A portrait of the C4 photosynthetic family on the 50th anniversary of its discovery: species number, evolutionary lineages, and Hall of Fame.
Fifty years ago, the C4 photosynthetic pathway was first characterized. In the subsequent five decades, much has been learned about C4 plants, such that it is now possible to place nearly all C4 species into their respective evolutionary lineages. Sixty-one independent lineages of C4 photosynthesis are identified, with additional, ancillary C4 origins possible in 12 of these principal lineages. The lineages produced ~8100 C4 species (5044 grasses, 1322 sedges, and 1777 eudicots). Using midpoints of stem and crown node dates in their respective phylogenies, the oldest and most speciose C4 lineage is the grass lineage Chloridoideae, estimated to be near 30 million years old. Most C4 lineages are estimated to be younger than 15 million years. Older C4 lineages tend to be more speciose, while those younger than 7 million years have <43 species each. To further highlight C4 photosynthesis for a 50th anniversary snapshot, a Hall of Fame comprised of the 40 most significant C4 species is presented. Over the next 50 years, preservation of the Earth's C4 diversity is a concern, largely because of habitat loss due to elevated CO2 effects, invasive species, and expanded agricultural activities. Ironically, some members of the C4 Hall of Fame are leading threats to the natural C4 flora due to their association with human activities on landscapes where most C4 plants occur.


Magnetic resonance neurography evaluation of chronic extraspinal sciatica after remote proximal hamstring injury: a preliminary retrospective analysis.
Extraspinal sciatica can present unique challenges in clinical diagnosis and management. In this study, the authors evaluated qualitative and quantitative patterns of sciatica-related pathology at the ischial tuberosity on MR neurography (MRN) studies performed for chronic extraspinal sciatica. Lumbosacral MRN studies obtained in 14 patients at the University of California, San Francisco between 2007 and 2011 were retrospectively reviewed. The patients had been referred by neurosurgeons or neurologists for chronic unilateral sciatica (≥ 3 months), and the MRN reports described asymmetrical increased T2 signal within the sciatic nerve at the level of the ischial tuberosity. MRN studies were also performed prospectively in 6 healthy volunteers. Sciatic nerve T2 signal intensity (SI) and cross-sectional area at the ischial tuberosity were calculated and compared between the 2 sides in all 20 subjects. The same measurements were also performed at the sciatic notch as an internal reference. Adjacent musculoskeletal pathology was compared between the 2 sides in all subjects. Seven of the 9 patients for whom detailed histories were available had a specific history of injury or trauma near the proximal hamstring preceding the onset of sciatica. Eight of the 14 patients also demonstrated soft-tissue abnormalities adjacent to the proximal hamstring origin. The remaining 6 had normal muscles, tendons, and marrow in the region of the ischial tuberosity. There was a significant difference in sciatic nerve SI and size between the symptomatic and asymptomatic sides at the level of the ischial tuberosity, with a mean adjusted SI of 1.38 compared with 1.00 (p < 0.001) and a mean cross-sectional nerve area of 0.66 versus 0.54 cm(2) (p = 0.002). The control group demonstrated symmetrical adjusted SI and sciatic nerve size. This study suggests that chronic sciatic neuropathy can be seen at the ischial tuberosity in the setting of prior proximal hamstring tendon injury or adjacent soft-tissue abnormalities. Because hamstring tendon injury as a cause of chronic sciatica remains a diagnosis of exclusion, this distinct category of patients has not been described in the radiographic literature and merits special attention from clinicians and radiologists in the management of extraspinal sciatica. Magnetic resonance neurography is useful for evaluating chronic sciatic neuropathy both qualitatively and quantitatively, particularly in patients for whom electromyography and traditional MRI studies are unrevealing.


Identification of a phosphoprotein that is downregulated in immortalized human fibroblasts.
Many lines of evidence indicate that the immortalization step is critical for the neoplastic transformation of normal human cells. Once normal human cells have been immortalized, they are relatively easily transformed into neoplastic cells. In order to understand these phenomena, patterns of protein phosphorylation in proliferating normal human fibroblast cell strains and their immortalized cell lines were compared by using two-dimensional polyacrylamide gel electrophoresis. It was found that the expression and phosphorylation levels of the human heat shock protein 27 (HSP27) were predominantly downregulated in the immortalized cells compared with those in their normal counterparts. In the normal cells, HSP27 expression and phosphorylation were markedly increased by physiological and nonphysiological stresses, such as serum addition, treatment with a carcinogenic agent like 4-nitroquinoline-1-oxide, and a high osmotic pressure. This may be a normal defense against acute changes of cellular environment and cytotoxic effects. However, these stresses had no effects on the expression and phosphorylation of HSP27 in the immortalized cells. These results suggest that an abnormal regulation of HSP27 expression and phosphorylation may be one of the reasons for easy neoplastic transformation of the immortalized cells by the treatment with carcinogenic agents.


Prevalence and distribution of multilocus sequence types of Staphylococcus aureus isolated from bulk tank milk and cows with mastitis in Pennsylvania.
A total of 163 S. aureus isolates; 113 from mastitic milk (MM) and 50 from bulk tank milk (BTM) (2008, 2013-2015) submitted for bacteriologic analysis at the Penn State Animal Diagnostic Laboratory were examined for their phenotypic and genotypic characteristics. Multi-locus sequence typing (MLST) analysis identified 16 unique sequence types (STs) which belonged to eight clonal complexes (CCs). Majority of the isolates were variants of CC97 (68.7%) and CC151 (25.1%). CC97 comprised of seven STs, of which two were new STs (ST3273, ST3274), while CC151 comprised of three STs of which ST3272 was identified for the first time. Several farms had more than one ST type that were either members of the same clonal complex or unrelated STs. On one farm, six different STs of both categories were seen over the years within the farm. It was observed that ST352 and ST151 were the two main clonal populations in cattle not only in Pennsylvania but also globally. Most isolates were susceptible to all the antibiotics evaluated. 6.7% of isolates showed resistance to vancomycin and penicillin. Two isolates of ST398 showed multidrug resistance (>3 antibiotics) against clindamycin, erythromycin, tetracycline, and penicillin. It was noted that 59 of 163 (36.2%) isolates encoded for enterotoxigenic genes. Enterotoxin genes seg/sei accounted for ~85% of enterotoxin positive isolates. Toxic shock syndrome gene tsst-1 alone was positive in two isolates (ST352, ST 2187). 97.5% of CC151 isolates were enterotoxin seg/sei positive. Most isolates were positive for lukED (95%) and lukAB (96.3%) leukotoxin genes. Bovine specific bi-component leucocidin lukMF' was present in 54% of isolates. A prominent observation of this study was the explicit association of lukMF' with lineages ST151 and ST352. In conclusion, the findings of the study, suggest that small number of S. aureus STs types (ST352, ST2187, ST3028, and ST151) are associated with majority of cases of bovine mastitis in Pennsylvania dairy farms. It was observed that one ST of S. aureus predominated in the herd and this ST can coexist with several other ST types of S. aureus strains. When STs were interpreted along with virulence, leucocidin genes and antimicrobial resistance, ST-variants allowed better interpretation of the S. aureus molecular epidemiologic findings specifically for tracing recurrence or persistence of infections in cow over time, among cows in the herd, and between herds in Pennsylvania.


How far to go? Screening and surveillance in Barrett's esophagus.
There is no dispute that Barrett's esophagus (BE) is associated with an increased risk of esophageal adenocarcinoma. Detecting these cancers early can improve patient survival. But should screening be used to detect BE, or should a surveillance program monitor those already diagnosed with BE for neoplastic changes? Endoscopy and endoscopic biopsy are the only tools available for such screening and surveillance, and the cost effectiveness of either approach must be considered. Two possible solutions are discussed. First, screening could be limited to patients considered at high risk for BE and associated adenocarcinoma. With this approach, more precise risk stratification would be required. The second possible approach is to combine screening for high-risk patients and surveillance for those already diagnosed with BE. Additional outcomes data are needed to determine how often and for what length of time endoscopic surveillance should continue in a patient after several examinations are negative for adenocarcinoma.


[Prediction of maximum oxygen uptake during physical exercise on bicycle ergometer].
Possibilities to predict maximum oxygen uptake (VO2max) during exercising on bicycle ergometer using the Russian Rating Perceived Exertion (RPE) was studied. Results of examination of 13 athletes demonstrate the possibility of predicting individual and group average VO2max on the basis of data from submaximum testing and the empirical formulas from the value of VO2max registered at RPE numbers 13 and 15. These VO2max values can serve as markers for assessing the dynamics of physical performance.


Mitochondrial and nuclear genes suggest that stony corals are monophyletic but most families of stony corals are not (Order Scleractinia, Class Anthozoa, Phylum Cnidaria).
Modern hard corals (Class Hexacorallia; Order Scleractinia) are widely studied because of their fundamental role in reef building and their superb fossil record extending back to the Triassic. Nevertheless, interpretations of their evolutionary relationships have been in flux for over a decade. Recent analyses undermine the legitimacy of traditional suborders, families and genera, and suggest that a non-skeletal sister clade (Order Corallimorpharia) might be imbedded within the stony corals. However, these studies either sampled a relatively limited array of taxa or assembled trees from heterogeneous data sets. Here we provide a more comprehensive analysis of Scleractinia (127 species, 75 genera, 17 families) and various outgroups, based on two mitochondrial genes (cytochrome oxidase I, cytochrome b), with analyses of nuclear genes (ss-tubulin, ribosomal DNA) of a subset of taxa to test unexpected relationships. Eleven of 16 families were found to be polyphyletic. Strikingly, over one third of all families as conventionally defined contain representatives from the highly divergent "robust" and "complex" clades. However, the recent suggestion that corallimorpharians are true corals that have lost their skeletons was not upheld. Relationships were supported not only by mitochondrial and nuclear genes, but also often by morphological characters which had been ignored or never noted previously. The concordance of molecular characters and more carefully examined morphological characters suggests a future of greater taxonomic stability, as well as the potential to trace the evolutionary history of this ecologically important group using fossils.


A fatal case of serotonin syndrome after combined moclobemide-citalopram intoxication.
We present a case involving a fatality due to the combined ingestion of two different types of antidepressants. A 41-year-old Caucasian male, with a history of depression and suicide attempts, was found deceased at home. Multiple containers of medication, the MAO-inhibitor moclobemide (Aurorix), the SSRI citalopram (Cipramil), and the benzodiazepine lormetazepam (Noctamid) as active substance, as well as a bottle of whiskey were present at the scene. The autopsy findings were unremarkable, but systematic toxicological analysis (EMIT, radioimmunoassay, high-performance liquid chromatography-diode-array detection [HPLC-DAD], gas chromatography-nitrogen-phosphorus detection, and gas chromatography-mass spectrometry) revealed the following: ethanol (0.23 g/L blood, 0.67 g/L urine), lormetazepam (1.65 microg/mL urine), cotinine (0.63 microg/mL blood, 5.08 microg/mL urine), caffeine (1.20 microg/mL urine), moclobemide (and metabolites), and citalopram (and metabolite). There upon, we developed a new liquid chromatographic separation with optimized DAD, preceded by an automated solid-phase extraction, for the quantitation of the previously mentioned antidepressive drugs. The results obtained for blood and urine, respectively, were as follows: Ro 12-5637 (moclobemide N'-oxide) not detected and 424 microg/mL; Ro 12-8095 (3-keto-moclobemide) 2.26 microg/mL and 49.7 microg/mL; moclobemide 5.62 microg/mL and 204 microg/mL; desmethylcitalopram 0.42 microg/mL and 1.22 microg/mL; and citalopram 4.47 microg/mL and 19.7 microg/mL. The cause of death was attributed to the synergistic toxicity of moclobemide and citalopram, both antidepressants, which, by intentional or accidental combined ingestion, can produce a potentially lethal hyperserotoninergic state. Based on the history of the case and pharmacology of the drugs involved, the forensic pathologists ruled that the cause of death was multiple drug intoxication, resulting in a fatal "serotonin syndrome," and that the manner of death was suicide.


EEG and ERP biomarkers of Alzheimer's disease: a critical review.
Here we critically review studies that used electroencephalography (EEG) or event-related potential (ERP) indices as a biomarker of Alzheimer's disease. In the first part we overview studies that relied on visual inspection of EEG traces and spectral characteristics of EEG. Second, we survey analysis methods motivated by dynamical systems theory (DST) as well as more recent network connectivity approaches. In the third part we review studies of sleep. Next, we compare the utility of early and late ERP components in dementia research. In the section on mismatch negativity (MMN) studies we summarize their results and limitations and outline the emerging field of computational neurology. In the following we overview the use of EEG in the differential diagnosis of the most common neurocognitive disorders. Finally, we provide a summary of the state of the field and conclude that several promising EEG/ERP indices of synaptic neurotransmission are worth considering as potential biomarkers. Furthermore, we highlight some practical issues and discuss future challenges as well.


Identifying Nonpatient Authors of Patient Portal Secure Messages in Oncology: A Proof-of-Concept Demonstration of Natural Language Processing Methods.
Patient portal secure messages are not always authored by the patient account holder. Understanding who authored the message is particularly important in an oncology setting where symptom reporting is crucial to patient treatment. Natural language processing has the potential to detect messages not authored by the patient automatically. Patient portal secure messages from the Memorial Sloan Kettering Cancer Center were retrieved and manually annotated as a predicted unregistered proxy (ie, not written by the patient) or a presumed patient. After randomly splitting the annotated messages into training and test sets in a 70:30 ratio, a bag-of-words approach was used to extract features and then a Least Absolute Shrinkage and Selection Operator (LASSO) model was trained and used for classification. Portal secure messages (n = 2,000) were randomly selected from unique patient accounts and manually annotated. We excluded 335 messages from the data set as the annotators could not determine if they were written by a patient or proxy. Using the remaining 1,665 messages, a LASSO model was developed that achieved an area under the curve of 0.932 and an area under the precision recall curve of 0.748. The sensitivity and specificity related to classifying true-positive cases (predicted unregistered proxy-authored messages) and true negatives (presumed patient-authored messages) were 0.681 and 0.960, respectively. Our work demonstrates the feasibility of using unstructured, heterogenous patient portal secure messages to determine portal secure message authorship. Identifying patient authorship in real time can improve patient portal account security and can be used to improve the quality of the information extracted from the patient portal, such as patient-reported outcomes.


Use of grape polyphenols against carcinogenesis: putative molecular mechanisms of action using in vitro and in vivo test systems.
Polyphenols are present in foods and beverages and are related to sensorial qualities such as color, bitterness, and astringency, which are relevant in wine, tea, grape juice, and other products. These compounds occur naturally in forms varying from simple phenolic acids to complex polymerized tannins. Thus, it is reasonable to expect that grape-derived products elaborated in the presence of skins and seeds, such as wine and grape juice, are natural sources of flavonoids in the diet. Carcinogenesis is a multistep process that is characterized by genetic, epigenetic, and phenotypic changes. With increasing knowledge of these mechanisms, and the conclusion that most cases of cancer are preventable, efforts have focused on identifying the agents with potential anticancer properties. The use of grape polyphenols against the carcinogenesis process seems to be a suitable alternative for either prevention and/or therapeutic purposes. The aim of this article is to show the molecular data generated from the use of grape polyphenols against carcinogenesis using in vivo and in vitro test systems.


A groupwise association test for rare mutations using a weighted sum statistic.
Resequencing is an emerging tool for identification of rare disease-associated mutations. Rare mutations are difficult to tag with SNP genotyping, as genotyping studies are designed to detect common variants. However, studies have shown that genetic heterogeneity is a probable scenario for common diseases, in which multiple rare mutations together explain a large proportion of the genetic basis for the disease. Thus, we propose a weighted-sum method to jointly analyse a group of mutations in order to test for groupwise association with disease status. For example, such a group of mutations may result from resequencing a gene. We compare the proposed weighted-sum method to alternative methods and show that it is powerful for identifying disease-associated genes, both on simulated and Encode data. Using the weighted-sum method, a resequencing study can identify a disease-associated gene with an overall population attributable risk (PAR) of 2%, even when each individual mutation has much lower PAR, using 1,000 to 7,000 affected and unaffected individuals, depending on the underlying genetic model. This study thus demonstrates that resequencing studies can identify important genetic associations, provided that specialised analysis methods, such as the weighted-sum method, are used.


Experimental measurements of estimator bias and the signal-to-noise ratio for deconvolution from wave-front sensing.
Deconvolution from wave-front sensing (DWFS) has been proposed as a method for achieving high-resolution images of astronomical objects from ground-based telescopes. The technique consists of the simultaneous measurement of a short-exposure focal-plane speckled image, as well as the wave front, by use of a Shack-Hartmann sensor placed at the pupil plane. In early studies it was suspected that some problems would occur in poor seeing conditions; however, it was usually assumed that the technique would work well as long as the wave-front sensor subaperture spacing was less than r(0) (L/r(0) < 1). Atmosphere-induced phase errors in the pupil of a telescope imaging system produce both phase errors and magnitude errors in the effective short-exposure optical transfer function (OTF) of the system. Recently it has been shown that the commonly used estimator for this technique produces biased estimates of the magnitude errors. The significance of this bias problem is that one cannot properly estimate or correct for the frame-to-frame fluctuations in the magnitude of the OTF but can do so only for fluctuations in the phase. An auxiliary estimate must also be used to correct for the mean value of the magnitude error. The inability to compensate for the magnitude fluctuations results in a signal-to-noise ratio (SNR) that is less favorable for the technique than was previously thought. In some situations simpler techniques, such as the Knox-Thompson and bispectrum methods, which require only speckle gram data from the focal plane of the imaging system, can produce better results. We present experimental measurements based on observations of bright stars and the Jovian moon Ganymede that confirm previous theoretical predictions.


Epidemiological features of and changes in incidence of infectious diseases in China in the first decade after the SARS outbreak: an observational trend study.
The model of infectious disease prevention and control changed significantly in China after the outbreak in 2003 of severe acute respiratory syndrome (SARS), but trends and epidemiological features of infectious diseases are rarely studied. In this study, we aimed to assess specific incidence and mortality trends of 45 notifiable infectious diseases from 2004 to 2013 in China and to investigate the overall effectiveness of current prevention and control strategies. Incidence and mortality data for 45 notifiable infectious diseases were extracted from a WChinese public health science data centre from 2004 to 2013, which covers 31 provinces in mainland China. We estimated the annual percentage change in incidence of each infectious disease using joinpoint regression. Between January, 2004, and December, 2013, 54 984 661 cases of 45 infectious diseases were reported (average yearly incidence 417·98 per 100 000). The infectious diseases with the highest yearly incidence were hand, foot, and mouth disease (114·48 per 100 000), hepatitis B (81·57 per 100 000), and tuberculosis (80·33 per 100 000). 132 681 deaths were reported among the 54 984 661 cases (average yearly mortality 1·01 deaths per 100 000; average case fatality 2·4 per 1000). Overall yearly incidence of infectious disease was higher among males than females and was highest among children younger than 10 years. Overall yearly mortality was higher among males than females older than 20 years and highest among individuals older than 80 years. Average yearly incidence rose from 300·54 per 100 000 in 2004 to 483·63 per 100 000 in 2013 (annual percentage change 5·9%); hydatid disease (echinococcosis), hepatitis C, and syphilis showed the fastest growth. The overall increasing trend changed after 2009, and the annual percentage change in incidence of infectious disease in 2009-13 (2·3%) was significantly lower than in 2004-08 (6·2%). Although the overall incidence of infectious diseases was increasing from 2004, the rate levelled off after 2009. Effective prevention and control strategies are needed for diseases with the highest incidence-including hand, foot, and mouth disease, hepatitis B, and tuberculosis-and those with the fastest rates of increase (including hydatid disease, hepatitis C, and syphilis). Chinese Ministry of Science and Technology, National Natural Science Foundation (China).


Alternative water sources and endotoxin.
The primary objective of this preliminary study was to ascertain the concentration of endotoxin in a variety of Australian water types, including recycled water. A total of 42 sampling sites were surveyed, the majority on at least 2 separate occasions (total number of samples analysed was 76). Samples were collected from a variety of locations throughout Australia including: drinking water distribution (12), drinking water reservoir (4), wastewater treatment train (11) and finished recycled water (15). Class A recycled waters, defined microbiologically in Australian regulations as having <10 E. coli per 100 mL, where the treatment train did not include membrane filtration, gave rise to an average measured endotoxin concentration of 2.030 Endotoxin Units (EU) per mL (N=7). For recycled Class A water samples, where membrane filtration was part of the treatment train (N=3) the average endotoxin concentration was 41 EU/mL. Measured endotoxin concentrations in drinking water varied from <4 to 119 EU/mL. Results of this preliminary study indicate that endotoxin concentrations in recycled water may be reduced to levels at least as low as those found in drinking water but for some recycled waters, where membrane filtration is not practiced, higher endotoxin concentrations may persist.


On the nature of emotion regulation.
This paper presents a unitary approach to emotion and emotion regulation, building on the excellent points in the lead article by Cole, Martin, and Dennis (this issue), as well as the fine commentaries that follow it. It begins by stressing how, in the real world, the processes underlying emotion and emotion regulation appear to be largely one and the same, rendering the value of the distinction largely for the benefit of analysis. There is an extensive discussion of how the same processes can generate emotions (i.e., are constitutive of emotion) and account for variability of manifestation of emotion in context (i.e., regulate them). Following an extensive review of many of the principles involved in emotion and emotion regulation, the paper presents implications for developmental study of infants and children, includes several methodological recommendations, and concludes with an analysis of the extent to which contemporary affective neuroscience contributes to the study of emotion and emotion regulation.


[In vivo study of microcirculatory pathways by the contact microscopy technic].
Microcirculatory bed in some organs of a normal brown frog Rana temporaria has been studied at vital observation by means of the luminescent photomicroscope LIUMAM K-1. Elements of the microcirculatory bed have been determined according to the direction of blood stream and the diameter of its separate links. Certain peculiar features of the blood stream have been studied in the dark field in the organs such as: the skin, the tongue, the gastric wall, the intestinal wall, the lungs and others. The range of morpho-functional lability in the microvessels and structural organization of the bed in question depend on the structure of certain organs. A close connection between the microvascular wall and melanocytes has been revealed.


Diaphragm disease after use of nonsteroidal anti-inflammatory agents: first report of diagnosis with capsule endoscopy.
Diaphragm disease of the small intestine is part of the spectrum of diseases associated with injury to the gastrointestinal tract induced by nonsteroidal anti-inflammatory drugs. Standard endoscopy or contrast studies of the small intestine rarely identify these lesions. The diagnosis usually is established at the time of surgery. We report the case of a 72-year-old woman with obscure gastrointestinal bleeding and intermittent obstruction of the small intestine who had had multiple hospitalizations and extensive testing. The patient had been treated with nonsteroidal anti-inflammatory drugs for osteoarthritis. A radiograph of the small intestine with barium contrast revealed no abnormalities, so capsule endoscopy was performed. Capsule endoscopy showed multiple small intestinal strictures beyond which the capsule could not pass. After the patient experienced continued symptoms suggestive of intermittent partial obstruction of the small intestine, computed tomography showed the capsule within a dilated loop of intestine adjacent to a stricture. After 9 days of conservative medical therapy and worsening symptoms, the patient required an exploratory laparotomy. The capsule was located in a 12-cm segment of intestine with 4 diaphragm-like lesions. Pathologic study found submucosal lesions with features identical to those of neuromuscular and vascular hamartoma (eg, mature, reactive tissue elements of smooth muscle, dense fibrous tissue, and nerve tissue bundles with scattered ganglion cells and vessels). No manifestations of Crohn disease were evident. This case represents the first diagnosis with capsule endoscopy of diaphragm disease of the small intestine with pathologic features of neuromuscular and vascular hamartoma.


Ste11p, a high-mobility-group box DNA-binding protein, undergoes pheromone- and nutrient-regulated nuclear-cytoplasmic shuttling.
The high-mobility-group (HMG) box is a conserved DNA-binding domain found in a family of transcription factors that regulate growth and development. One family member, Ste11p, directs sexual differentiation of Schizosaccharomyces pombe by binding specific DNA sequences upstream of genes required for mating and meiosis. Here, we show that Ste11p is a shuttling protein. In growing cells, Ste11p is present in low levels and is pancellular. Mating pheromones and nutrient limitation trigger nuclear accumulation and increased expression of the transcription factor. Several mechanisms likely control Ste11p localization. First, the 14-3-3 protein, Rad24p, binds phosphorylated Ste11p and inhibits its nuclear accumulation. Second, the HMG domain of Ste11p contains a basic cluster nuclear localization signal. Finally, treatment of cells with leptomycin B, an exportin inhibitor, results in the nuclear accumulation of Ste11p. A Ste11p deletion mutation, DeltaC54, mimics the effects of leptomycin B. The C54 region contains no identifiable nuclear export signal but instead is required for biological activity and to stimulate Ste11p target gene expression. These results provide evidence that both nuclear import and export mechanisms operate to regulate cellular localization of an HMG box protein. In addition, they establish a paradigm for the potential role of pheromone/hormone-like polypeptides in cellular localization of this important class of developmental regulators.


Examination of encapsulated phytosterol ester supplementation on lipid indices associated with cardiovascular disease.
As opposed to traditional food based delivery we examined the efficacy of ingesting encapsulated phytosterol esters on indices of lipid health in hypercholesterolemic adults. We performed a randomized, double-blinded, parallel-group, placebo-controlled, clinical intervention examining 54 men and women (20-70 y of age) with a low-density lipoprotein cholesterol (LDL-C) level > or =3.33 mmol/L. Participants were not taking cholesterol-lowering medications. Treatment consisted of ingesting 2.6 g of encapsulated phytosterol esters (n = 25) or a matching placebo (n = 29) for 12 wk. Total cholesterol (TC) levels at baseline (mean +/- SD) were 6.29 +/- 0.7 mmol/L in the phytosterol group and 6.00 +/- 0.7 mmol/L in the placebo group. Baseline LDL-C levels were 4.27 +/- 0.7 mmol/L in the treatment group and 4.00 +/- 0.8 mmol/L in the placebo group. Analysis of variance and Tukey's least significant difference post hoc analyses revealed a significant within-group reduction in TC (-0.23 +/- 0.4 mmol/L, P < 0.05) and LDL-C (-0.22 +/- 0.5 mmol/L, P < 0.05) for the phytosterol treatment group. Mean reductions in TC and LDL-C were greater than placebo (P < 0.05). Percentages of change from baseline for TC were -3.52% (95% confidence interval -6.44 to -0.40) for phytosterol treatment and 2.64% (95% confidence interval 0.30-5.60) for placebo. Those for LDL-C were -5.00% (95% confidence interval -9.92 to -0.08) for phytosterol and 4.89 (95% confidence interval 0.24-9.5) for placebo. No other significant effects were observed. Encapsulated phytosterol ester ingestion appears to positively modulate LDL-C. Given that the reduction in LDL-C was not as extensive as in food-based trials, future investigations should examine potential timing and dose issues relative to encapsulated delivery.


Conducting the train of thought: working memory capacity, goal neglect, and mind wandering in an executive-control task.
On the basis of the executive-attention theory of working memory capacity (WMC; e.g., M. J. Kane, A. R. A. Conway, D. Z. Hambrick, & R. W. Engle, 2007), the authors tested the relations among WMC, mind wandering, and goal neglect in a sustained attention to response task (SART; a go/no-go task). In 3 SART versions, making conceptual versus perceptual processing demands, subjects periodically indicated their thought content when probed following rare no-go targets. SART processing demands did not affect mind-wandering rates, but mind-wandering rates varied with WMC and predicted goal-neglect errors in the task; furthermore, mind-wandering rates partially mediated the WMC-SART relation, indicating that WMC-related differences in goal neglect were due, in part, to variation in the control of conscious thought.


Antipsychotic medication for women with schizophrenia spectrum disorders.
There are significant differences between men and women in the efficacy and tolerability of antipsychotic drugs. Here, we provide a comprehensive overview of what is currently known about the pharmacokinetics and pharmacodynamics of antipsychotics in women with schizophrenia spectrum disorders (SSDs) and translate these insights into considerations for clinical practice. Slower drug absorption, metabolism and excretion in women all lead to higher plasma levels, which increase the risk for side-effects. Moreover, women reach higher dopamine receptor occupancy compared to men at similar serum levels, since oestrogens increase dopamine sensitivity. As current treatment guidelines are based on studies predominantly conducted in men, women are likely to be overmedicated by default. The risk of overmedicating generally increases when sex hormone levels are high (e.g. during ovulation and gestation), whereas higher doses may be required during low-hormonal phases (e.g. during menstruation and menopause). For premenopausal women, with the exceptions of quetiapine and lurasidone, doses of antipsychotics should be lower with largest adjustments required for olanzapine. Clinicians should be wary of side-effects that are particularly harmful in women, such as hyperprolactinaemia which can cause oestrogen deficiency and metabolic symptoms that may cause cardiovascular diseases. Given the protective effects of oestrogens on the course of SSD, oestrogen replacement therapy should be considered for postmenopausal patients, who are more vulnerable to side-effects and yet require higher dosages of most antipsychotics to reach similar efficacy. In conclusion, there is a need for tailored, female-specific prescription guidelines, which take into account adjustments required across different phases of life.


Understanding the antimicrobial activity of selected disinfectants against methicillin-resistant Staphylococcus aureus (MRSA).
Disinfectants and biocidal products have been widely used to combat Methicillin-resistant Staphylococcus aureus (MRSA) infections in homes and healthcare environments. Although disruption of cytoplasmic membrane integrity has been documented as the main bactericidal effect of biocides, little is known about the biochemical alterations induced by these chemical agents. In this study, we used Fourier transform infrared (FT-IR) spectroscopy and chemometric tools as an alternative non-destructive technique to determine the bactericidal effects of commonly used disinfectants against MRSA USA-300. FTIR spectroscopy permits a detailed characterization of bacterial reactivity, allowing an understanding of the fundamental mechanism of action involved in the interaction between bacteria and disinfectants. The disinfectants studied were ethanol 70% (N = 5), isopropanol (N = 5), sodium hypochlorite (N = 5), triclosan (N = 5) and triclocarban (N = 5). Results showed less than 5% colony forming units growth of MRSA treated with triclocarban and no growth in the other groups. Nearly 70,000 mid-infrared spectra from the five treatments and the two control (untreated; N = 4) groups of MRSA (bacteria grown in TSB and incubated at 37°C (Control I) / at ambient temperature (Control II), for 24h) were pre-processed and analyzed using principal component analysis followed by linear discriminant analysis (PCA-LDA). Clustering of strains of MRSA belonging to five treatments and the discrimination between each treatment and two control groups in MRSA (untreated) were investigated. PCA-LDA discriminatory frequencies suggested that ethanol-treated spectra are the most similar to isopropanol-treated spectra biochemically. Also reported here are the biochemical alterations in the structure of proteins, lipid membranes, and phosphate groups of MRSA produced by sodium hypochlorite, triclosan, and triclocarban treatments. These findings provide mechanistic information involved in the interaction between MRSA strains and hygiene products; thereby demonstrating the potential of spectroscopic analysis as an objective, robust, and label-free tool for evaluating the macromolecular changes involved in disinfectant-treated MRSA.


DNA barcoding in pencilfishes (Lebiasinidae: Nannostomus) reveals cryptic diversity across the Brazilian Amazon.
Nannostomus is comprised of 20 species. Popularly known as pencilfishes the vast majority of these species lives in the flooded forests of the Amazon basin and are popular in the ornamental trade. Among the lebiasinids, it is the only genus to have undergone more than one taxonomic revision. Even so, it still possesses poorly defined species. Here, we report the results of an application of DNA barcoding to the identification of pencilfishes and highlight the deeply divergent clades within four nominal species. We surveyed the sequence variation in the mtDNA cytochrome c oxidase subunit I gene among 110 individuals representing 14 nominal species that were collected from several rivers along the Amazon basin. The mean Kimura-2-parameter distances within species and genus were 2% and 19,0%, respectively. The deep lineage divergences detected in N. digrammus, N. trifasciatus, N. unifasciatus and N. eques suggest the existence of hidden diversity in Nannostomus species. For N. digrammus and N. trifasciatus, in particular, the estimated divergences in some lineages were so high that doubt about their conspecific status is raised.


Relative importance of pollen and seed dispersal across a Neotropical mountain landscape for an epiphytic orchid.
Populations of many species are isolated within narrow elevation bands of Neotropical mountain habitat, and how well dispersal maintains genetic connectivity is unknown. We asked whether genetic structure of an epiphytic orchid, Epidendrum firmum, corresponds to gaps between Costa Rican mountain ranges, and how these gaps influence pollen and seed flow. We predicted that significant genetic structure exists among mountain ranges due to different colonization histories and limited gene flow. Furthermore, we predicted that pollen movement contributes more to gene flow than seeds because seeds are released into strong winds perpendicular to the narrow northwest-southeast species distribution, while the likely pollinators are strong fliers. Individuals from 12 populations and three mountain ranges were genotyped with nuclear microsatellites (nDNA) and chloroplast sequences (cpDNA). Genetic diversity was high for both markers, while nDNA genetic structure was low (FSTn = 0.020) and cpDNA structure was moderate (FSTc = 0.443). Significant cpDNA barriers occurred within and among mountain ranges, but nDNA barriers were not significant after accounting for geographic distance. Consistent with these contrasting patterns of genetic structure, pollen contributes substantially more to gene flow among populations than seed (mp /ms = 46). Pollinators mediated extensive gene flow, eroding nDNA colonization footprints, while seed flow was comparatively limited, possibly due to directional prevailing winds across linearly distributed populations. Dispersal traits alone may not accurately inform predictions about gene flow or genetic structure, supporting the need for research into the potentially crucial role of pollinators and landscape context in gene flow among isolated populations.


Transcriptional and epigenetic regulators of human CD8+ T cell function identified through orthogonal CRISPR screens.
Clinical response to adoptive T cell therapies is associated with the transcriptional and epigenetic state of the cell product. Thus, discovery of regulators of T cell gene networks and their corresponding phenotypes has potential to improve T cell therapies. Here we developed pooled, epigenetic CRISPR screening approaches to systematically profile the effects of activating or repressing 120 transcriptional and epigenetic regulators on human CD8+ T cell state. We found that BATF3 overexpression promoted specific features of memory T cells and attenuated gene programs associated with cytotoxicity, regulatory T cell function, and exhaustion. Upon chronic antigen stimulation, BATF3 overexpression countered phenotypic and epigenetic signatures of T cell exhaustion. Moreover, BATF3 enhanced the potency of CAR T cells in both in vitro and in vivo tumor models and programmed a transcriptional profile that correlates with positive clinical response to adoptive T cell therapy. Finally, we performed CRISPR knockout screens that defined cofactors and downstream mediators of the BATF3 gene network.


Dietary pattern in women with polycystic ovary syndrome with and without associated non-alcoholic fatty liver disease: A cross-sectional study.
Women with Polycystic Ovary Syndrome (PCOS) have a higher prevalence of Nonalcoholic Fatty Liver Disease (NAFLD) than the general population. PCOS and NAFLD have common metabolic risk factors, however, the role of diet in NAFLD development is still uncertain in PCOS women. To evaluate and compare the dietary patterns and nutritional intake in patients with PCOS with and without NAFLD. Cross-sectional study that included patients with PCOS diagnosed according to Rotterdam criteria. All participants were submitted to abdominal ultrasound to investigate liver steatosis. Dietary profile was assessed by 24-hour food recall (24hR), and Food Frequency Questionnaire (FFQ). Diet quality was assessed by the Healthy Eating Index (HEI) adapted for the Brazilian population. Physical activity practice was also assessed. 87 participants were included (average age 35.2 ± 5.7 years), among whom, 67 (77%) had NAFLD. The group with PCOS and NAFLD presented higher body mass index (BMI) (34.9 ± 4.5 vs. 30.4 ± 4.9 kg/m2; p = 0.001), Waist Circumference (WC) (103 [97‒113] vs. 95 [87.5‒100] cm; p < 0.001) and were considered physically active less frequently than those without NAFLD (34.3% vs. 60%; p = 0.04). Food intake and dietary patterns assessed by 24hR, FFQ and HEI presented no difference between the groups. PCOS women with coexistent NAFLD had higher BMI, WC and were less physically active than those without NAFLD. Dietary evaluation showed that PCOS women with NAFLD had no significant difference in macro and micronutrients or food group intake and diet quality in comparison to those without NAFLD.


Novel targets in pancreatic cancer research.
The initiation and progression of pancreatic ductal adenocarcinoma (PDA) occurs as a result of molecular alterations that typically result in fluctuations of transcription, protein expression, and ultimately dysregulated signaling pathways. For example, PDA is driven by key activating, gain-of-function mutations in proto-oncogenes (eg, K-Ras) along with loss of function of tumor suppressor genes (eg, p16, SMAD4). With the advent of whole-exome sequencing of PDA genomes, several key genetic alterations have been identified as drivers of PDA. While these findings have led to groundbreaking discoveries in the etiology of PDA, they have failed to provide feasible, targetable therapeutic approaches. Additionally, recent advances in PDA research have uncovered the role of the tumor microenvironment (the non-epithelial tumor cells) in PDA progression by promoting potent, acute changes in gene expression. Herein, this chapter is aimed at discussing the key genetic and non-genetic mechanisms responsible for PDA initiation and progression. Thus based on these mechanisms, we will put forth investigated and novel therapeutic targets in PDA.


Intermittent reloading does not prevent reduction in iron availability and hepcidin upregulation caused by hindlimb unloading.
What is the central question of this study? Could skeletal muscle be involved in microgravity-induced iron misdistribution by modulating expression of hepcidin, the master regulator of iron metabolism? What is the main finding and its importance? We demonstrate, in rats, that hepcidin upregulation is not a transient adaptation associated with early exposure to microgravity and that intermittent reloading does not limit microgravity-induced iron misdistribution despite having a beneficial effect on soleus muscle wasting. In humans, exposure to microgravity during spaceflight causes muscle atrophy, changes in iron storage and a reduction in iron availability. We previously observed that during 7 days of simulated microgravity in rats, hepcidin plays a key role in iron misdistribution, and we suggested that a crosstalk between skeletal muscle and liver could regulate hepcidin synthesis in this context. In the present study in rats, we investigated the medium-term effects of simulated microgravity on iron metabolism. We also tested whether intermittent reloading (IR) to target skeletal muscle atrophy limits iron misdistribution efficiently. For this purpose, Wistar rats underwent 14 days of hindlimb unloading (HU) combined or not combined with daily IR. At the end of this period, the serum iron concentration and transferrin saturation were significantly reduced, whereas hepatic hepcidin mRNA was upregulated. However, the main signalling pathways involved in hepcidin synthesis in the liver (BMP-small mothers against decapentaplegic (SMAD), interleukin-6-STAT3 and ERK1/2) were unaffected. Unlike what was observed after 7 days of HU, the iron concentration in the spleen, liver and skeletal muscle was comparable between control animals and those that underwent HU or HU plus IR for 14 days. Despite its beneficial effect on soleus muscle atrophy and slow-to-fast myosin heavy chain distribution, IR did not significantly prevent a reduction in iron availability and hepcidin upregulation. Altogether, these results highlight that iron availability is durably reduced during longer exposure to simulated microgravity and that the related hepcidin upregulation is not a transient adaptation to these conditions. The results also suggest that skeletal muscle does not necessarily play a key role in the iron misdistribution that occurs during simulated microgravity.


Prolongation of the QT-corrected interval during dobutamine stress echocardiography: a marker for ischemia.
Dobutamine stress echocardiography (DSE) has become a very reliable non-invasive tool in the diagnosis of ischemic heart disease based on the determination of new wall motion abnormalities rather than electrocardiographic changes. In this study, we assessed the usefulness of the corrected QT (QTc) interval and delayed heart rate recovery in predicting ischemia during the infusion of dobutamine. In this retrospective analysis, we analyzed the electrocardiograms of 100 patients who underwent DSE for the diagnosis of coronary artery disease. The QTc interval obtained at peak heart rate during the infusion of dobutamine was compared with the QTc interval at rest. A total of 32 patients had new wall motion abnormalities during DSE, suggestive of ischemia. In these patients, the mean QTc interval at rest was 442.6 ms as compared to 461.0 ms during the peak infusion of dobutamine (P<0.05). In contrast, there was no statistical difference between the mean QTc interval at rest and that noted during DSE in patients without ischemia (439.8 ms and 440.1 ms respectively, P =ls; NS). The QTc interval increased in 40% of patients with ischemia on DSE despite the absence of any accompanying ST-segment depressions. In addition, there was a statistically slower heart rate recovery. Changes in heart rate 2 min into recovery from peak heart rate were 7.3+/-9.5 beats/min in patients with ischemia compared to 12.5+/-11.9 beats/min in those without ischemia (P<0.027). A more significant change was noted 4 min into recovery when compared with the peak heart rate, 14.8+/-10 beats/min in patients with ischemia, compared with 22.2+/-15.7 beats/min in those without ischemia, (P<0.007). These results suggest that the development of new wall motion abnormalities suggestive of ischemia during DSE is associated with prolongation of the QTc interval and delayed heart rate early in the recovery period. These two parameters should be further studied not only as additional markers in the identification of ischemia in patients referred for DSE but also to assess their potential significance during short- and long-term follow-up.


Predictive and prognostic biomarkers with therapeutic targets in breast, colorectal, and non-small cell lung cancers: a systemic review of current development, evidence, and recommendation.
Appropriate evidence-based roles of prognostic and predictive biomarkers of known therapeutic targets in breast, colorectal, and non-small cell lung cancers in adults are reviewed, with summary of evidence for use and recommendation. Current development in biomarker studies is also discussed. Computerized literature searches of PubMed (National Library of Medicine), the Cochrane Collaboration Library, and commonly accepted US and international guidelines (American Society of Clinical Oncology, European Society for Medical Oncology, and National Comprehensive Cancer Network) were performed from 2001 to 2012. Literature published before 2001 was noted for historical interest but not evaluated. Literature review was focused on available systematic reviews and meta-analyses of published predictive (associated with treatment response and/or efficacy) and prognostic (associated with disease outcome) biomarkers of known therapeutic targets in colorectal, breast, and non-small cell lung cancers. In general, significant health outcomes (e.g. predicted response to therapy, overall survival, disease-free survival, quality of life, lesser toxicity, and cost-effectiveness) were used for making recommendations. Four breast cancer biomarkers were evaluated, two of which (2D6 genotyping, Oncotype Dx) were considered emerging with insufficient evidence. Seven colorectal cancer biomarkers were evaluated, five of which (EGFR gene expression, K-ras G13D gene mutation, B-raf V600E gene mutation, dihydropyrimidine dehydrogenase deficiency, and UGT1A1 genotyping) were considered emerging. Seven non-small cell lung cancer biomarkers were evaluated, five of which were emerging (EGFR gene expression, ERCC gene expression, RRM1 gene expression, K-ras gene mutation, and TS gene expression). Of all 18 biomarkers evaluated, the following showed evidence of clinical utility and were recommended for routine use in practice: ER/PR and HER2 for breast cancer; K-ras gene mutation (except G13D gene mutation) for colorectal cancer; mismatch repair deficiency or microsatellite instability for colorectal cancer; and EGFR and EML4-ALK gene mutations for non-small cell lung. Not all recommendations for these biomarkers were uniformly supported by all guidelines.


Reduced reproductive success for a conditioning mutant in experimental populations of Drosophila melanogaster.
Male Drosophila melanogaster that have courted newly-emerged males can modify their subsequent courtship behavior to avoid further courtship with immature males for up to 6 hr (previously reported). Here, it was hypothesized that such an experience-dependent modification would afford a mating advantage to normal males over males that carried a mutation that affects learning and memory. Coisogenic lines were constructed which varied at the dunce gene (dnc+ and dncM14 alleles) in order to test this hypothesis. Whether previously experienced with immature males or not, dnc+ and dncM14 males were indistinguishable in their response and mating efficiency when individually paired with virgin females. However, courtship performance of dnc+ and dncM14 males was different if they were first experienced with immature males and were then individually tested in an artificial population of nine immature males and one virgin female. In this situation, dnc+ males spent much less time in courtship with immature males and achieved copulation in one-third the time required for dncM14 males. As a control, the behavior and mating efficiency of courtship-naive dnc+ and dncM14 males in the artificial population was indistinguishable. In competition for a single virgin female, experienced dncM14 males showed a slight mating advantage over experienced dnc+ males. But when competition by experienced males for a single virgin female took place in the presence of nine immature males, dnc+ males were the successful maters in three-fourths of the trials.


Bayesian semiparametric multiple shrinkage.
High-dimensional and highly correlated data leading to non- or weakly identified effects are commonplace. Maximum likelihood will typically fail in such situations and a variety of shrinkage methods have been proposed. Standard techniques, such as ridge regression or the lasso, shrink estimates toward zero, with some approaches allowing coefficients to be selected out of the model by achieving a value of zero. When substantive information is available, estimates can be shrunk to nonnull values; however, such information may not be available. We propose a Bayesian semiparametric approach that allows shrinkage to multiple locations. Coefficients are given a mixture of heavy-tailed double exponential priors, with location and scale parameters assigned Dirichlet process hyperpriors to allow groups of coefficients to be shrunk toward the same, possibly nonzero, mean. Our approach favors sparse, but flexible, structure by shrinking toward a small number of random locations. The methods are illustrated using a study of genetic polymorphisms and Parkinson's disease.


Heterogeneity in the pharmacological treatment of children with ADHD: cognitive, behavioral, and social functioning differences.
To investigate the extent to which children with ADHD in various medication statuses (i.e., medication naïve, pure stimulant, stimulant plus another medication, nonstimulants) varied on cognitive or academic, behavioral, and social functioning during a psychoeducational assessment battery. Participants for this study consisted of 66 children (20 girls) with a Diagnostic and Statistical Manual of Mental Disorders (4th ed.) (DSM-IV) diagnosis of ADHD confirmed by a comprehensive clinical diagnostic assessment, including the use of a semistructured interview and Conners' Parent Rating Scales. Standardized cognitive and academic measures along with parent report of medication status, behavioral, and social functioning were collected. No differences were found among children in the various medication groups in terms of ADHD symptoms severity, academic performance, processing speed, verbal abilities, or perceptual reasoning skills. However, children in the medication-naïve group performed significantly better than the stimulant-plus-another-medication and nonstimulant groups in terms of overall cognitive abilities, working-memory skills, and social adaptability but had similar scores to children in the pure-stimulant group. Children in the pure-stimulant group also had marginally higher working-memory scores compared to children in the nonstimulant group but not compared to the stimulant-plus-another-medication group. The pure-stimulant group also had significantly lower externalizing and internalizing problems and higher social adaptability compared to the stimulant-plus-another-medication group but not compared to the medication-naïve or nonstimulant groups. Findings showed evidence for distinct cognitive, behavioral, and social profiles among children with ADHD who are proactively not on medication, as well as differences among children with ADHD who are on only one stimulant versus a nonstimulant or stimulant-plus-another-medication regimen.


Relationship between Plantar Pressure and Sensory Disturbance in Patients with Hansen's Disease-Preliminary Research and Review of the Literature.
Orthoses and insoles are among the primary treatments and prevention methods of refractory plantar ulcers in patients with Hansen's disease. While dynamic plantar pressure and tactile sensory disturbance are the critical pathological factors, few studies have investigated whether a relationship exists between these two factors. In this study, dynamic pressure measured using F-scan system and tactile sensory threshold evaluated with monofilament testing were determined for 12 areas of 20 feet in patients with chronic Hansen's disease. The correlation between these two factors was calculated for each foot, for each clinical category of the foot (0-IV) and across all feet. A significant correlation was found between dynamic pressure and tactile sensation in Category II feet (n = 8, p = 0.016, r2 = 0.246, Spearman's rank test). In contrast, no significant correlation was detected for the entire foot or within the subgroups for the remainder of the clinical categories. However, the clinical manifestation of lesion areas showed high variability: (1) pressure concentrated, sensation lost; (2) margin of pressure concentration, sensation lost; (3) pressure concentrated, sensation severely disturbed but not lost; and (4) tip of the toe. These results may indicate that, even though there was a weak relationship between dynamic pressure and tactile sensation, it is important to assess both, in addition to the basics of orthotic treatment in patients with Hansen's disease presenting with refractory plantar ulceration.


Transfer of energy pathway genes in microbial enhanced biological phosphorus removal communities.
Lateral gene transfer (LGT) is an important evolutionary process in microbial evolution. In sewage treatment plants, LGT of antibiotic resistance and xenobiotic degradation-related proteins has been suggested, but the role of LGT outside these processes is unknown. Microbial communities involved in Enhanced Biological Phosphorus Removal (EBPR) have been used to treat wastewater in the last 50 years and may provide insights into adaptation to an engineered environment. We introduce two different types of analysis to identify LGT in EBPR sewage communities, based on identifying assembled sequences with more than one strong taxonomic match, and on unusual phylogenetic patterns. We applied these methods to investigate the role of LGT in six energy-related metabolic pathways. The analyses identified overlapping but non-identical sets of transferred enzymes. All of these were homologous with sequences from known mobile genetic elements, and many were also in close proximity to transposases and integrases in the EBPR data set. The taxonomic method had higher sensitivity than the phylogenetic method, identifying more potential LGTs. Both analyses identified the putative transfer of five enzymes within an Australian community, two in a Danish community, and none in a US-derived culture. Our methods were able to identify sequences with unusual phylogenetic or compositional properties as candidate LGT events. The association of these candidates with known mobile elements supports the hypothesis of transfer. The results of our analysis strongly suggest that LGT has influenced the development of functionally important energy-related pathways in EBPR systems, but transfers may be unique to each community due to different operating conditions or taxonomic composition.


Cost considerations of analgesic therapy: an analysis of the effects of dosing frequency and route of administration.
Using work sampling methodologies, we compared both acquisition and administration costs of equivalent daily doses of controlled-release morphine tablets and morphine sulphate solution. A total cost for each drug therapy was derived by summing: (i) the acquisition cost of the medication, (ii) the cost of drug administration supplies, (iii) the cost of pharmacy time required for packaging and distribution of doses to nursing units, (iv) the cost of nursing time required to administer the medication, (v) the cost of nursing time required to complete the requisite narcotic records. Acquisition costs for controlled-release morphine tablets, morphine sulphate solution and ancillary materials required for dosing were those actually paid by the hospital pharmacy at the time of the study. Costs for nursing and pharmacy time for administration of doses were based on our analysis of the time required to perform each step of the procedure and the median hourly wage for the corresponding position from the British Columbia collective agreement. Assuming a mean total daily morphine dose of 120 mg, administered as one 60 mg controlled-release morphine tablet 12 hourly or 20 mg (4 ml of 5 mg/ml) morphine sulphate solution 4 hourly, morphine solution had a lower acquisition cost than controlled-release tablets ($0.48 vs. $2.32 per day; respectively). However, the saving in acquisition costs was offset by a higher administration cost ($10.20 vs. $2.86 per day). These figures indicate that administration of 120 mg morphine per day to hospitalized patients for 30 days would have a total cost of $155.40 using a regimen to 60 mg controlled-release morphine 12 hourly. In contrast, total cost for a 30 day regimen of 20 mg morphine solution 4 hourly would be $320.40. Thus, an analysis which considers both acquisition and administration costs reveals that the frequency of administration is an important factor in the overall cost of drug therapy and that controlled-release formulations may ultimately be less costly to administer than their immediate release counterparts. A separate analysis of costs associated with the administration of morphine by continuous subcutaneous infusion demonstrated that providing analgesia by this route was significantly more costly than via the oral route.


Cathepsin D precursors in clathrin-coated organelles from human fibroblasts.
Coated vesicles were isolated from metabolically labeled human fibroblasts with the aid of affinity-purified antibodies against human brain clathrin and Staphylococcus aureus cells. The material adsorbed to the S. aureus cells was enriched in clathrin. When the S. aureus cells bearing the immunoadsorbed material were treated with 0.5% saponin, extracts containing the precursor form of cathepsin D were obtained. The extraction of the precursor was promoted in the presence of mannose 6-phosphate. Material adsorbed to S. aureus cells coated with control immunoglobulins was nearly free of clathrin and contained a small amount of the cathepsin D precursor (less than 20% of that adsorbed with anti-clathrin antibodies). The extraction of this cathepsin D precursor was independent of mannose 6-phosphate and was complete after a brief exposure to saponin. The amount of cathepsin D precursor in coated membranes varied between 0.4 and 2.5% of total precursor. Analysis of pulse chase-labeled fibroblasts revealed that cathepsin D was only transiently associated with coated membranes. The mean residence time of cathepsin D precursor in coated membranes was estimated to be 2 min. These observations support the view that coated membranes participate in the transfer of precursor forms of endogenous lysosomal enzymes to lysosomes.


Angiotensin receptor signaling and prostate tumor growth in mice.
The renin-angiotensin system, through its type 1 and type 2 angiotensin receptors (AT1R and AT2R, respectively) may have a role in prostate cancer. The objective of this pilot study was to explore that potential role by determining whether the AT1R blocker, losartan, would reduce the growth of LAPC-4 prostate cancer xenografts in nude mice. We also evaluated the tumor growth effects of using angiotensin II to activate both AT1R and AT2R simultaneously. Our data showed that losartan decreased tumor volumes by 56% versus control. This decrease reached statistical significance at day 54 (p = 0.0014). By day 54, Ki67 was also reduced in the losartan group, though not significantly so (p = 0.077). Losartan had no significant effect on AT1R or AT2R expression. Despite significant increases in both AT1R and AT2R at day 29 (p = 0.043 and 0.038, respectively), the administration of angiotensin II did not result in any significant differences in tumor volumes or ki67 at any time point. These data suggest that selective activation and induction of AT2R coupled with blockade of AT1R may slow prostate cancer growth. Future larger studies are needed to confirm these results.


Competitive Association of Antibiotics with a Clay Mineral and Organoclay Derivatives as a Control of Their Lifetimes in the Environment.
A Na-smectite clay mineral (Na-Mt) was exchanged with two concentrations of benzyldimethyltetradecyl ammonium chloride cationic surfactant up to one time the cation exchange capacity. Nonionic organoclay was prepared with polyoxyethylene (20) oleyl ether (Brij-O20) nonionic surfactant at one concentration. The resulting organoclays displayed lateral layer organization of the surfactants within their interlayer space.. The adsorption properties of these organoclays and the starting raw clay mineral were evaluated for three extensively used antibiotic pharmaceutical products: the amoxicillin (AMX), the sulfamethoxazole (SMX), and the trimethoprim (TRI), recognized as recalcitrant compounds to conventional water treatments and to display a complex behavior for different pH and temperature experimental conditions. Besides showing short half-life time with possible degradation by UV radiation, these antibiotics associated with mineral phases cause serious environmental issues of which the toxic effect can be exacerbated in the presence of other chemical compounds. From the set of data obtained by complementary techniques: UV and Fourier transform infrared spectroscopy, high-performance liquid chromatography coupled with mass spectrometry, and X-ray diffraction, it appears that the nonionic organoclay shows its versatility for the adsorption of individual molecules as well as a pool of antibiotics. The mixing of the three antibiotics showing different electric charged species (cations, anions, and zwitterions) mimics the natural context drives to a deep modification of the adsorption behavior onto the different materials that can act as possible carrier mineral phases in aquatic environment. These competition effects can be measured through the significant decrease of the K F Freundlich constants for AMX in the presence of other molecules (or electrolytes), whereas TRI and SMX, by their possible association, create a synergistic effect that favors their adsorption on the whole layered materials.


Maize flour fortification in Africa: markets, feasibility, coverage, and costs.
The economic feasibility of maize flour and maize meal fortification in Kenya, Uganda, and Zambia is assessed using information about the maize milling industry, households' purchases and consumption levels of maize flour, and the incremental cost and estimated price impacts of fortification. Premix costs comprise the overwhelming share of incremental fortification costs and vary by 50% in Kenya and by more than 100% across the three countries. The estimated incremental cost of maize flour fortification per metric ton varies from $3.19 in Zambia to $4.41 in Uganda. Assuming all incremental costs are passed onto the consumer, fortification in Zambia would result in at most a 0.9% increase in the price of maize flour, and would increase annual outlays of the average maize flour-consuming household by 0.2%. The increases for Kenyans and Ugandans would be even less. Although the coverage of maize flour fortification is not likely to be as high as some advocates have predicted, fortification is economically feasible, and would reduce deficiencies of multiple micronutrients, which are significant public health problems in each of these countries.


Evaluation of an established pericardium patch for delivery of mesenchymal stem cells to cardiac tissue.
The present study has evaluated a commercial pericardial material for its capacity to assist as a natural extracellular matrix (ECM) patch for the delivery and retention of mesenchymal stem cells for cardiac repair. The repair of cardiac tissue with cells delivered by an appropriate bioscaffold is expected to offer a superior, long-lasting treatment strategy. The present material, CardioCel®, is based on acellular pericardium that has been stabilized by treatments, including a low concentration of glutaraldehyde, that eliminate calcification after implantation. In the present study, we have assessed this material using human bone marrow mesenchymal stem cells at various cell densities under standard, static cell culture conditions. The initial seeding densities were monitored to evaluate the extent of cell attachment and cell viability, with subsequent cell proliferation assessed up to 4 weeks using an MTS assay. Cell morphology, infiltration, and spreading were tracked using scanning electron microscopy and phalloidin staining. The efficacy of long-term cell survival was further assessed by examining the extent and type of new tissue formation on seeded scaffolds at 70 days; both type I and type III collagens were present in fibrillar structures on these scaffolds indicating that the seeded stem cells had the capacity to differentiate into collagen-producing cells necessary to repair damaged ECM. These data show that the CardioCel® scaffold is an appropriate substrate for the stem cells and has the potential to both retain seeded stem cells and to act as a template for cell propagation and new tissue formation.


Chloroquine and hydroxychloroquine in the management of COVID-19: Much kerfuffle but little evidence.
Chloroquine and hydroxychloroquine are drugs that have shown in vitro activity on the replication of certain coronaviruses. In the context of the SARS-Cov-2 epidemic, the virus responsible for the novel coronavirus disease (COVID-19), these two drugs have been proposed as possible treatments. The results of the first clinical studies evaluating the effect of hydroxychloroquine do not support any efficacy of this drug in patients with COVID-19, due to major methodological weaknesses. Yet, these preliminary studies have aroused considerable media interest, raising fears of massive and uncontrolled use. In the absence of evidence of clinical benefits, the main risk is of exposing patients unnecessarily to the well-known adverse effects of hydroxychloroquine, with a possibly increased risk in the specific setting of COVID-19. In addition, widespread use outside of any recommendation risks compromising the completion of good quality clinical trials. The chloroquine hype, fueled by low-quality studies and media announcements, has yielded to the implementation of more than 150 studies worldwide. This represents a waste of resources and a loss of opportunity for other drugs to be properly evaluated. In the context of emergency, rigorous trials are more than ever needed in order to have, as soon as possible, reliable data on drugs that are possibly effective against the disease. Meanwhile, serious adverse drug reactions have been reported in patients with COVID-19 receiving hydroxychloroquine, justifying to limit its prescription, and to perform suitable cardiac and therapeutic drug monitoring.


JURaSSiC: accuracy of clinician vs risk score prediction of ischemic stroke outcomes.
We compared the accuracy of clinicians and a risk score (iScore) to predict observed outcomes following an acute ischemic stroke. The JURaSSiC (Clinician JUdgment vs Risk Score to predict Stroke outComes) study assigned 111 clinicians with expertise in acute stroke care to predict the probability of outcomes of 5 ischemic stroke case scenarios. Cases (n = 1,415) were selected as being representative of the 10 most common clinical presentations from a pool of more than 12,000 stroke patients admitted to 12 stroke centers. The primary outcome was prediction of death or disability (modified Rankin Scale [mRS] ≥3) at discharge within the 95% confidence interval (CI) of observed outcomes. Secondary outcomes included 30-day mortality and death or institutionalization at discharge. Clinicians made 1,661 predictions with overall accuracy of 16.9% for death or disability at discharge, 46.9% for 30-day mortality, and 33.1% for death or institutionalization at discharge. In contrast, 90% of the iScore-based estimates were within the 95% CI of observed outcomes. Nearly half (n = 53 of 111; 48%) of participants were unable to accurately predict the probability of the primary outcome in any of the 5 rated cases. Less than 1% (n = 1) provided accurate predictions in 4 of the 5 cases and none accurately predicted all 5 case outcomes. In multivariable analyses, the presence of patient characteristics associated with poor outcomes (mRS ≥3 or death) in previous studies (older age, high NIH Stroke Scale score, and nonlacunar subtype) were associated with more accurate clinician predictions of death at 30 days (odds ratio [OR] 2.40, 95% CI 1.57-3.67) and with a trend for more accurate predictions of death or disability at discharge (OR 1.85, 95% CI 0.99-3.46). Clinicians with expertise in stroke performed poorly compared to a validated tool in predicting the outcomes of patients with an acute ischemic stroke. Use of the risk stroke outcome tool may be superior for decision-making following an acute ischemic stroke.


Clinical Profiles of Children with Disruptive Behaviors Based on the Severity of Their Conduct Problems, Callous-Unemotional Traits and Emotional Difficulties.
This study identified clinical profiles of referred children based on the severity of callous-unemotional (CU) traits, emotional difficulties, and conduct problems. Parents of 166 children (132 males) aged 6-12 years referred to a hospital clinic because of disruptive behavior completed measures to assess these key indicators, and person-centered analysis was used to identify profiles. Four distinct profiles were identified that include: (1) Children low in severity on the three domains, (2) Children high in severity on the three domains, (3) Children high in severity in conduct problems and CU traits with minimal emotional difficulties, and (4) Children high in severity in conduct problems and emotional difficulties with minimal CU traits. Profiles differed in degree of aggression and behavioral impairment. Findings show that clinic-referred children with disruptive behaviors can be grouped based on these important indicators into profiles that have important implications for assessment and treatment selection.


Spatial pulse modulation.
Pulse modulation of electronic time signals is a well-proven concept, particularly in the context of data communications and computers. We suggest that pulse modulation of spatial optical signals might be another fruitful concept. The optical components and optical methods for performing the necessary operations for spatial pulse modulation are available. We demonstrate this experimentally by encoding a two-dimensional analog signal (an image) into a one-dimensional pulse signal. This might be of interest in connection with picture transmission over electronic channels, with cinematography, and with holography.


The absorption of phosphate ions from the ovine reticulorumen.
The reticulorumen is now recognised to be an important site of net absorption of phosphate ions from ruminal fluid containing phosphate concentrations appropriate to those found in normal farming practice. These rates of absorption were measured in vivo from solutions placed in the washed reticulorumen, isolated in situ, in conscious, trained sheep. Reducing the ruminal sodium concentration led to reduced absorption of phosphate, suggestive that phosphate and sodium fluxes across the apical wall of the ruminal epithelial cell are linked, as they are in the kidney. Increased absorption of short chain fatty acids led to enhanced absorption of phosphate ions. Conversely, inhibition of carbonic anhydrase activity, by the addition of 1 mM acetazolamide to the ruminal fluid, led to a reduction in phosphate absorption. An increase in the acidity of the ruminal fluid also increased the absorption of phosphate, as did an increase in the ruminal Ca(2+) concentration over the range 1-4 mmol per litre. It is suggested that these effects can be accounted for by a Na(+)/H(+) antiporter coupled with a phosphate/proton symporter in the apical membrane of the ruminal epithelial cell.


The Emerging Threat of (Micro)Thrombosis in COVID-19 and Its Therapeutic Implications.
The recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the ensuing global pandemic has presented a health emergency of unprecedented magnitude. Recent clinical data has highlighted that coronavirus disease 2019 (COVID-19) is associated with a significant risk of thrombotic complications ranging from microvascular thrombosis, venous thromboembolic disease, and stroke. Importantly, thrombotic complications are markers of severe COVID-19 and are associated with multiorgan failure and increased mortality. The evidence to date supports the concept that the thrombotic manifestations of severe COVID-19 are due to the ability of SARS-CoV-2 to invade endothelial cells via ACE-2 (angiotensin-converting enzyme 2), which is expressed on the endothelial cell surface. However, in patients with COVID-19 the subsequent endothelial inflammation, complement activation, thrombin generation, platelet, and leukocyte recruitment, and the initiation of innate and adaptive immune responses culminate in immunothrombosis, ultimately causing (micro)thrombotic complications, such as deep vein thrombosis, pulmonary embolism, and stroke. Accordingly, the activation of coagulation (eg, as measured with plasma D-dimer) and thrombocytopenia have emerged as prognostic markers in COVID-19. Given thrombotic complications are central determinants of the high mortality rate in COVID-19, strategies to prevent thrombosis are of critical importance. Several antithrombotic drugs have been proposed as potential therapies to prevent COVID-19-associated thrombosis, including heparin, FXII inhibitors, fibrinolytic drugs, nafamostat, and dipyridamole, many of which also possess pleiotropic anti-inflammatory or antiviral effects. The growing awareness and mechanistic understanding of the prothrombotic state of COVID-19 patients are driving efforts to more stringent diagnostic screening for thrombotic complications and to the early institution of antithrombotic drugs, for both the prevention and therapy of thrombotic complications. The shifting paradigm of diagnostic and treatment strategies holds significant promise to reduce the burden of thrombotic complications and ultimately improve the prognosis for patients with COVID-19.


Fine particulate matter measurements in Swiss restaurants, cafés and bars: what is the effect of spatial separation between smoking and non-smoking areas?
We performed 124 measurements of particulate matter (PM(2.5)) in 95 hospitality venues such as restaurants, bars, cafés, and a disco, which had differing smoking regulations. We evaluated the impact of spatial separation between smoking and non-smoking areas on mean PM(2.5) concentration, taking relevant characteristics of the venue, such as the type of ventilation or the presence of additional PM(2.5) sources, into account. We differentiated five smoking environments: (i) completely smoke-free location, (ii) non-smoking room spatially separated from a smoking room, (iii) non-smoking area with a smoking area located in the same room, (iv) smoking area with a non-smoking area located in the same room, and (v) smoking location which could be either a room where smoking was allowed that was spatially separated from non-smoking room or a hospitality venue without smoking restriction. In these five groups, the geometric mean PM(2.5) levels were (i) 20.4, (ii) 43.9, (iii) 71.9, (iv) 110.4, and (v) 110.3 microg/m(3), respectively. This study showed that even if non-smoking and smoking areas were spatially separated into two rooms, geometric mean PM(2.5) levels in non-smoking rooms were considerably higher than in completely smoke-free hospitality venues. PM(2.5) levels are considerably increased in the non-smoking area if smoking is allowed anywhere in the same location. Even locating the smoking area in another room resulted in a more than doubling of the PM(2.5) levels in the non-smoking room compared with venues where smoking was not allowed at all. In practice, spatial separation of rooms where smoking is allowed does not prevent exposure to environmental tobacco smoke in nearby non-smoking areas.


Differential Responders to a Mixed Meal Tolerance Test Associated with Type 2 Diabetes Risk Factors and Gut Microbiota-Data from the MEDGI-Carb Randomized Controlled Trial.
The global prevalence of type 2 diabetes mellitus (T2DM) has surged in recent decades, and the identification of differential glycemic responders can aid tailored treatment for the prevention of prediabetes and T2DM. A mixed meal tolerance test (MMTT) based on regular foods offers the potential to uncover differential responders in dynamical postprandial events. We aimed to fit a simple mathematical model on dynamic postprandial glucose data from repeated MMTTs among participants with elevated T2DM risk to identify response clusters and investigate their association with T2DM risk factors and gut microbiota. Data were used from a 12-week multi-center dietary intervention trial involving high-risk T2DM adults, comparing high- versus low-glycemic index foods within a Mediterranean diet context (MEDGICarb). Model-based analysis of MMTTs from 155 participants (81 females and 74 males) revealed two distinct plasma glucose response clusters that were associated with baseline gut microbiota. Cluster A, inversely associated with HbA1c and waist circumference and directly with insulin sensitivity, exhibited a contrasting profile to cluster B. Findings imply that a standardized breakfast MMTT using regular foods could effectively distinguish non-diabetic individuals at varying risk levels for T2DM using a simple mechanistic model.


Measles in Africa: a deadly disease. Some personal comments.
Measles dominate childhood illnesses in tropical Africa with a variety and severity of complications virtually unknown in the West. There is no apparent continental variation in measles virus but environmental factors vary widely. The frequent association of severe measles with malnutrition and recent studies indicating impaired humoral, cellular, and mucocutaneous immunologic function in the malnourished may explain this perplexing disease. Prevention of early malnutrition combined with lasting and economic early immunization with heat-stable vaccine, currently unavailable, are needed to halt this frequent costly, and often fatal African scourge.


[The arteriovenous difference in osmotic and hemostatic indices as a criterion for the early diagnosis of pulmonary complications in severe combined trauma].
The arteriovenous difference in the values of plasma and its main ingredients' osmolality and hemostasis were examined in patients with grave combined injuries complicated with the adult respiratory distress syndrome (RDS) in the posttraumatic period. The findings showed a changed trend of arteriovenous difference in the osmotic homeostasis parameters in comparison with those during on uneventful course of combined injury: a-v Dosm. in stage IV RDS is (-)22.9 +/- 13.5 mosm/kg H2O versus 3.36 +/- 3.51 mosm/kg H2O in an uncomplicated course (p < 0.001). The arteriovenous difference for plasma COD was 8.1 +/- 3.0 mm Hg and (-)0.5 +/- 0.7 mm Hg in an uncomplicated course (p < 0.001). These changes permit a conclusion about the impaired osmolality regulating function of the lungs in RDS. Analysis of the arteriovenous difference of hemostasis once again confirms the presence of disorders in the hemostasis regulating function of the lungs: the arteriovenous difference of the structural and chronometric values of thromboelastogram is inverted, which indicates an increase of the coagulation potential of arterial blood: the a-v is equal to difference of blood clotting time 48 h after the injury and is (-)0.1 +/- 0.04 min, in contrast to the uneventful course: 2.5 +/- 0.27 min (p < 0.01): the a-v equal to the coagulation index difference was 2.26 +/- 0.49 arb. U, in contrast to (-)2.0 +/- 0.13 arb. U in an uncomplicated course of heating. Disorders in the other than respiratory functions of the lungs develop 11 h earlier than clear-cut x-ray signs of RDS appear, which gives us grounds to consider these changes as the earliest diagnostic criterion of RDS.


Whole-brain blood flow and oxygen metabolism in the rat during nitrous oxide anesthesia.
The Kety-Schmidt washout technique has been modified to measure whole-brain blood flow and metabolism in the rat. During nitrous oxide anesthesia, 14 rats exhaled (133)Xe, and continuous and simultaneous arterial and cerebral venous samples were drawn from a femoral artery and the transverse sinus of the brain. Extracerebral contamination of the venous sample was minimal, and equilibration of (133)Xe in brain tissue and blood was obtained after 10-24 min of inhalation. Cerebral blood flow was calculated from the total activity of the mechanically integrated arterial and venous samples according to the principle of Scheinberg and Stead. At a mean Paco2 of 40 mmHg, CBF averaged 98 +/- 6 (SEM) ml/100 g-min and CMRO2 averaged 5.4 +/- 0.7 (SEM) ml/100 g-min. CBF changed 2.4% with each millimeter Hg change of Paco2 while CMRO2 changed only insignificantly. The values obtained for CBF are higher than reported for man and large laboratory animals bur reflect the proportionately greater amount of gray matter in the rat brain.


Role of 5-HT on increased permeability of blood-brain barrier under heat stress.
Exposure of young rats (9-10 wks) to chronic summer heat (36 degrees C) or acute heat (38 degrees C 4hr) increased the BBB permeability to Evans blue albumin complex (MW 68,000) and 131I-sodium (MW 154) in different brain regions which correlated well with the increased level of 5-HT in plasma and brain. This increased permeability of BBB and the increased 5-HT level were prevented by pretreatment with p-CPA, indomethacin and diazepam. Cyproheptadine and vinblastine pretreatment however, prevented only the increased permeability of BBB, the plasma and brain 5-HT level continued to remain high. These results indicate a probable role of 5-HT as one of the factors leading to the increased permeability of BBB in young rats following heat stress.


Photochemical Behavior of Microbial Extracellular Polymeric Substances in the Aquatic Environment.
Microbially derived extracellular polymeric substances (EPSs) occupy a large portion of dissolved organic matter (DOM) in surface waters, but the understanding of the photochemical behaviors of EPS is still very limited. In this study, the photochemical characteristics of EPS from different microbial sources (Shewanella oneidensis, Escherichia coli, and sewage sludge flocs) were investigated in terms of the production of reactive species (RS), such as triplet intermediates (3EPS*), hydroxyl radicals (•OH), and singlet oxygen (1O2). The steady-state concentrations of •OH, 3EPS*, and 1O2 varied in the ranges of 2.55-8.73 × 10-17, 3.01-4.56 × 10-15, and 2.08-2.66 × 10-13 M, respectively, which were within the range reported for DOM from other sources. The steady-state concentrations of RS varied among different EPS isolates due to the diversity of their composition. A strong photochemical degradation of the protein-like components in EPS isolates was identified by excitation emission matrix fluorescence with parallel factor analysis, but relatively, humic-like components remained stable. Fourier-transform ion cyclotron resonance mass spectrometry further revealed that the aliphatic portion of EPS was resistant to irradiation, while other portions with lower H/C ratios and higher O/C ratios were more susceptible to photolysis, leading to the phototransformation of EPS to higher saturation and lower aromaticity. With the phototransformation of EPS, the RS derived from EPS could effectively promote the degradation of antibiotic tetracycline. The findings of this study provide new insights into the photoinduced self-evolution of EPS and the interrelated photochemical fate of contaminants in the aquatic environment.


Effect of intraoperative intravenous crystalloid infusion on postoperative nausea and vomiting after gynaecological laparoscopy: comparison of 30 and 10 ml kg(-1).
I.V. fluid administration has been shown to reduce postoperative nausea and vomiting (PONV). The optimum dose is unknown. We tested the hypothesis that administration of i.v. crystalloid of 30 ml kg(-1) would reduce the incidence of PONV compared with 10 ml kg(-1) of the same fluid. A total of 141 ASA I female patients undergoing elective gynaecological laparoscopy were randomized, in double-blind fashion, to receive either 10 ml kg(-1) (n=71; CSL-10 group) or 30 ml kg(-1) (n=70; CSL-30 group) of i.v. compound sodium lactate (CSL). In the first 48 h after anaesthesia, the incidence of vomiting was lower in the CSL-30 group than in the CSL-10 group (8.6% vs 25.7%, P=0.01). Anti-emetic use was less in the CSL-30 group at 0.5 h (2.9% vs 14.3%, P=0.04). The incidence of severe nausea was significantly reduced in the treatment group at awakening (2.9% vs 15.7%, P=0.02), 2 h (0.0% vs 8.6%, P=0.04) and cumulatively (5.7% vs 27.1%, P=0.001). The numbers needed to treat to prevent vomiting, severe nausea and antiemetic use in the first 48 h were 6, 5 and 6, respectively. I.V. administration of CSL 30 ml kg(-1) to healthy women undergoing day-case gynaecological laparoscopy reduced the incidence of vomiting, nausea and anti-emetic use when compared with CSL 10 ml kg(-1).


Mitogenomes of Giant-Skipper Butterflies reveal an ancient split between deep and shallow root feeders.
Background: Giant-Skipper butterflies from the genus Megathymus are North American endemics. These large and thick-bodied Skippers resemble moths and are unique in their life cycles. Grub-like at the later stages of development, caterpillars of these species feed and live inside yucca roots. Adults do not feed and are mostly local, not straying far from the patches of yucca plants. Methods: Pieces of muscle were dissected from the thorax of specimens and genomic DNA was extracted (also from the abdomen of a specimen collected nearly 60 years ago). Paired-end libraries were prepared and sequenced for 150bp from both ends. The mitogenomes were assembled from the reads followed by a manual gap-closing procedure and a phylogenetic tree was constructed using a maximum likelihood method from an alignment of the mitogenomes. Results: We determined mitogenome sequences of nominal subspecies of all five known species of Megathymus and Agathymus mariae to confidently root the phylogenetic tree. Pairwise sequence identity indicates the high similarity, ranging from 88-96% among coding regions for 13 proteins, 22 tRNAs and 2 rRNA, with a gene order typical for mitogenomes of Lepidoptera. Phylogenetic analysis confirms that Giant-Skippers (Megathymini) originate within the subfamily Hesperiinae and do not warrant a subfamily rank. Genus Megathymus is monophyletic and splits into two species groups. M. streckeri and M. cofaqui caterpillars feed deep in the main root system of yucca plants and deposit frass underground. M. ursus, M. beulahae and M. yuccae feed in the yucca caudex and roots near the ground, and deposit frass outside through a "tent" (a silk tube projecting from the center of yucca plant). M. yuccae and M. beulahae are sister species consistently with morphological similarities between them. Conclusions: We constructed the first DNA-based phylogeny of the genus Megathymus from their mitogenomes. The phylogeny agrees with morphological considerations.


Bloodborne markers in humans during multiday exposure to ozone.
Intermittent exposure of the human lung to ambient levels of ozone (O3) was assayed in systemic fluids by using serum alpha-tocopherol (ST) as a gauge of oxidative stress and the blastogenic activity of peripheral blood monocytes as an index of immune function. Healthy men (n = 10) were evaluated over 3 consecutive days (130 min/day) of chamber exposure to O3 and filtered air (FA); subjects alternated between rest and light treadmill exercise during exposures. For O3, the level was varied at 20-min intervals, i.e., 250, 350, 450, 450, 350, and 250 parts/billion, and concluded with 10 min at 250 parts/billion. ST was quantitated by high-performance liquid chromatography techniques, and T-lymphocyte blastogenesis was measured in cell cultures of peripheral blood monocytes by comparing [3H]thymidine incorporation in mitogen-stimulated (concanavalin A) and nonstimulated cells. After the third day of O3 at 20 h postexposure, ST levels were reduced significantly compared with the FA control subjects (down 14%; -0.96 mumol/l). Mitogen-activated T lymphocytes exhibited a 61% increase in blastogenic activity after 3 days of O3 exposure, significant compared with the proliferative activity of activated T lymphocytes collected after FA or before O3. Acute airway function was impaired by O3, e.g., on day 1, the forced vital capacity and forced expiratory volume in 1 s were decreased 8% (-0.92 liter) and 14% (-0.86 l/s), respectively, from preexposure values, and full recovery was delayed beyond 24 h. Effects of O3 exposure on cellular and biochemical markers increased in magnitude after each exposure and did not parallel the apparent adaptability of bronchial sensitivity to O3.


Order of amino acid limitation in meat and bone meal.
Three experiments were conducted to determine the order of amino acid (AA) limitation in meat and bone meal (MBM) using AA addition and deletion assays. In two addition assays, various individual and combined additions of eight AA were made to semipurified basal diets containing 16% protein solely from a MBM. The two MBM had previously been determined to vary greatly in protein quality. In the deletion assay, a semipurified basal diet containing 13.5% CP provided solely by MBM was supplemented with L-Thr, L-Val, DL-Met, L-Leu, L-Ile, L-Phe, L-Tyr, L-Lys-HCl, L-His, and L-Trp to fulfill the Illinois Ideal Protein or AA pattern on a digestible basis. Each AA was then deleted individually from the basal diet, and the effect of its deletion on growth performance was assessed. In both addition assays, supplementation of the MBM basal diet with Trp and Cys together yielded a large increase in growth performance, whereas supplementation with these AA individually had no effect. These results indicated that Trp and sulfur AA (with a primary need for Cys) were equally first limiting in the MBM. The order of limitation for the other AA was unclear. The results of the deletion assay showed that deletion of Trp, Thr, Phe + Tyr, Ile, Met, Lys, Val, or His significantly depressed growth performance. The results of the combined addition and deletion experiments indicated that the order of AA limitation in MBM was 1) Trp and sulfur AA, 2) Thr, 3) Ile and Phe + Tyr, 4) Met, 5) Lys, and 6) Val and His. The deletion assay using diets formulated on an ideal protein basis was more effective than the addition assay for determining the order of AA limitation in MBM.


Children with developmental and behavioural concerns in Singapore.
Childhood developmental and behavioural disorders (CDABD) have been increasingly recognised in recent years. This study evaluated the profiles and outcomes of children referred for developmental and behavioural concerns to a tertiary child developmental centre in Singapore. This is the first such regional database. Baseline information, obtained through a questionnaire, together with history at first consultation, provided information for referral, demographic and presentation profiles. Clinical formulations were then made. Definitive developmental and medical diagnoses, as well as outcomes based on clinical assessment and standardised testing, were recorded at one year post first consultation. Out of 1,304 referrals between January 1, 2003 and December 1, 2004, 45% were 2-4 years old and 74% were boys. The waiting time from referral to first consultation exceeded four months in 52% of children. Following clinical evaluation, 7% were found to be developmentally appropriate. The single most common presenting concern was speech and language (S&L) delay (29%). The most common clinical developmental diagnosis was autism spectrum disorder (ASD) (30%), followed by isolated S&L disorder, global developmental delay (GDD) and cognitive impairment (CI). Recommendations included S&L therapy (57%), occupational therapy (50%) and psychological/behavioural services (40%). At one year, ASD remained the most common definitive developmental diagnosis (31%), followed by S&L disorder, CI and GDD. Most were children with high-prevalence, low-moderate severity disorders who could potentially achieve fair-good prognosis with early intervention. Better appreciation of the profile and outcome of children with CDABD in Singapore could enable better resource planning for diagnosis and intervention.