The impact of the Lisbon Football Derby on the profile of emergency department admissions.
Variations in emergency department admissions have been reported to happen as a result of major sports events. The work presented assessed changes in volume and urgency level of visits to a major Emergency Department in Lisbon during and after the city's football derby. Volume of attendances and patient urgency level, according to the Manchester Triage System, were retrospectively analyzed for the 2008-2011 period. Data regarding 24-hour periods starting 45 minutes before kick-off was collected, along with data from similar periods on the corresponding weekdays in the previous years, to be used as controls. Data samples were organized according to time frame (during and after the match), urgency level, and paired accordingly. A total of 14 relevant periods (7 match and 7 non-match) were analyzed, corresponding to a total of 5861 admissions. During the match time frame, a 20.6% reduction (p = 0.06) in the total number of attendances was found when compared to non-match days. MTS urgency level sub-analysis only showed a statistically significant reduction (26.5%; p = 0.05) in less urgent admissions (triage levels green-blue). Compared to controls, post-match time frames showed a global increase in admissions (5.6%; p = 0.45), significant only when considering less urgent ones (18.9%; p = 0.05). A decrease in the total number of emergency department attendances occurred during the matches, followed by a subsequent increase in the following hours. These variations only reached significance among visits triaged green-blue. During major sports events an overall decrease in emergency department admissions seems to take place, especially due to a drop in visits associated with less severe conditions.


The effects of extra high dose rate irradiation on glioma stem-like cells.
Radiation therapy is an integral part of treatment for patients with glioblastoma. New technological advances in linear accelerators have made extra-high dose rate irradiation possible. This shortens patient treatment time significantly compared to standard dose rate irradiation, but the biologic effects of extra high dose rate irradiation are poorly understood. Glioma stem-like cells (GSCs) are resistant to standard radiation and contribute to tumor progression. Here, we assess the therapeutic effect of extra high dose rate vs. standard dose rate irradiation on GSCs. GSCs were exposed to 2, 4 and 6 Gy X-irradiation at dose rates of 4.2 Gy/min or 21.2 Gy/min (400 monitoring units (MU)/min or 2100 MU/min). We analyzed cell survival with cell growth assays, tumorsphere formation assays and colony formation assays. Cell kill and self-renewal were dependent on the total dose of radiation delivered. However, there was no difference in survival of GSCs or DNA damage repair in GSCs irradiated at different dose rates. GSCs exhibited significant G1 and G2/M phase arrest and increased apoptosis with higher doses of radiation but there was no difference between the two dose rates at each given dose. In a GSC-derived preclinical model of glioblastoma, radiation extended animal survival, but there was no difference in survival in mice receiving different dose rates of radiation. We conclude that GSCs respond to larger fractions of radiation, but extra high dose rate irradiation has no significant biologic advantage in comparison with standard dose rate irradiation.


[Chemical integration in mechanisms of brain plasticity].
The paper analyzes investigations on brain functions from the viewpoint of P. K. Anokhin's theory of functional systems. Brain plasticity is postulated to be the result of convergence of various excitations on single brain neurons and of chemical integration of brain transmitters and modulators of peptide origin in the realization of biological motivations. Numerous findings of duplication of various neurotransmitters and oligopeptides in the central mechanisms of biological motivations, chemical rearrangement in the recovery period after the lateral hypothalamic syndrome when bilateral motivational centers of the hypothalamus were coagulated suggests that there is chemical compensation in the brain activity for homeostatic maintenance. The plasticity of extrahypothalamic brain structures are postulated to be more expressed as compared to the specific activities of hypothalamic, brain stem, and spinal cord structures.


The good, the bad, and the cell type-specific roles of hypoxia inducible factor-1 alpha in neurons and astrocytes.
Hypoxia inducible factor-1alpha (HIF-1alpha) is a key regulator of oxygen homeostasis, because it is responsible for the regulation of genes involved in glycolysis, erythropoiesis, angiogenesis, and apoptosis. In the CNS, HIF-1alpha is stabilized by insults associated with hypoxia and ischemia. Because its many target genes mediate both adaptive and pathological processes, the role of HIF-1alpha in neuronal survival is debated. Although neuronal HIF-1alpha function has been the topic of several studies, the role of HIF-1alpha function in astrocytes has received much less attention. To characterize the role of HIF-1alpha in neurons and astrocytes, we induced loss of HIF-1alpha function specifically in neurons, astrocytes, or both cell types in neuron/astrocyte cocultures exposed to hypoxia. Although loss of HIF-1alpha function in neurons reduced neuronal viability during hypoxia, selective loss of HIF-1 function in astrocytes markedly protected neurons from hypoxic-induced neuronal death. Although the pathological processes induced by HIF-1alpha in astrocytes remain to be defined, induction of inducible nitric oxide synthase likely contributes to the pathological process. This study delineates, for the first time, a cell type-specific action for HIF-1alpha within astrocytes and neurons.


Randomized, double-blind, placebo-controlled trial of the 5-HT1A receptor antagonist AZD7371 tartrate monohydrate (robalzotan tartrate monohydrate) in patients with irritable bowel syndrome.
To investigate the efficacy and safety of the 5-hydroxytrypamine 1A (5-HT(1A)) receptor antagonist AZD7371 tartrate monohydrate (robalzotan tartrate monohydrate), termed AZD7371 here, in patients with irritable bowel syndrome (IBS). Patients meeting the Rome II criteria for IBS (N = 402) were randomized to treatment with AZD7371 20 mg or 5 mg or matching placebo tablets twice daily for 12 wk. The patients completed daily and weekly diary assessments, reporting abdominal discomfort or pain and description of bowel movements. They also completed validated symptom and quality-of-life questionnaires. Neither AZD7371 regimen was significantly more effective than placebo in providing adequate relief from IBS symptoms in at least 2 out of 4 wk per month over the 12 wk of treatment. There was also no significant difference between the treatment groups and placebo in the change in score in the validated symptom and quality-of-life questionnaires. Overall, 22.1% of patients experienced adverse events (AEs) attributed to the study medication: 44 of 133 (33.1%) in the 20 mg AZD7371 group, 27 of 131 (20.6%) in the 5 mg AZD7371 group, and 17 of 134 (12.7%) in the placebo group. Also, 31 of 57 (54%) of AEs leading to discontinuation were central nervous system-related. Hallucinations or hallucination-like AEs were reported by eight patients taking AZD7371, and by none of the patients in the placebo group. After these events led to discontinuation in six patients, the study was prematurely terminated. In view of the AE profile and lack of efficacy in IBS, the clinical development of AZD7371 has been stopped.


Quinine, an old anti-malarial drug in a modern world: role in the treatment of malaria.
Quinine remains an important anti-malarial drug almost 400 years after its effectiveness was first documented. However, its continued use is challenged by its poor tolerability, poor compliance with complex dosing regimens, and the availability of more efficacious anti-malarial drugs. This article reviews the historical role of quinine, considers its current usage and provides insight into its appropriate future use in the treatment of malaria. In light of recent research findings intravenous artesunate should be the first-line drug for severe malaria, with quinine as an alternative. The role of rectal quinine as pre-referral treatment for severe malaria has not been fully explored, but it remains a promising intervention. In pregnancy, quinine continues to play a critical role in the management of malaria, especially in the first trimester, and it will remain a mainstay of treatment until safer alternatives become available. For uncomplicated malaria, artemisinin-based combination therapy (ACT) offers a better option than quinine though the difficulty of maintaining a steady supply of ACT in resource-limited settings renders the rapid withdrawal of quinine for uncomplicated malaria cases risky. The best approach would be to identify solutions to ACT stock-outs, maintain quinine in case of ACT stock-outs, and evaluate strategies for improving quinine treatment outcomes by combining it with antibiotics. In HIV and TB infected populations, concerns about potential interactions between quinine and antiretroviral and anti-tuberculosis drugs exist, and these will need further research and pharmacovigilance.


Early discoveries on enzyme deficiencies in lysosomal storage diseases: The Indian contribution.
The discovery of enzyme deficiencies in lysosomal storage disorders began with two discoveries made in 1963. One of these was made by a Belgian scientist, Henri-Gery Hers, who discovered that in Pompe's disease there was a deficiency in α-glucosidase. The other was made by an international collaboration involving an American neurologist, James Austin, and an Indian biochemist, Bimal Bachhawat, where the enzyme arylsulfatase A was found deficient in metachromatic leukodystrophy. This article attempts to trace the events that led to this fruitful collaboration and how these two young investigators eventually discovered the defective enzyme in metachromatic leukodystrophy.


Root resorption in elderly patients.
Root resorption in permanent teeth is a frequently observed pathology that may originate in various causes. Life expectancy is progressively rising, odontological preventive care is becoming more widespread and professionals are educating their patients in the importance of preventive practices. Because senior citizens are thus losing fewer teeth prematurely they will be conversely more at risk for dental problems later in life. The knowledge of the alterations that may appear in the roots of geriatric patients is particularly relevant to devising therapy and establishing prognosis. The aim of the present study was to evaluate the nature and magnitude of the histologic and histomorphometric features of root resorption and the eventual possibility of repair in elderly people. Seventy-seven uniradicular teeth of patients aged between 65 and 90 years and 18 premolars of patients aged between 14 and 20 years, were removed, fixed in 90% formalin, decalcified in EDTA and embedded in paraffin. Vestibulo-lingual sections were stained with hematoxylin-eosin and employed to perform histological and histomorphometric studies. The results showed that 30% of the teeth of younger patients and 94% of the teeth of elderly patients exhibited areas of root resorption. From the 416 resorptive areas found in elderly patients, 173 exhibited signs of repair being the volume/surface ratio of these areas 0.69 +/- 0.06. These data show that root resorption is a frequent finding in the older population under study. Resorptions are characterized by scarce depth, large areas and a high incidence of repair despite the old age of the patients.


Rosmarinic acid counteracts activation of hepatic stellate cells via inhibiting the ROS-dependent MMP-2 activity: Involvement of Nrf2 antioxidant system.
Recently, oxidative stress is involved in hepatofibrogenesis. Matrix metalloproteinase-2 (MMP-2) is required for activation of hepatic stellate cells (HSCs) in response to reactive oxygen species (ROS). This study was designed to explore the hypothesis that the inhibitory effect of rosmarinic acid (RA) on HSCs activation might mainly result from its antioxidant capability by increasing the synthesis of glutathione (GSH) involved in nuclear factor kappa B (NF-κB)-dependent inhibition of MMP-2 activity. Here, we demonstrate that RA reverses activated HSCs to quiescent cells. Concomitantly, RA inhibits MMP-2 activity. RNA interference-imposed knockdown of NF-κB abolished down-regulation of MMP-2 by RA. RA-mediated inactivation of NF-κB could be blocked by the diphenyleneiodonium chloride (DPI; a ROS inhibitor). Conversely, transfection of dominant-negative (DN) mutant of extracellular signal-regulated kinases 2 (ERK2), c-Jun N-terminal kinase 1 (JNK1), or p38α kinase had no such effect. Simultaneously, RA suppresses ROS generation and lipid peroxidation (LPO) whereas increases cellular GSH in HSC-T6 cells. Furthermore, RA significantly increased antioxidant response element (ARE)-mediated luciferase activity, nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and catalytic subunits from glutamate cysteine ligase (GCLc) expression, but not modulatory subunits from GCL (GCLm). RA-mediated up-regulation of GClc is inhibited by the shRNA-induced Nrf2 knockdown. The knocking down of Nrf2 or buthionine sulfoximine (a GCL inhibitor) abolished RA-mediated inhibition of ROS. Collectively, these results provide novel insights into the mechanisms of RA as an antifibrogenic candidate in the prevention and treatment of liver fibrosis.


An optimized HPLC-UV method for quantitatively determining sesquiterpenes in Nardostachyos Radix et Rhizoma.
Nardostachyos Radix et Rhizoma (NR), the root and rhizome from either Nardostachys jatamansi Batal or Nardostachys jatamansi DC, is known to have biological functions including neuro-protective and anti-pancreatitis activity. The main bioactive compounds within NR are all classified as sesquiterpenes, and include desoxo-narchinol A, nardosinonediol, and nardosinone. Although NR is a valuable herb that is widely used in many Asian countries, robust quality control protocols for NR are still in question, especially those that can analyze the three main active compounds. Current quantitative methods within the Chinese Pharmacopoeic use nardosinone as a marker compounds. One compound cannot represent a complicated matrix, and is thus insufficient to control the quality of this herbal medicine. Moreover, there are no high-performance liquid chromatography (HPLC) methods that can simultaneously analyze desoxo-narchinol A (DA), nardosinonediol (NE), and nardosinone (ND) within NR. This study aimed to establish an efficient quality control protocol by developing an analytical method that simultaneously detects the three sesquiterpenes with HPLC using response surface methodology (RSM) to optimize sample preparation. Optimized HPLC conditions included a mobile phase of 0.1% formic acid in water (A), and a 0.1% formic acid in acetonitrile (B) under an elution program of 20% B-80% B for 30min at 254nm. The method was well validated, demonstrating satisfactory linearity, accuracy, precision, recovery, repeatability, and stability. Optimized conditions for creating the analytical sample were predicted by RSM using a Box-Behnken design. These conditions included reflux at 70°C for 3h using 24.98% ethanol as the extraction solvent (solvent: solid ratio=78.81mL/g). The relationship between the results between predicted and experimental conditions was well correlated, and varied between 96.48%-102.11%. Thus, our developed HPLC method, paired with optimized sample preparation conditions, accurately quantified all three sesquiterpenes, and may thus be a prospective means of controlling the quality of NR.


Comparative Monte-Carlo simulations of charge carrier transport in amorphous molecular solids as given by three most common models of disorder: The dipolar glass, the Gaussian disorder, and their mix.
We have performed Monte-Carlo simulations of the charge carrier transport in a model molecularly doped polymer using three most popular hopping theories (the dipolar glass model, the Gaussian disorder model, and an intermediate between them) in a wide range of applied electric fields and temperatures. Time of flight transients have been computed and analyzed in logarithmic coordinates to study the Poole-Frenkel field dependence, the non-Arrhenius mobility temperature dependence, and the nondispersive versus dispersive current shapes. We also have made an attempt to estimate the total disorder energy directly from simulation data at the lowest electric field thus checking the consistency of the model fitting. Computational results have been compared with the analytical and experimental information available in the literature.


The use of tailored subheadings was successful in enhancing compliance with CONSORT in a dental journal.
Efforts to enhance the reporting of clinical trials have intensified in recent years with automated strategies and editorial involvement showing promise in improving compliance with accepted guidelines. This study aimed to evaluate the effectiveness of a concerted approach to adherence to CONSORT (CONsolidated Standards Of Reporting Trials) guidelines in a dental journal. Following the publication of an exemplar clinical trial on the American Journal of Orthodontics and Dentofacial Orthopedics (AJO-DO) website and related changes to the author guidelines, trial submissions were required to follow a standard format incorporating subheadings mirroring the CONSORT guidelines. Compliance with CONSORT was assessed in initial submissions over a 30-month period. Reporting was compared to submissions of randomized controlled trials (RCTs) which did not include subheadings over the same period. Seventy-one RCTs were submitted to the AJO-DO from January 2014 to June 2016, 49 with subheadings and 22 without. Most CONSORT items (e.g. random sequence generation, allocation concealment and blinding) were more frequently adequately reported when RCTs were submitted with inclusion of subheadings. Overall, reporting quality of the submitted RCTs was 15.2% higher with use of the subheadings format (95%CI: 10.5, 20.0; p<0.001) with a mean overall score of 87.3%. Enhanced compliance of submitted RCTs was found with use of a bespoke approach to trial presentation utilizing CONSORT item subheadings. The improvement in initial submissions is particularly encouraging as this arose without input either from peer reviewers or journal editors. This simple approach may have wider applicability.


Interspecific cell markers and cell lineage in birds.
A cell marking technique based on the structural differences existing between the interphase nucleus in two closely related species of birds, the chick and the Japanese quail, is described. In all embryonic and adult cell types of the quail, a large mass of heterochromatin is associated with the nucleolus making quail and chick cells easy to identify at the single cell level after application of any DNA-specific staining procedure and also at the electron microscope level. This method has been largely used to construct chimeras in ovo and to study dynamic processes such as cell migrations or cell lineage segregation during ontogeny. Recently monoclonal antibodies specific for either quail or chick antigenic determinants (for example, class II MHC antigens) have been prepared, increasing the interest of the quail-chick chimera system as an experimental model.


The change of prevalence and recurrence of acute otitis media in Korea.
Acute otitis media (AOM) is the most common infectious disease in infants and children. However, there have been few studies on the recent prevalence and trends of pediatric AOM in Korea. This study aimed to evaluate the prevalence and trends of pediatric AOM in Korea. The data for patients with AOM in 2012, 2015, and 2017 were extracted from the Health Insurance Review and Assessment Service's Big Database. The prevalence rate, recurrence rate, and data on gender, region, month, and medical facilities of AOM were evaluated. The prevalence rate (per 1000 persons) decreased from 152.7 in 2012 to 137.4 in 2017. The recurrence rate declined from 34.0% in 2012 to 28.2% in 2015. In 2015, the recurrence rate of 0-2 years group decreased significantly. In 2017, the prevalence of AOM of 3-6 years group decreased the most. The prevalence is higher in urban areas and recurrence rate is higher in rural areas. But those are no significant difference. AOM is more common in male, mainly in winter and spring. The prevalence and recurrence rate of pediatric AOM in Korea are gradually decreasing. The decrease of prevalence resulted from declination in AOM recurrence of 0-2 years group in 2015 and AOM occurrence of 3-6 years group in 2017.


A BRCA1 Coiled-Coil Domain Variant Disrupting PALB2 Interaction Promotes the Development of Mammary Tumors and Confers a Targetable Defect in Homologous Recombination Repair.
The BRCA1 tumor suppressor gene encodes a multidomain protein for which several functions have been described. These include a key role in homologous recombination repair (HRR) of DNA double-strand breaks, which is shared with two other high-risk hereditary breast cancer suppressors, BRCA2 and PALB2. Although both BRCA1 and BRCA2 interact with PALB2, BRCA1 missense variants affecting its PALB2-interacting coiled-coil domain are considered variants of uncertain clinical significance (VUS). Using genetically engineered mice, we show here that a BRCA1 coiled-coil domain VUS, Brca1 p.L1363P, disrupts the interaction with PALB2 and leads to embryonic lethality. Brca1 p.L1363P led to a similar acceleration in the development of Trp53-deficient mammary tumors as Brca1 loss, but the tumors showed distinct histopathologic features, with more stable DNA copy number profiles in Brca1 p.L1363P tumors. Nevertheless, Brca1 p.L1363P mammary tumors were HRR incompetent and responsive to cisplatin and PARP inhibition. Overall, these results provide the first direct evidence that a BRCA1 missense variant outside of the RING and BRCT domains increases the risk of breast cancer. SIGNIFICANCE: These findings reveal the importance of a patient-derived BRCA1 coiled-coil domain sequence variant in embryonic development, mammary tumor suppression, and therapy response.See related commentary by Mishra et al., p. 6080.


Microbial Pathogens at the Movies.
Usually, show business depicts viruses, bacteria, and other microorganisms as one of the worse menaces to mankind. Entertainment movies influence the way audiences understand and perceive these topics. Few films accurately portray the science of microbiology and its social implications. Movies and TV series often feature outbreaks of deadly diseases and the efforts of scientists and medical professionals to contain them. But entertainment movies can also be used to educate the public about the importance and the impact that microorganisms have on our lives, helping to increase public awareness and appreciation of the world of microbiology. The aim of this review is to show the relationship between movies and microbiology, from the fight against diseases such as AIDS or tuberculosis, to the zombie apocalypse.


IgE receptors and serum IgE in pollinosis. The influence of season and action of ketotifen.
In the present study we have analyzed the correlation between bound and free IgE in two groups of patients with sensitization to grass pollen; 15 of these patients only presented symptoms of rhinitis, while the other 17 patients had symptoms associated with bronchial asthma. The study was carried out after, and during the pollen season. We observed that no correlation existed between bound and free IgE in the rhinitic group after pollen season, while there was correlation with the same group of patients during the pollen season. On the contrary, the asthmatic group obtained a good correlation between bound and free IgE after the pollen season, rather than within the pollinization period. In view of these results, we can affirm that the relationship which exists between free and membrane-associated IgE varies, depending on the type of individual and the seasonal period at the time the measurements were carried out. At the same time and in order to analyze the dynamics of action of ketotifen, we studied the effect of this drug on the amount of membrane-associated IgE, for which we employed an enriched population of basophils maintained in culture. From the results obtained we deduced that in no way does ketotifen affect the receptor expression for IgE. Therefore, the possible mechanism of action of ketotifen could be directed toward the interior of, rather than the exterior of the plasmatic membrane.


New coagulant agent (ankaferd blood stopper) for open hemorrhages in hemophilia with inhibitor.
The treatment of hemophilia A patients with inhibitor could be very expensive. Ankaferd blood stopper (ABS) is a unique folkloric medicinal plant extract, which has historically been used in Turkish traditional medicine as a hemostatic agent. In this article, a 16-year-old boy was presented with uncontrolled bleeding, despite the treatment of factor VIII, rVIIa, factor VIII inhibitor bypass activity (FEIBA), cyclophosphamide, and prednisolone at circumcision site that resolved with ABS in minutes. Our patient with hemophilia A and inhibitor is the first clinical pediatric case.


Distribution and Location of BEVs in Different Genotypes of Bananas Reveal the Coevolution of BSVs and Bananas.
Members of the family Caulimoviridae contain abundant endogenous pararetroviral sequences (EPRVs) integrated into the host genome. Banana streak virus (BSV), a member of the genus Badnavirus in this family, has two distinct badnaviral integrated sequences, endogenous BSV (eBSV) and banana endogenous badnavirus sequences (BEVs). BEVs are distributed widely across the genomes of different genotypes of bananas. To clarify the distribution and location of BEVs in different genotypes of bananas and their coevolutionary relationship with bananas and BSVs, BEVs and BSVs were identified in 102 collected banana samples, and a total of 327 BEVs were obtained and categorized into 26 BEVs species with different detection rates. However, the majority of BEVs were found in Clade II, and a few were clustered in Clade I. Additionally, BEVs and BSVs shared five common conserved motifs. However, BEVs had two unique amino acids, methionine and lysine, which differed from BSVs. BEVs were distributed unequally on most of chromosomes and formed hotspots. Interestingly, a colinear relationship of BEVs was found between AA and BB, as well as AA and SS genotypes of bananas. Notably, the chromosome integration time of different BEVs varied. Based on our findings, we propose that the coevolution of bananas and BSVs is driven by BSV Driving Force (BDF), a complex interaction between BSVs, eBSVs, and BEVs. This study provides the first clarification of the relationship between BEVs and the coevolution of BSVs and bananas in China.


[Spontaneous hepatitis A with a fatal outcome in rhesus monkeys].
This is the first report of virologically verified spontaneous hepatitis A in M. rhesus monkeys with severe involvement of the liver leading to the death of the animals. In 21 out of 23 dead monkeys morphological lesions in the liver have been characterized as acute hepatitis. In 6 (26%) animals no other pathological processes were found. In 15 animals hepatitis was combined with other diseases (dysentery, parasitic infestations, coronavirus infection). Antigen of hepatitis A virus was detected by an enzyme immunoassay in the intestinal contents of 8 monkeys and in the livers of 3 of them. Immune electron microscopic studies detected in the intestines some virus particles morphologically and antigenically similar to human hepatitis A virus.


Color Doppler-An effective tool for diagnosing midgut volvulus with malrotation.
Malrotation with midgut volvulus is a common surgical emergency in children. A diagnostic tool for malrotation with characteristics ideal in emergency settings such as non-invasiveness and rapidity remains a keenly debated issue among surgeons and radiologists alike. Fifty-two pediatric patients of suspected malrotation with midgut volvulus were studied between 1998 and 2016. All patients underwent plain abdominal X-ray and Color Doppler; contrast upper gastrointestinal (GI) studywas done in some patients. All cases were operated and diagnosis was confirmed. A subset of 60 pediatric patients with nonspecific GI complaints were also examined to see relative position of superior mesenteric vein (SMV)/superior mesenteric artery (SMA) in control population. A total of 52 suspected cases of malrotation were admitted from May 1998 to November 2015, 43 had inversion of SMA/SMV and nine had SMV anterior of SMA in Color Doppler. All 43 cases of inversion of SMA/SMV were cases of malrotation after surgical confirmation; while five out of nine cases of SMV anterior to SMA had malrotation. In appropriate clinical settings, Color Doppler documenting the reversal or aberrant SMV/SMA axis is not only predictive but also diagnostic of malrotation of gut.


LacZ expression in Fut2-LacZ reporter mice reveals estrogen-regulated endocervical glandular expression during estrous cycle, hormone replacement, and pregnancy.
The secretor gene (FUT2) encodes an alpha(1,2)fucosyltransferase (E.C. 18.104.22.168) that elaborates alpha(1,2)fucose residues on mucosal epithelium and secreted mucins. Though uterine alpha(1,2)fucosylated glycans have been proposed to be involved in embryo adhesion, mice with a homozygous null mutation of Fut2 displayed normal fertility. To help develop alternative hypotheses for function, the cell type and regulation of Fut2 expression during the estrous cycle, hormone replacement, and pregnancy was examined in Fut2-LacZ reporter mice containing targeted replacement of Fut2 with bacterial lacZ. LacZ expression in the reproductive tract of Fut2-LacZ mice is most prominent in the glandular epithelium of the endocervix during estrus and pregnancy. Nuclear LacZ expression identifies cell-specific expression of Fut2 in mucus-secreting cells of the endocervix, uterine glands, foveolar pit and chief cells of the stomach, and goblet cells of the colon. In ovariectomized Fut2-LacZ mice, estradiol treatment stimulates X-gal staining in endocervix and uterus but does not affect expression in stomach and colon. Northern blot analysis in wild-type mice shows 15-fold elevations of Fut2 steady-state mRNA with estradiol treatment, whereas Fut1 varies little. Fut2 levels in the glandular stomach and distal colon remain constant, and uterine Fut2 levels vary eightfold during the estrous cycle. These data represent the first demonstration of a glycosyltransferase gene under tissue-specific hormonal regulation in a LacZ reporter mouse model. Endocervical expression of Fut2 in estrus and pregnancy may modify cervical mucus barrier properties from microbial infection analogous to the potential role of mucosal glycans in humans.


A proteomic approach in investigating the hepatoprotective mechanism of Schisandrin B: role of raf kinase inhibitor protein.
To identify key proteins involved in the hepatoprotection afforded by schisandrin B (Sch B), we used a proteomic approach to screen proteins that were specifically regulated by Sch B in mouse livers and to investigate the role of the proteins in hepatoprotection. Thirteen proteins were specifically activated or suppressed by Sch B treatment. Among the 13 proteins, Raf kinase inhibitor protein (RKIP) was postulated to be the key regulator involved in the development of hepatotoxin-induced cellular damage. The results indicated that the downregulation of RKIP by antisense RKIP vector transfection led to the activation of the Raf-1/MEK/ERK signaling pathway, as evidenced by increases in the level of MEK/ERK phosphorylation and the level of nuclear factor erythroid 2-related factor 2 in the nucleus. The signaling effect produced by RKIP downregulation resembled that triggered by Sch B, wherein both treatments resulted in a decrease in the extent of carbon tetrachloride-induced apoptotic cell death in AML12 hepatocytes. Overexpression of RKIP by the sense RKIP transfection vector or the inhibition of MEK kinase by PD98059 was able to abrogate the cytoprotective effect of Sch B in the hepatocytes. The results indicate that Sch B triggers the Raf/MEK/ERK signaling pathway, presumably by downregulating RKIP, thereby protecting against carbon tetrachloride-induced cytotoxicity.


Gene cloning and characterization of alpha-glucuronidase of Bacillus stearothermophilus no. 236.
The alpha-glucuronidase gene of Bacillus stearothermophilus No. 236 was cloned, sequenced, and expressed in Escherichia coli. The gene, designated aguA, encoded a 691-residue polypeptide with calculated molecular weight of 78,156 and pI of 5.34. The alpha-glucuronidase produced by a recombinant E. coli strain containing the aguA gene was purified to apparent homogeneity and characterized. The molecular weight of the alpha-glucuronidase was 77,000 by SDS-PAGE and 161,000 by gel filtration; the functional form of the alpha-glucuronidase therefore was dimeric. The optimal pH and temperature for the enzyme activity were pH 6.5 and 40 degrees C, respectively. The enzyme's half-life at 50 degrees C was 50 min. The values for the kinetic parameters of Km and Vmax were 0.78 mM and 15.3 U/mg for aldotriouronic acid [2-O-alpha-(4-O-methyl-alpha-D-glucopyranosyluronic)-D-xylobiose]. The alpha-glucuronidase acted mainly on small substituted xylo-oligomers and did not release methylglucuronic acid from intact xylan. Nevertheless, synergism in the release of xylose from xylan was found when alpha-glucuronidase was added to a mixture of endoxylanase and beta-xylosidase.


Three-dimensional kinematic analysis of the pectoral girdle during upside-down locomotion of two-toed sloths (Choloepus didactylus, Linné 1758).
Theria (marsupials and placental mammals) are characterized by a highly mobile pectoral girdle in which the scapula has been shown to be an important propulsive element during locomotion. Shoulder function and kinematics are highly conservative during locomotion within quadrupedal therian mammals. In order to gain insight into the functional morphology and evolution of the pectoral girdle of the two-toed sloth we here analyze the anatomy and the three-dimensional (3D) pattern of shoulder kinematics during quadrupedal suspensory ('upside-down') locomotion. We use scientific rotoscoping, a new, non-invasive, markerless approach for x-ray reconstruction of moving morphology (XROMM), to quantify in vivo the 3D movements of all constituent skeletal elements of the shoulder girdle. Additionally we use histologic staining to analyze the configuration of the sterno-clavicular articulation (SCA). Despite the inverse orientation of the body towards gravity, sloths display a 3D kinematic pattern and an orientation of the scapula relative to the thorax similar to pronograde claviculate mammalian species that differs from that of aclaviculate as well as brachiating mammals. Reduction of the relative length of the scapula alters its displacing effect on limb excursions. The configuration of the SCA maximizes mobility at this joint and demonstrates a tensile loading regime between thorax and limbs. The morphological characteristics of the scapula and the SCA allow maximal mobility of the forelimb to facilitate effective locomotion within a discontinuous habitat. These evolutionary changes associated with the adoption of the suspensory posture emphasized humeral influence on forelimb motion, but allowed the retention of the plesiomorphic 3D kinematic pattern.


Helicobacter pylori in the dental plaque and gastric mucosa of dyspeptic Nigerian patients.
Presence of helicobacter pylori (H.Pylori) in the dental plaques may be associated with the presence of the same organism in gastric mucosa. To assess and compare the prevalence of H. pylori in dental plaques and gastric mucosa of dyspeptic Nigerians. Sixty-six consecutive patients undergoing endoscopy for investigation of dyspepsia were included in the study. A predesigned questionnaire was used to collect information on socio-demographic and other factors. Each patient also had clinical dental examination. Dental plaque scrapping was collected before endoscopy for helicobacter pylori culture, and at endoscopy, two gastric biopsy specimens were taken for culture and histology. The mean age (+/- SD) of the patients was 44.8 (+/- 17.4) years (range 5 to 80 years). The positivity rate of helicobacter pylori in gastric biopsy was 35 of 66 (53.0%) while the same for dental plaque was 46 of 66 (69.7%). The correlation (Spearman's) between gastric mucosa and dental plaque colonization with helicobactor pylori was significant (R = 0.30, P = 0.01). When concordant positivity of gastric biopsy culture and histology was taken as diagnostic, the sensitivity of dental plaque culture was found to be 82.9% and the specificity was 45.2%. This study revealed that helicobactor pylori has a high prevalence in the dental plaque of the population that we have studied and may therefore be of potential relevance in screening for such infection.


Faceting on He crystals.
It has been predicted by Landau that, ideally at low temperatures, crystals should show many different types of facets, i.e., flat smooth faces on their surface, but this so-called "devil's staircase" phenomenon has been difficult to observe experimentally. In this paper we describe our recent experiments, in which altogether 11 different types of facets have been identified on growing (3)He crystals at the temperature of 0.55 mK by using a unique low-temperature Fabry-Pérot interferometer. Previously only 3 types of facets had been seen in this system. We have also measured the growth velocities of different facets, and our interpretation of the obtained results yields the conclusion that (3)He has much stronger coupling of the liquid-solid interface to the crystal lattice than has been expected. After an introduction we present a short theoretical background about the equilibrium crystal shape and the roughening transitions, which is followed by the description of our experimental results and discussion.


iCareTrack: measuring the appropriateness of eyecare delivery in Australia.
To meet the needs of an ageing population and optimise health expenditure, delivery of care should be based on evidence. However, the level of evidence-based care delivered to patients with eye conditions is rarely assessed. This study thus aimed to determine the percentage of eyecare encounters at which a sample of adult Australians received appropriate care (i.e., eyecare in line with evidence-based or consensus-based guidelines). A cross-sectional retrospective review of optometry practice records was conducted using random stratified (by state) sampling in mainland Australia. Eighty-five clinical indicators were developed from evidence-based clinical practice guideline recommendations and refined by panels of experts using a modified Delphi process. Healthcare records of patients 18 years and over were examined against these indicators, representing appropriate care for three common eye conditions (preventative eyecare, glaucoma, and diabetic eyecare). Encounters occurred in optometry practices that were selected to be representative of the socioeconomic profile of Australian practices. The primary outcome measure was percentage compliance of eyecare delivery against the clinical indicators. From 426 optometry practices contacted by mail or telephone, 90 (21%) replied, 46 proved eligible and 42 were included in the study and visited for data collection. From these 1260 patient records were reviewed. Appropriate eyecare was received by Australian patients at an average of 71% (95%CI 70%, 73%) of eligible encounters. The percentage of appropriateness of eyecare at the condition level for preventative, glaucoma and diabetic eyecare was 81% (95%CI 79%, 83%), 63% (95%CI 61%, 64%), and 69% (95%CI 66%, 73%), respectively. Appropriateness of eyecare delivery was lowest for the domains of history taking and physical examination for all eye conditions. There were pockets of excellence but consistent delivery of appropriate eyecare needs improvement, and gaps in eyecare delivery should be addressed.


Attenuated AMPA receptor expression allows glioblastoma cell survival in glutamate-rich environment.
Glioblastoma multiforme (GBM) cells secrete large amounts of glutamate that can trigger AMPA-type glutamate receptors (AMPARs). This commonly results in Na(+) and Ca(2+)-permeability and thereby in excitotoxic cell death of the surrounding neurons. Here we investigated how the GBM cells themselves survive in a glutamate-rich environment. In silico analysis of published reports shows down-regulation of all ionotropic glutamate receptors in GBM as compared to normal brain. In vitro, in all GBM samples tested, mRNA expression of AMPAR subunit GluR1, 2 and 4 was relatively low compared to adult and fetal total brain mRNA and adult cerebellum mRNA. These findings were in line with primary GBM samples, in which protein expression patterns were down-regulated as compared to the normal tissue. Furthermore, mislocalized expression of these receptors was found. Sequence analysis of GluR2 RNA in primary and established GBM cell lines showed that the GluR2 subunit was found to be partly unedited. Together with the lack of functional effect of AMPAR inhibition by NBQX our results suggest that down-regulation and afunctionality of AMPARs, enable GBM cells to survive in a high glutamate environment without going into excitotoxic cell death themselves. It can be speculated that specific AMPA receptor inhibitors may protect normal neurons against the high glutamate microenvironment of GBM tumors.


The reassembling process of the nonameric Mycobacterium tuberculosis small heat-shock protein Hsp16.3 occurs via a stepwise mechanism.
Conditions are reported under which the reassembled intermediates of the heat-shock protein Hsp16.3 after being denatured in 8 M urea were detected by mainly using urea-gradient PAGE (with modifications) and urea-denaturing pore-gradient PAGE. Hsp16.3 is the small heat-shock protein from Mycobacterium tuberculosis, which exists as a specific nonamer and was proposed to form a trimer-of-trimers structure. The refolding and reassembling of this protein was achieved rapidly by dilution or dialysis, suggesting an effectively spontaneous recovery of quaternary structure. Data presented in this report demonstrate that the in vitro reassembling process of Hsp16.3 protein occurs through a spontaneous and effective stepwise mechanism. Modified urea-gradient PAGE may provide a general method for studying the reassembling processes of other oligomeric proteins.


Characterization of mouse small intestinal cytochrome P450 expression.
The expression of biotransformation enzymes in mouse small intestine is poorly characterized, which limits the utility of transgenic or knockout mouse models for first-pass drug metabolism studies. In response, we have systematically examined the composition and inducibility of cytochrome P450 (P450) protein and mRNA in mouse small intestinal epithelial cells (enterocytes). RNA-PCR was conducted to confirm the expression and identity of CYP1A1, 1B1, 2B10, 2B19, 2B20, 2C29, 2C38, 2C40, 2E1, 3A11, 3A13, 3A16, 3A25, and 3A44 in the enterocytes of untreated mice, but CYP1A2, 2A4/5, 2A12, 2C37, 2C39, and 2F2 were not detected. The inducibility of CYP2B, 2C, and 3A subfamily forms was determined by real-time quantitative RNA-PCR. All five CYP3A forms were induced, in a range from 1.7- to 4.5-fold, by dexamethasone (DEX). Phenobarbital (PB) induced CYP2B9, CYP2B10, and CYP2B20 mRNAs and suppressed CYP2B19 mRNA levels. PB also induced CYP2C29 and CYP2C40, but not CYP2C38 mRNA. At the protein level, CYP1A1, CYP1B1, CYP2B, CYP2C, CYP2E1, and CYP3A were detected in enterocytes from untreated mice by immunoblot analysis. CYP1A1 was inducible by beta-naphthoflavone (BNF), CYP2B and CYP2C by PB, and CYP3A by DEX. CYP2B, 2C, and 3A proteins were all expressed at high levels proximally, and decreased distally. The inducibility of CYP1A1 followed a similar pattern. Intestinal P450 expression was compared between C57BL/6 (B6) and 129/sv (129) mice, strains commonly used in the preparation of transgenic and knockout mouse models. There was no significant strain difference in constitutive levels or induction patterns for CYP2B, 2C, and 3A protein. However, CYP1A1 was induced to a high level by BNF in B6 mice, but was not induced in the 129 mice.


One-stage correction of complex facial disproportion.
Patients with facial disharmony frequently have abnormal nasal form and disordered jaw relationships. Both orthognathic surgery and rhinoplasty are required to correct such facial disproportions. During a 10-year period 100 patients with a spectrum of indications have had concomitant orthognathic and rhinoplasty surgery by a team consisting of the same plastic surgeon, oral and maxillofacial surgeon, and orthodontist. Of this group 51 patients were operated on solely for aesthetic reasons, the majority being long face syndrome. All had a rhinoplasty usually with septal surgery. Depending on the deformity, the jaw surgery varied: 5 patients had mandibular surgery only, 12 had maxillary surgery alone, and the remaining 34 patients had both mandibular and maxillary procedures. Patients were followed for between 1 and 62 months by the plastic surgeon and for at least 2 years by the orthodontist. There was no orthognathic relapse or other major complications, but 4 patients required secondary minor nasal tip surgery under local anesthesia and 2 patients had persisting unilateral inferior alveolar nerve damage. Orthognathic surgery and rhinoplasty are not routinely performed concomitantly due to the difficulty in predicting the outcome of the soft-tissue relationships and increased morbidity. In this series, a one-stage approach was used to provide facial harmony. This reduces the overall surgical and anesthetic morbidity, inconvenience, and expense, and has resulted in good cosmetic and functional results. Therefore, it is suggested that with a competent team, orthognathic surgery and rhinoplasty can be performed concomitantly with dependable results and without significant complications.


The use of radioactive iodine-125 seed localization in patients with non-palpable breast cancer: a comparison with the radioguided occult lesion localization with 99m technetium.
Radioactive Seed Localization with a radioactive iodine-125 seed (RSL) and Radioguided Occult Lesion Localization with 99mTechnetium colloid (ROLL) are both attractive alternatives to wire localization for guiding breast conserving surgery (BCS) of non-palpable breast cancer. The aim of this study was to evaluate and compare the efficacy of RSL and ROLL. We retrospectively analyzed 387 patients with unifocal non-palpable ductal carcinoma in situ (DCIS) or invasive carcinoma treated with BCS at the Netherlands Cancer Institute. In total 403 non-palpable lesions were localized either by RSL (N = 128) or by ROLL (N = 275). Primary outcome measures were positive margins and re-excision rates; the secondary outcome measure was weight of the specimen. Pre-operative mammography or ultrasound showed similar sizes of DCIS and invasive tumours in both RSL and ROLL groups. In the RSL group, more lesions were DCIS (58%) than in the ROLL group, where 32% of the lesions were pure DCIS. The proportions of focally positive margins (11% vs. 10%) and more than focally positive margins (9% vs. 9%) were comparable between the RSL and the ROLL group, resulting in the same re-excision rate in both RSL and ROLL groups (9% vs. 10%). For DCIS lesions, the specimen weight was significantly lower in the RSL group than in the ROLL group after adjusting for tumour size on mammography (12 g; 95% CI 2.6-21). Margin status and re-excision rates were comparable for RSL and ROLL in patients with non-palpable breast lesions. Because of the significant lower weight of the resected specimen in DCIS, the feasibility of position verification of the I-125 seed and more convenient logistics, we favour RSL over ROLL to guide breast-conserving therapy.


The portrait of Dr Edward Harrison MD (1766-1838).
The portrait of the London and Horncastle Physician Edward Harrison, painted and displayed in 1823, was editorially criticized by one of the medical journals of the time. After Harrison died the portrait remained in the family estate until 1938, after which it was presented to the National Gallery of Ireland. The image of Dr Harrison had never been displayed in any of his medical writings, nor in any other medical historical works, until 2008. This paper provides some history of the criticism of the portrait, similar to the historical vignette of the portraiture of William Harvey and outlines the detective work to track down the location of the portrait.


Toxoplasma gondii profilin promotes recruitment of Ly6Chi CCR2+ inflammatory monocytes that can confer resistance to bacterial infection.
Ly6C+ inflammatory monocytes are essential to host defense against Toxoplasma gondii, Listeria monocytogenes and other infections. During T. gondii infection impaired inflammatory monocyte emigration results in severe inflammation and failure to control parasite replication. However, the T. gondii factors that elicit these monocytes are unknown. Early studies from the Remington laboratory showed that mice with a chronic T. gondii infection survive lethal co-infections with unrelated pathogens, including L. monocytogenes, but a mechanistic analysis was not performed. Here we report that this enhanced survival against L. monocytogenes is due to early reduction of bacterial burdens and elicitation of Ly6C+ inflammatory monocytes. We demonstrate that a single TLR11/TLR12 ligand profilin (TgPRF) was sufficient to reduce bacterial burdens similar to T. gondii chronic infection. Stimulation with TgPRF was also sufficient to enhance animal survival when administered either pre- or post-Listeria infection. The ability of TgPRF to reduce L. monocytogenes burdens was dependent on TLR11 and required IFN-γ but was not dependent on IL-12 signaling. TgPRF induced rapid production of MCP-1 and resulted in trafficking of Ly6Chi CCR2+ inflammatory monocytes and Ly6G+ neutrophils into the blood and spleen. Stimulation with TgPRF reduced L. monocytogenes burdens in mice depleted with the Ly6G specific MAb 1A8, but not in Ly6C/Ly6G specific RB6-8C5 depleted or CCR2-/- mice, indicating that only inflammatory monocytes are required for TgPRF-induced reduction in bacterial burdens. These results demonstrate that stimulation of TLR11 by TgPRF is a mechanism to promote the emigration of Ly6Chi CCR2+ monocytes, and that TgPRF recruited inflammatory monocytes can provide an immunological benefit against an unrelated pathogen.


Synthesis of oxygen-linked 8-phenoxyl-deoxyguanosine nucleoside analogues.
Nucleobase adducts, which form in vivo by the nucleophilic attack of nucleobases on exogenous electrophilic species, can impact conformation and biological influences of the adducted nucleoside. Contemporary studies aim to address the occurrence and relevance of O-linked 8-phenoxy-purine adducts; however, preparative techniques for synthesizing these nucleosides were not previously described. Reported herein is a relatively facile synthesis of O-linked 8-dG phenol adducts with a wide variety of electron-donating, electron-withdrawing, and sterically demanding phenols.


Structure of anticancer ruthenium half-sandwich complex bound to glycogen synthase kinase 3β.
The 3.15-Å-resolution crystal structure of the R enantiomer of the highly bioactive and antiproliferative half-sandwich ruthenium complex DW12 bound to the ATP binding site of glycogen synthase kinase 3β (GSK-3β) is reported and the binding is compared with the GSK-3β binding of staurosporine and other organic inhibitors. The structure reveals a close packing of the organometallic inhibitor in the ATP binding site of GSK-3β via an induced-fit mechanism. The molecular structure of (R)-DW12 with the CO ligand oriented perpendicular to the pyridocarbazole heterocycle allows the complex to stretch the whole distance sandwiched between the faces of the N- and C-terminal lobes and to interact tightly with the flexible glycine-rich loop, which is uncommon for the interaction of GSK-3β with organic inhibitors.


MR-guided radiofrequency ablation of hepatocellular carcinoma: long-term effectiveness.
To evaluate long-term effectiveness of magnetic resonance (MR)-guided radiofrequency (RF) ablation of hepatocellular carcinoma (HCC). This prospective study was approved by the institutional review board. In 20 patients, 28 HCCs (mean diameter, 28.0 mm; range, 6-58 mm) were treated with 25 sessions of MR-guided RF ablation. Previous chemoembolization had been performed in nine HCCs with diameters greater than 3 cm. The entire RF ablation procedures were carried out on a 0.2-T open MR system. Placement of MR-compatible internally cooled electrodes was performed under MR fluoroscopic imaging with fast gradient-echo sequences. Therapeutic assessment was based on dynamic MR-imaging (1.5 T) at a mean follow-up of 24.2 months (range, 6-52 mo). MR-guided RF ablation was technically successful in all 25 sessions (100%), as assessed at the end of each session. T2-weighted sequences were accurate to monitor the ablation zone and supported guidance of overlapping ablations if necessary. Technique effectiveness, defined as complete ablation confirmed at MR imaging 4 months after RF ablation, was achieved in 27 of 28 HCCs (96.4%). To achieve complete ablation, 25 of 27 tumors (92.6%) were treated in a single session and two tumors were treated twice. In one tumor initially defined as having been treated with technically effective RF ablation, local tumor progression was detected more than 4 months after ablation. Consequently, the available follow-up indicated complete ablation in 26 of 28 HCCs (92.9%). There was one major complication (4.0%) and one minor complication (4.0%). On a long-term basis, MR-guided RF ablation is an effective therapy option in the treatment of HCC.


Surface- and tip-enhanced Raman scattering of bradykinin onto the colloidal suspended Ag surface.
In this paper, surface- (SERS) and tip-enhanced Raman scattering (TERS) techniques were used to determine the adsorption mode of bradykinin (BK), a small peptide implicated in, for example, carcinoma growth, onto colloidal suspended Ag surfaces under various environmental conditions, including: peptide concentrations (10(-5)-10(-7) M), excitation wavelengths (514.5 and 785.0 nm), and pH of aqueous sol solutions (from pH = 3 to pH = 11). The metal surface plasmon and rheology of the colloidal suspended Ag surface were explored by ultraviolet-visible (UV-Vis) spectroscopy and atomic force/scanning electron microscopy (AFM/SEM). The SERS results indicated that the peptide concentration of 10(-5) M was the optimal peptide concentration for monolayer colloidal coverage. The Phe(5/8) and Arg(9) residues of BK generally participated in the interactions with colloidal suspended Ag surfaces. The amide group appeared to be arranged in the same manner to the Ag surface in the pH range of 3 to 11. At acidic pH of the solution (pH = 3 to 5), the BK -COO(-) terminal group binds to the Ag surface as a bidentate (at pH = 3) or monodentate (at pH = 5) chelating ligand. At pH = 11, the imino group of Arg(9), probably due to its -C[double bond, length as m-dash]N(⊕)H2 protonation state, was not involved in the interaction with Ag. The reduction in the solution alkalinity (pH = 9) produced the deprotonation of the -C=N(⊕)H2 group followed by group rearrangement in a way favoring the interaction between the lone electron pair on N and Ag. The TERS studies confirmed the proposed, on the basis of SERS, behavior of BK onto the colloidal suspended Ag at pH = 7 and showed that in different points of the colloidal suspended Ag surface the same peptide fragments approximately having the same orientations with respect to this surface interact with it.


Optimisation methodologies and algorithms for research on catalysis employing high-throughput methods: comparison using the Selox benchmark.
The Selox is a catalytic benchmark for the selective CO oxidation reaction in the presence of H(2), in the form of mathematical equations obtained via modelling of experimental results. The optimisation efficiencies of several Global Optimisation algorithms were studied using the Selox benchmark. Genetic Algorithms, Evolutionary Strategies, Simulated Annealing, Taboo Search and Genetic Algorithms hybridised with Knowledge Discovery procedures were the methods compared. A Design of Experiments search strategy was also exemplified using this benchmark. The main differences regarding the applicability of DoE and Global optimisation techniques are highlighted. Evolutionary strategies, Genetic algorithms, using the sharing procedure, and the Hybrid Genetic algorithms proved to be the most successful in the benchmark optimisation.


Interchange between collagenous and lymphocytic colitis in severe disease with autoimmune associations requiring colectomy: a case report.
Collagenous colitis and lymphocytic colitis present with a similar clinical picture. Whether these conditions are separate entities or whether they represent different pathological stages of the same condition is an unresolved issue. This is a case of collagenous colitis following a fulminant course in which a colectomy was necessary. In the operative specimen the thickened collagen plate, which had been present only two weeks preoperatively had been lost and the pathology was of a lymphocytic colitis. Six months postoperatively this patient developed a CREST syndrome and primary biliary cirrhosis. This case shows the lability of the collagen plate and the common ground between collagenous and lymphocytic colitis, and presents evidence that these two conditions are different manifestations of the same disease. It also describes for the first time an association between collagenous colitis and CREST syndrome and primary biliary cirrhosis.


A new approach in muscle training to rehabilitate the hand in leprosy.
Rehabilitation of the hand in Leprosy with its deformities and anaesthesia no doubt poses a formidable challenge as far as rehabilitation is concerned. Here we have applied a combination of latest concepts in sports physiology and the exercise followed by the students of ancient Indian martial art known as Krishni Vidya. This exercise consists of crumpling a newspaper sheet with a single hand without any external support. We are unable to explain the type of exercise done by the people before existence of newspaper. They might be using naturally occurring materials for the same like woodbark. With this exercise we have been able to improve the function of the hand remarkably well and the rehabilitation time following surgery is drastically reduced, with function returning close to normalcy. The full function is achieved within four weeks from the date of removal of plaster as compared to eight to ten weeks or more with conventional physical therapy. This full function is not only in mobility but also in co-ordination, static and dynamic strength, speed of movement and flexibility required of a normal hand. 16 patients operated for lumbrical replacement (using sublimis as a motor from the long finger) at the Dr. Bandorawalla Leprosy Hospital were put under the paper crumpling exercise. It was observed that the average time for the achievement of fully close fist was two weeks. (All of them had thumb function intact, i.e. either normal or operated-out of sixteen, four patients were operated for opponens prior to lumbrical replacement).


Structural insight into DNA binding and oligomerization of the multifunctional Cox protein of bacteriophage P2.
The Cox protein from bacteriophage P2 is a small multifunctional DNA-binding protein. It is involved in site-specific recombination leading to P2 prophage excision and functions as a transcriptional repressor of the P2 Pc promoter. Furthermore, it transcriptionally activates the unrelated, defective prophage P4 that depends on phage P2 late gene products for lytic growth. In this article, we have investigated the structural determinants to understand how P2 Cox performs these different functions. We have solved the structure of P2 Cox to 2.4 Å resolution. Interestingly, P2 Cox crystallized in a continuous oligomeric spiral with its DNA-binding helix and wing positioned outwards. The extended C-terminal part of P2 Cox is largely responsible for the oligomerization in the structure. The spacing between the repeating DNA-binding elements along the helical P2 Cox filament is consistent with DNA binding along the filament. Functional analyses of alanine mutants in P2 Cox argue for the importance of key residues for protein function. We here present the first structure from the Cox protein family and, together with previous biochemical observations, propose that P2 Cox achieves its various functions by specific binding of DNA while wrapping the DNA around its helical oligomer.


Effects of transgenic overproduction of CRH on anxiety-like behaviour.
Central administration of corticotropin-releasing hormone increases anxiety-like behaviour and arousal in rodents, and increased anxiety-like behaviour has been shown in mice overproducing corticotropin-releasing hormone on an elevated plus maze and in a dark-light emergence task. However, evidence is accumulating that measures obtained from different anxiety tasks may reflect different aspects of anxiety-like behaviour in animals. We therefore tested mice overproducing corticotropin-releasing hormone in a battery of paradigms, studying spontaneous behaviour after a mild stressor, tasks of innate anxiety-like behaviour (light-dark box), lick suppression (Vogel conflict), conditioned fear, and forced swimming. Exploratory behaviour was studied in a 16-hole board task. Furthermore, pain threshold, water intake, locomotor activity and sensorimotor learning/co-ordination were tested to control for confounding factors. In line with previous findings, increased anxiety-like behaviour of transgenic mice was observed in the light-dark box paradigm. However, no differences were seen in the conflict paradigm. Conditioned fear was decreased 1 h but not 24 h after conditioning in transgenic mice, and immobility was decreased [corrected] in forced swimming in corticotropin-releasing hormone overexpressors. Locomotor activity in a novel open field and on the hole board was reduced in transgenics. Exploratory behaviour (hole pokes) was reduced during initial exploration of an unfamiliar hole board. Moreover, sensorimotor performance on a rotorod was impaired, and water intake was reduced in corticotropin-releasing hormone overproducing mice, while no changes were seen in nociception. No differences in locomotor activity were seen in a second group of mice, tested in a familiar open field. When these animals were challenged with diazepam, transgenic mice were less susceptible to the sedative effects of the drug on locomotor activity. These data suggest that corticotropin-releasing hormone overproduction leads to specific effects in a subset of anxiety paradigms, and that these transgenic mice suffer from a motor deficit in addition to altered anxiety-like behaviour/arousal.


Discovery of (S)-3-(4-(benzyloxy)phenyl)-2-(2-phenoxyacetamido)propanoic acid derivatives as a new class of GPR34 antagonists.
GPR34 is a rhodopsin-like class G protein-coupled receptor (GPCR) that is involved in the development and progression of several diseases. Despite its importance, effective targeting strategies are lacking. We herein report a series of (S)-3-(4-(benzyloxy)phenyl)-2-(2-phenoxyacetamido)propanoic acid derivatives as a new class of GPR34 antagonists. Structure-activity relationship (SAR) studies led to the identification of the most potent compound, 5e, which displayed an IC50 value of 0.680 μM in the GloSensor cAMP assay and 0.059 μM in the Tango assay. 5e demonstrated low cytotoxicity and high selectivity in vitro, and it was able to dose-dependently inhibit Lysophosphatidylserine-induced ERK1/2 phosphorylation in CHO cells expressing GPR34. Furthermore, 5e displayed excellent efficacy in a mouse model of neuropathic pain without any apparent signs of toxicity. Collectively, this study has identified a promising compound, which shows great potential in the development of potent antagonists with a new chemical scaffold targeting GPR34.


Percutaneous balloon valvotomy in tricuspid stenosis.
This report describes the first successful treatment of tricuspid stenosis by percutaneous double balloon valvotomy. There was a dramatic reduction of the tricuspid valve gradient, with an increase in calculated valve area, together with an increase in resting cardiac output and symptomatic relief. The feasibility of the non-surgical treatment of severe tricuspid stenosis was demonstrated unequivocally.


Adverse effects of outdoor pollution in the elderly.
With fewer newborns and people living longer, older people are making up an increasing fraction of the total population. Epidemiological evidence shows that older-age-related health problems affect a wide and expanding proportion of the world population. One of the major epidemiological trends of this century is the rise of chronic diseases that affect more elderly than younger people. A total of 3.7 million premature deaths worldwide in 2012 are attributable to outdoor air pollution; the susceptibility to adverse effects of air pollution is expected to differ widely between people and within the same person, and also over time. Frailty history, a measure of multi-system decline, modifies cumulative associations between air pollution and lung function. Moreover, pre-existing diseases may determine susceptibility. In the elderly, due to comorbidity, exposure to air pollutants may even be fatal. Rapid and not-well-planned urbanization is associated with high level of ambient air pollution, mainly caused by vehicular exhausts. In general, there is sufficient evidence of the adverse effects related to short-term exposure, while fewer studies have addressed the longer-term health effects. Increased pollution exposures have been associated with increased mortality, hospital admissions/emergency-room visits, mainly due to exacerbations of chronic diseases or to respiratory tract infections (e.g., pneumonia). These effects may also be modulated by ambient temperature and many studies show that the elderly are mostly vulnerable to heat waves. The association between heat and mortality in the elderly is well-documented, while less is known regarding the associations with hospital admissions. Chronic exposure to elevated levels of air pollution has been related to the incidence of chronic obstructive pulmonary disease (COPD), chronic bronchitis (CB), asthma, and emphysema. There is also growing evidence suggesting adverse effects on lung function related to long-term exposure to ambient air pollution. Few studies have assessed long-term mortality in the elderly. It is still unclear what are the pollutants most damaging to the health of the elderly. It seems that elderly subjects are more vulnerable to particulate matter (PM) than to other pollutants, with particular effect on daily cardio-respiratory mortality and acute hospital admissions. Not many studies have targeted elderly people specifically, as well as specific respiratory morbidity. Most data have shown higher risks in the elderly compared to the rest of the population. Future epidemiological cohort studies need to keep investigating the health effects of air pollutants (mainly cardiopulmonary diseases) on the elderly.


Splenic recurrence of liver hydatid cyst and spleen preserving therapy.
Hydatid cyst disease remains a considerable public health problem, especially in pastoral and farming regions. Although the spleen is the third most commonly affected organ after the liver and lungs, splenic hydatid cyst is an uncommon entity even in areas that are endemic for echinococcosis. The recurrence rate after surgical therapy of the liver hydatid cyst is reported as 6.8-22.3 percent. Recurrences most frequently occur in the liver. Extrahepatic recurrences occur in the lung or peritoneum and the serosa of the abdominal organs. Splenic recurrence of liver hydatid cyst has not previously been reported. The most common surgical therapy is splenectomy, and the other option is spleen preserving surgery. We report the first case of recurrent splenic hydatid cyst in the spleen and liver synchronously after surgical therapy for liver hydatid disease. The patient was treated with liver resection and spleen preserving surgery.


Adenovirus 14p1 Immunopathogenesis during Lung Infection in the Syrian Hamster.
Adenovirus (Ad) infections are usually mild and self-limited, with minimal inflammatory responses. During worldwide outbreaks, Ad14p1, an emerging Ad14 variant, has caused severe pulmonary disease, including acute respiratory distress syndrome (ARDS). This increased pathogenicity of Ad14p1 is not completely understood. In initial studies, we observed that infection of Syrian hamsters with Ad14p1 can cause a patchy bronchopneumonia, with an increased intensity of inflammation, compared to wild type Ad14 infection. The current study compared the dynamics of the immunopathogenesis of Ad14 and Ad14p1 infection of hamster lungs through the first two weeks after infection. Little difference was seen in infection-induced inflammation at day 1. Beginning at day 3, Ad14p1-infected hamsters showed marked inflammation that continued through to day 7. The inflammation began to resolve by day 10 but was still detectable at day 14. In contrast, Ad14-infected hamsters showed little inflammation during the 14-day period of observation. Inflammatory cell type analysis revealed that, at day 1, hamsters infected with either virus had predominantly neutrophil infiltration that began to resolve by day 3. However, at day 5, Ad14p1-infected hamsters had a second wave of neutrophil infiltration that was accompanied by edema which persisted to a variable extent through to day 10. These differences were not explained by an increased Ad14p1 replication rate, compared with Ad14 in vitro, but there was prolonged persistence of Ad14p1 in hamster lungs. There were differences in lung tissue cytokine and chemokine responses to Ad14p1 vs. Ad14 infection that might account for the increased leukocyte infiltrates in Ad14p1-infected hamsters. This animal model characterization provides the basis for future translational studies of the viral genetic mechanisms that control the increased immunopathogenesis of the emergent, Ad14p1 strain.


Transcellular metabolism of eicosanoids.
In this chapter, current concepts of eicosanoid biochemistry and function have been summarized. Emphasis has been placed on the importance of ascertaining a complete profile of eicosanoids synthesized by a given tissue under varying conditions of stimulation. The concept of transient intermediates, which are involved in each of the known pathways, was reviewed. Such intermediates can also represent substrate for cell-cell interactions, which have been classified and discussed. Knowledge of the synthetic mechanisms, metabolism, and catabolism of eicosanoids has surpassed our comprehension of their biologic actions. It can be speculated that eicosanoids reinforce or synergize normal homeostatic mechanisms that might proceed less effectively in their absence but occur more efficiently when they are produced. Incomplete comprehension of the functional role of eicosanoids has retarded the development of definitive pharmaceutical approaches toward inhibition or stimulation of eicosanoid production in pathophysiologic situations. With regard to thrombosis, the inflammatory response and host-defense mechanisms, leukotrienes, which have been shown to act on vascular endothelium and smooth muscle, may play an important role in occlusive vascular disease. Since leukotrienes and other hydroxy acids such as 12-HETE are not inhibited by aspirin or nonsteroidal anti-inflammatory agents, development of a unifying concept is difficult. It appears that we are approaching a point in time for reevaluation and reassessment of the role of the eicosanoid pathway in hemostasis and thrombosis. Most importantly, recent eicosanoid research has provided answers to biologic phenomena that were not explicable in the past and also explains why therapeutic trials in occlusive vascular disease were not as successful as predicted on theoretical grounds.


Individual differences in mood reactions to d-amphetamine: a test of three personality factors.
Individual differences in self-reported mood following either 5 mg or 10 mg d-amphetamine challenge were examined in order to test the modifying role of three factors of personality, viz., the Eysencks' psychoticism, Cloninger's novelty seeking, and Depue and Collins' extraversion. In a double-blind study, mood measures (energetic arousal, tense arousal, and hedonic tone) were taken immediately following a single-dose of d-amphetamine and then again after 90 min. The results showed significant psychoticism x d-amphetamine interactions for both drug doses: d-amphetamine increased energetic arousal and hedonic tone, and reduced tense arousal, only in low psychoticism individuals; in high psychoticism individuals, it led to lowered energetic arousal and hedonic tone, and increased tense arousal. Neither novelty seeking nor extraversion modified the effects of d-amphetamine. These data suggest a link between psychoticism and dopaminergic functioning, although they do not rule out the involvement of other transmitter systems (e.g. noradrenergic). In common with other studies, such findings point to the important role that well-established factors of personality play in accounting for individual differences in reactions to psychoactive drugs. It is concluded that the routine inclusion of personality measures in future psychopharmacological studies may help to refine the characterization of drug effects.


Selection history: How reward modulates selectivity of visual attention.
Visual attention enables us to selectively prioritize or suppress information in the environment. Prominent models concerned with the control of visual attention differentiate between goal-directed, top-down and stimulus-driven, bottom-up control, with the former determined by current selection goals and the latter determined by physical salience. In the current review, we discuss recent studies that demonstrate that attentional selection does not need to be the result of top-down or bottom-up processing but, instead, is often driven by lingering biases due to the "history" of former attention deployments. This review mainly focuses on reward-based history effects; yet other types of history effects such as (intertrial) priming, statistical learning and affective conditioning are also discussed. We argue that evidence from behavioral, eye-movement and neuroimaging studies supports the idea that selection history modulates the topographical landscape of spatial "priority" maps, such that attention is biased toward locations having the highest activation on this map.


Biological and molecular characterization of lettuce mosaic virus from Tehran province in Iran.
In this study, lettuce samples having LMV infection symptoms were collected from Tehran fields during 2003. Samples tested for LMV infection by immuno printing. Three positive samples in immuno printing collected and their characteristics were determined. In mechanical inoculation, these Isolates produced symptoms on Chenopodium quinoa, C. amaranticolor, Gomphrena globosa, Nicotiana benthamiana, Lactuca sativa cv. Mantilia and cv. Terocadero (which contains the mol1 resistance gene and susceptible respectively), but not cv. Salinas 88 (which contains the mol2 resistance gene). LMV was purified and LMV polyclonal antiserum was produced in rabbit by a series of intravenous and intramuscular injections, the precipitin titre of this antiserum was 1:1024. Gel double diffusion test (GDDT) was performed, and precipitin bands appeared. SDS-PAGE and western blotting showed the presence of coat protein 29 kDa. In IC-RT-PCR with on LMV specific primer pair, an approximately 1300 bp fragment was amplified.


The role of laser-induced autofluorescence spectroscopy in bladder tumor detection. Dependence on the excitation wavelength.
We assessed the ability of laser-induced autofluorescence spectroscopy to distinguish neoplastic urothelial bladder lesions from normal or nonspecific inflammatory mucosa. Three different pulsed laser excitation wavelengths were used successively: 308 nm (xenium chloride excimer laser), 337 nm (nitrogen laser) and 480 nm (coumarin dye laser). The excitation light was delivered by a specially devised multifiber catheter connected to a 1-mm core diameter silica monofiber introduced through the working channel of a standard cystoscope with saline irrigation. The captured fluorescence light was focused onto an optical multichannel analyzer detection system. Performance of this device was evaluated in 25 patients after obtaining consent and immediately before transurethral resection of a bladder tumor. Spectroscopic results were compared with histological findings. At 337- and 480-nm excitation wavelengths, the overall fluorescence intensity of bladder tumors was clearly decreased compared to normal urothelial mucosa regardless of tumor stage and grade. At the 308-nm excitation wavelength, the shape of the tumor spectra, including carcinoma in situ, was markedly different from that of normal or nonspecific inflammatory mucosa. No absolute intensity determinations were required in this situation, since a definite diagnosis could be established based on the fluorescence intensity ratio at 360 and 440 nm. This spectroscopic study could be particularly useful in designing a simplified autofluorescence imaging device for detection of occult urothelial neoplasms.


Emerging roles of HECT-type E3 ubiquitin ligases in autophagy regulation.
Autophagy is a conserved self-eating process that delivers cytoplasmic material to the lysosome to allow degradation of intracellular components, including soluble, unfolded and aggregated proteins, damaged organelles, and invading microorganisms. Autophagy provides a homeostatic control mechanism and is essential for balancing sources of energy in response to nutrient stress. Autophagic dysfunction or dysregulation has been implicated in several human pathologies, including cancer and neurodegeneration, and its modulation has substantial potential as a therapeutic strategy. Given the relevant clinical and therapeutic implications of autophagy, there is emerging intense interest in the identification of the key factors regulating the components of the autophagic machinery. Various post-translational modifications, including ubiquitylation, have been implicated in autophagy control. The list of the E3 ubiquitin protein ligases involved in the regulation of several steps of the autophagic process is continuously growing. In this review, we will focus on recent advances in the understanding of the role of the homologous to the E6AP carboxyl terminus-type E3 ubiquitin ligases in autophagy control.


On-chip screening method for cell migration genes based on a transfection microarray.
Cell migration plays a major role in a variety of biological processes and a detailed understanding of associated mechanisms should lead to advances in the medical sciences, for example, in drug discovery for cancer therapy. However, the traditional methods used for analysis of cell migration cannot easily be scaled up for high-throughput screening. In this study, we have attempted to develop a novel simple method for high-throughput phenotypic screening for the identification of genes that are required for cell migration. As the appropriate cell line for the method, we found NBT-L2b cells that would be suitable for screening of migration-related genes in our method without influence by other cellular processes. Moreover, the idea for printing both the labeled fibronectin, for identification of the starting region of a cell, and the green fluorescent protein (GFP) expression vector, for identification of cells that had been transfected with siRNA and of the end point of migration, brings a rapid and efficient high-throughput screening procedure. Our new method will lead to an enhanced understanding of cell migration.


[A less known cause of incoercible vomiting].
Cannabis is a widely spread drug which chronic abuse may cause diverse adverse effects on the body, in the short and long term. Among these effects, we can cite digestive disorders, mainly characterized by the onset of cyclical vomiting illness. Vomiting symptoms can be temporarily relieved by hot water showers. As a matter of fact, many patients adopt a compulsive bathing behavior characterized by the irrepressible need of having showers or baths at a high temperature. We describe the case of two patients (22 and 23 years-old) who consume cannabis on a regular basis. They were admitted several times to hospital emergency rooms for complaints about uncontrollable vomiting with recurring abdominal pain. Multiple investigations and hospitalizations did not determine a precise diagnosis explaining the symptomatology. The final diagnosis is the vomiting syndrome caused by the consumption of cannabis.


Dietary Intake, Nutritional Adequacy and Food Sources of Total Fat and Fatty Acids, and Relationships with Personal and Family Factors in Spanish Children Aged One to <10 Years: Results of the EsNuPI Study.
We aimed to determine the usual intake of total fat, fatty acids (FAs), and their main food sources in a representative cohort of the Spanish pediatric population aged 1 to <10 years (n = 707) who consumed all types of milk and an age-matched cohort who consumed adapted milk over the last year (including follow-on formula, toddler's milk, growing-up milk, and fortified and enriched milks) (n = 741) who were participants in the EsNuPI study (in English, Nutritional Study in the Spanish Pediatric Population). Dietary intake, measured through two 24 h dietary recalls, was compared to the European Food Safety Authority (EFSA) and the Food and Agriculture Organization of the United Nations (UN-FAO) recommendations. Both cohorts showed a high intake of saturated fatty acids (SFAs), according to FAO recommendations, as there are no numerical recommendations for SFAs at EFSA. Also, low intake of essential fatty acids (EFAs; linoleic acid (LA) and α-linolenic acid (ALA)) and long-chain polyunsaturated fatty acids (LC-PUFA) of the n-3 series, mainly docosahexaenoic acid (DHA) were observed according to EFSA and FAO recommendations. The three main sources of total fat and different FAs were milk and dairy products, oils and fats, and meat and meat products. The consumption of adapted milk was one of the main factors associated with better adherence to the nutritional recommendations of total fat, SFAs, EFAs, PUFAs; and resulted as the main factor associated with better adherence to n-3 fatty acids intake recommendations. Knowledge of the dietary intake and food sources of total fat and FAs in children could help in designing and promoting effective and practical age-targeted guidelines to promote the consumption of EFA- and n-3 PUFA-rich foods in this stage of life.


Complications of percutaneous tracheostomy.
Percutaneous tracheostomy is a technique that, reputedly, is simple to perform and causes few complications. It is routinely used in intensive care. We present two patients with tracheal stenosis. In one patient, we had to perform an anastomotic resection to cure the patient; in the other patient, we had to place an endoluminal conformer. To our knowledge, this complication has not been reported in association with the use of this technique.


Risk factors for intraoperative hypertensive crisis in patients with pheochromocytomas and sympathetic paragangliomas.
To identify presurgical and surgical risk factors for intraoperative hypertensive crisis in patients with pheochromocytomas and sympathetic paragangliomas (PGLs) (PPGLs). Retrospective multicenter cohort study of patients with PPGLs from 18 tertiary hospitals. Intraoperative hypertensive crisis was defined as systolic blood pressure (SBP) greater than 200 mmHg lasting more than 1 min and postoperative hypertensive crisis as SBP greater than 180 mmHg or diastolic blood pressure (DBP) greater than 110 mmHg. A total of 296 surgeries were included. Alpha presurgical blockade was employed in 93.2% of the cases and beta-adrenergic in 53.4%. Hypertensive crisis occurred in 20.3% (n = 60) of the surgeries: intraoperative crisis in 56 and/or postoperative crisis in 6 cases (2 cases had both types of crises). We identified as risk factors of intraoperative hypertensive crisis, absence of presurgical glucocorticoid therapy (odds ratio [OR] 3.48; 95% confidence interval [CI] 1.19-10.12) higher presurgical SBP (OR 1.22 per each 10 mmHg, 95% CI 1.03-1.45), a larger tumor size (OR 1.09 per each 10 mm, 95% CI 1.00-1.19) and absence of oral sodium repletion (OR 2.59, 95% CI 1.25-5.35). Patients with hypertensive crisis had a higher rate of intraoperative bleeding (P < 0.001), of intraoperative hemodynamic instability (P < 0.001) and of intraoperative hypotensive episodes (P < 0.001) than those without hypertensive crisis. Intraoperative hypertensive crisis occurs in up to 20% of the PPGL resections. Patients not pretreated with glucocorticoid therapy before surgery, with larger tumors and higher presurgical SBP and who do not receive oral sodium repletion had a higher risk for developing hypertensive crisis during and after PPGL surgery.


Proton therapy dosimetry using positron emission tomography.
Protons deposit most of their kinetic energy at the end of their path with no energy deposition beyond the range, making proton therapy a valuable option for treating tumors while sparing surrounding tissues. It is imperative to know the location of the dose deposition to ensure the tumor, and not healthy tissue, is being irradiated. To be able to extract this information in a clinical situation, an accurate dosimetry measurement system is required. There are currently two in vivo methods that are being used for proton therapy dosimetry: (1) online or in-beam monitoring and (2) offline monitoring, both using positron emission tomography (PET) systems. The theory behind using PET is that protons experience inelastic collisions with atoms in tissues resulting in nuclear reactions creating positron emitters. By acquiring a PET image following treatment, the location of the positron emitters in the patient, and therefore the path of the proton beam, can be determined. Coupling the information from the PET image with the patient's anatomy, it is possible to monitor the location of the tumor and the location of the dose deposition. This review summarizes current research investigating both of these methods with promising results and reviews the limitations along with the advantages of each method.


Extracellular signal-regulated kinase and AP-1 pathways are involved in reactive oxygen species-induced urokinase plasminogen activator receptor expression in human gastric cancer cells.
Overexpression of urokinase plasminogen activator receptor (uPAR) is known to correlate closely with tumor cell invasion and metastasis. In gastric cancer, however, the mechanism for induction of uPAR remains to be elucidated. In this study, to investigate the effect of reactive oxygen species (ROS) on uPAR expression and the underlying signal pathways in human gastric cancer AGS cells, phenazine methosulfate (PMS) and H2O2 were used as ROS generator. PMS and H2O2 induced the uPAR expression in time- and concentration-dependent manner. PMS and H2O2 also induced the activation of extracellular signal-regulated kinases (Erk)-1/2. A specific inhibitor of MEK-1 (PD980590) was found to suppress the PMS-induced uPAR expression and the uPAR promoter activity. Expression of vectors encoding a mutated-type MEK-1 resulted in decreases in the uPAR promoter activity. Electrophoretic mobility shift assay revealed that PMS increased time-dependently the DNA binding activity of AP-1. Transient transfection studies using AP-1 decoy confirmed that the activation of AP-1 is involved in PMS-induced uPAR upregulation. The AGS cells pretreated with PMS showed a remarkably enhanced invasiveness and this effect was partially abrogated by uPAR neutralizing antibodies. The above results suggest that ROS induces uPAR expression via Erk-1/2 and AP-1 signaling pathways and, in turn, stimulates the cell invasiveness in human gastric cancer AGS cells.


How agarose gels surrounding PLGA implants limit swelling and slow down drug release.
The aim of this study was to better understand to which extent and in which way the presence of an agarose gel (mimicking living tissue) around a PLGA [poly(lactic-co-glycolic acid)] implant affects the resulting drug release kinetics. Ibuprofen-loaded implants were prepared by hot melt extrusion. Drug release was measured upon exposure to phosphate buffer pH 7.4 in Eppendorf tubes, as well as upon inclusion into an agarose gel which was exposed to phosphate buffer pH 7.4 in an Eppendorf tube or in a transwell plate. Dynamic changes in the implants' dry & wet mass and dimensions were monitored gravimetrically and by optical macroscopy. Implant erosion and polymer degradation were observed by SEM and GPC. Different pH indicators were used to measure pH changes in the bulk fluids, gels and within the implants during drug release. Ibuprofen release was bi-phasic in all cases: A zero order release phase (~20% of the dose) was followed by a more rapid, final drug release phase. Interestingly, the presence of the hydrogel delayed the onset of the 2nd release phase. This could be attributed to the sterical hindrance of implant swelling: After a certain lag time, the degrading PLGA matrix becomes sufficiently hydrophilic and mechanically instable to allow for the penetration of substantial amounts of water into the system. This fundamentally changes the conditions for drug release: The latter becomes much more mobile and is more rapidly released. A gel surrounding the implant mechanically hinders system swelling and, thus, slows down drug release. These observations also strengthen the hypothesis of the "orchestrating" role of PLGA swelling for the control of drug release and can help developing more realistic in vitro release set-ups.


Soft ticks perform evaporative cooling during blood-feeding.
Feeding on the blood of warm-blooded vertebrates is associated to thermal stress in haematophagous arthropods. It has been demonstrated that blood-sucking insects protect their physiological integrity either by synthesising heat-shock proteins or by means of thermoregulatory mechanisms. In this work, we describe the first thermoregulatory mechanism in a tick species, Ornithodoros rostratus. By performing real-time infrared thermography during feeding on mice we found that this acarian eliminates big amounts of fluid (urine) through their coxal glands; this fluid quickly spreads over the cuticular surface and its evaporation cools-down the body of the tick. The spread of the fluid is possible thanks to capillary diffusion through the sculptured exoskeleton of Ornithodoros. We discuss our findings in the frame of the adaptive strategies to cope with the thermal stress experienced by blood-sucking arthropods at each feeding event on warm-blooded hosts.


Clonal diversity of extended-spectrum beta-lactamase producing Escherichia coli isolates in fecal samples of wild animals.
The clonal diversity of extended-spectrum-β-lactamase (ESBL)-producing Escherichia coli isolates from nine different species of wild animals from distinct regions of Portugal and Spain and their content in replicon plasmids were analyzed. Among the initial 53 ESBL-producing E. coli isolates that were studied (from previous studies), 28 were selected, corresponding to different animal origins with distinct ESBL types and pulsed-field gel electrophoresis (PFGE) patterns. These 28 isolates produced different ESBLs ascribed to the following families: CTX-M, SHV and TEM. The isolates were classified into three phylogenetic groups: B1 (n = 11), A (n = 10) and D (n = 7). The seven E. coli of phylogroup D were then typed by multilocus sequence typing and ascribed to four distinct sequence types: ST117, ST115, ST2001 and ST69. The clonal diversity and relationship between isolates was studied by PFGE. Lastly, the plasmids were analyzed according to their incompatibility group using the PCR-based-replicon-typing scheme. A great diversity of replicon types was identified, with up to five per isolate. Most of the CTX-M-1 and SHV-12 producing E. coli isolates carried IncI1 or IncN replicons. The diversity of ESBL-producing E. coli isolates in wild animals, which can be disseminated in the environment, emphasizes the environmental and health problems that we face nowadays.


Recent advances in the induced pluripotent stem cell-based skin regeneration.
Skin regeneration has been a challenging clinical problem especially in cases of chronic wounds such as diabetic foot ulcers, and epidermolysis bullosa-related skin blisters. Prolonged non-healing wounds often lead to bacterial infections increasing the severity of wounds. Current treatment strategies for chronic wounds include debridement of wounds along with antibiotics, growth factors, and stem cell transplantation therapies. However, the compromised nature of autologous stem cells in patients with comorbidities such as diabetes limits the efficacy of the therapy. The discovery of induced pluripotent stem cell (iPSC) technology has immensely influenced the field of regenerative therapy. Enormous efforts have been made to develop integration-free iPSCs suitable for clinical therapies. This review focuses on recent advances in the methods and reprogramming factors for generating iPSCs along with the existing challenges such as genetic alterations, tumorigenicity, immune rejection, and regulatory hurdles for the clinical application of iPSCs. Furthermore, this review also highlights the benefits of using iPSCs for the generation of skin cells and skin disease modeling over the existing clinical therapies for skin regeneration in chronic wounds and skin diseases.


Ultrasonic diagnostics: a look into the future.
The widespread introduction of ultrasonic diagnostics will depend upon the availability of simple-to-use, cost effective instruments producing results which are easily interpreted. There is scope for improvement in conventional two-dimensional scanning systems. Real-time display would allow the operator more effectively to interact with the machine. The identification of tissues may become possible. Blood flow volumes may be measured by the Doppler method, and hybrid Doppler/pulse-echo systems may have applications in the study of cardiovascular disease. Monitoring of foetal breathing may become routine. Time delay spectrometry holds some promise, but holography does not. Piezo-electric plastic transducers may be introduced.


Extracorporeal Shockwave Therapy Plus Rehabilitation for Patients With Chronic Plantar Fasciitis Might Reduce Pain and Improve Function but Still Not Lead to Increased Activity: A Case-Series Study With Multiple Outcome Measures.
Plantar fasciitis is a common cause of plantar-aspect heel pain. Although many patients will improve, a proportion will have ongoing and sometimes debilitating symptoms. Evidence from randomized controlled trials has shown that extracorporeal shockwave therapy (ESWT) results in benefits in treating pain. However, uncertainties remain whether these benefits translate to improvements in overall function. The present prospective case series examined the results from 35 patients with chronic plantar fasciitis who had undergone a course of ESWT in addition to a graded rehabilitation program. Of the 35 subjects, 34% were male, and the median age was 50.9 years. The duration of symptoms before ESWT was 24 months. The results of the present case series demonstrated statistically significant improvements in measures of self-reported "average pain" from a median of 7.0 of 10 at baseline to 5.0 of 10 at 3 months (p < .001) and of "worst pain" from 9.0 of 10 at baseline to 7.0 of 10 at 3 months (p < .001). In addition, significant improvements were found in several validated patient-rated outcome measures of local foot/ankle function but not in overall markers of health, anxiety/depression scores, or activity levels, despite the improvements in pain. No statistically significant correlations were found between gender, age, or chronicity of symptoms and the improvements seen. No significant side effects occurred in the present study. The results of our series support the use of ESWT for patients with chronic plantar fasciitis for local pain symptoms; however, uncertainties remain regarding global benefits to health.


Problem-specific coding systems.
The data found in medical records, particularly in problem-oriented records, could yield very useful information for many aspects of the health care delivery system. Unless the tools and methods used in the analysis of health care data are very carefully designed, the potential value of this information cannot be fully realized. Coding is a critical step in the process of converting medical record data into useful information. This paper presents six criteria as design goals for a standard, comprehensive, coherent set of nomenclatures, classifications, taxonomies, and corresponding coding schemes for the analysis of medical and administrative record data. These goals are: compatibility with established routine operating procedure, health care problem specificity, hierarchical structure, economy through articulation of the components with one another and through the similarities in various aspects of health care delivery, and adaptability to change.