Beta1-adrenergic blockade augments pulsatile PTH secretion in humans.
Pulsatile peptide hormone secretion provides efficient control of specific end organ functions. To test the hypothesis that sympathetic neuronal activity drives synchronous pulsatile PTH release from the parathyroid glands, we investigated the acute effects of beta1-adrenergic receptor blockade on PTH secretion patterns in a single-blinded study in nine healthy adults. Plasma PTH levels were determined at 1-min intervals. After a 75-min baseline period, seven subjects received a continuous intravenous infusion of the short-acting beta1-adrenergic receptor blocker esmolol for 105 min. After a 30-min washout period, esmolol was infused for another 30 min. Two additional subjects were randomized to receive solvent infusions. PTH secretion characteristics were analyzed by multiparameter deconvolution analysis, and the orderliness of plasma PTH fluctuations by Approximate Entropy statistics. BP slightly decreased during esmolol infusion, whereas heart rate, ionized calcium, phosphate, magnesium, and plasma and urine catecholamines remained unchanged. Esmolol increased mean plasma PTH by 33 +/- 8% (P < 0.01), due to a preferential increase in the pulsatile PTH secretion component (+129 +/- 44%, P < 0.02). The increased pulsatile PTH secretion was mediated by an augmented PTH burst mass (+117 +/- 42%, P < 0.01), whereas burst frequency remained unchanged. The regularity of PTH fluctuations was not affected by the beta-adrenergic blockade. The effects were reproducible during the second esmolol infusion. The authors conclude that the sympathetic nervous system has a modulating effect on pulsatile PTH secretion. Selective beta-1 adrenergic blockade acutely increases plasma PTH by augmenting the mass of hormone secreted per burst, but it does not alter the rhythmicity of pulsatile PTH release.


Accuracy of death certificates in COPD: analysis from the TORCH trial.
The Towards a Revolution in COPD Health (TORCH) trial was an international clinical trial of chronic obstructive pulmonary disease (COPD) patients where cause of death was assigned by an independent committee. Comparison of death certificate data and adjudicated cause of death allows a unique opportunity to determine death certificate accuracy and frequency of COPD listing on death certificates of COPD patients. In this analysis, the authors determine the concordance between adjudicated cause of death and primary and secondary cause of death from death certificates. In 317 (80%) of informative deaths, the primary or secondary cause of death from certificates agreed with adjudicated cause of death. Only 229 (58%) of death certificates in these COPD patients listed COPD on the certificate. COPD was not listed on the death certificate in 21% of deaths adjudicated to be caused by COPD exacerbation. Compared with pulmonary causes, the listing of COPD on certificates occurred with less frequency than cardiovascular, cancer and other categories of death. The combined primary and secondary listing on death certificates has good concordance with actual cause of death. COPD is under-reported on death certificates, and this under-reporting is more frequent when the primary cause of death is not pulmonary.


Functional steadiness of the cerebral circulatory system under altered gravitational conditions.
The maintenance of hemodynamic stability is a functional characteristic of the cerebral circulatory system. This characteristic developed during the evolutionary process. It can be of use as an indicator of the activity of the central processes regulating cerebral blood flow under different stress conditions, particularly, during a long space mission. Rheoencephalographic methods are qualitative and yield information that permits assessment of the changes in the functional status of the cerebral circulatory system. This was confirmed by results obtained during the Salut-4 space mission. The observations permitted us to assess indirectly the status of the cosmonauts.


Loneliness, Social Support Level, Quality of Life and Symptom Management Among Turkish Oncology Patients.
This cross-sectional and correlational study was performed in order to determine the relationships between the perceived loneliness and social support levels of Turkish oncology patients, as well as their quality of life and symptom management. A total of 370 oncology inpatients participated in this study. Data were collected using, the FACT-G Quality of Life Scale, the Multi-Dimensional Scale of Perceived Social Support (MSPSS), the UCLA-Loneliness Scale (UCLA-LS), and the Edmonton Symptom Assessment Scale. A negative advanced significant relationship was found between the MSPSS total scores (r = -0.754, p < 0.01) and the UCLA-LS total scores. As the social support levels of the patients increased, their loneliness levels were seen to decrease and their quality of life was seen to increase. The patients were found to experience the symptoms of fatigue, worry, and feeling unwell more often as their loneliness levels increased and social support levels decreased.


Splenic torsion after congenital diaphragmatic hernia repair: case report and review of the literature.
Wandering spleen with torsion, a rare clinical diagnosis, was found to be the cause of chronic abdominal pain in an 11-year-old female with a history of congenital diaphragmatic hernia repaired at three days of age. Doppler ultrasound revealed patent vessels with splenomegaly, and computed tomography (CT) showed an absence of the spleen in the left subphrenic space with torsion at the splenic hilum. Due to the chronicity of pain and risk of ischemia from torsion, open splenopexy with Vicryl mesh was performed. This case report/review of the literature discusses the rarity of this condition, and the importance of timely diagnosis and intervention.


Critical health literacy in American deaf college students.
This study investigates the relationship between critical health literacy (CHL) and discussion of health information among college deaf students who use American Sign Language. CHL is crucial in making appropriate health-related decisions for oneself and aiding others in making good health-choices. Research on general youth population shows that frequent health-related discussions with both friends and family is associated with higher health literacy. However, for our sample of deaf college-aged students who might have had less access to communication at home, we hypothesize that health-related discussions with same-age peers may be more important for critical health literacy. We asked two questions to assess the frequency of health-related discussions with friends and families: "How often do you discuss health-related information with your friends" and "How often do you discuss your family medical history with your family?". Participants rated their experience on a scale from 1-5 (1=never, 5=always). To assess CHL, 38 deaf and 38 hearing participants were shown a short scenario that showed a woman confiding in her friend after finding a lump in her breast. Participants were then asked what the friend should say. Responses were scored by a team of 3 raters using a CHL rubric. As predicted, results showed a strong relationship between discussion of health-related information with friends and CHL in both deaf and hearing samples. Discussion with family was linked to CHL only for hearing participants, but not deaf participants in our study. These findings underscore the importance of socializing with health-literate, accessible peers to improve the health literacy and health outcomes of all deaf people.


Expression of adhesion and extracellular matrix genes in human blastocysts upon attachment in a 2D co-culture system.
What are the changes in human embryos, in terms of morphology and gene expression, upon attachment to endometrial epithelial cells? Apposition and adhesion of human blastocysts to endometrial epithelial cells are predominantly initiated at the embryonic pole and these steps are associated with changes in expression of adhesion and extracellular matrix (ECM) genes in the embryo. Both human and murine embryos have been co-cultured with Ishikawa cells, although embryonic gene expression associated with attachment has not yet been investigated in an in vitro implantation model. Vitrified human blastocysts were warmed and co-cultured for up to 48 h with Ishikawa cells, a model cell line for receptive endometrial epithelium. Six days post-fertilization (6dpf) human embryos were co-cultured with Ishikawa cells for 12, 24 (7dpf) or 48 h (8dpf) and attachment rate and morphological development investigated. Expression of 84 adhesion and ECM genes was analysed by quantitative PCR. Immunofluorescence microscopy was used to assess the expression of three informative genes at the protein level. Data are reported on 145 human embryos. Mann-Whitney U was used for statistical analysis between two groups, with P < 0.05 considered significant. The majority of embryos attached to Ishikawa cells at the level of the polar trophectoderm; 41% of co-cultured embryos were loosely attached after 12 h and 86% firmly attached after 24 h. Outgrowth of hCG-positive embryonic cells at 8dpf indicated differentiation of trophectoderm into invasive syncytiotrophoblast. Gene expression analysis was performed on loosely attached and unattached embryos co-cultured with Ishikawa cells for 12 h. In contrast to unattached embryos, loosely attached embryos expressed THBS1, TNC, COL12A1, CTNND2, ITGA3, ITGAV and LAMA3 and had significantly higher CD44 and TIMP1 transcript levels (P = 0.014 and P = 0.029, respectively). LAMA3, THBS1 and TNC expressions were validated at the protein level in firmly attached 7dpf embryos. Thrombospondin 1 (THBS1) resided in the cytoplasm of embryonic cells whereas laminin subunit alpha 3 (LAMA3) and tenascin C (TNC) were expressed on the cell surface of trophectoderm cells. Incubation with a neutralizing TNC antibody did not affect the rate of embryo attachment or hCG secretion. None. This in vitro study made use of an endometrial adenocarcinoma cell line to mimic receptive luminal epithelium. Also, the number of embryos was limited. Contamination of recovered embryos with Ishikawa cells was unlikely based on their differential gene expression profiles. Taken together, we provide a 'proof of concept' that initiation of the implantation process coincides with the induction of specific embryonic genes. Genome-wide expression profiling of a larger sample set may provide insights into the molecular embryonic pathways underlying successful or failed implantation. A.A. was supported by a grant from the 'Instituut voor Innovatie door Wetenschap en Technologie' (IWT, 121716, Flanders, Belgium). This work was supported by the 'Wetenschappelijk Fonds Willy Gepts' (WFWG G142 and G170, Universitair Ziekenhuis Brussel). The authors declare no conflict of interest.


Diminished nap effects on memory consolidation are seen under oral contraceptive use.
Many young females take exogenous hormones as oral contraceptive (OC), a condition rarely controlled for in studies on sleep and memory consolidation even though sex hormones influence consolidation. This study investigated the effects of OCs on sleep-related consolidation of a motor and declarative task, utilizing a daytime nap protocol. Fifteen healthy, young females taking OCs came to the sleep lab for three different conditions: nap with previous learning, wake with previous learning and nap without learning. They underwent each condition twice, once during the "pill-active" weeks and once during the "pill-free" week, resulting in 6 visits. In all conditions, participants showed a significant off-line consolidation effect, independent of pill week or nap/wake condition. There were no significant differences in sleep stage duration, spindle activity or spectral EEG frequency bands between naps with or without the learning condition. The present data showed a significant off-line enhancement in memory irrespective of potential beneficial effects of a nap. In comparison to previous studies, this may suggest that the use of OCs may enhance off-line memory consolidation in motor and verbal tasks per se. These results stress the importance to control for the use of OCs in studies focusing on memory performance.


Zika virus, a cause of fever in Central Java, Indonesia.
In 1977 and 1978 selected in-patients at the Tegalyoso Hospital, Klaten, Indonesia who had recent onsets of acute fever were serologically studied for evidence for alphavirus and flavivirus infections. A brief clinical history was taken and a check list of signs and symptoms was completed on admission. Acute and convalescent phase sera from 30 patients who showed evidence that a flavivirus had caused their illnesses were tested for neutralizing antibodies to several flaviviruses which occur in South-east Asia. Paired sera from seven patients demonstrated a fourfold rise in antibody titre from acute to convalescent phase. The most common clinical manifestations observed in this series of patients included high fever, malaise, stomach ache, dizziness and anorexia. None of the seven patients had headache or rash despite the fact that headache and rash had been associated with two of the three previously studied. The onsets of illness clustered toward the end of the rainy season when populations of Aedes aegypti, a probable vector in Malaysia, were most abundant.


[Anti-smooth muscle antibodies in a hospital population].
A total of 139 sera being 60 patients with liver disease were tested for auto-antibodies. Twenty six of the 60 patients had active chronic hepatitis, 19 acute hepatitis, 3 cirrhosis, 3 hepatosplenic schistosomiasis mansoni, and 9 miscelaneous liver pathology. We found positivity for smooth muscle antibody in 26 cases: smooth muscle antibody-V in 15 cases, for smooth muscle antibody-T in 9, and for smooth muscle antibody-G in 10 other patients. Nine out of 10 patients with positive smooth muscle antibody-G had chronic active hepatitis and the remaining had cirrhosis. The other kinds of smooth muscle antibodies were irregularly distributed among the different hepatic disease studied. The higher levels of auto-antibodies were found mostly in chronic hepatic disease.


Problems in home monitoring of blood glucose.
Two hundred and forty blood glucose readings were made on 5 Glucochek machines by 2 different operators. The results on all machines correlated poorly with equivalent results from an automatic glucose analyser with major discrepancies at all glucose concentrations. There was a wide gap in performance between the 2 operators. Analysis of variance suggested that errors were not only due to inter-operator and inter-machine differences. Possible sources of error were shown to include time of reaction and the amount of blood on Dextrostix. Dextrostix colour changes measured by eye were as accurate as those by Glucochek without the rogue values found at higher glucose concentrations with Glucochek.


The MHC-II transactivator CIITA inhibits Tat function and HIV-1 replication in human myeloid cells.
We previously demonstrated that the HLA class II transactivator CIITA inhibits HIV-1 replication in T cells by competing with the viral transactivator Tat for the binding to Cyclin T1 subunit of the P-TEFb complex. Here, we analyzed the anti-viral function of CIITA in myeloid cells, another relevant HIV-1 target cell type. We sinvestigated clones of the U937 promonocytic cell line, either permissive (Plus) or non-permissive (Minus) to HIV-1 replication. This different phenotype has been associated with the expression of TRIM22 in U937 Minus but not in Plus cells. U937 Plus cells stably expressing CIITA were generated and HLA-II positive clones were selected by cell sorting and cloning. HLA and CIITA proteins were analyzed by cytofluorometry and western blotting, respectively. HLA-II DR and CIITA mRNAs were quantified by qRT-PCR. Tat-dependent transactivation was assessed by performing the HIV-1 LTR luciferase gene reporter assay. Cells were infected with HIV-1 and viral replication was evaluated by measuring the RT activity in culture supernatants. CIITA was expressed only in HLA-II-positive U937 Minus cells, and this was strictly correlated with inhibition of Tat-dependent HIV-1 LTR transactivation in Minus but not in Plus cells. Overexpression of CIITA in Plus cells restored the suppression of Tat transactivation, confirming the inhibitory role of CIITA. Importantly, HIV-1 replication was significantly reduced in Plus-CIITA cells with respect to Plus parental cells. This effect was independent of TRIM22 as CIITA did not induce TRIM22 expression in Plus-CIITA cells. U937 Plus and Minus cells represent an interesting model to study the role of CIITA in HIV-1 restriction in the monocytic/macrophage cell lineage. The differential expression of CIITA in CIITA-negative Plus and CIITA-positive Minus cells correlated with their capacity to support or not HIV-1 replication, respectively. In Minus cells CIITA targeted the viral transactivator Tat to inhibit HIV-1 replication. The generation of Plus-CIITA cells was instrumental to demonstrate the specific contribution of CIITA in terms of inhibition of Tat activity and HIV-1 restriction, independently from other cellular factors, including TRIM22. Thus, CIITA acts as a general restriction factor against HIV-1 not only in T cells but also in myeloid cells.


c-Src control of chloride channel support for osteoclast HCl transport and bone resorption.
Bone degradation by osteoclasts depends upon active transport of hydrogen ions to solubilize bone mineral. This transport is supported by the parallel actions of a proton ATPase and a chloride channel located in the osteoclast ruffled membrane. We have previously identified a novel chloride channel, p62, which appears to be the avian counterpart to CLIC-5b and is expressed coincident with the appearance of acid secretion as avian osteoclasts differentiate in culture. In this article, we show that suppression of CLIC-5b in differentiating avian osteoclasts results in decreased acidification by vesicles derived from these cells and decreased ability of the cells to resorb bone. Acidification is rescued by the presence of valinomycin, consistent with a selective loss of chloride channel but not proton pump activity. Osteoclast bone resorption is known to be dependent on the expression of the tyrosine kinase, c-Src. We show that CLIC-5b from osteoclasts has affinity for both Src SH2 and SH3 domains. We find that suppression of expression of Src in developing osteoclasts results in decreased vesicular acidification, which is rescued by valinomycin, consistent with the loss of chloride conductance in the proton pump-containing vesicles. Suppression of c-Src causes no change in the steady state level of CLIC-5b expression, but does result in failure of proton pump and CLIC-5b to colocalize in cultured osteoclast precursors. We conclude that suppression of c-Src interferes with osteoclast bone resorption by disrupting functional co-localization of proton pump and CLIC-5b.


Internal redistribution of radiolabelled silver among tissues of rainbow trout (Oncorhynchus mykiss) and European eel (Anguilla anguilla): the influence of silver speciation.
Influence of water Ag(I) speciation on pharmacokinetics of Ag(I) during a post-exposure period was investigated in rainbow trout (Oncorhynchus mykiss) and European eel (Anguilla anguilla). The rainbow trout is sensitive to waterborne ionic Ag(+) whereas the eel is tolerant. The fish were acclimated to either of two chloride concentrations, 10 or 1200 microM, in synthetic soft water and then exposed to a sublethal 24-h pulse of 1.3 microgl(-1) of 110mAg(I) added as 110mAgNO(3) in these waters. The protocol provided exposures to mainly the free ion Ag(+) (low chloride water) or mainly AgCl(aq) (high chloride water). Contents and concentrations of 110mAg(I) in tissues and body fluids were then monitored over a 67-day post-exposure period in Ag(I)-free water of the same chloride levels. Changing the speciation of Ag(I) in the water had no effect on the whole body load of 110mAg(I), but did result in differences in internal distribution. In trout, changing water Ag(I) speciation significantly altered elimination or accumulation of Ag(I) in several body compartments. Notably, trout exposed to AgCl(aq) eliminated 110mAg(I) from the kidney more quickly than trout exposed to Ag(I) primarily as Ag(+). This elimination was matched by higher concentrations of 110mAg in liver of trout exposed to Ag(I) primarily as AgCl(aq). In eel, shifting speciation from Ag(+) to AgCl(aq) hastened elimination of 110mAg(I) from mid and posterior intestine and increased 110mAg(I) retention in kidney. While there was no difference between the two fish species in whole body 110mAg(I) load, most internal body compartments of trout had higher 110mAg(I) concentrations than those in eel early in the experiment. Because tissue-specific elimination times were longer in eel than in trout, these differences were generally cancelled by the end of the 67-day depuration period. The only exception was the liver, which in trout continued to accumulate 110mAg(I) throughout the experiment but in eel remained unchanged. The combined effect of 110mAg(I) movements in the two species was that trout retained all their accumulated 110mAg(I) through the 67-day period, whereas the body burden of 110mAg(I) in eel was reduced to half initial values by day 67.


Alpha9 and beta8 integrin expression correlates with the merger of the developing mouse eyelids.
As previously reported, alpha9 integrin is expressed between the merged or fused eyelids of mice at birth, and changes in alpha9 localization occur during lid opening. To determine whether alpha9 and/or additional integrin subunits mediate the emergence and temporary fusion of the eyelids, immunofluorescence and confocal microscopy were used to evaluate the localization of various integrin subunits in the developing ocular surface of the mouse. No detectable beta5, beta6, or beta7 integrins were observed on the epithelia of the ocular surface. alpha2, alpha3, alphav, and beta1 integrins were most abundant in the basal cells beginning at 13.5 days post conception and remained primarily localized to the basal cell layers throughout development. beta4 was localized at the basal surface of the epidermal basal cells beginning at 13.5 days post conception but was not found on the corneal epithelial basal cells until after birth. alpha9 and beta8 integrins were present on suprabasal cells of the epidermis at the leading edge of the eyelid before merger and on the epithelial bridge that forms immediately after these tissues merge, suggesting that they play a role in the initial fusion of the epithelial tissues of the eyelid and in stabilizing the epithelial junction. After birth and into adulthood, beta8 was retained within the suprabasal cell layers of the epidermis, whereas alpha9 became localized to the basal cells of the epidermis, the conjunctiva, and the limbus. The lack of co-localization of beta4 with either alpha9 or beta8 in double-labeling studies suggests that alpha9 and beta8 are restricted to the lateral and apical aspects of those cells in which they are expressed. The presence of tenascin-C and laminin-5 at the epithelial junction site suggests that alpha9: tenascin-C and beta4: laminin-5 interactions may play a role in stabilizing the fusion between lids early on but do not appear to be involved in the movement of the lids across the cornea. The data presented identify specific integrins and matrix proteins that are likely to mediate eyelid fusion.


[Effects of ocean acidification on growth, phosphate and nitrate uptake of macroalgae].
The ocean acidification caused by elevated CO2 concentration can affect the growth, physiology and ecology, and nutrient uptake of macroalgae. In this paper, the growth and nutrient (PO4(3-) and NO3(-)) uptake of three species of macroalgae [Ulva pertusa, Scytosiphon lomentaria and Corallina pilulifera (calcified algae)] at three pHs (8.2, 7.9 and 7.6) were investigated under conditions of monoculture and mixed culture. Under the condition of monoculture, the percents of increased wet weights of U. pertusa, S. lomentaria and C. pilulifera on day 10 were the highest at the pHs of 7.9, 7.6 and 8.2, respectively, when compared with those on day 0, and the relative growth rates of C. pilulifera at pH 7.6 were significantly lower than those at pH 8.2. The data of mixed culture experiments suggested that lower pH was beneficial for the growth of S. lomentaria, while higher pH was beneficial for the growth of C. pilulifera. In both monoculture and mixed culture, the PO4(3-) and NO3(-) concentrations at the three pHs decreased with the time. The PO4(3-) concentrations decreased sharply by 71.9% - 99.0% from day 0 to day 2, and then decreased smoothly. Under the conditions of monoculture, the PO4(3-) uptake rates of U. pertusa, S. lomentaria and C. pilulifera were the highest at pHs of 8.2, 8.2 and 7.6, respectively. The NO3(-) uptake rates of U. pertusa and C. pilulifera in monoculture were the highest at pHs of 8.2 and 7.6, respectively. Under the conditions of mixed culture, the PO4(3-) uptake rates of U. pertusa + S. lomentaria, U. pertusa + C. pilulifera, S. lomentaria + C. pilulifera were the highest at pHs of 7.6, 8.2 and 8.2, respectively. The NO3(-) uptake rates of U. pertusa + C. pilulifera at pH 7.6 were the highest among the three pHs. The lower growth and higher nutrient (PO4(3-) and NO3(-)) uptake rates of monoculture C. pilulifera at pH 7.6 showed that the uptake and assimilation of PO4(3-) and NO3(-) were not coupled. The species composition of algae was changed due to ocean acidification although the dominant species was not change. Therefore, the different responses of growth and nutrient uptake of macroalgae to long-term ocean acidification in natural environment might lead to the change in macroalgal community.


Distribution of acetylated alpha-tubulin in Physarum polycephalum.
The expression and cytological distribution of acetylated alpha-tubulin was investigated in Physarum polycephalum. A monoclonal antibody specific for acetylated alpha-tubulin, 6-11B-1 (Piperno, G., and M. T. Fuller, 1985, J. Cell Biol., 101:2085-2094), was used to screen for this protein during three different stages of the Physarum life cycle--the amoeba, the flagellate, and the plasmodium. Western blots of two-dimensional gels of amoebal and flagellate proteins reveal that this antibody recognizes the alpha 3 tubulin isotype, which was previously shown to be formed by posttranslational modification (Green, L. L., and W. F. Dove, 1984, Mol. Cell. Biol., 4:1706-1711). Double-label immunofluorescence demonstrates that, in the flagellate, acetylated alpha-tubulin is localized in the flagella and flagellar cone. Similar experiments with amoebae interestingly reveal that only within the microtubule organizing center (MTOC) are there detectable amounts of acetylated alpha-tubulin. In contrast, the plasmodial stage gives no evidence for acetylated alpha-tubulin by Western blotting or by immunofluorescence.


Strategies for managing marine disease.
The incidence of emerging infectious diseases (EIDs) has increased in wildlife populations in recent years and is expected to continue to increase with global environmental change. Marine diseases are relatively understudied compared with terrestrial diseases but warrant parallel attention as they can disrupt ecosystems, cause economic loss, and threaten human livelihoods. Although there are many existing tools to combat the direct and indirect consequences of EIDs, these management strategies are often insufficient or ineffective in marine habitats compared with their terrestrial counterparts, often due to fundamental differences between marine and terrestrial systems. Here, we first illustrate how the marine environment and marine organism life histories present challenges and opportunities for wildlife disease management. We then assess the application of common disease management strategies to marine versus terrestrial systems to identify those that may be most effective for marine disease outbreak prevention, response, and recovery. Finally, we recommend multiple actions that will enable more successful management of marine wildlife disease emergencies in the future. These include prioritizing marine disease research and understanding its links to climate change, improving marine ecosystem health, forming better monitoring and response networks, developing marine veterinary medicine programs, and enacting policy that addresses marine and other wildlife diseases. Overall, we encourage a more proactive rather than reactive approach to marine wildlife disease management and emphasize that multidisciplinary collaborations are crucial to managing marine wildlife health.


[Characteristics of the diagnosis of purulent pyelonephritis].
Results of diagnosing pyelonephritis in 106 patients using ultrasonography, CT and thermovision were analyzed. Purulent pyelonephritis (PP) was diagnosed in 57 patients, serous pyelonephritis--in 49 patients. The sensitivity of ultrasonography in the diagnosing of PP was more than 80%, that of CT--90%. The methods used allowed the 100% improvement of the differential diagnosis of renal abscess, apostematous nephritis and pyonephrosis. The greatest problems were met in the diagnosis of renal carbuncle which was detected in 50% of cases by ultrasonography and in 54% of patients by CT. The minimum sizes of the destruction focus when the diagnosis could be reliable were 2 cm for ultrasonography and 1.5 cm for CT. Only using the findings of CT could diagnose emphysematous pyelonephritis. Thermovision could not find any definite criteria of PP. The timely diagnosis of PP allowed the organ-saving operations to be fulfilled in the overwhelming amount of the patients. Positive results of the treatment using the above mentioned diagnostic methods were obtained in 98.8% of the patients.


Are the Effects of Independent Biophysical Factors Linearly Additive? A 3D Tumor Migration Model.
Interstitial fluid flow plays a critical role in tumor cell invasion, yet this role has not been explored extensively in combination with other microenvironmental factors. Here, we establish a novel computational model of three-dimensional breast cancer cell migration to unveil the effect of interstitial fluid flow in the dependence of various extracellular matrix (ECM) physical properties. Our model integrates several principal factors: fluid dynamics, autologous chemotaxis, collagen fiber network structure, ECM stiffness, and cell-fiber and cell-flow interaction. First, independently with an aligned collagen fiber network and interstitial fluid flow, this model is validated by successfully reproducing the cell migration patterns. In the model, the interstitial fluid flow leads to directional symmetry breaking of chemotactic migration and synergizes with the ECM orientation to regulate cell migration. This synergy is universal in both the mesenchymal and the amoeboid migration modes, despite the fact that the cell-ECM interaction are different. Consequently, we construct a cell displacement function depending on these factors. Our cell migration model enables three-dimensional cancer migration prediction, mechanism exploration, and inhibition treatment design in a complex tumor microenvironment.


Novel difference in IF1 reactivity to Zn2+ in rabbit versus rat cardiomyocytes, mitochondria, and submitochondrial particles.
Zn2+ has a paradoxical effect on IF1-ATPase interaction in cardiac muscle mitochondria in so-called slow heart-rate mammalian species like rabbit. In such slow heart-rate mammalian species, it completely prevents IF1-mediated ATPase inhibition regardless of pH while concomitantly causing full IF1 binding to the ATPase, again, regardless of pH (Rouslin et al. (1993) J. Bioenerget. Biomembr. 25, 297-306). While our earlier study suggested that there are two kinds of IF1-ATPase interaction, a docking interaction and an ATPase inhibitory interaction with Zn2+ promoting docking and interfering with inhibition, it did not yield information on whether Zn2+ interacted primarily with IF1, with the ATPase, or with both. In the present study we show that, in contrast to its effects in rabbit cardiomyocytes, mitochondria, and SMP in which Zn2+ fully blocked IF1-mediated ATPase inhibition, Zn2+ actually enhanced ATPase inhibition in rat cardiomyocytes, although the extent of this effect was limited by the low level of IF1 in rat cardiomyocytes. Moreover, Zn2+ had no effect on IF1-mediated ATPase inhibition in rat heart mitochondria and, as suggested by inter and intra-species IF1 binding to SMP, the different effects of Zn2+ in rabbit versus those in rat appear to be mediated primarily through the different reactivities of rabbit and rat IF1 to Zn2+.


The packaging signal of simian immunodeficiency virus is upstream of the major splice donor at a distance from the RNA cap site similar to that of human immunodeficiency virus types 1 and 2.
Deletion mutation of the RNA 5' leader sequence of simian immunodeficiency virus (SIV) was used to localize the virus packaging signal. Deletion of sequences upstream of the major splice donor (SD) site produced a phenotype most consistent with a packaging defect when analysed by both RNase protection assay and RT-PCR. Sequences downstream of the SD were deleted and produced varying effects but did not affect packaging: a large downstream deletion had little effect on function, whereas a nested deletion produced a profound replication defect characterized by reduced protein production. Secondary structure analysis provided a potential explanation for this. The major packaging signal of SIV appears to be upstream of the SD in a region similar to that of human immunodeficiency virus type 2 (HIV-2) but unlike that of HIV-1; however, the packaging signal of all three viruses are at a similar distance from their respective cap sites. This conserved positioning suggests that it is more important in the virus life cycle than the position of the signal relative to the SD.


[Clinical experience of percutaneous endoscopic gastrostomy, jejunostomy, duodenostomy in 120 patients].
To report clinical experience of percutaneous endoscopic gastrostomy, duodenostomy, jejunostomy in 120 patients, focusing on its technique and indications. One hundred and twenty patients received percutaneous endoscopic gastrostomy, duodenostomy, jejunostomy from May 2001 to April 2004, including 75 percutaneous endoscopic gastrostomy (PEG), 42 percutaneous endoscopic jejunostomy (PEJ), 2 percutaneous endoscopic duodenostomy (PED), 1 direct percutaneous endoscopic jejunostomy (DPEJ). All tubes established by traditional pull technique. The average duration of PEG was (9 +/- 4) min, PEJ (17 +/- 6) min, DPEJ 20 min, and PED was 10 and 12 min for 2 patients, respectively. Success rate of the technique was 98.4% (120/122). Major complication rate was 0.8% (1/120), and minor complication rate was 7.5% (9/120). Clinical indications: PEG, PED and PEJ were applied for long-term enteral nutritional support in 88 patients, gastrointestinal decompression in 25 patients, and transfusing external drainage bile to gastrointestinal tract in 5 patients. Two radiation enteritis patients used PEG for gastrointestinal decompression preoperatively and long-term enteral nutritional support postoperatively. PEG, PED PEJ and DPEJ are easily handled, effective and safe, and may be widely used in clinical practice.


Aspergillus flavus: an emerging non-fumigatus Aspergillus species of significance.
Invasive aspergillosis is rare in immunocompetent people but contributes to significant morbidity and mortality in immunosuppressed patients. The majority (approximately 80%) of invasive Aspergillus infections is caused by Aspergillus fumigatus. The second most frequent (approximately 15-20%) pathogenic species is Aspergillus flavus and to a lesser extent, Aspergillus niger and Aspergillus terreus. Aspergillus flavus has emerged as a predominant pathogen in patients with fungal sinusitis and fungal keratitis in several institutions worldwide. To date, there has not been any publication exclusively reviewing the topic of A. flavus in the literature. This article reviews the microbiology, toxigenicity and epidemiology of A. flavus as well as describes the clinical characteristics, diagnosis and management of infections caused by this organism.


A phase 1 trial of nebulised heparin in acute lung injury.
Animal studies of acute lung injury (ALI) suggest nebulised heparin may limit damage from fibrin deposition in the alveolar space and microcirculation. No human studies have been undertaken to date. We assessed the feasibility, safety and potential anticoagulant effects of administration of nebulised heparin to patients with ALI. An open label phase 1 trial of four escalating doses of nebulised heparin was performed. A total of 16 ventilated patients with ALI were studied. The first group was administered a total of 50,000 U/day, the second group 100,000 U/day, the third group 200,000 U/day and the fourth group 400,000 U/day. Assessments of lung function included the PaO2/FiO2 ratio, lung compliance and the alveolar dead space fraction. Monitoring of anticoagulation included the activated partial thromboplastin time (APTT) and the thrombin clotting time. Bronchoalveolar lavage fluid was collected and the prothrombin fragment and tissue plasminogen activator levels were assessed. Analysis of variance was used to compare the effects of dose. No serious adverse events occurred for any dose. The changes over time for the PaO2/FiO2 ratio, lung compliance and the alveolar dead space fraction levels were similar for all doses. A trend to increased APTT and thrombin clotting time levels was present with higher doses (P = 0.09 and P = 0.1, respectively). For the highest dose, the APTT reached 64 seconds; following cessation of nebulised heparin, the APTT fell to 39 seconds (P = 0.06). In bronchoalveolar lavage samples a trend to reduced prothrombin fragment levels was present with higher doses (P = 0.1), while tissue plasminogen activator levels were similar for all doses. Administration of nebulised heparin to mechanically ventilated patients with ALI is feasible. Nebulised heparin was not associated with any serious adverse events, and at higher doses it increased APTT levels. Larger trials are required to further investigate the safety and efficacy of nebulised heparin. In these trials due consideration must be given to systemic anticoagulant effects. Australian Clinical trials registry ACTRN12606000388516.


Neurotransmission and bipolar disorder: a systematic family-based association study.
Neurotransmission pathways/systems have been proposed to be involved in the pathophysiology and treatment of bipolar disorder for over 40 years. In order to test the hypothesis that common variants of genes in one or more of five neurotransmission systems confer risk for bipolar disorder, we analyzed 1,005 tag single nucleotide polymorphisms in 90 genes from dopaminergic, serotonergic, noradrenergic, GABAergic, and glutamatergic neurotransmitter systems in 101 trios and 203 quads from Caucasian bipolar families. Our sample has 80% power to detect ORs >or= 1.82 and >or=1.57 for minor allele frequencies of 0.1 and 0.5, respectively. Nominally significant allelic and haplotypic associations were found for genes from each neurotransmission system, with several reaching gene-wide significance (allelic: GRIA1, GRIN2D, and QDPR; haplotypic: GRIN2C, QDPR, and SLC6A3). However, none of these associations survived correction for multiple testing in an individual system, or in all systems considered together. Significant single nucleotide polymorphism associations were not found with sub-phenotypes (alcoholism, psychosis, substance abuse, and suicide attempts) or significant gene-gene interactions. These results suggest that, within the detectable odds ratios of this study, common variants of the selected genes in the five neurotransmission systems do not play major roles in influencing the risk for bipolar disorder or comorbid sub-phenotypes.


An autophagy-driven pathway of ATP secretion supports the aggressive phenotype of BRAFV600E inhibitor-resistant metastatic melanoma cells.
The ingrained capacity of melanoma cells to rapidly evolve toward an aggressive phenotype is manifested by their increased ability to develop drug-resistance, evident in the case of vemurafenib, a therapeutic-agent targeting BRAFV600E. Previous studies indicated a tight correlation between heightened melanoma-associated macroautophagy/autophagy and acquired Vemurafenib resistance. However, how this vesicular trafficking pathway supports Vemurafenib resistance remains unclear. Here, using isogenic human and murine melanoma cell lines of Vemurafenib-resistant and patient-derived melanoma cells with primary resistance to the BRAFV600E inhibitor, we found that the enhanced migration and invasion of the resistant melanoma cells correlated with an enhanced autophagic capacity and autophagosome-mediated secretion of ATP. Extracellular ATP (eATP) was instrumental for the invasive phenotype and the expansion of a subset of Vemurafenib-resistant melanoma cells. Compromising the heightened autophagy in these BRAFV600E inhibitor-resistant melanoma cells through the knockdown of different autophagy genes (ATG5, ATG7, ULK1), reduced their invasive and eATP-secreting capacity. Furthermore, eATP promoted the aggressive nature of the BRAFV600E inhibitor-resistant melanoma cells by signaling through the purinergic receptor P2RX7. This autophagy-propelled eATP-dependent autocrine-paracrine pathway supported the maintenance and expansion of a drug-resistant melanoma phenotype. In conclusion, we have identified an autophagy-driven response that relies on the secretion of ATP to drive P2RX7-based migration and expansion of the Vemurafenib-resistant phenotype. This emphasizes the potential of targeting autophagy in the treatment and management of metastatic melanoma.


Drugs and firearm deaths in New York City, 1990-1998.
Firearm deaths remain among the leading causes of mortality in the United States. Changing law enforcement activities, incarceration, drug use, and socioeconomic conditions may have played roles in the declining rates of firearm deaths during the 1990s. Using records from the Office of the Chief Medical Examiner, we analyzed the role of drugs in firearm deaths in New York City between 1990 and 1998. Positive drug toxicology was present in over half of all firearm death victims during this time. Cocaine, cannabis, opiates, and alcohol accounted for almost all of these deaths with drug-positive toxicology. There were decreases in cocaine- and alcohol-positive toxicology for firearm deaths in New York City starting in the early 1990s; there was a more gradual decrease in heroin-positive toxicology for firearm deaths. Cannabis-positive toxicology for firearm deaths increased in the early part of the 1990s and then decreased starting in the mid-1990s. Although the disparities between minority and white firearm death rates narrowed during this time, minorities remained about three times more likely to be victims of fatal firearm violence than whites in 1998. The highest firearm death rates were among African American and Latino male decedents, with a larger proportion of Latinos testing cocaine or opiate positive, while a larger proportion of African Americans tested cannabis positive. These results suggest a complex role of drugs in firearm-related deaths.


Nerve growth factor is primarily produced by GABAergic neurons of the adult rat cortex.
Within the cortex, nerve growth factor (NGF) mediates the innervation of cholinergic neurons during development, maintains cholinergic corticopetal projections during adulthood and modulates cholinergic function through phenotypic control of the cholinergic gene locus. Recent studies suggest NGF may also play an important role in cortical plasticity in adulthood. Previously, NGF-producing cells have been shown to colocalize with GABAergic cell markers within the hippocampus, striatum, and basal forebrain. Classification of cells producing NGF in the cortex is lacking, however, and cholinergic corticopetal projections have been shown to innervate both pyramidal and GABAergic neurons in the cortex. In order to clarify potential trophic interactions between cortical neurons and cholinergic projections, we used double-fluorescent immunohistochemistry to classify NGF-expressing cells in several cortical regions, including the prefrontal cortex, primary motor cortex, parietal cortex and temporal cortex. Our results show that NGF colocalizes extensively with GABAergic cell markers in all cortical regions examined, with >91% of NGF-labeled cells coexpressing GAD65/67. Conversely, NGF-labeled cells exhibit very little co-localization with the excitatory cell marker CaMKIIα (<5% of cells expressing NGF). NGF expression was present in 56% of GAD-labeled cells, suggesting that production is confined to a specific subset of GABAergic neurons. These findings demonstrate that GABAergic cells are the primary source of NGF production in the cortex, and likely support the maintenance and function of basal forebrain cholinergic projections in adulthood.


Incidence of chronic obstructive pulmonary disease in people with HIV in Ontario, 1996-2015: a retrospective population-based cohort study.
Because of high smoking rates and HIV-related factors, people with HIV may be at high risk for chronic obstructive pulmonary disease (COPD); however, population-based estimates of the incidence of COPD among people with HIV are lacking, particularly for women. We compared the incidence of COPD among Ontario adults aged 35 years or more with and without HIV between Jan. 1, 1996, and Dec. 31, 2015. We conducted a population-based study using Ontario's health administrative databases. We compared the incidence of COPD between people with and without HIV using standardized incidence ratios and generalized estimating equations with a log link function. We identified 1849 people with HIV and 1 168 727 HIV-negative people who were newly diagnosed with COPD between 1996 and 2015. People with HIV were younger than HIV-negative people (mean age 49.7 [standard deviation 10.4] yr v. 62.2 [standard deviation 14.8] yr; standardized difference 0.98). Rates of COPD were higher among people with HIV than among HIV-negative people (10.4 v. 9.0 cases per 1000 person-years; standardized incidence ratio 1.16, 95% confidence interval [CI] 1.10 to 1.21; adjusted rate ratio 1.34, 95% CI 1.27 to 1.41). In sex-stratified analyses, rates of COPD were higher among men with HIV (adjusted rate ratio 1.32, 95% CI 1.24 to 1.40) and women with HIV (adjusted rate ratio 1.54, 95% CI 1.37 to 1.72) than among men and women without HIV. In a sensitivity analysis, smoking explained observed differences in COPD incidence. People with HIV had higher rates of incident COPD than HIV-negative people. This may reflect the disproportionately higher prevalence of smoking among the former.


Pub, a novel PU.1 binding protein, regulates the transcriptional activity of PU.1.
PU.1 is a member of the Ets family of transcription factors and plays critical roles in the development of hematopoietic cells such as macrophages and B cells. To elucidate the molecular mechanism(s) underlying the regulation of PU.1 function, we screened for PU.1 interacting proteins using a yeast two-hybrid approach. As a result, a novel PU.1 binding factor, which we termed Pub, was isolated. The Pub protein has one B-box zinc finger domain, followed by a coiled-coil region and a B30.2-like domain, these features being characteristic of the tripartite motif (TRIM) family of protein. The PEST domain of PU.1 was found to interact with the N-terminal portion of Pub, a region that includes the TRIM which is considered to mediate protein-protein interactions. Northern blot and RT-PCR analyses demonstrated that Pub is predominantly expressed in hematopoietic tissues and cells where PU.1 is also expressed. Using a luciferase-based assay, we showed that Pub inhibited the transcriptional activity of PU.1. Moreover, the B-box zinc finger domain of Pub was critical for this inhibitory activity. These data suggest that Pub may be important in regulating the transcriptional activity of PU.1.


Effect of the ethinylestradiol/norelgestromin contraceptive patch on body composition. Results of bioelectrical impedance analysis in a population of Italian women.
As weight gain is one of the most frequently cited reasons for not using and for discontinuing hormonal contraceptives, in an open-label, single-arm, multicentre clinical study we evaluated the effect of the ethinylestradiol/norelgestromin contraceptive patch (EVRA, Janssen-Cilag International, Belgium) on body composition using bioelectrical impedance analysis (BIA). Body weight and impedance vector components (resistance (R) and reactance (Xc), at 50 kHz frequency, Akern-RJL Systems analyzer) were recorded before entry, after 1, 3 and 6 months in 182 Italian healthy women aged 29 yr (18 to 45), and with BMI 21.8 kg/m2 (16 to 31). Total body water (TBW) was estimated with a BIA regression equation. Vector BIA was performed with the RXc mean graph method and the Hotelling's T2 test for paired and unpaired data. After 6 months body weight increased by 0.64 kg (1.1%) and TBW increased by 0.51 L (1.7%). The pattern of impedance vector displacement indicated a small increase in soft tissue hydration (interstitial gel fluid). Body composition changes did not significantly differ among groups of previous contraceptive methods. Arterial blood pressure did not significantly change over time. After 6 months of treatment with the ethinylestradiol/norelgestromin contraceptive patch we found a minimal, clinically not relevant, increase in body weight less than 1 kg that could be attributed to an adaptive interstitial gel hydration. This fluctuation is physiological as confirmed by the lack of any effect on blood pressure. This could be useful in increasing women's choice, acceptability and compliance of the ethinylestradiol/norelgestromin contraceptive patch.


Structure-antiemetic-activity of some diarylheptanoids and their analogues.
The structure-activity relationship of diarylheptanoids and their analogues inhibitory of emesis induced by copper sulfate in young chicks was investigated by testing 19 compounds. The compounds are 5 diarylheptanoids isolated from Alpinia katsumadai (Zingiberacea), 5 chemical derivatives of them, 6 analogues isolated from Zingiber officinale rhizome (Zingiberaceae), and 3 analogues available on the market. Among them, two types of essential functional structure of diarylheptanoids and their analogues showed the inhibitory effects against emesis.


Genes influencing variation in serum osteocalcin concentrations are linked to markers on chromosomes 16q and 20q.
Osteocalcin (OC) is an important constituent of bone that is synthesized by osteoblasts. Serum levels of OC have been used as a biochemical marker of bone turnover. To identify the genes influencing variation in serum OC levels, we conducted a genome-wide scan in 429 individuals comprising 10 large multigenerational families. OC levels were measured by immunoassay, and genetic markers were typed at approximately 10-cM intervals across the genome. Quantitative trait linkage was tested using a multipoint analysis based on variance component methodology, adjusting for the effects of age, sex, and oral contraceptive use. Significance levels for linkage were obtained empirically, by Monte Carlo simulation. The heritability of OC levels in this population was 62 +/- 8%. We detected significant evidence for linkage between a quantitative trait locus influencing serum OC levels and markers on chromosome 16q, and suggestive evidence for linkage of OC levels with markers on chromosome 20q. The multipoint lod scores peaked at 3.35 on chromosome 16 and 2.78 on chromosome 20, corresponding to P values of 0.00004 and 0.00017, respectively. A potential candidate gene for bone formation in the linked region on chromosome 20 is CDMP1, which encodes cartilage-derived morphogenetic protein 1. Future studies should evaluate whether variation in CDMP1 or in other genes in the linked regions on chromosomes 16 and 20 influence the rate of bone turnover.


GeneWeaver: finding consilience in heterogeneous cross-species functional genomics data.
A persistent challenge lies in the interpretation of consensus and discord from functional genomics experimentation. Harmonizing and analyzing this data will enable investigators to discover relations of many genes to many diseases, and from many phenotypes and experimental paradigms to many diseases through their genomic substrates. The GeneWeaver.org system provides a platform for cross-species integration and interrogation of heterogeneous curated and experimentally derived functional genomics data. GeneWeaver enables researchers to store, share, analyze, and compare results of their own genome-wide functional genomics experiments in an environment containing rich companion data obtained from major curated repositories, including the Mouse Genome Database and other model organism databases, along with derived data from highly specialized resources, publications, and user submissions. The data, largely consisting of gene sets and putative biological networks, are mapped onto one another through gene identifiers and homology across species. A versatile suite of interactive tools enables investigators to perform a variety of set analysis operations to find consilience among these often noisy experimental results. Fast algorithms enable real-time analysis of large queries. Specific applications include prioritizing candidate genes for quantitative trait loci, identifying biologically valid mouse models and phenotypic assays for human disease, finding the common biological substrates of related diseases, classifying experiments and the biological concepts they represent from empirical data, and applying patterns of genomic evidence to implicate novel genes in disease. These results illustrate an alternative to strict emphasis on replicability, whereby researchers classify experimental results to identify the conditions that lead to their similarity.


Modulation of uPA, MMPs and their inhibitors by a novel nutrient mixture in human glioblastoma cell lines.
Brain tumors are highly aggressive tumors that are characterized by high levels of matrix metalloproteinase (MMP)-2 and -9 secretions that degrade the extracellular matrix (ECM) and basement membrane, allowing cancer cells to spread to distal organs. Proteases play a key role in tumor cell invasion and metastasis by digesting the basement membrane and ECM components. Strong clinical and experimental evidence demonstrates association of elevated levels of urokinase plasminogen activators (uPA) and MMPs with cancer progression, metastasis and shortened patient survival. MMP activities are regulated by specific tissue inhibitors of metalloproteinases (TIMPs). Our main objective was to study the effect of a nutrient mixture (NM) on the activity of uPA, MMPs and TIMPs in various human gliomas. Human glioblastoma (LN-18, T-98G and A-172) cell lines (ATCC) were cultured in their respective media and treated at confluence with NM at 0, 50, 100, 250, 500 and 1000 µg/ml. Analysis of uPA activity was carried out by fibrin zymography, MMPs by gelatinase zymography and TIMPs by reverse zymography. Glioblastoma cell lines LN-18 and T-98G expressed uPA, which was inhibited by NM in a dose-dependent manner. However, no bands corresponding to uPA were detected for the A-172 cell line. On gelatinase zymography, all three cell lines showed bands corresponding to MMP-2 and LN-18 and T-98G showed PMA (100 ng/ml)-induced MMP-9. NM inhibited their expression in a dose-dependent manner. Activity of TIMP-2 was upregulated by NM in all glioma cell lines in a dose-dependent manner. Analysis revealed a positive correlation between uPA and MMP-2 and a negative correlation between uPA/MMPs and TIMP-2. These findings suggest the therapeutic potential of NM in the treatment of gliomas.


Function of Rho GTPases in embryonic blood cell migration in Drosophila.
Hemocyte development in the Drosophila embryo is a genetic model to study blood cell differentiation, cell migration and phagocytosis. Macrophages, which make up the majority of embryonic hemocytes, migrate extensively as individual cells on basement membrane-covered surfaces. The molecular mechanisms that contribute to this migration process are currently not well understood. We report the generation, by P element replacement, of two Gal4 lines that drive expression of UAS-controlled target genes during early (gcm-Gal4) or late (Coll-Gal4) stages of macrophage migration. gcm-Gal4 is used for live imaging analysis showing that macrophages extend large, dynamic lamellipodia as their main protrusions as well as filopodia. We use both Gal4 lines to express dominantnegative and constitutively active isoforms of the Rho GTPases Rac1, Cdc42, Rho1 and RhoL in macrophages, and complement these experiments by analyzing embryos mutant for Rho GTPases. Our findings suggest that Rac1 and Rac2 act redundantly in controlling migration and lamellipodia formation in Drosophila macrophages, and that the third Drosophila Rac gene, Mtl, makes no significant contribution to macrophage migration. Cdc42 appears not to be required within macrophages but in other tissues of the embryo to guide macrophages to the ventral trunk region. No evidence was found for a requirement of Rho1 or RhoL in macrophage migration. Finally, to estimate the number of genes whose zygotic expression is required for macrophage migration we analyzed 208 chromosomal deletions that cover most of the Drosophila genome. We find eight deletions that cause defects in macrophage migration suggesting the existence of approximately ten zygotic genes essential for macrophage migration.


Identification of the regions of PECAM-1 involved in beta- and gamma-catenin associations.
Platelet endothelial cell adhesion molecule-1 (PECAM-1) binds tyrosine-phosphorylated beta-catenin and modulates beta-catenin localization and sequestration. The biological significance of this interaction, while still unclear, it has been postulated to be involved in modulating adherens junction dynamics in response to perturbants [J. Clin. Invest. 109 (2002) 383]. Here we demonstrate that tyrosine-phosphorylated beta-catenin, and to a lesser extent unphosphorylated beta-catenin, interact with a portion of the cytoplasmic domain of PECAM-1 encoded by exon 15. Using RT-PCR, we obtained products representing alternatively spliced PECAM-1 isoforms from mouse kidney total mRNA and generated PECAM-1-GST constructs expressing full length and naturally occurring alternatively spliced PECAM-1 variants. Co-precipitation assays revealed that the protein sequence encoded by exon 15 is necessary for beta-catenin binding. Transfections using deletion mutants confirmed the importance of the exon 15 sequence in this interaction. In contrast, gamma-catenin-PECAM-1 interactions are thought to be modulated by an as yet undefined PECAM-1 serine phosphorylation and appear to mediate dynamic PECAM-1 intermediate filament cytoskeletal interactions [J. Biol. Chem. 275 (2000) 21435]. Here we demonstrate that the PECAM-1-gamma-catenin interaction occurs via an exon 13-mediated process. GST-pull-down assays illustrated the importance of the exon 13 sequence in this interaction. Further, using site-directed mutagenesis of S(673) to C and D and S(669 and 670) to C, we confirmed the importance of S(673) and its phosphorylation state as a mediator of gamma-catenin-PECAM-1 binding. Our studies define the exons of the PECAM-1 cytoplasmic domain that is involved in mediating these PECAM-1-catenin family member interactions and will allow investigators to better define the biological functions resulting from these interactions.


Proventil HFA and ventolin have similar safety profiles during regular use.
As a secondary objective to a long-term study evaluating the bronchodilator effectiveness of Proventil HFA (albuterol), to assess the safety of Proventil HFA, Ventolin, and hydrofluoroalkane 134a (HFA-134a) placebo over 12 weeks of regular dosing. Randomized, double-blind, double-dummy parallel group, placebo-controlled, multicenter trial of asthmatics requiring inhaled beta-adrenergic bronchodilators for symptom control. Treatment with Proventil HFA, Ventolin, or HFA-134a placebo, qid, for 12 weeks. Adverse events were reviewed at biweekly clinic visits. Between clinic visits, patients recorded morning and evening peak expiratory flow (PEF), asthma symptom and nighttime asthma sleep disturbance scores, and use of rescue beta-adrenergic bronchodilator on diary cards daily. Investigators provided a global assessment of asthma control at weeks 0, 4, 8, and 12. Vital signs were recorded over 6 h after dosing with study drug at weeks 0, 4, 8, and 12. Standard laboratory tests, CBC count, serum chemistries, and urinalysis were obtained at study start and end. Adverse event reporting rates were similar for the three treatment groups. The morning PEF tended to be lower for the Proventil HFA and Ventolin groups than the HFA-134a placebo group, but the evening PEF tended to be higher for the active treatment groups. Daytime asthma symptom scores tended to be lower (better) with active treatment than placebo, but nighttime asthma sleep disturbance scores were similar for all three treatment groups. Use of Ventolin Rotacaps as rescue medication was significantly greater for the HFA-134a placebo group than the Proventil HFA and Ventolin groups. Diary card data did not change within groups over time. Investigator global assessments of asthma scores clustered between fair and good for all three treatment groups throughout the study. Changes in heart rate and BP were small after dosing with study drug and tended to be similar for the active treatments and HFA-134a placebo groups. No clinically meaningful changes in results of standard laboratory tests were found in any treatment group during this study. Proventil HFA had a similar safety profile as Ventolin during regular use. A dosage of 16 puffs per day of propellant HFA-134a was well tolerated by asthmatics. Regular use of either Proventil HFA or Ventolin did not cause asthma control to deteriorate.


Neurons containing gastrin releasing peptide-like immunoreactivity in the human pancreas.
Gastrin releasing peptide (GRP) is known to stimulate pancreatic enzyme and islet hormone secretion. In the present immunohistochemical study, the localization and distribution of GRP-like immunoreactivity were investigated in the human pancreas using two antisera with different specificities. GRP-like immunoreactivity (GRP-LI) was observed in numerous nerve fibers diffusely distributed to the exocrine pancreas, but was not seen in intrapancreatic nerve cells of normal pancreatic specimens examined. Nerve fibers and terminals with GRP-LI were found in abundance around pancreatic acini and capillaries, with moderate density around ductules and in the walls of arterioles, and a few were seen in islets. This distribution pattern was quite similar to that of vasoactive intestinal polypeptide (VIP)-LI nerve fibers. The study, using the antibody elution method, strongly suggests the co-localization of GRP- and VIP-LIs within a part of VIP-containing nerve fibers. In the chronic pancreatitis specimens, neurons with GRP-LI were frequently found, and > 90% of intrapancreatic nerve cells were VIP-immunoreactive. Immunostainings for GRP and for VIP on serial adjacent sections of intrapancreatic ganglia from chronic pancreatitis specimens suggested the co-localization of the two immunoreactivities in > 70% of intrapancreatic neurons. The present findings may provide a morphological basis for neurotransmitter and/or neuromodulator roles of GRP in the human pancreas.


Biphasic response of a tecto-mesencephalic pilocytic astrocytoma after Gamma Knife surgery--A case report.
Biphasic response (shrinkage-regrowth-shrinkage) of tumors has never previously been reported in the postoperative course, neither after microsurgery, nor after Gamma Knife surgery (GKS). We present the case of an adult with dorsal midbrain syndrome resulting from a pilocytic astrocytoma centered on the mesencephalic tectum. The tumor extended to the third ventricle and the thalamus. Initially, due to tumor growth, a biopsy was performed and histology established. Later, a ventriculocisternostomy for obstructive hydrocephalus was performed. Finally, GKS was performed, as the tumor continued to grow. After GKS, the lesion exhibited a biphasic response, with a major shrinkage at 3 months, regrowth within the target volume at 6 and 9 months and a second phase of important shrinkage at 12 months, which persisted for the next two years. The possible mechanisms for this particular response pattern are discussed.


Two-electrode gas switch with electrodynamical acceleration of a discharge channel.
High-energy switches and trigger generators are required for MJ-level capacitor banks. We have developed a compact gas switch and a matched series injection trigger generator. A series inductance is required for isolation of the trigger pulse from the surrounded circuit. A saturable inductor is employed here because low inductance is needed after the switch breakdown. The switch is of coaxial two-electrode design with electrodynamic acceleration of a spark channel. The switch operates at atmospheric pressure. The spark gap can be triggered reliably down to zero voltage (at 50 kV self-breakdown voltage) with less than 35 ns timing jitter. Energy losses in this spark gap have been accurately investigated. The main results are as follows: energy losses in the switch do not exceed 4% at voltages higher than 15 kV, i.e., when operation voltage exceeds ∼36% of the self-breakdown voltage. The spark gap is designed for 24 kV charging voltage, at a current up to 250 kA, and ∼70 C charge transfer. In this paper, we present a design for the spark gap, inductor and trigger generator. Test bed schematics and results of the tests are also described.


Oncogenic GNAQ and GNA11 mutations in uveal melanoma in Chinese.
To examine whether GNAQ and GNA11 somatic mutations previously identified in uveal melanomas of Caucasians are associated with uveal melanomas in Chinese patients. Uveal melanomas treated by primary enucleation in Chinese patients underwent a mutation analysis of GNAQ and GNA11 with sequencing of exon 5 and exon 4. The study included 50 patients with uveal melanoma and with a mean age of 47.6±13.0 years. During the follow-up of at least 3 years, 20 (40%) patients developed extraocular metastases. The frequencies of GNAQ and GNA11 somatic mutations in uveal melanoma were 18% (9/50) and 20% (10/50), respectively. The mutations occurred exclusively in codon 209 of exon 5. No mutations were detected in exon 4. Mutations affecting codon 209 in GNAQ were c.626A>C(Q209P) (78%) and c.626A>T(Q209L) (22%). Mutations affecting codon 209 in GNA11 were exclusively c.626A>T(Q209L) (100%). In none of the tumors, mutations of BRAF and NRAS were detected. GNAQ/11 mutations were marginally (P = 0.045) associated with optic disc involvement. In Kaplan-Meier analysis, metastasis-free survival was not significantly (P = 0.94) associated with GNAQ/11 mutations. Mutations of GNAQ and GNA11 can be found in Chinese patients as in Caucasian patients with uveal melanoma, with a higher frequency reported for Caucasian patients.


Eph receptor signalling: from catalytic to non-catalytic functions.
Eph receptors, the largest subfamily of receptor tyrosine kinases, are linked with proliferative disease, such as cancer, as a result of their deregulated expression or mutation. Unlike other tyrosine kinases that have been clinically targeted, the development of therapeutics against Eph receptors remains at a relatively early stage. The major reason is the limited understanding on the Eph receptor regulatory mechanisms at a molecular level. The complexity in understanding Eph signalling in cells arises due to following reasons: (1) Eph receptors comprise 14 members, two of which are pseudokinases, EphA10 and EphB6, with relatively uncharacterised function; (2) activation of Eph receptors results in dimerisation, oligomerisation and formation of clustered signalling centres at the plasma membrane, which can comprise different combinations of Eph receptors, leading to diverse downstream signalling outputs; (3) the non-catalytic functions of Eph receptors have been overlooked. This review provides a structural perspective of the intricate molecular mechanisms that drive Eph receptor signalling, and investigates the contribution of intra- and inter-molecular interactions between Eph receptors intracellular domains and their major binding partners. We focus on the non-catalytic functions of Eph receptors with relevance to cancer, which are further substantiated by exploring the role of the two pseudokinase Eph receptors, EphA10 and EphB6. Throughout this review, we carefully analyse and reconcile the existing/conflicting data in the field, to allow researchers to further the current understanding of Eph receptor signalling.


Structural conservation of neurotropism-associated VspA within the variable Borrelia Vsp-OspC lipoprotein family.
Vsp surface lipoproteins are serotype-defining antigens of relapsing fever spirochetes that undergo multiphasic antigenic variation to avoid the immune response. One of these proteins, VspA of Borrelia turicatae, is also associated with neurotropism in infected mice. Vsp proteins are highly polymorphic in sequence, which may relate to their specific antibody reactivities and host cell interactions. To determine whether sequence variations affect protein structure, we compared B. turicatae VspA with three related proteins: VspB of B. turicatae, Vsp26 of the relapsing fever agent Borrelia hermsii, and OspC of the Lyme disease spirochete Borrelia burgdorferi. Recombinant non-lipidated proteins were purified by affinity or ion exchange chromatography. Circular dichroism spectra revealed similar, highly alpha-helical secondary structures for all four proteins. In vitro assays demonstrated protease-resistant, thermostable Vsp cores starting at a conserved serine at position 34 (Ser(34)). All proteins aggregate as dimers in solution. In situ trypsin treatment and surface protein cross-linking showed that the native lipoproteins also form protease-resistant dimers. These findings indicate that Vsp proteins have a common compact fold and that their established functions are based on localized polymorphisms. Two forms of VspA crystals suitable for structure determination by x-ray diffraction methods have been obtained.


Collaborative mental health in rural and isolated Canada: stakeholder feedback.
This article presents research findings from the Rural and Isolated Working Group, one of six groups established by the Canadian Collaborative Mental Health Initiative (CCMHI). Funded through Health Canada's Primary Health Care Transition Fund, the goal of the CCMHI is to improve the mental health and well-being of Canadians by increasing collaboration among primary health care and mental health care providers, consumers, families, and caregivers. Qualitative data obtained from mental health care providers and consumers across all regions of Canada are presented in this article. Policy and regulation problems, barriers to mental health care access, service providers' perspectives of the challenges to consumer involvement, and solutions for addressing these issues are discussed. The article concludes by identifying how this research has informed and influenced initial steps toward mental health promotion and treatment of mental illness in rural and isolated Canada.


Changes in muscle activation following balance and technique training and a season of Australian football.
Determine if balance and technique training implemented adjunct to 1001 male Australian football players' training influenced the activation/strength of the muscles crossing the knee during pre-planned and unplanned sidestepping. Randomized Control Trial. Each Australian football player participated in either 28 weeks of balance and technique training or 'sham' training. Twenty-eight Australian football players (balance and technique training, n=12; 'sham' training, n=16) completed biomechanical testing pre-to-post training. Peak knee moments and directed co-contraction ratios in three degrees of freedom, as well as total muscle activation were calculated during pre-planned and unplanned sidestepping. No significant differences in muscle activation/strength were observed between the 'sham' training and balance and technique training groups. Following a season of Australian football, knee extensor (p=0.023) and semimembranosus (p=0.006) muscle activation increased during both pre-planned sidestepping and unplanned sidestepping. Following a season of Australian football, total muscle activation was 30% lower and peak valgus knee moments 80% greater (p=0.022) during unplanned sidestepping when compared with pre-planned sidestepping. When implemented in a community level training environment, balance and technique training was not effective in changing the activation of the muscles crossing the knee during sidestepping. Following a season of Australian football, players are better able to support both frontal and sagittal plane knee moments. When compared to pre-planned sidestepping, Australian football players may be at increased risk of anterior cruciate ligament injury during unplanned sidestepping in the latter half of an Australian football season.


Oocyte Cryopreservation for Fertility Preservation in Healthy Women.
Advancements in methods of fertility preservation have resulted in more options available to women seeking to postpone reproduction. Preservation of unfertilized oocytes provides women with the possibility for reproduction even after age-related declines in fertility might make conceiving a child difficult to impossible. As oocyte cryopreservation rises in popularity among women, it is crucial for nurses and other clinicians to understand the background, process, ethical issues, and risks involved to help women make informed medical decisions.


Spatial diffusivity and availability of intracellular calmodulin.
Calmodulin (CaM) is the major pathway that transduces intracellular Ca2+ increases to the activation of a wide variety of downstream signaling enzymes. CaM and its target proteins form an integrated signaling network believed to be tuned spatially and temporally to control CaM's ability to appropriately pass signaling events downstream. Here, we report the spatial diffusivity and availability of CaM labeled with enhanced green fluorescent protein (eGFP)-CaM, at basal and elevated Ca2+,quantified by the novel fluorescent techniques of raster image scanning spectroscopy and number and brightness analysis. Our results show that in basal Ca2+ conditions cytoplasmic eGFP-CaM diffuses at a rate of 10 microm(2)/s, twofold slower than the noninteracting tracer, eGFP, indicating that a significant fraction of CaM is diffusing bound to other partners. The diffusion rate of eGFP-CaM is reduced to 7 microm(2)/s when a large (646 kDa) target protein Ca2+/CaM-dependent protein kinase II is coexpressed in the cells. In addition, the presence of Ca2+/calmodulin-dependent protein kinase II, which can bind up to 12 CaM molecules per holoenzyme, increases the stoichiometry of binding to an average of 3 CaMs per diffusive molecule. Elevating intracellular Ca2+ did not have a major impact on the diffusion of CaM complexes. These results present us with a model whereby CaM is spatially modulated by target proteins and support the hypothesis that CaM availability is a limiting factor in the network of CaM-signaling enzymes.


Genome-wide survey of codons under diversifying selection in a highly recombining bacterial species, Helicobacter pylori.
Selection has been a central issue in biology in eukaryotes as well as prokaryotes. Inference of selection in recombining bacterial species, compared with clonal ones, has been a challenge. It is not known how codons under diversifying selection are distributed along the chromosome or among functional categories or how frequently such codons are subject to mutual homologous recombination. Here, we explored these questions by analysing genes present in >90% among 29 genomes of Helicobacter pylori, one of the bacterial species with the highest mutation and recombination rates. By a method for recombining sequences, we identified codons under diversifying selection (dN/dS> 1), which were widely distributed and accounted for ∼0.2% of all the codons of the genome. The codons were enriched in genes of host interaction/cell surface and genome maintenance (DNA replication,recombination, repair, and restriction modification system). The encoded amino acid residues were sometimes found adjacent to critical catalytic/binding residues in protein structures.Furthermore, by estimating the intensity of homologous recombination at a single nucleotide level, we found that these codons appear to be more frequently subject to recombination.We expect that the present study provides a new approach to population genomics of selection in recombining prokaryotes.


Acetylcholine functionally reorganizes neocortical microcircuits.
Sensory information is processed and transmitted through the synaptic structure of local cortical circuits, but it is unclear how modulation of this architecture influences the cortical representation of sensory stimuli. Acetylcholine (ACh) promotes attention and arousal and is thought to increase the signal-to-noise ratio of sensory input in primary sensory cortices. Using high-speed two-photon calcium imaging in a thalamocortical somatosensory slice preparation, we recorded action potential activity of up to 900 neurons simultaneously and compared local cortical circuit activations with and without bath presence of ACh. We found that ACh reduced weak pairwise relationships and excluded neurons that were already unreliable during circuit activity. Using action potential activity from the imaged population, we generated functional wiring diagrams based on the statistical dependencies of activity between neurons. ACh pruned weak functional connections from spontaneous circuit activations and yielded a more modular and hierarchical circuit structure, which biased activity to flow in a more feedforward fashion. Neurons that were active in response to thalamic input had reduced pairwise dependencies overall, but strong correlations were conserved. This coincided with a prolonged period during which neurons showed temporally precise responses to thalamic input. Our results demonstrate that ACh reorganizes functional circuit structure in a manner that may enhance the integration and discriminability of thalamic afferent input within local neocortical circuitry.


Multicenter analysis of impact of anastomotic leakage on long-term oncologic outcomes after curative resection of colon cancer.
In rectal cancer, anastomotic leakage was reported to have a negative impact on both short- and long-term outcomes. However, there is limited data on the impact of anastomotic leakage on oncologic outcomes in patients with colon cancer. We aimed to evaluate the impact of anastomotic leakage on disease recurrence and long-term survival after curative resection of colon cancer. This multicenter, retrospective cohort study of 4,919 consecutive patients utilized data from the Japanese Society for Cancer of the Colon and Rectum. Multivariable Cox regression analysis was used to adjust for confounding. The incidence of anastomotic leakage was 2.5% and 30-day mortality was 0.21%. The 5-year overall survival rate was 80.8% in the anastomotic leakage group, compared with 90.3% in the no leak group (P = .001). In the multivariable analysis, anastomotic leakage was significantly associated with reduced overall survival rate (hazard ratio = 1.84; 95% confidence interval, 1.06-2.96). Overall disease recurrence rate was 14.1%: 21.2% in the anastomotic leakage group and 13.9% in the no leak group. There was a significant association between anastomotic leakage and local recurrence (hazard ratio = 4.63; 95% confidence interval, 1.60-10.6). In contrast, anastomotic leakage was not significantly associated with total distant recurrence. However, anastomotic leakage did show a tendency toward increasing peritoneal recurrence, although it did not reach statistical significance (hazard ratio = 2.59; 95% confidence interval, 0.79-6.29). In our study population, anastomotic leakage was associated with reduced overall survival and with increased rate of local recurrence after curative resection for colon cancer.


Increased Risk of Traffic Injury After a Cerebrovascular Event.
Background and Purpose- We aimed to determine the long-term risks of a motor vehicle collision after a cerebrovascular event and whether the risks were similar after left- or right-hemispheric events. Methods- We used a population-based registry to identify patients diagnosed with a transient ischemic attack or stroke (hemorrhagic or ischemic) between 2003 and 2013 in Ontario, Canada. Hemispheric laterality was determined using radiological and clinical findings. We identified subsequent serious injuries involving the patient as a driver using linked administrative data. Secondary outcomes included serious injuries involving the patient as a pedestrian, as a passenger, or other traumatic events (fall, fracture, ankle sprain). We used proportional hazard models accounting for death as a competing risk to test the association of hemispheric laterality and outcomes with and without adjustment for age, sex, discharge modified Rankin Scale score, home location, and prior driving record. Patients were followed through to 2017. Results- Among 26 144 patients with hemispheric cerebrovascular events, 377 subsequent serious traffic injuries as a driver (2.2 per 1000 person-year) were identified over a median follow-up of 6.4 person-years. The rate did not differ by laterality (adjusted hazard ratio, 1.00; 95% CI, 0.82-1.23). The risk of a serious traffic injury as a pedestrian was significantly higher after a right-sided than left-sided event (adjusted hazard ratio, 1.27; 95% CI, 1.02-1.58). Subsequent risks for other traumatic injuries did not differ by laterality of cerebrovascular event. Conclusions- The risk of a serious traffic injury as a pedestrian is substantially higher after a right-hemispheric cerebrovascular event compared with a left-sided event. Walking should be promoted for exercise in survivors of a stroke or transient ischemic attack, but these vulnerable road users may benefit from additional poststroke rehabilitation to optimize safety.


[Emotional stress as a clinical model to study the pathogenesis of the initial phase of the general adaptation syndrome].
General adaptation syndrome (GAS), the basis of the development of which is stress phenomenon, is an essential component of the pathogenesis of many diseases and syndromes. However, the patho genesis of GAS hitherto is considered exclusively from the endocrinological viewpoint. This relates primarily to the initial phase of the GAS, a clinical model for the study of which may be psycho-emotional stress (PES), which we studied using three groups of volunteers. The first one consists of 25 students who were waiting for unaccustomed physical activity (17 men) and play debut on the stage (8 women). The second group consists of 48 children (2-14 years) who expected for "planned" surgery. The third group of volunteers is made up of 80 students (41 women and 39 men) during the first exam. The concentration of cortisol, endotoxin (ET), the activity of antiendotoxin immunity (AEI) and the haemostatic system parameters were determined in the blood serum of volunteers in various combinations. We found laboratory evidence for PES at 92% of students of the first group, 58% of children of the second one and in 21% of students of the third group of volunteers (mostly women). The concentration of ET increased at 13 (52%) volunteers of the first group with a significant increase of average indicators in the whole group (from 0.84 ± 0.06 to 1.19 ± 0.04 EU/ml). At children of the second group, the average concentration of ET increased even more significantly (from 0.42 ± 0.02 to 1.63 ± 0.11 EU/ml), which was accompanied by the activation of the hemostasis system. A degree of the activation was directly dependent on the level of ET in the general circulation and on an activity of AEI. Examination stress in the third group of volunteers is accompanied by activation of plasma hemostasis (increased initial thrombosis rate and reduced the time it starts, lag-period) in 26% of female students and 15% of male students. We suggest that it is possible to use the PES as a clinical model for studying the initial phase of the GAS, examine the role of excess of intestinal ET in the general blood circulation (endotoxin aggression) in the induction of systemic inflammation, which is very likely participated in the initiation of the GAS.


Rearing strategy and optimizing first-calving targets in dairy heifers: a review.
Much research has been carried out and published on dairy replacement management, in order to rear heifers as efficiently as possible, from both a technical and economical point of view. In most cases, the aim is to rear the heifers at the lowest cost possible without any deleterious effects on future performances. However, the importance of dairy heifer husbandry is not sufficiently well recognized and probably mishandled by most farmers. The present review aims to give an actual overview of rearing procedures in dairy heifers and possible ways to achieve optimal goals. For many years, it has been well known that rapid rearing lowers the age of sexual maturity and consequently may be an efficient way to reduce the non-producing period prior to conception. But this may impair mammary development and consequently future milk production, at least during first lactation. In addition, a growth rate that is too low may not only be costly but also result in animals that are too fat at first calving, creating problems such as calving difficulties, dystocia, etc. Genetic considerations must also be factored, i.e. frame, size, body weight, etc. have changed during the last 20 years and there are differences between breeds. As a result, some time-honoured recommendations may not be appropriate. The present paper reviews factors and management practices that may affect rearing and subsequent performance of dairy heifers.


PET respiratory motion correction: quo vadis?
Positron emission tomography (PET) respiratory motion correction has been a subject of great interest for the last twenty years, prompted mainly by the development of multimodality imaging devices such as PET/computed tomography (CT) and PET/magnetic resonance imaging (MRI). PET respiratory motion correction involves a number of steps including acquisition synchronization, motion estimation and finally motion correction. The synchronization steps include the use of different external device systems or data driven approaches which have been gaining ground over the last few years. Patient specific or generic motion models using the respiratory synchronized datasets can be subsequently derived and used for correction either in the image space or within the image reconstruction process. Similar overall approaches can be considered and have been proposed for both PET/CT and PET/MRI devices. Certain variations in the case of PET/MRI include the use of MRI specific sequences for the registration of respiratory motion information. The proposed review includes a comprehensive coverage of all these areas of development in field of PET respiratory motion for different multimodality imaging devices and approaches in terms of synchronization, estimation and subsequent motion correction. Finally, a section on perspectives including the potential clinical usage of these approaches is included.


Pituitary gland agouti-related peptide cells: Novel player controlling glucose metabolism.
The liver synthesizes and releases bile acids into the gut. Bile acids, either directly or indirectly, inhibit agouti-related peptide (AGRP)-B cells in the pars tuberalis of the pituitary gland. AGRP-B cells are assumed to promote pancreatic insulin secretion and/or to improve insulin sensitivities in insulin sensitive organs, resulting in improved glucose tolerance.


The functional role of histidine and sulfhydryl groups in rat liver thiamine pyrophosphokinase.
The functional role of histidine and sulfhydryl groups of thiamine pyrophosphokinase (ATP: thiamine pyrophosphotransferase, E.C.220.127.116.11) has been studied by the methods of photooxidation and chemical modification by diethylpyrocarbonate (DEPC) and Ellman's reagent, 5,5' - dithiobis (2 - nitrobenzoic acid) (DTNB). Histidine amino acid residues have been shown to be destroyed during photoinactivation. Titration of the protein with DEPC has established that modification of two histidine groups decreases the catalytic activity of thiamine pyrophosphokinase. Interaction of the subsequent three groups with the reagent does not affect residual activity. Substrates protect thiamine pyrophosphokinase from inactivation. Two -SH groups have been identified in a molecule of thiamine pyrophosphokinase with Ellman's reagent. Modification of only one of them results in complete loss of the enzymatic activity. Treatment of the enzyme with 8M urea has shown no differences in the amount of thiol groups of the native and denatured enzymes. Mg2+ + ATP and somewhat more weakly Mg2+ partially protect thiamine pyrophosphokinase from inhibition by DTNB. In their presence a high degree of enzyme reactivation is observed. Both histidine and sulfhydryl groups are suggested to play an important role in the catalytic mechanism of thiamine pyrophosphokinase.


[Treatment of metastatic malignant melanoma localized to an extremity].
At the Norwegian Radium Hospital, most patients who were treated for metastases (localized to an extremity) from malignant melanoma in the period 1977-99 underwent intra-arterial chemotherapy combined with radiotherapy. We have evaluated the effects of this treatment, which has now been replaced by isolated limb perfusion (with melphalan and tumor necrosis factor). Medical records were reviewed for patients with metastatic malignant melanoma (localized to an extremity) who had been treated at the Norwegian Radium Hospital with intra-arterial chemotherapy (5-[3,3 dimethyl-1-triazeno]-imidazole-4-carboxamide [DTIC]) in the period 1977-99. 36 patients had received such treatment; in 30 of these the induction treatment was combined with radiation of the tumour area.. 24 patients were in complete remission after treatment (12 of these had all the metastases surgically removed before treatment). Nine patients had a partial remission while three patients had progressive disease. Median observation time was 49 months. 15 of 31 patients had an observation time of more than five years, and seven of these patients were alive after ten years. Three patients with metastasis localized to the lower extremity died of other causes. In one case the intra-arterial catheterisation caused serious aortic damage. It is possible to achieve long-term remission in patients with metastatic malignant melanoma localized to an extremity after intra-arterial chemotherapy combined with radiotherapy.


White matter organization in relation to upper limb motor control in healthy subjects: exploring the added value of diffusion kurtosis imaging.
Diffusion tensor imaging (DTI) characterizes white matter (WM) microstructure. In many brain regions, however, the assumption that the diffusion probability distribution is Gaussian may be invalid, even at low b values. Recently, diffusion kurtosis imaging (DKI) was suggested to more accurately estimate this distribution. We explored the added value of DKI in studying the relation between WM microstructure and upper limb coordination in healthy controls (N = 24). Performance on a complex bimanual tracking task was studied with respect to the conventional DTI measures (DKI or DTI derived) and kurtosis metrics of WM tracts/regions carrying efferent (motor) output from the brain, corpus callosum (CC) substructures and whole brain WM. For both estimation models, motor performance was associated with fractional anisotropy (FA) of the CC-genu, CC-body, the anterior limb of the internal capsule, and whole brain WM (r s range 0.42-0.63). Although DKI revealed higher mean, radial and axial diffusivity and lower FA than DTI (p < 0.001), the correlation coefficients were comparable. Finally, better motor performance was associated with increased mean and radial kurtosis and kurtosis anisotropy (r s range 0.43-0.55). In conclusion, DKI provided additional information, but did not show increased sensitivity to detect relations between WM microstructure and bimanual performance in healthy controls.


The influence of blinding, olfactory bulbectomy, and pinealectomy on twenty four-hour plasma prolactin levels in normal and neonatally androgenized female rats.
The influence of blinding (BLD), olfactory bulbectomy (ANOS), and pinealectomy (PX) on 24-h plasma PRL levels was examined in normal and neonatally androgenized (NA) female rats. NA pups were treated at 3 days of age with 1.25 mg testosterone propionate. Surgery was performed on both NA and normal animals at 22-25 days of age. At 10 weeks of age, animals were vaginally smeared, and at 15 weeks, they were ovariectomized, injected with 0.5 mg polyestradiol phosphate, and fitted with atrial catheters for blood sampling. At 16 weeks of age, blood samples were taken over a 24-h period. In normal animals, BLD plus ANOS resulted in half of the animals exhibiting a diestrous vaginal smear, while the other half exhibited at least one vaginal estrus during the 7-day period when vaginal smears were obtained. Ovarian, oviductal, and uterine weights for blinded (BLD) plus olfactory bulbectomized (ANOS) animals exhibiting diestrous smears were significantly less than those of BLD plus ANOS animals exhibiting periodic vaginal estrus. Plasma PRL levels were lower in both animal groups compared with those in sham controls, but BLD plus ANOS animals with small ovaries had significantly lower plasma PRL and BLD plus ANOS animals with large ovaries. PX of BLD plus ANOS animals resulted in endocrine organ weights comparable to those in controls and elevated the plasma PRL levels to those observed in BLD plus ANOS animals with large ovaries. The PRL levels in BLD, ANOS, and pinealectomized animals, however, were still below those in sham control animals. Sensory deprived animals had peak PRL values at various times of day, and regrouping of the data from the peak PRL time for each animal suggested the presence of a free-running plasma PRL rhythm. In NA animals, a decrease was also observed in endocrine organ weights and plasma PRL for BLD plus ANOS animals similar to that observed for normal BLD plus ANOS animals. However, all animals in the NA BLD plus ANOS group responded with a constant diestrous smear, small ovaries, and low plasma PRL values. PX prevented the decrease in endocrine organ weights and plasma PRL observed in BLD plus ANOS animals. As in normal animals, NA animals also exhibited a free-running PRL rhythm when sensory deprived.(ABSTRACT TRUNCATED AT 400 WORDS)


Biological control and sustainable food production.
The use of biological control for the management of pest insects pre-dates the modern pesticide era. The first major successes in biological control occurred with exotic pests controlled by natural enemy species collected from the country or area of origin of the pest (classical control). Augmentative control has been successfully applied against a range of open-field and greenhouse pests, and conservation biological control schemes have been developed with indigenous predators and parasitoids. The cost-benefit ratio for classical biological control is highly favourable (1:250) and for augmentative control is similar to that of insecticides (1:2-1:5), with much lower development costs. Over the past 120 years, more than 5000 introductions of approximately 2000 non-native control agents have been made against arthropod pests in 196 countries or islands with remarkably few environmental problems. Biological control is a key component of a 'systems approach' to integrated pest management, to counteract insecticide-resistant pests, withdrawal of chemicals and minimize the usage of pesticides. Current studies indicate that genetically modified insect-resistant Bt crops may have no adverse effects on the activity or function of predators or parasitoids used in biological control. The introduction of rational approaches for the environmental risk assessment of non-native control agents is an essential step in the wider application of biological control, but future success is strongly dependent on a greater level of investment in research and development by governments and related organizations that are committed to a reduced reliance on chemical control.