Prevalence of apical periodontitis and endodontic treatment in a Kosovar adult population.
Despite numerous studies on the prevalence of apical periodontitis (AP) and endodontic treatment in diverse geographical populations, there are currently no data on the prevalence of these conditions in populations of adults native to Kosovo. Therefore, little is known about how widespread these conditions are, and whether there is any correlation between root canal treatment and AP. The purpose of our research was to address this anomaly by investigating AP and endodontic treatment in an adult Kosovar population based on radiographic examination. The sample used for this study consisted of randomly selected individuals referred to the University Dentistry Clinical Center of Kosovo in the years 2006-2007. Orthopantomographs of 193 patients were evaluated. The periapical status of all teeth (with the exception of third molars) was examined according to Ørstavik's Periapical Index. The quality of the root canal filling was rated as 'adequate' or 'inadequate' based on whether all canals were filled, the depth of fill relative to the radiographic apex and the quality of compaction (absence/presence of voids). Data were analyzed statistically using the Chi-square test and calculation of odds ratios. Out of 4131 examined teeth, the prevalence of apical periodontitis (AP) and endodontic treatment was 12.3% and 2.3%, respectively. Of 95 endodontically-treated teeth, 46.3% were associated with AP. The prevalence of AP increased with age. The prevalence in subjects aged over 60 years old (20.2%) was higher than in other age groups. A statistically significant difference was found for the frequency of endodontically-treated teeth associated with AP in the 40-49 year age group (P < 0.001). Of some concern was the discovery that only 30.5% of the endodontically-treated teeth examined met the criteria of an acceptable root canal filling. Inadequately root-filled teeth were associated with an increased AP risk. The prevalence of AP and the frequency of endodontically-treated teeth with AP in this Kosovar population are higher than those found in other countries. Inadequate root canal fillings were associated with an increased prevalence of AP.


Training U.S. medical students to care for the chronically ill.
The growing prevalence of chronic illness has important implications for the training of all physicians. The authors assessed the degree to which undergraduate medical curricula explicitly address chronic care competencies selected through literature and expert review. In 2001, trained student assistants interviewed directors of required medical school courses (internal medicine, pediatrics, family practice, and ambulatory care clerkships; longitudinal care; and other relevant courses) at 16 representative U.S. medical schools sampled by geography, curriculum reform activity, and primary care orientation of graduates. Course directors were asked whether chronic care competencies were addressed using specific curricular methods (written objectives, course materials, observational evaluations, written/oral examinations, other required course activities), and to rate the importance (1 = not important; 5 = essential) of each competency for their course and for the overall undergraduate curriculum. All 70 eligible course directors responded. Of 49 chronic care competencies, 29 (59%) received mean importance ratings for a course of >/=3, but only 14 (29%) were addressed using two or more specific curricular methods. Course directors gave highest importance ratings (mean > 3.9) to screening for abuse, awareness of patients' sociocultural perspectives, and protecting patients' confidentiality. They gave lowest importance ratings (mean </= 2.6) to knowing strategies to maximize patients' potential, ability to discuss alternative information sources, and ability to assess equipment needs. Their importance ratings correlated only moderately with the number of curricular methods used to address each competency (r(2) = 0.27-0.80, p <.05). Course directors agreed about the importance of many competencies in chronic care but reported considerable variation in how they addressed competencies in their courses. Medical schools can improve training in chronic care by paying greater attention to specific methods for addressing important chronic care competencies.


Differences in characteristics between suicide cases of farm managers compared to those of farm labourers in Queensland, Australia.
Farmers constitute an occupation group at a heightened suicide risk compared to the general population. To date, research has tried to explain this peculiarity by identifying suicide risk factors that are common to the whole of the farming population. There are, however, indications that risk factors may be different for different sub-populations of farmers, such as younger/older farmers or farm managers/farm labourers. This study compared the characteristics of suicides by farm managers and farm labourers, while controlling for the effect of age. A review of two datasets, the Queensland Suicide Register and the National Coroners Information System, was conducted in which a total of 78 cases of farm managers and 69 cases of farm labourers were identified as a suicide during 2000-2009, Queensland, Australia. The main outcome measures included various demographic characteristics, circumstances related to death, health and mental health variables, and history of stressful life events. The two groups differed in marital status, living arrangements, ethnicity, physical and mental illness, alcohol and drug abuse, contact with a health professional prior to death, and specific life events such as relationship breakdown and recent/pending unemployment. The majority of these differences were not statistically significant once age was accounted for. However, differences in psychiatric variables and experience of a recent/pending unemployment remained significant. This study contributes towards better understanding of suicide among farmers in different job positions, and highlights the need for tailored suicide prevention initiatives that consider a combination of age- and job-specific suicide risk and protective factors among farmers.


The relationship of respiratory symptoms and lung function loss in cotton textile workers.
We explored the relationship between a number of respiratory symptoms or symptom complexes (e.g., cough, recent wheeze, byssinosis) and lung function in white cotton textile workers (CTW) and control subjects 45 yr of age or older. The CTW were studied in 1973 and in 1979 in Columbia, South Carolina, and the control subjects were studied in 1972 and in 1978 in Lebanon, Connecticut. The CTW had a high prevalence of byssinosis (18%) and other symptoms. For each symptom or symptom complex, among persons with the symptom, CTW had lower lung function on the average than did control subjects. Also, among persons without the particular symptom, CTW had lower lung function than did control subjects. This pattern also occurred for persons without any respiratory symptoms (i.e., asymptomatic). Moreover, persons (CTW or control subjects separately) reporting the presence of a particular symptom had, in general, lower lung function than did those without the symptom. Among nonsmokers, all these same patterns usually held, but differences were often not statistically significant because of the small numbers of persons with symptoms. We conclude that on a group basis, respiratory function is more impaired in CTW than in control subjects with respiratory symptoms, that respiratory function is more impaired in persons (CTW or control subjects) with symptoms than in those without, and that even asymptomatic CTW have significantly greater lung function impairment than do control subjects.


Declining Mortality in Patients With Acute Respiratory Distress Syndrome: An Analysis of the Acute Respiratory Distress Syndrome Network Trials.
There has been multiple advances in the management of acute respiratory distress syndrome, but the temporal trends in acute respiratory distress syndrome-related mortality are not well known. This study aimed to investigate the trends in mortality in acute respiratory distress syndrome patients over time and to explore the roles of daily fluid balance and ventilation variables in those patients. Secondary analysis of randomized controlled trials conducted by the Acute Respiratory Distress Syndrome Network from 1996 to 2013. Multicenter study involving Acute Respiratory Distress Syndrome Network trials. Patients with acute respiratory distress syndrome. None. Individual patient data from 5,159 acute respiratory distress syndrome patients (excluding the Late Steroid Rescue Study trial) were enrolled in this study. The crude mortality rate decreased from 35.4% (95% CI, 29.9-40.8%) in 1996 to 28.3% (95% CI, 22.0-34.7%) in 2013. By adjusting for the baseline Acute Physiology and Chronic Health Evaluation III, age, ICU type, and admission resource, patients enrolled from 2005 to 2010 (odds ratio, 0.61; 95% CI, 0.50-0.74) and those enrolled after 2010 (odds ratio, 0.73; 95% CI, 0.58-0.92) were associated with lower risk of death as compared to those enrolled before 2000. The effect of year on mortality decline disappeared after adjustment for daily fluid balance, positive end-expiratory pressure, tidal volume, and plateau pressure. There were significant trends of declines in daily fluid balance, tidal volume, and plateau pressure and an increase in positive end-expiratory pressure over the 17 years. Our study shows an improvement in the acute respiratory distress syndrome-related mortality rate in the critically ill patients enrolled in the Acute Respiratory Distress Syndrome Network trials. The effect was probably mediated via decreased tidal volume, plateau pressure, and daily fluid balance and increased positive end-expiratory pressure.


Preconditioning doses of NMDA promote neuroprotection by enhancing neuronal excitability.
Neuroprotection can be induced by low doses of NMDA, which activate both synaptic and extrasynaptic NMDA receptors. This is in apparent contradiction with our recent findings that extrasynaptic NMDA receptor signaling exerts a dominant inhibitory effect on prosurvival signaling from synaptic NMDA receptors. Here we report that exposure to low preconditioning doses of NMDA results in preferential activation of synaptic NMDA receptors because of a dramatic increase in action potential firing. Both acute and long-lasting phases of neuroprotection in the face of apoptotic or excitotoxic insults are dependent on this firing enhancement. Key mediators of synaptic NMDA receptor-dependent neuroprotection, phosphatidylinositol 3 kinase-Akt (PI3 kinase-Akt) signaling to Forkhead box subgroup O (FOXO) export and glycogen synthase kinase 3beta (GSK3beta) inhibition and cAMP response element-binding protein-dependent (CREB-dependent) activation of brain-derived neurotrophic factor (BDNF), can be induced only by low doses of NMDA via this action potential-dependent route. In contrast, NMDA doses on the other side of the toxicity threshold do not favor synaptic NMDA receptor activation because they strongly suppress firing rates below baseline. The classic bell-shaped curve depicting neuronal fate in response to NMDA dose can be viewed as the net effect of two antagonizing (synaptic vs extrasynaptic) curves: via increased firing the synaptic signaling dominates at low doses, whereas firing becomes suppressed and extrasynaptic signaling dominates as the toxicity threshold is crossed.


Significance of multiple HPV infection in cervical cancer patients and its impact on treatment response.
Human papilloma virus (HPV) is the major cause of invasive cervical cancer (ICC). The study aim was to determine the prevalence of HPV genotypes and to correlate HPV types with response to radiotherapy. A total of 43 cervical biopsies collected from sequentially enrolled patients were analyzed by DNA amplification with MY09/MY11 primers and sequenced to determine the HPV genotype. Samples with multiple infections were resolved by multiplex PCR, combined with array primer extension (APEX). HPV DNA was detected in 40 of 43 (93%) samples. Nine different HPVs, including the most common types -16 (53%) and -18 (13%) were detected. Other types were HPV 31, 33, 45, 52, 58, 66 and 68. Single HPV types were found in 33 of 40 samples (82%) and multiple types in 7 of 40 samples (18%). The following significant predictors were identified: a) HPV 58 was most significant (p=0.02), followed by HPV 18 (p=0.04) associated with lack of treatment response; b) tumor size (p=0.042) and treatment response (p=0.025) elicited association with HPV infection type; c) treatment failure were found to be nearly 5-fold higher in case of multiple infections than of single infection (57% versus 12%) (odds ratio = 9.66; 95% CI 1.6-6.00). d) Multiple HPV infections correlated most prominently with lack of treatment compared with single type infection (p=0.005). Hence, patients with multiple infections, large tumor size, and HPV 58 and/or 18, are at risk of treatment failure and need to be followed for response and suitable inter-ventions done for a favorable outcome.


Long-term treatment with imatinib results in profound mast cell deficiency in Ph+ chronic myeloid leukemia.
Although mast cells (MC) play an important role in allergic reactions, their physiologic role remains unknown. In mice, several models of MC-deficiency have been developed. However, no comparable human model is available. We examined the in vitro- and in vivo effects of the KIT-targeting drug imatinib on growth and development of human MC. Imatinib was found to inhibit stem cell factor (SCF)-induced differentiation of MC in long-term suspension cultures (IC50: 0.01 µM). Correspondingly, long-term treatment of chronic myeloid leukemia (CML) patients with imatinib (400 mg/day) resulted in a marked decrease in MC. In patients with continuous complete molecular response during therapy, bone marrow MC decreased to less than 5% of pre-treatment values, and also serum tryptase concentrations decreased significantly (pre-treatment: 32.0 ± 11.1 ng/ml; post-therapy: 3.4 ± 1.8, p<0.01). Other myeloid lineages, known to develop independently of KIT, were not affected by imatinib-therapy. Imatinib also produced a substantial decrease in MC-development in mice. However, no clinical syndrome attributable to drug-induced MC-deficiency was recorded in our CML patients. Together, imatinib suppresses MC production in vitro and in vivo. However, drug-induced MC depletion is not accompanied by adverse clinical events, suggesting that MC are less relevant to homeostasis in healthy tissues than we assumed so far.


Cerebral Blood Flow and Heart Rate Variability in Chronic Fatigue Syndrome: A Randomized Cross-Over Study.
Pain, fatigue, and concentration difficulties are typical features of chronic fatigue syndrome (CFS). The exact underlying mechanisms of these symptoms are still unknown, but available evidence suggests an important role for impaired pain modulation. As evidence also suggests that pain modulation is related to cardiovascular mechanisms, it seems logical to investigate whether cerebral blood flow (CBF) and heart rate variability (HRV) are altered in these patients. We aimed to investigate the role of the cardiovascular system in pain modulation and symptoms of CFS; the response of CBF and HRV to physical stress and their relation to the change in temporal summation (TS) of pressure pain and self-reported symptoms was evaluated. A controlled, randomized cross-over trial. University Hospital Brussels. Twenty CFS patients and 20 sedentary healthy controls were included in this study. In both of the groups, the change in TS of pressure pain, CBF (using transcranial Doppler), and HRV (using finger plethysmography) was examined during physical and emotional stress (to control for potential bias), as well as their association mutually and with self-reported symptoms of pain, fatigue, and concentrations difficulties. There was no significant interaction or group (F-values ranging from .100 to 1.862, P-values ranging from .754 to .181) effect in CBF or HRV parameters. HRV and CBF did change during physical exercise, but the changes did not differ between patients and controls. While pain scores during TS at the trapezius site reduced in the control group after the physical exercise protocol (P = .037), they did not change in the CFS group (P = .108), suggesting impaired pain modulation. There were no significant correlations between CBF, HRV, TS, and self-reported symptoms (all P-values of correlation analyses > .01). Although effect sizes were medium to large, the study sample was relatively low. Also, the mild nature of the exercise bout is discussable. Nonetheless, this mild exercise was able to provoke endogenous pain modulation in the control group, which endorsed a proper execution of the cycling exercise. Moreover, mild exercises are more applicable to daily physical activities in CFS patients than vigorous exercises. These results seem to refute the previously suggested alterations of CBF/HRV in CFS patients. These cardiovascular parameters appear not to explain pain before, during, and following exercise. Chronic pain, physical exercise, emotional stress, pain modulation, cardiovascular systems, temporal summation, pain pressure thresholds, transcranial Doppler, plethysmography.


Prediction of nitrogen metabolism-related genes in Anabaena by kernel-based network analysis.
Prediction of molecular interaction networks from large-scale datasets in genomics and other omics experiments is an important task in terms of both developing bioinformatics methods and solving biological problems. We have applied a kernel-based network inference method for extracting functionally related genes to the response of nitrogen deprivation in cyanobacteria Anabaena sp. PCC 7120 integrating three heterogeneous datasets: microarray data, phylogenetic profiles, and gene orders on the chromosome. We obtained 1348 predicted genes that are somehow related to known genes in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. While this dataset contained previously known genes related to the nitrogen deprivation condition, it also contained additional genes. Thus, we attempted to select any relevant genes using the constraints of Pfam domains and NtcA-binding sites. We found candidates of nitrogen metabolism-related genes, which are depicted as extensions of existing KEGG pathways. The prediction of functional relationships between proteins rather than functions of individual proteins will thus assist the discovery from the large-scale datasets.


The neuroscience of positive emotions and affect: Implications for cultivating happiness and wellbeing.
This review paper provides an integrative account regarding neurophysiological correlates of positive emotions and affect that cumulatively contribute to the scaffolding for happiness and wellbeing in humans and other animals. This paper reviews the associations among neurotransmitters, hormones, brain networks, and cognitive functions in the context of positive emotions and affect. Consideration of lifespan developmental perspectives are incorporated, and we also examine the impact of healthy social relationships and environmental contexts on the modulation of positive emotions and affect. The neurophysiological processes that implement positive emotions are dynamic and modifiable, and meditative practices as well as flow states that change patterns of brain function and ultimately support wellbeing are also discussed. This review is part of "The Human Affectome Project" (http://neuroqualia.org/background.php), and in order to advance a primary aim of the Human Affectome Project, we also reviewed relevant linguistic dimensions and terminology that characterizes positive emotions and wellbeing. These linguistic dimensions are discussed within the context of the neuroscience literature with the overarching goal of generating novel recommendations for advancing neuroscience research on positive emotions and wellbeing.


Negative energy balance with exercise in identical twins: plasma glucose and insulin responses.
The effects of long-term (93 days) negative energy balance on plasma glucose and insulin were investigated by means of exercise with constant energy intake in seven pairs of young sedentary male identical twins. Results showed a significant decrease in fasting (-24%, P < 0.02) and postprandial insulin (-16%, P < 0.05). Fasting and postprandial plasma glucose and glucagon were not modified. Mean glucose disposal rate measured during a euglycemic-hyperinsulinemic clamp performed in five twin pairs was increased by 34% (P = 0.13). No significant intrapair similarity was found for the responses of fasting and postprandial insulin levels and of glucose disposal rate. Changes in glucose disposal rate were significantly greater in high compared with low losers for computerized tomography-measured abdominal visceral fat (3.0 +/- 1.1 vs. 0.3 +/- 0.5 mg x kg(-1) x min(-1), P < 0.05) despite similar losses in total body fat. We conclude that 1) a long-term negative energy balance generated by exercise training significantly reduces plasma insulin levels, whereas insulin sensitivity tends to be improved; 2) training-induced changes in insulin sensitivity are associated with changes in abdominal visceral fat; and 3) data from this experiment conducted with a small number of twin pairs suggest that the genotype does not seem to be a major determinant of the changes in insulin levels and sensitivity brought about by negative energy balance with exercise.


B2O3/SiO2 substitution effect on structure and properties of Na2O-CaO-SrO-P2O5-SiO2 bioactive glasses from molecular dynamics simulations.
The effect of B2O3/SiO2 substitution in SrO-containing 55S4.3 bioactive glasses on glass structure and properties, such as ionic diffusion and glass transition temperature, was investigated by combining experiments and molecular dynamics simulations with newly developed potentials. Both short-range (such as bond length and bond angle) and medium-range (such as polyhedral connection and ring size distribution) structures were determined as a function of glass composition. The simulation results were used to explain the experimental results for glass properties such as glass transition temperature and bioactivity. The fraction of bridging oxygen increased linearly with increasing B2O3 content, resulting in an increase in overall glass network connectivity. Ion diffusion behavior was found to be sensitive to changes in glass composition and the trend of the change with the level of substitution is also temperature dependent. The differential scanning calorimetry (DSC) results show a decrease in glass transition temperature (Tg) with increasing B2O3 content. This is explained by the increase in ion diffusion coefficient and decrease in ion diffusion energy barrier in glass melts, as suggested by high-temperature range (above Tg) ion diffusion calculations as B2O3/SiO2 substitution increases. In the low-temperature range (below Tg), the Ea for modifier ions increased with B2O3/SiO2 substitution, which can be explained by the increase in glass network connectivity. Vibrational density of states (VDOS) were calculated and show spectral feature changes as a result of the substitution. The change in bioactivity with B2O3/SiO2 substitution is discussed with the change in pH value and release of boric acid into the solution.


Poly[(dimethyl-formamide)(μ(4)-2,2'-sulfanediyldibenzoato)nickel(II)].
The title centrosymmetric dinuclear Ni(II) complex, [Ni(C(14)H(8)O(4)S)(C(3)H(7)NO)](n), was prepared via reaction of Ni(NO(3))(2)·6H(2)O and thio-salicylic acid, with H(2)O and dimethyl-formamide (DMF) as the mixed solvent. The central Ni(II) ion is five-coordinated by five O atoms from DMF and from the carboxyl-ate groups of the organic ligand. The symmetry-related coordination polyhedra inter-link into centrosymmetric dimeric units and these, in turn, are linked into infinite chains propagating parallel to [100].


Youth caught in violation of tobacco purchase, use, and possession laws: education versus fines.
Each day, thousands of children are caught for violation of tobacco purchase, use, and possession (PUP) laws. Little is known about their impact on violators; we do not know how the youth who are caught perceive these consequences or the effects they have on their tobacco use. Moreover, many communities are beginning to use brief tobacco education programs as a diversion from the normal processing of PUP law violators (i.e., fining the youth violator) without knowing the consequences of these classes. Consequently, it is important to review the literature and studies that have evaluated the effects of civic fines versus tobacco education as a consequence for PUP law violations. A consolidation of this information along with a presentation of pilot data on this issue might suggest areas of needed future research as well as help policy officials make decisions about best practices in their communities regarding these types of laws.


Easy computation of the Bayes factor to fully quantify Occam's razor in least-squares fitting and to guide actions.
The Bayes factor is the gold-standard figure of merit for comparing fits of models to data, for hypothesis selection and parameter estimation. However, it is little-used because it has been considered to be subjective, and to be computationally very intensive. A simple computational method has been known for at least 30 years, but has been dismissed as an approximation. We show here that all three criticisms are misplaced. The method should be used to complement and augment all least-squares fitting, because it can give very different, and better outcomes than classical methods. It can discriminate between models with equal numbers of parameters and equally good fits to data. It quantifies the Occam's Razor injunction against over-fitting, and it demands that physically-meaningful parameters rejected by classical significance testing be included in the fitting, to avoid spurious precision and incorrect values for the other parameters. It strongly discourages the use of physically-meaningless parameters, thereby satisfying the Occam's Razor injunction to use existing entities for explanation rather than multiplying new ones. More generally, as a relative probability, the Bayes factor combines naturally with other quantitative information to guide action in the absence of certain knowledge.


The C allele of the reactive oxygen species modulator 1 (ROMO1) polymorphism rs6060566 is a biomarker predicting coronary artery stenosis in Slovenian subjects with type 2 diabetes mellitus.
We aimed to examine the role of the rs6060566 polymorphism of the reactive oxygen species modulator 1 (ROMO1) gene in the development of myocardial infarction (MI) in Caucasians with type 2 diabetes (T2DM). A total of 1072 subjects with T2DM were enrolled in this cross-sectional case-control study: 335 subjects with MI and 737 subjects without clinical signs of coronary artery disease (CAD). The genetic analysis of the rs6060566 polymorphism was performed in all subjects. To assess the degree of coronary artery obstruction, a subpopulation of 128 subjects with T2DM underwent coronary computed tomography angiography. Next, endarterectomy samples were obtained during myocardial revascularization from diffusely diseased coronary arteries in 40 cases, which were analysed for ROMO1 expression according to their genotype. There were no statistically significant associations between different genotypes or alleles of the rs6060566 polymorphism and MI in subjects with T2DM. The carriers of the C allele of the ROMO1 rs6060566 had a threefold increased likelihood of having 50-75% coronary artery stenosis (Adjusted OR = 3.27, 95% CI 1.16-9.20). Subjects with two affected coronary arteries had a 3.72 fold higher prevalence of MI (OR = 3.72, 95% CI 1.27-10.84). With CAD in LMCA or LAD, MI prevalence was about 3.5-fold higher (p = 0.07 for LMCA and p = 0.01 for LAD). Furthermore, the carriers of the rs6060566 C allele showed higher number of positive cells for ROMO1 expression in endarterectomy samples of coronary arteries. According to our study, the rs6060566 polymorphism of the ROMO1 gene is not a risk factor for MI in Caucasians with T2DM. However, we found that subjects carrying the C allele were at a 3.27-fold increased risk of developing severe CAD compared with those who had non-obstructive CAD. Moreover, C allele carriers showed a statistically higher number of cells positive for ROMO1 compared with T allele carriers in coronary endarterectomy samples.


Dry active turbulence in a model for microtubule-motor mixtures.
We study the dynamics and phase behaviour of a dry suspension of microtubules and molecular motors. We obtain a set of continuum equations by rigorously coarse graining a microscopic model where motor-induced interactions lead to parallel or antiparallel ordering. Through numerical simulations, we show that this model generically creates either stable stripes, or a never-settling pattern where stripes periodically form, rotate and then split up. We derive a minimal model which displays the same instability as the full model, and clarifies the underlying physical mechanism. The necessary ingredients are an extensile flux arising from microtubule sliding and an interfacial torque favouring ordering along density gradients. We argue that our minimal model unifies various previous observations of chaotic behaviour in dry active matter into a general universality class.


Inhibition of amplification of Chlamydia trachomatis plasmid DNA by the ligase chain reaction associated with female urines.
OBJECTIVE: To determine the rate of inhibition of ligase chain reaction (LCR) amplification of Chlamydia trachomatis plasmid DNA in urines from pregnant and non-pregnant women, to correlate inhibitors with urinary substances through urinalysis, and to explore methods of removing inhibitors from urine. METHODS: First-void urines (FVUs) from 287 non-pregnant and 101 pregnant women (15--40 years of age) in Hamilton, Canada were tested for urinary factors by urinalysis and for C. trachomatis by LCR. A sample of each urine was 'spiked' with a positive control containing approximately one infectious unit of C. trachomatis L2 serovar. RESULTS: Complete inhibition of DNA amplification was observed in nine of 287 (3.1%) urines from non-pregnant women and in one of 101 (1%) who were pregnant. Partial inhibition (> 20% reduction in signal) was observed in an additional five of 388 (1.3%) urines. Nine urines contained C. trachomatis plasmid DNA, but none of these contained inhibitors of LCR. By multivariate analysis, the only substances in urine which were significantly and independently related to LCR inhibition were nitrites. Holding the urines at 4C or minus sign70C overnight reduced the numbers which were inhibitory by approximately 50%. Diluting the urines 1:10 or extracting with phenol/chloroform removed all inhibition and retained amplifiable DNA. CONCLUSIONS: We conclude that urinary inhibitors of C. trachomatis DNA amplification by LCR are minimal and appear to be related to the presence of nitrites. We anticipate that most would disappear in transit or during processing and that a positive control for every urine is not warranted, considering the added cost and work associated with duplicate testing.


Evaluating Universal Design in Built Environments - A Scoping Project.
This project aimed to scope existing methods of evaluating the application of universal design to built environments and to explore relevant knowledge of key stakeholders, such as architects, access consultants and people who experience disability. The project commenced in 2017. Ethics approval was gained and a mixed methods approach was employed. Methods of data collection included electronic survey and in-depth interview. Early survey findings are reported in this paper. A descriptive approach was used to analyse quantitative data. A total of 157 survey responses were received from across Australia (83%) and internationally (16.6%). Preliminary findings indicate that most survey respondents (72%) had been involved in the process of applying universal design to the design of built environments. Although evaluating the application of universal design was rated as "extremely important" by 85% of respondents, only 36% had such experience. Of these, 74% had used specific tools for this purpose. Non-standardised checklists and access audits were the most frequently used and preferred tools. Overall, stakeholders perceived themselves to have 'some knowledge' on universal design theory and application. This project offers insight into how universal design is understood and applied to the design of built environments. Findings suggest that evaluation is less common than application and that there is a need to strengthen existing methods of evaluation to provide greater detail on universal design processes and outcomes.


Application of raisin extracts as preservatives in liquid bread and bread systems.
Interest in natural ingredients with multifunctions in food has led to the investigation of the antimicrobial activity of raisins, a traditional baking ingredient. Water and 60% ethanol extracts, and raisin juice concentrate (RJC, a commercial food ingredient) were assessed for their ability to inhibit the growth of ropy-bread-causing Bacillus species. The ability of these additives and raisin paste were tested for antifungal activity in liquid bread model system and bread. In the liquid bread model, concentrations as low as 20 and 36 mg crude extract or RJC per gram media significantly (P < 0.05) reduced the populations of B. licheniformis and B. subtilis, respectively. A significant reduction in the population of Aspergillus flavus and Penicillium chrysogenum was achieved at 80 and 470 mg crude extract per gram media, respectively. Bread formulas containing 7.5% of a water extract (equivalent to 70 mg extract per gram dough) or RJC (70 mg/g dough) produced bread that had equivalent mold-free shelf life. The mean mold-free shelf life of the bread containing 7.5% water extract was 18.1 +/- 3.3 d at room temperature while the negative control was mold free for 9.4 +/- 2.4 d. The antifungal efficacy of the extracts in bread was equivalent to 0.24% calcium propionate in 21 d of storage. Doubling the concentration of the extract did not improve the mold-retarding property in bread. The bread containing raisin paste, the percentage of which in dough was equivalent to 15% raisin extract, exhibited a stronger antifungal activity than did the extracts in bread.


Cardiff acne disability index: cross-cultural translation in Korean and its relationship with clinical acne severity, pathological patterns, and general quality of life.
To culturally translate the cardiff acne disability index (CADI) into Korean, and to examine its relationship with clinical acne severity, pathological patterns, and general quality of life (QoL). The CADI was culturally and lin- guistically translated into Korean via translation, back-translation, and face validity test process. Two hundred and fifty-four Korean adolescents were asked to complete the Korean version of the CADI (K-CADI), the Phlegm Pattern, the Cold-Heat Pattern, and the Korean version of the General Health Questionnaires. A clinician estimated acne severity for the adolescents, using the Korean Acne Grading System. Finally, reliability and validity of the K-CADI was examined, and the relationships between acne severity, Phlegm, Cold, and Heat patterns, and QoL level were examined via pathway analysis. The K-CADI had satisfactory internal con- sistency (α = 0.827). The examination of construct validity indicated that the K-CADI had one factor (explaining 59.6% of the total variance). Pathway analysis showed satisfactory model fit (normal fit index = 0.960 and comparative fit index = 0.983), and acne-related QoL was determinant to Phlegm, Heat, and Cold patterns (0.13-0.27 of β), and Phlegm and Heat patterns lowered one's QoL level (0.17-0.34 of β). The K-CADI is a valid and reliable instrument. Phlegm and Heat patterns should be managed when treating acne since they have a moderating effect on general QoL aggravation.


Biological cluster mass spectrometry.
This article reviews the new physics and new applications of secondary ion mass spectrometry using cluster ion probes. These probes, particularly C(60), exhibit enhanced molecular desorption with improved sensitivity owing to the unique nature of the energy-deposition process. In addition, these projectiles are capable of eroding molecular solids while retaining the molecular specificity of mass spectrometry. When the beams are microfocused to a spot on the sample, bioimaging experiments in two and three dimensions are feasible. We describe emerging theoretical models that allow the energy-deposition process to be understood on an atomic and molecular basis. Moreover, experiments on model systems are described that allow protocols for imaging on biological materials to be implemented. Finally, we present recent applications of imaging to biological tissue and single cells to illustrate the future directions of this methodology.


Genome-Wide Association Studies Reveal the Genetic Basis of Ionomic Variation in Rice.
Rice (Oryza sativa) is an important dietary source of both essential micronutrients and toxic trace elements for humans. The genetic basis underlying the variations in the mineral composition, the ionome, in rice remains largely unknown. Here, we describe a comprehensive study of the genetic architecture of the variation in the rice ionome performed using genome-wide association studies (GWAS) of the concentrations of 17 mineral elements in rice grain from a diverse panel of 529 accessions, each genotyped at ∼6.4 million single nucleotide polymorphism loci. We identified 72 loci associated with natural ionomic variations, 32 that are common across locations and 40 that are common within a single location. We identified candidate genes for 42 loci and provide evidence for the causal nature of three genes, the sodium transporter gene Os-HKT1;5 for sodium, Os-MOLYBDATE TRANSPORTER1;1 for molybdenum, and Grain number, plant height, and heading date7 for nitrogen. Comparison of GWAS data from rice versus Arabidopsis (Arabidopsis thaliana) also identified well-known as well as new candidates with potential for further characterization. Our study provides crucial insights into the genetic basis of ionomic variations in rice and serves as an important foundation for further studies on the genetic and molecular mechanisms controlling the rice ionome.


Impact on visual function from light scattering and glistenings in intraocular lenses, a long-term study.
To investigate the impact on visual function from light scattering and glistenings in intraocular lenses (IOLs) in patients who had cataract surgery 10 years previously. Eye clinic, Norrlands university hospital, Umeå, Sweden. One hundred and three patients, who had phacoemulsification with implantation of Acrysof(®) MA60BM IOLs 10 years previously, were evaluated with best corrected visual acuity (VA), and low contrast visual acuity (LCVA) 10% and 2.5%. The light scattering from the IOLs was measured by Scheimpflug photography. The degree of glistenings was also quantified at the slit-lamp. Eyes with coexisting pathology that could affect VA and LCVA were excluded. The patients were divided into various groups according to the degree of light scattering and grade of glistenings. In two subsets of patients, paired data from the patients' eyes were analysed. It was not possible to detect any significant impact on visual function, best corrected visual acuity (BCVA) and LCVA 10% and 2.5% in eyes with a more pronounced light scattering or a higher grade of glistenings seen at the slit-lamp. The correlation between IOL dioptric power and both the total light scattering of the IOL, and the subjective grading of the intensity of the glistenings at the slit-lamp was statistically significant (r(P)=0.25; p=0.012; r(S)=0.23; p=0.019, respectively). Most patients in this case series operated 10 years previously had severe glistenings and a high level of light scattering from their intraocular lenses. No detectable impact on BCVA, LCVA 10% and 2.5% was found.


Clinical and radiological evaluation of ejaculatory duct obstruction.
Ejaculatory duct obstruction (EDO) is a rare but surgically correctable cause of male infertility. With the advent and increased use of transrectal ultrasonography and magnetic resonance imaging, abnormalities of the ejaculatory duct (ED) related to infertility have been diagnosed more frequently. Recently, with the increased awareness of functional obstruction of ED, reports have been focusing on the diagnosis of partial or functional EDO. We present 2 review of the ED pathologies, imaging modalities and treatment options.


Staining of E-selectin ligands on paraffin-embedded sections of tumor tissue.
The E-selectin ligands expressed by cancer cells mediate adhesion of circulating cancer cells to endothelial cells, as well as within tissue microenvironments important for tumor progression and metastasis. The identification of E-selectin ligands within cancer tissue could yield new biomarkers for patient stratification and aid in identifying novel therapeutic targets. The determinants of selectin ligands consist of sialylated tetrasaccharides, the sialyl Lewis X and A (sLeX and sLeA), displayed on protein or lipid scaffolds. Standardized procedures for immunohistochemistry make use of the antibodies against sLeX and/or sLeA. However, antibody binding does not define E-selectin binding activity. In this study, we developed an immunohistochemical staining technique, using E-selectin-human Ig Fc chimera (E-Ig) to characterize the expression and localization of E-selectin binding sites on paraffin-embedded sections of different cancer tissue. E-Ig successfully stained cancer cells with high specificity. The E-Ig staining show high reactivity scores in colon and lung adenocarcinoma and moderate reactivity in triple negative breast cancer. Compared with reactivity of antibody against sLeX/A, the E-Ig staining presented higher specificity to cancer tissue with better defined borders and less background. The E-Ig staining technique allows the qualitative and semi-quantitative analysis of E-selectin binding activity on cancer cells. The development of accurate techniques for detection of selectin ligands may contribute to better diagnostic and better understanding of the molecular basis of tumor progression and metastasis.


NMN alleviates radiation-induced intestinal fibrosis by modulating gut microbiota.
Radiation-induced intestinal fibrosis, a common complication of long-term survivors after receiving abdominal and pelvic radiotherapy, has no effective clinical drugs at present. Nicotinamide mononucleotide (NMN) has been reported to alleviate a variety of age-related diseases and has potential of regulating gut microbiota. The current study focuses on the role of gut microbiota in chronic radiation induced intestinal fibrosis, and investigates whether NMN plays a protective role in radiation-induced intestinal fibrosis as well as the impact of NMN on radiation-induced dysbiosis of gut microbiota. C57BL/6J mice received 15 Gy abdominal irradiation and NMN (300 mg/kg/day) supplement in drinking water. Feces were collected at 4- and 8-months post-irradiation and performed 16S rRNA sequencing to detect the gut microbiota. Colon tissues were isolated at 12 months after irradiation with or without NMN supplementation for histological analysis. We found that irradiation caused intestinal fibrosis, and altered the β diversity and composition of gut microbiota, while the gut microbiota was observed to be affected by time post-irradiation and age of mice. Long-term NMN supplementation alleviated intestinal fibrosis, and reshaped the composition and function of gut microbiota dysregulated by ionizing radiation (IR). In addition, Akkermansia muciniphila, a promising probiotic, and metabolism-related pathways, such as Biosynthesis of other secondary metabolites and Amino acid metabolism, were more abundant after NMN treatment in irradiated mice. IR has a long-term effect on the gut microbiota and NMN supplementation can alleviate radiation induced intestinal fibrosis by reshaping the composition of gut microbiota and regulating the metabolic function of the microorganism.


The N-terminal ricin propeptide influences the fate of ricin A-chain in tobacco protoplasts.
The plant toxin ricin is synthesized in castor bean seeds as an endoplasmic reticulum (ER)-targeted precursor. Removal of the signal peptide generates proricin in which the mature A- and B-chains are joined by an intervening propeptide and a 9-residue propeptide persists at the N terminus. The two propeptides are ultimately removed in protein storage vacuoles, where ricin accumulates. Here we have demonstrated that the N-terminal propeptide of proricin acts as a nonspecific spacer to ensure efficient ER import and glycosylation. Indeed, when absent from the N terminus of ricin A-chain, the non-imported material remained tethered to the cytosolic face of the ER membrane, presumably by the signal peptide. This species appeared toxic to ribosomes. The propeptide does not, however, influence catalytic activity per se or the vacuolar targeting of proricin or the rate of retrotranslocation/degradation of A-chain in the cytosol. The likely implications of these findings to the survival of the toxin-producing tissue are discussed.


Adaptation of international guidelines on assessment and management of cancer pain for the Australian context.
To develop clinical practice guidelines for screening, assessing and managing cancer pain in Australian adults. This three-phase project utilized the ADAPTE approach to adapt international cancer pain guidelines for the Australian setting. A Working Party was established to define scope, screen guidelines for adaptation and develop recommendations to support better cancer pain control through screening, assessment, pharmacological and non-pharmacological management, and patient education. Recommendations with limited evidence were referred to Expert Panels for advice before the draft guidelines were opened for public consultation via the Cancer Council Australia Cancer Guidelines Wiki platform in late 2012. All comments were reviewed by the Working Party and the guidelines were revised accordingly. Screening resulted in six international guidelines being included for adaptation - those developed by the Scottish Intercollegiate Guidelines Network (2008), National Health Service Quality Improvement Scotland (2009), National Comprehensive Cancer Network (2012), European Society of Medical Oncology (2011), European Association for Palliative Care (2011, 2012) and National Institute of Clinical Excellence (2012). Guideline adaptation resulted in 55 final recommendations. The guidelines were officially launched in November 2013. International guidelines can be efficiently reconfigured for local contexts using the ADAPTE approach. Availability of the guidelines via the Cancer Council Australia Wiki is intended to promote uptake and enable recommendations to be kept up to date. Resources to support implementation will also be made available via the Wiki if found to be effective by a randomized controlled trial commencing in 2015.


The conditioned place preference is affected by two independent reinforcement processes.
The conditioned place preference method for measuring the affective properties of reinforcing events was studied using treatments of known affective value. The size of the place aversion observed increased with dose when the reinforcer was injections of lithium chloride. The size of the place preference observed increased with concentration when the reinforcer was drinking sucrose solutions. However, when the reinforcer was solutions of saccharin (that were consumed in the same amounts as the sucrose solutions) no place preferences were observed. This finding was explained in terms of the dual reinforcement hypothesis [20] which postulates that although sucrose and saccharin both have positive affective properties (based on their tastes) only sucrose has memory improving properties (based on its post-ingestive action). It was therefore proposed that conditioned place preferences depend on the activation of both affective and memory improving processes. This hypothesis was confirmed by the observation of place preferences with a saccharin solution as the reinforcer when the pairing trials were followed by non-contingent, post-pairing injections of glucose or amphetamine (both of which are known to improve memory). Therefore, behavior in the place preference method depends upon both the affective and the memory improving properties of the reinforcers under test.


[The estimation of the carcinogenic risk in relation to anthropogenic pollution of the atmosphere].
The paper assesses an additional risk of aerogenic exposure in the establishment of a poor epidemiological situation in the context of cancer morbidity, which accounts for 2 cases per 1000 individuals in the most polluted area. The analysis made it possible to reveal the areas showing an increased additional cancer risk by individual nosological entities and to determine the directions of hygienic and epidemiological studies.


Perceived discrimination, patient trust, and adherence to medical recommendations among persons with sickle cell disease.
Adults with sickle cell disease (SCD) report experiencing discriminatory behavior from some healthcare providers. The impact of discrimination on health outcomes in SCD, including adherence to physician recommendations, is not known. Our aim was to evaluate the association between perceived discrimination from healthcare providers and nonadherence to physician recommendations among persons with SCD, and to test the potentially mediating role of patient trust. Patients with SCD (age 15 years and older) participating in the Improving Patient Outcomes with Respect and Trust (IMPORT) Study. Perceived discrimination from healthcare providers and reported adherence to physician recommendations were assessed by patient self-report using items from the 2001 Commonwealth Fund Health Survey. Interpersonal trust in medical professionals was assessed using the short form of the Wake Forest Trust in Medical Professionals instrument. We used a cross-sectional analysis of IMPORT participant data. Multivariable Poisson regression models were used to test the independent association of discrimination with adherence and to test patient trust as a potential mediator. Among 273 SCD patients with complete data on all variables of interest, patients reporting experiences of discrimination in the healthcare system were 53% more likely to also report being nonadherent to physician recommendations. Trust in medical professionals appeared to mediate the discrimination/nonadherence relationship, accounting for 50% of the excess prevalence of nonadherence among those experiencing discrimination. SCD patient perceptions of discriminatory experiences from healthcare providers are associated with greater nonadherence to physician recommendations, and may be a potential factor contributing to disparities in health and health quality among this patient population. Perceived discrimination appears to affect adherence behaviors through the pathway of patient trust. Improving relationships between healthcare providers and SCD patients may improve the trust that SCD patients have in medical professionals, which in turn may improve other outcomes among this underserved patient population.


The clinical effectiveness and cost-effectiveness of peginterferon alfa and ribavirin for the treatment of chronic hepatitis C in children and young people: a systematic review and economic evaluation.
Optimal therapy for children with chronic hepatitis C is unclear. Two treatment regimens are currently licensed in children. To assess the clinical effectiveness and cost-effectiveness of peginterferon alfa-2a (Pegasys®, Roche) and peginterferon alfa-2b [ViraferonPeg®, Merck Sharp & Dohme (MSD)] in combination with ribavirin (RBV), within their licensed indications, for the treatment of chronic hepatitis C virus (HCV) in children and young people aged 3-17 years. Twelve electronic bibliographic databases, including The Cochrane Library, MEDLINE and EMBASE, were searched from inception to November 2012. Bibliographies of retrieved papers, key hepatitis C websites and symposia and manufacturers' submissions to the National Institute for Health and Care Excellence were also searched, and clinical experts were contacted. Systematic reviews of clinical effectiveness and cost-effectiveness were conducted, including studies of health-related quality of life (HRQoL), following standard guidelines to ensure methodological rigour. Clinical effectiveness studies were included if they were in children and young people aged 3-17 years with chronic compensated HCV of any severity, including those with human immunodeficiency virus co-infection and those who were treatment naive or had been previously treated. Eligible interventions were peginterferon alfa-2a or peginterferon alfa-2b, each in combination with RBV, compared against best supportive care (BSC) or against each other, and study designs were randomised controlled trials (RCTs) or non-RCTs, or uncontrolled cohort studies. Outcomes included sustained virological response (SVR) and adverse events. Previously published Markov state-transition economic models of chronic HCV in adults were adapted to estimate the cost-effectiveness of peginterferon alfa-2a and -2b (in combination with RBV), compared with BSC and with one another in children. The model extrapolated the impact of SVR on life expectancy, quality-adjusted life expectancy and lifetime costs. Uncertainty was explored through probabilistic and deterministic sensitivity analyses. Seven studies [two peginterferon alfa-2a and RBV (Copegus®, Roche), and five peginterferon alfa-2b and RBV (Rebetol®, MSD)] were included in the review of clinical effectiveness. Six were single-arm cohort studies and one was a RCT for which only those data for a single arm met the inclusion criteria. Overall, the studies were relatively small and of generally poor quality. SVR rates ranged from 53% to 66% (peginterferon alfa-2a) and 29% to 75% (peginterferon alfa-2b) (49% to 65% if excluding two studies with very small sample sizes). Rates of non-response and relapse were variable and adverse events were generally mild. No studies of cost-effectiveness or HRQoL in children and young people met the inclusion criteria. HRQoL, utilities and costs of treatment were therefore taken from studies of adults with chronic HCV. From this model, peginterferon alfa (-2a or -2b) in combination with RBV was more effective and had lower lifetime costs than BSC. Peginterferon alfa-2a had slightly lower lifetime costs and higher quality-adjusted life-years than peginterferon alfa-2b; therefore, peginterferon alfa-2b was dominated by peginterferon alfa-2a. Results were robust to changes in the sensitivity analyses. There were few good quality studies and parameter data had to be taken from adult studies, which is a limitation of the work. Treatment of children and young people with peginterferon (alfa-2a or -2b) and RBV may be an effective therapy. Results from the independent Markov model suggest that peginterferon (alfa-2a or -2b) in combination with RBV is cost-effective compared with BSC. However, the available evidence is of poor quality. Future research into the impact of these treatments on growth and quality of life in children and young people is recommended. This study is registered as PROSPERO CRD42012002743. The National Institute for Health Research Health Technology Assessment programme.


Molecular forensics in avian conservation: a DNA-based approach for identifying mammalian predators of ground-nesting birds and eggs.
The greater sage-grouse (Centrocercus urophasianus) is a ground-nesting bird from the Northern Rocky Mountains and a species at risk of extinction in in multiple U.S. states and Canada. Herein we report results from a proof of concept that mitochondrial and nuclear DNAs from mammalian predator saliva could be non-invasively collected from depredated greater sage-grouse eggshells and carcasses and used for predator species identification. Molecular forensic approaches have been applied to identify predators from depredated remains as one strategy to better understand predator-prey dynamics and guide management strategies. This can aid conservation efforts by correctly identifying predators most likely to impact threatened and endangered species. DNA isolated from non-invasive samples around nesting sites (e.g. fecal or hair samples) is one method that can increase the success and accuracy of predator species identification when compared to relying on nest remains alone. Predator saliva DNA was collected from depredated eggshells and carcasses using swabs. We sequenced two partial fragments of two mitochondrial genes and obtained microsatellite genotypes using canid specific primers for species and individual identification, respectively. Using this multilocus approach we were able to identify predators, at least down to family, from 11 out of 14 nests (79%) and three out of seven carcasses (47%). Predators detected most frequently were canids (86%), while other taxa included rodents, a striped skunk, and cattle. We attempted to match the genotypes of individual coyotes obtained from eggshells and carcasses with those obtained from fecal samples and coyotes collected in the areas, but no genotype matches were found. Predation is a main cause of nest failure in ground-nesting birds and can impact reproduction and recruitment. To inform predator management for ground-nesting bird conservation, accurate identification of predator species is necessary. Considering predation can have a high impact on recruitment, predation events are very difficult to observe, and predator species are difficult to identify visually from nest remains, molecular approaches that reduce the need to observe or handle animals offer an additional tool to better understand predator-prey dynamics at nesting sites.


Biomechanical assessment of fixation methods for segmental mandible reconstruction with fibula in the polyurethane model.
A variety of plating techniques are employed by microsurgeons to provide rigid fixation for vascularized bone constructs of the mandible. The aim of this study was to biomechanically compare three commonly utilized plating techniques for rigid fixation of fibula bone flaps in reconstructing lateral segmental mandibular defects. Polyurethane mandibles with 3-cm segmental defects were reconstructed using polyurethane fibula models. Three fixation techniques were compared (n = 5 models per group): Group 1 used two 2.0-mm miniplates at each osteotomy site, Group 2 used a single 2.3-mm plate, and Group 3 used a single 2.7-mm plate. Biomechanical testing of maximum force and displacement at failure for each plating technique was assessed and statistical comparison performed. The average displacement for Group 1 was 14.08 ± 1.42 mm, Group 2 was 5.79 ± 0.89 mm, and Group 3 was 6.03 ± 1.59 mm. Group 1 had significantly greater (P < 0.05) displacement when compared with Group 2 and 3. Analysis of variance demonstrated the three groups varied significantly in mean displacement (0 < 0.01). The average force before failure for Group 1 was 616.4 ± 33.83N, Group 2 was 737.8 ± 72.57N, and Group 3 was 681.0 ± 67.98N. Group 2 withstood significantly greater force than Group 1 (P < 0.05), and withstood greater force than Group 3, although the difference was not significant. Analysis of variance showed the three groups varied significantly in mean force at failure (P < 0.05). Reconstruction using a single 2.3-mm plate provided the best rigid fixation for lateral segmental defects of the mandible. © 2016 Wiley Periodicals, Inc. Microsurgery 36:330-333, 2016.


Surveillance of antimicrobial resistance in Neisseria meningitidis strains isolated from invasive cases in Brazil from 2009 to 2016.
To describe the antimicrobial resistance profile of Neisseria meningitidis isolates causing invasive disease in Brazil from 2009 to 2016. Among 3548 N. meningitidis isolates received, 2888 (81.4 %) were analysed for antimicrobial resistance using the broth microdilution technique, as recommended by the Clinical and Laboratory Standards Institute. Isolates were tested for ciprofloxacin, chloramphenicol, ceftriaxone, penicillin G, ampicillin and rifampin. All the isolates tested were susceptible to ceftriaxone, while 953 (33.0 %), 1307 (45.3 %) and 2 (0.07 %) isolates were penicillin G-, ampicillin- and rifampin-intermediate, respectively. Resistance to rifampin, ciprofloxacin and chloramphenicol was shown by three isolates (0.1 %), two isolates (0.07 %) and one (0.03 %) isolate, respectively. Although no isolates were resistant to penicillin G in the period of 2009-2016, our results show an upward trend in minimum inhibitory concentrations (MICs) for this drug as of 2010 (P<0.001). There was no significant difference between different gender and age groups of patients for reduced susceptibility to penicillin G. There was a higher frequency of isolates with reduced susceptibility to penicillin G in the South and Southeast regions (P<0.001). This reduced susceptibility was also associated with serotype 19 inside serogroup B (P<0.001). Despite the decrease in susceptibility to penicillin G and ampicillin observed from 2010, the overall resistance of N. meningitidis isolates to the antimicrobials tested remained uncommon and sporadic, confirming their efficacy for chemoprophylaxis or treatment of invasive meningococcal disease (IMD) in Brazil. Continued surveillance of N. meningitidis antimicrobial susceptibility profiles is important in order to monitor variations in resistance either geographically, over time or in association with emergent clones.


Extraction and Dyeing Techniques of Traditional Vegetable Dyestuffs.
In order to explore the influencing factors of the extraction of vegetable dyes, a research method for the extraction and dyeing of traditional vegetable dyes was proposed. In this paper, the effects of extraction temperature, extraction time, and the ratio of solid to liquid on the extraction effect were discussed through orthogonal experiments. The dyeing process of silk extract was used in this study. The silk fabric was directly dyed with natural dye from peach leaves, compared with the three dyeing methods of premordant, same-bath mordant, and postmordant, and the color fastness to soaping of the dyed fabric was tested. The results show that the color fastness to soaping of leaf-dyed fabrics conforms to the national standard of superior grades, which effectively proves the correctness of the algorithm and model nature and superiority.


Bright green light treatment of depression for older adults [ISRCTN69400161].
Bright white light has been successfully used for the treatment of depression. There is interest in identifying which spectral colors of light are the most efficient in the treatment of depression. It is theorized that green light could decrease the intensity duration of exposure needed. Late Wake Treatment (LWT), sleep deprivation for the last half of one night, is associated with rapid mood improvement which has been sustained by light treatment. Because spectral responsiveness may differ by age, we examined whether green light would provide efficient antidepressant treatment in an elder age group. We contrasted one hour of bright green light (1,200 Lux) and one hour of dim red light placebo (<10 Lux) in a randomized treatment trial with depressed elders. Participants were observed in their homes with mood scales, wrist actigraphy and light monitoring. On the day prior to beginning treatment, the participants self-administered LWT. The protocol was completed by 33 subjects who were 59 to 80 years old. Mood improved on average 23% for all subjects, but there were no significant statistical differences between treatment and placebo groups. There were negligible adverse reactions to the bright green light, which was well tolerated. Bright green light was not shown to have an antidepressant effect in the age group of this study, but a larger trial with brighter green light might be of value.


Familial Risk and Heritability of Cancer Among Twins in Nordic Countries.
Estimates of familial cancer risk from population-based studies are essential components of cancer risk prediction. To estimate familial risk and heritability of cancer types in a large twin cohort. Prospective study of 80,309 monozygotic and 123,382 same-sex dizygotic twin individuals (N = 203,691) within the population-based registers of Denmark, Finland, Norway, and Sweden. Twins were followed up a median of 32 years between 1943 and 2010. There were 50,990 individuals who died of any cause, and 3804 who emigrated and were lost to follow-up. Shared environmental and heritable risk factors among pairs of twins. The main outcome was incident cancer. Time-to-event analyses were used to estimate familial risk (risk of cancer in an individual given a twin's development of cancer) and heritability (proportion of variance in cancer risk due to interindividual genetic differences) with follow-up via cancer registries. Statistical models adjusted for age and follow-up time, and accounted for censoring and competing risk of death. A total of 27,156 incident cancers were diagnosed in 23,980 individuals, translating to a cumulative incidence of 32%. Cancer was diagnosed in both twins among 1383 monozygotic (2766 individuals) and 1933 dizygotic (2866 individuals) pairs. Of these, 38% of monozygotic and 26% of dizygotic pairs were diagnosed with the same cancer type. There was an excess cancer risk in twins whose co-twin was diagnosed with cancer, with estimated cumulative risks that were an absolute 5% (95% CI, 4%-6%) higher in dizygotic (37%; 95% CI, 36%-38%) and an absolute 14% (95% CI, 12%-16%) higher in monozygotic twins (46%; 95% CI, 44%-48%) whose twin also developed cancer compared with the cumulative risk in the overall cohort (32%). For most cancer types, there were significant familial risks and the cumulative risks were higher in monozygotic than dizygotic twins. Heritability of cancer overall was 33% (95% CI, 30%-37%). Significant heritability was observed for the cancer types of skin melanoma (58%; 95% CI, 43%-73%), prostate (57%; 95% CI, 51%-63%), nonmelanoma skin (43%; 95% CI, 26%-59%), ovary (39%; 95% CI, 23%-55%), kidney (38%; 95% CI, 21%-55%), breast (31%; 95% CI, 11%-51%), and corpus uteri (27%; 95% CI, 11%-43%). In this long-term follow-up study among Nordic twins, there was significant excess familial risk for cancer overall and for specific types of cancer, including prostate, melanoma, breast, ovary, and uterus. This information about hereditary risks of cancers may be helpful in patient education and cancer risk counseling.


Periodontal status, salivary immunoglobulin, and microbial counts after short exposure to an isolated environment.
Salivary flow rate, immunoglobulin, and periodontal status were affected during a simulated Skylab mission. The effect is more prominent after long-duration space flights and can persist for several weeks after landing. The objective of this study was to determine the effect of a simulated Mars environment on periodontal status and levels of salivary microorganisms and immunoglobulins in the human oral cavity. Twelve healthy male volunteers were studied before, at 1 and 2 weeks, and after completion of a mission in an isolated, confined simulated Mars environment at the Mars Desert Research Station, USA. We conducted a current stress test, measured salivary immunoglobulin, cortisol, α-amylase, salivary flow rate, and levels of plaque and salivary microbes, and assessed clinical periodontal parameters (probing depth, bleeding on probing, and clinical loss of attachment). Salivary IgG levels and Streptococcus mutans activity were significantly higher at 1 week. Values for clinical periodontal parameters (probing depth, bleeding on probing, and clinical loss of attachment) significantly differed at 1 week. Stress might be caused by the difficulty of the mission rather than the isolated environment, as mission duration was quite short. Periodontal condition might worsen due to poor oral hygiene during the mission. The present findings show that all periodontal conditions and levels of oral bacteria and stress after completion of the simulated Mars mission differed from those at baseline. To verify the relationship between stress status and periodontal health in simulated Mars missions, future studies using larger patient samples and longer follow-up will be required.


Genetic diseases of steroid metabolism.
All major classes of biologically active steroid hormones (progestins, mineralocorticoids, glucocorticoids, and sex steroids) are synthesized from cholesterol through 11 different bioconversions. With the exception of 5 alpha-reductase, all the enzymes mediating these reactions fall into two classes, cytochromes P450 and short-chain dehydrogenases. Cytochromes P450 are heme-containing membrane-bound proteins with molecular weights of approximately 50,000 that utilize molecular oxygen and electrons from NADPH-dependent accessory proteins to hydroxylate substrates. Short-chain dehydrogenases have molecular weights of 30,000-40,000, have tyrosine and lysine residues at the active site, and remove a hydride from the substrate, transferring the electrons of the hydride to NAD+ or NADP+. In most cases, this reaction is reversible so that the dehydrogenase can also function as a reductase under appropriate conditions. Inherited disorders in enzymes required for steroid biosynthesis have varying effects. Defects that prevent cortisol from being synthesized are referred to collectively as congenital adrenal hyperplasia. Because the enzymes required for cortisol biosynthesis in the adrenal cortex are in many cases required for the synthesis of mineralocorticoids and/or sex steroids, these classes of steroids may also not be synthesized normally. Thus, cholesterol desmolase and 3 beta-hydroxysteroid dehydrogenase deficiencies affect synthesis of all classes of steroids in both the adrenals and gonads. Steroid 21-hydroxylase deficiency, the most common cause (> 90% of cases) of congenital adrenal hyperplasia, can affect both mineralocorticoid and glucocorticoid synthesis, but androgen secretion is usually abnormally high due to shunting of accumulated precursors into this pathway. Excessive secretion of androgens and mineralocorticoids occurs in 11 beta-hydroxylase deficiency (the second most frequent form of congenital adrenal hyperplasia). Mineralocorticoid excess is also seen in 17 alpha-hydroxylase deficiency, but in this disorder sex steroid synthesis is defective. All defects that affect estrogen synthesis (deficiencies of cholesterol desmolase, 3 beta-hydroxysteroid dehydrogenase, 17 alpha-hydroxylase, aromatase, and 17 beta-hydroxysteroid dehydrogenase) are very rare, suggesting that the inability to synthesize placental estrogens may adversely affect fetal survival. A number of enzymes are expressed at sites of steroid action and regulate the amount of active steroid available to steroid receptors. Steroid 5 alpha-reductase converts testosterone to the more active dihydrotestosterone. Deficiency of this activity leads to incomplete development of male genitalia; 17 beta-hydroxysteroid dehydrogenase deficiency has similar phenotypic effects.(ABSTRACT TRUNCATED AT 400 WORDS)


Influence of the solvent in the self-assembly and binding properties of [1 + 1] tetra-imine bis-calix[4]pyrrole cages.
We report the self-assembly of shape-persistent [1 + 1] tetra-imine cages 1 based on two different tetra-α aryl-extended calix[4]pyrrole scaffolds in chlorinated solvents and in a 9 : 1 CDCl3 : CD3CN solvent mixture. We show that the use of a bis-N-oxide 4 (4,4'-dipyridyl-N,N'-dioxide) as template is not mandatory to induce the emergence of the cages but has a positive effect on the reaction yield. We use 1H NMR spectroscopy to investigate and characterize the binding properties (kinetic and thermodynamic) of the self-assembled tetra-imine cages 1 with pyridine N-oxide derivatives. The cages form kinetically and thermodynamically stable inclusion complexes with the N-oxides. For the bis-N-oxide 4, we observe the exclusive formation of 1 : 1 complexes independently of the solvent used. In contrast, the pyridine-N-oxide 5 (mono-topic guest) produces inclusion complexes displaying solvent dependent stoichiometry. The bis-N-oxide 4 is too short to bridge the gap between the two endohedral polar binding sites of 1 by establishing eight ideal hydrogen bonding interactions. Nevertheless, the bimolecular 4⊂1 complex results as energetically favored compared to the 52⊂1 ternary counterpart. The inclusion of the N-oxides, 4 and 5, in the tetra-imine cages 1 is significantly faster in chlorinated solvents (minutes) than in the 9 : 1 CDCl3 : CD3CN solvent mixture (hours). We provide an explanation for the similar energy barriers calculated for the formation of the 4⊂1 complex using the two different ternary counterparts 52⊂1 and (CD3CN)2⊂1 as precursors. We propose a mechanism for the in-out guest exchange processes experienced by the tetra-imine cages 1.


Thromboelastography in children with coagulation factor deficiencies.
Hemophilia is traditionally classified according to the levels of the deficient coagulation factor as Severe (<1%), Moderate (1-5%) or Mild (>5%). However, it is well known that the factor activity does not necessarily correspond to the clinical bleeding manifestations. As prophylactic therapy is the best method of prevention of serious complications such as hemophilic arthropathy, a test that may predict the bleeding pattern would be extremely beneficial. Thromboelastography (TEG) uses whole blood to determine clot formation characteristics, such as initiation, propagation as well as strength of the clot, and is now being extensively studied in bleeding and thrombophilia. This study attempted to determine the TEG characteristics in 47 children with moderate hemophilia (MH) and severe hemophilia with (SHI) and without inhibitors (SH) and tried to retrospectively correlate them to the clinical bleeding patterns. TEG showed evidence of faster and better clot formation, as evidenced by a higher maximum thrombin/fibrin generation, in those with mild bleeding manifestations compared to those with severe bleeding tendency, in addition to the expected prolongation in time to formation of clot related to factor deficiency. This may be a potentially useful tool to evaluate the bleeding tendency and determine need for prophylaxis in children with hemophilia.


Context-dependent resistance of freshwater invertebrate communities to drying.
More freshwater ecosystems are drying in response to global change thereby posing serious threat to freshwater biota and functions. The production of desiccation-resistant forms is an important adaptation that helps maintain biodiversity in temporary freshwaters by buffering communities from drying, but its potential to mitigate the negative effects of drying in freshwater ecosystems could vary greatly across regions and ecosystem types. We explored this context dependency by quantifying the potential contribution of desiccation-resistance forms to invertebrate community recovery across levels of regional drying prevalence (defined as the occurrence of drying events in freshwaters in a given region) and ecosystem types (lentic, lotic) in temporary neotropical freshwaters. We first predicted that regional drying prevalence influences the selection of species with desiccation-resistant forms from the regional species pools and thus increases the ability of communities to recover from drying. Second, we predicted lentic freshwaters harbor higher proportions of species with desiccation-resistant forms compared to lotic, in response to contrasted hydrologic connectivity. To test these predictions, we used natural experiments to quantify the contribution of desiccation-resistant forms to benthic invertebrate community recovery in nine intermittent streams and six geographically isolated temporary wetlands from three Bolivian regions differing in drying prevalence. The contribution of desiccation-resistant forms to community recovery was highest where regional drying prevalence was high, suggesting the species pool was adapted to regional disturbance regimes. The contribution of desiccation-resistant forms to community recovery was lower in streams than in wetlands, emphasizing the importance of hydrologic connectivity and associated recolonization processes from in-stream refuges to recovery in lotic systems. In all regions, the majority of functional traits were present in desiccation-resistant taxa indicating this adaptation may help maintain ecosystem functions by buffering communities from the loss of functional traits. Accounting for regional context and hydrologic connectivity in community recovery processes following drying can help refine predictions of freshwater biodiversity response to global change.


[Descriptive study of morbidity and mortality due to asthma at a Health Sector institution].
The objective of this study was to determine the morbidity and mortality rate due to asthma from a Health Institution, which represents the majority of working population. Asthma data were obtained from the National Institute of Informatical, Geography and Statistics (INEGI), Mexican Health Secretary (SS) and Mexican Social Security Institute (IMSS). The underlying cause of death hospitalization or visit were obtained. Asthma was coded according to the International Classification of Disease (ninth revision). Asthma death rate was adjusted by age using direct method. In the IMSS, asthma death rate increased from 3.24/200,000 in 1980 to 12.76/100,000 in 1990. The asthma letality increased from 0.34 in 1980 to 1.23 in 1990. The average length of hospital stay was 3.96 days in 1990; there was noy significant differences by sex the most affected groups were children under 4 years of age and persons older than 65 years of age. In conclusion, from 1980 to 1990, the morbidity and mortality rate due to asthma in IMSS increased. It suggests that future health policy efforts should be focused to reduce the morbidity, mainly in high risk groups.


Integrated microsystems for the in situ genetic detection of dengue virus in whole blood using direct sample preparation and isothermal amplification.
Owing to the frequent outbreak of dengue fever worldwide, a highly sensitive but in situ simple process diagnostic device is required to detect the dengue virus. However, the current immune affinity-based methods have sensitivity issues and nucleic acid-based diagnostic devices have not been suitable for field diagnosis due to the complexity in sample preparation. Here, a simple and fast nucleic acid-based diagnostic tool to directly detect dengue viruses in whole blood is demonstrated using a microbead-assisted direct sample preparation buffer (MB-buffer) and isothermal amplification (loop-mediated isothermal amplification, LAMP). To maximize the performance of the sample preparation process in the microfluidic chip platform, the chemical composition of the sample preparation buffer is simplified and combined with physical tools (heating and bead beating). The entire serial processes consisted of only (1) sample (whole blood) loading, (2) stirring for 90 s, (3) heating at 70 °C for 10 min, and (4) LAMP amplification in the simply designed microfluidic chip cartridge. A single syringe was utilized for sample loading and microfluidic solution transfer. Consequently, dengue viruses were qualitatively detected and discriminated with high sensitivity (LOD: 102 PFU per 200 μL of whole blood) in less than 1 hour without the use of any sophisticated system.


[Whole body PET/CT imaging for detection of metastatic choroidal melanoma].
Choroidal melanoma is the most common primary intraocular cancer in adults. Metastases are most commonly found in the liver and are rapidly fatal despite aggressive therapy. These metastases have been reported in the follow-up to 50 % of uveal melanoma. However, recent reports have shown that liver ultrasonography and liver function tests have low sensitivity in the diagnosis of metastatic uveal melanoma. The study reports on a patient with a history of choroidal melanoma. Preoperative medical evaluation, including CT imaging of the abdomen, complete blood count and liver enzymes proved negative. A PET/CT was requested and showed lesions in the liver and lungs. Whole-body PET/CT revealed melanoma metastases. The PET/CT is a sensitive tool for the detection and localization of hepatic and extrahepatic metastatic choroidal melanoma.


Effect of QT interval-prolonging drugs taken in pregnancy on the neonatal QT interval.
Introduction: Acquired QT interval prolongations due to drug side effects can result in detrimental arrhythmia. Maternal use of placenta-permeable drugs may lead to fetal exposure, thus leading to an increased risk of neonatal QT prolongation and arrhythmia. Objectives: This study aimed to evaluate the influence of maternal QT-prolonging medication on the neonatal QT interval. Methods: In the prospective KUNO-Kids health study, an ongoing population-based birth cohort, we classified maternal medications according to the known risk of QT interval prolongation. Effects on the neonatal QT interval were tested by linear regression analyses, correcting for perinatal confounders (birth weight, gestational age, birth mode, and age at ECG recording). Subgroup analyses were performed for selective serotonin reuptake inhibitors, proton pump inhibitors, and antihistamine dimenhydrinate. Logistic regression analysis was performed using a QTc of 450 ms as the cut-off value. Results: A total of 2,550 pregnant women received a total of 3,990 medications, of which 315 were known to increase the risk of QT prolongation, resulting in 105 (4.1%) neonates exposed in the last month of pregnancy. Overall, the mean age of the neonates at ECG was 1.9 days and the mean QTc (Bazett) was 414 ms. Univariate (regression coefficient -2.62, p = 0.288) and multivariate (regression coefficient -3.55, p = 0.146) regression analyses showed no significant effect of fetal medication exposure on the neonatal QT interval, neither in the overall nor in the subgroup analysis. Logistic regression analysis showed no association of exposure to maternal medication with an increased risk of neonatal QT interval prolongation (OR (odds ratio) 0.34, p = 0.14). Conclusion: The currently used maternal medication results in a relevant number of fetuses exposed to QT interval-prolonging drugs. In our cohort, exposure was found to have no effect on the neonatal QT interval.


Esophageal perforation in an infant: repair with a pleural flap.
Most esophageal perforations regardless of cause, should be treated surgically. Since anatomic features of the esophagus make even small rents difficult to close, a variety of technics have been developed. Flaps of pleura have been used in adults, but to our knowledge this technic has not been previously described in infants. We used a local pleural flap for successful closure of a large esophageal perforation in a 2-week-old infant. Rapid healing occurred without stricture.


Myeloperoxidase deficiency preserves vasomotor function in humans.
Observational studies have suggested a mechanistic link between the leucocyte-derived enzyme myeloperoxidase (MPO) and vasomotor function. Here, we tested whether MPO is systemically affecting vascular tone in humans. A total of 12 135 patients were screened for leucocyte peroxidase activity. We identified 15 individuals with low MPO expression and activity (MPO(low)), who were matched with 30 participants exhibiting normal MPO protein content and activity (control). Nicotine-dependent activation of leucocytes caused attenuation of endothelial nitric oxide (NO) bioavailability in the control group (P < 0.01), but not in MPO(low) individuals (P = 0.12); here the MPO burden of leucocytes correlated with the degree of vasomotor dysfunction (P = 0.008). To directly test the vasoactive properties of free circulating MPO, the enzyme was injected into the left atrium of anaesthetized, open-chest pigs. Myeloperoxidase plasma levels peaked within minutes and rapidly declined thereafter, reflecting vascular binding of MPO. Blood flow in the left anterior descending artery and the internal mammary artery (IMA) as well as myocardial perfusion decreased following MPO injection when compared with albumin-treated animals (P < 0.001). Isolated IMA-rings from animals subjected to MPO revealed markedly diminished relaxation in response to acetylcholine (P < 0.01) and nitroglycerine as opposed to controls (P < 0.001). Myeloperoxidase elicits profound effects on vascular tone of conductance and resistance vessels in vivo. These findings not only call for revisiting the biological functions of leucocytes as systemic and mobile effectors of vascular tone, but also identify MPO as a critical systemic regulator of vasomotion in humans and thus a potential therapeutic target.


Low-dose intravenous heparin infusion in patients with aneurysmal subarachnoid hemorrhage: a preliminary assessment.
Aneurysmal subarachnoid hemorrhage (aSAH) predisposes to delayed neurological deficits, including stroke and cognitive and neuropsychological abnormalities. Heparin is a pleiotropic drug that antagonizes many of the pathophysiological mechanisms implicated in secondary brain injury after aSAH. The authors performed a retrospective analysis in 86 consecutive patients with Fisher Grade 3 aSAH due to rupture of a supratentorial aneurysm who presented within 36 hours and were treated by surgical clipping within 48 hours of their ictus. Forty-three patients were managed postoperatively with a low-dose intravenous heparin infusion (Maryland low-dose intravenous heparin infusion protocol: 8 U/kg/hr progressing over 36 hours to 10 U/kg/hr) beginning 12 hours after surgery and continuing until Day 14 after the ictus. Forty-three control patients received conventional subcutaneous heparin twice daily as deep vein thrombosis prophylaxis. Patients in the 2 groups were balanced in terms of baseline characteristics. In the heparin group, activated partial thromboplastin times were normal to mildly elevated; no clinically significant hemorrhages or instances of heparin-induced thrombocytopenia or deep vein thrombosis were encountered. In the control group, the incidence of clinical vasospasm requiring rescue therapy (induced hypertension, selective intraarterial verapamil, and angioplasty) was 20 (47%) of 43 patients, and 9 (21%) of 43 patients experienced a delayed infarct on CT scanning. In the heparin group, the incidence of clinical vasospasm requiring rescue therapy was 9% (4 of 43, p = 0.0002), and no patient suffered a delayed infarct (p = 0.003). In patients with Fisher Grade 3 aSAH whose aneurysm is secured, postprocedure use of a low-dose intravenous heparin infusion may be safe and beneficial.


Current situation of fungal diseases in Eritrea.
The epidemiology of fungal infections in Eritrea is unknown. Most cases are under-reported due to a lack of diagnostics. This study estimates the burden of serious fungal infections and highlights treatment and diagnostic gaps in the country. All publications related to fungal infections were identified by searches using PubMed/Medline and Google Scholar. Where no data were available, data from neighbouring countries, then sub-Saharan African countries, then other parts of the world were considered for deriving estimates. The Eritrea population was 3,546,427 in 2020. In 2020, HIV/AIDS patients numbered 1400 and TB incidence were 2875. The five-year adult prevalence of asthma (2016-2020) was 41,390, and the total prevalence estimate of chronic obstructive pulmonary disease (COPD) was 308,328. The annual incidence of cryptococcal meningitis and Pneumocystis jirovecii pneumonia in AIDS patients was estimated at 96 and 205 cases. Oesophageal candidiasis incidence is 715 HIV-infected patients. Chronic pulmonary aspergillosis prevalence, including post-tuberculosis cases, was estimated at 1399 (39/100,000). Fungal asthma has a prevalence of 1035 and 1366 in adults. The estimated prevalence of recurrent vulvovaginal candidiasis and tinea capitis is 59,391 and 342,585, respectively. There are no data on candidaemia, but it is estimated at 5/100,000 (177 cases annually). Invasive aspergillosis in leukaemia, lung cancer, COPD and HIV is estimated at 540 cases and fungal keratitis in 514 cases annually. Serious fungal infections are prevalent in Eritrea with approximately 408,164 people (11.5%) affected annually. Studies on fungal diseases to improve diagnosis and treatment are required with the implementation of a national surveillance program.


Neuroethics and Animals: Report and Recommendations From the University of Pennsylvania Animal Research Neuroethics Workshop.
Growing awareness of the ethical implications of neuroscience in the early years of the 21st century led to the emergence of the new academic field of "neuroethics," which studies the ethical implications of developments in the neurosciences. However, despite the acceleration and evolution of neuroscience research on nonhuman animals, the unique ethical issues connected with neuroscience research involving nonhuman animals remain underdiscussed. This is a significant oversight given the central place of animal models in neuroscience. To respond to these concerns, the Center for Neuroscience and Society and the Center for the Interaction of Animals and Society at the University of Pennsylvania hosted a workshop on the "Neuroethics of Animal Research" in Philadelphia, Pennsylvania. At the workshop, expert speakers and attendees discussed ethical issues arising from neuroscience research involving nonhuman animals, including the use of animal models in the study of pain and psychiatric conditions, animal brain-machine interfaces, animal-animal chimeras, cerebral organoids, and the relevance of neuroscience to debates about personhood. This paper highlights important emerging ethical issues based on the discussions at the workshop. This paper includes recommendations for research in the United States from the authors based on the discussions at the workshop, loosely following the format of the 2 Gray Matters reports on neuroethics published by the Presidential Commission for the Study of Bioethical Issues.


Crowding after sudden habitat loss affects demography and social structure in a bat population.
The sudden loss of habitats due to natural or anthropogenic disturbances causes displacement of mobile animals from affected areas to refuge habitats, where large but often transitory concentrations of individuals may occur. While these local density increases have been previously described, the hypothesis that crowding disrupts demographic processes remains largely untested. Here we used the sudden flooding of a river valley by a hydroelectric reservoir as a quasi-experiment to investigate the consequences of crowding on demography, fecundity and social structure in the European free-tailed bat Tadarida teniotis. We monitored bat populations at roosts near and far from the flooded area, before (2013-2014), during (2015) and after (2016) habitat flooding. We assessed population demographic parameters using Capture-Mark-Recapture (CMR) models (3,821 PIT-tagged individuals), and used genetic relatedness among individuals (1,407 individuals genotyped for 14 microsatellite markers) to infer changes in social structure. Habitat loss through flooding was associated with significant but transitory increases in the number of bats using nearby roosts. This may be related to the higher probability of individuals arriving at those roosts during flooding, together with increases in individual local residency through time, particularly among males. Individual apparent survival was highest during flooding and lowest in the following year, while the probability of leaving a roost safe from flooding was higher near the impact area than farther away. Crowding did not negatively affect fecundity, but the arrival of new individuals led to changes in social structure as revealed by lower genetic relatedness between individuals after disturbance at roosts near the flooding area, but not in those farther afield. Our study documents a clear example of crowding effects, suggesting that bats losing roosts due to a hydroelectric reservoir moved to alternative roosts, where local increases in population size and the arrival of new individuals reduced genetic relatedness and apparent survival, but not fecundity. These results support the hypothesis that crowding after habitat loss can disrupt population processes, even though effects may be subtle and short-lived. Also, they point out the need to duly consider crowding effects when assessing and mitigating anthropogenic impacts on animal populations.


Early diagnoses of autism spectrum disorders in Massachusetts birth cohorts, 2001-2005.
We examined trends in autism spectrum disorder diagnoses by age 36 months (early diagnoses) and identified characteristics associated with early diagnoses. Massachusetts birth certificate and early-intervention program data were linked to identify infants born between 2001 and 2005 who were enrolled in early intervention and receiving autism-related services before age 36 months (through December 31, 2008). Trends in early autism spectrum disorders were examined using Cochran-Armitage trend tests. χ² Statistics were used to compare distributions of selected characteristics for children with and without autism spectrum disorders. Multivariate logistic regression analyses were conducted to identify independent predictors of early diagnoses. A total of 3013 children (77.5 per 10,000 study population births) were enrolled in early intervention for autism spectrum disorder by age 36 months. Autism spectrum disorder incidence increased from 56 per 10,000 infants among the 2001 birth cohort to 93 per 10,000 infants in 2005. Infants of mothers younger than 24 years of age, whose primary language was not English or who were foreign-born had lower odds of an early autism spectrum disorder diagnosis. Maternal age older than 30 years was associated with increased odds of an early autism spectrum disorder diagnosis. Odds of early autism spectrum disorders were 4.5 (95% confidence interval: 4.1-5.0) times higher for boys than girls. Early autism spectrum disorder diagnoses are increasing in Massachusetts, reflecting the national trend observed among older children. Linkage of early-intervention program data with population-based vital statistics is valuable for monitoring autism spectrum disorder trends and planning developmental and educational service needs.


[Statistic pitfalls or how should we interprete numbers in the evaluation of a new treatment].
Are cholesterol lowering drugs useful? Do they increase life expectancy? Do third generation oral contraceptives increase the risk of venous thromboembolism? Is there a worldwide decline in semen quality over the last 50 years? Do vitamin supplements improve your child's IQ? Does homeopathy work better than placebo? These questions illustrate some statistical problems and some bias encountered during clinical studies, which can lead to erroneous results. Type I and II errors, surveillance, prescription or publication bias as well as the healthy user effect are described. Problems of regression to the mean, limits of meta-analysis validity and other statistical problems are discussed.


Systematic review of the relative concentrations of noroviruses and fecal indicator bacteria in wastewater: considerations for use in quantitative microbial risk assessment.
Human noroviruses are a leading cause of food- and water-borne disease, which has led to an interest in quantifying norovirus health risks using quantitative microbial risk assessment (QMRA). Given the limited availability of quantitative norovirus data to input to QMRA models, some studies have applied a conversion factor to estimate norovirus exposure based on measured fecal indicator bacteria (FIB) concentrations. We conducted a review of peer-reviewed publications to identify the concentrations of noroviruses and FIB in raw, secondary-treated, and disinfected wastewater. A meta-analysis was performed to determine the ratios of norovirus-FIB pairs in each wastewater matrix and the variables that significantly impact these ratios. Norovirus-to-FIB ratios were found to be significantly impacted by the norovirus genotype, month of sample collection, geographic location, and the extent of wastewater treatment. Additionally, we evaluated the impact of using a FIB-to-virus conversion factor in QMRA and found that the choice of conversion ratio has a great impact on estimated health risks. For example, the use of a conversion ratio previously used in the World Health Organization Guidelines for the Safe Use of Wastewater, Excreta and Greywater predicted health risks that were significantly lower than those estimated with measured norovirus concentrations used as inputs. This work emphasizes the gold standard of using measured pathogen concentrations directly as inputs to exposure assessment in QMRA. While not encouraged, if one must use a FIB-to-virus conversion ratio to estimate norovirus dose, the ratio should be chosen carefully based on the target microorganisms (i.e., strain, genotype, or class), prevalence of disease, and extent of wastewater treatment.