism in epithelial ovarian cancer.
Epithelial ovarian cancer (EOC) is the most lethal gynecologic cancer and also one of the most poorly understood. Other health issues that are affecting women with increasing frequency are obesity and diabetes, which are associated with dysglycemia and increased blood glucose. The Warburg Effect describes the ability of fast-growing cancer cells to preferentially metabolize glucose via anaerobic glycolysis rather than oxidative phosphorylation. Recent epidemiological studies have suggested a role for hyperglycemia in the pathogenesis of a number of cancers. If hyperglycemia contributes to tumour growth and progression, then it is intuitive that antihyperglycemic drugs may also have an important antitumour role. Preliminary reports suggest that these drugs not only reduce available plasma glucose, but also have direct effects on cancer cell viability through modification of molecular energy-sensing pathways. This review investigates the effect that hyperglycemia may have on EOC and the potential of antihyperglycemic drugs as therapeutic adjuncts.


A kind of integrated serial algorithms for noise reduction and characteristics expanding in respiratory sound.
The computer‑based lung respiratory sound analysis, which can provide more information about the condition of lung station, has achieved a great development in recent years. However, the external noise in respiratory sound signal is a large restriction to the further promotion of this technique. In this paper, a kind of serial integrated de-noising algorithms which consist of a FIR band-pass filter and a modified wavelet filter and an adaptive filter, is proposed to suppress the noise in respiratory sound signals. The design of this kind of filter and its practical application are studied. The practical application in de-noise of the lung sound shows that this filter has a good de-noising effect and a good performance in outstanding the acoustic characteristics.


The evolution and rise of stereotactic body radiotherapy (SBRT) for spinal metastases.
Owing to improvements in clinical care and systemic therapy, more patients are being diagnosed with, and living longer with, spinal metastases (SM). In parallel, tremendous technological progress has been made in the field of radiation oncology. Advances in both software and hardware are able to integrate three- (and four-) dimensional body imaging with spatially accurate treatment delivery methods. This leads to improved efficacy, shortened treatment schedule, and potentially reduced treatment-related toxicity. Areas covered: In this review, we will look at the progress made by stereotactic body radiotherapy (SBRT) in the management of SM. We will review the technological factors which have enabled the widespread use of SBRT. The efficacy of SBRT, in various clinical scenarios, and associated toxicities will be reviewed. Lastly, we will discuss about patient selection and provide a five-year roadmap. Expert commentary: Spine SBRT is a safe and efficacious treatment option. Practice guidelines recommend the use of SBRT in oligometastatic patients especially those with radio-resistant cancer types, and in scenarios involving re-irradiation. SBRT offers patients dose-intensification over a short schedule which may allow less time off systemic therapy. The results of the phase III trials are eagerly awaited.


Survival differences between organ preservation surgery and definitive radiotherapy in early supraglottic squamous cell carcinoma.
Single-modality treatment, either with organ preservation surgery (OPS) or definitive radiation (RT), is the treatment of choice for patients with early supraglottic squamous cell carcinoma (SGC). However, studies comparing the effectiveness of these 2 techniques are lacking. This study compares the survival outcomes in early SGC patients treated with OPS versus RT. Secondary data analysis. Surveillance, Epidemiology and End Results database. This study included adult patients with early-stage (T1N0, T2N0) SGC undergoing single-modality treatment with either OPS (with or without neck dissection [ND]) or RT between 1988 and 2008. Survival analysis was used to compare the overall survival (OS) and disease-specific survival (DSS) between patients treated with OPS+ND, OPS alone, and RT. A total of 2631 T1/T2 N0 SGC patients were identified, of whom 167 (6%) were treated with OPS+ND, 186 (7%) with OPS only, and 2278 patients (87%) with definitive RT only. In stage I (T1N0) SGC patients, a significantly better 5-year DSS was noted for both OPS+ND (81% vs 68%, hazard ratio [HR] = 0.61, P = .03) and OPS only (82% vs 68%, HR = 0.70, P = .05) when compared with definitive RT. For stage II (T2N0) patients, only OPS+ND resulted in a significantly better 5-year DSS (86% vs 60%, HR = 0.31, P < .001) when compared with patients treated with RT. Patients with early SGC who underwent OPS+ND had better OS and DSS than patients undergoing RT alone. OPS+ND may be considered a viable and preferred treatment option in these patients.


Reactivity of serum antibodies to the keratin layer of rat esophagus in patients with rheumatoid arthritis.
Serum antibodies reactive with the keratin layer of rat esophagus (AKA) were found in 46 of 80 (57.5%) rheumatoid arthritis (RA) patients. In contrast, AKA were present in only 7 of 82 (9.5%) patients with other types of rheumatic disorders and in 2 of 47 (4.2%) healthy subjects. AKA were not specific for RA, however, because in the former group, AKA were present in 4 of 20 (20%) systemic sclerosis patients and in 3 of 12 (25%) ankylosing spondylitis patients. AKA belong predominantly to the IgG class and are complement fixing. Although found in some RA joint fluids, AKA were not selectively concentrated in the joint fluid. Absorption of RA serum with type I human collagen or with human epidermal keratin did not remove AKA activity. The frequency of AKA in RA patients both negative and positive for DR4 was equal. There was no relationship between the frequency of AKA and the occurrence of other serum autoantibodies such as antibodies to intermediate filaments, smooth muscle, and nuclear antigens. Serum antibody reactive with human stratum corneum found in patients with psoriatic arthritis was shown to be different from AKA. Rabbit antiserum to human keratin did not inhibit the reaction of AKA against the keratin layer of rat esophagus. Autoimmunity to structural proteins including collagen, vimentin intermediate filaments, smooth muscle antigens, and keratin is a characteristic feature of RA.


Fibreoptic examination of the colon: a review of 255 cases.
The results of fibreendoscopy of the colon are described in 255 consecutive examinations. Of the examinations, 26.5% resulted in a diagnosis being made solely by endoscopy. This included 17 cases of carcinoma, 15 patients with polyps, and 25 patients with inflammatory bowel disease. The most common reason for referral was undiagnosed rectal bleeding (75 cases) and endoscopy alone was successful in diagnosing the probable or definitive source of bleeding in 50% of the cases referred. It is not yet clear, however, to what extent this figure may reflect the inadequacy of conventional radiology.A definitive radiological diagnosis was refuted in 11 patients and an unnecessary laparotomy avoided in seven of these. There is no doubt that fibreoptic colonoscopy increases diagnostic accuracy in large bowel disease and is especially helpful in cases where radiology is either negative or equivocal.


Monitoring water content in deforming intervertebral disc tissue by finite element analysis of MRI data.
Mechanical loading, occurring during normal daily life, causes fluid to be expelled from intervertebral discs. Excessive fluid loss during heavy loading might make the disc more vulnerable to damage. In this study, fluid loss was investigated in vitro through monitoring the loss of MRI signal intensity in four bovine coccygeal intervertebral discs, compressed at 2000 N during 1.5 hr. The MRI signals were analyzed with the aid of finite element models to account for the deformation of the tissue. A gradual signal loss over time was found during loading, the most pronounced loss occurring in the central disc region. Initial patterns of signal distribution were quite variable between specimens but repeatable within specimens.


Distribution of troponin components in the thin filament studied by immunoelectron microscopy.
1. The localization of specific antiboidies against troponin components, i.e., troponin T(TN-T), troponin I(TN-I, and troponin C(TN-C), was studied by the use of an electrom microscope. 2. Every antibody was distributed along the thin filament with a period of 38 nm. 3. Staining with anti-TN-I or anti-TN-C formed narrow striations. The location of the first striation was 26 nm from the free end of the thin filament. 4. The width of individual striations formed by anti-TN-T was 14--20nm The H-band-side end of each striation coincided with the location of anti-TN-I or anti-TN-C.


A comparison of birth weight and weight/length ratio for gestation as correlates of perinatal morbidity.
The purpose was to evaluate a low weight to length ratio as a correlate of perinatal morbidity and mortality. Data from the Collaborative Perinatal Project for infants of 34 weeks' gestation or more were evaluated. Associations between the weight to length ratio of < 10% (low weight to length) and birth weight of < 10% (small for gestational age) by gestational age and gender, perinatal depression, dysmaturity, cerebral palsy, and neonatal mortality were evaluated. A low weight to length ratio and small for gestational age status were associated with most markers of perinatal morbidity and mortality in term and preterm infants. In infants not small for gestational age, a low weight to length ratio was associated with increased morbidity and mortality (relative risk of 1.9 to 4.2) in term infants, and with perinatal depression (relative risk of 2.9) in preterm infants. Logistic regression found low weight to length ratio was a better independent correlate than small for gestational age status for all markers assessed and found low weight to length ratio was significantly associated with all morbidity and mortality markers in infants not small for gestational age. Low weight to length ratio, a marker for asymmetric growth restriction, is correlated with perinatal morbidity, even in infants not small for gestational age.


Chronic prostatic infection and inflammation by Propionibacterium acnes in a rat prostate infection model.
Chronic inflammation in the prostate, seen as infiltration of inflammatory cells into the prostate gland in histological samples, affects approximately half the male population without indication of prostate disease, and is almost ubiquitous in patients diagnosed with benign prostate hyperplasia and cancer. Several studies have demonstrated the gram-positive bacterium Propionibacterium acnes to be frequently present in prostate tissue from men suffering from prostate disease. P. acnes has been shown to be associated with histological inflammation in human prostatectomy specimens, and also to induce strong inflammatory response in prostate-derived tissue culture models. The present paper describes a rat model for assessment of the pathogenic potential of P. acnes in prostate. Prostate glands of Sprague Dawley rats (n = 98) were exposed via an abdominal incision and live P. acnes or, in control rats, saline were injected into the ventral and dorso-lateral lobes. Rats were sacrificed 5 days, 3 weeks, 3 months and 6 months post infection, and prostate tissue was analyzed for bacterial content and histological inflammation. Rat sera were assessed for levels of CRP and anti-P. acnes IgG. Live P. acnes could be recovered from the dorso-lateral lobes up to 3 months post infection, while the ventral lobes were cleared from bacteria at that time. In samples up to 3 months post infection, the dorso-lateral lobes exhibited intense focal inflammation. CRP and IgG levels were elevated throughout the span of the experiment, and reached maximum levels 3 weeks and 3 months post infection, respectively. We show that P. acnes have the potential to cause chronic infection in previously healthy prostate, and that the infection has potential to cause chronic histological inflammation in the infected tissue. The high prevalence of P. acnes in human prostate tissue calls for resolution of pathogenic details. The present rat model suggests that complications such as chronic inflammation may be induced by P. acnes infection.


Quantitative analysis of gene expression in preimplantation mouse embryos using green fluorescent protein reporter.
We have developed a method to monitor noninvasively, quantitatively, and in real-time transcription in living preimplantation mouse embryos by measuring expression of a short half-life form of enhanced green fluorescent protein (EGFP) following microinjection of a plasmid-borne EGFP reporter gene. A standard curve was established by injecting known amounts of recombinant green fluorescent protein, and transcriptional activity was then determined by interpolating the amount of fluorescence in the DNA-injected embryos. This approach permitted multiple measurements in single embryos with no significant detrimental effect on embryonic development as long as light exposure was brief (<30 sec) and no more than two measurements were made each day. This method should facilitate analysis of the regulation of gene expression in preimplantation embryos; in particular, during the maternal-to-zygotic transition, and in other species in which limited numbers of embryos are available.


Nuclear DNA analyses in genetic studies of populations: practice, problems and prospects.
Population-genetic studies have been remarkably productive and successful in the last decade following the invention of PCR technology and the introduction of mitochondrial and microsatellite DNA markers. While mitochondrial DNA has proven powerful for genealogical and evolutionary studies of animal populations, and microsatellite sequences are the most revealing DNA markers available so far for inferring population structure and dynamics, they both have important and unavoidable limitations. To obtain a fuller picture of the history and evolutionary potential of populations, genealogical data from nuclear loci are essential, and the inclusion of other nuclear markers, i.e. single copy nuclear polymorphic (scnp) sequences, is clearly needed. Four major uncertainties for nuclear DNA analyses of populations have been facing us, i.e. the availability of scnp markers for carrying out such analysis, technical laboratory hurdles for resolving haplotypes, difficulty in data analysis because of recombination, low divergence levels and intraspecific multifurcation evolution, and the utility of scnp markers for addressing population-genetic questions. In this review, we discuss the availability of highly polymorphic single copy DNA in the nuclear genome, describe patterns and rate of evolution of nuclear sequences, summarize past empirical and theoretical efforts to recover and analyse data from scnp markers, and examine the difficulties, challenges and opportunities faced in such studies. We show that although challenges still exist, the above-mentioned obstacles are now being removed. Recent advances in technology and increases in statistical power provide the prospect of nuclear DNA analyses becoming routine practice, allowing allele-discriminating characterization of scnp loci and microsatellite loci. This certainly will increase our ability to address more complex questions, and thereby the sophistication of genetic analyses of populations.


Enabling LIFDI-MS measurements of highly air sensitive organometallic compounds: a combined MS/glovebox technique.
A new setup combining a ThermoFisher Exactive Plus Orbitrap Mass Spectrometer with a liquid injection field desorption ionization (LIFDI) source directly connected to an inert atmosphere glovebox is presented. The described setup allows for the analysis of very air- and moisture sensitive samples. Furthermore, the soft nature of LIFDI ionization gives access to the molecular ions of fragile molecules. This new setup is therefore especially useful for sensitive organometallic complexes. The functionality of the new setup is tested against [(Cp)2TiCl]˙, which is known for its notorious sensitivity to air and moisture. Its drastic colour change from green to orange upon exposure to air further supports the easy detection of traces of oxygen during the experiment. In addition, we applied this setup to the mass spectrometric analysis of the qualitative composition of a Cu/Al cluster mixture, which is not accessible by other analytical methods.


Chemical process synthesis of beta-amino acids and esters.
The importance of beta-amino acids and esters for the synthesis of potential therapeutic agents and biologically active compounds is well known and the subject of this special issue. This review outlines some of the recent approaches reported for the synthesis of both racemic and enantiomeric beta-amino acids and esters with emphasis on those used for large scale production. This compilation is written from a chemical process perspective focusing on the practical application of these processes for large scale synthesis. A survey of procedures described in patent publications as well as current advances from chemical process research groups and results from our laboratory are included with emphasis on asymmetric Michael-type additions, addition of metal enolates to imines/sulfinimines, classical and enzymatic resolutions, and reduction of enantiomeric enamines.


Transforming growth factor alpha (TGF(alpha)) is increased during hyperoxia and fibrosis.
Transforming growth factor alpha (TGF(alpha)) stimulates type II alveolar epithelial cell proliferation and also is associated with fibrosis. We studied the changes in bronchoalveolar lavage (BAL) TGF(alpha) protein in a neonatal rabbit hyperoxia-fibrosis model (100% O(2) for 8 to 9 days, followed by 60% O(2) to 36 days of age). Hyperoxia increased TGF(alpha) protein and delayed the appearance of mature lower molecular weight (MW) TGF(alpha) isoforms at postnatal days 6 and 8 during the acute injury period. At 3 and 5 weeks, after chronic hyperoxia exposure, there was an increase in lower MW TGF(alpha) peptides during the fibrotic period. Keratinocyte growth factor (KGF) is also a type II cell mitogen. In vitro studies of keratinocytes suggest that KGF-induced proliferation is mediated through TGF(alpha). Intratracheal KGF instillation into adult wild-type and TGF(alpha)-null mice demonstrated that the KGF induced equivalent robust levels of proliferation in both TGF(alpha) deficient and wild-genotype mice. In conclusion, there are both quantitative and qualitative changes in TGF(alpha) protein in a hyperoxia-induced fibrosis neonatal rabbit model during periods of type II cell proliferation and fibrosis.


Dual actions of the Galpha(q) agonist Pasteurella multocida toxin to promote cardiomyocyte hypertrophy and enhance apoptosis susceptibility.
Previous attempts to delineate the consequences of Galpha (q) activation in cardiomyocytes relied largely on molecular strategies in cultures or transgenic mice. Modest levels of wild-type Galpha(q) overexpression induce stable cardiac hypertrophy, whereas intense Galpha(q) stimulation induces cardiomyocyte apoptosis. The precise mechanism(s) whereby traditional targets of Galpha (q) subunits that induce hypertrophy also trigger cardiomyocyte apoptosis is not obvious and is explored with recombinant Pasteurella multocida toxin (rPMT, a Galpha(q) agonist). Cells cultured with rPMT display cardiomyocyte enlargement, sarcomeric organization, and increased atrial natriuretic factor expression in association with activation of phospholipase C, novel protein kinase C (PKC) isoforms, extracellular signal-regulated protein kinase (ERK), and (to a lesser extent) JNK/p38-MAPK. rPMT stimulates the ERK cascade via epidermal growth factor (EGF) receptor transactivation in cardiac fibroblasts, but EGF receptor transactivation plays no role in ERK activation in cardiomyocytes. Surprisingly, rPMT (or novel PKC isoform activation by PMA) decreases basal Akt phosphorylation; rPMT prevents Akt phosphorylation by EGF or IGF-1 and functionally augments cardiomyocyte apoptosis in response to H2O2. These results identify a Galpha(q)-PKC pathway that represses basal Akt phosphorylation and impairs Akt stimulation by survival factors. Because inhibition of Akt enhances cardiomyocyte susceptibility to apoptosis, this pathway is predicted to contribute to the transition from hypertrophy to cardiac decompensation and could be targeted for therapy in heart failure.


Structural basis for the ABO blood-group dependence of Plasmodium falciparum rosetting.
The ABO blood group influences susceptibility to severe Plasmodium falciparum malaria. Recent evidence indicates that the protective effect of group O operates by virtue of reduced rosetting of infected red blood cells (iRBCs) with uninfected RBCs. Rosetting is mediated by a subgroup of PfEMP1 adhesins, with RBC binding being assigned to the N-terminal DBL1α₁ domain. Here, we identify the ABO blood group as the main receptor for VarO rosetting, with a marked preference for group A over group B, which in turn is preferred to group O RBCs. We show that recombinant NTS-DBL1α₁ and NTS-DBL1α₁-CIDR1γ reproduce the VarO-iRBC blood group preference and document direct binding to blood group trisaccharides by surface plasmon resonance. More detailed RBC subgroup analysis showed preferred binding to group A₁, weaker binding to groups A₂ and B, and least binding to groups A(x) and O. The 2.8 Å resolution crystal structure of the PfEMP1-VarO Head region, NTS-DBL1α₁-CIDR1γ, reveals extensive contacts between the DBL1α₁ and CIDR1γ and shows that the NTS-DBL1α₁ hinge region is essential for RBC binding. Computer docking of the blood group trisaccharides and subsequent site-directed mutagenesis localized the RBC-binding site to the face opposite to the heparin-binding site of NTS-DBLα₁. RBC binding involves residues that are conserved between rosette-forming PfEMP1 adhesins, opening novel opportunities for intervention against severe malaria. By deciphering the structural basis of blood group preferences in rosetting, we provide a link between ABO blood grouppolymorphisms and rosette-forming adhesins, consistent with the selective role of falciparum malaria on human genetic makeup.


Mapping the Proteoform Landscape of Five Human Tissues.
A functional understanding of the human body requires structure-function studies of proteins at scale. The chemical structure of proteins is controlled at the transcriptional, translational, and post-translational levels, creating a variety of products with modulated functions within the cell. The term "proteoform" encapsulates this complexity at the level of chemical composition. Comprehensive mapping of the proteoform landscape in human tissues necessitates analytical techniques with increased sensitivity and depth of coverage. Here, we took a top-down proteomics approach, combining data generated using capillary zone electrophoresis (CZE) and nanoflow reversed-phase liquid chromatography (RPLC) hyphenated to mass spectrometry to identify and characterize proteoforms from the human lungs, heart, spleen, small intestine, and kidneys. CZE and RPLC provided complementary post-translational modification and proteoform selectivity, thereby enhancing the overall proteome coverage when used in combination. Of the 11,466 proteoforms identified in this study, 7373 (64%) were not reported previously. Large differences in the protein and proteoform level were readily quantified, with initial inferences about proteoform biology operative in the analyzed organs. Differential proteoform regulation of defensins, glutathione transferases, and sarcomeric proteins across tissues generate hypotheses about how they function and are regulated in human health and disease.


Negative Effects of Annealed Seed Layer on the Performance of ZnO-Nanorods Based Nitric Oxide Gas Sensor.
Nitric oxide (NO) is a toxic gas, which is dangerous for human health and causes many respiratory infections, poisoning, and lung damage. In this work, we have successfully grown ZnO nanorod film on annealed ZnO seed layer in different ambient temperatures, and the morphology of the nanorods sensing layer that affects the gas sensing response to nitric oxide (NO) gas were investigated. To acknowledge the effect of annealing treatment, the devices were fabricated with annealed seed layers in air and argon ambient at 300 °C and 500 °C for 1 h. To simulate a vertical device structure, a silver nanowire electrode covered in ZnO nanorod film was placed onto the hydrothermal grown ZnO nanorod film. We found that annealing treatment changes the seed layer's grain size and defect concentration and is responsible for this phenomenon. The I-V and gas sensing characteristics were dependent on the oxygen defects concentration and porosity of nanorods to react with the target gas. The resulting as-deposited ZnO seed layer shows better sensing response than that annealed in an air and argon environment due to the nanorod morphology and variation in oxygen defect concentration. At room temperature, the devices show good sensing response to NO concentration of 10 ppb and up to 100 ppb. Shortly, these results can be beneficial in the NO breath detection for patients with chronic inflammatory airway disease, such as asthma.


Sedimentology and chemostratigraphy of the terminal Ediacaran Dengying Formation at the Gaojiashan section, South China.
The late Ediacaran Dengying Formation (ca. 551.1-538.8 Ma) in South China is one of two successions where Ediacara-type macrofossils are preserved in carbonate facies along with skeletal fossils and bilaterian animal traces. Given the remarkable thickness of carbonate-bearing strata deposited in less than 12.3 million years, the Dengying Formation holds the potential for a relatively continuous chemostratigraphic profile for the terminal Ediacaran stage. In this study, a detailed sedimentological and chemostratigraphic (δ13Ccarb, δ18Ocarb, δ13Corg, δ34Spyrite, and 87Sr/86Sr) investigation was conducted on the Dengying Formation at the Gaojiashan section, Ningqiang County of the southern Shaanxi Province, South China. Sedimentological results reveal an overall shallow marine depositional environment. Carbonate breccia, void-filling botryoidal precipitates, and aragonite crystal fans are common in the Algal Dolomite Member of the Dengying Formation, suggesting that peritidal facies were repeatedly karstified. The timing of karstification was likely early, probably soon after the deposition of the dolomite sediments. The presence of authigenic aragonite cements suggests high alkalinity in the terminal Ediacaran ocean. Geochemical analysis of micro-drilled samples shows that distinct compositions are registered in different carbonate phases, which should be considered when constructing chemostratigraphic profiles representative of true temporal variations in seawater chemistry. Integrated chemostratigraphic data suggest enhanced burial of organic carbon and pyrite, and the occurrence of extensive marine anoxia (at least in the Gaojiashan Member). Rapid basinal subsidence and carbonate accumulation during a time of elevated seawater alkalinity and increased rates of pyrite burial may have facilitated the evolutionary innovation of early biomineralizing metazoans.


Tunable Dynamics in a Multistrain Transcriptional Pulse Generator.
One challenge in synthetic biology is the tuning of regulatory components within gene circuits to elicit a specific behavior. This challenge becomes more difficult in synthetic microbial consortia since each strain's circuit must function at the intracellular level and their combination must operate at the population level. Here we demonstrate that circuit dynamics can be tuned in synthetic consortia through the manipulation of strain fractions within the community. To do this, we construct a microbial consortium comprised of three strains of engineered Escherichia coli that, when cocultured, use homoserine lactone-mediated intercellular signaling to create a multistrain incoherent type-1 feedforward loop (I1-FFL). Like naturally occurring I1-FFL motifs in gene networks, this engineered microbial consortium acts as a pulse generator of gene expression. We demonstrate that the amplitude of the pulse can be easily tuned by adjusting the relative population fractions of the strains. We also develop a mathematical model for the temporal dynamics of the microbial consortium. This model allows us to identify population fractions that produced desired pulse characteristics, predictions that were confirmed for all but extreme fractions. Our work demonstrates that intercellular gene circuits can be effectively tuned simply by adjusting the starting fractions of each strain in the consortium.


Malignancy Rates of Thyroid Cytology: Cyst Fluid Benign or Non-Diagnostic?
BACKGROUND We sought to investigate subgroup distribution using Bethesda classification and risks for malignancy. We also compared the malignancy risk of cases that were denoted as non-diagnostic due to cystic contents, with cases that were denoted as non-diagnostic due to presence of other features. MATERIAL AND METHODS The study included pathology test results of 1,440 thyroid nodule samples diagnosed using Bethesda classification. Results of 305 thyroidectomy excision specimens from these patients were also compared with cytology results to determine the frequency of malignancy. The non-diagnostic group was divided into two categories: those with cystic contents, and others. Malignancy rates were separately calculated for the two groups, and compared with the other classification groups. RESULTS Distribution of malignancy rates by Bethesda classification were as follows: non-diagnostic 12.5% (6/48), benign 1.5% (3/198), atypia of undetermined significance/follicular lesion of undetermined significance (AFLUS) 9% (1/11), suspicious for follicular neoplasm (SFN) 37.5% (3/8), suspicious malignancy 70% (8/26), malignancy 100% (14/14). CONCLUSIONS Despite the limited number of cases, our study concluded that cystic content was closer to the benign category than the non-diagnostic category if the assessment was based on malignancy rates. In this group, similar to aspirations containing plenty of lymphocytes that indicates colloid or lymphocytic thyroiditis, it is still controversial whether criterion for adequacy of follicular epithelial cells should be sought, or if they should be regarded as benign in order to prevent unnecessarily performance of repeat aspirations.


MicroRNA 29 targets nuclear factor-κB-repressing factor and Claudin 1 to increase intestinal permeability.
Some patients with irritable bowel syndrome with diarrhea (IBS-D) have intestinal hyperpermeability, which contributes to their diarrhea and abdominal pain. MicroRNA 29 (MIR29) regulates intestinal permeability in patients with IBS-D. We investigated and searched for targets of MIR29 and investigated the effects of disrupting Mir29 in mice. We investigated expression MIR29A and B in intestinal biopsies collected during endoscopy from patients with IBS (n = 183) and without IBS (controls) (n = 36). Levels were correlated with disease phenotype. We also generated and studied Mir29(-/-) mice, in which expression of Mir29a and b, but not c, is lost. Colitis was induced by administration of 2,4,6-trinitrobenzenesulfonic acid; intestinal tissues were collected and permeability was assessed. Microarray analysis was performed using tissues from Mir29(-/-) mice. Changes in levels of target genes were measured in human colonic epithelial cells and small intestinal epithelial cells after knockdown of MIR29 with anti-MIRs. Intestinal tissues from patients with IBS-D (but not IBS with constipation or controls) had increased levels of MIR29A and B, but reduced levels of Claudin-1 (CLDN1) and nuclear factor-κB-repressing factor (NKRF). Induction of colitis and water avoidance stress increased levels of Mir29a and Mir29b and intestinal permeability in wild-type mice; these increased intestinal permeability in colons of far fewer Mir29(-/-) mice. In microarray and knockdown experiments, MIR29A and B were found to reduce levels of NKRF and CLDN1 messenger RNA, and alter levels of other messenger RNAs that regulate intestinal permeability. Based on experiments in knockout mice and analyses of intestinal tissue samples from patients with IBS-D, MIR29 targets and reduces expression of CLDN1 and NKRF to increase intestinal permeability. Strategies to block MIR29 might be developed to restore intestinal permeability in patients with IBS-D.


Essential thrombocytosis with the Philadelphia chromosome (Ph').
Essential thrombocytosis is a myeloproliferative disease not known to have consistent cytogenetic abnormalities. A 46-year-old black woman with essential thrombocytosis and a Philadelphia chromosome is reported. Iron deficiency and tuberculosis were present but when effectively treated did not result in resolution of thrombocytosis. Megakaryocytic hyperplasia of bone marrow, abnormal platelet function studies and a compatible clinical state suggested the diagnosis of essential thrombocytosis. The diagnostic criteria for other myeloproliferative diseases were not met. The Philadelphia chromosome was consistently obtained from bone marrow preparations. We conclude that the Philadelphia chromosome may be found in essential thrombocytosis as well as other, previously reported, myeloproliferative diseases.


CD4+ CD8+ thymocytes are preferentially induced to die following CD45 cross-linking, through a novel apoptotic pathway.
Ligation of the protein tyrosine phosphatase CD45 on both mature and immature T cells modulates the amplitude of TCR-mediated signals. In this work, we have evaluated the consequences of CD45 ligation on immature T cells, in the absence of TCR engagement. Cross-linking of CD45 on thymocytes by mAbs led to the induction of cellular death, characterized by a reduction in mitochondrial membrane potential (delta psi(m)), production of reactive oxygen species, loss in membrane asymmetry, exposure of phosphatidylserine residues, and incorporation of vital dyes. In sharp contrast to most stimuli causing thymocyte death, CD45 cross-linking did not lead to DNA degradation. Cell death was not blocked by Bcl-2 overexpression or treatment with caspase inhibitor. However, death was inhibited by the addition of scavengers of reactive oxygen species. We also established that susceptibility to CD45-mediated death is acquired during the transition of early CD4- CD8- TCR- T cell precursors into CD4+ CD8+ TCR- thymocytes and is increased with further acquisition of surface TCR on these cells. Moreover, mature thymocytes were much less sensitive to CD45 cross-linking than CD4+ CD8+ cells. We propose that during T cell development, CD45 ligation could induce the death of those immature thymocytes that do not fulfill the requirements for positive selection.