An integrative method to normalize RNA-Seq data.
Transcriptome sequencing is a powerful tool for measuring gene expression, but as well as some other technologies, various artifacts and biases affect the quantification. In order to correct some of them, several normalization approaches have emerged, differing both in the statistical strategy employed and in the type of corrected biases. However, there is no clear standard normalization method. We present a novel methodology to normalize RNA-Seq data, taking into account transcript size, GC content, and sequencing depth, which are the major quantification-related biases. In this study, we found that transcripts shorter than 600 bp have an underestimated expression level, while longer transcripts are even more overestimated that they are long. Second, it was well known that the higher the GC content (>50%), the more the transcripts are underestimated. Third, we demonstrated that the sequencing depth impacts the size bias and proposed a correction allowing the comparison of expression levels among many samples. The efficiency of our approach was then tested by comparing the correlation between normalized RNA-Seq data and qRT-PCR expression measurements. All the steps are automated in a program written in Perl and available on request. The methodology presented in this article identifies and corrects different biases that influence RNA-Seq quantification, and provides more accurate estimations of gene expression levels. This method can be applied to compare expression quantifications from many samples, but preferentially from the same tissue. In order to compare samples from different tissue, a calibration using several reference genes will be required.


Managing dental caries with atraumatic restorative treatment in children: successful experience in three Latin American countries.
To compare survival rates and cost differentials between the atraumatic restorative treatment (ART) technique and amalgams by type of dental personnel in Ecuador, Panama, and Uruguay. Children 7 to 9 years of age in rural and urban schools with at least one lesion with initial cavitated enamel caries or a dentinal lesion on a first permanent molar were selected and randomized into the ART (intervention) or amalgam (control) group. Restoration failure was evaluated at 12 and 24 months. Cooperation and pain experienced during the procedures were measured. Cumulative and incident failure of restorations at 12 and 24 months was calculated for dentists who placed ART or amalgam restorations and auxiliaries who placed ART restorations at 12 months only. The total sample comprised 1 629 children. Study groups were similar by country, gender, and geographic location. Cumulative failure rate at 12 months varied by group: dentists' amalgam, 0.9% to 5.7%; dentists' ART, 2.0% to 10.5%; and auxiliaries' ART, 5.7% to 15.8%. At 24 months, higher cumulative failures were observed for the dentists' amalgam group compared with the dentists' ART group in Ecuador and Panama but not in Uruguay. Amalgam was least likely to have the best level of cooperation and an auxiliary using ART was associated with the least pain. The cost of using the ART approach for dental caries treatment, including retreatment, was roughly half that of using amalgam without retreatment. Having auxiliary personnel perform ART will lead to treatment survival that is expected to be lower than dentists using amalgam or ART. In spite of the greater risk of failure, the rate is not unacceptable and potential cost savings are substantial.


Hypothalamic over-expression of VGF in the Siberian hamster increases energy expenditure and reduces body weight gain.
VGF (non-acronymic) was first highlighted to have a role in energy homeostasis through experiments involving dietary manipulation in mice. Fasting increased VGF mRNA in the Arc and levels were subsequently reduced upon refeeding. This anabolic role for VGF was supported by observations in a VGF null (VGF-/-) mouse and in the diet-induced and gold-thioglucose obese mice. However, this anabolic role for VGF has not been supported by a number of subsequent studies investigating the physiological effects of VGF-derived peptides. Intracerebroventricular (ICV) infusion of TLQP-21 increased resting energy expenditure and rectal temperature in mice and protected against diet-induced obesity. Similarly, ICV infusion of TLQP-21 into Siberian hamsters significantly reduced body weight, but this was due to a decrease in food intake, with no effect on energy expenditure. Subsequently NERP-2 was shown to increase food intake in rats via the orexin system, suggesting opposing roles for these VGF-derived peptides. Thus to further elucidate the role of hypothalamic VGF in the regulation of energy homeostasis we utilised a recombinant adeno-associated viral vector to over-express VGF in adult male Siberian hamsters, thus avoiding any developmental effects or associated functional compensation. Initially, hypothalamic over-expression of VGF in adult Siberian hamsters produced no effect on metabolic parameters, but by 12 weeks post-infusion hamsters had increased oxygen consumption and a tendency to increased carbon dioxide production; this attenuated body weight gain, reduced interscapular white adipose tissue and resulted in a compensatory increase in food intake. These observed changes in energy expenditure and food intake were associated with an increase in the hypothalamic contents of the VGF-derived peptides AQEE, TLQP and NERP-2. The complex phenotype of the VGF-/- mice is a likely consequence of global ablation of the gene and its derived peptides during development, as well as in the adult.


DNA-functionalized gold nanoparticles in macromolecularly crowded polymer solutions.
DNA-functionalized gold nanoparticles (AuNPs) are one of the most commonly used reagents in nanobiotechnology. They are important not only for practical applications in analytical chemistry and drug delivery, but also for fundamental understanding of nanoscience. For biological samples such as blood serum or for intracellular applications, the effects of crowded cellular proteins and nucleic acids need to be considered. The thermodynamic effect of crowding is to induce nanoparticle aggregation. But before such aggregation can take place, there might also be a depletion repulsive barrier. Polyethylene glycol (PEG) is one of the most frequently used polymers to mimic the crowded cellular environment. We show herein that while DNA-functionalized AuNPs are very stable in buffer (e.g., no PEG) and citrate-capped AuNPs are very stable in PEG, DNA-functionalized AuNPs are unstable in PEG and are easily aggregated. Although such aggregation in PEG is mediated by DNA, no sharp melting transition typical for DNA-linked AuNPs is observed. We attribute this broad melting to depletion force instead of DNA base pairing. The effects of PEG molecular weight, concentration and temperature have been studied in detail and we also find an interesting PEG phase separation and AuNP partition into the water-rich phase at high temperature.


Mucin histochemistry of mucous carcinomas of breast and colon and non-neoplastic breast epithelium.
Although epithelial mucin has been demonstrated in ordinary mammary duct carcinomas, it is formed profusely in the uncommon mucous carcinomas. A histochemical study of mucous carcinomas of the breast and colon showed that both types produce similar mucins, viz, acid and neutral, although the mammary carcinomas produce more neutral mucin and small amounts of sulphated mucins are produced by colonic tumours. Similar studies on sections of normal breast and cystic mammary dysplasia have shown that intracellular neutral mucin was present in the ducts and lobules, and that intracellular acid mucin was present within cells of the terminal ducts and lobules. It was demonstrated enzymatically that the acid mucins produced by the normal and neoplastic tissues is sialic acid. Hence it can be shown that the mucin produced by mucous carcinomas is of epithelial rather than connective tissue type. It is contended that mucous carcinoma of breast arises by metaplasia of duct cells that have already undergone malignant change and not from the lobules of the breast.


Shared effects of genetic and intrauterine and perinatal environment on the development of metabolic syndrome.
Genetic and environmental factors, including the in utero environment, contribute to Metabolic Syndrome. Exposure to high fat diet exposure in utero and lactation increases incidence of Metabolic Syndrome in offspring. Using GLUT4 heterozygous (G4+/-) mice, genetically predisposed to Type 2 Diabetes Mellitus, and wild-type littermates we demonstrate genotype specific differences to high fat in utero and lactation. High fat in utero and lactation increased adiposity and impaired insulin and glucose tolerance in both genotypes. High fat wild type offspring had increased serum glucose and PAI-1 levels and decreased adiponectin at 6 wks of age compared to control wild type. High fat G4+/- offspring had increased systolic blood pressure at 13 wks of age compared to all other groups. Potential fetal origins of adult Metabolic Syndrome were investigated. Regardless of genotype, high fat in utero decreased fetal weight and crown rump length at embryonic day 18.5 compared to control. Hepatic expression of genes involved in glycolysis, gluconeogenesis, oxidative stress and inflammation were increased with high fat in utero. Fetal serum glucose levels were decreased in high fat G4+/- compared to high fat wild type fetuses. High fat G4+/-, but not high fat wild type fetuses, had increased levels of serum cytokines (IFN-γ, MCP-1, RANTES and M-CSF) compared to control. This data demonstrates that high fat during pregnancy and lactation increases Metabolic Syndrome male offspring and that heterozygous deletion of GLUT4 augments susceptibility to increased systolic blood pressure. Fetal adaptations to high fat in utero that may predispose to Metabolic Syndrome in adulthood include changes in fetal hepatic gene expression and alterations in circulating cytokines. These results suggest that the interaction between in utero-perinatal environment and genotype plays a critical role in the developmental origin of health and disease.


Tick-susceptible Bos taurus cattle display an increased cellular response at the site of larval Rhipicephalus (Boophilus) microplus attachment, compared with tick-resistant Bos indicus cattle.
Cattle demonstrate divergent and heritable phenotypes of resistance and susceptibility to infestation with the cattle tick Rhipicephalus (Boophilus) microplus. Bos indicus cattle are generally more resistant to tick infestation than Bos taurus breeds although large variations in resistance can occur within subspecies and within breed. Increased tick resistance has been previously associated with an intense hypersensitivity response in B. taurus breeds; however, the mechanism by which highly resistant B. indicus cattle acquire and sustain high levels of tick resistance remains to be elucidated. Using the commercially available Affymetrix microarray gene expression platform, together with histological examination of the larval attachment site, this study aimed to describe those processes responsible for high levels of tick resistance in Brahman (B. indicus) cattle that differ from those in low-resistance Holstein-Friesian (B. taurus) cattle. We found that genes involved in inflammatory processes and immune responsiveness to infestation by ticks, although up-regulated in tick-infested Holstein-Friesian cattle, were not up-regulated in Brahman cattle. In contrast, genes encoding constituents of the extracellular matrix were up-regulated in Brahmans. Furthermore, the susceptible Holstein-Friesian animals displayed a much greater cellular inflammatory response at the site of larval R. microplus attachment compared with the tick-resistant Brahman cattle.


Repeatability Assessment of Intravascular Polarimetry in Patients.
Intravascular polarimetry with polarization sensitive optical frequency domain imaging (PS-OFDI) measures polarization properties of the vessel wall and offers characterization of coronary atherosclerotic lesions beyond the cross-sectional image of arterial microstructure available to conventional OFDI. A previous study of intravascular polarimetry in cadaveric human coronary arteries found that tissue birefringence and depolarization provide valuable insight into key features of atherosclerotic plaques. In addition to various tissue components, catheter and sample motion can also influence the polarization of near infrared light as used by PS-OFDI. This paper aimed to evaluate the robustness and repeatability of imaging tissue birefringence and depolarization in a clinical setting. 30 patients scheduled for percutaneous coronary intervention at the Erasmus Medical Center underwent repeated PS-OFDI pullback imaging, using commercial imaging catheters in combination with a custom-built PS-OFDI console. We identified 274 matching cross sections among the repeat pullbacks to evaluate the reproducibility of the conventional backscatter intensity, the birefringence, and the depolarization signals at each spatial location across the vessel wall. Bland-Altman analysis revealed best agreement for the birefringence measurements, followed by backscatter intensity, and depolarization, when limiting the analysis to areas of meaningful birefringence. Pearson correlation analysis confirmed highest correlation for birefringence (0.86), preceding backscatter intensity (0.83), and depolarization (0.78). Our results demonstrate that intravascular polarimetry generates robust maps of tissue birefringence and depolarization in a clinical setting. This outcome motivates the use of intravascular polarimetry for future clinical studies that investigate polarization properties of arterial atherosclerosis.


Treatment and compliance with virtual reality and anaglyph-based training programs for convergence insufficiency.
Convergence insufficiency may be treated by visual exercises designed to increase convergence while maintaining single, clear, binocular vision. However, compliance with treatment is problematic, as patients often cease treatment when symptoms start to improve and before treatment is complete. The purpose of the present study was to assess the feasibility of using gamification of vision training to: (a) treat convergence insufficiency; and (b) improve compliance to treatment in comparison to a conventional treatment over a six-week treatment period. Two interventions, anaglyphs and a virtual reality game of Snakes, were evaluated for their effectiveness in treating adults with convergence insufficiency. The prescribed training regimen was 20 minutes, three times per week for six weeks. Vision was assessed before and after the treatment period. Participants also filled in the Core Elements of the Gaming Experience Questionnaire to gauge impact of game design on compliance. Eighteen participants (mean age 20.8 ± 1.8 years) met the inclusion criteria for convergence insufficiency and nine participants were randomly assigned to each intervention. Repeated measures analysis of variance showed a significant effect of visit for near point of convergence (F1,16 = 38.32, p < 0.0001), near positive fusional reserves break (F1,16 = 21.94, p < 0.0001) and recovery (F1,16 = 26.87, p < 0.0001), but not of intervention type. Total time played was significantly longer for the virtual reality Snake Game than the anaglyph intervention (p < 0.0001), which translated to mean compliance of 82 per cent and 51 per cent respectively. Gamification of vision training in a virtual reality environment is feasible and associated with increased compliance, hence may be a useful strategy to treat convergence insufficiency.


In vitro modelling of alveolar repair at the air-liquid interface using alveolar epithelial cells derived from human induced pluripotent stem cells.
Research on acute and chronic lung diseases would greatly benefit from reproducible availability of alveolar epithelial cells (AEC). Primary alveolar epithelial cells can be derived from human lung tissue but the quality of these cells is highly donor dependent. Here, we demonstrated that culture of EpCAM+ cells derived from human induced pluripotent stem cells (hiPSC) at the physiological air-liquid interface (ALI) resulted in type 2 AEC-like cells (iAEC2) with alveolar characteristics. iAEC2 cells expressed native AEC2 markers (surfactant proteins and LPCAT-1) and contained lamellar bodies. ALI-iAEC2 were used to study alveolar repair over a period of 2 weeks following mechanical wounding of the cultures and the responses were compared with those obtained using primary AEC2 (pAEC2) isolated from resected lung tissue. Addition of the Wnt/β-catenin activator CHIR99021 reduced wound closure in the iAEC2 cultures but not pAEC2 cultures. This was accompanied by decreased surfactant protein expression and accumulation of podoplanin-positive cells at the wound edge. These results demonstrated the feasibility of studying alveolar repair using hiPSC-AEC2 cultured at the ALI and indicated that this model can be used in the future to study modulation of alveolar repair by (pharmaceutical) compounds.


Conformational optimization with natural degrees of freedom: a novel stochastic chain closure algorithm.
The present article introduces a set of novel methods that facilitate the use of "natural moves" or arbitrary degrees of freedom that can give rise to collective rearrangements in the structure of biological macromolecules. While such "natural moves" may spoil the stereochemistry and even break the bonded chain at multiple locations, our new method restores the correct chain geometry by adjusting bond and torsion angles in an arbitrary defined molten zone. This is done by successive stages of partial closure that propagate the location of the chain break backwards along the chain. At the end of these stages, the size of the chain break is generally reduced so much that it can be repaired by adjusting the position of a single atom. Our chain closure method is efficient with a computational complexity of O(N(d)), where N(d) is the number of degrees of freedom used to repair the chain break. The new method facilitates the use of arbitrary degrees of freedom including the "natural" degrees of freedom inferred from analyzing experimental (X-ray crystallography and nuclear magnetic resonance [NMR]) structures of nucleic acids and proteins. In terms of its ability to generate large conformational moves and its effectiveness in locating low energy states, the new method is robust and computationally efficient.


The Postprandial Appearance of Features of Cardiometabolic Risk: Acute Induction and Prevention by Nutrients and Other Dietary Substances.
The purpose of this review is to provide an overview of diets, food, and food components that affect postprandial inflammation, endothelial function, and oxidative stress, which are related to cardiometabolic risk. A high-energy meal, rich in saturated fat and sugars, induces the transient appearance of a series of metabolic, signaling and physiological dysregulations or dysfunctions, including oxidative stress, low-grade inflammation, and endothelial dysfunction, which are directly related to the amplitude of postprandial plasma triglycerides and glucose. Low-grade inflammation and endothelial dysfunction are also known to cluster together with insulin resistance, a third risk factor for cardiovascular diseases (CVD) and type-II diabetes, thus making a considerable contribution to cardiometabolic risk. Because of the marked relevance of the postprandial model to nutritional pathophysiology, many studies have investigated whether adding various nutrients and other substances to such a challenge meal might mitigate the onset of these adverse effects. Some foods (e.g., nuts, berries, and citrus), nutrients (e.g., l-arginine), and other substances (various polyphenols) have been widely studied. Reports of favorable effects in the postprandial state have concerned plasma markers for systemic or vascular pro-inflammatory conditions, the activation of inflammatory pathways in plasma monocytes, vascular endothelial function (mostly assessed using physiological criteria), and postprandial oxidative stress. Although the literature is fragmented, this topic warrants further study using multiple endpoints and markers to investigate whether the interesting candidates identified might prevent or limit the postprandial appearance of critical features of cardiometabolic risk.


Pharmacologic management of osteoarthritis-related pain in older adults: a review shows that many drug therapies provide small-to-modest pain relief.
Because pain is a common and debilitating symptom of osteoarthritis in older adults, the authors reviewed data on the efficacy and safety of commonly used oral, topical, and intraarticular drug therapies in this population. A search of several databases found that most studies have focused on knee osteoarthritis and reported only short-term outcomes. Also, treatment efficacy was found to vary by drug class; the smallest effect was observed with acetaminophen and the largest with opioids and viscosupplements.Acetaminophen and topical agents had the best safety profiles, whereas oral nonsteroidal antiinflammatory drugs and opioids had the worst. Little data were available on patients ages 75 years old and older and on patients from diverse racial and ethnic groups. Most drug therapies gave mild-to-moderate pain relief; their long-term safety and efficacy and their effects in diverse populations (particularly older adults) remain undetermined.


Elicitation of structure-specific antibodies by epitope scaffolds.
Elicitation of antibodies against targets that are immunorecessive, cryptic, or transient in their native context has been a challenge for vaccine design. Here we demonstrate the elicitation of structure-specific antibodies against the HIV-1 gp41 epitope of the broadly neutralizing antibody 2F5. This conformationally flexible region of gp41 assumes mostly helical conformations but adopts a kinked, extended structure when bound by antibody 2F5. Computational techniques were employed to transplant the 2F5 epitope into select acceptor scaffolds. The resultant "2F5-epitope scaffolds" possessed nanomolar affinity for antibody 2F5 and a range of epitope flexibilities and antigenic specificities. Crystallographic characterization of the epitope scaffold with highest affinity and antigenic discrimination confirmed good to near perfect attainment of the target conformation for the gp41 molecular graft in free and 2F5-bound states, respectively. Animals immunized with 2F5-epitope scaffolds showed levels of graft-specific immune responses that correlated with graft flexibility (p < 0.04), while antibody responses against the graft-as dissected residue-by-residue with alanine substitutions-resembled more closely those of 2F5 than sera elicited with flexible or cyclized peptides, a resemblance heightened by heterologous prime-boost. Lastly, crystal structures of a gp41 peptide in complex with monoclonal antibodies elicited by the 2F5-epitope scaffolds revealed that the elicited antibodies induce gp41 to assume its 2F5-recognized shape. Epitope scaffolds thus provide a means to elicit antibodies that recognize a predetermined target shape and sequence, even if that shape is transient in nature, and a means by which to dissect factors influencing such elicitation.


Primary appendicular hemangiosarcoma and telangiectatic osteosarcoma in 70 dogs: A Veterinary Society of Surgical Oncology retrospective study.
To define and compare clinical characteristics of canine primary appendicular hemangiosarcoma (HSA) and telangiectatic osteosarcoma (tOSA), including signalment, presentation, response to treatment, and prognosis. Multi-institutional retrospective study. Seventy dogs with primary appendicular HSA or tOSA. Patient data were obtained from institutions' medical records. Immunohistochemistry was applied to archived tissues to establish tumor type. Patient characteristics, treatment responses, and outcomes were described and compared by tumor type. Forty-one HSA and 29 tOSA were identified. Dogs with HSA were more likely than dogs with tOSA to be male and have hind limb tumors; 78% of HSA occurred in hind limbs, particularly the tibia. Dogs with tOSA weighed a median of 9.9 kg (95% CI 4.6-15.3) more than dogs with HSA. Most dogs received antineoplastic treatment, predominantly amputation with or without adjuvant chemotherapy. Overall survival with local treatment and chemotherapy was 299 days (95% CI 123-750) for HSA and 213 days (95% CI 77-310) for tOSA. Younger age and more aggressive treatment were associated with longer survival in dogs with HSA but not tOSA. One-year survival rates did not differ between dogs with HSA (28%) and those with tOSA (7%). Distinct clinical features were identified between HSA and tOSA in this population. Both tumors were aggressive, with a high incidence of pulmonary metastases. However, local treatment combined with chemotherapy led to an average survival 7 months for tOSA and 10 months for HSA. HSA should be considered as a differential in dogs with aggressive lytic bone lesions, particularly medium-sized dogs with tibial lesions. HSA has a unique clinical presentation but similar therapeutic response and outcome to OSA. Amputation and chemotherapy appear to prolong survival in some dogs with HSA and tOSA.


Whole-brain adaptive 70-kVp perfusion imaging with variable and extended sampling improves quality and consistency while reducing dose.
Despite common use of CTP to assess cerebral hemodynamics in the setting of ischemia, concerns over radiation exposure remain. Our aim was to evaluate the efficacy of an adaptive 70-kVp (peak) whole-brain CTP protocol with variable sampling intervals and extended duration against an established fixed-sampling, limited-period protocol at 80 kVp. A retrospective analysis of 37 patients with stroke scanned with conventional (n = 17) and variant-protocol (n = 20) whole-brain CTP was performed. We compared radiation dose, parametric map quality, and consistency of full-contrast circulation capture between a modified 70-kVp protocol, with 20 whole-brain passes at variable sampling intervals over an extended sampling period, and a conventional 80-kVp CTP examination with 24 passes at fixed-sampling intervals and a more limited scanning window. Mann-Whitney U test analysis was used to compare both protocols. The 70-kVp CTP scan provided superior image quality at a 45% lower CT dose index volume and 13% lower dose-length product/effective dose compared with the conventional 80-kVp scan. With respect to the consistency of contrast-passage capture, 95% of the adaptive, extended protocol continued through the venous return to baseline, compared with only 47% by using the conventional limited-length protocol. Rapid sampling during the critical arterial arrival and washout period was accomplished in nearly 95% with both the variable and fixed-sampling-interval protocols. Seventy-kilovolt (peak) CTP with variable and extended sampling produces improved image quality at lower radiation doses with greater consistency of full contrast passage capture.


Aquatic Orthopaedic Injuries.
Extremity injuries sustained in aquatic environments require unique considerations compared with injuries sustained on land. Knowledge of these considerations is becoming more important as aquatic recreational activities increase in popularity. Aquatic injuries may occur through mechanical contact with a variety of different objects or surfaces, such as a recreational device or watercraft part, or may occur through contact with marine animals. Marine animal injuries can be further categorized into bites, stings, or blunt contact, as well as venomous or nonvenomous, distinctions that should be used to guide clinical management. Numerous instances of retained foreign bodies after marine animal stings exist, which can result in infection and prolonged envenomization; thus, radiographic examination should be routinely performed in aquatic sting injuries to prevent these harmful sequelae. Any aquatic injury resulting in an open wound has an increased risk for infection, and prophylactic antibiotics must be given with consideration for the unique microbiologic flora of the aquatic environment.


Hereditary nephritis in the bull terrier: evidence for inheritance by an autosomal dominant gene.
A high prevalence of renal failure has been reported in bull terriers in Australia. The pattern of inheritance was analysed in a family of 33 bull terriers in which 10 dogs had renal disease manifested by proteinuria, ultrastructural abnormalities in the glomerular basement membrane, renal failure, or 'end stage' kidneys. The presence of at least one affected parent for each affected offspring, the approximately equal male/female ratio and the apparent absence of 'generation-skipping', strongly supported an autosomal dominant mode of inheritance, assuming a fully penetrant single major gene locus. Further evidence was not compatible with either an autosomal recessive or X-linked inheritance pattern. This contrasts with the X-linked inheritance shown in Alport's-type human hereditary nephritis and hereditary glomerulopathy in the samoyeds. Hereditary nephritis in the bull terrier should be a useful model for non-Alport's-type human hereditary nephritis, which is also reported to have an autosomal dominant inheritance pattern.


Dual cationic-anionic profiling of metabolites in a single identified cell in a live Xenopus laevis embryo by microprobe CE-ESI-MS.
In situ capillary microsampling with capillary electrophoresis (CE) electrospray ionization (ESI) mass spectrometry (MS) enabled the characterization of cationic metabolites in single cells in complex tissues and organisms. For deeper coverage of the metabolome and metabolic networks, analytical approaches are needed that provide complementary detection for anionic metabolites, ideally using the same instrumentation. Described here is one such approach that enables sequential cationic and anionic (dual) analysis of metabolites in the same identified cell in a live vertebrate embryo. A calibrated volume was microaspirated from the animal-ventral cell in a live 8-cell embryo of Xenopus laevis, and cationic and anionic metabolites were one-pot microextracted from the aspirate, followed by CE-ESI-MS analysis of the same extract. A laboratory-built CE-ESI interface was reconfigured to enable dual cationic-anionic analysis with ∼5-10 nM (50-100 amol) lower limit of detection and a capability for quantification. To provide robust separation and efficient ion generation, the CE-ESI interface was enclosed in a nitrogen gas filled chamber, and the operational parameters were optimized for the cone-jet spraying regime in both the positive and negative ion mode. A total of ∼250 cationic and ∼200 anionic molecular features were detected from the cell between m/z 50-550, including 60 and 24 identified metabolites, respectively. With only 11 metabolites identified mutually, the duplexed approach yielded complementary information on metabolites produced in the cell, which in turn deepened network coverage for several metabolic pathways. With scalability to smaller cells and adaptability to other types of tissues and organisms, dual cationic-anionic detection with in situ microprobe CE-ESI-MS opens a door to better understand cell metabolism.


Efficacy, safety and tolerability of linezolid containing regimens in treating MDR-TB and XDR-TB: systematic review and meta-analysis.
Linezolid is used off-label to treat multidrug-resistant tuberculosis (MDR-TB) in absence of systematic evidence. We performed a systematic review and meta-analysis on efficacy, safety and tolerability of linezolid-containing regimes based on individual data analysis. 12 studies (11 countries from three continents) reporting complete information on safety, tolerability, efficacy of linezolid-containing regimes in treating MDR-TB cases were identified based on Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Meta-analysis was performed using the individual data of 121 patients with a definite treatment outcome (cure, completion, death or failure). Most MDR-TB cases achieved sputum smear (86 (92.5%) out of 93) and culture (100 (93.5%) out of 107) conversion after treatment with individualised regimens containing linezolid (median (inter-quartile range) times for smear and culture conversions were 43.5 (21-90) and 61 (29-119) days, respectively) and 99 (81.8%) out of 121 patients were successfully treated. No significant differences were detected in the subgroup efficacy analysis (daily linezolid dosage ≤ 600 mg versus >600 mg). Adverse events were observed in 63 (58.9%) out of 107 patients, of which 54 (68.4%) out of 79 were major adverse events that included anaemia (38.1%), peripheral neuropathy (47.1%), gastro-intestinal disorders (16.7%), optic neuritis (13.2%) and thrombocytopenia (11.8%). The proportion of adverse events was significantly higher when the linezolid daily dosage exceeded 600 mg. The study results suggest an excellent efficacy but also the necessity of caution in the prescription of linezolid.


Changes in erectile response to repeated audiovisual sexual stimulation.
To determine changes in erectile response to repeated audiovisual sexual stimulation in patients with psychogenic impotence and normal men. The same erotic video was shown to 45 men (20 patients with psychogenic impotence and 25 normal men) for 3 consecutive days. Their erectile responses during the audiovisual stimulation were monitored using RigiScan (Dacomed, Minneapolis, Minn., USA), and the maximal rigidity of erection, sustained for more than 5 min, was measured. The rigidities on the first, second and third days were comparatively analyzed. Whether previous multiple viewings of the erotic movie and previous exposure to the same or a similar movie influenced the erectile response were also evaluated. Rigidity on the third day was significantly decreased compared to that on the first day in both patients with psychogenic impotence and normal controls (p < 0.05), regardless of the frequency of previous viewings and the previous exposure to a similar movie. Rigidity on the first day was significantly lower in the group with multiple viewings than in the group with fewer viewings (p < 0.05). Real-time monitoring of penile erection during audiovisual sexual stimulation may result in false-negative responses when patients are repeatedly exposed to the stimulation.


Tocotrienols confer resistance to ischemia in hypercholesterolemic hearts: insight with genomics.
Most clinical trials with vitamin E could not lower cholesterol and thus, have been deemed unsuccessful. Recently, tocotrienols, isomers of vitamin E have been found to lower LDL levels. To explore if tocotrienols could be the drug target for vitamin E, rabbits were kept on cholesterol diet for 60 days supplemented with tocotrienol-α, tocotrienol-δ, and tocotrienol-γ for the last 30 days. The serum cholesterol levels (in mmol/l) were 24.4 (tocotrienol-α), 34.9 (tocotrienol-δ), 19.8 (tocotrienol-γ) vs. 39.7 (control). Left ventricular function including aortic flow and developed pressure exhibited significantly improved recovery with tocotrienol-γ and -α, but not with tocotrienol-δ. The myocardial infarct size showed a similar pattern: 33% (tocotrienol-α), 23% (tocotrienol-γ), and 47% (tocotrienol-δ). To examine the molecular mechanisms of cardioprotective effects, gene expression profile was determined using Atlas 1.2/1.2II followed by determination of gene profiles using PedQuest 8.3 software. Based on genomic profiles, the following cholesterol-related proteins were examined: FABP, TGF-β (cholesterol suppresses TGF-β), ET-1 (increased by hypercholesterolemia), SPOT 14 (linked with hypercholesterolemia), and matrix metalloproteinase (MMP) 2 and MMP9 (cholesterol regulates MMP2 and MMP9 expression) in the heart. Consistent with the cardioprotective effects of tocotrienol-α and -γ, these two isomers reduced ET-1, decreased MMP2 and MM9, increased TGF-β and reduced SPOT 14, while tocotrienol-δ had no effects. The results of the present study demonstrate that the two isomers of tocotrienols, α and γ, render the hypercholesterolemic hearts resistant to ischemic reperfusion injury by lowering several hypercholesterolemic proteins including MMP2, MMP9, ET-1, and SPOT 14 and upregulating TGF-β.


Long-term outcomes of septal reduction for obstructive hypertrophic cardiomyopathy.
Surgical myectomy and alcohol septal ablation (ASA) aim to decrease left ventricular outflow tract (LVOT) gradient in hypertrophic cardiomyopathy (HCM). Outcome of myectomy beyond 10 years has rarely been described. We describe 20 years of follow-up of surgical myectomy and 5 years of follow-up for ASA performed for obstructive HCM. We studied 171 patients who underwent myectomy for symptomatic LVOT obstruction between 1972 and 2006. In addition, we studied 52 patients who underwent ASA for the same indication and who declined surgery. Follow-up of New York Heart Association (NYHA) functional class, echocardiographic data, and vital status were obtained from patient records. Mortality rates were compared with expected mortality rates of age- and sex-matched populations. Surgical myectomy improved NYHA class (2.74±0.65 to 1.54±0.74, p<0.001), reduced resting gradient (67.4±43.4mmHg to 11.2±16.4mmHg, p<0.001), and inducible LVOT gradient (98.1±34.7mmHg to 33.6±34.9mmHg, p<0.001). Similarly, ASA improved functional class (2.99±0.35 to 1.5±0.74, p<0.001), resting gradient (67.1±26.9mmHg to 23.9±29.4mmHg, p<0.001) and provoked gradient (104.4±34.9mmHg to 35.5±38.6mmHg, p<0.001). Survival after myectomy at 5, 10, 15, and 20 years of follow-up was 92.9%, 81.1%, 68.9%, and 47.5%, respectively. Of note, long-term survival after myectomy was lower than for the general population [standardized mortality ratio (SMR)=1.40, p<0.005], but still compared favorably with historical data from non-operated HCM patients. Survival after ASA at 2 and 5 years was 97.8% and 94.7%, respectively. Short-term (5 year) survival after ASA (SMR=0.61, p=0.48) was comparable to that of the general population. Long-term follow-up of septal reduction strategies in obstructive HCM reveals that surgical myectomy and ASA are effective for symptom relief and LVOT gradient reduction and are associated with favorable survival. While overall prognosis for the community HCM population is similar to the general population, the need for surgical myectomy may identify a sub-group with poorer long-term prognosis. We await long-term outcomes of more extensive myectomy approaches adopted in the past 10 years at major institutions.


Diversification and extinction in the history of life.
Analysis of the fossil record of microbes, algae, fungi, protists, plants, and animals shows that the diversity of both marine and continental life increased exponentially since the end of the Precambrian. This diversification was interrupted by mass extinctions, the largest of which occurred in the Early Cambrian, Late Ordovician, Late Devonian, Late Permian, Early Triassic, Late Triassic, and end-Cretaceous. Most of these extinctions were experienced by both marine and continental organisms. As for the periodicity of mass extinctions, no support was found: Seven mass extinction peaks in the last 250 million years are spaced 20 to 60 million years apart.


Disease-related phenotypes in a Drosophila model of hereditary spastic paraplegia are ameliorated by treatment with vinblastine.
Hereditary spastic paraplegias (HSPs) are a group of neurodegenerative diseases characterized by progressive weakness and spasticity of the lower limbs. Dominant mutations in the human SPG4 gene, encoding spastin, are responsible for the most frequent form of HSP. Spastin is an ATPase that binds microtubules and localizes to the spindle pole and distal axon in mammalian cell lines. Furthermore, its Drosophila homolog, Drosophila spastin (Dspastin), has been recently shown to regulate microtubule stability and synaptic function at the Drosophila larval neuromuscular junction. Here we report the generation of a spastin-linked HSP animal model and show that in Drosophila, neural knockdown of Dspastin and, conversely, neural overexpression of Dspastin containing a conserved pathogenic mutation both recapitulate some phenotypic aspects of the human disease, including adult onset, locomotor impairment, and neurodegeneration. At the subcellular level, neuronal expression of both Dspastin RNA interference and mutant Dspastin cause an excessive stabilization of microtubules in the neuromuscular junction synapse. In addition, we provide evidence that administration of the microtubule targeting drug vinblastine significantly attenuates these phenotypes in vivo. Our findings demonstrate that loss of spastin function elicits HSP-like phenotypes in Drosophila, provide novel insights into the molecular mechanism of spastin mutations, and raise the possibility that therapy with Vinca alkaloids may be efficacious in spastin-associated HSP and other disorders related to microtubule dysfunction.


Horopters--definition and construction.
The feature of Horopter was studied allready since the arabic and persian school, where Aguilonius defined it in 1613 for the first time. From those times til now, horopter was investigated as a geometrical feature, but also as a physiological feature of single vision. In general, there is the geometrical or theoretical horopter (Vieth, G. 1818, Muller, J. 1823) and the empirical horopter (Wheatstone, C. 1838, Panum, P L. 1858). Helmholtz includes cyclo-rotation of the eye and though geometrically defines the horopter as a "twisted cubic" fenomena, which accept also Schreiber, K.M. (2006). Our approach is geometrically and includes trigonometrical analysis of the visual lines and fixation points in space, but including the eye accommodation because the horopter plane in space is determined with the convergence angle of the bulbus and the accommodation sharpness of the eye near the fixation point and the whole presenting retina in the horopter space. We get the horopter with the presentation of both retinas in space, shaped as two spherical planes (calots), two semi-spheres with a common center of fixation. The width of their spacing which is the Panum's fusional area known as confusion of accommodation corresponds to the convergence angle of both bulbuae. If the fixation point is nearer, the Panum's fusional area is wider and hence the larger the disparation of imagies on the retina. The authors have mathematically estimated the radius of the horopter planes as: R = PD/2cosa.


Effect of initial placement level and wall thickness on maintenance of the marginal bone level in implants with a conical implant-abutment interface: a 5-year retrospective study.
Implant wall thickness and the height of the implant-abutment interface are known as factors that affect the distribution of stress on the marginal bone around the implant. The goal of this study was to evaluate the long-term effects of supracrestal implant placement and implant wall thickness on maintenance of the marginal bone level. In this retrospective study, 101 patients with a single implant were divided into the following 4 groups according to the thickness of the implant wall and the initial implant placement level immediately after surgery: 0.75 mm wall thickness, epicrestal position; 0.95 mm wall thickness, epicrestal position; 0.75 mm wall thickness, supracrestal position; 0.95 mm wall thickness, supracrestal position. The marginal bone level change was assessed 1 day after implant placement, immediately after functional loading, and 1 to 5 years after prosthesis delivery. To compare the marginal bone level change, repeated-measures analysis of variance was used to evaluate the statistical significance of differences within groups and between groups over time. Pearson correlation coefficients were also calculated to analyze the correlation between implant placement level and bone loss. Statistically significant differences in bone loss among the 4 groups (P<0.01) and within each group over time (P<0.01) were observed. There was no significant difference between the groups with a wall thickness of 0.75 mm and 0.95 mm. In a multiple comparison, the groups with a supracrestal placement level showed greater bone loss than the epicrestal placement groups. In addition, a significant correlation between implant placement level and marginal bone loss was observed. The degree of bone resorption was significantly higher for implants with a supracrestal placement compared to those with an epicrestal placement.


Critical Hypophosphatemia in a Special Operations Combat Dive Candidate: A Case Report.
In contrast to shallow water (hypoxic) blackout and swimming-induced pulmonary edema (SIPE), acute electrolyte disturbance secondary to acute respiratory alkalosis is not considered a common Combat Swimmer injury but has the potential to be life-threatening. We present the case of a 28-year-old Special Operations Dive Candidate who presented to the Emergency Department after a near-drowning incident with altered mental status, generalized weakness, respiratory distress, and tetany. He was found to have severe symptomatic hypophosphatemia (1.00mg/dL) and mild hypocalcemia secondary to intentional hyperventilation between subsurface "cross-overs," causing subsequent acute respiratory alkalosis. This is a unique presentation of a common electrolyte abnormality in a highly specialized population that is self-limiting when caused by acute respiratory alkalosis but poses a significant danger to Combat Swimmers if rescue personnel are not able to respond quickly.


Transcriptomic Analysis of Gill and Kidney from Asian Seabass (Lates calcarifer) Acclimated to Different Salinities Reveals Pathways Involved with Euryhalinity.
Asian seabass (or commonly known as barramundi), Lates calcarifer, is a bony euryhaline teleost from the Family Latidae, inhabiting nearshore, estuarine, and marine connected freshwaters throughout the tropical Indo-West Pacific region. The species is catadromous, whereby adults spawn in salinities between 28 and 34 ppt at the mouth of estuaries, with resultant juveniles usually moving into brackish and freshwater systems to mature, before returning to the sea to spawn again as adults. The species lives in both marine and freshwater habitats and can move quickly between the two; thus, the species' ability to tolerate changes in salinity makes it a good candidate for studying the salinity acclimation response in teleosts. In this study, the transcriptome of two major osmoregulatory organs (gills and kidneys) of young juvenile Asian seabass reared in freshwater and seawater were compared. The euryhaline nature of Asian seabass was found to be highly pliable and the moldability of the trait was further confirmed by histological analyses of gills and kidneys. Differences in major expression pathways were observed, with differentially expressed genes including those related to osmoregulation, tissue/organ morphogenesis, and cell volume regulation as central to the osmo-adaptive response. Additionally, genes coding for mucins were upregulated specifically under saline conditions, whereas several genes important for growth and development, as well as circadian entrainment were specifically enriched in fish reared in freshwater. Routing of the circadian rhythm mediated by salinity changes could be the initial step in salinity acclimation and possibly migration in euryhaline fish species such as the Asian seabass.


Phase II study of a biweekly regimen of vinorelbine and cisplatin in advanced non-small cell lung cancer.
Many novel agents, including vinorelbine, gemcitabine, paclitaxel and docetaxel, have been administrated in combination with cisplatin to patients with advanced non-small cell lung cancer (NSCLC). Of these drugs, vinorelbine is reported to have a high response rate and acceptable toxicity level. In an attempt to increase treatment activity, a biweekly regimen using vinorelbine and cisplatin was designed. From March 2001 to July 2003, 43 patients with NSCLC, who met the selection criteria, were enrolled. Of the 43 patients, 28 were male and 15 were female. All of them had their diagnosis confirmed histologically and were in an advanced stage, i.e., stage IIIB with pleural effusion or stage IV. Vinorelbine 30 or 35 mg/m2 and cisplatin 50 mg/m2 were given intravenously every 2 weeks. Of the 43 assessable patients, 11 achieved a Partial Response (PR) and 13 had a Stable Disease (SD). Overall response was 25.6% (95% CI 12.0%-39.2%). Median survival was 9.0 months (95% CI: 6.2-11.8) and the 1-year survival rate was 32.6%. Median time to disease progression was 3.9 months (95% CI 2.4-5.4 months). The major hematological toxicity was neutropenia. Seven patients (16.3%) developed grade 3 neutropenia and 17 patients (39.5%) developed grade 4 neutropenia. Eight patients developed febrile neutropenia, 4 patients had confirmed sepsis, 2 of which died due to sepsis. One patient had grade 3 thrombocytopenia. Six patients (7%) developed severe anemia. Ten patients (23.3%) had grade 3/4 nausea and vomiting. Only 2 patients developed grade 3 neuropathy. This biweekly regimen of vinorelbine and cisplatin is effective against advanced NSCLC. Due to the high incidence of neutropenia, this regimen did not improve therapeutic efficacy and its dose intensity is less than that of a conventional schedule.


[Current trends in using PET radiopharmaceuticals for diagnostics in oncology].
Nuclear medicine is an important field of modern medicine, particularly thanks to its role in in vivo imaging of important processes in human organism. This is possible thanks to the use of radiopharmaceuticals, specific substances labeled by radioactive nuclide, its distribution in the body can be visualized by specialized scanners and, based on the knowledge of physiological patterns, dia-gnosis can be determined. Positron emission tomography (PET) is a modern and in many ways indispensable method of nuclear medicine. The spectrum of radiopharmaceuticals available in recent years is broadening thanks to a coordinated effort of manufacturers of synthesis equipment, chemists and potential users - physicians. This review focuses on the development in the PET radiopharmaceutical field in the last five years, with an emphasis on oncological applications of PET.


An evaluation of the effectiveness of recruitment methods: the staying well after depression randomized controlled trial.
Randomized controlled trials (RCTs) are widely accepted as being the most efficient way of investigating the efficacy of psychological therapies. However, researchers conducting RCTs commonly report difficulties in recruiting an adequate sample within planned timescales. In an effort to overcome recruitment difficulties, researchers often are forced to expand their recruitment criteria or extend the recruitment phase, thus increasing costs and delaying publication of results. Research investigating the effectiveness of recruitment strategies is limited, and trials often fail to report sufficient details about the recruitment sources and resources utilized. We examined the efficacy of strategies implemented during the Staying Well after Depression RCT in Oxford to recruit participants with a history of recurrent depression. We describe eight recruitment methods utilized and two further sources not initiated by the research team and examine their efficacy in terms of (1) the return, including the number of potential participants who contacted the trial and the number who were randomized into the trial; (2) cost-effectiveness, comprising direct financial cost and manpower for initial contacts and randomized participants; and (3) comparison of sociodemographic characteristics of individuals recruited from different sources. Poster advertising, web-based advertising, and mental health worker referrals were the cheapest methods per randomized participant; however, the ratio of randomized participants to initial contacts differed markedly per source. Advertising online, via posters, and on a local radio station were the most cost-effective recruitment methods for soliciting participants who subsequently were randomized into the trial. Advertising across many sources (saturation) was found to be important. It may not be feasible to employ all the recruitment methods used in this trial to obtain participation from other populations, such as those currently unwell, or in other geographical locations. Recruitment source was unavailable for participants who could not be reached after the initial contact. Thus, it is possible that the efficiency of certain methods of recruitment was poorer than estimated. Efficacy and costs of other recruitment initiatives, such as providing travel expenses to the in-person eligibility assessment and making follow-up telephone calls to candidates who contacted the recruitment team but could not be screened promptly, were not analysed. Website advertising resulted in the highest number of randomized participants and was the second cheapest method of recruiting. Future research should evaluate the effectiveness of recruitment strategies for other samples to contribute to a comprehensive base of knowledge for future RCTs.


Breast cancer cells stimulate neutrophils to produce oncostatin M: potential implications for tumor progression.
Tumor-associated and tumor-infiltrating neutrophils (TAN) and macrophages (TAM) can account for as much as 50% of the total tumor mass in invasive breast carcinomas. It is thought that tumors secrete factors that elicit a wound-repair response from TAMs and TANs and that this response inadvertently stimulates tumor progression. Oncostatin M is a pleiotropic cytokine belonging to the interleukin-6 family that is expressed by several cell types including activated human T lymphocytes, macrophages, and neutrophils. Whereas oncostatin M can inhibit the proliferation of breast cancer cells in vitro, recent studies suggest that oncostatin M may promote tumor progression by enhancing angiogenesis and metastasis. In addition, neutrophils can be stimulated to synthesize and rapidly release large quantities of oncostatin M. In this article, we show that human neutrophils secrete oncostatin M when cocultured with MDA-MB-231 and T47D human breast cancer cells. Neutrophils isolated from whole blood or breast cancer cells alone express little oncostatin M by immunocytochemistry and ELISA, but neutrophils express and release high levels of oncostatin M when they are cocultured with breast cancer cells. In addition, we show that granulocyte-macrophage colony-stimulating factor produced by breast cancer cells and cell-cell contact are both necessary for the release of oncostatin M from neutrophils. Importantly, neutrophil-derived oncostatin M induces vascular endothelial growth factor from breast cancer cells in coculture and increases breast cancer cell detachment and invasive capacity, suggesting that neutrophils and oncostatin M may promote tumor progression in vivo.


Intercultural palliative care: do we need cultural competence?
Recognition of the importance of 'cultural competence' is now central to health care policy and to nurse education and training across the international spectrum. Detailed engagement with models of cultural competence is comparatively recent in palliative care nursing. This article presents the findings from a development project on elders and carers from 'minority ethnic' groups, funded by the Department of Health, to increase awareness of palliative care and to improve understanding of the needs of these groups of service users. The article describes the experiences of nurses involved in the delivery of palliative care who were interviewed in focus groups as a part of the project. It draws attention to the complicated relationships between cultural knowledge and practice and to the non-rational and visceral dimensions of intercultural care. These aspects of nursing are marginalised in current approaches to cultural competence, which emphasise the rational acquisition and application of cultural knowledge and skills by practitioners. It is suggested that recognition of these marginalised experiences can contribute to the development of new approaches to intercultural nursing that are also more attuned to the ethos and values of palliative care.


Results of the COMPARE trial of Constraint-induced or Multimodality Aphasia Therapy compared with usual care in chronic post-stroke aphasia.
While meta-analyses confirm treatment for chronic post-stroke aphasia is effective, a lack of comparative evidence for different interventions limits prescription accuracy. We investigated whether Constraint-Induced Aphasia Therapy Plus (CIAT-plus) and/or Multimodality Aphasia Therapy (M-MAT) provided greater therapeutic benefit compared with usual community care and were differentially effective according to baseline aphasia severity. We conducted a three-arm, multicentre, parallel group, open-label, blinded endpoint, phase III, randomised-controlled trial. We stratified eligible participants by baseline aphasia on the Western Aphasia Battery-Revised Aphasia Quotient (WAB-R-AQ). Groups of three participants were randomly assigned (1:1:1) to 30 hours of CIAT-Plus or M-MAT or to usual care (UC). Primary outcome was change in aphasia severity (WAB-R-AQ) from baseline to therapy completion analysed in the intention-to-treat population. Secondary outcomes included word retrieval, connected speech, functional communication, multimodal communication, quality of life and costs. We analysed 201 participants (70 in CIAT-Plus, 70 in M-MAT and 61 in UC). Aphasia severity was not significantly different between groups at postintervention: 1.05 points (95% CI -0.78 to 2.88; p=0.36) UC group vs CIAT-Plus; 1.06 points (95% CI -0.78 to 2.89; p=0.36) UC group vs M-MAT; 0.004 points (95% CI -1.76 to 1.77; p=1.00) CIAT-Plus vs M-MAT. Word retrieval, functional communication and communication-related quality of life were significantly improved following CIAT-Plus and M-MAT. Word retrieval benefits were maintained at 12-week follow-up. CIAT-Plus and M-MAT were effective for word retrieval, functional communication, and quality of life, while UC was not. Future studies should explore predictive characteristics of responders and impacts of maintenance doses. ACTRN 2615000618550.


A family process model of economic hardship and adjustment of early adolescent boys.
We propose a family process model that links economic stress in family life to prosocial and problematic adolescent adjustment. Employing a sample of 205 seventh-grade boys aged 12 to 14 years (M = 12.7) and living in intact families in the rural Midwest, the theoretical constructs in the model were measured using both trained observer and family member reports. In general, results were consistent with the proposed model. Objective economic conditions such as per capita income and unstable work were related to parents' emotional status and behaviors through their perceptions of increased economic pressures such as the inability to pay monthly bills. These pressures were associated with depression and demoralization for both parents, which was related to marital conflict and disruptions in skillful parenting. Disrupted parenting mediated the relations between the earlier steps in the stress process and adolescent adjustment. The emotions and behaviors of both mothers and fathers were almost equally affected by financial difficulties, and disruptions in each parent's child-rearing behaviors had adverse consequences for adolescent development.


Declining blood collection and utilization in the United States.
The Department of Health and Human Services National Blood Collection and Utilization Survey (NBCUS) has been conducted biennially since 1997. Data are used to estimate national blood collection and utilization. The 2013 Department of Health and Human Services NBCUS is a cross-sectional survey of all US blood collection centers and hospitals as listed in the 2012 American Hospital Association Annual Survey database that perform at least 100 inpatient surgical procedures annually. The study objective was to estimate, with 95% confidence intervals (CIs), the number of blood and blood components collected and transfused in the United States. In 2013, a total of 14,237,000 whole blood and apheresis red blood cell (RBC) units (95% CI, 13,639,000-14,835,000) were collected with 13,395,000 available for transfusion. Of these, 13,180,000 (95% CI, 12,389,000-13,972,000) whole blood and RBC units were transfused. This represented a 4.4% decline in the number of transfused units compared to 2011. Outdated (i.e., expired without being transfused) whole blood and RBC units declined by 17.3%. Apheresis (2,318,000; 95% CI, 2,154,000-2,482,000) and whole blood-derived platelet (PLT; 130,000; 95% CI, 23,000-237,000) distribution declined in 2013. Total PLT transfusions increased in 2013 (2,281,000) in comparison to 2011 (2,169,000). Total plasma units distributed (4,338,000) and transfused (3,624,000) declined. Both blood collection and utilization have declined, but the gap between collection and utilization is narrowing. As collections decline further and hospitals decrease transfusions and manage products more efficiently, the decline in surplus inventory may be a concern for disaster preparedness or other unexpected utilization needs.


Treatment of chromophobe adenmas with megavoltage tirradiation.
From 1956-1972, 62 previously untreated patients with chromophobe adenomas received high-dose (average equivalent dose: 5700rads in 6 weeks) megavoltage (4-4.8 MeV) irradiation at Stanford, 33 postoperatively and 29 as the only intended treatment. Initialtreatment failure rates were 18% and 41%, respectively; however, overall control was 85% and 90%, despite 2 uncontrolled "invasive" adenomas in each group. Nine of the 12 failures in the group treated by irradiation alone had cystic tumors, and 9 of the 12 "failed" in less than 3 months. Despite a considerably greater degree of abnormal vision initially in the postoperative irradiation group, improvement of vision with treatment in that group was 83% (19% returned to normal) compared to 46% (only 8% to normal) in the irradiation alone group. Based upon an evaluation of the extent of findings at diagnosis and our results, we recommend surgical decompression followed by 5000 rads in 5 weeks for patients with any one or more of the following findings: 1)more than minimal depression of peripheral visual fields; 2) corrected visual actuity of less than 20/30 in either eye; or 3)more than 1-cm suprasellar extension of tumor. We recommend irradiation alone, as specified, for smaller adenomas accompanied by less extensive or no visual abnormalities.


Texture Descriptors Based on Dijkstra's Algorithm for Medical Image Analysis.
In this paper we investigate a new approach for extracting features from a texture using Dijkstra's algorithm. The method maps images into graphs and gray level differences into transition costs. Texture is measured over the whole image comparing the costs found by Dijkstra's algorithm with the geometric distance of the pixels. In addition, we compare and combine our new strategy with a previous method for describing textures based on Dijkstra's algorithm. For each set of features, a support vector machine (SVM) is trained. The set of classifiers is then combined by weighted sum rule. Combining the proposed set of features with the well-known local binary patterns and local ternary patterns boosts performance. To assess the performance of our approach, we test it using six medical datasets representing different image classification problems. Tests demonstrate that our approach outperforms the performance of standard methods presented in the literature. All source code for the approaches tested in this paper will be available at: http://www.dei.unipd.it/node/2357.


Spatial Normalization of 18F-Flutemetamol PET Images Using an Adaptive Principal-Component Template
Though currently approved for visual assessment only, there is evidence to suggest that quantification of amyloid-β (Aβ) PET images may reduce interreader variability and aid in the monitoring of treatment effects in clinical trials. Quantification typically involves a regional atlas in standard space, requiring PET images to be spatially normalized. Different uptake patterns in Aβ-positive and Aβ-negative subjects, however, make spatial normalization challenging. In this study, we proposed a method to spatially normalize 18F-flutemetamol images using a synthetic template based on principal-component images to overcome these challenges. Methods: 18F-flutemetamol PET and corresponding MR images from a phase II trial (n = 70), including subjects ranging from Aβ-negative to Aβ-positive, were spatially normalized to standard space using an MR-driven registration method (SPM12). 18F-flutemetamol images were then intensity-normalized using the pons as a reference region. Principal-component images were calculated from the intensity-normalized images. A linear combination of the first 2 principal-component images was then used to model a synthetic template spanning the whole range from Aβ-negative to Aβ-positive. The synthetic template was then incorporated into our registration method, by which the optimal template was calculated as part of the registration process, providing a PET-only–driven registration method. Evaluation of the method was done in 2 steps. First, coregistered gray matter masks generated using SPM12 were spatially normalized using the PET- and MR-driven methods, respectively. The spatially normalized gray matter masks were then visually inspected and quantified. Second, to quantitatively compare the 2 registration methods, additional data from an ongoing study were spatially normalized using both methods, with correlation analysis done on the resulting cortical SUV ratios. Results: All scans were successfully spatially normalized using the proposed method with no manual adjustments performed. Both visual and quantitative comparison between the PET- and MR-driven methods showed high agreement in cortical regions. 18F-flutemetamol quantification showed strong agreement between the SUV ratios for the PET- and MR-driven methods (R2 = 0.996; pons reference region). Conclusion: The principal-component template registration method allows for robust and accurate registration of 18F-flutemetamol images to a standardized template space, without the need for an MR image.


Evolutionary patterns in prokaryotic genomes.
Prokaryotic genomics is shifting towards comparative approaches to unravel how and why genomes change over time. Both phylogenetic and population genetics approaches are required to dissect the relative roles of selection and drift under these conditions. Lineages evolve adaptively by selection of changes in extant genomes and the way this occurs is being explored from a systemic and evolutionary perspective to understand how mutations relate with gene repertoire changes and how both are contextualized in cellular networks. Through an increased appreciation of genome dynamics in given ecological contexts, a more detailed picture of the genetic basis of prokaryotic evolution is emerging.


The mammalian unfolded protein response.
In the endoplasmic reticulum (ER), secretory and transmembrane proteins fold into their native conformation and undergo posttranslational modifications important for their activity and structure. When protein folding in the ER is inhibited, signal transduction pathways, which increase the biosynthetic capacity and decrease the biosynthetic burden of the ER to maintain the homeostasis of this organelle, are activated. These pathways are called the unfolded protein response (UPR). In this review, we briefly summarize principles of protein folding and molecular chaperone function important for a mechanistic understanding of UPR-signaling events. We then discuss mechanisms of signal transduction employed by the UPR in mammals and our current understanding of the remodeling of cellular processes by the UPR. Finally, we summarize data that demonstrate that UPR signaling feeds into decision making in other processes previously thought to be unrelated to ER function, e.g., eukaryotic starvation responses and differentiation programs.


Combining simple blood tests to identify primary care patients with unexpected weight loss for cancer investigation: Clinical risk score development, internal validation, and net benefit analysis.
Unexpected weight loss (UWL) is a presenting feature of cancer in primary care. Existing research proposes simple combinations of clinical features (risk factors, symptoms, signs, and blood test data) that, when present, warrant cancer investigation. More complex combinations may modify cancer risk to sufficiently rule-out the need for investigation. We aimed to identify which clinical features can be used together to stratify patients with UWL based on their risk of cancer. We used data from 63,973 adults (age: mean 59 years, standard deviation 21 years; 42% male) to predict cancer in patients with UWL recorded in a large representative United Kingdom primary care electronic health record between January 1, 2000 and December 31, 2012. We derived 3 clinical prediction models using logistic regression and backwards stepwise covariate selection: Sm, symptoms-only model; STm, symptoms and tests model; Tm, tests-only model. Fifty imputations replaced missing data. Estimates of discrimination and calibration were derived using 10-fold internal cross-validation. Simple clinical risk scores are presented for models with the greatest clinical utility in decision curve analysis. The STm and Tm showed improved discrimination (area under the curve ≥ 0.91), calibration, and greater clinical utility than the Sm. The Tm was simplest including age-group, sex, albumin, alkaline phosphatase, liver enzymes, C-reactive protein, haemoglobin, platelets, and total white cell count. A Tm score of 5 balanced ruling-in (sensitivity 84.0%, positive likelihood ratio 5.36) and ruling-out (specificity 84.3%, negative likelihood ratio 0.19) further cancer investigation. A Tm score of 1 prioritised ruling-out (sensitivity 97.5%). At this threshold, 35 people presenting with UWL in primary care would be referred for investigation for each person with cancer referred, and 1,730 people would be spared referral for each person with cancer not referred. Study limitations include using a retrospective routinely collected dataset, a reliance on coding to identify UWL, and missing data for some predictors. Our findings suggest that combinations of simple blood test abnormalities could be used to identify patients with UWL who warrant referral for investigation, while people with combinations of normal results could be exempted from referral.


Direct comparative effectiveness and safety between non-vitamin K antagonist oral anticoagulants for stroke prevention in nonvalvular atrial fibrillation: a systematic review and meta-analysis of observational studies.
The non-vitamin K antagonist oral anticoagulants (NOACs) have been increasingly prescribed in clinical practice for stroke prevention in patients with nonvalvular atrial fibrillation (AF). Direct comparisons between NOACs in trials are lacking, leaving an important clinical decision-making gap. We aimed to perform a systematic review and meta-analysis to summarize the evidence of observational studies for direct comparative effectiveness and safety amongst NOACs in patients with AF. Conference proceedings and electronic databases including MEDLINE, CINAHL, EMBASE and PUBMED were systematically searched. We included observational studies directly comparing individual NOACs in patients with nonvalvular AF who were aged ≥ 18 years for stroke prevention. Primary outcome included effectiveness outcome (stroke or systemic embolism) and safety outcome (major bleeding). Data were extracted in duplicated by two reviewers independently. A random-effects meta-analysis was conducted to synthesize the data from included observational studies. We used the Grading of Recommendations Assessment, Development and Evaluation (GRADE) to rate the overall quality of evidence for each outcome. Fifteen studies were included for qualitative synthesis, twelve studies for meta-analyses. It was found that rivaroxaban and dabigatran were similar with regard to risk of stroke or systemic embolism (Hazard ratio [HR] = 1.00, 95% CI 0.91-1.10; evidence quality: low), but rivaroxaban was associated with higher risk of major bleeding (HR = 1.39, 95% CI 1.28-1.50; evidence quality: moderate). Compared with apixaban, a significantly higher risk of major bleeding was observed with rivaroxaban (HR = 1.71, 95% CI 1.51-1.94; evidence quality: low). Apixaban was associated with lower risk of major bleeding, in comparison with dabigatran (HR = 0.80, 95% CI 0.68-0.95; evidence quality: low). No differences in risk of stroke or systemic embolism was observed between rivaroxaban versus apixaban, and apixaban versus dabigatran. In this study, apixaban was found to have the most favorable safety profile amongst the three NOACs. No significant difference was observed in risk of stroke or systemic embolism between the NOACs. Such findings may provide some decision-making support for physicians regarding their choices amongst NOACs in patients with AF.Registration PROSPERO (identifier: CRD42016052908).


A comparison of the effects of rapid maxillary expansion and fan-type rapid maxillary expansion on dentofacial structures.
The aim of this study was to evaluate and compare the sagittal, transverse, and vertical effects of rapid maxillary expansion (RME) and fan-type RME on dentofacial structures. The study group consisted of 34 patients, 14 boys and 20 girls (average age 12.5 years), selected without considering their skeletal class and sex. The fan-type RME group comprised 17 subjects, who had an anterior constricted maxilla with a normal intermolar width. The RME group comprised 17 other subjects, who had a maxillary transverse discrepancy with a posterior crossbite. The records obtained for each patient included a lateral and a frontal cephalometric film, upper plaster models, and occlusal radiograph obtained before treatment (T1), after expansion (T2), and immediately after a three-month retention period (T3). The data obtained from the evaluation of the records before and after treatment, after treatment and after retention, and before treatment and after retention were compared using paired t-test. Further comparisons between the groups were made using Student's t-test. There was significantly greater expansion in the intercanine than in the intermolar width in the fan-type RME group as compared with the RME group. Downward and forward movement of the maxilla was observed in both groups. The upper incisors were tipped palatally in the RME group, but they were tipped labially in the fan-type RME group. There was significantly greater expansion in the nasal cavity and maxillary width in the RME group as opposed to the fan-type RME group.


Sex-dependent effects of nutrition on telomere dynamics in zebra finches (Taeniopygia guttata).
At a cellular level, oxidative stress is known to increase telomere attrition, and hence cellular senescence and risk of disease. It has been proposed that dietary micronutrients play an important role in telomere protection due to their antioxidant properties. We experimentally manipulated dietary micronutrients during early life in zebra finches (Taeniopygia guttata). We found no effects of micronutrient intake on telomere loss during chick growth. However, females given a diet high in micronutrients during sexual maturation showed reduced telomere loss; there was no such effect in males. These results suggest that micronutrients may influence rates of cellular senescence, but differences in micronutrient requirement and allocation strategies, probably linked to the development of sexual coloration, may underlie sex differences in response.


Electroencephalographic correlates of states of concentrative meditation.
Meditative techniques aim for and meditators report states of mental alertness and focus, concurrent with physical and emotional calm. We aimed to determine the electroencephalographic (EEG) correlates of five states of Buddhist concentrative meditation, particularly addressing a correlation with meditative level. We studied 12 meditators and 12 pair-matched meditation-naïve participants using high-resolution scalp-recorded EEG. To maximise reduction of EMG, data were pre-processed using independent component analysis and surface Laplacian transformed data. Two non-meditative and five meditative states were used: resting baseline, mind-wandering, absorptions 1, 2, 3, 4 and 5 (corresponding to four levels of absorption and an absorption with a different object of focus, otherwise equivalent to level 4; these five meditative states produce repeatable, distinctly different experiences for experienced meditators). The experimental protocol required participants to experience the states in the order listed above, followed immediately by the reverse. We then calculated EEG power in standard frequency bands from 1 to 80Hz. We observed decreases of central scalp beta (13-25Hz), and central low gamma (25-48Hz) power in meditators during deeper absorptions. In contrast, we identified increases in frontal midline and temporo-parietal theta power in meditators, again, during deeper absorptions. Alpha activity was increased over all meditative states, not depth-related. This study demonstrates that the subjective experiences of deepening meditation partially correspond to measures of EEG. Our results are in accord with prior studies on non-graded meditative states. These results are also consistent with increased theta correlating with tightness of focus, and reduced beta/gamma with the desynchronization associated with enhanced alertness.


Neuroendocrine and stress-related aspects of endometriosis.
Endometriosis is a model of a benign gynecologic disease associated with two major symptoms: pain and infertility. When becomes chronic, severe psychological and neuroendocrine changes may occur. The high levels of perceived stress caused by symptoms cause a neuroendocrine disequilibrium thus contributing to the progression of the disease. Elevated stress levels alter hormonal secretions, mood and behavior, sexual disorders and appetite custom. Inflammatory comorbidities may be associated with elevated stress in endometriotic patients (inflammatory bowel disease, fibromyalgia, chronic fatigue) and even autoimmune diseases (thyroid disease, systemic lupus erythematosus, multiple sclerosis). Neurogenic mechanisms are described in endometriotic lesions and they affect peripheral and central nervous system of these patients increasing pain sensitivity and stress reactivity. In conclusion, endometriosis is a disease which affects reproductive and neuroendocrine functions with a great impact on women's health and quality of life.


Hydroxylation of p-nitrophenol by rabbit ethanol-inducible cytochrome P-450 isozyme 3a.
The hydroxylation of p-nitrophenol to 4-nitrocatechol was investigated using rabbit hepatic microsomes and six purified isozymes of cytochrome P-450. The microsomal activity was maximal at pH 6.8 and at 100 microM p-nitrophenol. At higher substrate concentrations inhibition was observed. At pH 6.8 and 100 microM p-nitrophenol, isozyme 3a exhibited the highest activity of the purified isozymes: 3.4-fold more active than isozyme 6, and 8-fold more active than isozymes 2 and 4. The isozyme 3a-catalyzed hydroxylation reaction was stimulated 2.4-fold by the addition of a 4:1 ratio of cytochrome b5/P-450. At optimal concentrations of cytochrome b5, isozyme 3a was 8- to 9-fold more active than isozymes 2 and 6 and 20-fold more active than isozyme 4. Under the same conditions, isozyme 3a-catalyzed butanol oxidation was inhibited 40%. Antibodies to isozyme 3a inhibited greater than 95% of the p-nitrophenol hydroxylase activity of microsomes from untreated or from ethanol- or acetone-treated rabbits. The microsomal hydroxylase activity was linearly correlated with the microsomal concentration of isozyme 3a (correlation coefficient of 0.94) and had an intercept near zero. The results from reconstitution, antibody inhibition, and correlation experiments indicate that isozyme 3a is the principal catalyst of rabbit microsomal p-nitrophenol hydroxylation. The ability of the ethanol-inducible isozyme to catalyze catechol formation may be important in the ethanol-enhanced toxicity of aromatic compounds such as benzene.


Longitudinal Lung Function Decrease in Subjects with Spontaneous Healed Pulmonary Tuberculosis.
We compared the longitudinal course of post-bronchodilator Forced Expiratory Volume in 1 second (pFEV1) over a 10-year period in subjects with spontaneous healed pulmonary tuberculosis (SHPTB) with that in normal subjects. We prospectively investigated 339 subjects with SHPTB and 3211 normal subjects. pFEV1 values measured biannually over 10 years were analyzed using mixed effects model. At baseline, there were no differences in gender, smoking amount, and mean height, except mean age (50.0 ± 8.1 VS. 48.1 ± 7.3, P< 0.001) between the SHPTB and normal group. 52% of the 339 participants with SHPTB and 56% of the 3211 normal participants participated till the end of study. According to the final model, the SHPTB group showed significantly larger decrease in the average pFEV1 over the time than the normal group (P< 0.001) adjusted for gender, age, height, smoking pack years, and time effects. Especially, the interaction effect between time and group was statistically significant (P = 0.036). The average lung function in terms of pFEV1 decreases faster in subjects with SHPTB than in normal individuals over time.


Process Evaluation of Cancer Prevention Media Campaigns in Appalachian Ohio.
The purpose of this study is to provide process data from campaigns (2009-2010) to improve colorectal cancer (CRC; intervention) screening and fruit and vegetable (F&V; comparison) consumption in 12 Appalachian Ohio counties. County-specific campaigns included one billboard, posters, and articles for local newspapers. Participants in CRC screening counties who reported seeing CRC screening billboards had greater intention to talk to a doctor/nurse about screening in the next 6 months (odds ratio [OR] = 2.92, 95% confidence interval [CI; 1.71, 4.99]) and had twice the odds of talking to a doctor/nurse about screening in the past year (OR = 2.15, 95% CI [1.29, 3.60]) compared to those who did not see the billboards. Participants in F&V counties who reported seeing F&V billboards had twice the odds (OR = 2.27, 95% CI [1.35, 3.84]) of talking to a doctor/nurse in the past year about F&Vs compared to those who did not see the billboards. Participants who reported campaign exposure lived closer to the billboards compared to those who did not report campaign exposure (mean distance in miles from home to billboard: 8.8 vs. 10.9; p < .01). Most participants reported campaign messages were clear and important. Results suggest that partnering with community members to develop campaign materials is important to ensure cultural appropriateness and that exposure to the intervention components may affect health-related outcomes.


Dried plum polyphenols inhibit osteoclastogenesis by downregulating NFATc1 and inflammatory mediators.
Dried plums and their polyphenols have been shown to suppress bone resorption by downregulating receptor activator NF-kappaB ligand (RANKL). Due to the anti-inflammatory and antioxidant properties of these compounds, this study was designed to investigate whether dried plum polyphenols exert additional, more direct effects on osteoclasts and their precursors. RAW 264.7 macrophages were used as a model to study osteoclast precursors and osteoclast differentiation and activity. Under inflammatory conditions induced by lipopolysaccharide (LPS), polyphenols extracted from dried plum (10, 20, and 30 microg/mL) downregulated osteoclast precursor cyclooxygenase expression and nitric oxide (NO) by inhibiting inducible NO synthase. NO and tumor necrosis factor (TNF)-alpha were also suppressed in the presence of RANKL during osteoclastogenesis by the polyphenols. Increased TNF-alpha production in response to oxidative stress, but not LPS, was decreased over time. As expected, LPS and H2O2 significantly increased the number of tartrate-resistant acid phosphatase-positive cells by 127% and 30%, respectively. Dried plum polyphenols decreased osteoclast differentiation under normal as well as inflammatory and oxidative stress conditions, coincident with the suppression of the transcription factor, nuclear factor for activated T cells (NFATcl). These inhibitory effects on osteoclastogenesis were confirmed in primary bone marrow cultures. Resorption pit formation was decreased to a similar extent as osteoclast differentiation, suggesting that dried plum polyphenols primarily affect osteoclast differentiation as opposed to activity. Our data demonstrate that dried plum polyphenols directly inhibit osteoclastogenesis, leading to a decrease in osteoclast activity, by downregulating NFATc1 and inflammatory mediators.


Cigarette smoke promotes dendritic cell accumulation in COPD; a Lung Tissue Research Consortium study.
Abnormal immune responses are believed to be highly relevant in the pathogenesis of chronic obstructive pulmonary disease (COPD). Dendritic cells provide a critical checkpoint for immunity by their capacity to both induce and suppress immunity. Although evident that cigarette smoke, the primary cause of COPD, significantly influences dendritic cell functions, little is known about the roles of dendritic cells in the pathogenesis of COPD. The extent of dendritic cell infiltration in COPD tissue specimens was determined using immunohistochemical localization of CD83+ cells (marker of matured myeloid dendritic cells), and CD1a+ cells (Langerhans cells). The extent of tissue infiltration with Langerhans cells was also determined by the relative expression of the CD207 gene in COPD versus control tissues. To determine mechanisms by which dendritic cells accumulate in COPD, complimentary studies were conducted using monocyte-derived human dendritic cells exposed to cigarette smoke extract (CSE), and dendritic cells extracted from mice chronically exposed to cigarette smoke. In human COPD lung tissue, we detected a significant increase in the total number of CD83+ cells, and significantly higher amounts of CD207 mRNA when compared with control tissue. Human monocyte-derived dendritic cells exposed to CSE (0.1-2%) exhibited enhanced survival in vitro when compared with control dendritic cells. Murine dendritic cells extracted from mice exposed to cigarette smoke for 4 weeks, also demonstrated enhanced survival compared to dendritic cells extracted from control mice. Acute exposure of human dendritic cells to CSE induced the cellular pro-survival proteins heme-oxygenase-1 (HO-1), and B cell lymphoma leukemia-x(L) (Bcl-xL), predominantly through oxidative stress. Although activated human dendritic cells conditioned with CSE expressed diminished migratory CCR7 expression, their migration towards the CCR7 ligand CCL21 was not impaired. These data indicate that COPD is associated with increased numbers of cells bearing markers associated with Langerhans cells and mature dendritic cells, and that cigarette smoke promotes survival signals and augments survival of dendritic cells. Although CSE suppressed dendritic cell CCR7 expression, migration towards a CCR7 ligand was not diminished, suggesting that reduced CCR7-dependent migration is unlikely to be an important mechanism for dendritic cell retention in the lungs of smokers with COPD.


Metabolic abnormalities in an early psychosis service: a retrospective, naturalistic cross-sectional study.
There is an increasing recognition of the impact of weight gain on the development of metabolic abnormalities in young people receiving atypical antipsychotic drugs for first-episode psychosis. This study examined the prevalence of such abnormalities in a specialist early-intervention community mental health team. A retrospective case record audit of 85 patients 16-27 years old attending the Early Psychosis Service between October 2006 and June 2008, who had at least one metabolic measure defined as: weight, body mass index (BMI), waist circumference, blood pressure, and fasting blood glucose and lipids. Metabolic syndrome identified by the International Diabetes Federation (IDF) criteria. Fifty-five percent of males and 42% of females were overweight or obese at a median treatment duration of 8 months. Duration of antipsychotic therapy was associated with higher BMI (r = 0.28, P < 0.01). More than 40% of the total sample had high waist circumference. Of the 64 subjects with complete metabolic data, eight (12.5%) met full IDF criteria for metabolic syndrome, and another 21 (32.8%) had either increased waist with one metabolic abnormality or normal waist and two metabolic abnormalities. Over a third of young patients being treated for their first episode of psychosis either had metabolic syndrome or showed metabolic abnormalities. Treatment duration related to higher BMI and greater prevalence of metabolic syndrome. Detection of metabolic complications after treatment instigation in patients with first-episode psychosis will permit early intervention with lifestyle or drug interventions in those at risk of significant physical health morbidity.


A Practical Application of Solid-phase Extraction Using a Syringe Filled with Sorbent for the Determination of Lead and Cadmium in Water.
In this study, at first the synthesis of 3-chloro-2-hydroxypropyl methacrylate-ethylene glycole dimethacrylate co-polymer beads and its modification with tris(2-aminoethyl) amine is described. Characterization of the polymer was done by FTIR and SEM. The functional co-polymer was filled in a disposable pipet tip and tightly connected to a 50-mL syringe for the separation and the enrichment of lead and cadmium prior to their determination by flame atomic absorption spectrometry. The sample and then the eluate were subsequently drawn and discharged to retain and desorb lead and cadmium by means of the syringe, respectively. Both analytes were quantitatively retained at pH 4 and eluted using 3.0 mol L-1 of HNO3 at flow rates of approximately 10 mL min-1. Under the optimum conditions, the enrichment factors of up to 50-fold both elements could be obtained by drawing and discharging 250 mL (5 × 50 mL) of the sample, and then 5 mL of the eluent. The recoveries were >90%. The limits of detection (3σ; N = 10 of blank) for Pb and Cd were 0.0034 and 0.0016 mg L-1 for a 50-fold enrichment, respectively. The analyte concentrations in a certified waste water reference agreed within the certified values in the 95% confidence range.


Agranulocytosis in Bangkok, Thailand: a predominantly drug-induced disease with an unusually low incidence. Aplastic Anemia Study Group.
Agranulocytosis, a syndrome characterized by a marked reduction in circulating granulocytes, is strongly associated with medical drug use in Europe and the United States. Unregulated use of common pharmaceutical agents in developing countries has been suspected of causing large numbers of cases of agranulocytosis and deaths, especially among children. To elucidate the incidence and etiology of agranulocytosis in Thailand, a population-based case-control study of symptomatic agranulocytosis that resulted in hospital admission was conducted in Bangkok from 1990 to 1994. An attempt was also made to study the disease in Khonkaen (in northeastern Thailand) and Songkla (in southern Thailand), but there were insufficient cases in the latter regions, and the analysis was confined to subjects from Bangkok. In that region, the overall incidence of agranulocytosis was 0.8 per million per year; there were no deaths. As expected, the incidence was higher in females (0.9 per million), and it increased with age (4.3 per million beyond age 60). Among 25 cases and 529 controls the relative risk estimate for a combined category of all suspect drugs was 9.2 (95% confidence interval = 3.9-21), and the proportion of cases that could be attributed to drug use was 68%. For individual drugs and drug classes the data were sparse; within these limitations, the strongest association appeared to be with antithyroid drugs. One case and three controls were exposed to dipyrone, a drug known to cause agranulocytosis; with such scanty data the risk could not be evaluated. Exposure to pesticides or solvents was not associated with an increased risk. This is the first formal epidemiologic study of agranulocytosis in a developing country. As in the West, most cases are attributable to medical drug use. However, the incidence of agranulocytosis in Bangkok, and apparently, in Thailand as a whole, is unusually low, and the disease does not pose a public health risk.


Spatial Lifecourse Epidemiology Reporting Standards (ISLE-ReSt) statement.
Spatial lifecourse epidemiology is an interdisciplinary field that utilizes advanced spatial, location-based, and artificial intelligence technologies to investigate the long-term effects of environmental, behavioural, psychosocial, and biological factors on health-related states and events and the underlying mechanisms. With the growing number of studies reporting findings from this field and the critical need for public health and policy decisions to be based on the strongest science possible, transparency and clarity in reporting in spatial lifecourse epidemiologic studies is essential. A task force supported by the International Initiative on Spatial Lifecourse Epidemiology (ISLE) identified a need for guidance in this area and developed a Spatial Lifecourse Epidemiology Reporting Standards (ISLE-ReSt) Statement. The aim is to provide a checklist of recommendations to improve and make more consistent reporting of spatial lifecourse epidemiologic studies. The STrengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement for cohort studies was identified as an appropriate starting point to provide initial items to consider for inclusion. Reporting standards for spatial data and methods were then integrated to form a single comprehensive checklist of reporting recommendations. The strength of our approach has been our international and multidisciplinary team of content experts and contributors who represent a wide range of relevant scientific conventions, and our adherence to international norms for the development of reporting guidelines. As spatial, location-based, and artificial intelligence technologies used in spatial lifecourse epidemiology continue to evolve at a rapid pace, it will be necessary to revisit and adapt the ISLE-ReSt at least every 2-3 years from its release.


Botulinum toxin for myofascial pain syndromes in adults.
This is an updated version of the original Cochrane review published in Issue 4, 2012. Myofascial pain syndrome (MPS) is a regional muscular pain syndrome characterised by the presence of trigger points, which are painful points in one or more muscles. The pain can be felt at the site where the trigger point is located or it can be felt away from that place when the muscle is pressed (referred pain). Botulinum toxin is a protein produced by the bacterium Clostridium botulinum and is a potent neurotoxin that eventually inhibits muscle contractions. It is capable of selectively weakening painful muscles and interrupting the pain cycle. To assess the effectiveness and safety of botulinum toxin A (BTXA) in the treatment of myofascial pain syndrome (MPS), excluding MPS in neck and head muscles. This is an updated version of the original Cochrane review published in Issue 4, 2012. The search strategy for the update was the same as in the original review and we searched CENTRAL in The Cochrane Library (2013, Issue 11 of 12), MEDLINE (Ovid) (2012 to 29 November 2013) and EMBASE (Ovid) (2012 to 27 November 2013). The search strategy was composed of terms for myofascial pain and botulinum toxin. For the original review, we also searched the Cochrane Pain, Palliative and Supportive Care (PaPaS) Review Group Specialised Register until December 2011, PubMed (from 1966 to 2011) and LILACS (from 1982 to 2011). There was no language restriction. We included randomised controlled trials (RCTs) involving botulinum toxin for treating participants with MPS. We excluded studies with MPS of the neck and head from this review as they have already been assessed in existing systematic reviews. We considered a diagnosis of MPS to be based on the identification of trigger points in the taut band through palpation of sensitive nodules, local twitch response and specific patterns of referred pain associated with each trigger point. Two review authors independently screened identified studies, extracted data, assessed trial quality and analysed results using the Cochrane PaPaS Review Group criteria. Four studies with a total of 233 participants, comparing BTXA with placebo, met the inclusion criteria. In one study with 145 participants, significant improvement rates of pain intensity scores and duration of daily pain were demonstrated when comparing BTXA with placebo. The three other studies showed that there was no statistically significant difference between BTXA and placebo in pain intensity. Since the first publication of this review, no new studies were found. There is inconclusive evidence to support the use of botulinum toxin in the treatment of MPS based on data from four studies with a total of 233 participants, which we considered were of sufficient quality to be included in this review. Meta-analyses were not possible due to the heterogeneity between studies. We suggest that in future studies the same methodology to assess pain, a standardised dose of treatment, follow-up of at least four months (to observe the maximum and minimum curve of the drug effect) and appropriate data presentation should be used. More high-quality RCTs of botulinum toxin for treating MPS need to be conducted before firm conclusions on its effectiveness and safety can be drawn.