The impact of perceived treatment assignment on smoking cessation outcomes among African-American smokers.
The double-blind placebo-controlled design is commonly considered the gold standard in research methodology; however, subject expectation bias could subvert blinding. The primary aim of this study was to examine the impact of expectation bias. Specifically, we examined perceived treatment assignment on smoking cessation outcome rates among participants enrolled in a clinical trial of bupropion (150 mg SR, BID). Analyses were conducted on data collected during "Kick It at Swope," a double-blind, placebo-controlled, randomized trial of 600 African-American smokers. Chi-square and multiple logistic regression analyses were used to examine the impact of perception of assignment on treatment effect and cotinine-verified smoking abstinence rates. Participants were predominantly middle-aged (mean 44.7, SD 11.2), African-American women (68.6%), who smoked 19 CPD (SD = 8.1). Most had completed at least a high school education or GED (51.6%), and 55% had a monthly family income <$1,800. At week 6 (end of treatment) and week 26 (end of study), participants were asked to report their perceived treatment group assignment. Self-reported abstinence (weeks 6 and 26) was confirmed using CO and cotinine biochemical verification. After adjusting for actual treatment assignment, age and baseline cotinine, participants who perceived being assigned to bupropion vs. placebo were more likely to be abstinent at weeks 6 (OR = 2.07, 95% CI: 1.29 to 3.33, p = 0.002) and 26 (OR = 1.85, 95% CI: 1.05 to 3.24, p = 0.032). Results support previous research that expectation bias associated with judgment of treatment assignment is a strong predictor of outcome and confirms this relationship in a smoking cessation trial using bupropion SR among African-American smokers.


Changes in the Th1:Th2 cytokine bias in pregnancy and the effects of the anti-inflammatory cyclopentenone prostaglandin 15-deoxy-Δ(12,14)-prostaglandin J2.
Pregnancy is a complex immunological state in which a bias towards T helper 2 (Th2) protects the fetus. Evidence suggests that proinflammatory cytokines increase the risk of poor neonatal outcome, independently of the direct effect of preterm labour. The anti-inflammatory prostaglandin 15-deoxy-Δ(12,14)-Prostaglandin J(2) (15dPGJ(2)) inhibits nuclear factor Kappa B (NF-κB) in amniocytes and myocytes in vitro and is a ligand for the chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) receptor. Here we examine the Th1:Th2 cytokine bias in pregnancy and whether 15dPGJ(2) could be used to inhibit the production of the proinflammatory cytokines through inhibition of NF-κB while simultaneously promoting Th2 interleukin 4 (IL-4) synthesis via CRTH2 in T helper cells. Peripheral blood mononuclear cells (PBMCs) from women at 28 weeks, term pre-labour, term labour as well as non-pregnant female controls were cultured with 15dPGJ(2) or vehicle control and stimulated with phorbol myristyl acetate (PMA)/ionomycin. The percentage of CD4(+) cells producing interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) in response to PMA/ionomycin was significantly reduced in pregnancy. 15dPGJ(2) reduced IFN-γ and TNF-α production in stimulated T helper cells, but did not alter IL-4 production in CRTH2(+ve) cells. 15dPGJ(2) also reduced phospho-p65 in stimulated PBMCs. In summary, 15dPGJ(2) suppresses the Th1 response of PBMCs during pregnancy and active labour whilst maintaining the Th2 response suggesting a therapeutic benefit in reducing neonatal morbidity in inflammation-induced PTL.


[Microsurgical fascicular neurorrhaphy. Our method].
The author holds the microsurgical fascicular neurorrhaphy superior to the classical epineural suture. Microsurgical technique is more time-consuming and requires greater experience, moreover the application of autologous nerve grafts is often necessary. In secondary reconstructive operations of peripheral nerves a strong epineurium was found and in cases of small defects a surgical procedure was devised which permitted immediate microsurgical nerve suture without autologous nerve grafts. By isolating the epineurium it was possible to suture it initially on one side of the nerve, giving relaxation of the fascicle groups for direct repair.


Random matrix approach to collective behavior and bulk universality in protein dynamics.
Covariance matrices of amino acid displacements, commonly used to characterize the large-scale movements of proteins, are investigated through the prism of random matrix theory. Bulk universality is detected in the local spacing statistics of noise-dressed eigenmodes, which is well described by a Brody distribution with parameter beta approximately = 0.8. This finding, supported by other consistent indicators, implies a novel quantitative criterion to single out the collective degrees of freedom of the protein from the majority of high-energy, localized vibrations.


Development in children following abuse and nonorganic failure to thrive.
Thirty-nine children who had been abused an average of 5 1/2 years earlier and 14 children who had been admitted to the hospital with nonorganic failure to thrive (NOFTT) 13 years earlier were studied to look at similarities and differences in their development. Each group was studied in relation to a comparison group matched for age, sex, social class, and ethnic background. The abused children and those with NOFTT were similar in their language ability, and were significantly behind their comparison groups in language development, reading age, and verbal intelligence. The abused children, but not the group who had NOFTT, were significantly behind their comparison group in general intelligence, interpersonal relations, and self-concept, but in contrast to the children with NOFTT they were not delayed in social maturity. The long-term adverse sequelae of these two conditions emphasize the need for a long-term, child-centered approach.


Effect of cysteine on oltipraz blood levels in green monkeys (Cercopithecus aethiops).
Oral coadministration of oltipraz, an antischistosomal compound, with cysteine to green monkeys (Cercopithecus aethiops) led to a marked increase in both the extent and rate of oltipraz bioavailability. The drug blood levels were monitored using a single extraction gas-liquid chromatographic assay. When 6 healthy adult animals were given oltipraz together with cysteine in a crossover study, peak serum concentrations, areas under the curve and absorption rate constants of oltipraz were on average 7 times greater than when the drug was administered alone. Oltipraz peak serum concentrations were reached 1 h earlier as a result of cysteine administration (2 h after dosing). At present, the mechanisms responsible for the effect(s) of cysteine on oltipraz bioavailability have not been identified. The marked increase in oltipraz bioavailability produced by coadministration of cysteine, irrespective of the exact mechanisms involved, may have significant clinical implications with regard to the treatment of schistosomiasis. Our results also indicate that oltipraz blood levels seem to be influenced by some sex-related factors. Male monkeys had higher oltipraz blood levels than females. These sex-induced differences were more evident in oltipraz-cysteine-treated monkeys.


Fast multispectral diffuse optical tomography system for in vivo three-dimensional imaging of seizure dynamics.
We describe a multispectral continuous-wave diffuse optical tomography (DOT) system that can be used for in vivo three-dimensional (3-D) imaging of seizure dynamics. Fast 3-D data acquisition is realized through a time multiplexing approach based on a parallel lighting configuration--our system can achieve 0.12 ms per source per wavelength and up to a 14 Hz sampling rate for a full set of data for 3-D DOT image reconstruction. The system is validated using both static and dynamic tissue-like phantoms. An initial in vivo experiment using a rat model of seizure is also demonstrated.


[Influence of different procedures of alimentary tract reconstruction after total gastrectomy for gastric cancer on the nutrition and metabolism of patients: a prospective clinical study].
To investigate the influence of different alimentary tract reconstruction procedures after total gastrectomy for treatment of gastric cancer on nutrition and metabolism and explore an ideal reconstruction procedure. A total of 149 patients with gastric cancer who had undergone total gastrectomy were randomly allocated into 5 groups of 30 patients (except the group HLD with 29 cases) to undergo 5 different alimentary tract reconstruction procedures: simple esophagojejunostomy using Roux-en-Y technique (RY), P pouch with Roux-en-Y reconstruction (PRY), jejunal pouch reconstruction according to Hunt-Lawrence technique (HL), jejunal pouch original interposition reconstruction modified by the authors (JOP), and Hunt-Lawrence reconstruction technique maintaining duodenal passage (HLD). Three and six months after operation, quality of life (Visick grade), PNI; body weight; and serum nutritional parameters, including albumin, (ALB), total protein (TP), transferrin (TF), hemoglobin (HB), and serum iron (SI), were evaluated. In comparison with those of the PRY, HL, JOP, and HLD groups, the patients of the RY group show greater weigh loss, and lower ALB, TP, and TF (all P < 0.05). The HB, SI, and TS levels in the JOP group and HLD group were significantly higher than those in the RY, PRY, and HL groups (all P < 0.05). Different procedures of alimentary tract reconstruction after total gastrectomy have great influence on the patients' nutrition at different degrees. The patients undergoing the procedures with a reservoir show higher serum nutritional parameters and better body weight. The volume of reservoir has no major clinical importance. The jejunal pouch original interposition reconstruction modified by the authors (JOP), constructing a gastric reservoir and maintaining the alimentary tract flowing through the duodenum is an ideal choice for the reconstruction after total gastrectomy.


Cloning and characterization of the immunity region of phage phi 80.
The immunity region of phage phi 80 has been localized. It codes for at least three proteins: a protein of 34 kDa which has the biological properties of the phage repressor, and two other proteins of 9 kDa and 18 kDa which are the first proteins on the rightward operon. These two proteins are negatively regulated by the 34 kDa protein at a divergent promoter site. By position analogy with phage lambda, but not by its biological activity, the 9 kDa protein could be the cro product. The 18 kDa protein is able to block totally UV induction of phage phi 80.


MRI findings in methanol intoxication: a report of three cases.
Methanol is a highly toxic substance and acute methanol poisoning produces severe metabolic acidosis and serious neurological symptoms, including severe visual impairment, extrapyramidal signs and coma. We present three cases of accidental methanol intoxication and discuss the MRI findings.


Effectiveness and safety of mepolizumab in combination with corticosteroids in patients with eosinophilic granulomatosis with polyangiitis.
Mepolizumab (MPZ), an anti-interleukin-5 antibody, is effective for the treatment of eosinophilic granulomatosis with polyangiitis (EGPA). However, its effectiveness has not been adequately evaluated in real-world clinical practice. In this study, we assessed the effectiveness and safety of MPZ (300 mg) for relapsing/refractory EGPA resistant to corticosteroids (CS) for 1 year in real-world settings. We administered MPZ (300 mg) to 16 patients with relapsing/refractory EGPA resistant to CS (Post-MPZ). We also retrospectively collected data from the same patients for the 12 months before the administration of MPZ (Pre-MPZ). The primary endpoint was the 12-month remission rate after MPZ administration and the secondary endpoints were the Birmingham vasculitis activity score (BVAS), vasculitis damage index (VDI), eosinophil counts, changes in concomitant CS doses/concomitant immunosuppressant use, MPZ retention rate, and incidence of adverse events. The clinical course was compared between Pre-MPZ and Post-MPZ. The 12-month remission rate after the initiation of MPZ was 75%. No change was observed in BVAS, eosinophil count, or concomitant CS dose over time in the Pre-MPZ group, whereas all these parameters were significantly decreased over time in the Post-MPZ group. The number of patients using concomitant immunosuppressant also decreased over time in the Post-MPZ group. VDI did not increase in either group. The MPZ retention rate was 100% and only three patients (18.8%) had infections. Changes in BVAS, eosinophil count, and cumulative concomitant CS dose were significantly lower in the Post-MPZ group than in the Pre-MPZ group. There was no significant difference in the changes in VDI between the groups. This study demonstrated that MPZ is effective and safe for EGPA. Furthermore, MPZ decreases disease activity, increases remission rate, and has a CS-sparing effect.


A suspension technique for efficient large-scale cancer organoid culturing and perturbation screens.
Organoid cell culture methodologies are enabling the generation of cell models from healthy and diseased tissue. Patient-derived cancer organoids that recapitulate the genetic and histopathological diversity of patient tumours are being systematically generated, providing an opportunity to investigate new cancer biology and therapeutic approaches. The use of organoid cultures for many applications, including genetic and chemical perturbation screens, is limited due to the technical demands and cost associated with their handling and propagation. Here we report and benchmark a suspension culture technique for cancer organoids which allows for the expansion of models to tens of millions of cells with increased efficiency in comparison to standard organoid culturing protocols. Using whole-genome DNA and RNA sequencing analyses, as well as medium-throughput drug sensitivity testing and genome-wide CRISPR-Cas9 screening, we demonstrate that cancer organoids grown as a suspension culture are genetically and phenotypically similar to their counterparts grown in standard conditions. This culture technique simplifies organoid cell culture and extends the range of organoid applications, including for routine use in large-scale perturbation screens.


N-trifluoroacetyl sialyl phosphite donors for the synthesis of alpha(2 --> 9) oligosialic acids.
[reaction: see text] A new method for the synthesis of alpha(2 --> 9) oligosialic acids is developed using phosphite sialyl donors that are protected with a C-5 N-trifluoroacetyl (NHTFA) substituent. Compared with conventional donors, these donors gave a higher degree of alpha-anomeric selectivity during glycosidic bond formation and better yields during iterative sialylation in the synthesis of oligosialic acids.


ADHD among a cohort of ethnic minority women.
Attention deficit-hyperactivity disorder has focused primarily on Caucasian boys and men and resulted in limited insight about the experiences of attention deficit-hyperactivity disorder among women from an ethnically diverse background. This descriptive qualitative study explored the experience of 16 women diagnosed with attention deficit-hyperactivity disorder who were also engaged in their academic pursuits. Three themes emerged: (i) internalized chaos, (ii) cultivation of self-understanding, and (iii) commitment to building capacity. Comorbid psychological disorders (e.g., major depression and anxiety) among women with attention deficit-hyperactivity disorder were associated with increased impairment. Broader criteria that demonstrate equitable assessment and recognition of attention deficit-hyperactivity disorder symptoms for both genders are requisite to promote effective interventions and recovery.


Integration of gene maps: chromosome 21.
Physical, cytogenetic, and genetic data including microsatellite markers and a covering sequence-tagged site (STS) map have been entered into a location database and integrated into a summary map that subsumes a composite physical location, sex-specific genetic locations, cytogenetic and regional assignments, mouse homology, rank, and references. With the omission of 52 loci whose location is known only from cytogenetic assignment to an interval greater than 10 megabases, there are 198 loci in the covering STS map and an additional 145 loci. The physical length is consistent with 11 megabases for 21p and 39 megabases for 21q. With error filtration and allowance for high interference, the genetic length in males corresponds to the chiasma map (54.7 centimorgans), and the genetic length in females is 76.5 centimorgans. The relation between map integration and the STS paradigm is illustrated and discussed.


Congenital dyserythropoietic anemia type II. Radioautographic, ultrastructural and cytogenetical study in two familial cases.
The case of two sisters is reported in whom morphological (multinucleated erythroblasts in the bone marrow) and serological (positive Ham test, negative sucrose and water-sugar tests) characteristics of a congenital dyserythropoietic anemia type II were found. Radioautographic, cytogenetic and electron microscope studies confirmed the presence of an inefficient erythropoiesis and of a membrane defect, common in this disease. The finding of a nil LAP score has never been reported so far.


A universal matrix-assisted laser desorption/ionization cleavable cross-linker for protein structure analysis.
The concept of protein cross-linking in combination with mass spectrometry holds great promise to derive structural information on protein conformation and protein-protein interactions. We recently presented a dissociative amine-reactive cross-linker (NHS-BuUrBu-NHS) that is shown herein to be universally applicable to protein structure analysis under matrix-assisted laser desorption/ionization tandem mass spectrometric (MALDI-MS/MS) conditions, based on the examples of the peptides substance P, luteinizing hormone releasing hormone (LHRH), and the 32-kDa ligand-binding domain of peroxisome proliferator-activated receptor alpha (PPARα). The characteristic fragment ion patterns and constant neutral losses of the cross-linker greatly simplify the identification of different cross-linked species from complex mixtures and drastically reduce the potential of identifying false-positive cross-links. Therefore, this cross-linker holds an enormous potential for deriving structural information of proteins and protein complexes in a highly automated fashion.


Two cases of penile necrosis in diabetics with end-stage renal failure.
Distal penile necrosis associated with renal failure is a rare entity; only a few cases have been reported in the literature. Penile necrosis can frequently be a difficult management problem, the etiology of which is infectious, traumatic, or vascular. Physiological abnormalities are usually found in association with this condition, including diabetes, hyperparathyroidism, and peripheral vascular disease. Penile necrosis is a poor prognostic factor associated with high morbidity. We report two cases of this condition, presenting the clinical and pathophysiological background.


The presence of sinapyl lignin in Ginkgo biloba cell cultures changes our views of the evolution of lignin biosynthesis.
Suspension cell cultures (SCCs) from one of the oldest seed plants, Ginkgo biloba, show unpredictable alterations in the nature of the lignins, such as is the recruitment of sinapyl alcohol for lignin biosynthesis, compared with the woody tissues of the same species, which lack syringyl (S) lignins. These results show that, in this gymnosperm, the genes involved in sinapyl alcohol biosynthesis are latent and that their regulatory regions respond, by initiating gene expression, to the developmental signals and the environmental clues, which condition its in vitro culture. G. biloba SCCs not only synthesize S lignins but also their extracellular proteome contains both class III peroxidases capable of oxidizing sinapyl alcohol and enzymes involved in H2O2 production, observation which suggests that the peroxidase branch for the oxidative coupling of sinapyl alcohol units into lignins is operative. The incomplete knowledge of the G. biloba peroxidase-encoding genes led us to purify, characterize and partially sequence the peroxidase responsible for monolignol oxidation. When the major peroxidase from G. biloba SCCs (GbPrx) was purified to homogeneity, it showed absorption maxima in the visible region at 414 (Soret band), and at 543 and 570 nm, which calls to mind those shown by low-spin ferric peroxidases. However, the results also showed that the paraperoxidase-like character of GbPrx is not an obstacle for oxidizing the three monolignols compared with high-spin ferric peroxidases. Taken together, these results mean that the time at which the evolutionary gain of the segment of the route that leads to the biosynthesis of S lignins took place in seed plants needs to be revised.


Turbulent boundary layer under the control of different schemes.
This work explores experimentally the control of a turbulent boundary layer over a flat plate based on wall perturbation generated by piezo-ceramic actuators. Different schemes are investigated, including the feed-forward, the feedback, and the combined feed-forward and feedback strategies, with a view to suppressing the near-wall high-speed events and hence reducing skin friction drag. While the strategies may achieve a local maximum drag reduction slightly less than their counterpart of the open-loop control, the corresponding duty cycles are substantially reduced when compared with that of the open-loop control. The results suggest a good potential to cut down the input energy under these control strategies. The fluctuating velocity, spectra, Taylor microscale and mean energy dissipation are measured across the boundary layer with and without control and, based on the measurements, the flow mechanism behind the control is proposed.


A semiautomatic tool for 3D medical image analysis using active contour models.
We describe a computerized tool designed to improve the efficiency with which clinicians can delineate volumes of interest in multi slice medical data such as NMR and CT images. The tool allows a trained user to create deformable models that can be used to search for edges or regions. Models can also be copied from previously analysed images into fresh slices, and frozen so they can only translate, rotate and scale, without deforming. In this way an accurate model can be used to extract an object from several adjacent slices. Surfaces are then reconstructed from the models using conic splines. By applying these techniques to objects such as tumours, it should be possible to generate three-dimensional images that are accurate enough to use when planning surgery and radiotherapy.


Intradural clival chordoma: a case report.
Clival chordoma is a rare intracranial neoplasm located in the clivus with bony extension and destruction. It is difficult to resect completely and generally has a poor prognosis. However, intradural clival chordomas have been reported with good surgical outcomes. We present a rare case of intradural chordoma and a review of the literature.


Multiple endocrine neoplasia 1: a broad overview.
This review article discusses the diagnoses and treatment of patients with multiple endocrine neoplasia type 1 (MEN 1). The most common tumors associated with MEN 1 are located in the pancreas, pituitary, and parathyroid glands. Less common tumors include neuroendocrine tumors of the lung and thymus, adrenal tumors, and cutaneous lesions. This article describes the diagnosis, clinical manifestations, treatment, and surveillance of tumors associated with patients who are diagnosed with MEN 1.


New Lanostane-Type Triterpenes with Anti-Inflammatory Activity from the Epidermis of Wolfiporia cocos.
A chemical study on the epidermis of cultivated edible mushroom Wolfiporia cocos resulted in the isolation and identification of 46 lanostane triterpenoids, containing 17 new compounds (1-17). An experimental determination of their anti-inflammatory activity showed that poricoic acid GM (39) most strongly inhibited NO production in LPS-induced RAW264.7 murine macrophages with an IC50 value at 9.73 μM. Furthermore, poricoic acid GM induced HO-1 protein expression and inhibited iNOS and COX2 protein expression as well as the release of PGE2, IL-1β, IL-6, TNF-α, and reactive oxygen species (ROS) in LPS-induced RAW264.7 cells. Mechanistically, poricoic acid GM suppressed the phosphorylation of the IκBα protein, which prevented NF-κB from entering the nucleus to lose transcriptional activity and inhibited the dissociation of Keap1 from Nrf2, thereby activating Nrf2 into the nucleus to regulate antioxidant genes. Furthermore, the MAPK signaling pathway may play a significant role in poricoic acid GM-induced elimination of inflammation. This work further confirms that lanostane triterpenoids are key ingredients responsible for the anti-inflammatory properties of the edible medicinal mushroom W. cocos.


Relationship between birthweight and blood lipid concentrations in later life: evidence from the existing literature.
It has been suggested that there is a link between fetal growth and chronic diseases later in life. Several studies have shown a negative association between birthweight and cardiovascular diseases, as well as cardiovascular disease risk factors, such as blood pressure and type 2 diabetes. Far fewer studies have focused on the association between size at birth and blood lipid concentrations. We have conducted a qualitative assessment of the direction and consistency of the relationship between size at birth and blood lipid concentrations to see whether the suggested relationship between intrauterine growth and cardiovascular diseases is mediated by lipid metabolism. A literature search covering the period January 1966 to January 2003 was performed using Medline, Embase, and Web of Science. All papers written in English and reporting the relationship between size at birth and lipid levels in humans were assessed. Bibliographies were searched for further publications. From an initial screen of 1198 references, 39 papers were included involving 28 578 individuals. There was no consistent relationship between size at birth and blood lipid levels; the one exception being triglyceride concentration, which showed statistically significant negative or U-shaped, but not positive, relationships with birthweight. This review does not strongly support a link between birthweight and blood lipid levels in later life. However, the research in this area is limited and in order to make any definitive conclusions, longitudinal studies with sufficient power, data, and prospective follow-up are needed.


[Acetylcholinesterase in the erythrocyte membrane].
Acetylcholinesterase (AchE) in erythrocytes is one of the typical extraneural AchEs. Its activity is high in human erythrocytes. It has been known that the enzyme is present only in the membrane in erythrocytes and that it is localized on the outer side of the membrane. Recent studies have disclosed much of the primary structure of AchE and the membrane anchor structure. However, the physiological functions of erythrocyte AchE are still totally unknown. Nevertheless, the enzyme may be regarded as a model of AchE in the nervous system. In addition, it may be used to examine the status of the erythrocyte membrane. The enzyme activity seems useful as an indicator of the effects of AchE inhibitors, such as pesticides. It should be noted, however, that the activity of AchE in erythrocytes is not always a good indicator of intoxication with AchE inhibitors.


Alterations in protein synthetic capability of nerve cells in Alzheimer's disease.
Cytoplasmic RNA content, nuclear and nucleolar volume are all significantly reduced in nerve cells of the temporal cortex in cases of Alzheimer's disease, examined both at diagnostic craniotomy and post mortem, when compared with non=demented control cases of similar age. On average, at necropsy, all three parameters are equally reduced by about 40-50%, but in biopsy cases, nuclear volume is decreased by the greatest amount (43%), followed by nucleolar volume is decreased by the greatest amount (43%), followed by nucleolar volume (36%), and cytoplasmic RNA (26%). These findings indicate that a change in protein synthesis occurs daily in the course of Alzheimer's disease which may result from a primary alteration within the nuclear apparatus.


Natural Poly- and Oligosaccharides as Novel Delivery Systems for Plant Protection Compounds.
To increase the bioavailability of plant protection products, we have applied a new approach based on noncovalent association with natural water-soluble polysaccharides and oligosaccharides as delivery systems (DSs). The mechanochemical technique has been applied to prepare the solid-state nanodispersed compositions of antidote 1,8-naphthalic anhydride (NA) with arabinogalactan, sodium salt of carboxymethylcellulose, and glycyrrhizin as DSs. The effect of DSs on the solubility and the penetration of NA into the seeds of barley and wheat has been investigated by various physicochemical techniques. All DSs considerably enhance the solubility of NA and improve its penetration into the grain. The influence of polysaccharides and oligosaccharides on artificial lipid membranes was studied by the NMR relaxation method. It was concluded that the effect of polysaccharides and oligosaccharides on the penetration efficacy of plant protection products might be associated with the detected solubility enhancement and the affinity of DSs to the surface of cell membranes.


Risk factors involved in orofacial cleft predisposition - review.
Clefts that occur in children are a special topic. Avoiding risk factors, and also an early diagnosis of cleft possibility can result in minimizing or avoiding them. If on the other hand when clefts occur they require a long-term, multistage specialized treatment. Etiology of clefts seems to be related to many factors. Factors such as genetic, environmental, geographic and even race factors are important. Identification of risk factors can lead to prevention and prophylactic behaviors in order to minimize its occurrence. Exposure to environmental factors at home and work that lead to cleft predisposition should not be disregarded. It seems that before planning a family it would be wise to consult with doctors of different specializations, especially in high-risk families with cleft history in order to analyze previous lifestyle. Clefts are very common in hereditary facial malformations and are causing a lot of other irregularities in the head and neck region. In this paper after a brief papers review authors present socio-geographic, environmental and also work place related factors that are influencing pregnant women condition and should be taken under serious consideration.


Subgroups of smokers with different success rates after use of transdermal nicotine.
To identify subgroups of smokers with different success rates, we applied "tree-structured survival analysis" (TSSA) to data from a previously published trial of transdermal nicotine. The subjects who received active treatment (14 mg patch, n = 275, or the 21 mg patch, n = 262) constituted the sample for this analysis. Using age, gender, the Fagerstrom Tolerance Questionnaire (FTQ), motivation to quit, number of cigarettes smoked at baseline, and body mass index (BMI) as classification variables, TSSA identified two subgroups within the 14 mg patch group and four subgroups of smokers within the 21 mg patch group. Among those receiving the 14 mg patch, individuals with a BMI greater than 26.4 kg/m2 relapsed sooner than did those with a BMI less than or equal to this value. Within the 21 mg patch group, the survival curve for males was significantly different from that observed in females, with males experiencing a longer time to relapse after treatment than did females. Among females, those with a higher motivation to quit relapsed more slowly than did those women with less motivation to quit. This information may be helpful to clinicians seeking to match specific patients to specific treatments with traditional nicotine in order to maximize treatment outcomes.


Immobilized heparinase: In vitro reactor model.
Heparinase immobilized to agarose has previously been shown to be useful in degrading heparin and thereby preventing thromboembolytic complications when this anticoagulant has been used in extracorporeal perfusions. The current study examined the kinetics of this immobilized enzyme. When heparinase is covalently bound to 8% agarose, the partition coefficient of heparin in the catalytic particle is 0.36 +/- 0.048 (N = 10). The immobilized enzyme has a K(m) of 0.15 +/- 0.03 mg/mL and an activation energy of 10.3 +/- 0.57 kcal/gmol (N = 5). These values are statistically indistinguishable from the values for the free enzyme. The immobilized enzyme showed a pH activity optimum between 7.0 and 7.4, compared to the optimum pH of 6.5 for the soluble enzyme. The activity optimum of immobilized heparinase with respect to salt concentration was between 0 and 0.1M. A reactor containing immobilized heparinase recirculating internally at 1300 mL/min behaved as a continuously stirred tank reactor (CSTR) when solutions at a flow rate of 120 mL/min were passed through the device. The residence time distribution was determined using blue dextran (molecular weight 2 x 10(6) daltons), which is sterically excluded from the agarose catalyst. A model of the heparinase reactor based on ideal CSTR behavior and the immobilized enzyme kinetic parameters was developed. It accurately predicted experimental conversions over a range of catalyst volumes, enzyme loadings, and substrate concentrations to within 7% in most cases and with a maximum deviation of 13%.


Volumetrical measurements in vestibular schwannoma, the influence of slice thickness and patient's repositioning.
Conservative treatment in vestibular schwannomas is mainly dependent on optimal tumor size determination. The first objective of this study was to establish interobserver and intraobserver variability and the accuracy and reproducibility of three different measurement methods: one bidimensional and two volumetrical. The second objective was to evaluate the influence of the use of different magnetic resonance imaging (MRI) slice thickness and the influence of patient's repositioning on the measurements' outcome. Two consecutive studies have been prospectively performed, both mainly concerning volumetrical measurements. Both studies were performed in a tertiary academic, multidisciplinary center. In the first study, 19 patients were included between March 1996 and May 2002, with a total of 52 scans. The second study comprised 14 patients. All patients in the first study had at least two MRI examinations performed according to a standard protocol (T1-weighted gadolinium-enhanced, slice thickness of 3 mm, and interslice gap of 0.3 mm). The population in the second study underwent a conservative wait and scan (W&S) treatment. Both studies are discussed separately. In the first study, all scans were measured by four investigators, two of whom performed the measurements twice using three different methods. The first method concerns a manually performed bidimensional surface measurement along the petrous pyramid. The second method concerns a semiautomatic volumetrical measurement on a computer, relying on contour detection, and the last method concerns a fully automatic volume reconstruction also performed on a computer using different gray shade scales. All 14 patients included in the second study underwent three magnetic examinations. Three different T1-weighted gadolinium-enhanced sequences were used: the first using a slice thickness of 1 mm, the second again with 1-mm slice thickness but after having repositioned the patient. In the third sequence, a slice thickness of 3 mm was used. All scans were measured by two investigators using the three different methods, as described previously. The manual surface method shows large intraobserver variability, and its reproducibility is significantly lower compared with volume measurements. Because of a relatively large systematic error in small tumors, sensitivity of growth detection is low. Both volumetrical methods are hardly interobserver- and intraobserver-dependent, and the gray shade method turned out to be the most accurate. Radiologic progression is only significant at a volume increase of at least 50%. The influence of patient repositioning is negligible, whereas the use of 1-mm slice thickness seems to be superior to a 3-mm slice thickness. The volumetrical gray shade method is the most accurate method to detect early tumor progression. As tumor increase of at least 50% is needed to be able to speak of statistically significant tumor growth, the absence of radiologic progression does not mean that there is no tumor growth. Repositioning of the patient has no influence on the measurements' outcome, whereas for optimal magnetic resonance imaging examinations, a 1-mm slice thickness protocol seems to be superior.


Indications for local excision of ampullary lesions associated with familial adenomatous polyposis.
The relative risk of periampullary carcinoma is high in patients with familial adenomatous polyposis (FAP). We examined the possibility of local treatment of the ampullary lesion of these patients in an early stage. From August 1991 to April 1993, 37 patients with FAP underwent endoscopic examination and biopsy of the ampulla. Local excision of the ampulla was performed in seven patients. Two of the seven patients were diagnosed with carcinoma in situ; the ages of the patients were 35 and 46 years. A 38-year-old woman had ampullary carcinoma infiltrating into the muscle of the sphincter of Oddi. These three cases showed a protruding change or deformity of the ampulla endoscopically. The laboratory data, cholangiogram, and abdominal ultrasonography were within normal limits. The superficial change of the ampulla along with the young age of the patients were considered to be factors that warranted further observation. Postoperative complications in two cases were treated successfully. Indications for local excision of the ampulla of patients with FAP would be protruding change of the ampulla, biopsy showing adenoma with severe atypia or carcinoma, in situ, no jaundice with or without subclinical abnormality of laboratory data, no apparent dilatation in the bile duct, and age of more than 35 years.


Dose responses from inhaled monodisperse aerosols of 244Cm2O3 in the lung, liver and skeleton of F344 rats and comparison with 239PuO2.
The purpose of this study was to obtain information on the alpha-particle dose-response relationship of 244Cm in rats. Rats were exposed briefly by inhalation to graded levels of monodisperse aerosols of 244Cm2O3 heat-treated at 1150 degrees C. The initial lung burden (ILB) of each animal was determined by the use of the gamma-ray-emitting radionuclide 243Cm in the aerosols. Seven groups of 84-day-old F344/Crl rats (a total of 637 males and 645 females) were exposed once to 244Cm2O3 or sham-exposed to filtered ambient air. Mean ILBs of all rats per group ranged from 0.51 +/- 0.17 (+/-SD) to 240 +/- 82 kBq kg-1 body weight. Mean lifetime alpha-particle doses to the lungs per group ranged from 0.20 +/- 0.069 (+/-SD) to 36 +/- 6.5 Gy. After death, each rat was radiographed and necropsied. Dose-related increases occurred in incidences of benign and malignant lung neoplasms, except for the groups of rats with higher mean ILBs that were examined histologically (98 +/- 18 and 240 +/- 77 kBq kg-1 body weight) in which survival was markedly decreased. Also, average alpha-particle doses of 0.0014 +/- 0.00058 (+/-SD) to 0.17 +/- 0.091 Gy and 0.18 +/- 0.007 to 1.6 +/- 1.1 Gy were also absorbed by the liver and skeleton, respectively, in the rats in the different exposure groups. Primary liver neoplasms occurred in several rats. However, the incidence of these lesions was not related to dose. Increased incidences of bone neoplasms occurred only in rats receiving higher doses to the skeleton. Excess numbers of rats with lung neoplasms per 10(4) Gy to the lung per group ranged from 760 +/- 430 (+/- SE) at a mean dose of 0.48 Gy to 84 +/- 16 at a mean dose of 37 Gy. Risk factors for the lowest and highest ILB kg-1 body weight groups were not considered reliable because of large errors associated with these calculations and the life-span shortening in the highest ILB kg-1 group. Inhaled 244Cm2O3 appeared to be about 50% less effective as a lung carcinogen in rats compared to 239PuO2 at similar doses.


An integrative bioinformatics approach reveals coding and non-coding gene variants associated with gene expression profiles and outcome in breast cancer molecular subtypes.
Sequence variations in coding and non-coding regions of the genome can affect gene expression and signalling pathways, which in turn may influence disease outcome. In this study, we integrated somatic mutations, gene expression and clinical data from 930 breast cancer patients included in the TCGA database. Genes associated with single mutations in molecular breast cancer subtypes were identified by the Mann-Whitney U-test and their prognostic value was evaluated by Kaplan-Meier and Cox regression analyses. Results were confirmed using gene expression profiles from the Metabric data set (n = 1988) and whole-genome sequencing data from the TCGA cohort (n = 117). The overall mutation rate in coding and non-coding regions were significantly higher in ER-negative/HER2-negative tumours (P = 2.8E-03 and P = 2.4E-07, respectively). Recurrent sequence variations were identified in non-coding regulatory regions of several cancer-associated genes, including NBPF1, PIK3CA and TP53. After multivariate regression analysis, gene signatures associated with three coding mutations (CDH1, MAP3K1 and TP53) and two non-coding variants (CRTC3 and STAG2) in cancer-related genes predicted prognosis in ER-positive/HER2-negative tumours. These findings demonstrate that sequence alterations influence gene expression and oncogenic pathways, possibly affecting the outcome of breast cancer patients. Our data provide potential opportunities to identify non-coding variations with functional and clinical relevance in breast cancer.


Significant Increase in Depression in Women With Primary Dysmenorrhea: A Systematic Review and Cumulative Analysis.
Women with primary dysmenorrhea are vulnerable to develop a depressive disorder, which is a common form of psycho-disturbance. However, clinical findings are inconsistent across studies, and the evidence has not been previously synthesized. This study aims to investigate whether primary dysmenorrhea is associated with a higher risk of depression via a cumulative analysis. Four electronic databases were systematically searched for the eligible studies. The combined effect was assessed by analyzing the relative risk (RR) and standard mean differences (SMD) with a 95% confidence interval (CI). This cumulative analysis was registered on the PROSPERO (ID: CRD42020169601). Of 972 publications, a total of 10 studies involving 4,691 participants were included. Pooled results from six included studies showed that primary dysmenorrhea was associated with a significant depressive disorder (RR = 1.72, 95%CI: 1.44 to 2.0, P < 0.001; heterogeneity: I 2 = 0%, P = 0.544). In addition, synthesis results from two studies provided the BDI scores suggested that dysmenorrhea had significantly higher scores when compared to non-dysmenorrhea (SMD = 0.47, 95% CI: 0.31-0.62, P < 0.001; heterogeneity: I 2 = 0%, P = 0.518). However, in the two studies providing the PROMIS T-Score, the pooled result showed that there was no significant difference between women with dysmenorrhea and those without dysmenorrhea (P = 0.466). The overall quality of the evidence in our study was judged to MODERATE. The present study has confirmed the positive relationship between primary dysmenorrhea and depression. Social supports and medical help from pain management physicians or psychologists are important interventions for women with dysmenorrhea-suffering depressive disorder.


Interpreting the results of a retrospective comparison of test and reference treatments in a randomized clinical trial setting.
The retrospective comparison of test and reference treatment arms in a randomized prospective clinical trial is potentially useful in economic modeling seeking to assess the cost effectiveness of alternative therapies. To enhance the credibility of such retrospective comparisons, we propose the application of the following adjustments to significance levels obtained from standard statistical methodology: (1) a significance test for the lower bound of the 95 % confidence interval for the observed difference, (2) a conservative Bonferroni method of adjustment for multiple comparisons, (3) an adjusted p-value calculated using Scheffe's single-step method, and (4) Bayesian 95 % credibility intervals with a prior centered at zero. These adjustments were applied to data from a randomized double-blind concurrent trial (SPD489-325) that established the efficacy and safety of lisdexamfetamine dimesylate (LDX) in children and adolescents with attention-deficit/hyperactivity disorder (ADHD). Prospectively planned analyses demonstrated that the reduction in the symptoms of ADHD was significantly greater than placebo in patients treated with either LDX or the reference treatment, osmotic-release oral system methylphenidate (OROS-MPH). Retrospective analyses showed that the improvement in the symptoms of ADHD was greater in patients treated with LDX than OROS-MPH. We now show that this observation remained significant after the application of the four statistical penalties. By adjusting the significance level, it is possible to compare quantitatively such retrospective results with prospectively defined comparisons. However, the qualitative level of such retrospective evidence should remain secondary to that obtained from prospectively specified comparisons in a randomized clinical trial.


The roles of JAK2/STAT3 signaling in fusion of the secondary palate.
Cleft palate has a multifactorial etiology. In palatal fusion, the contacting medial edge epithelium (MEE) forms the epithelial seam, which is subsequently removed with the reduction of p63. Failure in this process results in a cleft palate. We herein report the involvement of janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling in palatal fusion and that folic acid rescues the fusing defect by reactivating JAK2/STAT3. In closure of bilateral palatal shelves, STAT3 phosphorylation was activated at the fusing MEE and mesenchyme underlying the MEE. JAK2 inhibition by AG490 inhibited STAT3 phosphorylation and resulted in palatal fusion failure without removal of the epithelial seam, in which p63 and keratin 17 (K17) periderm markers were retained. Folic acid application restored STAT3 phosphorylation in AG490-treated palatal explants and rescued the fusion defect, in which the p63- and K17-positive epithelial seam were removed. The AG490-induced palatal defect was also rescued in p63 haploinsufficient explants. These findings suggest that JAK2/STAT3 signaling is involved in palatal fusion by suppressing p63 expression in MEE and that folate restores the fusion defect by reactivating JAK2/STAT3.


Cooperation among Numb, MDM2 and p53 in the development and progression of pancreatic cancer.
We study the expression of Numb, MDM2 and p53 for clinical significance in pancreatic cancer (PC) and their functional relationship in regulating biological behaviors of PC cells. IHC, IB and qRT-PCR were used to detect Numb, MDM2 and p53 expression in PC. Transfection and drug intervention were used to investigate their functional relationship in PC cells. IHC showed that Numb expression was negatively associated with tumor size, differentiation and UICC stage, while expression of MDM2 and p53 was positively associated with tumor T and UICC stages, respectively (P < 0.05). Numb was an independent prognostic indicator in PC (P < 0.05). Patients with Numb-positive expression or combined with MDM2-negative expression had a significantly better overall survival (P < 0.05). Altered expression of Numb can regulate wild-type but not mutant p53 expression, while MDM2 knockdown increased Numb but not mutant p53 protein level. Meanwhile, Numb knockdown increased chemoresistance but decreased activated p53 and cleaved-caspase-3 protein expression in gemcitabine-treated Capan-2 cells. Moreover, Numb co-immunoprecipitated with p53 to prevent p53 ubiquitin-dependent protein degradation and this ubiquitin-dependent regulation plays an important role in the coordinate function of these three proteins on cell invasion and migration in PC cells. Our study is the first to demonstrate the clinical significance and functional cooperation among Numb, MDM2 and p53 involved in the development and progression of PC.


Electronystagmography: a review of 17 years experience.
The clinical concepts of electronystagmography (ENG) and vestibular testing are simple. They are not mysterious, complicated, nor time consuming. Every patient afflicted with dizziness deserves such a recording as part of the vestibular workup. No one would consider the evaluation of a patient with a hearing loss complete without an audiogram, and in 1975 no patient with a complaint of dysequilibrium has been properly or sufficiently evaluated without an ENG recording of balance function.


Pure Shift Nuclear Magnetic Resonance: a New Tool for Plant Metabolomics.
Nuclear Magnetic Resonance (NMR) is one of the most powerful tools used in metabolomics. It stands as a highly accurate and reproducible method that not only provides quantitative data but also permits structural identification of the metabolites present in complex mixtures. Metabolic profiling by 1H NMR has proven useful in the study of various types of plant scenarios, which include the evaluation of crop conditions, harvest and post- harvest treatments, metabolic phenotyping, metabolic pathways, gene regulation, identification of biomarkers, chemotaxonomy, quality control, denomination of origin, among others. However, signal overlapping of the large number of resonances with expanded J-coupling multiplicities complicates the spectra analysis and its interpretation, and represents a limitation for classical 1H NMR profiling. In the last decade, novel NMR broadband homonuclear decoupling techniques through which multiplet signals collapse into single resonance lines - commonly called Pure Shift methods - have been developed to overcome the spectra resolution problem inherent to 1H NMR classical spectra. Here a step-by-step protocol of the plant extract preparation and the procedure to record optimal Pure Shift PSYCHE and SAPPHIRE-PSYCHE spectra in three different plant matrices - Vanilla plant leaves, potato tubers (S. tuberosum), and Cape gooseberries (P. peruviana) - is presented. The effect of the gain in resolution in metabolic identification, correlation analysis and multivariate analyses, as compared against classical spectra, is discussed.


Encephalomyocarditis virus 2C protein antagonizes interferon-β signaling pathway through interaction with MDA5.
Encephalomyocarditis virus (EMCV) is one of the most important picornavirus. It infects many mammalian species and causes encephalitis, myocarditis, neurologic diseases, diabetes and reproductive disorders in pigs. And it evolves mechanisms for escaping innate immune responses. But the viral pathogenesis has not been understood completely. In this study, we firstly found that EMCV protein 2C is a strong IFN-β antagonist that interacts with MDA5 to inhibit induction of the IFN-β signal pathway. The mutations in amino acid residue V26 of 2C decrease the inhibition of IFN-β promoter activity and lost the ability to interact with MDA5, compared with wild type 2C protein. The rescued viruses with mutations in 2C (rV26A and rK25-3A) induced significantly higher IFN-β mRNA and protein levels in PK-15, HEK-293A and N2a cells, compared to wild type EMCV and the repaired viruses rV26A(R) and rK25-3A(R). These data indicate that the amino acid residue V26 of EMCV 2C plays important roles in inhibiting type I IFN production by interacting with MDA5.


Contraceptive failure rates of etonogestrel subdermal implants in overweight and obese women.
To estimate the contraceptive failure rates of the etonogestrel subdermal contraceptive implant in overweight and obese women and compare failure rates with women of normal weight and women using intrauterine devices (IUDs). The Contraceptive CHOICE Project is a large prospective cohort study designed to promote the use of long-acting reversible contraceptive methods to reduce unintended pregnancies in the St Louis region. Participants are provided reversible contraception of their choice at no cost. We collected baseline height and weight of each participant. During each survey, participants were asked about missed menses and possible pregnancies. Any participant who suspected a pregnancy was asked to come in for urine pregnancy testing. Analysis includes the first 8,445 participants enrolled in CHOICE of which 1,168 chose the implant and 4,200 chose the IUD. Student's t test, χ test, and Kaplan-Meier survival curves were used to perform statistical analyses to estimate failure rates in overweight and obese women using the implant and IUDs. Of the women choosing the implant, 28% were overweight and 35% were obese. Of the women who chose an IUD, 27% were overweight and 35% were obese. The 3-year cumulative failure rates for implant and IUD users were less than one per 100 women-years and did not vary by body mass index. We found no decrease in the effectiveness of the implant in overweight or obese women. The implant may be offered as a first-line contraceptive method to any woman seeking a reversible and reliable birth control method.


Alterations in adolescent dopaminergic systems as a function of early mother-toddler attachment: A prospective longitudinal examination.
Early experiences have the potential for outsized influence on neural development across a wide number of domains. In humans, many of the most important such experiences take place in the context of the mother-child attachment relationship. Work from animal models has highlighted neural changes in dopaminergic systems as a function of early care experiences, but translational research in humans has been limited. Our goal was to fill this gap by examining the longitudinal associations between early attachment experiences (assessed at 2.5 years) and neural responses to risk and rewards during adolescence (assessed at 13 years). Adolescence is a developmental period where sensitivity to rewards has important implications for behavior and long-term outcomes, providing an important window to study potential influences of early attachment experiences on reward processing. In order to address this question, 50 adolescents completed a risk and reward task during an fMRI scan, allowing us to assess differences in neural sensitivity to changes in risk level and reward amount as a function of early attachment experiences. Adolescents with insecure attachment histories showed blunted sensitivity to increasing risk levels in regions of the dorsal striatum, while also showing heightened sensitivity to increasing reward levels in the same region. These results highlight the importance of early attachment experiences for long-term neural development. Specifically, early exposure to more maladaptive relationships with caregivers may confer dual risks prospectively for adolescents, sensitizing them to rewarding outcomes while de-sensitizing them to potential risks associated with those behaviors, perhaps due to stress-related dopaminergic changes early in development.


The Route of Ion and Water Movements Across the Gill Epithelium of the Freshwater Shrimp Macrobrachium olfersii (Decapoda, Palaemonidae): Evidence From Ultrastructural Changes Induced by Acclimation to Saline Media.
The ultrastructure of the pillar cells in the gill lamellae of the freshwater shrimp Macrobrachium olfersii was examined to evaluate the routes of salt and water movement across the gill epithelium and into the hemolymph. Alterations were morphometrically quantified in shrimp maintained in fresh water (FW, <0.5{permill} salinity) and after acclimation to saline media (21{permill} or 28{permill} salinity). The tissue interface between the hemolymph and the external medium consists exclusively of the thin apical flange regions of the pillar cells, the upper membrane of which is highly amplified by dense microvilli and overlain by a thin cuticle. The lower flange membrane, bathed by the hemolymph, is smooth and not invaginated. Contiguous flanges are strongly bound by junctional structures including desmosomes and septate junctions. The basal surface of the pillar cell perikaryon is linked to the adjacent septal cells through many basolateral junctions. The septal cell plasmalemma is abundantly and deeply invaginated, each infolding enclosing numerous mitochondria; these characteristics are typical of salt-transporting machinery. After shrimps were acclimated to saline media for 10 days, the thickness of the pillar cell flanges was significantly reduced (from 1.3 to {approx}0.4 {mu}m), as was the height (from 0.8 to 0.3 {mu}m) and density (from 4.0 to {approx} 1.8 microvilli/{mu}m) of the apical microvilli. This reduction in the apical surface area of the pillar cells appears to lead to decreased ionic permeability, concomitant with a reduction in Na+/K+-ATPase activity, thus limiting Na+ uptake. In contrast to the brachyurans, in which the respiratory and ion-transporting mechanisms are differentially located in the anterior and posterior gills, in palaemonid shrimps the pillar cells apparently play a dual role: ions move preferentially through ion transporters in the microvilli above the pillar cell perikaryon, while respiratory gases are exchanged through the fine flange regions in contact with the hemolymph.


hnRNP G/RBMX enhances HPV16 E2 mRNA splicing through a novel splicing enhancer and inhibits production of spliced E7 oncogene mRNAs.
Human papillomavirus type 16 (HPV16) E2 is an essential HPV16 protein. We have investigated how HPV16 E2 expression is regulated and have identifed a splicing enhancer that is required for production of HPV16 E2 mRNAs. This uridine-less splicing enhancer sequence (ACGAGGACGAGGACAAGGA) contains 84% adenosine and guanosine and 16% cytosine and consists of three 'AC(A/G)AGG'-repeats. Mutational inactivation of the splicing enhancer reduced splicing to E2-mRNA specific splice site SA2709 and resulted in increased levels of unspliced E1-encoding mRNAs. The splicing enhancer sequence interacted with cellular RNA binding protein hnRNP G that promoted splicing to SA2709 and enhanced E2 mRNA production. The splicing-enhancing function of hnRNP G mapped to amino acids 236-286 of hnRNP G that were also shown to interact with splicing factor U2AF65. The interactions between hnRNP G and HPV16 E2 mRNAs and U2AF65 increased in response to keratinocyte differentiation as well as by the induction of the DNA damage response (DDR). The DDR reduced sumoylation of hnRNP G and pharmacological inhibition of sumoylation enhanced HPV16 E2 mRNA splicing and interactions between hnRNP G and E2 mRNAs and U2AF65. Intriguingly, hnRNP G also promoted intron retention of the HPV16 E6 coding region thereby inhibiting production of spliced E7 oncogene mRNAs.


EORTC guidelines for the use of erythropoietic proteins in anaemic patients with cancer: 2006 update.
Anaemia is frequently diagnosed in patients with cancer, and may have a detrimental effect on quality of life (QoL). We previously conducted a systematic literature review (1996-2003) to produce evidence-based guidelines on the use of erythropoietic proteins in anaemic patients with cancer.[Bokemeyer C, Aapro MS, Courdi A, et al. EORTC guidelines for the use of erythropoietic proteins in anaemic patients with cancer. Eur J Cancer 2004;40:2201-2216.] We report here an update to these guidelines, including literature published through to November 2005. The results of this updated systematic literature review have enabled us to refine our guidelines based on the full body of data currently available. Level I evidence exists for a positive impact of erythropoietic proteins on haemoglobin (Hb) levels when administered to patients with chemotherapy-induced anaemia or anaemia of chronic disease, when used to prevent cancer anaemia, and in patients undergoing cancer surgery. The addition of further Level I studies confirms our recommendation that in cancer patients receiving chemotherapy and/or radiotherapy, treatment with erythropoietic proteins should be initiated at a Hb level of 9-11 g/dL based on anaemia-related symptoms rather than a fixed Hb concentration. Early intervention with erythropoietic proteins may be considered in asymptomatic anaemic patients with Hb levels 11.9 g/dL provided that individual factors like intensity and expected duration of chemotherapy are considered. Patients whose Hb level is below 9 g/dL should primarily be evaluated for need of transfusions potentially followed by the application of erythropoietic proteins. We do not recommend the prophylactic use of erythropoietic proteins to prevent anaemia in patients undergoing chemotherapy or radiotherapy who have normal Hb levels at the start of treatment, as the literature has not shown a benefit with this approach. The addition of further supporting studies confirms our recommendation that the target Hb concentration following treatment with erythropoietic proteins should be 12-13 g/dL. Once this level is achieved, maintenance doses should be titrated individually. There is Level I evidence that dosing of erythropoietic proteins less frequently than three times per week is efficacious when used to treat chemotherapy-induced anaemia or prevent cancer anaemia, with studies supporting the use of epoetin alfa and epoetin beta weekly and darbepoetin alfa given every week or every 3 weeks. We do not recommend the use of higher than standard initial doses of erythropoietic proteins with the aim of producing higher haematological responses, due to the limited body of evidence available. There is Level I evidence that, within reasonable limits of body weight, fixed doses of erythropoietic proteins can be used to treat patients with chemotherapy-induced anaemia. This analysis confirms that there are no baseline predictive factors of response to erythropoietic proteins that can be routinely used in clinical practice if functional iron deficiency or vitamin deficiency is ruled out; a low serum erythropoietin (EPO) level (only in haematological malignancies) appears to be the only predictive factor to be verified in Level I studies. Further studies are needed to investigate the value of hepcidin, c-reactive protein, and other measures as predictive factors. In these updated guidelines, we explored a new question of whether oral or intravenous iron supplementation increases the response rate to erythropoietic proteins. We found no evidence of increased response with the addition of oral iron supplementation, but there is Level II evidence of improved response to erythropoietic proteins with the addition of intravenous iron. However, the doses and schedules for intravenous iron supplementation are not yet well defined, and further studies in this area are warranted. The two major goals of erythropoietic protein therapy are prevention or elimination of transfusions and improvement of QoL. The total body of evidence shows that red blood cell (RBC) transfusion requirements are reduced following treatment with erythropoietic proteins. This analysis also confirms that QoL is significantly improved in patients with chemotherapy-induced anaemia and in those with anaemia of chronic disease following erythropoietic protein therapy, with more robust evidence now available that QoL was improved in studies investigating early intervention in cases of chemotherapy- or radiotherapy-induced anaemia. There is only indirect evidence that patients with chemotherapy-induced anaemia or anaemia of chronic disease initially classified as non-responders to standard doses proceed to respond to treatment following a dose increase. None of the studies addressed the question in a prospective, randomised fashion, and so the Taskforce does not recommend dose escalation as a general approach in all patients who are not responding. There is still insufficient data to determine the effect on survival following treatment with erythropoietic proteins in conjunction with chemotherapy or radiotherapy. Our analysis of survival endpoints in studies involving patients receiving radio(chemo)therapy found that most studies were inconclusive, with no clear link between the use of erythropoietic proteins and survival. Likewise, we found no clear link between erythropoietic therapy and other endpoints such as local tumour control, time to progression, and progression-free survival. There is no evidence that pure red cell aplasia occurs in cancer patients following treatment with erythropoietic proteins, and the fear of this condition developing should not lead to erythropoietic proteins being withheld in patients with cancer. There is Level I evidence that the risk of thromboembolic events and hypertension are slightly elevated in patients with chemotherapy-induced anaemia receiving erythropoietic proteins. Additional trials are warranted, especially to define the optimal doses and schedules of intravenous iron supplementation during erythropoietic therapy. While our review did not address cost benefit evaluations in detail, the consensus is that studies taking into account all real determinants of cost and benefit need to be performed prospectively.


Correction of the nonlinear dose response improves the viability of adenoviral vectors for gene therapy of Fabry disease.
Systemic administration of recombinant adenoviral vectors for gene therapy of chronic diseases such as Fabry disease can be limited by dose-dependent toxicity. Because administration of a high dose of Ad2/CMVHI-alpha gal encoding human alpha-galactosidase A results in expression of supraphysiological levels of the enzyme, we sought to determine whether lower doses would suffice to correct the enzyme deficiency and lysosomal storage abnormality observed in Fabry mice. Reducing the dose of Ad2/CMVHI-alpha gal by 10-fold (from 10(11) to 10(10) particles/mouse) resulted in a greater than 200-fold loss in transgene expression. In Fabry mice, the reduced expression of alpha-galactosidase A, using the lower dose of Ad2/CMVHI-alpha gal, was associated with less than optimal clearance of the accumulated glycosphingolipid (GL-3) from the affected lysosomes. It was determined that this lack of linearity in dose response was not due to an inability to deliver the recombinant viral vectors to the liver but rather to sequestration, at least in part, of the viral vectors by the Kupffer cells. This lack of correlation between dose and expression levels could be obviated by supplementing the low dose of Ad2/CMVHI-alpha gal with an unrelated adenoviral vector or by depleting the Kupffer cells before administration of Ad2/CMVHI-alpha gal. Prior removal of the Kupffer cells, using clodronate liposomes, facilitated the use of a 100-fold lower dose of Ad2/CMVHI-alpha gal (10(9) particles/mouse) to effect the nearly complete clearance of GL-3 from the affected organs of Fabry mice. These results suggest that practical strategies that minimize the interaction between the recombinant adenoviral vectors and the reticuloendothelial system (RES) may improve the therapeutic window of this vector system. In this regard, we showed that pretreatment of mice with gamma globulins also resulted in significantly enhanced adenovirus-mediated transduction and expression of alpha-galactosidase A in the liver.


Vitamin D and muscle function.
The aim of this review is to summarize current knowledge on the relation between vitamin D and muscle function. Molecular mechanisms of vitamin D action on muscle tissue have been known for many years and include genomic and non-genomic effects. Genomic effects are initiated by binding of 1,25-dihydroxyvitamin D3 (1,25(OH)2D) to its nuclear receptor, which results in changes in gene transcription of messenger RNA and subsequent protein synthesis. Non-genomic effects of vitamin D are rapid and mediated through a membrane-bound vitamin D receptor (VDR). Genetic variations in the VDR and the importance of VDR polymorphisms in the development of osteoporosis are still a matter of controversy and debate. Most recently, VDR polymorphisms have been described to affect muscle function. The skin has an enormous capacity for vitamin D production and supplies the body with 80-100% of its requirements of vitamin D. Age, latitude, time of day, season of the year and pigmentation can dramatically affect the production of vitamin D in the skin. Hypovitaminosis D is a common feature in elderly people living in northern latitudes and skin coverage has been established as an important factor leading to vitamin D deficiency. A serum 25-hydroxyvitamin D level below 50 nmol/l has been associated with increased body sway and a level below 30 nmol/l with decreased muscle strength. Changes in gait, difficulties in rising from a chair, inability to ascend stairs and diffuse muscle pain are the main clinical symptoms in osteomalacic myopathy. Calcium and vitamin D supplements together might improve neuromuscular function in elderly persons who are deficient in calcium and vitamin D. Thus 800 IU of cholecalciferol in combination with mg of elemental calcium reduces hip fractures and other non-vertebral fractures and should generally be recommended in individuals who are deficient in calcium and vitamin D. Given the strong interdependency of vitamin D deficiency, low serum calcium and high levels of parathyroid hormone, however, it is difficult to identify exact mechanisms of action.


Low-dose aripiprazole for refractory burning mouth syndrome.
We report a case of refractory burning mouth syndrome (BMS) ameliorated with low dose of aripiprazole. The patient was a 66-year-old female who had suffered from chronic burning pain in her tongue for 13 months. No abnormality associated with the burning sensation was detected in the laboratory tests and the oral findings. Considering the clinical feature and the history together, we diagnosed the burning sensation as BMS. The BMS pain was decreased by aripiprazole (powder) 1.0 mg/d, though no other antidepressants had satisfying pain relief. It could be supposed that the efficacy of aripiprazole is caused by dopamine stabilization in this case, and BMS might have a subtype that is reactive to aripiprazole. Further studies are needed to confirm the efficacy of aripiprazole for BMS.


Progression of spinal deformity in wheelchair-dependent patients with Duchenne muscular dystrophy who are not treated with steroids: coronal plane (scoliosis) and sagittal plane (kyphosis, lordosis) deformity.
We determined the frequency, rate and extent of development of scoliosis (coronal plane deformity) in wheelchair-dependent patients with Duchenne muscular dystrophy (DMD) who were not receiving steroid treatment. We also assessed kyphosis and lordosis (sagittal plane deformity). The extent of scoliosis was assessed on sitting anteroposterior (AP) spinal radiographs in 88 consecutive non-ambulatory patients with DMD. Radiographs were studied from the time the patients became wheelchair-dependent until the time of spinal fusion, or the latest assessment if surgery was not undertaken. Progression was estimated using a longitudinal mixed-model regression analysis to handle repeated measurements. Scoliosis ≥ 10° occurred in 85 of 88 patients (97%), ≥ 20° in 78 of 88 (89%) and ≥ 30° in 66 of 88 patients (75%). The fitted longitudinal model revealed that time in a wheelchair was a highly significant predictor of the magnitude of the curve, independent of the age of the patient (p < 0.001). Scoliosis developed in virtually all DMD patients not receiving steroids once they became wheelchair-dependent, and the degree of deformity deteriorated over time. In general, scoliosis increased at a constant rate, beginning at the time of wheelchair-dependency (p < 0.001). In some there was no scoliosis for as long as three years after dependency, but scoliosis then developed and increased at a constant rate. Some patients showed a rapid increase in the rate of progression of the curve after a few years - the clinical phenomenon of a rapidly collapsing curve over a few months. A sagittal plane kyphotic deformity was seen in 37 of 60 patients (62%) with appropriate radiographs, with 23 (38%) showing lumbar lordosis (16 (27%) abnormal and seven (11%) normal). This study provides a baseline to assess the effects of steroids and other forms of treatment on the natural history of scoliosis in patients with DMD, and an approach to assessing spinal deformity in the coronal and sagittal planes in wheelchair-dependent patients with other neuromuscular disorders.


In vivo effects of camostat mesilate on plasma kallikrein, plasma kininase II and renal kallikrein of man.
N,N-Dimethylcarbamoylmethyl-4-(4-guanidino-benzoyloxy)phenylacetat e methanesulfonate (camostat mesilate) is reported to be an effective inhibitor of plasma kallikrein. It was shown in vitro to inhibit not only plasma kallikrein, but also renal kallikrein and plasma kininase II. These inhibitory activities, however, were very weak. The inhibition of plasma kallikrein in human plasma was limited in time, since a rapid reactivation of plasma kallikrein was noticed when samples were incubated at room temperature. In order to establish whether camostat mesilate was able to inhibit plasma kallikrein, kininase II and renal kallikrein also in vivo the inhibitory activity of camostat mesilate on these enzymes was studied in 5 healthy volunteers. After an oral intake of a single dose of 600 mg of camostat mesilate, plasma kallikrein was inhibited significantly, while kininase II in plasma was unaffected. Renal kallikrein activity determined by urinary excretion of active kallikrein remained unchanged after camostat mesilate intake. Thus, the results demonstrate that camostat mesilate in vivo inhibits only plasma kallikrein and has no effect on the activity of kininase II or renal kallikrein.


[Examination of helminth contaminated wastewaters used for agricultural purposes in Afyonkarahisar].
This research was performed in order to examine helminth contaminated wastewaters used in agriculture in Afyonkarahisar. Ten rural areas were selected that used water for agricultural purposes from a creek contaminated with human and animal wastes. Between September 2004 and August 2005, these areas were visited once every two weeks, and a total of 240 water samples were collected. Out of untreated water samples, 32 samples (26.70%) had helminth eggs, whereas there were neither helminths nor developmental stages of helminths (p < 0.001) in treated water samples. Cestode eggs were seen in 10.00% of untreated water samples and nematode eggs were seen in 16.70%. Out of the helminth eggs, 13 (40.60%) were hookworm eggs; 6 (18.75%), Taenia spp.; 5 (5.60%), Ascaris lumbricoides; 3 (9.40%), Hymenolepis diminuta; 3 (9.40%), H. nana; and 2 (6.25%), Toxocara spp. In conclusion; the creek was found to be contaminated by helminths originating from human and animal wastes in Afyonkarahisar. This type of water presents a risk for human and animal health and contaminated waters, therefore, should be avoided in irrigation of vegetables and fruit consumed raw.


The Five "I" Framework of crash investigation: Linking investigation practices to safety reform.
Crash investigation is vital to the advancement of road safety, and it is a foundation for determining effective countermeasures and interventions for true road safety reform. However, the way in which road crash investigations are conducted substantially influences the quality of understanding and the effectiveness of responses. In crash investigations, we traditionally focus first on the question: "What caused this crash?" when it would be more efficient to ask immediately: "What could have prevented this crash?" or better yet, "What are all the ways this crash could have been prevented?" In this paper, we first explore a few common road crash investigation approaches where prosecution, retribution, or compensation are primary. We then examine investigation approaches where prevention is primary, especially investigations aimed at determining every point on the timeline preceding the crash where an intervention would have prevented the outcome, as illustrated by the Swiss Cheese Model. We draw from examples from the aviation industry, the occupational health and safety field, the U.S. National Transportation Safety Board (NTSB), and others, to identify strengths and weaknesses. We bring together good practices from several investigative approaches through the lens of diverse experiences in transportation safety and root-cause analysis to present a practical and proactive framework for road crash investigation. The Five "I" Framework provides guiding characteristics for prevention-focused safety investigations for road crashes: Immediate, In-Depth, Impartial, Independent, and Injury Prevention. The Five "I" Framework is a practical guide for investigations to move beyond crash causation to crash prevention, aligning with the Safe Systems Approach, Vision Zero, and the public health perspective. Rather than focusing primarily on on any single factor such as vehicle defect or driver error, it leads investigations to an array of countermeasures that involve collective action and systems change, and thus, to more effective road safety reform. Nevertheless, as a practical framework, it is the start (not the end) of discussions on how we can continue to move towards more multidisciplinary, collaborative, innovative, and ethical prevention efforts.