[Use of nasal mupirocin for Staphylococcus aureus: effect on nasal carriers and nosocomial infections].
Staphylococcus aureus is the agent of community-acquired and nosocomial infections. Twenty to 35% of the population permanently carries it in the nose and oropharynx, and additional 50%, carries it intermittently. Topical calcium mupirocin is an antibacterial agent against Staphylococcus aureus recommended to eradicate nasal and hand colonization in patients and health care workers. The prevalence of nasal S. aureus was determined in patients undergoing cardiovascular surgery. In addition, the effect of mupirocine on the number of carriers and rate of nosocomial infections was evaluated. An experimental prospective study was undertaken with two groups of patients: one treated with mupirocin (n = 96), and the other without treatment (n = 95). Tests for presence of nasal S. aureus and nosocomial infections were conducted in all patients. A 34% prevalence of S. aureus carriers was found. A decrease of the prevalence was found in both treated (87%) and untreated patients (33%), but in significantly different proportions (p = 0.0002, RR = 0.22, 95%CI = 0.09-0.054). This result demonstrated the effectiveness of a mupirocin treatment program to decrease numbers of nasal carriers. With regard to nosocomial infection, S. aureus prevalence was 3.6%, occurring mostly in control patients (6 of 7). Total nosocomial infection prevalence was 17.3%, evenly distributed in treated and untreated patients. This suggested that mupirocin use did not decrease the number of nosocomial infections.


Chaos-on-a-chip secures data transmission in optical fiber links.
Security in information exchange plays a central role in the deployment of modern communication systems. Besides algorithms, chaos is exploited as a real-time high-speed data encryption technique which enhances the security at the hardware level of optical networks. In this work, compact, fully controllable and stably operating monolithic photonic integrated circuits (PICs) that generate broadband chaotic optical signals are incorporated in chaos-encoded optical transmission systems. Data sequences with rates up to 2.5 Gb/s with small amplitudes are completely encrypted within these chaotic carriers. Only authorized counterparts, supplied with identical chaos generating PICs that are able to synchronize and reproduce the same carriers, can benefit from data exchange with bit-rates up to 2.5Gb/s with error rates below 10(-12). Eavesdroppers with access to the communication link experience a 0.5 probability to detect correctly each bit by direct signal detection, while eavesdroppers supplied with even slightly unmatched hardware receivers are restricted to data extraction error rates well above 10(-3).


Learning a saliency map using fixated locations in natural scenes.
Inspired by the primate visual system, computational saliency models decompose visual input into a set of feature maps across spatial scales in a number of pre-specified channels. The outputs of these feature maps are summed to yield the final saliency map. Here we use a least square technique to learn the weights associated with these maps from subjects freely fixating natural scenes drawn from four recent eye-tracking data sets. Depending on the data set, the weights can be quite different, with the face and orientation channels usually more important than color and intensity channels. Inter-subject differences are negligible. We also model a bias toward fixating at the center of images and consider both time-varying and constant factors that contribute to this bias. To compensate for the inadequacy of the standard method to judge performance (area under the ROC curve), we use two other metrics to comprehensively assess performance. Although our model retains the basic structure of the standard saliency model, it outperforms several state-of-the-art saliency algorithms. Furthermore, the simple structure makes the results applicable to numerous studies in psychophysics and physiology and leads to an extremely easy implementation for real-world applications.


Periods of organogenesis in shoots of Nothofagus dombeyi (Mirb.) oersted (Nothofagaceae).
The organogenetic cycle of main-branch shoots of Nothofagus dombeyi (Nothofagaceae) was studied. Twelve samples of 52-59 parent shoots were collected from a roadside population between September 1999 and October 2000. Variations over time in the number of nodes of terminal and axillary buds, and the length, diameter and number of leaves of shoots derived from these buds (sibling shoots) were analysed. The number of nodes of buds developed by parent shoots was compared with the number of nodes of buds developed, I year later, by sibling shoots. The length, diameter and number of leaves of sibling shoots increased from October 1999 to February 2000 in those shoots with a terminal bud. However, extension of most sibling shoots, including the first five most distal leaf primordia, ceased before February due to abscission of the shoot apex. Axillary buds located most distally on a shoot had more nodes than both terminal buds and more proximal axillary buds. The longest shoots included a preformed part and a neoformed part. The organogenetic event which initiated the neoformed organs continued until early autumn, giving rise to the following year's preformation. The absence of cataphylls in terminal buds could indicate a low intensity of shoot rest. The naked terminal bud of Nothofagus spp. could be interpreted as a structure less specialized than the scaled bud found in genera of Fagaceae and Betulaceae.


Relationship between CYP2C9 and 2C19 genotypes and tolbutamide methyl hydroxylation and S-mephenytoin 4'-hydroxylation activities in livers of Japanese and Caucasian populations.
Genomic DNA was isolated from livers of 39 Japanese and 45 Caucasians and the genotypes of CYP2C9 and 2C19 genes were determined with PCR methods using synthetic oligonucleotide primers. Liver microsomes were also prepared from these human samples and activities for tolbutamide methyl hydroxylation and S-mephenytoin 4'-hydroxylation were determined. The single base mutation of C416T (Arg144Cys) in CYP2C9 was detected in 22% of Caucasians but not in Japanese samples. Another single base mutation at A1061C (Ile359Leu) in the CYP2C9 gene was found with frequencies of about 8% in both races. We did not detect any individuals who have either homozygous Cys144/Cys144 or Leu359/Leu359 CYP2C9 variant nor both heterozygous Cys144-Ile359 and Arg144-Leu359 CYP2C9 variant in the human samples examined. The CYP2C19m2 genetic polymorphism was found only in Japanese people, while CYP2C19m1 type was determined in both races, with higher incidence in Japanese than in Caucasian population. Immunoblotting analysis of human liver microsomes suggested that CYP2C9 is a major component of the human CYP2C enzyme pool; it accounted for approximately 20% of total P450 in liver microsomes of both human populations. The levels of CYP2C19 protein were determined to be about 0.8% and 1.4% of total P450 (mean) in Japanese and Caucasians, respectively. We did not detect CYP2C19 protein in liver microsomes of humans who were genotyped for CYP2C19 gene as m1/m1, m1/m2, and m2/m2 variants but detected CYP2C9 protein in all of the samples examined. Good correlations were found between levels of CYP2C9 and activities of tolbutamide methyl hydroxylation (r = 0.77) and between levels of CYP2C19 and activities of S-mephenytoin 4'-hydroxylation (r = 0.86) in liver microsomes of the human samples examined. Tolbutamide methyl hydroxylation activities were lower in human samples with the Leu359 allele of CYP2C9 than those with the Cys144 allele and wild-type (Arg144-Ile359); the former type showed slightly higher K(m) values. When calculated on P450 basis, liver microsomes of individuals having m1/m1, m1/m2, and m2/m2 types of CYP2C19 had very low catalytic activities for S-mephenytoin 4'-hydroxylation. These results provide useful comparisons for pharmacokinetic and toxicokinetic models of some of the clinically used drugs that are oxidized by CYP2C proteins in humans.


Finger probe array for topography-tolerant scanning electrochemical microscopy of extended samples.
Scanning electrochemical microscopy with soft microelectrode array probes has recently been used to enable reactivity imaging of extended areas and to compensate sample corrugation perpendicular to the scanning direction. Here, the use of a new type of microelectrode arrays is described in which each individual microelectrode can independently compensate corrugations of the sample surface. It consists of conventional Pt microelectrodes enclosed in an insulating glass sheath. The microelectrodes are individually fixed to a new holder system by magnetic forces. The concept was tested using a large 3D sample with heights up to 12 μm specially prepared by inkjet printing. The microelectrodes follow the topography in a constant working distance independently from each other while exerting low pressure on the surface.


[Intrafollicular infection of mammals and human oocytes by the herpes simplex virus].
The goal of this work was to study the capacity of the herpes simplex virus (HSV) of infecting ovary with disease in case of the intravaginal experimental animals. The results of the study demonstrated that the ascending HSV infection in mice lead to modification of all the cells of the ovary, including follicular cells synthesizing estrogen and progesterone. The two hormones influence the development of the disease. Estrogens provide the protective effects against the virus. Progesterone does not modify the body sensitivity to HSV, but reduces the effectiveness of the antiviral immunity, resulting in increased mortality of animals. We demonstrated that infection of oocytes in ovarian follicles of female mice during infection with HSV modified the process in vitro and for the first time demonstrated the detection of viral antigens in mature oocytes in patient with infertility. During the intracytoplasmic sperm injection into the infected oocytes (ICSI), the failure of fertilization was observed. These results are of interest, because there is no available literature on whether HSV infection of oocytes can have a direct negative impact on the process of fertilization in humans.


Salivary, nasal, genital, and systemic antibody responses in monkeys immunized intranasally with a bacterial protein antigen and the Cholera toxin B subunit.
Previous attempts to induce mucosal antibodies in rhesus monkeys by enteric immunization have resulted in only modest and short-lived responses, dominated by immunoglobulin M (IgM) antibodies in the plasma. In this study, two groups of rhesus monkeys were immunized intranasally three times at 2-week intervals with a bacterial protein antigen (AgI/II) either chemically coupled to or mixed with the B subunit of cholera toxin (CT), a known potent mucosal immunogen and carrier for other immunogens. Cells secreting antibodies, predominantly of the IgA isotype, to AgI/II and to CT were detected in the peripheral blood 1 week after each immunization, indicating the dissemination of IgA-secreting precursor cells through the mucosal immune system. IgG and, to a lesser extent, IgA antibodies to both proteins were induced in the plasma commencing after the second immunization. Plasma IgE concentrations and IgE antibody levels were not consistently raised during the immunization period. IgA antibodies were found in nasal and vaginal washes. Nasal IgG but not IgA antibodies showed a significant positive correlation with plasma IgG antibody levels, suggesting that they were largely derived by transudation from the circulation. Analysis of the molecular form of vaginal IgA indicated that both secretory and monomeric forms of IgA were present in various proportions. Furthermore, neither IgG nor IgA antibodies in vaginal washes were correlated with plasma antibody responses, suggesting the contribution of locally synthesized antibodies of both isotypes. Comparison of the responses between the two groups of animals showed only sporadic significant differences, indicating that intranasal immunization with AgI/II either coupled to or mixed with the B subunit of CT was equally effective at inducing generalized IgA antibody responses in the mucosal immune system and predominantly IgG antibodies in the plasma.


13 C metabolic flux profiling of Pichia pastoris grown in aerobic batch cultures on glucose revealed high relative anabolic use of TCA cycle and limited incorporation of provided precursors of branched-chain amino acids.
Carbon metabolism of Crabtree-negative yeast Pichia pastoris was profiled using 13 C nuclear magnetic resonance (NMR) to delineate regulation during exponential growth and to study the import of two precursors for branched-chain amino acid biosynthesis, α-ketoisovalerate and α-ketobutyrate. Cells were grown in aerobic batch cultures containing (a) only glucose, (b) glucose along with the precursors, or (c) glucose and Val. The study provided the following new insights. First, 13 C flux ratio analyses of central metabolism reveal an unexpectedly high anaplerotic supply of the tricarboxylic acid cycle for a Crabtree-negative yeast, and show that a substantial fraction of glucose catabolism proceeds through the pentose phosphate pathway. A comparison with previous flux ratio analyses for batch cultures of Crabtree-negative Pichia stipitis and Crabtree-positive Saccharomyces cerevisiae indicate that the overall regulation of central carbon metabolism in P. pastoris is intermediate in between P. stipitis and S. cerevisiae. Second, excess α-ketoisovalerate in the medium is not transported into the cytoplasm indicating that P. pastoris lacks a suitable transporter. In contrast, excess Val is efficiently taken up and largely fulfills demands for both Val and Leu for protein synthesis. Third, excess α-ketobutyrate is transported into the mitochondria for Ile biosynthesis. However, the import does not efficiently inhibit the synthesis of α-ketobutyrate from pyruvate indicating that P. pastoris has not been optimized evolutionarily to take full advantage of this carbon source. These findings have direct implications for preparing uniformly 2 H,13 C,15 N-labeled proteins containing protonated Ile, Val, and Leu methyl groups in P. pastoris for NMR-based structural biology. Acetohydroxy acid isomeroreductase (EC 188.8.131.52), branched-chain amino acid aminotransferase (BCAT, EC 184.108.40.206), fumarase (EC 188.8.131.52), malic enzyme (EC 126.96.36.199/184.108.40.206), phosphoenolpyruvate carboxykinase (EC 126.96.36.199), pyruvate carboxylase (EC 220.127.116.11), pyruvate kinase (EC 126.96.36.199), l-serine hydroxymethyltransferase (EC 220.127.116.11), threonine aldolase (EC 184.108.40.206), threonine dehydratase (EC 188.8.131.52); transketolase (EC 188.8.131.52), transaldolase (EC 18.104.22.168).


Effect of immunization against rubella on lactation products. I. Development and characterization of specific immunologic reactivity in breast milk.
The development of an immune response to rubella virus in milk, serum, and nasopharyngeal secretions was studied in lactating postpartum women after immunization with HPV-77 De5 or RA 27/3 live, attenuated rubella virus vaccine administered subcutaneously or intranasally. Over 69% of the women shed virus in milk after immunization. A predictable nasopharyngeal IgA and serum IgG antibody response to rubella virus was observed after subcutaneous or intranasal immunization with RA 27/3 vaccine. Little or no nasopharyngeal antibody response was seen after subcutaneous immunization with HPV-77 DE5 vaccine. A virus-specific IgA antibody response in milk was seen in all women. The presence of rubella virus in breast milk seemed to potentiate the peak levels of virus-specific antibody in the milk. Cellular immune reactivity to rubella virus in milk was observed in all vaccine groups. Thus, the virus-specific immune response induced in human milk after immunization with rubella virus vaccine may be intimately linked with the reactivity in bronchial lymphoid tissue.


Assessing Alignment Using GAP Score and Complications for Pedicle Subtraction Osteotomy Revision Surgeries for Sagittal Deformity in Previously Fused Spines Using a Satellite Rod Technique.
Pedicle subtraction osteotomy (PSO) is a surgical technique to restore lumbar lordosis in patients with rigid sagittal deformity. Herein, we report on outcomes of a surgical technique utilizing a 4-rod PSO with satellite rods. A retrospective study was performed for a cohort of patients who underwent 4-rod PSO revision surgery for sagittal deformity. Procedures were performed by one surgeon at 2 different academic centers between 2004-2018. Alignment outcomes and complications specifically for revision surgeries in previously fused spines were assessed. 40 patients underwent PSO with the satellite rod technique (n = 29 at L3, 72.5%; n = 7 at L4, 17.5%). Mean PSO angle was 28.7 ± 7.6°. Two patients (5%) had rod fracture necessitating revision surgery at 32 and 34 months. Three patients (7.5%) developed proximal junctional kyphosis (PJK), but none required revision. No patients developed pseudoarthrosis. Mean preoperative sagittal vertical axis (SVA) was 13.5 ± 7.3 cm, which decreased to 4.8 ± 7.3 cm (P = 0.002) postoperatively. Mean preoperative pelvic incidence-lumbar lordosis (PI-LL) mismatch improved from 35.9 ± 16.6° preoperatively to 11.8 ± 14.6° postoperatively (P < 0.001). Mean preoperative pelvic tilt (PT) improved from 34.7 ± 9.8° preoperatively to 30.1 ± 9.0° (P = 0.026) postoperatively. Mean global alignment and proportion (GAP) score improved from 10 ± 2 preoperatively to 8 ± 2 postoperatively (P < 0.001), which was still considered severely disproportioned. Rod failure rate after PSO with the 4-rod satellite configuration was low. SVA, PI-LL mismatch, and PT significantly improved for these previously-fused patients. Mean GAP score also improved, but was still considered severely disproportioned, likely due to the inability of PSO to correct low LL or PT in previously-fused spines.


Antidiabetic, antihyperlipidaemic, and antioxidant activity of Syzygium densiflorum fruits in streptozotocin and nicotinamide-induced diabetic rats.
Context Syzygium densiflorum Wall. ex Wight & Arn (Myrtaceae) has been traditionally used by local tribes of the Nilgiris, Tamil Nadu, India, for the treatment of diabetes, however, no definitive experimental studies are available. Objective This study investigates the antidiabetic, antihyperlipidaemic and antioxidant activities of ethanol extract of S. densiflorum (EFSD) fruits in streptozotocin (STZ) and nicotinamide (NA)-induced diabetic rats. Materials and methods Acute oral toxicity and oral glucose tolerance were assessed in normal rats. The antidiabetic, antihyperlipidaemic and antioxidant activities were investigated in STZ - NA-induced diabetic rats. Diabetic rats were orally administered with glibenclamide (10 mg/kg b.wt), EFSD (200, 400 and 800 mg/kg b.wt) for 28 d. Further, changes in the blood glucose level (BGL), biochemical parameters, antioxidants were observed and histology of pancreas was performed. Results No toxicity and lethality were observed. Results of the following parameters are represented by treated versus disease control (STZ + NA) groups. BGL (161.33 ± 22.8 versus 476.17 ± 56.58 mg/dl), glycosylated haemoglobin (5.285 ± 0.19 versus 8.05 ± 0.55%), urea (40.32 ± 1.96 versus 75.37 ± 2.91 mg/dl), uric acid (1.2 ± 0.07 versus 2.16 ± 0.05 mg/dl), total cholesterol (89.3 ± 5.14 versus 139.7 ± 5.95 mg/dl) and triglycerides (79.65 ± 2.52 versus 108.9 ± 3.61 mg/dl) were significantly decreased, whereas haemoglobin (11.75 ± 0.73 versus 7.95 ± 0.42 g/dl), high-density lipoprotein cholesterol (14.2 ± 1.11 versus 6.97 ± 0.84 mg/dl), total protein (45%) and liver glycogen (87%) were significantly increased in EFSD-treated diabetic group. Significant changes were observed in the enzymatic and non-enzymatic antioxidants in EFSD-treated groups (p < 0.001). Histopathological examination showed the regeneration of β-cells in Islets of Langerhans. Conclusion This study confirms the antidiabetic, antihyperlipidaemic and antioxidant activities of S. densiflorum fruits.


Three histopathological types of retinoblastoma and their relation to heredity and age of enucleation.
The histopathology of 61 eyes was studied with special attention to the morphology of the retina adjacent to the main tumour. Three retinal types were distinguished. Retina type 1 (RT-1, 28 specimens) contained a single tumour that was sharply demarcated from surrounding normal retina. In retina type 2 (RT-2, 29 specimens) large parts of the retina were affected and the main tumour mass gradually blended with the adjacent pathological retina. Retina type 3 (RT-3, four specimens) was characterised by a retina almost entirely affected by diffuse tumour growth. RT-1 correlated significantly with early enucleation (0-3 years) both in hereditary and non-hereditary cases. RT-2 was seen in eyes enucleated later (2-5 years). The progressing tumour may release growth factors in the intraocular space that stimulate the cells of the adjacent retina and lead to multiple new primary tumours in the adjacent retinal area. RT-3 was only present in non-hereditary cases with late enucleation (at 2-5 years). Hereditary retinoblastoma cases are usually detected early. Therefore in hereditary cases RT-1 is significantly more common than RT-2. In 25 eyes of the 44 patients with unilateral sporadic retinoblastoma, multifocal tumours of the retina were observed. Such cases should not mistakenly be classified as hereditary cases on the basis of the histological pattern of multifocality of the tumour process.


Noninvasive biomarkers in pediatric nonalcoholic fatty liver disease.
Nonalcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver disease worldwide among children and adolescents. It encompasses a spectrum of disease, from its mildest form of isolated steatosis, to nonalcoholic steatohepatitis (NASH) to liver fibrosis and cirrhosis, or end-stage liver disease. The early diagnosis of pediatric NAFLD is crucial in preventing disease progression and in improving outcomes. Currently, liver biopsy is the gold standard for diagnosing NAFLD. However, given its invasive nature, there has been significant interest in developing noninvasive methods that can be used as accurate alternatives. Here, we review noninvasive biomarkers in pediatric NAFLD, focusing primarily on the diagnostic accuracy of various biomarkers as measured by their area under the receiver operating characteristic, sensitivity, and specificity. We examine two major approaches to noninvasive biomarkers in children with NAFLD. First, the biological approach that quantifies serological biomarkers. This includes the study of individual circulating molecules as biomarkers as well as the use of composite algorithms derived from combinations of biomarkers. The second is a more physical approach that examines data measured through imaging techniques as noninvasive biomarkers for pediatric NAFLD. Each of these approaches was applied to children with NAFLD, NASH, and NAFLD with fibrosis. Finally, we suggest possible areas for future research based on current gaps in knowledge.


Expression, function and regulation of mouse cytochrome P450 enzymes: comparison with human P450 enzymes.
The present review focuses on the expression, function and regulation of mouse cytochrome P450 (Cyp) enzymes. Information compiled for mouse Cyp enzymes is compared with data collected for human CYP enzymes. To date, approximately 40 pairs of orthologous mouse-human CYP genes have been identified that encode enzymes performing similar metabolic functions. Recent knowledge concerning the tissue expression of mouse Cyp enzymes from families 1 to 51 is summarized. The catalytic activities of microsomal, mitochondrial and recombinant mouse Cyp enzymes are discussed and their involvement in the metabolism of exogenous and endogenous compounds is highlighted. The role of nuclear receptors, such as the aryl hydrocarbon receptor, constitutive androstane receptor and pregnane X receptor, in regulating the expression of mouse Cyp enzymes is examined. Targeted disruption of selected Cyp genes has generated numerous Cyp null mouse lines used to decipher the role of Cyp enzymes in metabolic, toxicological and biological processes. In conclusion, the laboratory mouse is an indispensable model for exploring human CYP-mediated activities.


KIR2DS1-mediated activation overrides NKG2A-mediated inhibition in HLA-C C2-negative individuals.
NK cell cytotoxicity is controlled through a balance of both activating and inhibitory signals. The HLA specificity of alloreactive NK cells has been previously shown to be controlled by inhibitory killer immunoglobulin-like receptors (KIRs). Alloreactive NK cells lyse targets that lack the HLA ligand for their inhibitory KIR. We have characterized in detail an alloreactive NK clone in which the specificity is controlled by an activating receptor, KIR2DS1. Only target cells expressing the HLA-C group 2 (C2) epitope were lysed by this clone and homozygous C2 targets were lysed more strongly than heterozygous C1/C2 targets. Anti-CD158a (KIR2DS1) blocked lysis of targets confirming KIR2DS1 was responsible. Although this NK clone expressed NKG2A, an inhibitory receptor whose ligand is HLA-E, targets with ligands for both KIR2DS1 and NKG2A were lysed by this clone indicating that the KIR2DS1-mediated activation signal overrides the NKG2A-mediated inhibitory signal. KIR2DS1 activated NK clones in polyclonally expanded NK cultures from a donor that lacked the C2 epitope accounted for approximately 1% of all NK cells. This study highlights a potential role for NK cells controlled by activating KIR in mediating NK alloreactivity.


Ophthalmologic complications after intraoral local anesthesia with articaine.
A series of 14 cases of ophthalmologic complications after intraoral anesthesia of the posterior superior alveolar nerve is presented. The most commonly encountered symptoms were diplopia, mydriasis, palpebral ptosis, and abduction difficulties of the affected eye. In all cases, these effects occurred a few minutes after injection of the anesthetic, followed by complete resolution without sequelae on cessation of the anesthetic effect. The pathogenic mechanism underlying such ophthalmologic disorders is discussed in terms of a possible diffusion of the anesthetic solution toward the orbital region.


Contribution of gold nanoparticles to the catalytic DNA strand displacement in leakage reduction and signal amplification.
A new model using a gold nanoparticle (AuNP)-DNA system to constrain leakage and improve efficiency of catalytic toehold-mediated strand displacement reactions was outlined. A 10-bp spacer on AuNPs and fourfold amount of fuels were determined for good performance of this model with an optimized toehold strategy. After the reaction at 25 °C for 10 h, a 258 pM target could be identified, which is a remarkable improvement compared with the traditional AuNP-DNA system without fuel. Moreover, this model was also studied to differentiate specific single nucleotide polymorphism on target with superior selection factors. This model may help by introducing a proposition of target detection to guide further investigation.


Parity, age at first birth, and risk of death from pancreatic cancer: evidence from a cohort in Taiwan.
This study was undertaken to examine whether there is an association between parity and age at first birth and risk of pancreatic cancer. The study cohort consisted of all women with a record of a first and singleton childbirth in the Birth Registration System between 1978 and 1987. We tracked each woman from the time of their first childbirth and linked their vital status with mortality database. Cox proportional hazard regression models were used to estimate the relative risks (RRs) of death from pancreatic cancer associated with parity and age at first birth. A trend of increasing risk of pancreatic cancer was seen with increasing age at first birth. The adjusted RR was 0.69 (95% confidence interval, 0.49-0.98) for women who born 2 children and 0.64 (95% confidence interval, 0.44-0.93) for women with 3 or more births, respectively, when compared with women who had given birth to only 1 child. There was a significant decreasing trend in the RR of pancreatic cancer with increasing parity. This study provides evidence that reproductive factors (parity and early age at first birth) may confer a protective effect on the risk of pancreatic cancer.


Interferon-alpha in the treatment of Philadelphia-negative chronic myeloproliferative neoplasms. Status and perspectives.
The Philadelphia-negative chronic myeloproliferative neoplasms encompass essential thrombocythemia (ET), polycythemia vera (PV) and primary myelofibrosis (PMF). A major break-through in the understanding of the pathogenesis of these neoplasms occurred in 2005 by the discovery of the JAK2 V617F mutation in the large majority of patients with PV and in half of those with ET and PMF. A number of studies have shown that the "tumor burden" may be monitored at the molecular level in JAK2-positive patients using highly sensitive real-time quantitative PCR. During the last 25 years several studies have shown that interferon-alpha (IFN-alpha) induces complete haematological remissions in a large proportion of the patients. However, its use in clinical practice has unfortunately been limited due to side effects with high drop-out rates in most studies. Recently, IFN-alpha2 has been shown to induce deep molecular remissions and also normalization of the bone marrow in PV, which may be sustained even after discontinuation of IFN-alpha2 therapy. Accordingly, in the coming years we are most likely facing a new era of increasing interest for using IFN-alpha2 in the treatment of patients with PV, ET and the hyperproliferative phase of PMF. This paper reviews the history of IFN - in principle IFN-alpha2 - and its present status in the treatment of PV and related diseases. The role of IFN-alpha2 as immune therapy in the future treatment of CMPNs is highlighted and the rationale for the concept of minimal residual disease and potentially cure after long-term immune therapy with IFN-alpha2 is discussed and foreseen as an achievable goal in the future.


Automated detection of soma location and morphology in neuronal network cultures.
Automated identification of the primary components of a neuron and extraction of its sub-cellular features are essential steps in many quantitative studies of neuronal networks. The focus of this paper is the development of an algorithm for the automated detection of the location and morphology of somas in confocal images of neuronal network cultures. This problem is motivated by applications in high-content screenings (HCS), where the extraction of multiple morphological features of neurons on large data sets is required. Existing algorithms are not very efficient when applied to the analysis of confocal image stacks of neuronal cultures. In addition to the usual difficulties associated with the processing of fluorescent images, these types of stacks contain a small number of images so that only a small number of pixels are available along the z-direction and it is challenging to apply conventional 3D filters. The algorithm we present in this paper applies a number of innovative ideas from the theory of directional multiscale representations and involves the following steps: (i) image segmentation based on support vector machines with specially designed multiscale filters; (ii) soma extraction and separation of contiguous somas, using a combination of level set method and directional multiscale filters. We also present an approach to extract the soma's surface morphology using the 3D shearlet transform. Extensive numerical experiments show that our algorithms are computationally efficient and highly accurate in segmenting the somas and separating contiguous ones. The algorithms presented in this paper will facilitate the development of a high-throughput quantitative platform for the study of neuronal networks for HCS applications.


A Hierarchical Curve-Based Approach to the Analysis of Manifold Data.
One of the data structures generated by medical imaging technology is high resolution point clouds representing anatomical surfaces. Stereophotogrammetry and laser scanning are two widely available sources of this kind of data. A standardised surface representation is required to provide a meaningful correspondence across different images as a basis for statistical analysis. Point locations with anatomical definitions, referred to as landmarks, have been the traditional approach. Landmarks can also be taken as the starting point for more general surface representations, often using templates which are warped on to an observed surface by matching landmark positions and subsequent local adjustment of the surface. The aim of the present paper is to provide a new approach which places anatomical curves at the heart of the surface representation and its analysis. Curves provide intermediate structures which capture the principal features of the manifold (surface) of interest through its ridges and valleys. As landmarks are often available these are used as anchoring points, but surface curvature information is the principal guide in estimating the curve locations. The surface patches between these curves are relatively flat and can be represented in a standardised manner by appropriate surface transects to give a complete surface model. This new approach does not require the use of a template, reference sample or any external information to guide the method and, when compared with a surface based approach, the estimation of curves is shown to have improved performance. In addition, examples involving applications to mussel shells and human faces show that the analysis of curve information can deliver more targeted and effective insight than the use of full surface information.


In vitro chloroquine resistance for Plasmodium vivax isolates from the Western Brazilian Amazon.
Chloroquine (CQ) and primaquine (PQ) are still the drugs of choice to treat Plasmodium vivax malaria in many endemic areas, Brazil included. There is in vivo evidence for the P. vivax resistance to CQ in the Brazilian Amazon, where the increase in the proportion of P. vivax malaria parallels the increase of unusual clinical complications related to this species. In this study, in vitro CQ and mefloquine (MQ)-susceptibility of P. vivax isolates from the Western Brazilian Amazon was tested using the double-site enzyme-linked lactate dehydrogenase immunodetection (DELI) assay. A total of 112 P. vivax isolates were tested in vitro for CQ-susceptibility and out of these 47 were also tested for MQ-susceptibility. The DELI assay was used to detect P. vivax growth at 48-hour short-term culture in isolates with ring stages ranging from 50 to %. Each isolate was tested in triplicate and geometric means of IC50's was obtained. Nineteen isolates were genetically characterized for pvdhfr, pvmrp1, pvmdr1 and pvdhps candidate genes likely related to CQ resistance (10 with IC50<40 nM and 9 with IC50 >100 nM). Twelve out of 112 isolates were considered resistant to CQ, resulting in 10.7% (IC95% 5.0-16.4), while 3 out of 47 (6.4%; IC95% 0.0-12.8) were resistant to MQ. A discrete correlation was observed between IC50's of CQ and MQ (Spearman=0.294; p=0.045). For pvdhps gene, a non-synonymous mutation was found at codon 382 (S→C) in 5/8 CQ-sensitive samples and 1/9 CQ-resistant samples (p=0.027). The other molecular markers were not associated to CQ-susceptibility. In vitro CQ-resistance estimated in this study, estimated by the DELI test, was very similar to that observed in clinical trials, suggesting that in vitro procedures developed by capable local laboratories are useful in the surveillance of CQ-resistance in the Amazon; concurrent Amazon P. vivax strains with both CQ and MQ resistance may be common; and a non-synonymous mutation at pvdhps codon 382 (S→C) was associated to in vitro susceptibility to CQ, needing further studies to be confirmed.


Detection of myocardial inflammation by 18F-FDG-PET/CT in patients with systemic sclerosis without cardiac symptoms: a pilot study.
Primary cardiac manifestation is a common complication of systemic sclerosis (SSc) with poor prognosis. The aim of the current study was to detect potential myocardial inflammation present in asymptomatic SSc patients by 18F-FDG-PET/CT and to investigate its relationship with early signs of myocardial dysfunction as detected by 2D speckle tracking echocardiography (2DSTE). Sixteen consecutive patients with SSc and 9 control patients without clinical evidence of cardiac involvement were enrolled in the study. On 18F-FDG-PET acquired images blood-pool normalised SUV ratio and heterogenity index (HI: standard deviation of SUV divided with mean SUV) were calculated. Within 24 hours all SSc patients underwent 2DSTE strain analysis. Eight of 16 SSc patients were found to be visually PET-positive and showed significantly higher myocardial 18F-FDG SUV ratio (1.78±0.74 vs. 0.98±0.03; p<0.05) and heterogenity index (0.13±0.02 vs. 0.05±0.02; p<0.001) as compared to the control group. FDG-PET/CT derived values did not differ significantly between visually PET-negative (8/16) and control patients (SUV ratio: 0.98±0.05 vs. 0.98±0.03; HI: 0.05±0.01 vs. 0.05±0.02). Global left ventricular longitudinal strain values did not differ significantly between PET-positive and negative patients (17.18±3.49% vs. 17.59±3.65%). Myocardial inflammation, as a potential sign of early cardiac involvement can be detected by 18-FDG-PET/CT in a considerable percentage of systemic sclerosis patients presenting without cardiac symptoms.


Effect of intrathecal baclofen delivered by an implanted programmable pump on health related quality of life in patients with severe spasticity.
To compare clinical effectiveness and health related quality of life in patients with severe spasticity who received intrathecal baclofen or a placebo. In a double blind, randomised, multicentre trial 22 patients were followed up during 13 weeks and subsequently included in a 52 week observational longitudinal study. Patients were those with chronic, disabling spasticity who did not respond to maximum doses of oral baclofen, dantrolene, and tizanidine. After implantation of a programmable pump patients were randomly assigned to placebo or baclofen infusion for 13 weeks. After 13 weeks all patients received baclofen. Clinical efficacy was assessed by the Ashworth scale, spasm score, and self reported pain, and health related quality of life by the sickness impact profile (SIP) and the Hopkins symptom checklist (HSCL). At three months the scores of the placebo and baclofen group differed slightly for the spasm score (effect size=0.20) and substantially for the Ashworth scale (effect size=1.40) and pain score (effect size=0.94); health related quality of life showed no significant differences. Three months after implantation the baclofen group showed a significant, substantial improvement on the SIP "physical health", "mental health", "mobility", and "sleep and rest" subscales and on the HSCL mental health scale; patients receiving placebo showed no change. After one year of baclofen treatment significant (P<0.05) improvement was found on the SIP dimensions "mobility" and "body care and movement" with moderate effect sizes. Improvement on the SIP subscale "physical health" (P<0.05; effect size 0.86), the SIP overall score (without "ambulation"), and the "physical health" and overall scale of the HSCL was also significant, with effect sizes >0.80. Changes in health related behaviour were noted for "sleep and rest" and "recreation and pastimes" (P<0.01, P<0.05; effect size 0.95 and 0.63, respectively). Psychosocial behaviour showed no improvement. Intrathecal baclofen delivered by an implanted, programmable pump resulted in improved self reported quality of life as assessed by the SIP, and HSCL physical health dimensions also suggest improvement.


A double dissociation between sensitivity to changes in object identity and object orientation in the ventral and dorsal visual streams: a human fMRI study.
We used an event-related fMR-adaptation paradigm to investigate changes in BOLD activity in the dorsal and ventral visual streams as a function of object identity and object orientation. Participants viewed successive paired images of real-world, graspable objects, separated by a visual mask. The second image of each pair was either: (i) the same as the first image, (ii) different only in identity, (iii) different only in orientation, or (iv) different in both identity and orientation. A region in the parieto-occipital cortex (dorsal stream) showed a selective increase in BOLD activity with changes in object orientation, but was insensitive to changes in object identity. In contrast, a region in the temporo-occipital cortex (ventral stream) showed a selective increase in activity with changes in identity, but was insensitive to changes in orientation. The differential sensitivity to orientation and identity is consistent with the idea that the dorsal stream plays a critical role in the visual control of object-directed actions while the ventral stream plays a critical role in object perception.


Proteins involved in meiotic recombination: a role in male infertility?
Meiotic recombination results in the formation of crossovers, by which genetic information is exchanged between homologous chromosomes during prophase I of meiosis. Recombination is a complex process involving many proteins. Alterations in the genes involved in recombination may result in infertility. Molecular studies have improved our understanding of the roles and mechanisms of the proteins and protein complexes involved in recombination, some of which have function in mitotic cells as well as meiotic cells. Human gene sequencing studies have been performed for some of these genes and have provided further information on the phenotypes observed in some infertile individuals. However, further studies are needed to help elucidate the particular role(s) of a given protein and to increase our understanding of these protein systems. This review will focus on our current understanding of proteins involved in meiotic recombination from a genomic perspective, summarizing our current understanding of known mutations and single nucleotide polymorphisms that may affect male fertility by altering meiotic recombination.


Effects of intracoronary and intravenous nitroglycerin on coronary collateral function.
Previous studies have demonstrated that nitroglycerin is capable of enhancing coronary collateral function. In the present investigation, the effects of intravenous and intracoronary nitroglycerin were compared to determine whether nitroglycerin augments collateral function by a direct action on the coronary circulation or only by indirect mechanisms. Dogs were studied 2 to 4 weeks after implantation of an ameroid constrictor around the left anterior descending coronary artery. Retrograde flow and peripheral coronary pressure were measured from side arms of a cannula inserted in the left anterior descending coronary artery distal to the ameroid constrictor. Systemic arterial pressure was held constant at approximately 100 mm Hg. Intracoronary nitroglycerin, 0.3 to 100 mug/min, produced dose-dependent increases in retrograde flow but no changes in peripheral coronary pressure. Intravenous nitroglycerin, 10 to 300 mug/min, produced dose-dependent increases in both retrograde flow and pheripheral coronary pressure. Decreasing heart rate from 180 to 60 beats/min increased both retrograde flow and peripheral coronary pressure, but the effect of nitroglycerin on these parameters was not influenced by heart rate. The results indicate that nitroglycerin enhances coronary collateral function by a direct coronary effect and that this direct effect is further augmented by indirect mechanisms. Decreasing heart rate also augments collateral flow, but the effect of nitroglycerin on coronary collateral flow is independent of heart rate.


Small heat shock protein Hsp27 is required for proper heart tube formation.
The small heat shock protein Hsp27 has been shown to be involved in a diverse array of cellular processes, including cellular stress response, protein chaperone activity, regulation of cellular glutathione levels, apoptotic signaling, and regulation of actin polymerization and stability. Furthermore, mutation within Hsp27 has been associated with the human congenital neuropathy Charcot-Marie Tooth (CMT) disease. Hsp27 is known to be expressed in developing embryonic tissues; however, little has been done to determine the endogenous requirement for Hsp27 in developing embryos. In this study, we show that depletion of XHSP27 protein results in a failure of cardiac progenitor fusion resulting in cardia bifida. Furthermore, we demonstrate a concomitant disorganization of actin filament organization and defects in myofibril assembly. Moreover, these defects are not associated with alterations in specification or differentiation. We have thus demonstrated a critical requirement for XHSP27 in developing cardiac and skeletal muscle tissues.


Polypoid lesions of the acalculous gallbladder.
The controversy about gallbladder polyps has centered chiefly around their potential for malignancy. Among 3,525 cholecystectomies done over a 25-year period, there were 26 cases of carcinoma with only 9% not associated with stones. Thirty-nine cases of polypoid lesions of the acalculous gallbladder were investigated. The classic symptoms of pain, flatulence, food intolerance, and nausea were present in the great majority of cases. Most patients, especially those with symptoms of short duration, reported improvement following cholecystectomy.


Potential effects of nutrition-based weight loss therapies in reversing obesity-related breast cancer epigenetic marks.
Obesity is a modifiable risk factor of breast cancer and epigenetic marks were proposed as a relevant mechanistic link. These mechanisms can be remodelled by modifying lifestyle factors and this fact could be useful in the treatment of obesity-related breast cancer. This review aimed to reveal the current evidence on the effects of differences in body composition and lifestyle factors on the risk, treatment, and survival of breast cancer with a focus on the effects of weight loss therapies based on different nutrients, bioactive compounds, and Mediterranean and ketogenic diets to counteract obesity-related breast cancer epigenetic marks. This review was framed on the most relevant and recently published articles and abstracts selected in PubMed using key words related to epigenetics, lifestyle, dietary habits, nutrients, bioactive compounds, ketone bodies, and weight loss treatments in obesity and breast cancer. Several studies have demonstrated that lifestyle interventions, including dietary modifications towards a healthy diet pattern, are effective therapies to prevent the onset of breast cancer and to improve the survival after treatment. These therapies reduce the main factors associated with obesity that are links between adiposity and cancer, including oxidative stress, inflammation and epigenetic mechanisms. However, although sufficient evidence exists regarding the effects of nutrients, dietary patterns, and weight loss therapies to prevent breast cancer or to improve survival, the effects of these strategies on the oncological treatment response were less studied. This review summarises the current scientific evidence regarding these nutritional strategies as adjuvant therapies in the management of obesity-related breast cancer by remodelling epigenetic marks related to carcinogenesis.


The postnatal role of Sox9 in cartilage.
Sox9 is an essential transcription factor for the differentiation of the chondrocytic lineage during embryonic development. To test whether Sox9 continues to play a critical role in cartilaginous tissues in the adult mice, we used an inducible, genetic strategy to disrupt the Sox9 gene postnatally in these tissues. The postnatal inactivation of Sox9 led to stunted growth characterized by decreased proliferation, increased cell death, and dedifferentiation of growth plate chondrocytes. Upon postnatal Sox9 inactivation in the articular cartilage, the sulfated proteoglycan and aggrecan content of the uncalcified cartilage were rapidly depleted and the degradation of aggrecan was accompanied by higher ADAMTS5 immunostaining and increased detection of the aggrecan neoepitope, NITEGE. In spite of the severe loss of Collagen 2a1 mRNA, the Collagen II protein persisted in the articular cartilage, and no histopathological signs of osteoarthritis were observed. The homeostasis of the intervertebral disk (IVD) was dramatically altered upon Sox9 depletion, resulting in disk compression and subsequent degeneration. Inactivation of Sox9 in the IVD markedly reduced the expression of several genes encoding extracellular matrix proteins, as well as some of the enzymes responsible for their posttranslational modification. Furthermore, the loss of Sox9 in the IVD decreased the expression of cytokines, cell-surface receptors, and ion channels, suggesting that Sox9 coordinates a large genetic program that is instrumental for the proper homeostasis of the cells contained in the IVD postnatally. Our results indicate that Sox9 has an essential role in the physiological control of cartilaginous tissues in adult mice. © 2012 American Society for Bone and Mineral Research.


Ensnared by Colorblindness: Discourse on Health Care Disparities.
Race consciousness serves as the foundation for Critical Race Theory (CRT) methodology. Colorblindness minimizes racism as a determinant of outcomes. To achieve the emancipatory intent of CRT and to reduce health care disparities, we must understand: 1) how colorblindness "shows up" when health care professionals aim to promote equity; 2) how their colorblindness informs (and is informed by) clinical practice; and 3) ways to overcome colorblindness through strategies grounded in CRT. We conducted 21 semi-structured interviews with key informants and seven focus groups with personnel employed by a large Minnesota health care system. We coded transcripts inductively and deductively for themes using the constant comparative method. We used a race-conscious approach to examine how respondents' accounts align or diverge from colorblindness. Evading race, respondents considered socioeconomic status, cultural differences, and patients' choices to be the main contributors to health disparities. Few criticized the behavior of coworkers or that of the organization or acknowledged structural racism. Respondents strongly believed that all patients were treated equally by providers and staff, in part due to race-neutral care processes and guidelines. Respondents also used several semantic moves common to colorblindness to refute suggestions of racial inequality. Colorblindness upholds the racial status quo and inhibits efforts to promote health equity. Drawing on CRT to guide them, health care leaders will need to develop strategies to counter personnel's tendency to focus on axes of inequality other than race, to decontextualize patients' health behaviors and choices, and to depend heavily on race-neutral care processes to produce equitable outcomes.


Vibrationally excited NO tagging by NO(A²∑⁺) fluorescence and quenching for simultaneous velocimetry and thermometry in gaseous flows.
We present measurements demonstrating simultaneous determination of velocity and temperature using a variant of the Vibrationally Excited Nitric Oxide Monitoring (VENOM) technique that does not employ NO2. The variant is based on tagging by electronic excitation of NO in the A²∑(1/2)⁺ (v'=0)←X2Π1/2(v''=0) band and subsequent formation of vibrationally excited NO(X2Π) by spontaneous emission and collisional quenching. Sequential planar laser-induced fluorescence imaging of the nascent NO(X2Π, v''=1) was used to obtain spatially resolved average streamwise velocity and rotational/translational temperature. The temperature determination using this approach extends the applicability of the VENOM technique to low-density, high-speed flows, where slow thermalization of the tagged molecules represents a limiting factor.


Persistent nitrogen limitation of stream biofilm communities along climate gradients in the Arctic.
Climate change is rapidly reshaping Arctic landscapes through shifts in vegetation cover and productivity, soil resource mobilization, and hydrological regimes. The implications of these changes for stream ecosystems and food webs is unclear and will depend largely on microbial biofilm responses to concurrent shifts in temperature, light, and resource supply from land. To study those responses, we used nutrient diffusing substrates to manipulate resource supply to biofilm communities along regional gradients in stream temperature, riparian shading, and dissolved organic carbon (DOC) loading in Arctic Sweden. We found strong nitrogen (N) limitation across this gradient for gross primary production, community respiration and chlorophyll-a accumulation. For unamended biofilms, activity and biomass accrual were not closely related to any single physical or chemical driver across this region. However, the magnitude of biofilm response to N addition was: in tundra streams, biofilm response was constrained by thermal regimes, whereas variation in light availability regulated this response in birch and coniferous forest streams. Furthermore, heterotrophic responses to experimental N addition increased across the region with greater stream water concentrations of DOC relative to inorganic N. Thus, future shifts in resource supply to these ecosystems are likely to interact with other concurrent environmental changes to regulate stream productivity. Indeed, our results suggest that in the absence of increased nutrient inputs, Arctic streams will be less sensitive to future changes in other habitat variables such as temperature and DOC loading.


National Hospital Discharge Survey: 2002 annual summary with detailed diagnosis and procedure data.
This report presents 2002 national estimates and selected trend data on the use of non-Federal short-stay hospitals in the United States. Estimates are provided by selected patient and hospital characteristics, diagnoses, and surgical and nonsurgical procedures performed. Estimates of diagnoses and procedures are presented according to International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) codes. The estimates are based on data collected through the National Hospital Discharge Survey (NHDS). The survey has been conducted annually since 1965. In 2002, data were collected for approximately 327,000 discharges. Of the 474 eligible non-Federal short-stay hospitals in the sample, 445 (94 percent) responded to the survey. An estimated 33.7 million inpatients were discharged from non-Federal short-stay hospitals in 2002. They used 164.2 million days of care and had an average length of stay of 4.9 days. Common first-listed discharge diagnoses included delivery, ischemic heart disease, psychoses, pneumonia, and malignant neoplasms. Inpatients had 6.8 million cardiovascular procedures and 6.6 million obstetric procedures. Males had higher rates for cardiac procedures such as cardiac catheterization and coronary artery bypass graft, but males and females had similar rates of pacemaker procedures. The number and rate of all cesarean deliveries, primary and repeat, rose from 1995 to 2002; the rate of vaginal birth after cesarean delivery dropped from 35.5 in 1995 to 15.8 in 2002.


Long-term analysis of GOME in-flight calibration parameters and instrument degradation.
Since 1995, the Global Ozone Monitoring Experiment (GOME) has measured solar and backscattered spectra in the ultraviolet and visible wavelength range. Now, the extensive data set of the most important calibration parameters has been investigated thoroughly in order to analyze the long-term stability and performance of the instrument. This study focuses on GOME in-flight calibration and degradation for the solar path. Monitoring the sensor degradation yields an intensity decrease of 70% to 90% in 240-316 nm and 35% to 65% in 311-415 nm. The spectral calibration is very stable over the whole period, although a very complex interaction between predisperser temperature and wavelength was found. The leakage current and the pixel-to-pixel gain increased significantly during the mission, which requires an accurate correction of the measured radiance and irradiance signals using proper calibration parameters. Finally, several outliers in the data sets can be directly assigned to instrument and satellite anomalies.


Serum HCoV-spike specific antibodies do not protect against subsequent SARS-CoV-2 infection in children and adolescents.
SARS-CoV-2 infections in children are generally asymptomatic or mild and rarely progress to severe disease and hospitalization. Why this is so remains unclear. Here we explore the potential for protection due to pre-existing cross-reactive seasonal coronavirus antibodies and compare the rate of antibody decline for nucleocapsid and spike protein in serum and oral fluid against SARS-CoV-2 within the pediatric population. No differences in seasonal coronaviruses antibody concentrations were found at baseline between cases and controls, suggesting no protective effect from pre-existing immunity against seasonal coronaviruses. Antibodies against seasonal betacoronaviruses were boosted in response to SARS-CoV-2 infection. In serum, anti-nucleocapsid antibodies fell below the threshold of positivity more quickly than anti-spike protein antibodies. These findings add to our understanding of protection against infection with SARS-CoV-2 within the pediatric population, which is important when considering pediatric SARS-CoV-2 immunization policies.


Hydration of the dTn.dAn x dTn parallel triple helix: a Fourier transform infrared and gravimetric study correlated with molecular dynamics simulations.
We present a comparative analysis of the water organization around the dTn.dAn x dTn triple helix and the Watson-Crick double helix dTn.dAn respectively by means of gravimetric measurements, infrared spectroscopy and molecular dynamics simulations. The hydration per nucleotide determined by gravimetric and spectroscopic methods correlated with the molecular dynamics simulations shows that at high relative humidity (98% RH) the triple helix is less solvated than the duplex (17 +/- 2 water molecules per nucleotide instead of 21 +/-1). The experimental desorption curves are different for both structures and indicate that below 81% RH the triplex becomes more hydrated than the duplex. At this RH the FTIR spectra show the emergence of N-type sugars in the adenosine strand of the triplex. When the third strand is bound in the major groove of the Watson-Crick duplex molecular dynamics simulations show the formation of a spine of water molecules between the two thymidine strands.


Outcomes of Patients Who Undergo Cardiac Surgical Procedures After Liver Transplantation.
There is a paucity of information available regarding the impact of cardiac surgical procedures on patients who have undergone previous liver transplantation. The primary purpose of this study was to ascertain the survival rate and predictors of death in this specific patient population. This retrospective cohort study consisted of a consecutive series of patients with a functioning liver allograft who subsequently underwent cardiac surgical procedures between January 1991 and December 2012. The optimal Model for End-Stage Liver Disease (MELD) score for predicting late death was identified using receiver operating characteristic curve analysis. Risk of postoperative death was determined by parametric hazard analysis. Between January 1991 and December 2012, 43 patients (median age, 60 years) underwent cardiac surgical procedures after liver transplantation. The median interval between liver transplant and cardiac operation was 63 months (range, 1.1 to 217 months). Three operative deaths and 24 late deaths occurred. Receiver operating characteristic curve analysis identified the optimal preoperative and postoperative MELD score cut points for predicting late death as greater than 13.8 (area under the curve = 0.674) and greater than 17 (area under the curve = 0.633), respectively. Patients with a preoperative MELD score of 13.8 or less had significantly greater survival rates than those with a MELD score greater than 13.8 (p = 0.028); patients with a postoperative MELD score of 17 of less had significantly greater survival rates than those with a MELD score greater than 17 (p < 0.001). Multivariate parametric hazard analysis identified postoperative peak creatinine level as a statistically significant predictor of death (relative risk, 1.8; p = 0.01). The 1-, 5-, and 10-year Kaplan-Meier survival rates were 90%, 51%, and 35%, respectively; postoperative mortality rates followed a constant phase model with a hazard of death of 10% per year. Cardiac surgical procedures can be performed with acceptable short-term and long-term outcomes in liver transplant recipients. Elevated preoperative and postoperative MELD scores and postoperative peak creatinine level may portend death in this cohort. There is a constant hazard of death of 10% per year.


MYB-bHLH-TTG1 Regulates Arabidopsis Seed Coat Biosynthesis Pathways Directly and Indirectly via Multiple Tiers of Transcription Factors.
MYB-bHLH-WDR (MBW) transcription factor (TF) complexes regulate Arabidopsis seed coat development including mucilage and tannin biosynthesis. The R2R3 MYBs MYB5, MYB23 and TRANSPARENT TESTA2 (TT2) participate in the MBW complexes with the WD-repeat protein TRANSPARENT TESTA GLABRA1 (TTG1). These complexes regulate GLABRA2 (GL2) and TTG2 expression in developing seeds. Microarray transcriptome analysis of ttg1-1- and wild-type (Ler) developing seeds identified 246 TTG1-regulated genes, which include all known metabolic genes of the tannin biosynthetic pathway. The first detailed TTG1-dependent metabolic pathways could be proposed for the biosynthesis of mucilage, jasmonic acid (JA) and cuticle including wax ester in developing seeds. We also assigned many known and previously uncharacterized genes to the activation/inactivation of hormones, plant immunity and nutrient transport. The promoters of six cuticle pathway genes were active in developing seeds. Expression of 11 genes was determined in the developing seeds of the combinatorial mutants of MYB5, MYB23 and TT2, and in the combinatorial mutants of GL2, HOMEODOMAIN GLABROUS2 (HDG2) and TTG2. These six TFs positively co-regulated the expression of four repressor genes while three of the six TFs repressed the wax biosynthesis genes examined, suggesting that the three TFs upregulate the expression of these repressor genes, which, in turn, repress the wax biosynthesis genes. Chromatin immunoprecipitation analysis identified 21 genes directly regulated by MYB5 including GL2, HDG2, TTG2, four repressor genes and various metabolic genes. We propose a multi-tiered regulatory mechanism by which MBWs regulate tannin, mucilage, JA and cuticle biosynthetic pathways.


Gastrin-releasing peptide mediates light-like resetting of the suprachiasmatic nucleus circadian pacemaker through cAMP response element-binding protein and Per1 activation.
Circadian rhythmicity in the primary mammalian circadian pacemaker, the suprachiasmatic nucleus (SCN) of the hypothalamus, is maintained by transcriptional and translational feedback loops among circadian clock genes. Photic resetting of the SCN pacemaker involves induction of the clock genes Period1 (Per1) and Period2 (Per2) and communication among distinct cell populations. Gastrin-releasing peptide (GRP) is localized to the SCN ventral retinorecipient zone, from where it may communicate photic resetting signals within the SCN network. Here, we tested the putative role of GRP as an intra-SCN light signal at the behavioral and cellular levels, and we also tested whether GRP actions are dependent on activation of the cAMP response element-binding protein (CREB) pathway and Per1. In vivo microinjections of GRP to the SCN regions of Per1::green fluorescent protein (GFP) mice during the late night induced Per1::GFP throughout the SCN, including a limited population of arginine vasopressin-immunoreactive (AVP-IR) neurons. Blocking spike-mediated communication with tetrodotoxin did not disrupt overall Per1::GFP induction but did reduce induction within AVP-IR neurons. In vitro GRP application resulted in persistent increases in the spike frequency of Per1::GFP-induced neurons. Blocking endogenous Per1 with antisense oligodeoxynucleotides inhibited GRP-induced increases in spike frequency. Furthermore, inhibition of CREB-mediated gene activation with decoy oligonucleotides blocked GRP-induced phase shifts of PER2::luciferase rhythms in SCN slices. Altogether, these results indicate that GRP communicates phase resetting signals within the SCN network via both spike-dependent and spike-independent mechanisms, and that activation of the CREB pathway and Per1 are key steps in mediating downstream events in GRP resetting of SCN neurons.


Glucocorticoids antagonize estrogens by glucocorticoid receptor-mediated activation of estrogen sulfotransferase.
Glucocorticoids and estrogens are two classes of steroid hormones that have essential but distinct physiologic functions. Estrogens also represent a risk factor for breast cancer. It has been suggested that glucocorticoids can attenuate estrogen responses, but the mechanism by which glucocorticoids inhibit estrogenic activity is unknown. In this study, we show that activation of glucocorticoid receptor (GR) by dexamethasone (DEX) induced the expression and activity of estrogen sulfotransferase (SULT1E1 or EST), an enzyme important for the metabolic deactivation of estrogens, because sulfonated estrogens fail to activate the estrogen receptor. Treatment with DEX lowered circulating estrogens, compromised uterine estrogen responses, and inhibited estrogen-dependent breast cancer growth in vitro and in a xenograft model. We further showed that the mouse and human SULT1E1 genes are transcriptional targets of GR and deletion of Sult1e1/Est in mice abolished the DEX effect on estrogen responses. These findings have revealed a novel nuclear receptor-mediated and metabolism-based mechanism of estrogen deprivation, which may have implications in therapeutic development for breast cancers. Because glucocorticoids and estrogens are widely prescribed drugs, our results also urge caution in avoiding glucocorticoid-estrogen interactions in patients.


A case of obesity, diabetes and hypertension treated with very low calorie diet (VLCD) followed by successful pregnancy with intrauterine insemination (IUI).
The patient was a 32-year-old obese woman with a history of type 2 diabetes and hypertension for 6 years. Although she was treated with antihypertensive agents and intensive insulin therapy, her hyperglycemia was difficult to control. She wanted to have a baby but pregnancy was not recommended because her diabetes was under poor control and the use of antihypertensive medication. To achieve good control of obesity, diabetes and hypertension, she was admitted to our clinical department for weight reduction using very low calorie diet (VLCD). During VLCD she had a 19.8 kg reduction in body weight and her blood glucose and blood pressure were in good control without the use of drugs. Five months later, she became pregnant after the fourth trial of intrauterine insemination (IUI) and gave birth to a female baby under insulin therapy. This is the first report that showed the usefulness of VLCD for prepregnant control of glucose metabolism and blood pressure in an obese hypertensive patient with type 2 diabetes mellitus.


Adenosine 3',5'-monophosphate primes the secretion of follicle-stimulating hormone.
We investigated whether cAMP acts as a mediator for LHRH in either its immediate FSH release action or its self-priming action. Pituitary pieces from cyclic female rats were superfused in vitro in the presence of Bu2cAMP, 8-bromo-cAMP, or forskolin or used as controls. For pituitary pieces from proestrous rats, the first significant increase in the baseline FSH secretion rate occurred after approximately 90 min of exposure to elevated cAMP resulting from forskolin treatment. By comparison, in the same system LHRH caused a 3-fold increase in FSH secretion during a 10-min exposure to the peptide. In contrast to its ineffectiveness as a secretagogue, cAMP elevation resulted in a several-fold augmentation of both LHRH- and elevated K+-stimulated FSH secretion from pituitary pieces from proestrous, but not estrous, rats; for these experiments, superfusion with a cAMP analog or forskolin for varying times preceded a 10-min pulse of either 8 nM LHRH or 47 mM K+. Augmentation of K+-stimulated secretion was evident after 30 min of cAMP elevation. Priming of LHRH-stimulated FSH secretion required 30-90 min of pretreatment with cAMP; longer exposures to cAMP analogs or forskolin were coincident with greater potentiation. Cycloheximide prevented Bu2cAMP augmentation of LHRH-stimulated FSH secretion. These data show that cAMP does not mimic the FSH release action of LHRH, but does augment LHRH- or K+-stimulated FSH secretion with characteristics that lead us to suggest that cAMP mediates, at least in part, the self-priming function of LHRH.


Rat liver carbonic anhydrase I is not sexually dimorphic but is estrogen repressible.
Carbonic anhydrase (CA) isozymes CAII and CAIII are known to exhibit sexual dimorphism in rat liver, and the levels of these isozymes are affected by sex hormones. In this paper we show that the isozyme CAI is present at low levels in rat liver, with no difference in concentration between male and female rats. Estrogen and diethylstilbestrol reduce CAI levels in both sexes.


Influence of phenylacetic acid pulses on anaerobic digestion performance and archaeal community structure in WWTP sewage sludge digesters.
The effect of phenylacetic acid (PAA) pulses on anaerobic digestion (AD) performance and archaeal community structure was evaluated in anaerobic digesters treating sewage sludge from a wastewater treatment plant (WWTP). Four pilot-scale continuous stirred tank reactors were set up at a full-scale municipal WWTP in Santiago de Chile, and fed with either primary or mixed sewage sludge. AD performance was evaluated by volatile fatty acid (VFA) and biogas production monitoring. Archaeal community structure was characterized by 16S rRNA denaturing gradient gel electrophoresis and band sequencing. In the primary sludge digester, a single PAA pulse at 200 mg L(-1) was sufficient to affect AD performance and archaeal community structure, resulting in long-term VFA accumulation, reduced biogas production and community shift from dominant acetoclastic (Methanosaeta concilii) to hydrogenotrophic (Methanospirillum hungatei) methanogens. By contrast, AD performance and archaeal community structure in the mixed sludge digester were stable and resistant to repeated PAA pulses at 200 and 600 mg L(-1). This work demonstrated that the effect of PAA pulses on methanogenic activity and archaeal community structure differed according to AD substrate, and suggests that better insights of the correlations between archaeal population dynamics and functional performance could help to better face toxic shocks in AD.


Targeting Striatal Glutamate and Phosphodiesterases to Control L-DOPA-Induced Dyskinesia.
A large body of work during the past several decades has been focused on therapeutic strategies to control L-DOPA-induced dyskinesias (LIDs), common motor complications of long-term L-DOPA therapy in Parkinson's disease (PD). Yet, LIDs remain a clinical challenge for the management of patients with advanced disease. Glutamatergic dysregulation of striatal projection neurons (SPNs) appears to be a key contributor to altered motor responses to L-DOPA. Targeting striatal hyperactivity at the glutamatergic neurotransmission level led to significant preclinical and clinical trials of a variety of antiglutamatergic agents. In fact, the only FDA-approved treatment for LIDs is amantadine, a drug with NMDAR antagonistic actions. Still, novel agents with improved pharmacological profiles are needed for LID therapy. Recently other therapeutic targets to reduce dysregulated SPN activity at the signal transduction level have emerged. In particular, mechanisms regulating the levels of cyclic nucleotides play a major role in the transduction of dopamine signals in SPNs. The phosphodiesterases (PDEs), a large family of enzymes that degrade cyclic nucleotides in a specific manner, are of special interest. We will review the research for antiglutamatergic and PDE inhibition strategies in view of the future development of novel LID therapies.


[Pulmonary hemangiopericytoma. Apropos of a case and review of the literature].
Hemangiopericytoma is a rare vascular tumor of pericyte origin and of variable but always preoccupying potential since falling within the category of sarcomas. A thoracic localization is rare and primary pulmonary parenchyma lesions even more so, only 87 cases being reported in the literature. Radiological diagnostic criteria are of weak specificity, diagnosis being provided by thin section CT scan imaging with contrast. The differential diagnosis of these vascular tumors includes arteriovenous fistula, pulmonary varices, other vascular tumors or even organized bronchomucoceles. The value of CT scan puncture is obviously limited. The course is unpredictable since even histopathology findings fail to provide definitive data.


Trunk lean mass and its association with 4 different measures of thoracic kyphosis in older community dwelling persons.
The causes of age-related hyperkyphosis (HK) include osteoporosis, but only 1/3 of those most severely affected have vertebral fractures, suggesting that there are other important, and potentially modifiable causes. We hypothesized that muscle mass and quality may be important determinants of kyphosis in older persons. We recruited 72 persons >65 years to participate in a prospective study designed to evaluate kyphosis and fall risk. At the baseline visit, participants had their body composition measures completed using Dual Energy X-ray Absorptiometry (DXA). They had kyphosis measured in either the standing [S] or lying [L] position: 1) Cobb angle from DXA [L]; 2) Debrunner kyphometer [S]; 3) architect's flexicurve ruler [S]; and 4) blocks method [L]. Multivariable linear/logistic regression analyses were done to assess the association between each body composition and 4 kyphosis measures. Women (n = 52) were an average age of 76.8 (SD 6.7) and men 80.5 (SD 7.8) years. They reported overall good/excellent health (93%), the average body mass index was 25.3 (SD 4.6) and 35% reported a fall in the past year. Using published cut-offs, about 20-30% were determined to have HK. For the standing assessments of kyphosis only, after adjusting for age, sex, weight and hip BMD, persons with lower TLM were more likely to be hyperkyphotic. Lower TLM is associated with HK in older persons. The results were stronger when standing measures of kyphosis were used, suggesting that the effects of muscle on thoracic kyphosis are best appreciated under spinal loading conditions.


Polymerase chain reaction analysis for viruses in paraffin-embedded myocardium from dogs with dilated cardiomyopathy or myocarditis.
To perform polymerase chain reaction (PCR) analysis on paraffin-embedded myocardium from dogs with dilated cardiomyopathy (DCM) and dogs with myocarditis to screen for canine parvovirus, adenovirus types 1 and 2, and herpesvirus. Myocardial specimens from 18 dogs with an antemortem diagnosis of DCM and 9 dogs with a histopathologic diagnosis of myocarditis were evaluated. Paraffin-embedded myocardial specimens were screened for viral genome by PCR analysis. Positive-control specimens were developed from cell cultures as well as paraffin-embedded tissue specimens from dogs with clinical and histopathologic diagnoses of viral infection with canine parvovirus, adenovirus types 1 and 2, and herpesvirus. The histologic characteristics of all myocardial specimens were classified regarding extent, location, and type of inflammation and fibrosis. Canine adenovirus type 1 was amplified from 1 specimen from a dog with DCM. Canine parvovirus, adenovirus type 2, and herpesvirus were not amplified from any myocardial specimens. Histologic analysis of specimens from dogs with DCM revealed variable amounts of fibrosis; myocardial inflammation was observed in 1 affected dog. Histopathologic analysis of specimens from dogs with myocarditis disclosed variable degrees of inflammation and fibrosis. Viral agents canine parvovirus, adenovirus types 1 and 2, and herpesvirus are not commonly associated with DCM or active myocarditis in dogs. Additional studies evaluating for nucleic acid from viruses that less commonly affect dogs or different types of infectious agents may be warranted to gain insight into the cause of DCM and myocarditis in dogs.


Immunogenic, non-infectious polio subviral particles synthesized in Saccharomyces cerevisiae.
Polioviral genes coding for P1, the precursor for the structural proteins, and 3CD, the viral protease, were cloned in a Saccharomyces cerevisiae inducible expression system. N-antigenic empty capsids could be isolated from the yeast cell extract provided that pirodavir, a capsid-binding compound and capsid stabilizer, was added during the induction period and during purification. Purification was by immunoaffinity chromatography. The purified empty capsids had the same immunogenicity as poliovirus virions. The techniques described might be useful for the production of new non-infectious vaccines.


Permeabilizing action of polyethyleneimine on Salmonella typhimurium involves disruption of the outer membrane and interactions with lipopolysaccharide.
Polyethyleneimine (PEI), a polycationic polymer substance used in various bioprocesses as a flocculating agent and to immobilize enzymes, was recently shown to make Gram-negative bacteria permeable to hydrophobic antibiotics and to detergents. Because this suggests impairment of the protective function of the outer membrane (OM), the effect of PEI on the ultrastructure of Salmonella typhimurium was investigated. Massive alterations in the OM of PEI-treated and thin-sectioned bacteria were observed by electron microscopy. Vesicular structures were seen on the surface of the OM, but no liberation of the membrane or its fragments was evident. Since a potential mechanism for the action of PEI could be its binding to anionic LPSs on the OM surface, the interaction of PEI with isolated LPSs was assayed in vitro. The solubility of smooth-type LPSs of Salmonella, regardless of the sugar composition of their O-specific chains, was not affected by PEI, nor was that of Ra-LPS (lacking O-specific chains but having a complete core oligosaccharide). PEI strongly decreased the solubility of rough-type LPSs of the chemotypes Rb2 and Re, whereas it had only a weak effect on the abnormally cationic Rb2-type pmrA mutant LPS, suggesting that the negative charge to mass ratio of LPS plays a critical role in the interaction.


Validation of a new 96-well plate spectrophotometric method for the quantification of compound 48/80 associated with particles.
A new, simple, inexpensive, and rapid 96-well plate UV spectrophotometric method was developed and validated for the quantification of compound 48/80 (C48/80) associated with particles. C48/80 was quantified at 570 nm after reaction with acetaldehyde and sodium nitroprusside in an alkaline solution (pH 9.6). The method was validated according to the recommendations of the ICH Guidelines for specificity, linearity, range, accuracy, precision, and detection and quantification limits (DL and QL). All the validation parameters were assessed in three different solvents, i.e., deionized water, blank matrix of chitosan nanoparticles, and blank matrix of chitosan/alginate nanoparticles. The method was found to be linear in the concentration range of 5 to 160 μg/ml (R(2)>0.9994). Intraday and interday precision was adequate, with relative standard deviation lower than those given by the Horwitz equation. The mean recoveries of C48/80 from spiked samples ranged between 98.1% and 105.9% for calibration curves done with the blank matrices and between 89.3% and 103.3% for calibration curves done with water, respectively. The DL were lower than 1.01 μg/ml and the QL were lower than 3.30 μg/ml. The results showed that the developed method is sensitive, linear, precise, and accurate for its intended use, with the additional advantages of being cost-effective and time-effective, allowing the use of small-volume samples, and the simultaneous analysis of a large number of samples. The proposed method was already successfully applied to evaluate the loading efficacy of C48/80 chitosan-based nanoparticles and can be easily applied during the development of other C48/80-based formulations.


A rapid enzyme immunofiltration technique using monoclonal antibodies to serotype herpes simplex virus.
A rapid enzyme immunofiltration technique using monoclonal antibodies to serotype herpes simplex virus is described. It requires only a single tube culture showing viral cytopathology and can accommodate multiple specimens in a single assay. The monoclonal antibodies confer absolute specificity and the use of horseradish peroxidase-conjugated staphylococcal protein A or antiglobulin permits easy visual interpretation of the results following the 2-3 hour assay. Although it is possible that an occasional wild strain of HSV might escape recognition by monoclonal antibody, this potential problem has not been observed among the more than 500 clinical isolates tested to date, all of which have yielded an unequivocal result.


Reactive oxygen species-independent G1 arrest induced by evening primrose extract in Ehrlich ascites tumor cells.
We previously demonstrated that evening primrose extract (EPE) induced apoptosis in Ehrlich ascites tumor cells (EATC), and this effect was specific on tumor cells. Furthermore, our results demonstrated that EPE exposure elicited a rapid increase in the activity of superoxide dismutase and intracellular peroxides levels. These changes caused translocation of Bax to mitochondria and a subsequent release of mitochondrial cytochrome c. However, no activation of caspase-3 was observed in EPE-treated EATC. On the other hand, apoptosis-inducing factor (AIF) was translocated from mitochondria to nuclei. The EPE-induced translocation of AIF was suppressed with the addition of catalase, suggesting that the rapid intracellular peroxide levels after addition of EPE triggers off induction of apoptosis, which is AIF-mediated and caspase-independent. In this study, we have shown that EPE elicited a dose-dependent accumulation of cells in the G1 phase and inhibited DNA synthesis. Our results also demonstrated that cell cycle arrest and inhibition of proliferation in EATC by EPE are associated with decreased Rb phosphorylation. Furthermore, inhibitions of Rb phosphorylation and DNA synthesis by EPE were not suppressed with the addition of catalase. The present study suggests that intracellular peroxides, which trigger off induction of apoptosis, are not the trigger of EPE-induced G1 arrest in cell cycle.


Smoking and subarachnoid haemorrhage.
An enquiry was made into the smoking habits of 199 patients who had survived a subarachnoid haemorrhage (SAH). Information was obtained from 189, giving a 95% response rate. Those with a cerebral aneurysm had on average smoked considerably more cigarettes daily than those with arteriovenous malformations (AVM) or with normal angiographic findings (NAF) whose smoking habits were similar to those of the UK population. SAH due to cerebral aneurysm should be considered another smoking-related disease.


Ameliorative effects of Bacopa monniera on lead-induced oxidative stress in different regions of rat brain.
Bacopa monniera is a rejuvenating herb for brain cells enhancing learning and cognitive ability. In the present investigation, the ameliorative effects of Bacopa monniera were examined against lead-induced oxidative stress in different regions of rat brain. Male rats were divided into five groups: control (1000 ppm sodium acetate) and exposed (1000 ppm lead acetate) for 4 weeks; DMSA (Meso-2,3-Dimercaptosuccinic acid)-treated (90 mg/kg body weight/day); Bacopa monniera-treated (BM) (10 mg/kg body weight/day) and a combination of BM + DMSA for seven consecutive days after 4 weeks of lead exposure. After treatment, the whole brain was isolated by sacrificing rats and four regions were separated namely cerebellum, hippocampus, frontal cortex and brain stem. Results indicated a significant (p < 0.05) increase in reactive oxygen species (ROS), lipid peroxidation products (LPP) and total protein carbonyl content (TPCC) in association with tissue metal content in all the four regions of brain for exposed group compared with their respective controls. However, the lead-induced ROS, LPP, TPCC and tissue metal content were lowered on treatment with Bacopa monniera, almost reaching the control group values in all the above brain regions compared to DMSA and a combination therapy. Results suggest that Bacopa monniera can mitigate the lead induced-oxidative stress tissue specifically by pharmacologic interventions which encompass both chelation as well as antioxidant functions.


Six-Membered Aromatic Nitrogen Heterocyclic Anti-Tumor Agents: Synthesis and Applications.
Cancer stands as a serious malady, posing substantial risks to human well-being and survival. This underscores the paramount necessity to explore and investigate novel antitumor medications. Nitrogen-containing compounds, especially those derived from natural sources, form a highly significant category of antitumor agents. Among these, antitumor agents with six-membered aromatic nitrogen heterocycles have consistently attracted the attention of chemists and pharmacologists. Accordingly, we present a comprehensive summary of synthetic strategies and clinical implications of these compounds in this review. This entails an in-depth analysis of synthesis pathways for pyridine, quinoline, pyrimidine, and quinazoline. Additionally, we explore the historical progression, targets, mechanisms of action, and clinical effectiveness of small molecule inhibitors possessing these structural features.


A short update on the structure of drug binding sites on neurotransmitter transporters.
The dopamine (DAT), noradrenalin (NET) and serotonin (SERT) transporters are molecular targets for different classes of psychotropic drugs. Cocaine and the SSRI (S)-citalopram block neurotransmitter reuptake competitively, but while cocaine is a non-selective reuptake inhibitor, (S)-citalopram is a selective SERT inhibitor. Here we present comparisons of the binding sites and the electrostatic potential surfaces (EPS) of DAT, NET and SERT homology models based on two different LeuTAa templates; with a substrate (leucine) in an occluded conformation (PDB id 2a65), and with an inhibitor (tryptophan) in an open-to-out conformation (PDB id 3f3a). In the occluded homology models, two conserved aromatic amino acids (tyrosine and phenylalanine) formed a gate between the putative binding pockets, and this contact was interrupted in the open to out conformation. The EPS of DAT and NET were generally negative in the vestibular area, whereas the EPS of the vestibular area of SERT was more neutral. The findings presented here contribute as an update on the structure of the binding sites of DAT, NET and SERT. The updated models, which have larger ligand binding site areas than models based on other templates, may serve as improved tools for virtual ligand screening.


IκBα is not required for axon initial segment assembly.
The inhibitor of NF-κB alpha (IκBα) protein is an important regulator of the transcription factor NF-κB. In neurons, IκBα has been shown to play a role in neurite outgrowth and cell survival. Recently, a phosphorylated form of IκBα (pIκBα Ser32/36) was reported to be highly enriched at the axon initial segment (AIS) and was proposed to function upstream of ankyrinG in AIS assembly, including ion channel recruitment. However, we report here that the AIS clustering of ankyrinG and Na(+) channels in the brains of IκBα knockout (Nfkbia(-/-)) mice is comparable to that in wild-type littermates. Furthermore, we found that multiple phospho-specific antibodies against pIκBα Ser32/36 non-specifically label AIS in Nfkbia(-/-) cortex and AIS in dissociated Nfkbia(-/-) hippocampal neurons. With the exception of ankyrinG, shRNA-mediated knockdown of known AIS proteins in cultured hippocampal neurons did not eliminate the AIS labeling with pIκBα antibodies. Instead, the pIκBα antibodies cross-react with a phosphorylated epitope of a protein associated with the microtubule-based AIS cytoskeleton that is not integrated into the AIS membrane complex organized by ankyrinG. Our results indicate that pIκBα is neither enriched at the AIS nor required for AIS assembly.


A new approach to the inflammatory/autoimmune diseases.
Extracellular adenosine 5 -triphosphate (eATP) is an important mediator of cell-to-cell interactions in the nervous, vascular and immune system. Its low release by different cells, as mast cells, platelets, red blood cells, T cells and also by nerve terminals, requires different mechanisms and especially occurs in physiologic conditions. However, in pathologic conditions, as inflammation, eATP can highly increase, following its release by damaged cells. Elevated levels of eATP provoke the activation of some purinergic receptors, mainly the P2XR, which are located on mononuclear phagocytes, T cells and endothelial cells. The activation by eATP of inflammatory/immune cells leads to their release of some inflammatory mediators, such as the cytokines IL-1beta and TNFalpha. These cytokines are able to further activate other cells, as endothelial cells, favouring their increased expression of adhesion molecules; such process enhances circulating cell recruitment to the inflamed tissue. The blockade of the purinergic-mediated activation of the inflammatory/immune cells might represent a useful tool to reduce the spreading of the inflammatory/immune response. This review will summarize the beneficial effects of the use of periodate oxidized ATP (oATP), an inhibitor of P2XR, in the treatment of some experimental models of inflammatory/immune diseases. The article is a short review of recent patents related to the anti-inflammatory/analgesic/antiangiogenic effects of oATP and to its role in the autoimmune diseases.


Noisy splicing drives mRNA isoform diversity in human cells.
While the majority of multiexonic human genes show some evidence of alternative splicing, it is unclear what fraction of observed splice forms is functionally relevant. In this study, we examine the extent of alternative splicing in human cells using deep RNA sequencing and de novo identification of splice junctions. We demonstrate the existence of a large class of low abundance isoforms, encompassing approximately 150,000 previously unannotated splice junctions in our data. Newly-identified splice sites show little evidence of evolutionary conservation, suggesting that the majority are due to erroneous splice site choice. We show that sequence motifs involved in the recognition of exons are enriched in the vicinity of unconserved splice sites. We estimate that the average intron has a splicing error rate of approximately 0.7% and show that introns in highly expressed genes are spliced more accurately, likely due to their shorter length. These results implicate noisy splicing as an important property of genome evolution.


No excess of mitochondrial DNA deletions within muscle in progressive multiple sclerosis.
Mitochondrial dysfunction is an established feature of multiple sclerosis (MS). We recently described high levels of mitochondrial DNA (mtDNA) deletions within respiratory enzyme-deficient (lacking mitochondrial respiratory chain complex IV with intact complex II) neurons and choroid plexus epithelial cells in progressive MS. The objective of this paper is to determine whether respiratory enzyme deficiency and mtDNA deletions in MS were in excess of age-related changes within muscle, which, like neurons, are post-mitotic cells that frequently harbour mtDNA deletions with ageing and in disease. In progressive MS cases (n=17), known to harbour an excess of mtDNA deletions in the central nervous system (CNS), and controls (n=15), we studied muscle (paraspinal) and explored mitochondria in single fibres. Histochemistry, immunohistochemistry, laser microdissection, real-time polymerase chain reaction (PCR), long-range PCR and sequencing were used to resolve the single muscle fibres. The percentage of respiratory enzyme-deficient muscle fibres, mtDNA deletion level and percentage of muscle fibres harbouring high levels of mtDNA deletions were not significantly different in MS compared with controls. Our findings do not provide support to the existence of a diffuse mitochondrial abnormality involving multiple systems in MS. Understanding the cause(s) of the CNS mitochondrial dysfunction in progressive MS remains a research priority.


A retrospective analysis of cross-reacting cetuximab IgE antibody and its association with severe infusion reactions.
Severe infusion reactions (SIRs) at rates of 5% or less are known side effects of biological agents, including mAbs such as cetuximab. There are currently no prospectively validated risk factors to aid physicians in identifying patients who may be at risk of experiencing an SIR following administration of any of these drugs. A retrospective analysis of 545 banked serum or plasma samples from cancer patients participating in clinical trials of cetuximab was designed to evaluate whether the presence of pretreatment IgE antibodies against cetuximab, as determined by a commercially available assay system, is associated with SIRs during the initial cetuximab infusion. Patients with a positive test indicating the presence of pretreatment antibodies had a higher risk of experiencing an SIR; however, at the prespecified cutoff utilized in this analysis, the test has a relatively low-positive predictive value (0.577 [0.369-0.766]) and a negative predictive value of 0.961 (0.912-0.987) in an unselected patient population. Data collected in this large retrospective validation study support prior observations of an association between the presence of pretreatment IgE antibodies cross-reactive with cetuximab and SIRs. Further analysis of the test's ability to predict patients at risk of an SIR would be required before this assay could be used reliably in this patient population.


Research notes: Effect of environmental temperature on broiler chickens subjected to growth restriction at an early age.
An experiment was carried out to evaluate the effect of early growth restriction (EGR) induced by feed restriction on the ability of male broiler chickens to withstand exposure to high environmental temperatures. A 2 x 4 factorial arrangement of treatments was employed to determine whether such an exposure affects the beneficial effect of EGR on performance and carcass quality. Chicks consumed feed ad libitum or were feed-restricted from 6 to 12 d of age (to achieve 60% growth of the ad libitum birds). From 4 to 8 wk of age, the chickens were kept in temperature-controlled chambers at 25, 30, 35 C and a diurnal cyclic temperature of 25:35 C. Weight gain, feed intake, and feed efficiency from 6 to 8 wk of age (after acclimatization to the various temperatures) and BW at 8 wk of age were significantly (P < or = 0.05) depressed by the high temperatures (35 and 25:35 C) compared with values observed at 25 and 30 C. Early growth restriction did not affect these variables. Relative heart, liver, and breast meat weights at 8 wk of age were decreased significantly with increasing temperature. Relative liver weight was also decreased significantly in the EGR chicks, whereas breast yield was increased significantly in these birds. Relative abdominal fat pad size was not affected by increasing the temperature from 25 to 35 C, but it was significantly lower in chickens kept at 25:35 C than in those kept at 25 and 35 C. Early growth restriction significantly reduced the size of abdominal fat. Because no significant interactions between EGR and ambient temperature on the above mentioned variables were observed during 4 to 8 wk of age, it is concluded that EGR can not alleviate the adverse effect of high ambient temperature on performance, nor can high ambient temperature obliterate the beneficial effect of EGR in improving feed efficiency and reducing fattening.


Pulse wave travel distance as a novel marker of ventricular-arterial coupling.
Each stroke volume ejected by the heart is distributed along the arterial system as a pressure waveform. How far the front of the pressure waveform travels within the arterial system depends both on the pulse wave velocity (PWV) and the ejection time (ET). We tested the hypothesis that ET and PWV are coupled together, in order to produce a pulse wave travel distance (PWTD = PWV × ET) which would match the distance from the heart to the most distant site in the arterial system. The study was conducted in 11 healthy volunteers. We recorded lead II of the ECG along with pulse plethysmography at ear, finger and toe. The ET at the ear and pulse arrival time to each peripheral site were extracted. We then calculated PWV followed by PWTD for each location. Taken into account the individual subject variability PWTDToe in the supine position was 153 cm (95% CI 146-160 cm). It was not different from arterial pathway distance from the heart to the toe (D Toe 153 cm). The PWTDFinger and PWTDEar were longer than the distance from the heart to the finger and ear irrespective of body position. ETEar and PWVToe appear to be coupled in healthy subjects to produce a PWTD that is roughly equivalent to the arterial pathway distance to the toe. We propose that PWTD should be evaluated further to test its potential as a noninvasive parameter of ventricular-arterial coupling in subjects with cardiovascular diseases.