Effects of maternal smoking and exposure to methylmercury on brain-derived neurotrophic factor concentrations in umbilical cord serum.
Brain-derived neurotrophic factor (BDNF) is a neurotrophin essential for neuronal survival and differentiation. We examined the concentration of BDNF in cord serum from newborns exposed to methylmercury (MeHg) and polychlorinated biphenyls (PCB) in utero by maternal consumption of whale meat. The cohort consisted of 395 singleton births (206 boys and 189 girls), gestational age ranging from 38 to 42 weeks. Serum BDNF was measured by sandwich ELISA. Maternal smoking habits and other relevant factors were obtained by interviewing the mothers. The exposure to MeHg was estimated from Hg concentrations in cord blood, whereas exposure to PCB was estimated based on maternal serum concentrations. Only MeHg exposure affected the serum BDNF, which decreased in a concentration-dependent manner in girls born to nonsmoking mothers. Maternal smoking significantly increased BNDF in girls but not in boys. For further statistical analyses, we used the serum BDNF concentration as a continuous outcome variable in supervised regression models. Serum BDNF concentration increased with gestational age, increased by maternal smoking, decreased slightly with MeHg exposure, and maternal smoking enhanced the decrease in serum BDNF induced by MeHg exposure. Cord blood BDNF has been reported to increase in association with perinatal brain injuries and has been proposed as a possible predictive marker of neurodevelopmental outcomes. The negative effect that MeHg seems to exert on cord blood BDNF concentration could endanger compensatory responses to an adverse impact and therefore deserves attention.


Functional neuroimaging of the Iowa Gambling Task in older adults.
The neural systems most susceptible to age-related decline mirror the systems linked to decision making. Yet, the neural processes underlying decision-making disparities among older adults are not well understood. We sought to identify neural response patterns that distinguish 2 groups of older adults who exhibit divergent decision-making patterns. Participants were 31 healthy older adults (ages 59-88, 53% female), defined as advantageous or disadvantageous decision-makers based on Iowa Gambling Task (IGT) performance, who completed an alternate version of the IGT while undergoing functional MRI. The groups were indistinguishable on neuropsychological testing. We contrasted the BOLD signal between groups during 3 phases of the decision-making process: Prechoice (preselection), Prefeedback (postselection), and Feedback (receipt of gains/losses). We further examined whether BOLD signal varied as a function of age in each group. We observed greater activation among the IGT-Disadvantageous relative to -Advantageous older adults in the prefrontal cortex during the early phases of the decision-making process (Prechoice), and in posterior brain regions (e.g., the precuneus) during the later phases (Prefeedback and Feedback). We also found that with increasing age, IGT-Advantageous older adults showed increasing activation in the prefrontal cortex during all phases and increasing activation in the posterior cingulate during earlier phases of the decision process. By contrast, the IGT-Disadvantageous older adults exhibited a reduced or reversed trend. These functional differences may be a consequence of altered reward processing or differing compensatory strategies between IGT-Disadvantageous and -Advantageous older adults. This supports the notion that divergent neurobiological aging trajectories underlie disparate decision-making patterns.


Phase I/II study of 3TC (lamivudine) in HIV-positive, asymptomatic or mild AIDS-related complex patients: sustained reduction in viral markers. The Lamivudine European HIV Working Group.
To evaluate the efficacy of 3TC (lamivudine), a synthetic nucleoside analogue that inhibits HIV reverse transcriptase in vitro, as treatment for HIV-positive, asymptomatic or mild AIDS-related complex patients. Open-label, multinational and multicentre, non-comparative, escalating dose study. Patients who meet the selection criteria (n = 104) were enrolled in three European countries. Ten to 15 patients were included at each of the six dose levels of 3TC (0.5, 1.0, 2.0, 4.0, 8.0, 12.0 and 20.0 mg/kg daily in two divided doses every 12 h). Virological parameters--immune-complex dissociation (ICD) assay for HIV p24 antigenaemia, plasma HIV RNA load, whole blood assay and cellular viraemia--were evaluated at weeks 0, 4, 12 and 24. Sustained reductions in HIV RNA load and in ICD p24 antigen levels were observed and maintained over the 12-week assessment period. Greater reductions were noted at higher doses but this trend did not reach statistical significance. In 38 patients, reductions of cell viraemia were significantly greater at 4 weeks for patients treated at higher doses of 3TC. These virological data show that 3TC is a potent inhibitor of HIV replication in HIV-positive, asymptomatic or mild ARC patients as assessed by ICD p24 antigenaemia, plasma HIV RNA load and cell viraemia.


Left ventricular diastolic performance at rest in patients with angina and normal systolic function--assessment by equilibrium radionuclide angiography.
The aim of the study was to correlate diastolic function, as evaluated by peak filling rate (PFR) and relative time (TPFR), with the severity of ischemic heart disease, as evaluated by exercise electrocardiography. Accordingly, 83 ischemic patients with effort angina, but normal ejection function at rest and normal left ventricular size, were studied by equilibrium radionuclide angiocardiography within two weeks from the exercise ECG. Diastolic dysfunction, as determined from PFR and, to a lesser extent, from TPFR, is common in patients with ischemic heart disease and normal systolic function. The prevalence and severity of such dysfunction is related more to the severity of the ischemia, as evaluated by the exercise ECG, than to the presence of an old myocardial infarction. Such findings are consistent with the hypothesis that PFR reflects mainly the early diastolic active uncoupling process.


A resilient battery electric bus transit system configuration.
Electric mobility is fundamental to combat climate change and attaining the United Nations Sustainable Development Goals (SDG-11). However, electric mobility necessitates a seamless integration between power and transportation systems, as the resiliency of both systems is becoming far more interdependent. Here, we focus on disruption to Battery Electric Bus (BEB) transit system charging infrastructure and offer a resilient BEB transit system planning model. The proposed model optimizes the BEB system costs while ensuring the system's robustness against simultaneous charging station failures. In our case study, a single charging station failure would lead to up to 34.03% service reduction, and two simultaneous failures would reduce the service by up to 58.18%. Our proposed two-stage robust model addresses this issue with a relatively small added cost (3.26% and 8.12% higher than the base model). This cost enables uninterrupted BEB system operation during disruption, ensuring personal mobility, social interaction, and economic productivity.


Permissible idealizations for the purpose of prediction.
Every model leaves out or distorts some factors that are causally connected to its target phenomenon-the phenomenon that it seeks to predict or explain. If we want to make predictions, and we want to base decisions on those predictions, what is it safe to omit or to simplify, and what ought a causal model to describe fully and correctly? A schematic answer: the factors that matter are those that make a difference to the target phenomenon. There are several ways to understand differencemaking. This paper advances a view as to which is the most relevant to the forecaster and the decision-maker. It turns out that the right notion of differencemaking for thinking about idealization in prediction is also the right notion for thinking about idealization in explanation; this suggests a carefully circumscribed version of Hempel's famous thesis that there is a symmetry between explanation and prediction.


Novel compounds in the Zr-O system, their crystal structures and mechanical properties.
With the motivation of exploring new high-strength ceramics, ab initio evolutionary simulations are performed to search for all the stable compounds in the Zr-O system. We have found that not only the traditional compound ZrO2, but also the ordered suboxides R3̅-Zr6O, R3̅c-Zr3O, P3̅1m-Zr2O and P6̅2m-ZrO are stable at zero pressure. The crystal structure of semimetallic P6̅2m-ZrO consists of Zr-graphene layers and can be described as an intercalated version of the ω-Zr structure. An interesting massive Dirac cone is found in the three-dimensional (3D) band structure of P6̅2m-ZrO at the Γ-point. The elastic properties, the hardness and the correlation between the mechanical properties of Zr-O compounds and the oxygen content have been systematically investigated. Surprisingly, the hardest zirconium oxide is not ZrO2, but ZrO. Both P6̅2m-ZrO and P3̅1m-Zr2O exhibit relatively high hardness values of 14 GPa and 10 GPa, respectively. The anisotropic Young's modulus E, torsion shear modulus Gt and linear compressibility β have been derived for P6̅2m-ZrO and P3̅1m-Zr2O. Further analyses of the density of states, the band structure and the crystal orbital Hamilton population indicate that the electronic structure of Zr-O compounds is directly related to their mechanical properties. The simultaneous occurrence of the 3D-framework of Zr-O and the strong Zr-Zr bonds in P6̅2m-ZrO explains its high hardness.


Probiotic Lactobacillus rhamnosus GG Promotes Mouse Gut Microbiota Diversity and T Cell Differentiation.
Lactic acid bacteria (LAB) are the primary genera of the intestinal flora and have many probiotic functions. In the present study, Lactobacillus rhamnosus GG (LGG) ATCC 53103 was used to treat BALB/c mice. After LGG intervention, both low and high LGG doses were shown to improve the observed OTU, Chao1, ACE, and Shannon indices, while the Simpson index decreased, demonstrating that LGG can promote intestinal microbiota abundance and diversity. Furthermore, LGG treatment increased the abundances of intestinal Firmicutes, Bacteroides and Actinomycetes while reducing that of Proteobacteria. In addition to its effect on gut the microbiota, LGG could also regulate the host immune system. In the present study, we showed that LGG could affect the percentage of CD3+ T lymphocytes in the spleens (SPLs), mesenteric lymph nodes (MLNs), Peyer's patches (PPs) and lamina propria lymphocytes (LPLs) of mice, including total CD3+ T, CD3+CD4+ T, and CD3+CD8+ T lymphocytes. Furthermore, LGG could effectively increase the expression of Th1-type cytokines (IFN-γ) and Th2 cytokines (IL-4) in CD4+ T cells, indicating that the proportion of Th1 and Th2 cells in mice with LGG treatment was in a high equilibrium state compared to the control group. In addition, the IFN-γ/IL-4 ratio was greater than 1 in mice with LGG intervention, suggesting that LGG tends to mediate the Th1 immune response. The results of the present study also showed that LGG upregulated the expression of IL-17 in CD4+ T cells and regulated the percentage of CD4+CD25+Foxp3+ Treg cells in various secondary immunological organs, indicating that LGG may promote the balance of Th-17 and Treg cells.


Hyperglycemia alters PI3k and Akt signaling and leads to endothelial cell proliferative dysfunction.
Diabetes mellitus is a major risk factor for the development of vascular complications. We hypothesized that hyperglycemia decreases endothelial cell (EC) proliferation and survival via phosphatidylinositol 3-kinase (PI3k) and Akt signaling pathways. We cultured human umbilical vein ECs (HUVEC) in 5, 20, or 40 mM d-glucose. Cells grown in 5, 20, and 40 mM mannitol served as a control for osmotic effects. We measured EC proliferation for up to 15 days. We assessed apoptosis by annexin V and propidium iodide staining and flow cytometry, analyzed cell lysates obtained on culture day 8 for total and phosphorylated PI3k and Akt by Western blot analysis, and measured Akt kinase activity using a GSK fusion protein. HUVEC proliferation was also tested in the presence of pharmacological inhibitors of PI3k-Akt (wortmannin and LY294002) and after transfection with a constitutively active Akt mutant. ECs in media containing 5 mM d-glucose (control) exhibited log-phase growth on days 7-10. d-Glucose at 20 and 40 mM significantly decreased proliferation versus control (P < 0.05 for both), whereas mannitol did not impair EC proliferation. Apoptosis increased significantly in HUVEC exposed to 40 mM d-glucose. d-Glucose at 40 mM significantly decreased tyrosine-phosphorylated PI3k, threonine 308-phosphorylated-Akt, and Akt activity relative to control 5 mM d-glucose. Pharmacological inhibition of PI3k-Akt resulted in a dose-dependent decrease in EC proliferation. Transfection with a constitutively active Akt mutant protected ECs by enhancing proliferation when grown in 20 and 40 mM d-glucose. We conclude that d-glucose regulates Akt signaling through threonine phosphorylation of Akt and that hyperglycemia-impaired PI3k-Akt signaling may promote EC proliferative dysfunction in diabetes.


Tissue-specific distribution of cross-linked somatostatin receptor proteins in the rat.
Pharmacological studies have suggested that the somatostatin (SS) receptor is heterogeneous and exhibits SS-14-and SS-28-selective subtypes. Whether such subtypes arise from molecular heterogeneity of the receptor protein has not been definitively established. Previous reports characterizing the molecular properties of the SS receptor by the cross-linking approach have yielded divergent size estimates ranging from 27 kDa to 200 kDa. In order to resolve this discrepancy, as well as to determine whether SS-14 and SS-28 interact with specific receptor proteins, we have cross-linked radioiodinated derivatives of [125I-Tyr11]SS-14 (T*-SS-14) and [Leu8,D-Trp22,125I-Tyr25]SS-28 (LTT*-SS-28) to membrane SS receptors in rat brain, pituitary, exocrine pancreas and adrenal cortex using a number of chemical and photoaffinity cross-linking agents. The labelled cross-linked receptor proteins were analysed by SDS/PAGE under reducing conditions followed by autoradiography. Our findings indicate that the pattern of specifically labelled cross-linked SS receptor proteins is sensitive to the concentration of chemical cross-linking agents such as disuccinimidyl suberate and dithiobis-(succinimidyl propionate). Labelled high-molecular-mass complexes of cross-linked receptor-ligand proteins were observed only when high concentrations of these cross-linkers were employed. Using optimized low concentrations of cross-linkers, however, two major labelled bands of 58 +/- 3 kDa and 27 +/- 2 kDa were detected. These two bands were identified as specifically labelled SS receptor proteins subsequent to cross-linking with a number of photoaffinity cross-linking agents as well. We demonstrate here that the 58 kDa protein is the major SS receptor protein in the rat pituitary, adrenal and exocrine pancreas, whereas the 27 kDa moiety represents the principal form in the brain. Additionally, the presence of a minor specifically labelled band of 32 kDa was detected uniquely in the brain, and a minor labelled protein of 42 kDa was observed in the pancreas. The labelling pattern obtained with LTT*-SS-28 was identical to that observed with T*-SS-14. Labelling of the 27 kDa band by either ligand was inhibited by SS-14 and SS-28 in a dose-dependent manner. Densitometric quantification showed that SS-14 exhibited greater than 2-fold greater potency than SS-28 for inhibiting the labelling of the 27 kDa species. These findings emphasize the need for careful interpretation of cross-linking data obtained for SS receptors, and provide evidence for molecular heterogeneity and for a tissue-specific distribution of the two principal SS receptor proteins.


[Detection of BCR-ABL Fusion Gene in Chronic Myeloid Leukemia by Novel Digital PCR].
To establish a new digital polymerase chain reaction (dPCR) system for the detection of BCR-ABL fusion gene in patients with chronic myeloid leukemia (CML), and explore its analytical performance and clinical applicability in the detection of BCR-ABLp190/210/230. A new dPCR system for detecting BCR-ABLp190/210/230 was successfully developed, and its sensitivity difference with qPCR and improvement of drug side effects in patients with CML during drug reduction or withdrawal were compared. Among 176 samples, qPCR and dPCR showed high consistency in the sensitivity of detecting BCR-ABL (82.39%), and the positive rate of dPCR was about 5 times higher that of qPCR (20.45% vs 3.98%). During follow-up, blood routine (25% vs 10%), kidney/liver/stomach (25% vs 20%) and cardiac function (10% vs 0) were significantly improved after drug reduction or withdrawal in patients with initial dPCR negative compared with before drug reduction or withdrawal. This new dPCR detection system can be applied to the detection of BCR-ABLp190/210/230. It has better consistency and higher positive detection rate than qPCR. Drug withdrawal or dose reduction guided by dPCR has a certain effect on improving drug side effects.


Comparison of Calorie and Protein Intake of Very Low Birth Weight Infants Receiving Mother's Own Milk or Donor Milk When the Nutrient Composition of Human Milk Is Measured With a Breast Milk Analyzer.
In premature infants, donor breast milk (DBM) is assumed to provide reduced nutrients vs. mother's own milk (MOM). This study examined calorie and protein delivery when very low birth weight infants were fed fortified MOM or DBM, with a known nutrient composition, relative to established nutrition recommendations and to determine if there were differences between the groups. A retrospective medical record review was conducted in 29 very low birth weight infants receiving MOM or DBM. Nutrient content of human milk was measured using the Calais Analyzer. Added fortifiers feeding volume, and weight were collected to determine total daily calorie and protein intake. 145 days of enteral feedings among 29 infants were included, 78 (53.8%) from DBM and 67 (46.2%) from MOM. Mean daily fluid intake among infants receiving DBM was significantly higher when compared with MOM, 150.6 ± 7.6 mL/kg vs 146.8 ± 11.3 mL/kg (P = .016). DBM feedings provided 110.1 ± 9.0 kcals/kg/d vs 113.0 ± 21.0 kcals/kg/d from MOM feedings (P = .275). Mean protein intake was similar, 4.1 ± 0.5 g /kg/d on DBM days vs 4.0 ± 0.5 g kg/d on MOM days (P = .162). A total of 46 of 78 DBM days (59.0%) and 30 of 67 MOM days (44.8%) were below the minimum established calorie needs of 110 kcals/kg/day. DBM provides comparable nutrient intake to MOM at a higher enteral feeding volume. However, both types of human milk failed to meet energy needs with standard fortification regimens.


Web versus press with a high circulation: an Italian case study.
the quality of health-related information available for general public on the web has been widely debated, often expressing concern, in the last few years, but also other mass media supply medical contents for health consumer. To evaluate the quality of information available by the Italian people through Internet in comparison with the one published on the press with a high circulation. A general interest topic such as over-weight and obesity has been identified as a convenient field for the comparison. A sample of web sites and articles have been collected--excluding the promotional ones- in order to compare their completeness and correctness. National guidelines were the golden standard. For the web sites, the availability of indications to let the user know the reliability of the site--we will refer to these as "transparency"--has been evaluated through 9 parameters. 45 informative sites and 35 articles have been found: the Italian population has a good chance of getting information. Web sites are much more complete than articles. Correctness of both is quite good. Transparency of web sites leaves much to desire, independently from the quality of the content. There is probably an excessive confidence about the transparency and caution of articles in dealing with health-related topics. The web publishers must necessarily be educated to apply the existing good practice rules, but in the meantime also the press with a high circulation, that is read by a high percentage of people, has to improve its quality in order to co-operate in supplying adequate information. In general, from the traditional exceptionalist approach for medical information on the internet, a definition of media quality standard could be derived.


Structure and function of Cas-L, a 105-kD Crk-associated substrate-related protein that is involved in beta 1 integrin-mediated signaling in lymphocytes.
Integrin/ligand binding evokes tyrosine phosphorylation of various proteins. We reported previously that a 105 kD protein (pp105) was tyrosine phosphorylated by the engagement of beta 1 integrins in T lymphocytes. We show here that pp105 is a novel p130Cas (Crk-associated substrate)-related protein. Deduced amino acid sequence revealed that pp105 contains conserved motifs with p130Cas, and both pp105 and p130Cas bind to focal adhesion kinase (pp125FAK) and Crk. However, pp105 has a clearly distinct structure from p130Cas, and pp105 is preferentially expressed in lymphocytes, whereas p130Cas is expressed in adherent cells. With these findings, we designate pp105 as Cas-L, lymphocyte-type Cas. Furthermore, we demonstrate that integrin/ligand binding results in the recruitment of Crk, Nck, and SHPTP2 to pp105. These findings further define the roles of pp105/Cas-L and pp125FAK in the integrin-mediated signaling pathways.


Fetal hemoglobin and F-cell responses to long-term hydroxyurea treatment in young sickle cell patients. The French Study Group on Sickle Cell Disease.
We have studied the cellular and molecular responses to long-term hydroxyurea (HU) treatment in 29 severely affected young patients with sickle cell disease (mean age, 10.9 +/- 4.1 years). Patients received HU at 20 mg/kg/d on 4 consecutive days per week initially, with a monthly escalated dose avoiding marrow-toxicity (mean steady-state dose, 34.2 +/- 4.6 mg/kg/d) for 12 to 36 months (mean duration, 22 months). The studied parameters were hemoglobin F (HbF), F reticulocytes (F retics), F cells, the amount of HbF per F cell (F/F cell), polymer tendency at 40% and 70% oxygen saturation, and hemolysis. Initial HbF (Fi) was dispersed (from 0.85% to 13.9%). HbF increased in all patients but 1. HbF at maximal response (Fmax) reached a sustained level varying from a 1.5-fold to a 16-fold Fi after a variable delay (6 to 24 months). Fmax was not related to HU dosage, but triangle upF (Fmax - Fi) was strongly correlated to triangle upMCV (MCVmax - MCVi). HbF increase resulted from the increase of both F cells and F/F cell. In this rather short series, Fi and Fmax were not significantly associated with age, gender, or beta-globin haplotype. Neither Fmax nor triangle upF was related to bone marrow reserve, as measured by baseline reticulocyte or neutrophil counts. However, Fmax was highly dependent on Fi. When patients are individualized into three groups according to Fmax (group 1, Fmax >20% [12 patients]; group 2, 10% < Fmax < 20% [11 patients]; group 3, Fmax <10% [5 patients]), Fi is significantly different between groups, being the highest in group 1. In addition, the best responders (group 1) were significantly different from patients in the two other groups with higher levels of total hemoglobin, decreased bilirubin, and decreased polymer tendency.


[Cardiovascular complications of pancreatis diseases].
Under physiological conditions, the pancreas scarcely influences the function of the cardiovascular system, although the hormones produced in the healthy pancreas (insulin, glucagon and somatostatin) affect the myocardial contractility in pharmacological doses. Among the diseases of the pancreas, the pancreatic tumours (insulinoma, glucagonoma and vipoma), furthermore the acute and chronic pancreatitis involve cardiovascular complications, which influence the outcome of the disease. Although the clinical picture is dominated by the metabolic changes of the excessively produced hormones in pancreatic tumours, the cardiac and vascular effects of the hormones may be considerable. In acute necrotizing pancreatitis, enzymes released from the pancreas and inflammatory mediators transform acute necrotizing pancreatitis into "multiple organ disease"; one of the important forms of this disease is the cardiovascular shock syndrome. One of the best-known complications of chronic pancreatitis is the pancreoprive diabetes mellitus, and beside that other, nonspecific cardiac alterations (e.g. ECG-changes) may occur.


Impact of plasma histones in human sepsis and their contribution to cellular injury and inflammation.
Circulating histones have been identified as mediators of damage in animal models of sepsis and in patients with trauma-associated lung injury. Despite existing controversies on actual histone concentrations, clinical implications and mechanism of action in various disease conditions, histone levels in human sepsis, association with disease progression and mediated effects on endothelial and immune cells remain unreported. This study aimed to determine histone levels and its clinical implication in septic patients and to elucidate histone-mediated effects ex-vivo. Histone levels, endogenous activated protein C (APC) levels and clinical data from two independent cohorts of septic patients were obtained. Histone levels were compared with various control groups including healthy individuals, intensive care unit (ICU) patients without sepsis, ICU patients with multiple organ failure and patients with minor or multiple trauma, all without infection. Endothelial and monocytic cells were stimulated with histones. Cellular integrity and sepsis prototypical cytokines were evaluated. The mechanism of action of histones via Toll-like receptor 4 (TLR4) was evaluated using a function blocking antibody. Histone degradation in plasma was studied by immunoblotting. Histone H4 levels were significantly elevated in patients with sepsis (cohort I; n = 15 and cohort II; n = 19) versus ICU controls (n = 12), patients with multiple organ failure (n = 12) or minor trauma (n = 7), associated with need for renal replacement therapy and decrease in platelet count during disease progression, and remarkably were significantly associated with increased mortality rates in septic patients (ICU-, 28 day- and 90 day mortality rates). There was an inverse correlation between plasma histones and endogenous APC levels. Histone stimulation induced the release of sepsis prototypic cytokines and decreased cell integrity indicated by a significant increase of lactate dehydrogenase (LDH) and propidium iodide (PI) staining. Blocking of TLR4 decreased cellular cytotoxicity on endothelial cells. The calculated half-life of histones in spiked plasma was 4.6 minutes. Histone levels in septic patients are significantly increased and might mediate disease aggravation by cellular injury and inflammation via TLR4 signaling, which potentially results in multiple organ failure and fatal outcome.


[Pyruvate- and alpha-ketoglutarate dehydrogenase activity during yeast growth on glucose and hexadecane].
The activity of pyruvate- and ketoglutarate dehydrogenases (PDH and KGDH) was determined in the cells of Candida lipolytica and Candida tropicalis during the exponential phase of their growth on glucose and hexadecane. The activity of PDH in the cells growing on hexadecane was found to be lower than in the cells growing on glucose. The activity of KGDH was almost the same in the cells cultivated on hexadecane and glucose. The results obtained are discussed with respect to different pathways of degradation of glucose and n-alkanes, and a decreased requirement in thiamine during growth of the yeast cells on n-alkanes.


[Neurotensin influence on painful stress afteractions in rats with neurotoxic damages of serotoninergic structures of brain substantia nigra].
The purpose of the research was to reveal the features of neurotensin influence on behavior of rats with damages of 5-HT structures of substantia nigra. Changes of recall of passive avoidance conditioned reactions, and also painful stimulation aftereffects on locomotor activity of rats in "open field" were studied. It was shown that neurotoxin 5,7-DOT administration into substantia nigra impaired recall of passive avoidance reactions and as well weakened oppressive aftereffects of painful stimulation. Administration of serotonin 5-HT1A receptors antagonist p-MPPF insert similar influence on aftereffects of painful stimulation. Neurotensin microinjections into caudate nucleus just before painful stimulation prevented disturbances of defensive behavior and its aftereffects evoked by neurotoxin. Neurotensin administration into substantia nigra in 24 h after painful stimulation didn't exert any significant influence on passive avoidance reactions but increased motor activity against a background of its recall. Effects of neurotoxin administrations into substantia nigra connected with weakened of painful stress influence on motor activity in rats. The prevention of this effect development after neurotensin administrations into caudate nucleus may be specified by recovery of recall passive avoidance reactions destroyed by neurotoxin action and is explained by normalization of relationships balance of 5-HT and dopaminergic brain systems.


OrthoMaM v12: a database of curated single-copy ortholog alignments and trees to study mammalian evolutionary genomics.
To date, the databases built to gather information on gene orthology do not provide end-users with descriptors of the molecular evolution information and phylogenetic pattern of these orthologues. In this context, we developed OrthoMaM, a database of ORTHOlogous MAmmalian Markers describing the evolutionary dynamics of coding sequences in mammalian genomes. OrthoMaM version 12 includes 15,868 alignments of orthologous coding sequences (CDS) from the 190 complete mammalian genomes currently available. All annotations and 1-to-1 orthology assignments are based on NCBI. Orthologous CDS can be mined for potential informative markers at the different taxonomic levels of the mammalian tree. To this end, several evolutionary descriptors of DNA sequences are provided for querying purposes (e.g. base composition and relative substitution rate). The graphical web interface allows the user to easily browse and sort the results of combined queries. The corresponding multiple sequence alignments and ML trees, inferred using state-of-the art approaches, are available for download both at the nucleotide and amino acid levels. OrthoMaM v12 can be used by researchers interested either in reconstructing the phylogenetic relationships of mammalian taxa or in understanding the evolutionary dynamics of coding sequences in their genomes. OrthoMaM is available for browsing, querying and complete or filtered download at https://orthomam.mbb.cnrs.fr/.


Protective immunity against human carcinoembryonic antigen (CEA) induced by an oral DNA vaccine in CEA-transgenic mice.
Peripheral T-cell tolerance toward human carcinoembryonic self-antigen (CEA) was broken in CEA-transgenic C57BL/6J mice by an oral CEA-based DNA vaccine. This vaccine, delivered by the live, attenuated AroA- strain of Salmonella typhimurium (SL7207), induced tumor-protective immunity mediated by MHC class I-restricted CD8+ T cells. Activation of these T cells was indicated by increased secretion of proinflammatory cytokines IFN-gamma, interleukin (IL)-12 and granulocyte/macrophage-colony stimulating factor, as well as specific tumor rejection and growth suppression in vaccinated CEA-transgenic mice after a lethal challenge with murine MC38 colon carcinoma cells. These tumor cells were double transfected with CEA and the human epithelial cell adhesion molecule (Ep-CAM)/KSA and consequently served as a docking site for a recombinant antibody-IL2 fusion protein (KS1/4-IL2) recognizing KSA. Importantly, the efficacy of the tumor-protective immune response was markedly increased by boosts with this antibody-IL2 fusion protein, resulting in more effective tumor rejection coupled with increased expression of costimulatory molecules B7.2/B7.2 and intercellular adhesion molecule 1 (ICAM-1) on dendritic cells and intensified release of proinflammatory cytokines IFN-gamma, IL-12, and granulocyte/macrophage-colony stimulating factor from T cells of successfully vaccinated CEA-transgenic C57BL/6J mice. Increased T-cell activation mediated by boosts with KS1/4-IL2 fusion protein after tumor cell challenge was further indicated by expanded expression of T-cell activation markers CD25, CD28, CD69, and LFA-1. The application of such CEA-based DNA vaccines and its further improved versions may ultimately prove useful in combination therapies directed against human carcinomas expressing CEA self-antigens.


Psychiatric diagnoses and suicide: revisiting the evidence.
The key role of prevention and treatment of mental disorders in the prevention of suicide is widely acknowledged. Which specific disorders need to be targeted remains to be conclusively demonstrated. To re-examine the presence of psychiatric diagnosis in cases of completed suicide from a global perspective. A review of studies reporting diagnoses of mental disorders in cases of completed suicide with or without history of admission to mental hospitals. Most cases were from Europe and North America (82.2%). The majority (98%) of these had a diagnosis of at least one mental disorder. Among all diagnoses, mood disorders accounted for 30.2%, followed by substance-use related disorders (17.6%), schizophrenia (14.1%), and personality disorders (13.0%). The mental health paradigm in suicide prevention covers just a part of the problem. Antisuicide strategies focusing exclusively on the identification and treatment of depression need to be reconsidered. In addition to this, other mental disorders should be targeted, in particular alcohol-use disorders and schizophrenia. More emphasis should also be placed on psychosocial and environmental interventions diminishing and counteracting stress.


Effector-mediated membrane disruption controls cell death in CBASS antiphage defense.
Cyclic oligonucleotide-based antiphage signaling systems (CBASS) are antiviral defense operons that protect bacteria from phage replication. Here, we discover a widespread class of CBASS transmembrane (TM) effector proteins that respond to antiviral nucleotide signals and limit phage propagation through direct membrane disruption. Crystal structures of the Yersinia TM effector Cap15 reveal a compact 8-stranded β-barrel scaffold that forms a cyclic dinucleotide receptor domain that oligomerizes upon activation. We demonstrate that activated Cap15 relocalizes throughout the cell and specifically induces rupture of the inner membrane. Screening for active effectors, we identify the function of distinct families of CBASS TM effectors and demonstrate that cell death via disruption of inner-membrane integrity is a common mechanism of defense. Our results reveal the function of the most prominent class of effector protein in CBASS immunity and define disruption of the inner membrane as a widespread strategy of abortive infection in bacterial phage defense.


Desmitis of the straight sesamoidean ligament in horses: 9 cases (1995-1997).
To determine clinical signs, diagnostic findings, and outcome for horses with desmitis of the straight sesamoidean ligament (SSL) near its insertion on the middle phalanx. Retrospective study. 9 horses. Medical records were reviewed, and information on signalment, history, clinical signs, diagnostic findings, and treatment was obtained. Follow-up information was obtained through telephone conversations with owners. In all horses, the diagnosis was made by use of high-resolution ultrasonography. Seven horses had moderate lameness on initial examination; lameness was exacerbated in 6 horses following flexion of the distal limb joints. The cause of lameness could not be determined on the basis of clinical signs, and diagnostic local anesthesia was necessary to localize the source of lameness to the distal portion of the limb. Five horses had forelimb involvement (1 bilateral), and 4 had hind limb involvement (1 bilateral). Treatment consisted primarily of a 6-month rest and rehabilitation program. Six of the 9 horses were able to return to their intended use. Results suggest that injury to the SSL proximal to its insertion on the middle phalanx should be considered as a possible cause of lameness in horses, particularly performance horses, with lameness localized to the distal portion of the forelimb or hind limb that do not have any radiographic abnormalities. High-resolution ultrasonography was necessary to make the diagnosis. Horses with an acute injury appeared to have a reasonable chance of responding to treatment and returning to their intended use.


[A presumption calculating formula of the X-ray spectrum generated from a molybdenum target X-ray tube].
A presumption calculating formula of the X-ray spectrum generated from a molybdenum target X-ray tube is presented. The calculation procedure is to add an amount of characteristic X-ray photons that corresponds to the ratio of characteristic photons and bremsstrahlung photons to the bremsstrahlung spectrum obtained using semiempirical calculation. The bremsstrahlung spectrum was calculated by using a corrected Tucker's formula. The corrected content was a formula for calculating the self-absorption length in the target that originated in the difference of the incident angle to the target of the electron and the mass stopping power data. The measured spectrum was separated into the bremsstrahlung component and the characteristic photon component, and the ratio of the characteristic photons and bremsstrahlung photons was obtained. The regression was derived from the function of the tube voltage. Based on this calculation procedure, computer software was constructed that can calculate an X-ray spectrum in arbitrary exposure conditions. The X-ray spectrum obtained from this presumption calculating formula and the measured X-ray spectrum corresponded well. This formula is very useful for analyzing various problems related to mammography by means of Monte Carlo simulations.


Initial size of unilateral pleural effusion determines impact of thoracocentesis on oxygenation.
There have been contradicting reports in the literature regarding the impact of pleural fluid aspiration on patients' oxygenation. The aim of this study was to assess the role of the initial size of effusion on post-drainage oxygenation. We studied 122 patients, aged (mean±SD) 61.2±16.8 years, with unilateral pleural effusion and no remarkable parenchymal lesion, by determining PaO2, PaCO2 and [A-a] PaO2 just before thoracocentesis (T1), 30 min after its completion (T2) and 48 hours after the procedure (T3). Patients were divided into group A (75 patients) with small and moderate sized effusions and group B (47 patients) with large and massive effusions. The position of the meniscus line on the posteroanterior film, being arbitrarily set at just above the upper costal margin of the sixth anterior rib, was used to divide the two groups. Patients were studied at rest, breathing room air in the sitting position. Repeated measures ANOVA (related samples) and the Friedman test when the normality assumption was violated were used. In group A, at T3, PaO2(mm Hg) showed a statistically significant increase versus T1 (p<0.001) and T2 (p=0.002), while [A-a] PaO2 displayed a statistically significant decrease compared with T1 (p<0.001) and T2 (p=0.001). In group B, at T2, PaO2 presented significant decrease versus T1 (p<0.001) and T3 (p<0.001), while [A-a] PO2 was found to be significantly increased compared with both T1 and T3 (p<0.001). Patients with smaller effusions showed a small improvement in their oxygenation 48 hours post-thoracocentesis (T3). Patients with larger effusions exhibited a transient reduction in their oxygenation immediately after fluid removal (T2).


Shining light on the microscopic resonant mechanism responsible for cavity-mediated chemical reactivity.
Strong light-matter interaction in cavity environments is emerging as a promising approach to control chemical reactions in a non-intrusive and efficient manner. The underlying mechanism that distinguishes between steering, accelerating, or decelerating a chemical reaction has, however, remained unclear, hampering progress in this frontier area of research. We leverage quantum-electrodynamical density-functional theory to unveil the microscopic mechanism behind the experimentally observed reduced reaction rate under cavity induced resonant vibrational strong light-matter coupling. We observe multiple resonances and obtain the thus far theoretically elusive but experimentally critical resonant feature for a single strongly coupled molecule undergoing the reaction. While we describe only a single mode and do not explicitly account for collective coupling or intermolecular interactions, the qualitative agreement with experimental measurements suggests that our conclusions can be largely abstracted towards the experimental realization. Specifically, we find that the cavity mode acts as mediator between different vibrational modes. In effect, vibrational energy localized in single bonds that are critical for the reaction is redistributed differently which ultimately inhibits the reaction.


Inhibition of scavenger receptor-mediated modified low-density lipoprotein endocytosis in cultured bovine aortic endothelial cells by the glycoprotein processing inhibitor castanospermine.
Castanospermine (1,6,7,8-tetrahydroxyoctahydroindolizidine) is a plant alkaloid that inhibits alpha-glucosidases, including the glycoprotein processing glucosidase I. When endothelial cells were grown for 48 h, or longer, in the presence of this alkaloid, they produced scavenger receptors for modified low-density lipoproteins (LDL) that had mostly Glc3Man7-9(GlcNAc)2 structures rather than the usual complex types of oligosaccharides. Furthermore, growth in the presence of castanospermine resulted in a substantial inhibition in degradation of endocytosed 125I-acetylated LDL, as well as a dose-dependent inhibition of 125I-acetylated LDL binding to these cells. Scatchard analysis of binding curves indicated that the diminished binding was due to a decrease in the number of scavenger receptor molecules at the cell surface rather than to a change in the affinity of the receptors for their ligand. Since castanospermine-treated cells had the same total number of cellular receptor molecules as did controls cells, it seemed likely that castanospermine caused an alteration in receptor targeting, rather than an inhibition in receptor synthesis or a stimulation in receptor degradation. Density gradient fractionation of cell homogenates showed that castanospermine-treated cells did have a much greater percentage of scavenger LDL receptor molecules in the endoplasmic reticulum-Golgi fraction and fewer receptors in the plasma membrane fraction, whereas normal cells showed the opposite distribution.


The soybean sucrose binding protein gene family: genomic organization, gene copy number and tissue-specific expression of the SBP2 promoter.
The sucrose binding protein (SBP) from soybean has been implicated as an important component of the sucrose uptake system. Two SBP genomic clones, gsS641.1 and gsS641.2, which correspond to allelic forms of the GmSBP2/S64 gene, have been isolated and characterized. As a member of the seed storage protein superfamily, it has been shown that the SBP gene structure is similar to vicilin genes with intron/exon boundaries at conserved positions. Fluores cence in situ hybridization (FISH) suggested that the soybean SBP gene family is represented by at least two non-allelic genes corresponding to the previously isolated GmSBP1 and GmSBP2/S64 cDNAs. These two cDNAs share extensive sequence similarity but are located at different loci in the soybean genome. To investigate transcriptional activation of the GmSBP2 gene, 2 kb 5'-flanking sequences of gsS641.1 and gsS641.2 were fused to the beta-glucuronidase (GUS) reporter gene and to the green fluorescent protein (GFP) reporter gene and inde pendently introduced into Nicotiana tabacum by Agrobacterium tumefaciens-mediated transformation. The SBP2 promoter directed expression of both GUS and GFP reporter genes with high specificity to the phloem of leaves, stems and roots. Thus, the overall pattern of SBP-GUS or SBP-GFP expression is consistent with the involvement of SBP in sucrose translocation-dependent physiological processes.


Analysis of the seismic performance of isolated buildings according to life-cycle cost.
This paper proposes an indicator of seismic performance based on life-cycle cost of a building. It is expressed as a ratio of lifetime damage loss to life-cycle cost and determines the seismic performance of isolated buildings. Major factors are considered, including uncertainty in hazard demand and structural capacity, initial costs, and expected loss during earthquakes. Thus, a high indicator value indicates poor building seismic performance. Moreover, random vibration analysis is conducted to measure structural reliability and evaluate the expected loss and life-cycle cost of isolated buildings. The expected loss of an actual, seven-story isolated hospital building is only 37% of that of a fixed-base building. Furthermore, the indicator of the structural seismic performance of the isolated building is much lower in value than that of the structural seismic performance of the fixed-base building. Therefore, isolated buildings are safer and less risky than fixed-base buildings. The indicator based on life-cycle cost assists owners and engineers in making investment decisions in consideration of structural design, construction, and expected loss. It also helps optimize the balance between building reliability and building investment.


Determinants of bluetongue virus virulence in murine models of disease.
Bluetongue is a major infectious disease of ruminants that is caused by bluetongue virus (BTV). In this study, we analyzed virulence and genetic differences of (i) three BTV field strains from Italy maintained at either a low (L strains) or high (H strains) passage number in cell culture and (ii) three South African "reference" wild-type strains and their corresponding live attenuated vaccine strains. The Italian BTV L strains, in general, were lethal for both newborn NIH-Swiss mice inoculated intracerebrally and adult type I interferon receptor-deficient (IFNAR(-/-)) mice, while the virulence of the H strains was attenuated significantly in both experimental models. Similarly, the South African vaccine strains were not pathogenic for IFNAR(-/-) mice, while the corresponding wild-type strains were virulent. Thus, attenuation of the virulence of the BTV strains used in this study is not mediated by the presence of an intact interferon system. No clear distinction in virulence was observed for the South African BTV strains in newborn NIH-Swiss mice. Full genomic sequencing revealed relatively few amino acid substitutions, scattered in several different viral proteins, for the strains found to be attenuated in mice compared to the pathogenic related strains. However, only the genome segments encoding VP1, VP2, and NS2 consistently showed nonsynonymous changes between all virulent and attenuated strain pairs. This study established an experimental platform for investigating the determinants of BTV virulence. Future studies using reverse genetics will allow researchers to precisely map and "weight" the relative influences of the various genome segments and viral proteins on BTV virulence.


The application of tDCS for the treatment of psychiatric diseases.
Neuroplasticity represents the dynamic structural and functional reorganization of the central nervous system, including its connectivity, due to environmental and internal demands. It is recognized as a major physiological basis for adaption of cognition and behaviour, and, thus, of utmost importance for normal brain function. Cognitive dysfunctions are major symptoms in psychiatric disorders, which are often associated with pathological alteration of neuroplasticity. Transcranial direct current stimulation (tDCS), a recently developed non-invasive brain stimulation technique, is able to induce and modulate cortical plasticity in humans via the application of relatively weak current through the scalp of the head. It has the potential to alter pathological plasticity and restore dysfunctional cognitions in psychiatric diseases. In the last decades, its efficacy to treat psychiatric disorders has been explored increasingly. This review will give an overview of pathological alterations of plasticity in psychiatric diseases, gather clinical studies involving tDCS to ameliorate symptoms, and discuss future directions of application, with an emphasis on optimizing stimulation effects.


The Sport Concussion Assessment Tool 5th Edition (SCAT5): Background and rationale.
This paper presents the Sport Concussion Assessment Tool 5th Edition (SCAT5), which is the most recent revision of a sport concussion evaluation tool for use by healthcare professionals in the acute evaluation of suspected concussion. The revision of the SCAT3 (first published in 2013) culminated in the SCAT5. The revision was based on a systematic review and synthesis of current research, public input and expert panel review as part of the 5th International Consensus Conference on Concussion in Sport held in Berlin in 2016. The SCAT5 is intended for use in those who are 13 years of age or older. The Child SCAT5 is a tool for those aged 5-12 years, which is discussed elsewhere.


The effects of the endocrine disruptors dithiocarbamates on the mammalian ovary with particular regard to mancozeb.
Many human-made chemicals are called endocrine disruptors (EDs) because they have the potential to disrupt endocrine functions in exposed organisms. Many EDs can disrupt hormonal homeostasis by interfering with hormone receptor recognition, binding and activation, while others act by still unknown mechanisms. Among the EDs specifically affecting the female reproductive system, those with steroidogenic/antisteroidogenic effects have been extensively studied and the mechanisms of toxicity clarified also at molecular level. For many others, information is restricted to few epidemiological data and in vivo/in vitro experiments with animal models. This is the case of the dithiocarbamates, and in particular of the fungicide mancozeb, an ethylenedithiocarbamate widely used to protect fruit and vegetables, ginseng included, because of its low acute toxicity in humans. Although the mechanism(s) by which mancozeb may specifically act on female reproductive organs are largely unknown, data on experimental animals in vivo have demonstrated that the fungicide can induce several disturbances on estrus cycle. When used in vitro at concentrations considered too low to cause human health injuries, the fungicide impairs mouse embryo development and meiotic spindle assembly. The possibility that the female germ cell (the oocyte) could be a specific target of mancozeb suggests a role for this fungicide as probable inductor of infertility also in exposed human populations.


[Intralimbic evoked potentials in the formation of an instrumental food conditioned reflex to hippocampal electrostimulation in dogs].
In dogs with the electrodes implanted in the hippocampus, amygdala, septum and hypothalamus an instrumental alimentary conditioned reflex (CR) was elaborated to electrostimulation of the hippocampus. Intralimbic evoked potentials (EPs) were studied during the elaboration and extinction of this reflex and during stimulations of limbic structures conducted with the purpose of CR generalization checking. Late EP components in the lateral hypothalamus and central nucleus of the amygdala changed during CR elaboration and extinction and in the hippocampus during amygdala testing. In both cases the amplitude of trace positivity and of slow negative wave was less, when during stimulation of the structure an instrumental movement was initiated than at its absence.


Effects of oil-soluble organosulfur compounds from garlic on doxorubicin-induced lipid peroxidation.
Clinical efficacy of doxorubicin is compromised due to free radical generation leading to cardiac toxicity. Oil-soluble organosulfur compounds, diallyl sulfide (DAS), diallyl disulfide (DADS), dipropyl sulfide (DPS) and dipropyl disulfide (DPDS), present in garlic were examined for their antiperoxidant effects. DADS inhibited liver microsomal lipid peroxidation induced by NADPH, ascorbate and doxorubicin. DAS, DPS and DPDS were ineffective inhibitors of liver microsomal lipid peroxidation. DADS could be used in combination with doxorubicin to protect oxidative injuries to improve the clinical efficacy of doxorubicin.


ICU admissions from an obstetrical hospital.
To ascertain the anaesthetic complications requiring post-anaesthetic respiratory support in a large obstetrical hospital, the hospital records of obstetrical patients admitted to an adjacent general intensive care unit (ICU) were studied. Obstetrical patients who required mechanical ventilation following anaesthetic complications were identified and their hospital records reviewed. In a ten-year period there were 61,435 women delivered at the Mater Mothers Hospital (MMH) in Brisbane, Queensland, Australia. Of these 24,161 had an anaesthetic associated with delivery including 7,452 general anaesthetics (GAs) and 16,709 regional anaesthetics (RAs). There were also 3,708 GAs and 17 RAs for procedures other than delivery. During this period there were 126 obstetrical admissions to the ICU. Sixteen were due to anaesthetic complications, twelve followed GA and four followed RA. Complications included anaphylaxis, high block and failure of tracheal intubation. The incidence of a major complication of a GA causing admission to the ICU was 1 in 932 and for RA was 1 in 4177 when these were given for delivery (P less than 0.01). If a complication requiring ICU admission and mechanical ventilation is used as the criterion of safety it appears that RA is safer than a GA for delivery.


Viral factors required for interferon induction by Newcastle disease virus in mouse macrophages and chicken embryo cells.
The triggering mechanism for interferon synthesis in mouse peritoneal macrophages and chick embryo (CE) cells by Newcastle disease virus (NDV) exposed to hydroxylamine or homologous antiserum was investigated in relation to the intracellular fate of these agents. Inactivation of NDV at 22 degrees C by I M-hydroxylamine proceeded with first-order kinetics, whereas the interferon-inducing capacity of hydroxylamine-treated virus in macrophages was unimpaired. In contrast to infective NDV, hydroxylamine-inactivated virus produced interferon in CE cells, and such a virus still had partial RNA-dependent RNA polymerase activity. Hydroxylamine-inactivated NDV was adsorbed to and uncoated in both normal and chloroquine diphosphate treated cells, but no viral double-stranded RNA was detected. Hydroxylamine treatment of virion-extracted RNA and neutralization of intact virions by antibody abolished the capacity of the virus to induce interferon. Infective as well as neutralized NDV interacted with macrophages to the same degree, but association between NDV and CE cells was prevented by antibody-coating. In macrophages, the RNA of neutralized NDV became more sensitive to RNase than RNA of infective NDV, but this process was inhibited in chloroquine diphosphate-treated cells. These results suggest that interferon induction by NDV involves components of the virion which are present up to the regular uncoating process.


Studies on the substrate-binding sites of liver microsomal cytochrome P-448.
The interaction of substrates of the microsomal mixed-function oxidases with cytochromes P-450 and P-448 was investigated by using liver microsomes from rats pretreated with phenobarbital or 3-methylcholanthrene, and with purified forms of the cytochromes isolated from rabbit liver. The two forms of the cytochrome have different substrate specificities; cytochrome P-450 has one type 1 substrate-binding site that can accommodate a large variety of substrates, but in contrast cytochrome P-448 may possess two type 1 substrate-binding sites, one of which is different to that of cytochrome P-450 in that it shows a specificity for substrates such as safrole and 9-hydroxy-ellipticine. These findings explain why the two forms of the cytochrome have different substrate specificities and play contrasting roles in the activation and deactivation of xenobiotics.


Environmental factors influencing spotted hyena and lion population biomass across Africa.
The spotted hyena (Crocuta crocuta Erxleben) and the lion (Panthera leo Linnaeus) are two of the most abundant and charismatic large mammalian carnivores in Africa and yet both are experiencing declining populations and significant pressures from environmental change. However, with few exceptions, most studies have focused on influences upon spotted hyena and lion populations within individual sites, rather than synthesizing data from multiple locations. This has impeded the identification of over-arching trends behind the changing biomass of these large predators. Using partial least squares regression models, influences upon population biomass were therefore investigated, focusing upon prey biomass, temperature, precipitation, and vegetation cover. Additionally, as both species are in competition with one other for food, the influence of competition and evidence of environmental partitioning were assessed. Our results indicate that spotted hyena biomass is more strongly influenced by environmental conditions than lion, with larger hyena populations in areas with warmer winters, cooler summers, less drought, and more semi-open vegetation cover. Competition was found to have a negligible influence upon spotted hyena and lion populations, and environmental partitioning is suggested, with spotted hyena population biomass greater in areas with more semi-open vegetation cover. Moreover, spotted hyena is most heavily influenced by the availability of medium-sized prey biomass, whereas lion is influenced more by large size prey biomass. Given the influences identified upon spotted hyena populations in particular, the results of this study could be used to highlight populations potentially at greatest risk of decline, such as in areas with warming summers and increasingly arid conditions.


Identification and structural analysis of dimeric chicken complement component 3d and its binding with chicken complement receptor 2.
Complement component 3d (C3d), the final cleavage product of complement component C3, interacts with CR2 and thus plays a crucial role in linking the innate and adaptive immune systems. Additionally, human C3d executes various functions in its dimeric form, which is more effective than its monomeric form. In this study, we aimed to explored whether chicken C3d (chC3d) exhibits similar characteristics, namely dimerization and binding of dimeric chC3d to chicken CR2 (chCR2). We investigated the interaction and co-localization of chC3d with itself using coimmunoprecipitation and confocal laser scanning microscopy, respectively. Then, dimeric chC3d was detected using native polyacrylamide gel electrophoresis and western blotting, and its equilibrium dissociation constant KD (827 nM) was determined using surface plasmon resonance. Finally, the interaction modes of dimeric chC3d were identified using molecular docking simulations, which revealed that dimeric chC3d could crosslink with chCR2 receptor. Overall, our findings will facilitate future explorations of the chicken complement system.


Patient-controlled encrypted genomic data: an approach to advance clinical genomics.
The revolution in DNA sequencing technologies over the past decade has made it feasible to sequence an individual's whole genome at a relatively low cost. The potential value of the information generated by genomic technologies for medicine and society is enormous. However, in order for exome sequencing, and eventually whole genome sequencing, to be implemented clinically, a number of major challenges need to be overcome. For instance, obtaining meaningful informed-consent, managing incidental findings and the great volume of data generated (including multiple findings with uncertain clinical significance), re-interpreting the genomic data and providing additional counselling to patients as genetic knowledge evolves are issues that need to be addressed. It appears that medical genetics is shifting from the present "phenotype-first" medical model to a "data-first" model which leads to multiple complexities. This manuscript discusses the different challenges associated with integrating genomic technologies into clinical practice and describes a "phenotype-first" approach, namely, "Individualized Mutation-weighed Phenotype Search", and its benefits. The proposed approach allows for a more efficient prioritization of the genes to be tested in a clinical lab based on both the patient's phenotype and his/her entire genomic data. It simplifies "informed-consent" for clinical use of genomic technologies and helps to protect the patient's autonomy and privacy. Overall, this approach could potentially render widespread use of genomic technologies, in the immediate future, practical, ethical and clinically useful. The "Individualized Mutation-weighed Phenotype Search" approach allows for an incremental integration of genomic technologies into clinical practice. It ensures that we do not over-medicalize genomic data but, rather, continue our current medical model which is based on serving the patient's concerns. Service should not be solely driven by technology but rather by the medical needs and the extent to which a technology can be safely and effectively utilized.


[Approach to the development of an actovaccine using vaccinia virus as a model].
Injection of living vaccinia virus to the mesenteric vein of rabbits leads to its accumulation in liver in high concentration. In the other series of experiments vaccinia virus inactivated with gamma-radiation was inoculated into the mesenteric vein. Later on the animals were sacrificed at various dates. Homogenates were prepared from their liver. Comparative study of these homogenates made it possible to reveal increased immunogenic activity of homogenate obtained 24 hours after intravenous inoculation of inactivated vaccinia virus into animals. Immunogenic activity of this homogenate was more clearly manifested than immunogenic activity of the antigen itself--inactivated vaccinia vorus. Neither original inactivated vaccinia virus nor inactivated virus absorbed by Kupffer cells induced antibody formation. It is supposed that vaccines may be prepared from various viruses or tumor cells treated with macrophages with subsequent chemical extraction of the most immunogenic fraction.


Toward the prevention of childhood undernutrition: diet diversity strategies using locally produced food can overcome gaps in nutrient supply.
Chronic undernutrition is prevalent in Mozambique, where children suffer from stunting, vitamin A deficiency, anemia, and other nutrition-related disorders. Complete diet formulation products (CDFPs) are increasingly promoted to prevent chronic undernutrition. Using linear programming, to investigate whether diet diversification using local foods should be prioritized in order to reduce the prevalence of chronic undernutrition. Market prices of local foods were collected in Tete City, Mozambique. Linear programming was applied to calculate the cheapest possible fully nutritious food baskets (FNFB) by stepwise addition of micronutrient-dense localfoods. Only the top quintile of Mozambican households, using average expenditure data, could afford the FNFB that was designed using linear programming from a spectrum of local standard foods. The addition of beef heart or liver, dried fish and fresh moringa leaves, before applying linear programming decreased the price by a factor of up to 2.6. As a result, the top three quintiles could afford the FNFB optimized using both diversification strategy and linear programming. CDFPs, when added to the baskets, were unable to overcome the micronutrient gaps without greatly exceeding recommended energy intakes, due to their high ratio of energy to micronutrient density. Dietary diversification strategies using local, low-cost, nutrient-dense foods can meet all micronutrient recommendations and overcome all micronutrient gaps. The success of linear programming to identify a low-cost FNFB depends entirely on the investigators' ability to select appropriate micronutrient-dense foods. CDFPs added to food baskets are unable to overcome micronutrient gaps without greatly exceeding recommended energy intake.


Kidney Response to Heart Failure: Proteomic Analysis of Cardiorenal Syndrome.
Chronic heart failure (HF) disrupts normal kidney function and leads to cardiorenal syndrome that further promotes HF progression. To identify potential participants in HF-related injury, we analyzed kidney proteome in an established HF model. HF was induced by chronic volume overload in male HanSD rats using aorto-caval fistula. After 21 weeks, cardiac and renal functions (in-situ kidney study) and renal proteomics were studied in sham-operated (controls) and HF rats, using iTRAQ labeling and LC-MS with Orbitrap Fusion, leading to identification and quantification of almost 4000 proteins. Compared to controls, HF rats had cardiac hypertrophy, systemic and pulmonary congestion. Kidneys of HF rats had reduced renal blood flow, sodium excretion and urine production. While glomerular filtration rate, serum cystatin C and creatinine were still normal compared to controls, HF kidneys showed albuminuria and markedly increased tissue angiotensin-II levels (5-fold). HF kidneys (versus controls) displayed differential expression (˃1.5-fold) of 67 proteins. The most upregulated were angiotensin-converting enzyme (ACE, ˃20-fold), advanced glycosylation product-specific receptor (RAGE, 14-fold), periostin (6.8-fold), caveolin-1 (4.5-fold) and other proteins implicated in endothelial function (vWF, cavins 1-3, T-kininogen 2), proinflammatory ECM activation (MFAP4, collagen-VI, galectin-3, FHL-1, calponin) and proteins involved in glomerular filtration membrane integrity (CLIC5, ZO-1). Carboxylesterase-1D (CES1D), an enzyme that converts ACE inhibitors or sacubitril into active drugs, was also upregulated in HF kidneys. Chronic HF leads to latent kidney injury, associated with deep changes in kidney protein composition. These alterations may act in concert with intrarenal renin-angiotensin system activation and may serve as markers and/or targets to tackle cardiorenal syndrome.


Porphyria cutanea tarda and systemic lupus erythematosus.
Fifty-five patients with porphyria cutanea tarda (PCT) were examined for the simultaneous occurrence of systemic lupus erythematosus (SLE). The diagnostic criteria for SLE were those set up by the American Rheumatism Association (ARA). No patient fulfilled the ARA criteria for SLE. Clinical and laboratory features characteristically associated with SLE (although they were not ARA criteria) were simultaneously evaluated. A total of 120 items occurred in those cases but only 15 of these were ARA criteria. These findings, which may suggest the presence of SLE in patients with PCT, might be ascribed to the concomitant liver disease. The simultaneous occurrence of SLE and PCT is reviewed from the literature. The ARA criteria were fulfilled in one third of the reported cases. The coexistence of SLE and PCT might be less frequent than formerly believed.


Deciphering the connectivity structure of biological networks using MixNet.
As biological networks often show complex topological features, mathematical methods are required to extract meaningful information. Clustering methods are useful in this setting, as they allow the summary of the network's topology into a small number of relevant classes. Different strategies are possible for clustering, and in this article we focus on a model-based strategy that aims at clustering nodes based on their connectivity profiles. We present MixNet, the first publicly available computer software that analyzes biological networks using mixture models. We apply this method to various networks such as the E. coli transcriptional regulatory network, the macaque cortex network, a foodweb network and the Buchnera aphidicola metabolic network. This method is also compared with other approaches such as module identification or hierarchical clustering. We show how MixNet can be used to extract meaningful biological information, and to give a summary of the networks topology that highlights important biological features. This approach is powerful as MixNet is adaptive to the network under study, and finds structural information without any a priori on the structure that is investigated. This makes MixNet a very powerful tool to summarize and decipher the connectivity structure of biological networks.


Bone mineral content and body size 65 to 100 weeks' postconception in preterm and full term infants.
A total of 36 preterm and 22 full term infants were weighed and measured at 65 to 100 weeks' postconception. It was found that the preterm infants were on average significantly lighter by 1008 g and shorter by 3.8 cm than those born at full term. Despite the smaller size of preterm infants, mean values of bone mineral content in the mid-forearm were not significantly different between the two groups. In contrast, near 40 weeks' postconception the mean bone mineral content observed in 35 of the preterm infants was significantly smaller than that observed in eight of the full term infants. Our results suggest that there is a phase of rapid mineral accretion between 40 and 60 weeks' postconception. This 'catch up' in mineral accretion reduces the perinatal mineralisation deficit that might otherwise persist into childhood.


Misclassification in administrative claims data: quantifying the impact on treatment effect estimates.
Misclassification is present in nearly every epidemiologic study, yet is rarely quantified in analysis in favor of a focus on random error. In this review, we discuss past and present wisdom on misclassification and what measures should be taken to quantify this influential bias, with a focus on bias in pharmacoepidemiologic studies. To date, pharmacoepidemiology primarily utilizes data obtained from administrative claims, a rich source of prescription data but susceptible to bias from unobservable factors including medication sample use, medications filled but not taken, health conditions that are not reported in the administrative billing data, and inadequate capture of confounders. Due to the increasing focus on comparative effectiveness research, we provide a discussion of misclassification in the context of an active comparator, including a demonstration of treatment effects biased away from the null in the presence of nondifferential misclassification. Finally, we highlight recently developed methods to quantify bias and offer these methods as potential options for strengthening the validity and quantifying uncertainty of results obtained from pharmacoepidemiologic research.


Protein kinase substrate identification on functional protein arrays.
Over the last decade, kinases have emerged as attractive therapeutic targets for a number of different diseases, and numerous high throughput screening efforts in the pharmaceutical community are directed towards discovery of compounds that regulate kinase function. The emerging utility of systems biology approaches has necessitated the development of multiplex tools suitable for proteomic-scale experiments to replace lower throughput technologies such as mass spectroscopy for the study of protein phosphorylation. Recently, a new approach for identifying substrates of protein kinases has applied the miniaturized format of functional protein arrays to characterize phosphorylation for thousands of candidate protein substrates in a single experiment. This method involves the addition of protein kinases in solution to arrays of immobilized proteins to identify substrates using highly sensitive radioactive detection and hit identification algorithms. To date, the factors required for optimal performance of protein array-based kinase substrate identification have not been described. In the current study, we have carried out a detailed characterization of the protein array-based method for kinase substrate identification, including an examination of the effects of time, buffer compositions, and protein concentration on the results. The protein array approach was compared to standard solution-based assays for assessing substrate phosphorylation, and a correlation of greater than 80% was observed. The results presented here demonstrate how novel substrates for protein kinases can be quickly identified from arrays containing thousands of human proteins to provide new clues to protein kinase function. In addition, a pooling-deconvolution strategy was developed and applied that enhances characterization of specific kinase-substrate relationships and decreases reagent consumption. Functional protein microarrays are an important new tool that enables multiplex analysis of protein phosphorylation, and thus can be utilized to identify novel kinase substrates. Integrating this technology with a systems biology approach to cell signalling will help uncover new layers in our understanding of this essential class of enzymes.


Role of JNK activation in paclitaxel-induced apoptosis in human head and neck squamous cell carcinoma.
It has been reported that paclitaxel activates cell cycle arrest and increases caspase protein expression to induce apoptosis in head and neck squamous cell carcinoma (HNSCC) cell lines. However, the potential signaling pathway regulating this apoptotic phenomenon remains unclear. The present study used OEC-M1 cells to investigate the underlying molecular mechanism of paclitaxel-induced apoptosis. Following treatment with paclitaxel, cell viability was assessed via the MTT assay. Necrosis, apoptosis, cell cycle and mitochondrial membrane potential (∆Ψm) were analyzed via flow cytometric analyses, respectively. Western blot analysis was performed to detect the expression levels of proteins associated with the MAPK and caspase signaling pathways. The results demonstrated that low-dose paclitaxel (50 nM) induced apoptosis but not necrosis in HNSCC cells. In addition, paclitaxel activated the c-Jun N-terminal kinase (JNK), but not extracellular signal-regulated kinase or p38 mitogen-activated protein kinase. The paclitaxel-activated JNK contributed to paclitaxel-induced apoptosis, activation of caspase-3, -6, -7, -8 and -9, and reduction of ∆Ψm. In addition, caspase-8 and -9 inhibitors, respectively, significantly decreased paclitaxel-induced apoptosis. Notably, Bid was truncated following treatment with paclitaxel. Taken together, the results of the present study suggest that paclitaxel-activated JNK is required for caspase activation and loss of ∆Ψm, which results in apoptosis of HNSCC cells. These results may provide mechanistic basis for designing more effective paclitaxel-combining regimens to treat HNSCC.


Validation and determination of the sensing area of the KINOTEX sensor as part of development of a new mattress with an interface pressure-sensing system.
The purpose of the present study was to examine the validity of the KINOTEX sensor via comparison with an existing sensor and to determine the sensing areas for a new alternating-air mattress that incorporates an interface pressure-sensing system. The study design was an evaluation study to validate and determine the sensing area of the KINOTEX sensor in comparison with another sensor. Study participants were fifty-one healthy volunteers over eighteen years of age, and the two sensors were placed between participants and an alternating-air mattress. We measured the contact area, full weight load, and maximum pressure in the calcaneal region using two sensors and obtained a graphic pressure distribution of >40 mmHg in the lateral and supine positions. Correlation coefficients between sensors were r = 0.88 (p < 0.001) for the contact area, r = 0.89 (p < 0.001) for full weight load, and r = 0.72 (p < 0.001) at maximum pressure in the calcaneal region. Ninety-one percent of the pressure distribution was recorded in the central 50 cm of the bed, and 94.6% was recorded within an area 160 cm in length need the foot of the bed. We investigated the correlation between the KINOTEX sensor and an existing sensor and determined the necessary sensing area. Results suggested the feasibility of developing a new alternating-air mattress incorporating an interface pressure-sensing system to help prevent pressure ulcers.


New treatment paradigms in spondyloarthritis.
The review presents the recent rapid expansion of therapeutical options in spondyloarthritis. Additionally, it focuses on the importance of additional questions raised by the growing therapeutic possibilities related to the optimal use of these drugs. The emergence of new treatment options opens new avenues and opportunities for treating patients with nonresponse, contraindications, or intolerance for classic drugs. However, it becomes more relevant than ever to define not only drugs and treatment options but also treatment strategies. We address current literature and remaining questions on strategies such as early intervention, combination treatment, personalized medicine, and treat-to-target. Not only the treatment as such, but also the treatment strategy is crucial to reveal the full therapeutic potential and benefit for patients. Whereas cautious but crucial steps have been taken in the last years to explore these aspects, related to timing and sequence of treatment (including combination treatments), stratified medicine approaches, and treat-to-target strategies, it is now time for full-scale investment in prospective strategy trials.


Validation of the NeoFilm for Yeast and Mold Method for Enumeration of Yeasts and Molds in Select Foods.
NeoFilm Yeast and Mold (Y&M), also known as Sanita-kun Yeasts and Molds, is a simple, effective device used for the enumeration of yeasts and molds. It consists of a nonwoven fabric on which a layer of microbial nutrients is deposited in a film. A 1 mL sample homogenate is applied to the membrane and this, in turn, is incubated for 48-72 h at 25°C. Sample homogenates were prepared using two different diluents for customer convenience: phosphate buffered saline (PBS) and 0.1% peptone water. In comparative testing of breaded chicken nuggets, dry pet food, orange juice concentrate, yogurt, and cake mix, there were statistically significant differences in the counts obtained by the NeoFilm Y&M and U.S. Food and Drug Administration Bacteriological Analytical Manual reference culture methods only in the following instances: medium level for orange juice with PBS as diluent and low level for pet food with 0.1% peptone water as diluent, where reference method counts were higher than those of NeoFilm; medium level for cake mix with PBS, and low and medium levels for cake mix with 0.1% peptone water, where NeoFilm produced higher counts than the reference method. In addition to the method comparison study with five matrixes, robustness and stability/lot-to-lot testing were also performed. Results of robustness testing showed no significant effect on results even with perturbation to three assay parameters simultaneously. Results of testing of three lots of devices ranging in age from 2 to 26 months post-manufacture showed no significant differences in performance.


Acidithiobacillus species drive the formation of ferric-silica cemented microstructure: Insights into early hardpan development for mine site rehabilitation.
Hardpan-based profiles naturally formed under semi-arid climatic conditions have substantial potential in rehabilitating sulfidic tailings, resulting from their aggregation microstructure regulated by Fe-Si cements. Nevertheless, eco-engineered approaches for accelerating the formation of complex cementation structure remain unclear. The present study aims to investigate the microbial functions of extremophiles on mineral dissolution, oxidation, and aggregation (cementation) through a microcosm experiment containing pyrites and polysilicates, of which are dominant components in typical sulfidic tailings. Microspectroscopic analysis revealed that pyrite was rapidly dissolved and massive microbial corrosion pits were displayed on pyrite surfaces. Synchrotron-based X-ray absorption spectroscopy demonstrated that approximately 30 % pyrites were oxidized to jarosite-like (ca. 14 %) and ferrihydrite-like minerals (ca. 16 %) in talc group, leading to the formation of secondary Fe precipitates. The Si ions co-dissolved from polysilicates may be embedded into secondary Fe precipitates, while these clustered Fe-Si precipitates displayed distinct morphology (e.g., "circular" shaped in the talc group, "fine-grained" shaped in the chlorite group, and "donut" shaped in the muscovite group). Moreover, the precipitates could join together and act as cementing agents aggregating mineral particles together, forming macroaggregates in talc and chlorite groups. The present findings revealed critical microbial functions on accelerating mineral dissolution, oxidation, and aggregation of pyrite and various silicates, which provided the eco-engineered feasibility of hardpan-based technology for mine site rehabilitation.


Preparation and purification of microplasmin.
A catalytically active, human microplasmin was produced by incubation of [Lys]plasmin in buffer at pH 11.0 for up to 12 hr. The microplasmin was purified by affinity chromatography that used lysine-Sepharose and soybean trypsin inhibitor-Sepharose columns. It is homogeneous and pure by electrophoretic analysis in NaDodSO4/polyacrylamide gels and by gel filtration on a Superose 12 column. The molecular weight of the microplasmin determined by NaDodSO4 gel electrophoresis is 29,000 and 26,500 under reducing condition, whereas the molecular weight of native plasmin is 76,500. Microplasmin consists mainly of the ligh (B) chain of native human plasmin and possesses one active site per protein molecule when titrated with p-nitrophenyl p'-guanidinobenzoate. Microplasmin hydrolyzes the peptide substrate NH2-D-Val-Leu-Lys-p-nitroanilide (S-2251) with a Km of 0.361 +/- 0.017 mM and a kcat of 40.3 +/- 3.3 s-1 at pH 7.4 and 37 degrees C, whereas native plasmin has a Km of 0.355 +/- 0.002 mM and a kcat of 27.9 +/- 0.3 s-1 under the same conditions.


Genomic data in the electronic medical record: perspectives from a biobank community advisory board.
A proof of principle pharmacogenomic translational study was used as a case example to explore Biobank Community Advisory Board (CAB) member views about placing genomic information into the medical record and to establish how CAB input could affect research design. CAB members expressed enthusiasm for the potential benefit of the research discussed, yet voiced concerns regarding the recruitment and consent materials. They discussed the value of genomic research and its clinical utility; the risk of genetic discrimination; and personal ownership of genomic data. Members distinguished between indirect benefits to future generations and individual risk to research participants. Feedback was used to revise the recruitment and consent materials. Results highlight tensions reported between the public's support for genomic research and concerns with genomic information in the medical record and its use in medical decision-making.


Powdered milk micellar casein prevents oral colonization by Streptococcus sobrinus and dental caries in rats: a basis for the caries-protective effect of dairy products.
Three animal studies were performed to investigate the influence of the macromolecular structure of milk casein on caries incidence and the possible ecological changes of the oral microbiota by such casein fractions. Towards this end, rats were infected with mixed bacterial suspensions of Streptococcus sobrinus OMZ 176 and Actinomyces viscosus Ny1. Various milk protein fractions were incorporated into carefully balanced powdered cariogenic diets to constitute the sole major protein component. Diets containing micellar casein had a pronounced and highly significant effect on almost all clinical and microbiological parameters examined. Both the formation of advanced dentinal fissure (B) and smooth surface (E) caries lesions was inhibited by diets containing micellar casein; this caries-inhibiting effect appeared to be due mainly to modifications within the plaque microbiota. The proportion of S. sobrinus in the oral cavity of rats was reduced (73-80%) by micellar casein-containing preparations, whereas the A. viscosus population was increased. Both these microbiological parameters were always negatively correlated. This appears to be the first example of a food component other than dietary sugars, selectively modifying the composition of the dental plaque microbiota of rats in such a way as to reduce its pathogenic potential. It also demonstrates the importance of establishing a molecular basis for the role of food components, which prove to be beneficial to oral health.


The impact of COVID-19 pandemic on gastrointestinal diseases: a single-center cross-sectional study in central Greece.
The current COVID-19 pandemic induced a suppressive environment for healthcare professionals and patients, especially during the lockdown period. Except for the direct burden of the COVID-19, collateral damage has been identified concerning other diseases. The aim of this study was to evaluate the potential impact of the lockdown on the non-COVID-19 patients' outcome in a tertiary gastroenterology department. Patients admitted to our department during the lockdown period (23 March- 4 May 2020) and during the respective previous year's timeframe were recruited. Sex, age, comorbidities, presenting symptoms, final diagnosis, therapeutic management, duration of hospitalization, and outcome were evaluated. A direct comparison was performed to investigate the potential impact of the lockdown on the duration of hospitalization and the final outcome. A total of 161 patients were included to our analysis with 1:1 male:female ratio and mean age 70.86 years. Most of the cases experienced gastrointestinal tract bleeding, biliary stone disease manifestations, or gastrointestinal malignancy complications, and 85.1% were discharged. Fewer patients were hospitalized during the lockdown period (40%), whereas the duration of hospitalization was significantly longer (7.69±4.55 vs. 5.76±4.36 days). Binary logistic regression analysis and sensitivity analysis demonstrated that the quarantine was associated with increased prevalence of negative outcomes (odds ratio 5.21, 95% confidence interval 1.66-16.34; P=0.005), especially in cases with gastrointestinal malignancy and acute pancreatitis (P=0.045 and P=0.041, respectively). The increase in the negative outcomes of common gastrointestinal diseases and the duration of hospitalization during the lockdown raise reasonable concerns regarding healthcare policies against further outbreaks.


Plasma kallikrein-kinin system in septicemia.
Plasma prekallikrein and functional kallikrein inhibition were studied in 18 surgical patients with complicating septicemia using chromogenic peptide substrate assays. Nine patients died and nine survived. In all 18 patients plasma prekallikrein values were reduced markedly when septicemia was diagnosed. During treatment gradually increasing values were found in the survivors, whereas values remained low in the fatal cases. Significantly reduced functional plasma kallikrein inhibition was associated with the development of fatal septic shock. The findings show that determination of these components of the plasma kallikrein-kinin system gives valuable information of prognostic value in patients with septicemia. Furthermore, the chromogenic peptide substrate assays used are fast and easy to perform and therefore suitable for intensive care medicine.


Clinical experience with daptomycin: bacteraemia and endocarditis.
Serious infections due to Staphylococcus aureus, especially those due to methicillin-resistant S. aureus, have become a major challenge. Vancomycin has long been the drug of choice for treatment of such infections, but failures due to its slow bactericidal activity coupled with increasing MICs have necessitated a search for new, more effective agents. Daptomycin has been studied by a number of investigators and has proved to be effective for bacteraemic infections due to staphylococci as well as vancomycin-resistant enterococci and other Gram-positive organisms. In addition, in a randomized controlled trial comparing daptomycin monotherapy with potentially synergistic therapy with either vancomycin or beta-lactam, both used in combination with an aminoglycoside, daptomycin achieved comparable outcomes but with significantly less nephrotoxicity.


Nuclear lamins: building blocks of nuclear structure and function.
The cell nucleus is surrounded by a complex membranous envelope which separates the nucleoplasm from the cytoplasm. Unlike the cytoplasm, the nucleoplasm is not subdivided into membrane-bound compartments, which allows for the efficient segregation of a wide range of complex metabolic activities. In the absence of such membrane compartmentalization, the nucleus is faced with the daunting task of efficiently segregating and interconnecting an enormous array of critically important functions. These include the assembly of the large multi-component complexes or 'factories' involved in DNA replication and transcription. These structures are dynamic as they are assembled and disassembled both spatially and temporally at different times, implying the existence of an infrastructure or nucleoskeleton responsible for establishing and maintaining a complex nuclear architecture. There is increasing evidence that the nuclear lamins are essential elements of this nuclear infrastructure, and that their proper assembly and organization are required for numerous essential nuclear functions. Our goal has been to determine the roles of the nuclear lamins in vital nuclear processes including DNA replication and transcription. The hypothesis directing our investigations is that the lamins form a 3D network that courses throughout the nucleoplasm providing an infrastructure for the assembly and distribution of numerous multicomponent complexes involved in a wide range of nuclear functions.


A narrative review of fluorescence imaging in robotic-assisted surgery.
In this review, we provide examples of applications of fluorescence imaging in urologic, gynecologic, general, and endocrine surgeries. While robotic-assisted surgery has helped increase the availability of minimally invasive procedures across surgical specialties, there remains an opportunity to reduce adverse events associated with open, laparoscopic, and robotic-assisted methods. In 2011, fluorescence imaging was introduced as an option to the da Vinci Surgical System, and has been standard equipment since 2014. Without interfering with surgical workflow, this fluorescence technology named Firefly® allows for acquisition and display of near-infrared fluorescent signals that are co-registered with white light endoscopic images. As a result, robotic surgeons of all specialties have been able to explore the clinical utility of fluorescence guided surgery. Literature searches were performed using the PubMed and MEDLINE databases using the keywords "robotic-assisted fluorescence surgery", "ICG robotic surgery", and "fluorescence guided surgery" covering the years 2011-2020. Real-time intraoperative fluorescence guidance has shown great potential in helping guide surgeons in both simple and complex surgical interventions. Indocyanine green is one of the most widely-used imaging agents in fluorescence guided surgery, and other targeted, near-infrared imaging agents are in various stages of development. Fluorescence is becoming a reliable tool that can help surgeons in their decision-making process in some specialties, while explorations continue in others.


Thyrotrophin receptor protein expression in normal and adenomatous human pituitary.
Thyrotrophin (TSH) synthesis and secretion is under the positive control of thyrotrophin releasing hormone and under the negative control of the thyroid hormones. However, it is hypothesised that TSH has a direct effect on the regulation of its own synthesis through an intrapituitary loop mediated by pituitary TSH receptors (TSH-R). The aim of this investigation was to study the expression of TSH-R in normal human pituitary at mRNA and protein levels, and to compare the pattern of protein expression between different pituitary adenomas. Using RT-PCR we were able to detect TSH-R mRNA in the normal pituitary, and immunohistochemical studies showed TSH-R protein expression in distinct areas of the anterior pituitary. Double immunostaining with antibodies against each of the intrapituitary hormones and S100 revealed that TSH-R protein is present in thyrotrophs and folliculostellate cells. Examination of 58 pituitary adenomas, including two clinically active and two clinically inactive thyrotroph adenomas, revealed TSH-R immunopositivity in only the two clinically inactive thyrotroph adenomas. This study shows, for the first time, the presence of TSH-R protein in the normal anterior pituitary and in a subset of thyrotroph adenomas. The expression of TSH-R in the thyrotroph and folliculostellate cell subpopulations provides preliminary evidence of a role for TSH in autocrine and paracrine regulatory pathways within the anterior pituitary gland.


Drug development of intranasally delivered peptides.
Intranasal drug delivery has attracted increasing attention as a noninvasive route of administration for therapeutic proteins and peptides. The delivery of therapeutic peptides through the nasal route provides an alternative to intravenous or subcutaneous injections. This review highlights the drug-development considerations unique to nasal therapeutics and discusses some of the factors and strategies that affect and can improve nasal absorption of peptides. The selectivity and good safety profile typical of peptide therapeutics, along with the dose limitation for intranasal administration, can provide challenges in drug development. Therefore, nasal peptide therapeutics often require special considerations in the nonclinical safety evaluations, such as determining drug exposure in the context of the maximum feasible dose in order to adequately prepare nasal products for clinical studies.


A second gene at the tomato Cf-4 locus confers resistance to cladosporium fulvum through recognition of a novel avirulence determinant
The tomato Cf-4 and Cf-9 genes confer resistance to the leaf mould pathogen Cladosporium fulvum and map at a complex locus on the short arm of chromosome 1. It was previously shown that the gene encoding Cf-4, which recognizes the Avr4 avirulence determinant, is one of five tandemly duplicated homologous genes (Hcr9-4s) at this locus. Cf-4 was identified by molecular analysis of rare Cf-4/Cf-9 disease-sensitive recombinants and by complementation analysis. The analysis did not exclude the possibility that an additional gene(s) located distal to Cf-4 may also confer resistance to C. fulvum. We demonstrate that a number of Dissociation-tagged Cf-4 mutants, identified on the basis of their insensitivity to Avr4, are still resistant to infection by C. fulvum race 5. Molecular analysis of 16 Cf-4 mutants, most of which have small chromosomal deletions in this region, suggested the additional resistance specificity is encoded by Hcr9-4E. Hcr9-4E recognizes a novel C. fulvum avirulence determinant that we have designated Avr4E.


Lycopene Improves the Metformin Effects on Glycemic Control and Decreases Biomarkers of Glycoxidative Stress in Diabetic Rats.
Oxidative stress and exacerbated generation of advanced glycation end products (AGEs) participate in the onset of diabetic complications. Lycopene is a potent antioxidant; evidence accounts for its ability to mitigate diabetic disturbances, including the deleterious events of advanced glycation. Therefore, this carotenoid has emerged as a candidate to be used in combination with antidiabetic drugs, such as metformin, attempting to counteract the glycoxidative stress. This study investigated the effects of the treatments with lycopene or metformin, alone or in combination, on glycoxidative stress biomarkers and antioxidant defenses in diabetic rats. Streptozotocin-induced diabetic rats were treated for 35 days with lycopene (45 mg/kg) or metformin (250 mg/kg), alone or as mixtures in yoghurt. Plasma levels of glucose, triglycerides, cholesterol, thiobarbituric acid reactive substances and protein carbonyl groups (biomarkers of oxidative damage), fluorescent AGEs (biomarkers of advanced glycation), and paraoxonase 1 activity (antioxidant enzyme) were assessed. Changes in the hepatic and renal levels of glycoxidative damage biomarkers and the activities of antioxidant enzymes were investigated. The combination of lycopene with metformin maintained the beneficial effects of the isolated treatments, improving the glucose tolerance and lipid profile, lessening biomarkers of oxidative damage, and increasing the paraoxonase 1 activity. Besides, the combined therapy caused further decreases in postprandial glycemia, plasma levels of cholesterol and AGEs, avoided lipid peroxidation (plasma, kidney), and increased antioxidant defenses, mainly the activity of superoxide dismutase (liver, kidney), indicating the maintenance of the lycopene effects. Lycopene combined with metformin may act synergistically in the control of postprandial glycemia, dyslipidemia and glycoxidative stress, as well as increased antioxidant defenses, arising as a promising therapeutic strategy to mitigate diabetic complications.