Antidepressants and suicide attempts in children.
Recent data showing possible increased risk for suicidal behavior among children and adolescents treated with selective serotonin reuptake inhibitors (SSRIs) and serotonin-norepinephrine reuptake inhibitors (SNRIs) antidepressants have created significant concern among patients, families, and providers, including concerns about the risk of individual antidepressants. This study was designed to compare the risk for medically treated suicide attempts among new users of sertraline, paroxetine, citalopram, escitalopram, and venlafaxine to risk for new users of fluoxetine. A retrospective cohort study included 36,842 children aged 6 to 18 years enrolled in Tennessee Medicaid between 1995 and 2006 who were new users of 1 of the antidepressant medications of interest (defined as filling no prescriptions for antidepressants in the preceding 365 days). Medically treated suicide attempts were identified from Medicaid files and vital records and confirmed with medical record review. Four hundred nineteen cohort members had a medically treated suicide attempt with explicit or inferred attempt to die confirmed through medical record review, including 4 who completed suicide. The rate of confirmed suicide attempts for the study drugs ranged from 24.0 per 1000 person-years to 29.1 per 1000 person-years. The adjusted rate of suicide attempts did not differ significantly among current users of SSRI and SNRI antidepressants compared with current users of fluoxetine. Users of multiple antidepressants concomitantly had increased risk for suicide attempt. In this population-based study of children recently initiating an antidepressant, there was no evidence that risk of suicide attempts differed for commonly prescribed SSRI and SNRI antidepressants.


Recent molecular discoveries in angiogenesis and antiangiogenic therapies in cancer.
Four decades ago, angiogenesis was recognized as a therapeutic target for blocking cancer growth. Because of its importance, VEGF has been at the center stage of antiangiogenic therapy. Now, several years after FDA approval of an anti-VEGF antibody as the first antiangiogenic agent, many patients with cancer and ocular neovascularization have benefited from VEGF-targeted therapy; however, this anticancer strategy is challenged by insufficient efficacy, intrinsic refractoriness, and resistance. Here, we examine recent discoveries of new mechanisms underlying angiogenesis, discuss successes and challenges of current antiangiogenic therapy, and highlight emerging antiangiogenic paradigms.


Targeting G protein-coupled receptors for the treatment of chronic pain in the digestive system.
Chronic pain is a hallmark of functional disorders, inflammatory diseases and cancer of the digestive system. The mechanisms that initiate and sustain chronic pain are incompletely understood, and available therapies are inadequate. This review highlights recent advances in the structure and function of pronociceptive and antinociceptive G protein-coupled receptors (GPCRs) that provide insights into the mechanisms and treatment of chronic pain. This knowledge, derived from studies of somatic pain, can guide research into visceral pain. Mediators from injured tissues transiently activate GPCRs at the plasma membrane of neurons, leading to sensitisation of ion channels and acute hyperexcitability and nociception. Sustained agonist release evokes GPCR redistribution to endosomes, where persistent signalling regulates activity of channels and genes that control chronic hyperexcitability and nociception. Endosomally targeted GPCR antagonists provide superior pain relief in preclinical models. Biased agonists stabilise GPCR conformations that favour signalling of beneficial actions at the expense of detrimental side effects. Biased agonists of µ-opioid receptors (MOPrs) can provide analgesia without addiction, respiratory depression and constipation. Opioids that preferentially bind to MOPrs in the acidic microenvironment of diseased tissues produce analgesia without side effects. Allosteric modulators of GPCRs fine-tune actions of endogenous ligands, offering the prospect of refined pain control. GPCR dimers might function as distinct therapeutic targets for nociception. The discovery that GPCRs that control itch also mediate irritant sensation in the colon has revealed new targets. A deeper understanding of GPCR structure and function in different microenvironments offers the potential of developing superior treatments for GI pain.


The emergence of new psychoactive substance (NPS) benzodiazepines: A review.
The market for new psychoactive substances has increased markedly in recent years and there is now a steady stream of compounds appearing every year. Benzodiazepines consist of only a fraction of the total number of these compounds but their use and misuse has rapidly increased. Some of these benzodiazepines have only been patented, some of them have not been previously synthesised, and the majority have never undergone clinical trials or tests. Despite their structural and chemical similarity, large differences exist between the benzodiazepines in their pharmacokinetic parameters and metabolic pathways and so they are not easily comparable. As benzodiazepines have been clinically used since the 1960s, many analytical methods exist to quantify them in a variety of biological matrices and it is expected that these methods would also be suitable for the detection of benzodiazepines that are novel psychoactive substances. Illicitly obtained benzodiazepines have been found to contain a wide range of compounds such as opiates which presents a problem since the use of them in conjunction with each other can lead to respiratory depression and death. This review collates the available information on these benzodiazepines and provides a starting point for the further investigation of their pharmacokinetics which is clearly required.


Outcomes after liver transplantation in young infants.
Liver transplantation in infants younger than 90 days is increasingly common. These infants typically arrive for transplantation in fragile medical condition. It is commonly assumed that they may experience high complication rates, difficult postoperative courses, and poor graft and patient survival. We sought to understand whether graft and patient survival rates in these young infants were lower than in older children, these patients experienced more complications than older children, and health care resource utilization was higher in this population. Data queried from the Study of Pediatric Liver Transplantation (SPLIT) database were limited to infants ages 0 to 90 days who had received their primary liver transplant between February 1996 and May 2004. Patients older than 90 days registered in the SPLIT database were used for comparison. Thirty-eight patients, ages 0 to 90 days, were included in the analysis. Their severity of illness was reflected by a median calculated Pediatric Endstage Liver Disease score of 34.8 at transplant. A majority (89.5%) of infants received cadaveric liver grafts, of which 47% were reduced organs. The infants experienced prolonged hospitalizations, spending an average of 50.9 +/- 7.6 days in the hospital after transplant. Long stays in the intensive care unit (average 22.1 +/- 1.5 days) and need for mechanical ventilation (average 16.2 +/- 2.7 days) also occurred. Length of hospitalization, intensive care, and mechanical ventilation were significantly higher than in older children (P < 0.0001). The reoperation rate (60.5%) was high and significantly greater than in older children (P = 0.007), with 10 patients (26.3%) needing 3 or more early reoperations. Reoperations occurred for bleeding, wound complications, biliary complications, and sepsis. There was no difference in vascular or biliary tract complications compared with older children. Bacterial infections were also common (52.6%) and were seen with greater frequency than in older children (P < 0.04). This infant cohort had an overall graft survival of 76.1% and overall patient survival of 87.8% at 1 year, with median follow-up of 12.5 months (range 0.6-84.0 months). Graft and patient survival in infants younger than 90 days was similar to that in older infants and children (P = NS). Young infants experience graft and patient survival similar to that in older cohorts of liver transplant recipients. Posttransplant complication rates, including the reoperation rate, were higher in this younger group, and the duration of hospitalization and intensive support were significantly longer. Future studies to better examine the factors, including age, that may contribute to the need for reoperation in children are warranted. Recognition and further analysis of the cost of care in this age group is also needed.


Dynamic covalent chemistry.
Dynamic covalent chemistry relates to chemical reactions carried out reversibly under conditions of equilibrium control. The reversible nature of the reactions introduces the prospects of "error checking" and "proof-reading" into synthetic processes where dynamic covalent chemistry operates. Since the formation of products occurs under thermodynamic control, product distributions depend only on the relative stabilities of the final products. In kinetically controlled reactions, however, it is the free energy differences between the transition states leading to the products that determines their relative proportions. Supramolecular chemistry has had a huge impact on synthesis at two levels: one is noncovalent synthesis, or strict self-assembly, and the other is supramolecular assistance to molecular synthesis, also referred to as self-assembly followed by covalent modification. Noncovalent synthesis has given us access to finite supermolecules and infinite supramolecular arrays. Supramolecular assistance to covalent synthesis has been exploited in the construction of more-complex systems, such as interlocked molecular compounds (for example, catenanes and rotaxanes) as well as container molecules (molecular capsules). The appealing prospect of also synthesizing these types of compounds with complex molecular architectures using reversible covalent bond forming chemistry has led to the development of dynamic covalent chemistry. Historically, dynamic covalent chemistry has played a central role in the development of conformational analysis by opening up the possibility to be able to equilibrate configurational isomers, sometimes with base (for example, esters) and sometimes with acid (for example, acetals). These stereochemical "balancing acts" revealed another major advantage that dynamic covalent chemistry offers the chemist, which is not so easily accessible in the kinetically controlled regime: the ability to re-adjust the product distribution of a reaction, even once the initial products have been formed, by changing the reaction's environment (for example, concentration, temperature, presence or absence of a template). This highly transparent, yet tremendously subtle, characteristic of dynamic covalent chemistry has led to key discoveries in polymer chemistry. In this review, some recent examples where dynamic covalent chemistry has been demonstrated are shown to emphasise the basic concepts of this area of science.


Oxidative and electrophilic stresses activate Nrf2 through inhibition of ubiquitination activity of Keap1.
The Keap1-Nrf2 system is the major regulatory pathway of cytoprotective gene expression against oxidative and/or electrophilic stresses. Keap1 acts as a stress sensor protein in this system. While Keap1 constitutively suppresses Nrf2 activity under unstressed conditions, oxidants or electrophiles provoke the repression of Keap1 activity, inducing the Nrf2 activation. However, the precise molecular mechanisms behind the liberation of Nrf2 from Keap1 repression in the presence of stress remain to be elucidated. We hypothesized that oxidative and electrophilic stresses induce the nuclear accumulation of Nrf2 by affecting the Keap1-mediated rapid turnover of Nrf2, since such accumulation was diminished by the protein synthesis inhibitor cycloheximide. While both the Cys273 and Cys288 residues of Keap1 are required for suppressing Nrf2 nuclear accumulation, treatment of cells with electrophiles or mutation of these cysteine residues to alanine did not affect the association of Keap1 with Nrf2 either in vivo or in vitro. Rather, these treatments impaired the Keap1-mediated proteasomal degradation of Nrf2. These results support the contention that Nrf2 protein synthesized de novo after exposure to stress accumulates in the nucleus by bypassing the Keap1 gate and that the sensory mechanism of oxidative and electrophilic stresses is closely linked to the degradation mechanism of Nrf2.


Combinatorial recognition of a complex telomere repeat sequence by the Candida parapsilosis Cdc13AB heterodimer.
The telomere repeat units of Candida species are substantially longer and more complex than those in other organisms, raising interesting questions concerning the recognition mechanisms of telomere-binding proteins. Herein we characterized the properties of Candida parapsilosis Cdc13A and Cdc13B, two paralogs that are responsible for binding and protecting the telomere G-strand tails. We found that Cdc13A and Cdc13B can each form complexes with itself and a heterodimeric complex with each other. However, only the heterodimer exhibits high-affinity and sequence-specific binding to the telomere G-tail. EMSA and crosslinking analysis revealed a combinatorial mechanism of DNA recognition, which entails the A and B subunit making contacts to the 3' and 5' region of the repeat unit. While both the DBD and OB4 domain of Cdc13A can bind to the equivalent domain in Cdc13B, only the OB4 complex behaves as a stable heterodimer. The unstable Cdc13AB(DBD) complex binds G-strand with greatly reduced affinity but the same sequence specificity. Thus the OB4 domains evidently contribute to binding by promoting dimerization of the DBDs. Our investigation reveals a rare example of combinatorial recognition of single-stranded DNA and offers insights into the co-evolution of telomere DNA and cognate binding proteins.


Factors Associated with the Timeliness of Electronic Nursing Documentation.
To investigate the factors associated with the timeliness of electronic nursing documentation using the entry time on the Electronic Medical Record (EMR) system. As a retrospective study, data were extracted from January 1 to February 28, 2014 from a hospital EMR system and a nurses' personnel information system. The timeliness of instances of nursing documentation was categorized into 'timely' or 'untimely' according to whether the entry time was time-stamped within the working hours during each day, evening, or night shift. Factors associated with the timeliness of the electronic nursing documentation were included in the logistic regression models as nurse- and patient-associated factors. Among 1,700,247 instances of electronic nursing documentation, 79.3% (n = 1,347,711) were completed within the working hours. Years of nursing experience, nursing shift, days of the week, patients' age, and medical department had a statistically significant associated with the timeliness of nursing records. Nurses with experience of more than 1 year entered nursing records over 2 times more during their working hours than did less experienced nurses. During the evening and night shifts, nurses were 1.49 times and 9.19 times more likely to enter nursing documents in a timely manner, respectively, as compared to those in the day shift. Nursing documentation was typically completed outside of working hours when a nurse had little experience, worked during the day shift or weekdays, and when tasks were unpredictable. This shows that new nurses need support to familiarize them with various tasks and the overall workflow.


Ligand-dependent interaction of estrogen receptor-alpha with members of the forkhead transcription factor family.
Estrogen acting through the estrogen receptor (ER) is able to regulate cell growth and differentiation of a variety of normal tissues and hormone-responsive tumors. Ligand-activated ER binds DNA and transactivates the promoters of estrogen target genes. In addition, ligand-activated ER can interact with other factors to alter the physiology and growth of cells. Using a yeast two-hybrid screen, we have identified an interaction between ER alpha and the proapoptotic forkhead transcription factor FKHR. The ER alpha-FKHR interaction depends on beta-estradiol and is reduced significantly in the absence of hormone or the presence of Tamoxifen. A glutathione S-transferase pull-down assay was used to confirm the interaction and localized two interaction sites, one in the forkhead domain and a second in the carboxyl terminus. The FKHR interaction was specific to ER alpha and was not detected with other ligand-activated steroid receptors. The related family members, FKHRL1 and AFX, also bound to ER alpha in the presence of beta-estradiol. FKHR augmented ER alpha transactivation through an estrogen response element. Conversely, ER alpha repressed FKHR-mediated transactivation through an insulin response sequence, and cell cycle arrest induced by FKHRL1 in MCF7 cells was abrogated by estradiol. These results suggest a novel mechanism of estrogen action that involves regulation of the proapoptotic forkhead transcription factors.


Comparison of diagnostic accuracy of combined assessment using adenosine stress computed tomography perfusion + computed tomography angiography with transluminal attenuation gradient + computed tomography angiography against invasive fractional flow reserve.
The goal of this study was to compare the diagnostic accuracy of combined computed tomography perfusion (CTP) + computed tomography angiography (CTA), transluminal attenuation gradient by 320-detector row computed tomography (TAG320) + CTA, and CTP + TAG320 + CTA (multidetector computed tomography-integrated protocol [MDCT-IP]) assessment in predicting significant fractional flow reserve (FFR). CTA has limited specificity for predicting functionally significant stenoses. Novel CT techniques, including adenosine stress CTP and TAG320, may improve the diagnostic accuracy of CTA. CTA, CTP, and TAG320 were assessed using 320-detector row MDCT. Patients who underwent CTA, CTP, and FFR assessment on invasive coronary angiography were included. CTP was assessed using the visual perfusion assessment. TAG320 was defined as the linear regression coefficient between luminal attenuation and axial distance. A TAG320 cutoff value of -15.1 HU/10 mm as previously described was defined as significant. Functionally significant coronary stenosis was defined as FFR ≤0.8. The cohort included 75 patients (age 64.1 ± 10.8 years, 52 men) and 44 (35%) FFR-significant vessels. In 127 vessels, CTA predicted FFR-significant stenosis with 89% sensitivity and 65% specificity compared with MDCT-IP, which showed 88% sensitivity and 83% specificity. In 97 vessels in which the results of all techniques were available, TAG320 + CTA (area under the curve [AUC] = 0.844) and CTP + CTA (AUC = 0.845) had comparable per-vessel diagnostic accuracy (p = 0.98). The diagnostic accuracy of MDCT-IP (AUC = 0.91) was superior to TAG320 + CTA or CTP + CTA (p = 0.01). In vessels without significant calcification or artefact, TAG320 + CTA and CTP + CTA provide comparable diagnostic accuracy for functional assessment of coronary artery stenosis. MDCT-IP may provide the best diagnostic accuracy for functional assessment of coronary artery stenosis.


Effects of euglycaemic and hypoglycaemic hyperinsulinaemia on sympathetic and parasympathetic regulation of haemodynamics in healthy subjects.
The effects of hypoglycaemia during hyperinsulinaemia, occurring under various pathophysiological conditions, on the cardiovascular regulatory system and vasculature are largely unknown. The aim of the present study was to investigate regulatory and haemodynamic responses to acute hyperinsulinaemia and consequent hypoglycaemia in 18 healthy subjects. Blood sampling and 5 min ECG and blood pressure recordings were performed at baseline and during the euglycaemic and hypoglycaemic phases of a hyperinsulinaemic clamp. Heart rate variability (HRV) and blood pressure variability (BPV) were assessed by using power spectral analysis, and baroreflex sensitivity (BRS) was assessed using the cross-spectral method. Stroke volume was assessed from the non-invasive blood pressure signal by the arterial pulse contour method. Euglycaemic hyperinsulinaemia did not change plasma catecholamine concentrations, HRV, BPV, BRS, heart rate, blood pressure, stroke volume, cardiac output or peripheral resistance. However, hyperinsulinaemic hypoglycaemia resulted in an 11.7-fold increase in the plasma adrenaline concentration (from 0.19+/-0.03 to 1.68+/-0.32 nmol/l; P <0.001), and a modest 1.3-fold increase in the plasma noradrenaline concentration (from 1.74+/-0.22 to 2.02+/-0.19 nmol/l; P <0.05) compared with baseline. Furthermore, we observed significant decreases in diastolic blood pressure (from 68+/-3 to 60+/-3 mmHg; P <0.05) and peripheral resistance (from 24.1+/-1.2 to 18.5+/-1.1 mmHg.min(-1) x l(-1); P <0.01). Stroke volume and cardiac output increased markedly from the euglycaemic to the hypoglycaemic period only ( P <0.01 for both). Hypoglycaemia did not influence HRV, BPV or BRS. Our findings indicate that hyperinsulinaemic hypoglycaemia is characterized by a significant increase in the plasma adrenaline concentration and by decreases in peripheral resistance and blood pressure. Counter-regulation during hyperinsulinaemic hypoglycaemia involves selective adrenomedullary sympathetic activation, and does not influence cardiac parasympathetic regulation or baroreflex control of heart rate.


A Phenomenological Analysis of the Spiritual Support Services in State Hospitals: The Case of Turkey.
The purpose of this research is to examine the core and nature of spiritual support services experienced by Turkey's pioneer spiritual support specialists (SSSs). A phenomenological approach was used in this qualitative research in which six males, two females, and eight SSSs participated. It has been seen that as a result of face-to-face interviews with participants within the framework of the semi-structured interview protocol, the spiritual support service has been experienced as a process which changes the perception of life. In participants' experience, spiritual support has a deeper and more positive impact than other psychosocial services. Research findings show the need for a more dynamic education process based on psychology and social sciences to enhance effective communication and interviewing skills.


Possible environmental origin of resistance of Aspergillus fumigatus to medical triazoles.
We reported the emergence of resistance to medical triazoles of Aspergillus fumigatus isolates from patients with invasive aspergillosis. A dominant resistance mechanism was found, and we hypothesized that azole resistance might develop through azole exposure in the environment rather than in azole-treated patients. We investigated if A. fumigatus isolates resistant to medical triazoles are present in our environment by sampling the hospital indoor environment and soil from the outdoor environment. Antifungal susceptibility, resistance mechanisms, and genetic relatedness were compared with those of azole-resistant clinical isolates collected in a previous study. Itraconazole-resistant A. fumigatus (five isolates) was cultured from the indoor hospital environment as well as from soil obtained from flower beds in proximity to the hospital (six isolates) but never from natural soil. Additional samples of commercial compost, leaves, and seeds obtained from a garden center and a plant nursery were also positive (four isolates). Cross-resistance was observed for voriconazole, posaconazole, and the azole fungicides metconazole and tebuconazole. Molecular analysis showed the presence of the dominant resistance mechanism, which was identical to that found in clinical isolates, in 13 of 15 environmental isolates, and it showed that environmental and clinical isolates were genetically clustered apart from nonresistant isolates. Patients with azole-resistant aspergillosis might have been colonized with azole-resistant isolates from the environment.


A rare case of Crohn's ileitis in a patient with constitutional mismatch repair deficiency.
Constitutional mismatch repair deficiency (CMMRD), a variant of Lynch syndrome, is a rare disease characterized by café-au-lait spots, oligopolyposis, glioblastoma and lymphoma. A 24-year-old male, under surveillance for CMMRD, developed Crohn's ileitis after total colectomy with end ileostomy for colorectal cancer and failed to respond to oral corticosteroids. The patient underwent induction and maintenance of remission with vedolizumab infusions. We report the first patient with CMMRD developing Crohn's disease. The choice of immunosuppressive therapy in these patients is challenging and needs to be made according to their risk for malignancy.


Insistence on sameness relates to increased covariance of gray matter structure in autism spectrum disorder.
Autism spectrum disorder (ASD) is characterized by atypical development of cortical and subcortical gray matter volume. Subcortical structural changes have been associated with restricted and repetitive behavior (RRB), a core component of ASD. Behavioral studies have identified insistence on sameness (IS) as a separable RRB dimension prominent in high-functioning ASD, though no simple brain-behavior relationship has emerged. Structural covariance, a measure of morphological coupling among brain regions using magnetic resonance imaging (MRI), has proven an informative measure of anatomical relationships in typical development and neurodevelopmental disorders. In this study, we use this measure to characterize the relationship between brain structure and IS. We quantified the structural covariance of cortical and subcortical gray matter volume in 55 individuals with high-functioning ASD using 3T MRI. We then related these structural metrics to individual IS scores, as assessed by the Repetitive Behavior Scale-Revised (RBS-R). We found that increased coupling among subcortical regions and between subcortical and cortical regions related to greater IS symptom severity. Most pronounced, the striatum and amygdala participated in a plurality of identified relationships, indicating a central role for these structures in IS symptomatology. These structural associations were specific to IS and did not relate to any of the other RRB subcomponents measured by the RBS-R. This study indicates that behavioral dimensions in ASD can relate to the coordination of development across multiple brain regions, which might be otherwise obscured using typical brain-behavior correlations. It also expands the structures traditionally related to RRB in ASD and provides neuroanatomical evidence supportive of IS as a separate RRB dimension. ClinicalTrials.gov NCT01031407.


Sesquiterpene emissions from pine trees--identifications, emission rates and flux estimates for the contiguous United States.
Biogenic volatile organic compound (BVOC) emissions were studied using vegetation enclosure experiments. Particular emphasis was given to sesquiterpene compounds (SQT), although monoterpenes (MT) were also characterized. SQT were detected in emissions from seven (out of eight) pine species that were examined. Thirteen SQT compounds were identified; the most abundant ones were beta-caryophyllene, alpha-bergamotene, beta-farnesene, and alpha-farnesene, with emission rates increasing exponentially with temperature. Regression analysis yielded exponential dependencies of both MT and SQT emissions on temperature of the form E = E0 x exp(beta(T - T0)). This resulted in SQT basal emission rates (E0 defined at T0 = 30 degrees C) ranging between <4 and 620 ng (carbon) gdw(-1) h(-1) (gdw = gram dry weight). The average value of the exponential temperature response factor beta for SQT emissions, taken from all experiments, was 0.17 degree C(-1), whereas the value for monoterpenes was 0.11 degrees C(-1). The average, total SQT emissions from pines were estimated to be 9, 16, and 29% of the MT emissions at 20, 30, and 40 degrees C respectively. The emission factors and beta-factors determined from these measurements were used to estimate pine tree MT and SQT emission distributions for the contiguous United States using MEGAN (model of emissions of gases and aerosols from nature, Guenther et al., 2006). SQT fluxes reaching 10-40 mg m(-2) for the month of July were estimated for extensive areas of most western and southern U.S. states.


Coping mechanisms during the COVID-19 pandemic and lockdown in metropolitan Johannesburg, South Africa: A qualitative study.
The COVID-19 pandemic has caused prolonged stress on numerous fronts. While the acute health impacts of psychosocial stress due to the pandemic are well-documented, less is known about the resources and mechanisms utilized to cope in response to stresses during the pandemic and lockdown. The aim of this study was to identify and describe the coping mechanisms adults utilized in response to the stressors of the COVID-19 pandemic during the 2020 South African lockdown. This study included adults (n = 47: 32 female; 14 male; 1 non-binary) from the greater Johannesburg region in South Africa. Interviews with both closed and open-ended questions were administered to query topics regarding the COVID-19 pandemic. Data were coded and thematically analyzed to identify coping mechanisms and experiences. Adults engaged in a variety of strategies to cope with the pandemic and the ensued lockdown. The ability to access or engage in multiple coping mechanisms were either enhanced or constrained by financial and familial situations. Participants engaged in seven major coping mechanisms: interactions with family and friends, prayer and religion, staying active, financial resources, mindset reframing, natural remedies, and following COVID-19 prevention protocols. Despite the multiple stressors faced during the pandemic and lockdown, participants relied on multiple coping strategies which helped preserve their well-being and overcome pandemic-related adversity. The strategies participants engaged in were impacted by access to financial resources and family support. Further research is needed to examine the potential impacts these strategies may have on people's health.


Molecular characterisation of ninein, a new coiled-coil protein of the centrosome.
We describe the cDNA cloning of ninein, a novel component of centrosomes. In the mouse, ninein is predicted to be an acidic protein (calculated pI of 4.8) with alternatively spliced forms of 245 kDa and 249 kDa that contain extensive regions of coiled-coil structure flanked by non-coiled ends. Other interesting features of this protein include an EF-hand-like domain, a potential GTP binding site and four leucine zipper domains. Specific polyclonal antisera were raised to two non-overlapping recombinant fragments of the protein and used to characterise the cellular distribution of ninein. Immunofluorescence and immunoelectron microscopy experiments with macrophage-like cells, Mm1, showed that ninein is localised specifically in the pericentriolar matrix of the centrosome. Studies with NIH3T3 fibroblasts demonstrated that ninein is associated with the centrosome throughout the cell cycle and can also be detected within nuclei at interphase. At mitosis ninein was also observed in association with the mitotic spindle. Immunocytochemical staining of mouse tissues showed that ninein was expressed in a heterogeneous fashion. Staining, if present, was always consistent with a centrosomal localisation and was never associated with nuclei. Ninein provides a new molecular tool for analysing the structure and function of the centrosome.


Multiple pulse nanosecond laser induced damage study in LiB3O5 crystals.
Multiple pulse nanosecond laser induced damage in the bulk of LiB3O5 (LBO) crystals was investigated at 1064 nm, 532 nm and 355 nm. Scanning electron microscopy of cleaved damage sites confirmed the presence of different zones that have already been reported in the case of KH2PO4 (KDP). Multi pulse measurements reveal a strong decrease of the damage threshold with increasing pulse number at 1064 nm (fatigue effect). A weaker fatigue effect was observed at 532 nm and no fatigue effect was found at 355 nm. This observation is best explained by an inherently statistical light matter interaction generating laser induced damage. Finally, a polarization dependent damage threshold anisotropy was evidenced at all three wavelengths, being strongest at 1064 nm. The results indicate the importance of Li+ vacancy stabilized color centers for the damage mechanism.


Serum profiling of healthy aging identifies phospho- and sphingolipid species as markers of human longevity.
As centenarians well represent the model of healthy aging, there are many important implications in revealing the underlying molecular mechanisms behind such successful aging. By combining NMR metabonomics and shot-gun lipidomics in serum we analyzed metabolome and lipidome composition of a group of centenarians with respect to elderly individuals. Specifically, NMR metabonomics profiling of serum revealed that centenarians are characterized by a metabolic phenotype distinct from that of elderly subjects, in particular regarding amino acids and lipid species. Shot- gun lipidomics approach displays unique changes in lipids biosynthesis in centenarians, with 41 differently abundant lipid species with respect to elderly subjects. These findings reveal phospho/sphingolipids as putative markers and biological modulators of healthy aging, in humans. Considering the particular actions of these metabolites, these data are suggestive of a better counteractive antioxidant capacity and a well-developed membrane lipid remodelling process in the healthy aging phenotype.


The crystal structures of substrate and nucleotide complexes of Enterococcus faecium aminoglycoside-2''-phosphotransferase-IIa [APH(2'')-IIa] provide insights into substrate selectivity in the APH(2'') subfamily.
Aminoglycoside-2''-phosphotransferase-IIa [APH(2'')-IIa] is one of a number of homologous bacterial enzymes responsible for the deactivation of the aminoglycoside family of antibiotics and is thus a major component in bacterial resistance to these compounds. APH(2'')-IIa produces resistance to several clinically important aminoglycosides (including kanamycin and gentamicin) in both gram-positive and gram-negative bacteria, most notably in Enterococcus species. We have determined the structures of two complexes of APH(2'')-IIa, the binary gentamicin complex and a ternary complex containing adenosine-5'-(beta,gamma-methylene)triphosphate (AMPPCP) and streptomycin. This is the first crystal structure of a member of the APH(2'') family of aminoglycoside phosphotransferases. The structure of the gentamicin-APH(2'')-IIa complex was solved by multiwavelength anomalous diffraction methods from a single selenomethionine-substituted crystal and was refined to a crystallographic R factor of 0.210 (R(free), 0.271) at a resolution of 2.5 A. The structure of the AMPPCP-streptomycin complex was solved by molecular replacement using the gentamicin-APH(2'')-IIa complex as the starting model. The enzyme has a two-domain structure with the substrate binding site located in a cleft in the C-terminal domain. Gentamicin binding is facilitated by a number of conserved acidic residues lining the binding cleft, with the A and B rings of the substrate forming the majority of the interactions. The inhibitor streptomycin, although binding in the same pocket as gentamicin, is orientated such that no potential phosphorylation sites are adjacent to the catalytic aspartate residue. The binding of gentamicin and streptomycin provides structural insights into the substrate selectivity of the APH(2'') subfamily of aminoglycoside phosphotransferases, specifically, the selectivity between the 4,6-disubstituted and the 4,5-disubstituted aminoglycosides.


Predicting relapse after transsphenoidal surgery for Cushing's disease.
Although selective transsphenoidal surgery is an effective treatment for pituitary-dependent Cushing's syndrome the definition of cure as distinct from improvement is unclear. Complete tumor removal should be associated with very low serum cortisol levels because of long-term suppression of normal corticotrophs but the optimum timing of this investigation after surgery has not been established. Eleven consecutive patients with surgical and histological confirmation of a corticotroph adenoma removed at transsphenoidal surgery for proven Cushing's disease were studied with 0900 h serum cortisol levels at 5-14 days and 6-12 weeks postoperatively. Patients were maintained on hydrocortisone 10 mg three times daily (final dose at 1800 h) pending recovery of the hypothalamic-pituitary axis which was assessed by periodic short tetracosactrin tests and continued remission of the condition was confirmed by low dose dexamethasone suppression testing. All patients achieved clinical resolution of their disease but four subsequently developed biochemical evidence of recurrence with incomplete suppression on low dose dexamethasone testing at 2-48 months after surgery. These patients had 0900 h serum cortisol levels of 124, 95, 186, and 265 nmol/L at 5-14 days and 334, 52, 130, and 240 nmol/L at 6-12 weeks postoperatively. The remaining seven patients, who are in remission after 8-83 (median 24) months of follow-up, demonstrated 0900 h serum cortisol levels of 30-75 (median 31) nmol/L at 5-14 days but lower levels at 6-12 weeks (< 20 nmol/L in three patients and 22, 30, 30, and 33 nmol/L in the remainder). In this series, serum cortisol measurements at 6-12 weeks after transsphenoidal surgery for Cushing's disease are lower than levels obtained within 2 weeks of surgery and appear to give better discrimination of continuing remission; levels less than 35 nmol/L suggest a favorable long-term outcome.


Prey-Capture Strategies of Fish-Hunting Cone Snails: Behavior, Neurobiology and Evolution.
The venomous fish-hunting cone snails (Conus) comprise eight distinct lineages evolved from ancestors that preyed on worms. In this article, we attempt to reconstruct events resulting in this shift in food resource by closely examining patterns of behavior, biochemical agents (toxins) that facilitate prey capture and the combinations of toxins present in extant species. The first sections introduce three different hunting behaviors associated with piscivory: 'taser-and-tether', 'net-engulfment' and 'strike-and-stalk'. The first two fish-hunting behaviors are clearly associated with distinct groups of venom components, called cabals, which act in concert to modify the behavior of prey in a specific manner. Derived fish-hunting behavior clearly also correlates with physical features of the radular tooth, the device that injects these biochemical components. Mapping behavior, biochemical components and radular tooth features onto phylogenetic trees shows that fish-hunting behavior emerged at least twice during evolution. The system presented here may be one of the best examples where diversity in structure, physiology and molecular features were initially driven by particular pathways selected through behavior.


Congenital disorders of glycosylation: an update on defects affecting the biosynthesis of dolichol-linked oligosaccharides.
Defects in the biosynthesis of the oligosaccharide precursor for N-glycosylation lead to decreased occupancy of glycosylation sites and thereby to diseases known as congenital disorders of glycosylation (CDG). In the last 20 years, approximately 1,000 CDG patients have been identified presenting with multiple organ dysfunctions. This review sets the state of the art by listing all mutations identified in the 15 genes (PMM2, MPI, DPAGT1, ALG1, ALG2, ALG3, ALG9, ALG12, ALG6, ALG8, DOLK, DPM1, DPM3, MPDU1, and RFT1) that yield a deficiency of dolichol-linked oligosaccharide biosynthesis. The present analysis shows that most mutations lead to substitutions of strongly conserved amino acid residues across eukaryotes. Furthermore, the comparison between the different forms of CDG affecting dolichol-linked oligosaccharide biosynthesis shows that the severity of the disease does not relate to the position of the mutated gene along this biosynthetic pathway.


Addressing Missing Data in Patient-Reported Outcome Measures (PROMS): Implications for the Use of PROMS for Comparing Provider Performance.
Patient-reported outcome measures (PROMs) are now routinely collected in the English National Health Service and used to compare and reward hospital performance within a high-powered pay-for-performance scheme. However, PROMs are prone to missing data. For example, hospitals often fail to administer the pre-operative questionnaire at hospital admission, or patients may refuse to participate or fail to return their post-operative questionnaire. A key concern with missing PROMs is that the individuals with complete information tend to be an unrepresentative sample of patients within each provider and inferences based on the complete cases will be misleading. This study proposes a strategy for addressing missing data in the English PROM survey using multiple imputation techniques and investigates its impact on assessing provider performance. We find that inferences about relative provider performance are sensitive to the assumptions made about the reasons for the missing data.


On the front line: structural insights into plant-pathogen interactions.
Over the past decade, considerable advances have been made in understanding the molecular mechanisms that underpin the arms race between plant pathogens and their hosts. Alongside genomic, bioinformatic, proteomic, biochemical and cell biological analyses of plant-pathogen interactions, three-dimensional structural studies of virulence proteins deployed by pathogens to promote infection, in some cases complexed with their plant cell targets, have uncovered key insights into the functions of these molecules. Structural information on plant immune receptors, which regulate the response to pathogen attack, is also starting to emerge. Structural studies of bacterial plant pathogen-host systems have been leading the way, but studies of filamentous plant pathogens are gathering pace. In this Review, we summarize the key developments in the structural biology of plant pathogen-host interactions.


Child physical abuse: factors influencing the associations between self-reported exposure and self-reported health problems: a cross-sectional study.
Child physical abuse (CPA) is an extensive public health problem because of its associations with poor health outcomes. The aim of this study was to examine which of the background factors of CPA committed by a parent or other caregiver relates to self-reported poor health among girls and boys (13; 15 and 17 years old): perpetrator, last year exposure; severity and frequency; socioeconomic load and foreign background. In a cross-sectional study in a Swedish county (n = 8024) a path analysis was performed to evaluate a model where all background variables were put as predictors of three health-status variables: mental; physical and general health problems. In a second step a log linear analysis was performed to examine how the distribution over the health-status categories was different for different combinations of background factors. Children exposed to CPA reported poor health to a much higher extent than those who were not exposed. In the path analysis it was found that frequency and severity of abuse (boys only) and having experienced CPA during the last year, was significantly associated with poor health as well as socioeconomic load in the families. Foreign background was significantly negatively associated with all three health indicators especially for girls. Neither mother nor father as perpetrator remained significant in the path analysis, while the results from the log linear analyses showed that mother-abuse did in fact relate to poor general health and mental as well as physical health problems among boys and girls. Father-abuse was associated with poor mental health if severe abuse was reported. Poor mental health was also associated with mild father-abuse if exposure during the last year was reported. Despite the limitations that cross-sectional studies imply, this study provides new knowledge about factors associated with poor health among physically abused children. It describes details of CPA that have significant associations to different aspects of poor health and thus what needs to be addressed by professionals within mental health providers and social services. Understanding how different factors may contribute to different health outcomes for exposed children is important in future research and needs further studies.


Animal welfare: an aspect of care, sustainability, and food quality required by the public.
People feel that they have obligations to the animals that they use and show some degree of care behavior toward them. In addition, animal welfare is an aspect of our decisions about whether animal-usage systems are sustainable. A system that results in poor welfare is unsustainable because it is unacceptable to many people. The quality of animal products is now judged in relation to the ethics of production, including impact on the animal's welfare on immediate features and on consequences for consumers. Because genetic selection and management for high productivity may lead to more disease and other aspects of poor welfare, consumers demand some major changes in animal-production systems. In teaching animal welfare, a clear definition that can be related to other concepts such as needs, health, and stress is needed. The methodology for the scientific assessment of animal welfare has developed rapidly in recent years and has become a major scientific discipline. No veterinary degree course should be approved unless a full course on the science of animal welfare and relevant aspects of ethics and law have been taught. Each country should have a national advisory committee on animal-welfare science, made up of independent scientists, including veterinarians, who can write impartial reviews of the state of scientific knowledge.


Orally co-administrated oleo-gum resin of Commiphora myrrha decreases the bioavailability of cyclosporine A in rats.
Cyclosporine A is a narrow therapeutic indexed immunosuppressant used after organ transplantation. Several herbs have been reported to alter its pharmacokinetics. Myrrh, dried oleogum resin obtained from Commiphora myrrha (Burseraceae) has been used for many common ailments. The present study was carried out to investigate the effect of myrrh on the pharmacokinetics of cyclosporine A. The rats of the control group received 60 mg/kg, p.o. cyclosporine A, and blood samples were collected at predetermined time intervals. Rats of the test group were treated with an aqueous suspension of myrrh (380 mg/kg p.o.) for eight days and on 8th day a single dose of cyclosporine A was administered to the treated group after 1 h of myrrh administration. Blood samples were drawn at predetermined time points and the drug was analyzed in whole blood by using H-Class UPLC-TQD. Pharmacokinetic profiles of control and test group were compared. Statistically significant differences were observed between the pharmacokinetic parameters of control and treated groups. In the myrrh treated group, the AUC(0-t) and C(max) of cyclosporine A was decreased by about 45% and 48%, respectively. The time to reach maximum concentration (T(max)) remained almost unchanged in both groups. Results indicated that the bioavailability of cyclosporine A was reduced by about 45% when co-administered with myrrh. This observation suggests that concurrent consumption of myrrh and cyclosporine A should be avoided. To confirm the clinical relevance of these findings, P-gp and CYP3A based molecular investigations can be performed along with a well-planned clinical study.


Solar photovoltaic interventions have reduced rural poverty in China.
Since 2013, China has implemented a large-scale initiative to systematically deploy solar photovoltaic (PV) projects to alleviate poverty in rural areas. To provide new understanding of China's targeted poverty alleviation strategy, we use a panel dataset of 211 pilot counties that received targeted PV investments from 2013 to 2016, and find that the PV poverty alleviation pilot policy increases per-capita disposable income in a county by approximately 7%-8%. The effect of PV investment is positive and significant in the year of policy implementation and the effect is more than twice as high in the subsequent two to three years. The PV poverty alleviation effect is stronger in poorer regions, particularly in Eastern China. Our results are robust to alternative specifications and variable definitions. We propose several policy recommendations to sustain progress in China's efforts to deploy PV for poverty alleviation.


Eye development: stable cell fate decisions in insect colour vision.
The retina provides an example of a fundamental property of developing cells: cell fate decisions are stable. A recent paper reports a double negative feedback loop that leads to bistable fate decisions in the colour-detecting photoreceptors of Drosophila.


Triticum aestivum L. endoxylanase inhibitor (TAXI) consists of two inhibitors, TAXI I and TAXI II, with different specificities.
The Triticum aestivum L. endoxylanase inhibitor (TAXI) discovered by Debyser and Delcour [(1997) Eur. Pat. filed April 1997, published as WO 98/49278] and Debyser, Derdelinckx and Delcour [(1997) J. Am. Soc. Brew. Chem. 55, 153-156] seems to be a mixture of two different endoxylanase inhibitors, called TAXI I and TAXI II. By using Aspergillus niger as well as Bacillus subtilis endoxylanases for assaying inhibition activity, both inhibitors could be purified to homogeneity from wheat (Triticum aestivum L., var. Soissons). TAXI I and TAXI II have similar molecular structures. They both have a molecular mass of approx. 40.0 kDa, are not glycosylated and occur in two molecular forms, i.e. a non-proteolytically processed one and a proteolytically processed one. However, the pI of TAXI II (at least 9.3) is higher than that of TAXI I (8.8). TAXI I and TAXI II clearly show different inhibition activities towards different endoxylanases. The N-terminal amino acid sequences of both inhibitors show a high degree of identity, which might indicate that there is an evolutionary relationship between them.


Incomplete reduction of branchial clefts in Mutton Merino lambs.
Congenital malformations of the branchial arches, clefts and grooves have not been previously reported in sheep. These defects may be due to infectious agents (especially viruses), toxins or genetic abnormalities. Defects were reported in two of a set of quadruplet lambs born prematurely to an eight-tooth Mutton Merino ewe. The lambs weighed between 2.0 kg and 2.5 kg; this is below the normal expected birth weight of 3.5 kg for quadruplet lambs, below which viability is compromised. The firstborn lamb was severely affected by bilateral oroauricular fistulae. The second lamb was unilaterally affected on the right, less severely than the first. The third lamb was normal and the fourth was mummified. The occurrence of another case in this small flock almost a decade earlier indicates that there could be genetic involvement.


Treatment of primary progressive multiple sclerosis.
Primary progressive multiple sclerosis (MS) is primarily a neurodegenerative phenotype of MS. It is characterized by a predominant progressive disease course that largely determines the disability progression rather than the rare early superimposed relapses. Therefore, frustratingly, the immunomodulatory and immunosuppressive approaches have failed to have any meaningful impact on the disability progression in primary-progressive MS. Some benefit can still be obtained in select patients where notable number of superimposed relapses or significant ongoing subclinical magnetic resonance imaging activity may allow for immunomodulatory agents to work. Symptomatic treatment for MS should also not be overlooked in this population for impact on quality of life. Future work will likely focus more on the remyelinating and regenerative strategies to help this group of patients. Unfortunately, the lack of any warning symptoms hampers any future prevention trial in this population. However, inferring from previous studies in secondary progressive MS, a global effort to eliminate initiation of tobacco smoking in teenagers or young adults may also delay the onset of a progressive disease course in MS.


Improving oral health through understanding the meaning of health and disease in a Chinese culture.
Advances in modern technology have undoubtedly been of benefit to dentistry in its preventive and curative activities for populations worldwide. Studies, mainly from Western societies, show nonetheless that utilisation of services or adoption of preventive methods are not taken up at satisfactory levels. The focus of this paper is on the special problems in a Chinese society where major differences exist between the traditional lay health concepts and the modernised professional ones. Traditional Chinese health concepts comprise the balance between and interaction of 'Yin' and 'Yang', the vital forces in both the universe and in man's body, as the basis for maintaining health. To restore a disturbed equilibrium of vital forces, which will manifest itself as disease, a range of dietary products are available in Hong Kong, for example cooling herb tea to reduce fire from the stomach producing gum disease. Seeking dental treatment from dentists could be delayed or made 'unnecessary' due to reliance on home remedies. This could reduce the effectiveness of any planned health education activities and dental services available for improving the oral health status of the community according to prevailing Western concepts. An understanding of the cultural meaning of dental disease is fundamental to changing beliefs and attitudes of people in order to improve the oral health of the community.


Streaming Physiological Data: General Public Perceptions of Secondary Use and Application to Research in Neonatal Intensive Care.
High speed physiological data represents one of the most untapped resources in healthcare today and is a form of Big Data. Physiological data is captured and displayed on a wide range of devices in healthcare environments. Frequently this data is transitory and lost once initially displayed. Researchers wish to store and analyze these datasets, however, there is little evidence of any engagement with citizens regarding their perceptions of physiological data capture for secondary use. This paper presents the findings of a self-administered household survey (n=165, response rate = 34%) that investigated Australian and Canadian citizens' perceptions of such physiological data capture and re-use. Results indicate general public support for the secondary use of physiological streaming data. Discussion considers the potential application of such data in neonatal intensive care contexts in relation to our Artemis research. Consideration of the perceptions of secondary use of the streaming data as early as possible will assist in building appropriate use models, with a focus on parents in the neonatal context.


Regional extent of an ecosystem engineer: earthworm invasion in northern hardwood forests.
The invasion of exotic earthworms into northern temperate and boreal forests previously devoid of earthworms is an important driver of ecosystem change. Earthworm invasion can cause significant changes in soil structure and communities, nutrient cycles, and the diversity and abundance of herbaceous plants. However, the regional extent and patterns of this invasion are poorly known. We conducted a regional survey in the Chippewa and Chequamegon National Forests, in Minnesota and Wisconsin, U.S.A., respectively, to measure the extent and patterns of earthworm invasion and their relationship to potential earthworm introduction sites. We sampled earthworms, soils, and vegetation in 20 mature, sugar maple-dominated forest stands in each national forest and analyzed the relationship between the presence of five earthworm taxonomic groups, habitat variables, and distance to the nearest potential introduction site. Earthworm invasion was extensive but incomplete in the two national forests. Four of the six earthworm taxonomic groups occurred in 55-95% of transects; however 20% of all transects were invaded by only one taxonomic group that has relatively minor ecological effects. Earthworm taxonomic groups exhibited a similar sequence of invasion found in other studies: Dendrobaena > Aporrectodea = Lumbricus juveniles > L. rubellus > L. terrestris. Distance to the nearest road was the best predictor of earthworm invasion in Wisconsin while distance to the nearest cabin was the best predictor in Minnesota. These data allow us to make preliminary assessments of landscape patterns of earthworm invasion. As an example, we estimate that 82% of upland mesic hardwood stands in the Wisconsin region are likely invaded by most taxonomic groups while only 3% are unlikely to be invaded at present. Distance to roads and cabins provides a coarse-scale predictor of earthworm invasion to focus stand-level assessments that will help forest managers better understand current and potential forest conditions and identify uninvaded areas that could serve as important refugia for plant species threatened by earthworm invasion.


Dura mater maintains rat cranial sutures in vitro by regulating suture cell proliferation and collagen production.
Craniosynostosis, the premature osseous obliteration of cranial vault sutures, can result from mutations in genes encoding components of growth factor signaling systems or the extracellular matrix (ECM). Little is known of the capacity of osteoprogenitor cells of the cranial sutures to divide or to synthesize ECM in situ. Osteoblasts derived from patients with prematurely fused sutures were reported to express alkaline phosphatase and osteocalcin at elevated levels, while proliferating at a rate comparable to control cells [DePollack et al., JBMR, 1996]; however, the suture osteoprogenitors, the population most likely to show proliferative abnormalities, were not present in the fused sutures used for this study. A model in which rat coronal sutures and associated bones develop normally in vitro, but in which sutures can be induced to fuse in the absence of dura mater, was used to examine cell proliferation and total protein synthesis in unfused sutures cultured in the presence of dura mater or in sutures induced to fuse in the absence of dura mater. Significantly increased cell proliferation was seen in suture cells prior to sutural obliteration, which returned to control levels as sutural fusion proceeded. Collagen synthesis in fusing sutures was elevated compared to non-fusing sutures and comparable to that seen in bone. Results indicated that in the absence of intercellular signals provided by the dura mater, suture cell proliferation increased initially, followed by increased synthesis of collagenous ECM within the suture and subsequent osseous obliteration of the suture. Thus factors originating in the dura mater affected suture cell proliferation and ECM production and were required for the maintenance of suture patency.


Effects of Nigella sativa on apoptosis and GABAA receptor density in cerebral cortical and hippocampal neurons in pentylenetetrazol induced kindling in rats.
We investigated the effects of Nigella sativa on apoptosis and gamma-aminobutyric acid (GABAA) receptor density in cerebral cortical and hippocampal neurons in a pentylenetetrazol (PTZ)-induced kindling model in rats. The PTZ kindling model was produced by injecting PTZ in subconvulsive doses to rats on days 1, 3, 5, 8, 10, 12, 15, 17, 19, 22 and 24 of the study into animals of PTZ treated (PTZ) and PTZ + N. sativa treated (PTZ + NS) groups. Clonic and tonic seizures were induced by injecting a convulsive dose of PTZ on day 26 of the study. Rats in the PTZ + NS group were treated also with a 10 mg/kg methanolic extract of N. sativa 2 h before each PTZ injection. Rats in the control group were treated with 4 ml/kg saline. The number of neurons that expressed GABAA receptors in the hippocampus and cerebral cortex of rats in the PTZ and PTZ + NS groups increased significantly. There was no significant difference in the number of GABAA receptors between the PTZ and PTZ + NS groups. GABAA receptor density of the neurons in the cerebral cortex, but not hippocampus, was increased in PTZ group compared to controls. We observed a significant increase in the number of apoptotic neurons in the cerebral cortex of rats of both the PTZ and PTZ + NS groups compared to controls. We observed a significant decrease in the number of the apoptotic neurons in the cerebral cortex of rats in the PTZ + NS group compared to the PTZ group. N. sativa treatment ameliorated the PTZ induced neurodegeneration in the cerebral cortex as reflected by neuronal apoptosis and neuronal GABAA receptor frequency.


Infant mortality rates according to socioeconomic status in a Brazilian city.
Data from municipal databases can be used to plan interventions aimed at reducing inequities in health care. The objective of the study was to determine the distribution of infant mortality according to an urban geoeconomic classification using routinely collected municipal data. All live births (total of 42,381) and infant deaths (total of 731) that occurred between 1994 and 1998 in Ribeirão Preto, Brazil, were considered. Four different geoeconomic areas were defined according to the family head's income in each administrative urban zone. The trends for infant mortality rate and its different components, neonatal mortality rate and post-neonatal mortality rate, decreased in Ribeirão Preto from 1994 to 1998 (chi-square for trend, p<0.05). These rates were inversely correlated with the distribution of lower salaries in the geoeconomic areas (less than 5 minimum wages per family head), in particular the post-neonatal mortality rate (chi-square for trend, p<0.05). Finally, the poor area showed a steady increase in excess infant mortality. The results indicate that infant mortality rates are associated with social inequality and can be monitored using municipal databases. The findings also suggest an increase in the impact of social inequality on infant health in Ribeirão Preto, especially in the poor area. The monitoring of health inequalities using municipal databases may be an increasingly more useful tool given the continuous decentralization of health management at the municipal level in Brazil.


A third novel locus for primary autosomal recessive microcephaly maps to chromosome 9q34.
Primary autosomal recessive microcephaly is a clinical diagnosis of exclusion in an individual with a head circumference >/=4 SDs below the expected age-and-sex mean. There is associated moderate mental retardation, and neuroimaging shows a small but structurally normal cerebral cortex. The inheritance pattern in the majority of cases is considered to be autosomal recessive. Although genetic heterogeneity for this clinical phenotype had been expected, this has only recently been demonstrated, with the mapping of two loci for autosomal recessive primary microcephaly: MCPH1 at 8p and MCPH2 at 19q. We have studied a large multiaffected consanguineous pedigree, using a whole-genome search, and have identified a third locus, MCPH3 at 9q34. The minimal critical region is approximately 12 cM, being defined by the markers cen-D9S1872-D9S159-tel, with a maximum two-point LOD score of 3.76 (recombination fraction 0) observed for the marker D9S290.


Breeding birds on small islands: island biogeography or optimal foraging?
1. We test MacArthur and Wilson's theory about the biogeography of communities on isolated habitat patches using bird breeding records from 16 small islands off the coasts of Britain and Ireland. 2. A traditional examination of patterns of species richness on these islands suggests that area and habitat diversity are important predictors, but that isolation and latitude have a negligible impact in this system. 3. Unlike traditional studies, we directly examine the fundamental processes of colonization and local extinction (cessation of breeding), rather than higher-order phenomena such as species richness. 4. We find that many of MacArthur and Wilson's predictions hold: colonization probability is lower on more isolated islands, and extinction probability is lower on larger islands and those with a greater diversity of habitats. 5. We also find an unexpected pattern: extinction probability is much lower on more isolated islands. This is the strongest relationship in these data, and isolation is the best single predictor of colonization and extinction. 6. Our results show that examination of species richness alone is misleading. Isolation has a strong effect on both of the dynamic processes that underlie richness, and in this system, the reductions in both colonization and extinction probability seen on more distant islands have opposing influences on species richness, and largely cancel each other out. 7. We suggest that an appropriate model for this system might be optimal foraging theory, which predicts that organisms will stay longer in a resource patch if the distance to a neighbouring patch is large. If nest sites and food are the resources in this system, then optimal foraging theory predicts the pattern we observe. 8. We advance the hypothesis that there is a class of spatial systems, defined by their scale and by the taxon under consideration, at which decision-making processes are a key driver of the spatiotemporal dynamics. The appropriate theory for such systems will be a hybrid of concepts from biogeography/metapopulation theory and behavioural ecology.


Targeting of XIAP combined with systemic mesenchymal stem cell-mediated delivery of sTRAIL ligand inhibits metastatic growth of pancreatic carcinoma cells.
Disseminating tumors are one of the gravest medical problems. Here, we combine the tumor-specific apoptosis-inducing activity of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) with the ability of mesenchymal stem cells (MSCs) to infiltrate both tumor and lymphatic tissues to target primary tumors as well as disseminated cancer cells in a human pancreatic cancer mouse model. Furthermore, we targeted X-linked inhibitor of apoptosis protein (XIAP) by RNA interference (RNAi) inside the cancer cells to make use of the apoptosis sensitization as well the antimetastatic effect that is afforded by XIAP silencing. We generated MSCs, termed MSC.sTRAIL, that express and secrete a trimeric form of soluble TRAIL (sTRAIL). MSC.sTRAIL triggered limited apoptosis in human pancreatic carcinoma cells that were resistant to soluble recombinant TRAIL, which is most likely due to the enhanced effect of the direct, cell-mediated delivery of trimeric TRAIL. MSC.sTRAIL-mediated cell death was markedly increased by concomitant knockdown of XIAP by RNAi in the cancer cells. These findings were confirmed in xenograft models, in which tumors from the parental pancreatic carcinoma cells showed only growth retardation on treatment with MSC.sTRAIL, whereas tumors with silenced XIAP that were treated with MSC.sTRAIL went into remission. Moreover, animals with XIAP-negative xenografts treated with MSC.sTRAIL were almost free of lung metastasis, whereas animals treated with control MSCs showed substantial metastatic growth in the lungs. In summary, this is the first demonstration that a combined approach using systemic MSC-mediated delivery of sTRAIL together with XIAP inhibition suppresses metastatic growth of pancreatic carcinoma.


Hypoxic modulation of Ca2+ signaling in human venous endothelial cells. Multiple roles for reactive oxygen species.
The effects of hypoxia (pO2 approximately 25 mm Hg) on Ca2+ signaling stimulated by extracellular ATP in human saphenous vein endothelial cells were investigated using fluorimetric recordings from Fura-2 loaded cells. In the absence of extracellular Ca2+, ATP-evoked rises of cytosolic Ca2+ concentration ([Ca2+]i) because of mobilization from the endoplasmic reticulum (ER). These responses were reduced by prior exposure to hypoxia but potentiated during hypoxia. Hypoxia itself liberated Ca2+ from the ER, but unlike the effects of ATP this effect was not inhibited by blockade of the inositol trisphosphate receptor. By contrast, ryanodine blocked the effects of hypoxia but not those of ATP. Antioxidants abolished the effects of hypoxia but potentiated the effects of ATP. Inhibition of NADPH oxidase also augmented ATP-evoked responses but was without effect on hypoxia-evoked rises of [Ca2+]i. However, either uncoupling mitochondrial electron transport or inhibiting complex I markedly suppressed the actions of hypoxia yet exerted only small inhibitory effects on ATP-evoked rises of [Ca2+]i. Both hypoxia and ATP were able to activate capacitative Ca2+ entry. Our results indicate that hypoxia regulates intracellular Ca2+ signaling via two distinct pathways. First, it modulates agonist-evoked liberation of Ca2+ from the ER primarily through regulation of reactive oxygen species generation from NADPH oxidase. Second, it liberates Ca2+ from the ER via ryanodine receptors, an effect requiring mitochondrial reactive oxygen species generation. These findings suggest that local O2 tension is a major determinant of Ca2+ signaling in the vascular endothelium, a finding that is likely to be of both physiological and pathophysiological importance.


Caregiver adherence to follow-up after an emergency department visit for common pediatric illnesses: Impact on future ED use.
Access to primary care is often a problem for children living in urban areas and the rate of emergency department (ED) use can be high. For acute childhood illnesses, primary care follow-up is often recommended to prevent subsequent ED visits. We conducted an observational study of 455 children with common childhood illnesses, between 6 weeks and 8 years of age, presenting to 1 of 3 EDs, and discharged to the community. ED physicians recommended that the child visit their primary care physician within 1 to 4 days of discharge (ie, "short-term" follow-up). Caregivers were surveyed during the ED index visit and after discharge to assess primary care follow-up adherence. We collected data on child and caregiver characteristics, type and severity of illness at the ED index visit, and ED return visits in the 2-month period after the ED index visit. A total of 45.3% of caregivers adhered to short-term primary care follow-up. Short-term follow-up adherence was associated with greater ED use for the same illness over the subsequent 2 months (odds ratio = 2.97; 95% confidence interval, 1.31-6.72). Subsequent ED use was greatest for children with short-term primary care follow-up and: (1) prior ED use, (2) single caregivers, (3) mild severity illnesses at the ED index visit, or (4) younger children. ED use after the initial visit did not vary by type of illness or site. There was no evidence that primary care follow-up soon after an ED visit was associated with a lower rate of subsequent ED use for common pediatric illnesses.


Factors Influencing the Frequency of Emergency Department Utilization by Individuals with Substance Use Disorders.
This study assessed the characteristics of individuals with substance use disorders (ISUDs) according to their frequency of emergency department (ED) utilization, and examined which variables were associated with an increase in ED visits using Andersen's model. Data linkage of administrative databanks from three sources [addiction rehabilitation centre registry, physician billing systems, and hospital discharge databank] for 4526 ISUDs was performed. Predisposing, enabling and need factors associated with number of ED visits were determined using a negative binomial regression model and generalised estimating equations. The rate of ED utilization for this population was 9.6 %. Increased number of ED visits was associated with the following variables: older age, social fragmentation, number of consultations with general practitioners, number of consultations with psychiatrists, number of consultations with other types of physicians, alcohol abuse, drug abuse, schizophrenia, anxiety disorders, personality disorders, co-occurring substance dependence and mental disorders, co-occurring substance dependence and chronic physical disorders, and co-occurring mental health disorders and chronic physical disorders. By contrast, a diagnosis of substance dependence, co-occurring drug and alcohol abuse, and a co-occurring diagnosis of substance dependence with mental health and chronic physical disorders decreased ED visits. Efforts to reduce avoidable use of EDs should focus on chronic-disease management and other related strategies aimed at reinforcing services to ISUDs in the community, especially for ISUDs with a co-occurring diagnosis of either mental health disorders or chronic physical disorders.


Increase in tendon protein synthesis in response to insulin-like growth factor-I is preserved in elderly men.
Insulin-like growth factor-I (IGF-I) is known to be an anabolic factor in tendon, and the systemic levels are reduced with aging. However, it is uncertain how tendon fibroblasts are involved in tendon aging and how aging cells respond to IGF-I. The purpose of this study was to investigate the in vivo IGF-I stimulation of tendon protein synthesis in elderly compared with young men. We injected IGF-I in the patellar tendons of young (n = 11, 20-30 yr of age) and old (n = 11, 66-75 yr of age) men, and the acute fractional synthesis rate (FSR) of tendon protein was measured with the stable isotope technique and compared with the contralateral side (injected with saline as control). We found that tendons injected with IGF-I had significantly higher protein FSR compared with controls (old group: 0.018 ± 0.015 vs. 0.008 ± 0.008, young group: 0.016 ± 0.009 vs. 0.009 ± 0.006%/h, mean ± SE, P < 0.01). This increase in protein synthesis was seen in both young and old men, with no differences between age groups. The old group had markedly lower serum IGF-I levels compared with young (165 ± 17 vs. 281 ± 27 ng/ml, P < 0.01). In conclusion, local IGF-I stimulated tendon protein synthesis in both young and old men, despite lower systemic IGF-I levels in the old group. This could indicate that the changed phenotype in aging tendon is not caused by decreased fibroblast function.


Time-Dependent Mechanical Properties in Polyetherimide 3D-Printed Parts Are Dictated by Isotropic Performance Being Accurately Predicted by the Generalized Time Hardening Model.
The Fused-Deposition Modelling (FDM) technique has transformed the manufacturing discipline by simplifying operational processes and costs associated with conventional technologies, with polymeric materials being indispensable for the development of this technology. A lack of quantification of viscoelastic/plastic behavior has been noted when addressing FDM parts with Polyetherimide (PEI), which is currently being investigated as a potential material to produce functional end-products for the aerospace and health industry. Primary and secondary creep along with stress relaxation tests have been conducted on FDM PEI specimens by applying stresses from 10 to 40 MPa for 100 to 1000 min. Specimens were 3D printed by varying the part build orientation, namely XY, YZ, and XZ. Creep results were fitted to the Generalized Time Hardening equation (GTH), and then this model was used to predict stress relaxation behavior. FDM PEI parts presented an isotropic creep and stress relaxation performance. The GTH model was proven to have a significant capacity to fit viscoelastic/plastic performances for each single build orientation (r > 0.907, p < 0.001), as well as a tight prediction of the stress relaxation behavior (r > 0.998, p < 0.001). Averaged-orientation coefficients for GTH were also closely correlated with experimental creep data (r > 0.958, p < 0.001) and relaxation results data (r > 0.999, p < 0.001). FDM PEI parts showed an isotropic time-dependent behavior, which contrasts with previous publications arguing the significant effect of part build orientation on the mechanical properties of FDM parts. These findings are strengthened by the high correlation obtained between the experimental data and the averaged-coefficient GTH model, which has been proven to be a reliable tool to predict time-dependent performance in FDM parts.


Persistence of disseminated tumor cells after neoadjuvant treatment for locally advanced breast cancer predicts poor survival.
Presence of disseminated tumor cells (DTCs) in bone marrow (BM) and circulating tumor cells (CTC) in peripheral blood (PB) predicts reduced survival in early breast cancer. The aim of this study was to determine the presence of and alterations in DTC- and CTC-status in locally advanced breast cancer patients undergoing neoadjuvant chemotherapy (NACT) and to evaluate their prognostic impact. Bone marrow and peripheral blood were collected before NACT (BM1: n = 231/PB1: n = 219), at surgery (BM2: n = 69/PB2: n = 71), and after 12 months from start of NACT (BM3: n = 162/PB3: n = 141). Patients were included from 1997 to 2003 and followed until 2009 (or ten years follow-up). DTC- and CTC-status were determined by morphological evaluation of immunocytochemically detected cytokeratin-positive cells. The prognostic significance of DTCs/CTCs was assessed by univariate and multivariate Cox-regression analyses. Before NACT, DTCs and CTCs were detected in 21.2% and 4.9% of the patients, respectively. At surgery, 15.9% and 1.4% had DTC- and CTC-presence, compared to 26.5% and 4.3% at 12 months from start of NACT. Of patients for whom DTC results both before NACT and at 12 months were available, concordant results were observed in 68%, and 14 out of 65 had positive DTC-status at both time points. Presence of ≥ 1 DTC 12 months from start of NACT, but not at other time points, predicted reduced disease-free survival (DFS; HR 2.3, p = 0.003), breast cancer-specific survival (BCSS; HR 3.0, p < 0.001) and overall survival (OS; HR 2.8, p < 0.001). Before NACT, presence of ≥ 3 DTCs was also associated with unfavorable outcome, and reduced BCSS was observed for CTC-positive patients (HR 2.2, p = 0.046). In multivariate analysis, DTC status (</≥ 1 DTC) at 12 months after start of NACT remained as a prognostic factor for both DFS (HR 2.2, p = 0.005), BCSS (HR 2.6, p = 0.002) and OS (HR 2.6, p = 0.002). The survival for patients with change in DTC-status was determined by the DTC-status at 12 months. Presence of DTCs after NACT indicated high risk for relapse and death, irrespective of the DTC-status before treatment. The results supports the potential use of DTC analysis as a monitoring tool during follow up, for selection of patients to secondary treatment intervention within clinical trials.


Understanding the phase formation kinetics of nano-crystalline kesterite deposited on mesoscopic scaffolds via in situ multi-wavelength Raman-monitored annealing.
Kesterite, a highly promising photo-absorbing crystalline form of Cu2ZnSnS4 (CZTS), has been prepared via various routes. However, the lack of in-depth understanding of the dynamic phase formation process of kesterite leads to difficulties in optimizing its annealing conditions, hence its light harvesting performance. In this paper, in situ Raman monitored-annealing is applied to study the phase formation kinetics of nano-crystalline kesterite from a precursor deposited on a TiO2 mesoscopic scaffold. By performing in situ Raman annealing under different experimental conditions and wavelengths, several facts have been discovered: kesterite crystallization starts at as low as 170 °C, but after short time annealing at 300 °C followed by cooling, the initially formed kesterite is found to decompose. Annealing at 400 °C or higher is proven to be sufficient for stabilizing the kesterite phase. Annealing at the higher temperature of 500 °C is necessary though to promote a complete reaction and thus eliminate the parasitic copper tin sulfide (CTS) impurity intermediates identified at lower annealing temperatures. More importantly, the real-time temperature dependence of Raman peak intensity enhancement, shift and broadening for CZTS is established experimentally at 500 °C for 1 h, providing a valuable reference in future CZTS research. This work demonstrates the significance of using in situ Raman spectroscopy in elucidating the kesterite phase formation kinetics, a critical step towards full crystal phase control - a prerequisite for developing fully functional CZTS-based optoelectronic devices.


Comparison of metabolic fates of nicotinamide, NAD+ and NADH administered orally and intraperitoneally; characterization of oral NADH.
Since NADH has been implicated in medication for some symptoms and as a possible supplement for health, we characterized the metabolic fate of NADH orally given to mice by comparing with those of nicotinamide (Nam), NAD+ and NADH intraperitoneally or orally administered. Mice were individually housed in metabolic cages, and divided into two sets of four groups. Within each set, one group was intraperitoneally or orally administered saline and the other three groups received intraperitoneal or oral administration of a pharmacological dose of Nam, NAD+ or NADH (5 micromol/mouse). Twenty-four hour urine samples for the day before and days 1 to 4 after administration were collected and analyzed for Nam and its metabolites. When mice were administered saline alone, urinary excretion of Nam and its metabolites, such as nicotinamide N-oxide (Nam N-oxide), N1-methylnicotinamide (MNA), N1-methyl-2-pyridone-5-carboxamide (2-Py), and N1-methyl-4-pyridone-3-carboxamide (4-Py), was unchanged from day 0 to day 4. Intraperitoneal injection of Nam, NAD+ and NADH produced significant increases in urinary excretion of Nam and its metabolites. Similar results were obtained when Nam and NAD+ were given orally. On the other hand, oral administration of NADH did not bring about an increase in urinary excretion of Nam and its metabolites, suggesting that NADH in digestive organs has been decomposed to a compound(s) that cannot yield Nam. In fact, incubation of NADH at acidic pH to mimic the stomach resulted in rapid conversion of NADH to an unknown compound. Better understanding of the fate of oral NADH is needed for its therapeutic and supplemental use.


Pentraxin-3 is a novel biomarker of lung carcinoma.
Our objective was to validate the performance of three new candidate lung cancer biomarkers, pentraxin-3 (PTX3), human kallikrein 11 (KLK11), and progranulin. We analyzed by commercial ELISA, and with a blinded protocol, 422 samples from 203 patients with lung carcinoma, 180 individuals with high risk for lung cancer (heavy smokers), and 43 individuals with cancers other than lung. All samples were obtained from the Early Detection Research Network (Reference set A). We found that progranulin and KLK11 were not informative lung cancer biomarkers, with areas under the receiver operating characteristic curve (AUC; ROC), close to 0.50. However, PTX3 was an informative lung cancer biomarker, with considerable ability to separate lung cancer patients from high-risk controls. At 90% and 80% specificity, the sensitivities versus the high-risk control group were 37% and 48%, respectively. The discriminatory ability of PTX3 was about the same with all major subtypes and histotypes of lung cancer. The AUC of the ROC curves increased according to the disease stage, from 0.64 (stage I) to 0.72 (stage IV). PTX3, but not KLK11 or progranulin, is a new serum biomarker for lung carcinoma. Its diagnostic sensitivity and specificity is similar to other clinically used lung cancer biomarkers. More studies are needed to establish if PTX3 has clinical utility for lung cancer diagnosis and management.


Phase angle of entrainment in morning- and evening-types under naturalistic conditions.
Differences in morningness-eveningness among humans are commonly ascribed to circadian parameters, such as circadian period and responsivity to environmental time cues, as well as homeostatic sleep drive. Light is the primary synchronizer of the human biological clock, and if circadian differences exist between morning and evening types, they should have different phase angles of entrainment to the light/dark cycle; that is, morning and evening types should have different patterns of light exposure relative to endogenous circadian phase (ECP). When phase angle of entrainment is strictly defined as the relationship between a marker of ECP and the timing of light exposure, such differences have been demonstrated in the laboratory under controlled light/dark cycles and have recently been shown under conditions of spring and summer light exposure outside the laboratory, taking into account the variable intensity of light. Here, we report similar results from a large (n=66), diverse cohort of morning and evening types across the age span studied at all different times of the year. Differences between morning and evening types in light exposure relative to ECP, indicative of a difference in the phase angle of entrainment to the external light/dark cycle, were found. Specifically, evening types, compared to morning types, had a higher ratio of phase advancing to phase delaying by light. We interpret this as indicating a longer circadian period (tau) in evening types.


Natural pathogens of laboratory mice, rats, and rabbits and their effects on research.
Laboratory mice, rats, and rabbits may harbor a variety of viral, bacterial, parasitic, and fungal agents. Frequently, these organisms cause no overt signs of disease. However, many of the natural pathogens of these laboratory animals may alter host physiology, rendering the host unsuitable for many experimental uses. While the number and prevalence of these pathogens have declined considerably, many still turn up in laboratory animals and represent unwanted variables in research. Investigators using mice, rats, and rabbits in biomedical experimentation should be aware of the profound effects that many of these agents can have on research.


The mammalian SWI/SNF complex and the control of cell growth.
The mammalian SWI/SNF complex is a chromatin remodelling complex that uses the energy of ATP hydrolysis to facilitate access of transcription factors to regulatory DNA sequences. This complex, that was initially described as a co-factor for nuclear receptors, has recently been associated with the control of cell growth. Two of the subunits known as BRG-1 and brm can associate with the Retinoblastoma tumour suppressor gene product and co-operate with this protein for repression of E2F activity. In addition, expression of brm is frequently down-regulated upon cellular transformation and re-introduction of this protein into fibroblasts transformed by activated ras induces partial reversion of the transformed phenotype. Finally, the hSNF5/INI1 gene, encoding another subunit of the SWI/SNF complex, is subject to bi-allelic mutations in rhabdoid tumours, a very aggressive form of paediatric cancers. These observations provide a novel link between malignant transformation and chromatin remodelling machineries.


Menière's disease patients improve specific posturographic parameters following diagnostic intratympanic injection.
Evaluation of specific computerized posturographic parameters in patients with Menière's disease (MD) following the intratympanic injection of gadolinium, a contrast agent, used in radiological diagnosing. We have observed 12 adult patients with unilateral Menière's Disease subjected to inner ear magnetic resonance imaging (MRI) examination after intratympanic gadolinium injection (ITG). The diagnoses have been performed according to the guidelines of the American Academy of otolaryngology. Before and after 24 h the ITG, all patients were subjected to the clinical evaluation and computerized posturography (CP), in 4 conditions depending on open/closed eyes and with/without foam cushion under feet. After ITG, in the affected ear the MRI confirmed the endolymphatic hydrops revealing a thin or even disappeared perilymphatic space. The statokinesigram showed improvement of stability only with closed eyes on a foam cushion. The CP performed 24 h after the contrast intratympanic injection showed a significant reduction of Path Length and Confidence Ellipse Area, due to an improvement of vestibular function on static balance. This improvement could be directly dependent to intratympanic pressure modification mediated by volume of contrast liquid, by "columella effect". This study demonstrates the absence of vestibular damage in patients undergoing intratympanic gadolinium infiltration and confirms the relationship between intratympanic pressure and vestibular stability modifications providing positive evidences for an applicative use of CP as a functional assessment to better address diagnosis and follow-up in MD patients treated with intratympanic injections.


Determination of stereospecificity of benzo[a]pyrene diolepoxide-DNA antisera with site-specifically modified oligonucleotides.
Antisera developed against benzo[a]pyrene diolepoxide (BPDE)-DNA adducts are sensitive tools for detection of DNA adducts in human samples. All antisera currently used for biomonitoring studies were produced against DNA or guanosine modified with racemic anti-BPDE. Using a non-competitive enzyme-linked immunosorbent assay (ELISA), Venkatachalam and Wani (Carcinogenesis, 15, 565-572, 1994) recently tested polyclonal and monoclonal (5D2) antisera for cross-reactivity against oligonucleotides containing (+)-and (-)-trans-anti-BPDE-N2-guanine or N6-adenine adducts and showed different stereospecificity for the two antisera. Because of the importance of antiserum specificity in human biomonitoring studies, we have tested several monoclonal (Mab 5D11 and 5D2) and polyclonal (Pab #29) antisera developed against racemic anti-BPDE-DNA adducts, and Mab 8E11 developed against anti-BPDE-guanosine adducts. Stereoisomeric anti-BPDE-modified oligonucleotide adducts in the sequence 5'-d(CC-AT-CG*CTACC)-3' where G* = anti-BPDE-N2-dG with (+)-trans, (-)-trans, (+)-cis and (-)-cis adduct stereochemistry at the C10 position of anti-BPDE were tested by competitive ELISA. Two structurally related 5-methylchrysene diolepoxide adducts with G* = (+)- and (-)-trans-anti-5-MeCDE-N2-dG in the same oligonucleotide were also tested. While Mab5D2 had the highest affinity for the (-)-trans-anti-BPDE-modified oligomer, Mab 5D11 and 8E11 and Pab #29 recognized the (+)-trans-anti-BPDE-modified oligomer better than the (-)-trans-anti-BPDE modified oligomer. Mab 5D11 and Pab #29 recognized racemic anti-BPDE-modified DNA adducts better than trans-anti-BPDE-modified oligonucleotides; however, Mab 8E11 showed similar sensitivity to racemic anti-BPDE-DNA adducts and (+)- and (-)-trans-anti-BPDE-modified oligomers. All antisera exhibited lower reactivities with both 5-MeCDE modified oligomers. Because of their sensitive detection of (+)-trans-anti-BPDE-dG adducts, the primary adduct produced in vivo, Mab 8E11 and 5D11 and Pab #29 are appropriate for measurement of most adducts formed in humans.


Heparin molecularly imprinted polymer thin flm on gold electrode by plasma-induced graft polymerization for label-free biosensor.
Heparin, a highly sulfated glycosaminoglycan, is an important biomaterial having biological and therapeutic functionalities such as anticoagulation, regeneration, and protein stabilization. This study addresses a label-free quartz crystal microbalance (QCM) biosensor for heparin detection based on a macromolecularly imprinted polymer (MIP) as an artificial recognition element. We demonstrate the novel strategy for MIP in the form of thin film on a gold (Au) electrode with the plasma-induced graft polymerization (PIP) technique. The procedure of PIP is as follows: (i) Hexamethyldisiloxane plasma-polymerized thin film (PPF) as a pre-coating scaffold of active species for PIP (post-polymerization) is deposited on an Au electrode. (ii) The PPF/Au electrode is soaked in an water solution containing heparin (template), (2-(methacryloxy)-ethyl)trimethylammonium chloride acrylamide (functional monomer), acrylamide, and N,N-methylenebisacrylamide (crosslinker). Double bonds of monomer and crosslinker attacked by residually active species in pre-coating PPF cause radical chain reaction. Consequently, a growing polymer network of 20 nm thickness of PIP-MIP thin film is formed and grafted on the PPF/Au surface. (iii) The PIP-MIP/PPF/Au is washed by sodium chloride solution so as to remove the template. Non-imprinted polymer (NIP) is carried out like the same procedure without a template. The AFM, XPS, and QCM measurements show that the PIP process facilitates macromolecularly surface imprinting of template heparin where the template is easily removed and is rapidly rebound to PIP-MIP without a diffusional barrier. The heparin-PIP-MIP specifically binds to heparin compared with heparin analog chondroitin sulfate C (selective factor: 4.0) and a detectable range of heparin in the presence of CS (0.1 wt%) was 0.001-0.1 wt%. The PIP-NIP does not show selectivity between them. The evaluated binding kinetics are association (ka = 350 ± 100 M-1 s-1), dissociation (kd = (5.0 ± 2.0) × 10-4 s-1), and binding (KD = 1.3 ± 0.6 μM) constants, demonstrating that the PIP-MIP as a synthetic antibody can be applied to analytical chemistry.


Establishing a network of specialist Porphyria centres - effects on diagnostic activities and services.
The porphyrias are a heterogeneous group of rare metabolic diseases. The full spectrum of porphyria diagnostics is usually performed by specialized porphyria laboratories or centres. The European Porphyria Initiative (EPI), a collaborative network of porphyria centres formed in 2001, evolved in 2007 into the European Porphyria Network (EPNET), where participating centres are required to adhere to agreed quality criteria. The aim of this study was to examine the state and distribution of porphyria diagnostic services in 2009 and to explore potential effects of increased international collaboration in the field of these rare diseases in the period 2006-2009. Data on laboratory, diagnostic and clinical activities and services reported to EPI/EPNET in yearly activity reports during 2006 through 2009 were compared between reporting centres, and possible time trends explored. Thirty-five porphyria centres from 22 countries, five of which were non-European associate EPNET members, filed one or more activity reports to EPI/EPNET during the study period. Large variations between centres were observed in the analytical repertoire offered, numbers of analyses performed and type and number of staff engaged. The proportion of centres fulfilling the minimum criteria set by EPNET to be classified as a specialist porphyria centre increased from 80% to 94% during the study period. Porphyria services are unevenly distributed, and some areas are probably still lacking in specialized porphyria services altogether. However, improvements in the quality of diagnostic services provided by porphyria centres participating in EPI/EPNET were observed during 2006 through 2009.


Ritual genital cutting of female minors.
The traditional custom of ritual cutting and alteration of the genitalia of female infants, children, and adolescents, referred to as female genital mutilation or female genital cutting (FGC), persists primarily in Africa and among certain communities in the Middle East and Asia. Immigrants in the United States from areas in which FGC is common may have daughters who have undergone a ritual genital procedure or may request that such a procedure be performed by a physician. The American Academy of Pediatrics believes that pediatricians and pediatric surgical specialists should be aware that this practice has life-threatening health risks for children and women. The American Academy of Pediatrics opposes all types of female genital cutting that pose risks of physical or psychological harm, counsels its members not to perform such procedures, recommends that its members actively seek to dissuade families from carrying out harmful forms of FGC, and urges its members to provide patients and their parents with compassionate education about the harms of FGC while remaining sensitive to the cultural and religious reasons that motivate parents to seek this procedure for their daughters.


Older women with localized breast cancer: costs and survival rates increased across two time periods.
Significant national attention has been paid to the rising costs of cancer care. However, few studies have evaluated the association between trends in costs and survival outcomes. We used the Surveillance, Epidemiology, and End Results (SEER) Program-Medicare linked database to compare changes in costs and survival rates over time, among women ages 67-94 who were diagnosed with stage II or III breast cancer in 1994-96 or 2004-06. We found that median cancer-related costs increased from $12,335 to $17,396 among women with stage II disease, and from $18,107 to $32,598 among women with stage III disease. Although the median cost of breast surgery declined between the two study periods, the median cost of chemo- and radiation therapy increased substantially, leading to an overall rise in cancer-related costs. Meanwhile, adjusted overall five-year survival improved, from 67.8 percent to 72.5 percent for women with stage II disease and from 38.5 percent to 51.9 percent for those with stage III disease. These findings suggest that increases in cancer care costs have been accompanied by improved outcomes. Future work should identify opportunities to optimize efficiency in cancer care.