Dopamine and gamma band synchrony in schizophrenia--insights from computational and empirical studies.
Dopamine modulates cortical circuit activity in part through its actions on GABAergic interneurons, including increasing the excitability of fast-spiking interneurons. Though such effects have been demonstrated in single cells, there are no studies that examine how such mechanisms may lead to the effects of dopamine at a neural network level. With this motivation, we investigated the effects of dopamine on synchronization in a simulated neural network composed of excitatory and fast-spiking inhibitory Wang-Buzsaki neurons. The effects of dopamine were implemented through varying leak K+ conductance of the fast-spiking interneurons and the network synchronization within the gamma band (∼40 Hz) was analyzed. Parametrically varying the leak K+ conductance revealed an inverted-U shaped relationship, with low gamma band power at both low and high conductance levels and optimal synchronization at intermediate conductance levels. We also examined the effects of modulating excitability of the inhibitory neurons more generically using an idealized model with theta neurons, with similar findings. Moreover, such a relationship holds when the external input is both tonic and periodic. Our computational results mirror our empirical study of dopamine modulation in schizophrenia and healthy controls, which showed that amphetamine administration increased gamma power in patients but decreased it in controls. Together, our computational and empirical investigations indicate that dopamine can modulate cortical gamma band synchrony in an inverted-U fashion and that the physiologic effects of dopamine on single fast-spiking interneurons can give rise to such non-monotonic effects at the network level.


Chilean propolis: antioxidant activity and antiproliferative action in human tumor cell lines.
Propolis, a natural product derived from plant resins collected by honeybees, has been used for thousands of years in traditional medicine all over the world. The composition of the propolis depends upon the vegetation of the area from where it was collected and on the bee species. In this study, we investigated the antioxidant activity of a propolis sample, provided by NATURANDES-CHILE, collected in a temperate region of central Chile. In addition, this natural compound was tested for its antiproliferative capacity on KB (human mouth epidermoid carcinoma cells), Caco-2 (colon adenocarcinoma cells) and DU-145 (androgen-insensitive prostate cancer cells) human tumor cell lines. Results showed that this Chilean propolis sample exhibits interesting biological properties, correlated with its chemical composition and expressed by its capacity to scavenge free radicals and to inhibit tumor cell growth.


Tuning hydrogels through metal-based gelation triggers.
Two N-terminal capped tripeptides have been synthesised and display remarkable versatility in their gelation behaviour. Here, gelation can be triggered through the addition of an acid, biologically relevant metal salts or cell culture media. The structural and mechanical properties of the resultant hydrogels are tuneable, with gelation triggered through sodium cations amplifying secondary structure organisation yet decreasing gel stiffness. The effect of gelation trigger on cell viability is evaluated, with substantial improvement in cell survival observed when gelation is triggered using metal salts or cell culture media.


Yttrium-90 radioembolization for liver malignancies: prognostic factors associated with survival.
To identify key prognostic clinical and imaging variables in patients undergoing yttrium-90 radioembolization ((90)Y) for liver malignancies. Patients with liver malignancies that progressed despite standard-of-care therapy were treated with (90)Y from 2002 to 2006. Baseline functional status, laboratory values, and diagnostic imaging were assessed before therapy. Imaging follow-up was performed 1 month after treatment and subsequently at 3-month intervals. Patients were followed for survival from the time of their first (90)Y treatment. Patients with follow-up imaging after radioembolization (N = 130) were included in this analysis. Primary malignancies included colon, neuroendocrine, and others. The following clinical variables had a significant effect on survival on multivariate analysis: Eastern Cooperative Oncology Group (ECOG) performance status (PS) greater than 0 (hazard ratio [HR], 7.98; 95% CI, 3.98-16), hepatic tumor burden of 51%-75% (HR, 2.46; 95% CI, 1.01-6.02), bilirubin level greater than 1.3 mg/dL (HR, 2.60; 95% CI, 1.27-5.34), hepatic metastases from breast cancer (HR, 2.51; 95% CI, 1.13-5.61), response on imaging based on World Health Organization (WHO) criteria (HR, 0.48; 95% CI, 0.24-0.94), and lymphocyte depression (HR, 0.56; 95% CI, 0.31-0.96). Among patients with colorectal cancer metastases to the liver, the HR for survival on univariate analysis for responders compared with nonresponders (per WHO criteria) was 0.26 (95% CI, 0.10-0.69). Cancer-related symptoms (ie, ECOG PS > 0), hepatic tumor burden greater than 50%, increased bilirubin levels, and hepatic metastases from breast cancer were found to be negative prognostic factors. Tumor response to therapy and lymphocyte depression were associated with favorable prognosis. Additionally, WHO response was identified to be a favorable prognostic factor in patients with colorectal cancer metastases. These findings may be useful when counseling patients regarding prognosis of their hepatic disease.


Cardiovascular effects of dipeptidyl peptidase-4 inhibitors in patients with type 2 diabetes.
Cardiovascular (CV) disease is the leading cause of mortality and morbidity in patients with type 2 diabetes mellitus (T2DM). However, improving glycaemic control alone has not decreased CV events. Therapies that improve glycaemic control, CV disease risk factors and CV function are more likely to be successful. Dipeptidyl peptidase-4 (DPP-4) inhibitors prevent breakdown of incretin hormones glucagon-like peptide-1(GLP-1) and glucose-dependent insulinotropic peptide and improve glycaemic control in patients with T2DM. DPP-4 acts on other substrates, many associated with cardioprotection. Thus, inhibition of DPP-4 may lead to elevations in these potentially beneficial substrates. Data from animal studies and small observational studies in humans suggest that DPP-4 inhibitors may potentially reduce CV risk. However, recently completed CV outcome trials in patients with T2DM and CV disease or at high risk of adverse CV events have shown that the DPP-4 inhibitors saxagliptin and alogliptin neither increased nor decreased major adverse CV events.


Comparative proteomics of Pinus-Fusarium circinatum interactions reveal metabolic clues to biotic stress resistance.
Fusarium circinatum, causing pine pitch canker (PPC), affects conifers productivity and health worldwide. Selection and breeding for resistance arises as the most promising approach to fight PPC. Therefore, it is crucial to explore the response of hosts with varying levels of susceptibility to PPC to unveil the genes/pathways behind these phenotypes. We evaluated the dynamics of the needle proteome of a susceptible (Pinus radiata) and a relatively resistant (Pinus pinea) species upon F. circinatum inoculation by GeLC-MS/MS. Integration with physiological data and validation of key genes by qPCR allowed to identify core pathways regulating these contrasting responses. In P. radiata, the pathogen may target both the secondary metabolism to negatively regulate immune response and chloroplast redox proteins to increase energy-producing pathways for amino acid production in its favour. In contrast, chloroplast redox regulation may assure redox homeostasis in P. pinea, as well as nonenzymatic antioxidants. The presence of membrane trafficking-related proteins exclusively in P. pinea likely explains its defence response against F. circinatum. A crosstalk between abscisic acid and epigenetic regulation of gene expression is also proposed in PPC response. These results are useful to support breeding programs aiming to achieve PPC resistance.


Coping with systematic bias during bilateral movement.
The present studies examined the nature of kinematic interlimb interference during bilateral elbow movements of 1:1, 2:1 and 3:1 frequency ratios and the manner in which subjects cope with coordination bias. Analysis of movement trajectories in the first experiment indicated progressively greater angular velocity assimilation across 2:1 and 3:1 conditions. The desired temporal relationship was maintained by slowing or pausing the low-frequency movement at peak extension while the high-frequency arm produced intervening cycles. An increase in amplitude was also evident for concurrent, homologous cycles. Movement smoothness was emphasized and additional practice was provided in a second experiment. This resulted in dissociated peak angular velocity between limbs and eliminated hesitations and amplitude effects. Bias was still evident, however, as an intermittent approach toward a 1:1 ratio within each cycle. This systematic tendency was somewhat greater at the lower of two absolute frequency combinations but was not influenced by the role of each arm in producing the higher or lower frequency movement. The findings from the first experiment suggest that subjects initially accommodate interlimb kinematic assimilation, while producing the intended timing ratio, by intermittently slowing or pausing the lower-frequency movement. This attenuates the need for bilaterally-disparate movement parameters and provides additional time for organizing residual kinematic differences, perhaps reducing "transient coupling." Evidence from the second experiment indicates that subtle relative motion preferences are still evident following sufficient practice to perform the movements smoothly. The within-cycle locations of the points of greatest interlimb bias for the 2:1 rhythms were positively displaced from those previously observed for 1:1 oscillations. The persistent coordination tendencies noted in both experiments perhaps reflect an assimilation/compensation cycle and constitute one potential source of the systematic error that often emerges during the acquisition of complex skills.


The composition of polyunsaturated fatty acids in erythrocytes of lactating mothers and their infants.
Long-chain polyunsaturated fatty acids (LCPUFA) in breastmilk, specifically docosahexaenoic acid (DHA), are important for infant brain development. Accretion of DHA in the infant brain is dependent on DHA-status, intake and metabolism. The aim of this study was to describe changes in maternal and infant erythrocyte (RBC) DHA-status during the first four months of lactation. We examined 17 mothers and their term infants at 1, 2 and 4 months of age. Milk samples and RBC from the mothers and infants were obtained and analysed for fatty acid composition. Comparative analysis of the results showed that the content of DHA in maternal RBC-phosphatidylcholine (PE) decreased over the four month period and this was not accompanied by a decrease in DHA in infant RBC-PE (P = 0.005). The ratio of n-6 PUFA to n-3 PUFA increased over time in maternal RBC-PE, but not in infant RBC-PE (P < 0.001). The level of 22:5n-6 and the ratio of LCPUFA to precursor PUFAs in infant RBC was higher than in maternal RBC phospholipids. (P = and P < 0.001 respectively). We found a decrease in the level of LCPUFA in milk, specifically AA. However, we did not observe a significant decrease in milk DHA, which may have been due to two outliers. These results indicate better DHA-status and a higher n-3/n-6 PUFA in RBC of infants than in mothers. Whether these differences reflect preferential n-3 PUFA transfer via breastmilk or differences in PUFA-metabolism and utilization remains to be shown.


Keeping abreast of shark bite reconstruction: a case report and brief literature review.
Shark attack-related injuries (SARIs) are rare but may result in highly complex wounds requiring reconstruction by plastic surgeons. Here, we present an unusual case of SARI of the breast and briefly review literature on reconstructive management of (1) breast injuries from other large animals and (2) SARIs to other parts of the body. Our patient was a 39-year-old woman who experienced massive bilateral breast tissue loss from a shark attack. After primary surgical wound management, including debridement and washout, the patient underwent completion mastectomy with bilateral deep inferior epigastric perforator flap breast reconstruction. Literature review revealed that reports of animal-related injuries to the breast are rare, with ours being the first on SARI to the breast and the only describing major reconstruction of animal-related breast injury. Literature on reconstruction of non-breast SARI wounds is limited to two case reports describing severe tissue damage akin to that seen in our patient, both necessitating extensive debridement followed by reconstruction. Overall, our findings demonstrate the uniquely devastating damage resulting from SARIs and the value of primary wound management and abdominally based free flaps in successfully reconstructing these wounds. Level of evidence: Level V, therapeutic study.


[Evaluation of the pain-alleviating properties of a combination of analgesic and tranquilizer using multiple stepwise regression].
A method for preclinical evaluation of the optimal ratio (for analgesia) of an analgesic to a tranquilizer has been devised. It is based on the drug threshold dose decrease on the drug combined use. The multiple stepwise regression analysis of the data discovered the correlations between diazepam and promedol, optimal for pain perception decrease and nociceptive emotional responsiveness inhibition.


Mycophenolate blood level monitoring: recent progress.
The concentration-effect relationship for mycophenolic acid (MPA), and the high variability in MPA concentrations in patients on standard dose mycophenolate mofetil (MMF) therapy, for some centers has provided enough evidence to implement therapeutic drug monitoring (TDM) for MMF in daily practice. Two randomized trials Adaption de Posologie du MMF en Greffe Renale (APOMYGRE) and fixed-dose versus concentration controlled (FDCC) investigated the added benefit of TDM for MMF in renal transplant recipients. The APOMYGRE study showed a significant reduction in the incidence of acute rejection in concentration-controlled patients, while the FDCC study had a negative outcome, despite a similar study design. Although it was expected that these prospective trials would give the final answer to the question of whether or not TDM for MMF would be of benefit, it seems that the studies have not had much impact on patient management. Several trials have shown the importance of early adequate exposure to MPA in the first week after transplantation. As it will be hard to improve MPA exposure with TDM, this early, ongoing study now investigates the use of an increased starting dose. The increased starting dose will avoid underexposure to MPA in higher proportions of patients shortly after transplantation but may result in more toxicity in patients with MPA exposures exceeding the upper threshold of the therapeutic window.


Influenza A virus polymerase inhibits type I interferon induction by binding to interferon beta promoter stimulator 1.
Type I interferons (IFNs) are known to be critical factors in the activation of host antiviral responses and are also important in protection from influenza A virus infection. Especially, the RIG-I- and IPS-1-mediated intracellular type I IFN-inducing pathway is essential in the activation of antiviral responses in cells infected by influenza A virus. Previously, it has been reported that influenza A virus NS1 is involved in the inhibition of this pathway. We show in this report that the influenza A virus utilizes another critical inhibitory mechanism in this pathway. In fact, the viral polymerase complex exhibited an inhibitory activity on IFNβ promoter activation mediated by RIG-I and IPS-1, and this activity was not competitive with the function of NS1. Co-immunoprecipitation analysis revealed that each polymerase subunit bound to IPS-1 in mammalian cells, and each subunit inhibited the activation of IFNβ promoter by IPS-1 independently. In addition, by a combinational expression of each polymerase subunit, IPS-1-induced activation of IFNβ promoter was more efficiently inhibited by the expression of PB2 or PB2-containing complex. Moreover, the expression of PB2 inhibited the transcription of the endogenous IFNβ gene induced after influenza A virus infection. These findings demonstrate that the viral polymerase plays an important role for regulating host anti-viral response through the binding to IPS-1 and inhibition of IFNβ production.


Legal medicine--what general practitioners should know?
General practitioners are the nearest to the community who can render the need of the patient both clinically and physiologically. A general practitioner is to develop the art of communication to avoid litigation and be aware of all the legal issues entangling the profession so that his practice is not interfered by the law enforcing authority. A general practitioner does not have the time to update his knowledge on legal issues involved in practice and this articles lends a few input for them.


Super-Memorizers Are Not Super-Recognizers.
Humans have a natural expertise in recognizing faces. However, the nature of the interaction between this critical visual biological skill and memory is yet unclear. Here, we had the unique opportunity to test two individuals who have had exceptional success in the World Memory Championships, including several world records in face-name association memory. We designed a range of face processing tasks to determine whether superior/expert face memory skills are associated with distinctive perceptual strategies for processing faces. Superior memorizers excelled at tasks involving associative face-name learning. Nevertheless, they were as impaired as controls in tasks probing the efficiency of the face system: face inversion and the other-race effect. Super memorizers did not show increased hippocampal volumes, and exhibited optimal generic eye movement strategies when they performed complex multi-item face-name associations. Our data show that the visual computations of the face system are not malleable and are robust to acquired expertise involving extensive training of associative memory.


MRI kinetics with volumetric analysis in correlation with hormonal receptor subtypes and histologic grade of invasive breast cancers.
OBJECTIVE. The aim of this study was to assess whether computer-assisted detection-processed MRI kinetics data can provide further information on the biologic aggressiveness of breast tumors. MATERIALS AND METHODS. We identified 194 newly diagnosed invasive breast cancers presenting as masses on contrast-enhanced MRI by a HIPAA-compliant pathology database search. Computer-assisted detection-derived data for the mean and median peak signal intensity percentage increase, most suspicious kinetic curve patterns, and volumetric analysis of the different kinetic patterns by mean percentage tumor volume were compared against the different hormonal receptor (estrogen-receptor [ER], progesterone-receptor [PR], ERRB2 (HER2/neu), and triple-receptor expressivity) and histologic grade subgroups, which were used as indicators of tumor aggressiveness. RESULTS. The means and medians of the peak signal intensity percentage increase were higher in ER-negative, PR-negative, and triple-negative (all p ≤ 0.001), and grade 3 tumors (p = 0.011). Volumetric analysis showed higher mean percentage volume of rapid initial enhancement in biologically more aggressive ER-negative, PR-negative, and triple-negative tumors compared with ER-positive (64% vs 53.6%, p = 0.013), PR-positive (65.4% vs 52.5%, p = 0.001), and nontriple-negative tumors (65.3% vs 54.6%, p = 0.028), respectively. A higher mean percentage volume of rapid washout component was seen in ERRB2-positive tumors compared with ERRB2-negative tumors (27.5% vs 17.9%, p = 0.020). CONCLUSION. Peak signal intensity percentage increase and volume analysis of the different kinetic patterns of breast tumors showed correlation with hormonal receptor and histologic grade indicators of cancer aggressiveness. Computer-assisted detection-derived MRI kinetics data have the potential to further characterize the aggressiveness of an invasive cancer.


The serine protease HtrA1 is upregulated in the human placenta during pregnancy.
The placenta has a dynamic and continuous capacity for self-renewal. The molecular mechanisms responsible for controlling trophoblast proliferation are still unclear. It is generally accepted that the simultaneous activity of proteins involved in cell proliferation, apoptosis, and extracellular matrix degradation plays an important role in correct placental development. We investigated in depth the expression of the serine protease HtrA1 during pregnancy in human placenta by in situ hybridization and immunohistochemistry, we demonstrated that HtrA1 displayed a low level of expression in the first trimester of gestation and a strong increase of HtrA1 expression in the third trimester. Finally, by electron microscopy, we demonstrated that HtrA1 was localized either in the cytoplasm of placental cells, especially close to microvilli that characterized the plasma membrane of syncytiotrophoblast cells, or in the extracytoplasmic space of the stroma of placental villi, particularly in the spaces between collagen fibers and on collagen fibers themselves. The expression pattern of HtrA1 in human placentas strongly suggests a role for this protein in placental development and function. Moreover, on the basis of its subcellular distribution it can be postulated that HtrA1 acts on different targets, such as intracellular growth factors or extracellular matrix proteins, to favor the correct formation/function of the placenta.


Evaluation of cowpea mini core accessions for resistance to flower bud thrips Megalurothrips sjostedti Trybom (Thysanoptera: Thripidae).
The flower bud thrips, Megalurothrips sjostedti Trybom (Thysanoptera: Thripidae), is an economically important pest of cowpea in sub-Saharan Africa. Varietal resistance is the most preferred, environmentally friendly, cost-effective and sustainable option for controlling this pest. The objective of this study was to identify sources of resistance to M. sjostedti among mini core accessions from the largest world cowpea germplasm collection maintained at the International Institute of Tropical Agriculture (IITA). The study was conducted during the 2015 and 2016 cropping seasons where 365 accessions were screened under field conditions. Each accession was rated visually for thrips damage score, flower abortion rate, number of pods per plant and number of thrips per flower. The resistance levels observed in genotypes TVu8631, TVu16368, TVu8671 and TVu7325 were similar to that of the resistant check "Sanzisabinli" (called Sanzi) during both seasons. In addition, 56 mini core genotypes showed moderate resistance to thrips damage. High heritability values were associated with thrips damage scores at 65 days after planting (0.60), percentage of effective peduncles (0.59), flower bud abortion rate (0.59), number of pods per plant (0.51) and number of peduncles with pods (0.5). The accessions identified with good levels of resistance to flower bud thrips will be used in cowpea breeding programs to develop improved resistant varieties.


Alterations of Na+-K+-ATPase, Ca2+-ATPase, and Mg2+-ATPase activities in erythrocyte, muscle, and liver of traumatic and septic patients.
Na+-K+-ATPase, Ca2+-ATPase and Mg2+-ATPase activities of erythrocyte membrane, microsomal fractions of rectus muscle, and liver were measured colorimetrically in the biopsy specimens of 14 control, 7 uncomplicated trauma (group 2), and 14 severe trauma or septic patients (groups 3-A and 3-B). In erythrocytes, these three ATPase activities in group 2 were not significantly changed but sepsis of both the acute (group 3-A) and ongoing type (group 3-B) decreased all of the ATPase activities. In muscle, there was a significant loss of three ATPase activities in the acute insult of severe trauma or sepsis (group 3-A), while Na+-K+-ATPase and Mg2+-ATPase activities were not significantly changed in ongoing, severe trauma (group 3-B). In the liver, a tendency for all three ATPase activities to decrease is noted in the severe traumatic group. However, a statistical difference between the control and severe traumatic group showed only for Na+-K+ ATPase and Mg2+-ATPase in group 3-A and Ca2+-ATPase in group 3-B. Correlation coefficients between erythrocyte, muscle, and liver for three ATPase activities are between 0.4 and 0.5. The mechanism which alters ATPase activity remains unknown in this study, but it may account for the variation in traumatic insult, in hemodynamic and hormone changes, and in tissue energy stores.


SPECTRAL DOMAIN OPTICAL COHERENCE TOMOGRAPHY FINDINGS IN MACULA-INVOLVING CYTOMEGALOVIRUS RETINITIS.
To evaluate the microstructural features of cytomegalovirus (CMV) retinitis by spectral domain optical coherence tomography (OCT). Subjects were patients with macula-involving CMV retinitis with OCT imaging. The leading edge of retinitis in the macula was identified based on fundus imaging, and OCT findings were longitudinally evaluated in three areas: within the area of active retinitis, at the leading edge of retinitis, and just beyond the leading edge of retinitis. Optical coherence tomography imaging of macular CMV retinitis identified vitreous cells in 10 eyes (100%), posterior vitreous detachment in four eyes (40%), broad-based vitreomacular traction in one eye (10%), epiretinal membrane in eight eyes (80%), and lamellar hole-associated epiretinal proliferation associated with an atrophic hole in one eye (10%). Retinal architectural disruption, disruption of inner retinal layers, disruption of the external limiting membrane, and ellipsoid zone abnormalities were noted within the area of retinitis in all eyes and decreased in frequency and severity at and beyond the leading edge of retinitis, although all 10 eyes (100%) exhibited one of these abnormalities, especially outer retinal microabnormalities, beyond the leading edge of retinitis. Microstructural abnormalities were frequently noted on OCT of CMV retinitis, including within the retina beyond the leading edge of retinitis identified by corresponding fundus imaging. Outer retinal abnormalities were noted more frequently than inner retinal abnormalities beyond the leading edge of retinitis. These findings provide insight into the effects of CMV retinitis on retinal microstructure and potentially on vision and highlight the potential utility of OCT for monitoring microprogression of macula-involving CMV retinitis.


Natural Polysaccharide Carriers in Brain Delivery: Challenge and Perspective.
Targeted drug delivery systems represent valuable tools to enhance the accumulation of therapeutics in the brain. Here, the presence of the blood brain barrier strongly hinders the passage of foreign substances, often limiting the effectiveness of pharmacological therapies. Among the plethora of materials used for the development of these systems, natural polysaccharides are attracting growing interest because of their biocompatibility, muco-adhesion, and chemical versatility which allow a wide range of carriers with tailored physico-chemical features to be synthetized. This review describes the state of the art in the field of targeted carriers based on natural polysaccharides over the last five years, focusing on the main targeting strategies, namely passive and active transport, stimuli-responsive materials and the administration route. In addition, in the last section, the efficacy of the reviewed carriers in each specific brain diseases is summarized and commented on in terms of enhancement of either blood brain barrier (BBB) permeation ability or drug bioavailability in the brain.


Analysis of the PAX8 gene in congenital hypothyroidism caused by different forms of thyroid dysgenesis in a father and daughter.
Thyroid dysgenesis occurs sporadically with only rare familial presentation. We report a father and daughter with congenital hypothyroidism caused by different forms of thyroid dysgenesis. The father had a severely hypoplastic thyroid gland in a normal location, whereas the daughter had an ectopic thyroid gland in a sublingual position. Her brother had a hypoplastic thyroid but was euthyroid. The involvement of the candidate gene, PAX8, as the cause of thyroid dysgenesis in this family was partially excluded by linkage analysis, and the possibility of a de novo mutation excluded by sequencing.


The tumor suppressive effect and apoptotic mechanism of TRAIL gene-containing recombinant NDV in TRAIL-resistant colorectal cancer HT-29 cells and TRAIL-nonresistant HCT116 cells, with each cell bearing a mouse model.
TRAIL is an anticancer drug that induces cancer cell apoptosis by interacting with death receptors (DRs). However, owing to low cell-surface expression of DRs, certain colorectal cancer (CRC) cells resist TRAIL-induced apoptosis. Newcastle disease virus (NDV) infection can elevate DR protein expression in cancer cells, potentially influencing their TRAIL sensitivity. However, the precise mechanism by which NDV infection modulates DR expression and impacts TRAIL sensitivity in cancer cells remains unknown. Herein, we developed nonpathogenic NDV VG/GA strain-based recombinant NDV (rNDV) and TRAIL gene-containing rNDV (rNDV-TRAIL). We observed that viral infections lead to increased DR and TRAIL expressions and activate signaling proteins involved in intrinsic and extrinsic apoptosis pathways. Experiments were conducted in vitro using TRAIL-resistant CRC cells (HT-29) and nonresistant CRC cells (HCT116) and in vivo using relevant mouse models. rNDV-TRAIL was found to exhibit better apoptotic efficacy than rNDV in CRC cells. Notably, rNDV-TRAIL had the stronger cancer cell-killing effect in TRAIL-resistant CRC cells. Western blot analyses showed that both rNDV and rNDV-TRAIL infections activate signaling proteins involved in the intrinsic and extrinsic apoptotic pathways. Notably, rNDV-TRAIL promotes concurrent intrinsic and extrinsic signal transduction in both HCT-116 and HT-29 cells. Therefore, rNDV-TRAIL infection effectively enhances DR expression in DR-depressed HT-29 cells. Moreover, the TRAIL protein expressed by rNDV-TRAIL effectively interacts with DR, leading to enhanced apoptosis in TRAIL-resistant HT-29 cells. Therefore, rNDV-TRAIL has potential as a promising therapeutic approach for treating TRAIL-resistant cancers.


Effect of Oral Zinc Supplementation on Serum Bilirubin Levels in Term Neonates With Hyperbilirubinemia Undergoing Phototherapy: A Double-blind Randomized Controlled Trial.
Background: Enterohepatic bilirubin circulation is one of the determinants of neonatal jaundice. To evaluate the role of oral zinc in reducing serum bilirubin in term neonates with hyperbilirubinemia. Double-blind, randomized, placebo-controlled trial. 106 term neonates with jaundice within the phototherapy range admitted to a level III neonatal intensive care unit. Neonates were randomized and allocated to receive either oral zinc sulfate (5 mg/day) or matching placebo for 5 days. Both groups received conventional phototherapy as per American Academy of Pediatrics (AAP) guidelines. Primary: Reduction in total serum bilirubin levels at 24, 48, 72, and 96 hr after intervention. Secondary: Duration of phototherapy, and hospital stay. The mean (SD) total serum bilirubin levels in zinc and placebo groups were 15.3 (2.85) vs 17.1 (2.21) mg/dL (MD 1.74; P<0.001) at 24 h; 11.7 (4.46) vs. 14.62 (3.83) mg/dL (MD 2.89; P<0.001) at 48 h; 6.7 (4.77) vs 9.5 (3.70) mg/dL (MD 2.79; P <0.001) at 72 h; and 5.1 (3.95) vs 6.5 (3.70) mg/dL (MD 1,49; P=0.045) after 72 hr, respectively. The mean (SD) duration of phototherapy was significantly lower in zinc group than placebo group [ 53.42 (19.62) vs 71.4 (19.43) h; P<0.001]. There was no significant difference in hospital stay between the two groups [mean (SD) 81.05 (19.43) vs 86.25 (20.02) h; P= 0.227]. Oral zinc sulfate supplementation at a dose of 5 mg once a day along with phototherapy significantly reduced total and indirect serum bilirubin levels and also reduced the total duration of phototherapy required in the term neonatal hyperbilirubinemia, with minimal or no adverse effects.


Early protective effect of a ("pan") coronavirus vaccine (PanCoVac) in Roborovski dwarf hamsters after single-low dose intranasal administration.
The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has highlighted the danger posed by human coronaviruses. Rapid emergence of immunoevasive variants and waning antiviral immunity decrease the effect of the currently available vaccines, which aim at induction of neutralizing antibodies. In contrast, T cells are marginally affected by antigen evolution although they represent the major mediators of virus control and vaccine protection against virus-induced disease. We generated a multi-epitope vaccine (PanCoVac) that encodes the conserved T cell epitopes from all structural proteins of coronaviruses. PanCoVac contains elements that facilitate efficient processing and presentation of PanCoVac-encoded T cell epitopes and can be uploaded to any available vaccine platform. For proof of principle, we cloned PanCoVac into a non-integrating lentivirus vector (NILV-PanCoVac). We chose Roborovski dwarf hamsters for a first step in evaluating PanCoVac in vivo. Unlike mice, they are naturally susceptible to SARS-CoV-2 infection. Moreover, Roborovski dwarf hamsters develop COVID-19-like disease after infection with SARS-CoV-2 enabling us to look at pathology and clinical symptoms. Using HLA-A*0201-restricted reporter T cells and U251 cells expressing a tagged version of PanCoVac, we confirmed in vitro that PanCoVac is processed and presented by HLA-A*0201. As mucosal immunity in the respiratory tract is crucial for protection against respiratory viruses such as SARS-CoV-2, we tested the protective effect of single-low dose of NILV-PanCoVac administered via the intranasal (i.n.) route in the Roborovski dwarf hamster model of COVID-19. After infection with ancestral SARS-CoV-2, animals immunized with a single-low dose of NILV-PanCoVac i.n. did not show symptoms and had significantly decreased viral loads in the lung tissue. This protective effect was observed in the early phase (2 days post infection) after challenge and was not dependent on neutralizing antibodies. PanCoVac, a multi-epitope vaccine covering conserved T cell epitopes from all structural proteins of coronaviruses, might protect from severe disease caused by SARS-CoV-2 variants and future pathogenic coronaviruses. The use of (HLA-) humanized animal models will allow for further efficacy studies of PanCoVac-based vaccines in vivo.


Impact of Tooth Loss Position on Oral Health-Related Quality of Life in Adults Treated in the Community.
Tooth loss is known to have negative effects on both functional and psychological oral health-related quality of life (OHRQoL), but the impact of the position of the tooth loss (i.e. anterior or posterior) on the different psychosocial dimensions of OHRQoL has yet to be examined. Here, we examined how the position of lost teeth impacts the different dimensions of OHRQoL. This was a cross-sectional epidemiological study of adults aged 18 years and older attending routine examinations at primary care dental centers in Jeddah, Kingdom of Saudi Arabia. Demographic information was collected, and OHRQoL was assessed using the Oral Health Impact Profile-14 (OHIP-14) (Arabic form). Differences in total and subdomain OHIP-14 scores between individuals without tooth loss and those with ≥1 anterior or posterior missing teeth were assessed using Student's t-test, and analysis of covariance was used to assess the association between the presence and absence of missing teeth in each compartment and total and subdomain OHIP-14 scores after controlling for age, gender, and income. The overall prevalence of tooth loss was 76%. In multivariate analysis controlling for age, gender, and income as covariates, anterior missing teeth were significantly associated with higher OHIP-14 total, physical pain, physical disability, psychological disability, and social disability scores, accounting for 6%-12% of the score variance. However, posterior missing teeth were only associated with total OHIP-14 and functional limitations domain scores, accounting for 6% and 7% of the variance, respectively. Here, we show for the first time the impact of the location of missing teeth on different OHRQoL dimensions. Anterior tooth loss has a wide-ranging impact on both physical and psychosocial functioning compared to posterior tooth loss, suggesting that anterior tooth restoration should be prioritized when treatment planning. The position of lost teeth must be considered in addition to the number of losses when examining the impact of tooth loss and its treatment on OHRQoL.


Emergence and epidemiology of ciguatera in the coastal cities of southern China.
In the present review of 23 published case studies, the main objective is to report the emergence and epidemiology of ciguatera in the coastal cities of southern China. There was a sudden surge in ciguatera outbreaks in 2004. Ciguatera mostly occurred in the Guangdong Province. In Shenzhen, the incidence of ciguatera in 2004 was estimated to be over 7.5 per million people. In Foshan and Zhongshan, three large outbreaks each affecting over 100-200 subjects (caused by tiger grouper served at banquets) accounted for the much higher incidence of ciguatera in 2004 (>48.7 and >129.9 per million people). Humphead wrasse and areolated coral grouper were the other important ciguatoxic fish. In some subjects, risk factors for increased likelihood of (severe) ciguatera were present, namely concomitant alcohol consumption and ingestion of large reef fishes and CTX-rich fish parts. To prevent large outbreaks and severe illness, large apex predators from coral reefs should never be served at banquets and the public should realize the increased risk of severe symptoms due to ingestion of CTX-rich fish parts with alcohol. The systematic collection of accurate details, implementation of risk assessment process and continuing education for the public on prevention are of obvious importance.


Enzyme-linked immunosorbent assays adapted for serotyping of human rotavirus strains.
Five enzyme-linked immunosorbent assay systems were adapted for serotyping human rotavirus strains and were compared with a sensitive complement fixation test in terms of specificity and sensitivity. The assays differed mainly with regard to the antibody systems involved in the double sandwich. Serotype differentiation of 34 rotavirus strains was achieved by determining a neutralization endpoint titer, either with a constant antiserum-varying antigen dilution method or vice versa. The procedure which proved to be highly specific and sensitive was one with two type-specific hyperimmune sera (enzyme-linked immunosorbent assay system 5) instead of only one, as in the four other systems.


[Comparison of in vivo characteristics of polyethylene wear particles produced by a metal and a ceramic femoral component in total knee replacement].
The aim of the study was to evaluate in vivo and compare, in terms of the quality and number of ultra high-molecular polyethylene (UHMWPE) wear particles, total knee replacements of identical construction differing only in the material used for femoral component production, i.e., CoCrMo alloy or ZrO2 ceramics. Samples of peri-prosthetic granuloma tissue were collected in two patients with total knee replacement suffering from implant migration, who were matched in relevant characteristics. The primary knee replacement in Patient 1 with a CoCrMo femoral component was done 7.2 years and in Patient 2 with a ZrO2 implant 6.8 years before this assessment. The polyethylene wear-induced granuloma was analysed by the MORF method enabling us to assess the shape and size of wear debris and the IRc method for assessment of particle concentration. In the granuloma tissue samples of Patient 1, on the average, particles were 0.30 mm in size and their relative volume was 0.19. In the Patient 2 tissue samples, the average size of particles was 0.33 mm and their relative volume was 0.26. There was no significant difference in either particle morphology or their concentration in the granuloma tissue between the two patients. One of the options of how to reduce the production of polyethylene wear particles is to improve the tribological properties of contacting surfaces in total knee replacement by substituting a cobalt-chrome femoral component with a zirconia ceramic femoral component. The previous in vitro testing carried out with a mechanical simulator under conditions approaching real weight-bearing in the human body did show a nearly three-fold decrease in the number of UHMWPE wear particles in zirconia components. The evaluation of granuloma tissue induced by the activity of a real prosthetic joint for nearly seven years, however, did not reveal any great difference in either quality or quantity of polyethylene debris between the two replacements. The difference of surface roughness between CoCrMo (Ra = 0.05) and ZrO2 (Ra = 0.02) components did not play any role in in vivo conditions. CONCLUSIONS In accordance with a previous clinical study, this evaluation of the quality and quantity of UHMWPE wear particles produced by a ceramic femoral component in vivo failed to demonstrate any advantage of zirconia ceramic components over the cobalt-chrome femoral components so far used.


Stimulation by unsaturated fatty acid of squalene uptake in rat liver microsomes.
Supernatant protein factor (SPF) and anionic phospholipids such as phosphatidylglycerol (PG) stimulate squalene epoxidase activity in rat liver microsomes by promoting [3H]squalene uptake as well as substrate translocation (Chin, J., and K. Bloch. 1984. J. Biol. Chem. 259: 11735-11738). This process is postulated to be membrane-mediated and not carrier-mediated. Here we show that treatment of PG with phospholipase A2 in the presence of bovine serum albumin abolishes the stimulatory effect of SPF on epoxidase activity. Disaturated fatty acyl-PGs are not as effective as egg yolk lecithin PG in the SPF effect. These findings suggest an important role for the unsaturated fatty acid moiety of PG. We also show that at submicellar concentrations, cis-unsaturated fatty acids stimulate microsomal epoxidase activity whereas saturated fatty acids do not. This effect is due to an increase in substrate uptake which in turn may facilitate substrate availability to the enzyme.


Activation of peroxisome proliferator-activated receptor-{delta} by GW501516 prevents fatty acid-induced nuclear factor-{kappa}B activation and insulin resistance in skeletal muscle cells.
Elevated plasma free fatty acids cause insulin resistance in skeletal muscle through the activation of a chronic inflammatory process. This process involves nuclear factor (NF)-kappaB activation as a result of diacylglycerol (DAG) accumulation and subsequent protein kinase Ctheta (PKCtheta) phosphorylation. At present, it is unknown whether peroxisome proliferator-activated receptor-delta (PPARdelta) activation prevents fatty acid-induced inflammation and insulin resistance in skeletal muscle cells. In C2C12 skeletal muscle cells, the PPARdelta agonist GW501516 prevented phosphorylation of insulin receptor substrate-1 at Ser(307) and the inhibition of insulin-stimulated Akt phosphorylation caused by exposure to the saturated fatty acid palmitate. This latter effect was reversed by the PPARdelta antagonist GSK0660. Treatment with the PPARdelta agonist enhanced the expression of two well known PPARdelta target genes involved in fatty acid oxidation, carnitine palmitoyltransferase-1 and pyruvate dehydrogenase kinase 4 and increased the phosphorylation of AMP-activated protein kinase, preventing the reduction in fatty acid oxidation caused by palmitate exposure. In agreement with these changes, GW501516 treatment reversed the increase in DAG and PKCtheta activation caused by palmitate. These effects were abolished in the presence of the carnitine palmitoyltransferase-1 inhibitor etomoxir, thereby indicating that increased fatty acid oxidation was involved in the changes observed. Consistent with these findings, PPARdelta activation by GW501516 blocked palmitate-induced NF-kappaB DNA-binding activity. Likewise, drug treatment inhibited the increase in IL-6 expression caused by palmitate in C2C12 and human skeletal muscle cells as well as the protein secretion of this cytokine. These findings indicate that PPARdelta attenuates fatty acid-induced NF-kappaB activation and the subsequent development of insulin resistance in skeletal muscle cells by reducing DAG accumulation. Our results point to PPARdelta activation as a pharmacological target to prevent insulin resistance.


Igf signaling couples retina growth with body growth by modulating progenitor cell division.
How the body and organs balance their relative growth is of key importance for coordinating size and function. This is of particular relevance in organisms, which continue to grow over their entire life span. We addressed this issue in the neuroretina of medaka fish (Oryzias latipes), a well-studied system with which to address vertebrate organ growth. We reveal that a central growth regulator, Igf1 receptor (Igf1r), is necessary and sufficient for proliferation control in the postembryonic retinal stem cell niche: the ciliary marginal zone (CMZ). Targeted activation of Igf1r signaling in the CMZ uncouples neuroretina growth from body size control, and we demonstrate that Igf1r operates on progenitor cells, stimulating their proliferation. Activation of Igf1r signaling increases retinal size while preserving its structural integrity, revealing a modular organization in which progenitor differentiation and neurogenesis are self-organized and highly regulated. Our findings position Igf signaling as a key module for controlling retinal size and composition, with important evolutionary implications.


A review of evolution of electrospun tissue engineering scaffold: From two dimensions to three dimensions.
The potential of electrospinning process to fabricate ultrafine fibers as building blocks for tissue engineering scaffolds is well recognized. The scaffold construct produced by electrospinning process depends on the quality of the fibers. In electrospinning, material selection and parameter setting are among many factors that contribute to the quality of the ultrafine fibers, which eventually determine the performance of the tissue engineering scaffolds. The major challenge of conventional electrospun scaffolds is the nature of electrospinning process which can only produce two-dimensional electrospun mats, hence limiting their applications. Researchers have started to focus on overcoming this limitation by combining electrospinning with other techniques to fabricate three-dimensional scaffold constructs. This article reviews various polymeric materials and their composites/blends that have been successfully electrospun for tissue engineering scaffolds, their mechanical properties, and the various parameters settings that influence the fiber morphology. This review also highlights the secondary processes to electrospinning that have been used to develop three-dimensional tissue engineering scaffolds as well as the steps undertaken to overcome electrospinning limitations.


Isolation and characterization of PSI-LHCI super-complex and their sub-complexes from a red alga Cyanidioschyzon merolae.
Photosystem I (PSI)-light-harvesting complex I (LHCI) super-complex and its sub-complexes PSI core and LHCI, were purified from a unicellular red alga Cyanidioschyzon merolae and characterized. PSI-LHCI of C. merolae existed as a monomer with a molecular mass of 580 kDa. Mass spectrometry analysis identified 11 subunits (PsaA, B, C, D, E, F, I, J, K, L, O) in the core complex and three LHCI subunits, CMQ142C, CMN234C, and CMN235C in LHCI, indicating that at least three Lhcr subunits associate with the red algal PSI core. PsaG was not found in the red algae PSI-LHCI, and we suggest that the position corresponding to Lhca1 in higher plant PSI-LHCI is empty in the red algal PSI-LHCI. The PSI-LHCI complex was separated into two bands on native PAGE, suggesting that two different complexes may be present with slightly different protein compositions probably with respective to the numbers of Lhcr subunits. Based on the results obtained, a structural model was proposed for the red algal PSI-LHCI. Furthermore, pigment analysis revealed that the C. merolae PSI-LHCI contained a large amount of zeaxanthin, which is mainly associated with the LHCI complex whereas little zeaxanthin was found in the PSI core. This indicates a unique feature of the carotenoid composition of the Lhcr proteins and may suggest an important role of Zea in the light-harvesting and photoprotection of the red algal PSI-LHCI complex.


Na+/K+‑ATPase DR region‑specific antibody protects U251 cells against hypoxia reperfusion‑induced injury via the PI3K/AKT and ERK pathways.
Cerebral ischemia is a condition in which there is insufficient blood flow to the brain to meet metabolic demand. This leads to poor oxygen supply or cerebral hypoxia and to the death of brain tissue or cerebral infarction/ischemic stroke. In the present study, an Na+/K+‑ATPase (NKA) DR region‑specific antibody (DRSAb) was established and purified and it was demonstrated that DRSAb induced a protective effect on human astrocytes (U251) via the phosphoinositide 3‑kinase (PI3K)/AKT and extracellular signal‑regulated protein kinase (ERK) signaling pathways. The binding of DRSAb on NKA was revealed using flow cytometry. High signals were detected on U251 cells incubated with DRSAb, but not with control sera or BSA. The viability of the hypoxia/reperfusion (H/R)‑treated cells was markedly increased by DRSAb administration of 0.3‑0.5 µM. The optimal concentration of DRSAb was 0.4 µM for attenuation of the injury induced by H/R. The administration of 0.4 µM DRSAb markedly reduced the number of apoptotic cells compared with control sera. The application of PD98059, an ERK inhibitor, and LY‑294002, an AKT inhibitor, attenuated the protective effect induced by DRSAb in the U251 cells subjected to H/R. Furthermore, the application of LY294002 prior to incubation with DRSAb eliminated the activation of ERK1/2, whereas the use of PD98059 failed to attenuate the effect of DRSAb on PI3K/AKT activation. These results indicated that the protective effects of DRSAb against H/R injury in U251 cells occurred via stimulation of the PI3K/AKT and ERK signaling pathways.


Development of a structured generic drug intervention model for public health purposes: a brief application of motivational interviewing with young people.
Brief applications of Motivational Interviewing (MI) emerged around 15 years ago to target problematic alcohol and other drug use. Interventions which specifically target illicit drug use, young people, or which are delivered in settings other than health-care services have, however, been relatively slow to develop. The needs of young people for interventions distinct from those offered to adults are considered, as a precursor to an outline of the structure of a newly adapted intervention targeting drug use in general among young people. Based upon earlier topic-based approaches developed by Rollnick et al. this intervention is innovative in simultaneously targeting a range of drugs in pursuit of secondary prevention objectives, while also seeking to manifest the spirit of MI. The intervention consists of a single-session face-to-face conversation of up to 60 minutes duration. Data are presented which describe the development and conduct of this intervention during the course of an efficacy trial, with promising efficacy data themselves reported elsewhere. Observations are made on intervention delivery and consideration is given to implications for further novel targeting of young people and within the field of addiction interventions more generally. [McCambridge J, Strang J. Development of a structured generic drug intervention model for public health purposes: a brief application of motivational interviewing with young people.


Founding of the Association of Physician Assistant Programs and the Organization's Central Role in the Development of the Physician Assistant Profession.
The Physician Assistant Education Association (PAEA) (formerly Association of Physician Assistant Programs [APAP]) was founded in 1972 by early PA program leaders to encourage collaboration and discussion among program leaders and faculty on a wide range of issues of mutual concern. This article addresses the founding of the organization, which continues to represent PA programs today. It addresses the important accomplishments of APAP during the 1972-1974 period and describes in detail the essential contributions of APAP and its leaders in developing the PA profession. Included are discussions of "the Role of the Registry of Physicians' Associates," which was incorporated into APAP; the sponsorship of "the First National Conference on New Health Practitioners" in collaboration with AAPA; the "Launching a Joint National Office" for APAP and AAPA in Washington, DC, in 1973; and "Places at the Table," which reviews the successful efforts of APAP leaders to gain inclusion of AAPA and APAP in the Joint Review Committee on Educational Programs for Assistants to the Primary Physician (JRC-PA) for accrediting qualified programs, the contribution of APAP leaders to the development of the first Certification Examination for the Assistant to the Primary Care Physician by the National Board of Medical Examiners (NBME) in 1973, leadership in the founding of the National Commission on the Certification of Physician Assistants (NCCPA) in 1974, and collaboration with the Association of American Medical Colleges (AAMC) in developing interdisciplinary education and training. It concludes with a summation of the legacy of APAP (PAEA's) formative years.


[Morphological changes of lymphoid apparatus of the larynx after experimental exposure to various balneal factors].
The aim of this investigation was to detect the structural changes of lymphoid components of rat pharynx in an experiment after a course of exposure to various balneal procedures. The studies were performed on 90 outbred mature 3 month-old male rats (20 animals in each experimental group and 10 animals in each control groups). The animals were exposed to a course of weakly mineralized organic bituminous, thermal iodobromine and strong (concentrated) sulfide baths present on the territory of Azerbaijan. The experiments performed have shown a significant sensitivity of the lymphoid structures of the rat pharynx to the balneal procedures. After exposure to iodobromine and bituminous baths, the signs of lymphocytopoiesis activation were noted. The exposure to strong sulfide baths resulted in a morphological regression of lymphoid apparatus of rat larynx, which raises the question on the expediency of the use of these procedures in practical balneology.


Data pooling and analysis to build a preliminary item bank: an example using bowel function in prostate cancer.
Assessing bowel function (BF) in prostate cancer can help determine therapeutic trade-offs. We determined the components of BF commonly assessed in prostate cancer studies as an initial step in creating an item bank for clinical and research application. We analyzed six archived data sets representing 4,246 men with prostate cancer. Thirty-one items from validated instruments were available for analysis. Items were classified into domains (diarrhea, rectal urgency, pain, bleeding, bother/distress, and other) then subjected to conventional psychometric and item response theory (IRT) analyses. Items fit the IRT model if the ratio between observed and expected item variance was between 0.60 and 1.40. Four of 31 items had inadequate fit in at least one analysis. Poorly fitting items included bleeding (2), rectal urgency (1), and bother/distress (1). A fifth item assessing hemorrhoids was poorly correlated with other items. Our analyses supported four related components of BF: diarrhea, rectal urgency, pain, and bother/distress.


Effect of an electric field on an intermittent granular flow.
Granular gravity driven flows of glass beads have been observed in a silo with a flat bottom. A dc high electric field has been applied perpendicularly to the silo to tune the cohesion. The outlet mass flow has been measured. An image subtraction technique has been applied to visualize the flow geometry and a spatiotemporal analysis of the flow dynamics has been performed. The outlet mass flow is independent of voltage, but a transition from funnel flow to rathole flow is observed. This transition is of probabilistic nature and an intermediate situation exists between the funnel and the rathole situations. At a given voltage, two kinds of flow dynamics can occur: a continuous flow or an intermittent flow. The electric field increases the probability to observe an intermittent flow.


Warburg effect and translocation-induced genomic instability: two yeast models for cancer cells.
Yeast has been established as an efficient model system to study biological principles underpinning human health. In this review we focus on yeast models covering two aspects of cancer formation and progression (i) the activity of pyruvate kinase (PK), which recapitulates metabolic features of cancer cells, including the Warburg effect, and (ii) chromosome bridge-induced translocation (BIT) mimiking genome instability in cancer. Saccharomyces cerevisiae is an excellent model to study cancer cell metabolism, as exponentially growing yeast cells exhibit many metabolic similarities with rapidly proliferating cancer cells. The metabolic reconfiguration includes an increase in glucose uptake and fermentation, at the expense of respiration and oxidative phosphorylation (the Warburg effect), and involves a broad reconfiguration of nucleotide and amino acid metabolism. Both in yeast and humans, the regulation of this process seems to have a central player, PK, which is up-regulated in cancer, and to occur mostly on a post-transcriptional and post-translational basis. Furthermore, BIT allows to generate selectable translocation-derived recombinants ("translocants"), between any two desired chromosomal locations, in wild-type yeast strains transformed with a linear DNA cassette carrying a selectable marker flanked by two DNA sequences homologous to different chromosomes. Using the BIT system, targeted non-reciprocal translocations in mitosis are easily inducible. An extensive collection of different yeast translocants exhibiting genome instability and aberrant phenotypes similar to cancer cells has been produced and subjected to analysis. In this review, we hence provide an overview upon two yeast cancer models, and extrapolate general principles for mimicking human disease mechanisms in yeast.


Assembly and function of type III secretory systems.
Type III secretion systems allow Yersinia spp., Salmonella spp., Shigella spp., Bordetella spp., and Pseudomonas aeruginosa and enteropathogenic Escherichia coli adhering at the surface of a eukaryotic cell to inject bacterial proteins across the two bacterial membranes and the eukaryotic cell membrane to destroy or subvert the target cell. These systems consist of a secretion apparatus, made of approximately 25 proteins, and an array of proteins released by this apparatus. Some of these released proteins are "effectors," which are delivered into the cytosol of the target cell, whereas the others are "translocators," which help the effectors to cross the membrane of the eukaryotic cell. Most of the effectors act on the cytoskeleton or on intracellular-signaling cascades. A protein injected by the enteropathogenic E. coli serves as a membrane receptor for the docking of the bacterium itself at the surface of the cell. Type III secretion systems also occur in plant pathogens where they are involved both in causing disease in susceptible hosts and in eliciting the so-called hypersensitive response in resistant or nonhost plants. They consist of 15-20 Hrp proteins building a secretion apparatus and two groups of effectors: harpins and avirulence proteins. Harpins are presumably secreted in the extracellular compartment, whereas avirulence proteins are thought to be targeted into plant cells. Although a coherent picture is clearly emerging, basic questions remain to be answered. In particular, little is known about how the type III apparatus fits together to deliver proteins in animal cells. It is even more mysterious for plant cells where a thick wall has to be crossed. In spite of these haunting questions, type III secretion appears as a fascinating trans-kingdom communication device.


Delayed contrast enhancement and perfusable tissue index in hypertrophic cardiomyopathy: comparison between cardiac MRI and PET.
Delayed contrast enhancement (DCE) visualized by cardiac MRI (CMR) is a common feature in patients with hypertrophic cardiomyopathy (HCM), presumed to be related to myocardial fibrosis. The pathophysiologic basis of hyperenhancement in this patient group, however, remains unclear as limited histologic comparisons are available. The present study compares the perfusable tissue index (PTI), an alternative marker of myocardial fibrosis obtained by PET, with DCE-CMR in HCM. Twenty-one patients with asymmetric septal HCM, 12 chronic myocardial infarction (MI) patients, and 6 age-matched healthy control subjects were studied with DCE-CMR and PET. PET was performed using (15)O-labeled water and carbon monoxide to obtain the PTI. No hyperenhancement was observed in control subjects and the PTI was within normal limits (1.10 +/- 0.07 [mean +/- SD]). In MI patients, the extent of hyperenhancement (25% +/- 16% [mean +/- SD]) was inversely related to the decrease in the PTI (0.94 +/- 0.12; r = -0.65, P < 0.05). Average hyperenhancement in HCM was 14% +/- 12%, predominantly located in the interventricular septum. The PTI in the hypertrophied interventricular septum, however, was not reduced (1.12 +/- 0.13). Furthermore, in contrast to MI patients, there was a modest positive correlation between the extent of DCE and the PTI in HCM (r = 0.45, P < 0.05). DCE in the hypertrophied septum of HCM patients is not accompanied by a decline in the PTI, and there is a positive correlation between the extent of DCE and the PTI. These results suggest that hyperenhancement may not be caused solely by fibrotic replacement scarring in this patient group. Other pathologic changes associated with HCM may also cause gadolinium-diethylenetriaminepentaacetic acid hyperenhancement.


A biopsychosocial model of glycemic control in diabetes: stress, coping and regimen adherence.
This study examines stress, coping, and regimen adherence as determinants of chronic and transient metabolic control in diabetes. We also examine the interaction of biologic vulnerability and psychosocial risk factors to see if Type 1 (insulin dependent diabetes mellitus) or Type 2 (noninsulin dependent diabetes mellitus) diabetes had greater responsiveness to psychosocial risk factors. Analyses of data from insulin-treated adults with Type 1 (N = 57) and Type 2 (N = 61) diabetes supported the biopsychosocial model. For Type 1 diabetes, self-controlling persons had better glycemic control and emotional persons had worse (because of differences in stress). All of these associations were mediated by regimen compliance. For Type 2 diabetes, self-controlling persons had better glycemic control for reasons other than regimen compliance. There was an interaction between biologic and psychosocial factors, with psychosocial factors accounting for more variance in glycemic control within Type 1 patients. Stable psychosocial resources (i.e., education, being married, and positive coping styles) were associated with better chronic glycemic control, while stress and regimen nonadherence were associated with worse transient glycemic control.


Intravitreal triamcinolone for the treatment of macular edema associated with central retinal vein occlusion.
To investigate the safety and efficacy of intravitreal triamcinolone acetonide as treatment for macular edema associated with central retinal vein occlusion (CRVO). We reviewed the medical records of 13 consecutive patients (13 eyes) with macular edema associated with CRVO who were treated with an injection of intravitreal triamcinolone acetonide (4 mg) at the University of Wisconsin and the Bascom Palmer Eye Institute. Each intravitreal injection was delivered through the pars plana using a 27- or 30-gauge needle. Change in Snellen visual acuity, clinical appearance of macular edema, measurement of foveal thickening with optical coherence tomography (OCT), and frequency of complications. The median age of the 13 patients was 67 years (interquartile range, 57-77 years), and the median duration of symptoms before injection was 8 months (interquartile range, 4-9 months). Mean baseline visual acuity was 20/500 in the affected eye. Mean visual acuity at the 6-month follow-up examination was 20/180 in the affected eye. All 13 patients completed the 6-month examination. Eyes with nonischemic CRVO (n = 5) demonstrated a significant improvement in visual acuity, whereas eyes with ischemic CRVO (n = 8) demonstrated a nonsignificant visual acuity improvement. No patient had a decrease in visual acuity. Mean baseline foveal thickness as measured by OCT was 590 micro m (retinal thickening = 416 micro m). Mean foveal thickness as measured by OCT at the 1-month follow-up examination in 12 patients was 212 micro m (retinal thickening = 38 micro m). At the 3-month follow-up examination, mean foveal thickness as measured by OCT for 13 patients was 193 micro m (retinal thickening = 19 micro m). Between the 3- and 6-month follow-up examinations, 4 patients developed a recurrence of macular edema. Three of the 4 patients were retreated with a second injection of triamcinolone. Two of these 3 patients experienced an improvement in visual acuity following retreatment. At the 6-month follow-up examination, mean foveal thickness as measured by OCT for 13 patients was 281 micro m (retinal thickening = 107 micro m). No adverse effects such as retinal detachment or endophthalmitis occurred. One patient experienced an increase in intraocular pressure that was controlled with 2 aqueous suppressants. Intravitreal injection of triamcinolone appears to be a possibly effective treatment in some patients with macular edema associated with CRVO. Patients with nonischemic CRVO may respond more favorably than patients with ischemic CRVO, and retreatment may be necessary in some patients. In this case series, severe complications were not noted.


Noninvasive functional optical spectroscopy of human breast tissue.
Near infrared diffuse optical spectroscopy and diffuse optical imaging are promising methods that eventually may enhance or replace existing technologies for breast cancer screening and diagnosis. These techniques are based on highly sensitive, quantitative measurements of optical and functional contrast between healthy and diseased tissue. In this study, we examine whether changes in breast physiology caused by exogenous hormones, aging, and fluctuations during the menstrual cycle result in significant alterations in breast tissue optical contrast. A noninvasive quantitative diffuse optical spectroscopy technique, frequency-domain photon migration, was used. Measurements were performed on 14 volunteer subjects by using a hand-held probe. Intrinsic tissue absorption and reduced scattering parameters were calculated from frequency-domain photon migration data. Wavelength-dependent absorption (at 674, 803, 849, and 956 nm) was used to determine tissue concentration of oxyhemoglobin, deoxyhemoglobin, total hemoglobin, tissue hemoglobin oxygen saturation, and bulk water content. Results show significant and dramatic differences in optical properties between menopausal states. Average premenopausal intrinsic tissue absorption and reduced scattering values at each wavelength are 2.5- to 3-fold higher and 16-28 % greater, respectively, than absorption and scattering for postmenopausal subjects. Absorption and scattering properties for women using hormone replacement therapy are intermediate between premenopausal and postmenopausal populations. Physiological properties show differences in mean total hemoglobin (7.0 microM, 11.8 microM, and 19.2 microM) and water concentration relative to pure water (10.9 %, 15.3 %, and 27.3 %) for postmenopausal, hormone replacement therapy, and premenopausal subjects, respectively. Because of their unique, quantitative information content, diffuse optical methods may play an important role in breast diagnostics and improving our understanding of breast disease.


Use of a monoclonal antibody to detect elevated levels of a modified nucleoside, N-[9-(beta-D-ribofuranosyl)purin-6-ylcarbamoyl]-L-threonine, in the urine of breast cancer patients.
Monoclonal antibodies to the modified nucleoside N-[9-(beta-D-ribofuranosyl)purin-6-ylcarbamoyl]-L-threonine (t6A) have been produced and characterized. These antibodies were utilized in a radioimmunoassay to quantitate the levels of this modified nucleoside in the urine of patients with benign breast disease, primary breast cancer, and metastatic breast cancer. Eighty-nine % (16 of 18) of patients with metastatic breast cancer excreted higher quantities of t6A than did patients with benign breast disease or primary breast cancer. This modified nucleoside represents a new, potential marker not previously included in similar studies using high-performance liquid chromatography. The immunoassay approach for quantitating this nucleoside has the advantage over high-performance liquid chromatography in that the former is more compatible with a clinical laboratory setting, and it does not require pretreatment of the urine or sophisticated analytical equipment. The elevation of t6A levels in metastatic breast cancer patients indicates that it may be a more sensitive marker than previously studied modified nucleosides. Furthermore, t6A might be particularly useful for monitoring transition to the metastatic stage in patients already diagnosed as having breast cancer.


Serum immunoglobulin E (Ig E) levels after myocardial infarction.
The relation between cardiovascular diseases and serum immunoglobulin E (Ig E) levels has been investigated by different authors. Serum Ig E may play a direct role in the pathogenesis of cardiovascular diseases, or it may only be a marker formed during pathological mechanisms. In this study, we determined serum Ig E levels of patients with unstable angina pectoris and myocardial infarction. Serum Ig E levels of 30 patients with acute myocardial infarction and 20 patients with unstable angina pectoris were determined within the first 6 and 12th hours and on 2nd, 4th, 8th days of the hospitalization and these levels were compared with the serum Ig E levels of normal subjects reserved as control group. As a result, at all measurement hours, serum Ig E levels of acute myocardial infarction group were found to be approximately fourfold increased when compared to those of unstable angina pectoris and those of control group (p < 0.001). In the acute myocardial infarction group, no significant differences could be found between the levels of the first 6th and 12th hours and on the 2nd, 4th, 8th days. Also, no statistically significant difference was found between the serum Ig E levels of unstable angina pectoris and those of control group.


The role of neuronal synchronization in selective attention.
Attention selectively enhances the influence of neuronal responses conveying information about relevant sensory attributes. Accumulating evidence suggests that this selective neuronal modulation relies on rhythmic synchronization at local and long-range spatial scales: attention selectively synchronizes the rhythmic responses of those neurons that are tuned to the spatial and featural attributes of the attended sensory input. The strength of synchronization is thereby functionally related to perceptual accuracy and behavioural efficiency. Complementing this synchronization at a local level, attention has recently been demonstrated to regulate which locally synchronized neuronal groups phase-synchronize their rhythmic activity across long-range connections. These results point to a general computational role for selective synchronization in dynamically controlling which neurons communicate information about sensory inputs effectively.


Early adaptive developments of Pseudomonas aeruginosa after the transition from life in the environment to persistent colonization in the airways of human cystic fibrosis hosts.
Pseudomonas aeruginosa is an opportunistic pathogen ubiquitous to the natural environment but with the capability of moving to the host environment. Long-term infection of the airways of cystic fibrosis patients is associated with extensive genetic adaptation of P. aeruginosa, and we have studied cases of the initial stages of infection in order to characterize the early adaptive processes in the colonizing bacteria. A combination of global gene expression analysis and phenotypic characterization of longitudinal isolates from cystic fibrosis patients revealed well-known characteristics such as conversion to a mucoid phenotype by mucA mutation and increased antibiotic resistance by nfxB mutation. Additionally, upregulation of the atu operon leading to enhanced growth on leucine provides a possible example of metabolic optimization. A detailed investigation of the mucoid phenotype uncovered profound pleiotropic effects on gene expression including reduction of virulence factors and the Rhl quorum sensing system. Accordingly, mucoid isolates displayed a general reduction of virulence in the Caenorhabditis elegans infection model, altogether suggesting that the adaptive success of the mucoid variant extends beyond the benefits of alginate overproduction. In the overall perspective the global phenotype of the adapted variants appears to place them on paths in direction of fully adapted strains residing in long-term chronically infected patients.


Fourier transform infrared spectroscopic studies of the interaction of the antimicrobial peptide gramicidin S with lipid micelles and with lipid monolayer and bilayer membranes.
We have utilized Fourier transform infrared spectroscopy to study the interaction of the antimicrobial peptide gramicidin S (GS) with lipid micelles and with lipid monolayer and bilayer membranes as a function of temperature and of the phase state of the lipid. Since the conformation of GS does not change under the experimental conditions employed in this study, we could utilize the dependence of the frequency of the amide I band of the central beta-sheet region of this peptide on the polarity and hydrogen-bonding potential of its environment to probe GS interaction with and location in these lipid model membrane systems. We find that the GS is completely or partially excluded from the gel states of all of the lipid bilayers examined in this study but strongly partitions into lipid micelles, monolayers, or bilayers in the liquid-crystalline state. Moreover, in general, the penetration of GS into zwitterionic and uncharged lipid bilayer coincides closely with the gel to liquid-crystalline phase transition of the lipid. However, GS begins to penetrate into the gel-state bilayers of anionic phospholipids prior to the actual chain-melting phase transition, while in cationic lipid bilayers, GS does not partition strongly into the liquid-crystalline bilayer until temperatures well above the chain-melting phase transition are reached. In the liquid-crystalline state, the polarity of the environment of GS indicates that this peptide is located primarily at the polar/apolar interfacial region of the bilayer near the glycerol backbone region of the lipid molecule. However, the depth of GS penetration into this interfacial region can vary somewhat depending on the structure and charge of the lipid molecule. In general, GS associates most strongly with and penetrates most deeply into more disordered bilayers with a negative surface charge, although the detailed chemical structure of the lipid molecule and physical organization of the lipid aggregate (micelle versus monolayer versus bilayer) also have minor effects on these processes.


Providers caring for adolescents with perinatally-acquired HIV: Current practices and barriers to communication about sexual and reproductive health.
The population of adolescents and young adults (AYA) with perinatally-acquired HIV (PHIV) present challenges to HIV healthcare providers (HHCPs). Originally not expected to survive childhood, they are now living well into young adulthood. Little is known about the type of sexual and reproductive (SRH) information/services offered to AYA with PHIV by HHCPs. HHCPs (n=67) were recruited using snowball sampling, and completed an online survey. Providers' most frequently endorsed SRH topics discussed with both male and female patients included condom use (77.3%), STD prevention (73.1%), and screening (62.1%). Providers' reports indicated that females received significantly more education about SRH topics overall. The most frequently noted barriers to SRH communication included more pressing health concerns (53.0%), parent/guardian not receptive (43.9%), and lack of time during appointment (43.9%). Provider-reported SRH conversations with HHCPs were highly focused on horizontal transmission and pregnancy prevention. Salient social aspects of SRH promotion for AYAs with PHIV (e.g., managing disclosure and romantic relationships) were less commonly discussed, though such conversations may serve to reduce secondary transmission and enhance the overall well-being of AYA with PHIV. Findings indicated that further work must be done to identify strategies to address unmet SRH needs of the aging population of AYA with PHIV.


Human adrenocortical cell xenotransplantation: model of cotransplantation of human adrenocortical cells and 3T3 cells in scid mice to form vascularized functional tissue and prevent adrenal insufficiency.
To establish an experimental model for replacement of endocrine organ function by xenotransplantation, human adrenocortical cells from postnatal donors were transplanted beneath the kidney capsule of adrenalectomized scidmice together with mitomycin C-treated 3T3 cells that secrete FGF. Adrenocortical cells from seven donors, male and female, ranging from 6-50 years of age, were used. 12 of 13 animals survived > 16 days following surgery. After 50 days they were sacrified to allow assessment of the histology and ultrastructure of tissue formed from the transplanted cells. Only 1 of 23 adrenalectomized sham-operated animals survived > 16 days. In all surviving animals, vascularized adrenocortical tissue formed at the site of transplantation. Cortisol, the normal human glucocorticoid, was present in the plasma of these animals, replacing corticosterone, the mouse glucocorticoid. Some animals, but not most, had measurable aldosterone. The tissue formed from the transplanted cells showed histological and ultrastructural features of normal adrenal cortex. Mitochondria had tubulo-vesicular cristae and there were prominent microvilli between cells. Tissues had a well-developed vasculature, sometimes with large sinusoidal vessels. Proliferation in the transplant tissues was very low. These results show that tissue formed from transplanted human adrenocortical cells is able to replace the essential functions of the adrenal gland in scid mice. This demonstrates that transplanted human endocrine cells can functionally replace a surgically removed endocrine organ in a host animal.


Variation in Length of Stay by Level of Neonatal Care Among Moderate and Late Preterm Infants.
Moderate and late preterm infants are a growing subgroup of neonates with increased care needs after birth, yet standard protocols are lacking. We aim to describe variation in length of stay (LOS) by gestational age (GA) across hospitals within the same level of neonatal care and between different levels of neonatal care. Retrospective cohort study of hospitalizations for moderate (32-33 weeks GA) and late (34-36 weeks GA) preterm infants in 2019 Kid's Inpatient Database. We compared adjusted LOS in this cohort and evaluated variation within hospitals of the same level and across different levels of neonatal care. This study includes 217 051 moderate (26.2%) and late (73.8%) preterm infants from level II (19.7%), III (66.3%), and IV (11.1%) hospitals. Patient-level (race and ethnicity, primary payor, delivery type, multiple gestation, birth weight) and hospital-level (birth region, level of neonatal care) factors were significantly associated with LOS. Adjusted mean LOS varied for hospitals within the same level of neonatal care with level II hospitals showing the greatest variability among 34- to 36- week GA infants when compared with level III and IV hospitals (P < .01). LOS also varied significantly between levels of neonatal care with the greatest variation (0.9 days) seen in 32-week GA between level III and level IV hospitals. For moderate and late preterm infants, the level of neonatal care was associated with variation in LOS after adjusting for clinical severity. Hospitals providing level II neonatal care showed the greatest variation and may provide an opportunity to standardize care.


Shining Damaged Hearts: Immunotherapy-Related Cardiotoxicity in the Spotlight of Nuclear Cardiology.
The emerging use of immunotherapies in cancer treatment increases the risk of immunotherapy-related cardiotoxicity. In contrast to conventional chemotherapy, these novel therapies have expanded the forms and presentations of cardiovascular damage to a broad spectrum from asymptomatic changes to fulminant short- and long-term complications in terms of cardiomyopathy, arrythmia, and vascular disease. In cancer patients and, particularly, cancer patients undergoing (immune-)therapy, cardio-oncological monitoring is a complex interplay between pretherapeutic risk assessment, identification of impending cardiotoxicity, and post-therapeutic surveillance. For these purposes, the cardio-oncologist can revert to a broad spectrum of nuclear cardiological diagnostic workup. The most promising commonly used nuclear medicine imaging techniques in relation to immunotherapy will be discussed in this review article with a special focus on the continuous development of highly specific molecular markers and steadily improving methods of image generation. The review closes with an outlook on possible new developments of molecular imaging and advanced image evaluation techniques in this exciting and increasingly growing field of immunotherapy-related cardiotoxicity.


Astrocytes regulate the balance between plasminogen activation and plasmin clearance via cell-surface actin.
Plasminogen activation is involved in many processes within the central nervous system, including synaptic plasticity, neuroinflammation and neurodegeneration. However, the mechanisms that regulate plasminogen activation in the brain still remain unknown. Here we demonstrate that astrocytes participate in this regulation by two mechanisms. First, the astrocyte plasma membrane serves as a surface for plasminogen activation by tissue-type plasminogen activator. This activation triggers downstream plasmin-dependent processes with important impacts in brain health and disease, such as fibrinolysis and brain-derived neurotrophic factor conversion. Second, astrocytes take up plasminogen and plasmin in a regulated manner through a novel mechanism involving endocytosis mediated by cell-surface actin and triggered by extracellular plasmin activity at the surface of astrocytes. Following endocytosis, plasminogen and plasmin are targeted to lysosomes for degradation. Thus, cell-surface actin acts as a sensor of plasmin activity to induce a negative feedback through plasmin endocytosis. This study provides evidence that astrocytes control the balance between plasmin formation and plasmin elimination in the brain parenchyma.


Epigenetic Programming by Early-Life Stress.
Studies show that adverse conditions during early life can increase risks of developing mood disorders later in life. It is currently hypothesized that levels of environmental adversity in this early developmental period are able to shape the experience-dependent maturation of stress-regulating pathways leading to long-lasting alterations in stress responsivity during adulthood; a phenomenon often referred to as "early-life programming." Research is addressing the molecular mechanisms underlying this programming by which gene-environment interactions can predispose individuals toward psychopathology. Here we review key findings from animal and clinical studies examining the effects of prenatal and postnatal environment in shaping development of the neuroendocrine regulation of stress and the role of epigenetic mechanisms in translating early-life conditions into long-lasting gene expression changes underpinning stress-related behaviors. We also review latest ideas regarding interactions between environments at different developmental stages and evidence for transgenerational effects. Understanding how prenatal and postnatal experiences can give rise to lasting epigenetic marks conferring increased vulnerability to mental disorders is a major focus of molecular psychiatry and should pave new guidelines for therapeutic interventions.


[Positron emission tomography in diagnosis and prognosis of postanoxic cerebral dysfunctions].
Clinical signs, laboratory tests, EEG or evoked potentials only permit an indirect estimation of the extent of structural brain damage following severe global brain anoxia. Positron emission tomography (PET) permits additional insights into the extent of neuronal damage in acute and persistent postanoxic vegetative states (VS). PET documents a severe and irreversible damage of supratentorial cortical structures in postanoxic VS and allows its differential diagnosis from related disorders, e.g. the locked-in-syndrome. PET clearly distinguishes functional alterations in VS from those in non-REM sleep and documents that patients in VS are not in a sleep-like condition, but in a state closely related to deep anesthesia. Furthermore, the extent of impairment of the residual cortical glucose consumption yields information concerning the possible recovery of consciousness and neuronal function in VS. In combination with clinical, laboratory and neurophysiological findings, PET may be helpful to establish the individual prognosis in acute VS.


Enterotoxaemia in goats.
Enterotoxaemia of sheep and goats occurs worldwide, but the condition in goats is poorly understood. The disease in goats is mostly caused by Clostridium perfringens type D, although the role of the toxins of this microorganism in the pathogenesis of the disease is not fully understood. The disease occurs in three forms, peracute, acute and chronic, the cardinal clinical sign of the acute and chronic forms being diarrhoea. The main biochemical alterations are hyperglycaemia and glycosuria, while at necropsy the disease is often characterized by haemorrhagic colitis. The typical histological changes observed in the brain of sheep with enterotoxaemia are not considered to be a common feature of enterotoxaemia in goats. Although the pathogenesis of caprine enterotoxaemia has not yet been properly defined, it is usually accepted that the presence of C. perfringens type D in the small bowel, together with a sudden change to a diet rich in carbohydrates, is the main predisposing factor for the disease. Vaccination seems to be poorly effective in preventing caprine enterotoxaemia, which might be due to the fact that the enteric form of the disease is partially independent of circulating C. perfringens toxin. More studies are needed on caprine enterotoxaemia, especially of its pathogenesis and immunity, in order to develop more efficient control measures for this disease.


Microsatellite DNA analyses of a highly disjunct New Zealand tree reveal strong differentiation and imply a formerly more continuous distribution.
Although New Zealand is a biodiversity hotspot, there has been little genetic investigation of why so many of its threatened and uncommon plants have naturally disjunct distributions. We investigated the small tree Pseudopanax ferox (Araliaceae), which has a widespread but highly disjunct lowland distribution within New Zealand. Genotyping of nuclear microsatellites and a chloroplast locus revealed pronounced genetic differentiation and four principal genetic clusters. Our results indicate that the disjunct distribution is a product of vicariance rather than long-distance dispersal. This highlights the need to preserve multiple populations when disjunct distributions are the result of vicariance, rather than focusing conservation efforts on a core area, in order to retain as much as possible of a species' evolutionary legacy and potential. Additionally, based on our genetic findings and the ecology of P. ferox, we hypothesize that it was more continuously distributed during the drier (but not maximally colder) interstadials of glacial periods and/or on the fertile soils available immediately postglacial. We further hypothesize that P. ferox belongs to a suite of species of drought-prone and/or fertile habitats whose distributions are actually restricted during warmer and wetter interglacial periods, despite being principally of the lowlands. Our genetic data for P. ferox are also the first consistent with the survival during the Last Glacial Maxima of a lowland tree at high latitudes in the south-eastern South Island.


Effects of exercise on longevity of rats.
It has been postulated that life span is inversely related to energy expenditure. If this is correct, regularly performed exercise could accelerate the aging process. In two early studies, exercise shortened the life span of rats; the results of these studies have been cited as evidence for the concept that an increase in energy expenditure accelerates aging. However, subsequent studies have not confirmed this finding. Instead, the weight of evidence now indicates that rats that exercise regularly have a longer average life span than sedentary, ad libitum-fed controls. Freely eating sedentary rats become obese, indicating that their food intake is in excess of their energy requirements. Available evidence seems compatible with the interpretation that exercise results in improved survival in rats by countering deleterious effects of a sedentary life combined with overeating.


Limited proteolysis and proton NMR spectroscopy of Bacillus stearothermophilus pyruvate dehydrogenase multienzyme complex.
The pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus was treated with chymotrypsin at pH 7 and 0 degrees C. Loss of the overall catalytic activity lagged behind the rapid cleavage of the lipoate acetyltransferase polypeptide chains, whose apparent Mr fell from 57 000 to 45 000 as judged by sodium dodecylsulphate/polyacrylamide gel electrophoresis. The inactive chymotrypsin-treated enzyme had lost the lipoic-acid-containing regions of the lipoate acetyltransferase chains, yet remained a highly assembled structure. Treatment of this chymotryptic core complex with trypsin at pH 7.0 and 0 degrees C caused a further shortening of the lipoate acetyltransferase polypeptide chains to an apparent Mr of 28 000 and was accompanied by disassembly of the complex. The lipoic-acid-containing regions are therefore likely to be physically exposed in the intact complex, protruding from the structural core formed by the lipoate acetyltransferase component between the subunits of the other component enzymes. Proton nuclear magnetic resonance spectroscopy demonstrated that the enzyme complex contains large regions of polypeptide chain with remarkable intramolecular mobility, most of which were retained after excision of the lipoic-acid-containing regions with chymotrypsin. It is likely that the highly mobile regions are in the lipoate acetyltransferase component and facilitate movement of the lipoic acid residues. Such polypeptide chain mobility provides the molecular basis of a novel system of active-site coupling in the 2-oxo acid dehydrogenase multienzyme complexes.


Genomic insights into host and parasite interactions during intracellular infection by Toxoplasma gondii.
To gain insights into the molecular interactions of an intracellular pathogen and its host cell, we studied the gene expression and chromatin states of human fibroblasts infected with the Apicomplexan parasite Toxoplasma gondii. We show a striking activation of host cell genes that regulate a number of cellular processes, some of which are protective of the host cell, others likely to be advantageous to the pathogen. The simultaneous capture of host and parasite genomic information allowed us to gain insights into the regulation of the T. gondii genome. We show how chromatin accessibility and transcriptional profiling together permit novel annotation of the parasite's genome, including more accurate mapping of known genes and the identification of new genes and cis-regulatory elements. Motif analysis reveals not only the known T. gondii AP2 transcription factor-binding site but also a previously-undiscovered candidate TATA box-containing motif at one-quarter of promoters. By inferring the transcription factor and upstream cell signaling responses involved in the host cell, we can use genomic information to gain insights into T. gondii's perturbation of host cell physiology. Our resulting model builds on previously-described human host cell signalling responses to T. gondii infection, linked to induction of specific transcription factors, some of which appear to be solely protective of the host cell, others of which appear to be co-opted by the pathogen to enhance its own survival.


Incidence of concussion and recovery of neurocognitive dysfunction on ImPACT assessment among youth athletes with premorbid depression or anxiety taking antidepressants.
Concussions in youth sports comprise an estimated 1.6-3.8 million annual injuries in the US. Sex, age, and attention-deficit hyperactivity disorder (ADHD) have been identified as salient risk factors for concussion. This study seeks to evaluate the role of premorbid depression or anxiety (DA), with or without antidepressant use, on the incidence of concussion and the recovery of symptoms and neurocognitive dysfunction after concussion. Immediate Postconcussion Assessment and Cognitive Testing (ImPACT) was administered to 7453 youth athletes at baseline. Throughout the season, concussions were examined by physicians and athletic trainers, followed by readministration of ImPACT postinjury (PI) and again at follow-up, a median of 7 days PI. Individuals were divided into three categories: 1) unmedicated athletes with DA (DA-only, n = 315), athletes taking antidepressants (DA-meds, n = 81), and those without DA or antidepressant use (non-DA, n = 7039). Concussion incidence was calculated as the total number of concussions per total number of patient-years. The recovery of neurocognitive measures PI was calculated as standardized deviations from baseline to PI and then follow-up in the 5 composite ImPACT scores: symptom score, verbal memory, visual memory, visual motor skills, and reaction time. Univariate results were confirmed with multivariate analysis. There was no difference in concussion incidence between the DA-only cohort and the non-DA group. However, the DA-meds group had a significantly greater incidence of concussion than both the DA-only group (OR 2.67, 95% CI 1.88-7.18, p = 0.0001) and the non-DA group (OR 2.19, 95% CI 1.16-4.12, p = 0.02). Deviation from baseline in PI symptom scores was greater among the DA-meds group as compared to the non-DA group (OR 1.14, 95% CI 1.01-1.28, p = 0.03). At follow-up, the deviation from baseline in symptom scores remained elevated among the DA-meds group as compared to the non-DA group (OR 1.62, 95% CI 1.20-2.20, p = 0.002) and the DA-only group (OR 1.87, 95% CI 1.12-3.10, p = 0.02). Deviation from baseline in follow-up verbal memory was also greater among the DA-meds group as compared to both the non-DA group (OR 1.57, 95% CI 1.08-2.27, p = 0.02) and the DA-only group (OR 1.66, 95% CI 1.03-2.69, p = 0.04). Premorbid DA itself does not seem to affect the incidence of concussion or the recovery of symptoms and neurocognitive dysfunction PI. However, antidepressant use for DA is associated with 1) increased concussion incidence and 2) elevated symptom scores and verbal memory scores up to 7 days after concussion, suggesting impaired symptomatic and neurocognitive recovery on ImPACT.


The LysR-type transcriptional regulator CysB controls the repression of hslJ transcription in Escherichia coli.
The LysR-type transcriptional regulator (LTTR) CysB is a transcription factor in Escherichia coli cells, where as a homotetramer it binds the target promoter regions and activates the genes involved in sulphur utilization and sulphonate-sulphur metabolism, while negatively autoregulating its own transcription. The hslJ gene was found to be negatively regulated by CysB and directly correlated with novobiocin resistance of the bacterium. cysB mutants showed upregulation of the hslJ : : lacZ gene fusion and exhibited increased novobiocin resistance. In this study the hslJ transcription start point and the corresponding putative sigma(70) promoter were determined. The hslJ promoter region was defined by employing different hslJ-lacZ operon fusions, and transcription of the hslJ gene was shown to be subject to both repression imposed by the CysB regulator and direct or indirect autogenous negative control. These two regulations compete to some extent but they are not mutually exclusive. CysB acts as a direct repressor of hslJ transcription and binds the hslJ promoter region that carries the putative CysB repressor site. This CysB binding, apparently responsible for repression, is enhanced in the presence of the ligand N-acetylserine (NAS), hitherto considered to be a positive cofactor in CysB-mediated gene regulations. Interallelic complementation of characterized CysB mutants I33N and S277Ter partially restored the repression of hslJ transcription and the consequent novobiocin sensitivity, but did not complement the cysteine auxotrophy.