Link between coagulation abnormalities and microcirculatory dysfunction in critically ill patients.
The current review discusses the role of coagulation in microcirculatory abnormalities and whether anticoagulants may improve microvascular perfusion. Microvascular alterations frequently occur in sepsis and ischemia-reperfusion injury. These alterations are due to endothelial dysfunction and interaction of endothelium and circulating cells. Although the activation of coagulation has been extensively shown to occur in these conditions, microthrombosis seems not to be a predominant factor. Nevertheless, the interplay between coagulation, inflammation and the endothelium seems to favor microvascular dysfunction. Several agents with anticoagulant properties, especially activated protein C and antithrombin, improve the diseased microcirculation, but these agents have pleiotropic effects, and it seems unlikely that these beneficial effects are linked to direct inhibition of coagulation. Current evidence does not support the use of pure anticoagulant agents to improve microvascular perfusion. The activation of coagulation may play an indirect role in microvascular dysfunction, through interplay with endothelium and inflammation.


Cardiac Fibroblast GRK2 Deletion Enhances Contractility and Remodeling Following Ischemia/Reperfusion Injury.
G protein-coupled receptor kinase 2 (GRK2) is an important molecule upregulated after myocardial injury and during heart failure. Myocyte-specific GRK2 loss before and after myocardial ischemic injury improves cardiac function and remodeling. The cardiac fibroblast plays an important role in the repair and remodeling events after cardiac ischemia; the importance of GRK2 in these events has not been investigated. The aim of this study is to elucidate the in vivo implications of deleting GRK2 in the cardiac fibroblast after ischemia/reperfusion injury. We demonstrate, using Tamoxifen inducible, fibroblast-specific GRK2 knockout mice, that GRK2 loss confers a protective advantage over control mice after myocardial ischemia/reperfusion injury. Fibroblast GRK2 knockout mice presented with decreased infarct size and preserved cardiac function 24 hours post ischemia/reperfusion as demonstrated by increased ejection fraction (59.1±1.8% versus 48.7±1.2% in controls; P<0.01). GRK2 fibroblast knockout mice also had decreased fibrosis and fibrotic gene expression. Importantly, these protective effects correlated with decreased infiltration of neutrophils to the ischemia site and decreased levels of tumor necrosis factor-α expression and secretion in GRK2 fibroblast knockout mice. These novel data showing the benefits of inhibiting GRK2 in the cardiac fibroblast adds to previously published data showing the advantage of GRK2 ablation and reinforces the therapeutic potential of GRK2 inhibition in the heart after myocardial ischemia.


Influence of vascular endothelial growth factor on bovine corneal endothelial cells in a wound-healing model.
In this study we determined the influence of vascular endothelial growth factor (VEGF) on bovine corneal endothelial cell proliferation and wound healing. Proliferation was determined by measurement of DNA replication as well as by counting of the number of cells present after a defined growth period. In a wound-healing model, reproducible cell-free areas were created within monolayers of cultured bovine corneal endothelial cells and the migration of the cells into these areas was analyzed. The DNA replication and cell proliferation of bovine corneal endothelial cells were not influenced by VEGF. In contrast, in the wound-healing model, VEGF supplementation at concentrations of 1 and 10 ng/ml increased the cell density of the wounded area by 20% and 50%, respectively, as compared with the cell density of wounds left untreated by VEGF. Furthermore, no increase in DNA replication was found in cells involved in wound healing. Our results demonstrate that healing of bovine corneal endothelial cell layers after wounding is predominantly performed by cell migration rather than by proliferation. This migration can be stimulated by the addition of exogenous VEGF.


A novel PCR-based approach to discover miRNA target genes.
MiRNAs are potent regulators of gene expression, and most miRNAs have from several to several thousands of gene targets. Validating the numerous gene targets of a given miRNA remains challenging despite the existence of various tools and databases that predict candidate gene-miRNA pairs. In the present study, we present a high-throughput but flexible method that applies a PCR-based application to simulate the binding of miRNAs to their gene targets. Using hsa-miR-377 as an illustrative example, our method was able to identify 13 potential targets of hsa-miR-377. Moreover, our results include 2 genes (SOD2 and PPM1A) that have already been verified as targets of hsa-miR-377. Our method may provide an alternative way of identifying the gene targets of miRNAs for future research.


Operative strategies in pancreatic trauma.
Pancreatic injuries are relatively uncommon and the choice of an appropriate operative procedure can be difficult. Operations for pancreatic trauma from January 1990 to June 1993 have been reviewed. Fifty-one patients were studied; 13 had blunt trauma, 17 gunshot wounds and 21 stab wounds. The distribution of injuries was: pancreatic head (17), body (15) and tail (19). Most patients had associated injuries of surrounding organs. Operations performed included pancreatoduodenectomy (seven), distal pancreatectomy (seven) and external drainage (35). Five patients (10 per cent) died: two from haemorrhage, one from an acute subdural haematoma and two from multiple organ failure. Ten patients (20 per cent) developed a pancreatic fistula, four following blunt trauma, four after gunshot wounds and two with stab wounds; all but one had undergone drainage procedures. Stab wounds were associated with a low incidence of duct injury and external drainage was usually satisfactory. After blunt trauma and gunshot wounds, duct injuries were common and easily missed; careful exploration by an experienced surgeon is essential. In appropriately selected patients, pancreatic resection can be performed with good result.


Bimodal effect of progesterone on in vitro dopamine function of the rat corpus striatum.
In the present experiment we examined the temporal effects of progesterone (P) upon in vitro dopamine (DA) release from superfused corpus striatum (CS) and medial basal hypothalami (MBH) tissue fragments. In ovariectomized-estrogen-primed adult female rats, P was systemically administered at 0.5, 2, 4, 12 or 24 h prior to sacrifice for CS and at 4 or 24 h for MBH superfusions. Control groups receiving either steroid alone and/or the oil vehicle were included. Progesterone at the 2- to 12-hour periods stimulated spontaneous and amphetamine-evoked DA release and increased post-superfusion tissue concentrations of DA from the CS when compared to controls. At 24 h P apparently produced an active inhibition, with all parameters of DA activity (spontaneous and AMPH-induced DA release), significantly decreased compared to controls. In contrast, P failed to alter the spontaneous or marginal amphetamine response of the MBH but did significantly reduce post-superfusion DA concentration at 24 h. We hypothesize that the dual effect of P upon brain dopaminergic systems (facilitation followed by inhibition), in the CS but not in the MBH represents changes in DA synthesis and release which are coupled in the CS but not in the MBH.


Validation of a model with climatic and flow scenario analysis: case of Lake Burrumbeet in southeastern Australia.
Forecast evaluation is an important topic that addresses the development of reliable hydrological probabilistic forecasts, mainly through the use of climate uncertainties. Often, validation has no place in hydrology for most of the times, despite the parameters of a model are uncertain. Similarly, the structure of the model can be incorrectly chosen. A calibrated and verified dynamic hydrologic water balance spreadsheet model has been used to assess the effect of climate variability on Lake Burrumbeet, southeastern Australia. The lake level has been verified to lake level, lake volume, lake surface area, surface outflow and lake salinity. The current study aims to increase lake level confidence model prediction through historical validation for the year 2008-2013, under different climatic scenario. Based on the observed climatic condition (2008-2013), it fairly matches with a hybridization of scenarios, being the period interval (2008-2013), corresponds to both dry and wet climatic condition. Besides to the hydrologic stresses uncertainty, uncertainty in the calibrated model is among the major drawbacks involved in making scenario simulations. In line with this, the uncertainty in the calibrated model was tested using sensitivity analysis and showed that errors in the model can largely be attributed to erroneous estimates of evaporation and rainfall, and surface inflow to a lesser. The study demonstrates that several climatic scenarios should be analysed, with a combination of extreme climate, stream flow and climate change instead of one assumed climatic sequence, to improve climate variability prediction in the future. Performing such scenario analysis is a valid exercise to comprehend the uncertainty with the model structure and hydrology, in a meaningful way, without missing those, even considered as less probable, ultimately turned to be crucial for decision making and will definitely increase the confidence of model prediction for management of the water resources.


[Angiography in arteriosclerosis].
Advances in angiography of arteriosclerosis have been made possible by improvement of angiographic techniques, materials, such as narrow catheters and gliding guidewires, x-ray apparatus and equipments, including digital subtraction angiography (DSA), computed radiography and magnetic resonance angiography and contrast media. Improved, narrow and yet wide lumen catheters have made it possible to insert the catheter via the axillary or brachial artery. The matrix of DSA was increased to 1024 x 1024 from 512 x 512, and resolution of DSA has been greatly improved. Intravenous DSA has made examination of aneurysm or arterial stenosis or occlusion, easier without much harmful invasion. Ionic or non-ionic low osmolar contrast media has greatly reduced the irritation to the vessels.


A Multinational European Study of Patient Preferences for Novel Diagnostics to Manage Antimicrobial Resistance.
Novel diagnostics are needed to manage antimicrobial resistance (AMR). Patient preferences are important in determining whether diagnostic tests are successful in practice, but there are few data describing the test attributes which matter most to patients. We elicited patients' preferences for attributes of diagnostic tests that could be used to reduce unnecessary antibiotic use in primary care across seven European countries. We used an online stated preference survey, including a discrete choice experiment (DCE). The DCE explored how patients make trade-offs between three key attributes of diagnostic tests: the speed that results were available, confidence in the test results, and how convenient it is to take the test. Individuals were eligible to complete the survey if they had taken antibiotics within the last 2 years and were resident in Germany, Italy, Spain, France, Greece, the Netherlands or the United Kingdom (UK). In total, 988 respondents completed the survey. The DCE responses illustrated that speed was the least important attribute in most countries. Responses from Germany and the Netherlands indicated that confidence was most important in these countries. Responses from the UK, France, Spain and Italy showed convenience as the most important attribute in these countries. Two attributes, confidence and convenience, were jointly favoured by respondents in Greece. Patients in different European countries do not have the same preferences for the attributes of diagnostic tests to manage AMR in primary care. Failure to account for such differences during test development could reduce test uptake, result in continued overuse of antibiotics, and hamper marketisation.


Complete Recovery from Cardiac Arrest Caused by Risperidone-induced Hypothermia.
Hypothermia is a rare adverse effect of risperidone and is associated with ventricular arrhythmias, which may lead to intensive care unit admission and sometimes death. We describe a patient with schizophrenia who was taking risperidone and who, after falling into a hypothermic state, suffered cardiac arrest. Observations of the case and a brief literature review are provided.


[Cytomegalovirus (CMV): virologic protocols for transplantation].
After transplantation, the frequency of reactivation (or superinfection) by CMV is high. Less frequent, but more severe, is the primary form of the disease. The virological procedure to follow up the graft recipients includes: 1) The detection of the infectious virus and/or its antigens in urine, blood, bronchial secretions or brushing, bronchiolar and alveolar washings, ... The laboratory will perform a rapid culture for detection of early CMV antigen (24-72 h) associated with the more classical long-term culture, a reliable but slow method (2-3 weeks). 2) The detection of antibodies, primarily IgM, or even IgA. The usual ELISA for IgG and ELISA modified by addition of an immunocapture on a solid phase are the more reliable techniques. As a rule, the pattern IgM+/IgG- is common for the primary infection. In the case of reactivation (or superinfection), the pattern may be either IgM+/IgG+ or IgM-/IgG+. In the latter case, the increase of the IgG during the next weeks will bring evidence of a reactivation process.


Acquired factor V inhibitor. A problem-based systematic review.
Acquired factor V(FV) inhibitors as a rare bleeding disorder, poses a formidable challenge to treating physicians with limited evidence to guide its management. We systematically reviewed our experience in Singapore and the published literature to determine possible answers to clinical questions formulated on the manifestation and best management of non-bovine thrombin and non-congenital acquired FV inhibitors. The incidence in Singapore was 0.09 cases per million person years (3 cases over 10 years). Seventy-three other cases meeting pre-defined search criteria were found in the published literature. Bleeding occurred in 68.4% of these patients, with mucous membranes being the most common site. Intracranial and retroperitoneal bleeds carried the highest mortality. The mortality rate from bleeding was 12%. There was a tendency for FV levels and PT/aPTT prolongation to predict bleeding but not the inhibitor level. No consistently effective haemostatic agent could be determined, but platelet transfusion should probably be the first line therapy. Among bleeding patients, inhibitors tended to disappear faster with inhibitor elimination therapy (IET) compared to without IET (60 vs. 150 days, p=0.299). IET made no significant difference among non-bleeding patients (p=0.511) and is thus recommended for bleeding patients or those with high bleeding risk. Steroids as single agent IET was effective in the majority of patients. Logical management approaches may be drawn but are limited by small sample size, heterogeneity of reports, and potential publication bias. The inception of a comprehensive registry will provide more reliable data that may verify our findings.


Hybrid E. coli--Mitochondrial ribonuclease P RNAs are catalytically active.
RNase P is a ribonucleoprotein that cleaves tRNA precursors at their 5'-end. Mitochondrion-encoded RNA subunits of mitochondrial RNase P (mtP-RNA) have been identified in jakobid flagellates such as Reclinomonas americana, in the prasinophyte alga Nephroselmis olivacea, and in several ascomycete and zygomycete fungi. While the structures of ascomycete mtP-RNAs are highly reduced, those of jakobids, prasinophytes, and zygomycetes retain most conserved features of their bacterial counterparts. Therefore, these mtP-RNAs might be active in vitro in the absence of a protein subunit, as are bacterial P-RNAs. Here we present a comparative structural analysis including seven newly characterized jakobid mtP-RNAs. We investigate ribozyme activities of mtP-RNAs and find that even the most bacteria-like molecules of jakobids are inactive in vitro. However, when certain domains of jakobid and N. olivacea mtP-RNAs are replaced with those from Escherichia coli, these hybrid RNAs show catalytic activity. In vitro mutagenesis of these hybrid mtP-RNAs shows that various structural elements play a critical role in ribozyme catalysis and provide further support for the presence of these elements in mtP-RNAs. These include GNRA tetraloops in helix P14 and P18 of Jakoba libera, and a remnant P3 pairing in Seculamonas ecuadoriensis. Finally, we will discuss reasons for the failure of mtP-RNAs to show catalytic activity in the absence of P-proteins based on our mutagenesis analysis.


Tree Circumference Changes and Species-Specific Growth Recovery After Extreme Dry Events in a Montane Rainforest in Southern Ecuador.
Under drought conditions, even tropical rainforests might turn from carbon sinks to sources, and tree species composition might be altered by increased mortality. We monitored stem diameter variations of 40 tree individuals with stem diameters above 10 cm belonging to eleven different tree genera and three tree life forms with high-resolution dendrometers from July 2007 to November 2010 and additionally March 2015 to December 2017 in a tropical mountain rainforest in South Ecuador, a biodiversity hotspot with more than 300 different tree species belonging to different functional types. Although our study area receives around 2200 mm of annual rainfall, dry spells occur regularly during so-called "Veranillo del Niño" (VdN) periods in October-November. In climate change scenarios, droughts are expected with higher frequency and intensity as today. We selected dry intervals with a minimum of four consecutive days to examine how different tree species respond to drought stress, raising the question if some species are better adapted to a possible higher frequency and increasing duration of dry periods. We analyzed the averaged species-specific stem shrinkage rates and recovery times during and after dry periods. The two deciduous broadleaved species Cedrela montana and Handroanthus chrysanthus showed the biggest stem shrinkage of up to 2 mm after 10 consecutive dry days. A comparison of daily circumference changes over 600 consecutive days revealed different drought responses between the families concerning the percentage of days with stem shrinkage/increment, ranging from 27.5 to 72.5% for Graffenrieda emarginata to 45-55% for Podocarpus oleifolius under same climate conditions. Moreover, we found great difference of recovery times after longer-lasting (i.e., eight to ten days) VdN drought events between the two evergreen broadleaved species Vismia cavanillesiana and Tapirira guianensis. While Vismia replenished to pre-VdN stem circumference after only 5 days, Tapirira needed 52 days on average to restore its circumference. Hence, a higher frequency of droughts might increase inter-species competition and species-specific mortality and might finally alter the species composition of the ecosystem.


How well will stacked transgenic pest/herbicide resistances delay pests from evolving resistance?
Resistance has evolved to single transgenic traits engineered into crops for arthropod and herbicide resistances, and can be expected to evolve to the more recently introduced pathogen resistances. Combining transgenes against the same target pest is being promoted as the solution to the problem. This solution will work if used pre-emptively, but where resistance has evolved to one member of a stack, resistance should easily evolve for the second gene in most cases. We propose and elaborate criteria that could be used to evaluate the value of stacked traits for pest resistance management. Stacked partners must: target the same pest species; be in a tandem construct to preclude segregation; be synchronously expressed in the same tissues; have similar tissue persistence; target pest species that are still susceptible to at least two stacked partners. Additionally, transgene products must not be degraded in the same manner, and there should be a lack of cross-resistance to stacked transgenes or to their products. With stacked herbicide resistance transgenes, both herbicides must be used and have the same persistence. If these criteria are followed, and integrated with other pest management practices, resistance may be considerably delayed. © 2016 Society of Chemical Industry.


Development and Validation of an ERCC1 Immunohistochemistry Assay for Solid Tumors.
- Excision repair cross-complementation 1 (ERCC1) is a key enzyme in nuclear excision repair pathway and has a critical role in helping remove DNA adducts caused by cross-linking agents, such as platinum-containing cancer chemotherapies and other DNA-damaging therapeutic modalities. ERCC1 expression, evaluated by techniques such as immunohistochemistry, has been associated with clinical response; ERCC1+ tumors are more resistant to cisplatin treatment than are ERCC1- tumors. Although several immunohistochemistry, anti-ERCC1 antibodies are available, the 8F1 clone, in particular, has been used in many studies. Recent evidence has suggested that the 8F1 antibody cross-reacts with at least one other protein, raising concerns about the specificity of this clone. - To design an immunohistochemistry assay to detect ERCC1 levels that show dynamic range and consistent analytic performance. - Two different primary antibodies to ERCC1, clones 4F9 and D6G6, were evaluated on formalin-fixed, paraffin-embedded tissue. We then performed a fit-for-purpose assay validation with the 4F9 clone, which included sensitivity assessment across several solid tumor types and evaluation of analytic parameters, such as precision and reproducibility. - The 4F9 clone was consistently superior to the D6G6 clone in the optimization phase. A range of expression was seen in ovarian, head and neck, non-small cell lung, and esophageal cancer samples when tested with the 4F9 clone. The antibody showed acceptable reproducibility (31.02%) and precision (16.06%). - This assay can be used to assess ERCC1 levels during clinical studies of patient tumors from a variety of tumor types.


Effects of couple drinking events on short-term relationship harmony and discord: An ecological momentary assessment study.
Couples with concordant drinking patterns have higher relationship satisfaction and greater relationship stability over time. Emerging evidence suggests that episodes of drinking with one's intimate partner can have positive immediate consequences for relationship functioning, whereas drinking episodes without partner do not. The present ecological momentary assessment (EMA) study examined the impact of drinking with partner (DWP) and drinking without partner (DWOP) events on immediate and next-day self-reported relationship harmony and discord. Heterosexual, cohabiting community couples with congruent drinking patterns, ages 21-35 (N = 191), made three random reports each day for 30 days. Multilevel modeling was used to examine the impact of DWP and DWOP events on momentary relationship functioning, controlling for quantity of alcohol consumed and for functioning at the previous report. As hypothesized, harmony increased immediately after DWP (but not after DWOP) compared with no drinking for men and women. There were no immediate effects of drinking on discord. There were also positive effects of DWP on next morning harmony, but these were specific to women's drinking reports. Women's DWP and DWOP predicted reduced next morning discord for men; however, men's and women's discord increased as women consumed more drinks the previous evening. Findings suggest that concordant drinking couples may achieve immediate benefits for couple harmony from drinking together. Women's drinking seems to be more impactful than men's on next-day harmony and discord, with greater harmony following women's evening DWP but increased discord associated with heavier evening drinking by women. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Learning from prescribing errors.
The importance of learning from medical error has recently received increasing emphasis. This paper focuses on prescribing errors and argues that, while learning from prescribing errors is a laudable goal, there are currently barriers that can prevent this occurring. Learning from errors can take place on an individual level, at a team level, and across an organisation. Barriers to learning from prescribing errors include the non-discovery of many prescribing errors, lack of feedback to the prescriber when errors are discovered by other healthcare professionals, and a culture that does not encourage reflection on errors together with why they occurred and how they can be prevented. Changes are needed in both systems and culture to provide an environment in which lessons can be learnt from errors and put into practice.


Nonadditivity of faradaic currents and modification of double layer capacitance in the voltammetry of mixtures of ferrocene and ferrocenium salts in ionic liquids.
Electrochemical studies on the Fc + e(-) <==> Fc(+) (Fc = ferrocene) process have been undertaken via the oxidation of Fc and reduction of Fc(+) as the hexafluorophosphate (PF(6)(-)) or tetrafluoroborate (BF(4)(-)) salts and their mixtures in three ionic liquids (ILs) (1-butyl-1-methylpyrrolidinium bis[(trifluoromethyl)sulfonyl]imide, 1-butyl-3-methylimidazolium tetrafluoroborate, and 1-butyl-3-methylimidazolium hexafluorophosphate). Data obtained at macro- and microdisk electrodes using conventional dc and Fourier-transformed large-amplitude ac (FT-ac) voltammetry reveal that diffusion coefficients for Fc and Fc(+) differ significantly and are a function of the Fc and Fc(+) concentration, in contrast to findings in molecular solvents with 0.1 M added supporting electrolyte media. Thus, the faradaic currents associated with the oxidation of Fc (Fc(0/+)) and reduction of FcPF(6) or FcBF(4) (Fc(+/0)) when both Fc and Fc(+) are simultaneously present in the ILs differ from values obtained when individual Fc and Fc(+) solutions are used. The voltammetry for both the Fc(0/+) and Fc(+/0) processes exhibited near-Nernstian behavior at a glassy carbon macrodisk electrode and a platinum microdisk electrode, when each process was studied individually in the ILs. As expected, the reversible formal potentials (E(o)') and diffusion coefficients (D) at 23 +/- 1 degrees C were independent of the electrode material and concentration. However, when Fc and FcPF(6) or FcBF(4) were both present, alterations to the mass transport process occurred and apparent D values calculated for Fc and Fc(+) were found to be about 25-39% and 32-42% larger, respectively, than those determined from individual solutions. The apparent value of the double layer capacitance determined by FT-ac voltammetry from individual and mixed Fc and Fc(+) conditions at the GC electrode was also a function of concentration. Double layer capacitance values increased significantly with the concentration of Fc and FcPF(6) or FcBF(4) when species were studied individually or simultaneously, but had a larger magnitude under conditions where both species were present. Variation in the structure of the ILs and hence mobilities of the ionic species, when Fc and FcPF(6) or FcBF(4) are simultaneously present, is considered to be the origin of the nonadditivity of the faradaic currents and variation in capacitance.


Potential antioxidant, antiinflammatory, and proapoptotic anticancer activities of Kakadu plum and Illawarra plum polyphenolic fractions.
Kakadu plum (Terminalia ferdinandiana Exell, Combretaceae) and Illawarra plum (Podocarpus elatus Endl., Podocarpaceae) extracts were fractionated, using a bioassay-guided approach and screened for antioxidant activity [oxygen radical absorbance capacity (ORAC) and cellular antioxidant activity (CAA) assays] and antiinflammatory activity (nitrite concentration and prostaglandin E(2) release in lipopolysaccharide (LPS)-activated murine macrophages). Among 8 fractions obtained from KP and 5 fractions obtained from IP, fraction KPF5 from KP exhibited superior activity in all assays, with an ORAC value of 3,776 ± 603 μmol Trolox/g DW and a CAA value of 52.2 ± 8.6 μmol quercetin equivalents/g DW. In addition, KPF5 further demonstrated an upregulation of the Nrf2/Keap1 ratio in Hep G2 cells. KPF5 also inhibited the expression of COX-2 and iNOS in LPS-activated murine macrophages, potentially through the NF-κB, p44/42 mitogen activated protein kinase and Akt pathways. KPF5 also induced apoptosis and DNA damage in HT-29 cells, as determined by the cytokinesis block micronucleus cytome assay.


Arterial stiffening with ageing is associated with transforming growth factor-β1-related changes in adventitial collagen: reversal by aerobic exercise.
We tested the hypothesis that carotid artery stiffening with ageing is associated with transforming growth factor-β1 (TGF-β1)-related increases in adventitial collagen and reductions in medial elastin, which would be reversed by voluntary aerobic exercise. Ex vivo carotid artery incremental stiffness was greater in old (29–32 months, n = 11) vs. young (4–7 months, n = 8) cage control B6D2F1 mice (8.84 ± 1.80 vs. 4.54 ± 1.18 AU, P < 0.05), and was associated with selective increases in collagen I and III and TGF-β1 protein expression in the adventitia (P < 0.05), related to an increase in smooth muscle α-actin (SMαA) (myofibroblast phenotype) (P < 0.05). In cultured adventitial fibroblasts, TGF-β1 induced increases in superoxide and collagen I protein (P < 0.05), which were inhibited by Tempol, a superoxide dismutase. Medial elastin was reduced with ageing, accompanied by decreases in the pro-synthetic elastin enzyme, lysyl oxidase, and increases in the elastin-degrading enzyme, matrix metalloproteinase 2. Fibronectin was unchanged with ageing, but there was a small increase in calcification (P < 0.05). Increased incremental stiffness in old mice was completely reversed (3.98 ± 0.34 AU, n = 5) by 10–14 weeks of modest voluntary wheel running (1.13 ± 0.29 km day−1), whereas greater voluntary wheel running (10.62 ± 0.49 km day−1) had no effect on young mice. The amelioration of carotid artery stiffness by wheel running in old mice was associated with reductions in collagen I and III and TGF-β1, partial reversal of the myofibroblast phenotype (reduced SMαA) and reduced calcification (all P < 0.05 vs. old controls), whereas elastin and its modulating enzymes were unaffected. Adventitial TGF-β1-related oxidative stress may play a key role in collagen deposition and large elastic artery stiffening with ageing and the efficacious effects of voluntary aerobic exercise.


Multi-modality of pI distribution in whole proteome.
Multi-modality of pI distribution is a common feature in different whole proteomes. Some researchers considered it relate to the proteins with different subcellular locations, indicating the result of natural selection. We explored the pI distribution of predicted proteomes (including animals, plants, bacterium, archaeans) and random proteome [random protein sequences constructed according to the special amino acid composition and molecular weight (MW) distribution of human predicted proteome]. Our results suggest that the multi-modality is the result of discrete pK(R) values for different amino acids. Amino acid composition and MW distribution of a proteome also contributes to the specific pI distribution. Although protein subcellular location was related to pI value, our analyses revealed that comparing with the random proteome, neither the multi-modality phenomenon nor the distribution bias of pI values is caused by subcellular location. It seems that the multi-modality distribution is just a mathematical fun. The blank region near the neutral pI was caused by the absence of amino acids with neutral pK(R), and suggests that the selection of amino acids with ionizable side chain might be restricted by the requirement for a special pH environment during the origin of life. From this point of view, the special distribution was the result of natural selection.


Remote regional cerebral blood flow consequences of focused infarcts of the medulla, pons and cerebellum.
The aim of this study was to evaluate regional and remote diaschisis of inferior brain stem or cerebellar infarcts in 25 patients presenting with relatively limited lesions. Patients presented with medullary, pontine or cerebellar infarction. Lesions were evaluated on MRI (0.5 T). Regional cerebral blood flow (rCBF) was assessed by means of SPECT, after injection of 9rmTc-hexamethyl propyleneamine oxime (HMPAO) and, when possible, inhalation of 133Xe in the same session. For each method, asymmetry indices (Als), comparing contralateral to ipsilateral rCBF values, were calculated in four areas of each cerebral hemisphere and in the cerebellum and later compared with values obtained in healthy subjects (P = 0.05). Higher rCBF values were observed in the contralateral cerebellum in 2 of 7 patients with selective lateral medullary lesions, and cerebellar Als were significantly increased. When a cerebellar infarct was associated with a lateral medullary lesion, the cerebellar and contralateral hemispheric asymmetries were more severe. Unilateral paramedian pontine infarcts had more frequent consequences on the cerebellum (2 of 3 cases), with rCBF or tracer uptake being reduced in the ipsilateral or the contralateral lobe. Inverse cerebral hemispheric asymmetry could then be observed. Bilateral pontine lesions were difficult to evaluate. Using 99mTc-HMPAO, discrete cerebellar asymmetry was observed in 3 of 6 cases. Pure cerebellar infarcts in the posterior inferior cerebellar artery territory were always associated with a severe ipsilateral flow drop in the cerebellum, and contralateral hemispheric diaschisis was frequent (3 of 4 patients), predominating in the frontotemporal cortex and subcortical structures. This was also more obvious using 99mTC-HMPAO than 133Xe. Variance analysis showed that hemispheric diaschisis was more severe in mixed brain stem and cerebellar infarcts than in pure cerebellar or brain stem lesions. Furthermore, cerebellar and hemispheric AI values were not correlated with measurements of clinical deficits, disability or handicap. Unilateral and limited inferior brain stem lesions can have ipsi- or contralateral consequences on the cerebellum and cerebral hemispheres rCBF. These remote effects are related to lesions of the main pathways joining these structures, resulting in deactivation and, in some cases, overactivation. Contrary to what has been suggested, consequences on cerebral hemispheres are more severe in mixed cerebellar and brain stem infarcts than in pure cerebellar lesions.


Dibromido(2,3-di-2-pyridyl-pyrazine-κN,N)palladium(II).
The Pd(II) ion in the title complex, [PdBr(2)(C(14)H(10)N(4))], is four-coordinated in a slightly distorted square-planar environment by the two pyridine N atoms of the chelating 2,3-di-2-pyridyl-pyrazine (dpp) ligand and two bromide anions. The pyridine rings are considerably inclined to the least-squares plane of the PdBr(2)N(2) unit [maximum deviation = 0.080 (2) Å], making dihedral angles of 64.9 (1) and 66.4 (1)°. The pyrazine ring is perpendicular to the unit plane, with a dihedral angle of 89.0 (1)°. In the crystal, the complex mol-ecules are stacked in columns along the a axis and connected by C-H⋯Br hydrogen bonds, forming a helical chain along the b axis.


Comparison of different degrees of variability in tidal volume to prevent deterioration of respiratory system elastance in experimental acute lung inflammation.
Variable ventilation improves respiratory function, but it is not known whether the amount of variability in tidal volume (VT) can be reduced in recruited lungs without a deterioration of respiratory system elastance. Acute lung inflammation was induced by intratracheal instillation of lipopolysaccharide in 35 Wistar rats. Twenty-eight animals were anaesthetized and ventilated in volume-controlled mode. Lungs were recruited by random variation of VT (mean 6 ml kg(-1), coefficient of variation 30%, normal distribution) for 30 min. Animals were randomly assigned to different amounts of VT variability (n=7 for 90 min per group): 30, 15, 7.5, or 0%. Lung function, diffuse alveolar damage, and gene expression of biological markers associated with cell mechanical stress, inflammation, and fibrogenesis were assessed. Seven animals were not ventilated and served as controls for post-mortem analyses. A VT variability of 30%, but not 15, 7.5, or 0%, prevented deterioration of respiratory system elastance [Mean (SD) -7.5 (8.7%), P<0.05; 21.1 (9.6%), P<0.05; 43.3 (25.9), P<0.05; and 41.2 (16.4), P<0.05, respectively]. Diffuse alveolar damage was lower with a VT variability of 30% than with 0% and without ventilation, because of reduced oedema and haemorrhage. A VT variability of 30, 15, or 7.5% reduced the gene expression of amphiregulin, cytokine-induced neutrophil chemoattractant-1, and tumour necrosis factor α compared with a VT variability of 0%. In this model of acute lung inflammation, a VT variability of 30%, compared with 15 and 7.5%, was necessary to avoid deterioration of respiratory system elastance and was not associated with lung histological damage.


Chemical composition and antioxidant activity of the essential oil of Schisandra chinensis fruits.
This study was designed to examine the composition and in vitro antioxidant activity of the essential oil of Schisandra chinensis fruits. Gas chromatography/mass spectrometry was used to determine the composition of the essential oil. Forty components were identified, representing 90.80% of the oil; ylangene (37.72%), β-himachalene (10.46%) and α-bergamotene (8.57%) were the main components. Antioxidant and radical-scavenging properties were evaluated by means of 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and β-carotene bleaching assay. In the DPPH assay, IC(50) value of the S. chinensis essential oil was determined as 4.17 mg mL(-1). In the β-carotene bleaching assay, the essential oil (1.0 mg mL(-1)) exhibited 25.89% inhibition against linoleic acid oxidation. In both systems, antioxidant capacity of butylated hydroxytoluene was also determined in parallel experiments.


Protein-protein interaction network analysis of cirrhosis liver disease.
Evaluation of biological characteristics of 13 identified proteins of patients with cirrhotic liver disease is the main aim of this research. In clinical usage, liver biopsy remains the gold standard for diagnosis of hepatic fibrosis. Evaluation and confirmation of liver fibrosis stages and severity of chronic diseases require a precise and noninvasive biomarkers. Since the early detection of cirrhosis is a clinical problem, achieving a sensitive, specific and predictive novel method based on biomarkers is an important task. Essential analysis, such as gene ontology (GO) enrichment and protein-protein interactions (PPI) was undergone EXPASy, STRING Database and DAVID Bioinformatics Resources query. Based on GO analysis, most of proteins are located in the endoplasmic reticulum lumen, intracellular organelle lumen, membrane-enclosed lumen, and extracellular region. The relevant molecular functions are actin binding, metal ion binding, cation binding and ion binding. Cell adhesion, biological adhesion, cellular amino acid derivative, metabolic process and homeostatic process are the related processes. Protein-protein interaction network analysis introduced five proteins (fibroblast growth factor receptor 4, tropomyosin 4, tropomyosin 2 (beta), lectin, Lectin galactoside-binding soluble 3 binding protein and apolipoprotein A-I) as hub and bottleneck proteins. Our result indicates that regulation of lipid metabolism and cell survival are important biological processes involved in cirrhosis disease. More investigation of above mentioned proteins will provide a better understanding of cirrhosis disease.


[The overlooked chin trauma in children].
Children seldom experience major facial trauma. However, minor injuries such as facial lacerations, abrasions and dental trauma are more common and naturally get less attention. Children with TMJ trauma usually present with pain, limited jaw movement and even malocclusion. However, these findings are not always recognized and sometimes get overlooked when chin laceration is the only sign. Plain x-rays can contribute to the diagnosis but in many cases are inconclusive and are difficult to interpret. Often, there is a diagnostic dilemma, especially when the clinical findings are not clear and normal occlusion is hard to restore. We present 3 out of 12 children in whom we diagnosed condylar head fracture following minor facial trauma. All children had normal physiological occlusion. Most of them had mild clinical signs such as limited mouth opening, pain and tenderness upon palpation of the joint. Routine plain films were not conclusive. The diagnosis was made or confirmed with CT scan. Early diagnosis of TMJ fracture in children is mandatory for the prevention of long term functional and esthetically debilitating sequela. A thorough clinical examination and normal occlusion do not rule out condylar head fracture after chin trauma. Plain films are of limited value in the diagnosis of such fractures. CT should be considered for the evaluation of these children.


Unilateral digital clubbing in hemiplegia due to a putaminal hemorrhage.
Digital clubbing usually occurs bilaterally; however, unilateral clubbing can be seen in hemiplegia and local vascular lesions. We highlight a case of unilateral digital clubbing due to putaminal hemorrhage. Further accumulation of cases will enable exploration of the mechanisms of clubbing.


Mass production of nitrogen and oxygen codoped carbon nanotubes by a delicately-designed Pechini method for supercapacitors and electrocatalysis.
Heteroatom-doped carbon nanotubes (CNTs) have great potential in various fields owing to their extraordinary electronic, structural, and mechanical properties. However, large-scale production of heteroatom-doped CNTs in a simple, economical, and highly efficient manner still remains challenging. Here, we report a modified Pechini method (MPM) for high-yield synthesis of N- and O-codoped CNTs (N,O-CNTs), by rapid pyrolysis of a NiCo-polymer precursor forming via a simple sol-gel process. The carbon source (i.e., citric acid) is inexpensive, and the NiCo-polymer material is the single precursor for the preparation of N,O-CNTs via a thermolysis process without the introduction of additional catalysts or carrier gas. Appropriate NiCo-organic coordination and controlled pyrolysis (i.e. heating rate, pyrolysis temperature, and holding time) are demonstrated to play vital roles in this MPM, which are critical for quick generation of small NiCo nanocatalysts with high catalytic activity and simultaneous formation of sufficient space inside the material. The growth mechanism is well studied. Benefitting from the hierarchically porous structure and the synergistic effect of N,O-codoping in the CNTs, the as-synthesized N,O-CNTs manifest excellent electrochemical performance in both supercapacitors and electrocatalysis. Density functional theory simulations show that N and O dopants could increase the densities of states of CNTs near the Fermi level and charge densities of adjacent C atoms, thus leading to improved electrochemical activity. We anticipate that this work will open up a new avenue for a high-yield and economical synthesis of heteroatom-doped CNTs for energy-related applications and beyond.


[Dissociation of visual evoked responses to hemi-field or full-field flash-checkerboard stimulation].
Unexpected visual evoked responses (VERs) were recorded in 5 subjects with tumoral, ischemic or hemorrhagic lesions of the retrochiasmatic visual pathways. The flash pattern stimulation was always binocular and involved full-field and half-field stimuli. In these 5 cases, the total field VER was asymmetrical with anomalies on the affected occipital region. However half-field VERs P100 contralateral to the stimulus were noted both on the normal and on the affected occipital region. One can ask if this is not an electrophysiological equivalent of the clinical relative hemianopsia.


Value of CEA level determination in gallbladder bile in the diagnosis of liver metastases secondary to colorectal adenocarcinoma.
The relevance of colorectal adenocarcinoma lies in its high incidence, with the liver being the organ most frequently affected by distant metastases. Liver metastases occur in 40 to 50% of patients with colorectal adenocarcinoma, accounting for approximately 80% of deaths in the first three postoperative years. Nevertheless, despite this, they are occasionally susceptible to curative treatment. The present investigation focused on the relationship between the level of carcinoembryonic antigen (CEA) in gallbladder bile and the presence of liver metastases secondary to colorectal adenocarcinoma. Diagnostic test study. Surgical Gastroenterology Discipline at the São Paulo Hospital, São Paulo, Brazil. Forty-five patients with colorectal adenocarcinoma were studied, 30 without liver metastases (group I), and 15 with liver metastases (group II). Diagnosis of liver metastases was made through computed tomography, magnetic resonance imaging and computed tomography during arterial portography. Samples of peripheral blood, portal system blood, and gallbladder bile were collected from patients during the surgical procedure. A control group composed of 18 organ donors underwent the same material collection procedures. CEA level determination was made through fluoroimmunoassay. Mean CEA value in peripheral serum was 2.0 ng/ml (range: 0.7 to 3.8 ng/ml) in the control group, 11.4 ng/ml (range: 0.5 to 110.3 ng/ml) in group I, and 66.0 ng/ml (range: 2.1 to 670 ng/ml) in group II. In the portal system, serum mean values found were 1.9 ng/ml (range: 0.4 to 5.0 ng/ml) in the control group, 15.3 ng/ml (range: 0.8 to 133.3 ng/ml) in group I, and 70.8 ng/ml (range: 1.8 to 725 ng/ml) in group II. Mean values found in gallbladder bile were 4.1 ng/ml (range: 1.0 to 8.6 ng/ml) in the control group, 14.3 ng/ml (range: zero to 93.0 ng/ml) in group I, and 154.8 ng/ml (range: 14.0 to 534.7 ng/ml) in group II. The CEA level in gallbladder bile is elevated in patients with liver metastases. Determination of CEA both in peripheral serum and in gallbladder bile enabled patients with liver metastases to be distinguished from those without such lesions. The level of CEA in gallbladder bile, however, seems to lead to a more accurate diagnosis of liver metastases secondary to colorectal adenocarcinoma.


One-year objective and functional outcomes of a randomized clinical trial of vaginal mesh for prolapse.
The purpose of this study was to show 12-month outcomes of a randomized trial that compared vaginal prolapse repair with and without mesh. Women with stage ≥2 prolapse were assigned randomly to vaginal repair with or without mesh. The primary outcome was prolapse stage ≤1 at 12 months. Secondary outcomes included quality of life and complications. All 65 evaluable participants were followed for 12 months after trial stoppage for mesh exposures. Thirty-two women had mesh repair; 33 women had traditional repair. At 12 months, both groups had improvement of pelvic organ prolapse-quantification test points to similar recurrence rates. The quality of life improved and did not differ between groups: 96.2% mesh vs 90.9% no-mesh subjects reported a cure of bulge symptoms; 15.6% had mesh exposures, and reoperation rates were higher with mesh. Objective and subjective improvement is seen after vaginal prolapse repair with or without mesh. However, mesh resulted in a higher reoperation rate and did not improve 1-year cure.


DPP-4 Inhibitor Reduces Central Blood Pressure in a Diabetic and Hypertensive Patient: A Case Report.
Hypertension and type 2 diabetes mellitus (DM) are among the main risk factors for the development of cardiovascular disease. Pharmacotherapy for DM should not only improve blood glucose control, but also provide beneficial glucose-independent cardiovascular effects. The central systolic blood pressure (SBP) has become more important than the brachial SBP in the assessment of cardiovascular risk.This case report describes the effect of vildagliptin, a dipeptidyl peptidase-4 (DPP-4) inhibitor, on the central SBP in a 54-year-old woman with hypertension and DM. She was submitted to applanation tonometry (AT) before and after vildagliptin association. AT of the radial artery is a non-invasive method that indirectly assesses arterial stiffness by calculating the central SBP and the augmentation index (AIx).After 3 months of follow-up using vildagliptin, central SBP and AIx were improved. Moreover, she presented better glycemic control.This case suggests an effect of DPP-4 inhibitor on arterial stiffness parameter (central SBP) in a hypertensive and diabetic patient, which shows a glucose-independent beneficial cardiovascular effect of this group of drugs.


[Design of high-efficiency double compound parabolic concentrator system in near infrared noninvasive biochemical analysis].
High signal-to-noise ratio (SNR) of system is necessary to obtain accurate blood components in near infrared noninvasive biochemical analysis. In order to improve SNR of analytical system, high-efficiency double compound parabolic concentrator (DCPC) system was researched, which was aimed at increasing light utilization efficiency. Firstly, with the request of collection efficiency in near infrared noninvasive biochemical analysis, the characteristic of emergent rays through compound parabolic concentrator (CPC) was analyzed. Then the maximum focusing angle range of the first stage CPC was determined. Secondly, the light utilization efficiency of truncated type was compared with standard DCPC, thus the best structure parameters of DCPC system were optimized. Lastly, combined with optical parameters of skin tissue, calculations were operated when incident wavelength is 1 000 nm. The light utilization efficiency of DCPC system, CPC-focusing mirror system, and non-optical collecting system was calculated. The results show that the light utilization efficiency of the three optical systems is 1.46%, 0.84% and 0.26% respectively. So DCPC system enhances collecting ability for human diffuse reflection light, and helps improve SNR of noninvasive biochemical analysis system and overall analysis accuracy effectively.


Identification of contaminant coatings over rough surfaces using polarized infrared scattering.
An optical technique to identify the presence of chemical coatings over rough surfaces is described. It is based on the selective use of elements of the 4 x 4 Mueller matrix. The full-wave theory of electromagnetic scattering is used to predict six independent Mueller elements from randomly rough uncoated (dry) and coated (wet) surface materials as functions of the media complex dielectric coefficients, backscattering angle, and midinfrared wavelengths of laser-beam excitations that are polarization modulated. The set of independent elements at beam wavelengths and backscattering angles [M(mn) (lambda(i), lambda(i))] most sensitive to i optically thick contaminant coatings are statistically obtained from the full-wave database, and detection-parameter sets [lambda(i), lambda(i)] are inputs to another algorithm designed to identify the contaminant coating (when present and interacted by the irradiating beams). These algorithms facilitate the operation of a multi-CO(2) laser-ellipsometer facility now under development at the U.S. Army Chemical Research, Development, and Engineering Center for the remote detection of chemical or biological surface contaminants.


Equol: A Bacterial Metabolite from The Daidzein Isoflavone and Its Presumed Beneficial Health Effects.
Epidemiological data suggest that regular intake of isoflavones from soy reduces the incidence of estrogen-dependent and aging-associated disorders, such as menopause symptoms in women, osteoporosis, cardiovascular diseases and cancer. Equol, produced from daidzein, is the isoflavone-derived metabolite with the greatest estrogenic and antioxidant activity. Consequently, equol has been endorsed as having many beneficial effects on human health. The conversion of daidzein into equol takes place in the intestine via the action of reductase enzymes belonging to incompletely characterized members of the gut microbiota. While all animal species analyzed so far produce equol, only between one third and one half of human subjects (depending on the community) are able to do so, ostensibly those that harbor equol-producing microbes. Conceivably, these subjects might be the only ones who can fully benefit from soy or isoflavone consumption. This review summarizes current knowledge on the microorganisms involved in, the genetic background to, and the biochemical pathways of, equol biosynthesis. It also outlines the results of recent clinical trials and meta-analyses on the effects of equol on different areas of human health and discusses briefly its presumptive mode of action.


Autoxidation of lipids and antioxidation by alpha-tocopherol and ubiquinol in homogeneous solution and in aqueous dispersions of lipids: unrecognized consequences of lipid particle size as exemplified by oxidation of human low density lipoprotein.
Recent studies on the initial stages in oxidation of low density lipoprotein (LDL) have revealed certain previously unrecognized similarities to emulsion polymerization and some quite unexpected features including the following: (i) ascorbate is an extremely effective antioxidant for LDL containing alpha-tocopherol (alpha-TOH); (ii) in the presence of alpha-TOH and in the absence of both ascorbate and ubiquinol 10 (Q10H2), oxidation of LDL occurs via a free radical chain; (iii) Q10H2 is a much better antioxidant for LDL than alpha-TOH, although the reverse is true in homogeneous systems. We show here that these problems can be solved on the basis of three simple hypothesis, each of which is based on known chemistry: (i) alpha-TOH in LDL can be regenerated from its radical, alpha-TO., by ascorbate; (ii) in the absence of ascorbate and Q10H2, the alpha-TOH in LDL acts as a chain-transfer agent rather than as a radical trap; (iii) Q10H2 is a much more effective chain-breaking antioxidant than alpha-TOH in LDL because the semiquinone radical Q10H. exports its radical character from the LDL into the aqueous phase. Our conclusions imply that the search for better antiatherosclerotic drugs might profitably focus on antioxidants capable of exporting radicals from LDL particles or otherwise increasing the traffic of radicals between particles.


Update: Investigations of West Nile virus infections in recipients of organ transplantation and blood transfusion.
An investigation involving CDC, the Food and Drug Administration (FDA), the Health Resources and Services Administration (HRSA), the Georgia Division of Public Health, and the Florida Department of Health identified West Nile virus (WNV)-associated illnesses in four recipients of organs from the same donor. Although the transplanted organs were the source of infection for the four organ recipients, the source of the organ donor's infection remains unknown; an investigation of the numerous transfusions received by the organ donor is ongoing.


[Minimal invasive elastic intramedullary nails and external fixation for treatment of comminuted closed fracture of tibia-fibula shaft].
To investigate the clinical results of external fixation and AO titanium elastic intramedullary nailing for treatment of tibia-fibula comminuted closed fractures. From June 2010 to June 2012,58 patients with tibia-fibula comminuted closed fractures were treated with external fixation and AO titanium elastic intramedullary nailing, including 31 males and 27 females with an average age of 38.5 years old ranging from 21 to 57 years old. According to the system of AO Classification, the fractures were classified as type B1 in 9 cases,type B2 in 7 cases, type B3 in 10 cases, type Cl in 14 cases, type C2 in 12 cases,and type C3 in 6 cases. According to the system of Winquist-Hanson,the fractures' comminuted were classified as grade 1 in 23 cases, grade 2 in 17 cases, grade 3 in 12 cases, and grade 4 in 6 cases. According to the system of Johner-Wruhs, clinical results were compared between different type and grade groups by the time of last followed-up. All 58 patients were followed up with an average time of 6.8 months (ranged from 18 to 36 weeks). All fractures had clinical healing with an average time of 28 weeks (ranged from 24 to 32 weeks). The total rate of good to excellent results was 91.4%. The rate of good to excellent in the group of grade 1 was higher than that of other grades. The complication rates and fracture healing time would increase respectively with higher Winquist-Hanson's grade. The complication rates in the group of type C3 was higher than that of other types, but the rate of good to excellent was lower than that of other types. The complication rates in the group of type B1 was lower than that of other types,but the rate of good to excellent was higher than that of other types. Minimal invasiveusing AO titanium elastic intramedullary nailing combined with external fixation for treatment of tibia-fibula fractures especially for the multiple-segment,long spiral mild-to-moderate comminuted with hidden fracture can get satisfactory reduction and reliable fixation,it conformes to the principle of BO completely, protects the fracture end blood supply,reduces the external fixation time, has less skin soft tissue complications, postoperative function recovered satisfactorily.


[Optimization of mobilization regimes of blood hemopoietic stem cells in patients with multiple myeloma].
To determine an optimal cyclophosphamide dose in the mobilization scheme providing adequate collection of CD34+ cells in patients with multiple myeloma (MM), to optimize the time of initiation of granulocytic colony-stimulating factor (G-CSF) administration, to study effects of induction therapy schemes on results of mobilization and collection of CD34+ cells. Department of hemoblastoses chemotherapy and bone marrow transplantation of the Russian Hematological Center performed mobilization of autologous blood hemopoietic stem cells (BHSC) in 93 MM patients treated in 2001-2010. This was done with cyclophosphamide and G-CSF. The former was used in 59 cases in a dose 6 g/m2, in 34 cases - 4 g/m2. Myelotoxic agranulocytosis after cyclophosphamide administration developed in all the patients and was observed for 3-10 days (median 5 days). Agranulocytosis ran without documented infections in 51 (54.8%) patients, with febril fever - in 42 (45.2%) patients. Cepticemia, pneumonia, necrotic enteropathy, stomatitis, herpetic lesion of the skin were registered in 9, 4, 11, 14 and 6 cases, respectively. Severe thrombocytopenia (< 30 x 10(9)/l) occurred more frequently in administration of 6 g/m2 cyclophosphamide. It was corrected with 2-5 transfusions of thromboconcentrates, only 1 transfusion was needed after the dose 4 g/m2. Collection of CD34+ cells started in leukocyte level over 3.5 x 10(9)/l on mobilization day 12-20 (median day 15). The day of the first leukocytapheresis did not depend on the day of the first introduction of G-CSF. Duration of G-CSF administration was significantly shorter in the start of its use after leukocyte count decrease under 1.0 x 10(9)/l. Conduction of 1 to S (median 2) leukocytapheresis was needed for collection of BHSC. Sufficient for 2 autotransplantations number of BHSC were stored in 90 of 93 patients. Cyclophosphamide administration in a dose 6 g/m2 allowed collection of cells sufficient for one autotransplantation for the first leukapheresis in 52 (88.1) patients. A total number of CD34+ cells over 4 x 10(6) cells/kg were collected in 56 (94.9%) patients. In administration of cyclophosphamide in a dose 4 g/m2 mobilization was effective in all 34 patients. The first leukapheresis provided sufficient for one autotransplantation number of cells in 29 (85.3%) patients. Administration of high cyclophosphamide doses in combination with G-CSF is an effective and safe method of BHSC mobilization providing collection of adequate number of CD34+ cells for double autotransplantation in 96.8% patients. Cost effective is the start of G-CSF administration in the fall of leukocytes under 1.0 x 10(9)/l. Cyclophosphamide dose 4 g/m2 provides collection of CD34+ cells number sufficient for two autotransplantations in moderate thrombocytopenia and in less number of substitute transfusions in the absence of serious toxic complications.


UpSET-ting the balance: modulating open chromatin features in metazoan genomes.
Appropriate gene expression relies on the sophisticated interplay between genetic and epigenetic factors. Histone acetylation and an open chromatin configuration are key features of transcribed regions and are mainly present around active promoters. Our recent identification of the SET-domain containing protein UpSET established a new functional link between the modulation of open chromatin features and active recruitment of well-known co-repressors in metazoans. Structurally, the SET domain of UpSET resembles H3K4 and H3K36 methyltransferases; however, it is does not confer histone methyltransferase activity. Rather than methylating histones to regulate gene expression like other SET domain-containing proteins, UpSET fine-tunes transcription by modulating the chromatin structure around active promoters resulting in suppression of expression of off-target genes or nearby repetitive elements. Chromatin modulation by UpSET occurs in part through its interaction with histone deacetylases. Here, we discuss the different scenarios in which UpSET could play key roles in modulating gene expression.


A Numerical Simulation of Cell Separation by Simplified Asymmetric Pinched Flow Fractionation.
As a typical microfluidic cell sorting technique, the size-dependent cell sorting has attracted much interest in recent years. In this paper, a size-dependent cell sorting scheme is presented based on a controllable asymmetric pinched flow by employing an immersed boundary-lattice Boltzmann method (IB-LBM). The geometry of channels consists of 2 upstream branches, 1 transitional channel, and 4 downstream branches (D-branches). Simulations are conducted by varying inlet flow ratio, the cell size, and the ratio of flux of outlet 4 to the total flux. It is found that, after being randomly released in one upstream branch, the cells are aligned in a line close to one sidewall of the transitional channel due to the hydrodynamic forces of the asymmetric pinched flow. Cells with different sizes can be fed into different downstream D-branches just by regulating the flux of one D-branch. A principle governing D-branch choice of a cell is obtained, with which a series of numerical cases are performed to sort the cell mixture involving two, three, or four classes of diameters. Results show that, for each case, an adaptive regulating flux can be determined to sort the cell mixture effectively.


Acute intravenous injection toxicity study of MIBG in mice.
Metaiodobenzylguanidine (MIBG) is an analog of norepinephrine. I-131-labeled MIBG has been thought to be safe and effective in the evaluation of neuroendocrine tumors, mainly in neuroblastoma and pheochromocytoma. This article describes the acute toxicity of MIBG in imprinting control region (ICR) mice. Treated mice were administered with MIBG at dose levels of 75, 150, and 300ng/kg with dose volumes of 20mL/kg. The control mice were administered 20mL/kg of vehicle control. The mice were observed for 14 days. Observations included general demeanor, clinical signs, mortality, body weights/total body-weight gains, and gross necropsy findings. None of the animals died during the 14-day study period. There was no difference in body weights among all treated and control mice.


[Genetic variation in the apolipoprotein B gene].
It is well known that coronary heart disease (CHD) is multifactorial, with environmental and inherited risk factors both playing a role. Apolipoprotein B (apo B) is of major importance in lipoprotein metabolism and might play a central role in atherogenesis. The apo B gene is the obvious candidate gene to study the relations between lipid concentrations and CHD. Some rare mutations in the apo B gene affect plasma cholesterol levels, leading to either familial hypobetalipoproteinemia or familial defective apolipoprotein B100. Other frequent polymorphisms have little biological effect but, because of their high frequency, might contribute to the development of CHD in a given population. Many apo B gene polymorphisms are associated with variations in plasma lipid concentrations, including the response of plasma lipids to dietary intervention, and peripheral and coronary atherosclerosis. Age, body mass index and gender affect the degree and nature of the association between apo B genetic markers and normal lipid and lipoprotein levels. However, negative and contradictory results have also been reported. One likely explanation is differences between studies, including populations of different geographic origin, arbitrary definition of cases and controls and multiple criteria for CHD. Future work on the effect of the apo B locus on hyperlipidaemia and atherosclerosis must involve large numbers of patients belonging to carefully defined populations. Prospective studies using a combination of genetic markers in well-defined populations should lead to firm conclusions on the role of apo B in atherogenesis and coronary heart disease.


Antibody-mediated enhancement of Legionella pneumophila-induced interleukin 1 activity.
The ability of antibody specific for Legionella pneumophila to enhance the induction of interleukin 1 (IL-1) production by murine peritoneal, splenic, and pulmonary macrophages in response to the bacterium was examined. Two preparations of L. pneumophila were utilized, a formalin-killed whole-cell preparation and viable bacteria. We measured both secreted (sIL-1) and cell membrane-associated (mIL-1) activities after incubation of the macrophages with the bacterial preparations in the presence or absence of the antibody. Both bacterial preparations induced sIL-1 and mIL-1 activities in each of the macrophage populations tested. These activities were generally enhanced by pretreating the bacteria with antibody, with the greatest enhancing activity observed for the formalin-killed preparations at lower doses of bacteria.


Bio-ecological approach to cognitive assessment.
Because determining a child's educational program usually takes into consideration the child's cognitive functioning, it is vital that those who are involved in this enterprise recognize that many variables contribute to and explain performance. Thus, careful consideration, assessment, and planning are mandatory to plan a child's individual educational program. This article emphasizes that cognitive assessment is both a formal and an informal process that occurs in several contexts-the school, the home, and the community. Therefore, when psychological evaluators assess the cognitive performance of a child, it is necessary to analyze the complete of his or her social/emotional environments that have contributed to his/her current cognitive performance levels. Thus, it is vital to examine the orientation of family members, the academic assistance students receive at home, as well as the current and previous classroom environments in which the student has been educated. Six nonpsychometric measures (using item equivalency, test-teach-retest, and contextualization) were developed by the authors and are presented in recognition that there are many factors that one must consider when interpreting the performance of a child.


HMB-45 reactivity in adrenal pheochromocytomas.
Melanoma-specific antibody (HMB-45) is highly specific for junctional nevi, malignant melanomas, and related lesions. Rarely have other benign or neoplastic tissues demonstrated positivity with the monoclonal antibody following purification. In the present study, four of 12 pheochromocytomas contained chief cells that reacted with HMB-45. We discuss the clinical and pathologic implications of this finding.


Born Into Addiction: Neonatal Abstinence Syndrome and Pediatric Dentistry.
Drug overdose and opioid-involved deaths continue to increase across the United States. An end-condition related to opioid abuse is neonatal abstinence syndrome (NAS). The implications for pregnant or post-partum women using an opioid, whether prescribed or illicit, can have significant effects on their infant. First described in the 1970s, NAS is defined as a postnatal drug withdrawal syndrome that primarily occurs in opioid-exposed infants. There are no nationally accepted, evidence-based treatment protocols for these infants. Pediatric dentists should be cognizant of potential dental development issues and developmental delays in children with NAS. Basic behavior guidance techniques, such as tell-show-do, may not be as effective. In addition, adherence to prescription drug monitoring programs can help identify patients who may have received multiple opioids, alerting the provider to possible misuse or abuse. The purpose of this paper was to provide background information on the incidences and potential dental implications for children with neonatal abstinence syndrome.


Similarities and differences between non-radiographic and radiographic axial spondyloarthritis: The patient perspective from the Spanish atlas.
Although non-radiographic axial spondyloarthritis (EspAax-nr) is well understood within health institutions, being considered along with radiographic EspAax (EspAax-r) as part of the same disease spectrum, patient understanding is unknown. The aim is to describe the patient's knowledge of the EspAax-nr entity. Atlas 2017, promoted by the Spanish Federation of Spondylarthritis Associations (CEADE), aims to comprehensively understand the reality of EspAax patients from a holistic approach. A cross-sectional on-line survey of unselected patients with self-reported EspAax diagnosis from Spain was conducted. Participants were asked to report their diagnosis. Socio-demographic, disease characteristics and patient-reported outcomes (PROs) were compared between those patients self-reporting as EspAax-nr and EspAax-r. 634 EspAax patients participated. Mean age 45.7±10.9 years, 50.9% female and 36.1% university-educated. 35 (5.2%) self-reported as EspAax-nr. Compared to EspAax-r patients, those with EspAax-nr were more frequently women (48.6% vs 91.4%, p<0.001), had longer diagnostic delay (10.1±8.9 vs 8.5±7.6 years), higher psychological distress (GHQ-12: 7.5±4.9 vs 5.6±4.4) and similar degree of disease activity (BASDAI: 5.7±2.1 vs 5.7±2.0), and unemployment rates (20.0% vs 21.6%). 20.0% of EspAax-nr received biologics vs 36.9% of EspAax-r, p=0.043. Visits to the rheumatologist in the past year were similar in both groups (3.8±4.5 vs 3.2±3.8), while GP visits were much higher within EspAax-nr (8.0±10.7 vs 4.9±13.3 p=0.003). For the first time, EspAax-nr characteristics and PROs have been analyzed from the patient's perspective. Both groups reported similar trends with the exception of EspAax-nr being more frequently women, younger, having longer diagnostic delay and lower use of biologic therapy.


Platelet-activating factor and Escherichia coliO157:H7 infections.
The role of platelet-activating factor (PAF), a phospholipid inflammatory mediator, in Escherichia coli O157:H7-associated hemolytic uremic syndrome (HUS) is unknown. PAF is synthesized by diverse cells and is degraded by PAF-acetylhydrolase (PAF-AH). Deficient PAF-AH activity results from a G-->T transversion at position 994 of exon 9. We examined children infected with E. coliO157:H7 to determine if PAF levels or the PAF-AH ( G994T) mutation reflects the risk of developing HUS. Plasma PAF concentrations were determined using chloroform/methanol extraction, thin layer chromatography purification, and scintillation proximity assay in 10 patients with uncomplicated infection (UI), 10 infected patients who subsequently developed HUS (pre-HUS), 5 HUS patients, and 8 healthy controls. The PAF-AH ( G994T) allele frequency was determined in 52 UI children, 15 with HUS, and 11 controls. Wilcoxon rank sum tests were performed to test differences in location (median) of pairs of groups. PAF levels were higher in the UI ( P=0.04) and pre-HUS ( P=0.01) groups than in healthy controls. No subject had the PAF-AH ( G994T) allele. Thus, elevated plasma PAF levels occur in E. coliO157:H7-infected children, even without HUS, but diminish when HUS develops. The PAF-AH ( G994T) allele does not contribute to the risk of developing HUS.


Radiographic appearance of confirmed pulmonary lymphoma in cats and dogs.
Herein we describe the thoracic radiographic appearance of confirmed pulmonary lymphoma. Patients with thoracic radiographs and cytologically or histologically confirmed pulmonary lymphoma were sought by contacting American College of Veterinary Radiology members. Seven cats and 16 dogs met the inclusion criteria, ranging in age from 4 to 15 years. Method of diagnosis was via ultrasound-guided cytology (four), surgical biopsy (two), ultrasound-guided biopsy (one), and necropsy (16). Radiographic findings varied but ranged from normal (one) to alveolar (six) and/or unstructured interstitial infiltrates (11), nodules and/or masses (eight), and bronchial infiltrates (four). Additional thoracic radiographic findings included pleural effusion and lymphadenopathy. The results of this evaluation indicate a wide variability in thoracic radiographic abnormalities in cats and dogs with pulmonary lymphoma.


Preparation of a primary argon beam for the CERN fixed target physics.
The fixed target experiment NA61 in the North Area of the Super Proton Synchrotron is studying phase transitions in strongly interacting matter. Up to now they used the primary beams available from the CERN accelerator complex (protons and lead ions) or fragmented beams created from the primary lead ion beam. To explore a wider range of energies and densities a request was made to provide primary argon and xenon beams. This paper describes the results of the setting up and 10 week test run of the Ar(11+) beam from the 14.5 GHz ECR ion source and the linear accelerator (Linac3) at CERN.


Thyroid function tests in patients taking thyroid medication in Germany: Results from the population-based Study of Health in Pomerania (SHIP).
Studies from iodine-sufficient areas have shown that a high proportion of patients taking medication for thyroid diseases have thyroid stimulating hormone (TSH) levels outside the reference range. Next to patient compliance, inadequate dosing adjustment resulting in under- and over-treatment of thyroid disease is a major cause of poor therapy outcomes. Using thyroid function tests, we aim to measure the proportions of subjects, who are under- or over-treated with thyroid medication in a previously iodine-deficient area. Data from 266 subjects participating in the population-based Study of Health in Pomerania (SHIP) were analysed. All subjects were taking thyroid medication. Serum TSH levels were measured using immunochemiluminescent procedures. TSH levels of < 0.27 or > 2.15 mIU/L in subjects younger than 50 years and < 0.19 or > 2.09 mIU/L in subjects 50 years and older, were defined as decreased or elevated, according to the established reference range for the specific study area. Our analysis revealed that 56 of 190 (29.5%) subjects treated with thyroxine had TSH levels outside the reference range (10.0% elevated, 19.5% decreased). Of the 31 subjects taking antithyroid drugs, 12 (38.7%) had TSH levels outside the reference range (9.7% elevated, 29.0% decreased). These proportions were lower in the 45 subjects receiving iodine supplementation (2.2% elevated, 8.9% decreased). Among the 3,974 SHIP participants not taking thyroid medication, TSH levels outside the reference range (2.8% elevated, 5.9% decreased) were less frequent. In concordance with previous studies in iodine-sufficient areas, our results indicate that a considerable number of patients taking thyroid medication are either under- or over-treated. Improved monitoring of these patients' TSH levels, compared to the local reference range, is recommended.


Using a computer to manage the radiation safety and quality control activities in diagnostic radiology.
Software has been developed to aid in the management of a radiation safety quality control program in diagnostic radiology. The core of the system is a data base of radiation safety activities. Software design goals were the prioritization, scheduling, and reporting of these activities. Computerization has helped organize the timely performance of surveys necessary for compliance with regulatory and accreditation agencies. Clear, concise program documentation and reporting are also provided.


Automatic quantification of ejection fraction from gated myocardial perfusion SPECT.
We have developed a completely automatic algorithm to quantitatively measure left ventricular ejection fraction (LVEF) from gated 99mTc-sestamibi myocardial perfusion SPECT images. The algorithm operates in the three-dimensional space and uses gated short-axis image volumes. It segments the left ventricle (LV), estimates and displays endocardial and epicardial surfaces for all gating intervals in the cardiac cycle, calculates the relative left ventricular cavity volumes and derives the global EF from the end-diastolic and end-systolic volume, all without operator interaction. The algorithm for measuring LVEF was tested in 65 clinical patients undergoing 16-interval and 8-interval rest-gated SPECT and validated against first-pass radionuclide ventriculography. Automatic segmentation and contouring of the LV was successful in 65/65 (100%) of the studies. Agreement between EFs measured from 8-interval gated SPECT and EFs calculated from first-pass data was high (y = 2.44 + 1.03x, r = 0.909, p < 0.001, s.e.e. = 6.87). Agreement between EF values measured from 16-interval and 8-interval gated SPECT was excellent (y = -2.7 + 0.97x, r = 0.988, p < 0.001, s.e.e. = 2.65), the latter being on average lower by 3.71 percentage points. Our automatic method is rapid and highly agrees with conventional radionuclide measurements of EF, thus providing clinically useful additional information to complement myocardial perfusion studies.


Clock genes and their genomic distributions in three species of salmonid fishes: Associations with genes regulating sexual maturation and cell cycling.
Clock family genes encode transcription factors that regulate clock-controlled genes and thus regulate many physiological mechanisms/processes in a circadian fashion. Clock1 duplicates and copies of Clock3 and NPAS2-like genes were partially characterized (genomic sequencing) and mapped using family-based indels/SNPs in rainbow trout (RT)(Oncorhynchus mykiss), Arctic charr (AC)(Salvelinus alpinus), and Atlantic salmon (AS)(Salmo salar) mapping panels. Clock1 duplicates mapped to linkage groups RT-8/-24, AC-16/-13 and AS-2/-18. Clock3/NPAS2-like genes mapped to RT-9/-20, AC-20/-43, and AS-5. Most of these linkage group regions containing the Clock gene duplicates were derived from the most recent 4R whole genome duplication event specific to the salmonids. These linkage groups contain quantitative trait loci (QTL) for life history and growth traits (i.e., reproduction and cell cycling). Comparative synteny analyses with other model teleost species reveal a high degree of conservation for genes in these chromosomal regions suggesting that functionally related or co-regulated genes are clustered in syntenic blocks. For example, anti-müllerian hormone (amh), regulating sexual maturation, and ornithine decarboxylase antizymes (oaz1 and oaz2), regulating cell cycling, are contained within these syntenic blocks. Synteny analyses indicate that regions homologous to major life-history QTL regions in salmonids contain many candidate genes that are likely to influence reproduction and cell cycling. The order of these genes is highly conserved across the vertebrate species examined, and as such, these genes may make up a functional cluster of genes that are likely co-regulated. CLOCK, as a transcription factor, is found within this block and therefore has the potential to cis-regulate the processes influenced by these genes. Additionally, clock-controlled genes (CCGs) are located in other life-history QTL regions within salmonids suggesting that at least in part, trans-regulation of these QTL regions may also occur via Clock expression.


Temperamental risk factors for adolescent cannabis use: a systematic review of prospective general population studies.
In order to establish the evidence for prospective relationships between temperamental and personality indicators of behavioral undercontrol and adolescent cannabis use, we systematically searched relevant papers published through April 2008. We assessed and evaluated 14 studies, of which only 4 were considered of high quality. Using "best evidence" synthesis, we found weak to moderate evidence for prospective relations between a combination of high approach and low avoidance and several measures of cannabis use. The study's limitations are noted. This review provides suggestions and recommendations for future studies in this area.


Adverse events of pulmonary vascular stapling in thoracic surgery.
The use of staplers for thoracic surgery has been widely accepted and regarded as a safe procedure. However, adverse events (AEs) of stapling are occasionally experienced. The aim of this retrospective study was to analyse the AEs of stapling in pulmonary vascular surgery. A retrospective multi-institutional review was conducted by the 29 institutions of the Central Japan Lung Cancer Surgery Study Group. All staplings of the pulmonary artery (PA) and vein in thoracic surgery were reviewed during the research period. Stapling of the PA and vein was performed 3393 times. The total number of AEs related to stapling was nine (0.27%). Eight events occurred intraoperatively and one occurred immediately after the operation. Intraoperative AE occurred more frequently than postoperative AE. AE in the PA occurred more frequently than in the pulmonary vein. The intraoperative AEs were oozing (n=3), stapling failure (n=2), laceration of the peripheral vasculature at compression (n=2) and technical injury of the vasculature at insertion (n=1). The causes of AEs were reported to be tissue fragility (n=3), stapler rocking during stapling (n=2), stapler-tissue thickness mismatch (n=2) and technical failure (n=1). The only postoperative AE was staple line rupture of the PV stump. No relationship was seen between the incidence of AE and cartridge colours, compression types of staplers or numbers of staple lines. Generally, stapling of the pulmonary vasculatures in recent thoracic surgery has been safe. Furthermore, the knowledge of the possible risks of pulmonary vascular stapling may help to decrease the AEs of stapling.


Recent applications of electrophysiologic monitoring during surgery.
A multiplicity of electrophysiologic monitoring methods are available to assist the surgeon in operating on a patient in whom there is a risk of damage to neural structures. The applications and problems of each method must be considered in selecting the monitoring technique for each individual patient.


Predation resistance and nematocyst scaling for Metridium senile and M. farcimen.
Previous studies suggest that large body size reduces the risk of predation for acontiate sea anemones. For two species of Metridium, we found significant increases in the length of the acontial threads and in the mean lengths of the unfired acontial nematocyst capsules, with increasing body size. This supports the hypothesis that more damaging acontial defenses protect larger acontiate anemones from their predators. Metridium is planktivorous, and food size does not increase substantially with body size; so we expected smaller increases in nematocyst size for the feeding tentacles. In fact, scaling exponents were significantly smaller for the tentacle nematocysts than for acontial nematocysts of the same types in 3 out of 4 cases. This suggests that nematocyst scaling responds predictably to selection pressure. When specimens of the same size were compared, the non-clonal, subtidal species, M. farcimen, had significantly larger acontial nematocysts than did its clonal congener, M. senile, which lives at the upper tidal limits for major subtidal predators in the northeastern Pacific. Therefore, larger acontial nematocysts may be particularly advantageous where predation levels are high. These data demonstrate that closely related anemone species can be distinguished on the basis of ecologically and functionally relevant differences in nematocyst scaling.


An interaction between systolic blood pressure and angiotensin-converting enzyme gene polymorphism on carotid atherosclerosis.
The insertion/deletion (I/D) polymorphism of the human angiotensin-converting enzyme (ACE) gene is a major determinant of circulating ACE activity, with the D allele being associated with higher ACE levels than the I allele. Thus, chronic exposure to high levels of circulating and tissue ACE may well predispose to vascular wall thickening and atherosclerosis. However, the effect of the ACE gene on carotid atherosclerosis remains controversial. We investigated the association between ACE gene I/D polymorphism and risk factor-dependent augmentation of carotid arterial remodeling in subjects with several risk factors for atherosclerosis. We evaluated sclerotic lesions of the common carotid artery with intima-media thickness (IMT) by ultrasonography in 184 patients (mean age +/- SD, 67 +/- 14 years old) and studied whether any risk factor-gene interactions were associated with carotid atherosclerosis. Out of the 184 subjects, 71 had the ACE II genotype, 87 the ID genotype and 26 the DD genotype. There was no significant difference in IMT among the three ACE genotypes. In total subjects, multiple regression analysis showed that age, total-cholesterol (T-C), and HDL-cholesterol (HDL-C) were significantly associated with IMT. However, the association between risk factors and IMT was genotype-specific. Systolic blood pressure (SBP) and HDL-C were significantly associated with IMT in ACE D carriers (DD+ID), but not in subjects with the ACE II genotype. Similarly, T-C was significantly associated with IMT only in subjects with the ACE II genotype. A general linear model of the interaction between the ACE genotype and the conventional risk factors showed that the SBP-ACE genotype interaction were significantly associated with IMT (F = 7.915; p = 0.005). This finding further supports the idea that analysis of risk factor-gene interaction could be a useful tool for deriving specific predictive information about the development of atherosclerosis.


Composition of myofiber types in the vocalis muscles involved in rapid closure of the glottis in Japanese macaques.
Myofiber types of the medial thyroarytenoid (vocalis) muscle, lateral thyroarytenoid muscle, and cricothyroid muscle of the Japanese macaque were examined with enzyme-histochemical methods. For comparison, the semitendinosus muscle of the Japanese macaque and the thyroarytenoid (vocalis) muscle of cattle, sheep, and pig were examined with the same methods. The vocalis muscle of the Japanese macaque was composed exclusively of fast-twitch/oxidative/glycolytic (FOG) myofibers; it differed from the lateral thyroarytenoid, cricothyroid, and semitendinosus muscles of the Japanese macaque and from the vocalis muscles of the other animals, which consisted of slow-twitch/oxidative, FOG-, and fast-twitch/glycolytic myofibers or type IIC myofibers. The histochemical properties of the vocalis muscle of the Japanese macaque show that the vocalis muscle has a capacity to close the glottis rapidly and completely.


Incipient speciation by sexual isolation in Drosophila melanogaster: extensive genetic divergence without reinforcement.
The collection of Drosophila melanogaster from Zimbabwe and nearby regions (the Z-type) yield females who would not mate with the cosmopolitan D. melanogaster males (the M-type). To dissect the genetic basis of this sexual isolation, we constructed 16 whole-chromosome substitution lines between two standard Z- and M-lines. The results were as follows: (1) All substitution lines appear normal in viability and fertility in both sexes, indicating no strong postmating isolation. (2) The genes for the behaviors are mapped to all three major chromosomes with the same ranking and comparable magnitude of effects for both sexes: III > II >> X > or = 0 (III, II and X designate the effects of the three chromosomes). The results suggest less evolution on the X than on autosomes at loci of sexual behavior. (3) The genes for "Z-maleness" are many and somewhat redundant. Whole-chromosome effects for Z-maleness appear nearly additive and show little dominance. (4) In contrast, "Z-femaleness" has less redundancy as partial genotypes never exhibit full phenotypic effects. Epistatic interactions and incomplete dominance can sometimes be detected. (5) The extensive genetic divergence underlying sexual isolation has evolved in the absence of detectable reduction in hybrid fitnesses. Sexual selection has apparently been a driving force of multiple facets of speciation at the nascent stage without reinforcement.


Teratogenic effect of the cholesterol synthesis inhibitor AY 9944 on rat embryos in vitro.
AY 9944 [trans-1,4-bis(2-chlorobenzylaminomethyl) cyclohexane dihydrochloride] is an amphiphilic cationic molecule. This chemical is an established inhibitor of cholesterol synthesis and is teratogenic in rats. The mechanisms of this teratogenicity remain to be clarified. This study used cultured rat whole embryos to ascertain whether AY 9944 had a direct effect on embryos, or whether its action was indirect, via the maternal cholesterol metabolism. Four experimental conditions were investigated: (A) controls; (B) 10 day untreated embryos were cultured in serum of treated rats; (C) 10 day untreated embryos were cultured in serum containing added AY 9944 (0-1,000 micrograms/ml); and (D) 10 day embryos from females treated on day 4 of gestation were cultured in normal serum. In group B there was no growth retardation; some slight nonspecific abnormalities were not significant. In group C, direct addition of AY 9944 to culture medium retarded growth and differentiation in a dose-dependent manner. No malformation was observed, but histological examinations showed numerous areas of cell necrosis, especially in the CNS. In group D, not only was growth retardation observed, but also characteristic malformations of AY 9944 teratogenesis, including pituitary agenesis. These results show that AY 9944 teratogenicity is initiated prior to day 10.


Massive peri-renal haemorrhage from ruptured renal artery aneurysm in the presence of normal renal function.
A 65-year-old man presented acutely with a large right peri-nephric haematoma as seen on a CT scan. Intravenous urography demonstrated normal function. An arteriogram later revealed a 1 cm renal artery tributary aneurysm, which was successfully embolized.


Serum response elements activate and cAMP responsive elements inhibit expression of transcription factor Egr-1 in synovial fibroblasts of rheumatoid arthritis patients.
Analyzing the induction kinetics and promoter elements regulating the expression of the transcription factor Egr-1, we found elevated levels of Egr-1-encoding mRNA in synovial fibroblasts of rheumatoid arthritis (RA) patients when compared to controls. By contrast, synovial lymphocytes and macrophages do not show an elevated Egr-1 transcription. Therefore, the overexpression of Egr-1 may serve as a diagnostic marker to characterize synovial fibroblasts of RA patients. To study the regulatory mechanisms controlling Egr-1 expression we analyzed the function of transcription factor binding sites located in the Egr-1 promoter. Individual transcription factor binding sites within the Egr-1 promoter were specifically mutated and Egr-1 promoter activity was tested using reporter gene constructs. Our experiments demonstrate that serum response elements are the main positive regulators and binding to a cAMP responsive element represents the major negative regulator for Egr-1 expression in synovial fibroblasts. In addition, we functionally defined a new element, which was not yet described in the human Egr-1 promoter and which serves as a second negative regulatory element for Egr-1 expression. Therefore increased serum response factor activity or failure of Egr-1 repressing signals may account for Egr-1 overexpression in RA synovial fibroblasts.


DFT study of ionic peapod structures from single-walled carbon nanotubes and Lindqvist tungstates.
DFT investigations have been carried out on encapsulation of Lindqvist-type W(6)O(19)(2-) anion inside hydrogenated (n,n) armchair single-walled carbon nanotubes (h-CNTs) with n = 8, 9, 10 to understand the confinement effect of the CNTs on the rotation of W(6)O(19)(2-). The energy-decomposition analysis (EDA) of interaction between W(6)O(19)(2-) and CNTs shows that with the increase of confinement effect from n = 8, 9, to 10, the destabilizing ΔE(Pauli) plays a more important role in the relative orientation of W(6)O(19)(2-) inside CNTs. For W(6)O(19)(2-)@(9,9) h-CNT, the most stable orientation appears at the y/z angle 45°/36°. The confinement effect reduces significantly the energy gap of W(6)O(19)(2-)@(n,n) h-CNT (n = 8, 9, 10) compared with free W(6)O(19)(2-). Electron transfer from the W(6)O(19)(2-) to CNT is observed.


Chicken egg-white extracts promote OCT4 and NANOG expression and telomeres growth in 293T cells.
It will have broad applications in cell biology if one of egg cell extracts has the roles to promote cell proliferation and reprogramming. It will provide a new method for easier reprogramming somatic cells and promote cell proliferation. We found chicken egg-white extracts have roles to promote cell proliferation and reprogramming. The different ingredients were then assessed for cell proliferation activity and somatic cell reprogramming. Chicken egg-white extract ingredients that were less than 3 kDa (LT3K) promoted cell proliferation. Those ingredients that were greater than 3 kDa (GT3K) promoted the increased expression of pluripotency factors in somatic cells and promote telomeres growth in 293T cells. Chicken egg-whites can be separated into ingredients of LT3K, which act to promote cell proliferation, and GT3K, which can be used to promote somatic cell reprogramming.


Isolation of dihydroxyacetone phosphate reductase from dunaliella chloroplasts and comparison with isozymes from spinach leaves.
A dihydroxyacetone phosphate (DHAP) reductase has been isolated in 50% yield from Dunaliella tertiolecta by rapid chromatography on diethylaminoethyl cellulose. The activity was located in the chloroplasts. The enzyme was cold labile, but if stored with 2 molar glycerol, most of the activity was restored at 30 degrees C after 20 minutes. The spinach (Spinacia oleracea L.) reductase isoforms were not activated by heat treatment. Whereas the spinach chloroplast DHAP reductase isoform was stimulated by leaf thioredoxin, the enzyme from Dunaliella was stimulated by reduced Escherichia coli thioredoxin. The reductase from Dunaliella was insensitive to surfactants, whereas the higher plant reductases were completely inhibited by traces of detergents. The partially purified, cold-inactivated reductase from Dunaliella was reactivated and stimulated by 25 millimolar Mg(2+) or by 250 millimolar salts, such as NaCl or KCl, which inhibited the spinach chloroplast enzyme. Phosphate at 3 to 10 millimolar severely inhibited the algal enzyme, whereas phosphate stimulated the isoform in spinach chloroplasts. Phosphate inhibition of the algal reductase was partially reversed by the addition of NaCl or MgCl(2) and totally by both. In the presence of 10 millimolar phosphate, 25 millimolar MgCl(2), and 100 millimolar NaCl, reduced thioredoxin causes a further twofold stimulation of the algal enzyme. The Dunaliella reductase utilized either NADH or NADPH with the same pH maximum at about 7.0. The apparent K(m) (NADH) was 74 micromolar and K(m) (NADPH) was 81 micromolar. Apparent V(max) was 1100 mumoles DHAP reduced per hour per milligram chlorophyll for NADH, but due to NADH inhibition highest measured values were 350 to 400. The DHAP reductase from spinach chloroplasts exhibited little activity with NADPH above pH 7.0. Thus, the spinach chloroplast enzyme appears to use NADH in vivo, whereas the chloroplast enzyme from Dunaliella or the cytosolic isozyme from spinach may utilize either nucleotide.


Phosphonoformate: a minimal transition state analogue inhibitor of the fosfomycin resistance protein, FosA.
Fosfomycin [(1R,2S)-epoxypropylphosphonic acid] is a simple phosphonate found to have antibacterial activity against both Gram-positive and Gram-negative microorganisms. Early resistance to the clinical use of the antibiotic was linked to a plasmid-encoded resistance protein, FosA, that catalyzes the addition of glutathione to the oxirane ring, rendering the antibiotic inactive. Subsequent studies led to the discovery of a genomically encoded homologue in the pathogen Pseudomonas aeruginosa. The proteins are Mn(II)-dependent enzymes where the metal is proposed to act as a Lewis acid stabilizing the negative charge that develops on the oxirane oxygen in the transition state. Several simple phosphonates, including the antiviral compound phosphonoformate (K(i) = 0.4 +/- 0.1 microM, K(d) approximately 0.2 microM), are shown to be inhibitors of FosA. The crystal structure of FosA from P. aeruginosa with phosphonoformate bound in the active site has been determined at 0.95 A resolution and reveals that the inhibitor forms a five-coordinate complex with the Mn(II) center with a geometry similar to that proposed for the transition state of the reaction. Binding studies show that phosphonoformate has a near-diffusion-controlled on rate (k(on) approximately 10(7)-10(8) M(-1) s(-1)) and an off rate (k(off) = 5 s(-1)) that is slower than that for fosfomycin (k(off) = 30 s(-1)). Taken together, these data suggest that the FosA-catalyzed reaction has a very early transition state and phosphonoformate acts as a minimal transition state analogue inhibitor.