Biomimetic total synthesis of proto-daphniphylline.
Proto-daphniphylline, the imputed biogenetic parent of the Daphniphyllum alkaloids, has been assembled in a biogenetically styled laboratory synthesis in which a pentacyclization process is the fundamental synthetic stratagem. This extraordinary transformation involves the formation of six (r-bonds under the influence of three elementary reagents-potassium hydroxide, anmnonia, and acetic acid. The facility of the process adds credibility to the previous speculation that a similar process is an important step in the biosynthesis of the Daphniphyllum alkaloids.


Implications of climate change for potamodromous fishes.
There is little understanding of how climate change will impact potamodromous freshwater fishes. Since the mid 1970s, a decline in annual rainfall in south-western Australia (a globally recognized biodiversity hotspot) has resulted in the rivers of the region undergoing severe reductions in surface flows (ca. 50%). There is universal agreement amongst Global Climate Models that rainfall will continue to decline in this region. Limited data are available on the movement patterns of the endemic freshwater fishes of south-western Australia or on the relationship between their life histories and hydrology. We used this region as a model to determine how dramatic hydrological change may impact potamodromous freshwater fishes. Migration patterns of fishes in the largest river in south-western Australia were quantified over a 4 year period and were related to a number of key environmental variables including discharge, temperature, pH, conductivity and dissolved oxygen. Most of the endemic freshwater fishes were potamodromous, displaying lateral seasonal spawning migrations from the main channel into tributaries, and there were significant temporal differences in movement patterns between species. Using a model averaging approach, amount of discharge was clearly the best predictor of upstream and downstream movement for most species. Given past and projected reductions in surface flow and groundwater, the findings have major implications for future recruitment rates and population viabilities of potamodromous fishes. Freshwater ecosystems in drying climatic regions can only be managed effectively if such hydro-ecological relationships are considered. Proactive management and addressing existing anthropogenic stressors on aquatic ecosystems associated with the development of surface and groundwater resources and land use is required to increase the resistance and resilience of potamodromous fishes to ongoing flow reductions.


The effect of connectivity on information in neural networks.
We present a mathematical model that quantifies the amount of information exchanged in bi-dimensional networks of nerve cells as a function of network connectivity Q. Upon varying Q over a significant range, we found that, from a certain cell density onwards, 90% of the maximal information transferred I(Q) in a random neuronal network is already reached with just 40% of the total possible connections Q among the cells. As a consequence, the system would not benefit from additional connections in terms of the amount of I(Q), in agreement with the tendency of brains to minimize Q because of its energetic costs. The model may reveal the circuits responsible for neurodegenerative disorders in that neurodegeneration can be regarded as a connective failure affecting information.


Identification of a non-basic domain in the histone H4 N-terminus required for repression of the yeast silent mating loci.
We have shown previously that a stretch of four charged residues (16-19) at the histone H4 N-terminus is involved in repression of the yeast silent mating loci. One of these residues, Lys16, is a site for acetylation, which may prevent repression of the silent mating loci. In this paper we ask whether other sequences in histone H4, possibly in conjunction with H3 residues, are required for repression. We find that even in combination, the other seven acetylatable lysines in H3 and H4 do not function in repression. In contrast, we have found that an adjacent relatively uncharged domain (residues 21-29) is required for repression and that single amino acid insertions and deletions in this region are extremely detrimental. We propose that the basic and non-basic domains together form a DNA (or protein) induced amphipathic alpha-helix required in the formation of a repressive chromatin structure.


Carotid Arterial Hemodynamic in Ischemic Levkoaraiosis Suggests Hypoperfusion Mechanism.
Leukoaraiosis (ILA) is believed to be ischaemic in origin due to its similar location as that of lacunar infarctions and its association with cerebrovascular risk factors. However, its pathophysiology is not well understood. The ischaemic injuries may be a result of increased pulsatility or cerebral hypo-perfusion. We used carotid duplex ultrasound to prove that the underlying mechanism is hypo-perfusion. We compared 55 ILA patients to 44 risk factor-matched controls with normal magnetic resonance imaging (MRI) of the head. ILA diagnosis was based on MRI and was further categorised according to the Fazekas scale. We measured carotid artery blood flow velocity and diameter and calculated carotid blood flow and resistance indexes. Blood flow velocities and blood flows were significantly lower in the ILA group, including diastolic, systolic and mean pressures (p ≤ 0.05). The resistance indices were higher in the ILA group, but the differences were not statistically significant. All the velocities and blood flows showed a decreasing trend with higher Fazekas score, whereas resistance indexes showed an increasing trend. Lower blood flow and higher resistance of carotid arteries are consistent with the hypo-perfusion theory of ILA. Carotid ultrasound could have a diagnostic and prognostic role in ILA patients.


Embolization of pacing electrode fragment into the superolateral vein in the spinal canal causing root compression.
Lead extraction may be complicated by lead fracture and embolization of the lead fragments into the pulmonary circulation or other vasculature causing various signs and symptoms. We present a very unusual case demonstrating embolization of a pacemaker electrode fragment into the superolateral vein in the spinal canal causing root compression. The electrode fragment was removed by spinal surgery, and the patient's symptoms gradually resolved.


Myelin proteins, glycoproteins, and myelin-related enzymes in experimental demyelination of the rabbit optic nerve: sequence of events.
Wallerian degeneration of the rabbit optic nerve was investigated by the technique of retinal ablation which precludes edema, hemorrhage, or macrophage infiltration. After 8 days of degeneration, marked degradation of axons and some myelin abnormalities appeared in the optic nerve, optic chiasma, and optic tract. Myelin lesions were maximal 32 days after retinal destruction. The amount of material stained with a myelin dye decreased drastically between 32 and 90 days after the operation. Biochemical parameters gave the following sequence of events. The concentration of the major periodic acid--Schiff staining glycoproteins was decreased after 2 days, and 6 days later the presence of cholesterol esters was detected in the optic tissue. After 16 days of Wallerian degeneration, the specific activity of 2',3'-cyclic nucleotide 3'-phosphodiesterase not associated with myelin decreased, indicating a possible de-differentiation of oligodendrocytes. Degradation of myelin basic protein became significant at 32 days and the amount of myelin isolated decreased later. The loss of myelin basic protein coincided with a reduction of myelin periodicity as measured in purified fractions by electron microscopy. These results show that secondary myelin destruction in the absence of edema, hemorrhage, or macrophages is a very slow process, and in this situation myelin undergoes a selective and sequential loss of its constituents.


Dissecting Deep Learning Networks-Visualizing Mutual Information.
Deep Learning (DL) networks are recent revolutionary developments in artificial intelligence research. Typical networks are stacked by groups of layers that are further composed of many convolutional kernels or neurons. In network design, many hyper-parameters need to be defined heuristically before training in order to achieve high cross-validation accuracies. However, accuracy evaluation from the output layer alone is not sufficient to specify the roles of the hidden units in associated networks. This results in a significant knowledge gap between DL's wider applications and its limited theoretical understanding. To narrow the knowledge gap, our study explores visualization techniques to illustrate the mutual information (MI) in DL networks. The MI is a theoretical measurement, reflecting the relationship between two sets of random variables even if their relationship is highly non-linear and hidden in high-dimensional data. Our study aims to understand the roles of DL units in classification performance of the networks. Via a series of experiments using several popular DL networks, it shows that the visualization of MI and its change patterns between the input/output with the hidden layers and basic units can facilitate a better understanding of these DL units' roles. Our investigation on network convergence suggests a more objective manner to potentially evaluate DL networks. Furthermore, the visualization provides a useful tool to gain insights into the network performance, and thus to potentially facilitate the design of better network architectures by identifying redundancy and less-effective network units.


Stacked volume holographic gratings for extending the operational wavelength range in LED and solar applications.
A novel stacking procedure is presented for volume phase holographic gratings (VPHGs) recorded in photopolymer material using Corning Willow Glass as a flexible substrate in order to achieve broader angular and spectral selectivity in a diffractive device with high efficiency for solar and LED applications. For the first time to our knowledge, we have shown a device designed for use with a white LED that has the same input and output angles and high efficiency when illuminated by different wavelengths. In this paper, two VPHGs were designed, experimentally recorded, and tested when illuminated at normal incidence. The experimental approach is based on stacking two individual gratings in which the spatial frequency and slant have been tailored to the target wavelength and using real-time on-Bragg monitoring of the gratings in order to control the recorded refractive index modulation, thereby optimizing each grating efficiency for its design wavelength. Lamination of the two gratings together was enabled by using a flexible glass substrate (Corning Willow Glass). Recording conditions were studied in order to minimize the change in diffraction efficiency and peak diffraction angle during lamination and bleaching. The final fabricated stacked device was illuminated by a white light source, and its output was spectrally analyzed. Compared to a single grating, the stacked device demonstrated a twofold increase in angular and wavelength range. The angular and wavelength selectivity curves are in good agreement with the theoretical prediction for this design. This approach could be used to fabricate stacked lenses for white light LED or solar applications.


Mitochondrial toxicity of organic arsenicals: membrane permeability transition pore opening and respiratory dysfunction.
In order to clarify the mitochondrial toxicity mechanism of the organic arsenical MOPIMP (2-methoxy-4-(((4-(oxoarsanyl) phenyl) imino) methyl) phenol), research was carried out at the sub-cell level based on the previous finding that the compound MOPIMP can damage the mitochondria by triggering a burst of ROS. After investigating its influence on isolated mitochondria in vitro, it was demonstrated that a high dose of MOPIMP with short-term exposure can induce mitochondrial swelling, decrease the membrane potential, enhance the permeability of H+ and K+, and induce membrane lipid peroxidation, indicating that it can result in an MPT process in a ROS-mediated and Ca2+-independent manner. Additionally, MPT was also aggravated as a result of impairment of the membrane integrity and membrane fluidity. In addition, short-term incubation between mitochondria and compound MOPIMP promoted the inhibition of respiratory chain complexes I, II, III and IV, as well as damage to the respiration process, which supported the previous finding about the burst of ROS. On the other hand, after long-term exposure by the organic arsenical MOPIMP, mitochondrial metabolic dysfunction was triggered, which was in accordance with perturbation of the respiratory chain complexes as well as the respiration process. This work systematically sheds light on the mitochondrial toxicity mechanism of the organic arsenical MOPIMP, including induction of the MPT process and inhibition of respiratory metabolism, which provides a potential target for organic arsenicals as anti-tumor drugs.


Clinical Relevance of Targeting the Gastrin-Releasing Peptide Receptor, Somatostatin Receptor 2, or Chemokine C-X-C Motif Receptor 4 in Breast Cancer for Imaging and Therapy.
Imaging and therapy using radioligands targeting receptors overexpressed on tumor cells is successfully applied in neuroendocrine tumor patients. Because expression of the gastrin-releasing peptide receptor (GRPR), somatostatin receptor 2 (SSTR2), and chemokine C-X-C motif receptor 4 (CXCR4) has been demonstrated in breast cancer, targeting these receptors using radioligands might offer new imaging and therapeutic opportunities for breast cancer patients. The aim of this study was to correlate messenger RNA (mRNA) expression of GRPR, SSTR2, and CXCR4 with clinicopathologic and biologic factors, and with prognosis and prediction to therapy response, in order to identify specific breast cancer patient groups suited for the application of radioligands targeting these receptors. First, we studied GRPR and SSTR2 expression in 13 clinical breast cancer specimens by in vitro autoradiography and correlated this with corresponding mRNA levels to investigate whether mRNA levels reliably represent cell surface expression. Next, GRPR, SSTR2, and CXCR4 mRNA levels were measured by quantitative reverse transcriptase polymerase chain reaction in 915 primary breast cancer tissues and correlated with known clinicopathologic and biologic factors, disease-free survival, distant metastasis-free survival, and overall survival (DFS, MFS, and OS, respectively). In 224 adjuvant hormonal treatment-naïve estrogen receptor (ER, ESR1)-positive patients who received tamoxifen as first-line therapy for recurrent or metastatic disease, the expression levels of the receptors were correlated with progression-free survival. Our results showed a significant positive correlation between GRPR and SSTR2 expression analyzed by in vitro autoradiography and by quantitative reverse transcriptase polymerase chain reaction (Spearman's rank correlation coefficient [Rs]=0.94, P<0.001, and Rs=0.73, P=0.0042, respectively). Furthermore, high GRPR and SSTR2 mRNA levels were observed more frequently in ESR1-positive specimens, whereas high CXCR4 expression was associated with ESR1-negative specimens. Also, high mRNA expression of CXCR4 was associated with a prolonged DFS, MFS, and OS (multivariate hazard ratio MFS=0.76 [95% confidence interval, 0.64-0.90], P=0.001), whereas high mRNA levels of GRPR were associated with a prolonged progression-free survival after the start of first-line tamoxifen treatment (multivariate hazard ratio=0.68 [95% confidence interval, 0.48-0.97], P=0.031). Our data indicate that imaging and therapy using GRPR or SSTR2 radioligands might especially be beneficial for ESR1-positive breast cancer and CXCR4 radioligands for ESR1-negative breast cancer.


[Productivity of teaching hospitals in the state of São Paulo,Brazil].
Teaching hospitals can be defined as hospitals whose mission is both to provide health care to the population and to train qualified professionals. This represents high costs that need to be balanced by higher productivity and quality. The aim of the present study was to identify the level of productivity in five teaching hospitals in the state of São Paulo, Brazil. For that, we measured indicators such as patient length of stay, patient turnover rate, substitution interval, and number of employees per hospital bed. For a better understanding of the reality of these teaching hospitals, data regarding morbidity and geographic range of influence were also analyzed. Results were compared with the results from a previous study which analyzed private tertiary and secondary hospitals and public secondary hospitals. Productivity was defined as inversely proportional to length of stay, patient turnover rate, substitution interval, and number of employees per hospital bed. We observed that private hospitals have a higher productivity in terms of most indicators, which suggests that these hospitals make a more efficient use of their human, material, and financial resources. Teaching hospitals have a higher length of stay than the other hospitals (median = 7 days); lower turnover rate than private hospitals and similar to public hospitals (median = 3.70 days); higher employee per bed ratio than the private hospitals, and smaller than public hospitals (median = 7 employee/hospital bed). Substitution interval was different for each type of hospital; only public hospitals had a lower substitution interval than teaching hospitals (median = 1.70 days).


Off-axis excitation of the coupled cavity spherical laser interferometer.
The excitation of a concave optical resonator with one planar mirror by a laser source incident normally on the planar mirror is described. The problem is analyzed by describing the resonator fields by both ray tracing and normal mode analysis. The latter is more general, but it is shown that in the appropriate limit the two methods of analysis yield nearly identical solutions. Main emphasis in the analysis is placed on determination of the reflection coefficient, relative mode amplitudes, and field patterns. These are the parameters that are of interest to users of the scanning interferometer and the coupled cavity laser interferometer. The latter is commonly used to diagnose gaseous plasmas.


Effectiveness of Rectal Suction Biopsy in Diagnosing Hirschsprung Disease.
Rectal suction biopsy (RSB) is performed on clinical and radiological suspicion of Hirschsprung disease (HD). Positive histochemical staining for acetylcholinesterase in the lamina propria mucosa and muscularis mucosa together with the absence of ganglion cells in the submucosa confirms HD. A second RSB or a full-thickness biopsy is done in case of doubt. The primary aim of the study was to evaluate the effectiveness of RSB in diagnosing HD. In addition, the results of RSB in preterm born infants were analyzed. This retrospective study included all infants younger than 1 year who had RSB for HD suspicion from January 2004 to April 2014. All histopathological results were reviewed. Sensitivity, specificity, positive, and negative predictive values of the first RSB were calculated. In total, 183 patients (99 boys, mean age 11 weeks) were included. HD was diagnosed in 55 patients (30%). A second biopsy was performed in 12/55 patients (22%), which confirmed HD in 9 patients. One patient did not undergo a second biopsy, but HD was excluded based on clinical follow-up. Thus, HD was eventually excluded in 4 of the 55 patients with a positive first RSB. HD was excluded in the first biopsy in 128 patients (70%). In 19 of those (15%), a second biopsy was done, which led to the diagnosis of HD in 10 patients. Two patients with negative biopsy results were still operated on because of the high clinical suspicion for HD, and the histopathological analysis on the operative specimen confirmed HD. Thus, HD was eventually diagnosed in 12 out of 128 patients. Thus, the sensitivity of the first RSB was 81% and the specificity was 97%. The positive predictive value was 93% and the negative predictive value was 91%. A subanalysis for the 38 preterm born infants was done, with a mean conceptional age at the time of biopsy of 44 weeks (range 34-82). Sensitivity and specificity of RSB were 83 and 97%, respectively, with one false-positive and one false-negative results. The overall sensitivity of the RSB was 81%, with 12 false-negative results, in which cases extra biopsies had been necessary. The specificity was 97%, with four false-positive findings. We found that RSB can also be reliably and safely performed in preterm born infants, which implies there is no reason to postpone a RSB in those infants. New methods, such as calretinin staining, must be considered to achieve better effectiveness of the RSB for diagnosing HD.


Increased muscular challenge in older adults during obstructed gait.
Skeletal muscle strength is known to decline with age. Although lower extremity (LE) muscle strength is critical to maintaining dynamic stability, few studies have investigated lower extremity muscle challenge during activities of daily living. The purpose of this study was to investigate the effects of age and obstructed gait on relative lower extremity muscular challenge, with respect to available joint strength. Fifteen healthy young and fifteen healthy older adults were asked to walk over level ground and step over obstacles. Pre-amplified surface electrodes were used to measure bilateral muscular activation of the gluteus medius (GM), vastus lateralis (VL), and gastrocnemius (GA). Muscle activation signals were normalized to peak magnitudes collected during maximal manual muscle testing (MMT). Normalized magnitudes were analyzed during the double-support phase for gluteus medius and vastus lateralis and during the single-support phase for gastrocnemius. A two-factor ANOVA was used to test for age group effect, with repeated measure of obstacle height. In general, older adults demonstrated greater relative activation levels compared to young adults. Gluteus medius activity was significantly greater in the elderly as compared to young during periods of double-support (weight transfer). Increased obstacle height resulted in greater relative activation in all muscles, confirming the increased challenge to the musculo-skeletal system. While healthy elderly adults were able to successfully negotiate obstacles of different heights during walking, their muscular strength capacity was significantly lower than young adults, resulting in relatively higher muscular demands. The resulting potential for muscular fatigue during locomotion may place individuals at higher risk for trips and/or falls.


A proteomic analysis of human hemodialysis fluid.
The vascular compartment is an easily accessible compartment that provides an opportunity to measure analytes for diagnostic, prognostic, or therapeutic indications. Both serum and plasma have been analyzed extensively by proteomic approaches in an effort to catalog all proteins and polypeptides. Limitations of such approaches in obtaining a comprehensive catalog of proteins include the fact that a handful of proteins constitute over 90% of plasma protein content and that the renal glomeruli filter out proteins and polypeptides that are smaller than 66 kDa from blood. We chose to study hemodialysis fluid because it contains a higher concentration of small proteins and polypeptides and is also simultaneously depleted of the most abundant proteins present in blood. Using gel electrophoresis in combination with LC-MS/MS, we identified 292 proteins of which greater than 70% had not been previously identified from serum or plasma. More than half of the proteins identified from the hemodialysis fluid were smaller than 40 kDa. We also found 50 N-terminally acetylated peptides that allowed us to unambiguously map the N termini of mature forms of the corresponding proteins. Several identified proteins, including cytokines, were only present as predicted transcripts in data bases and thus represent novel proteins. The proteins identified in this study could serve as biomarkers in serum using more sensitive methods such as ELISA-specific antibodies.


Family history of problem drinking is associated with less sensitivity of alcohol demand to a next-day responsibility.
Behavioral economic demand curves measure alcohol consumption as a function of price and may capture clinically relevant individual differences in alcohol-reinforcing efficacy. This study used a novel, behavioral-economic, hypothetical demand-curve paradigm to examine the association between family history of alcohol misuse and individual differences in both alcohol demand and the relative sensitivity of alcohol demand to next-day responsibilities. Participants were 207 college students (47% male, 68.5% White, 27.4% African American, Mage = 19.5 years) who reported at least one heavy drinking episode (5/4 or more drinks on one occasion for a man/woman) in the past month and completed two versions of an alcohol purchase task (APT) that assessed hypothetical alcohol consumption across 17 drink prices. In one APT (standard), students imagined they had no next-day responsibilities, and in the other, they imagined having a 10:00 a.m. test the next day. A series of analyses of covariance indicated that participants with at least one biological parent or grandparent who had misused alcohol reported similar levels of alcohol demand on the standard APT but significantly less sensitivity to the next-day academic responsibility as measured by the percentage of reduction in demand intensity and breakpoint across the no-responsibility and next-day-test conditions. These findings provide initial evidence that APTs might clarify one potential mechanism of risk conferred by family history. Young adult heavy drinkers with a family history of problematic drinking may be less sensitive to next-day responsibilities that might modulate drinking in drinkers without a family history of alcohol problems.


Graphene quantum dots prevent α-synucleinopathy in Parkinson's disease.
Though emerging evidence indicates that the pathogenesis of Parkinson's disease is strongly correlated to the accumulation1,2 and transmission3,4 of α-synuclein (α-syn) aggregates in the midbrain, no anti-aggregation agents have been successful at treating the disease in the clinic. Here, we show that graphene quantum dots (GQDs) inhibit fibrillization of α-syn and interact directly with mature fibrils, triggering their disaggregation. Moreover, GQDs can rescue neuronal death and synaptic loss, reduce Lewy body and Lewy neurite formation, ameliorate mitochondrial dysfunctions, and prevent neuron-to-neuron transmission of α-syn pathology provoked by α-syn preformed fibrils5,6. We observe, in vivo, that GQDs penetrate the blood-brain barrier and protect against dopamine neuron loss induced by α-syn preformed fibrils, Lewy body/Lewy neurite pathology and behavioural deficits.


Sleep and recovery in physicians on night call: a longitudinal field study.
It is well known that physicians' night-call duty may cause impaired performance and adverse effects on subjective health, but there is limited knowledge about effects on sleep duration and recovery time. In recent years occupational stress and impaired well-being among anaesthesiologists have been frequently reported for in the scientific literature. Given their main focus on handling patients with life-threatening conditions, when on call, one might expect sleep and recovery to be negatively affected by work, especially in this specialist group. The aim of the present study was to examine whether a 16-hour night-call schedule allowed for sufficient recovery in anaesthesiologists compared with other physician specialists handling less life-threatening conditions, when on call. Sleep, monitored by actigraphy and Karolinska Sleep Diary/Sleepiness Scale on one night after daytime work, one night call, the following first and second nights post-call, and a Saturday night, was compared between 15 anaesthesiologists and 17 paediatricians and ear, nose, and throat surgeons. Recovery patterns over the days after night call did not differ between groups, but between days. Mean night sleep for all physicians was 3 hours when on call, 7 h both nights post-call and Saturday, and 6 h after daytime work (p < 0.001). Scores for mental fatigue and feeling well rested were poorer post-call, but returned to Sunday morning levels after two nights' sleep. Despite considerable sleep loss during work on night call, and unexpectedly short sleep after ordinary day work, the physicians' self-reports indicate full recovery after two nights' sleep. We conclude that these 16-hour night duties were compatible with a short-term recovery in both physician groups, but the limited sleep duration in general still implies a long-term health concern. These results may contribute to the establishment of safe working hours for night-call duty in physicians and other health-care workers.


Is opium a real risk factor for esophageal cancer or just a methodological artifact? Hospital and neighborhood controls in case-control studies.
Control selection is a major challenge in epidemiologic case-control studies. The aim of our study was to evaluate using hospital versus neighborhood control groups in studying risk factors of esophageal squamous cell carcinoma (ESCC). We compared the results of two different case-control studies of ESCC conducted in the same region by a single research group. Case definition and enrollment were the same in the two studies, but control selection differed. In the first study, we selected two age- and sex-matched controls from inpatient subjects in hospitals, while for the second we selected two age- and sex-matched controls from each subject's neighborhood of residence. We used the test of heterogeneity to compare the results of the two studies. We found no significant differences in exposure data for tobacco-related variables such as cigarette smoking, chewing Nass (a tobacco product) and hookah (water pipe) usage, but the frequency of opium usage was significantly different between hospital and neighborhood controls. Consequently, the inference drawn for the association between ESCC and tobacco use did not differ between the studies, but it did for opium use. In the study using neighborhood controls, opium use was associated with a significantly increased risk of ESCC (adjusted OR 1.77, 95% CI 1.17-2.68), while in the study using hospital controls, this was not the case (OR 1.09, 95% CI 0.63-1.87). Comparing the prevalence of opium consumption in the two control groups and a cohort enrolled from the same geographic area suggested that the neighborhood controls were more representative of the study base population for this exposure. Hospital and neighborhood controls did not lead us to the same conclusion for a major hypothesized risk factor for ESCC in this population. Our results show that control group selection is critical in drawing appropriate conclusions in observational studies.


Intracranial hemorrhage associated with cerebral hyperperfusion syndrome following carotid endarterectomy and carotid artery stenting: retrospective review of 4494 patients.
Intracranial hemorrhage associated with cerebral hyperperfusion syndrome (CHS) following carotid endarterectomy (CEA) or carotid artery stenting (CAS) is a rare but potentially devastating complication. In the present study the authors evaluated 4494 patients with carotid artery stenosis who had undergone CEA or CAS to clarify the clinicopathological features and outcomes of those with CHS and associated intracranial hemorrhage. Patients with postoperative CHS were retrospectively selected, and clinicopathological features and outcomes were studied. Sixty-one patients with CHS (1.4%) were identified, and intracranial hemorrhage developed in 27 of them (0.6%). The onset of CHS peaked on the 6th postoperative day in those who had undergone CEA and within 12 hours in those who had undergone CAS. Results of logistic regression analysis demonstrated that poor postoperative control of blood pressure was significantly associated with the development of intracranial hemorrhage in patients with CHS after CEA (p = 0.0164). Note, however, that none of the tested variables were significantly associated with the development of intracranial hemorrhage in patients with CHS after CAS. Mortality (p = 0.0010) and morbidity (p = 0.0172) rates were significantly higher in patients with intracranial hemorrhage than in those without. Cerebral hyperperfusion syndrome after CEA and CAS occurs with delayed classic and acute presentations, respectively. Although strict control of postoperative blood pressure prevents intracranial hemorrhage in patients with CHS after CEA, there appears to be no relationship between blood pressure control and intracranial hemorrhage in those with CHS after CAS. Finally, the prognosis of CHS in patients with associated intracerebral hemorrhage is poor.


Novel highly-multiplexed AmpliSeq targeted assay for Plasmodium vivax genetic surveillance use cases at multiple geographical scales.
Although the power of genetic surveillance tools has been acknowledged widely, there is an urgent need in malaria endemic countries for feasible and cost-effective tools to implement in national malaria control programs (NMCPs) that can generate evidence to guide malaria control and elimination strategies, especially in the case of Plasmodium vivax. Several genetic surveillance applications ('use cases') have been identified to align research, technology development, and public health efforts, requiring different types of molecular markers. Here we present a new highly-multiplexed deep sequencing assay (Pv AmpliSeq). The assay targets the 33-SNP vivaxGEN-geo panel for country-level classification, and a newly designed 42-SNP within-country barcode for analysis of parasite dynamics in Vietnam and 11 putative drug resistance genes in a highly multiplexed NGS protocol with easy workflow, applicable for many different genetic surveillance use cases. The Pv AmpliSeq assay was validated using: 1) isolates from travelers and migrants in Belgium, and 2) routine collections of the national malaria control program at sentinel sites in Vietnam. The assay targets 229 amplicons and achieved a high depth of coverage (mean 595.7 ± 481) and high accuracy (mean error-rate of 0.013 ± 0.007). P. vivax parasites could be characterized from dried blood spots with a minimum of 5 parasites/µL and 10% of minority-clones. The assay achieved good spatial specificity for between-country prediction of origin using the 33-SNP vivaxGEN-geo panel that targets rare alleles specific for certain countries and regions. A high resolution for within-country diversity in Vietnam was achieved using the designed 42-SNP within-country barcode that targets common alleles (median MAF 0.34, range 0.01-0.49. Many variants were detected in (putative) drug resistance genes, with different predominant haplotypes in the pvmdr1 and pvcrt genes in different provinces in Vietnam. The capacity of the assay for high resolution identity-by-descent (IBD) analysis was demonstrated and identified a high rate of shared ancestry within Gia Lai Province in the Central Highlands of Vietnam, as well as between the coastal province of Binh Thuan and Lam Dong. Our approach performed well in geographically differentiating isolates at multiple spatial scales, detecting variants in putative resistance genes, and can be easily adjusted to suit the needs in other settings in a country or region. We prioritize making this tool available to researchers and NMCPs in endemic countries to increase ownership and ensure data usage for decision-making and malaria policy.


Tryptophan treatment of aggressive psychiatric inpatients.
This double-blind, placebo-controlled study tested the effectiveness of tryptophan (TRP) in the treatment of aggressive psychiatric inpatients. After a baseline observation period of 1 month, patients were randomly assigned to treatment either with TRP (up to 6 g/day) or with placebo. There were 10 subjects in each treatment group. These treatments were administered for 25-35 days, after which the patients were observed for 1 month. Throughout this study, patients were receiving other medications. Injections of antipsychotics and sedatives were administered as needed to control agitated or violent behavior. Blood levels of TRP and other large neutral amino acids were obtained repeatedly, and ratios between TRP and other amino acids were computed. These analyses confirmed significant increases of TRP ratios in TRP-treated patients. TRP treatment had no effect on the number of violent incidents, but it significantly reduced the need for injections of antipsychotics and sedatives. The study thus provided indirect support for beneficial effects of TRP in aggressive psychiatric inpatients.


Impaired small-world efficiency in structural cortical networks in multiple sclerosis associated with white matter lesion load.
White matter tracts, which play a crucial role in the coordination of information flow between different regions of grey matter, are particularly vulnerable to multiple sclerosis. Many studies have shown that the white matter lesions in multiple sclerosis are associated with focal abnormalities of grey matter, but little is known about the alterations in the coordinated patterns of cortical morphology among regions in the disease. Here, we used cortical thickness measurements from structural magnetic resonance imaging to investigate the relationship between the white matter lesion load and the topological efficiency of structural cortical networks in multiple sclerosis. Network efficiency was defined using a 'small-world' network model that quantifies the effectiveness of information transfer within brain networks. In this study, we first classified patients (n = 330) into six subgroups according to their total white matter lesion loads, and identified structural brain networks for each multiple sclerosis group by thresholding the corresponding inter-regional cortical thickness correlation matrix, followed by a network efficiency analysis with graph theoretical approaches. The structural cortical networks in multiple sclerosis demonstrated efficient small-world architecture regardless of the lesion load, an organization that maximizes the information processing at a relatively low wiring cost. However, we found that the overall small-world network efficiency in multiple sclerosis was significantly disrupted in a manner proportional to the extent of total white matter lesions. Moreover, regional efficiency was also significantly decreased in specific brain regions, including the insula and precentral gyrus as well as regions of prefrontal and temporal association cortices. Finally, we showed that the lesions also altered many cortical thickness correlations in the frontal, temporal and parietal lobes. Our results suggest that the white matter lesions in multiple sclerosis might be associated with aberrant neuronal connectivity among widely distributed brain regions, and provide structural (morphological) evidence for the notion of multiple sclerosis as a disconnection syndrome.


Was Aztec and Mixtec turquoise mined in the American Southwest?
Archaeologists have long suggested that prehispanic states in Mesoamerica acquired turquoise through long-distance exchange with groups living in what is now the American Southwest and adjacent parts of northern Mexico. To test this hypothesis, we use lead and strontium isotopic ratios to investigate the geologic provenance of 43 Mesoamerican turquoise artifacts, including 38 mosaic tiles from offerings within the Sacred Precinct of Tenochtitlan (the Mexica or Aztec capital) and 5 tiles associated with Mixteca-style mosaics currently held by the Smithsonian's National Museum of the American Indian. Most of these artifacts have isotopic signatures that differ from turquoise deposits in the American Southwest, but closely match copper deposits and crustal rocks in Mesoamerica. We thus conclude that turquoise used by the Aztecs and Mixtecs likely derives from Mesoamerican sources and was not acquired through long-distance exchange with the Southwest.


Noggin expands neural stem cells in the adult hippocampus.
New neurons are added to the adult hippocampus throughout life and contribute to cognitive functions, including learning and memory. It remains unclear whether ongoing neurogenesis arises from self-renewing neural stem cells (NSCs) or from multipotential progenitor cells that cannot self-renew in the hippocampus. This is primarily based on observations that neural precursors derived from the subventricular zone (SVZ) can be passaged long term, whereas hippocampal subgranular zone (SGZ) precursors are rapidly depleted by passaging. We demonstrate here that high levels of bone morphogenetic protein (BMP) signaling occur in hippocampal but not SVZ precursors in vitro, and blocking BMP signaling with Noggin is sufficient to foster hippocampal cell self-renewal, proliferation, and multipotentiality using single-cell clonal analysis. Moreover, NSC maintenance requires continual Noggin exposure, which implicates BMPs as crucial regulators of NSC aging. In vivo, Noggin is expressed in the adult dentate gyrus and limits BMP signaling in proliferative cells of the SGZ. Transgenic Noggin overexpression in the SGZ increases multiple precursor cell populations but proportionally increases the glial fibrillary acidic protein-positive cell population at the expense of other precursors, suggesting that Noggin acts on NSCs in vivo. To confirm this, we used a dual thymidine analog paradigm to repeatedly label slowly dividing cells over a long duration. We find that small populations of label-retaining cells exist in the SGZ and that Noggin overexpression increases their numbers. Thus, we propose that the adult hippocampus contains a population of NSCs, which can be expanded both in vitro and in vivo by blocking BMP signaling.


DNA cleavage and antitumour activity of platinum(II) and copper(II) compounds derived from 4-methyl-2-N-(2-pyridylmethyl)aminophenol: spectroscopic, electrochemical and biological investigation.
The reaction of the redox-active ligand, Hpyramol (4-methyl-2-N-(2-pyridylmethyl)aminophenol) with K(2)PtCl(4) yields monofunctional square-planar [Pt(pyrimol)Cl], PtL-Cl, which was structurally characterised by single-crystal X-ray diffraction and NMR spectroscopy. This compound unexpectedly cleaves supercoiled double-stranded DNA stoichiometrically and oxidatively, in a non-specific manner without any external reductant added, under physiological conditions. Spectro-electrochemical investigations of PtL-Cl were carried out in comparison with the analogue CuL-Cl as a reference compound. The results support a phenolate oxidation, generating a phenoxyl radical responsible for the ligand-based DNA cleavage property of the title compounds. Time-dependent in vitro cytotoxicity assays were performed with both PtL-Cl and CuL-Cl in various cancer cell lines. The compound CuL-Cl overcomes cisplatin-resistance in ovarian carcinoma and mouse leukaemia cell lines, with additional activity in some other cells. The platinum analogue, PtL-Cl also inhibits cell-proliferation selectively. Additionally, cellular-uptake studies performed for both compounds in ovarian carcinoma cell lines showed that significant amounts of Pt and Cu were accumulated in the A2780 and A2780R cancer cells. The conformational and structural changes induced by PtL-Cl and CuL-Cl on calf thymus DNA and phiX174 supercoiled phage DNA at ambient conditions were followed by electrophoretic mobility assay and circular dichroism spectroscopy. The compounds induce extensive DNA degradation and unwinding, along with formation of a monoadduct at the DNA minor groove. Thus, hybrid effects of metal-centre variation, multiple DNA-binding modes and ligand-based redox activity towards cancer cell-growth inhibition have been demonstrated. Finally, reactions of PtL-Cl with DNA model bases (9-Ethylguanine and 5'-GMP) followed by NMR and MS showed slow binding at Guanine-N7 and for the double stranded self complimentary oligonucleotide d(GTCGAC)(2) in the minor groove.


Comprehensive comparative analysis of strand-specific RNA sequencing methods.
Strand-specific, massively parallel cDNA sequencing (RNA-seq) is a powerful tool for transcript discovery, genome annotation and expression profiling. There are multiple published methods for strand-specific RNA-seq, but no consensus exists as to how to choose between them. Here we developed a comprehensive computational pipeline to compare library quality metrics from any RNA-seq method. Using the well-annotated Saccharomyces cerevisiae transcriptome as a benchmark, we compared seven library-construction protocols, including both published and our own methods. We found marked differences in strand specificity, library complexity, evenness and continuity of coverage, agreement with known annotations and accuracy for expression profiling. Weighing each method's performance and ease, we identified the dUTP second-strand marking and the Illumina RNA ligation methods as the leading protocols, with the former benefitting from the current availability of paired-end sequencing. Our analysis provides a comprehensive benchmark, and our computational pipeline is applicable for assessment of future protocols in other organisms.


Effectiveness of 3 antagonistic bacterial isolates to control Rhizoctonia solani Kühn on lettuce and potato.
Rhizoctonia solani causes yield losses in numerous economically important European crops. To develop a biocontrol strategy, 3 potato-associated ecto- and endophytically living bacterial strains Pseudomonas fluorescens B1, Pseudomonas fluorescens B2, and Serratia plymuthica B4 were evaluated against R. solani in potato and in lettuce. The disease-suppression effect of the 3 biocontrol agents (BCAs) was tested in a growth chamber and in the field. In growth chamber experiments, all 3 BCAs completely or significantly limited the dry mass (DM) losses on lettuce and the disease severity (DS) caused by R. solani on potato sprouts. Strain B1 showed the highest suppression effect (52% on average) on potato. Under field conditions, the DS on both crops, which were bacterized, decreased significantly, and the biomass losses on lettuce decreased significantly as well. The greatest disease-suppression effect on potato was achieved by strain B1 (37%), followed by B2 (33%) and then B4 (31%), whereas the marketable tuber yield increased up to 12% (B1), 6% (B2), and 17% (B4) compared with the pathogen control at higher disease pressure. Furthermore, in all experiments, B1 proved to be the most effective BCA against R. solani. Therefore, this BCA could be a candidate for developing a commercial product against Rhizoctonia diseases. To our knowledge, this is the first report on the high potential of endophytes to be used as a biological control agent against R. solani under field conditions.


Parainfluenza Virus Infection Among Human Immunodeficiency Virus (HIV)-Infected and HIV-Uninfected Children and Adults Hospitalized for Severe Acute Respiratory Illness in South Africa, 2009-2014.
Background. Parainfluenza virus (PIV) is a common cause of acute respiratory tract infections, but little is known about PIV infection in children and adults in Africa, especially in settings where human immunodeficiency virus (HIV) prevalence is high. Methods. We conducted active, prospective sentinel surveillance for children and adults hospitalized with severe acute respiratory illness (SARI) from 2009 to 2014 in South Africa. We enrolled controls (outpatients without febrile or respiratory illness) to calculate the attributable fraction for PIV infection. Respiratory specimens were tested by multiplex real-time reverse-transcription polymerase chain reaction assay for parainfluenza types 1, 2, and 3. Results. Of 18 282 SARI cases enrolled, 1188 (6.5%) tested positive for any PIV type: 230 (19.4%) were type 1; 168 (14.1%) were type 2; 762 (64.1%) were type 3; and 28 (2.4%) had coinfection with 2 PIV types. After adjusting for age, HIV serostatus, and respiratory viral coinfection, the attributable fraction for PIV was 65.6% (95% CI [confidence interval], 47.1-77.7); PIV contributed to SARI among HIV-infected and -uninfected children <5 years of age and among individuals infected with PIV types 1 and 3. The observed overall incidence of PIV-associated SARI was 38 (95% CI, 36-39) cases per 100 000 population and was highest in children <1 year of age (925 [95% CI, 864-989] cases per 100 000 population). Compared with persons without HIV, persons with HIV had an increased relative risk of PIV hospitalization (9.4; 95% CI, 8.5-10.3). Conclusions. Parainfluenza virus causes substantial severe respiratory disease in South Africa among children <5 years of age, especially those that are infected with HIV.


The meaning of living with severe chronic heart failure as narrated by elderly people.
The meaning of elderly patients' experiences of living with chronic heart failure was studied. Narrative interviews were analysed using a phenomenological hermeneutic approach. 'Feeling imprisoned in illness' and 'feeling free despite illness' constituted the themes. These themes were interpreted as describing variations in awareness of the relationship between the self and the body. In theme 'feeling imprisoned in illness' the body's illness and disability hindered the subjects from being themselves. In the theme 'feeling free despite illness' the disabled body was not experienced as limiting, but rather as a part of the self. The patients' understanding of the illness must be interpreted by the caregiver, who also needs to be aware of different modes of communicating feeling about the illness.


High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers.
Correlative light and electron microscopy (CLEM) is an emerging technique which combines functional information provided by fluorescence microscopy (FM) with the high-resolution structural information of electron microscopy (EM). So far, correlative cryo microscopy of frozen-hydrated samples has not reached better than micrometre range accuracy. Here, a method is presented that enables the correlation between fluorescently tagged proteins and electron cryo tomography (cryoET) data with nanometre range precision. Specifically, thin areas of vitrified whole cells are examined by correlative fluorescence cryo microscopy (cryoFM) and cryoET. Novel aspects of the presented cryoCLEM workflow not only include the implementation of two independent electron dense fluorescent markers to improve the precision of the alignment, but also the ability of obtaining an estimate of the correlation accuracy for each individual object of interest. The correlative workflow from plunge-freezing to cryoET is detailed step-by-step for the example of locating fluorescence-labelled adenovirus particles trafficking inside a cell.


Single molecule analysis of Thermus thermophilus SSB protein dynamics on single-stranded DNA.
Single-stranded (ss) DNA binding (SSB) proteins play central roles in DNA replication, recombination and repair in all organisms. We previously showed that Escherichia coli (Eco) SSB, a homotetrameric bacterial SSB, undergoes not only rapid ssDNA-binding mode transitions but also one-dimensional diffusion (or migration) while remaining bound to ssDNA. Whereas the majority of bacterial SSB family members function as homotetramers, dimeric SSB proteins were recently discovered in a distinct bacterial lineage of extremophiles, the Thermus-Deinococcus group. Here we show, using single-molecule fluorescence resonance energy transfer (FRET), that homodimeric bacterial SSB from Thermus thermophilus (Tth) is able to diffuse spontaneously along ssDNA over a wide range of salt concentrations (20-500 mM NaCl), and that TthSSB diffusion can help transiently melt the DNA hairpin structures. Furthermore, we show that two TthSSB molecules undergo transitions among different DNA-binding modes while remaining bound to ssDNA. Our results extend our previous observations on homotetrameric SSBs to homodimeric SSBs, indicating that the dynamic features may be shared among different types of SSB proteins. These dynamic features of SSBs may facilitate SSB redistribution and removal on/from ssDNA, and help recruit other SSB-interacting proteins onto ssDNA for subsequent DNA processing in DNA replication, recombination and repair.


Role of aqueous extract of Azadirachta indica leaves in an experimental model of Alzheimer's disease in rats.
Alzheimer's disease (AD), a progressive neurodegenerative disorder characterized by multiple cognitive deficits, is often accompanied by behavioral disorders and mood changes. Because of the non-availability of proper curative/preventive therapy for AD, the present study was designed to evaluate the possible role of Azadirachta indica in experimental AD in rats. Experimental AD in rats was produced by nucleus basalis magnacellularis lesion with ibotenic acid (IB) and intacerebroventricular administration of colchicine (Col). Different behavioral tests and biochemical analysis were performed to explore the role to A. indica in AD. A. indica exhibited anxiolytic activity in the open field test in Col lesion animals, which was comparable to that of diazepam. In the Elevated plus maze test, A. indica significantly alleviated IB and Col-induced anxiety. IB and Col-induced depression was mitigated by A. indica, and the results were comparable to that of imipramine. In Morris' water maze test, A. indica pre-treatment improved reference memory, working memory and spatial learning, which are at par with the effects of donepezil. Both IB and Col-induced deficits in active avoidance learning and retention of learned behavior were significantly reversed by A. indica. IB and Col-induced increased lipid peroxidase activity was significantly reversed by A. indica (reductions in malondialdehyde level). A. indica stabilized rise in superoxide dismutase and a decreasing trend in acetylcholine-esterase (AChE) activity was seen with IB and Col lesions. A. indica had no effect over the AChE activity. A. indica might be effective in clinical AD by virtue of its cognition enhancement, antidepressant and antianxiety properties.


Sharing ontology between ICD 11 and SNOMED CT will enable seamless re-use and semantic interoperability.
In order to support semantic interoperability in eHealth systems, domain terminologies need to be carefully designed. SNOMED CT and the upcoming ICD-11 represent a new generation of ontology-based terminologies and classifications. The proposed alignment of these two systems and, in consequence, the validity of their cross-utilisation requires a thorough analysis of the intended meaning of their representational units. We present the ICD11 SNOMED CT harmonization process including: a) the clarification of the interpretation of codes in both systems as representing situations rather than conditions, b) the principles proposed for aligning the two systems with the help of a common ontology, c) the high level design of this common ontology, and d) further ontology-driven issues that have arisen in the course of this work.


Asthma self-assessment in a Medicaid population.
Self-assessment of symptoms by patients with chronic conditions is an important element of disease management. A recent study in a commercially-insured population found that patients who received automated telephone calls for asthma self-assessment felt they benefitted from the calls. Few studies have evaluated the effectiveness of disease self-assessment in Medicaid populations. The goals of this study were to: (1) assess the feasibility of asthma self-assessment in a population predominantly insured by Medicaid, (2) study whether adding a gift card incentive increased completion of the self-assessment survey, and (3) evaluate how the self-assessment affected processes and outcomes of care. We studied adults and children aged 4 years and older who were insured by a Medicaid-focused managed care organization (MCO) in a pre- and post-intervention study. During the pre-incentive period, patients with computerized utilization data that met specific criteria for problematic asthma control were mailed the Asthma Control Test (ACT), a self-assessment survey, and asked to return it to the MCO. During the intervention period, patients were offered a $20 gift card for returning the completed ACT to the MCO. To evaluate clinical outcomes, we used computerized claims data to assess the number of hospitalization visits and emergency department visits experienced in the 3 months after receiving the ACT. To evaluate whether the self-management intervention improved processes of care, we conducted telephone interviews with patients who returned or did not return the ACT by mail. During the pre-incentive period, 1183 patients were identified as having problems with asthma control; 25 (2.0%) of these returned the ACT to the MCO. In contrast, during the incentive period, 1612 patients were identified as having problems with asthma control and 87 (5.4%) of these returned the ACT to the MCO (p < 0.0001). Of all 95 ACTs that were returned, 87% had a score of 19 or less, which suggested poor asthma control.During the 3 months after they received the ACT, patients who completed it had similar numbers of outpatient visits, emergency department visits, and hospitalizations for asthma as patients who did not complete the ACT. We completed interviews with 95 patients, including 28 who had completed the ACT and 67 who had not. Based on an ACT administered at the time of the interview, patients who had previously returned the ACT to the MCO had asthma control similar to those who had not (mean scores of 14.2 vs. 14.6, p = 0.70). Patients had similar rates of contacting their providers within the past 2 months whether they had completed the mailed ACT or not (71% vs. 76%, p = 0.57). Mailing asthma self-assessment surveys to patients with poorly controlled asthma was not associated with better asthma-associated outcomes or processes of care in the Medicaid population studied. Adding a gift card incentive did not meaningfully increase response rates. Asthma disease management programs for Medicaid populations will most likely need to involve alternative strategies for engaging patients and their providers in managing their conditions.


Characterization and genetic dissection of resistance to spotted alfalfa aphid (Therioaphis trifolii) in Medicago truncatula.
Aphids cause significant yield losses in agricultural crops worldwide. Medicago truncatula, a model legume, cultivated pasture species in Australia and close relative of alfalfa (Medicago sativa), was used to study the defence response against Therioaphis trifolii f. maculate [spotted alfalfa aphid (SAA)]. Aphid performance and plant damage were compared among three accessions. A20 is highly susceptible, A17 has moderate resistance, and Jester is strongly resistant. Subsequent analyses using A17 and A20, reciprocal F1s and an A17×A20 recombinant inbred line (RIL) population revealed that this moderate resistance is phloem mediated and involves antibiosis and tolerance but not antixenosis. Electrical penetration graph analysis also identified a novel waveform termed extended potential drop, which occurred following SAA infestation of M. truncatula. Genetic dissection using the RIL population revealed three quantitative trait loci on chromosomes 3, 6, and 7 involved in distinct modes of aphid defence including antibiosis and tolerance. An antibiosis locus resides on linkage group 3 (LG3) and is derived from A17, whereas a plant tolerance and antibiosis locus resides on LG6 and is derived from A20, which exhibits strong temporary tolerance. The loci identified reside in regions harbouring classical resistance genes, and introgression of these loci in current medic cultivars may help provide durable resistance to SAA, while elucidation of their molecular mechanisms may provide valuable insight into other aphid-plant interactions.


Improving glycemic control and cardiometabolic risk through integrated treatment plans.
Type 2 diabetes mellitus (T2DM) is associated with an increased risk of cardiovascular complications. This increased risk is present even before the diagnosis of diabetes is made. Therefore, it is imperative that physicians are aware of risk factors for diabetes and actively screen high-risk patients and diagnose the condition as early as possible. Patients with prediabetes must be counseled on interventions to prevent the progression to diabetes. Integrated treatment plans that address all aspects of disease must be implemented. Patients with diabetes should be examined for dyslipidemia, overweight or obesity, and hypertension. Treatment for patients with T2DM should include lifestyle interventions and should address any other comorbidities for which the patient may be at risk.


The "minimizing antibiotic resistance in Colorado" project: impact of patient education in improving antibiotic use in private office practices.
To assess the marginal impact of patient education on antibiotic prescribing to children with pharyngitis and adults with acute bronchitis in private office practices. Antibiotic prescription rates based on claims data from four managed care organizations in Colorado during baseline (winter 2000) and study (winter 2001) periods. A nonrandomized controlled trial of a household and office-based patient educational intervention was performed. During both periods, Colorado physicians were mailed antibiotic prescribing profiles and practices guidelines as part of an ongoing quality improvement program. Intervention practices (n=7) were compared with local and distant control practices. Office visits were extracted by managed care organizations using International Classification of Diseases-9-Clinical Modification codes for acute respiratory tract infections, and merged with pharmacy claims data based on visit and dispensing dates coinciding within 2 days. Adjusted antibiotic prescription rates during baseline and study periods increased from 38 to 39 percent for pediatric pharyngitis at the distant control practices, and decreased from 39 to 37 percent at the local control practices, and from 34 to 30 percent at the intervention practices (p=.18 compared with distant control practices). Adjusted antibiotic prescription rates decreased from 50 to 44 percent for adult bronchitis at the distant control practices, from 55 to 45 percent at the local control practices, and from 60 to 36 percent at the intervention practices (p<.002 and p=.006 compared with distant and local control practices, respectively). In office practices, there appears to be little room for improvement in antibiotic prescription rates for children with pharyngitis. In contrast, patient education helps reduce antibiotic use for adults with acute bronchitis beyond that achieved by physician-directed efforts.


Breast carcinomatous tumoral emboli can result from encircling lymphovasculogenesis rather than lymphovascular invasion.
The canonical view of the origin of tumor lymphovascular emboli is that they usually originate from lymphovascular invasion as part of a multistep metastatic process. Recent experimental evidence has suggested that metastasis can occur earlier than previously thought and we found evidence that tumor emboli formation can result from the short-circuiting step of encircling lymphovasculogenesis. Experimentally, we used a xenograft of human inflammatory breast cancer (MARY-X), a model that exhibited florid tumor emboli, to generate tumoral spheroids in vitro. In observational studies, we chose human breast carcinoma cases where there appeared to be a possible transition of in situ carcinoma to lymphovascular emboli without intervening stromal invasion. These cases were studied by morphometry as well as IHC with tumor proliferation (Ki-67) and adhesion (E-cadherin) markers, myoepithelial (p63), as well as endothelial (podoplanin [D2-40], CD31, VEGFR-3, Prox-1) markers. Unlabelled spheroids coinjected with either GFP or RFP-human myoepithelial cells or murine embryonal fibroblasts (MEFs) gave rise to tumors which exhibited GFP/RFP immunoreactivity within the cells lining the emboli-containing lymphovascular channels. In vitro studies demonstrated that the tumoral spheroids induced endothelial differentiation of cocultured myoepithelial cells and MEFs, measured by real time PCR and immunofluorescence. In humans, the in situ clusters exhibited similar proliferation, E-cadherin immunoreactivity and size as the tumor emboli (p =.5), suggesting the possibility that the latter originated from the former. The in situclusters exhibited a loss (50%-100%) of p63 myoepithelial immunoreactivity but not E-cadherin epithelial immunoreactivity. The tumor emboli were mainly present within lymphatic channels whose dual p63/CD31, p63/D2-40 and p63/VEGFR-3 and overall weak patterns of D2-40/CD31/VEGFR-3 immunoreactivities suggested that they represented immature and newly created vasculature derived from originally myoepithelial-lined ducts. Collectively both experimental as well as observational studies suggested the possibility that these breast cancer emboli resulted from encircling lymphovasculogenesis rather than conventional lymphovascular invasion.


Tub bathing versus traditional sponge bathing for the newborn.
To compare selected effects of tub bathing versus traditional sponge bathing in healthy, term newborns and their mothers' ratings of pleasure and confidence with the bath. Randomized controlled study. The maternity unit of an eastern Canadian hospital. One hundred two mother-baby pairs were randomly assigned to an experimental tub bath or a sponge bath control group. Fifty-one newborns were tub bathed and 51 sponge bathed according to the study protocols for their initial and one additional bath. (a) Newborn temperature stability was assessed by recording axillary temperatures pre- and postbath, (b) umbilical cord healing was identified by daily observations and infection control surveillance, (c) infant contentment was quantified by applying the Brazelton Neonatal Behavioral Assessment Scale, and (d) maternal pleasure with the bath and confidence with bathing at discharge were self-rated on a 5-point scale. Tub-bathed babies experienced significantly less temperature loss (t = 4.79, p = .00) and were significantly more content (t = -6.48, p = .00) than were those who were sponge bathed. No differences in cord healing scores were found. Mothers of tub bathed babies rated their pleasure with the bath significantly higher than did mothers of sponge bathed babies (t = 4.15, p = .00). No differences in maternal confidence were noted. Tub bathing is a safe and pleasurable alternative to sponge bathing in healthy, term newborns.


Bone morphogenetic proteins inhibit the tumorigenic potential of human brain tumour-initiating cells.
Transformed, oncogenic precursors, possessing both defining neural-stem-cell properties and the ability to initiate intracerebral tumours, have been identified in human brain cancers. Here we report that bone morphogenetic proteins (BMPs), amongst which BMP4 elicits the strongest effect, trigger a significant reduction in the stem-like, tumour-initiating precursors of human glioblastomas (GBMs). Transient in vitro exposure to BMP4 abolishes the capacity of transplanted GBM cells to establish intracerebral GBMs. Most importantly, in vivo delivery of BMP4 effectively blocks the tumour growth and associated mortality that occur in 100% of mice after intracerebral grafting of human GBM cells. We demonstrate that BMPs activate their cognate receptors (BMPRs) and trigger the Smad signalling cascade in cells isolated from human glioblastomas (GBMs). This is followed by a reduction in proliferation, and increased expression of markers of neural differentiation, with no effect on cell viability. The concomitant reduction in clonogenic ability, in the size of the CD133+ population and in the growth kinetics of GBM cells indicates that BMP4 reduces the tumour-initiating cell pool of GBMs. These findings show that the BMP-BMPR signalling system--which controls the activity of normal brain stem cells--may also act as a key inhibitory regulator of tumour-initiating, stem-like cells from GBMs and the results also identify BMP4 as a novel, non-cytotoxic therapeutic effector, which may be used to prevent growth and recurrence of GBMs in humans.


Galloyl-RGD as a new cosmetic ingredient.
The cosmetics market has rapidly increased over the last years. For example, in 2011 it reached 242.8 billion US dollars, which was a 3.9% increase compared to 2010. There have been many recent trials aimed at finding the functional ingredients for new cosmetics. Gallic acid is a phytochemical derived from various herbs, and has anti-fungal, anti-viral, and antioxidant properties. Although phytochemicals are useful as cosmetic ingredients, they have a number of drawbacks, such as thermal stability, residence time in the skin, and permeability through the dermal layer. To overcome these problems, we considered conjugation of gallic acid with a peptide. We synthesized galloyl-RGD, which represents a conjugate of gallic acid and the peptide RGD, purified it by HPLC and characterized by MALDI-TOF with the aim of using it as a new cosmetic ingredient. Thermal stability of galloyl-RGD was tested at alternating temperatures (consecutive 4°C, 20°C, or 40°C for 8 h each) on days 2, 21, 41, and 61. Galloyl-RGD was relatively safe to HaCaT keratinocytes, as their viability after 48 h incubation with 500 ppm galloyl-RGD was 93.53%. In the group treated with 50 ppm galloyl-RGD, 85.0% of free radicals were removed, whereas 1000 ppm galloyl-RGD suppressed not only L-DOPA formation (43.8%) but also L-DOPA oxidation (54.4%). Galloyl-RGD is a promising candidate for a cosmetic ingredient.


Correlation of the aqueous humor total antioxidant capacity, total oxidant status, and levels of IL-6 and VEGF with diabetic retinopathy status.
We aimed to compare the aqueous humor total oxidant status, total antioxidant capacity, and levels of interleukin-6 and vascular endothelial growth factor between patients with diabetic retinopathy and controls and to correlate these levels with the DR status. Patients who underwent cataract surgery were enrolled. The first group (control group) comprised patients without diabetes; the second group comprised diabetic patients without retinopathy; the third group comprised patients with nonproliferative diabetic retinopathy; and the fourth group comprised patients with proliferative diabetic retinopathy. All patients underwent full ophthalmologic examination before cataract surgery. Prior to surgery, samples of aqueous humor sampling were obtained and stored at -80 °C. Total antioxidant capacity, total oxidant status, and levels interleukin-6 and vascular endothelial growth factor were investigated in these samples and correlated with diabetic retinopathy status. This study analyzed 86 pairs of eyes of 86 patients. All groups were statistically similar in age and sex, but the total antioxidant capacity was lowest in patients with proliferative diabetic retinopathy. Moreover, the total oxidant status and levels of vascular endothelial growth factor and interleukin-6 were found to slightly increase according to the retinopathy status. Oxidative stress, interleukin-6, and vascular endothelial growth factor in the aqueous humor seem to play important roles in the pathogenesis of diabetic retinopathy, especially in the proliferative type.


Human La antigen is required for the hepatitis C virus internal ribosome entry site-mediated translation.
The 5'-noncoding region (5'-NCR) of the hepatitis C virus (HCV) RNA genome serves as an internal ribosome entry site (IRES) and mediates translation initiation in a cap-independent manner. Previously, we reported the interaction between La antigen and the HCV IRES, which appeared to occur in the context of initiator AUG. It was further shown that HCV IRES-mediated translation was stimulated in the presence of human La antigen. In this study, we have defined the cis- and trans-acting elements responsible for La-5'-NCR interactions and established the dependence of the HCV IRES efficiency on cellular La antigen. During the La-IRES interaction, initiator AUG but not the neighboring codons was found to be the direct target of La binding. The C terminus effector domain-dependent modulation of La binding to the HCV IRES is demonstrated by deletion and substitution mutagenesis of the protein. An RNA systematic evolution of ligands by exponential enrichment (SELEX), generated against La protein that selectively binds La in HeLa lysates and competes for the protein binding to the 5'-NCR, was used to demonstrate the requirement of La for the HCV IRES function in the context of mono- and dicistronic mRNAs. Sequestration of La antigen by the RNA SELEX in HeLa translation lysates blocked the HCV and poliovirus IRES-mediated translation in vitro. The functional requirement of La protein for the HCV IRES activity was further established in a liver-derived cell line and in an add-back experiment in which the inhibited IRES was rescued by recombinant human La. These results strongly argue for the novel role of La protein during selection of the initiator AUG and its participation during internal initiation of translation of the HCV RNA genome.


The presence of prostate-specific antigen-related genes in primates and the expression of recombinant human prostate-specific antigen in a transfected murine cell line.
Human prostate-specific antigen (PSA) has been shown as an aid in the early detection of prostate cancer (W. J. Catalona et al., J. Am. Med. Assoc., 270: 948-954, 1993) and was approved in 1994 by the Food and Drug Administration for early detection of prostate cancer. Immunotherapies directed against PSA have been suggested in patients with metastatic prostate cancer. One of the essential questions is to define which nonhuman species express PSA for experimental studies. Using Southern blot analyses, genes related to human PSA have been detected in several nonhuman primate species, including chimpanzee, orangutan, gorilla, macaque, and rhesus monkey, but not in other mammalian species, including rabbit, cow, pig, dog, rat, or mouse. Immunohistochemical staining with anti-human PSA antisera detected strong staining in both human and monkey prostatic epithelial cells with no reactivity to rat prostate cells. Because the PSA gene is not present in the murine genome, a matched set of murine cell lines has been developed that may be useful to study the biochemical functions of PSA and as an experimental target for PSA-directed immunotherapy. To establish such cell lines, a C57BL/6 murine colon adenocarcinoma cell line, MC-38, was transfected with a retroviral vector containing cDNA encoding the human PSA gene. Genetic analysis of a PSA-secreting clone, PSA/MC-38, demonstrated that the PSA gene had been stably integrated into the MC-38 genome. The PSA/MC-38 cell line was found to secrete PSA into tissue culture medium, producing a protein of approximately M(r) 30,000. In vivo, PSA/MC-38 grew as a s.c. tumor in male and female mice. PSA/MC-38 tumors grew more rapidly in athymic mice than in syngeneic C57BL/6 mice, and in both mouse strains, the PSA/MC-38 tumors grew more slowly than control vector-transduced tumors. PSA was detected in the serum and tumors of PSA/MC-38 tumor-bearing mice. It is proposed that PSA/MC-38 cells may be used as a murine tumor model to test potential therapeutic vaccines and other experimental therapies directed against PSA.


Binding of NF-kappaB p65 subunit to the promoter elements is involved in LPS-induced transactivation of miRNA genes in human biliary epithelial cells.
The majority of human miRNA genes is transcribed by polymerase II and can be classified as class II genes similar to protein-coding genes. Whereas current research on miRNAs has focused on the physiological and pathological functions, the molecular mechanisms underlying their transcriptional regulation are largely unknown. We recently reported that lipopolysaccharide (LPS) alters mature miRNA expression profile in human biliary epithelial cells. In this study, we tested the role of transcription factor NF-kappaB in LPS-induced transcription of select miRNA genes. Of the majority of LPS-up-regulated mature miRNAs in cultured human biliary epithelial cells, potential NF-kappaB binding sites were identified in the putative promoter elements of their corresponding genes. Inhibition of NF-kappaB activation by SC-514, an IKK2 inhibitor, blocked LPS-induced up-regulation of a subset of pri-miRNAs, including pri-miR-17-92, pri-miR-125b-1, pri-miR-21, pri-miR-23b-27b-24-1, pri-miR-30b, pri-miR-130a and pri-miR-29a. Moreover, direct binding of NF-kappaB p65 subunit to the promoter elements of mir-17-92, mir-125b-1, mir-21, mir-23b-27b-24-1, mir-30b and mir-130a genes was identified by chromatin immunoprecipitation analysis and confirmed by the luciferase reporter assay. Thus, a subset of miRNA genes is regulated in human biliary epithelial cells through NF-kappaB activation induced by LPS, suggesting a role of the NF-kappaB pathway in the transcriptional regulation of miRNA genes.


STUDIES ON THE SENSITIZATION OF ANIMALS WITH SIMPLE CHEMICAL COMPOUNDS.
Experiments on the sensitization of guinea pigs with simple chemical compounds are described. Positive effects were obtained by the administration of small quantities, namely fractions of milligrams, with 1:2:4 chlorodinitrobenzene, p-nitrosodimethylaniline, 1:2:4 trinitrobenzene, picryl chloride, four dichlorodinitrobenzenes, and a number of other aromatic compounds. Several substances chemically similar to those enumerated gave negative results. The first named compound is known to produce hypersensitiveness in human beings, a large number of cases having been observed in factory workers. The mechanism of these effects is discussed.


Flying over uneven moving terrain based on optic-flow cues without any need for reference frames or accelerometers.
Two bio-inspired guidance principles involving no reference frame are presented here and were implemented in a rotorcraft, which was equipped with panoramic optic flow (OF) sensors but (as in flying insects) no accelerometer. To test these two guidance principles, we built a tethered tandem rotorcraft called BeeRotor (80 grams), which was tested flying along a high-roofed tunnel. The aerial robot adjusts its pitch and hence its speed, hugs the ground and lands safely without any need for an inertial reference frame. The rotorcraft's altitude and forward speed are adjusted via two OF regulators piloting the lift and the pitch angle on the basis of the common-mode and differential rotor speeds, respectively. The robot equipped with two wide-field OF sensors was tested in order to assess the performances of the following two systems of guidance involving no inertial reference frame: (i) a system with a fixed eye orientation based on the curved artificial compound eye (CurvACE) sensor, and (ii) an active system of reorientation based on a quasi-panoramic eye which constantly realigns its gaze, keeping it parallel to the nearest surface followed. Safe automatic terrain following and landing were obtained with CurvACE under dim light to daylight conditions and the active eye-reorientation system over rugged, changing terrain, without any need for an inertial reference frame.


Effects of spatial and temporal context on color categories and color constancy.
Color constancy is the ability to assign a constant color to an object independent of changes in illumination. Color constancy is achieved by taking context information into account. Previous approaches that have used matching paradigms to quantify color constancy found degrees of constancy between 20% and 80%. Here, we studied color constancy in a color-naming task under different conditions of surround illumination and patch size. Observers categorized more than 400 patches for each illumination condition. This allows one to overcome inherent limitations in color naming and to study the changes in color categories under illumination changes. When small central test patches with a full context illumination were categorized, observers followed the illumination shift almost completely, showing a high degree of constancy (99%). Reducing the available context information or increasing the patch size decreased the degree of constancy to about 50%. Moderate degrees of constancy (66%) occurred even when the test patches were never viewed simultaneously but only in temporal alternation with the illumination. Boundaries between color categories were largely stable within and across observers under neutral illumination. Under changing illumination, there were small but systematic variations in the color category boundaries. Color category boundaries tended to rotate away from the illumination color. This variation was largest under full context conditions where highest degrees of color constancy were obtained.


MiR-34b is associated with clinical outcome in triple-negative breast cancer patients.
Breast cancer is the most common malignancy with the highest incidence rates among women worldwide. Triple-negative breast cancer (TNBC) represents the major phenotype of basal-like molecular subtype of breast cancer, characterized by higher incidence in young women and a very poor prognosis. MicroRNAs (miRNAs) are small non-coding RNAs playing significant role in the pathogenesis of many cancers including breast cancer. Therefore, miRNAs are also potential prognostic and/or predictive biomarkers in triple-negative breast cancer patients. Thirty-nine TNBC patients with available formalin-fixed paraffin-embedded (FFPE) tissues were enrolled in the study. MiR-34a, miR-34b, and miR-34c were analyzed using qRT-PCR and correlated to clinico-pathological features of TNBC patients. Expression levels of miR-34b significantly correlate with disease free survival (DFS) (p = 0.0020, log-rank test) and overall survival (OS) (p = 0.0008, log-rank test) of TNBC patients. No other significant associations between miR-34a, miR-34b, and miR-34c with available clinical pathological data were observed. MiR-34b expression negatively correlates with disease free survival and overall survival in TNBC patients. Thus, miR-34b may present a new promising prognostic biomarker in TNBC patients, but independent validations are necessary.


Reactive oxygen species-dependent apoptosis induction by water extract of Citrus unshiu peel in MDA-MB-231 human breast carcinoma cells.
Although several recent studies have reported the anti-cancer effects of extracts or components of Citrus unshiu peel, which has been used for various purposes in traditional medicine, the molecular mechanisms for their effects remain unclear. In the present study, the anti-cancer activity of a water-soluble extract of C. unshiu peel (WECU) in MDA-MB-231 human breast carcinoma cells at the level of apoptosis induction was investigated. Cytotoxicity was evaluated using the MTT assay. Apoptosis was detected using DAPI staining and flow cytometry analyses. Mitochondrial membrane potential, reactive oxygen species (ROS) assay, caspase activity and Western blotting were used to confirm the basis of apoptosis. The results indicated that WECU-induced apoptosis was related to the activation of caspase-8, and -9, representative initiator caspases of extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3 accompanied by proteolytic degradation of poly(ADP-ribose) polymerase and down-regulation of the inhibitors of apoptosis protein family members. WECU also increased the pro-apoptotic BAX to anti-apoptotic BCL-2 ratio, loss of mitochondrial membrane potential and cytochrome c release from mitochondria to cytoplasm. Furthermore, WECU provoked the generation of ROS, but the reduction of cell viability and induction of apoptosis by WECU were prevented when ROS production was blocked by antioxidant N-acetyl cysteine. These results suggest that WECU suppressed proliferation of MDA-MB-231 cells by activating extrinsic and intrinsic apoptosis pathways in a ROS-dependent manner.


Practice Guidelines for Enteral Nutrition Management in Dysglycemic Critically Ill Patients: A Relook for Indian Scenario.
Intensive-care practices and settings differ for India in comparison to other countries. While guidelines are available to direct the use of enteral nutrition (EN), there are no recommendations specific to nutritional management of EN in dysglycemic patients, specific to patients in Indian critical care settings. Advisory board meetings were arranged to develop the practice guidelines specific to the Indian context, for the use of EN in dysglycemic critically ill patients and to overcome challenges in this field. Two advisory board meetings were organized to review various existing guidelines, meta-analyses, randomized controlled trials (RCTs), controlled trials and review articles, for their contextual relevance and strength. Three rounds of Delphi voting were done to arrive at consensus on certain recommendations. A systematic grading of practice guidelines by the advisory board was done based on strength of the consensus voting and reviewed supporting evidences. Based on the literature review, the recommendations for developing the practice guidelines were made as per the grading criteria agreed upon by the advisory board. The recommendations were to address challenges regarding prediction and assessment of dysglycemia (DG), acceptable glycemic targets in such settings, general nutritional aspects pertaining to DG nutrition, and nutrition in various superspecialty cases in critical care settings, where DG is commonly encountered. This paper summarizes the optimum EN practices for managing DG in critically ill patients. The practical solutions to overcome the challenges in this field are presented as practice guidelines at the end of each section. These guidelines are expected to provide guidance for EN management in dysglycemic critically ill patients. These guidelines also outline the model glycemic control task force and its roles in nutrition care as well as an intensive care unit DG nutrition protocol. Mehta Y, Mithal A, Kulkarni A, Reddy BR, Sharma J, Dixit S, et al. Practice Guidelines for Enteral Nutrition Management in Dysglycemic Critically Ill Patients: A Relook for Indian Scenario. Indian J Crit Care Med 2019;23(12):594-603.


Characteristics of facilities with specialized programming for drinking drivers and for other criminal justice involved clients: analysis of a national database.
Offering specialized programming at substance abuse treatment facilities can help diversify clientele and funding sources, potentially enhancing the facilities' ability to survive and/or expand. Past research has shown that facilities only offering specialized programming for driving under the influence/driving while intoxicated offenders (DUI) are predominately private-for-profit owned. As criminal justice populations, both DUI and other criminal justice offenders, comprise a large proportion of those in community-based substance abuse treatment knowing facilities' characteristics would be important for administrators and policymakers to consider when updating programming, training staff or expanding capacity to ensure efficient use of scarce resources. However, while such characteristics are known for DUI programs, they are not known for facilities offering specialized programming for other criminal justice offenders. Analysis of the 2004 US National Survey of Substance Abuse Treatment Facilities. Almost half the facilities (48.2%) offered either DUI or other criminal justice specialized programming. These facilities were divided between those offering DUI specialized programming (17.7%), other criminal justice specialized programming (16.6%) and both types of programming (13.9%). Certain characteristics were independently associated with offering DUI specialized programming (private ownership, rural location, for profit status) or other criminal justice specialized programming (receiving public funds, urban location, region of country). Offering specialized programming for DUI or other criminal justice offenders was common and associated with distinct characteristics. These observed associations may reflect the positioning of the facility to increase visibility, or diversify clientele and possibly funding streams or the decision of policymakers. As the criminal justice populations show no sign of decreasing and resources are scarce, the efficient use of resources demands policymakers recognize the prevalence of these specialized programming, join forces to examine them for efficacy, and explicitly incorporate these characteristics into strategies for workforce training and plans for treatment expansion.


Distal axonopathy in peripheral nerves of PMP22-mutant mice.
A partial duplication of chromosome 17 is associated with Charcot-Marie-Tooth disease type 1A (CMT1A), a demyelinating peripheral neuropathy that causes progressive distal muscle atrophy and sensory impairment. Trisomic expression of peripheral myelin protein 22 (PMP22) whose gene is contained within the duplicated region is considered to be responsible for the disease. By using recombinant gene technology in rodents, we had demonstrated previously that PMP22 is sensitive to gene dosage. Homozygous PMP22 knockout (PMP22(0/0)) mice and transgenic animals carrying additional copies of the PMP22 gene develop distinct peripheral polyneuropathies. We have now performed a detailed morphometrical analysis of the L3 roots, quadriceps and saphenous nerves of these PMP22-mutant mice to study whether the myelin and potential axonal deficits are evenly distributed. The L3 roots and the peripheral nerves were chosen as representatives of the proximal and distal segments of the peripheral nervous system. When the roots were compared with the peripheral nerves, myelin deficiencies appeared more severe at the radicular levels, in particular the ventral roots. Decreased numbers of large calibre axons were a prominent feature in the motor branches of both strains of PMP22-mutant mice, and these axonal deficits were more severe distally. Active axonal damage was only observed in the nerves of PMP22(0/0) mice. Despite the distinct effects on myelination and the Schwann cell phenotype that characterize the neuropathies of PMP22-mutant mice, both strains develop a distally accentuated axonopathy as a common disease mechanism which is likely to be responsible for the neurological deficits.


Novel biogenic aggregation of moss gemmae on a disappearing African glacier.
Tropical regions are not well represented in glacier biology, yet many tropical glaciers are under threat of disappearance due to climate change. Here we report a novel biogenic aggregation at the terminus of a glacier in the Rwenzori Mountains, Uganda. The material was formed by uniseriate protonemal moss gemmae and protonema. Molecular analysis of five genetic markers determined the taxon as Ceratodon purpureus, a cosmopolitan species that is widespread in tropical to polar region. Given optimal growing temperatures of isolate is 20-30 °C, the cold glacier surface might seem unsuitable for this species. However, the cluster of protonema growth reached approximately 10 °C in daytime, suggesting that diurnal increase in temperature may contribute to the moss's ability to inhabit the glacier surface. The aggregation is also a habitat for microorganisms, and the disappearance of this glacier will lead to the loss of this unique ecosystem.


Patient preferences for the integration of mental health counseling and chronic disease care in South Africa.
To describe patient perceptions of the acceptability of integrating mental health counseling within primary care facilities in the Western Cape province of South Africa and their preferences for the way in which this care is delivered. Qualitative interviews with 30 purposively selected patients receiving treatment for HIV or diabetes within primary care facilities who screened positive for depression using the Center for Epidemiological Studies Depression Scale or hazardous alcohol use through the Alcohol Use Disorders Identification Test. Participants articulated high levels of unmet need for mental health services and strong associations between poor mental health and the challenges of living with a chronic disease. Consequently, they considered it acceptable to offer screening and mental health counseling within the context of chronic disease care. They thought counseling would be highly relevant if it helped patients develop adaptive strategies for coping with stress and negative emotions. Irrespective of chronic disease, patients indicated a preference for lay counselors rather than existing clinicians as potential delivery agents, supporting a task-shared approach to mental health counseling delivery in primary care settings. Some expressed concern about the feasibility of using lay counselors already present in facilities to deliver this service, suggesting that additional counselors might be needed. Findings demonstrate a need for mental health counseling within the context of chronic disease care in South Africa. Task-shared approaches, using lay counselors, seem acceptable to patients - provided counselors are selected to ensure they possess the qualities associated with effective counselors. Findings have informed the design of a task-shared mental health program that is responsive to the preferences of patients with chronic diseases.


Global disease burden of conditions requiring emergency surgery.
Surgical disease is inadequately addressed globally, and emergency conditions requiring surgery contribute substantially to the global disease burden. This was a review of studies that contributed to define the population-based health burden of emergency surgical conditions (excluding trauma and obstetrics) and the status of available capacity to address this burden. Further data were retrieved from the Global Burden of Disease Study 2010 and the University of Washington's Institute for Health Metrics and Evaluation online data. In the index year of 2010, there were 896,000 deaths, 20 million years of life lost and 25 million disability-adjusted life-years from 11 emergency general surgical conditions reported individually in the Global Burden of Disease Study. The most common cause of death was complicated peptic ulcer disease, followed by aortic aneurysm, bowel obstruction, biliary disease, mesenteric ischaemia, peripheral vascular disease, abscess and soft tissue infections, and appendicitis. The mortality rate was higher in high-income countries (HICs) than in low- and middle-income countries (LMICs) (24.3 versus 10.6 deaths per 100,000 inhabitants respectively), primarily owing to a higher rate of vascular disease in HICs. However, because of the much larger population, 70 per cent of deaths occurred in LMICs. Deaths from vascular disease rose from 15 to 25 per cent of surgical emergency-related deaths in LMICs (from 1990 to 2010). Surgical capacity to address this burden is suboptimal in LMICs, with fewer than one operating theatre per 100,000 inhabitants in many LMICs, whereas some HICs have more than 14 per 100,000 inhabitants. The global burden of surgical emergencies is described insufficiently. The bare estimates indicate a tremendous health burden. LMICs carry the majority of emergency conditions; in these countries the pattern of surgical disease is changing and capacity to deal with the problem is inadequate. The data presented in this study will be useful for both the surgical and public health communities to plan a more adequate response.


Silencing of host basal defense response-related gene expression increases susceptibility of Nicotiana benthamiana to Clavibacter michiganensis subsp. michiganensis.
Clavibacter michiganensis subsp. michiganensis is an actinomycete, causing bacterial wilt and canker disease of tomato (Solanum lycopersicum). We used virus-induced gene silencing (VIGS) to identify genes playing a role in host basal defense response to C. michiganensis subsp. michiganensis infection using Nicotiana benthamiana as a model plant. A preliminary VIGS screen comprising 160 genes from tomato known to be involved in defense-related signaling identified a set of 14 genes whose suppression led to altered host-pathogen interactions. Expression of each of these genes and three additional targets was then suppressed in larger-scale VIGS experiments and the effect of silencing on development of wilt disease symptoms and bacterial growth during an N. benthamiana-C. michiganensis subsp. michiganensis compatible interaction was determined. Disease susceptibility and in planta bacterial population size were enhanced by silencing genes encoding N. benthamiana homologs of ubiquitin activating enzyme, snakin-2, extensin-like protein, divinyl ether synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase 2, and Pto-like kinase. The identification of genes having a role in the host basal defense-response to C. michiganensis subsp. michiganensis advances our understanding of the plant responses activated by C. michiganensis subsp. michiganensis and raises possibilities for devising novel and effective molecular strategies to control bacterial canker and wilt in tomato.