Effect of a nutrient mixture on matrix metalloproteinase-9 dimers in various human cancer cell lines.
Strong clinical and experimental evidence demonstrates association of elevated levels of matrix metalloproteinase MMP-9 with cancer progression, metastasis and shortened patient survival, as it plays a key role in tumor cell invasion and metastasis by digesting the basement membrane and ECM components. MMP-9 is secreted in both the monomeric and dimeric form. Although there is little research on MMP-9 dimers, some studies have shown the dimer to be associated with more aggressive tumor progression. Our objective was to study the relative secretion patterns of MMP-9 monomer and dimer in a variety of cancer cell lines and the effect of a nutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract on MMP-9 secretion. The cancer cell lines were grown in their respective media, supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 µg/ml) in 24-well tissue culture plates. At near confluence, the cells were treated with NM at 0,10, 50, 100, 500 and 1000 µg/ml. Parallel sets of cultures were treated with PMA (100 ng/ml) for induction of MMP-9. Cell MMP-9 secretion was assayed by gelatinase zymography. MMP-9 dimer secretion patterns of cancer cells fell into different categories. We observed no MMP-9 dimer in prostate DU-145 and PC-3, pancreatic MIA-Pa-Ca2, colon HCT-116, bladder T-24, head and neck FaDu, glioblastoma A-172, T-98 and LN-18 and leukemia HL-60, Jurkat, and Raji cell lines. MMP-dimer secretion only with PMA induction was seen in breast MCF-7 and MDA-MB-231, uterine SK-UT-1, lung A-549, tongue SC-25, melanoma A2058, osteosarcoma U-2OS, rhabdomyosarcoma, fibrosarcoma HT-1080, chondrosarcoma SW-1350 and liposarcoma SW-872. Cervical HeLa and DoTc 2 4510, renal 786-0 and HCC SK-Hep-1 cells exhibited MMP-9 dimer without PMA treatment and increased secretion with PMA treatment. Sarcomas had the highest levels of MMP-9 monomer and dimer with and without PMA among these cancer cell lines. Cervical, uterine and male breast cancer cell lines showed the next highest levels of MMP-9, followed by breast cancer cell lines. Melanoma, renal, lung, head and neck and HCC showed lower levels and prostate, glioblastoma, bladder and leukemia cell lines the lowest. NM showed dose-dependent inhibition of MMP-9 monomer and dimer in all cell lines tested. In conclusion, high MMP-9 and dimer secretion levels correlated with the most aggressive cancer cell lines. NM was effective in inhibiting MMP-9 and dimer secretion in all cell lines tested, suggesting its therapeutic potential as an antimetastatic agent.


Fluid and particle retention in the digestive tract of the addax antelope (Addax nasomaculatus)--adaptations of a grazing desert ruminant.
Retention time of food in the digestive tract is a major aspect describing the digestive physiology of herbivores. Differences in feed retention times have been described for different ruminant feeding types. In this study, a dominantly grazing desert ruminant, the addax (Addax nasomaculatus), was investigated in this respect. Eight animals with a body weight (BW) of 87+/-5.3 kg on an ad libitum grass hay (Chloris gayana) diet were available. Co-EDTA and Cr-mordanted fibers (<2 mm) were used as pulse-dose markers. Mean retention time (MRT) in the digestive tract was calculated from faecal marker excretion. Average daily intake of the addax was found to be 1.7 kg dry matter (DM) or 60+/-8.3 g DM/kg BW(0.75). The MRT of fluid and particles in the reticulo-rumen (MRT(fluid)RR and MRT(particle)RR) were quantified to be 20+/-5.8 and 42+/-7.0 h respectively. When compared to literature data, MRT(fluid)RR was significantly longer than in cattle species, and MRT(particle)RR was significantly longer than in 11 taxa of all feeding types. The ratio of MRT(particle)RR/MRT(fluid)RR (2.3+/-0.5) was found to be within the range described for grazing ruminants. The long retention times found in the addax can be interpreted as an adaptation to a diet including a high proportion of slow fermenting grasses, while the long retention time of the fluid phase can be interpreted as a consequence of water saving mechanisms of the desert-adapted addax with a potentially low water turnover and capacious water storing rumen.


Anti-inflammatory actions of 15-deoxy-delta 12,14-prostaglandin J2 and troglitazone: evidence for heat shock-dependent and -independent inhibition of cytokine-induced inducible nitric oxide synthase expression.
In this study, the anti-inflammatory actions of the peroxisome proliferator-activated receptor (PPAR)-gamma agonists 15-deoxy-delta 12,14-prostaglandin J2 (15-d-delta 12,14-PGJ2) and troglitazone have been examined. Treatment of RAW 264.7 cells and CD-1 mouse peritoneal macrophages with lipopolysaccharide (LPS) + interferon-gamma (IFN-gamma) results in inducible nitric oxide synthase (iNOS), inducible cyclooxygenase (COX-2) and interleukin-1 (IL-1) expression, increased production of nitric oxide, and the release of IL-1. In a concentration-dependent manner, 15-d-delta 12,14-PGJ2 inhibits each of these proinflammatory actions of LPS + IFN-gamma, with half-maximal inhibition at approximately 0.5 microg/ml and complete inhibition at 1-5 microg/ml. The inhibitory actions of 15-d-delta 12,14-PGJ2 on LPS + IFN-gamma-induced inflammatory events are not associated with the inhibition of iNOS enzymatic activity or macrophage cell death, but appear to result from an inhibition of iNOS and IL-1 transcription. In addition, the anti-inflammatory actions of 15-d-delta 12,14-PGJ2 are not limited to peritoneal macrophages, as 15-d-delta 12,14-PGJ2 prevents TNF-alpha + LPS-induced resident islet macrophage expression of IL-1beta and beta-cell expression of iNOS stimulated by the local release of IL-1 in rat islets. 15-d-delta 12,14-PGJ2 appears to be approximately 10-fold more effective at inhibiting resident islet macrophage activation (in response to TNF + LPS) than IL-1-induced nitrite production by beta-cells. Two mechanisms appear to be associated with the antiinflammatory actions of both 15-d-delta 12,14-PGJ2 and troglitazone: 1) the direct inhibition of cytokine- and endotoxin-stimulated iNOS and IL-1 transcription; and 2) the inhibition of IL-1 signaling, an event associated with PPAR-gamma agonist-induced activation of the heat shock response (as assayed by heat shock protein 70 expression). These findings indicate that the PPAR-gamma agonists, troglitazone and the J series of prostaglandins, are potent anti-inflammatory agents that prevent cytokine- and endotoxin-stimulated activation of peripheral and resident tissue macrophages and cytokine-induced iNOS expression by beta-cells by the inhibition of transcriptional activation and induction of the heat shock response.


Endogenous spatial attention during perceptual learning facilitates location transfer.
Covert attention and perceptual learning enhance perceptual performance. The relation between these two mechanisms is largely unknown. Previously, we showed that manipulating involuntary, exogenous spatial attention during training improved performance at trained and untrained locations, thus overcoming the typical location specificity. Notably, attention-induced transfer only occurred for high stimulus contrasts, at the upper asymptote of the psychometric function (i.e., via response gain). Here, we investigated whether and how voluntary, endogenous attention, the top-down and goal-based type of covert visual attention, influences perceptual learning. Twenty-six participants trained in an orientation discrimination task at two locations: half of participants received valid endogenous spatial precues (attention group), while the other half received neutral precues (neutral group). Before and after training, all participants were tested with neutral precues at two trained and two untrained locations. Within each session, stimulus contrast varied on a trial basis from very low (2%) to very high (64%). Performance was fit by a Weibull psychometric function separately for each day and location. Performance improved for both groups at the trained location, and unlike training with exogenous attention, at the threshold level (i.e., via contrast gain). The neutral group exhibited location specificity: Thresholds decreased at the trained locations, but not at the untrained locations. In contrast, participants in the attention group showed significant location transfer: Thresholds decreased to the same extent at both trained and untrained locations. These results indicate that, similar to exogenous spatial attention, endogenous spatial attention induces location transfer, but influences contrast gain instead of response gain.


Electroacupuncture Pretreatment Alleviates Cerebral Ischemic Injury Through α7 Nicotinic Acetylcholine Receptor-Mediated Phenotypic Conversion of Microglia.
Electroacupuncture (EA) pretreatment alleviates cerebral ischemic injury through α7 nicotinic acetylcholine receptor (α7nAChR). We attempted to investigate whether the phenotypic conversion of microglia was involved in the therapeutic effect of EA pretreatment in cerebral ischemia through α7nAChR. Adult male Sprague-Dawley (SD) rats were subjected to middle cerebral artery occlusion (MCAO) after EA or α7nAChR agonist N-(3R)-1-azabicyclo[2.2.2]oct-3-yl-furo[2,3-c]pyridine-5-carboxamide hydrochloride (PHA-543,613 hydrochloride) and antagonist α-bungarotoxin (α-BGT) pretreatment. Primary microglia were subjected to drug pretreatment and oxygen-glucose deprivation (OGD). The expressions of the classical activated phenotype (M1) microglia markers induced nitric oxide synthase (iNOS), interleukin-1β (IL-1β), and cluster of differentiation 86 (CD86); the alternative activated phenotype (M2) microglia markers arginase-1 (Arg-1), transforming growth factor-β1 (TGF-β1), and cluster of differentiation 206 (CD206); and the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and anti-inflammatory cytokines interleukin-4 (IL-4) and interleukin-10 (IL-10) in the ischemic penumbra or in the supernatant of primary microglia were analyzed. The infarction volume and neurological scores were assessed 72 h after reperfusion. The cell viability and lactate dehydrogenase (LDH) release of neurons co-cultured with microglia were analyzed using cell counting kit-8 (CCK-8) and LDH release assays. EA pretreatment decreased the expressions of M1 markers (iNOS, IL-1β, and CD86) and pro-inflammatory cytokines (TNF-α and IL-6), whereas it increased the expressions of M2 markers (Arg-1, TGF-β1, and CD206) and anti-inflammatory cytokines (IL-4 and IL-10) by activating α7nAChR. EA pretreatment also significantly reduced the infarction volume and improved the neurological deficit. The activation of α7nAChR in microglia relieved the inflammatory response of primary microglia subjected to OGD and attenuated the injury of neurons co-cultured with microglia. In conclusion, EA pretreatment alleviates cerebral ischemic injury through α7nAChR-mediated phenotypic conversion of microglia, which may be a new mechanism for the EA pretreatment-induced neuroprotection against cerebral ischemia.


Detection of viruses in young children with fever without an apparent source.
Fever without an apparent source is common in young children. Currently in the United States, serious bacterial infection is unusual. Our objective was to determine specific viruses that might be responsible. We enrolled children aged 2 to 36 months with temperature of 38°C or greater without an apparent source or with definite or probable bacterial infection being evaluated in the St Louis Children's Hospital Emergency Department and afebrile children having ambulatory surgery. Blood and nasopharyngeal swab samples were tested with an extensive battery of virus-specific polymerase chain reaction assays. One or more viruses were detected in 76% of 75 children with fever without an apparent source, 40% of 15 children with fever and a definite or probable bacterial infection, and 35% of 116 afebrile children (P < .001). Four viruses (adenovirus, human herpesvirus 6, enterovirus, and parechovirus) were predominant, being detected in 57% of children with fever without a source, 13% of children with fever and definite or probable bacterial infection, and 7% of afebrile children (P < .001). Thirty-four percent of 146 viral infections were detected only by polymerase chain reaction performed on blood. Fifty-one percent of children with viral infections and no evidence of bacterial infection were treated with antibiotics. Viral infections are frequent in children with fever without an apparent source. Testing of blood in addition to nasopharyngeal secretions expanded the range of viruses detected. Future studies should explore the utility of testing for the implicated viruses. Better recognition of viruses that cause undifferentiated fever in young children may help limit unnecessary antibiotic use.


Effects of neuromedin U-8 on stress responsiveness and hypothalamus-pituitary-adrenal axis activity in male C57BL/6J mice.
Neuromedin U (NMU) is a highly conserved neuropeptide that has been implicated in the stress response. To better understand how it influences various aspects of the stress response, we studied the effects of intracerebroventricular NMU-8 administration on stress-related behavior and activity of the hypothalamus-pituitary-adrenal (HPA) axis in male C57BL/6J mice. We investigated these NMU-8 effects when mice remained in their home cage and when they were challenged by exposure to forced swim stress. NMU-8 administration resulted in increased grooming behavior in mice that remained in their home cage and in a significant increase in c-Fos immunoreactivity in the paraventricular hypothalamus (PVH) and arcuate nucleus (ARC). Surprisingly, NMU-8 administration significantly decreased plasma corticosterone concentrations. Furthermore, NMU-8 administration increased immobility in the forced swim test in both naïve mice and mice that were previously exposed to swim stress. The effect of NMU-8 on c-Fos immunoreactivity in the PVH was dependent on previous exposure to swim stress given that we observed no significant changes in mice exposed for the first time to swim stress. In contrast, in the ARC we observed a significant increase in c-Fos immunoreactivity regardless of previous stress exposure. Interestingly, NMU-8 administration also significantly decreased plasma corticosterone concentrations in mice that were exposed to single forced swim stress, while this effect was no longer observed when mice were exposed to forced swim stress for a second time. Taken together, our data indicate that NMU-8 regulates stress responsiveness and suggests that its effects depend on previous stress exposure.


Interaction of spin-labeled HPMA-based nanoparticles with human blood plasma proteins - the introduction of protein-corona-free polymer nanomedicine.
In this paper, we revised the current understanding of the protein corona that is created on the surface of nanoparticles in blood plasma after an intravenous injection. We have focused on nanoparticles that have a proven therapeutic outcome. These nanoparticles are based on two types of biocompatible amphiphilic copolymers based on N-(2-hydroxypropyl)methacrylamide (HPMA): a block copolymer, poly(ε-caprolactone) (PCL)-b-poly(HPMA), and a statistical HPMA copolymer bearing cholesterol moieties, which have been tested both in vitro and in vivo. We studied the interaction of nanoparticles with blood plasma and selected blood plasma proteins by electron paramagnetic resonance (EPR), isothermal titration calorimetry, dynamic light scattering, and cryo-transmission electron microscopy. The copolymers were labeled with TEMPO radicals at the end of hydrophobic PCL or along the hydrophilic HPMA chains to monitor changes in polymer chain dynamics caused by protein adsorption. By EPR and other methods, we were able to probe specific interactions between nanoparticles and blood proteins, specifically low- and high-density lipoproteins, immunoglobulin G, human serum albumin (HSA), and human plasma. It was found that individual proteins and plasma have very low binding affinity to nanoparticles. We observed no hard corona around HPMA-based nanoparticles; with the exception of HSA the proteins showed no detectable binding to the nanoparticles. Our study confirms that a classical "hard corona-soft corona" paradigm is not valid for all types of nanoparticles and each system has a unique protein corona that is determined by the nature of the NP material.


Transfusion strategies for traumatic coagulopathy.
Uncontrolled bleeding is the most common preventable cause of death for patients with severe injury. Coagulopathy inevitably accompanies severe bleeding, exacerbated by the ongoing blood loss and the treatments administered. There is debate about the underlying pathophysiological mechanisms of early traumatic coagulopathy and uncertainty about whether injury induces a unique coagulopathy when compared to other forms of major haemorrhage. This review describes current understanding of the coagulopathy of major blood loss and focuses on the early coagulation changes that occur following severe injury. It then reports on the contemporary management of coagulopathic bleeding using new transfusion strategies. Finally this review presents some practical points to the delivery of transfusion for major blood loss in the modern hospital setting.


Internal optical forces in plasmonic nanostructures.
We present a computational study of the internal optical forces arising in plasmonic gap antennas, dolmen structures and split rings. We find that very strong internal forces perpendicular to the propagation direction appear in these systems. These internal forces show a rich behaviour with varying wavelength, incident polarisation and geometrical parameters, which we explain in terms of the polarisation charges induced on the structures. Various interesting and anomalous features arise such as lateral force reversal, optical pulling force, and circular polarisation-induced forces and torques along directions symmetry-forbidden for orthogonal linear polarisations. Understanding these effects and mastering internal forces in plasmonic nanostructures will be instrumental in implementing new functionalities in these nanophotonic systems.


Retinoic Acid signalling and the control of meiotic entry in the human fetal gonad.
The development of mammalian fetal germ cells along oogenic or spermatogenic fate trajectories is dictated by signals from the surrounding gonadal environment. Germ cells in the fetal testis enter mitotic arrest, whilst those in the fetal ovary undergo sex-specific entry into meiosis, the initiation of which is thought to be mediated by selective exposure of fetal ovarian germ cells to mesonephros-derived retinoic acid (RA). Aspects of this model are hard to reconcile with the spatiotemporal pattern of germ cell differentiation in the human fetal ovary, however. We have therefore examined the expression of components of the RA synthesis, metabolism and signalling pathways, and their downstream effectors and inhibitors in germ cells around the time of the initiation of meiosis in the human fetal gonad. Expression of the three RA-synthesising enzymes, ALDH1A1, 2 and 3 in the fetal ovary and testis was equal to or greater than that in the mesonephros at 8-9 weeks gestation, indicating an intrinsic capacity within the gonad to synthesise RA. Using immunohistochemistry to detect RA receptors RARα, β and RXRα, we find germ cells to be the predominant target of RA signalling in the fetal human ovary, but also reveal widespread receptor nuclear localization indicative of signalling in the testis, suggesting that human fetal testicular germ cells are not efficiently shielded from RA by the action of the RA-metabolising enzyme CYP26B1. Consistent with this, expression of CYP26B1 was greater in the human fetal ovary than testis, although the sexually-dimorphic expression patterns of the germ cell-intrinsic regulators of meiotic initiation, STRA8 and NANOS2, appear conserved. Finally, we demonstrate that RA induces a two-fold increase in STRA8 expression in cultures of human fetal testis, but is not sufficient to cause widespread meiosis-associated gene expression. Together, these data indicate that while local production of RA within the fetal ovary may be important in regulating the onset of meiosis in the human fetal ovary, mechanisms other than CYP26B1-mediated metabolism of RA may exist to inhibit the entry of germ cells into meiosis in the human fetal testis.


Effects of inspiratory flow waveforms on arterial blood gases and respiratory mechanics after open heart surgery.
The clinical usefulness of inspiratory flow pattern manipulation during mechanical ventilation remains unclear. The aim of this study was to investigate the effects of different inspiratory flow waveforms, i.e. constant, sinusoidal and decelerating, on arterial blood gases and respiratory mechanics, in mechanically ventilated patients. Eight patients recovering after open heart surgery for valvular replacement and/or coronary bypass were studied. The ventilator inspiratory flow waveform was changed according to a randomized sequence, keeping constant the other variables of the ventilator settings. We measured arterial blood gases, flow, volume and pressure at the proximal (airway opening pressure (Pao)) and distal (Ptr) ends of the endotracheal tubes before and after 30 min of mechanical ventilation with each inspiratory flow waveform. We computed breathing pattern, respiratory mechanics (pressures and dynamic elastance) and inspiratory work, which was then partitioned into its elastic and resistive components. We found that: 1) arterial oxygen tension (Pa,O2) and arterial carbon dioxide tension (Pa,CO2) were not affected by changes in the inspiratory flow waveform; and 2) peak Pao and Ptr were highest with sinusoidal inspiratory flow, whilst mean Pao and Ptr and total work of breathing were least with constant inspiratory flow, mainly because of a concomitant decrease in resistive work during constant flow inflation. The effects of the inspiratory flow profile on Pao, Ptr and total inspiratory work performed by the ventilator were mainly due to the resistive properties of the endotracheal tubes. We conclude that the ventilator inspiratory flow waveform can influence patients' respiratory mechanics, but has no impact on arterial oxygen and arterial carbon dioxide tension.


Mental health resilience in the adolescent offspring of parents with depression: a prospective longitudinal study.
Young people whose parents have depression have a greatly increased risk of developing a psychiatric disorder, but poor outcomes are not inevitable. Identification of the contributors to mental health resilience in young people at high familial risk is an internationally recognised priority. Our objectives were to identify protective factors that predict sustained good mental health in adolescents with a parent with depression and to test whether these contribute beyond what is explained by parent illness severity. The Early Prediction of Adolescent Depression study (EPAD) is a prospective longitudinal study of offspring of parents with recurrent depression. Parents with recurrent major depressive disorder, co-parents, and offspring (aged 9-17 years at baseline) were assessed three times over 4 years in a community setting. Offspring outcomes were operationalised as absence of mental health disorder, subthreshold symptoms, or suicidality on all three study occasions (sustained good mental health); and better than expected mental health (mood and behavioural symptoms at follow-up lower than predicted given severity of parental depression). Family, social, cognitive, and health behaviour predictor variables were assessed using interview and questionnaire measures. Between February and June, 2007, we screened 337 families at baseline, of which 331 were eligible. Of these, 262 completed the three assessments and were included in the data for sustained mental health. Adolescent mental health problems were common, but 53 (20%) of the 262 adolescents showed sustained good mental health. Index parent positive expressed emotion (odds ratio 1·91 [95% CI 1·31-2·79]; p=0·001), co-parent support (1·90 [1·38-2·62]; p<0·0001), good-quality social relationships (2·07 [1·35-3·18]; p=0·001), self-efficacy (1·49 [1·05-2·11]; p=0·03), and frequent exercise (2·96 [1·26-6·92]; p=0·01) were associated with sustained good mental health. Analyses accounting for parent depression severity were consistent, but frequent exercise only predicted better than expected mood-related mental health (β=-0·22; p=0·0004) not behavioural mental health, whereas index parents' expression of positive emotions predicted better than expected behavioural mental health (β=-0·16; p=0·01) not mood-related mental health. Multiple protective factors were required for offspring to be free of mental health problems (zero or one protective factor, 4% sustained good mental health; two protective factors, 10%; three protective factors, 13%, four protective factors, 38%; five protective factors, 48%). Adolescent mental health problems are common, but not inevitable, even when parental depression is severe and recurrent. These findings suggest that prevention programmes will need to enhance multiple protective factors across different domains of functioning. Sir Jules Thorn Charitable Trust, Economic and Social Research Council.


Crystal structure of nitrogen regulatory protein IIANtr from Neisseria meningitidis.
The NMB0736 gene of Neisseria meningitidis serogroup B strain MC58 encodes the putative nitrogen regulatory protein, IIANtr (abbreviated to NM-IIANtr). The homologous protein present in Escherichia coli is implicated in the control of nitrogen assimilation. As part of a structural proteomics approach to the study of pathogenic Neisseria spp., we have selected this protein for structure determination by X-ray crystallography. The NM-IIANtr was over-expressed in E. coli and was shown to be partially mono-phosphorylated, as assessed by mass spectrometry of the purified protein. Crystals of un-phosphorylated protein were obtained and diffraction data collected to 2.5 A resolution. The structure of NM-IIANtr was solved by molecular replacement using the coordinates of the E. coli nitrogen regulatory protein IIAntr [PDB: 1A6J] as the starting model. The overall fold of the Neisseria enzyme shows a high degree of similarity to the IIANtr from E. coli, and the position of the phosphoryl acceptor histidine residue (H67) is conserved. The orientation of an adjacent arginine residue (R69) suggests that it may also be involved in coordinating the phosphate group. Comparison of the structure with that of E. coli IIAmtl complexed with HPr [PDB: 1J6T] indicates that NM-IIANtr binds in a similar way to the HPr-like enzyme in Neisseria. The structure of NM-IIANtr confirms its assignment as a homologue of the IIANtr proteins found in a range of other Gram-negative bacteria. We conclude that the NM- IIANtr protein functions as part of a phosphorylation cascade which, in contrast to E. coli, shares the upstream phosphotransfer protein with the sugar uptake phosphoenolpyruvate:sugar phosphotransferase system (PTS), but in common with E. coli has a distinct downstream effector mechanism.


Evidence of limited contributions for intratumoral steroidogenesis in prostate cancer.
Androgen-deprivation therapy for prostate cancer (PC) eventually leads to castration-resistant PC (CRPC). Intratumoral androgen production might contribute to tumor progression despite suppressed serum androgen concentrations. In the present study, we investigated whether PC or CRPC tissue may be capable of intratumoral androgen synthesis. Steroidogenic enzyme mRNAs were quantified in hormonally manipulated human PC cell lines and xenografts as well as in human samples of normal prostate, locally confined and advanced PC, local nonmetastatic CRPC, and lymph node metastases. Overall, the majority of samples showed low or absent mRNA expression of steroidogenic enzymes required for de novo steroid synthesis. Simultaneous but low expression of the enzymes CYP17A1 and HSD3B1, essential for the synthesis of androgens from pregnenolone, could be detected in 19 of 88 patient samples. Of 19 CRPC tissues examined, only 5 samples expressed both enzymes. Enzymes that convert androstenedione to testosterone (AKR1C3) and testosterone to dihydrotestosterone (DHT; SRD5A1) were abundantly expressed. AKR1C3 expression was negatively regulated by androgens in the experimental models and was increased in CRPC samples. Expression of SRD5A1 was upregulated in locally advanced cancer, CRPC, and lymph node metastases. We concluded that intratumoral steroid biosynthesis contributes less than circulating adrenal androgens, implying that blocking androgen production and its intraprostatic conversion into DHT, such as via CYP17A1 inhibition, may represent favorable therapeutic options in patients with CRPC.


Use of social words in autobiographies and longevity.
To analyze the relationship between social word use in autobiographies and longevity. Although there is substantial evidence that our social relationships are associated with mortality, interpretation of this work is weakened by the limitations of assessing the social environment with structured questionnaires and interviews. By analyzing the word content of autobiographies, we could assess spontaneous indicators of important social relationships and relate them to longevity. This technique is less subject to social desirability reporting biases and more sensitive to aspects of the social environment that are central to how one experiences his or her social world. The autobiographies of 96 psychologists and 220 literary writers were digitized and scanned for social relationship word frequency via a computerized word counting program. Archival data were collected on birth and death dates, year of publication, place of birth, age when the autobiography was written, and sex. After controlling for sex, year of birth, and age at the time of writing, we found that higher use of words indicating social roles/integration (e.g., father, sister, neighbor, co-worker) was associated with an increased lifespan in both samples. Specific social categories assessing the use of family role terms (e.g., aunt, family, brother) and references to other individuals (e.g., they, we, us, everyone) also predicted longer life, but only in the sample of psychologists. Assessing social word use in autobiographies provided an indirect measure of social relationships that predicted longevity. This technique of analyzing writing samples may be useful in future archival research as well as in studies where it is desirable to study social relationships in an indirect fashion.


External injuries of Morelet's crocodile Crocodylus moreletii in Campeche, Mexico.
Analysis of external injuries in captive and free-ranging Morelet's crocodiles Crocodylus moreletii was performed in the northern wetlands of Campeche, Mexico. From March to September of 2007, a total of 52 free-ranging and 51 captive Morelet's crocodiles were studied. Captive crocodiles presented significantly more injuries. Sixteen free-ranging crocodiles presented some type of lesion, mostly superficial abrasions. Nineteen captive crocodiles presented lesions, mostly incisions from agonistic interactions. Overall, the injuries with highest prevalence were the incisions. The tail was the most frequently injured body region. Injuries were more common in adults than in other size classes. Conversely, the presence of lesions caused by the parasite Paratrichosoma spp. was greater in crocodiles captured in the coastal channels (mangrove habitat). The information presented here is important to understand some of the effects of individual interactions and to foresee and manage the consequences of conservation and management activities of crocodile populations.


Liquid detection with InGaAsP semiconductor lasers having multiple short external cavities.
A liquid detection system consisting of a diode laser with multiple short external cavities (MSXC's) is reported. The MSXC diode laser operates single mode on one of 18 distinct modes that span a range of 72 nm. We selected the modes by setting the length of one of the external cavities using a piezoelectric positioner. One can measure the transmission through cells by modulating the injection current at audio frequencies and using phase-sensitive detection to reject the ambient light and reduce 1/f noise. A method to determine regions of single-mode operation by the rms of the output of the laser is described. The transmission data were processed by multivariate calibration techniques, i.e., partial least squares and principal component regression. Water concentration in acetone was used to demonstrate the performance of the system. A correlation coefficient of R(2) = 0.997 and 0.29% root-mean-square error of prediction are found for water concentration over the range of 2-19%.


Arterial hypertension correlates with clinical outcome in colorectal cancer patients treated with first-line bevacizumab.
Arterial hypertension occurring during antiangiogenic therapy has been correlated with the biological inhibition of the vascular endothelial growth factor-related pathway and may represent a possible clinical marker for treatment efficacy. The aim of our study was to retrospectively assess if grades 2-3 hypertension were associated with response to bevacizumab, progression-free survival (PFS) and survival in metastatic colorectal cancer patients treated with first-line bevacizumab. Patients with histologically proven, metastatic colorectal cancer receiving bevacizumab as first-line therapy in combination with irinotecan and 5-fluorouracil were eligible for our analysis. Thirty-nine metastatic colorectal cancer patients were eligible. Eight patients (20%) developed grades 2-3 hypertension. A partial remission was observed in six of eight cases with bevacizumab-related hypertension (75%) and in 10 of 31 (32%) patients with no hypertension (P = 0.04). Median PFS was 14.5 months for patients showing bevacizumab-related hypertension, while it was 3.1 months in those without hypertension (P = 0.04). Median overall survival was not reached in patients with hypertension while it was 15.1 months in the remaining cases (P = 0.11). Our data indicate that bevacizumab-induced hypertension may represent an interesting prognostic factor for clinical outcome in advanced colorectal cancer patients receiving first-line bevacizumab.


Light scattering by a spheroidal particle.
The solution of electromagnetic scattering by a homogeneous prolate (or oblate) spheroidal particle with an arbitrary size and refractive index is obtained for any angle of incidence by solving Maxwell's equations under given boundary conditions. The method used is that of separating the vector wave equations in the spheroidal coordinates and expanding them in terms of the spheroidal wavefunctions. The unknown coefficients for the expansion are determined by a system of equations derived from the boundary conditions regarding the continuity of tangential components of the electric and magnetic vectors across the surface of the spheroid. The solutions both in the prolate and oblate spheroidal coordinate systems result in a same form, and the equations for the oblate spheroidal system can be obtained from those for the prolate one by replacing the prolate spheroidal wavefunctions with the oblate ones and vice versa. For an oblique incidence, the polarized incident wave is resolved into two components, the TM mode for which the magnetic vector vibrates perpendicularly to the incident plane and the TE mode for which the electric vector vibrates perpendicularly to this plane. For the incidence along the rotation axis the resultant equations are given in the form similar to the one for a sphere given by the Mie theory. The physical parameters involved are the following five quantities: the size parameter defined by the product of the semifocal distance of the spheroid and the propagation constant of the incident wave, the eccentricity, the refractive index of the spheroid relative to the surrounding medium, the incident angle between the direction of the incident wave and the rotation axis, and the angles that specify the direction of the scattered wave.


Differential microRNA-34a expression and tumor suppressor function in retinoblastoma cells.
The role of miR-34a, a p53-regulated microRNA, in retinoblastoma (RB) was investigated. The expression of miR-34 family members in RB cells was determined by semiquantitative RT-PCR and real-time qPCR. Regulation of miR-34a expression by p53-activating compounds was determined by qPCR analysis. The tumor suppressor functions of miR-34a in RB cell lines were determined by tetrazolium-based cell growth assay and by caspase-3/7 and activated caspase-3 apoptotic activity assays. Additive growth inhibitory properties of miR-34a in combination with topotecan were determined by cell growth assay. miR-34a targets in RB cells were identified by real-time qPCR expression analysis of previously reported and GenMiR++-predicted mRNAs. Differential miR-34a and miR-34b expression was observed in RB cell lines and tumor samples. miR-34a expression could be increased in Y79 cells, but not Weri-Rb1 cells, after p53 activation. This differential regulation was not caused by genomic alterations at the miR-34a p53 binding site or mature gene. Exogenous miR-34a inhibited Y79 and Weri-Rb1 cell growth and increased apoptotic activity in Y79 cells. Increased inhibition of Y79 and Weri-Rb1 cell growth was observed with combination miR-34a and topotecan treatment. mRNA expression changes were observed in 7 of 7 previously reported and 13 of 18 GenMiR++-predicted miR-34a targets after transfection of Y79 cells with miR-34a compared with negative control microRNA. miR-34a functions as a tumor suppressor in RB cells and is a potential therapeutic target. Differential expression, regulation, and activity of miR-34a in RB cells may suggest further p53 pathway inactivation in RB.


Hierarchical multi-grain models improve descriptions of species' environmental associations, distribution, and abundance.
The characterization of species' environmental niches and spatial distribution predictions based on them are now central to much of ecology and conservation, but implicitly requires decisions about the appropriate spatial scale (i.e., grain) of analysis. Ecological theory and empirical evidence suggest that range-resident species respond to their environment at two characteristic, hierarchical spatial grains: (1) response grain, the (relatively fine) grain at which an individual uses environmental resources, and (2) occupancy grain, the (relatively coarse) grain equivalent to a typical home range. We use a multi-grain (MG) occupancy model, aided by fine-grain remotely sensed imagery, to simultaneously estimate species-environment associations at both grains, conduct grain optimization to measure response grain, and apply this analysis framework to an example species: a medium-sized bird (Tockus deckeni) in a heterogeneous East African landscape. Based on home range analysis of movement data, we calculate an occupancy grain of 1 km for T. deckeni. Using a grain optimization procedure across 32 grains from 10 to 500 m, we identify 60 m as the most strongly supported response grain for a suite of environmental variables, slightly coarser than opportunistic behavioral observations would have suggested. Validation confirms that the accuracy of the optimized MG occupancy model substantially exceeds that of equivalent single-grain (SG) occupancy models. We further use a simulation approach to assess the potential impacts of accounting for the multi-scale structure of species' environmental requirements on estimates of population size. We find that the more strongly supported MG approach consistently predicts a minimum population size in the study landscape that is much lower than that provided by the SG model. This suggests that SG approaches commonly used in conservation applications could lead to overly optimistic abundance and population estimates, and that the MG approach may be more appropriate for supporting species conservation goals. More generally, we conclude that multi-grain approaches of the sort presented, and increasingly enabled by growing high-resolution remotely sensed data, hold great promise for offering a more mechanistic framework for assessing the appropriate grain(s) for population monitoring and management and enable more reliable estimates of abundances and species' distributions.


Examining the role of social support and group cohesion in exercise compliance.
The present study sought to examine the role of social support and group cohesion in the compliance behavior of females required to exercise for health-related reasons. Compliance behavior was assessed as attendance and dropout behavior. Participants (N = 49) drawn from a clinical exercise group completed questionnaires to assess cohesion and social support. Discriminant function analyses were able to discriminate between high and low attenders as well as between dropouts and graduates. Participants reporting high scores on the support provision of reliable alliance and the cohesion factor of ATG-Task, along with low scores on the social support provision of guidance, were more likely to attend a higher proportion of classes. For dropouts, participants with higher scores on the support provision of reliable alliance were less likely to drop out. As a secondary objective, it was found that cohesion and social support variables both contributed to the successful prediction of attendance.


A Pilot Study of Mindfulness-Based Exposure Therapy in OEF/OIF Combat Veterans with PTSD: Altered Medial Frontal Cortex and Amygdala Responses in Social-Emotional Processing.
Combat-related posttraumatic stress disorder (PTSD) is common among returning veterans, and is a serious and debilitating disorder. While highly effective treatments involving trauma exposure exist, difficulties with engagement and early drop may lead to sub-optimal outcomes. Mindfulness training may provide a method for increasing emotional regulation skills that may improve engagement in trauma-focused therapy. Here, we examine potential neural correlates of mindfulness training and in vivo exposure (non-trauma focused) using a novel group therapy [mindfulness-based exposure therapy (MBET)] in Afghanistan (OEF) or Iraq (OIF) combat veterans with PTSD. OEF/OIF combat veterans with PTSD (N = 23) were treated with MBET (N = 14) or a comparison group therapy [Present-centered group therapy (PCGT), N = 9]. PTSD symptoms were assessed at pre- and post-therapy with Clinician Administered PTSD scale. Functional neuroimaging (3-T fMRI) before and after therapy examined responses to emotional faces (angry, fearful, and neutral faces). Patients treated with MBET had reduced PTSD symptoms (effect size d = 0.92) but effect was not significantly different from PCGT (d = 0.43). Improvement in PTSD symptoms from pre- to post-treatment in both treatment groups was correlated with increased activity in rostral anterior cingulate cortex, dorsal medial prefrontal cortex (mPFC), and left amygdala. The MBET group showed greater increases in amygdala and fusiform gyrus responses to Angry faces, as well as increased response in left mPFC to Fearful faces. These preliminary findings provide intriguing evidence that MBET group therapy for PTSD may lead to changes in neural processing of social-emotional threat related to symptom reduction.


Biotin administration improves the impaired glucose tolerance of streptozotocin-induced diabetic Wistar rats.
The effect of biotin administration on the glucose tolerance of streptozotocin (STZ)-induced diabetic Wistar rats was investigated. STZ-induced diabetes was induced by intraperitoneal injection of streptozotocin (45 mg/kg body weight as a single dose). The impaired glucose tolerance in response to an oral glucose load (1.8g per kg body weight) in STZ-induced diabetic rats (STZ-rat) was partially improved by intraperitoneal administration of biotin for 15 days (100 micrograms/rat/day). However, a recovery in the STZ-rat's insulin secretion was not found after biotin administration. To help clarify the mechanism underlying the improvement in glucose tolerance seen with biotin treatment, glucokinase and hexokinase activities were determined in the liver and pancreas. In STZ-rats that had received biotin (STZ-biotin rats), glucokinase activity was higher by 3.4-fold in liver and by 2.4-fold in pancreas than in the STZ-rats. The biotin level of STZ-rats was significantly lower in the liver and pancreas than that of the control rats (no STZ administration); but in STZ-biotin rats, the level in these organs recovered to the control level. These results demonstrate that injected biotin can improve glucose handling without increasing insulin secretion in STZ-rats.


Establishment of haematological and immunological reference values for healthy Tanzanian children in Kilimanjaro Region.
To determine the normal haematological and immunological reference intervals for healthy Tanzanian children. We analysed data from 655 HIV-seronegative, healthy children from 1 month to 18 years of age from the Kilimanjaro Region of Tanzania for this cross-sectional study. Median and 95% reference ranges were determined for haematological and immunological parameters and analysed by age cohorts, and by gender for adolescents. Median haemoglobin (Hb) and haematocrit (Hct) for all age groups were higher than established East African reference intervals. Compared to U.S. intervals, reference ranges encompassed lower values for Hb, Hct, mean corpuscular volume, and platelets. Applying the U.S. National Institute of Health Division of AIDS (DAIDS) adverse event grading criteria commonly used in clinical trials to the reference range participants, 128 (21%) of 619 children would be classified as having an adverse event related to Hb level. CD4-positive T-lymphocyte absolute counts declined significantly with increasing age (P < 0.0001). For those aged under five years, CD4-positive T-lymphocyte percentages are lower than established developed country medians. Country-specific reference ranges are needed for defining normal laboratory parameters among children in Africa. Knowledge of appropriate reference intervals is critical not only for providing optimal clinical care, but also for enrolling children in medical research. Knowledge of normal CD4-positive T-lymphocyte parameters in this population is especially important for guiding the practice of HIV medicine in Tanzania.


Sleep Disturbance and the Risk of Cognitive Decline or Clinical Conversion in the ADNI Cohort.
We investigated the relationship between sleep disturbance and cognitive decline or clinical conversion in individuals with normal cognition (CN), as well as those with mild cognitive impairment (MCI) and dementia due to Alzheimer disease (AD-dementia). Secondary analysis of 1,629 adults between 48 and 91 years of age with up to 24 months of follow-up from the ADNI (Alzheimer's Disease Neuroimaging Initiative), a longitudinal cohort study. Sleep disturbance was not associated with decline in memory, executive function, or global cognition. The presence of sleep disturbance did not significantly increase the risk of diagnostic conversion in CN, early MCI, or late MCI participants. This study investigated the effect of sleep disturbance on cognitive decline using several outcomes and does not support the hypothesis that sleep disturbance predicts subsequent cognitive decline.


Comparing quantitative and qualitative indices of the donated corneas maintained in Optisol-GS with those kept in Eusol-C.
This study was conducted to compare the quantitative and qualitative indices of the donated corneas maintained in either Optisol-GS or Eusol-C storage media. In an ante-grade single blind study, two corneas from each donor with a death to preservation time of less than 30 h and with a minimum of "an apparent good cornea rating" were maintained in corneal storage media; randomly one in Optisol-GS and the other in Eusol-C. Slit-lamp biomicroscopic and specular microscopic examinations were performed on days 1 and 7. The results of the qualitative and quantitative indices and the final cornea rating were recorded. Statistical analyses were performed to evaluate any differences between the two media. 180 corneas from 90 donors with an age range of 29.3 ± 22.4 years were allocated in two groups: 90 corneas in Optisol-GS and the other 90 in Eusol-C. Five corneas in Optisol-GS and four corneas in Eusol-C were excluded from the study due to lack of specular images. There was no significant change of endothelial rating from day 1 to day 7 between two media (P = 0.175). As the maintenance time of the donated corneas increased, no significant difference was noted between the two groups in terms of endothelial indices, stromal edema, and Descemet's folding. This study shows no superiority between Eusol-C and Optisol-GS in short term preservation of donated corneas.


Biocrust morphogroups provide an effective and rapid assessment tool for drylands.
Biological soil crusts (biocrusts) occur across most of the world's drylands and are sensitive indicators of dryland degradation. Accounting for shifts in biocrust composition is important for quantifying integrity of arid and semi-arid ecosystems, but the best methods for assessing biocrusts are uncertain. We investigate the utility of surveying biocrust morphogroups, a reduced set of biotic classes, compared to species data, for detecting shifts in biocrust composition and making inference about dryland degradation.We used multivariate regression tree (MRT) analyses to model morphogroup abundance, species abundance and species occurrence data from two independent studies in semi-arid open woodlands of south-eastern Australia. We advanced the MRT method with a 'best subsets' model selection procedure, which improved model stability and prediction.Biocrust morphogroup composition responded strongly to surrogate variables of ecological degradation. Further, MRT models of morphogroup data had stronger explanatory power and predictive power than MRT models of species abundance or occurrence data. We also identified morphogroup indicators of degraded and less degraded sites in our study region.Synthesis and applications. Sustainable management of drylands requires methods to assess shifts in ecological integrity. We suggest that biocrust morphogroups are highly suitable for assessment of dryland integrity because they allow for non-expert, rapid survey and are informative about ecological function. Furthermore, morphogroups were more robust than biocrust species data, showed a strong response to ecological degradation and were less influenced by environmental variation, and models of morphogroup abundance were more predictive.


Effects of electro-acupuncture on anovulation in women with polycystic ovary syndrome.
The present study was designed to evaluate if electro-acupuncture (EA) could affect oligo-/anovulation and related endocrine and neuroendocrine parameters in women with polycystic ovary syndrome (PCOS). Twenty-four women (between the ages of 24 and 40 years) with PCOS and oligo-/amenorrhea were included in this non-randomized, longitudinal, prospective study. The study period was defined as the period extending from 3 months before the first EA treatment, to 3 months after the last EA treatment (10-14 treatments), in total 8-9 months. The menstrual and ovulation patterns were confirmed by recording of vaginal bleedings and by daily registrations of the basal body temperature (BBT). Blood samples were collected within a week before the first EA, within a week after the last EA and 3 months after EA. Nine women (38%) experienced a good effect. They displayed a mean of 0.66 ovulations/woman and month in the period during and after the EA period compared to a mean of 0.15 before the EA period (p=0.004). Before EA, women with a good effect had a significantly lower body-mass index (BMI) (p<0.001), waist-to-hip circumference ratio (WHR) (p=0.0058), serum testosterone concentration (p=0.0098), serum testosterone/sex hormone binding globulin (SHBG) ratio (p=0.011) and serum basal insulin concentration (p=0.0054), and a significantly higher concentration of serum SHBG (p=0.040) than did those women with no effect. Repeated EA treatments induce regular ovulations in more than one third of the women with PCOS. The group of women with good effect had a less androgenic hormonal profile before treatment and a less pronounced metabolic disturbance compared with the group with no effect. For this selected group EA offers an alternative to pharmacological ovulation induction.


Farmers' use and adaptation of improved climbing bean production practices in the highlands of Uganda.
Climbing beans offer potential for sustainable intensification of agriculture, but their cultivation constitutes a relatively complex technology consisting of multiple components or practices. We studied uptake of improved climbing bean production practices (improved variety, input use and management practices) through co-designed demonstrations and farmer-managed adaptation trials with 374 smallholder farmers in eastern and southwestern Uganda. A sub-set of these farmers was monitored one to three seasons after introduction. About 70% of the farmers re-planted climbing beans one season after the adaptation trial, with significant differences between eastern (50%) and southwestern Uganda (80-90%). Only 1% of the farmers used all of the improved practices and 99% adapted the technology. On average, farmers used half of the practices in different combinations, and all farmers used at least one of the practices. Yield variability of the trials was large and on average, trial plots did not yield more than farmers' own climbing bean plots. Yet, achieved yields did not influence whether farmers continued to cultivate climbing bean in the subsequent season. Uptake of climbing beans varied with household characteristics: poorer farmers cultivated climbing beans more often but used fewer of the best-bet practices; male farmers generally used more practices than female farmers. Planting by poorer farmers resulted in adaptations such as growing climbing beans without fertilizer and with fewer and shorter stakes. Other relationships were often inconsistent and farmers changed practices from season to season. The diversity of farmer responses complicates the development of recommendation domains and warrants the development of a basket of options from which farmers can choose. Our study shows how adoption of technologies consisting of multiple components is a complicated process that is hard to capture through the measurement of an adoption rate at a single point in time.


Anti-mullerian hormone as a predictor of ovarian reserve and ovarian response in IVF women from Gaza strip.
Back ground: Careful evaluation of patients and proper treatment with right techniques are essential for successful outcome of assisted reproduction. To obtain satisfactory results, it is necessary to assess ovarian reserve before planning treatment. To evaluate anti-mullerian hormone as a predictor of fertility potential in terms of ovarian reserve and ovarian response reflected by antral follicles and mature oocyte counts in response to menotrophin stimulation in in vitro fertilization (IVF) women from Gaza Strip. This prospective cohort study consisted of 81 women (mean age 28.7 years) attending IVF at Al-Basma Fertility Center in Gaza City. Blood withdrawal for antimullerian hormone measurement was performed in all the patients and the number of oocytes and embryos were recorded. The total number of retrieved oocytes was inversely associated with age (12.5±4.5, 11.0±5.4 and 6.9±4.7 at age ≤25, 26-35 and >35 years, respectively (F=4.793 and p=0.011). The ovarian response to Menotrophin (FSH 75IU, LH 75 IU) stimulation was better for younger age. There was a significant positive association between ovarian response in terms of total number of oocytes and antimullerian hormone levels. The maximum level of antimullerian hormone was observed in females who achieved positive pregnancies (4.5±2.5 ng/mL) followed by negative pregnancies (2.9±1.8 ng/mL) with significant differences (F=6.862 and p=0.002). Correlation coefficient revealed that the number of mature oocytes showed strong positive correlation with the antimullerian hormone levels (r=0.469, p=0.001). Anti-mullerian hormone can be used in IVF programs as a good predictor of ovarian reserve and ovarian response.


Effects of erythropoietin on erythroid colony formation in uremic rabbit bone marrow cultures.
Erythropoietin-responsive stem cell (ERC) kinetics in anephric uremic rabbits were studied in vitro using the growth of erythroid colonies in a methyl cellulose system in cultures with and without the addition of erythropoietin (ESF). Approximately 68 hr after bilateral nephrectomy, an increase in BUN and decreases in hematocrit and marrow erythroid cellularity were seen. However, the numbers of erythroid colonies formed in response to ESF on plates inoculated with 2 times 10-5 cells were greater in anephric rabbit marrows than in normal controls. In addition, the numbers of erythroid colonies produced by the uremic and normal marrows in the presence of ESF were increased in proportion to the number of precursors plated. These findings suggest that, in uremia, the concentration of ERC is increased and that the ERC are capable of responding normally to ESF. The increase in the number of erythroid colonies of uremia may be due to the undisturbed flow of uncommitted hematopoietic stem cells into the ERC compartment in the presence of a delay of differentiation of ERC into heme-synthesizing nucleated erythroid cells due to a lack of ESF.


Surveillance for traumatic brain injury deaths--United States, 1989-1998.
Data indicate that approximately 50,000 U.S. residents die as a result of traumatic brain injury (TBI) annually. Survivors of TBI are often left with neuropsychologic impairments that result in disabilities affecting work or social activity. During 1979-1992, TBI-related death rates declined 22%, from 24.6 to 19.3 deaths/100,000 population. This report describes the epidemiology and trends in TBI-related mortality during 1989-1998. January 1, 1989-December 31, 1998. The National Center for Health Statistics (NCHS) Multiple Cause of Death public use data were analyzed for this study. During 1989-1998, an annual average of 53,288 deaths (range: 51,848-54,501) among U.S. residents were associated with TBI. TBI-related death rates declined 11.4%, from 21.9 to 19.4/100,000 population. The major causes of TBI-related deaths were firearm-related (40%), motor-vehicle-related (34%), and fall-related (10%). The leading causes of TBI-related deaths differed among age groups. Among youths aged 0-19 years, motor-vehicle-related TBIs were the leading cause; among persons aged 20-74 years, firearm-related TBIs were the leading cause; and among persons aged > or = 75 years, fall-related TBIs were the leading cause. Comparing rates in 1989 with rates in 1998, motor-vehicle-related causes declined by 22%; the majority of this decline occurred during the first 5 years of the period. During 1989-1998, firearm-related TBI-related deaths declined by 14%; approximately all of this decline occurred during the last 5 years of the period. In contrast, fall-related TBI-related death rates increased by 25% during the period. This analysis of mortality data identifies recent trends in TBI-related deaths occurring during 1989-1998. Fall-related TBI death rates have increased throughout the period. Firearm-related TBI death rates, which were increasing in the early 1990s, declined. Motor-vehicle-related TBI death rates, which were decreasing until the mid-1990s, have since demonstrated only a limited change. More current population-based epidemiologic studies of TBI are needed to assess recent trends of etiologic factors, provide additional guidance for public policy, and evaluate prevention strategies. Despite the decline in fatal TBI incidence, TBI morbidity and mortality remains a public health challenge. Public health, law enforcement, and transportation safety professionals can address these challenges by implementing effective interventions based on a thorough assessment of the factors that influence health-related behaviors.


Inhibitory phosphorylation of the APC regulator Hct1 is controlled by the kinase Cdc28 and the phosphatase Cdc14.
Exit from mitosis requires inactivation of mitotic cyclin-dependent kinases (CDKs). A key mechanism of CDK inactivation is ubiquitin-mediated cyclin proteolysis, which is triggered by the late mitotic activation of a ubiquitin ligase known as the anaphase-promoting complex (APC). Activation of the APC requires its association with substoichiometric activating subunits termed Cdc20 and Hct1 (also known as Cdh1). Here, we explore the molecular function and regulation of the APC regulatory subunit Hct1 in Saccharomyces cerevisiae. Recombinant Hct1 activated the cyclin-ubiquitin ligase activity of APC isolated from multiple cell cycle stages. APC isolated from cells arrested in G1, or in late mitosis due to the cdc14-1 mutation, was more responsive to Hct1 than APC isolated from other stages. We found that Hct1 was phosphorylated in vivo at multiple CDK consensus sites during cell cycle stages when activity of the cyclin-dependent kinase Cdc28 is high and APC activity is low. Purified Hct1 was phosphorylated in vitro at these sites by purified Cdc28-cyclin complexes, and phosphorylation abolished the ability of Hct1 to activate the APC in vitro. The phosphatase Cdc14, which is known to be required for APC activation in vivo, was able to reverse the effects of Cdc28 by catalyzing Hct1 dephosphorylation and activation. We conclude that Hct1 phosphorylation is a key regulatory mechanism in the control of cyclin destruction. Phosphorylation of Hct1 provides a mechanism by which Cdc28 blocks its own inactivation during S phase and early mitosis. Following anaphase, dephosphorylation of Hct1 by Cdc14 may help initiate cyclin destruction.


Provider cultural competency, client satisfaction, and engagement in home-based programs to treat child abuse and neglect.
Home-based programs to treat child abuse and neglect suffer from high rates of attrition, limiting their impact. Thus, research is needed to identify factors related to client engagement. Using data (N = 1,305) from a statewide family preservation program, this study investigated the role of program type (i.e., SafeCare(®) [SC] vs. Services as Usual [SAU]) and client perceived provider cultural competence on client satisfaction and engagement with services. Families in SC completed more treatment goals than those in SAU. In addition, provider cultural competence and client satisfaction were higher in SC than in SAU. Higher provider cultural competence was associated with higher goal attainment and satisfaction, and these effects partially mediated the service program differences. The effects of service type and cultural competence on goal attainment and satisfaction varied somewhat by client ethnicity. Findings suggest that clients receiving manualized programs for child maltreatment may be more likely to meet their goals and may perceive such programs to be culturally appropriate and satisfactory.


Enhanced inertial focusing of microparticles and cells by integrating trapezoidal microchambers in spiral microfluidic channels.
In this work, manipulating width and equilibrium position of fluorescent microparticles in spiral microchannel fractionation devices by embedding microchambers along the last turn of a spiral is reported. Microchambers with different shapes and sizes were tested at Reynolds numbers between 15.7 and 156.6 (100-1000 μL min-1) to observe focusing of 2, 5 and 10 μm fluorescent microparticles. This paper also discusses the fabrication process of the microfluidic chips with femtosecond laser ablation on glass wafers, as well as a particle imaging velocimetry (μPIV) study of microparticle trajectories inside a microchamber. It could be demonstrated with an improved final design with inclined microchamber side walls, that the 2 μm particle equilibrium position is shifted towards the inner wall by ∼27 μm and the focusing line's width is reduced by ∼18 μm. Finally, Saccharomyces cerevisiae yeast cells were tested in the final chip and a cell focusing efficiency of 99.1% is achieved.


The Mortality and Myocardial Effects of Antidepressants Are Moderated by Preexisting Cardiovascular Disease: A Meta-Analysis.
Antidepressants (ADs) are commonly prescribed medications, but their long-term health effects are debated. ADs disrupt multiple adaptive processes regulated by evolutionarily ancient biochemicals, potentially increasing mortality. However, many ADs also have anticlotting properties that can be efficacious in treating cardiovascular disease. We conducted a meta-analysis assessing the effects of ADs on all-cause mortality and cardiovascular events in general-population and cardiovascular-patient samples. Two reviewers independently assessed articles from PubMed, EMBASE, and Google Scholar for AD-related mortality controlling for depression and other comorbidities. From these articles, we extracted information about cardiovascular events, cardiovascular risk status, and AD class. We conducted mixed-effect meta-analyses testing sample type and AD class as moderators of all-cause mortality and new cardiovascular events. Seventeen studies met our search criteria. Sample type consistently moderated health risks. In general-population samples, AD use increased the risks of mortality (HR = 1.33, 95% CI: 1.14-1.55) and new cardiovascular events (HR = 1.14, 95% CI: 1.08-1.21). In cardiovascular patients, AD use did not significantly affect risks. AD class also moderated mortality, but the serotonin reuptake inhibitors were not significantly different from tricyclic ADs (TCAs) (HR = 1.10, 95% CI: 0.93-1.31, p = 0.27). Only "other ADs" were differentiable from TCAs (HR = 1.35, 95% CI: 1.08-1.69). Mortality risk estimates increased when we analyzed the subset of studies controlling for premedication depression, suggesting the absence of confounding by indication. The results support the hypothesis that ADs are harmful in the general population but less harmful in cardiovascular patients.


The past matters: estimating intrinsic hookworm transmission intensity in areas with past mass drug administration to control lymphatic filariasis.
Current WHO guidelines for soil-transmitted helminth (STH) control focus on mass drug administration (MDA) targeting preschool-aged (pre-SAC) and school-aged children (SAC), with the goal of eliminating STH as a public health problem amongst children. Recently, attention and funding has turned towards the question whether MDA alone can result in the interruption of transmission for STH. The lymphatic filariasis (LF) elimination programme, have been successful in reaching whole communities. There is the possibility of building upon the infrastructure created for these LF-programmes to enhance the control of STH. Using hookworm as an example, we explore what further MDA coverage might be required to induce interruption of transmission for hookworm in the wake of a successful LF programme. Analyses based on the model of STH transmission and MDA impact predict the effects of previous LF control by MDA over five years, on a defined baseline prevalence of STH in an area with a defined transmission intensity (the basic reproductive number R0). If the LF MDA programme achieved a high coverage (70, 70 and 60% for pre-SAC, SAC and adults, respectively) we expect that in communities with a hookworm prevalence of 15%, after 5 years of LF control, the intrinsic R0 value in that setting is 2.47. By contrast, if lower LF coverages were achieved (40, 40 and 30% for pre-SAC, SAC and adults, respectively), with the same prevalence of 15% at baseline (after 5 years of LF MDA), the intrinsic hookworm R0 value is predicted to be 1.67. The intrinsic R0 value has a large effect on the expected successes of follow-up STH programmes post LF MDA. Consequently, the outcomes of identical programmes may differ between these communities. To design the optimal MDA intervention to eliminate STH infections, it is vital to have information on historical MDA programmes and baseline prevalence to estimate the intrinsic transmission intensity for the defined setting (R0). The baseline prevalence alone is not sufficient to inform policy for the control of STH, post cessation of LF MDA, since this will be highly dependent on the intensity and effectiveness of past programmes and the intrinsic transmission intensity of the dominant STH species in any given setting.


Restrictive versus liberal blood transfusion for acute upper gastrointestinal bleeding (TRIGGER): a pragmatic, open-label, cluster randomised feasibility trial.
Transfusion thresholds for acute upper gastrointestinal bleeding are controversial. So far, only three small, underpowered studies and one single-centre trial have been done. Findings from the single-centre trial showed reduced mortality with restrictive red blood cell (RBC) transfusion. We aimed to assess whether a multicentre, cluster randomised trial is a feasible method to substantiate or refute this finding. In this pragmatic, open-label, cluster randomised feasibility trial, done in six university hospitals in the UK, we enrolled all patients aged 18 years or older with new presentations of acute upper gastrointestinal bleeding, irrespective of comorbidity, except for exsanguinating haemorrhage. We randomly assigned hospitals (1:1) with a computer-generated randomisation sequence (random permuted block size of 6, without stratification or matching) to either a restrictive (transfusion when haemoglobin concentration fell below 80 g/L) or liberal (transfusion when haemoglobin concentration fell below 100 g/L) RBC transfusion policy. Neither patients nor investigators were masked to treatment allocation. Feasibility outcomes were recruitment rate, protocol adherence, haemoglobin concentration, RBC exposure, selection bias, and information to guide design and economic evaluation of the phase 3 trial. Main exploratory clinical outcomes were further bleeding and mortality at day 28. We did analyses on all enrolled patients for whom an outcome was available. This trial is registered, ISRCTN85757829 and NCT02105532. Between Sept 3, 2012, and March 1, 2013, we enrolled 936 patients across six hospitals (403 patients in three hospitals with a restrictive policy and 533 patients in three hospitals with a liberal policy). Recruitment rate was significantly higher for the liberal than for the restrictive policy (62% vs 55%; p=0·04). Despite some baseline imbalances, Rockall and Blatchford risk scores were identical between policies. Protocol adherence was 96% (SD 10) in the restrictive policy vs 83% (25) in the liberal policy (difference 14%; 95% CI 7-21; p=0·005). Mean last recorded haemoglobin concentration was 116 (SD 24) g/L for patients on the restrictive policy and 118 (20) g/L for those on the liberal policy (difference -2·0 [95% CI -12·0 to 7·0]; p=0·50). Fewer patients received RBCs on the restrictive policy than on the liberal policy (restrictive policy 133 [33%] vs liberal policy 247 [46%]; difference -12% [95% CI -35 to 11]; p=0·23), with fewer RBC units transfused (mean 1·2 [SD 2·1] vs 1·9 [2·8]; difference -0·7 [-1·6 to 0·3]; p=0·12), although these differences were not significant. We noted no significant difference in clinical outcomes. A cluster randomised design led to rapid recruitment, high protocol adherence, separation in degree of anaemia between groups, and non-significant reduction in RBC transfusion in the restrictive policy. A large cluster randomised trial to assess the effectiveness of transfusion strategies for acute upper gastrointestinal bleeding is both feasible and essential before clinical practice guidelines change to recommend restrictive transfusion for all patients with acute upper gastrointestinal bleeding. NHS Blood and Transplant Research and Development.


[How to define the species barrier to pathogen transmission?].
A given microbial pathogen usually targets a restricted number of animal species. Some pathogens can be transmitted to humans from another animal species, either directly (rabies, brucellosis, etc.) or through a vector (Lyme's disease, West Nile fever, etc.). Few infectious agents with animal reservoirs infect humans, and even fewer are capable of human-human transmission. This is attributed to the "species barrier", a simplistic concept that in fact involves a series of conditions for successful inter-species transmission. These include access to an infectable surface, multiplication on that surface, colonisation, invasion, multiplication inside the new host, and resistance to innate and adaptive immune mechanisms. Each of these steps requires a specific ligand-receptor interaction. The full series of events must be "reprogrammed" for efficient implantation in a new host. These changes occur through mutations or genetic exchanges. Direct human-to-human transmission often requires additional adaptive modifications.


Brucella species survey in polar bears (ursus maritimus) of northern Alaska.
We report on the presence of specific antibodies to Brucella spp. and Yersinia enterocolitica in polar bears (Ursus maritimus) from northern Alaska (southern Beaufort Sea) during 2003-2006. Based on numerous known stressors (e.g., climate change and loss of sea ice habitat, contaminants), there is increased concern regarding the status of polar bears. Considering these changes, it is important to assess exposure to potentially pathogenic organisms and to improve understanding of transmission pathways. Brucella or specific antibodies to Brucella spp. has been reported in marine mammals. Various assays were used to elucidate the pathway or source of exposure (e.g., "marine" vs. "terrestrial" Brucella spp.) of northern Alaska polar bears to Brucella spp. The standard plate test (SPT) and the buffered Brucella antigen card test (BBA) were used for initial screening for antibodies specific to Brucella. We then evaluated positive reactors (presence of serum antibody specific for Brucella spp.) using immunoblots and competitive enzyme-linked immunosorbent assay (cELISA; based on pinniped-derived Brucella spp. antigen). Annual prevalence of antibody (BBA and SPT) for Brucella spp. ranged from 6.8% to 18.5% over 2003-2006, with an overall prevalence of 10.2%. Prevalence of Brucella spp. antibody did vary by age class. Western blot analyses indicated 17 samples were positive for Brucella spp. antibody; of these, 13 were negative by marine (pinniped) derived Brucella antigen cELISA and four were positive by marine cELISA. Of the four samples positive for Brucella antibody by marine cELISA, three cross-reacted with Y. enterocolitica and Brucella spp. (one sample was Brucella negative and Y. enterocolitica positive). It appears the polar bear antibody does not react with the antigens used on the marine cELISA assay, potentially indicating a terrestrial (nonpinniped) source of Brucella spp.


Estrogen regulates hepcidin expression via GPR30-BMP6-dependent signaling in hepatocytes.
Hepcidin, a liver-derived iron regulatory protein, plays a crucial role in iron metabolism. It is known that gender differences exist with respect to iron storage in the body; however, the effects of sex steroid hormones on iron metabolism are not completely understood. We focused on the effects of the female sex hormone estrogen on hepcidin expression. First, ovariectomized (OVX) and sham-operated mice were employed to investigate the effects of estrogen on hepcidin expression in an in vivo study. Hepcidin expression was decreased in the livers of OVX mice compared to the sham-operated mice. In OVX mice, bone morphologic protein-6 (BMP6), a regulator of hepcidin, was also found to be downregulated in the liver, whereas ferroportin (FPN), an iron export protein, was upregulated in the duodenum. Both serum and liver iron concentrations were elevated in OVX mice relative to their concentrations in sham-operated mice. In in vitro studies, 17β-estradiol (E(2)) increased the mRNA expression of hepcidin in HepG2 cells in a concentration-dependent manner. E(2)-induced hepatic hepcidin upregulation was not inhibited by ICI 182720, an inhibitor of the estrogen receptor; instead, hepcidin expression was increased by ICI 182720. E(2) and ICI 182720 exhibit agonist actions with G-protein coupled receptor 30 (GPR30), the 7-transmembrane estrogen receptor. G1, a GPR30 agonist, upregulated hepcidin expression, and GPR30 siRNA treatment abolished E(2)-induced hepcidin expression. BMP6 expression induced by E(2) was abolished by GPR30 silencing. Finally, both E(2) and G1 supplementation restored reduced hepatic hepcidin and BMP6 expression and reversed the augmentation of duodenal FPN expression in the OVX mice. In contrast, serum hepcidin was elevated in OVX mice, which was reversed in these mice with E(2) and G1. Thus, estrogen is involved in hepcidin expression via a GPR30-BMP6-dependent mechanism, providing new insight into the role of estrogen in iron metabolism.


The "inside out" transforaminal technique to treat lumbar spinal pain in an awake and aware patient under local anesthesia: results and a review of the literature.
Surgical management of back and leg pain is evolving and changing due to a better understanding of the patho-anatomy well correlated with its pathophysiology. Pain is better understood with in vivo visualization and probing of the pain generators using an endoscopic access rather than just relying on symptoms diagram and image correlation. This has resulted in a shared decision making involving patient and surgeon, focused on a broader spectrum of surgical as well as non-surgical treatments, and not just masking the pain generator. It has moved away from decisions based on diagnostic images alone, that, while noting the image alterations, cannot explain the pain experienced by each individual as images do not always show variations in nerve supply and patho-anatomy. The ability to isolate and visualize "pain" generators in the foramen and treating persistent pain by visualizing inflammation and compression of nerves, serves as the basis for transforaminal endoscopic (TFE) surgery. This has also resulted in better pre surgical planning with more specific and defined goals in mind. The "Inside out" philosophy of TFE surgery is safe and precise. It provides basic access to the disc and foramen to cover a large spectrum of painful pathologies.


Birth Weight, Birth Length, and Gestational Age as Indicators of Favorable Fetal Growth Conditions in a US Sample.
The "fetal origins" hypothesis suggests that fetal conditions not only affect birth characteristics such as birth weight and gestational age, but also have lifelong health implications. Despite widespread interest in this hypothesis, few methodological advances have been proposed to improve the measurement and modeling of fetal conditions. A Statistics in Medicine paper by Bollen, Noble, and Adair examined favorable fetal growth conditions (FFGC) as a latent variable. Their study of Filipino children from Cebu provided evidence consistent with treating FFGC as a latent variable that largely mediates the effects of mother's characteristics on birth weight, birth length, and gestational age. This innovative method may have widespread utility, but only if the model applies equally well across diverse settings. Our study assesses whether the FFGC model of Cebu replicates and generalizes to a very different population of children from North Carolina (N=705) and Pennsylvania (N=494). Using a series of structural equation models, we find that key features of the Cebu analysis replicate and generalize while we also highlight differences between these studies. Our results support treating fetal conditions as a latent variable when researchers test the fetal origins hypothesis. In addition to contributing to the substantive literature on measuring fetal conditions, we also discuss the meaning and challenges involved in replicating prior research.


The inter-rater and test-retest reliability of the Self-care and Transfer scales, and intra-rater reliability of all scales of the Swedish Translation of the Australian Therapy Outcome Measures for Occupational Therapy (AusTOMs-OT-S).
The Australian Therapy Outcome Measures for Occupational Therapy (AusTOMs-OT) are used internationally to measure the outcomes of occupational therapy services across diagnoses, ages, and health care settings. This study reports on the inter-rater and test-retest reliability of the Self-care and Transfer scales as well as the intra-rater reliability of all scales of the Swedish (AusTOMs-OT-S) translation. Fifteen occupational therapists rated 11 case study clients on two occasions, separated by two weeks. Test-retest reliability and inter-rater reliability were calculated for the Self-care scale and Transfer scales. Moreover, intra-rater reliability was calculated for each of the 15 therapists across all 12 scales. The inter-rater reliability intraclass correlation coefficients (ICCs) were all found to be high to very high, ranging from ICC 0.762 to 0.904; the intra-rater reliability coefficients were also very good with 11 of the 15 therapists achieving ICCs of 0.745 or over, and finally the test-retest ICCs were also found to be high, ranging from 0.705 to 0.920. Although further research is required to confirm reliability, preliminary reliability of the AusTOMs-OT Swedish translation has been demonstrated and therapists can have confidence when using the scales.


[Burn-out, commitment, personality and experiences during work and training; survey among psychiatry residents].
In the last few years international studies have reported on increase in burn-out and depressive symptoms among psychiatry residents. In the field of research, however, commitment and dedication are now being mentioned more frequently as positive factors that counterbalance burn-out. To find out how a group of Dutch psychiatry residents feel about their work, to discover their degree of burn-out and commitment and to clarify the various factors involved. 59 psychiatry residents from four teaching hospitals were asked to complete questionnaires concerning burn-out (U-BOS-C), commitment (UWES-15) and personality (BFI-NL). Respondents were also asked to describe how they felt about their experiences during their work and to give their views on the instruction and training they were receiving. In the U-BOS-C section only four trainees (almost 7%) met the criteria for burn-out. In the BFI-NL section the psychiatry residents obtained significantly lower scores on neuroticism and higher scores on empathy than did a comparable norm group of a similar age. The scores of the psychiatry residents indicated that the term 'being proud of your work' was significantly related to a feeling of commitment and particularly to all subscales that reflected commitment. In our study the percentage of psychiatry residents with burn-out is significantly lower than the percentage reported elsewhere in the literature. In fact, our results demonstrate that the psychiatry residents who were the subject of our study regarded themselves as being emotionally stable, friendly and committed to their work.


Regional grey matter structure differences between transsexuals and healthy controls--a voxel based morphometry study.
Gender identity disorder (GID) refers to transsexual individuals who feel that their assigned biological gender is incongruent with their gender identity and this cannot be explained by any physical intersex condition. There is growing scientific interest in the last decades in studying the neuroanatomy and brain functions of transsexual individuals to better understand both the neuroanatomical features of transsexualism and the background of gender identity. So far, results are inconclusive but in general, transsexualism has been associated with a distinct neuroanatomical pattern. Studies mainly focused on male to female (MTF) transsexuals and there is scarcity of data acquired on female to male (FTM) transsexuals. Thus, our aim was to analyze structural MRI data with voxel based morphometry (VBM) obtained from both FTM and MTF transsexuals (n = 17) and compare them to the data of 18 age matched healthy control subjects (both males and females). We found differences in the regional grey matter (GM) structure of transsexual compared with control subjects, independent from their biological gender, in the cerebellum, the left angular gyrus and in the left inferior parietal lobule. Additionally, our findings showed that in several brain areas, regarding their GM volume, transsexual subjects did not differ significantly from controls sharing their gender identity but were different from those sharing their biological gender (areas in the left and right precentral gyri, the left postcentral gyrus, the left posterior cingulate, precuneus and calcarinus, the right cuneus, the right fusiform, lingual, middle and inferior occipital, and inferior temporal gyri). These results support the notion that structural brain differences exist between transsexual and healthy control subjects and that majority of these structural differences are dependent on the biological gender.


Association of Cataract Surgery With Traffic Crashes.
Cataracts are the most common cause of impaired vision worldwide and may increase a driver's risk of a serious traffic crash. The potential benefits of cataract surgery for reducing a patient's subsequent risk of traffic crash are uncertain. To conduct a comprehensive longitudinal analysis testing whether cataract surgery is associated with a reduction in serious traffic crashes where the patient was the driver. Population-based individual-patient self-matching exposure-crossover design in Ontario, Canada, between April 1, 2006, and March 31, 2016. Consecutive patients 65 years and older undergoing cataract surgery (n = 559 546). First eye cataract extraction surgery (most patients received second eye soon after). Emergency department visit for a traffic crash as a driver. Of the 559 546 patients, mean (SD) age was 76 (6) years, 58% were women (n = 326 065), and 86% lived in a city (n = 481 847). A total of 4680 traffic crashes (2.36 per 1000 patient-years) accrued during the 3.5-year baseline interval and 1200 traffic crashes (2.14 per 1000 patient-years) during the 1-year subsequent interval, representing 0.22 fewer crashes per 1000 patient-years following cataract surgery (odds ratio [OR], 0.91; 95% CI, 0.84-0.97; P = .004). The relative reduction included patients with diverse characteristics. No significant reduction was observed in other outcomes, such as traffic crashes where the patient was a passenger (OR, 1.03; 95% CI, 0.96-1.12) or pedestrian (OR, 1.02; 95% CI, 0.88-1.17), nor in other unrelated serious medical emergencies. Patients with younger age (OR, 1.27; 95% CI, 1.13-1.14), male sex (OR, 1.64; 95% CI, 1.46-1.85), a history of crash (baseline OR, 2.79; 95% CI, 1.94-4.02; induction OR, 4.26; 95% CI, 2.01-9.03), more emergency visits (OR, 1.34; 95% CI, 1.19-1.52), and frequent outpatient physician visits (OR, 1.17; 95% CI, 1.01-1.36) had higher risk of subsequent traffic crashes (multivariable model). This study suggests that cataract surgery is associated with a modest decrease in a patient's subsequent risk of a serious traffic crash as a driver, which has potential implications for mortality, morbidity, and costs to society.


Nonstationary chaotic oscillations in lasers with frequency-shifted feedback.
The nonlinear dynamics of lasers with frequency-shifted delayed feedback are investigated. Resonant excitation of sustained relaxation oscillations by harmonic resonance is demonstrated. Self-induced switching between sustained relaxation oscillation and spiking oscillation is observed as the feedback coefficient is increased. Observed instabilities are well reproduced by numerical simulations of proposed model equations. A statistical analysis of this switching phenomenon is carried out numerically, and the results indicate that an inverse-power relation with the feedback coefficient determines the periods over which the system dwells in its relaxation-oscillation state.


Thermal drying of wastewater sludge with crack formation.
We examined in this work the drying characteristics of wastewater sludge. The drying flux of the constant-rate period for sludge cake could be up to 40% higher than that from a sand bed. Owing to the considerable volume shrinkage of cake, cracks would form and develop on the crack surface, which yielded three-dimensional but rather than the one-dimensional cake structure assumed in conventional drying theories. The crack length was fully developed in the first 30 min of drying, while the width of crack increased linearly with time. Using these data the drying flux from the cracks was estimated. Enhanced drying flux was noticeable which depended on the crack shape and the crossflow velocity.


Genome Analysis of Vallitalea guaymasensis Strain L81 Isolated from a Deep-Sea Hydrothermal Vent System.
Abyssivirga alkaniphila strain L81T, recently isolated from a black smoker biofilm at the Loki's Castle hydrothermal vent field, was previously described as a mesophilic, obligately anaerobic heterotroph able to ferment carbohydrates, peptides, and aliphatic hydrocarbons. The strain was classified as a new genus within the family Lachnospiraceae. Herein, its genome is analyzed and A. alkaniphila is reassigned to the genus Vallitalea as a new strain of V. guaymasensis, designated V. guaymasensis strain L81. The 6.4 Mbp genome contained 5651 protein encoding genes, whereof 4043 were given a functional prediction. Pathways for fermentation of mono-saccharides, di-saccharides, peptides, and amino acids were identified whereas a complete pathway for the fermentation of n-alkanes was not found. Growth on carbohydrates and proteinous compounds supported methane production in co-cultures with Methanoplanus limicola. Multiple confurcating hydrogen-producing hydrogenases, a putative bifurcating electron-transferring flavoprotein—butyryl-CoA dehydrogenase complex, and a Rnf-complex form a basis for the observed hydrogen-production and a putative reverse electron-transport in V. guaymasensis strain L81. Combined with the observation that n-alkanes did not support growth in co-cultures with M. limicola, it seemed more plausible that the previously observed degradation patterns of crude-oil in strain L81 are explained by unspecific activation and may represent a detoxification mechanism, representing an interesting ecological function. Genes encoding a capacity for polyketide synthesis, prophages, and resistance to antibiotics shows interactions with the co-occurring microorganisms. This study enlightens the function of the fermentative microorganisms from hydrothermal vents systems and adds valuable information on the bioprospecting potential emerging in deep-sea hydrothermal systems.


Molecular Hydrogen Metabolism: a Widespread Trait of Pathogenic Bacteria and Protists.
Pathogenic microorganisms use various mechanisms to conserve energy in host tissues and environmental reservoirs. One widespread but often overlooked means of energy conservation is through the consumption or production of molecular hydrogen (H2). Here, we comprehensively review the distribution, biochemistry, and physiology of H2 metabolism in pathogens. Over 200 pathogens and pathobionts carry genes for hydrogenases, the enzymes responsible for H2 oxidation and/or production. Furthermore, at least 46 of these species have been experimentally shown to consume or produce H2 Several major human pathogens use the large amounts of H2 produced by colonic microbiota as an energy source for aerobic or anaerobic respiration. This process has been shown to be critical for growth and virulence of the gastrointestinal bacteria Salmonella enterica serovar Typhimurium, Campylobacter jejuni, Campylobacter concisus, and Helicobacter pylori (including carcinogenic strains). H2 oxidation is generally a facultative trait controlled by central regulators in response to energy and oxidant availability. Other bacterial and protist pathogens produce H2 as a diffusible end product of fermentation processes. These include facultative anaerobes such as Escherichia coli, S Typhimurium, and Giardia intestinalis, which persist by fermentation when limited for respiratory electron acceptors, as well as obligate anaerobes, such as Clostridium perfringens, Clostridioides difficile, and Trichomonas vaginalis, that produce large amounts of H2 during growth. Overall, there is a rich literature on hydrogenases in growth, survival, and virulence in some pathogens. However, we lack a detailed understanding of H2 metabolism in most pathogens, especially obligately anaerobic bacteria, as well as a holistic understanding of gastrointestinal H2 transactions overall. Based on these findings, we also evaluate H2 metabolism as a possible target for drug development or other therapies.


An integrated workflow for phenazine-modifying enzyme characterization.
Increasing availability of new genomes and putative biosynthetic gene clusters (BGCs) has extended the opportunity to access novel chemical diversity for agriculture, medicine, environmental and industrial purposes. However, functional characterization of BGCs through heterologous expression is limited because expression may require complex regulatory mechanisms, specific folding or activation. We developed an integrated workflow for BGC characterization that integrates pathway identification, modular design, DNA synthesis, assembly and characterization. This workflow was applied to characterize multiple phenazine-modifying enzymes. Phenazine pathways are useful for this workflow because all phenazines are derived from a core scaffold for modification by diverse modifying enzymes (PhzM, PhzS, PhzH, and PhzO) that produce characterized compounds. We expressed refactored synthetic modules of previously uncharacterized phenazine BGCs heterologously in Escherichia coli and were able to identify metabolic intermediates they produced, including a previously unidentified metabolite. These results demonstrate how this approach can accelerate functional characterization of BGCs.


Ready-made matrix-assisted laser desorption/ionization target plates coated with thin matrix layer for automated sample deposition in high-density array format.
The methodology for ready-made matrix-assisted laser desorption/ionization (MALDI) target plates covered with an optimized thin layer consisting of matrix and nitrocellulose has been developed. Piezoelectric microdispensing enabled sample depositions in a high-density array format of 2000 sample depositions on a conventionally sized target plate (45 x 47 mm). The sample depositions were made reproducibly in a fully automated mode by using an in-house developed computer-controlled piezoelectric flow-through microdispenser. Additionally, the piezoelectric technique facilitated significant analyte enrichment that increased the detection sensitivity. The MS signal was obtained rapidly, generally within ten laser pulses. An airbrush device was used to generate a fine spray of matrix and nitrocellulose dissolved in acetone. The acetone evaporated instantly when reaching the target plate leaving the entire surface with a thin and uniform matrix/nitrocellulose coating consisting of very small crystals of matrix embedded in the nitrocellulose. These crystals acted as a seed-layer on subsequent analyte depositions, rendering homogeneous sample spots when using alpha-cyano-4-hydroxycinnamic acid (CHCA) as matrix. The relative standard deviation of the signal intensity between spots was (20-30)% (n = 30). The detection sensitivity was improved by restricting the sample spot diameter to 300 microm. The spot size was affected by the deposition rate and the evaporation rate of the dispensed sample volume. Mass spectra of a 25-amol peptide mixture deposition were successfully recorded.


Structures, electronic properties and stability phase diagrams for copper(I/II) bromide surfaces.
This study presents a comprehensive periodic slab DFT investigation into structures, electronic properties and thermodynamic stability of all plausible terminations of CuBr and CuBr2 surfaces. We first estimate lattice constants, formation and cohesive energies for the two bulk copper bromides before proceeding to analyse geometrical and electronic features of CuBr and CuBr2 configurations. Surface geometries exhibit, to a large extent, corresponding bulk structures. Nevertheless, certain CuBr2 surfaces experience a downward displacement of the topmost Cu-containing layers. We plot total and projected density of states for bulk and surface geometries of these two copper bromides and calculate their associated Bader's electronic charges. Electronic structure analysis for the bulk and surfaces of these two copper bromides show that CuBr bulk and its most stable surface (CuBr(001)_Br) do not exhibit any metallic character, whereas CuBr2 bulk and its most stable surface (CuBr2(001)_Br) both exhibit metallic characters. The formalism of the ab initio atomistic thermodynamics affords the construction of energy phase diagrams. We predict that the CuBr(001) surface, truncated with Br atoms, is the most stable structure among the considered CuBr slabs at all physically meaningful ranges of the chemical potential of bromine. This surface resembles a c(2 × 2)-bromine sheet that was characterised experimentally from initial interaction of Br2 with a Cu(100) surface. We find that surfaces terminated with the electronegative bromine atoms, if accompanied by significant relaxation, tend to be more stable. Calculated surface energies predict the shapes of CuBr and CuBr2 nanoparticles as the chemical potential of bromine changes.