Multi-fidelity prediction of molecular optical peaks with deep learning.
Optical properties are central to molecular design for many applications, including solar cells and biomedical imaging. A variety of ab initio and statistical methods have been developed for their prediction, each with a trade-off between accuracy, generality, and cost. Existing theoretical methods such as time-dependent density functional theory (TD-DFT) are generalizable across chemical space because of their robust physics-based foundations but still exhibit random and systematic errors with respect to experiment despite their high computational cost. Statistical methods can achieve high accuracy at a lower cost, but data sparsity and unoptimized molecule and solvent representations often limit their ability to generalize. Here, we utilize directed message passing neural networks (D-MPNNs) to represent both dye molecules and solvents for predictions of molecular absorption peaks in solution. Additionally, we demonstrate a multi-fidelity approach based on an auxiliary model trained on over 28 000 TD-DFT calculations that further improves accuracy and generalizability, as shown through rigorous splitting strategies. Combining several openly-available experimental datasets, we benchmark these methods against a state-of-the-art regression tree algorithm and compare the D-MPNN solvent representation to several alternatives. Finally, we explore the interpretability of the learned representations using dimensionality reduction and evaluate the use of ensemble variance as an estimator of the epistemic uncertainty in our predictions of molecular peak absorption in solution. The prediction methods proposed herein can be integrated with active learning, generative modeling, and experimental workflows to enable the more rapid design of molecules with targeted optical properties.


Environmental and ventilation assessment in Child Day Care Centers in Porto: the ENVIRH Project.
Children attending day care centers (CDCC) have been reported to be more prone to infectious diseases when compared with those cared for at home, and are exposed to conditions that may increase the risk of allergies and asthma. Several studies revealed that consequences of poor ventilation conditions include high levels of carbon dioxide (CO2) and many other indoor pollutants commonly detected in schools. Nine child day care centers were selected randomly to participate in this study. Fifty-two classrooms were assessed for chemical, biological, physical, and allergen parameters in spring and winter seasons in these nine CDCC located in Porto, Portugal. Outdoor measurements were also conducted for comparison. Our results indicated that (i) particulate matter (PM10) median levels were above the national reference levels, both by classroom type and by season; (ii) TVOC kindergarten peak values may raise some concern; (iii) CO2 was present at high median and maximum levels during spring and winter assessment in both nurseries and kindergartens classrooms; (iv) total bacteria concentrations were 57- and 52-fold higher in the nursery and kindergarten than outdoors, respectively, for the spring season; (v) winter and spring median predicted mean vote (PMV) indices were between "neutral" (0) and "slightly cool" (≤ -1) in the thermal sensation scale for comfort situations (-2 to 2) for both types of classrooms; (vi) there were significant differences for both PMV and predicted percentage of dissatisfied (PPD) indices by season; and (vii) CO2, total bacteria, and gram-negative bacteria were associated with low airflow rates. These data will help to evaluate the effectiveness of current building operation practices in child day care centers regarding indoor air quality and respiratory health.


Transcutaneous vagal nerve stimulatio (t-VNS): An adjunctive treatment option for refractory epilepsy.
The aim of this trial was to investigate the efficacy and safety of transcutaneous vagal nerve stimulation (t-VNS) in the palliative treatment of drug resistant epileptic patients ineligible for surgery. Twenty adult patients received four hours of t-VNS per day for six months (T1), followed by a two-month washout period (T2). The frequency and type of seizures recorded at T1 and T2 were compared with those occurring in the three months preceding study entry (T0). Responders (>30% reduction in the total number of seizures) subsequently received two hours of t-VNS per day for further six months (T3). All patients underwent electroencephalography (EEG) and completed the Quality of Life in Epilepsy questionnaire at baseline and T1. At T1 six patients were considered responders. In these patients, at T3 the average reduction in seizure frequency was 60% compared to T0 (p = 0.043), and 51% compared to T2 (p = 0.043). Responders had more often seizures with falls (5 of 6; 83.3%) compared with non-responders (3 of 14; 21.4%) (p = 0,010) and t-VNS reduced their frequency by a percentage ranging from 47.5 to 100%. There was no change in responders' EEG findings after stimulation. At the end of the trial, three responders continued t-VNS, one implanted VNS. t-VNS had no or minimal side effects and significantly reduced seizures in about one third of the enrolled patients. Further studies should be planned to assess whether t-VNS is a suitable tool to predict the efficacy of implanted VNS.


Fluidization and wall slip of soft glassy materials by controlled surface roughness.
We present a comprehensive study of concentrated emulsions flowing in microfluidic channels, one wall of which is patterned with micron-size equally spaced grooves oriented perpendicularly to the flow direction. We find a scaling law describing the roughness-induced fluidization as a function of the density of the grooves, thus fluidization can be predicted and quantitatively regulated. This suggests common scenarios for droplet trapping and release, potentially applicable for other jammed systems as well. Numerical simulations confirm these views and provide a direct link between fluidization and the spatial distribution of plastic rearrangements.


The potential of inhibitors of endocannabinoid metabolism as anxiolytic and antidepressive drugs--A practical view.
The endocannabinoid system, comprising cannabinoid CB1 and CB2 receptors, their endogenous ligands anandamide and 2-arachidonoylglyerol, and their synthetic and metabolic enzymes, are involved in many biological processes in the body, ranging from appetite to bone turnover. Compounds inhibiting the breakdown of anandamide and 2-arachidonoylglycerol increase brain levels of these lipids and thus modulate endocannabinoid signalling. In the present review, the preclinical evidence that these enzymes are good targets for development of novel therapies for anxiety and depression are discussed from a practical, rather than mechanistic, point of view. It is concluded that the preclinical data are promising, albeit tempered by problems of tolerance as well as effects upon learning and memory for irreversible monoacylglycerol lipase inhibitors, and limited by a focus upon male rodents alone. Clinical data so far has been restricted to safety studies with inhibitors of anandamide hydrolysis and a hitherto unpublished study on such a compound in elderly patients with major depressive disorders, but under the dose regimes used, they are well tolerated and show no signs of "cannabis-like" behaviours.


Measuring on-line compliance in ventilated infants using hot wire anemometry.
We investigated the validity of using tidal volume (VT) as measured by the neonatal volume monitor (NVM) to derive respiratory compliance. The NVM is a noninvasive device that measures VT by hot wire anemometry. The quotient of VT and the inflation pressure amplitude from the mechanical ventilator provided a measure of respiratory compliance. This was validated against the single breath occlusion technique in 15 infants (birth weight 0.9 to 4.4 kg). To ensure fully passive expiration, only paralyzed or sedated patients were studied. Only 12 of the 15 infants were analyzed because of limitations in the single breath technique. In three infants the flow-volume curves obtained were alinear, indicating inhomogeneity. In the 12 infants with acceptable single breath data, agreement between the two methods was excellent. Using the expired volume, r2 was .99. We conclude that the NVM can be used to obtain valid estimates of respiratory compliance on-line in intubated infants.


Physical tactics of female partners against male batterer program participants.
Descriptive and predictive analyses were conducted using a multisite database of batterer program participants to assess the nature and extent of their female partner's violence, and implications for batterer program outcome (N = 563). Approximately 40% of the women reported ever using "severe" conflict tactics on the Conflict Tactics Scale (CTS; 17% used severe tactics in the arrest incident). Approximately 20% of the women reported using any type of physical tactic during a 15-month follow-up, and nearly all of these women were with men who physically attacked them during that period. These women were also highly likely to report acting out of fear or self-defense, and having sought a variety of services to deal with the men's violence. Their male partners, furthermore, showed evidence of antisocial tendencies and alcohol problems. Overall, the findings suggest women's "violent resistance" rather than "mutuality and symmetry." Batterer programs appear more appropriate in this regard than couples counseling.


Prevention of hemolysis-induced organ damage by nutritional activation of the vagal anti-inflammatory reflex*.
Acute hemolysis is associated with organ damage, inflammation, and impaired vascular function. Stimulation of the cholecystokinin-1 receptor-dependent vagal anti-inflammatory reflex with lipid-rich enteral nutrition was demonstrated to prevent tissue damage and attenuate inflammation. This study investigates the effects of nutritional activation of the vagal anti-inflammatory reflex on organ integrity, systemic inflammation, and microcirculation during hemolysis. Prospective randomized controlled study. University research unit. Male Sprague-Dawley rats. Intravascular hemolysis was simulated by infusion of prelysed erythrocytes. Animals were fasted or received lipid-rich enteral nutrition. Pegylated (PEG)-CCK9A, A70104 (a cholecystokinin-1 receptor antagonist), and chlorisondamine (a nicotinic acetylcholine receptor antagonist) were applied to investigate involvement of the vagal reflex. Nutritional intervention reduced hemolysis-related renal tubular cell damage, hepatocyte damage, ileal leakage of horseradish peroxidase, and bacterial translocation compared with food deprivation (all p < 0.05). Also circulating interleukin (IL)-6 levels were decreased by enteral nutrition (p < 0.05). Blockage of the cholecystokinin-1 receptor or the nicotinic acetylcholine receptor reversed the protective nutritional effects compared with vehicle (p < 0.05), whereas PEG-CCK9 mimicked the impact of enteral feeding in fasted animals (p < 0.05). Furthermore, nutritional intervention increased renal, hepatic, and intestinal blood flow compared with fasting (all p < 0.05), as evaluated using fluorescent microspheres. Nutritional activation of the vagal anti-inflammatory reflex preserves tissue integrity and attenuates systemic inflammation in a rodent model of acute hemolysis. In addition, lipid-rich nutrition improves renal, hepatic, and intestinal microcirculation. These findings implicate stimulation of the autonomic nervous system by nutritional means as a potential therapy to prevent complications of acute hemolysis. (Crit Care Med 2013; 41:e361-e367).


Genome size, secondary simplification, and the evolution of the brain in salamanders.
Compared to other vertebrates, even including lampreys and hagfishes in some respects, salamanders exhibit a relatively simple organization of brain and sense organs which is illustrated here using the visual system as an example. The greatest simplicity is found in the bolitoglossine salamanders, yet all bolitoglossines possess highly projectile tongues and rely on vision for survival; furthermore, some species are agile and acrobatic. The unusual features of the visual system of salamanders include small numbers of large neurons, a low degree of morphological differentiation among neurons, a small proportion of myelinated axons in the optic nerve, and an optic tectum consisting essentially of a periventricular cellular layer and a superficial fiber layer. Similar features are found throughout the central nervous system of salamanders and in the lateral line, auditory and olfactory systems as well. Phylogenetic analysis shows that the most parsimonious interpretation of these data is that the simple organization of the brain and sense organs of salamanders was derived secondarily from a more complex ancestral state. We hypothesize that increased genome size has led to simplification of the nervous system in salamanders. Increased genome size appears to have had profound effects on neural development in salamanders, leading to paedomorphosis, the retention of juvenile or even embryonic characteristics into adulthood. In particular, large genome size is associated with large cell size and reduced rates of cell proliferation, migration and differentiation. Secondary simplification has constrained the function of the salamanders' visual system, primarily by increasing cell size and decreasing cell numbers. However, it also has provided an opportunity for the evolution of compensating mechanisms, which have helped to restore or even enhance visual function. Most apparent among the compensatory mechanisms of bolitoglossine salamanders is the presence of well developed ipsilateral retinotectal projections, which apparently enhance depth perception. It is difficult to explain the unusual history of the nervous system in salamanders solely in terms of natural selection and adaptation. Increasing genome size through selfish replication appears to have played a major role in the evolution of salamander brains by imposing functional constraints as well as creating opportunities for overcoming them.


Widespread, focal copy number variations (CNV) and whole chromosome aneuploidies in Trypanosoma cruzi strains revealed by array comparative genomic hybridization.
Trypanosoma cruzi is a protozoan parasite and the etiologic agent of Chagas disease, an important public health problem in Latin America. T. cruzi is diploid, almost exclusively asexual, and displays an extraordinarily diverse population structure both genetically and phenotypically. Yet, to date the genotypic diversity of T. cruzi and its relationship, if any, to biological diversity have not been studied at the whole genome level. In this study, we used whole genome oligonucleotide tiling arrays to compare gene content in biologically disparate T. cruzi strains by comparative genomic hybridization (CGH). We observed that T. cruzi strains display widespread and focal copy number variations (CNV) and a substantially greater level of diversity than can be adequately defined by the current genetic typing methods. As expected, CNV were particularly frequent in gene family-rich regions containing mucins and trans-sialidases but were also evident in core genes. Gene groups that showed little variation in copy numbers among the strains tested included those encoding protein kinases and ribosomal proteins, suggesting these loci were less permissive to CNV. Moreover, frequent variation in chromosome copy numbers were observed, and chromosome-specific CNV signatures were shared by genetically divergent T. cruzi strains. The large number of CNV, over 4,000, reported here uphold at a whole genome level the long held paradigm of extraordinary genome plasticity among T. cruzi strains. Moreover, the fact that these heritable markers do not parse T. cruzi strains along the same lines as traditional typing methods is strongly suggestive of genetic exchange playing a major role in T. cruzi population structure and biology.


An assessment of the association between smoking status, pain intensity, and functional interference in patients with chronic pain.
This article examines the association between smoking and pain intensity and functional interference in a heterogeneous group of patients evaluated at a tertiary outpatient pain clinic. Current smoking is associated with less favorable clinical presentations. This study was conducted to determine if the smoking status of patients seen in an outpatient pain clinic is associated with differences in pain intensity and interference. Surveys were mailed to 500 consecutive new patients evaluated at an outpatient pain clinic. Measures included the Brief Pain Inventory (BPI) and the Fagerström Test for Nicotine Dependence (FTND). Univariate analyses compared BPI scores between smokers and non-smokers. Mean BPI scores were compared between smoking status via analysis of covariance (adjusted for demographic variables which differed significantly by smoking status). A p value < or = 0.05 was accepted as significant. Survey completion rate was 46%, and 14.7% were current smokers. Smokers were younger, and more likely to be male and unemployed. Smokers had higher scores on all the pain intensity BPI scales (p < 0.01), and higher scores (indicating greater functional impairment) on the general activity (p = 0.007), mood (p = 0.003), normal work (p = 0.02), relationships (p = 0.04), sleep (p < 0.001), and life enjoyment (p = 0.03) BPI functional impairment scales. Severe nicotine dependence was associated with greater pain now, (p = 0.05), and greater functional interference on mood (p = 0.005), normal work (p = 0.02) and life enjoyment (p = 0.04) BPI scales. In patients who completed evaluation in an outpatient pain clinic, current cigarette smokers reported significantly greater pain intensity and pain interference with functioning. Symptoms were more pronounced in smokers with more severe nicotine dependence.


Benefit of Cisplatin With Definitive Radiotherapy in Older Women With Cervical Cancer.
Cisplatin with definitive radiotherapy (RT) is considered the standard of care for cervical cancer; however, older women are frequently undertreated and have worse outcomes compared with younger patients. Because women aged ≥65 years have been disproportionately underrepresented in clinical trials, uncertainties exist regarding how much they benefit from the addition of cisplatin to RT. Women aged ≥65 years with nonmetastatic cervical cancer treated with definitive external-beam RT and brachytherapy were identified in the SEER-Medicare database. Death attributable to cervical cancer (cancer-specific mortality [CSM]) was evaluated against competing risks of death using Gray's test. Propensity score analysis and the Fine-Gray multivariable regression model were used to adjust for baseline differences, including comorbidity. The total cohort comprised 826 patients, of whom 531 (64%) received cisplatin, 233 (28%) were FIGO stage I, 374 (45%) were stage II, and 219 (27%) were stage III-IVA. Older age and chronic kidney disease significantly predicted omission of cisplatin. Virtually all cisplatin dosing was weekly, with a median of 5 cycles. Death from cervical cancer was significantly lower with cisplatin than without (5-year CSM, 31% vs 39%; P=.02; adjusted hazard ratio, 0.72; P=.02), which persisted in propensity score analysis. Receiving ≥5 cycles was required for benefit, as no difference in CSM was seen in patients receiving 1 to 4 cycles versus no cisplatin. Subgroup analyses revealed that the benefit of cisplatin persisted in women aged ≥75 years and those with early-stage disease. Incidence of cytopenia, nausea/vomiting, and hypovolemia increased in patients treated with cisplatin. Administration of cisplatin with definitive RT in women aged ≥65 years was associated with a significant benefit in the incidence of death attributable to cervical cancer, despite competing risks for mortality in an older population. Receiving at least 5 cycles of weekly cisplatin was required for benefit.


Mechanisms linking exposure to preeclampsia in utero and the risk for cardiovascular disease.
Preeclampsia (PE) is now recognised as a cardiovascular risk factor for women. Emerging evidence suggests that children exposed to PE in utero may also be at increased risk of cardiovascular disease (CVD) in later life. Individuals exposed to PE in utero have higher systolic and diastolic blood pressure and higher body mass index (BMI) compared to those not exposed to PE in utero. The aim of this review is to discuss the potential mechanisms driving the relationship between PE and offspring CVD. Exposure to an adverse intrauterine environment as a consequence of the pathophysiological changes that occur during a pregnancy complicated by PE is proposed as one mechanism that programs the fetus for future CVD risk. Consistent with this hypothesis, animal models of PE where progeny have been studied demonstrate causality for programming of offspring cardiovascular health by the preeclamptic environment. Shared alleles between mother and offspring, and shared lifestyle factors between mother and offspring provide alternate pathways explaining associations between PE and offspring CVD risk. In addition, adverse lifestyle habits can also act as second hits for those programmed for increased CVD risk. PE and CVD are both multifactorial diseases and, hence, identifying the relative contribution of PE to offspring risk for CVD is a very complex task. However, considering the emerging strong association between PE and CVD, those exposed to PE in utero may benefit from targeted primary CVD preventive strategies.


Kit signaling is involved in melanocyte stem cell fate decisions in zebrafish embryos.
Adult stem cells are crucial for growth, homeostasis and repair of adult animals. The melanocyte stem cell (MSC) and melanocyte regeneration is an attractive model for studying regulation of adult stem cells. The process of melanocyte regeneration can be divided into establishment of the MSC, recruitment of the MSC to produce committed daughter cells, and the proliferation, differentiation and survival of these daughter cells. Reduction of Kit signaling results in dose-dependent reduction of melanocytes during larval regeneration. Here, we use clonal analysis techniques to develop assays to distinguish roles for these processes during zebrafish larval melanocyte regeneration. We use these clonal assays to investigate which processes are affected by the reduction in Kit signaling. We show that the regeneration defect in kita mutants is not due to defects in MSC recruitment or in the proliferation, differentiation or survival of the daughter cells, but is instead due to a defect in stem cell establishment. Our analysis suggests that the kit MSC establishment defect results from inappropriate differentiation of the MSC lineage.


Autochthonous dermal leishmaniasis in Texas.
Over a 2-year period two cases of dermal leishmaniasis of special interest were recognized in Texas. The first occurred in 1972 in a 74-year-old woman who had residences in Dilworth and Gahzales, Gonzales County, and the other was in a 56-year-old man from Kenedy, Karnes County, in 1974. Both cases were biopsy- and culture-positive, and the second patient exhibited anti-Leishmania antibodies by indirect immunofluorescent antibody and direct agglutination tests. Epidemiologic investigation revealed no association between the two cases and suggested that both patients had acquired their infections locally in southern Texas. Moreover, serologic evidence of Leishmania infection was uncovered in a neighbor of the second case and in 3 dogs living nearby. Potential sylvatic reservoirs and arthropod vectors of the disease are resident in the area. Epidemiologic data suggest that dermal leishmaniasis is endemic in south-central Texas.


Formation of ectopic neurepithelium in chick blastoderms: age-related capacities for induction and self-differentiation following transplantation of quail Hensen's nodes.
Hensen's node, regarded as the avian and mammalian homologue of Spemann's neural inducer (i.e., the amphibian dorsal blastoporal lip), has been transplanted in many previous studies to the germinal crescent of avian blastoderms to examine ectopic neural induction. All these studies have suffered from one or more major shortcomings, the most significant of which has been the lack of a reliable cell marker to determine the contributions of graft cells to ectopic embryos. In the absence of such marker, induced (i.e., derived from the host) and self-differentiated (i.e., derived from the graft) neurepithelium cannot be distinguished from one another with certainty. We have transplanted quail Hensen's nodes to chick host blastoderms and have subsequently used the quail nucleolar heterochromatin marker to identify graft cells unequivocally. We systematically varied both donor and host ages (i.e., stages 3-8 and 3-5, respectively) to examine the effects of age on ectopic neural induction and self-differentiation. Our results demonstrate that the age of the donor is more critical than that of the host over the stages examined. With advancing donor age, the frequency of host induction decreases, while the frequency of graft self-differentiation increases. Previous studies not using cell markers have concluded that the craniocaudal level of the induced neuraxis is determined by the age of the donor, that is, young donors induce cranial neuraxial levels, whereas old donors induce caudal levels. By contrast, we found that with grafts from older donors, neurepithelium was more commonly self-differentiated rather than induced and that progressively more caudal levels of the neuraxis self-differentiated with advancing donor age. Induction of caudal neuraxial levels never occurred in the absence of induced cranial levels. The frequency of neural induction was inversely correlated with the age of the donor and directly correlated with the quantity of graft endodermal cells contributed to the ectopic embryo, supporting a previous assertion that in avian embryos, the earliest and principal source of neural inducer lies within the endoderm rather than mesoderm. From our results, we propose that the role of neural induction is to produce neurepithelium of unspecified regional character, and that the formation of regional character depends on subsequent morphogenetic events.


Detection of resistance mutations to antivirals oseltamivir and zanamivir in avian influenza A viruses isolated from wild birds.
The neuraminidase (NA) inhibitors oseltamivir and zanamivir are the first-line of defense against potentially fatal variants of influenza A pandemic strains. However, if resistant virus strains start to arise easily or at a high frequency, a new anti-influenza strategy will be necessary. This study aimed to investigate if and to what extent NA inhibitor-resistant mutants exist in the wild population of influenza A viruses that inhabit wild birds. NA sequences of all NA subtypes available from 5490 avian, 379 swine and 122 environmental isolates were extracted from NCBI databases. In addition, a dataset containing 230 virus isolates from mallard collected at Ottenby Bird Observatory (Öland, Sweden) was analyzed. Isolated NA RNA fragments from Ottenby were transformed to cDNA by RT-PCR, which was followed by sequencing. The analysis of genotypic profiles for NAs from both data sets in regard to antiviral resistance mutations was performed using bioinformatics tools. All 6221 sequences were scanned for oseltamivir- (I117V, E119V, D198N, I222V, H274Y, R292K, N294S and I314V) and zanamivir-related mutations (V116A, R118K, E119G/A/D, Q136K, D151E, R152K, R224K, E276D, R292K and R371K). Of the sequences from the avian NCBI dataset, 132 (2.4%) carried at least one, or in two cases even two and three, NA inhibitor resistance mutations. Swine and environmental isolates from the same data set had 18 (4.75%) and one (0.82%) mutant, respectively, with at least one mutation. The Ottenby sequences carried at least one mutation in 15 cases (6.52%). Therefore, resistant strains were more frequently found in Ottenby samples than in NCBI data sets. However, it is still uncertain if these mutations are the result of natural variations in the viruses or if they are induced by the selective pressure of xenobiotics (e.g., oseltamivir, zanamivir).


Shifting Paradigms Revisited: Biotechnology and the Pharmaceutical Sciences.
In 2003, Crommelin et al. published an article titled: "Shifting paradigms: biopharmaceuticals versus low molecular weight drugs" (https://doi.org/10.1016/S0378-5173(03)00376-4). In the present commentary, 16 years later, we discuss pharmaceutically relevant aspects of the evolution of biologics since then. First, we discuss the increasing repertoire of biologics, in particular, the rapidly growing monoclonal antibody family and the advent of advanced therapy medicinal products. Next, we discuss trends in formulation and characterization as well as summarize our current insights into immunogenicity of biologics. We spend a separate section on new product(ion) paradigms for biologics, such as cell-free production systems, production of advanced therapy medicinal products, and downscaled production approaches. Furthermore, we share our views on issues related to reaching the patient, including routes and techniques of administration, alternative development models for affordable biologics, biosimilars, and handling of biologics. In the concluding section, we outline outstanding issues and make some suggestions for resolving those.


Self-medication of mental health problems: new evidence from a national survey.
To evaluate the association between past 30-day use of alcohol, marijuana, and other illicit drugs and past year unmet need for and use of mental health care. A subsample of 18,849 respondents from the 2001 National Household Survey on Drug Abuse and the 2002 National Survey on Drug Use and Health. Subjects were between the ages of 18 and 65 years and had least one past year mental disorder symptom and no past year substance dependency. Logistic regressions of past 30-day substance use on past 12-month unmet need for mental health care and past 12-month use of mental health services controlling for clinical and sociodemographic characteristics. Predicted probabilities and corresponding standard errors are reported. Use of illicit drugs other than marijuana increased with unmet need for mental health care (4.4 versus 3.2 percent, p=.046) but was not reduced with mental health-care use. Heavy alcohol use was not associated with increased unmet need for mental health care, but was higher among individuals with no mental health care use (4.4 percent versus 2.7 percent, p<.001). By contrast, marijuana use did not appear associated with either unmet need or mental health care use. Substance use varies with past year unmet need for mental health care and mental health care use in ways consistent with the self-medication hypothesis. Results suggest that timely screening and treatment of mental health problems may prevent the development of substance-use disorders among those with mental disorders. Further research should identify subgroups of individuals for whom timely and appropriate mental health treatment would prevent the development of substance-use disorders.


Achromatic design in the illumination system for a mini projector with LED light source.
We provide a novel achromatic design in the illumination system for a mini projector with an LED light source. The lateral color aberration at the corners of the DMD active area can be reduced to 0.48 μm by our compact ATIR prism. The total prism size is 4091 mm(3). The spot sizes at the corner and the edge are also controlled under 198 μm. Moreover, we use a light pipe for color uniformity at the center of the DMD active area. Under the two components for color uniformity, the Δu' v' is well controlled under 0.016 by a 20 mm length of light pipe. The illuminance uniformity at the active area of the DMD chip is higher than 92.8%. The optical efficiency of the light pipe is 94.1%.


Computed tomography angiography of situs inversus, portosystemic shunt and multiple vena cava anomalies in a dog.
A 5-year-old Shih Tzu was presented with intermittent vomiting and anorexia. Microhepatica and reversed position of the abdominal organs were observed on radiography. Ultrasonographically, portosystemic shunt (PSS) was tentatively diagnosed. Computed tomography (CT) revealed that the distended portal vein drained into the left hepatic vein. The caudal vena cava (CdVC) split postrenally and converged at the renal level. Cranial to this, the azygos continuation of the CdVC was confirmed. In the thorax, a persistent left cranial vena cava (CrVC) was found along with right CrVC. This is the first report of a dog with persistent left CrVC and multiple abdominal malformations. CT angiography was useful in evaluating the characteristics of each vascular anomaly and determining the required surgical correction in this complex case.


Changes in Posture Following a Single Session of Long-Duration Water Immersion.
Transitioning between different sensory environments is known to affect sensorimotor function and postural control. Water immersion presents a novel environmental stimulus common to many professional and recreational pursuits, but is not well-studied with regard to its sensorimotor effects upon transitioning back to land. The authors investigated the effects of long-duration water immersion on terrestrial postural control outcomes in veteran divers. Eleven healthy men completed a 6-hour thermoneutral pool dive (4.57 m) breathing diver air. Center of pressure was observed before and 15 minutes after the dive under 4 conditions: (1) eyes open/stable surface (Open-Stable); (2) eyes open/foam surface (Open-Foam); (3) eyes closed/stable surface (Closed-Stable); and (4) eyes closed/foam surface (Closed-Foam). Postdive decreases in postural sway were observed in all testing conditions except for Open-Stable. The specific pattern of center of pressure changes in the postdive window is consistent with (1) a stiffening/overregulation of the ankle strategy during Open-Foam, Closed-Stable, and Closed-Foam or (2) acute upweighting of vestibular input along with downweighting of somatosensory, proprioceptive, and visual inputs. Thus, our findings suggest that postimmersion decreases in postural sway may have been driven by changes in weighting of sensory inputs and associated changes in balance strategy following adaptation to the aquatic environment.


Mineralocorticoid receptor-mediated vascular insulin resistance: an early contributor to diabetes-related vascular disease?
Two-thirds of adults in the U.S. are overweight or obese, and another 26 million have type 2 diabetes (T2D). Patients with diabetes and/or the metabolic syndrome have a significantly increased risk of heart attack and stroke compared with people with normal insulin sensitivity. Decreased insulin sensitivity in cardiovascular tissues as well as in traditional targets of insulin metabolic signaling, such as skeletal muscle, is an underlying abnormality in obesity, hypertension, and T2D. In the vasculature, insulin signaling plays a critical role in normal vascular function via endothelial cell nitric oxide production and modulation of Ca(2+) handling and sensitivity in vascular smooth muscle cells. Available evidence suggests that impaired vascular insulin sensitivity may be an early, perhaps principal, defect of vascular function and contributor to the pathogenesis of vascular disease in persons with obesity, hypertension, and T2D. In the overweight and obese individual, as well as in persons with hypertension, systemic and vascular insulin resistance often occur in concert with elevations in plasma aldosterone. Indeed, basic and clinical studies have demonstrated that elevated plasma aldosterone levels predict the development of insulin resistance and that aldosterone directly interferes with insulin signaling in vascular tissues. Furthermore, elevated plasma aldosterone levels are associated with increased heart attack and stroke risk. Conversely, renin-angiotensin-aldosterone system and mineralocorticoid receptor (MR) antagonism reduces cardiovascular risk in these patient populations. Recent and accumulating evidence in this area has implicated excessive Ser phosphorylation and proteosomal degradation of the docking protein, insulin receptor substrate, and enhanced signaling through hybrid insulin/IGF-1 receptor as important mechanisms underlying aldosterone-mediated interruption of downstream vascular insulin signaling. Prevention or restoration of these changes via blockade of aldosterone action in the vascular wall with MR antagonists (i.e., spironolactone, eplerenone) may therefore account for the clinical benefit of these compounds in obese and diabetic patients with cardiovascular disease. This review will highlight recent evidence supporting the hypothesis that aldosterone and MR signaling represent an ideal candidate pathway linking early promoters of diabetes, especially overnutrition and obesity, to vascular insulin resistance, dysfunction, and disease.


Pathomechanisms of ALS8: altered autophagy and defective RNA binding protein (RBP) homeostasis due to the VAPB P56S mutation.
Mutations in RNA binding proteins (RBPs) and in genes regulating autophagy are frequent causes of familial amyotrophic lateral sclerosis (fALS). The P56S mutation in vesicle-associated membrane protein-associated protein B (VAPB) leads to fALS (ALS8) and spinal muscular atrophy (SMA). While VAPB is primarily involved in the unfolded protein response (UPR), vesicular trafficking and in initial steps of the autophagy pathway, the effect of mutant P56S-VAPB on autophagy regulation in connection with RBP homeostasis has not been explored yet. Examining the muscle biopsy of our index ALS8 patient of European origin revealed globular accumulations of VAPB aggregates co-localised with autophagy markers LC3 and p62 in partially atrophic and atrophic muscle fibres. In line with this skin fibroblasts obtained from the same patient showed accumulation of P56S-VAPB aggregates together with LC3 and p62. Detailed investigations of autophagic flux in cell culture models revealed that P56S-VAPB alters both initial and late steps of the autophagy pathway. Accordingly, electron microscopy complemented with live cell imaging highlighted the impaired fusion of accumulated autophagosomes with lysosomes in cells expressing P56S-VAPB. Consistent with these observations, neuropathological studies of brain and spinal cord of P56S-VAPB transgenic mice revealed signs of neurodegeneration associated with altered protein quality control and defective autophagy. Autophagy and RBP homeostasis are interdependent, as demonstrated by the cytoplasmic mis-localisation of several RBPs including pTDP-43, FUS, Matrin 3 which often sequestered with P56S-VAPB aggregates both in cell culture and in the muscle biopsy of the ALS8 patient. Further confirming the notion that aggregation of the RBPs proceeds through the stress granule (SG) pathway, we found persistent G3BP- and TIAR1-positive SGs in P56S-VAPB expressing cells as well as in the ALS8 patient muscle biopsy. We conclude that P56S-VAPB-ALS8 involves a cohesive pathomechanism of aberrant RBP homeostasis together with dysfunctional autophagy.


Novel anticancer agents in clinical development.
There are currently more than 100 agents officially approved for the treatment of cancer world-wide. However, the most common epithelial cancers, which cause greater than 75% of cancer deaths, remain incurable. Most therapeutic agents have been developed empirically by testing large numbers of chemicals on rapidly growing transplantable rodent tumors, and more recently, human tumor xenografts. This approach has predominantly identified DNA-active drugs, which have limited efficacy and considerable toxicity. Novel agents, which selectively target aberrant elements in neoplastic cells and their microenvironment, are needed to improve the cure rates of epithelial malignancies. In recent years, advances in the understanding of molecular genetics and tumor biology have elucidated the molecular pathways implicated in the pathogenesis and progression of cancers and resulted in the discovery of a variety of novel molecular targets for therapeutic intervention. These targets can be conceptualized as supportive vessels, connective tissues, and signaling elements. Agents directed against these targets are those that interfere with signal transduction pathways, cell cycle regulation, and apoptosis (signals), malignant angiogenesis (vessels) and the tumor stroma (connective tissue). As anti-cancer therapeutics with distinct targeting capabilities against malignant cells become available for clinical evaluations, prioritization of these therapies for efficient allotment of clinical trial resources, identification of patients whose malignancies most likely express the molecular constituents resembling the true target, and derivation of relevant endpoints for both screening and assessment of clinical relevance will be critical to their ultimate development and success. This review will highlight promising rationally designed, target-based agents in clinical development, as well as the unique challenges involved in their successful development.


Early detection of subclinical visual damage after blast-mediated TBI enables prevention of chronic visual deficit by treatment with P7C3-S243.
Traumatic brain injury (TBI) frequently leads to chronic visual dysfunction. The purpose of this study was to investigate the effect of TBI on retinal ganglion cells (RGCs), and to test whether treatment with the novel neuroprotective compound P7C3-S243 could prevent in vivo functional deficits in the visual system. Blast-mediated TBI was modeled using an enclosed over-pressure blast chamber. The RGC physiology was evaluated using a multielectrode array and pattern electroretinogram (PERG). Histological analysis of RGC dendritic field and cell number were evaluated at the end of the study. Visual outcome measures also were evaluated based on treatment of mice with P7C3-S243 or vehicle control. We show that deficits in neutral position PERG after blast-mediated TBI occur in a temporally bimodal fashion, with temporary recovery 4 weeks after injury followed by chronically persistent dysfunction 12 weeks later. This later time point is associated with development of dendritic abnormalities and irreversible death of RGCs. We also demonstrate that ongoing pathologic processes during the temporary recovery latent period (including abnormalities of RGC physiology) lead to future dysfunction of the visual system. We report that modification of PERG to provocative postural tilt testing elicits changes in PERG measurements that correlate with a key in vitro measures of damage: the spontaneous and light-evoked activity of RGCs. Treatment with P7C3-S243 immediately after injury and throughout the temporary recovery latent period protects mice from developing chronic visual system dysfunction. Provocative PERG testing serves as a noninvasive test in the living organism to identify early damage to the visual system, which may reflect corresponding damage in the brain that is not otherwise detectable by noninvasive means. This provides the basis for developing an earlier diagnostic test to identify patients at risk for developing chronic CNS and visual system damage after TBI at an earlier stage when treatments may be more effective in preventing these sequelae. In addition, treatment with the neuroprotective agent P7C3-S243 after TBI protects from visual system dysfunction after TBI.


Challenges and Opportunities for Biological Mass Spectrometry Core Facilities in the Developing World.
The developing world is seeing rapid growth in the availability of biological mass spectrometry (MS), particularly through core facilities. As proteomics and metabolomics becomes locally feasible for investigators in these nations, application areas associated with high burden in these nations, such as infectious disease, will see greatly increased research output. This article evaluates the rapid growth of MS in South Africa (currently approaching 20 laboratories) as a model for establishing MS core facilities in other nations of the developing world. Facilities should emphasize new services rather than new instruments. The reduction of the delays associated with reagent and other supply acquisition would benefit both facilities and the users who make use of their services. Instrument maintenance and repair, often mediated by an in-country business for an international vendor, is also likely to operate on a slower schedule than in the wealthiest nations. A key challenge to facilities in the developing world is educating potential facility users in how best to design experiments for proteomics and metabolomics, what reagents are most likely to introduce problematic artifacts, and how to interpret results from the facility. Here, we summarize the experience of 6 different institutions to raise the level of biological MS available to researchers in South Africa.


Extracellular proteases and their inhibitors in genetic diseases of the central nervous system.
Cumulative evidence has shown that a delicate balance between serine proteases and their inhibitors is crucial for normal functioning of several biological pathways. The importance of proteases and their inhibitors is well documented in several human diseases. Among them, the best documented are hemophilia B, a genetic deficiency of the serine protease coagulation factor IX and serpinophathies. Alpha-1-antitrypsin deficiency (MIM 107400), is associated with early-onset emphysema and liver disease, while hereditary angioedema (HANE; MIM 106100) is caused by mutations in the C1 inhibitor, a serpin involved in the regulation of the complement cascade. Recently, two human genetic diseases of the central nervous system have been related to mutations in components of extracellular proteolytic systems. Here, we review the recent advances in this field.


Measurement of the charge asymmetry in top quark pair production in pp collisions at [Formula: see text] using the ATLAS detector.
A measurement of the top-antitop production charge asymmetry AC is presented using data corresponding to an integrated luminosity of 1.04 fb-1 of pp collisions at [Formula: see text] TeV collected by the ATLAS detector at the LHC. Events are selected with a single lepton (electron or muon), missing transverse momentum and at least four jets of which at least one jet is identified as coming from a b-quark. A kinematic fit is used to reconstruct the [Formula: see text] event topology. After background subtraction, a Bayesian unfolding procedure is performed to correct for acceptance and detector effects. The measured value of AC is [Formula: see text], consistent with the prediction from the MC@NLO Monte Carlo generator of AC =0.006±0.002. Measurements of AC in two ranges of invariant mass of the top-antitop pair are also shown.


Cell biological changes of outer cortical root cells in early determinate nodulation.
In the symbiosis of leguminous plants and Rhizobium bacteria, nodule primordia develop in the root cortex. This can be either in the inner cortex (indeterminate-type of nodulation) or outer cortex (determinate-type of nodulation), depending upon the host plant. We studied and compared early nodulation stages in common bean (Phaseolus vulgaris) and Lotus japonicus, both known as determinate-type nodulation plants. Special attention was paid to the occurrence of cytoplasmic bridges, the influence of rhizobial Nod factors (lipochitin oligosaccharides [LCOs]) on this phenomenon, and sensitivity of the nodulation process to ethylene. Our results show that i) both plant species form initially broad, matrix-rich infection threads; ii) cytoplasmic bridges occur in L. japonicus but not in bean; iii) formation of these bridges is induced by rhizobial LCOs; iv) formation of primordia starts in L. japonicus in the middle root cortex and in bean in the outer root cortex; and v) in the presence of the ethylene-biosynthesis inhibitor aminoethoxyvinylglycine (AVG), nodulation of L. japonicus is stimulated when the roots are grown in the light, which is consistent with the role of cytoplasmic bridges during nodulation of L. japonicus.


Influence of hemoglobin levels on inpatient rehabilitation outcomes after total knee arthroplasty.
This retrospective study examined the influence of hemoglobin (Hb) on the outcomes of 184 acute inpatient rehabilitation patients admitted to a single university-based inpatient rehabilitation facility after primary total knee arthroplasty between 2001 and 2003. Patient function was measured using the Functional Independence Measure (FIM) instrument. Average length of stay was 9.4 days. Total FIM score increased from 81.5 to 110.8. Mean baseline Hb was 10.5 g/dL. Multivariate analyses demonstrated that a higher Hb at baseline was associated with significantly shorter length of stay (P = .004) and greater FIM efficiency (change in total FIM score/length of stay) (P = .04). Hemoglobin is associated with rehabilitation outcomes after total knee arthroplasty; additional research into the influence of blood management strategies on rehabilitation outcomes is warranted.


Bariatric surgery for severely obese adolescents.
A 1991 National Institutes of Health Consensus Conference concluded that severely obese adults could be eligible for bariatric surgery if they had a body mass index (BMI) > or =35 kg/m(2) with or > or =40 kg/m(2) without obesity comorbidity. It was thought at that time that there were inadequate data to support bariatric surgery in severely obese adolescents. An estimated 25% of children in the United States are obese, a number that has doubled over a 30-year period. Very little information has been published on the subject of obesity surgery in adolescents. Therefore we reviewed our 20-year database on bariatric surgery in adolescents. Severely obese adolescents, ranging from 12 to less than 18 years of age, were considered eligible for bariatric surgery according to the National Institutes of Health adult criteria. Gastroplasty was the procedure of choice in the initial 3 years of the study followed by gastric bypass, which was found to be significantly more effective for weight loss in adults. Distal gastric bypass (D-GBP) was used in extremely obese patients (BMI > or =60 kg/m(2)) before 1992 and long-limb gastric bypass (LL-GBP) was used for superobese patients (BMI > or =50 kg/m(2)) after 1992. Laparoscopic gastric bypass was used after 2000. Thirty-three adolescents (27 white, 6 black; 19 females, 14 males) underwent the following bariatric operations between 1981 and June 2001: horizontal gastroplasty in one, vertical banded gastroplasty in two, standard gastric bypass in 17 (2 laparoscopic), LL-GBP in 10, and D-GBP in three. Mean BMI was 52 +/- 11 kg/m(2) (range 38 to 91 kg/m(2)), and mean age was 16 +/- 1 years (range 12.4 to 17.9 years). Preoperative comorbid conditions included the following: type II diabetes mellitus in two patients, hypertension in 11, pseudotumor cerebri in three, gastroesophageal reflux in five, sleep apnea in six, urinary incontinence in two, polycystic ovary syndrome in one, asthma in one, and degenerative joint disease in 11. There were no operative deaths or anastomotic leaks. Early complications included pulmonary embolism in one patient, major wound infection in one, minor wound infections in four, stomal stenoses (endoscopically dilated) in three, and marginal ulcers (medically treated) in four. Late complications included small bowel obstruction in one and incisional hernias in six patients. There were two late sudden deaths (2 years and 6 years postoperatively), but these were unlikely to have been caused by the bariatric surgical procedure. Revision procedures included one D-GBP to gastric bypass for malnutrition and one gastric bypass to LL-GBP for inadequate weight loss. Regain of most or all of the lost weight was seen in five patients at 5 to 10 years after surgery; however, significant weight loss was maintained in the remaining patients for up to 14 years after surgery. Comorbid conditions resolved at 1 year with the exception of hypertension in two patients, gastroesophageal reflux in two, and degenerative joint disease in seven. Self-image was greatly enhanced; eight patients have married and have children, five patients have completed college, and one patient is currently in college. Severe obesity is increasing rapidly in adolescents and is associated with significant comorbidity and social stigmatization. Bariatric surgery in adolescents is safe and is associated with significant weight loss, correction of obesity comorbidity, and improved self-image and socialization. These data strongly support obesity surgery for those unfortunate individuals who may have difficulty obtaining insurance coverage based on the 1991 National Institutes of Health Consensus Conference statement.


DNA micropatterning on polycrystalline diamond via one-step direct amination.
We report a novel method of one-step direct amination on polycrystalline diamond to produce functionalized surfaces for DNA micropatterning by photolithography. Polycrystalline diamond was exposed to UV irradiation in ammonia gas to generate amine groups directly. After patterning, optical microscopy confirmed that micropatterns covered with an Au mask were regular in size and shape. The regions outside the micropatterns were passivated with fluorine termination by C3F8 plasma, and the chemical changes on the two different surfaces--the amine groups inside the patterned regions by one-step direct amination and fluorine termination outside the patterned regions--were characterized by spatially resolved X-ray photoelectron spectroscopy (XPS). The patterned areas terminated with active amine groups were then immobilized with probe DNA via a bifunctional molecule. The sequence specificity was conducted by hybridizing fluorescently labeled target DNA to both complementary and noncomplementary probe DNA attached inside the micropatterns. The fluorescence micropatterns observed by epifluorescence microscopy corresponded to those imaged by optical microscopy. DNA hybridization and denaturation experiments on a DNA-modified diamond show that the diamond surfaces reveal superior stability. The influence of a different amination time on fluorescence intensity was compared. Different terminations as passivated layers were investigated, and as a result, fluorine termination points to the greatest signal-to-noise ratio.


The Axin1 scaffold protein promotes formation of a degradation complex for c-Myc.
Expression of the c-Myc proto-oncoprotein is tightly regulated in normal cells. Phosphorylation at two conserved residues, threonine58 (T58) and serine62 (S62), regulates c-Myc protein stability. In cancer cells, c-Myc can become aberrantly stabilized associated with altered T58 and S62 phosphorylation. A complex signalling cascade involving GSK3beta kinase, the Pin1 prolyl isomerase, and the PP2A-B56alpha phosphatase controls phosphorylation at these sites. We report here a novel role for the tumour suppressor scaffold protein Axin1 in facilitating the formation of a degradation complex for c-Myc containing GSK3beta, Pin1, and PP2A-B56alpha. Although knockdown of Axin1 decreases the association of c-Myc with these proteins, reduces T58 and enhances S62 phosphorylation, and increases c-Myc stability, acute expression of Axin1 reduces c-Myc levels and suppresses c-Myc transcriptional activity. Moreover, the regulation of c-Myc by Axin1 is impaired in several tested cancer cell lines with known stabilization of c-Myc or loss of Axin1. This study provides critical insight into the regulation of c-Myc expression, how this can be disrupted in three cancer types, and adds to our knowledge of the tumour suppressor activity of Axin1.


Pathogenesis of occult chronic hepatitis B virus infection.
Occult hepatitis B infection (OBI) is characterized by hepatitis B virus (HBV) DNA in serum in the absence of hepatitis B surface antigen (HBsAg) presenting HBsAg-negative and anti-HBc positive serological patterns. Occult HBV status is associated in some cases with mutant viruses undetectable by HBsAg assays; but more frequently it is due to a strong suppression of viral replication and gene expression. OBI is an entity with world-wide diffusion. The failure to detect HBsAg, despite the persistence of the viral DNA, is due in most cases to the strong suppression of viral replication and gene expression that characterizes this "occult" HBV infection; although the mechanisms responsible for suppression of HBV are not well understood. The majority of OBI cases are secondary to overt HBV infection and represent a residual low viremia level suppressed by a strong immune response together with histological derangements which occurred during acute or chronic HBV infection. Much evidence suggests that it can favour the progression of liver fibrosis and the development of hepatocellular carcinoma.


Bedside wireless lung ultrasound for the evaluation of COVID-19 lung injury in senior nursing home residents.
Lung Ultrasound (LUS) is regarded to be potentially useful to diagnose lung injury in older adults living in nursing homes with suspected COVID-19 pneumonia. We aimed at evaluating presence lung injury among senior nursing home residents by LUS performed with portable wireless scanner echography. The study population consisted of 150 residents with a mean age of 88 years (85% female) residing in 12 nursing homes in Northern Italy. Subjects had to have a history of recent onset of symptoms compatible with COVID-19 pneumonia or have been exposed to the contagion of patients carrying the disease. COVID-19 testing was performed with SARS-CoV-2 nasal-pharyngeal (NP) swabs. Positive subjects to LUS scanning were considered those with non-coascelent B-lines in >3 zones, coalescent B-lines in >3 zones and with iperdensed patchy non-consolidated lungs. Sixty-three percent had positive NP testing and 65% had LUS signs of pulmonary injury. LUS had a sensitivity of 79% in predicting positive NP testing. Sixteen percent of residents tested negative for SARSCoV-2 carried the signs of COVID-19 lung injury at LUS. There were 92 patients (61%) with current or recent symptoms.Positivity to LUS scanning was reported in 73% of residents with symptoms, while it was 53% in those without (P=0.016). A positive NP testing was observed in 66% of residents with symptoms and in 57% of those without (P=0.27). We conclude that assessment of LUS by portable wireless scanner echography can be profitability utilized to diagnose lung injury among senior nursing home residents with or without symptoms compatible with COVID-19 pneumonia.


Increased adrenaline, beta-adrenoreceptor stimulation and phospholipid methylation in pineal gland of spontaneously hypertensive rats.
1. In pineal glands of adult spontaneously hypertensive rats (Okamoto strain), dopamine levels increased 34%, adrenaline concentrations increased 152% and noradrenaline levels decreased by 26%, when compared with Wistar-Kyoto normotensive controls. 2. These results are consistent with the hypothesis of an increased uptake of peripheral adrenaline resulting in increased release of noradrenaline from pineal sympathetic nerves in hypertensive rats. 3. The activity of pineal N-acetyltransferase was increased fourfold in hypertensive rats, indicating increased stimulation of pineal beta-adrenoreceptors. 4. Methylation of pineal membrane phospholipids was also increased (100-320%) in hypertensive rats. 5. These results indicate a correlation in vivo between increased beta-adrenoreceptor stimulation and increased methylation of membrane phospholipids in the rat pineal gland, which could result in changes in membrane fluidity and affect the coupling of the beta-adrenoreceptors to the adenylate cyclase.


Differential regulation of human, antigen-specific Th1 and Th2 responses by the B-7 homologues, CD80 and CD86.
A selectivity of B7.1 (CD80) for promoting Th1 responses and B7.2 (CD86) for promoting Th2 responses in the murine system has recently been suggested. The present study explores this hypothesis, using human PBMCs and antigen-specific Th1 and Th2 clones. Proliferative responses of peripheral blood mononuclear cells (PBMCs) from ragweed-allergic, tetanus toxoid-immunized individuals were downregulated by treatment with anti-CD86 in ragweed- and tetanus toxoid-driven cultures (% Inhibition = 55 +/- 4 and 61 +/- 12, respectively; P < 0.03 relative to untreated cultures). Gene expression in PBMCs for interleukin (IL)-4, IL-5, and interferon gamma (IFNgamma), assessed by reverse-transcriptase polymerase chain reaction, was also downregulated by treatment with anti-CD86 in both the ragweed- and tetanus toxoid-driven systems. Neither independent efficacy nor synergy with anti-CD86 was apparent with anti-CD80 treatment; two different anti-CD80 blocking antibodies yielded identical results. Conversely, antigen-specific Th1 and Th2 clones were insensitive to treatment with either anti-CD80, anti-CD86, or a combination of the two. Unaffected parameters included proliferative response (P < 0.14 and 0.33, respectively, for Th1 and Th2), proinflammatory cytokine gene expression, and cytokine protein secretion into culture supernatants (P < 0.44 and 0.16, respectively, for IL-4 and IFNgamma). We conclude that CD86 is the primary B7 signaling homologue in human PBMC responses, and that second signal pathways through the B7 homologues have no effect on phenotypically differentiated T helper cells in humans.


Homogeneous Distribution of Polymerizable Coumarin Dyes for Active Few Mode POF.
For most kinds of active polymer optical fibers, a homogeneous distribution of dye molecules over the entire fiber length and cross section is required. In this study, chemical bonding of dyes to poly(methyl methacrylate) (PMMA) by copolymerization is achieved within the polymerization process instead of dissolving the dyes in the monomers. In combination with an improved fabrication mechanism, this leads to homogeneous dye distribution within the preforms. A method for proving the integration of the dyes into the polymer chains has been developed using high-performance liquid chromatography (HPLC) and size exclusion chromatography (SEC). Prestructured core-cladding preforms with dye-doped poly(cylohexyl methacrylate-co-methyl methacrylate)-core have been prepared with the Teflon string technique and were heat-drawn to few mode fibers.


Bacteriophage control of Pseudomonas plecoglossicida infection in ayu Plecoglossus altivelis.
Two previously isolated phages were used to examine the therapeutic effects against Pseudomonas plecoglossicida infection in ayu Plecoglossus altivelis. Phage PPp-W4 (Podoviridae) inhibited the in vitro growth of P. plecoglossicida more effectively than Phage PPpW-3 (Myoviridae), and a mixture (PPpW-3/W-4) of the 2 phages exhibited the highest inhibitory activity. In phage therapy experiments, ayu were fed P. plecoglossicida-impregnated feed (10(7) CFU fish(-1)) and then fed phage-impregnated feed (10(7) PFU fish(-1)). Mortalities of fish receiving PPpW-3, PPpW-4, PPpW-3/W-4, and a control fish receiving no phages were 53.3, 40.0, 20.0 and 93.3%, respectively. Phage (PPpW-3/W-4)-receiving fish also showed high protection against water-borne infection with P. plecoglossicida. In a field trial, when phage (PPpW-3/W-4)-impregnated feed was administered to ayu in a pond where the disease occurred naturally, daily mortality of fish decreased at a constant level (5% d(-1)) to one-third after a 2 wk period. The causal relationship of phages in this phenomenon was verified by the long-lasting appearance of administered phages in the kidneys of the fish, and a disappearance of P. plecoglossicida from apparently healthy fish. Neither phage-resistant organisms nor phage-neutralizing antibodies were detected in diseased fish or apparently healthy fish, respectively. These results indicate the potential for phage control of the disease.


CaMKIIα underlies spontaneous and evoked pain behaviors in Berkeley sickle cell transgenic mice.
Pain is one of the most challenging and stressful conditions to patients with sickle cell disease (SCD) and their clinicians. Patients with SCD start experiencing pain as early as 3 months old and continue having it throughout their lives. Although many aspects of the disease are well understood, little progress has been made in understanding and treating pain in SCD. This study aimed to investigate the functional involvement of Ca/calmodulin-dependent protein kinase II (CaMKIIα) in the persistent and refractory pain associated with SCD. We found that nonevoked ongoing pain as well as evoked hypersensitivity to mechanical and thermal stimuli were present in Berkeley sickle cell transgenic mice (BERK mice), but not nonsickle control littermates. Prominent activation of CaMKIIα was observed in the dorsal root ganglia and spinal cord dorsal horn region of BERK mice. Intrathecal administration of KN93, a selective inhibitor of CaMKII, significantly attenuated mechanical allodynia and heat hyperalgesia in BERK mice. Meanwhile, spinal inhibition of CaMKII elicited conditioned place preference in the BERK mice, indicating the contribution of CaMKII in the ongoing spontaneous pain of SCD. We further targeted CaMKIIα by siRNA knockdown. Both evoked pain and ongoing spontaneous pain were effectively attenuated in BERK mice. These findings elucidated, for the first time, an essential role of CaMKIIα as a cellular mechanism in the development and maintenance of spontaneous and evoked pain in SCD, which can potentially offer new targets for pharmacological intervention of pain in SCD.


Modulation of laryngeal responses to superior laryngeal nerve stimulation by volitional swallowing in awake humans.
Laryngeal sensori-motor closure reflexes are important for the protection of the airway and prevent the entry of foreign substances into the trachea and lungs. The purpose of this study was to determine how such reflexes might be modulated during volitional swallowing in awake humans, when persons are at risk of entry of food or liquids into the airway. The frequency and the amplitude of laryngeal adductor responses evoked by electrical stimulation of the internal branch of the superior laryngeal nerve (ISLN) were studied during different phases of volitional swallowing. Subjects swallowed water on command while electrical stimuli were presented to the ISLN at various intervals from 500 ms to 5 s following the command. Laryngeal adductor responses to unilateral ISLN stimulation were recorded bilaterally in the thyroarytenoid muscles using hooked wire electrodes. Early ipsilateral R1 responses occurred at 17 ms, and later bilateral R2 began around 65 ms. The muscle responses to stimuli occurring during expiration without swallowing were quantified as control trials. Responses to stimulation presented before swallowing, during the swallow, within 3 s after swallowing, and between 3 and 5 s after a swallow were measured. The frequency and amplitude of three responses (ipsilateral R1 and bilateral R2) relative to the control responses were compared across the different phases relative to the occurrence of swallowing. Results demonstrated that a reduction occurred in both the frequency and amplitude of the later bilateral R2 laryngeal responses to electrical stimulation for up to 3 s after swallowing (P = 0.005). The amplitude and frequency of ipsilateral R1 laryngeal responses, however, did not show a significant main effect following the swallow (P = 0.28), although there was a significant time by measure interaction (P = 0.006) related to reduced R1 response amplitude up to 3 s after swallowing (P = 0.021). Therefore, the more rapid and shorter unilateral R1 responses continued to provide some, albeit reduced, laryngeal protective functions after swallowing, whereas the later bilateral R2 responses were suppressed both in occurrence and amplitude for up to 3 s after swallowing. The results suggest that R2 laryngeal adductor responses are suppressed following swallowing when residues may remain in the laryngeal vestibule putting persons at increased risk for the entry of foreign substances into the airway.


Deformable segmentation of 3D MR prostate images via distributed discriminative dictionary and ensemble learning.
Automatic prostate segmentation from MR images is an important task in various clinical applications such as prostate cancer staging and MR-guided radiotherapy planning. However, the large appearance and shape variations of the prostate in MR images make the segmentation problem difficult to solve. Traditional Active Shape/Appearance Model (ASM/AAM) has limited accuracy on this problem, since its basic assumption, i.e., both shape and appearance of the targeted organ follow Gaussian distributions, is invalid in prostate MR images. To this end, the authors propose a sparse dictionary learning method to model the image appearance in a nonparametric fashion and further integrate the appearance model into a deformable segmentation framework for prostate MR segmentation. To drive the deformable model for prostate segmentation, the authors propose nonparametric appearance and shape models. The nonparametric appearance model is based on a novel dictionary learning method, namely distributed discriminative dictionary (DDD) learning, which is able to capture fine distinctions in image appearance. To increase the differential power of traditional dictionary-based classification methods, the authors' DDD learning approach takes three strategies. First, two dictionaries for prostate and nonprostate tissues are built, respectively, using the discriminative features obtained from minimum redundancy maximum relevance feature selection. Second, linear discriminant analysis is employed as a linear classifier to boost the optimal separation between prostate and nonprostate tissues, based on the representation residuals from sparse representation. Third, to enhance the robustness of the authors' classification method, multiple local dictionaries are learned for local regions along the prostate boundary (each with small appearance variations), instead of learning one global classifier for the entire prostate. These discriminative dictionaries are located on different patches of the prostate surface and trained to adaptively capture the appearance in different prostate zones, thus achieving better local tissue differentiation. For each local region, multiple classifiers are trained based on the randomly selected samples and finally assembled by a specific fusion method. In addition to this nonparametric appearance model, a prostate shape model is learned from the shape statistics using a novel approach, sparse shape composition, which can model nonGaussian distributions of shape variation and regularize the 3D mesh deformation by constraining it within the observed shape subspace. The proposed method has been evaluated on two datasets consisting of T2-weighted MR prostate images. For the first (internal) dataset, the classification effectiveness of the authors' improved dictionary learning has been validated by comparing it with three other variants of traditional dictionary learning methods. The experimental results show that the authors' method yields a Dice Ratio of 89.1% compared to the manual segmentation, which is more accurate than the three state-of-the-art MR prostate segmentation methods under comparison. For the second dataset, the MICCAI 2012 challenge dataset, the authors' proposed method yields a Dice Ratio of 87.4%, which also achieves better segmentation accuracy than other methods under comparison. A new magnetic resonance image prostate segmentation method is proposed based on the combination of deformable model and dictionary learning methods, which achieves more accurate segmentation performance on prostate T2 MR images.


Mortality among shipyard Coast Guard workers: a retrospective cohort study.
The mortality experience of 4702 (4413 men and 289 women) civilian workers in a US Coast Guard shipyard was evaluated. All workers employed at the shipyard between 1 January 1950 and 31 December 1964 were included in the study and were followed through 31 December 2001 for vital status. Detailed shipyard and lifetime work histories found in the shipyard personnel records and job descriptions were evaluated. Workers were classified as likely exposed to any potential hazardous substances. In addition, 20 job groups were created on likely similar exposures. Standardised mortality ratios (SMRs) were calculated based on the general population of the state and adjusted for age, calendar period, sex and race. The follow-up was successful for 93.3% of the workers. Among all men employed in the shipyard, there was an excess of mortality from all causes of death (SMR 1.08; 95% CI 1.04 to 1.12), respiratory cancers (SMR 1.29; 95% CI 1.15 to 1.43), lung cancer (SMR 1.26; 95% CI 1.12 to 1.41), mesothelioma (SMR 5.07; 95% CI 1.85 to 11.03) and emphysema (SMR 1.44; 95% CI 1.01 to 1.99) and a decrease for cardiovascular diseases (OR 0.95; 95% CI 0.90 to 1.00), vascular lesions of the central nervous system (SMR 0.80; 95% CI 0.67 to 0.96), cirrhosis of the liver (SMR 0.38; 95% CI 0.25 to 0.57) and external causes of death (SMR 0.55; 95% CI 0.44 to 0.68). A similar pattern was observed for the men classified as exposed. No increasing trend of mortality was found with duration of employment in the shipyard, with the exception of mesothelioma (SMRs of 4.23 and 6.27 for <10 years and > or =10 years, respectively). In occupations with at least three cases and with an SMR of > or =1.3, the authors observed a significantly elevated mortality for lung cancer among machinists (SMR 1.60; 95% CI 1.08 to 2.29) and shipfitters, welders and cutters (SMR 1.34; 95% CI 1.07 to 1.65) and for oral and nasopharyngeal cancers among wood workers (SMR 6.20; 95% CI 2.27 to 13.50). Employment in this Coast Guard shipyard revealed a small but significant excess mortality from all causes, lung cancer and mesothelioma, most of which is probably related to asbestos exposure.


Bioluminescence imaging of Toxoplasma gondii infection in living mice reveals dramatic differences between strains.
We examined the in vivo growth, dissemination, and reactivation of strains of the protozoan parasite Toxoplasma gondii using a bioluminescence-based imaging system. Two T. gondii strains, one with a highly virulent disease phenotype in mice (S23) and the other with a 1,000-fold-lower virulence phenotype (S22), were engineered to stably express the light-emitting protein luciferase. One clone of each wild-type strain was isolated, and the two clones (S23-luc7 and S22-luc2) were found to express similar levels of luciferase. Mice were infected intraperitoneally with S23-luc7 (50 or 5 parasites) or S22-luc2 (500, 50, or 5 parasites), and the progress of the infections was examined noninvasively following injection of the substrate for luciferase, d-luciferin. In mice infected with 50 S23-luc7 parasites, the parasites grew exponentially within the peritoneal cavity (as measured by light emitted from luciferase-expressing parasites) during days 1 to 10 p.i., and this proliferation continued until there was severe disease. In mice infected with 500 S22-luc2 parasites, the parasites proliferated in a fashion similar to the S23-luc7 proliferation during days 1 to 6, but this was followed by a precipitous drop in the signal to levels below the limit of detection. Using this technique, we were also able to observe the process of reactivation of T. gondii in chronically infected mice. After treatment with dexamethasone, we detected reactivation of toxoplasmosis in mice infected with S23-luc7 and S22-luc2. During reactivation, growth of S23-luc7 was initially detected primarily in the head and neck area, while in S22-luc2-infected mice the parasites were detected primarily in the abdomen. This method has great potential for identifying important differences in the dissemination and growth of different T. gondii strains, especially strains with dramatically different disease outcomes.


Expansion of adipose tissue mesenchymal stromal progenitors in serum-free medium supplemented with virally inactivated allogeneic human platelet lysate.
Single-donor or pooled platelet lysates (PL) can substitute for fetal bovine serum (FBS) for mesenchymal stromal cell (MSC) expansion. However, for clinical applications of MSCs, the use of virally inactivated PL would be desirable. Recently, we have developed a solvent/detergent (S/D)-treated human PL preparation (S/D-PL) rich in growth factors. The capacity to use this virally inactivated preparation for MSC expansion needs to be evaluated. Platelet concentrates were treated by S/D (1% tri-n-butyl phosphate and 1% Triton X-45), extracted by oil, purified by C18 hydrophobic interaction chromatography, and sterile filtered. S/D-PL was compared to FBS as a medium supplement (10% vol/vol) for isolating, maintaining, and expanding adipose tissue-derived MSCs (AT-MSCs). Cell morphology; proliferation kinetics; immunophenotype; differentiation capacity toward the chondrogenic, osteogenic, and osteogenic lineages; and cytokine antibody array were assessed. AT-MSCs had a typical spindle morphology and proliferated in S/D-PL at least as well as in FBS. Immunophenotype at Passage 7 was characteristic of MSCs and similar for both culture conditions. Differentiation capacity into the three lineages was maintained and chondrogenesis was enhanced by S/D-PL. In a 120 human cytokine antibody array analysis, 73 cytokines were detected in S/D-PL, including 22 with a concentration higher than in FBS. S/D-PL is an alternative to FBS for AT-MSC maintenance and expansion, does not compromise the differentiation capacity nor the immunophenotype, and may accelerate chondrogenesis. S/D-PL protocols for MSC clinical scale-up may represent a major step toward challenging new use in stem cell therapies.


Serum prostate-specific antigen in a community-based population of healthy men. Establishment of age-specific reference ranges.
To define the characteristics of serum prostate-specific antigen (PSA) in a population of healthy men without clinically evident prostate cancer, but who are at risk for developing the malignancy. The influence of patient age and prostatic size on the serum PSA concentration was assessed in order to use PSA more appropriately to detect clinically significant prostate cancer at an early, potentially curable stage. Prospective, community-based study. Between December 1989 and March 1991, 2119 healthy men aged 40 to 79 years from Olmsted County, Minnesota, were entered into a prospective study to assess the natural history of benign prostatic hyperplasia. Of these, 537 (25%) were randomly chosen to participate in a detailed clinical examination that included a serum PSA determination (Tandem-R PSA assay), digital rectal examination, and transrectal ultrasonography. Four hundred seventy-one (88%) completed the prostatic evaluation and had no evidence of prostate cancer by any of these three diagnostic tests; these men formed the study population on which all analyses were performed. Serum PSA concentration, prostatic volume, and PSA density (serum PSA level/prostatic volume) as a function of patient age. The serum PSA concentration is correlated with patient age (r = .43; P < .0001) and prostatic volume (r = .55; P < .0001). Prostatic volume, in turn, is directly correlated with patient age (r = .43; P < .0001), whereas the PSA density value is only weakly correlated with patient age (r = .25; P < .001). For a healthy 60-year-old man with no evidence of prostate cancer, the serum PSA concentration increases by approximately 3.2% per year (0.04 ng/mL per year). The recommended reference range for serum PSA (95th percentile) for men aged 40 to 49 years is 0.0 to 2.5 ng/mL; for 50 to 59 years, 0.0 to 3.5 ng/mL; 60 to 69 years, 0.0 to 4.5 ng/mL; and 70 to 79 years, 0.0 to 6.5 ng/mL. The serum PSA concentration is directly correlated with patient age and prostatic volume, the latter of which also is directly related to age. Thus, rather than rely on a single reference range for men of all age groups, it is more appropriate to have age-specific reference ranges. These age-specific reference ranges have the potential to make serum PSA a more discriminating tumor marker for detecting clinically significant cancers in older men (increasing specificity) and to find more potentially curable cancers in younger men (increasing sensitivity).


The FIFA Women's World Cup in Germany 2011--a practical example for tailoring an event-specific enhanced infectious disease surveillance system.
Mass gatherings require a decision from public health authorities on how to monitor infectious diseases during the event. The appropriate level of enhanced surveillance depends on parameters like the scale of the event (duration, spatial distribution, season), participants' origin, amount of public attention, and baseline disease activity in the host country. For the FIFA Men's World Cup 2006, Germany implemented enhanced surveillance. As the scale of the FIFA Women's World Cup (June 26 - July 17, 2011) was estimated to be substantially smaller in size, visitors and duration, it was not feasible to simply adopt the previously implemented measures. Our aim was therefore to develop a strategy to tailor an event-specific enhanced surveillance for this smaller-scale mass gathering. Based on the enhanced surveillance measures during the Men's Cup, we conducted a needs assessment with the district health authorities in the 9 host cities in March 2011. Specific measures with a majority consent were implemented. After the event, we surveyed the 9 district and their corresponding 7 state health authorities to evaluate the implemented measures. All 9 district health authorities participated in the pre-event needs assessment. The majority of sites consented to moving from weekly to daily (Monday-Friday) notification reporting of routine infectious diseases, receiving regular feedback on those notification reports and summaries of national/international World Cup-relevant epidemiological incidents, e.g. outbreaks in countries of participating teams. In addition, we decided to implement twice-weekly reports of "unusual events" at district and state level. This enhanced system would commence on the first day and continue to one day following the tournament. No World Cup-related infectious disease outbreaks were reported during this time period. Eight of 9 district and 6 of 8 state health authorities participated in the final evaluation. The majority perceived the implemented measures as adequate. Our approach to tailor an event-specific enhanced surveillance concept worked well. Involvement of the participating stakeholders early-on in the planning phase secured ownership of and guaranteed support for the chosen strategy. The enhanced surveillance for this event resulted as a low-level surveillance. However, we included mechanisms for rapid upscaling if the situation would require adaptations.


Cost of treatment in patients with metastatic soft tissue sarcoma who respond favourably to chemotherpy. The SArcoma treatment and Burden of Illness in North America and Europe (SABINE) study.
Treatment of metastatic soft tissue sarcoma (mSTS) commonly includes multiple lines of chemotherapy, until a decline in performance status precludes further treatment. The primary objective of this study was to describe the lifetime healthcare resource utilisation and cost among mSTS patients with favourable response to chemotherapy. SABINE was a multi-centre (n = 25), multi-country (n = 9) retrospective chart review study of mSTS patients with favourable response to chemotherapy following 4 cycles. Healthcare resource utilisation was collected from first line until death or end of follow-up. Costs were analysed by health states (defined by treatment line, chemotherapy use and disease progression) and estimated by multiplying the mean weekly cost per health state by the expected number of weeks spent in each health state. Expected per-patient lifetime medical cost was €65 616 (95% CI: €51 454-€85 003); comprised of IV chemotherapy (31.7%), inpatient care (24.8%), concomitant medication (11.0%), oral chemotherapy (8.9%), outpatient visits (8.8%), radiotherapy (6.3%), hospice (4.0%), imaging (3.7%) and laboratory (0.7%). Weekly costs were 280-330% higher during chemotherapy treatment periods than off-chemotherapy, especially after disease progression. Per-patient costs were highest in the USA and lowest in the Netherlands and UK. The economic burden of mSTS is considerable and the amount of resources devoted to its treatment varies across countries.


A randomised controlled trial of a probiotic 'functional food' in the management of irritable bowel syndrome.
Irritable Bowel Syndrome (IBS) is a common condition characterised by pain, distension and altered bowel habit. Evidence suggests functional foods containing probiotics improve gastrointestinal transit, however, data are limited by short follow-up periods and evaluation in selected populations. A multi-centre, randomized, double blind, controlled trial to evaluate the effect of a probiotic vs non-probiotic dairy product on symptoms in IBS with a constipation element (IBS-Constipation or IBS-Mixed profile). Set in 13 general practices within central England. Individuals meeting the ROME III criteria for IBS, aged 18-65 completed a pre-study diary. Eligible individuals were randomized to consume dairy 'yoghurt' products which either did or did not contain active probiotics twice daily and to complete a daily diary. Primary outcome was subjective global assessment of symptom relief at week 4. Other outcomes comprised, IBS symptom scores, pain, bloating and flatulence levels, stool frequency, stool consistency, ease of bowel movement and quality of life. 179 were randomized (91 active, 88 placebo). 76 (43 active, 33 placebo) completed the study. No significant between group differences existed at 4 weeks (57% active vs 53% placebo, reported adequate relief (p = 0.71)). By week 8, 46% active vs 68% placebo reported adequate relief (p = 0.03). This was sustained at week 12. Significant improvements were reported for most outcomes in all trial participants but improvement did not differ by group. This trial does not provide evidence for effectiveness of a probiotic in IBS, in variance with a body of published literature and review conclusions. Differential drop out may however cloud interpretation of data. UK TRIAL REGISTRATION: ISRCTN78863629.


The challenge of haemoglobinopathies in resource-poor countries.
The haemoglobinopathies, inherited disorders of the structure or synthesis of haemoglobin, are the commonest monogenic diseases. Approximately 80% of the annual births of babies with these conditions occur in low-or middle-income countries, many of which have extremely limited facilities for their control and management. Given that the population size of many of them is growing and, as social and public health facilities improve, increasing numbers of these babies will survive to present for diagnosis and treatment. Hence, the haemoglobinopathies will constitute an increasing global health burden. Hitherto, they have been largely ignored by governments of high-frequency countries and by the international health agencies. However, a start has been made in developing control programmes in some low-income countries and there is already considerable evidence that much can be done to improve the situation by the development of partnerships between groups in richer countries and centres in low-income countries. The natural extension of this approach is the further development of partnerships between countries where expertise in this field has been developed and adjacent countries where no such expertise exists. It is vital that the haematology community of the richer countries becomes involved in programmes of this type while, at the same time, putting pressure on their governments and on international health agencies for support for this work.


Competitive electron transfer and enhanced intersystem crossing in photoexcited covalent TEMPO-perylene-3,4:9,10-bis(dicarboximide) dyads: unusual spin polarization resulting from the radical-triplet interaction.
A stable 2,2,6,6-tetramethyl-1-piperidinyloxyl (TEMPO) radical was covalently attached at its 4-position to the imide nitrogen atom of a perylene-3,4:9,10-bis(dicarboximide) (PDI) to produce TEMPO-PDI, 1, having a well-defined distance and orientation between TEMPO and PDI. Transient optical absorption experiments in toluene following selective photoexcitation of the PDI chromophore in TEMPO-PDI show that enhanced intersystem crossing occurs with tau = 45 +/- 1 ps, resulting in formation of TEMPO-(3*)PDI, while the same experiment in THF shows that the electron-transfer reaction TEMPO-(1*)PDI --> TEMPO(+*)-PDI(-*) occurs with tau = 1.2 +/- 0.2 ps and thus competes effectively with enhanced intersystem crossing. Time-resolved EPR (TREPR) spectroscopy on the photogenerated three-spin system TEMPO-(3*)PDI in toluene at 295 K initially shows a broad signal assigned to spin-polarized (3*)PDI, which thermalizes at longer times and is accompanied by formation of an emissively polarized TEMPO radical. No signals are observed in THF at 295 K. The TREPR spectrum of TEMPO-(3*)PDI at 85 K in toluene shows an emissive/absorptive signal due to TEMPO and a broad triplet signal due to (3*)PDI having a spin polarization pattern characteristic of overpopulation of its T(0) sublevel. This unusual spin polarization pattern does not result from radical pair intersystem crossing because electron transfer does not occur at 85 K. The observed spin polarization of (3*)PDI cannot be readily explained by mechanisms discussed previously, leading us to propose a new spin polarization mechanism, which requires that the radical and attached triplet are in the weak exchange regime.


The Pax gene eyegone facilitates repression of eye development in Tribolium.
The Pax transcription factor gene eyegone (eyg) participates in many developmental processes in Drosophila, including the Notch signaling activated postembryonic growth of the eye primordium, global development of the adult head and the development of the antenna. In contrast to other Pax genes, the functional conservation of eyg in species other than Drosophila has not yet been explored. We investigated the role of eyg during the postembryonic development of the red flour beetle Tribolium castaneum. Our results indicate conserved roles in antennal but not in eye development. Besides segmentation defects in the antenna, Tribolium eyg knockdown animals were characterized by eye enlargement due to the formation of surplus ommatidia at the central anterior edge of the compound eye. This effect resulted from the failure of the developing gena to locally repress retinal differentiation, which underlies the formation of the characteristic anterior notch in the Tribolium eye. Neither varying the induction time point of eyg knockdown nor knocking down components of the Janus kinase/Signal Transducer and Activators of Transcription signaling pathway in combination with eyg reduced eye size like in Drosophila. Taken together, expression and knockdown data suggest that Tribolium eyg serves as a competence factor that facilitates the repression of retinal differentiation in response to an unknown signal produced in the developing gena. At the comparative level, our findings reveal diverged roles of eyg associated with the evolution of different modes of postembryonic head development in endopterygote insects as well as diversified head morphologies in darkling beetles.


Receptor-activated Ca2+ inflow in animal cells: a variety of pathways tailored to meet different intracellular Ca2+ signalling requirements.
Receptor-activated Ca2+ channels (RACCs) play a central role in regulation of the functions of animal cells. Together with voltage-operated Ca2+ channels (VOCCs) and ligand-gated non-selective cation channels, RACCs provide a variety of pathways by which Ca2+ can be delivered to the cytoplasmic space and the endoplasmic reticulum (ER) in order to initiate or maintain specific types of intracellular Ca2+ signal. Store-operated Ca2+ channels (SOCs), which are activated by a decrease in Ca2+ in the ER, are a major subfamily of RACCs. A careful analysis of the available data is required in order to discern the different types of RACCs (differentiated chiefly on the basis of ion selectivity and mechanism of activation) and to properly develop hypotheses for structures and mechanisms of activation. Despite much intensive research, the structures and mechanisms of activation of RACCs are only now beginning to be understood. In considering the physiological functions of the different RACCs, it is useful to consider the specificity for Ca2+ of each type of cation channel and the rate at which Ca2+ flows through a single open channel; the locations of the channels on the plasma membrane (in relation to the ER, cytoskeleton and other intracellular units of structure and function); the Ca2+-responsive enzymes and proteins; and the intracellular buffers and proteins that control the distribution of Ca2+ in the cytoplasmic space. RACCs which are non-selective cation channels can deliver Ca2+ directly to specific regions of the cytoplasmic space, and can also admit Na+, which induces depolarization of the plasma membrane, the opening of VOCCs and the subsequent inflow of Ca2+. SOCs appear to deliver Ca2+ specifically to the ER, thereby maintaining oscillating Ca2+ signals.


Molecular cloning of a pancreatic islet-specific glucose-6-phosphatase catalytic subunit-related protein.
A pancreatic islet-specific glucose-6-phosphatase-related protein (IGRP) was cloned using a subtractive cDNA expression cloning procedure from mouse insulinoma tissue. Two alternatively spliced variants that differed by the presence or absence of a 118-bp exon (exon IV) were detected in normal balb/c mice, diabetic ob/ob mice, and insulinoma tissue. The longer, 1901-bp full-length cDNA encoded a 355-amino acid protein (molecular weight 40,684) structurally related (50% overall identity) to the liver glucose-6-phosphatase and exhibited similar predicted transmembrane topology, conservation of catalytically important residues, and the presence of an endoplasmic reticulum retention signal. The shorter transcript encoded two possible open reading frames (ORFs), neither of which possessed His174, a residue thought to be the phosphoryl acceptor (Pan CJ, Lei KJ, Annabi B, Hemrika W, Chou JY: Transmembrane topology of glucose-6-phosphatase. J Biol Chem 273:6144-6148, 1998). Northern blot and reverse transcription-polymerase chain reaction analysis showed that the mRNA was highly expressed in pancreatic islets and expressed more in beta-cell lines than in an alpha-cell line. It was notably absent in tissues and cell lines of non-islet neuroendocrine origin, and no other major tissue source of the mRNA was found. During development, it was expressed in parallel with insulin mRNA. The mRNA was efficiently translated and glycosylated in an in vitro translation/membrane translocation system and readily transcribed into COS 1, HIT, and CHO cells using cytomegalovirus or Rous sarcoma virus promoters. Whereas the liver glucose-6-phosphatase showed activity in these transfection systems, the IGRP failed to show glucose phosphotransferase or phosphatase activity with p-nitrophenol phosphate, inorganic pyrophosphate, or a range of sugar phosphates hydrolyzed by the liver enzyme. While the metabolic function of the enzyme is not resolved, its remarkable tissue-specific expression warrants further investigation, as does its transcriptional regulation in conditions where glucose responsiveness of the pancreatic islet is altered.


Characterization of lethal inhalational infection with Francisella tularensis in the common marmoset (Callithrix jacchus).
The intracellular Gram-negative pathogen Francisella tularensis is the causative agent of tularaemia and is prevalent in many countries in the northern hemisphere. To determine whether the common marmoset (Callithrix jacchus) would be a suitable non-human primate model of inhalational tularaemia, a pathophysiology study was undertaken. Ten animals were challenged with approximately 10(2) c.f.u. F. tularensis strain SCHU S4 (F. tularensis subsp. tularensis). To look for trends in the infection, pairs of animals were sacrificed at 24 h intervals between 0 and 96 h post-challenge and blood and organs were assessed for bacteriology, pathology and haematological and immunological parameters. The first indication of infection was a raised core temperature at 3 days post-challenge. This coincided with a number of other factors: a rapid increase in the number of bacteria isolated from all organs, more pronounced gross pathology and histopathology, and an increase in the immunological response. As the disease progressed, higher bacterial and cytokine levels were detected. More extensive pathology was observed, with multifocal lesions seen in the lungs, liver and spleen. Disease progression in the common marmoset appears to be consistent with human clinical and pathological features of tularaemia, indicating that this may be a suitable animal model for the investigation of novel medical interventions such as vaccines or therapeutics.


Micro-resonator with metallic mirrors coupled to a bus waveguide.
We demonstrate a micro-resonator based on a channel waveguide terminated with metallic mirrors side coupled to a bus waveguide. Transmission through such a resonant structure implemented in a silicon-on-insulator platform is investigated theoretically and demonstrated experimentally. The resonator is 13.4 μm long, exhibits an unloaded Q-factor of ~2100, and a free spectral range of 21 nm around the wavelength of 1.55 μm.