Epidemiological and clinical complexity of amoxicillin-clavulanate-resistant Escherichia coli.
Two hundred twelve patients with colonization/infection due to amoxicillin-clavulanate (AMC)-resistant Escherichia coli were studied. OXA-1- and inhibitor-resistant TEM (IRT)-producing strains were associated with urinary tract infections, while OXA-1 producers and chromosomal AmpC hyperproducers were associated with bacteremic infections. AMC resistance in E. coli is a complex phenomenon with heterogeneous clinical implications.


Insight into antimicrobial susceptibility and population structure of contemporary human Enterococcus faecalis isolates from Europe.
To investigate antimicrobial susceptibility and clonal relatedness of Enterococcus faecalis human isolates recovered recently (2006-09) in six European countries. Antimicrobial susceptibility of 386 isolates from Denmark, Germany, Norway, Poland, Spain and The Netherlands, from hospital infections (223 isolates), carriage (82 isolates) and from colonization in the community (81 isolates) was determined by the broth microdilution method. Clonal relatedness of isolates was assessed by multilocus sequence typing. All isolates were susceptible to benzylpenicillin, ampicillin, linezolid, tigecycline and daptomycin. Non-susceptibility to tetracycline (77.6%), rifampicin (57.3%), ciprofloxacin (51.2%), aminoglycosides (43.3% high-level gentamicin resistance, 40.0% high-level streptomycin resistance) was frequent among hospital isolates, while non-susceptibility to glycopeptides was rare and associated mostly with vanA. Multidrug resistance was found in 59.7% of hospital isolates and 16.1% of community isolates. Isolates were classified into 105 sequence types (STs), of which 21 STs, representing more than half of the collected isolates (53.9%), grouped with 6 large E. faecalis clonal complexes (CCs; CC2, CC16, CC21, CC30, CC40 and CC87). Two of these, CC2 (frequently recovered in Spain and The Netherlands) and CC87 (prevalent in Poland), were found almost exclusively in hospitals and included the highest proportion of multiresistant isolates. While hospital-acquired E. faecalis in Europe remains susceptible to ampicillin and glycopeptides, the high prevalence of strains that are highly resistant to aminoglycosides excludes these antibiotics from combination therapies. Genotyping revealed that nosocomial infections by multiresistant E. faecalis are largely caused by only a few hospital-associated clones.


Amebic liver abscess: MR imaging.
Hepatic magnetic resonance (MR) imaging was performed in 12 patients with 13 amebic liver abscesses. While no specific image or intensity pattern was noted, most lesions were round or oval with smooth, well-defined margins; had decreased signal intensity compared with that of liver parenchyma on T1-weighted images and increased signal intensity on T2-weighted images; and had prominent, often multiple rims of variable signal intensity. Signal homogeneity within the abscess was present more often on T1- than on T2-weighted images. Diaphragmatic disruption was seen in two cases on coronal MR images. An amebic empyema was differentiable from sympathetic pleural effusions by its hyperintensity on both T1- and T2-weighted images. In patients who also underwent computed tomography (CT) or ultrasonography (US), no lesion was missed with any modality, and except for shape, no consistent features were found among images obtained with the different modalities. The data suggest that CT, US, and MR imaging are comparably effective in the detection of amebic abscess.


Hazard Analyses of Foods Prepared by Inhabitants Along the Peruvian Amazon River.
Hazard analyses of food preparation practices were conducted in two households in Indiana (a settlement along the Peruvian Amazon River), in a household in a cluster of about a half dozen houses up river, and in three households in Belen (a district near Iquitos), Peru. These analyses consisted of watching all steps of the operation, recording temperatures throughout all these steps, and collecting samples of food and testing them for common foodborne pathogens and indicator organisms. Foods prepared included rice, plantains, yuca, dry fish, fresh fish, beef, and chicken. During cooking, foods attained temperatures of at least 93.3°C; they usually boiled. Such time-temperature exposure would kill vegetative forms of foodborne pathogenic bacteria, but not heat-resistant spores. When cooked foods were leftover, they were kept either on tables or on the unheated stoves or grills on which they were cooked. During this interval, at the prevailing ambient temperature and high humidity of the jungle region, conditions were such that considerable microbial growth could occur. Time of exposure, however, limited counts to the 105-106 level. In the evening, foods were only mildly reheated, if reheated at all, so temperatures were not attained in the center regions of the food that would have killed microorganisms that had multiplied during the holding period. Hence, the primary critical control point is holding between cooking and serving, but cooking and reheating are critical control points also.


BAR502, a dual FXR and GPBAR1 agonist, promotes browning of white adipose tissue and reverses liver steatosis and fibrosis.
Non-alcoholic steatohepatitis (NASH) is a highly prevalent chronic liver disease. Here, we have investigated whether BAR502, a non-bile acid, steroidal dual ligand for FXR and GPBAR1, reverses steato-hepatitis in mice fed a high fat diet (HFD) and fructose. After 9 week, mice on HFD gained ≈30% of b.w (P < 0.01 versus naïve) and were insulin resistant. These overweighting and insulin resistant mice were randomized to receive HFD or HFD in combination with BAR502. After 18 weeks, HFD mice developed NASH like features with severe steato-hepatitis and fibrosis, increased hepatic content of triacylglycerol and cholesterol and expression of SREPB1c, FAS, ApoC2, PPARα and γ, α-SMA, α1 collagen and MCP1 mRNAs. Treatment with BAR502 caused a ≈10% reduction of b.w., increased insulin sensitivity and circulating levels of HDL, while reduced steatosis, inflammatory and fibrosis scores and liver expression of SREPB1c, FAS, PPARγ, CD36 and CYP7A1 mRNA. BAR502 increased the expression of SHP and ABCG5 in the liver and SHP, FGF15 and GLP1 in intestine. BAR502 promoted the browning of epWAT and reduced liver fibrosis induced by CCl4. In summary, BAR502, a dual FXR and GPBAR1 agonist, protects against liver damage caused by HFD by promoting the browning of adipose tissue.


Dental plaque removal by cavitational activity during ultrasonic scaling.
Photomicrographs of the surfaces of recently extracted teeth stained with erythrosin dye were obtained. The stained surfaces were treated with an ultrasonic scaler using two different types of scaling tips driven by the same instrument operated at a medium power setting. Small areas of stained plaque removal occurred when the stationary scaling tip was operated without water cooling. Additional areas of removal were observed where a water coolant was present, which were larger than those produced by the non-water cooled tip. These additional areas were influenced by the type of scaling tip used, it's orientation to the tooth surface and it's displacement amplitude. Cavitational activity in the cooling water supply of the ultrasonic scaler is able to remove dental plaque from tooth surfaces and may be a useful adjunct to the mechanical action of the instrument.


Self-assembly of symmetric finite-size DNA nanoarrays.
We report a novel and cost-effective strategy to self-assemble finite-size DNA nanoarrays. This strategy takes advantage of the geometric symmetry of the DNA nanostructures. In general, to construct a 2D array with a total of N tiles containing Cm symmetry, where m = 2, 3, 4, or 6, the number of unique tiles the fixed-size array requires is N/m, if N/m is an integral number, or Int(N/m) + 1, if N/m is an nonintegral number. We herein demonstrate two examples of fixed-size arrays with C2 and C4-fold symmetry.dagger


Polyethylene Liner Dissociation after Reverse Shoulder Arthroplasty Dislocation: a Case Series.
Reverse shoulder arthroplasty is a common procedure. While dislocation is a common complication, there are few reports documenting dislocation with associated polyethylene liner dissociation from the humeral tray. There have been 4 instances of this occurrence over 9 years at our institution, and there are specific radiographic clues for diagnosis. Recognizing polyethylene liner dissociation on imaging is important prior to treatment. In a "routine" dislocation, closed reduction may be attempted but when the polyethylene is dissociated, open reduction is the only treatment option, as closed reduction can damage the components. Dislocations with polyethylene dissociation may not be initially recognized, prompting a non-operative period leading to wear and metallosis. These 4 cases demonstrate key findings present on imaging to recognize the difference between a dislocation with and without polyethylene liner dissociation, namely the subluxation appearance rather than dislocation.


Risk Factors for Reoperation After Arterial Switch Operation.
The timing and nature of and risk factors for reoperation after the arterial switch operation in the setting of d-transposition of the great arteries requires further elucidation. A total of 403 patients who underwent arterial switch operation from 1986 to 2017 were reviewed. Institutional preference was for pulmonary artery reconstruction using a pantaloon patch of fresh autologous pericardium. The targets for coronary artery reimplantation were identified by intermittent root distension. Multivariable analysis was used to identify risk factors for reoperation. Median follow-up was 8.6 years (interquartile range [IQR]: 2-16.9). Pulmonary arterioplasty was the most common reoperation (n = 11, 2.7%) at 3.3 years (IQR: 1.4-11.4) postoperatively. Subvalvar right ventricular outflow tract reconstruction (RVOTR) was required in nine (2.2%) patients at 2.5 years (IQR: 1.1-5.3) postoperatively. Aortic valve repair or replacement (AVR/r) was required in seven (1.7%) patients at 13.6 years (IQR: 10.0-15.8) postoperatively. Aortic root replacement (ARR) and Coronary Artery Bypass Graft/coronary patch arterioplasty were required in five (1.2%) patients each at 13.6 years (IQR: 11.0-15.3) and 11.3 years (IQR: 2.3-13.6) postoperatively, respectively. Taussig-Bing anomaly was a risk factor for any reoperation (P = .034). Risk factors for specific reoperations included ventricular septal defect for AVR/r (P = .038), Taussig-Bing anomaly for RVOTR (P = .004), and pulmonary artery banding for ARR (P = .028). Pantaloon patch pulmonary artery reconstruction and intermittent neo-aortic root distension during coronary reimplantation have minimized respective outflow tract reoperations. Certain anatomic subsets carry different risks for late reoperation, and pulmonary artery and/or RVOT reinterventions tend to occur sooner than aortic reinterventions. Special attention to these higher risk subpopulations will be critical to optimizing lifelong outcomes.


Contrast perception in digitized panoramic radiographs compared with their film-based origin.
We sought to compare the visual image quality of film-based and digitized panoramic radiographs through use of a hole-containing test wedge. An aluminum wedge containing 100 cells, of which 90 were given shallow holes, was exposed in the film-based Orthophos CD panoramic unit. Two radiographs subjectively exhibiting optimum contrast were selected. Films were digitized with a charge-coupled device flatbed scanner at 300 dpi. Films and digitized images were rated cellwise by 2 similar groups of 50 observers each with respect to spot perception. The mean sensitivity was 0.26 +/- 0.09 for film and 0.20 +/- 0.07 for digitized images (P =.000), with a pronounced decline in the latter in regions of high background density. The average specificity was 0.93 +/- 0.07 for film versus 0.92 +/- 0.08 for digitized images (P = 0.213). Film yielded a significantly higher sensitivity, but this absolute difference was actually small compared with that of the digitized images.


Studies on synthetic approaches to 1H- and 2H-indazolyl derivatives.
Synthetic approaches designed to provide 1H- and 2H-indazolyl derivatives of potential biological interest are reported. Special emphasis has been placed on the characterization of indazolylpyridinium products generated from reactions between indazole and 4-chloro-1-methylpyridinium iodide under various conditions. A stable mixture consisting of 3 parts of the 1H-isomer 9 to 1 part of the 2H-isomer 10 was obtained at room temperature in the presence of the base 2,2,6,6-tetramethylpiperidine (TMP). The same reaction at 60 degrees C gave only the 1H-isomer 9. At 100 degrees C in the absence of TMP only the 2H-isomer 10 was formed. The isomerization of 10 to 9 was found to proceed quantitatively at 60 degrees C but only in the presence of TMP. The effects of temperature and base on the course of these reactions are rationalized in terms of kinetic and thermodynamic parameters.


mHealth Interventions for Contraceptive Behavior Change in the United States: A Systematic Review.
Ensuring people have access to their preferred method of contraception can be key for meeting their reproductive goals. A growing number of mHealth interventions show promise for improving access to contraception, but no literature review has identified the effects of mHealth interventions among both adolescents and adults in the United States. The purpose of this systematic review was to describe the format, theoretical basis, and impact of mHealth interventions for contraceptive behavior change (contraceptive initiation and continuation) among people of all ages in the US. A systematic review of the literature was conducted using six electronic databases guided by Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) guidelines. Data on study design, frequency, duration, mHealth modality, contraceptive method, behavior change theory, and behavioral outcome were extracted to facilitate comparison. Eighteen studies met eligibility criteria. The majority (11; 61%) used SMS (short message service). Twelve studies focused on contraceptive initiation, most (n = 8) of which also measured continued use over time. The remaining six interventions focused on continuation alone, generally through appointment reminders. Very little contraceptive behavior change was identified across studies. Current mHealth interventions may hold promise for some health areas but there is little evidence that they change contraceptive behavior. Future mHealth interventions should focus on assessing person-centered outcomes, including satisfaction, side effects, and reasons for discontinuation, to best support people to use their preferred contraceptive method.


Lower risk of postinfarct rupture in mouse heart overexpressing beta 2-adrenergic receptors: importance of collagen content.
This paper addresses whether the enhanced left ventricular (LV) contractility and heart rate, seen in transgenic mice overexpressing beta -adrenergic receptor in the heart, might raise the incidence of LV rupture after myocardial infarct. Transgenic and wild-type mice underwent left coronary artery occlusion. Postinfarct deaths that occurred 1-7 days after surgery were analyzed. Hemodynamics, morphologic parameters, and collagen content in the LV were determined. A significantly lower incidence of LV rupture was observed in transgenic than in wild-type mice 3-5 days after myocardial infarct (2.5 versus 19.7%, p < 0.05), despite a similar infarct size between the two groups and better hemodynamic function in transgenic mouse hearts. Morphologic analysis showed a more severe infarct expansion in wild-type versus transgenic mice or in mice dying of rupture versus those that died of acute heart failure. Collagen content was higher in the LV of sham-operated transgenic than wild-type mice (p < 0.01) with both type I and type III collagen elevated. Such difference in collagen content between transgenic and wild-type mice was maintained in noninfarcted and infarcted LV. In conclusion, transgenic mice overexpressing beta -adrenergic receptor had a lower risk of cardiac rupture during the acute phase after infarction despite the markedly enhanced LV contractility and heart rate. As a hyperdynamic function due to beta-adrenergic activation would likely increase the risk of cardiac rupture and infarct expansion, the lack of rupture in this transgenic mouse model suggests that the interstitial collagen level is a more important factor than functional status in the pathogenesis of rupture and infarct expansion.


Surface Passivation of Layered MoSe2 via van der Waals Stacking of Amorphous Hydrocarbon.
Development of efficient surface passivation methods for semiconductor devices is crucial to counter the degradation in their electrical performance owing to scattering or trapping of carriers in the channels induced by molecular adsorption from the ambient environment. However, conventional dielectric deposition involves the formation of additional interfacial defects associated with broken covalent bonds, resulting in accidental electrostatic doping or enhanced hysteretic behavior. In this study, centimeter-scaled van der Waals passivation of transition metal dichalcogenides (TMDCs) is demonstrated by stacking hydrocarbon (HC) dielectrics onto MoSe2 field-effect transistors (FETs), thereby enhancing the electric performance and stability of the device, accompanied with the suppression of chemical disorder at the HC/TMDCs interface. The stacking of HC onto MoSe2 FETs enhances the carrier mobility of MoSe2 FET by over 50% at the n-branch, and a significant decrease in hysteresis, owing to the screening of molecular adsorption. The electron mobility and hysteresis of the HC/MoSe2 FETs are verified to be nearly intact compared to those of the fabricated HC/MoSe2 FETs after exposure to ambient environment for 3 months. Consequently, the proposed design can act as a model for developing advanced nanoelectronics applications based on layered materials for mass production.


Prevention of 20-methylcholanthrene-induced sarcoma by a mistletoe extract, Iscador.
Iscador, an extract from the semi-parasitic plant Viscum album, was found to inhibit 20-methylcholanthrene-induced carcinogenesis in mice. Intraperitoneal administration of Iscador (1 mg/dose) twice weekly for 15 weeks could completely inhibit 20-methylcholanthrene-induced sarcoma in mice and protect these animals from tumour-induced death. Iscador was found to be effective even at lowered doses. After administration of 0.166, 0.0166 and 0.00166 mg/dose 67, 50 and 17% of animals respectively did not develop sarcoma.


Human-wildlife conflict in the surrounding districts of Alage College, Central Rift Valley of Ethiopia.
The study was conducted between September 2018 and March 2019 to investigate the nature and extent of human-wildlife conflict (HWC) in the surrounding area of Alage College, the Central Rift Valley of Ethiopia, and to assess the perception of the local people to wildlife. For data collection, a total of 140 household (HH) heads were selected randomly for interviews from nine villages using structured and semi-structured questionnaires. Moreover, focus group discussion, key informant interviews, and personal observation were carried out to obtain additional information. Descriptive statistics, Chi-square test (2 tailed), one-way analysis of variance, Pearson correlation coefficient, and Likert scale statements were used to analyze the data. Based on, 66 (47.1%) of the respondents, HH heads, the local people of the area experienced livestock predation leading to HWC. Whereas, (40.7%, n = 57) of the respondents perceived both crop damage and livestock predation as a cause of conflict. A total of 932.43 total livestock unit of livestock and 218 dogs' losses were reported by HH due to predators over the last 5 years. Thus, the largest number of livestock (89.9%) and dogs (100%) attacks was happened due to spotted hyenas. Nearly half of the respondents (49.3%, n = 69) ranked warthogs as the primary crop raiders, while the majority of respondents (82.1%, n = 115) reported maize as a severely damaged cereal crop. More than half, (57.1%, n = 80) of respondents used different methods simultaneously to minimize damage caused by wild animals. About half, (48.6%, n = 68) of respondents had a negative attitude toward wildlife conservation. The level of education and amount of money imposed as a penalty for illegal grazing were affecting the local community's attitudes to wildlife conservation. Using effective methods to reduce damage and loss to crops, including improved livestock husbandry and creating better awareness to the local community could make the locals actor of conservation.


A comparison of long-term ventilatory support alternatives from the perspective of the patient and care giver.
Tracheostomy, noninvasive intermittent positive pressure ventilation (IPPV) methods, and body ventilators can be used for ventilatory support. These methods vary with respect to safety, convenience, effect on speech, sleep, swallowing, appearance, comfort, and general patient acceptability. The purpose of this study was to determine patient and care giver ventilatory support preferences concerning these quality of life issues. One hundred sixty-eight predominantly neuromuscular patients with greater than 1 month of experience in the use of both tracheostomy and noninvasive methods were surveyed. At the time of the study they had a mean age of 54.7 +/- 11.4 years, were using ventilatory aids a mean of 17.1 +/- 6.5 h/d, and had been doing so for 22.7 +/- 13.1 consecutive years. The 59 respondents who switched from tracheostomy to noninvasive aids significantly preferred the latter for every item and unanimously preferred them over all. The 76 respondents switched from body ventilator use to tracheostomy IPPV significantly preferred use of the former for appearance and speech and the latter for sleep and security and had no other significant preferences. However, the 35 respondents switched from a regimen of noninvasive aids including noninvasive IPPV to tracheostomy IPPV significantly preferred the noninvasive aids for each item except for sleep, swallowing, and security for which there were no significant preferences. We conclude that whether patients switched to or from noninvasive ventilatory support regimens which include noninvasive IPPV, noninvasive methods are considered by patients to be more convenient, to have less untoward effects on speech, appearance and comfort, and are preferred overall to tracheostomy IPPV.


[Positron emission tomography (PET) for diagnosis and monitoring of treatment for urological tumors].
Positron emission tomography (PET) using ((18)F)2-fluoro-D-2-desoxyglucose (FDG) has been shown to be a highly sensitive and specific imaging modality in the diagnosis of primary and recurrent tumors and in the control of therapies in numerous non-urologic cancers. It was the aim of this review to validate the significance of PET as a diagnostic tool in malignant tumors of the urogenital tract. A systematic review of the current literature concerning the role of PET for malignant tumors of the kidney, testicles, prostate, and bladder was carried out. The role of FDG PET for renal cell cancer can be seen in the detection of recurrences after definitive local therapy and metastases. The higher sensitivity of PET in comparison to other therapeutic modalities (CT, ultrasound, MRI) in recurrent and metastatic renal cell cancer suggests a supplemental role of this diagnostic procedure to complement other imaging modalities.The clinical value of PET is established for the identification of vital tumor tissue after chemotherapy of seminomatous germ cell tumors. This diagnostic method has little significance for primary tumor staging and diagnosis of non-seminomatous germ cell tumor because of the high probability of false-negative results in adult teratomas. FDG PET is not sensitive enough in the diagnosis of primary or recurrent tumors in prostate or bladder cancer. Also PET did not prove to be superior to conventional bone scintigram in the detection of mostly osteoblastic metastases in prostate cancer. The recent use of alternative tracers, which are partly not eliminated by urinary secretion (acetate, choline) has increased the sensitivity and specificity of PET also in this tumor entity so that further clinical investigations are needed to validate these technical modifications in their significance for this imaging modality. PET appears to be sufficiently evaluated only for the diagnostic follow-up of patients with seminomatous germ cell tumors after chemotherapy to regard it is the diagnostic tool of first choice. For all other tumors of the urogenital tract this proof is still awaited.


Forging the Basis for Developing Protein-Ligand Interaction Scoring Functions.
In structure-based drug design, scoring functions are widely used for fast evaluation of protein-ligand interactions. They are often applied in combination with molecular docking and de novo design methods. Since the early 1990s, a whole spectrum of protein-ligand interaction scoring functions have been developed. Regardless of their technical difference, scoring functions all need data sets combining protein-ligand complex structures and binding affinity data for parametrization and validation. However, data sets of this kind used to be rather limited in terms of size and quality. On the other hand, standard metrics for evaluating scoring function used to be ambiguous. Scoring functions are often tested in molecular docking or even virtual screening trials, which do not directly reflect the genuine quality of scoring functions. Collectively, these underlying obstacles have impeded the invention of more advanced scoring functions. In this Account, we describe our long-lasting efforts to overcome these obstacles, which involve two related projects. On the first project, we have created the PDBbind database. It is the first database that systematically annotates the protein-ligand complexes in the Protein Data Bank (PDB) with experimental binding data. This database has been updated annually since its first public release in 2004. The latest release (version 2016) provides binding data for 16 179 biomolecular complexes in PDB. Data sets provided by PDBbind have been applied to many computational and statistical studies on protein-ligand interaction and various subjects. In particular, it has become a major data resource for scoring function development. On the second project, we have established the Comparative Assessment of Scoring Functions (CASF) benchmark for scoring function evaluation. Our key idea is to decouple the "scoring" process from the "sampling" process, so scoring functions can be tested in a relatively pure context to reflect their quality. In our latest work on this track, i.e. CASF-2013, the performance of a scoring function was quantified in four aspects, including "scoring power", "ranking power", "docking power", and "screening power". All four performance tests were conducted on a test set containing 195 high-quality protein-ligand complexes selected from PDBbind. A panel of 20 standard scoring functions were tested as demonstration. Importantly, CASF is designed to be an open-access benchmark, with which scoring functions developed by different researchers can be compared on the same grounds. Indeed, it has become a popular choice for scoring function validation in recent years. Despite the considerable progress that has been made so far, the performance of today's scoring functions still does not meet people's expectations in many aspects. There is a constant demand for more advanced scoring functions. Our efforts have helped to overcome some obstacles underlying scoring function development so that the researchers in this field can move forward faster. We will continue to improve the PDBbind database and the CASF benchmark in the future to keep them as useful community resources.


In Re Doe, 12 November 1987.
The natural mother of profoundly retarded woman in custody of the state challenged an order of the Family Court that the pregnancy of her daughter be terminated. The order was made pursuant to the petition of the executive director of the Division of Retardation of the Rhode Island Department of Mental Health, Retardation, and Hospitals. The Court rejected the challenge, holding that the conclusion of the Family Court that the young woman, if she had been able, would have decided to have an abortion, was correct, particularly in light of the fact that she was suffering from seizure disorder and cerebral palsy.


Accumulation of anthracenyl-amino acid topoisomerase I and II inhibitors in drug-sensitive and drug-resistant human ovarian cancer cell lines determined by high-performance liquid chromatography.
Anthracenyl amino acid/dipeptide conjugates (AADC) represent novel structures rationally designed for their DNA-binding properties. A high-performance liquid chromatography method is described for simultaneous determination of five compounds that exhibit novel mechanisms of action as topoisomerase I and II inhibitors. The method uses an Apex ODS-2 column and a mobile phase of 0.25 M ammonium acetate/trifluoroacetic acid (pH 3) in methanol with gradient elution. Selective detection is achieved by monitoring at 545 nm, with limits of detection ranging between 2 and 4 ng on the column. AADC are recovered from cell sonicates by solid-phase extraction using C2 cartridges, with extraction efficiencies ranging from 84% to 95%. Drug uptake studies were performed with three active compounds in the human ovarian cancer cell line A2780 and its multi-drug-resistant counterpart 2780AD. Marked differences were observed in the pattern of cellular accumulation produced by each compound. NU/ICRF 505 (tyrosine derivative) was taken up most avidly, reaching plateau levels of 4000 pmol/10(6) cells after 2 h, with no difference being apparent between A2780 and 2780AD. NU/ICRF 510 (arginine derivative) accumulated slowly in A2780, failing to achieve an equilibrium after 4 h, and appeared to be completely excluded from 2780AD. NU/ICRF 500 (serine derivative) was most rapidly taken up by A2780, producing a plateau of 800 pmol/10(6) cells after only 30 min with approximately 3-fold less accumulation in 2780AD. These results are correlated to the chemosensitivity of the two cell lines to the three compounds.


Epigenetic dynamics: role of epimarks and underlying machinery in plants exposed to abiotic stress.
Abiotic stress induces several changes in plants at physiological and molecular level. Plants have evolved regulatory mechanisms guided towards establishment of stress tolerance in which epigenetic modifications play a pivotal role. We provide examples of gene expression changes that are brought about by conversion of active chromatin to silent heterochromatin and vice versa. Methylation of CG sites and specific modification of histone tail determine whether a particular locus is transcriptionally active or silent. We present a lucid review of epigenetic machinery and epigenetic alterations involving DNA methylation, histone tail modifications, chromatin remodeling, and RNA directed epigenetic changes.


[Neoadjuvant chemotherapy and surgical treatment in advanced stages of non-small cell lung cancer].
Lung cancer takes first place in both incidence and mortality in the Czech Republic. This is associated with the disease being diagnosed in late stages, which limits the possibility of radical therapy. Five-year survival of patients operated on with stage IIIA is low and doesn't even reach 20%. These poor results and the development of systemic chemotherapy in the 1990's led to an effort to treat locally advanced disease by administering chemotherapy before the surgical procedure- induction chemotherapy. Its benefit, however, unlike that of adjuvant chemotherapy, remains unclear. To analyze and compare the results between a set of patients with non-small cell lung cancer (NSCLC) with stage III A-B, operated on at the I. Department of Surgery at the University Hospital and Palacky Medical Faculty in Olomouc between the years 2000-2008, who underwent preoperative chemotherapy with the results of patients with stage III A-B diagnosed after the operation based on histological findings. Three- and five-year survivals, as well as survival median, were evaluated in both groups. A statistically significant difference in survival between the two groups was not observed. Neoadjuvant chemotherapy remains controversial in the treatment of NSCLC. The initially promising results have not been unequivocally confirmed in later studies and its role remains a question to be answered in future extensive randomized studies.


Dissecting aneurysm at the A1 segment of the anterior cerebral artery presenting as visual loss and visual field defect.
Intracranial dissecting aneurysms mainly occur in the territory of the vertebrobasilar system. Dissecting aneurysms confined to the anterior cerebral artery are rare, and the presentations are usually of either subarachnoid hemorrhage or cerebral infarction. Here, we report a unique case of a dissecting aneurysm of the anterior cerebral artery presenting as a visual field defect. After surgical decompression, visual symptoms recovered.


Small intestinal sampling capsule for inflammatory bowel disease type detection and management.
Although serum and fecal biomarkers (e.g., lactoferrin, and calprotectin) have been used in management and distinction between inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS), none are proven to be a differential diagnostic tool between Crohn's disease (CD) and ulcerative colitis (UC). The main challenge with laboratory-based biomarkers in the stool test is the inability to indicate the location of the disease/inflammation in the gastrointestinal (GI) tract due to the homogenous nature of the collected fecal sample. For the first time, we have designed and developed a battery-free smart capsule that will allow targeted sampling of inflammatory biomarkers inside the gut lumen of the small intestine. The capsule is designed to provide a simple and non-invasive complementary tool to fecal biomarker analysis to differentiate the type of IBD by pinpointing the site of inflammatory biomarkers secretion (e.g., small or large bowel) throughout the GI tract. The capsule takes advantage of the rapid change from an acidic environment in the stomach to higher pH levels in the small intestine to dissolve a pH-sensitive polymeric coating as a means to activate the sampling process of the capsule within the small intestine. A swelling polyacrylamide hydrogel is placed inside the capsule as a milieu to collect the sampled GI fluid while also providing the required mechanical actuation to close the capsule once the sampling is completed. The hydrogel component along with the collected GI fluid can be easily obtained from the capsule through the screw-cap design for further extraction and analysis. As a proof of concept, the capsule's performance in sampling and extraction of bovine serum albumin (BSA) and calprotectin - a key biomarker of inflammation - was assessed within the physiologically relevant ranges. The ratio of extracted biomarkers relative to that in the initial sampling environment remained constant (∼3%) and independent of the sampling matrix in both in vitro and ex vivo studies. It is believed that the demonstrated technology will provide immediate impact in more effective IBD type differential diagnostic and treatment strategies by providing a non-invasive assessment of inflammation biomarkers profile throughout the digestive tract.


[The investigation of humic acid by surface-enhanced Raman spectroscopy].
Humic acid (HA), which are organic compounds widely existing in the oceans, rivers and soil, has important significance for the environmental monitoring of soil and water. In this paper, ai ming at the problem of Surface-enhanced Raman Spectroscopy (SERS) applying for HA detection in water, the characteristics of the humic acid on silver colloids was studied by means of SERS. The influence of laser irradiation time, HA concentrations and pH value on the surface-enhanced effects of HA were investigated. The experimental results show that the SERS spectra of HA is ideal when the laser irradiation time between 20-30 min. The SERS of different HA concentrations was detected. It was found that the relative intensity at 1 379 cm(-1) increased as a linear function of the concentration of HA with correlation coefficient R2 of 0.993. The SERS of HA was found to be very sensitive to pH, the SERS spectra of HA was very weak at neutral pH, but at acidic pH and alkaline pH a remarkable increase of SERS intensity occurred. The SERS of HA in running water was detected too. The experimental results show that it is feasible to detect HA in natural water by means of surface-enhanced Raman spectroscopy.


Using Greedy algorithm: DBSCAN revisited II.
The density-based clustering algorithm presented is different from the classical Density-Based Spatial Clustering of Applications with Noise (DBSCAN) (Ester et al., 1996), and has the following advantages: first, Greedy algorithm substitutes for R(*)-tree (Bechmann et al., 1990) in DBSCAN to index the clustering space so that the clustering time cost is decreased to great extent and I/O memory load is reduced as well; second, the merging condition to approach to arbitrary-shaped clusters is designed carefully so that a single threshold can distinguish correctly all clusters in a large spatial dataset though some density-skewed clusters live in it. Finally, authors investigate a robotic navigation and test two artificial datasets by the proposed algorithm to verify its effectiveness and efficiency.


Acute remodeling of an adjoining aneurysm after endovascular treatment of a ruptured splanchnic arterial aneurysm: a case of clinically diagnosed segmental arterial mediolysis.
A 61-year-old woman with multiple splanchnic arterial aneurysms (SAAs) was transferred to our hospital in a state of shock. She underwent coil embolization under the diagnosis of ruptured pancreaticoduodenal artery aneurysm. Follow-up computed tomography performed 2 weeks later showed rapid enlargement of a gastric artery aneurysm, and she underwent an additional embolization. Atherosclerotic, inflammatory or hereditary causes were excluded, and the patient was clinically diagnosed with segmental arterial mediolysis accompanied by multiple SAAs, one of which showed acute remodeling after endovascular treatment.


Resistance and the control of sheep ectoparasites.
For about 100 years Australian woolgrowers have used a variety of chemicals to control blowflies, lice and other ectoparasites of sheep. While the chemicals have changed considerably the application technology has not changed very much at all. Chemicals registered for use on sheep have paralleled the evolution of synthetic insecticides with the unfortunate consequence of the development of resistance in the Australian sheep blowfly, Lucilia cuprina, following closely behind. Organochlorine (dieldrin) resistance peaked at about 70% in 1958 when unacceptable residues in meat and wool forced their withdrawal. Organophosphate (OP) resistance appeared in 1965. With no alternative insecticide classes until 1979, OP resistance reached near fixation levels by the early 1970s and has remained unchanged. OP resistance has reduced the protection period from over 16 weeks to about 6 weeks. Moreover, resistance has decreased the effectiveness of many flystrike dressings to unacceptably low levels. OPs are still very effective against sheep body lice, Bovicola ovis but control is hampered by inadequate application via plunge or shower dipping. Synthetic pyrethroid (SP) pour-on products were released in 1981 but resistance developed by 1985 and many woolgrowers were unable to eradicate lice with pour-on products. Highest Resistance Factors at this time were only about 26 x but this was sufficient to prevent pour-ons working efficiently. By 1991 a population from Hartley in NSW was found to be 642 x resistant to cypermethrin with side-resistance conferred to the other SPs. SP resistance was partially suppresible by piperonyl butoxide but field trials suggested that the resulting improvement in efficacy was not sufficient to be commercially attractive. OPs remain very effective if applied correctly and the release of ivermectin and 2 benzoylphenyl urea products significantly improves the prospects for resistance management. However the increasing environmental concern about the persistence of chemical residues in wool has stimulated interest in biological control of sheep lice by Bacillus thuringiensis.


The relation of insecure attachment states of mind and romantic attachment styles to adolescent aggression in romantic relationships.
The relation of attachment states of mind and self reported attachment relationship styles to romantic partner aggression was examined in a community sample of 93 adolescents. Higher levels of insecure-preoccupied and insecure-dismissing states of mind, as assessed by the Adolescent Attachment Interview at age 14, were predictive, respectively, of perpetration and victimization of psychological aggression in romantic relationships four years later. Partners' romantic attachment anxiety was linked to both psychological and physical aggression perpetration in romantic relationships. Results are interpreted as suggesting the value of assessing aggression in adolescent romantic relationships in the context of broader patterns of regulation of affect and behavior via the attachment system.


Strengthened tuberculosis control programme and trend of multi-drug resistant tuberculosis rate in Osaka City, Japan.
Osaka City has the highest tuberculosis (TB) notification rates in Japan. In the period 1999-2003, the TB control programme was strengthened, and the Stop TB Strategy was implemented to reduce the number of notified cases. The objective of this study was to assess the effect of these control activities in Osaka City, including the implementation of directly observed treatment (DOT), by analysing TB surveillance and routinely collected data. We reviewed the surveillance data of all sputum smear-positive pulmonary tuberculosis (PTB) cases registered in the Osaka City Public Health Office from 2001 to 2008 and data collected from the routine TB programme. The DOT implementation rate increased from 0% in 2001 to 68% in 2008 for smear-positive PTB cases of the general public and to 61% for all PTB cases of the homeless. The proportion of smear-positive PTB cases that had treatment failure and default combined, declined from 8.0% (52 of 650) in 2001 to 3.6% (20 of 548) in 2006. The proportion of cases among the homeless with previous treatment declined from 28% in 2001 to 15% in 2008. The proportion of cases with multidrug resistant-TB (MDR-TB) among those without previous treatment declined from 1.7% in 2001 to 0.9% in 2008. It is logical that reduction in the failure and default rate would lead to the reduction of cases with previous treatment and TB transmission, including resistant TB, therefore to the reduction of MDR-TB rates.


Perfluorooctane sulfonate exerts inflammatory bowel disease-like intestinal injury in rats.
Perfluorooctane sulfonate (PFOS), a type of perfluorinated compounds (PFCs), can induce various organ toxicity, including hepatomegaly, immunotoxicity, and gut microbiota disorder. PFCs have been associated with inflammatory bowel disease (IBD). Yet, whether PFOS exposure causes IBD-like disorder and the underlying mechanism remains undefined. Here, we investigated the influence of PFOS exposure on the development of IBD-like disorder in rats. Sprague-Dawley rats were intraperitoneally injected with PFOS (1 or 10 mg/kg) or normal saline (NS) every other day for 15 days. Body weight, serum concentrations of serum amyloid A (SAA) and high sensitivity C reactive protein (hsCRP) were measured. Pathological assessments of villi height and crypt depth in the proximal duodenum and jejunum were performed using H&E staining. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was used to assay cell apoptosis in the jejunum. The infiltration of inflammatory cells and cytokines in the jejunum were detected by immunohistochemistry analysis. PFOS (10 mg/kg) significantly increased the body weight, SAA and hsCRP, whereas no significant differences were observed in PFOS 1 mg/kg group of rats. The villi height and crypt depth in the proximal duodenum and jejunum were significantly reduced upon PFOS exposure. PFOS induced higher histopathological score in intestinal tissues compared to NS. Notably, TUNEL-positive cells were significantly higher in the jejunum upon PFOS exposure. Further, neutrophil and macrophage accumulated, and inflammatory cytokines infiltration were also remarkably increased in rats exposed to PFOS. PFOS induces IBD-like phenotypes in rats, with associated inflammatory infiltration to intestinal.


GFP-golvesin constructs to study Golgi tubulation and post-Golgi vesicle dynamics in phagocytosis.
Dictyostelium cells are professional phagocytes that are optimally suited for the imaging of phagosome processing from particle uptake to exocytosis. In order to design fluorescent probes for monitoring membrane trafficking in the endocytic pathway, we have dissected a membrane protein, golvesin, and have linked fragments of its sequence to GFP. Endogenous golvesin is partitioned between the ER, the Golgi apparatus, endosomes, and the contractile vacuole complex. We have localized signals that are required for exit from the Golgi to post-Golgi compartments to the C-terminal region of the golvesin sequence. One GFP-tagged fragment turned out to be a highly specific Golgi marker and was used to demonstrate the interaction of Golgi tubules with phagosomes. Signals essential for the retrieval of golvesin at the end of phagosome processing were localized to the N-terminal region. A truncated golvesin construct escaping retrieval was employed in recording the delivery of a phagosomal protein to the plasma membrane. Applying this construct to a phagosome filled with multiple particles, we observed that the phagosome is segmented during exocytosis, meaning that sequential release of particles alternates with membrane fusion.


Natural history and determinants of clinical severity of sickle cell disease.
Some of the factors determining the extremely variable clinical course of homozygous sickle cell disease are being identified. Genetic factors include alpha-thalassemia, beta-globin gene haplotypes, heterocellular hereditary persistence of fetal hemoglobin, and high total hemoglobin. Other factors include a variety of environmental variables and socioeconomic status. These risk factors, when occurring in conjunction with apparently random precipitating events, produce features of the disease. Many complications--which are age specific, with highest morbidity and mortality in the first 5 years--may be prevented by specific education and prophylaxis. Current median survival in individuals with sickle cell disease in the United States is approximately 40 to 50 years, at which age the major determinants of mortality are chronic end organ damage of the lungs and kidneys.


Randomized phase II chemotherapy and radiotherapy trial for patients with locally advanced inoperable non-small-cell lung cancer: long-term follow-up of RTOG 92-04.
The standard treatment for patients with locally advanced inoperable non-small-cell lung cancer and good prognostic factors has become combined chemotherapy (ChT) and radiotherapy (RT). However, the sequencing of the two modalities, as well as fractionation of RT, has been controversial. The Radiation Therapy Oncology Group (RTOG) Study 92-04 was a randomized Phase II study designed to evaluate further the toxicity and efficacy of 2 different strategies of chemoradiation evaluated in 2 prior RTOG Phase II studies. Patients with Stage II or III medically inoperable or unresectable non-small-cell lung cancer, good performance status, and minimal weight loss were enrolled into a prospective randomized Phase II RTOG study. Arm 1 consisted of induction ChT (vinblastine 5 mg/m(2) i.v. bolus weekly for the first 5 weeks, and cisplatin, 100 mg/m(2) i.v. on Days 1 and 29) followed by concurrent ChT/RT (cisplatin 75 mg/m(2) i.v. on Days 50, 71, and 92) during thoracic radiotherapy (63 Gy in 34 fractions during 7 weeks starting on Day 50). Arm 2 was concurrent ChT and hyperfractionated RT starting on Day 1 with a total dose of 69.6 Gy in 58 fractions during 6 weeks, 1.2 Gy/fraction b.i.d. ChT consisted of cisplatin, 50 mg/m(2) i.v. on Days 1 and 8, and oral VP-16, 50 mg b.i.d. for 10 days only on the days of thoracic radiotherapy repeated on Day 29. A total of 168 patients were entered between 1992 and 1994, and 163 patients were eligible for analysis. Eighty-one patients were treated in Arm 1 and 82 patients in Arm 2. Pretreatment characteristics, including age, gender, Karnofsky performance status, histologic features, and stage, were similar. The incidence of acute esophagitis was significantly higher among patients treated in Arm 2 than among those treated in Arm 1 (p <0.0001). The incidence of acute hematologic toxicity was significantly higher among patients treated in Arm 1 (p = 0.01 for anemia and p = 0.03 for other hematologic toxicities) than among those treated in Arm 2. Analysis of late toxicity showed that chronic esophageal toxicity was significantly more frequent in Arm 2 than in Arm 1 (p = 0.003). The time to in-field progression was significantly different (p = 0.009), favoring Arm 2 compared with Arm 1 (26% vs. 45% with failure in 2 years and 30% vs. 49% with failure in 4 years, respectively). The median 2-year and overall 5-year survival rates were similar between the two arms. Concurrent ChT and hyperfractionated RT resulted in a significant prolongation of the time to in-field progression, but with higher acute and chronic esophagitis. No other significant differences were observed between the two groups. Investigation with a chemoradio-protector is under way.


[Small interference RNA targeting nuclear factor-kappaB inhibits endometriotic angiogenesis in chick embryo chorioallantocic membrane].
To investigate the effect of small interference RNA (siRNA) targeting nuclear factor-kappaB (NF-kappaB) on endometriosis. The eutopic endometrium of women with endometriosis were transplanted into the nonvascular region of 8-day-old chicken embryo chorioallantocic membrane (CAM), and the effects of NF-kappaB p65 siRNA on the vascularization and endometriotic lesion formation were tested with proper controls. Transplantation of the endometrium onto the CAM resulted in a strong angiogenic response in the chicken tissue. The angiogenesis was significantly reduced and endometriotic lesion formation significantly suppressed with siRNA targeting NF-kappaB in comparison with the control group. The NF-kappaB pathway is involved in the development of endometriotic lesions in vitro, and NF-kappaB gene silencing reduces endometriotic angiogenesis and promotes cell apoptosis in the endometriotic lesions, suggesting that NF-kappaB might be a good target for endometriosis treatment.


Ortho-to-para ratio in interstellar water on the sightline toward Sagittarius B2(N).
The determination of the water ortho-to-para ratio (OPR) is of great interest for studies of the formation and thermal history of water ices in the interstellar medium and protoplanetary disk environments. We present new Herschel observations of the fundamental rotational transitions of ortho- and para-water on the sightline toward Sagittarius B2(N), which allow improved estimates of the measurement uncertainties due to instrumental effects and assumptions about the excitation of water molecules. These new measurements, suggesting a spin temperature of 24-32 K, confirm the earlier findings of an OPR below the high-temperature value on the nearby sightline toward Sagittarius B2(M). The exact implications of the low OPR in the galactic center molecular gas remain unclear and will greatly benefit from future laboratory measurements involving water freeze-out and evaporation processes under low-temperature conditions, similar to those present in the galactic interstellar medium. Given the specific conditions in the central region of the Milky Way, akin to those encountered in active Galactic nuclei, gas-phase processes under the influence of strong X-ray and cosmic ray ionization also have to be carefully considered. We summarize some of the latest laboratory measurements and their implications here.


Chronic helminth infection induces alternatively activated macrophages expressing high levels of CCR5 with low interleukin-12 production and Th2-biasing ability.
Helminth infections induce Th2-type biased immune responses. Although the mechanisms involved in this phenomenon are not yet clearly defined, antigen-presenting cells (APC) could play an important role in this process. Here, we have used peritoneal macrophages (F4/80+) recruited at different times after challenge with Taenia crassiceps as APC and tested their ability to regulate Th1/Th2 differentiation. Macrophages from acute infections produced high levels of interleukin-12 (IL-12) and nitric oxide (NO), paralleled with low levels of IL-6 and prostaglandin E(2) (PGE(2)) and with the ability to induce strong antigen-specific CD4+ T-cell proliferation in response to nonrelated antigens. In contrast, macrophages from chronic infections produced higher levels of IL-6 and PGE(2) and had suppressed production of IL-12 and NO, associated with a poor ability to induce antigen-specific proliferation in CD4+ T cells. Failure to induce proliferation was not due to a deficient expression of accessory molecules, since major histocompatibility complex class II, CD40, and B7-2 were up-regulated, together with CD23 and CCR5 as infection progressed. These macrophages from chronic infections were able to bias CD4+ T cells to produce IL-4 but not gamma interferon (IFN-gamma), contrary to macrophages from acute infections. Blockade of B7-2 and IL-6 and inhibition of PGE(2) failed to restore the proliferative response in CD4+ T cells. Furthermore, studies using STAT6(-/-) mice revealed that STAT6-mediated signaling was essential for the expansion of these alternatively activated macrophages. These data demonstrate that helminth infections can induce different macrophage populations that have Th2-biasing properties.


ANF system in the newborn piglet pulmonary vessels.
The atrial natriuretic factor (ANF) induces diuresis, natriuresis, and vasodilation. Although it was originally found to be secreted from the atria, ANF synthesis has been demonstrated in other organs. The adult lung is not only the first target organ for ANF, but it also expresses the ANF gene and synthesizes, releases, and clears ANF from the circulation. We have shown the presence of ANF in human fetal lungs and also demonstrated that these lungs can release bioactive ANF. However, the role of the ANF system in the newborn lung is unknown. Therefore we studied the ANF system in pulmonary vessels (arteries and veins dissected from the hilum down to a 100-microm diameter), in isolated perfused lungs, and in the plasma from pulmonary artery and vein of 1- and 7-day-old piglets. High-performance liquid chromatography (HPLC) revealed the presence of both the mature peptide and the ANF prohormone in pulmonary vein microsomes, but in pulmonary arteries, only the mature form was identified. Furthermore, in the veins, the ANF content tended to be higher in 7- than in 1-day-olds. ANF caused a dose-dependent decrease in perfusion pressure (p < 0.05). In veins and arteries, most of the ANF receptors were of the type A guanylate cyclase as opposed to clearance receptors. Interestingly, the ANF receptors were fewer in veins, where synthesis takes place, than in arteries (p < 0.05). Significant circulating ANF plasma levels were measured by radioimmunoassay in both pulmonary artery and vein. However, there was no site difference in ANF plasma levels, suggesting that ANF is cleared and synthesized in the pulmonary vessels. In conclusion, the entire ANF system is present in the newborn piglet pulmonary vessels. The paucity of clearance receptors compared with functional receptors potentiates the role of ANF in the regulation of postnatal pulmonary vascular resistance.


Gynecologic cancer among the socioeconomically disadvantaged.
It is difficult to discern the true dimensions of the relationship between poverty and gynecologic cancer. In well designed studies of patients with gynecologic cancers, demographic stratification usually is performed based on race/ethnicity, age, or geographic locale, but not on economic class. The unstated assumption of many of these reports is that women of color, inhabitants of rural communities, and older women are all poor. Although it is true that these populations are overrepresented among the poor, unless the variable of economic class is specifically evaluated, the broad nature of the problem may go unappreciated: the status of gynecologic cancers among the poor is primarily a reflection of a deeply rooted structural problem in the U.S. economy, the reverberations of which are experienced by all women who cannot afford regular health care. When women are poor and have gynecologic cancers, they often seek orthodox health care only after the symptoms have become unbearable. Explanations of this phenomenon include underlying feelings of pessimism, fatalism, or low self-esteem; faith in a belief system that does not regard the physician as the person to whom one goes for prevention or treatment of diseases; inaccessibility of health care facilities; experiences interpreted as degrading once health care facilities are accessed; high risk behavior and inability to pay. Programs that find effective ways around structural and functional problems of daily life and that respect and understand cultural norms have the best chance of finding temporary solutions to this national problem.


Requisite elements in vaccine immunity to Blastomyces dermatitidis: plasticity uncovers vaccine potential in immune-deficient hosts.
Understanding fundamental mechanisms of vaccine immunity will allow proper use and optimization of vaccines. Vaccination with a genetically engineered, live, attenuated strain of Blastomyces dermatitidis carrying a targeted deletion at the BAD1 locus confers sterilizing immunity against experimental lethal pulmonary infection. We found in this study that alphabeta T cells are requisite for durable vaccine immunity, whereas other T and B cells are dispensable. In immune-competent animals, CD4(+) T-cell derived cytokines TNF-alpha and IFN-gamma mediate vaccine immunity. Surprisingly, these factors are dispensable in immune-deficient animals, which rely on alternate mechanisms for robust vaccine immunity, yet still require O(2)(-) production rather than generation of NO. Our results clarify the cellular and molecular bases behind the first genetically engineered fungal vaccine. They also illustrate a sharp difference in vaccine mechanisms between immune-competent and immune-deficient hosts, which underscores the plasticity of residual immune elements in compromised hosts, and points to the feasibility of developing vaccines against invasive fungal infection in this fast growing patient population.


Inhibition of apoptosis signal-regulating kinase 1 mitigates the pathogenesis of human immunodeficiency virus-associated nephropathy.
Chronic kidney disease (CKD) is a common cause of morbidity and mortality in human immunodeficiency virus (HIV)-positive individuals. Among the HIV-related kidney diseases, HIV-associated nephropathy (HIVAN) is a rapidly progressive renal disease characterized by collapsing focal glomerulosclerosis (GS), microcystic tubular dilation, interstitial inflammation and fibrosis. Although the incidence of end-stage renal disease due to HIVAN has dramatically decreased with the widespread use of antiretroviral therapy, the prevalence of CKD continues to increase in HIV-positive individuals. Recent studies have highlighted the role of apoptosis signal-regulating kinase 1 (ASK1) in driving kidney disease progression through the activation of p38 mitogen-activated protein kinase and c-Jun N-terminal kinase and selective ASK-1 inhibitor GS-444217 was recently shown to reduce kidney injury and disease progression in various experimental models. Therefore we examined the efficacy of ASK1 antagonism by GS-444217 in the attenuation of HIVAN in Tg26 mice. GS-444217-supplemented rodent chow was administered in Tg26 mice at 4 weeks of age when mild GS and proteinuria were already established. After 6 weeks of treatment, the kidney function assessment and histological analyses were performed and compared between age- and gender-matched control Tg26 and GS-444217-treated Tg26 mice. GS-444217 attenuated the development of GS, podocyte loss, tubular injury, interstitial inflammation and renal fibrosis in Tg26 mice. These improvements were accompanied by a marked reduction in albuminuria and improved renal function. Taken together, GS-4442217 attenuated the full spectrum of HIVAN pathology in Tg26 mice. ASK1 signaling cascade is central to the development of HIVAN in Tg26 mice. Our results suggest that the select inhibition of ASK1 could be a potential adjunctive therapy for the treatment of HIVAN.


Glycosylation of recombinant ancrod from Agkistrodon rhodostoma after expression in mouse epithelial cells.
The thrombin-like serine protease ancrod from the Malayan pit viper Agkistrodon rhodostoma was expressed in mouse epithelial cells (C127). Oligosaccharide constituents were liberated from tryptic glycopeptides by treatment with peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidase F. Neutral oligosaccharide alditols obtained after reduction and enzymic desialylation were separated by two-dimensional HPLC and characterized by methylation analysis, liquid secondary-ion mass spectrometry, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and sequential degradation with exoglycosidases. In contrast to natural ancrod, the recombinant glycoprotein carries exclusively diantennary, triantennary and tetraantennary N-glycans with Gal beta 4 GlcNAc beta (type-2) antennae which were, in part, further substituted by host-cell-specific structural elements such as Gal alpha 3 residues or N-acetyllactosamine repeats. As a characteristic feature, a substantial proportion of the oligosaccharides bears a GalNAc beta 4Glc-NAc antenna. Studies at the level of individual N-glycosylation sites demonstrated that glycans with N, N'-diacetyllactosediamine units are not specifically attached but occur at all sites in varying amounts. Hence, the putative recognition signal (Pro70-Lys-Lys) for glycoprotein hormone N-acetylgalactosaminyltransferase, present in this glycoprotein in close proximity to Asn79, does not convey site-specific transfer of GalNAc residues in these cells.


[Preparation of highly-purified RNA from yeasts using calcium ions].
The influence of temperature and Ca2+ ions at different concentrations on the effectiveness of RNA precipitation from water-salt extract of baker yeast was studied. Our findings allowed us to propose a method for producing highly purified RNA by precipitation with 180 mM Ca2+ ions from hot yeast extracts (95 degrees C) after removal of the protein fraction that could be also precipitated with 20 mM Ca2+ at 90 degrees C. The purity and yields of RNA samples obtained in such a manner was of 98% and 56% relative to theoretical values, respectively.


Recreational training improves cardiovascular adaptations, metabolic profile and mental health of elderly women with type-2 diabetes mellitus.
We investigated the effects of a recreational training (RET) protocol in elderly women with type-2 diabetes mellitus (T2DM). We hypothesized that non-conventional physical activities of RET protocol might improve clinical outcomes regarding cardiovascular function, metabolic profile and mental health as participants keep the adherence to the protocol during the 3-month follow-up. Cardiovascular parameters (heart rate, systolic and diastolic blood pressure), circulating biomarkers (glucose and lipids) and salivary cortisol were attenuated in response to exercise. RET also reduced anxiety and depression indexes. RET protocol constitutes a potential therapeutic approach for managing T2DM in elderly women.


Initiation and regulation of water deficit-induced abscisic acid accumulation in maize leaves and roots: cellular volume and water relations.
Water deficit-induced ABA accumulation in relation to cellular water relations was investigated in maize root and leaf tissues. While polyethylene glycol (PEG) treatment led to a significant increase of ABA content in both root and leaf tissues, ethylene glycol (EG), a permeable monomer of PEG, had no effect on ABA accumulation at similar or much lower osmotic potentials. A rapid and massive accumulation of ABA in leaf tissues occurred at a specific threshold of PEG 6000 concentration, about 20% (w/v), and closely coincided with the start of the tissue weight loss and the obvious decrease of cellular osmotic potential. Pretreatment with EG lowered the cell sap osmotic potential and also lowered the capability of both root and leaf tissues to accumulate ABA in response to further air-drying or PEG treatment. When samples were dehydrated and incubated under pressure, a method to maintain high water potential and pressure potential during dehydration, ABA accumulation was similar to those dehydrated and incubated under atmospheric pressure. Such results suggest that both the absolute water potential and pressure potential per se had no direct effects on the dehydration-induced ABA accumulation. The results have provided evidence that the initiation of ABA accumulation is related to the weight loss of tissues or changes in cellular volume rather than the cell water relation parameters, and the capability of ABA accumulation can be regulated by cellular osmotic potential.


Effect of erythropoietin administration on proteins participating in iron homeostasis in Tmprss6-mutated mask mice.
Tmprss6-mutated mask mice display iron deficiency anemia and high expression of hepcidin. The aim of the study was to determine the effect of erythropoietin administration on proteins participating in the control of iron homeostasis in the liver and spleen in C57BL/6 and mask mice. Administration of erythropoietin for four days at 50 IU/mouse/day increased hemoglobin and hematocrit in C57BL/6 mice, no such increase was seen in mask mice. Erythropoietin administration decreased hepcidin expression in C57BL/6 mice, but not in mask mice. Erythropoietin treatment significantly increased the spleen size in both C57BL/6 and mask mice. Furthermore, erythropoietin administration increased splenic Fam132b, Fam132a and Tfr2 mRNA content. At the protein level, erythropoietin increased the amount of splenic erythroferrone and transferrin receptor 2 both in C57BL/6 and mask mice. Splenic ferroportin content was decreased in erythropoietin-treated mask mice in comparison with erythropoietin-treated C57BL/6 mice. In mask mice, the amount of liver hemojuvelin was decreased in comparison with C57BL/6 mice. The pattern of hemojuvelin cleavage was different between C57BL/6 and mask mice: In both groups, a main hemojuvelin band was detected at approximately 52 kDa; in C57BL/6 mice, a minor cleaved band was seen at 47 kDa. In mask mice, the 47 kDa band was absent, but additional minor bands were detected at approximately 45 kDa and 48 kDa. The results provide support for the interaction between TMPRSS6 and hemojuvelin in vivo; they also suggest that hemojuvelin could be cleaved by another as yet unknown protease in the absence of functional TMPRSS6. The lack of effect of erythropoietin on hepcidin expression in mask mice can not be explained by changes in erythroferrone synthesis, as splenic erythroferrone content increased after erythropoietin administration in both C57BL/6 and mask mice.


Speedy orthodontics: a case report.
Management of severe rotation poses a great challenge for the orthodontist, especially when cortical anchorage occurs. The conventional methods require excessive treatment time, rely on patient compliance, and can cause root resorption. A groundbreaking new procedure developed by Wilcko et al, which is actually a modification of the conventional corticotomy procedure, amalgamates the orthodontic mechanics, alveolar decortications, and augmentation procedure to make treatment time three to four times faster than conventional orthodontic techniques. This procedure uses the dynamics of bone physiology and redirects the emphasis in tooth movement to the manner in which supporting bone responds to orthodontic forces applied to the tooth. This article includes a case report in which the technique is used on a patient who has a severely rotated mandibular left canine.


Determination of Dibromochloropropane and Related Fumigants in Citrus Fruit.
A simple and systematic procedure for determination of 1,2-dibromo-3-chloropropane (DBCP), 1,3-dichloropropene (DCP), chlorobutol (TCB) and seven related compounds in citrus fruits was established. Since these compounds have extremely high volatility, the distillation apparatus for volatile oil determination was used. These compounds were trapped into cyclohexane (more than 94.2-98.4% at spiking levels of 0.5-50 ppm) by this distillation. The distillate was purified by Florisil column chromatography. All compounds except TCB were eluted with n-hexane:ether(93:7) and TCB was eluted with ether. Each eluate was injected into an ECD-gas chromatograph directly for determination. Their recoveries from lemon, orange and grapefruit were within 91.5-98.6% at the spiking levels of 0.01-1 ppm.


Histopathology of tick-bite lesions in naturally infested capybaras (Hydrochoerus hydrochaeris) in Brazil.
In the present work features of tick-bite lesions were evaluated in capybaras naturally infested with Amblyomma cajennense and Amblyomma dubitatum ticks. Gross appearance of tick bite site was characterized by a mild swelling and erythema. Microscopic examination revealed the cement cone, a tube-like homogenous eosinophilic mass penetrating deep into the dermis. This structure was surrounded in the dermis by a cellular infiltrate and free eosinophilic granules and was associated to edema of variable intensity. Necrosis was a common feature deep in the dermis particularly at the far end of the eosinophilic tube. Hyperplasia, cellular edema and occasionally necrosis of keratinocytes could be seen at both sides of the ruptured epidermis. Cellular infiltrate was constituted overwhelmingly by polymorphonuclear leukocytes with eosinophilic granules. In capybaras cells with such features can be either eosinophils or heterophils (pseudoeosinophils), the latter being the equivalent of neutrophils of other mammals. Ultrastructural analysis of the cellular infiltrate revealed the predominance of heterophils over eosinophils. Mononuclear cells and mast cells and, in lesser numbers, basophils were also seen at skin attachment sites. The presence of heterophils in the reaction of capybaras against Amblyomma ticks is an outstanding feature but its role in the reaction to the tick is not known. It is however speculated that capybara heterophils might be associated with a more permissive environment for tick feeding and pathogen transmission as already shown for the equivalent cell type, the neutrophil, in the reaction of the dog against the Rhipicephalus sanguineus tick.


Change in rehabilitation needs and activity limitations over time of adults with acquired visual impairment following entry to a low vision rehabilitation service in England.
Longitudinal changes in priority rehabilitation needs, vision-related activity limitation and importance of visual goals were evaluated in a sample of people with a visual impairment over a year following entry to low vision rehabilitation services in England. Participants were adults with newly registered visual impairment within Leicestershire. Priority scores, indicating the level of rehabilitative need, were determined from the importance and difficulty scores of the 48 goals of the Participation and Activity Inventory (PAI). Rasch analysis of the difficulty and importance scores examined activity limitation and importance separately. PAI outcome measures were assessed on entry to rehabilitation services and at 4 and 12 months thereafter. Forty-eight participants (mean age 74.2, SD 14.1 years) completed three visits. Overall, there was a statistically significant reduction in the perceived need for rehabilitation over time (p < 0.001, ηp2 = 0.29), driven by reduced perceived difficulty (p < 0.001, ηp2 = 0.32) but stable importance (p = 0.73) of goals, with most change occurring between baseline and 4 months. PAI goals with greatest rehabilitative need at study entry were reading, mobility and writing, and these remained of highest priority over time. The greatest priority score decrease was for the goal 'Hobbies and crafts'. The largest decrease in importance was for 'Mobility outdoors', whereas 'Relationship with loved ones' increased most. Despite a decline in the perceived need for rehabilitation over the study period, there remains a need for continued support and intervention at 12 months following registration with rehabilitation services, particularly for the key goals of reading, writing and mobility. Early identification and support for individuals' important but difficult goals could prevent such goals being relinquished. Goals concerning relationships and communication became more important over time, indicating that re-evaluation of needs at follow-up is necessary to inform ongoing service provision.


Evolution of an atypical de-epoxidase for photoprotection in the green lineage.
Plants, algae and cyanobacteria need to regulate photosynthetic light harvesting in response to the constantly changing light environment. Rapid adjustments are required to maintain fitness because of a trade-off between efficient solar energy conversion and photoprotection. The xanthophyll cycle, in which the carotenoid pigment violaxanthin is reversibly converted into zeaxanthin, is ubiquitous among green algae and plants and is necessary for the regulation of light harvesting, protection from oxidative stress and adaptation to different light conditions(1,2). Violaxanthin de-epoxidase (VDE) is the key enzyme responsible for zeaxanthin synthesis from violaxanthin under excess light. Here we show that the Chlorophycean VDE (CVDE) gene from the model green alga Chlamydomonas reinhardtii encodes an atypical VDE. This protein is not homologous to the VDE found in plants and is instead related to a lycopene cyclase from photosynthetic bacteria(3). Unlike the plant-type VDE that is located in the thylakoid lumen, the Chlamydomonas CVDE protein is located on the stromal side of the thylakoid membrane. Phylogenetic analysis suggests that CVDE evolved from an ancient de-epoxidase that was present in the common ancestor of green algae and plants, providing evidence of unexpected diversity in photoprotection in the green lineage.


TRIPS, the Doha Declaration and increasing access to medicines: policy options for Ghana.
There are acute disparities in pharmaceutical access between developing and industrialized countries. Developing countries make up approximately 80% of the world's population but only represent approximately 20% of global pharmaceutical consumption. Among the many barriers to drug access are the potential consequences of the Trade Related Aspects of Intellectual Property Rights (TRIPS) Agreement. Many developing countries have recently modified their patent laws to conform to the TRIPS standards, given the 2005 deadline for developing countries. Safeguards to protect public health have been incorporated into the TRIPS Agreement; however, in practice governments may be reluctant to exercise such rights given concern about the international trade and political ramifications. The Doha Declaration and the recent Decision on the Implementation of Paragraph 6 of the Doha Declaration on the TRIPS Agreement and Public Health may provide more freedom for developing countries in using these safeguards. This paper focuses on Ghana, a developing country that recently changed its patent laws to conform to TRIPS standards. We examine Ghana's patent law changes in the context of the Doha Declaration and assess their meaning for access to drugs of its population. We discuss new and existing barriers, as well as possible solutions, to provide policy-makers with lessons learned from the Ghanaian experience.


Computational subunits of visual cortical neurons revealed by artificial neural networks.
A crucial step toward understanding visual processing is to obtain a comprehensive description of the relationship between visual stimuli and neuronal responses. Many neurons in the visual cortex exhibit nonlinear responses, making it difficult to characterize their stimulus-response relationships. Here, we recorded the responses of primary visual cortical neurons of the cat to spatiotemporal random-bar stimuli and trained artificial neural networks to predict the response of each neuron. The random initial connections in the networks consistently converged to regular patterns. Analyses of these connection patterns showed that the response of each complex cell to the random-bar stimuli could be well approximated by the sum of a small number of subunits resembling simple cells. The direction selectivity of each complex cell measured with drifting gratings was also well predicted by the combination of these subunits, indicating the generality of the model. These results are consistent with a simple functional model for complex cells and demonstrate the usefulness of the neural network method for revealing the stimulus-response transformations of nonlinear neurons.


Reduction of auxin transport capacity with age and internal water deficits in cotton petioles.
Auxin transport was examined in leaf petioles taken from the upper, middle, and lower leaf canopy of large cotton plants. The ability of petioles to transport auxin decreased with age (position) of the leaves. Plant water deficit reduced transport regardless of age. These correlations support the view that reduced transport capacity of petioles plays a significant role in the induction of abscission of lower or older leaves during water deficits.


Paroxetine increases steady-state concentrations of (R)-methadone in CYP2D6 extensive but not poor metabolizers.
Steady-state blood concentrations of (R)- methadone (i.e., the active form), (S)-methadone, and (R,S)-methadone were measured before and after introduction of paroxetine 20 mg/day during a mean period of 12 days in 10 addict patients in methadone maintenance treatment. Eight patients were genotyped as CYP2D6 homozygous extensive metabolizers (EMs) and two patients as poor metabolizers (PMs). Paroxetine significantly increased concentrations of both enantiomers of methadone in the whole group (mean increase for (R)-methadone +/- SD, 26 +/- 32%; range, -14% to +83%, p = 0.032; for (S)-methadone, 49 +/- 51%; range, -29% to +137%, p = 0.028; for (R,S)-methadone, 35 +/- 41%; range, -20% to +112%, p = 0.032) and in the group of eight EMs (mean increase, 32%, p = 0.036; 53%, p = 0.028; and 42%, p = 0.036, for (R)-methadone, (S)-methadone, and (R,S)-methadone, respectively). On the other hand, in the two PMs, (S)-methadone but not (R)-methadone concentrations were increased by paroxetine (mean increases of 36% and 3%, respectively). Paroxetine is a strong CYP2D6 inhibitor, and these results confirm previous studies showing an involvement of CYP2D6 in methadone metabolism with a stereoselectivity toward the (R)-enantiomer. Because paroxetine is a mild inhibitor of CYP1A2, CYP2C9, CYP2C19, and CYP3A4, increase of (S)-methadone concentrations in both EMs and PMs could be mediated by inhibition of any of these isozymes.


Intrauterine growth restriction in rats is associated with hypertension and renal dysfunction in adulthood.
Epidemiological studies have produced evidence that unfavorable intrauterine environments during fetal life may lead to adverse outcomes in adulthood. We have previously shown that a low-sodium diet, given to pregnant rats over the last week of gestation, results in intrauterine growth restriction (IUGR). We hypothesize that pups born with IUGR are more susceptible to the development of hypertension in adulthood. IUGR fetuses and rats aged 1 wk were characterized for organ growth and renal morphogenesis. The adults (12 wk) were evaluated for weight, systolic blood pressure, activity of the renin-angiotensin-aldosterone system (RAAS), and renal function; hearts and kidneys underwent a histological examination. Brain and cardiac ventricle-to-body ratios were increased in IUGR fetuses compared with age-matched controls, whereas the kidney-to-body ratio was unchanged. Systolic blood pressure was elevated in both IUGR male and female adults. Plasma aldosterone levels were not correlated with increased plasma renin activity. Moreover, urinary sodium was decreased, whereas plasma urea was elevated in both males and females, and creatinine levels were augmented only in females, suggesting a glomerular filtration impairment in IUGR. In our model of IUGR induced by a low-sodium diet given to pregnant rats, high blood pressure, alteration of the RAAS, and renal dysfunction are observed in adult life. Differences observed between male and female adults suggest the importance of gender in outcomes in adulthood after IUGR.


Note: susceptibility to chlorine of Aeromonas hydrophila strains.
The susceptibility of five Aeromonas hydrophila strains and one Escherichia coli strain to chlorine was studied under carefully controlled laboratory conditions. Of the Aer. hydrophila strains, two were from untreated water, two from tap water (immediately downstream of a water treatment plant) and one from the DSM collection. The study included disinfectant concentration (0.1, 0.2 and 0.5 mg l-1), pH (6, 7 and 8) and temperature (4, 21 and 32 degrees C) as controlled variables. The results indicated that the untreated water strains, the DSM strain and the E. coli strain were inactivated within 1 min of chlorine treatment. The strains from chlorinated water (TW11 and TW27) showed a different susceptibility to chlorine disinfection, the rate of inactivation being greater at pH6 than at pH8 for both strains. Under the standard conditions of temperature 21 degrees C, pH7 and chlorine concentration 0.2 mg l-1, an increase or decrease of approximately 1 log unit in the number of bacteria did not affect the kill rate of the strains TW11 and TW27.


[Multiple primary neoplasms on the face].
A case of 74-year old woman with six separate cancers is presented. Four of them were nonmelanoma skin cancers (2 basal-cell carcinomas and 2 squamous-cell carcinomas), the remaining two were noncutaneous malignancies (invasive lobular carcinoma of the breast and squamous cell carcinoma of the parotid gland). Three lesions were present simultaneously on the head and neck region; basal-cell carcinoma on the cheek, squamous-cell carcinomas on the forehead, and in the parotid gland. All neoplasms were surgically removed, the region of affected parotid gland was additionally radiated. Extreme rarity of the multifocal primary malignancies is emphasized and the non-completely clear etiology, biology and pathology of these tumors is discussed.


Review of the Isolation, Characterization, Biological Function, and Multifarious Therapeutic Approaches of Exosomes.
Exosomes are extracellular vesicles that contain a specific composition of proteins, lipids, RNA, and DNA. They are derived from endocytic membranes and can transfer signals to recipient cells, thus mediating a novel mechanism of cell-to-cell communication. They are also thought to be involved in cellular waste disposal. Exosomes play significant roles in various biological functions, including the transfer of biomolecules such as RNA, proteins, enzymes, and lipids and the regulation of numerous physiological and pathological processes in various diseases. Because of these properties, they are considered to be promising biomarkers for the diagnosis and prognosis of various diseases and may contribute to the development of minimally invasive diagnostics and next generation therapies. The biocompatible nature of exosomes could enhance the stability and efficacy of imaging probes and therapeutics. Due to their potential use in clinical applications, exosomes have attracted much research attention on their roles in health and disease. To explore the use of exosomes in the biomedical arena, it is essential that the basic molecular mechanisms behind the transport and function of these vesicles are well-understood. Herein, we discuss the history, biogenesis, release, isolation, characterization, and biological functions of exosomes, as well as the factors influencing their biogenesis and their technical and biological challenges. We conclude this review with a discussion on the future perspectives of exosomes.


Dealing with immunogenicity of biologicals: assessment and clinical relevance.
In the last decade, biologicals revolutionized rheumatology. An increasing number of patients benefit from biotherapeuticals. However, some patients do not respond to treatment and others lose their response after a certain time. Immunogenicity is one of the factors linked to secondary nonresponse but its clinical significance has remained controversial. In recent years, knowledge of how to assess immunogenicity of biologicals has improved. Various reports show an inverse relationship between drug levels and antibody formation against the drug. Studies associated immunogenicity of therapeutic antibodies with clinically significant nonresponse in a subgroup of patients. Clinically relevant immunogenicity is influenced by several factors including dosing and concomitant medication. It has been shown that immunogenicity against biologicals can be persistent or transient. Immunogenicity affects a significant number of patients treated with biologicals. Monitoring of drug levels as well as of antibodies against therapeutic antibodies may lead to more rational treatment strategies.


Prediction of protein-protein interactions between Helicobacter pylori and a human host.
A lack of information on protein-protein interactions at the host-pathogen interface is impeding the understanding of the pathogenesis process. A recently developed, homology search-based method to predict protein-protein interactions is applied to the gastric pathogen, Helicobacter pylori to predict the interactions between proteins of H. pylori and human proteins in vitro. Many of the predicted interactions could potentially occur between the pathogen and its human host during pathogenesis as we focused mainly on the H. pylori proteins that have a transmembrane region or are encoded in the pathogenic island and those which are known to be secreted into the human host. By applying the homology search approach to protein-protein interaction databases DIP and iPfam, we could predict in vitro interactions for a total of 623 H. pylori proteins with 6559 human proteins. The predicted interactions include 549 hypothetical proteins of as yet unknown function encoded in the H. pylori genome and 13 experimentally verified secreted proteins. We have recognized 833 interactions involving the extracellular domains of transmembrane proteins of H. pylori. Structural analysis of some of the examples reveals that the interaction predicted by us is consistent with the structural compatibility of binding partners. Examples of interactions with discernible biological relevance are discussed.


Defective function of GABA-containing synaptic vesicles in mice lacking the AP-3B clathrin adaptor.
AP-3 is a member of the adaptor protein (AP) complex family that regulates the vesicular transport of cargo proteins in the secretory and endocytic pathways. There are two isoforms of AP-3: the ubiquitously expressed AP-3A and the neuron-specific AP-3B. Although the physiological role of AP-3A has recently been elucidated, that of AP-3B remains unsolved. To address this question, we generated mice lacking mu3B, a subunit of AP-3B. mu3B-/- mice suffered from spontaneous epileptic seizures. Morphological abnormalities were observed at synapses in these mice. Biochemical studies demonstrated the impairment of gamma-aminobutyric acid (GABA) release because of, at least in part, the reduction of vesicular GABA transporter in mu3B-/- mice. This facilitated the induction of long-term potentiation in the hippocampus and the abnormal propagation of neuronal excitability via the temporoammonic pathway. Thus, AP-3B plays a critical role in the normal formation and function of a subset of synaptic vesicles. This work adds a new aspect to the pathogenesis of epilepsy.


Limited genetic polymorphism of the Plasmodium vivax low molecular weight rhoptry protein complex in the Colombian population.
Proteins involved in parasite adhesion and invasion are considered the best candidates for the development of asexual blood-stage antimalarial vaccines. Such vaccine candidates should be accessible by the immune system and have limited diversity. Considering the promising results obtained in previous trials by immunizing monkeys with the rhoptry-associated proteins 1 and 2 (RAP-1 and RAP-2), here we assessed the genetic variability of the Plasmodium vivax rap-1 and rap-2 genes isolated from Colombian parasite populations. Limited sequence diversity was found in these genes, possibly as a result of a functional/structural restriction. The presence of several haplotypes at relatively low frequencies and the excess of singleton mutations suggests that a demographic process might be affecting the loci. Our results support the inclusion of PvRAP-1 and PvRAP-2 in the design of an antimalarial subunit-based vaccine against P. vivax, which would avoid inducing allele-specific immunity.


Nature gives us strength: exposure to nature counteracts ego-depletion.
Previous research rarely investigated the role of physical environment in counteracting ego-depletion. In the present research, we hypothesized that exposure to natural environment counteracts ego-depletion. Three experiments were conducted to test this hypothesis. In Experiment 1, initially depleted participants who viewed pictures of nature scenes showed greater persistence on a subsequent anagram task than those who were given a rest period. Experiment 2 expanded upon this finding by showing that natural environment enhanced logical reasoning performance after ego-depleting task. Experiment 3 adopted a two- (depletion vs. no-depletion) -by-two (nature exposure vs. urban exposure) factorial design. We found that nature exposure moderated the effect of depletion on anagram task performance. Taken together, the present studies offer a viable and novel strategy to mitigate the negative impacts of ego-depletion.


Oxidative stress and Kawasaki disease: how is oxidative stress involved from the acute stage to the chronic stage?
Inflammation and oxidative stress are closely related. Further, oxidative stress plays an important role in the pathology of inflammation-based Kawasaki disease. An excessive in vivo production of reactive oxygen species increases oxidative stress in the body, which triggers an endless vicious spiral of inflammation reactions and reactive oxygen metabolites. This presumably forms diffuse vasculitis in the acute phase. Acute inflammation and oxidative stress can be rapidly controlled by treatments; however, they may remain for a long time. This has recently been identified as a problem in the chronic phase of Kawasaki disease. Generally, the presence of vascular inflammation and oxidative stress impairs blood vessels, leading to the onset of atherosclerosis, which is a widely recognized risk. The current discussion focuses on whether the same is valid for blood vessels in the chronic phase of Kawasaki disease.


Association of specific microorganisms with endodontic signs and symptoms. A comparative study.
The study aimed to evaluate the association of root canal microorganisms red complex and E. facealis with endodontic clinical signs and symptoms using polymerase chain reaction. Bacterial samples were obtained using sterile paper points from the teeth of 100 subjects divided into two groups; Group I: 50 individuals with primary tooth infections and Group II: 50 individuals with failed endodontic treatment having the secondary infection. DNA extracted from samples was analyzed for endodontic pathogens by using species-specific primers. The pain was noticed in 66%, (33 of 50 subjects) in primary infection and 60% (30 of 50) in a secondary infection. A statistically significant association between pain and E. faecalis bacteria observed both in primary infection and secondary infection (P < 0.05). Tenderness on percussion was associated with 40% cases in Group I and 70% cases in Group II. The red complex accounted for 94% of cases associated with tenderness on percussion in primary infection while 86% of cases associated with secondary infection with a statistically significant association (P < 0.05). Prevalence of red complex bacteria and E. faecalis suggested the association of studied bacteria with symptomatic infected pulp and periradicular diseases.


A multitechnique study of europium decatungstate and human serum albumin molecular interaction.
Polyoxometalates (POMs) show promising biological activities, but the mechanism of these potential therapeutic effects remains to be elucidated at a molecular level. As a step in this direction, the interaction between the Eu-containing decatungstate [EuW(10)O(36)](9-) and human serum albumin (HSA) has been studied by several techniques. Fluorescence/luminescence analysis showed the existence of a strong interaction between the POM and HSA. This interaction has key effects both on luminescence of the POM and on the behaviours of HSA. An enhancement of the POM luminescence is observed upon interaction. The presence of increasing concentrations of the POM results in the progressive quenching of the fluorescence of the single tryptophan of HSA. Circular dichroism led to the conclusion that the binding of the POM did not alter the secondary structure of HSA. Isothermal titration calorimetry revealed an enthalpy-driven binding reaction between HSA and the POM, resulting in the formation of a 1:1 complex. The present work is meaningful in finding novel solid state bio-image or fluorescence/luminescence labelling agents.


Specific demonstration of drug-induced tumour cell apoptosis in human xenografts models using a plasma biomarker.
Pharmacodynamic (PD) assays should be used before advancing new drugs to clinical trials. Most PD assays measure the response to drugs in tissue, a procedure which requires tissue biopsies. The M30-Apoptosense ELISA is a PD biomarker assay for the quantitative determination of caspase-cleaved cytokeratin 18 (CK18) released from apoptotic carcinoma cells into blood. We here demonstrate that whereas the M30-Apoptosense ELISA assay detects human caspase-cleaved CK18, the mouse and rat CK18 caspase cleavage products are detected with low affinity. The M30-Apoptosense ELISA therefore facilitates the determination of drug-induced apoptosis in human tumour xenografts in rodents using plasma samples, largely independently from host toxicity. Increases of caspase-cleaved CK18 were observed in plasma from different carcinoma xenograft models in response to anticancer drugs. The appearance caspase-cleaved CK18 in plasma was found to reflect formation of the caspase-cleaved epitope in FaDu head-neck carcinomas and in cultured cells. The M30-Apoptosense assay allows determination of tumour response in blood from xenograft models and from patients, providing a powerful tool for translational studies of anticancer drugs.


Role of the D1A dopamine receptor in the pathogenesis of genetic hypertension.
Since dopamine produced by the kidney is an intrarenal regulator of sodium transport, an abnormality of the dopaminergic system may be important in the pathogenesis of hypertension. In the spontaneously hypertensive rat (SHR), in spite of normal renal production of dopamine and receptor density, there is defective transduction of the D1 receptor signal in renal proximal tubules, resulting in decreased inhibition of sodium transport (Na+/H+ exchanger [NHE] and Na+/K+ATPase activity) by dopamine. To determine if impaired D1 receptor regulation of NHE in proximal tubules is related to hypertension, studies were performed in a F2 generation from female Wistar Kyoto (WKY) and male SHR crosses. A D1 agonist, SKF 81297, inhibited (37.6 +/- 4.7%) NHE activity in brush border membranes of normotensive F2s (systolic blood pressure < 140 mm Hg, n = 7) but not in hypertensive F2s (n = 21). Furthermore, a D1 agonist, SKF 38393, when infused into the renal artery, dose dependently increased sodium excretion in normotensive F2s (n = 3) without altering renal blood flow but was inactive in hypertensive F2s (n = 21). Since the major D1 receptor gene expressed in renal proximal tubules is the D1A subtype, we determined the importance of this gene in the control of blood pressure in mice lacking functional D1A receptors. Systolic blood pressure was greater in homozygous (n = 6) and heterozygous (n = 5) mice compared to normal sex matched litter mate controls (n = 12); moreover, the mice lacking one or both D1A alleles developed diastolic hypertension. The cosegregation with hypertension of an impaired D1 receptor regulation of renal sodium transport and the development of elevated systolic and diastolic pressure in mice lacking one or both D1A alleles suggest a causal relationship of the D1A receptor gene with hypertension.