An ATP- and Ca2+-regulated Na+ channel in isolated intestinal epithelial cells.
When isolated intestinal epithelial cells are treated with 2 mM ATP, the unidirectional influx of Na+ to those cells increases from values near 50 to rates over 200 nmol . min-1 . mg protein-1. Calcium influx increases from 1 to 40 nmol . min-1 . mg protein-1. Within 2 min, the total cell Na+ increases two- to threefold, and total Ca+ increases about fivefold. The cells lose a major part of their capability for accumulating sugars during this interval. About 2 min after the time of ATP addition the normal permeability for Na+ and Ca2+ is restored, at which time the previously accumulated ions are rapidly extruded on a net basis until control levels are attained and the cells regain their usual sugar transport capability. The "repair" process requires Ca2+ in the incubation medium and is dependent on cellular uptake of Ca2+. Chlorpromazine (0.5 mM) blocks the Ca2+ entry route and the restoration of normal Na+ permeability. The Na+ entry route is selectively blocked by 4-acetamido-4'-isocyanostilbene-2,2'-disulfonic acid. The data show that ATP induces the influx of Na+ and Ca2+ by two different routes, which can be selectively inhibited. These ion flux routes may be involved in the events that allow intestinal tissue to convert from an absorptive state to a state in which net ion secretion occurs.


Injury-induced cold sensitization in Drosophila larvae involves behavioral shifts that require the TRP channel Brv1.
Nociceptive sensitization involves an increase in responsiveness of pain sensing neurons to sensory stimuli, typically through the lowering of their nociceptive threshold. Nociceptive sensitization is common following tissue damage, inflammation, and disease and serves to protect the affected area while it heals. Organisms can become sensitized to a range of noxious and innocuous stimuli, including thermal stimuli. The basic mechanisms underlying sensitization to warm or painfully hot stimuli have begun to be elucidated, however, sensitization to cold is not well understood. Here, we develop a Drosophila assay to study cold sensitization after UV-induced epidermal damage in larvae. Larvae respond to acute cold stimuli with a set of unique behaviors that include a contraction of the head and tail (CT) or a raising of the head and tail into a U-Shape (US). Under baseline, non-injured conditions larvae primarily produce a CT response to an acute cold (10°C) stimulus, however, we show that cold-evoked responses shift following tissue damage: CT responses decrease, US responses increase and some larvae exhibit a lateral body roll (BR) that is typically only observed in response to high temperature and noxious mechanical stimuli. At the cellular level, class III neurons are required for the decrease in CT, chordotonal neurons are required for the increase in US, and chordotonal and class IV neurons are required for the appearance of BR responses after UV. At the molecular level, we found that the transient receptor potential (TRP) channel brivido-1 (brv1) is required for these behavioral shifts. Our Drosophila model will allow us to precisely identify the genes and circuits involved in cold nociceptive sensitization.


Evaluation of dual-energy CT for differentiating intracerebral hemorrhage from iodinated contrast material staining.
To evaluate the efficacy of dual-energy computed tomography (CT) in the differentiation of intracerebral hemorrhage (ICH) from iodinated contrast material in patients who received contrast material via intraarterial or intravenous delivery. This retrospective study was approved by the local institutional review board, which waived the informed consent requirement for the analysis. Sixteen patients with acute stroke and two with head trauma who had undergone intraarterial or intravenous administration of iodinated contrast material were evaluated by using dual-energy CT to differentiate areas of hyperattenuation secondary to contrast material staining from those representing ICH. A dual-energy CT scanner was used for imaging at 80 and 140 kV, and a three-material decomposition algorithm was used to obtain virtual unenhanced images and iodine overlay images. The sensitivity, specificity, and accuracy of dual-energy CT in the prospective differentiation of intraparenchymal contrast material from hemorrhage were obtained. Follow-up images were used as the standard of reference. There were 28 intraparenchymal areas of hyperattenuation classified at dual-energy CT as iodinated contrast material staining (n = 20, 71%), hemorrhage (n = 5, 18%), or both (n = 3, 11%). Two of the three areas of hyperattenuation seen on both virtual unenhanced and iodine overlay images were related to mineralization. The sensitivity, specificity, and accuracy of dual-energy CT in the identification of hemorrhage were 100% (six of six areas), 91% (20 of 22 areas), and 93% (26 of 28 areas), respectively. Dual-energy CT can help differentiate ICH from iodinated contrast material staining with high sensitivity and specificity in patients who have recently received intraarterial or intravenous iodinated contrast material.


Interleukin-12 induces salivary gland dysfunction in transgenic mice, providing a new model of Sjögren's syndrome.
Interleukin-12 (IL-12) is a pleiotropic cytokine that is elevated in the affected organs of patients with Sjögren's syndrome (SS). We have previously reported that overexpression of IL-12 in CBA mice leads to mononuclear infiltration of salivary and lacrimal glands, as well as to expansion of bronchial lymphoid tissue and decreased mucociliary clearance. Because xerostomia is one of the most important clinical features in SS patients, our main objective in the current study was to evaluate salivary gland function in IL-12-transgenic mice. Our secondary objective was to further characterize this animal model and to determine if the changes observed in these mice are representative of those observed in patients with SS overall. Pilocarpine-stimulated salivary flow was used to address salivary gland function in a large group of IL-12-transgenic mice bred onto the autoimmune-prone SJL background. Furthermore, salivary glands were removed to assess the formation of infiltrates in the glands and gland morphology. Serum was also collected from these animals to investigate the formation of autoantibodies. Pilocarpine-stimulated salivary flow was significantly lower in IL-12-transgenic mice than in wild-type controls. Salivary glands from transgenic mice exhibited an increase in both the number and the size of lymphocytic foci, versus glands from age-matched controls. Furthermore, the acini in transgenic mice were fewer in number and larger in size compared with acini in controls. An age-dependent increase in anti-SSB/La antibodies was observed in IL-12-transgenic mice and was accompanied by an increase in antinuclear antibodies. Our findings indicate that a number of conditions associated with SS are exhibited by IL-12-transgenic SJL mice and that this model might be useful in researching multiple aspects of the disease.


Morganella morganii: epidemiology of bacteremic disease.
A retrospective review of microbiology records revealed 19 documented episodes of M. morganii bacteremia in 18 patients at a Veterans Administration hospital during a 5.5 year period. Thirteen of 19 bacteremias were related to nosocomial infections; 11 of the 13 nosocomial bacteremias occurred in surgical patients. Nine of the 13 patients with nosocomial bacteremia had received recent therapy with a beta-lactam antibiotic. The most common source of bacteremia was a postoperative wound infection (seven episodes). Only one episode was related to a urinary tract infection. Retrospective analysis showed that clusters of cases of M. morganii bacteremia had occurred almost yearly. This finding prompted a six-month period of prospective monitoring of all cultures for M. morganii to identify human reservoirs in our institution. Sixty percent of all cultures growing M. morganii came from urine cultures, 18% came from wound cultures, and the remaining 22% came from a variety of body fluids or tube drainage. Thirty-one percent of patients harboring M. morganii were on the Surgical Service. M. morganii bacteremia most commonly occurs in postoperative patients who receive beta-lactam antibiotics. From the data in this study, M. morganii is an infrequent cause of bacteremia, and its presence in blood cultures may be an indicator of an environment conducive for an outbreak of nosocomial infection.


The Glanzmann's Thrombasthenia in Tunisia: A Cohort Study.
The Glanzmann's thrombasthenia (GT) is a rare autosomal-recessive bleeding disorder with uncommon neonatal revelation. It is due to abnormalities of quantitative and/or qualitative αIIbβ3 integrin. This cell adhesion receptor is essential for platelet aggregation and allows the formation of a hemostatic plug if the vessel is damaged by injury. The clinical picture of GT is variable, with mucocutaneous bleeding due to non-functional platelets. Management requires a good expertise in bleeding disorders. We describe the clinical and the epidemiological data of GT in Aziza Othmana Hospital Hemophilia Center. This was a retrospective study of all patients with GT monitored and treated in our hemophilia center during the period of 2011 - 2015. Twenty-seven patients among the 35 patients included in our hemophilia center registry were studied. The most common sign encountered is the gingival bleeding. In our women cohort, one completed her pregnancy. The consanguinity is present with a frequency of 62%. Treatments used depending on the case are tranexamic acid, platelet transfusion, packed red blood cells and rFVIIa, respectively. GT is relatively frequent in Tunisia and especially in the North of the country which can be explained by the high consanguinity in our population.


Tai Chi Chuan exercise related change in brain function as assessed by functional near-infrared spectroscopy.
Early studies have shown that Tai Chi Chuan (TCC) contributes to the rehabilitation of cognitive disorders and increases blood oxygen concentration levels in the parietal and occipital brain areas; however, the mechanism of TCC training on brain function remains poorly understood. This study hypothesize that TCC has altered brain function and aims to explore the effects of TCC on functional connection and effective connection of the prefrontal cortex (PFC), motor cortex (MC), and occipital cortex (OC). The participants were 23 experienced Chen-style TCC practitioners (TCC group), and 32 demographically matched TCC-naive healthy controls (control group). Functional and effective connections were calculated using wavelet-based coherence analysis and dynamic Bayesian inference method, respectively. Results showed that beyond the intensity of activity in a particular cortical region induced by TCC, significant differences in brain activity and dynamic configuration of connectivity were observed between the TCC and control groups during resting and movement states. These findings suggested that TCC training improved the connection of PFC, MC and OC in myogenic activity, sympathetic nervous system, and endothelial cell metabolic activities; enhanced brain functional connections and relayed the ability of TCC to improve cognition and the anti-memory decline potential.


Switching rates in health insurance markets decrease with age: empirical evidence and policy implications from the Netherlands.
All consumer groups with specific preferences must feel free to easily switch insurer in order to discipline insurers to be responsive to consumers' heterogeneous preferences. This paper provides insight into the switching behaviour of low-risks (i.e. young or healthy consumers) and high-risks (i.e. elderly or unhealthy consumers) in the Netherlands in the period 2009-2012. We analysed: (1) administrative data with objective health status information (i.e. medically diagnosed diseases and pharmaceutical use) and information on health care expenses of nearly the entire Dutch population (n=15.3 million individuals) and (2) three-year sample data (n=1152 individuals). Our findings indicate that switching rates strongly decrease with age. For example, in 2009, consumers aged 25-44 switched 10 times more than consumers aged 75 or older. Another finding is that switching rates decrease as the predicted health care expenses increase. Although healthy consumers switch twice as much as unhealthy consumers, this difference becomes much smaller after adjusting for age. We conclude that our findings can be explained by higher perceived switching costs by elderly consumers than by young consumers. Consequently, insurers have low incentives to act as quality-conscious purchasers of care for the elderly consumers. Therefore, strategies should be developed to increase the choice of insurer of elderly consumers.


Zirconium-Catalyzed Hydroaminoalkylation of Alkynes for the Synthesis of Allylic Amines.
A zirconium-catalyzed hydroaminoalkylation of alkynes to access α,β,γ-substituted allylic amines in an atom-economic fashion is reported. The reaction is compatible with N-(trimethylsilyl)benzylamine and a variety of N-benzylaniline substrates, with the latter giving the allylic amine as the sole organic product. Various internal alkynes with electron-withdrawing and electron-donating substituents were tolerated. Model intermediates of the reaction were synthesized and structurally characterized. Stoichiometric studies on key intermediates revealed that the open coordination sphere at zirconium, imparted by the tethered bis(ureate) ligand, is crucial for the coordination of neutral donors. These complexes may serve as models for the inner-sphere protonolysis reactions required for catalytic turnover.


Blue macules of localized argyria caused by implanted acupuncture needles. Electron microscopy and roentgenographic microanalysis of deposited metal.
Acupuncture needles implanted in the skin for more than ten years caused peculiar bluish macules, each of which clinically resembled a blue nevus in the extremities of a 63-year-old Japanese woman. Histologically, the involved skin showed deposition of fine brownish granules in the basement membrane of the eccrine sweat glands, on the inner surface of the blood vessel walls, and along elastic fibers of the superficial dermis in addition to sparse deposits noted throughout the dermis. Electron microscopy revealed deposits of electron-dense particles on the basal lamina of the secretory coils of the eccrine sweat glands, below the basal lamina of the dermoepidermal junction, and on elastic fibers. Roentgenographic microanalysis of the involved skin demonstrated that most of the granules consisted of silver and chloride; silver was a major component in the removed needles.


Pregnancy specific beta 1 glycoprotein: a comparison of analysis by radioimmunoassay and enzyme linked immunosorbent assay observations on patients with malignant teratoma of testis.
A comparison of serum pregnancy specific beta 1 glycoprotein analysis by enzyme linked immunosorbent assay and radioimmunoassay has been made. Excellent correlation was obtained between the two methods but radioimmunoassay was found to be more precise and to have a lower detection limit than enzyme linked immunosorbent assay. Using the more sensitive radioimmunoassay system, 2 out of 10 control male sera were found to contain detectable levels of pregnancy specific beta 1 glycoprotein. Sequential results are presented on patients with diagnosed malignant teratoma of testis who were undergoing chemotherapy.


Echocardiographic method for the estimation of pulmonary artery pressure in chronic lung disease.
The prognostic implications of pulmonary hypertension in chronic lung disease, and the difficulty in establishing its severity without cardiac catheterisation, indicate the need for a reliable non-invasive method of assessing the pulmonary artery pressure. It is likely that the time taken by the right ventricle to generate a sufficiently high pressure to open the pulmonary valve will increase progressively as the pulmonary arterial diastolic pressure rises. Therefore, the time interval between closure of the tricuspid valve and opening of the pulmonary valve has been obtained by high-speed echocardiographic recordings of the tricuspid and pulmonary valves in a group of 17 patients with chronic lung disease. Each patient underwent right heart catheterisation immediately after the echocardiographic examination so that the pulmonary arterial diastolic pressure could be obtained directly. A range for the group from 15 mmHg (2.0 kPa) to 45 mmHg (6.0 kPa) was observed. The linear correlation between the measured diastolic pressure and time interval from the tricuspid valve closure to pulmonary valve opening was highly significant (r = 0.94, p = less than 0.001) and the scatter was relatively small. It is therefore suggested that this time interval, obtained non-invasively by echocardiography, can be used as an index of the severity of pulmonary hypertension associated with chronic lung disease.


Subaltern biology? Local biologies, Indian odysseys, and the pursuit of human embryonic stem cell therapies.
As an emerging life form, stem cells are viewed as global biological entities. This extends Margaret Lock's categorization of local biological or local biologies as emerging from debates about menopause, brain death, and organ transplant. I seek to reconceptualize the local, shape-shifting nature of the biological form and the ontological mutations as inherently subaltern. I draw on a multisited ethnography and a spectrum of global publics from more than 22 countries in search of contentious human embryonic stem cell therapies (hESC) in India. In so doing, I show that the subaltern local biological experience seldom finds utterance and acknowledgement in the face of hegemonic, universalized, epistemic discourse on human body and biological form.


Dynamic properties of force fields.
Molecular-dynamics simulations are increasingly used to study dynamic properties of biological systems. With this development, the ability of force fields to successfully predict relaxation timescales and the associated conformational exchange processes moves into focus. We assess to what extent the dynamic properties of model peptides (Ac-A-NHMe, Ac-V-NHMe, AVAVA, A10) differ when simulated with different force fields (AMBER ff99SB-ILDN, AMBER ff03, OPLS-AA/L, CHARMM27, and GROMOS43a1). The dynamic properties are extracted using Markov state models. For single-residue models (Ac-A-NHMe, Ac-V-NHMe), the slow conformational exchange processes are similar in all force fields, but the associated relaxation timescales differ by up to an order of magnitude. For the peptide systems, not only the relaxation timescales, but also the conformational exchange processes differ considerably across force fields. This finding calls the significance of dynamic interpretations of molecular-dynamics simulations into question.


The decision to operate in rheumatoid arthritis.
The physician caring for the patient with rheumatoid arthritis should not look upon surgery as a last resort to be turned to when all else has failed and the situation is desperate. Surgical intervention may be important at any stage in the disease. Wrist fusion to obliterate pain and to restore a strong grip is advantageous whenever pain and instability compromise the function of that extremity. The knee with good painless flexion and extension cannot help but insure the patient's ability to get up and down from a chair. There are many goals that may be achieved through surgical intervention, and each of these should be considered and reviewed every time the physician sees the patient. Consider the benefits of prevention of disease progression, the stopping of bone resorption, the relief of pain, the restoration of motion, strength, and stability, and the preservation or restoration of the patient's confidence and determination to remain an independent member of society.


Inhibition of effects of flow on potassium permeability in single perfused frog mesenteric capillaries.
1. We have investigated the effects of various potential inhibitors on flow-dependent K+ permeability (PK) of single perfused mesenteric microvessels in pithed frogs. 2. Neither superfusion with a nitric oxide synthase inhibitor, NG-monomethyl-L-arginine (10 or 100 micromol l-1), nor the addition of indomethacin (30 micromol l-1) to both perfusate and superfusate reduced the positive correlation between PK and flow velocity (U). 3. In the presence of agents known to raise intracellular levels of adenosine 3',5'-cyclic monophosphate (noradrenaline, 8-bromo-cAMP and a combination of forskolin and rolipram) the slope of the relation between PK and U was no longer significant, so that PK was no longer flow dependent. 4. These results confirm that the flow dependence of PK is a biological process and not an artefact of measurement and suggest a role for intracellular cAMP rather than nitric oxide or prostacyclin in the flow-dependent modulation of PK in frog mesenteric microvessels.


A closed conformation of Bacillus subtilis oxalate decarboxylase OxdC provides evidence for the true identity of the active site.
Oxalate decarboxylase (EC 188.8.131.52) catalyzes the conversion of oxalate to formate and carbon dioxide and utilizes dioxygen as a cofactor. By contrast, the evolutionarily related oxalate oxidase (EC 18.104.22.168) converts oxalate and dioxygen to carbon dioxide and hydrogen peroxide. Divergent free radical catalytic mechanisms have been proposed for these enzymes that involve the requirement of an active site proton donor in the decarboxylase but not the oxidase reaction. The oxidase possesses only one domain and manganese binding site per subunit, while the decarboxylase has two domains and two manganese sites per subunit. A structure of the decarboxylase together with a limited mutagenesis study has recently been interpreted as evidence that the C-terminal domain manganese binding site (site 2) is the catalytic site and that Glu-333 is the crucial proton donor (Anand, R., Dorrestein, P. C., Kinsland, C., Begley, T. P., and Ealick, S. E. (2002) Biochemistry 41, 7659-7669). The N-terminal binding site (site 1) of this structure is solvent-exposed (open) and lacks a suitable proton donor for the decarboxylase reaction. We report a new structure of the decarboxylase that shows a loop containing a 3(10) helix near site 1 in an alternative conformation. This loop adopts a "closed" conformation forming a lid covering the entrance to site 1. This conformational change brings Glu-162 close to the manganese ion, making it a new candidate for the crucial proton donor. Site-directed mutagenesis of equivalent residues in each domain provides evidence that Glu-162 performs this vital role and that the N-terminal domain is either the sole or the dominant catalytically active domain.


TP53 and EGFR Mutational Status in Thymoma: A Genetic Sequencing Study.
Thymoma is a rare malignant tumor that usually with an indolent presentation, which was falsely assumed to be benign previously. The tumor suppressor P53 (TP53) and EGFR gene mutate most frequently in human cancers. We tried to investigate the presence of TP53 and EGFR mutations among thymoma patients referred to an Indonesian referral respiratory hospital and to discuss its potential role in thymoma management and prognosis. Surgically resected tumor tissues were collected from thymoma patients and then underwent genomic analysis. PCR was performed on the extracted Paraffinized DNA to amplify exon 6 of TP53 and exons 18, 19, and 21 of EGFR. The evaluation of mutational status was done using direct sequencing and sequence analysis of purified PCR products. Among 27 collected samples, TP53 exon 6 mutation, namely missense mutation and nonsense mutation, was observed in two samples (7.4%). EGFR exon 18 mutation, namely E709K and nonsense mutation, was found in 2 samples (7.4%). An intronic mutation in EGFR exon 19 (3.7%) and exon 21 (3.7%) was observed in one sample. TP53 and EGFR mutations were not most frequent, so it seems that these genes are not involved in the pathogenesis of thymoma in Indonesian patients. Nevertheless, we found two samples with a significant mutation in p53 and EGFR genes, suggesting further research on thymoma prognostification and targeted therapy.


Zoledronate stimulates gamma delta T cells in prostate cancer patients.
Androgen deprivation therapy is the mainstay of treatment for prostate cancer. Given its frequent failure, new therapy that reduces prostate cancer progression would be a breakthrough in treating this disease. Bisphosphonates are well-established agents for treating skeletal-related events (SREs) in prostate cancer patients with bone metastases. Exposure to bisphosphonates may not only reduce the incidence of SREs, but also have anticancer effects by modulating a patient's immunity. The purpose of this study was to examine the effect of zoledronate (ZOL) on gamma delta T cells, serum prostate-specific antigen (PSA) levels, and velocities. The effect of ZOL, with and without IL-2, on gamma delta T cell activation was examined in vitro. Furthermore, the activated state and the number of gamma delta T cells and changes in serum PSA levels were examined for patients who received ZOL infusion for the prevention of SREs. We found that ZOL activated gamma delta T cells, and the number of gamma delta T cell was increased when IL-2 was administered with ZOL in vitro. Comparisons before and after the first ZOL infusion revealed that gamma delta T cells in peripheral blood were activated by ZOL. Moreover, after the first ZOL treatment, reduction in serum PSA was observed in 3 of 11 patients, and reduction in PSA velocity was observed in 5 of 10 patients. Our findings indicate that ZOL stimulates gamma delta T cells in vivo and in vitro. This study provides further insight into the ability of gamma delta T cells to induce an antitumor immune response.


[The characteristics of the effect of "reward zones" on the electrical activity of the parafascicular nuclear complex in the developing rabbit].
In 35 rabbits of 41-60-day age, the effect of electrical stimulation of the "rewarding areas" identified preliminarily by means of self-stimulation, on EPs of the thalamus parafascicular complex in response to peripheral nociceptive stimulation, was studied. Specifics of the effects was revealed for this particular age. In some parameters, the antinociceptive effects of the "rewarding areas" on the EPs in the parafascicular complex were less obvious in between the exaltation and adult periods of development.


Incremental Prognosis by Left Atrial Functional Assessment: The Left Atrial Coupling Index in Patients With Floppy Mitral Valves.
Background Emerging data suggest important prognostic value to left atrial (LA) characteristics, but the independent impact of LA function on outcome remains unsubstantiated. Thus, we aimed to define the incremental prognostic value of LA coupling index (LACI), coupling volumetric and mechanical LA characteristics and calculated as the ratio of left atrial volume index to tissue Doppler imaging a', in a large cohort of patients with isolated floppy mitral valve. Methods and Results All consecutive 4792 patients (61±16 years, 48% women) with isolated floppy mitral valve in sinus rhythm diagnosed at Mayo Clinic from 2003 to 2011, comprehensively characterized and with prospectively measured left atrial volume index and tissue Doppler imaging a' in routine practice, were enrolled, and their long-term survival analyzed. Overall, LACI was 5.8±3.7 and was <5 in 2422 versus ≥5 in 2370 patients. LACI was independently higher with older age, more mitral regurgitation (no 3.8±2.3, mild 5.1±3.0, moderate 6.5±3.8, and severe 7.8±4.3), and with diastolic (higher E/e') and systolic (higher end-systolic dimension) left ventricular dysfunction (all P≤0.0001). At diagnosis, higher LACI was associated with more severe presentation (more dyspnea, more severe functional tricuspid regurgitation, and elevated pulmonary artery pressure, all P≤0.0001) independently of age, sex, comorbidity index, ventricular function, and mitral regurgitation severity. During 7.0±3.0 years follow-up, 1146 patients underwent mitral valve surgery (94% repair, 6% replacement), and 880 died, 780 under medical management. In spline curve analysis, LACI ≥5 was identified as the threshold for excess mortality, with much reduced 10-year survival under medical management (60±2% versus 85±1% for LACI <5, P<0.0001), even after comprehensive adjustment (adjusted hazard ratio, 1.30 [95% CI, 1.10-1.53] for LACI ≥5; P=0.002). Association of LACI ≥5 with higher mortality persisted, stratifying by mitral regurgitation severity of LA enlargement grade (all P<0.001) and after propensity-score matching (P=0.02). Multiple statistical methods confirmed the significant incremental predictive power of LACI over left atrial volume index (all P<0.0001). Conclusions LA functional assessment by LACI in routine practice is achievable in a large number of patients with floppy mitral valve using conventional Doppler echocardiographic measurements. Higher LACI is associated with worse clinical presentation, but irrespective of baseline characteristics, LACI is strongly, independently, and incrementally determinant of outcome, demonstrating the crucial importance of LA functional response to mitral valve disease.


Critical issues facing animal scientists.
An important series of issues confronts the animal research scientist involved in meeting world needs for food products from animal agriculture. The major issues stem from societal pressures for improved products, for prevention of chemical pollution of the biosphere, and for wise use of animal resources in toxicological testing and research. The following issues are among those treated in the present discussion: (1) There is a need to obtain reliable dosage vs response information for chronic effects produced in the human by food additive chemicals. (2) There is a need for toxicological research on combined hazard from multiple contaminants in animal food products. (3) There is a requirement for development of methodology for proper usage and (or) disposal of chemically contaminated animal food products. (4) The public must be educated with respect to the absolute necessity for use of intact animals in biological research, to supplement information from in vitro systems. (5) Animal scientists must characterize fully each animal species and strain employed in research and testing, to guarantee reproducibility of results. (6) Animal scientists must evolve a universal set of principles for translation of animal toxicity data into prediction of human hazard. (7) Animal scientists should develop universally uniform methodology for toxicological testing of foods and food chemical residues. (8) Animal scientists should rebut national regulatory philosophies that derive from unsound science. (9) The animal scientist should assist regulatory authorities in developing sound, practical "Good Laboratory Practice" guidelines.


Renewable and non-renewable energy consumption, energy technology investment, green technological innovation, and environmental sustainability in the United States: Testing the EKC and LCC hypotheses with novel Fourier estimation.
The energy sector represents one of the key contributors to environmental degradation. In this context, actions taken within the energy sector are paramount in the global effort to combat climate change. This study aims to investigate the impacts of renewable and non-renewable energy consumption, energy technology investment, and green technological innovation on environmental sustainability in the context of EKC and LCC hypotheses in the USA from 1980 to 2015. While many studies in the literature focus on the EKC hypothesis, this study offers a comparative analysis of the EKC and LCC hypotheses with relevant variables. For this aim, the study uses the novel Fourier estimation methods. According to the results, the EKC and the LCC hypotheses are valid in the USA. Moreover, non-renewable energy consumption increases the ecological footprint. On the other hand, non-renewable energy consumption decreases the load capacity factor while renewable energy consumption increases it. Finally, energy technology investment and green technological innovation have an insignificant impact on the ecological footprint and the load capacity factor in both models. All results except energy technology investment and green technological innovation are consistent with our expectations. The USA has the potential to realize green growth. Policymakers should support the green technological innovation process and increase green investments.


Regeneration in MRL mice: further genetic loci controlling the ear hole closure trait using MRL and M.m. Castaneus mice.
The MRL mouse has been shown to display an epimorphic regenerative response after ear hole punching leading to complete closure within 30 days and cartilage regrowth. The regenerative capacity of the MRL has also been seen after a severe cryoinjury to the heart leads to complete healing without scarring and functional myocardium. The wound healing ear hole closure response that occurs in MRL mice has been shown to be genetically controlled. We have previously identified 11 quantitative trait loci (QTL) that govern healing in an intercross of (MRL x C57BL/6 J) mice. However, it is desirable to use another poorly healing mouse strain to elucidate the full range of genetic factors that affect this important process. In the current study, we have used an inbred subspecies of the mouse, M. castaneus, and have confirmed a number of loci identified previously. In addition, we report three new healing QTL. Furthermore, in this strain combination, we note a strong sexual dimorphism also observed in the MRL x C57BL/6 cross, both in the healing trait and in the QTL that control it.


Anthrax in Wabessa village in the Dessie Zuria district of Ethiopia.
In 2002 an investigation of sudden death in a goat in Wabessa village in the Dessie Zuria district of Ethiopia was undertaken using fresh blood brought to the Kombolcha Regional Veterinary Laboratory. The sample was examined using standard bacteriological techniques and animal pathogenicity tests were also performed. The laboratory investigation revealed Bacillus anthracis as the cause of sudden death. Information gathered from stockowners in the same village revealed other similar recent cases and deaths, both in animals and humans, with farmers clearly describing the clinical signs and necropsy findings of anthrax. The disease occurs annually in this area in May and June, and in the 2002 outbreak mortality rates of 7.7%, 32.7% and 47.1% were observed in cattle, goats and donkeys, respectively. This study indicates that the community of this particular village neither knows of, nor practises, any of the conventional methods for anthrax control. The cutaneous form of the disease in humans and the environmental contamination associated with the practise of opening cadavers are briefly described and the findings are discussed with reference to the epidemiology of anthrax in both Ethiopia and elsewhere. Control strategies are also recommended.


Recruiting and retaining a diverse workforce in nursing: from evidence to best practices to policy.
Health policy makers, providers, clinicians, and social scientists are among those who have identified racial and ethnic diversification of the health care workforce as one strategy for solving the seemingly intractable problem of health disparities in the U.S. population. But evidence supporting the impact of such diversification on narrowing health disparities is lacking, thus making it unclear if the push for workforce diversification is empirically or politically driven. Moreover, data are largely derived from the study of physicians, making it difficult to generalize findings to nursing and other health professions. This article reviews the evidence that supports the impact of a diverse workforce on patient outcomes and delivery services. Assuming a positive social value in the absence of the data, the authors review the approaches that have been successful in diversifying the nursing workforce. The authors conclude with recommendations for research and policies, including best practices, for enhancing recruitment and retention of a diverse nursing workforce.


First step in the development of an ultrasound joint inflammation score for rheumatoid arthritis using a data-driven approach.
To develop and validate candidate sets of joints and tendons for assessment of ultrasound (US) joint inflammation in rheumatoid arthritis (RA). Patients were included in one of two cohorts from 2010 to June 2013: disease-modifying antirheumatic drug naïve early RA or established RA starting/switching biologics. An extensive US examination was performed by experienced sonographers using a validated grey-scale (GSUS) and power Doppler (PDUS) semiquantitative scoring system with scores 0-3 for both GSUS and PDUS in 36 joints and four tendons. We performed factor analysis in the early RA US data and selected candidate joint/tendon sets based on these results. The proportion of information in the total US scores retained in these candidate sets was assessed by R(2) from linear regression analysis. Finally, the candidate sets and previously proposed joint scores were tested in the established RA cohort, and we also evaluated the sensitivity to change with standardised response means. 227 patients with early RA and 212 patients with established RA were included. We identified two candidate sets of joints/tendons: candidate set A consisted of seven joints/two tendons (meatacarpophalangeal 1 (MCP1), MCP2, proximal interphalangeal 3, radiocarpal, elbow, metatarsophalangeal 1 (MTP1), MTP2, tibialis posterior tendon, extensor carpi ulnaris tendon) and set B of nine joints/two tendons (MCP5 and MTP5 added to set A). Unilateral reduced scores retained 78%-85% of the information in total score, while bilateral reduced scores retained 89%-93%, and both sets performed better than previously proposed reduced joint scores, and similar or slightly better regarding sensitivity to change. The reduced GSUS and PDUS scores retained most of the information from the total score and performed well in a validation cohort of established RA. NCT01205854, ACTRN12610000284066.


Cardiopulmonary exercise performance in prematurely born children.
Prematurely born children have reduced peak VO2 compared with their peers, inferentially attributed to ventilatory limitation. The primary purpose of this study was to compare exercise ventilation and cardiac output in a sample of childhood survivors of lung disease of prematurity with those of a control group to elucidate reasons for lower peak VO2. A secondary aim was to describe and compare the ventilatory response to incremental exercise. Thirty-two children, aged 8-9 y, were recalled for lung function and progressive exercise tests. Fifteen of them also performed submaximal exercise with measurement of cardiac output (indirect [CO2] Fick) and physiologic dead space. Results were compared with those of term-born, age- and sex-matched, control children. Pulmonary function tests showed mild airflow limitation. Peak VO2 was lower in prematurely born children compared with control children, and was correlated with lean body mass. Their heart rate-VO2 relationship and stroke volume were similar to that of term-born control children. Children with a history of bronchopulmonary dysplasia and hyaline membrane disease as infants exhibited greater exercise hyperpnea than did healthy control children, because of higher breathing frequency, and maintained lower end-tidal PCO2 during submaximal exercise. Physiologic dead space normalized for body weight was similar in preterm and term-born children. Lower peak VO2 in this population is not caused by cardiopulmonary factors, but is best predicted by lean body mass. Ventilation did not limit exercise performance, although it appears that breathing during exercise is regulated differently in prematurely born children than in term-born children.


Neuropsychiatric aspects of trace elements.
Our knowledge of trace element metabolism has increased dramatically in the last decade. It now seems likely that many trace elements are of significance either as causative or therapeutic factors in a wide range of illnesses. Several of the most topical elements are discussed here in relation to neurological and psychiatric disorders.


[An electrophysiological analysis of pathways from lateral septum to hippocampal CA1 area in rat].
Upon stimulating the lateral septal region, a population spike could be recorded in the stratum pyramidale of hippocampal CA1 together with a recordable EPSP in the apical dendrites (stratum radiatum). After destruction of hippocampal CA3, CA4 pyramidal cells by intraventricular microinjection of kainic acid, electrical stimulation of the lateral septum could no longer evoke EPSP in the apical dendrites. This experiment suggests that the neural connection of the lateral septo-CA1 apical dendrites is mediated by the collaterals of the ipsilateral CA3 pyramidal cells. But after the destruction of CA3, CA4, electric stimulation of lateral septum could still evoke EPSP in the basal dendrites (stratum oriends). The neural connection mediating the response of the lateral septo-CA1 basal dendrites was not reported before, but the neural pathway remains to be investigated.


Cell-penetrating metal complex optical probes: targeted and responsive systems based on lanthanide luminescence.
To understand better the structure and function of biological systems, cell biologists and biochemists would like to have methods that minimally perturb living systems. The development of emissive optical probes is essential for improving our observation of intracellular signaling and recognition processes. Following excitation of the probe, photons emitted from the probe may be observed by spectroscopy or microscopy and encode information about their environments in their energy, lifetime, and polarization. Such optical probes may be based on organic fluorophores, quantum dots, recombinant proteins, or emissive metal complexes. In this Account, we trace the emergence of lanthanide coordination complexes as emissive optical probes. These probes benefit from sharp emission bands and long lifetimes. We can design these complexes to report on the concentration of key biochemical variables by modulation of spectral form, lifetime, or circular polarization. These properties allow us to apply ratiometric methods of analysis in spectroscopy or microscopy to report on local pH, pM (M = Ca, Zn), or the concentration of certain anionic metabolites, such as citrate, lactate, bicarbonate, or urate. For optical microscopy studies in living cells, these probes must be cell-permeable and, ideally, should localize in a given cell organelle. We undertook systematic studies of more than 60 emissive complexes, examining the time dependence of cellular uptake and compartmentalization, cellular toxicity, protein affinity, and quenching sensitivity. These results and their relationship to probe structure have allowed us to identify certain structure-activity relationships. The nature and linkage mode of the integral sensitizing group-introduced to harvest incident light efficiently-is of primary importance in determining protein affinity and cellular uptake and trafficking. In many cases, uptake may occur via macropinocytosis. We have defined three main classes of behavior: complexes exhibit predominant localization profiles in protein-rich regions (nucleoli/ribosomes), in cellular mitochondria, or in endosomes/lysosomes. Therefore, these systems offer considerable promise as intracellular optical probes, amenable to single- or two-photon excitation, that may report on the local ionic composition of living cells subjected to differing environmental stresses.


Pharmacologic management of Duchenne muscular dystrophy: target identification and preclinical trials.
Duchenne muscular dystrophy (DMD) is an X-linked human disorder in which absence of the protein dystrophin causes degeneration of skeletal and cardiac muscle. For the sake of treatment development, over and above definitive genetic and cell-based therapies, there is considerable interest in drugs that target downstream disease mechanisms. Drug candidates have typically been chosen based on the nature of pathologic lesions and presumed underlying mechanisms and then tested in animal models. Mammalian dystrophinopathies have been characterized in mice (mdx mouse) and dogs (golden retriever muscular dystrophy [GRMD]). Despite promising results in the mdx mouse, some therapies have not shown efficacy in DMD. Although the GRMD model offers a higher hurdle for translation, dogs have primarily been used to test genetic and cellular therapies where there is greater risk. Failed translation of animal studies to DMD raises questions about the propriety of methods and models used to identify drug targets and test efficacy of pharmacologic intervention. The mdx mouse and GRMD dog are genetically homologous to DMD but not necessarily analogous. Subcellular species differences are undoubtedly magnified at the whole-body level in clinical trials. This problem is compounded by disparate cultures in clinical trials and preclinical studies, pointing to a need for greater rigor and transparency in animal experiments. Molecular assays such as mRNA arrays and genome-wide association studies allow identification of genetic drug targets more closely tied to disease pathogenesis. Genes in which polymorphisms have been directly linked to DMD disease progression, as with osteopontin, are particularly attractive targets.


Carbohydrates in individual poplar fine roots: effects of root age and defoliation.
Late-summer starch accumulation in fine roots of poplars (Populus x canadensis Moench.) defoliated by gypsy moth (Lymantria dispar L.) lagged behind that in fine roots of undefoliated trees. If starch concentration declines with age, defoliation-induced changes in root system age structure could be partly responsible for this difference. To test this hypothesis, we measured fine-root starch and soluble sugar concentrations in roots of known age from trees in defoliated and undefoliated plots. There was a significant interaction between the effects of defoliation and root type (white, brown, or dead) on fine root soluble sugar concentration because of the high concentration of soluble sugars in white roots from trees in undefoliated plots. Both root starch and soluble sugar concentrations were variable among individuals of each age class. The population frequency distributions for starch and soluble sugar concentrations were both right-skewed, and fit by exponential functions. These data are most consistent with direct defoliation effects on a labile and dynamic pool of carbohydrates in poplar fine roots, rather than indirect defoliation effects on root system age structure.


The inhibitory effect of genistein on hepatic steatosis is linked to visceral adipocyte metabolism in mice with diet-induced non-alcoholic fatty liver disease.
Non-alcoholic fatty liver disease (NAFLD) has been deeply associated with visceral adiposity, adipose tissue inflammation and a variety of adipocytokines. We reported previously that genistein inhibited NAFLD by enhancing fatty acid catabolism. However, this molecular approach focused on hepatic metabolism. Thus, we have attempted to determine whether this anti-steatotic effect of genistein is linked to visceral adipocyte metabolism. C57BL/6J mice were fed on normal-fat (NF) diet, high-fat (HF) diet and HF diet supplemented with genistein (1, 2 and 4 g/kg diet) for 12 weeks. Mice fed on the HF diet gained body weight, exhibited increased visceral fat mass and elevated levels of serum and liver lipids, and developed NAFLD, unlike what was observed in mice fed on the NF diet. However, genistein supplementation (2 and 4 g/kg diet) normalised these alternations. In the linear regression analysis, visceral fat (R 0·77) and TNFα (R 0·62) were strongly correlated with NAFLD among other NAFLD-related parameters. Genistein supplementation suppressed the hypertrophy of adipocytes via the up-regulation of genes involved in fatty acid β-oxidation, including PPARα, 5'-AMP-activated protein kinase and very long-chain acyl CoA dehydrogenase, as well as through the down-regulation of genes associated with adipogenesis or lipogenesis, including liver X receptor-α, sterol-regulatory element-binding protein-1c, PPARγ, retinoid X receptor-α and acetyl CoA carboxylase 2. Moreover, genistein supplementation augmented an anti-steatohepatitic adiponectin TNF and reduced a steatohepatitic TNFα. Collectively, these findings show that genistein may prevent NAFLD via the regulation of visceral adipocyte metabolism and adipocytokines.


Long-term functional outcomes in the elderly after burn injury.
Although the elderly represent a substantial proportion of the population, limited information exists on postdischarge long-term outcomes of elderly burn survivors. The purpose of this study was to assess elderly burn patient outcomes 2 to 10 years after discharge. This study was a prospective cross-sectional survey assessment of quality of life and retrospective trauma registry for the American College of Surgeons review of patients ≥ 60 years of age discharged alive after acute burn from 1997 to 2007. In-hospital treatment and burn demographic information were obtained from database and chart review. Surviving patients or their families were contacted, and the Short-Form-12 and Functional Independence Measure (FIM) administered. Of the 344 patients discharged, 232 participated. Mean age was 72.3 (60-85.8) years, TBSA burn was 7.8% (1-79), and length of stay was 11.2 ± 0.9 days (1-51). Most patients were discharged home (71%) or to a skilled nursing facility (SNF; 20%). Mean interval between discharge and survey administration was 46.1 months. In all, 24% of patients sent home died after discharge and prior to interview compared with 58% of patients sent to an SNF. On multivariate analysis, mortality increased with age (confidence interval [CI] 1.04-1.09), and government insurance (CI 0.34-0.94), but decreased with discharge to home (CI 1.68-4.47). There were no differences in FIM or Short-Form-12 scores between groups. Long-term mortality after discharge in elderly burn survivors is substantial. Patients sent to an SNF or with government insurance had increased mortality postdischarge. These data suggest that issues that may influence disposition status of elderly burn patients should be optimized prior to discharge to mitigate adverse outcomes associated with SNF placement.


Toll-like Receptor (TLR) Signaling Interacts with CREBH to Modulate High-density Lipoprotein (HDL) in Response to Bacterial Endotoxin.
Bacterial endotoxin can induce inflammatory and metabolic changes in the host. In this study, we revealed a molecular mechanism by which a stress-inducible, liver-enriched transcription factor, cAMP-responsive element-binding protein hepatic-specific (CREBH), modulates lipid profiles to protect the liver from injuries upon the bacterial endotoxin lipopolysaccharide (LPS). LPS challenge can activate CREBH in mouse liver tissues in a toll-like receptor (TLR)/MyD88-dependent manner. Upon LPS challenge, CREBH interacts with TNF receptor-associated factor 6 (TRAF6), an E3 ubiquitin ligase that functions as a key mediator of TLR signaling, and this interaction relies on MyD88. Further analysis demonstrated that TRAF6 mediates K63-linked ubiquitination of CREBH to facilitate CREBH cleavage and activation. CREBH directly activates expression of the gene encoding Apolipoprotein A4 (ApoA4) under LPS challenge, leading to modulation of high-density lipoprotein (HDL) in animals. CREBH deficiency led to reduced production of circulating HDL and increased liver damage upon high-dose LPS challenge. Therefore, TLR/MyD88-dependent, TRAF6-facilitated CREBH activation represents a mammalian hepatic defense response to bacterial endotoxin by modulating HDL.


Combined effects of streptozotocin and mitozolomide against four human cell lines of the Mer+ phenotype.
The majority of human tumor cell lines are proficient in the repair of guanine O6-alkylations (designated Mer+) and are thus capable of preventing the cytotoxic effects of chloroethylating agents. It has been proposed that in these cells guanine O6-chloroethylations are rapidly removed by the enzyme O6-alkylguanine DNA alkyltransferase before the formation of DNA interstrand cross-links can occur. In this study pretreatment of four Met+ human cells (A2182 lung carcinoma, A375 melanoma, HT-29 colon carcinoma, and IMR-90 normal lung fibroblasts) with the DNA methylating agent streptozotocin apparently saturates the monoadduct repair system and allows mitozolomide to form interstrand cross-links in these cells. The inhibition of the alkyltransferase results in the continued presence of guanine O6-chloroethylations which then undergo a series of reactions that lead to DNA interstrand cross-link formation. As observed by colony forming assays, streptozotocin pretreatment causes a dramatic increase in the sensitivity of these four Mer+ cell lines to the cytotoxic effects of mitozolomide. These results indicate that a combination of streptozotocin pretreatment followed by mitozolomide may be useful in the treatment of human cancer.


Epidermal growth factor (EGF) promotes phosphorylation at threonine-654 of the EGF receptor: possible role of protein kinase C in homologous regulation of the EGF receptor.
Treatment of cells with tumor-promoting phorbol diesters, which causes activation of protein kinase C, leads to phosphorylation of the epidermal growth factor (EGF) receptor at threonine-654. Addition of phorbol diesters to intact cells causes inhibition of the EGF-induced tyrosine-protein kinase activity of the EGF receptor and it has been suggested that this effect of phorbol diesters is mediated by the phosphorylation of the receptor by protein kinase C. We measured the activity of protein kinase C in A431 cells by determining the incorporation of [32P]phosphate into peptides containing threonine-654 obtained by trypsin digestion of EGF receptors. After 3 h of exposure to serum-free medium, A431 cells had no detectable protein kinase C activity. Addition of EGF to these cells resulted in [32P] incorporation into threonine-654 as well as into tyrosine residues. This indicates that EGF promotes the activation of protein kinase C in A431 cells. The phosphorylation of threonine-654 induced by EGF was maximal after only 5 min of EGF addition and the [32P] incorporation into threonine-654 reached 50% of the [32P] in a tyrosine-containing peptide. This indicates that a significant percentage of the total EGF receptors are phosphorylated by protein kinase C. A variety of external stimuli activate Na+/H+ exchange, including EGF, phorbol diesters, and hypertonicity. To ascertain whether activation of protein kinase C is an intracellular common effector of all of these systems, we measured the activity of protein kinase C after exposure of A431 cells to hyperosmotic conditions and observed no effect on phosphorylation of threonine-654, therefore, activation of Na+/H+ exchange by hypertonic medium is independent of protein kinase C activity. Since stimulation of protein kinase C by phorbol diesters results in a decrease in EGF receptor activity, the stimulation of protein kinase C activity by addition of EGF to A431 cells contributes to a feedback mechanism which results in the attenuation of EGF receptor function.


A decade of "chromosome painting" in Lolium and Festuca.
GISH has been a particularly useful technique for studying the Lolium-Festuca species complex of forage grasses. The reason for this utility is two-fold: (i) the complex is unique amongst crop plants in which fertile hybrids, and backcross progenies, can be produced which recombine genomes and promiscuously exchange their genes through homoeologous recombination; (ii) dispersed repetitive DNAs differ between species, and this allows tracking of the identity of chromosomes and chromosome segments. This tracking property has enabled several fruitful lines of research to produce a harvest of new information for both fundamental and practical purposes. We review this first decade of GISH (genomic in situ hybridization) in Lolium-Festuca, and discuss and summarize the achievements which have accrued.


Dying at home: evaluation of a hospice rapid-response service.
Various studies suggest that there is a preference among patients, professionals and the public for death to occur at home (Dunlop et al, 1989; Townsend et al, 1990; Hinton, 1994). Data indicates that some patients are denied the opportunity to exercise choice in the place of death. In areas where palliative rapid-response teams have been available more people have been able to die at home. This article presents the findings of an evaluation of a hospice rapid-response service in the Highlands of Scotland, which was designed to respond to the needs of people in crisis facing death. The study was designed to produce a rigorous and comprehensive description of a newly-operational service at the pilot stage. It reflects not only the views of the professionals involved but also focuses on the views of informal carers. Seventeen patients were referred to the rapid-response service in its first year of operation between April 1998 and March 1999. The service has shown some success in enabling patients to die at home and satisfaction with the service is high among professionals and lay carers.


Vasectomy and prostate cancer: a case-control study in a health maintenance organization.
A case-control study was conducted within Group Health Cooperative of Puget Sound to evaluate the relation between prior vasectomy and the risk of prostate cancer. Cases consisted of 175 men newly diagnosed with histologically confirmed prostate cancer during 1989-1991. A total of 258 controls, matched to cases on birth year and membership status, were randomly selected from the general membership of the plan. Information was collected from mailed questionnaires and medical records on medical history, including prior vasectomy, anthropometric measures, family history of prostate cancer, personal habits, and medical care utilization, and demographic factors. Conditional logistic regression analyses showed that the odds ratio for prostate cancer associated with vasectomy was 0.86 (95% confidence interval 0.57-1.32) after adjustment for confounders. The odds ratio estimate did not differ substantially by age at vasectomy or time since vasectomy. However, the odds ratio estimate for prostate cancer associated with vasectomy tended to be increased among men who had a father or brother with prostate cancer. Nevertheless, the increased risk may be related to detection bias or differential participation rates due to both vasectomy status and a family history of prostate cancer. These results suggest no overall association between vasectomy and prostate cancer.


Effects of neonatal handling on play behavior and fear towards a predator odor in juvenile rats (Rattus norvegicus).
The effects of brief daily separation, also known as "handling," during the first 2 weeks of life on play behavior and fearfulness toward a predatory odor were assessed in juvenile rats. Handled rats were more playful than nonhandled control rats, and while handling had no effect on the direct response of these rats toward a predatory odor, handled rats did not exhibit a conditioned suppression of play when tested later in the same context where they had been exposed to the predatory odor. Handled rats were still wary of the environment in that they continued to show a heightened level of risk assessment behavior. These data suggest that early postnatal experiences may play a significant role in determining how an animal deals with predatory threats later in life.


Vascular endothelial growth factor directly stimulates tumour cell proliferation in non-small cell lung cancer.
Vascular endothelial growth factor (VEGF) is a key stimulator of physiological and pathological angiogenesis. VEGF signals primarily through VEGF receptor 2 (VEGFR2), a receptor tyrosine kinase whose expression is found predominantly on endothelial cells. The purpose of this study was to determine the role of VEGFR2 expression in NSCLC cells. NSCLC cells and tissue sections were stained for VEGFR2 expression by immunohistochemistry (IHC). Immunoblotting and ELISA were used to determine the activation and inhibition of VEGFR2 and its downstream signalling pathways. Five-day proliferation assays were carried out in the presence or absence of VEGF. IHC analysis of NSCLC demonstrated tumour cell VEGFR2 expression in 20% of samples. Immunoblot analysis showed expression of VEGFR2 protein in 3/8 NSCLC cell lines that correlated with VEGFR2 mRNA expression levels. VEGF-dependent VEGFR2 activation was apparent in NSCLC cells, and was associated with increased tumor cell proliferation. Cediranib treatment or siRNA against VEGFR2 inhibited VEGF-dependent increases in cell proliferation. Inhibition of VEGFR2 tyrosine kinase activity using cediranib was more effective than inhibition of AKT (MK2206) or MEK (AZD6244) for overcoming VEGFR2-driven cell proliferation. VEGF treatment did not affect cell survival following treatment with radiation, cisplatin, docetaxel or gemcitabine. Our data suggest that a subset of NSCLC tumour cells express functional VEGFR2 which can act to promote VEGF-dependent tumour cell growth. In this tumour subset, therapies targeting VEGFR2 signalling, such as cediranib, have the potential to inhibit both tumour cell proliferation and angiogenesis.


A Computational Study on Synaptic Plasticity Regulation and Information Processing in Neuron-Astrocyte Networks.
Postsynaptic ionotropic receptors critically shape synaptic currents and underpin their activity-dependent plasticity. In recent years, regulation of expression of these receptors by slow inward and outward currents mediated by gliotransmitter release from astrocytes has come under scrutiny as a potentially important mechanism for the regulation of synaptic information transfer. In this study, we consider a model of astrocyte-regulated synapses to investigate this hypothesis at the level of layered networks of interacting neurons and astrocytes. Our simulations hint that gliotransmission sustains the transfer function across layers, although it decorrelates the neuronal activity from the signal pattern. Overall, our results make clear how astrocytes could transform neuronal activity by inducing a lowfrequency modulation of postsynaptic activity.


Ethics of practice among Christian psychologists: a pilot study.
Christian psychologists have questioned whether the code of ethics proposed by secular psychology is sufficient for the practitioner who is a Christian. However, little descriptive data exist on the ethical beliefs and behaviors of Christian psychologists. Survey data were obtained from 69 Christian clinicians indicating the frequency with which they engaged in 83 behaviors and the degree to which they believed the behaviors were ethical. Responses were tabulated for each option and on each behavior. Of the 83 items, four occurred in the practices of over 90% of the respondents, while six never occurred, the latter being mostly sexually related items. Agreement on ethics was found with 13 of the items, while nine behaviors, several of which involved financial issues, proved to be difficult ethical judgments. The data indicate that Christian psychologists are effectively understanding and implementing the ethical guidelines of the American Psychological Association although guidance may be needed regarding finances and homosexuality.


Suicidal Tendency Among Adolescents With Adjustment Disorder.
Adolescents with adjustment disorder (AD) are at risk of presenting suicidal symptoms. Certain personality traits are linked to suicidal tendencies. There is a lack of information about the link between suicide and personality patterns in adolescents with AD. To identify the personality characteristics that predispose to or prevent the development of suicidal ideation and behavior among adolescents with AD. We recruited 108 adolescents with AD at a public mental health center near Barcelona (Spain). They were administered the Inventario de Riesgo Suicida para Adolescentes (IRIS) to assess suicidal symptoms, as well as the Millon Adolescent Clinical Inventory (MACI) and the 16PF Adolescent Personality Questionnaire (16PF-APQ) to appraise personality features. Doleful personality emerged as the principal risk for suicidal symptoms. The conforming personality pattern exerted a protective effect, and emotional stability was associated with low levels of suicidal tendencies. Among the Big Five factors, anxiety had the highest explanatory power for suicidal tendencies. Certain personality characteristics are associated with heightened or reduced risk of suicidal tendencies in adolescents with AD. Their identification is important for clinicians designing treatment programs for these patients.


Bio-inspired electrospun micro/nanofibers with special wettability.
Inspired by the extreme wetting states displayed by the natural materials, various techniques have been widely investigated to fabricate superhydrophobic and superhydrophilic surfaces. Electrospinning has gained huge amount of interest as fibers with suitable combination of surface chemistry and surface roughness can be easily obtained. This study provides a comprehensive overview of the progress that has been made on electrospun fibers that display superhydrophobicity, superhydrophilicity or a combination of both. The article discusses various modification techniques that can be implemented to obtain fibers with surface heterogeneity for improving its hydrophobicity or hydrophilicity. Both nanometer size of the fibers and secondary nanoscale structures ensure that the fibers have suitable surface topography to exhibit extreme wetting states. Additionally, for the first time, we critically review and identify the role of intrinsic structures such as crystallinity and chain orientation on the wettability of the fibers. We highlight some new emerging application areas that are being explored using superhydrophobic and superhydrophilic fibers. Further, methods for fabricating smart materials with special wettability are also discussed. Such fibers with special wettability show tremendous promise for water harvesting, unidirectional water collection and oil-water filtration applications.


Plasma Leak From the Circulation Contributes to Poor Outcomes for Preterm Infants: A Working Hypothesis.
Preterm infants are at high risk of death and disability resulting from brain injury. Impaired cardiovascular function leading to poor cerebral oxygenation is a significant contributor to these adverse outcomes, but current therapeutic approaches have failed to improve outcome. We have re-examined existing evidence regarding hypovolemia and have concluded that in the preterm infant loss of plasma from the circulation results in hypovolemia; and that this is a significant driver of cardiovascular instability and thus poor cerebral oxygenation. High capillary permeability, altered hydrostatic and oncotic pressure gradients, and reduced lymphatic return all combine to increase net loss of plasma from the circulation at the capillary. Evidence is presented that early hypovolemia occurs in preterm infants, and that capillary permeability and pressure gradients all change in a way that promotes rapid plasma loss at the capillary. Impaired lymph flow, inflammation and some current treatment strategies may further exacerbate this plasma loss. A framework for testing this hypothesis is presented. Understanding these mechanisms opens the way to novel treatment strategies to support cardiovascular function and cerebral oxygenation, to replace current therapies, which have been shown not to change outcomes.


Counteracting effects of dexamethasone and alpha 2-macroglobulin on inhibition of proliferation of normal and neoplastic rat hepatocytes by transforming growth factors-beta type 1 and type 2.
Primary cultures of hepatocytes isolated from normal F-344 rats or from F-344 rats with hepatocellular carcinomas generated by a 2-step model of chemical carcinogenesis were used to determine if dexamethasone (DEX) or alpha 2-macroglobulin (alpha 2M) modify the ability of transforming growth factors-beta type I (TGF-beta I) and type 2 (TGF-beta 2) to inhibit labelling index of hepatocytes cultured continuously with or without epidermal growth factor (EGF). Both TGF-beta 1 and beta 2 were equivalently potent inhibitors of S-phase DNA synthesis in normal and neoplastic hepatocytes as determined by 3H-thymidine autoradiography. Both DEX (1 to 100 microM) and alpha 2M (50-200 microM) partially counteracted the mito-inhibitory effect of both TGF-betas on the proliferation of normal and surrounding hepatocytes. In contrast, neoplastic hepatocytes cultured with DEX released much less immunoreactive alpha 2M and were less able to overcome the inhibitory effect of TGF-beta than normal or surrounding hepatocytes. Purified bovine alpha 2M partially counteracted the inhibition of TGF-beta 1 or beta 2 of both surrounding and neoplastic hepatocytes. Both DEX and alpha 2M were more effective against the mito-inhibitory activity of TGF-beta 2. Our data suggest that alpha 2M released by DEX-treated normal hepatocytes contributes to the counteraction of the TGF-beta effect by DEX. Our results support the hypothesis that glucocorticoids and growth-factor-binding proteins may have important roles in modulating the effects of TGF-beta on normal hepatocyte proliferation and suggest that under some conditions hepatocellular neoplasms can be more sensitive than normal hepatocytes to inhibition of proliferation by TGF-beta.


A brave new world for an old world pest: Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil.
The highly polyphagous Old World cotton bollworm Helicoverpa armigera is a quarantine agricultural pest for the American continents. Historically H. armigera is thought to have colonised the American continents around 1.5 to 2 million years ago, leading to the current H. zea populations on the American continents. The relatively recent species divergence history is evident in mating compatibility between H. zea and H. armigera under laboratory conditions. Despite periodic interceptions of H. armigera into North America, this pest species is not believed to have successfully established significant populations on either continent. In this study, we provide molecular evidence via mitochondrial DNA (mtDNA) cytochrome oxidase I (COI) and cytochrome b (Cyt b) partial gene sequences for the successful recent incursion of H. armigera into the New World, with individuals being detected at two sites (Primavera do Leste, Pedra Preta) within the State of Mato Grosso in Brazil. The mtDNA COI and Cyt b haplotypes detected in the Brazilian H. armigera individuals are common throughout the Old World, thus precluding identification of the founder populations. Combining the two partial mtDNA gene sequences showed that at least two matrilines are present in Brazil, while the inclusion of three nuclear DNA Exon-Primed Intron-Crossing (EPIC) markers identified a further two possible matrilines in our samples. The economic, biosecurity, resistance management, ecological and evolutionary implications of this incursion are discussed in relation to the current agricultural practices in the Americas.


Associations of adverse childhood experiences with educational attainment and adolescent health and the role of family and socioeconomic factors: A prospective cohort study in the UK.
Experiencing multiple adverse childhood experiences (ACEs) is a risk factor for many adverse outcomes. We explore associations of ACEs with educational attainment and adolescent health and the role of family and socioeconomic factors in these associations. Using data from the Avon Longitudinal Study of Parents and Children (ALSPAC), a prospective cohort of children born in southwest England in 1991-1992, we assess associations of ACEs between birth and 16 years (sexual, physical, or emotional abuse; emotional neglect; parental substance abuse; parental mental illness or suicide attempt; violence between parents; parental separation; bullying; and parental criminal conviction, with data collected on multiple occasions between birth and age 16) with educational attainment at 16 years (n = 9,959) and health at age 17 years (depression, obesity, harmful alcohol use, smoking, and illicit drug use; n = 4,917). We explore the extent to which associations are robust to adjustment for family and socioeconomic factors (home ownership, mother and partner's highest educational qualification, household social class, parity, child's ethnicity, mother's age, mother's marital status, mother's depression score at 18 and 32 weeks gestation, and mother's partner's depression score at 18 weeks gestation) and whether associations differ according to socioeconomic factors, and we estimate the proportion of adverse educational and health outcomes attributable to ACEs or family or socioeconomic measures. Among the 9,959 participants (49.5% female) included in analysis of educational outcomes, 84% reported at least one ACE, 24% reported 4 or more ACEs, and 54.5% received 5 or more General Certificates of Secondary Education (GCSEs) at grade C or above, including English and Maths. Among the 4,917 participants (50.1% female) included in analysis of health outcomes, 7.3% were obese, 8.7% had depression, 19.5% reported smoking, 16.1% reported drug use, and 10.9% reported harmful alcohol use. There were associations of ACEs with lower educational attainment and higher risk of depression, drug use, and smoking. For example, odds ratios (ORs) for 4+ ACEs compared with no ACEs after adjustment for confounders were depression, 2.4 (1.6-3.8, p < 0.001); drug use, 3.1 (2.1-4.4, p < 0.001); and smoking, 2.3 (1.7-3.1, p < 0.001). Associations with educational attainment attenuated after adjustment but remained strong; for example, the OR after adjustment for confounders for low educational attainment comparing 4+ ACEs with no ACEs was 2.0 (1.7-2.4, p < 0.001). Associations with depression, drug use, and smoking were not altered by adjustment. Associations of ACEs with harmful alcohol use and obesity were weak. For example, ORs for 4+ ACEs compared with no ACEs after adjustment for confounders were harmful alcohol use, 1.4 (0.9-2.0, p = 0.10) and obesity, 1.4 (0.9-2.2, p = 0.13) We found no evidence that socioeconomic factors modified the associations of ACEs with educational or health outcomes. Population attributable fractions (PAFs) for the adverse educational and health outcomes range from 5%-15% for 4+ ACEs and 1%-19% for low maternal education. Using data from multiple questionnaires across a long period of time enabled us to capture a detailed picture of the cohort members' experience of ACEs; however, a limitation of our study is that this resulted in a high proportion of missing data, and our analyses assume data are missing at random. This study demonstrates associations between ACEs and lower educational attainment and higher risks of depression, drug use, and smoking that remain after adjustment for family and socioeconomic factors. The low PAFs for both ACEs and socioeconomic factors imply that interventions that focus solely on ACEs or solely on socioeconomic deprivation, whilst beneficial, would miss most cases of adverse educational and health outcomes. This interpretation suggests that intervention strategies should target a wide range of relevant factors, including ACEs, socioeconomic deprivation, parental substance use, and mental health.


Synthesis and antioxidant activity of nitrohydroxytyrosol and its acyl derivatives.
A series of nitroderivatives has been synthesized from hydroxytyrosol, the natural olive oil phenol, to increase the assortment of compounds with putative effects against Parkinson's disease. Nitrohydroxytyrosyl esters were obtained from nitrohydroxytyrosol using a chemoselective one-step, high-yield, transesterification procedure. The antioxidant activity of these new series of nitrocatechols was evaluated using FRAP, ABTS, and ORAC assays and compared to that of free hydroxytyrosol. The nitro functional group induced a significant increase in the antioxidant activity of nitrohydroxytyrosol compared to hydroxytyrosol. Regarding nitroester derivatives, variable antioxidant activity was observed depending on the acyl side-chain length; shorter chains maintained or even enhanced the antioxidant activity compared to nitrohydroxytyrosol, decreasing the activity with longer side chains in keeping with their lipophilic nature. Therefore, it may be concluded that nitroester derivatives of hydroxytyrosol, which may be obtained by a simple, high-yield reaction, have elevated antioxidant activity and thus present potential bioactivity.


Development of macromolecular prodrug for rheumatoid arthritis.
Rheumatoid arthritis (RA) is a chronic autoimmune disease that is considered to be one of the major public health problems worldwide. The development of therapies that target tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and co-stimulatory pathways that regulate the immune system have revolutionized the care of patients with RA. Despite these advances, many patients continue to experience symptomatic and functional impairment. To address this issue, more recent therapies that have been developed are designed to target intracellular signaling pathways involved in immunoregulation. Though this approach has been encouraging, there have been major challenges with respect to off-target organ side effects and systemic toxicities related to the widespread distribution of these signaling pathways in multiple cell types and tissues. These limitations have led to an increasing interest in the development of strategies for the macromolecularization of anti-rheumatic drugs, which could target them to the inflamed joints. This approach enhances the efficacy of the therapeutic agent with respect to synovial inflammation, while markedly reducing non-target organ adverse side effects. In this manuscript, we provide a comprehensive overview of the rational design and optimization of macromolecular prodrugs for treatment of RA. The superior and the sustained efficacy of the prodrug may be partially attributed to their Extravasation through Leaky Vasculature and subsequent Inflammatory cell-mediated Sequestration (ELVIS) in the arthritic joints. This biologic process provides a plausible mechanism, by which macromolecular prodrugs preferentially target arthritic joints and illustrates the potential benefits of applying this therapeutic strategy to the treatment of other inflammatory diseases.


Ganglioside (GM1)-treated T cells shed CD4.
In previous studies (Morrison et al., 1990), we showed that ganglioside (GM1) modulation of CD4 was associated with activation of phospholipase C and increased production of inositol triphosphate, but not with activation of protein kinase C. These results demonstrated a unique signal transduction pathway related to GM1 modulation of CD4 on T cells and raised the question as to whether intracellular Ca2+ levels and related protein kinases would be affected by GM1-induced signalling. We now show that GM1 modulation of CD4 from human T cells corresponds to decreased cellular Ca2+ without significant changes in cellular protein phosphorylation. In the course of this study we discovered that T cells challenged with GM1 exhibited new proteins in their surrounding media. Fractionation of cellular and supernatant proteins show that cells treated with GM1 released proteins with an approximate molecular weight (Mr) of 49,000. This was exclusive of GM1 protein association and GM1-induced protein phosphorylation. Immunoblots demonstrated the presence of CD4 in GM1-treated cell supernatants. Western immunoblots using anti-CD4 antibodies detected a lower Mr form (49,000) of CD4 in the supernatants of GM1-treated cells. These studies further define the unique nature of GM1 signalling relating to modulation of CD4 and demonstrate that the fate of GM1 modulated CD4, in part, involves protein shedding.


The relationship between antidepressant medication use and rate of suicide.
Approximately 30 000 people die annually by suicide in the United States. Although 60% of suicides occur during a mood disorder, mostly untreated, little is known about the relationship between antidepressant medication use and the rate of suicide in the United States. To examine the association between antidepressant medication prescription and suicide rate by analyzing associations at the county level across the United States. Analysis of National Vital Statistics from the Centers for Disease Control and Prevention. All US counties. All US individuals who committed suicide between 1996 and 1998. National county-level suicide rate data are broken down by age, sex, income, and race for the period of 1996 to 1998. National county-level antidepressant prescription data are expressed as number of pills prescribed. The primary outcome measure is the suicide rate in each county expressed as the number of suicides for a given population size. The overall relationship between antidepressant medication prescription and suicide rate was not significant. Within individual classes of antidepressants, prescriptions for selective serotonin reuptake inhibitors (SSRIs) and other new-generation non-SSRI antidepressants (eg, nefazodone hydrochloride, mirtazapine, bupropion hydrochloride, and venlafaxine hydrochloride) are associated with lower suicide rates (both within and between counties). A positive association between tricyclic antidepressant (TCA) prescription and suicide rate was observed. Results are adjusted for age, sex, race, income, and county-to-county variability in suicide rates. Higher suicide rates in rural areas are associated with fewer antidepressant prescriptions, lower income, and relatively more prescriptions for TCAs. The aggregate nature of these observational data preclude a direct causal interpretation of the results. A high number of TCA prescriptions may be a marker for those counties with more limited access to quality mental health care and inadequate treatment and detection of depression, which in turn lead to increased suicide rates. By contrast, increases in prescriptions for SSRIs and other new-generation non-SSRIs are associated with lower suicide rates both between and within counties over time and may reflect antidepressant efficacy, compliance, a better quality of mental health care, and low toxicity in the event of a suicide attempt by overdose.


Smoking and the long-term incidence of cataract: the Blue Mountains Eye Study.
To assess the association between smoking and the long-term incidence of cataract and cataract surgery. In a population-based cohort of Australians aged 49 years and over, 3654 participants were seen at baseline (1992-94) and 2406 were seen after 5- and/or 10-years and had photographs taken to assess incident cataract. Smoking status was recorded at interview. History, physical examination and fasting blood samples provided data on possible risk factors. Cataract incidence was calculated using the Kaplan-Meier survival approach. Discrete linear logistic models were used to assess risk of incident cataract. After controlling for age, sex and other factors, ever smokers had an increased risk of developing nuclear cataract compared to never smokers (relative risk (RR) 1.41; 95% confidence interval (CI) 1.09-1.83; RR 1.37, CI 1.04-1.81 for past smoking and RR 1.57, CI 1.06-2.31 for current smoking). The effect of smoking was strongest in ever smokers reporting 36 + pack-years of smoking compared to never smokers (RR 1.46; CI, 1.02-2.08). Current smokers also developed nuclear cataract slightly younger than non-smokers (mean age 65.2 versus 67.5 years, p = 0.049). No statistically significant associations were found between smoking status and the incidence of cortical or posterior subcapsular cataract, or cataract surgery. These epidemiological data confirm smoking as a modest risk factor for the development of nuclear cataract.


CT prognostic criteria of survival after malignant glioma surgery.
The serial pre- and postoperative computed tomographic (CT) scans of 115 patients entered in the Cooperative Brain Tumor Study between 1975 and 1982 were analyzed in order to define CT prognostic criteria and to test the hypothesis that radical glioma surgery prolongs patient survival. The CT parameters of mass size, associated edema, and intensity of enhancement were quantitated on all scans. Clinical parameters evaluated included gender, age, length of survival, and useful (Karnofsky greater than 30) survival. Data analyses indicated postoperative residual tumor burden was inversely related to length of survival (p less than 0.01). Postoperative associated edema and intensity of image enhancement were also of prognostic value and showed an inverse relation to survival. Younger patients proved more likely than older patients to attain long-term survival. Residual tumor burden of less than 45 mm diameter on postoperative CT scans was associated with 70% chance of long-term survival. These findings support the radical surgical management of glioma.


The 213-amino-acid leucine-rich repeat region of the listeria monocytogenes InlB protein is sufficient for entry into mammalian cells, stimulation of PI 3-kinase and membrane ruffling.
The Listeria monocytogenes InlB protein is a 630-amino-acid surface protein that mediates entry of the bacterium into a wide variety of cell types, including hepatocytes, fibroblasts and epithelial cells such as Vero, HEp-2 and HeLa cells. Invasion stimulates host proteins tyrosine phosphorylation, PI 3-kinase activity and rearrangements in the actin cytoskeleton. We previously showed that InlB is sufficient for entry of InlB-coated latex beads into cells and recent results indicate that purified InlB can stimulate PI 3-kinase activity and is thus the first bacterial agonist of this lipid kinase. In this study, we identified the region of InlB responsible for entry and stimulation of signal transduction events. Eight monoclonal antibodies directed against InlB were raised and, of those, five inhibited bacterial entry. These five antibodies recognized epitopes within the leucine-rich repeat (LRR) region and/or the inter-repeat (IR) region. InlB-staphylococcal protein A (SPA) fusion proteins and recombinant InlB derivatives were generated and tested for their capacity to mediate entry into cultured mammalian cells. All the InlB derivatives that carried the amino-terminal 213-amino-acid LRR region conferred invasiveness to the normally non-invasive bacterium L. innocua or to inert latex beads and the corresponding purified polypeptides inhibited bacterial entry. In addition, the 213-amino-acid LRR region was able to stimulate PI 3-kinase activity and changes in the actin cytoskeleton (membrane ruffling). These properties were not detected with purified internalin, another invasion protein of L. monocytogenes that displays LRRs similar to those of InlB. Taken together, these results show that the first 213 amino acids of InlB are critical for its specific properties.


Internal standard application to dried blood spots by spraying: investigation of the internal standard distribution.
The scientifically and logistically best way of application of the internal standard (IS) in the analysis of dried blood spots (DBS) analysis is still a matter of debate and investigation. Most commonly the IS is added in the solvent used for extraction of the discs punched from DBS. In this case, the recovery of the non-extracted IS is complete while the recovery of the analyte extracted from DBS is different from the IS. An alternative way for addition of the IS was investigated. A homogeneous distribution and absorption of the test compound across the spots was demonstrated by spraying a solution of a radiolabeled test compound (mimicking an IS solution) onto DBS. This spray-on technique is convenient and easily automatable. Spraying of the solution was rapid, precise and reproducible, and therefore seems to be suitable for routine analysis of DBS by offline and online extraction.


Apoptosis and Fas/Fas ligand mRNA expression in acute immune complex alveolitis in mice.
Deoxyribonucleic acid (DNA) strand breaks as a characteristic of apoptosis, and Fas antigen (Fas)/Fas ligand (FasL) expression may participate in acute immune complex alveolitis in mice. Male Institute for Cancer Research (ICR) mice were injected intravenously with immunoglobulin G (IgG) antibodies against ovalbumin and inhaled an aerosolized oval albumin (OA) solution. They were killed at 4, 6, 12, 24, 48 h and 7 days after aerosolization. We assessed DNA fragmentation by agarose gel electrophoresis and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end-labelling (TUNEL). The expression of Fas and FasL messenger ribonucleic acid (mRNA) in lung tissues was assessed by reverse transcriptase (RT) polymerase chain reaction, and by in situ hybridization (ISH) to localize Fas mRNA, and RT in situ polymerase chain reaction to localize FasL mRNA. The fragmentation of DNA extracted from lung tissue was found 6-24 h after OA inhalation. TUNEL detected positive signals in bronchial and alveolar epithelial, endothelial and inflammatory cells in the lung tissue. These positive signals had disappeared 7 days after OA inhalation. TUNEL also detected positive signals in apoptotic neutrophils in bronchoalveolar lavage fluid at 6-12 h. Fas mRNA was expressed in the alveolar epithelial and inflammatory cells, while the expression of FasL mRNA appeared to be upregulated in infiltrating inflammatory cells at 6-24 h. These results suggest that apoptosis may be associated with the resolution of inflammation and with tissue repair and also suggest the involvement of the Fas antigen/Fas ligand pathway in acute immune complex alveolitis in mice.


Long-term administration of amnion-derived cellular cytokine suspension promotes functional recovery in a model of penetrating ballistic-like brain injury.
Previous work has shown that human amnion-derived progenitor (AMP) cell therapy is neuroprotective in a penetrating ballistic-like brain injury (PBBI) model. However, the neuroprotective capacity of AMP cells seemed to be mediated by the sustained secretion of AMP cell-derived neurotrophic factors, which are abundant in the amnion-derived cellular cytokine suspension (ACCS). To test this theory, the current study assessed the neuroprotective efficacy of long-term ACCS delivery in the PBBI model. Experiment 1 assessed the bioactive stability and neuroprotective capacity of ACCS in an in vitro model of neurodegeneration. Experiment 2 evaluated the therapeutic effects of ACCS delivery initiated 15 minutes after PBBI and continued for 2 weeks after injury. Experiment 3 was designed to identify the therapeutic window for long-term ACCS delivery in the PBBI model. Outcome metrics included neurobehavioral assessments and neuropathologic measures of neuroinflammation and axonal/neuronal degeneration. Experiment 1 demonstrated that ACCS is thermally stable for 1 week at 37°C and that ACCS treatment protected neurite against staurosporine toxicity. Experiment 2 identified the optimal infusion rate of ACCS (1 μL/h) and demonstrated that long-term infusion of ACCS was capable of promoting significant protection against PBBI-induced neuropathology and motor abnormalities, but was not sufficient for reducing cognitive deficits. Finally, the results of Experiment 3 showed that ACCS is effective in promoting significant neuroprotection even when onset of treatment is delayed out to 24 hours (but not 48 hours) after PBBI. Collectively, our results support the hypothesis that the neuroprotective effects of AMP cells are mediated through a sustained delivery of ACCS, which implicates ACCS as a promising neuroprotection agent for clinical study.


[Proteolysis in digestive system regulation].
Signal enzymes with direct and indirect hormone releasing action are formed by means of proteolysis from exogenic and endogenic proteins. The proteolysis is the basis of hormone processing. The limited proteolysis forms hormones from pro-hormones, ligand proteolysis excludes or reduces their stimulated or inhibited effects. The existence of polipotent proteinaso-activated receptors with regulative and modulated role in norm and pathology was proved.


Acute benzo[a]pyrene exposure induced oxidative stress, neurotoxicity and epigenetic change in blood clam Tegillarca granosa.
The blood clam (Tegillarca granosa) is being developed into a model bivalve mollusc for assessing and monitoring marine pollution on the offshore seabed. However, the information on the response of blood clam to PAHs, an organic pollutant usually deposited in submarine sediment, remains limited. Herein, we employed multiple biomarkers, including histological changes, oxidative stress, neurotoxicity and global DNA methylation, to investigate the effects of 10 and 100 μg/L Bap exposure on the blood clams under laboratory conditions, as well as the potential mechanisms. Acute Bap exposure can induce significant morphological abnormalities in gills as shown through hematoxylin-eosin (H.E) staining, providing an intuitive understanding on the effects of Bap on the structural organization of the blood clams. Meanwhile, the oxidative stress was significantly elevated as manifested by the increase of antioxidants activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and glutathione-s-transferase (GST), lipid peroxidation (LPO) level and 8-hydroxy-2'-deoxyguanosine (8-OHdG) content. The neurotoxicity was also strengthened by Bap toxicity manifested as inhibited acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) activities. In addition, the global DNA methylation level was investigated, and a significant DNA hypomethylation was observed in Bap exposed the blood clam. The correlation analysis showed that the global DNA methylation was negatively correlated with antioxidants (SOD, CAT and POD) activities, but positively correlated choline enzymes (AChE and ChAT) activities. These results collectively suggested that acute Bap exposure can cause damage in gills structures in the blood clam possibly by generating oxidative stress and neurotoxicity, and the global DNA methylation was inhibited to increase the transcriptional expression level of antioxidants genes and consequently elevate antioxidants activities against Bap toxicity. These results are hoped to shed some new light on the study of ecotoxicology effect of PAHs on marine bivalves.


Research on the Effect of Puerarin on Alleviating Sports Fatigue.
This paper focus on the effects of puerarin remit sports fatigue by observeing the effects of puerarinon blood components and exercise ability of mice in swimming trained. Comparing with the time of exhaustive swimming in the swim trained group, swim trained group with puerarin significantly increase (p<0.05). The puerarin may reduce the increase of hemoglobin, red blood cell and platelets caused by endurance exercise, reduce the blood viscosity, prolong the time of exhaustive swimming and improve exercise ability of mice in swim trained. Through the rat swimming experiment, we can further know the effects of puerarin on alleviating sports fatigue are obviously, which plays a role in the research field of the physiological effect of puerarin.


[Experiment in infecting the green monkey Cercopithecus aethiops with a leukocyte virus isolated in hepatitis A].
The virus isolated in hepatitis A had been previously shown to passage in primary phytohemagglutinin-stimulated leukocyte cultures. The virus designated as leukocyte hepatitis virus (LHV) causes chromosome aberrations in the infected cultures. Some physico-chemical properties of LHV and its capacity for reproduction in other cell systems were studied. In serological surveys of patients and in studies of cellular immunity reactions of them LHV was found in cases of hepatitis. The possibility of producing a laboratory model of hepatitis in green monkeys, morphological reaction to inoculation with LHV, as well as the presence of specific antigen in the inoculated animals by immunofluorescence test using rabbit immune serum to LHV and FITC-labelled gamma globulins from hepatitis A convalescents were investigated. LHV antigen was found in the lymph nodes, spleen and liver. Active morphological reaction of lymph nodes and spleen was observed 30 days after inoculation of monkeys. The results indicate that in green monkeys LHV possesses marked reticulotropic properties without producing clinical hepatitis, therefore African grivet monkeys (Cercopithecus aethiops), apparently, cannot be used as experimental models for LHV studies in vivo.


Noninvasive measurement of systolic and diastolic blood pressure in swine.
Arterial blood pressure was measured noninvasively using Doppler ultrasound and an occluding cuff. The subjects were 28 domestic pigs (10-49 kg) anesthetized with pentobarbital sodium (25 mg/kg). Indirect pressure measurements were made with the Doppler unit placed over the radial or the ulnar artery proximal to the carpal joint. Comparison was made with directly measured pressure to determine the reliability and reproducibility of the indirect method. Direct systolic pressures between 73 and 230 mmHg and diastolic pressures between 52 and 165 mmHg were measured. There was no significant difference between directly and indirectly measured systolic pressure (P greater than 0.20). Indirectly measured diastolic pressure tended to be lower than direct diastolic pressure, the difference by the paired t test being significant to P = 0.06. For systolic pressure the sample correlation coefficient was 0.94, and for diastolic pressure, 0.88. Both systolic and diastolic blood pressure can be accurately measured in the anesthetized pig using Doppler ultrasound and an occluding cuff.