Causality of mesothelioma: SV40 question.
The fragility of the evidence for SV40 association with human cancer is seen in studies of NHL. A publication in 1999 stated that SV40 is rarely present in NHL. In 2002, two laboratories reported SV40 sequences in 42% to 43% of cases of NHL . One of these laboratories also detected SV40 sequences in small proportions of pediatric tumors (e.g., Wilm's tumor, hepatoblastoma, rhabdomyosarcoma, medulloblastoma, osteosarcoma, and retinoblastoma) and adult carcinomas (e.g., lung, colon, breast, and prostate) These positive results were not confirmed in subsequent studies published in 2003. Capello et al and Mackenzie et al failed to detect SV40 sequences in NHL tissues. Sanjose et al examined sera from patients with NHL and from controls for antibodies reactive to SV40 VLPs, and they detected no significant differences between the two groups. The association of SV40 with NHL is in doubt. An etiologic link between a virus and a cancer becomes plausible when evidence from different lines of enquiry (e.g., epidemiology, pathogenesis, and molecular mechanisms) is mutually reinforcing and together provides a coherent picture that can connect the biology the virus to the characteristics of the disease. The associations of human papillomaviruses with cervical cancer and hepatitis B and C viruses with hepatocellular carcinoma are examples in which the etiologic link is clear. With SV40 and mesothelioma, the data on viral sequences in tumors is inconsistent and disputed, and serologic evidence does not support any association. The epidemiologic data do not show that documented exposures tt SV40 increase the risk of mesothelioma. It seems improbable that a single virus (which cannot be conclusively demonstrated to be present in the community) contributes to the development of such a wide variety of tumors, spanning all age groups and histologic types. The weaknesses in the evidence linking SV40 with mesothelioma are summarized in Box 11 It seems unlikely that infection with SV40 contributes to the development of human mesothelioma or any other human cancer.


The membrane-bound cell division protein DivIB is localized to the division site in Bacillus subtilis.
The cell division gene divIB of Bacillus subtilis is essential for the normal rate of growth and division. The gene product, DivIB, is a membrane-bound protein in which the bulk of the protein (at the C-terminal end) is on the exterior surface of the cell membrane. DivIB is involved in the early stages of septum formation, but its exact role in cell division is unknown. To gain more information about the mode of action of DivIB in septum formation, we determined the location of DivIB within the cell membrane using immunofluorescence. This immunolocalization approach established that DivIB becomes localized to the division site before visible septation and remains localized to this site throughout the division process. Various DivIB immunostaining patterns were observed in immunofluorescence experiments and, together with cell length and nucleoid distance measurements, have allowed us to propose two models to describe DivIB localization during the cell cycle.


[Use of molecular-biological methods for identifying brucella in a comparative analysis of strains, isolated from sick dogs].
Ten strains isolated from sick dogs in 1998 in St. Petersburg were studied by traditional and molecular biological methods of Brucella identification. PCR study confirmed that the isolated cultures were Brucellae, and comparative study of the traditional phenotypical characteristics and protein and antigenic composition allowed referring all the isolated strains to B. canis. Traditional identification showed similarity of 7 strains with the reference B. canis strain RM6/66, and 3 strains were similar to B. canis Mex 51 strain. These results confirmed the division of B. canis into two biovars. Polyacrylamide gel electrophoresis with sodium dodecyl sulfate demonstrated the identity of protein profiles of 10 strains isolated from dogs to the reference B. canis RM6/66 strain. Immunoblotting analysis with S- and R-specific rabbit antisera also demonstrated the identity of antigens binding IgG antibodies in the strains isolated from dogs to the reference B. canis RM6/66 strain.


Effects of thermal stress on human performance.
Experimental evidence indicates that even relatively mild thermal stress may affect human performance. Tasks requiring manual dexterity and muscular strength are clearly impaired by cold exposure, while decrements in vigilance performance and endurance are well documented effects of heat stress. The considerable variation in results regarding the effects of thermal stress may, to some extent, be attributable to complex interactions between exposure conditions, task characteristics, and individual factors. In the present paper the relevance of some of the earlier research work on heat and cold stress is evaluated in the light of the practical and theoretical implications of more recent findings. Current work regarding the nature and extent of the effects of thermal stress on more complex performance is discussed. Attention is also focused on the significance of individual skill and training experience for performance under unfavorable conditions.


Regulation of methanol dehydrogenase synthesis in Paracoccus denitrificans.
The region downstream from the methanol dehydrogenase (MDH) structural gene has been cloned and sequenced. MDH promoter activity have been studied by using a broad-host-range promoter probe vector.


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Corynebacterium group D2 pyelonephritis.
We believe this represents the first known case of Group D2 pyelonephritis in a seventy-two-year-old white man. Previous to this report D2 organisms have been associated only with alkaline-encrusted cystitis and struvite stones in urology.


Changes in the properties of calcium-carbon inclusions during leaf development and their possible relationship with leaf functional maturation in three inclusion-bearing species.
In many plant species, carbon-calcium inclusion (calcium oxalate crystals or cystoliths containing calcium carbonate) formation is a fundamental part of their physiology even necessary for normal growth and development. Despite the long-standing studies on carbon-calcium inclusions, the alterations in their properties during leaf development and their possible association with the maturation of the photosynthetic machinery have not been previously examined. In order to acquire more insights into this subject, we examined three of the most common species bearing abundant inclusions of different types, i.e., Amaranthus hybridus, Vitis vinifera, and Parietaria judaica. Results of our study showed that, irrespective of species and type of inclusion, similar patterns in the alterations of their properties are observed during leaf maturation, except for some differences in cell differentiation and distribution between raphides and druses in Vitis vinifera. As expected, inclusion formation has taken place at very early developmental stages and maximum density was observed in very young leaves. Inclusion properties are changing in a coordinated way with leaf area and these modifications are compatible with the concept that each idioblast or lithocyst "services" a finite number/area of adjacent cells. This tight coordination is also evident at the whole leaf level. Moreover, we observed an association of the properties of carbon-calcium inclusions and gas exchange, suggesting a possible implication of these structures in photosynthesis.


A new strategy for large-scale preparation of high-titer recombinant adeno-associated virus vectors by using packaging cell lines and sulfonated cellulose column chromatography.
The extensive testing of adeno-associated virus (AAV) as a vector for human gene therapy has been hampered by low efficiency of the current packaging system, which is based on transient transfection with plasmid DNAs and infection with adenovirus in permissive cells. In an effort to resolve this problem, HeLa cell-based packaging cell lines were established. These packaging cells carry multiple copies of the AAV genome lacking the inverted terminal repeat (ITR) sequences. The AAV genes were silent in these cells but inducibly expressed by adenovirus infection. When the AAV vector plasmid containing the neoR gene flanked by the ITRs was also integrated into these cells, efficient production of the recombinant AAV particles occurred after adenovirus infection. AAV vector particles in cell lysates could be concentrated by sulfonated cellulose column chromatography. Using the packaging cells and the column chromatography technique, it is possible to prepare AAV vectors with the titer of higher than 10(8) cfu/ml or 5 x 10(10) particles/ml. This new strategy should be useful for testing AAV vectors in vivo.


A profile of the inactive nurse. One solution to the shortage?
The reasons why short-staffed hospitals should consider attracting inactive nurses back to practice are clear: they are an excellent potential source of staff, and there are nearly 400,000 of them. How to attract them back, however, is not as evident. A North Carolina health-education agency surveyed approximately 430 inactive nurses, asking them what made them leave and what would bring them back.


Mercury in wild mushrooms and underlying soil substrate from Koszalin, North-central Poland.
Concentrations of total mercury were determined by cold-vapour atomic absorption spectroscopy (CV-AAS) in 221 caps and 221 stalks of 15 species of wild growing higher fungi/mushrooms and 221 samples of corresponding soil substrate collected in 1997-98 in Manowo County, near the city of Koszalin in North-central Poland. Mean mercury concentrations in caps and stalks of the mushroom species examined and soils varied between 30+/-31 and 920+/-280, 17+/-11 and 560+/-220, and 10+/-9 and 170+/-110 ng/g dry matter, respectively. Cap to stalk mercury concentration quotients were from 1.0+/-0.4 in poison pax (Paxillus involutus) to 2.8+/-0.7 in slippery jack (Suillus luteus). Brown cort (Cortinarius malicorius), fly agaric (Amanita muscaria), orange-brown ringless amanita (A. fulva), red-aspen bolete (Leccinum rufum) and mutagen milk cap (Lactarius necator) contained the highest concentrations of mercury both in caps and stalks, and mean concentrations varied between 600+/-750 and 920+/-280 and 370+/-470 and 560+/-220 ng/g dry matter, respectively. An estimate of daily intake of mercury from mushroom consumption indicated that the flesh of edible species of mushrooms may not pose hazards to human health even at a maximum consumption rate of 28 g/day. However, it should be noted that mercury intake from other foods will augment the daily intake rates. Species such as the sickener (Russula emetica), Geranium-scented russula (R. fellea) and poison pax (P. involutus) did not concentrate mercury as evidenced from the bioconcentration factors (BCFs: concentrations in mushroom/concentration in soil substrate), which were less than 1. Similarly, red-hot milk cap (L. rufus), rickstone funnel cap (Clitocybe geotropa) and European cow bolete (S. bovinus) were observed to be weak accumulators of mercury. Fly agaric (A. muscaria) accumulated great concentrations of mercury with BCFs reaching 73+/-42 and 38+/-22 in caps and stalks, respectively. Mercury BCFs of between 4.0+/-2.3 and 23+/-25 (caps) and 2.6+/-1.9 and 14+/-12 (stalks) were noted for the other mushroom species. Relatively great concentrations of mercury in fly agaric (A. muscaria) were due to preferential uptake of this element by this species.


CB1 and CB2 cannabinoid receptors differentially regulate the production of reactive oxygen species by macrophages.
We investigated the mechanism by which cannabinoid receptors-1 (CB1) and -2 (CB2) modulate inflammatory activities of macrophages. Real-time polymerase chain reaction showed the predominant CB2 expression in freshly isolated human monocytes. PMA, a potent inducer of differentiation, upregulated CB1 and increased CB1:CB2 transcript ratio from 1:17.5 to 1:3 in 5 days of culture. Immunohistochemistry showed that CB1 protein was colocalized in CD68- and CD36-positive macrophages in human atheroma. Through selective expression of CB1 or CB2 to thioglycollate-elicited peritoneal macrophages, we proved that CB1 and CB2 mediate opposing influences on the production of reactive oxygen species (ROS). Flow cytometry showed that cannabinoid-induced ROS production by macrophages was CB1-dependent. Immunoblotting assays confirmed that macrophage CB1, not CB2, induced phosphorylation of p38-mitogen-activated protein kinase, which modulated ROS production and the subsequent synthesis of tumour necrosis factor-alpha and monocyte chemoattractant protein-1. Pull-down assays showed that the Ras family small G protein, Rap1 was activated by CB2. Dominant-negative Rap1 profoundly enhanced CB1-dependent ROS production by macrophages, suggesting CB2 Rap1-dependently inhibits CB1-stimulated ROS production. CB1 promotes pro-inflammatory responses of macrophages through ROS production, which is negatively regulated by CB2 through Rap1 activation. Blocking CB1 together with selective activation of CB2 may suppress pro-inflammatory responses of macrophages.


Local burn versus local cold induced acute effects on in vivo microcirculation and histomorphology of the human skin.
The impact of burns and colds on human skin microcirculation and histomorphology has not been compared as yet. Reflectance confocal microscopy (RCM) enables in vivo insight in human skin on cellular and subcellular levels. We evaluated analogies and differences of thermal injuries on microcirculation and histomorphology in vivo using RCM. Local superficial burn (6 female, 4 male; aged 28.4 ± 2.9 years, burn group) versus superficial cold (4 female, 6 male; aged 30.4 ± 5.2 years, cold group) was induced on the dorsum of the hand in an experimental immersion hand model. In vivo RCM was performed prior (control), immediately (t1) and 15 minutes (t2) following thermal injury to evaluate: Individual blood cell flow (IBCF), functional capillary density (FCD), epidermal thickness (ET), and granular cell size (GCS). In the burn group, IBCF was increased at t1 (78.02 ± 2.60/min) and remained elevated at t2 (84.16 ± 3.04/min). In the cold group, IBCF decreased at t1 (12.62 ± 2.12 min) and increased at t2 (74.24 ± 3.14/min, P < 0.05) compared to the controls (58.23 ± 3.21/min). FCD was 6.74 ± 0.52/mm(2) in controls and increased at both t1 (7.82 ± 0.72/mm(2)) and t2 (8.02 ± 0.81/mm(2)) in the burn group. In the cold group, FCD decreased at t1 (2.60 ± 0.42/mm(2)) and increased at t2 (7.92 ± 0.44/mm(2), P < 0.05). ET increased at both t1 (43.12 ± 4.08 μm, P > 0.05) and t2 (47.26 ± 4.72 μm, P < 0.05) in the burn group. In the cold group, ET decreased at t1 (39.92 ± 3.14 μm, P > 0.05) and increased at t2 (44.72 ± 4.06 μm, P < 0.05) compared to the controls (41.26 ± 3.82 μm). Control GCS was 726.9 ± 59.4 μm(2) and increased at both t1 (739.8 ± 69.8 μm(2), P > 0.05) and t2 (762.6 ± 71.4 μm(2), P < 0.05) in the burn group. In the cold group, GCS decreased at t1 (712.4 ± 53.8 μm(2), P > 0.05) and increased at t2 (742.6 ± 64.8 μm(2), P < 0.05). Superficial burn induces more cellular destruction and cold leads to huge fluctuation in tissue perfusion, however, with moderate impact on histomorphology. The effect on dermal capillaries suggests a selective neural control and cold injuries might down-regulate this system, much more than burns can activate it.


Successful shade matching--what does it take?
Dental practitioners can avoid expensive and embarrassing shade matching errors by learning about the structure and function of the eye and how to control the external and internal factors that affect human vision. The color of objects is identified by the visible light that they reflect; objects can reflect only the light that shines upon them. Eyes sense the hue, intensity, and saturation of light, so lighting of proper intensity and color balance is necessary for consistent, correct shade-matching. Although sunlight is the most common standard for good lighting, in everyday practice, it cannot be used for shade matching because of its extreme variability with weather conditions, time of day, and season of the year. In addition to external factors such as lighting, conditions internal to the observer also play a role in accurate color perception. Color vision confusion (CVC), often called "color blindness," is a serious handicap for dental professionals. The popular assumption is that very few people are afflicted; in fact, all people suffer from some level of color vision confusion. This distortion of visual information can be permanent or transitory. Therefore, controlling these variables can lead to color matching success.


A decomposition model to track gene expression signatures: preview on observer-independent classification of ovarian cancer.
A number of algorithms and analytical models have been employed to reduce the multidimensional complexity of DNA array data and attempt to extract some meaningful interpretation of the results. These include clustering, principal components analysis, self-organizing maps, and support vector machine analysis. Each method assumes an implicit model for the data, many of which separate genes into distinct clusters defined by similar expression profiles in the samples tested. A point of concern is that many genes may be involved in a number of distinct behaviours, and should therefore be modelled to fit into as many separate clusters as detected in the multidimensional gene expression space. The analysis of gene expression data using a decomposition model that is independent of the observer involved would be highly beneficial to improve standard and reproducible classification of clinical and research samples. We present a variational independent component analysis (ICA) method for reducing high dimensional DNA array data to a smaller set of latent variables, each associated with a gene signature. We present the results of applying the method to data from an ovarian cancer study, revealing a number of tissue type-specific and tissue type-independent gene signatures present in varying amounts among the samples surveyed. The observer independent results of such molecular analysis of biological samples could help identify patients who would benefit from different treatment strategies. We further explore the application of the model to similar high-throughput studies.


Seasonal variation in immune measurements and MRI markers of disease activity in MS.
The exact mechanisms by which T cells contribute to MS progression are not known. Recently, the results of cross-sectional studies suggested seasonal variation of both interferon (IFN)-gamma production and the number of active MRI lesions in MS. To investigate whether seasonal fluctuations of IFN-gamma and active MRI lesions could be confirmed and whether any correlations could be detected. Data were analyzed from a group of 28 MS patients in whom detailed longitudinal monitoring of both immune function and MRI measurements had taken place. Significant seasonal variation was observed in T-cell activation as measured by the ability of T cells to secrete the pro-inflammatory cytokines tumor necrosis factor-alpha and IFN-gamma. Maximum values were found in samples obtained during autumn. Even though clear fluctuations were observed, no significant seasonal variation could be detected in the number of active MRI lesions. Fluctuations of in vitro IFN-gamma secretion correlated weakly with changes in active MRI lesions. The finding of seasonal variation of immune function in serially MRI-monitored MS patients suggests an environmental role in T-cell activation.


Production and Assay of Recombinant Multisubunit Chromatin Remodeling Complexes.
We have developed a novel system to facilitate the rapid and easy cloning of multiple genes (>10) in under a week. Using this system we have been able to successfully clone, overexpress, and purify a number of large multimeric proteins from insect cells, including the chromatin remodeling complexes SWR1 and INO80. Using Förster resonance energy transfer (FRET)-based assays we have demonstrated that our overexpressed enzymes have activities comparable to those purified from sources where the proteins are expressed under their endogenous promoters.


Ineffectiveness of intravenous ascorbic acid as an acidifying agent in man.
The acidifying effect of intravenous (IV) ascorbic acid was studied in seven healthy adult volunteers. After obtaining baseline urine and blood samples, 2-g IV doses of ascorbic acid were administered to each subject during a 20-minute period. Venous blood samples were obtained at times 0.5, 1, and 2 hours, and urine was collected at times 0.5, 1, 2, and 3 hours. Our results show that venous blood pH, plasma bicarbonate concentration, urine PCO2, and urine bicarbonate excretion did not change significantly during the study period. Urinary titratable acidity, ammonium excretion, and net hydrogen ion excretion decreased, and urinary pH actually showed a significant rise at two hours. We therefore conclude that IV ascorbic acid administered in recommended doses does not effectively acidify urine.


Furosemide: progress in understanding its diuretic, anti-inflammatory, and bronchodilating mechanism of action, and use in the treatment of respiratory tract diseases.
Accumulated experimental and clinical data suggest that adrenocorticosteroids and/or endogenous ouabain-like substances may play an important role in the mechanism of furosemide diuretic action. It was reported that the drug is highly bound in the adrenals, lungs, kidney, spleen, and liver. In patients with liver cirrhosis, furosemide exerted a markedly decreased natriuretic effect compared with normal subjects, and the plasma levels of circulating endothelin and atrial natriuretic factor (ANF) were significantly elevated. In neonates, after administration of furosemide, the urinary excretion of endothelin-1 and aldosterone increased markedly, and it is known that endothelin may release ANF and aldosterone in a dose-dependent manner. Furosemide was used to stimulate zona glomerulosa, whereas ANF decreased the production of steroids in zona glomerulosa and fasciculata cell culture owing to stimulation by various factors. Because the concomitant use of ANF and furosemide appeared to be diuretically effective in newborns after cardiac surgery, one may suggest that furosemide competes with ANF for its effects on the adrenals. Furosemide administered by inhalation exerted a protective effect on allergic and perennial nonallergic rhinitis and was effective in preventing the postsurgical recurrence of nasal polyposis. The drug can also be used as an antiasthmatic agent. In preterm ventilator-dependent infants with chronic lung disease, aerosolized furosemide improved pulmonary function with no marked effect on diuresis. In adults and children with asthma, furosemide exerted a protective effect against bronchoconstriction induced by several indirect stimuli similar to that of disodium cromoglycate or nedocromil. Aerosolized furosemide had a preventive effect also on bronchoconstriction induced by inhaled lysine acetylsalicylate in patients with aspirin-sensitive asthma. In high-dose beclomethasone-dependent asthma, inhaled lysine acetylsalicylate and furosemide exerted a mutually potentiating antiasthmatic activity, allowing considerable sparing of the inhaled steroid. It is proposed that this effect may be explained by the corticosteroid-sparing action of lysine released from the lysine acetylsalicylate molecule because similar beneficial effects were also obtained after the concomitant use of epsilon-aminocaproic acid (whose chemical structure is almost the same as that of lysine) and prednisone. Furosemide exhibited an anti-inflammatory effect through inhibition of production and release of cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor-alpha from peripheral mononuclear cells, which may have a beneficial effect on local inflamed tissue imbalance in the ratio of different cytokines, thus improving the sensitivity of target cells to endogenous glucocorticosteroids.


[Experiences with the aerial distribution of baits for the oral immunization of foxes against rabies in eastern Germany].
Since the first test in spring 1990, aerial distribution of baits has been developed as the main bait distribution variant in the new federal states of Germany. Experiences are given in respect of organisation, management and enforcement of oral immunization campaigns and cooperation between veterinary agencies and flight enterprises. Practical aspects are described to ensure bait density, preparation of maps, creation of technical preconditions and realisation of dropping rhythm. Aerial dropping in comparison with hand distribution of baits was shown to be equivalent at least in respect of bait uptake and seroconversion rate.


Towards a harmonized method for the global reconnaissance of multi-class antimicrobials and other pharmaceuticals in wastewater and receiving surface waters.
Antimicrobial resistance is a worldwide problem that is both pressing and challenging due to the rate at which it is spreading, and the lack of understanding of the mechanisms that link human, animal and environmental sources contributing to its proliferation. One knowledge gap that requires immediate attention is the significance of antimicrobial residues and other pharmaceuticals that are being discharged from wastewater treatment plants (WWTPs) on the dissemination of antimicrobial resistance in the environment. In this work we provide an approach to develop a harmonized analytical method for 8 classes of antimicrobials and other pharmaceuticals that can be used for global monitoring in wastewater and receiving waters. Analysis of these trace organic chemicals in the influent and effluent wastewater, and in the respective upstream and downstream receiving waters from different countries across the globe is not trivial. Here, we demonstrated that sample preparation using solid-phase extraction (SPE) not only provides a convenient and cost-effective shipping of samples, but also adds stability to the analytes during international shipping. It is important that SPE cartridges are maintained at cold temperature during shipment if the duration is longer than 7 days because a significant decrease in recoveries were observed after 7 days in the cartridges stored at room temperature, especially for sulfonamides and tetracyclines. To compensate for sample degradation during shipment, and matrix effects in liquid chromatography/mass spectrometry, the use of stable isotope labeled compounds should be employed when available and affordable. The importance of applying a defined tolerance for the ion ratios (Q/q) that have been optimized for wastewater and surface water is discussed. The tolerance range was set to be the mean Q/q of the analyte standard at various concentrations ±40% for the influent, and ±30% for the effluent, upstream, and downstream samples; for tetracyclines and quinolones, however, the tolerance range was ±80% in order to minimize false negative and false positive detection. The optimized procedures were employed to reveal differences in antimicrobial and pharmaceutical concentrations in influent, effluent, and surface water samples from Hong Kong, India, Philippines, Sweden, Switzerland, and United States. The antimicrobials with the highest concentrations in influent and effluent samples were ciprofloxacin (48,103 ng/L, Hong Kong WWTP 1) and clarithromycin (5178 ng/L, India WWTP 2), respectively. On the other hand, diclofenac (108,000 ng/L, Sweden WWTP 2), caffeine (67,000 ng/L, India WWTP 1), and acetaminophen (28,000 ng/L, India WWTP 1) were the highest detected pharmaceuticals in the receiving surface water samples. Hong Kong showed the highest total antimicrobial concentrations that included macrolides, quinolones, and sulfonamides with concentrations reaching 60,000 ng/L levels in the influent. Antidepressants were predominant in Sweden, Switzerland, and the United States.


Antiproliferative effect of isolated isoquinoline alkaloid from Mucuna pruriens seeds in hepatic carcinoma cells.
The present study was undertaken to investigate the antiproliferative action of isolated M1 (6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid) from Mucuna pruriens seeds using human hepatic carcinoma cell line (Huh-7 cells). Initially, docking studies was performed to find out the binding affinities of M1 to caspase-3 and 8 enzymes. Later, cytotoxic action of M1 was measured by cell growth inhibition (MTT), followed by caspase-3 and 8 enzymes assay colorimetrically. Our results collectively suggested that M1 had strong binding affinity to caspase-8 in molecular modelling. M1 possessed antiproliferative activity on Huh-7 cells (EC50 = 13.97 μM) and also inhibited the action of caspase-8 enzyme, signified process of apoptosis. M1 was active against Huh-7 cells that may be useful for future hepatic cancer treatment.


Histone deacetylases: target enzymes for cancer therapy.
Epigenic regulation of gene transcription has recently been the subject of a fast growing interest particularly in the field of cancer. Enzymatic acetylation and deacetylation of the epsilon-amino groups of lysine residues from nucleosomal histones, represents major molecular epigenic mechanisms controlling gene expression. Histone deacetylases (HDACs) and histone acetyl transferases (HAT) represent the two families of enzymes in charge of the control of the level of acetylation of the histone tails. By removing the acetyl groups that abrogate the positive charge of the lysine residues that maintain the histone tails attached to DNA, HDACs repress transcription. In mammals, these latter enzymes form three groups of related enzymes based on their sequence homology and are classified as HDACs I, II and III. Global inhibition of the HDACs I and II groups results in cell growth arrest and apoptosis of cancer cells and alters tumor growth in in vivo experimental models. Their surprisingly low general toxicity and their impressive efficiency in preclinical cancer models has led to consider HDAC inhibitors as very promising new anticancer pharmacological agents. In this review, we attempt to give a comprehensive overview of the role and the involvement of HDAC in carcinogenesis as well as the current progress on the development of HDAC general and specific inhibitors as new cancer therapies.


Impacts of human activities on nutrient transport in the Yellow River: The role of the Water-Sediment Regulation Scheme.
Anthropogenic activities alter the natural states of large rivers and their surrounding environment. The Yellow River is a well-studied case of a large river with heavy human control. An artificial managed water and sediment release system, known as the Water-Sediment Regulation Scheme (WSRS), has been carried out annually in the Yellow River since 2002. Nutrient concentrations and composition display significant time and space variations during the WSRS period. To figure out the anthropogenic impact of nutrient changes and transport in the Yellow River, biogeochemical observations were carried out in both middle reaches and lower reaches of the Yellow River during 2014 WSRS period. WSRS has a direct impact on water oxidation-reduction environment in the middle reaches; concentrations of nitrite (NO2-) and ammonium (NH4+) increased, while nitrate (NO3-) concentration decreased by enhanced denitrification. WSRS changed transport of water and sediment; dissolved silicate (DSi) in the middle reaches was directly controlled by sediments release during the WSRS while in the lower reaches, DSi changed with both sediments and water released from middle reaches. During the WSRS, the differences of nutrient fluxes and concentrations between lower reaches and middle reaches were significant; dissolved inorganic phosphorous (DIP) and dissolved inorganic nitrogen (DIN) were higher in low reaches because of anthropogenic inputs. Human intervention, especially WSRS, can apparently change the natural states of both the mainstream and estuarine environments of the Yellow River within a short time.


Nitric oxide inhibition of platelet deposition on biomaterials.
Platelet adhesion and aggregation restrict the clinical applicability of blood-contacting biomaterials. Nitric oxide (NO) is a simple biological molecule that may be incorporated into biomaterials to inhibit platelet deposition. The toxicity of NO at superphysiological levels necessitates the determination of aqueous NO concentrations and fluxes that effectively inhibit platelet deposition. In this study, a novel NO delivery device has been developed to study NO inhibition of platelet deposition in a dynamic in vitro environment. Gaseous NO was delivered via a semipermeable membrane to a radiolabeled platelet suspension perfusing a thin flow slit. The membrane was coated with a platelet-agonistic protein. Spatial NO flux and concentration profiles in the flow slit are predictable using a mathematical model. Platelet inhibition was essentially complete at 0.1 ppm gaseous NO exposure, corresponding to a surface concentration of 0.09 nM and surface fluxes between 0.3 and 0.6 femtomoles cm-2s-1. These threshold values of NO exposure for significant platelet inhibition were unchanged irrespective of the platelet agonist, perfusion times, or shear rates. At lower NO exposures (0.02 ppm), platelet inhibition was only partial with the degree of inhibition dependent on the nature of the agonistic protein. This study yields information useful towards the design and development of biomaterials incorporating NO for the reduction of platelet-biomaterial interactions.


Teaching instrument: a laparoscopic training model.
A laparoscopic training model with female surface anatomy has been developed. This training model is made of a plastic boutique-showing model that is equivalent to normal-size female anatomy from the neck to the upper thigh. Four holes were made on the model's abdominal wall as puncture-sites to enter the cavity, the first is 80-mm diameter at the umbilical area, and the other three 38-mm diameter holes are located on both sides of the lower abdomen and suprapubic area. The umbilical hole can be covered with a simulated abdominal wall made from 6.5-mm insulation sheet, fixed to the model using a rubber band. The other three puncture-sites were plugged with a flexible rubber diaphragm as working ports. When used as video-laparoscopy, the auto-focus camcorder is used as a telescope and is connected to a regular television set as a monitor. This model can be used for training of abdominal entry by Veress needle or trocar, laparoscopic tubal ligation (LTL), and video-eye-hand co-ordination. This model has been a training medium in our Department for 1 year and was included in the OSCE for the Board Examination of the Royal Thai College of Obstetricians and Gynecologists in the year 2000 to assess the process of Veress needle insertion.


[The evolution of ocular refraction after laser excimer surgery--PRK and LASEK techniques].
The purpose of the refractive surgery is to diminish the refractive errors and to obtain a better visual acuity without correction. The study presents the evolution of the refraction after two methods of excimer laser refractive surgery (PRK and LASEK). The results show that these surgical methods are safe and predictable when there are correct indications and well performed interventions.


[Efficacy of prevention of periodontal diseases].
In the present study the results of individual programmes of dental care conducted in a selected population of young adults were shown. The periodontal status was recorded using GPM/T, CPITN, QHI index criteria. In the period of 18 months there was no progression of gingivitis. The severity of gingivitis was clearly reduced, but the percentage distribution was not influenced. The results of the study indicate directions for prevention, diagnosis and therapy. In order to reduce the incidence of the diseases dental care should be concentrated on the motivation and instruction of oral hygiene, plaque control and removal of dental calculus. The findings of the study confirm that a prophylactic programme can prevent more severe gingivitis and marginal periodontitis.


Absence of gender effects on attention deficit hyperactivity disorder: findings in nonreferred subjects.
In a previous study, the authors found that, compared with referred boys with attention deficit hyperactivity disorder (ADHD), girls are less likely to manifest comorbid disruptive behavior disorders and learning disabilities--characteristics that could adversely affect identification of ADHD in girls. However, because referral bias can affect outcome, these findings require replication in nonreferred groups of ADHD subjects. The authors evaluated gender effects in a large group of nonreferred siblings (N=577) of probands with ADHD and non-ADHD comparison subjects. Ninety-eight of the nonreferred siblings (N=73 males, N=25 females) met the criteria for diagnosis of ADHD, and 479 (N=244 males, N=235 females) did not meet those criteria. All siblings were systematically and comprehensively assessed with measures of emotional, school, intellectual, interpersonal, and family functioning. The assessment battery used for the siblings was the same as that used for the probands. The nonreferred males and females with ADHD did not differ in DSM-IV subtypes of ADHD, psychiatric comorbidity, or treatment history. They also showed similar levels of cognitive, psychosocial, school, and family functioning. These findings suggest that the clinical correlates of ADHD are not influenced by gender and that gender differences reported in groups of subjects seen in clinical settings may be caused by referral biases.


Nuclear accumulation of full-length and truncated adenomatous polyposis coli protein in tumor cells depends on proliferation.
The adenomatous polyposis coli (APC) tumor suppressor is a nucleocytoplasmic protein. The nuclear accumulation of APC was recently found to vary depending on cell density, suggesting that putative APC function(s) in the nucleus is controlled by the establishment of cell contacts. We report here that the density-dependent redistribution of APC between nucleus and cytoplasm prevails in 6/6 thyroid and colorectal carcinoma cell lines. Moreover, mutated APC lacking known nuclear localization sequences had the similar distribution pattern as the full-length protein. APC invariably accumulated in the nuclei of Ki-67 expressing cells, but was largely cytoplasmic when cell cycle exit was induced by serum starvation or at high cell density. APC colocalized with beta-catenin in the nucleus only in one cell line (SW480). Also, APC maintained a predominantly nuclear position in early confluent states when cytoplasmic beta-catenin was recruited to newly formed adherens-like junctions. The results indicate that nuclear targeting of APC is driven by cell cycle entry rather than altered cell-cell contact. The ability of C-terminally truncated APC to accumulate in the nucleus suggests that nuclear import signals other than NLS1(APC) and NLS2(APC) are functionally important. Residual function(s) of N-terminal APC fragments in tumor cells carrying APC mutations might be beneficial to tumor growth and survival.


Beta Human Papillomavirus and Associated Diseases.
The cutaneous human papillomavirus (HPV), mostly from β- and γ-HPV genus, is ubiquitously distributed throughout the human body and may be part of the commensal flora. The association of β-HPVs and cutaneous squamous cell carcinoma (cSCC) development was initially reported in patients with the rare genetic disorder Epidermodysplasia verruciformis. Likewise, immunosuppressed organ transplant recipients have an increased susceptibility to β-HPV infections in the skin as well as to cSCC development. Although ultraviolet radiation (UVR) is the main risk factor of cSCC, experimental data points toward β-HPVs as co-carcinogens, which appear to be required solely at early stages of skin carcinogenesis by facilitating the accumulation of UVR-induced DNA mutations. Several epidemiological studies relying on different biomarkers of β-HPV infections have also been conducted in immunocompetent individuals to access their association with cSCC development. Additionally, in vivo and in vitro studies are presenting cumulative evidence that E6 and E7 proteins from specific β-HPVs exhibit transforming activities and may collaborate with different environmental factors in promoting carcinogenesis. Nevertheless, further research is crucial to better understand the pathological implications of the broad distribution of these HPVs.


Engineered UV-A light-responsive gene expression system for measuring sun cream efficacy in mammalian cell culture.
Light-dependent gene regulation systems are advantageous as they allow for precise spatio-temporal control of target gene expression. In this paper, we present a novel UV-A and blue-light-inducible gene control system that is based on the light-dependent heterodimerization of the CRY2 and C1BN domains. Upon their interaction, a transcription factor is released from the cell membrane and initiates target gene expression. Capitalizing on that, sun cream UV-A protection properties were measured intracellularly.


Optimizing economic outcomes in antibiotic therapy of patients with acute bacterial exacerbations of chronic bronchitis.
The social, medical and economic effects of acute bacterial exacerbations of chronic bronchitis on individual patients and the resource implications of this disease for the healthcare sector are considerable. Optimizing the selection of patients who should receive antibiotics according to stringent clinical criteria is the first step in promoting good clinical practice and cost-effectiveness. Antibiotic efficacy is then the major driver of cost, especially when it reduces the need for hospitalization. Resistance to first-line antibiotics can be expected to increase the risk of treatment failure. Other drivers of cost include non-compliance, which predisposes to therapeutic failure, and the selection of resistant strains. Treatment regimens of short duration, once-daily dosing and good tolerability are determinants of good compliance and cost savings. The expenses of first-line antibiotics typically account for only a small proportion of the overall costs of healthcare and the cheapest antibiotics are not necessarily the most cost-effective. The clinical success rate of first-line therapy is the primary determinant of the overall expenditure on healthcare because of the high costs associated with treatment failure, especially if it leads to hospitalization. Factors such as poor patient compliance and high antibiotic resistance rates, which undermine the clinical efficacies of first-line therapy, will increase the overall costs of treatment.


Emergence of population growth models: fast migration and slow growth.
We present aggregation and emergence methods in large-scale dynamical systems with different timescales. Aggregation corresponds to the reduction of the dimension of a dynamical system which is replaced by a smaller model for a small number of global variables at a slow timescale. We study the couplings between fast and slow dynamics leading to the emergence of global properties in the aggregated model. First, we study the case of a single population in a patchy environment. Growth rates are assumed to be linear on each patch. Individuals can migrate from one patch to another at a fast timescale. We choose different density dependent migration processes. In each case, we use aggregation methods to obtain the corresponding growth equation for the total density of the population at a slow timescale. We look for particular density dependent migration processes leading to an aggregated logistic-like equation. Second, we study the case of two interacting populations. A particular choice of density dependent migrations leads to an aggregated competition model.


Conserved epitopes in human and mouse tissue-nonspecific alkaline phosphatase. Second report of the ISOBM TD-9 workshop.
A panel of 19 monoclonal antibodies (MAbs) against human tissue-nonspecific (liver/bone/kidney) alkaline phosphatase (TNAP) was obtained through the ISOBM TD-9 workshop. In the present study, the reactivity of these MAbs has been characterized against mouse TNAP. A mouse embryonic stem cell line, frozen sections of long bones, alkaline phosphatase extracted from mouse bone, and serum were used as the source of TNAP for individual assays. Each MAb was tested for immunoreactivity to mouse TNAP by Western blot analysis, immunohistochemistry and enzyme immunoassay. Antibodies 314 and 315 reacted strongly with mouse TNAP in Western blots, while all other antibodies were negative. By immunohistochemistry, antibodies 314, 315 and 333 produced strong positive staining using frozen sections, while antibody 334 was moderately positive. Enzyme immunoassays indicated that MAb 333 was also able to bind to serum TNAP. These antibodies represent very useful reagents to study the pathophysiological expression of TNAP in mouse tissues and in mouse serum.


[Sources of the cerebrospinal afferent innervation of the pig thyroid gland].
Comparing results of extirpation of the thyroid gland with those obtained at cutting tissues without extirpation of the gland in 1.5--2-month-old pigs of both sex, it was possible to reveal that there are numerous sources of afferent innervation of the thyroid gland in these animals. They include the distal ganglion of the vagus nerve and the cerebrospinal ganglia C1-C6. An essential amount of cells participating in the innervation belong to the distal ganglia of the vagus nerve (main sources), other ganglia concentrate but a small amount of similar neurons (additional sources). A considerable part of neurons participating in the gland innervation are cells of B and C types.


Intermittency of visual information and the frequency of rhythmical force production.
The authors examined the influence of intermittent (40-5,000 ms) visual information on the control of rhythmical isometric force output (0.5, 2.0, and 4.0 Hz) in 10 participants. Force variability decreased as a function of less intermittent visual information only in the 0.5- and 2.0-Hz tasks. Vision influenced the frequency structure of force output through 0-12 Hz in the 0.5-Hz task, but in only the 10.0- to 12.0-Hz range in the 2.0-Hz task and not in the 4.0-Hz task. The effective use of intermittent visual information in force output was mediated by task frequency, and that mediation was reflected in the differential emphasis of feedback and feedforward processes over multiple timescales of control.


A multi-disciplinary approach to understanding the impacts of mines on traditional uses of water in Northern Mongolia.
Mongolia is an example of a nation where the rapidity of mining development is outpacing capacity to manage the potential land and water resources impacts. Further, Mongolia has a particular social and economic reliance on traditional uses of land and water, principally livestock herding. While some mining operations are setting high standards in protecting the natural resources surrounding the mine site, others have less incentive and capacity to do so and therefore are having adverse effects on surrounding communities. The paper describes a case study of the Sharyn Gol Soum in northern Mongolia where a range of mining types, from artisanal, small-scale mining to a large coal mine, operate alongside traditional herding lifestyles. A multi-disciplinary approach is taken to observe and attribute causes to the water resources impacts in the area. Surveys of the herding household community, land use mapping, and monitoring the spatial variations in water quality indicate deterioration of water resources. Collectively, the different sources of evidence suggest that the deterioration is mainly due to small-scale gold mining. The evidence included the perception of 78% of the interviewed herders that water quality had changed due to mining; a change in the footprint of small-scale gold mining from 2.8 to 15.2km(2) during the period 1999 to 2015; and pH and sulphate values in 2015 consistently outside the ranges observed at a baseline site in the same region. It is concluded that the lack of baseline data and effective governance mechanisms are fundamental challenges that need to be addressed if Mongolia's transition to a mining economy is to be managed alongside sustainability of herder lifestyles.


Heterozygous SOX9 mutations allowing for residual DNA-binding and transcriptional activation lead to the acampomelic variant of campomelic dysplasia.
Campomelic dysplasia is a malformation syndrome with multiple symptoms including characteristic shortness and bowing of the long bones (campomelia). CD, often lethal due to airway malformations, is caused by heterozygous mutations in SOX9, an SRY-related gene regulating testis and chondrocyte development including expression of many cartilage genes such as type II collagen. Male to female sex reversal occurs in the majority of affected individuals with an XY karyotype. A mild form without campomelia exists, in which sex-reversal may be also absent. We report here two novel SOX9 missense mutations in a male (c.495C>G; p.His165Gln) and a female (c.337A>G; p.Met113Val) within the DNA-binding domain leading to non-lethal acampomelic CD. Functional analyses of mutant proteins demonstrate residual DNA-binding and transactivation of SOX9-regulated genes. Combining our data and reports from the literature we postulate a genotype-phenotype correlation: SOX9 mutations allowing for residual function lead to a mild form of CD in which campomelia and sex reversal may be absent.


The incidence of necrotizing changes in adults with pneumococcal pneumonia.
Necrotizing pneumonia is generally considered a rare complication of pneumococcal infection in adults. We systematically studied the incidence of necrotizing changes in adult patients with pneumococcal pneumonia and examined the severity of infection, role of causative serotypes, and association with bacteremia. We used a database of all pneumococcal infections identified at our medical center between 2000 and 2010. Original readings of chest X-rays (CXR) and computerized tomography (CT) were noted. Images were then independently reevaluated by 2 radiologists. The severity of disease at admission was assessed using SMART-COP and Pneumonia Outcomes Research Team (PORT) scoring systems. In 351 cases of pneumococcal pneumonia, necrosis was reported in no (0%) original CXR readings and in 6 of 136 (4.4%) CTs. With rereading, 8 of 351 (2.3%) CXR and 15 of 136 (11.0%) CT had necrotizing changes. Overall, these changes were identified in 23 of 351 (6.6%) patients. The incidence of bacteremia and the admitting SMART-COP and PORT scores were similar in patients with and without necrosis (P = 1.00, P = .32, and P = .54, respectively). Type 3 pneumococcus was more commonly isolated from patients with necrosis than from patients without necrosis (P = .05), but 10 other serotypes were also implicated in 16 cases for which the organism was available for typing. Necrotizing changes in the lungs were seen in 6.6% of a large series of adults with pneumococcal pneumonia but were often overlooked on initial readings. Patients with necrosis were not more likely to have bacteremia or more severe disease. Type 3 pneumococcus was the most commonly identified serotype.


Natriuretic peptides in the control of lipid metabolism and insulin sensitivity.
Natriuretic peptides have long been known for their cardiovascular function. However, a growing body of evidence emphasizes the role of natriuretic peptides in human substrate and energy metabolism, thereby connecting the heart with several insulin-sensitive organs like adipose tissue, skeletal muscle and liver. Obesity may be associated with an impaired regulation of the natriuretic peptide system, also indicated as a natriuretic handicap. Evidence points towards a contribution of this natriuretic handicap to the development of obesity, type 2 diabetes mellitus and cardiometabolic complications, although the causal relationship is not fully understood. Nevertheless, targeting the natriuretic peptide pathway may improve metabolic health in obesity and type 2 diabetes mellitus. This review will focus on current literature regarding the metabolic roles of natriuretic peptides with emphasis on lipid metabolism and insulin sensitivity. Furthermore, it will be discussed how exercise and lifestyle intervention may modulate the natriuretic peptide-related metabolic effects.


Spiramycin vs. placebo for treatment of acute diarrhea caused by Cryptosporidium.
The efficacy of spiramycin was evaluated in a double blind, placebo-controlled study of 44 immunocompetent infants ages 2 to 13 months who had acute diarrhea caused by Cryptosporidium. Twenty-one patients received spiramycin (100 mg/kg/day) for 10 days and 23 received placebo. On admission the patients in both groups were comparable regarding demographic and clinical characteristics. The infants who were treated with spiramycin had a shorter duration of diarrhea (mean, 5.2 vs. 7.3 days; P = 0.002) and a shorter duration of excretion of oocysts in the stools (7.1 vs. 8.5 days; P = 0.032) compared with those treated with placebo. No clinical or parasitologic relapses were seen in patients of both groups. Mild adverse effects to spiramycin were observed in 2 patients (10%). Spiramycin appeared to hasten clinical recovery and decrease the duration of oocyst excretion in immunocompetent children with diarrheal illness caused by Cryptosporidium.


Polyamines in conventional and organic vegetables exposed to exogenous ethylene.
Relationships between endogenous levels of polyamines by thin layer chromatography (TLC) and gas chromatography (GC), nitrate and response to the application of ethylene were established between organic and conventional vegetables (broccoli, collard greens, carrots and beets), both raw and cooked. Responses to ethylene showed that organic plants were less responsive to the growth regulator. The levels of free polyamines obtained by TLC were higher in organic vegetables. Organic broccoli showed higher levels of putrescine (Put), and cooking resulted in lowering the overall content of these amines. Conventional collard green showed the highest level of putrescine in the leaves compared with organic. Tubers of carrots and beets contain the highest levels of Put. These plants also contain high levels of spermine. GC analysis showed the highest polyamines contents compared with those obtained by TLC. Cooking process decreased putrescine and cadaverine content, both in conventionally and organically grown vegetables. Organic beets contain lower NO3(-) compared with its conventional counterpart.


Interactive visual analysis of drug-target interaction networks using Drug Target Profiler, with applications to precision medicine and drug repurposing.
Knowledge of the full target space of drugs (or drug-like compounds) provides important insights into the potential therapeutic use of the agents to modulate or avoid their various on- and off-targets in drug discovery and precision medicine. However, there is a lack of consolidated databases and associated data exploration tools that allow for systematic profiling of drug target-binding potencies of both approved and investigational agents using a network-centric approach. We recently initiated a community-driven platform, Drug Target Commons (DTC), which is an open-data crowdsourcing platform designed to improve the management, reproducibility and extended use of compound-target bioactivity data for drug discovery and repurposing, as well as target identification applications. In this work, we demonstrate an integrated use of the rich bioactivity data from DTC and related drug databases using Drug Target Profiler (DTP), an open-source software and web tool for interactive exploration of drug-target interaction networks. DTP was designed for network-centric modeling of mode-of-action of multi-targeting anticancer compounds, especially for precision oncology applications. DTP enables users to construct an interaction network based on integrated bioactivity data across selected chemical compounds and their protein targets, further customizable using various visualization and filtering options, as well as cross-links to several drug and protein databases to provide comprehensive information of the network nodes and interactions. We demonstrate here the operation of the DTP tool and its unique features by several use cases related to both drug discovery and drug repurposing applications, using examples of anticancer drugs with shared target profiles. DTP is freely accessible at http://drugtargetprofiler.fimm.fi/.


Isolation, characterization, and in rats plasma pharmacokinetic study of a new triterpenoid saponin from Dianthus superbus.
One new oleanolic acid triterpenoid saponin, 3-O-β-D-glucopyranosyl olean-11, 13(18)-diene-23,28-dioic acid, (hereafter referred to as DS-1) was isolated from the traditional Chinese medicinal plant Dianthus superbus (D. superbus). DS-1 plays an important role in the bioactivity of D. superbus. Thus, a sensitive, reliable and accurate reversed-phased liquid chromatography with tandem mass spectrometry (LC-MS/MS) in negative ion mode was developed and validated for the quantification and pharmacokinetic study of DS-1 in rats plasma. The pharmacokinetic profile showed that DS-1 was rapidly absorbed and eliminated in plasma, indicating that significant accumulation of the compound in biological specimen is unlikely. In addition, poor absorption into systemic circulation was observed after oral administration of DS-1, resulting in low absolute bioavailability (0.92 %).


[Cytokines and severe infections].
In severe infections two factors play a part: the infectious agent and the response of the host. The response of the host involves production of a large number of endogenous mediators including a number of cytokines that are currently the focus of many studies: tumor necrosis factor (TNF alpha), interleukins (IL-1 and IL-6), and interferon gamma (IGN gamma). These cytokines are part of the body's normal defense mechanisms but can have toxic effects when produced in excessive amounts. Although levels of these cytokines are often high in the blood of patients with sepsis, persistence of these elevations is the main indicator of severe infection. Experimentally, injections of TNF alpha and IL-1 reproduce the manifestations of severe sepsis. Mice that are genetically unable to produce TNF alpha are resistant to the injection of endotoxin. Severe sepsis can be prevented by pretreatment of animals with anticytokine agents (polyclonal and monoclonal antibodies and anti-receptor agents ...). Many issues remain unresolved: for instance, the experimental results obtained with an intravenous bolus of endotoxin or bacteria have not been confirmed in some animal models of subacute infection. These models may more closely resemble human infections. The interrelations between these cytokines are extremely complex. Synergistic effects do occur, but the effects of combinations of cytokines can be different from those of each cytokine given alone... It follows that therapeutic use in humans of anti-cytokine molecules is still an approach of uncertain outcome that will perhaps be clarified by ongoing multicenter clinical trials.


Effects of dose and duration of therapy on gentamicin tissue residues in sheep.
The effects of different doses and dosage regimens on gentamicin pharmacokinetics and tissue residues were determined. Five groups of 12 sheep each were given gentamicin IM: group I, 2 mg of gentamicin sulfate/kg once; group II, 6 mg/kg once; group III, 18 mg/kg once; group IV, 6 mg/kg every 24 hours for 3 doses; and group V, 2 mg/kg every 8 hours for 9 doses. Serum concentrations were determined serially until sheep were killed and necropsied. Three sheep from each group were killed at 1, 4, 8, and 12 days after the last dose was administered. Renal cortex, renal medulla, liver, spleen, lung, skeletal muscle, and skeletal muscle at the injection site were assayed for gentamicin. An exponential equation was fitted to the serum concentrations, and various pharmacokinetic variables were determined. Serum clearance tended to increase as the single dose increased (P = 0.0588). Steady-state volume of distribution increased as the single dose was increased (P less than 0.05). Renal cortex contained the highest concentration of gentamicin which decreased in a biexponential manner. Concentrations in all tissues, except the injection site, were dependent upon the amount of the total dose, not the size of the injected dose (P less than 0.05). Concentrations at the injection site were up to 29 micrograms/g of tissue at 1 day after the last dose was given and were dependent upon the amount of total dose from multiple injections, not on the amount of each injected dose (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Optical lectin based biosensor as tool for bacteria identification.
Biosensor techniques are based on biospecific interaction between the biological parts of biosensor with the analyte. In biosensor construction, antibodies are usually used for the detection of analytes such as microorganism, because of very strong and highly specific interaction. The disadvantages of this assay are a long time needed for antibody isolation and purification as well as difficult regeneration of biosensor chip. The use of lectins instead of antibodies could solve these problems because a several hundred lectins are commercially available and their stability in standard buffers is better compared to monoclonal antibodies. While antibody can only be used to detect that antigen it was designed for, lectin as low affinity molecule may bind several different pathogens. Using the discriminative effect of an artificial neural network the application of a lectin array will compensate for the lower specificity. Microbial surfaces bear many of the sugar residues capable of interacting with lectins. The ability of lectins to react with microbial glycoconjugates means that it is possible to employ them as probes and sorbents for whole cells, mutants and numerous cellular constituents and metabolites, and it makes them useful tools for identification or typing of bacteria. Lectins are attractive reagents for the clinical diagnostic laboratory because of their diverse specificity, commercial availability, a wide range of molecular weights, and their stability in standard buffers. The construction of lectin biosensor could be an advantage method for detection of pathogenic bacteria.


Antiparkinsonian actions of blockade of NR2B-containing NMDA receptors in the reserpine-treated rat.
Current symptomatic treatment for Parkinson's disease is based largely on dopamine-replacing agents. The fact that long-term treatment with these drugs is characterized by many side effects has lead to widespread interest in nondopaminergic therapies. To date, however, it has proved difficult to devise a nondopaminergic therapy with significant antiparkinsonian efficacy when administered as monotherapy. Overactivity of the striatolateral pallidal pathway, the "indirect" striatal output pathway, is thought be responsible for the generation of parkinsonian symptoms. Indeed, it has been suggested that selective reduction in the activity of the "indirect" pathway may be achieved by blockade of NR2B-containing NMDA receptors. In the present study, we demonstrate that selective blockade of NR2B-containing NMDA receptors with the polyamine antagonists ifenprodil and eliprodil causes a significant increase in locomotor activity in the reserpine-treated rat model of Parkinson's disease (30 mg/kg ifenprodil, 221.2 +/- 54 mobile counts compared to vehicle, 19.6 +/- 6.87, P < 0.001). Additionally, we show that, subsequent to dopamine depletion, the ability of ifenprodil to bind to the polyamine site and inhibit binding of the NMDA channel blocker [3H] MK-801 is increased fourfold (IC50 3.7 +/- 0.4 microM compared to vehicle, IC50 14.3 +/- 2.34 microM, P < 0.01). We suggest that ifenprodil selectively targets the polyamine site on overactive NR2B-containing NMDA receptors. Thus, we propose that NR2B-selective NMDA receptor antagonists may prove useful in the treatment of Parkinson's disease.


Clinician attendance and delivery practices at hospital-based vaginal deliveries in Western Kenya.
To characterize delivery practices and factors associated with respectful, evidence-based care at a referral hospital in Western Kenya. An exploratory observational study used a standardized birth-observation form to record information on patient characteristics and healthcare practitioner behaviors during uncomplicated vaginal deliveries between June 30, 2014 and July 17, 2014. All deliveries were monitored for whether healthcare staff performed six specific evidence-based practices (three maternal and three neonatal practices). In total, 75 vaginal deliveries were observed. In 48 (64%) deliveries, nursing students were the only practitioners present. The mean number of evidence-based practices performed at each delivery was 3.58. The number of evidence-based practices performed by junior practitioners was higher when a nurse educator was assessing their performance (4.47 vs 3.36, P<0.001). Lower mean respectful-care scores were recorded when delivery teams comprised three or more practitioners (1.38; 95% confidence interval 0.93-1.84 vs 2.74; 95% confidence interval 2.16-3.31, P=0.002). The present study found low rates of evidence-based practice and respectful maternity care; this could serve as a deterrent for women seeking care at the study facility. These findings emphasize the need for a comprehensive approach in increasing the quality of patient care to improve maternal and newborn health outcomes.


Exact alpha-error determination for two-stage sampling strategies to substantiate freedom from disease.
Sampling strategies to substantiate freedom from disease are important when it comes to the trade of animals and animal products. When considering imperfect tests and finite populations, sample size calculation can, however, be a challenging task. The generalized hypergeometric formula developed by Cameron and Baldock (1998a) offers a framework that can elegantly be extended to multi-stage sampling strategies, which are widely used to account for disease clustering at herd-level. The achieved alpha-error of such surveys, however, typically depends on the realization of the sample and can differ from the pre-calculated value. In this paper, we introduce a new formula to evaluate the exact alpha-error induced by a specific sample. We further give a numerically viable approximation formula and analyze its properties using a data example of Brucella melitensis in the Austrian sheep population.


Mertk drives myosin II redistribution during retinal pigment epithelial phagocytosis.
Mertk is a key phagocytic receptor in the immune, male reproductive, and visual systems. In the retinal pigment epithelium, Mertk is required for the daily ingestion of photoreceptor outer segment (OS) tips. Loss of Mertk function causes retinal degeneration in rats, mice, and humans; however, little is known about the mechanism by which Mertk regulates the ingestion phase of retinal pigment epithelial (RPE) phagocytosis. To address this, the authors sought proteins that associated with Mertk during OS phagocytosis. Lysates of RPE-J cells challenged with OS for various times were immunoprecipitated with Mertk antibody. Potential interacting proteins were identified by mass spectrometry and characterized with confocal microscopy, pharmacologic inhibition, and siRNA knockdown coupled with an in vitro phagocytic assay in primary RPE cells. Myh9, the non-muscle myosin II-A heavy chain, was enriched in immunoprecipitates from OS-treated samples. Myosin II-A and II-B isoforms exhibited a striking redistribution in wild-type rat primary RPE cells challenged with OS, moving from the cell periphery to colocalize with ingested OS over time. In contrast, myosin II-A redistribution in response to OS was blunted in primary RPE cells from RCS rats, which lack functional Mertk. Wild-type rat primary RPE cells treated with the myosin II-specific inhibitor blebbistatin or myosin II siRNAs exhibited a significant phagocytic defect. Mertk mobilizes myosin II from the RPE cell periphery to sites of OS engulfment, where myosin II function is essential for the normal phagocytic ingestion of OS.


Hormone dependency in N-nitrosomethylurea-induced rat mammary tumors.
The majority (87%) of N-nitrosomethylurea-induced rat mammary tumors regressed within 1 week after hypophysectomy (hypox). After a hypox-induced tumor regression, ovine PRL (oPRL), and 17 beta-estradiol (E2) were administered separately or in combination in order to define the individual role of these hormones in regulating tumor growth and influencing estrogen (E), progesterone (Pg), and PRL receptor (R) levels. Administration of E2 (2.5 micrograms twice daily) or oPRL (20 IU daily, started 5 days after hypox and continued for 10 days, resulted in stabilization of tumor growth. Simultaneous administration of E2 and oPRL resulted in a synergistic effect and reactivation of tumor growth. ER levels in mammary tumors were significantly lower than those in the control 15 days after hypox (P less than 0.01). Treatment with E2, oPRL, or both simultaneously had no significant effect on ER levels. A significant decline in PgR levels was noted at both 5 and 15 days after hypox. Whereas treatment with oPRL had no significant effect on PgR levels, E2 administration either alone or in combination with oPRL restored PgR levels to control values. PRLR levels were unchanged from control values at 5 days, but significantly declined (P less than 0.005) 15 days after hypox. Treatment with E2, oPRL, or both hormones simultaneously partially maintained PRLR and prevented the decline to the extremely low level noted in the untreated group. We conclude that the growth of nitrosomethylurea-induced rat mammary tumors is dependent on both E2 and PRL. There was a synergistic effect between E2 and PRL on tumor growth but not on ER, PgR, or PRLR. Neither E2 nor PRL had any significant effect on ER after hypox. PgR is under E2 control. Either E2 or PRL or both hormones were able to maintain PRLR in mammary tumors after hypox.


Pre-operative identification of an anomalous left coronary artery from the pulmonary artery in association with tetralogy of Fallot and an aortopulmonary window.
A newborn baby girl was diagnosed with tetralogy of Fallot and an aortopulmonary window. At 3 weeks of age, the coexistent diagnosis of an anomalous left coronary artery from the pulmonary artery was made. Initial surgical intervention included a Takeuchi baffle of the left coronary artery, closure of the aortopulmonary window, and placement of a right Blalock-Taussig shunt. Complete repair was undertaken at 12 months of age. Close pre-operative assessment of the coronary arterial anomaly is crucial in patients with an aortopulmonary window and tetralogy of Fallot.


Facilitating pictorial comprehension with color highlighting.
Pictorials can aid in communicating warning information, but viewers may not always correctly comprehend them. Two experiments focused on whether the use of relevant highlighting could benefit pictorial comprehension. A set of warning-related pictorials were manipulated according to three-color highlighting conditions: highlighting areas more relevant to correct comprehension, highlighting areas less relevant to comprehension, and no highlighting. Participants were asked to describe the purpose and meaning of each pictorial presented to them. The findings from both experiments indicate that comprehension of warning pictorials is higher for the relevant highlighting condition than the other two conditions. The highlighting of less relevant areas reduced comprehension compared to no highlighting. Use of appropriately placed highlighting could benefit the design of a complex symbol by pointing out pertinent areas to aid in determining its intended conceptual meaning.


Serum and urinary markers of exogenous testosterone administration.
In an attempt to find optimal markers of exogenous testosterone (T) administration in male athletes, a number of compounds were measured in 11 healthy men before and after 3, 6 and 9 months of weekly administration of 250 mg i.m. T enanthate and in age-matched untreated controls. The following variables were measured in serum: T, 17 alpha-hydroxyprogesterone (17-OHP), sex hormone-binding globulin (SHBG), estradiol-17 beta, estrone (free + conjugated) and luteinizing hormone (LH). The following variables were measured in urine: T glucuronide (urinary T), epitestosterone glucuronide (urinary epiT), estrone (free + conjugated) and LH. Serum T, serum T/17-OHP ratio, serum T/LH ratio, serum T/SHBG ratio, serum and urinary estrogens, urinary T/creatinine-, T/epiT- and T/LH ratios increased whereas serum 17-OHP, SHBG and LH and urinary epiT/creatinine- and LH/creatinine-ratios decreased significantly during treatment. Levels above the upper reference limit were found in all subjects at 3, 6 and 9 months for serum T/17-OHP and serum and urinary T/LH ratios and at 6 months for the urinary T/epiT ratio. Levels below the lower reference limit were found in all subjects at 3, 6 and 9 months for serum LH and the urinary LH/creatinine ratio, at 3 months for the urinary epiT/creatinine ratio and at 9 months for serum 17-OHP. No other variable showed abnormal values in all subjects at the same occasion. Despite significant changes during treatment, steroid concentrations as such are poor indicators of T doping. Serum and urinary LH levels, T/LH ratios and serum T/17-OHP ratios seem to be the most reliable markers of exogenous T administration in males.


Subzero-temperature stabilization and spectroscopic characterization of homogeneous oxyferrous complexes of the cytochrome P450 BM3 (CYP102) oxygenase domain and holoenzyme.
We describe herein for the first time the formation and spectroscopic characterization of homogeneous oxyferrous complexes of the cytochrome P450 BM3 (CYP102) holoenzyme and heme domain (BMP) at -55 degrees C using a 70/30 (v/v) glycerol/buffer cryosolvent. The choice of buffer is a crucial factor with Tris [tris(hydroxymethyl)aminomethane] buffer being significantly more effective than phosphate. The oxyferrous complexes have been characterized with magnetic circular dichroism spectroscopy and the resulting spectra compared to those of the more well-characterized oxyferrous cytochrome P450-CAM. The formation of a stable substrate-bound oxyferrous CYP BM3 holoenzyme, despite the fact that it has the necessary reducing equivalents for turnover, indicates that electron transfer from the flavin domain to the oxyferrous center is very slow at this temperature. The ability to prepare stable homogeneous oxyferrous derivatives of both BMP and the CYP BM3 holoenzyme will enable these species to be used as starting materials for mechanistic investigation of dioxygen activation.


Acrolein/glutaraldehyde as a fixative for combined light and electron microscopic immunocytochemical detection of pituitary hormones in immersion-fixed tissue.
In an attempt to find a fixative that has a wide range of application in immunocytochemical studies of pituitary tissue, we have investigated the use of acrolein fixation followed by postfixation in osmium. Pituitary glands were excised and immersion-fixed in 2 or 5% acrolein/0.25% glutaraldehyde for 1, 2.5, 5, or 24 hr. Some tissues were subsequently postfixed in 1% OsO4 for 1.5 hr before embedding in Epon-Araldite. Sections were collected for both light and electron microscopic immunocytochemistry, employing several pituitary hormone antisera. Fixation for 2.5 hr with 2% acrolein/0.25% glutaraldehyde followed by osmication resulted in good staining for both light and electron microscopy with all antisera employed. Furthermore, the rapid penetration qualities of this fixative resulted in good ultrastructural preservation throughout the tissue.


Evaluation of the extent and type of bacterial contamination at different stages of processing of cooked ham.
In an attempt to determine the composition and origin of the spoilage flora of refrigerated vacuum-packed cooked ham, the changes in microbial numbers and types were followed along the processing line. Results revealed Lactobacillus sake and Leuconostoc mesenteroides ssp. mesenteroides as the major causative agents of spoilage of sliced ham stored at 4 degrees C and 12 degrees C, due to recontamination in the cutting room. On the contrary, the progressive deterioration of whole ham under the same storage conditions was associated with a non-identifiable group of leuconostoc-like bacteria. Except for lactic acid bacteria, no other organism grew in vacuum packs of either sliced or whole ham. Although atypical leuconostocs could not be detected among isolates recovered from freshly produced whole ham, they appeared to survive cooking and proliferate during storage. Neither these organisms however, nor Lact. sake and Leuc. mesenteroides were important in curing and tumbling as carnobacteria, mainly Carnobacterium divergens, and Brochothrix thermosphacta dominated at this stage. A progressive inversion of the ham microflora from mostly Gram-negative at the beginning of processing to highly Gram-positive prior to cooking was noted. Listeria monocytogenes cross-contaminated ham during tumbling. However, the pathogen was always absent from the vacuum-packed product provided that heating to a core temperature of 70 degrees C occurred and recontamination during slicing and packing was prevented. The percentage distribution of different species of lactic acid bacteria as well as the uncommon phenotypic characteristics of some strains were discussed.


Atomic layer deposition on gram quantities of multi-walled carbon nanotubes.
Atomic layer deposition (ALD) was employed to grow coaxial thin films of Al(2)O(3) and Al(2)O(3) /W bilayers on multi-walled carbon nanotubes (MWCNTs). Although the MWCNTs have an extremely high surface area, a rotary ALD reactor was successfully employed to perform ALD on gram quantities of MWCNTs. The uncoated and ALD-coated MWCNTs were characterized with transmission electron microscopy and x-ray photoelectron spectroscopy. Al(2)O(3) ALD on untreated MWCNTs was characterized by nucleation difficulties that resulted in the growth of isolated Al(2)O(3) nanospheres on the MWCNT surface. The formation of a physisorbed NO(2) monolayer provided an adhesion layer for the nucleation and growth of Al(2)O(3) ALD films. The NO(2) monolayer facilitated the growth of extremely conformal coaxial Al(2)O(3) ALD coatings on the MWCNTs. Cracks were also observed in the coaxial Al(2)O(3) ALD films on the MWCNTs. After cracking, the coaxial Al(2)O(3) ALD films were observed to slide on the surface of the MWCNTs and expose regions of bare MWCNTs. The Al(2)O(3) ALD film also served as a seed layer for the growth of W ALD on the MWCNTs. The W ALD films can significantly reduce the resistance of the W/Al(2)O(3)/MWCNT wire. The results demonstrate the potential for ALD films to tune the properties of gram quantities of very high surface area MWCNTs.


Confirmatory factor analysis of the Strengths and Difficulties Questionnaire in Singaporean kindergartners.
The Strengths and Difficulties Questionnaire (SDQ) assesses behavioural adjustment in children aged 3 to 16 years. To ascertain the appropriateness of the scale for a specific population, it is important to examine whether the distinctiveness of the scale dimensions can be verified empirically. Confirmatory factor analysis was used to test explicitly which of three models better explain our data, and whether model fit was improved by the addition of method factors. Parents of 411 Singaporean kindergartners completed the SDQ. A four-factor multi-trait multi-method model (Prosocial, Conduct, Hyperactivity, Internalizing and two method factors) provided the best fit to the data. There was strong evidence for convergent and discriminant validity. However, differences in configural loading pattern indicated gender-related differences in the mapping of the SDQ items. Differences in factor structure across countries and gender may reflect differing conceptions of the underlying dimensions, as well as differences in normative expectations. However, our findings may allow its use as a screening tool to identify Singaporean children at risk of emotional and behavioural difficulties.


Multiple sclerosis autoantibodies and antibodies in AIDS may deplete a brain peptide hormone.
Computerized Chou-Fasman analysis of the secondary structure of human T-cell leukemia viruses (HTLV-I, HTLV-II) and human immunodeficiency virus (HIV) envelope proteins revealed that only one antigenic epitope (amino acids EAL) is shared by the three viruses. A similar antigenic epitope is also found in human and rat brain hormone vasopressin-neurophysin. If autoantibodies in multiple sclerosis (MS) are made to the epitope EAL, they may cross-react with the envelope proteins of HTVL. It is speculated that in AIDS patients, antibodies to the antigenic epitope EAL of HIV may cross-react with brain vasopressin-neurophysin, leading to a decline in this brain peptide hormone. Thus it is hypothesized that treatment of both MS and AIDS patients with a synthetic polymer containing the amino acids EAL might eliminate the antibodies to vasopressin-neurophysin and thus alleviate some of the clinical symptoms.


Prevalence of excessive gambling before and after introduction of a national lottery in the United Kingdom: another example of the single distribution theory.
According to the single distribution theory advocated by Rose, the prevalence of a deviant condition such as excessive alcohol consumption depends upon the average level of the corresponding characteristic in the population. The objective of this study was to establish whether the single distribution theory applies to gambling behaviour. Household gambling expenditure in the United Kingdom was examined using Family Expenditure Survey data collected before and after the introduction of a national lottery in November 1994. In cross-sectional analyses, the mean (or median) household expenditure on gambling for each region predicted the prevalence of excessive gambling in that region: the slope of the relationship in 1995-96 was equivalent to an increase of 1.2 (95% CI 0.7-1.7) points in the percentage of households gambling more than 10% of income for every increase of 1 Pound in mean household gambling expenditure. The introduction of the national lottery was associated with an increase in mean household gambling expenditure from 1.45 Pounds to 3.81 Pounds per week, and an increase in the proportion of households gambling more than 10% of total income four-fold from 0.4% to 1.7%. Among households with income of less than 200 Pounds/week, the proportion gambling more than 10% of their income increased from 0.6% to 3.2%. The single distribution theory applies to gambling behaviour. The increase in average gambling expenditure associated with the introduction of a national lottery in the United Kingdom has led to a pronounced increase in the prevalence of excessive gambling, especially in low-income households. This is likely to increase the prevalence of gambling disorders and to exacerbate social inequalities in health.


Comparison of two antacid preparations on intragastric acidity--a two-centre open randomised cross-over placebo-controlled trial.
Rennie and Riopan Gel are 2 of the most well-known and popular over-the-counter antacids; for heartburn symptoms, pain relief is fast with both preparations. A direct comparison with respect to intragastric acidity has not been done yet. The aim of our study was therefore to compare the effects of both preparations on intragastric acidity of fasting volunteers. The study was conducted as an open, randomised, placebo-controlled, 2-centre cross-over study. On different days, 24 healthy adult volunteers (11 males and 13 females) received equimolar acid-neutralising amounts of either Riopan Gel (800 mg magaldrate) or 2 tablets of Rennie (680 mg calcium carbonate and 80 mg magnesium carbonate) or no drug (control) with a wash-out period of at least 4 days between applications. The intragastric pH was measured for 3 h by intragastric pH-metry. The primary endpoint was the median time lag before intragastric pH >3.0 was reached for 10 consecutive min after drug administration. For both antacids, the median pH during the first 30 min after drug administration was statistically significantly different from placebo (p < 0.05), but there was a statistically significant increase in pH during the first 5 min for Riopan Gel only. Compared to placebo, both antacids (Rennie and Riopan Gel) have short-lasting effects on intragastric acidity. There is no statistically significant difference between the 2 preparations, except in the first 5 min, indicating a faster onset of action for Riopan Gel. We conclude that the antacid formulation (tablet or liquid) has little influence on intragastric acidity.


Alternative splicing of the mouse amelogenin primary RNA transcript contributes to amelogenin heterogeneity.
A heterogeneous population of amelogenin proteins is derived from a single copy of the mouse amelogenin gene. To investigate the one gene--multiple protein enigma, we designed a study to distinguish between alternative splicing and proteolytic cleavage models. A pulse of [35S]methionine labeling demonstrated that multiple amelogenins are synthesized concurrently, a result consistent with an alternative splicing mechanism. Using reverse transcription and polymerase chain reaction we cloned a segment from the 5' end of a mouse amelogenin mRNA and connected it to a previously isolated abbreviated cDNA clone. Four additional cDNAs derived from alternatively spliced amelogenin mRNAs have been cloned and characterized. The five transcripts encode amelogenins 180, 156, 141, 74, and 59 amino acids in length.


[Local excision of rectal carcinoma].
From January 1985 till December 1994 109 patients with rectal carcinoma were treated by local excision, in 36 patients a radical operation was performed afterwards. In the assessment of tumor infiltration endosonography was superior to rectal-digital examination. In 34 patients with local excised "low risk" T1-carcinomas and tumor free margins no local recurrence was observed. Two of ten patients with local excised "low risk" T1-carcinoma and no adequate margin of healthy tissue developed a local recurrence. Regarding our results the local excision of "low risk" T1-carcinomas seems justified, if final histological workup reveals an adequate margin of healthy tissue.


Lipidomics: new insight into kidney disease.
Due to the incidence of type-2 diabetes and hypertension, chronic kidney disease (CKD) has emerged as a major public health problem worldwide. CKD results in premature death from accelerated cardiovascular disease and various other complications. Early detection, careful monitoring of renal function, and response to therapeutic intervention are critical for prevention of CKD progression and its complications. Unfortunately, traditional biomarkers of renal function are insufficiently sensitive or specific to detect early stages of disease when therapeutic intervention is most effective. Therefore, more sensitive biomarkers of kidney disease are needed for early diagnosis, monitoring, and effective treatment. CKD results in profound changes in lipid and lipoprotein metabolism that, in turn, contribute to progression of CKD and its cardiovascular complications. Lipids and lipid-derived metabolites play diverse and critically important roles in the structure and function of cells, tissues, and biofluids. Lipidomics is a branch of metabolomics, which encompasses the global study of lipids and their biologic function in health and disease including identification of biomarkers for diagnosis, prognosis, prevention, and therapeutic response for various diseases. This review summarizes recent developments in lipidomics and its application to various kidney diseases including chronic glomerulonephritis, IgA nephropathy, chronic renal failure, renal cell carcinoma, diabetic nephropathy, and acute renal failure in clinical and experimental research. Analytical technologies, data analysis, as well as currently known metabolic biomarkers of kidney diseases are addressed. Future perspectives and potential limitations of lipidomics are discussed.


Rutin exerts antitumor effects on nude mice bearing SW480 tumor.
We previously showed that extracts from Phoradendron serotinum and Croton lechleri exerted in vitro cytotoxic and in vivo antitumor effects and that their main component was rutin (RTN; 3-rhamnosyl-glucosylquercetin). However, it is unknown whether RTN exerts in vivo antitumoral effects on human colon cancer cells. The aim of this work was to evaluate the antitumor effects of RTN on a murine model. Cytotoxic effects of RTN on human cancer and non-tumorigenic cell lines were evaluated using the MTT assay. Different doses of RTN were injected intraperitoneally daily into nu/nu mice bearing tumors of SW480 colon cancer cells during 32 days. The growth and weight of tumors were measured. Serum levels of VEGF, survival time, increase in life span and toxicological effects on body weight and organ weight were also analyzed. RTN showed the highest cytotoxic effects against SW480 cells (IC₅₀ = 125 μM) as compared to the other cancer cells lines and decreased, in a dose dependent manner, the tumor volume and weight of mice bearing SW480 tumor. RTN 20 mg/kg, the highest dose tested, lacked toxic effects on body weight and relative organ weight in mice, increased mean survival time by 50 days, and decreased by 55% the VEGF serum levels compared to untreated mice. RTN exerts in vitro cytotoxic effects on SW480 cells, induces in vivo antitumor effects, lacks toxic effects on mice bearing SW480 tumor and exerts antiangiogenic properties.


Involvement of microRNA-146a in the Inflammatory Response of S tatus Epilepticus Rats.
Status epilepticus (SE), is characterized by high mortality and morbidity, which can cause neuronal injury, neuronal death and alteration of neuronal networks, Recently, inflammation was shown to play a significant role in SE pathogenesis. And miRNA-146a has been shown to be involved in inflammation and to inhibit inflammatory cytokines through NF-κB pathway. In our study, we investigated the relationship between inflammation and miR-146a expression. The SE rat model was induced by lithium-pilocarpine. Hematoxylin and eosin staining (H&E) was performed to observe the histopathology of the rat hippocampus. The expression of COX-2, TNF-α, IL-6 and IL-1β were respectively measured by Western blot and Bio-Plex ProTM Assays. The miR-146a expression in hippocampus tissue was measured by Quantitative real-time PCR. microRNA-146a was highly expressed in the hippocampus of SE rats coupled with increased level of inflammatory cytokines than the normal group. And TQ can attune the expression of inflammatory cytokines, meanwhile, miR-146a was lower in TQ group. The expression of miRNA-146a were positively correlated with the level of inflammatory reaction. TQ may alleviate the inflammatory reaction by inhibiting the NF-κB signaling pathway. Our study shows that miRNA-146a was involved in the inflammatory response and indicated inflammation severity in SE rats. Therefore, miRNA-146a may serve as a potential biomarker or a therapeutic target in SE.


Cerebral vasculitis associated with acute post-streptococcal glomerulonephritis.
The following is a case study involving a 13-year-old girl who presented initial symptoms of an upper respiratory infection. One week later she experienced a short seizure and hours later a grand mal seizure. MRI examination of the brain demonstrated multiple changing abnormal foci of increased density in white and gray matter suggestive of a vasculitic inflammatory pattern. As a result of proteinuria and red cell casts on urinalysis, a renal biopsy was performed resulting in a diagnosis of acute post-streptococcal glomerulonephritis (APSGN). We concluded on the basis of the MRI that vasculitis was secondary to APSGN. The following paper is a description of our findings in this case and a review of the literature supporting this new interpretation of CNS disease due to APSGN.


Evaluation of anterior maxillary alveolar ridge resorption when opposed by the transmandibular implant.
Fifteen edentulous patients with complaints regarding denture comfort and/or function were treated with the transmandibular implant. All patients were restored with conventional maxillary dentures opposed by implant-supported removable prostheses. Two to 4 years after surgery, these patients were evaluated for vertical and horizontal maxillary bone loss with a radiographic analysis developed by the authors. With this technique, attention was focused on vertical alveolar ridge resorption in the anterior maxilla. Although the sample size was small, the findings from this study indicate that vertical bone loss in the anterior maxilla does occur when a maxillary denture is opposed by an implant-supported overdenture. Comparison of these results with a previous study that evaluated anterior maxillary resorption when a complete maxillary denture opposed natural mandibular anterior teeth and a distal extension removable partial denture demonstrated no statistically significant difference.


Expression of EGFR family and steroid hormone receptors in ductal carcinoma in situ of the breast.
The expression of EGFR family and steroid hormone receptors was examined in a series of 40 cases of pure ductal carcinoma in situ (DCIS) of the breast by immunohistochemical staining of paraffin-embedded sections. Hematoxylin and eosin-stained sections were used to classify the tumors according to the published criteria by Holland et al. (Holland R, Peterse JL, Millis RR, et al. Semin Diagn Pathol. 1994;1 1:167-180). Of the tumors 48% were immunoreactive for EGFR, 63% for c-erbB-2, 78% for c-erbB-3, 95% for c-erbB-4, 88% for estrogen receptor (ER) and 80% for progesterone receptor (PR). Statistically significant association between histological grade (differentiation) and c-erbB-2 protein expression was seen (p <.001). In addition, expression of c-erbB-4 protein was associated with c-erbB-2 (p=.004), c-erbB-3 (p=.058), ER (p=.002) and PR (p=.004). It is concluded that c-erbB-2 expression in DCIS is associated with high-grade pathological features, and a higher c-erbB-2 expression is seen in DCIS than in invasive breast carcinomas. A possible association between extensive expression of c-erbB-4 and steroid hormone receptors in proliferative and premalignant breast epithelial cells and the c-erbB-2 expression in DCIS and invasive breast carcinomas is discussed.


Brain serotonin 4 receptor binding is associated with the cortisol awakening response.
Serotonin signalling is considered critical for an appropriate and dynamic adaptation to stress. Previously, we have shown that prefrontal serotonin transporter (SERT) binding is positively associated with the cortisol awakening response (CAR) (Frokjaer et al., 2013), which is an index of hypothalamic-pituitary-adrenal (HPA)-axis output dynamics. Here, we investigated in healthy individuals if cerebral serotonin 4 receptor (5-HT4r) binding, reported to be a proxy for serotonin levels, is associated with CAR. Thirty healthy volunteers (25 males, age range 20-56 years) underwent 5-HT4r PET imaging with [(11)C]-SB207145, genotyping of the SERT-linked polymorphic region (5-HTTLPR), and performed serial home sampling of saliva (5 time points from 0 to 60min from awakening) to assess CAR. The association between 5-HT4r binding in 4 regions of interest (prefrontal cortex, anterior cingulate cortex, pallidostriatum, and hippocampus) and CAR was tested using multiple linear regression with adjustment for age and 5-HTTLPR genotype. Finally, an exploratory voxel-based analysis of the association was performed. CAR was negatively associated with 5-HT4r binding in pallidostriatum (p=0.01), prefrontal cortex (p=0.03), and anterior cingulate cortex (p=0.002), respectively, but showed no association in hippocampus. The results remained significant when taking into account other potentially relevant covariates. In conclusion, our finding reinforces an association between HPA-axis function and serotonin signaling in vivo in humans. We suggest that higher synaptic serotonin concentration, here indexed by lower 5-HT4r binding, supports HPA-axis dynamics, which in healthy volunteers is reflected by a robust CAR.


Autoprocessing of neutrophil elastase near its active site reduces the efficiency of natural and synthetic elastase inhibitors.
An imbalance between neutrophil-derived proteases and extracellular inhibitors is widely regarded as an important pathogenic mechanism for lung injury. Despite intense efforts over the last three decades, attempts to develop small-molecule inhibitors for neutrophil elastase have failed in the clinic. Here we discover an intrinsic self-cleaving property of mouse neutrophil elastase that interferes with the action of elastase inhibitors. We show that conversion of the single-chain (sc) into a two-chain (tc) neutrophil elastase by self-cleavage near its S1 pocket altered substrate activity and impaired both inhibition by endogenous α-1-antitrypsin and synthetic small molecules. Our data indicate that autoconversion of neutrophil elastase decreases the inhibitory efficacy of natural α-1-antitrypsin and small-molecule inhibitors, while retaining its pathological potential in an experimental mouse model. The so-far overlooked occurrence and properties of a naturally occurring tc-form of neutrophil elastase necessitates the redesign of small-molecule inhibitors that target the sc-form as well as the tc-form of neutrophil elastase.