Three approaches to regression analysis of receiver operating characteristic curves for continuous test results.
The accuracy of a medical diagnostic test is typically summarized by the sensitivity and specificity when the test result is dichotomous. Receiver operating characteristic (ROC) curves are measures of test accuracy that are used when test results are continuous and are considered the analogs of sensitivity and specificity for continuous tests. ROC regression analysis allows one to evaluate effects of factors that may influence test accuracy. Such factors might include characteristics of study subjects or operating conditions for the test. Unfortunately, regression analysis methods for ROC curves are not well developed and methods that do exist have received little use to date. In this paper, we propose and compare three very different regression analysis methods. Two are modifications of methods previously proposed for radiology settings. The third is a special case of a general method recently proposed by us. The three approaches are compared with regard to settings in which they can be applied and distributional assumptions they require. In the setting where test results are normally distributed, we elucidate the correspondence between regression parameters in the different models. The methods are applied to simulated data and to data from a study of a new diagnostic test for hearing impairment. It is hoped that the presentation in this paper will both encourage the use of regression analysis for evaluating diagnostic tests and help guide the choice of the most appropriate regression analysis approach in applications.


Cardiovascular responses to stimulation of pulmonary C fibres in the cat: their modulation by changes in respiration.
1. In cats anaesthetized with a mixture of chloralose and urethane, stimulation of pulmonary C fibres by right atrial injections of phenylbiguanide caused, after a latency of about 3 s, a reduction in pulmonary ventilation or apnoea, bradycardia and systemic hypotension, confirming previous work. 2. A decrease in femoral artery perfusion pressure also occurred under conditions in which the hindlimb was vascularly isolated, the blood flow was maintained constant and the inferior vena caval pressure did not change. This indicates a reduction in vascular resistance due to vasodilatation. The response was unaffected by atropine and propranolol, but was reduced or abolished by guanethidine, hexamethonium and denervation of the limb, indicating that it is due to a reduction in activity in sympathetic vasoconstrictor fibres. 3. Similar cardiovascular responses were observed when the arterial blood pressure was maintained constant, and also in artificially ventilated animals. 4. Evidence is presented that the receptors responsible for the respiratory and cardiovascular responses to right atrial injections of phenylbiguanide lie in the pulmonary vascular bed. 5. When the pulmonary C fibres were excited during a period of apnoea which was induced reflexly by electrical stimulation of the central end of a superior laryngeal nerve, there were no consistent differences in the size of the cardiac or vascular responses compared to the control responses in the absence of the laryngeal input. This result occurred irrespective of the size of the control ventilatory response to phenylbiguanide. 6. By contrast in the same experiments, the cardio-inhibitory and vasoconstrictor responses to excitation of the carotid body chemoreceptors were invariably potentiated by electrical stimulation of a superior laryngeal nerve, as found previously. 7. The possible central mechanisms responsible for the differential modulation of pulmonary C fibre and carotid chemoreceptor reflexes by respiration are discussed.


Monitoring patients with continuous-flow ventricular assist devices outside of the intensive care unit: novel challenges to bedside nursing.
Ventricular assist devices provide therapeutic options for patients with severe heart failure who have exhausted available medical therapies. With restoration of organ perfusion with ventricular assist devices, the heart failure resolves and quality of life and functional status improve. The current generation of continuous-flow devices present novel challenges to the clinical assessment of patients by substantially reducing or nearly eliminating any palpable pulse. Patients therefore generally have inadequate arterial pulsatility for most noninvasive monitoring devices such as pulse oximeters or automated blood pressure cuffs to work accurately. This article describes the function of continuous-flow devices and how this function affects common monitoring options, as well as how to clinically assess recipients of continuous-flow devices to promptly identify those whose condition may be deteriorating or who may be receiving inadequate perfusion.


Novel technique for quantifying adhesion of Metarhizium anisopliae conidia to the tick cuticle.
The present study describes an accurate quantitative method for quantifying the adherence of conidia to the arthropod cuticle and the dynamics of conidial germination on the host. The method was developed using conidia of Metarhizium anisopliae var. anisopliae (Metschn.) Sorokin (Hypocreales: Clavicipitaceae) and engorged Rhipicephalus annulatus (Say) (Arachnida: Ixodidae) females and was also verified for M. anisopliae var. acridum Driver et Milner (Hypocreales: Clavicipitaceae) and Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) larvae. This novel method is based on using an organic solvent (dichloromethane [DCM]) to remove the adhered conidia from the tick cuticle, suspending the conidia in a detergent solution, and then counting them using a hemocytometer. To confirm the efficacy of the method, scanning electron microscopy (SEM) was used to observe the conidial adherence to and removal from the tick cuticle. As the concentration of conidia in the suspension increased, there were correlating increases in both the number of conidia adhering to engorged female R. annulatus and tick mortality. However, no correlation was observed between a tick's susceptibility to fungal infection and the amount of adhered conidia. These findings support the commonly accepted understanding of the nature of the adhesion process. The mechanism enabling the removal of the adhered conidia from the host cuticle is discussed.


[Results of treating medullary thyroid carcinoma: the differences between sporadic and inherited forms].
Medullary thyroid carcinoma (MTC) can be divided into two subgroups: sporadic or inherited. Hereditary form of MTC is often believed to be form with better prognosis than sporadic one. In this study the differences in MTC prognosis in Polish population of patients was analyzed. The group of 169 patients with MTC was examined. Hereditary cancer was stated in 48 (28%) patients. The median age of disease onset was 41 years (from 7 to 71 years). Genetic examination of RET protooncogene was performed in all patients. The calcitonin and CEA serum level analysis and radiological and radioisotopic examinations were used for monitoring of the disease course. Nineteen cases of MEN 2A syndrome, 11 cases of MEN 2B one and 18 cases of non classified familial MTC were recognized among patients with inherited MTC. Significantly lower age of disease onset in inherited MTC than in sporadic one was observed (27 years vs. 43.7 years, p < 0.001). Local or nodal recurrence was observed in 22 (13%) patients, distant metastases were stated in 21 (12%) patients. Basal or stimulated serum calcitonin level was increased in 85 (50%) patients. No significant differences between sporadic and inherited disease were observed. Eight patients died during observation, including 3 patients with sporadic MTC and 5 patients with inherited MTC. The updated 10-year survival rate was 97% in patients with sporadic MTC; in hereditary MTC it was about 20% worse. The complications related to the presence of adrenal tumors were the main reason for death in MEN2 and no significant differences in the course of MTC itself were observed.


Identification of karyopherins involved in the nuclear import of RNA exosome subunit Rrp6 in Saccharomyces cerevisiae.
The exosome is a conserved multiprotein complex essential for RNA processing and degradation. The nuclear exosome is a key factor for pre-rRNA processing through the activity of its catalytic subunits, Rrp6 and Rrp44. In Saccharomyces cerevisiae, Rrp6 is exclusively nuclear and has been shown to interact with exosome cofactors. With the aim of analyzing proteins associated with the nuclear exosome, in this work, we purified the complex with Rrp6-TAP, identified the co-purified proteins by mass spectrometry, and found karyopherins to be one of the major groups of proteins enriched in the samples. By investigating the biological importance of these protein interactions, we identified Srp1, Kap95, and Sxm1 as the most important karyopherins for Rrp6 nuclear import and the nuclear localization signals recognized by them. Based on the results shown here, we propose a model of multiple pathways for the transport of Rrp6 to the nucleus.


London University Search Instrument: a combinatorial robot for high-throughput methods in ceramic science.
This paper describes the design, construction, and operation of the London University Search Instrument (LUSI) which was recently commissioned to create and test combinatorial libraries of ceramic compositions. The instrument uses commercially available powders, milled as necessary to create thick-film libraries by ink-jet printing. Multicomponent mixtures are prepared by well plate reformatting of ceramic inks. The library tiles are robotically loaded into a flatbed furnace and, when fired, transferred to a 2-axis high-resolution measurement table fitted with a hot plate where measurements of, for example, optical or electrical properties can be made. Data are transferred to a dedicated high-performance computer. The possibilities for remote interrogation and search steering are discussed.


THE EVOLUTIONARY HISTORY OF PARTHENOGENETIC CNEMIDOPHORUS LEMNISCATUS (SAURIA, TEIIDAE). I. EVIDENCE FOR A HYBRID ORIGIN.
Chromosomes and allozymes were studied from chromosomally distinct unisexual (races B and C) and bisexual (races D and E) populations of the teiid lizard Cnemidophorus lemniscatus, and from selected outgroup taxa (C. murinus, C. nigricolor, Ameiva ameiva, and A. auberi). Karyotyping confirmed the racial identity of individuals and showed that the chromosomal composition of populations at specific localities has remained the same for 20 years. All individuals of both unisexual populations were heterozygous for a pericentric inversion that distinguishes D and E bisexuals. Also, the unisexuals were all heterozygous for 8 of 11 protein loci for which D and E were fixed or nearly fixed for different alleles. Most of these alleles represent derived states relative to the other Cnemidophorus and Ameiva analyzed, and the fixed heterozygote condition at these nine markers provides unequivocal support for the hypothesis that the unisexual C. lemniscatus arose by hybridization between ancestors genetically similar to extant D and E populations. At the remaining three loci for which D and E show fixed differences, the unisexuals were homozygous rather than heterozygous. This suggests that either (1) allozymes have been lost by mutations to null, silent, or convergent mobility states, (2) ancestral genotypes were similar to but not identical with the extant D and E races, and/or (3) limited recombination may occur between unisexual genomes. Allozyme-based genetic distances between D and E were large, suggesting that bisexual races D and E are genetically isolated; each race should be accorded full species status. This conclusion is supported by the absence of any clear biochemical evidence for their monophyly with respect to the other Cnemidophorus examined. Cladistic analyses of 17 phylogenetically informative loci revealed two equally parsimonious shortest trees, one supporting monophyly and the other paraphyly of the C. lemniscatus complex. Further testing of the monophyly of C. lemniscatus requires additional data. With the present study, the evidence that all parthenogenetic Cnemidophorus are of hybrid origin is complete.


Induction of robust immune responses against human immunodeficiency virus is supported by the inherent tropism of adeno-associated virus type 5 for dendritic cells.
The ability of adeno-associated virus serotype 1 to 8 (AAV1 to AAV8) vectors expressing the human immunodeficiency virus type 1 (HIV-1) Env gp160 (AAV-HIV) to induce an immune response was evaluated in BALB/c mice. The AAV5 vector showed a higher tropism for both mouse and human dendritic cells (DCs) than did the AAV2 vector, whereas other AAV serotype vectors transduced DCs only poorly. AAV1, AAV5, AAV7, and AAV8 were more highly expressed in muscle cells than AAV2. An immunogenicity study of AAV serotypes indicates that AAV1, AAV5, AAV7, and AAV8 vectors expressing the Env gp160 gene induced higher HIV-specific humoral and cell-mediated immune responses than the AAV2 vector did, with the AAV5 vector producing the best responses. Furthermore, mice injected with DCs that had been transduced ex vivo with an AAV5 vector expressing the gp160 gene elicited higher HIV-specific cell-mediated immune responses than did DCs transduced with AAV1 and AAV2 vectors. We also found that AAV vectors produced by HEK293 cells and insect cells elicit similar levels of antigen-specific immune responses. These results demonstrate that the immunogenicity of AAV vectors depends on their tropism for both antigen-presenting cells (such as DCs) and non-antigen-presenting cells (such as muscular cells) and that AAV5 is a better vector than other AAV serotypes. These results may aid in the development of AAV-based vaccine and gene therapy.


Retinoid feeding, hormone inhibition, and/or immune stimulation and the progression of N-methyl-N-nitrosourea-induced rat mammary carcinoma: suppression by retinoids of peptide hormone-induced tumor cell proliferation in vivo and in vitro.
Female Sprague-Dawley rats were treated at 50 and 57 days of age with 2.5 mg per 100 g body weight of N-methyl-N-nitrosourea (MNU). At 60 days of age, the animals were divided into eight groups (40 rats/group) and treated as follows: (a) controls; (b) immune stimulation (IS); (c) hormone inhibition (HI); (d) HI + IS; (e) retinyl acetate feeding (RA); (f) RA + IS; (g) RA + HI; and (h) RA + HI + IS. IS treatment was accomplished by three i.p. injections (at 1, 3, and 5 weeks after carcinogen treatment) of a mixture of cell particulate preparations from pooled MNU-induced rat mammary carcinomas and Freund's (complete) adjuvant. HI treatment consisted of daily s.c. injections of tamoxifen (12.5 to 25.0 micrograms/100 g body weight) and CB-154 (200 to 400 micrograms/100 g body weight), and RA treatment consisted of daily feeding of retinyl acetate (1.0 mM). Both RA alone and HI alone significantly (p less than 0.01 to 0.001) reduced mammary carcinoma incidence; HI treatment was significantly (p less than 0.01) more effective than RA treatment. The combination of RA + HI was significantly (p less than 0.01) superior to either treatment alone; RA + HI treatment virtually completely blocked the development of mammary carcinomas at termination of study (a total of only 2 mammary carcinomas was observed in the RA + HI groups of rats at 1 year after carcinogen treatment). IS treatments did not significantly influence mammary carcinoma incidence, either alone or in combination with RA and/or HI. Female Sprague-Dawley rats given MNU and subsequently treated daily for 4 weeks with the prolactin secretion-stimulating drug haloperidol (0.05 mg/100 g body weight) responded with a significant (p less than 0.001) increase in mammary carcinoma development when compared with control rats. RA treatment of haloperidol-treated rats significantly (p less than 0.001) blocked the stimulatory effect of haloperidol on mammary carcinoma development. The addition of insulin (5.0 micrograms/ml) to the culture media of 2-day organ cultures of MNU-induced rat mammary carcinomas resulted in a significant (p less than 0.05) stimulation of [3H] thymidine incorporation into DNA of the cultured cells.(ABSTRACT TRUNCATED AT 400 WORDS)


N-terminal amino-sequencing of camel (Camelus dromedarius) luteinizing-hormone alpha and beta-subunits.
The subunits of luteinizing hormone from the Camelus dromedarius (CamLH) have been separated by reverse phase HPLC after reduction and alkylation of their disulfide bridges. The N-terminal amino-acid sequencing of the alpha and beta subunits has been performed up to the 53rd and 67th residue respectively (i.e. more than half of each polypeptide chain). These sequences have been compared to those of LH from other mammalian species in order to estimate the phylogenetic divergence of LH in this species and in order to point out characteristic features of its primary structure that can be related to its physico-chemical properties.


Effect of flaxseed oil fortified with vitamin E and phytosterols on antioxidant defense capacities and lipids profile in rats.
The main proposal of this study was to evaluate in vivo whether flaxseed oils fortified with micronutrients would have beneficial effects on lipid profile and antioxidant status in high-fat fed rats. Male Wistar rats were fed synthetic diets containing 10% of flaxseed oil (FO), phytosterols (PS) fortified FO (PS-FO), vitamin E (V(E)) fortified FO (V(E)-FO), V(E) , and PS fortified FO (V(E)-PS-FO), respectively, for 4 wk. The results showed that V(E) fortified FO enhanced the enzymatic (SOD and GPX) and nonenzymatic (GSH and V(E)) antioxidant system, lowered the lipid peroxide (TBARS) concentration compared with FO (P < 0.05). And FO fortified with PS significantly reduced the plasma TG, TC and LDL-C levels, and hepatic TG and TC levels of rats compared with FO (P < 0.05), but had no significant effect on antioxidant defense capacities. Combined addition of V(E) and PS in FO had a synergetic effect. These results indicated that flaxseed oils fortified micronutrients V(E) and PS may contribute to reduce the risk factors of cardiovascular disease (CVD) by improving plasma antioxidant defenses and lipids profiles. Flaxseed oil usually contains greater than 50% of alpha-linolenic acid (ALA) and is a desire origin for n-3 PUFA. But consuming high dose of n-3 PUFA could lead to oxidative damage through free radical-chain reaction in cellular and subcellular membranes. Our studies showed that a regular intake of V(E) and PS fortified flaxseed oils increased antioxidant defenses and ameliorated lipids profile in high-fat fed rats, and these indicated that the flaxseed oil fortified with these micronutrients might reduce the incidence of CVD.


Osteogenic differentiation of 3D-printed porous tantalum with nano-topographic modification for repairing craniofacial bone defects.
Introduction: Congenital or acquired bone defects in the oral and cranio-maxillofacial (OCMF) regions can seriously affect the normal function and facial appearance of patients, and cause great harm to their physical and mental health. To achieve good bone defect repair results, the prosthesis requires good osteogenic ability, appropriate porosity, and precise three-dimensional shape. Tantalum (Ta) has better mechanical properties, osteogenic ability, and microstructure compared to Ti6Al4V, and has become a potential alternative material for bone repair. The bones in the OCMF region have unique shapes, and 3D printing technology is the preferred method for manufacturing personalized prosthesis with complex shapes and structures. The surface characteristics of materials, such as surface morphology, can affect the biological behavior of cells. Among them, nano-topographic surface modification can endow materials with unique surface properties such as wettability and large surface area, enhancing the adhesion of osteoblasts and thereby enhancing their osteogenic ability. Methods: This study used 3D-printed porous tantalum scaffolds, and constructed nano-topographic surface through hydrothermal treatment. Its osteogenic ability was verified through a series of in vitro and in vivo experiments. Results: The porous tantalum modified by nano-topographic surface can promote the proliferation and osteogenic differentiation of BMSCs, and accelerate the formation of new bone in the Angle of the mandible bone defect of rabbits. Discussion: It can be seen that 3D-printed nano-topographic surface modified porous tantalum has broad application prospects in the repair of OCMF bone defects.


Combating malaria in Africa.
The spread of antimalarial drug resistance has major consequences for malaria control in tropical Africa. Here, the impact of chloroquine resistance on the burden of malaria is analyzed and its implications for the Roll Back Malaria initiative are examined. Malaria mortality has increased at least twofold during the past two decades. Combination therapy should be available for home treatment of young children. The potential toxicity of most antimalarials will require special surveillance programs. The main contribution to malaria control using methods to reduce the entomological inoculation rate is expected in areas with low or unstable transmission. Classic vector-control methods could potentially eliminate malaria in most urban areas and such programs deserve high priority.


Durability of branches in branched and fenestrated endografts.
Branched and fenestrated repair has been shown to be effective for treatment of complex aortic aneurysms. However, the long-term durability of branches is not well reported. Prospective data collected for all patients enrolled in a physician-sponsored investigational device exemption trial for branched and fenestrated endografts were analyzed. Retrospective review of imaging studies and electronic records was used to supplement the dataset. Incidences of branch stent secondary intervention, stent fracture, migration, branch-related rupture, and death were calculated. A time-to-event analysis was performed for secondary intervention for any branch. Univariable and multivariable analyses were performed to identify related variables. Branch instability, a composite outcome of any branch event, was reported as a function of exponential decay to capture the loss of freedom from complications over time. Between the years 2001 and 2010, 650 patients underwent endovascular aortic repair with branched or fenestrated devices. Over 9 years of follow-up (mean [standard deviation], 3 [2.3] years), secondary procedures were performed for 0.6% of celiac, 4% of superior mesenteric artery (SMA), 6% of right renal artery, and 5% of left renal artery stents. Mean time to reintervention was 237 (354) days. The 30-day, 1-year, and 5-year freedom from branch intervention was 98% (95% confidence interval [CI], 96%-99%), 94% (95% CI, 92%-96%), and 84% (95% CI, 78%-90%), respectively. Death from branch stent complications occurred in three patients, two related to SMA thrombosis and one due to an unstented SMA scallop. Multivariable analysis revealed no factors as independent predictors of need for branch reintervention. Branches, after branched or fenestrated aortic repair, appear to be durable and are rarely the cause of patient death. The absence of long-term data on branch patency in open repair precludes comparison, yet the lower morbidity and mortality risk coupled with longer-term durability data will further alter the balance of repair options.


In vitro metabolism of the analgesic bicifadine in the mouse, rat, monkey, and human.
The in vitro metabolism of [(14)C]bicifadine by hepatic microsomes and hepatocytes from mouse, rat, monkey, and human was compared using radiometric high-performance liquid chromatography and liquid chromatography/tandem mass spectrometry. Two main metabolic pathways were identified in all four species. One pathway was an NADPH-dependent pathway in which the methyl group was oxidized to form a hydroxymethyl metabolite (M2). Its formation was inhibited in human microsomes only by quinidine, a CYP2D6 inhibitor. In incubations with individual cDNA-expressed human cytochromes P450, M2 was formed only by CYP2D6 and CYP1A2, with CYP2D6 activity 6-fold greater than that of CYP1A2. M2 was oxidized further to the carboxylic acid metabolite (M3) by hepatocytes from all four species. The second major metabolic pathway was an NADPH-independent oxidation at the C2 position of the pyrrolidine ring, forming a lactam metabolite (M12). This reaction was almost completely inhibited in human hepatic microsomes and mitochondria by the monoamine oxidase (MAO)-B-specific inhibitor selegiline. Clorgyline, a specific inhibitor of MAO-A, was less effective in inhibiting M12 formation. Other metabolic pathways of variable significance among the four species included the formation of carbamoyl-O-glucuronide, hydroxymethyl lactam, and carboxyl lactam. Overall, the data indicate that the primary enzymes responsible for the primary metabolism of bicifadine in humans are MAO-B and CYP2D6.


Organising the MRCP exam--a guide for registrars.
Taking the MRCP exam as a candidate can be traumatic. For the registrar who has to organise one, the task may be just as daunting. Efficient coordination of candidates, patients and examiners and the provision of good clinical material in a suitable setting requires considerable organisation. This supplement to the guide issued by the College, based on our own experience, describes in more detail the preparation and management of the examination in the hope that it will ease our colleagues' tasks.


IGF-II expression and methylation in small for gestational age infants.
Low birth weight is associated with an increased risk of adverse outcomes in many diseases in adult life. We investigated the expression of IGF-II and the status of differentially methylated regions (DMR) in small for gestational age (SGA) infants after birth. Plasma IGF-II, IGF-II receptor (IGF2R), IGF-I, and IGF-binding protein 3 (IGFBP3) levels were measured after birth in 150 newborn infants. These included 30 term appropriate for gestational age (AGA), 30 term SGA, 30 term large for gestational age (LGA), 30 preterm AGA, and 30 preterm SGA infants. H19 and IGF2 mRNA levels were quantified by fluorescence quantitative polymerase chain reaction (PCR). The methylation status of H19 DMR and IGF2 DMR2 was assessed by methylation-specific PCR (MSP). Plasma IGF-II, IGF2R, IGF-I, and IGFBP3 levels were measured by enzyme-linked immunosorbent assay in AGA infants of different gestational ages and term AGA infants on different days. Plasma IGF-II levels after birth were lower in both term SGA (435.1±33.82 vs. 620.4±44.79, p=0.002) and LGA infants (483.7±33.8 vs. 620.42±44.79, p=0.018) than in term AGA infants. The expression of IGF2 mRNA was higher in the term SGA group than in the preterm SGA group (p=0.014) and the preterm SGA group displayed lower expression than the preterm AGA group (p=0.048). The H19 DMR methylation level was higher in both term SGA (p=0.044) and term LGA infants (p=0.027) compared to term AGA infants. IGF-II was associated with birth weight and expressed at high levels, which suggests that IGF-II may continue to play an important role after birth. H19 DMR methylation may be involved in controlling IGF-II expression.


Incorrect key presses may cause Nutricia Flocare Infinity series enteral feeding pumps to appear to be infusing even though an occlusion exists.
If users of Nutricia Flocare Infinity or Flocare Infinity+ feeding pumps press the unit's Fill Set key while attempting to address an occlusion alarm, the unit's display may indicate normal device operation even though the unit is not delivering feeding solution. Failure to deliver solution may result in hypoglycemia, which can have severe outcomes for patients who have difficulty maintaining their blood sugar levels. Flocare users should become familiar with the issue and with the guidance offered in the products' instructions for use.


Magnetic resonance imaging of perilymphatic fistula.
Perilymphatic fistula (PLF) is considered to be a most challenging otologic issue. There are no currently agreed upon objective tests for the diagnosis of PLF. In an effort to improve diagnostic accuracy in patients with suspected PLF, a novel diagnostic test involving magnetic resonance (MR) contrast imaging was designed. An experimental PLF was created in the cochlear round window membrane of healthy adult cats. Since cochlear perilymph is thought to be an ultrafiltrate of cerebral spinal fluid, gadodiamide (gadolinium-diethylenetriamine penta-acetic acid [DTPA] bismethylamide), a nonionic paramagnetic contrast agent, was injected intrathecally in an attempt to enhance imaging of the created fistula. Post-contrast images of the fistualized cochlea demonstrate a significant increase in the signal intensity of the cochlear perilymph with pooling of enhanced perilymph observed in the ipsilateral mastoid bulla. Magnetic resonance contrast imaging may prove to be a valuable technique in human studies involving perilymphatic fistula.


Asymmetric autocatalysis of pyrimidyl alkanol and its application to the study on the origin of homochirality.
CONSPECTUS: Amplification of enantiomeric excess (ee) is a key feature for the chemical evolution of biological homochirality from the origin of chirality. We describe the amplification of ee in the asymmetric autocatalysis of 5-pyrimidyl alkanols in the reaction between diisopropylzinc (i-Pr2Zn) and pyrimidine-5-carbaldehydes. During the reaction, an extremely low ee (ca. 0.00005% ee) can be amplified to >99.5% ee, and therefore, the initial slightly major enantiomer is automultiplied by a factor of ca. 630000, while the initial slightly minor enantiomer is automultiplied by a factor of less than 1000. In addition, pyrimidyl alkanols with various substituents at the 2-position of the pyrimidine ring, 3-quinolyl alkanol, 5-carbamoyl-3-pyridyl alkanol, and large multifunctionalized pyrimidyl alkanols also act as highly efficient asymmetric autocatalysts in the addition of i-Pr2Zn to the corresponding aldehydes. The asymmetric autocatalysis of pyrimidyl alkanol can discriminate the chirality of various compounds. Chiral substances such as alcohols, amino acids, hydrocarbons, metal complexes, and heterogeneous chiral materials can act as chiral triggers for asymmetric autocatalysis to afford pyrimidyl alkanols with the corresponding absolute configuration of the initiator. This recognition ability of chiral compounds is extremely high, and chiral discrimination of a cryptochiral quaternary saturated hydrocarbon was established by applying asymmetric autocatalysis. By using the large amplification effect of the asymmetric autocatalysis, we can link various proposed origins of chirality with highly enantioenriched organic compounds in conjunction with asymmetric autocatalysis. Thus, a statistical fluctuation in ee of racemic compounds can be amplified to high ee by using asymmetric autocatalysis. Enantiomeric imbalance induced by irradiation of circularly polarized light can affect the enantioselectivity of asymmetric autocatalysis. The asymmetric autocatalysis was also triggered by the morphology of inorganic chiral crystals such as quartz, sodium chlorate, and cinnabar. Chiral organic crystals of achiral compounds also act as chiral initiators, and during the study of a crystal of cytosine, enantioselective chiral crystal phase transformation of the cytosine crystal was achieved by removal of the water of crystallization in an achiral monohydrate crystal. Enantioselective C-C bond formation was realized on the surfaces of achiral single crystals based on the oriented prochirality of achiral aldehydes. Furthermore, asymmetric autocatalysis of pyrimidyl alkanols is a highly sensitive reaction that can recognize and amplify the significantly small effect of a chiral compound arising solely from isotope substitution of hydrogen, carbon, and oxygen (D/H, (13)C/(12)C, and (18)O/(16)O). These examples show that asymmetric autocatalysis with an amplification of chirality is a powerful tool for correlating the origin of chirality with highly enantioenriched organic compounds. Asymmetric autocatalysis using two β-amino alcohols reveals a reversal of enantioselectivity in the addition of i-Pr2Zn to aldehyde and is one approach toward understanding the mechanism of asymmetric dialkylzinc addition, where heteroaggregates act as the catalytic species.


Higher Dialysate Matrix Metalloproteinase-2 Levels Are Associated with Peritoneal Membrane Dysfunction.
♦ Peritoneal dialysis (PD) patients develop progressive and cumulative peritoneal injury with longer time spent on PD. The present study aimed to a) describe the trend of peritoneal injury biomarkers, matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1), in incident PD patients, b) to explore the capacity of dialysate MMP-2 to predict peritoneal solute transport rate (PSTR) and peritonitis, and c) to evaluate the influence of neutral pH, low glucose degradation product (GDP) PD solution on these outcomes. ♦ The study included 178 participants from the balANZ trial who had at least 1 stored dialysate sample. Changes in PSTR and peritonitis were primary outcome measures, and the utility of MMP-2 in predicting these outcomes was analyzed using multilevel linear regression and multilevel Poisson regression, respectively. ♦ Significant linear increases in dialysate MMP-2 and TIMP-1 concentrations were observed (p < 0.001), but neither was affected by the type of PD solutions received (MMP-2: p = 0.07; TIMP-1: p = 0.63). An increase in PSTR from baseline was associated with higher levels of MMP-2 (p = 0.02), and the use of standard solutions over longer PD duration (p = 0.001). The risk of peritonitis was independently predicted by higher dialysate MMP-2 levels (incidence rate ratio [IRR] per ng/mL 1.01, 95% confidence interval [CI] 1.005 - 1.02, p = 0.002) and use of standard solutions (Biocompatible solution: IRR 0.45, 95% CI 0.24 - 0.85, p = 0.01). ♦ Dialysate MMP-2 and TIMP-1 concentrations increased with longer PD duration. Higher MMP-2 levels were associated with faster PSTR and future peritonitis risk. Administration of biocompatible solutions exerted no significant effect on dialysate levels of MMP-2 or TIMP-1, but did counteract the increase in PSTR and the risk of peritonitis associated with the use of standard PD solutions. This is the first longitudinal study to examine the clinical utility of MMP-2 as a predictor of patient-level outcomes.


Chemical modification of siRNA.
siRNAs are widely used for the control of gene expression in molecular biology and experimental pharmacology. siRNA molecules are the most efficient and specific inhibitors of gene expression to have been discovered, acting at nanomolar and subnanomolar concentrations. Challenges associated with nuclease stability, selecting effective sequences, reducing off-target effects and achieving efficient delivery to target cells and tissues have significantly limited the biomedical applications of siRNA. However, chemical modification is a powerful tool for improving the properties of siRNA. This review examines recent data regarding chemical modifications used to develop potent effectors of gene silencing. The correlation between thermal stability and potency, and between nuclease resistance and duration of the silencing effect, are discussed with relation to chemically modified siRNAs.


[Overcoming of multidrug resistance by a newly synthesized quinoline compound, MS-209].
The emergence of multidrug resistance(MDR) is a major problem in cancer chemotherapy. Many compounds are developed to reverse MDR, and some of them are under clinical trials. Among them, MS-209, a novel quinoline derivative, is one of the most potent MDR reversing agents. MS-209 at 3 microM effectively reverses MDR in various cell lines in vitro. MS-209 directly interacts with P-glycoprotein and inhibits the P-glycoprotein-mediated drug transport. Oral administration of MS-209 combined with anticancer drugs markedly increases the life span of mice bearing MDR tumor cells without causing serious side effects. Thus, MS-209 is an orally active and potent MDR reversing drug without serious side effects.


Real-time atomic force microscopy using mechanical resonator type scanner.
The real-time atomic force microscope for biological sample is a challenging research field. We have demonstrated a real-time atomic force microscope by implementing a mechanical resonator type scanner called by "microscanner" The microscanner was designed to have a resonance frequency in the range of 5-10 kHz and an amplitude of 1-3 microm. The resonant vibration of the microscanner was served as a fast-scan directional motion, and an image acquisition rate of 30 frames/s with 256x256 pixels per frame was achieved. Time-varying sequential images of a poly(ethylene-oxide) sample were taken as a demonstration of potential for excellence in real-time imaging a moving nano-object.


Cyclophanes as Platforms for Reactive Multimetallic Complexes.
Multimetallic cofactors supported by weak-field donors frequently function as reaction centers in metalloproteins, and many of these cofactors catalyze small molecule activation (e.g., N2, O2, CO2) with prominent roles in geochemical element cycles or detoxification. Notable examples include the iron-molybdenum cofactor of the molybdenum-dependent nitrogenases, which catalyze N2 fixation, and the NiFe4S4 cluster and the Mo(O)SCu site in various carbon monoxide dehydrogenases. The prevailing proposed reaction mechanisms for these multimetallic cofactors relies on a cooperative pathway, in which the oxidation state changes are distributed over the aggregate coupled with orbital overlap between the substrate and more than one metal ion within the cluster. Such cooperativity has also been proposed for chemical transformations at the surfaces of heterogeneous catalysts. However, the design details that afford cooperative effects and allow such reactivity to be harnessed effectively in homogeneous synthetic systems remain unclear. Relatedly, hydride donors ligated to these metal cluster cofactors are suggested as precursors to the state that reacts with substrates; here too, however, the reactivity of hydride-decorated clusters supported by weak-field ligands is underexplored. Inspired by the reactivity potential of multimetallic assemblies evidenced in biological systems, approaches to design, synthesize, and evaluate reactivity of polynuclear metal compounds have been actively explored. In a similar vein to the templating function afforded by enzyme active sites, a carefully engineered organic ligand can be employed to control metal nuclearity of the complex and the local coordination environment of each metal center. This Account presents our efforts within this field, beginning with ligand design considerations followed by a survey of observed small molecule activation by trimetallic cyclophanates. We highlight the distinct reactivity outcomes accessed by multimetallic compounds as compared to aggregates that assemble in reaction mixtures from monometallic precursors. Contributing to the opportunity for programmed cooperativity in these designed multimetallic compounds, the cyclophane also dictates the orientation of substrate binding and metal-substrate interactions, which has a prominent influence on reactivity. For example, the dinitrogen-tricopper(I) cyclophanate reacts with dioxygen with markedly different results as compared to monocopper compounds. As an unexpected outcome, one series of tricopper compounds were discovered to be competent catalysts for carbon dioxide reduction to oxalate-a formally one-electron process-hinting at an inherently broader reaction scope for weak-field clusters at lowering the barrier for one-electron pathways as well as multielectron redox transformations. Further reflecting the role of the ligand in tuning reactivity, the trimetallic trihydride cluster compounds, [M3(μ-H)3]3+ (M = FeII, CoII, ZnII), demonstrate substrate specificity for CO2 over various other unsaturated molecules and surprising stability toward water. This series reflects the role of the local environment of a shallow ligand pocket to control substrate access. Summed together, the systems described here evidence the anticipated cooperative reactivity accessed in designed multimetallic species vs self-assembled monometallic systems (e.g., O2 activation and O atom transfer) as well as control of substrate access by seemingly subtle structural effects. Indeed, future efforts aim to interrogate the limits of cooperativity in these systems as well as the role of ligand dynamics and sterics on reactivity.


Use of food intake and body mass index (BMI) in the assessment of adult nutritional status in Viet Nam including a maternal-child analysis.
The authors have used food intake and Body Mass Index (BMI) to assess Vietnamese adult nutritional status. Chronic Energy Deficiency (CED) is prevalent in Vietnamese adults with an average percentage about 40%. The mean value of BMI in the 26-40 year old age group is 19.7 but it decreases thereafter except in urban areas. The change in the BMI curve distribution varies among adults living in rural, urban and mountainous areas. There seems to be a relationship between the BMI of mothers and the nutritional status of their children underthe age of 5. Some findings revealed a relationship between maternal BMI and birth weight and between CED and health status. The proposed cut-off point of Ferro-Luzzi-James in the classification of CED was applied to data from Vietnam.


Secretome analysis of human BMSCs and identification of SMOC1 as an important ECM protein in osteoblast differentiation.
Extracellular matrix proteins have been implicated in the regulation of osteoblast differentiation of bone marrow derived mesenchymal stem cells (BMSCs) through paracrine or autocrine mechanisms. In the current study, we analyzed the secretory protein profiles of BMSCs grown in osteogenic medium (OSM) and identified SPARC-related modular calcium-binding protein 1 (SMOC1), a member of the SPARC family, as a regulator of osteoblast differentiation of BMSCs. BMSCs with high and low osteogenic potential were grouped and stimulated with OSM, after which conditioned medium was collected and analyzed by LC-MS/MS. We identified 410 proteins, 64 of which were selectively secreted by high osteogenic potential BMSCs. Of these 64 secreted proteins, we selected extracellular matrix proteins for validation in BMSCs undergoing osteoblast differentiation and found that SMOC1 is highly expressed and secreted in BMSCs stimulated with OSM. To examine the role of SMOC1 in osteoblast differentiation, we analyzed the effect of SMOC1 knockdown and overexpression using shRNAs and wild-type cDNA, respectively. Knockdown of SMOC1 significantly inhibited mineralization and the expression of osteoblast differentiation markers, while overexpression of SMOC1 substantially increased the expression of osteoblast differentiation-related genes. Thus, validation of secretome profiling data identified SMOC1 as a putative regulator of osteoblast differentiation of BMSCs.


High resolution fast quantitative docking using Fourier domain correlation techniques.
A 'docking' method based on finite grid forcefield sampling is proposed for fast evaluation of interaction energies between macromolecules and ligands. Forcefield used to calculate interaction energies utilizes a potential energy function composed of a 1/r-dependent electrostatic term and a (6-12) Lennard-Jones term for van der Waals interactions. Fast evaluation makes use of the convolution theorem allowing a point-by-point N-dimensional correlation in direct space to be replaced by a simple multiplication in spatial frequency space. Predictive accuracy was assessed by using seven protein-ligand complexes available from the Brookhaven Data Bank and determined crystallographically to high resolution. Successful prediction of ligand position and determination of ligand-protein interaction enthalpy was dependent on forecefield sampling grid size. Minimum interaction enthalpy calculated for four protein-ligand complexes coincided with crystallographic structures that used sampling grid sizes of 0.25 A resolution and was independent of ligand starting position and orientation. Successful docking was obtained for the remaining complexes at same grid resolution provided ligand starting positions were not randomized. Sensitivity of the docking algorithm to starting orientation was a consequence of tight fit of respective ligand structures with their protein target sites for these three cases and can be circumvented by using finer rotational sampling grids for the ligand. Boltzmann statistics derived from calculated interaction energies successfully extracted the observed ribonuclease A cytidylic acid complex from a manifold of similar interaction energies. The proposed method was able to reproduce the observed crystallographic complex by using a dynamical description of ligand.


Functional outcome of orthotopic bladder substitution: a comparison between the S-shaped and U-shaped neobladder.
To compare the functional outcome of two types of orthotopic bladder substitution, the S-shaped and the U-shaped neobladder, with respect to leakage, functional capacity and quality of life. Between 1999 and 2007, 45 male patients with urinary bladder cancer were treated with cystectomy and orthotopic bladder substitution; 23 with the S-shaped bladder ad modum Schreiter and 22 with the U-shaped bladder ad modum Studer. Patients were followed up by a urologist and a specialized nurse (urotherapist) at 1, 3 and 6 months. At each visit the patient completed a voiding chart, a weighted pad test and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30). Differences between the groups in functional outcome and quality of life variables were analysed by Student's t test using SPSS software. Mean maximum bladder capacity increased over time. At 6 months, the S-bladder had a larger capacity than the U-bladder (525 ml vs 423 ml). Patients with an S-bladder had less urine leakage at all follow-ups, although this was statistically significant only at 6 months regarding day-time incontinence and at all visits regarding night-time incontinence. The mean urine leakage at 6 months was 7 g (day) and 30 g (night) in the S-bladder group and 50 g (day) and 250 g (night) in the U-bladder group. However, quality of life did not differ between the groups. The S-bladder had better bladder capacity and less leakage than the U-bladder, but these differences did not translate into differences in quality of life. The results should be confirmed in larger prospective studies.


6-0 nylon versus 6-0 vicryl rapide in chieloplasty.
Facial cosmetic result is one of the most concerning issues for the parents who get their children operated for cleft lip. Moreover, the discomfort associated with the suture removal encourages one to use any new technology that may replace the need for suture placement. The type of suture material used in surgery has been a long-standing debate among surgeons. In this study, we compared rapidly absorbable suture material (Vicryl Rapide™) with nonabsorbable suture material (nylon). The aim of this study is to compare the appearance and course of scar, wound infection, and patient's parent perception using Vicryl Rapide and nylon in nonsyndromic congenital cleft lip repair. This was a randomized prospective controlled clinical trial. Twenty patients, in the age group of 3-18 months treated for unilateral congenital cleft lip deformity, were included and randomly allocated to two groups with ten patients each. Skin suturing was done with 6-0 polyamide and 6-0 irradiated polyglactin in Groups A and B, respectively. Patients were evaluated at 1 week, 1, 3, 6 months, and 1 year postoperatively in person by the observer as well as by the patient's parent. Descriptive statistical analysis was done using SPSS 20, and Student's t-test was applied. It was found that Vicryl Rapide showed more hypopigmented scars and raised scars than nylon at the end of 1 year though overall appearance was comparable between the groups. Vicryl Rapide showed poorer cosmetic outcomes in terms of height and pigmentation of car as compared to nylon suture of same thickness. However, since scars tend to improve with time, a bigger sample size and a longer follow-up are required to generalize this statement.


Gegen Qinlian Decoction Attenuates High-Fat Diet-Induced Steatohepatitis in Rats via Gut Microbiota.
Gut microbiota play an important role in modulating energy contribution, metabolism, and inflammation, and disruption of the microbiome population is closely associated with chronic metabolic diseases, such as nonalcoholic fatty liver disease (NAFLD). Gegen Qinlian decoction (GGQLD), a well-known traditional Chinese herbal medicine (CHM), was previously found to regulate lipid metabolism and attenuate inflammation during NAFLD pathogenesis. However, the underlying mechanism of this process, as well as how the gut microbiome is involved, remains largely unknown. In this study, we investigated the effect of varying doses of GGQLD on the total amount and distribution of gut bacteria in rats fed a high-fat diet (HFD) for 8 weeks. Our analysis indicates that Oscillibacter and Ruminococcaceae_g_unclassified are the dominant families in the HFD group. Further, HFD-dependent differences at the phylum, class, and genus levels appear to lead to dysbiosis, characterized by an increase in the Firmicutes/Bacteroidetes ratio and a dramatic increase in the Oscillibacter genus compared to the control group. Treatment with GGQLD, especially the GGQLL dose, improved these HFD-induced changes in intestinal flora, leading to increased levels of Firmicutes, Clostridia, Lactobacillus, bacilli, and Erysipelotrichales that were similar to the controls. Taken together, our data highlight the efficacy of GGQLD in treating NAFLD and support its clinical use as a treatment for NAFLD/NASH patients.


Association of variants in SH2B1 and RABEP1 with worsening of low-density lipoprotein and glucose parameters in patients treated with psychotropic drugs.
Genetic factors associated with Body Mass Index (BMI) have been widely studied over the last decade. We examined whether genetic variants previously associated with BMI in the general population are associated with cardiometabolic parameter worsening in the psychiatric population receiving psychotropic drugs, a high-risk group for metabolic disturbances. Classification And Regression Trees (CARTs) were used as a tool capable of describing hierarchical associations, to pinpoint genetic variants best predicting worsening of cardiometabolic parameters (i.e total, HDL and LDL-cholesterol, triglycerides, body mass index, waist circumference, fasting glucose, and blood pressure) following prescription of psychotropic drugs inducing weight gain in a discovery sample of 357 Caucasian patients. Significant findings were tested for replication in a second Caucasian psychiatric sample (n=140). SH2B1 rs3888190C>A was significantly associated with LDL levels in the discovery and in the replication sample, with A-allele carriers having 0.2mmol/l (p=0.005) and 0.36mmol/l (p=0.007) higher LDL levels compared to others, respectively. G-allele carriers of RABEP1 rs1000940A>G had lower fasting glucose levels compared to others in both samples (-0.16mmol/l; p<0.001 and -0.77mmol/l; p=0.03 respectively). The present study is the first to observe such associations in human subjects, which may in part be explained by a high risk towards dyslipidemia and diabetes in psychiatric patients receiving psychotropic treatments compared to population-based individuals. These results may therefore give new insight into the etiology of LDL-cholesterol and glucose regulation in psychiatric patients under psychotropic drug therapy.


May furtive predation provide enemy free space in ant-tended aphid colonies?
In furtive predation, a predator is able to exploit its prey without generating significant defensive behaviors from them. However, in aphidophagous guild, if furtive predator can benefit from dilution effects generated by the aphids, they also suffer from intraguild predation from more mobile and active-searching predators. In this context ant-tended aphid colonies might not only represent an important food source but also potentially an enemy-free space for furtive predators if they remain unharmed by ants while other active predators are being repelled. Here we use the furtive predator Aphidoletes aphidimyza and two distinct instars of an active-searching predator, the Asian ladybeetle Harmonia axyridis, to test hypotheses related to predator persistence within aphid colonies in presence of ants. Our results show that persistence rate over time of the furtive predator was not affected by ant presence while it was strongly reduced for both instars of the active-searching predator. Furthermore, when ran in paired trials within ant-tended aphid colonies, furtive predator persistence rate was significantly higher than for active-searching predators, with these latter always leaving the plants quicker. Finally, we tested the importance of predator mobility in detection susceptibility and aggressive responses in ants using mobile and immobile active-searching predators. While the number of antennal palpations was similar for both treatments indicating similar detection rate, the number of ant attacks was significantly higher on mobile individuals highlighting the importance of movement in triggering aggressive responses in ants. Overall our results indicate that furtive predation represents an efficient strategy to limit ant aggressions, while the exclusion of active-searching predators might create an enemy-free space for furtive predators within ant-tended aphid colonies.


In vitro glucuronidation of thyroxine and triiodothyronine by liver microsomes and recombinant human UDP-glucuronosyltransferases.
Glucuronidation, which may take place on the phenolic hydroxyl and carboxyl groups, is a major pathway of metabolism for thyroxine (T4) and triiodothyronine (T3). In this study, a liquid chromatography/mass spectrometry (LC/MS) method was developed to separate phenolic and acyl glucuronides of T4 and T3. The method was used to collect the phenolic glucuronide of T4 for definitive characterization by NMR and to determine effects of incubation pH, species differences, and human UDP-glucuronosyltransferases (UGTs) involved in the formation of the glucuronides. Formation of T4 phenolic glucuronide was favored at pH 7.4, whereas formation of T4 acyl glucuronide was favored at pH 6.8. All the UGTs examined catalyzed the formation of T4 phenolic glucuronide except UGT1A4; the highest activity was detected with UGT1A3, UGT1A8, and UGT1A10, followed by UGT1A1 and UGT2B4. Formation of T3 phenolic glucuronide was observed in the order of UGT1A8 > UGT1A10 > UGT1A3 > UGT1A1; trace activity was observed with UGT1A6 and UGT1A9. UGT1A3 was the major isoform catalyzing the formation of T4 and T3 acyl glucuronides. In liver microsomes, phenolic glucuronidation was the highest in mice for T4 and in rats for T3 and lowest in monkeys for both T4 and T3. Acyl glucuronidation was highest in humans and lowest in mice for T4 and T3. Phenolic glucuronidation was higher than acyl glucuronidation for T4 in humans; in contrast, the acyl glucuronidation was slightly higher than phenolic glucuronidation for T3. UGT activities were lower toward T3 than T4 in all the species. The LC/MS method was a useful tool in studying glucuronidation of T4 and T3.


[Leiomyosarcoma of the small intestine: a rare cause of gastrointestinal blood loss].
A 61-year old man is described with recurrent gastrointestinal bleeding, due to a metastatic jejunal leiomyosarcoma. Because of the low incidence and the aspecific symptoms, a long delay occurred until the bleeding tumour and its metastases were demonstrated at radionuclide scanning and mesenteric angiography. The tumour-bearing segment could be removed by simple surgical excision.


Laparoscopic suture rectopexy for rectal prolapse in a 22-month-old child.
We present the case of a 22-month-old female child who presented with severe recurrent rectum prolapse. The patient was successfully managed using the laparoscopic simple suture rectopexy approach with 5-mm instruments employing two 3-0 nonabsorbable sutures on either side of the rectum to secure it to the presacral fascia. There was no blood loss, and the procedure was completed without complication. The child was followed up for a period of 24 months with good results.


Diagnosis and Control of a LPAI H5N8 Outbreak in a Japanese Quail (Coturnix coturnix japonica) Commercial Flock in the Central Valley of California.
In April 2014 an outbreak of low pathogenic avian influenza H5N8 North American genetic lineage was diagnosed in a commercial quail operation in Stanislaus County, California. Sudden increase in mortality prompted the submission of 20 Japanese quail hens (Coturnix c. japonica) to the California Animal Health and Food Safety Laboratory, Turlock Branch. Oropharyngeal and cloacal swabs tested positive for influenza A virus H5N8 by real-time reverse transcription-polymerase chain reaction. The virus was subsequently isolated. In vivo assay and sequencing of the hemagglutinin protein cleavage site classified the virus as a North American genetic lineage of low pathogenicity for chickens. Following the diagnosis, a rapid and coordinated response took place to contain the outbreak. The affected premise was depopulated, cleaned, and disinfected. Three areas from the affected premises-a 3 kilometer (km) radius (High Risk Zone), a 3-10 km area (Buffer Zone), and a 10-20 km (Surveillance Zone)-were established for avian influenza testing of commercial and noncommercial poultry operations. Surveillance testing and rapid control measures were successful in the control and eradication of the outbreak and revealed no area of spread of the virus from the index flock. This report describes the history, diagnosis, surveillance, and control measures applied to manage this outbreak.


Effect of magnesium sulfate on the calcium-stimulated adenosine triphosphatase activity and lipid peroxidation of red blood cell membranes from preeclamptic women.
The effect of the treatment with magnesium sulfate (MgSO(4)) on Ca-ATPase activity and level of lipid peroxidation of red blood cells from preeclamptic pregnant women was examined because it is known that these parameters are affected with preeclampsia. Red cell ghosts from 11 normotensive and 11 preeclamptic pregnant women, before and after treatment with MgSO(4), were assayed for Ca-ATPase activity and level of lipid peroxidation, determined as TBARS or conjugated dienes. It was found that the Ca-ATPase activity is significantly lower and the level of lipid peroxidation is significantly higher in the preeclamptic women with no treatment, as compared to normotensive pregnant women. Both parameters return to normal values after the MgSO(4) therapy. These results can be mimicked by in vitro preincubation with MgSO(4) of intact red blood cells from preeclamptic pregnant women, without any treatment. Our data indicate that MgSO(4) treatment of preeclamptic pregnant women modifies both the Ca-ATPase activity and the level of lipid peroxidation of their red blood cell membranes, reaching values similar to those of normotensive pregnant women. The diminution of the level of lipid peroxidation by MgSO(4), can account for the increase in Ca-ATPase activity.


Studies on vesicourethral reflexes. II. Urethral sphincteric responses to spinal cord stimulation.
A bipartite bladder model was developed with and without complete muscular separation in order to isolate and analyze separately sphincteric and detrusor responses. Spinal cord stimulation of the sacral micturition center evoked responses in the detrusor compartment as well as in the urethral sphincteric compartment. The micturition center was easily definable in the spinal cord at a variable point lying in the segment L51/2 to L6 vertebral levels. It was not possible to isolate a separate detrusor center from a sphincteric center. They did overlap-with the sphincteric center extending both above and below the detrusor center. Parameters of stimulation were tested and it was noted that 1 to 3 v, frequency of 10 to 15 cps, and duration of 1 msec gave the optimal responses. Complte muscular separation did not alter sphincteric response to spinal cord stimulation. Nerve-mediated impulses resulted in rise in detrusor pressure and a simultaneous rise in urethral pressure that interfered with proper voiding and hindered bladder emptying. This problem remains to be solved before central spinal cord stimulation for the purpose of controlled bladder evacuation becomes fully effective.


Seasonal variations in the food intake of captive red grouse.
1. The daily food intake of captive red grouse was measured throughout the year. It was positively related to body weight and to daylength, and negatively to air temperature. Grouse ate more during their main summer moult, and the hens ate most while producing eggs. 2. The total weight of eggs laid was positively related to daily food intake during the 5 weeks before laying, but not to daily intake during laying. The rate at which eggs were laid was related to daily intake during laying as well as before laying. The possible significance of these results for poultry management is discussed.


Immunoreactivity evaluation of mutant p53 gene product with DNA ploidy pattern in colorectal carcinoma.
Studying p53 protein expression in tumor cells is one of the effective methods for detecting p53 gene mutations. This study attempted simultaneous monitoring of p53 overexpression in colon cancer using immunohistochemical and immunoblotting techniques and also to compare abnormalities of p53 with DNA ploidy and clinicopathological variables. The occurrence of p53 protein expression was analyzed in forty-nine fresh colorectal cancer specimens by immunohistochemical and p53 protein expression also demonstrated by Western immunoblotting technique in 28 colorectal cancer specimens, using an anti-human p53 monoclonal antibody (Do-7), and 25 normal colon mucosa as a negative control. DNA ploidy in 36 specimens of colon cancer tissues was determined by Flow cytometry. Overexpression of p53 protein was detected immunohistochemically in 53.1% (26 of 49) of the tumor specimens. DNA ploidy was performed in 36 cases, 55.6% (20 of 36) of colon cancer specimens were DNA aneuploidy, p53 immunostaining was positive in 60% of cases with DNA aneuploidy compared to 31.3% in diploid tumors (p<0.001). There was no significant association between p53 immunostaining and clinicopathological variables. Overexpression of p53 protein was demonstrated in nuclear protein extract by immunoblotting in 75% (21 of 28) of colorectal carcinoma. Aneuploidy carcinomas were more frequently p53 positive by immunoblotting than DNA diploidy carcinomas; 76.5% (13 of 17) vs. 72.7% (8 of 11) (p<0.2). P53 expression by immunoblotting was more frequently found in good lymphocytic infiltration than moderate and poor lymphocytic infiltration (p<0.001). Also, p53 expression in right colon was significant with rectum (p<0.009). The incidence of p53 expression in Duke's stage B was significant if compared with Duke's stage C (p<0.005). Immuno-reactivity of p53 expression was detected by immunostaining and immunoblotting in 89.3% (25 of 28) of colorectal cancer. P53 immunoreactivity by immunostaining and immunoblotting were closely related to the clinicopathological variables such as pathological type (p<0.01), lymphocytic infiltration (p<0.0001), tumor grade, and tumor site (p<0.001). DNA aneuploidy was more frequently p53 positive than DNA diploid tumor by immunostaining and immunoblotting (p<0.001). Immunohistochemistry confirmed by immunoblotting assay is a sensitive method for detecting the trace amount of p53 protein and provides valuable information for the understanding of colorectal cancer biology.


In resource-limited settings good early outcomes can be achieved in children using adult fixed-dose combination antiretroviral therapy.
To (a) determine early treatment outcomes and (b) assess safety in children treated with adult fixed-dose combination (FDC) antiretroviral tablets. Sixteen Medecins Sans Frontieres (MSF) HIV programs in eight countries in resource-limited settings (RLS). Analysis of routine program data gathered June 2001 to March 2005. A total of 1184 children [median age, 7 years; inter-quartile range (IQR), 4.6-9.3] were treated with antiretroviral therapy (ART) of whom 616(52%) were male. At ART initiation, Centres for Disease Control stages N, A, B and C were 9, 14, 38 and 39%, respectively. Children were followed up for a median period of 6 months (IQR, 2-12 months). At 12 months the median CD4 percentage gain in children aged 18-59 months was 15% (IQR, 6-18%), and the percentage with CD4 gain < 15% was reduced from 85% at baseline to 11%. In those aged 60-156 months, median CD4 cell count gain was 275 cells/microl (IQR, 84-518 cells/microl), and the percentage with CD4 < 200 cells/mul reduced from 51% at baseline to 11%. Treatment outcomes included; 1012 (85%) alive and on ART, 36 (3%) deaths, 15 (1%) stopped ART, 89 (8%) lost to follow-up, and 31 (3%) with unknown outcomes. Overall probability of survival at 12 months was 0.87 (0.84-0.89). Side effects caused a change to alternative antiretroviral drugs in 26 (2%) but no deaths. Very satisfactory early outcomes can be achieved in children in RLS using generic adult FDC antiretroviral tablets. These findings strongly favour their use as an "interim solution" for scaling-up ART in children; however, more appropriate pediatric antiretroviral drugs remain urgently needed.


Improving anti-infrared radiation and heat insulation by potassium hexatitanate whisker-doped Al2O3-SiO2 composite xerogel.
Taking potassium hexatitanate whisker (PTW) modified by silane coupling agent KH550, aluminium nitrate inorganic salt and tetraethyl orthosilicate and deionized water, respectively, as infrared sunscreen, aluminium source and raw materials forming the network structure of a composite xerogel, a series of structurally intact PTW-doped Al2O3-SiO2 composite xerogel thermal-insulating materials were prepared by the sol-gel method and dried under atmospheric pressure. The infrared spectral characteristics, infrared radiation transmittance, microstructures, morphology and thermal conductivity of different composite materials prepared have been determined by a Fourier transform infrared spectrometer, UV-visible-near-infrared spectrophotometer, X-ray diffractometer, scanning electron microscope and thermal conductivity tester. The results exhibit that when the Al to Si molar ratio is 1 : 9, the composite material with 5 wt% modified PTW shows the best infrared radiation blocking performance and the lowest thermal conductivity (0.0604 W m-1 K-1).


[Preparation and evaluation of 2,6-di-O-pentyl-beta-cyclodextrin bonded silica stationary phase for high performance liquid chromatography].
In order to improve the chiral separation capability of the conventional beta-cyclodextrin bonded-silica gel stationary phase, 2,6-di-O-pentyl-beta-cyclodextrin bonded stationary phase (PCDS) was prepared via a long spacer. The resulted bonded-silica stationary phase was characterized by three methods, namely Fourier transform infrared, Molisch color reaction, X-ray optical electrical energy spectrogram. The chromatographic performances of PCDS were investigated by using liquid chromatography with toluene, dimethyl phthalate, and phenanthrene as solutes, and their retention mechanism was investigated and discussed. The results show that the introduction of pentyl to beta-cyclodextrin leads to enhancement of the retention of the solutes. The chiral separation capability of the new bonded-silica stationary phase was evaluated by using liquid chromatography with some chiral drugs. Some of the enantiomers such as chlorphenamine maleate and bupropion hydrochloride were separated by heptakis (2,6-di-O-pentyl)-beta-cyclodextrin bonded silica stationary.


Clinical and Pathologic Response to Vedolizumab in a Young Female Patient With Collagenous Gastritis.
Collagenous gastritis is a rare form of gastritis that affects both children and adults. The underlying pathophysiology is not well-understood, and as a result, there are limited options for treatment. We report a case of a young female patient with chronic diffuse abdominal pain, nausea, regurgitation, and early satiety with esophagogastroduodenoscopy showing gastric erythema, atrophic gastric body, and significant gastric nodularity. Biopsies revealed focal erosion and increased subepithelial collagen deposition. She was successfully managed with intravenous vedolizumab infusions after initial therapy with topical budesonide did not result in clinical or endoscopic improvement.


Pheomelanin coat colour dilution in French cattle breeds is not correlated with the TYR, TYRP1 and DCT transcription levels.
In this study we report the isolation of full-length cDNAs and the expression patterns of TYR, TYRP1 and DCT in four e/e cattle breeds exhibiting different pheomelanic coat colours ranging from reddish brown to creamy white phenotypes. Predicted proteins encoded by bovine TYR, TYRP1 and DCT display high levels of homology and contain all characteristic domains shared between their mouse and human counterparts. The full expression of these three genes is observed in melanocytes of black areas of E(D)/E(D) Prim'Holstein's animals. On the other hand, e/e melanocytes of animals belonging to the Blonde d'Aquitaine (blond), Limousine (red) and Salers (reddish brown) breeds present different levels of down-regulated TYR and DCT expression and a complete repression of TYRP1. Surprisingly, e/e melanocytes of animals belonging to the Charolais breed (creamy white) present an inverse relationship between TYR, TYRP1 and DCT expression and its lower melanogenic activity. The sum of these results shows that the dilution of the coat colour in French cattle breeds is not correlated with a transcription level of TYR family genes. Other possible modifier loci are suggested.


Measuring the temperature of microparticles in plasmas.
Temperature sensitive features of particular phosphors were utilized for measuring the temperature T(p) of microparticles, confined in the sheath of a rf plasma. The experiments were performed under variation of argon pressure and rf power of the process plasma. T(p) has been determined by evaluation of characteristic fluorescent lines. The results for T(p) measurements are strongly dependent on rf power and gas pressure.


[Effects of cardiac support on delayed resuscitation in extensively burned patients with shock].
Objective: To explore the effects of cardiac support on delayed resuscitation in extensively burned patients with shock. Methods: Clinical data of 62 extensively burned patients with shock on admission, admitted to the 159th Hospital of PLA (hereinafter referred to as our hospital) from January 2012 to January 2017, were retrospectively analyzed. They were divided into cardiac support group (n=35) and control group (n=27) according to the use of deslanoside and ulinastatin. All patients were treated with routine fluid resuscitation based on the formula of the Third Military Medical University till post injury hour (PIH) 48. Patients in cardiac support group were given slow intravenous injection of deslanoside which was added in 20 mL 100 g/L glucose injection with first dose of 0.4 to 0.6 mg, 0.2 to 0.4 mg per 6 to 8 h, no more than 1.6 mg daily, and slow intravenous injection of 1×10(5)U ulinastatin which was added in 100 mL 50 g/L glucose injection, once per 12 h. Other treatments of patients in the two groups followed the same conventional procedures of our hospital. The following data of the two groups of patients were collected. (1) The data of urine volume per hour within PIH 48, heart rate, mean arterial pressure (MAP), central venous pressure (CVP), blood lactic acid, base excess, hematocrit, and albumin at PIH 48 were recorded. (2) The input volumes of electrolyte, colloid within the first and second 24 hours post burn and the total fluid input volumes within PIH 48 were recorded. (3) The data of creatine kinase, creatine kinase isoenzyme-MB, lactate dehydrogenase, total bile acid, alanine aminotransferase, aspartate aminotransferase, β(2)-microglobulin, urea nitrogen, and creatinine at PIH 48 were recorded. (4) The complications including cardiac failure, pulmonary edema, pleural effusion, seroperitoneum, renal failure, sepsis, and death were also recorded. Data were processed with independent sample ttest, Fisher's exact test, Pearson chi-square test, or continuous correction chi-square test. Results: (1) There were no statistically significant differences in urine volume within PIH 48, heart rate, MAP, CVP, hematocrit, or albumin at PIH 48 between the patients of two groups (t=0.150, 0.488, 0.805, 0.562, 1.742, 0.696, P>0.05). While the levels of blood lactic acid and base excess were respectively (4.2±2.2) and (-4.3±2.0) mmol/L in patients of cardiac support group, which were significantly better than (5.9±1.7) and (-6.0±3.1) mmol/L in patients of control group (t=3.249, 2.480, P<0.05 or P<0.01). (2) There was no statistically significant difference in input volume of colloid within the first 24 hours post burn between the patients of two groups (t=0.642, P>0.05). The input volume of electrolyte within the first 24 hours post burn, the input volumes of electrolyte and colloid within the second 24 hours post burn, and the total fluid input volume within PIH 48 of patients in cardiac support group were significantly less than those in control group (t=2.703, 4.223, 3.437, 2.515, P<0.05 or P<0.01). (3) The levels of creatine kinase, creatine kinase isoenzyme-MB, lactate dehydrogenase, total bile acid, alanine aminotransferase, aspartate aminotransferase, β(2)-microglobulin, urea nitrogen, and creatinine of patients in cardiac support group at PIH 48 were significantly lower than those in control group (t=3.066, 3.963, 3.225, 2.943, 2.431, 3.084, 4.052, 2.915, 3.353, P<0.05 or P<0.01). (4) The occurrences of pleural effusion and seroperitoneum and mortality of patients in cardiac support group were significantly lower than those in control group (χ(2)=5.514, 6.984, 4.798, P<0.05 or P<0.01). There were no statistically significant differences in cardiac failure, pulmonary edema, renal failure, and sepsis between the patients of two groups [χ(2)=1.314 (sepsis), P>0.05]. Conclusions: The cardiotonic and cardiac protection treatments in delayed resuscitation of extensively burned patients with shock contribute to improving the cellular anonic metabolism, reducing the volume of fluid resuscitation, and mitigating the ischemic and hypoxic damage to organs, so as to lay foundation for decreasing further complication incidences and mortality.


Prescribing cost savings by GP fundholders: long-term or short-term?
Fundholding general practices have been observed to be more successful than non-fundholders in controlling the growth of their prescribing costs. Debate persists over the likely duration of this fundholding effect. Regression analysis of changes in prescribing costs for a large sample of practices over 5 years supports the view that prescribing cost economies have been short-rather than long-term, and that practices entering fundholding did not engage in strategic, cost-raising behaviour prior to joining the scheme.


Do PICU patients meet technical criteria for performing indirect calorimetry?
Indirect calorimetry (IC) is considered gold standard for assessing energy needs of critically ill children as predictive equations and clinical status indicators are often unreliable. Accurate assessment of energy requirements in this vulnerable population is essential given the high risk of over or underfeeding and the consequences thereof. The proportion of patients and patient days in pediatric intensive care (PICU) for which energy expenditure (EE) can be measured using IC is currently unknown. In the current study, we aimed to quantify the daily proportion of consecutive PICU patients who met technical criteria to perform indirect calorimetry and describe the technical contraindications when criteria were not met. Prospective, observational, single-centre study conducted in a cardiac and general PICU. All consecutive patients admitted for at least 96 h were included in the study. Variables collected for each patient included age at admission, admission diagnosis, and if technical criteria for indirect calorimetry were met. Technical criteria variables were collected within the same 2 h each morning and include: provision of supplemental oxygen, ventilator settings, endotracheal tube (ETT) leak, diagnosis of chest tube air leak, provision of external gas support (i.e. nitric oxide), and provision of extracorporeal membrane oxygenation (ECMO). 288 patients were included for a total of 3590 patient days between June 2014 and February 2015. The main reasons for admission were: surgery (cardiac and non-cardiac), respiratory distress, trauma, oncology and medicine/other. The median (interquartile range) patient age was 0.7 (0.3-4.6) years. The median length of PICU stay was 7 (5-14) days. Only 34% (95% CI, 32.4-35.5%) of patient days met technical criteria for IC. For patients less than 6 months of age, technical criteria were met on significantly fewer patient days (29%, p < 0.01). Moreover, 27% of patients did not meet technical criteria for IC on any day during their PICU stay. Most frequent reasons for why IC could not be performed included supplemental oxygen, ECMO, and ETT leak. In the current study, technical criteria to perform IC in the PICU were not met for 27% of patients and were not met on 66% of patient days. Moreover, criteria were met on only 29% of days for infants 6 months and younger where children 24 months of age and older still only met criteria on 40% of patient days. This data represents a major gap in the feasibility of current recommendations for assessing energy requirements of this population. Future studies are needed to improve methods of predicting and measuring energy requirements in critically ill children who do not meet current criteria for indirect calorimetry.


Modelling Down syndrome.
Animal models are extensively used in genetics, neuroscience and biomedical research. Recent studies illustrate the usefulness and the challenges of research utilising genetically engineered mice to explore the developmental biology of Down syndrome. These studies highlight many of the issues at the centre of what we understand about Down syndrome, and may one day point to useful ways to improve quality of life for people living with Down syndrome.


Beaming and enhanced transmission through a subwavelength aperture via epsilon-near-zero media.
We numerically validate and experimentally realize considerable funneling of electromagnetic energy through a subwavelength aperture that is covered with an epsilon-near-zero metamaterial (ENZ). The epsilon-near-zero metamaterial is composed of two layers of metasurfaces and operates at microwave frequencies. We demonstrate that the presence of the metamaterial at the inner and outer sides of the aperture not only lead to a significant enhancement in light transmission, but also cause a directional emission of light extracting from this hybrid system. In addition to these experimental results, we theoretically demonstrate the same concept in mid-IR region for a subwavelength gold aperture with indium tin oxide as an epsilon-near-zero material. Moreover, we found that using a dielectric spacer in-between the sunwavelength aperture and the ENZ medium, it is possible to red-shift the enhancement/directional frequency of the system.


RNA-binding proteins that are highly expressed and enriched in healthy cartilage but suppressed in osteoarthritis.
Objectives: RNA-binding proteins (RBPs) have diverse and essential biological functions, but their role in cartilage health and disease is largely unknown. The objectives of this study were (i) map the global landscape of RBPs expressed and enriched in healthy cartilage and dysregulated in osteoarthritis (OA); (ii) prioritize RBPs for their potential role in cartilage and in OA pathogenesis and as therapeutic targets. Methods: Our published bulk RNA-sequencing (RNA-seq) data of healthy and OA human cartilage, and a census of 1,542 RBPs were utilized to identify RBPs that are expressed in healthy cartilage and differentially expressed (DE) in OA. Next, our comparison of healthy cartilage RNA-seq data to 37 transcriptomes in the Genotype-Tissue Expression (GTEx) database was used to determine RBPs that are enriched in cartilage. Finally, expression of RBPs was analyzed in our single cell RNA-sequencing (scRNA-seq) data from healthy and OA human cartilage. Results: Expression of RBPs was higher than nonRBPs in healthy cartilage. In OA cartilage, 188 RBPs were differentially expressed, with a greater proportion downregulated. Ribosome biogenesis was enriched in the upregulated RBPs, while splicing and transport were enriched in the downregulated. To further prioritize RBPs, we selected the top 10% expressed RBPs in healthy cartilage and those that were cartilage-enriched according to GTEx. Intersecting these criteria, we identified Tetrachlorodibenzodioxin (TCDD) Inducible Poly (ADP-Ribose) Polymerase (TIPARP) as a candidate RBP. TIPARP was downregulated in OA. scRNA-seq data revealed TIPARP was most significantly downregulated in the "pathogenic cluster". Conclusion: Our global analyses reveal expression patterns of RBPs in healthy and OA cartilage. We also identified TIPARP and other RBPs as novel mediators in OA pathogenesis and as potential therapeutic targets.


Terminal Deoxynucleotidyl Transferase-Mediated Deoxyuridine Triphosphate Nick End Labeling (TUNEL) Assay to Characterize Histopathologic Changes Following Thermal Injury.
Despite the wide application of lasers and radiofrequency (RF) surgery in dermatology, it is difficult to find studies showing the extent of damage dependent on cell death. We evaluated histopathologic changes following in vivo thermal damage generated by CO2 laser, 1,444 nm long-pulsed neodymium:yttrium-aluminum-garnet (LP Nd:YAG) laser and RF emitting electrosurgical unit. Thermal damage was induced by the above instruments on ventral skin of rat. Specimens were stained with hematoxylin and eosin, along with a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay, to highlight the degree of irreversible cellular injury. The volume of vaporization was largest with the CO2 laser. Area of cell death area identified by TUNEL assay, when arranged from widest to narrowest, was 1,444 nm LP Nd:YAG laser, CO2 laser, and RF emitting electrosurgical unit. This histopathologic evaluation of the acute characterization of injury across devices may be advantageous for attaining better treatment outcomes.


Adjunctive techniques for oral cancer examination and lesion diagnosis: a systematic review of the literature.
Adjunctive techniques that may facilitate the early detection of oral premalignant and malignant lesions (OPML) have emerged in the past decades. The authors undertook a systematic review of the English-language literature to evaluate the effectiveness of toluidine blue (TB), ViziLite Plus with TBlue (Zila Pharmaceuticals, Phoenix), ViziLite (Zila Pharmaceuticals), Microlux DL (AdDent, Danbury, Conn.), Orascoptic DK (Orascoptic, a Kerr Company, Middleton, Wis.), VELscope (LED Dental, White Rock, British Columbia, Canada) and OralCDx (Oral CDx Laboratories, Suffern, N.Y.) brush biopsy. They abstracted data relating to study design, sampling and characteristics of the study group, interventions, reported outcomes and diagnostic accuracy of adjunctive aids from 23 articles meeting inclusion and exclusion criteria, including availability of histologic outcomes. The largest evidence base was for TB. A limited number of studies was available for ViziLite, ViziLite Plus with TBlue and OralCDx. Studies of VELscope have been conducted primarily to assess the margins of lesions in known OPML. The authors identified no studies of Microlux DL or Orascoptic DK. Study designs had various limitations in applicability to the general practice setting, including use of higher-risk populations and expert examiners. There is evidence that TB is effective as a diagnostic adjunct for use in high-risk populations and suspicious mucosal lesions. OralCDx is useful in assessment of dysplastic changes in clinically suspicious lesions; however, there are insufficient data meeting the inclusion criteria to assess usefulness in innocuous mucosal lesions. Overall, there is insufficient evidence to support or refute the use of visually based examination adjuncts. Practical Implications. Given the lack of data on the effectiveness of adjunctive cancer detection techniques in general dental practice settings, clinicians must rely on a thorough oral mucosal examination supported by specialty referral and/or tissue biopsy for OPML diagnosis.


Endothelium-dependent relaxation in isolated pulmonary arteries from rabbits exposed to hyperoxia.
The effects of hyperoxia and superoxide dismutase (SOD) administration on endothelium-dependent relaxation induced by acetylcholine in isolated rabbit pulmonary artery (PA) preparations were examined. Relaxation responses to papaverine, and contractile responses to KCl and norepinephrine were also assessed using a bioassay technique. Prolonged hyperoxia in rabbits for 3 days produced loss of endothelium-dependent relaxation of the PA, and significant attentuation of contractile response to KCl and relaxation response to papaverine. These changes were prevented by subcutaneous SOD administration. Histological examination using light and electron microscopy revealed focal edema, destruction, and detachment of the PA endothelium in the PA strip preparations from these rabbits. Thus, it is concluded that a high concentration of oxygen exposure in rabbits for 3 days produces not only histological damage in the PA endothelium, but also causes impairment of vascular reactivity to constricting and relaxing agents. Subcutaneous SOD administration prevented oxygen-induced PA damage.


Colorectal cancer in younger population: our experience.
To promote awareness regarding increased occurrence of colorectal cancer in younger population and its clinicopathological features compared to older patients. The cross-sectional study was conducted from February 2010 to January 2011 on patients with diagnosis of colorectal carcinoma admitted through emergency or outpatients' departments to Surgical Unit 5, Civil Hospital, Karachi. Data regarding age, gender, presentation, site of tumour, surgery performed and Dukes staging was collected and analysed. A total of 23 patients were operated during the study period: 13 (56.52%) males and 10 (43.47%) females. Of them 12 (52.17%) were below the age of 40 years, while 3 (13.04%) patients were in the 11-20 age group. In 7 (30.4%) patients, tumour was irresectable at the time of presentation so a palliative procedure (diversion colostomy or ileostomy) was performed.There was a higher proportion of younger patients with metastatic disease at the time of presentation (n = 9; 75%) while 10 out of 12 patients in the younger age group (83.3%) had a tumour of left colon, particularly rectum. Although colorectal cancer is usually a disease of older patients, it is increasingly becoming more common in younger population. Data suggests a leftward distribution for colorectal carcinoma and that younger patients present with more advanced disease and poorer prognosis.


The genetic basis for variation in resistance to infection in the Drosophila melanogaster genetic reference panel.
Individuals vary extensively in the way they respond to disease but the genetic basis of this variation is not fully understood. We found substantial individual variation in resistance and tolerance to the fungal pathogen Metarhizium anisopliae Ma549 using the Drosophila melanogaster Genetic Reference Panel (DGRP). In addition, we found that host defense to Ma549 was correlated with defense to the bacterium Pseudomonas aeruginosa Pa14, and several previously published DGRP phenotypes including oxidative stress sensitivity, starvation stress resistance, hemolymph glucose levels, and sleep indices. We identified polymorphisms associated with differences between lines in both their mean survival times and microenvironmental plasticity, suggesting that lines differ in their ability to adapt to variable pathogen exposures. The majority of polymorphisms increasing resistance to Ma549 were sex biased, located in non-coding regions, had moderately large effect and were rare, suggesting that there is a general cost to defense. Nevertheless, host defense was not negatively correlated with overall longevity and fecundity. In contrast to Ma549, minor alleles were concentrated in the most Pa14-susceptible as well as the most Pa14-resistant lines. A pathway based analysis revealed a network of Pa14 and Ma549-resistance genes that are functionally connected through processes that encompass phagocytosis and engulfment, cell mobility, intermediary metabolism, protein phosphorylation, axon guidance, response to DNA damage, and drug metabolism. Functional testing with insertional mutagenesis lines indicates that 12/13 candidate genes tested influence susceptibility to Ma549. Many candidate genes have homologs identified in studies of human disease, suggesting that genes affecting variation in susceptibility are conserved across species.


Data-dependent middle-down nano-liquid chromatography-electron capture dissociation-tandem mass spectrometry: an application for the analysis of unfractionated histones.
Middle-down mass spectrometry (MS) combined with electron capture dissociation (ECD) represents an attractive method for characterization of proteins and their post-translational modifications (PTMs). Coupling online chromatographic separation with tandem mass spectrometry enables a high-throughput analysis, while improving sensitivity of the electrosprayed peptides and reducing sample amount requirements. However, middle-down ECD has not been thus far coupled with online chromatographic separation. In this work, we examine the feasibility of coupling middle-down ECD with online nanoflow-liqiud chromatography (nano-LC) for the analysis of large, >3 kDa, and highly modified polypeptides in a data-dependent acquisition mode. We evaluate the effectiveness of the method by analyzing peptides derived from Asp-N and Glu-C digestions of unfractionated histones from calf thymus and acid-extracted histones from HeLa, MCF-7, and Jurkat cells. Our results demonstrate that middle-down ECD is compatible with online chromatographic separation, providing high peptide and protein sequence coverage while allowing precise mapping of PTM sites. The high mass accuracy, obtained by the ICR mass analyzer, for both the precursor and product ions greatly increases confidence in peptide identification, particularly for modified peptides. Overall, for all samples examined, several histone variants were identified and modification sites were successfully localized, including single, multiple, and positional isomeric PTM sites. The vast majority of the identified peptides were in the mass range from 3 to 9 kDa. The data presented here highlight the feasibility and utility of nano-LC-ECD-MS/MS for high-throughput middle-down analysis.


Inpatient health care utilization in the United States among children, adolescents, and young adults with nephrotic syndrome.
Data describing inpatient health care utilization in children with nephrotic syndrome and related severe complications are limited. Our goals were to describe the charges, length of stay (LOS), and number of hospitalizations among children, adolescents, and young adults with nephrotic syndrome. A cross-sectional analysis of the Kids' Inpatient Database (KID) database from the Healthcare Cost and Utilization Project (HCUP). The HCUP-KID is an all-payer database of hospital discharges for children, adolescents, and young adults in the United States compiled every 3 years by the Agency for Healthcare Research and Quality. HCUP-KID data were obtained for the 2006 and 2009 cohort years. We identified patients by searching discharges for nephrotic syndrome International Classification of Diseases, Ninth Revision, Clinical Modification diagnosis codes. Patient demographics, disease complications in children, adolescents, and young adults hospitalized with nephrotic syndrome. Number of hospitalizations, mean charges, and LOS for children, adolescents, and young adults hospitalized with nephrotic syndrome. There were 6,308 hospitalization discharges in children, adolescents, and young adults with a primary or secondary diagnosis of nephrotic syndrome reported by 38 and 44 states in 2006 and 2009, respectively, representing an estimated 9,934 discharges nationally. Nephrotic syndrome resulted in an estimated 48,700 inpatient days and charges totaling $259 million. The mean charge per hospitalization was ∼$26,500 (SE, $1,100) and LOS was 5 days (SE, 0.1). 16% of discharges for nephrotic syndrome had a diagnosis code for at least one severe complication, including thromboembolism (3.6%), septicemia (3.8%), peritonitis (2.6%), pneumonia (5.4%), or diabetes (2.4%). Multivariable analysis showed age 15 years or older, race, higher socioeconomic status, acute renal failure, thromboembolic disease, hypertension, and infections predicted higher mean hospitalization charges. The HCUP-KID database collects data on a hospitalization level. Consequently, health care utilization on an individual patient level or in the outpatient environment is not possible. We present a comprehensive description of inpatient health care utilization in children, adolescents, and young adults with nephrotic syndrome. The complications of nephrotic syndrome, including thromboembolism, infection, and hypertension, contribute significantly to these charges.


Origin and nature of bond rotation barriers: a unified view.
Bond rotations are common and important in molecular studies such as drug design and enzymatic reactions. Steric hindrance and hyperconjugation are often borrowed to justify the origin of bond rotation barriers. However, there exists no consensus, even for as simple molecules as ethane, on which the effect is more significant. Here, we show that a unified view is possible. To that end, we employ two energy partition schemes from density functional theory to investigate the flexible rotation barrier of six molecules with one rotatable dihedral angle: ethane, methylamine, methanol, hydrazine, hydroxylamine, and hydrogen peroxide. Our results suggest that, even though steric and quantum effects play indispensable roles, it is the electrostatic interaction that governs the barrier height of all these different types of systems. This work not only consolidates earlier views about the role of steric and quantum effects, it also provides new insights about the origin and nature of the bond rotation barrier, which should be applicable to many different types of chemical bonds.


Characteristics and Predictors of Readmission to a Medical Short-Stay Unit.
Short-stay units are medical units with anticipated lengths of stay of 24-72 hours. The purpose of this study was to identify characteristics and predictors of 30-day readmission among patients admitted to a short-stay unit during two separate time periods. Findings suggest the presence of previous Emergency Department visits or hospitalizations should be considered when aligning discharge needs, as these may impact subsequent patient readmissions.


Exploring district nurses' experience of a hospice at home service.
this study surveyed a representative sample of district nurses (DNs) to ascertain their experience of working alongside a HAH service. a self-completed postal questionnaire comprised of 14 items was distributed to 128 DNs. the evaluation found significant levels of satisfaction. All respondents indicated that they would refer a patient to the service again. However, the need for improved communication and increased awareness among DNs about the HAH service were also identified. the findings support the idea that a HAH service can enable patients with advanced progressive disease to be cared for at home. While the findings of this study cannot be generalized, they add to the growing body of research about HAH services in palliative care.


Specific COVID-19 Symptoms Correlate with High Antibody Levels against SARS-CoV-2.
Lasting immunity will be critical for overcoming COVID-19. However, the factors associated with the development of high titers of anti-SARS-CoV-2 Abs and how long those Abs persist remain incompletely defined. In particular, an understanding of the relationship between COVID-19 symptoms and anti-SARS-CoV-2 Abs is limited. To address these unknowns, we quantified serum anti-SARS- CoV-2 Abs in clinically diverse COVID-19 convalescent human subjects 5 wk (n = 113) and 3 mo (n = 79) after symptom resolution with three methods: a novel multiplex assay to quantify IgG against four SARS-CoV-2 Ags, a new SARS-CoV-2 receptor binding domain-angiotensin converting enzyme 2 inhibition assay, and a SARS-CoV-2 neutralizing assay. We then identified clinical and demographic factors, including never-before-assessed COVID-19 symptoms, that consistently correlate with high anti-SARS-CoV-2 Ab levels. We detected anti-SARS-CoV-2 Abs in 98% of COVID-19 convalescent subjects 5 wk after symptom resolution, and Ab levels did not decline at 3 mo. Greater disease severity, older age, male sex, higher body mass index, and higher Charlson Comorbidity Index score correlated with increased anti-SARS-CoV-2 Ab levels. Moreover, we report for the first time (to our knowledge) that COVID-19 symptoms, most consistently fever, body aches, and low appetite, correlate with higher anti-SARS-CoV-2 Ab levels. Our results provide robust and new insights into the development and persistence of anti-SARS-CoV-2 Abs.