A proline-rich region with a highly periodic sequence in Streptococcal beta protein adopts the polyproline II structure and is exposed on the bacterial surface.
Proline-rich regions have been identified in many surface proteins of pathogenic streptococci and staphylococci. These regions have been suggested to be located in cell wall-spanning domains and/or to be required for surface expression of the protein. Because little is known about these regions, which are found in extensively studied and biologically important surface proteins, we characterized the proline-rich region in one such protein, the beta protein of group B streptococci. The proline-rich region in beta, designated the XPZ region, has a proline at every third position, and the sequence is highly periodic in other respects. Immunochemical analysis showed that the XPZ region was not associated with the cell wall but was exposed on the bacterial surface. Moreover, characterization of a beta mutant lacking the XPZ region demonstrated that this region was not required for surface expression of the beta protein. Comparison of the XPZ region in different beta proteins showed that it varied in size but always retained the typical sequence periodicity. Circular dichroism spectroscopy indicated that the XPZ region had the structure of a polyproline II helix, an extended and solvent-exposed structure with exactly three residues per turn. Because of the three-residue sequence periodicity in the XPZ region, it is expected to be amphipathic and to have distinct nonpolar and polar surfaces. This study identified a proline-rich structure with unique properties that is exposed on the surface of an important human pathogen.


Life and death of the people of London: a historical GIS of Charles Booth's inquiry.
Social reformer Charles Booth undertook a massive survey into the social and economic conditions of the people of London at the end of the 19th century. An important innovation of his Inquiry was the construction of large, detailed maps displaying social class of inner London on a street-by-street basis. These provide a detailed and vivid picture of the geography of poverty and affluence at this time. These maps have been digitised, georeferenced and linked to contemporary ward boundaries allowing Booth's measurement of social class to be matched to the measurement of social class in the 1991 census of population and standardised mortality ratios derived for all causes of death in the survey area between 1991 and 1995. The social class data were used to derive an index of relative poverty for both time periods and a comparison of the geographies of relative poverty and their relationship with contemporary mortality was made. Although the overall standard of living had increased, the geography of poverty at the end of the 19th century was very similar to that at the end of the 20th century. Moreover, the geography of all causes of death for people over the age of 65 was more strongly related to the geography of poverty in the late 19th century than contemporary patterns of poverty. This relationship was also true for mortality for specific diseases that are related to deprivation in early life. The paper concludes that the spatial patterns of poverty in inner London are extremely robust and a century of change has failed to disrupt it.


Inhibition of Aerobic Glycolysis Attenuates Disease Progression in Polycystic Kidney Disease.
Dysregulated signaling cascades alter energy metabolism and promote cell proliferation and cyst expansion in polycystic kidney disease (PKD). Here we tested whether metabolic reprogramming towards aerobic glycolysis ("Warburg effect") plays a pathogenic role in male heterozygous Han:SPRD rats (Cy/+), a chronic progressive model of PKD. Using microarray analysis and qPCR, we found an upregulation of genes involved in glycolysis (Hk1, Hk2, Ldha) and a downregulation of genes involved in gluconeogenesis (G6pc, Lbp1) in cystic kidneys of Cy/+ rats compared with wild-type (+/+) rats. We then tested the effect of inhibiting glycolysis with 2-deoxyglucose (2DG) on renal functional loss and cyst progression in 5-week-old male Cy/+ rats. Treatment with 2DG (500 mg/kg/day) for 5 weeks resulted in significantly lower kidney weights (-27%) and 2-kidney/total-body-weight ratios (-20%) and decreased renal cyst index (-48%) compared with vehicle treatment. Cy/+ rats treated with 2DG also showed higher clearances of creatinine (1.98±0.67 vs 1.41±0.37 ml/min), BUN (0.69±0.26 vs 0.40±0.10 ml/min) and uric acid (0.38±0.20 vs 0.21±0.10 ml/min), and reduced albuminuria. Immunoblotting analysis of kidney tissues harvested from 2DG-treated Cy/+ rats showed increased phosphorylation of AMPK-α, a negative regulator of mTOR, and restoration of ERK signaling. Assessment of Ki-67 staining indicated that 2DG limits cyst progression through inhibition of epithelial cell proliferation. Taken together, our results show that targeting the glycolytic pathway may represent a promising therapeutic strategy to control cyst growth in PKD.


Depressed outpatients treated with cognitive therapy or pharmacotherapy. A one-year follow-up.
Using a controlled, clinical-trial format, 44 nonpsychotic, nonbipolar, depressed outpatients were treated with cognitive therapy or imipramine hydrochloride over a 12-week period. Although both interventions were associated with significant reductions in levels of depression, the cognitive-therapy patients showed greater symptomatic improvement and a higher treatment-completion rate. A one-year naturalistic follow-up of the 35 subjects who completed the protocol revealed that although many of the patients had a variable clinical course, both original treatment groups remained generally well. Self-rated depressive symptomatology was significantly lower for those who, one year earlier, had completed cognitive therapy than for those who had been in the clinical trial's pharmacotherapy cell. While there were several other interesting trends in favor of the cognitive-therapy patients, none of the between-group differences were significant. The pragmatic and clinical implications of the followup results are discussed.


Differences in chronic spontaneous urticaria between Europe and Central/South America: results of the multi-center real world AWARE study.
Global chronic urticaria (CU) disease experience and management is not well documented. This study descriptively compares these aspects among CU patients residing in Europe (EU) and Central and South America (C/SA). AWARE (A World-wide Antihistamine-Refractory chronic urticaria patient Evaluation) is a global prospective, non-interventional study of CU in the real-world setting. Patients were ≥ 18 years with a diagnosis of H1-antihistamine-refractory CU for > 2 months. Differences between the EU and C/SA regions in demographic and clinical characteristics, quality of life (QoL), work and activity impairment, pharmacological treatment, and healthcare resource use were examined. In total, 4224 patients were included in the analysis (C/SA 492; EU 3732). Rates of untreated patients were greater in the C/SA region (45.1% vs. 31.9%; P < 0.005) and escalation to third-line therapy was rare in both regions. Differences in disease experience emerged, with C/SA patients more commonly experiencing angioedema (C/SA 50.8% vs. EU 46.1%; P = 0.03) or comorbid chronic inducible urticaria (C/SA 30% vs. EU 22%; P < 0.001). Correspondingly, rates of uncontrolled urticaria were higher among C/SA patients (82.8% vs. 77.5%; P = 0.017) and patients in the C/SA region showed significantly greater work and activity impairment (absenteeism: 10.4 ± 19.7 vs. 6.7 ± 19.0, P = 0.004; presenteeism: 30.3 ± 31.9 vs. 24.4 ± 25.8, P = 0.001; work productivity loss: 33.9 ± 33.9 vs. 26.5 ± 27.5, P < 0.001; activity impairment: 37.7 ± 34.7 vs. 32.7 ± 30.1, P = 0.001). However, QoL impairment was greater in the EU region (Dermatology Life Quality Index: C/SA 6.5 ± 5.9 vs. EU 8.3 ± 7.0; P < 0.001). There was a significant difference in use of healthcare resources, including emergency services (39.6% vs. 29.3%; P < 0.001), hospitalization (7.7% vs 21.9%; P < 0.001) general practitioners (31.7% vs 57.3%; P < 0.001), and additional allergists or dermatologists (50.6% vs. 47.3%, P < 0.001), among patients in the C/SA and EU region, respectively. In both regions, patients with a primary diagnosis of CU with angioedema had significantly greater impairment in work and non-work activities and healthcare resource utilization compared to those without angioedema. This study revealed that CU is a heterogeneous condition with differences in healthcare utilization and outcomes between EU and C/SA. However, overall there is a high unmet need of H1-antihistamine-refractory CU patients, which is associated with high use of healthcare resources, and has a large negative effect on QoL and work productivity.


Self-reported treatment impressions and satisfaction of papulopustular rosacea patients treated with doxycycline, USP, 40 mg capsules.
This survey program was designed to evaluate patients' experiences with doxycycline, USP, 40 mg capsules (30 mg immediate release, 10 mg delayed release beads, ORACEA®; Galderma Laboratories, L.P.) as a treatment for the inflammatory lesions of rosacea and to provide patient-reported feedback to physicians. This prospective, cross-sectional survey was implemented in January 2010. One thousand, two hundred and ninety-five physicians identified patients eligible for treatment with doxycycline, USP, 40 mg capsules and provided them with information about the program. Patients voluntarily participated by contacting a program coordinator or by enrolling online, providing consent, and responding to a series of questions prior to medication use and approximately four weeks post-treatment initiation. Surveys were completed through an automated interactive voice response system or a dedicated, secure website and included questions regarding patients' perceptions of when the treatment first started working, patients' symptom severity, interference of symptoms with work and social activities, and confidence in appearance. Patients were also asked about prior medication use, adjunct medication use, and treatment satisfaction. Reports of patient progress and responses to these survey questions were sent directly to each patient's treating physician within a few days of survey completion. Paired t-tests were used to evaluate the statistical significance of differences in symptom severity, interference and confidence ratings before and after treatment. Two thousand, eight hundred ninety-eight patients enrolled in the survey program and completed the baseline survey. Of these, a total of 1,346 patients completed the baseline and four-week survey (mean age 50 years; 75% female). Most (58%) reported use of a prior prescription medication to treat rosacea. Over half the patients (52%) responded that the product began to work within two weeks of use. After four weeks of using doxycycline, USP, 40 mg capsules, patients felt that the severities of redness as well as bumps/blemishes were significantly reduced (P<.05). Patients also reported having more confidence with their skin's appearance (P<.05). In addition, with use of doxycycline, USP, 40 mg capsules, patients reported significant reductions in the interference of symptoms with work and social activities (P<.05). Satisfaction with doxycycline, USP, 40 mg capsules averaged 6.8 on a scale of 1 (not at all satisfied) to 9 (very satisfied) for patients using only doxycycline, USP, 40 mg capsules; mean satisfaction was 6.7 for those using it with adjunct medication. Seven patients reported 11 adverse events during the program, including lack of efficacy, joint injury with fatigue, dizziness, back pain, bloating and constipation, increased facial redness and pimples, yeast infection, sore throat, increased bruising and worsening of rosacea. Satisfaction with doxycycline, USP, 40 mg capsules for the treatment of papulopustular rosacea was apparent from patient-rated measures of treatment impact. Patients with papulopustular rosacea reported improvement in symptoms, reductions in the interference of symptoms with life's activities and satisfaction with treatment with doxycycline, USP, 40 mg capsules.


Using FLIM-FRET to measure conformational changes of transglutaminase type 2 in live cells.
Transglutaminase type 2 (TG2) is a ubiquitously expressed member of the transglutaminase family, capable of mediating a transamidation reaction between a variety of protein substrates. TG2 also has a unique role as a G-protein with GTPase activity. In response to GDP/GTP binding and increases in intracellular calcium levels, TG2 can undergo a large conformational change that reciprocally modulates the enzymatic activities of TG2. We have generated a TG2 biosensor that allows for quantitative assessment of TG2 conformational changes in live cells using Förster resonance energy transfer (FRET), as measured by fluorescence lifetime imaging microscopy (FLIM). Quantifying FRET efficiency with this biosensor provides a robust assay to quickly measure the effects of cell stress, changes in calcium levels, point mutations and chemical inhibitors on the conformation and localization of TG2 in living cells. The TG2 FRET biosensor was validated using established TG2 conformational point mutants, as well as cell stress events known to elevate intracellular calcium levels. We demonstrate in live cells that inhibitors of TG2 transamidation activity can differentially influence the conformation of the enzyme. The irreversible inhibitor of TG2, NC9, forces the enzyme into an open conformation, whereas the reversible inhibitor CP4d traps TG2 in the closed conformation. Thus, this biosensor provides new mechanistic insights into the action of two TG2 inhibitors and defines two new classes based on ability to alter TG2 conformation in addition to inhibiting transamidation activity. Future applications of this biosensor could be to discover small molecules that specifically alter TG2 conformation to affect GDP/GTP or calcium binding.


Arthritis of leprosy.
An inflammatory polyarthritis has been previously described in leprosy but accounts of the clinical characteristics have varied. All patients with joint symptoms admitted to a leprosy centre over 5 months were examined by a rheumatologist. Of 48 acute admissions, 20 (42%) had a symmetrical polyarthritis affecting the wrists and fingers. This was strikingly similar to RA in appearance. The arthritis occurred exclusively in patients with reactions mainly during anti-mycobacterial treatment. The clinical features of the joint disorder were identical in Type I (associated with alterations in immune status) and Type II (erythema nodosum leprosum) reactions. Synovial tissue from one patient revealed no mycobacteria. Four of five hand X-rays suggested small erosions of the finger joints.


Problems faced by patients and health service utilization experiences of gastrointestinal patients during lockdown due to COVID-19 pandemic.
The COVID-19 pandemic undermined the health service delivery and utilization of essential health care services globally. The current study therefore aimed to explore the health-seeking behaviors and challenges faced by patients for the management of gastrointestinal diseases. A cross-sectional study was conducted at the outpatient department of Gastroenterology, Liaquat National Hospital, Karachi from March 2020 to July 2020 during the COVID-19 lockdown phase to explore patient experiences. Data was collected using a survey questionnaire. All patients of either gender were included after informed consent. Statistical analysis of the data was conducted using SPSS 21.0. A total of 184 patients were included who visited the hospital to seek medical services during the COVID-19 lockdown phase. The mean age of the population was 42.7 years (±16.13). Of these, n=94 (51.1%) were males All patients had gastrointestinal issues with different comorbid conditions. One forty-seven n=147 (79.9%) presented with active complaints whereas, n=37 (20.1%) patients visited the hospital for their follow-up checkup. Out of 184 patients, n=33 (17.9%) patients reported of having fear of visiting hospital due to COVID-19 outbreak. A statistically significant difference p<0.001 was noted between the history of comorbidities and patient delaying a visit to the healthcare due to the fear of COVID-19. Additionally, 61 (73.5%) patients with co-morbidity faced difficulty in finding public transport (p=0.01). Nevertheless, n=171 (93.0%) patients expressed satisfaction with the services provided by the hospital during the lockdown phase. Patients with gastrointestinal conditions were largely affected by lockdown largely due to fear of contacting COVID-19 disease and inaccessibility to the public transportation. Widely available telemedicine service might overcome these shortcomings and ensure continuity of quality care.


Coordination polymers utilizing N-oxide functionalized host ligands.
Pyridyl functionalized host molecules are oxidized to their N-oxide analogues and form a series of coordination polymers and discrete complexes with transition metal cations. Complex {[Ag(3)(NMP)(6)(L1)(2)]·3(ClO(4))}(∞) where L1 = tris(isonicotinoyl-N-oxide)cyclotriguaiacylene, NMP = N-methylpyrrolidone, is a three-dimensional (3-D) 3,6-connected coordination polymer of pyrite-like (pyr) topology and features ligand unsupported argentophilic interactions, while two-dimensional (2-D) 3,6-connected coordination polymers with the rarely reported kagome dual (kgd) topology are found for [M(L1)(2)](2+) where M = Zn, Cd, Cu. Ligand L2 = tris(nicotinoyl-N-oxide)cyclotriguaiacylene forms a 2-D coordination polymer with 4(4) (sql) grid topology in complexes {[M(L2)(2)(DMF)(2)]·2ClO(4)·8(DMF)}(∞) M = Cd or Cu, DMF = N,N'-dimethylformamide, and a double-linked chain structure in {[Co(L2)(2)(DMF)(2)]·2NO(3)·4(DMF)·H(2)O}(∞), and both types of structure feature hand-shake self-inclusion motifs either within or between the polymers. 2-D coordination networks with 6(3) (hcb) topologies are found in complexes {[M(L3)(NO(3))(2)]·2(DMF)}(∞) (M = Cd, Zn) and {[Cu(5)(L3)(2)Cl(10)(NMP)(4)]}(∞) where L3 = tris(2-pyridylmethyl)cyclotriguaiacylene, while [Ag(2)(L3)(2)(NMP)(4)]·2(BF(4))·2(NMP) has a discrete dimeric structure which again shows hand-shake host-guest interactions supported by π-π stacking.


Accelerating phase contrast MR angiography by simplified skipped phase encoding and edge deghosting with array coil enhancement.
The aim of this work is to investigate the feasibility of accelerating phase contrast magnetic resonance angiography (PC-MRA) by the fast imaging method of simplified skipped phase encoding and edge deghosting with array coil enhancement (S-SPEED-ACE). The parallel imaging method of skipped phase encoding and edge deghosting with array coil enhancement (SPEED-ACE) is simplified for imaging sparse objects like phase contrast MRA. This approach is termed S-SPEED-ACE in which k-space is sparsely sampled with skipped phase encoding at every Nth step using multiple receiver coils simultaneously. The sampled data are then Fourier transformed into a set of ghosted images, each with N-fold aliasing ghosts. Given signal sparseness of MRA, the ghosted images are modeled with a single-layer structure, in which the most dominant ghost within the potentially overlapped ghosts at each pixel is selected to represent the signal of that pixel. The single-layer model is analogous to that used in maximum-intensity-projection (MIP) that selects only the brightest signal even when there are overlapped vessels. With an algorithm based on a least-square-error solution, a deghosted image is obtained, along with a residual map for quality control. In this way, S-SPEED-ACE partially samples k-space using multiple receiver coils in parallel, and yields a deghosted image with an acceleration factor of N. Without full central k-space sampling and differential filtering, S-SPEED-ACE achieves further scan time reduction with a more straightforward reconstruction. In this work, S-SPEED-ACE is demonstrated to accelerate a computer simulated PC-MRA and a real human 3D PC-MRA, which was acquired using a clinical 1.5 T scanner on a healthy volunteer. Images are reconstructed by S-SPEED-ACE to achieve an undersampling factor of up to 8.3 with four receiver coils. The reconstructed images generally have comparable quality as that of the reference images reconstructed from full k-space data. Maximum-intensity-projection images generated from the reconstructed images also demonstrated to be consistent as those from the reference images. By taking advantage of signal sparsity naturally existing in the data, SPEED-ACE was simplified and its efficiency was improved. The feasibility of the proposed S-SPEED-ACE is demonstrated in this work with simulated sampling of an actual 3D head PC-MRA scan.


Medical consult: aHUS, TTP? How to distinguish and what to do.
Immune thrombotic thrombocytopenic purpura (iTTP) caused by an autoantibody-mediated deficiency of ADAMTS13 and atypical hemolytic syndrome (aHUS) caused by alternative complement dysregulation are the most common primary thrombotic microangiopathies (TMAs). The evaluation of a patient with TMA is a medical emergency since it is critical to quickly distinguish iTTP and aHUS from other causes of TMA. Untreated iTTP is rapidly fatal, and delays in initiating complement inhibition in aHUS increase the risk of irreversible renal failure. An ADAMTS13 activity level of less than 10% is diagnostic of iTTP in the appropriate clinical setting. In settings where rapid-turnaround ADAMTS13 testing is not available, clinical features and clinical prediction tools are useful to identify patients who should receive emergent plasma exchange. We present an evidence-based approach to the initial (first 24 hours) diagnosis and management of iTTP and review the clinical and laboratory features that can be used to identify patients with aHUS who will benefit from early C5 blockade. We also discuss the potential use of complement blockade to improve outcomes in selected patients with secondary TMA.


Testing the validity of three submaximal ergometer tests for estimating maximal aerobic capacity in children.
Aerobic capacity in children has been linked to health-related outcomes, but the validity of existing assessment methods is largely unknown. This study evaluated the validity of the Åstrand-Rhyming, Woynarowska and Olgun Binyildiz methods. Aerobic capacity was estimated from the heart rate response to submaximal ergometer cycling in 62 children aged 11-12 years. Direct measurement of peak oxygen consumption during a graded maximal treadmill test was used as the criterion method. We found low mean bias for age-adjusted Åstrand-Rhyming data and Woynarowska data, (-14 and 23 mL/min, respectively), low correlation to criterion values (0.81 and 0.74, respectively) and high standard error of estimate (SEE) (340 and 395 mL/min). The Olgun Binyildiz method gave high correlation (0.87) and low SEE (298 mL/min), but large bias (-660 mL/min). All methods underestimated capacity in well-trained children. The Olgun Binyildiz method is recommended for following an individual over time, due to its low random error. But for comparing individual data with those obtained using direct measurements, the Woynarowska or Åstrand-Rhyming method may be the first choice. However, they all underestimated aerobic capacity in well-trained children.


Peer review of grant applications: criteria used and qualitative study of reviewer practices.
Peer review of grant applications has been criticized as lacking reliability. Studies showing poor agreement among reviewers supported this possibility but usually focused on reviewers' scores and failed to investigate reasons for disagreement. Here, our goal was to determine how reviewers rate applications, by investigating reviewer practices and grant assessment criteria. We first collected and analyzed a convenience sample of French and international calls for proposals and assessment guidelines, from which we created an overall typology of assessment criteria comprising nine domains relevance to the call for proposals, usefulness, originality, innovativeness, methodology, feasibility, funding, ethical aspects, and writing of the grant application. We then performed a qualitative study of reviewer practices, particularly regarding the use of assessment criteria, among reviewers of the French Academic Hospital Research Grant Agencies (Programmes Hospitaliers de Recherche Clinique, PHRCs). Semi-structured interviews and observation sessions were conducted. Both the time spent assessing each grant application and the assessment methods varied across reviewers. The assessment criteria recommended by the PHRCs were listed by all reviewers as frequently evaluated and useful. However, use of the PHRC criteria was subjective and varied across reviewers. Some reviewers gave the same weight to each assessment criterion, whereas others considered originality to be the most important criterion (12/34), followed by methodology (10/34) and feasibility (4/34). Conceivably, this variability might adversely affect the reliability of the review process, and studies evaluating this hypothesis would be of interest. Variability across reviewers may result in mistrust among grant applicants about the review process. Consequently, ensuring transparency is of the utmost importance. Consistency in the review process could also be improved by providing common definitions for each assessment criterion and uniform requirements for grant application submissions. Further research is needed to assess the feasibility and acceptability of these measures.


Unusual c-fos induction upon chromaffin PC12 differentiation by sodium butyrate: loss of fos autoregulatory function.
Induction of PC12 pheochromocytoma cells neuronal differentiation upon treatment with nerve growth factor (NGF) is accompanied by a coupled stimulation of c-fos and c-jun oncogene transcription. We found that induction of c-fos and c-jun proto-oncogene mRNAs levels following the endocrine differentiation of PC12 cells by sodium butyrate is uncoupled. While c-fos mRNA level increased within minutes, the content of c-jun mRNA was significantly elevated only 24 hours after treatment. Continuous presence of sodium butyrate for 72 hours resulted in stable high levels of c-fos and c-jun mRNAs. Gene transcription of the other members of the jun family, jun B and jun D, was not significantly modified at any induction time. The early accumulation of c-fos mRNA was accompanied by increased levels of c-Fos protein. While the NGF-induced c-Fos protein migrates with an apparent homogeneous molecular weight of 62 kDa, the sodium butyrate-stimulated Fos protein is of heterogeneous lower molecular weight. The different gel mobility of the Fos immunoreactive bands induced by sodium butyrate and the sustained Fos mRNA levels after induction suggested that the sodium butyrate-induced c-Fos protein could be non-functional in the autoregulation of the c-fos gene. Gel shift analysis showed unimpaired capacity of the butyrate-induced c-Fos protein to participate in the formation of transcriptional complexes with the Jun/AP-1 protein. However, transfection experiments indicate that the sodium butyrate-induced c-Fos protein is not able to negatively trans-regulate the c-fos promoter.


Hypermethylated FAM5C and MYLK in serum as diagnosis and pre-warning markers for gastric cancer.
Most cases of gastric cancer (GC) are not diagnosed at early stage which can be curable, so it is necessary to identify effective biomarkers for its diagnosis and pre-warning. We have used methylated DNA immunoprecipitation (MeDIP) to identify genes that are frequently methylated in gastric cancer cell lines. Promoter regions hypermethylation of candidate genes were tested by methylation-specific polymerase chain reaction (MSP) in serum samples, including GC (n=58), gastric precancerous lesions (GPL, n=46), and normal controls (NC, n=30). Eighty two hypermethylated genes were acquired by array analysis and 5 genes (BCAS4, CHRM2, FAM5C, PRAC and MYLK) were selected as the candidate genes. Three genes (CHRM2, FAM5C and MYLK) were further confirmed to show methylation rates increased with progression from NC to GPL, then to GC. There was obvious decrease in detection of FAM5C and MYLK hypermethylation, but not CHRM2, from preoperative to postoperative evaluation (P< 0.001). Combined detection of FAM5C and MYLK hypermethylation had a higher sensitivity in GC diagnosis (77.6%,45/58) and pre-warning (30.4%,14/46) than one single gene detection and also had a high specificity of 90%. The combined hypermethylated status of FAM5C and MYLK correlated with tumor size (P<0.001), tumor invasion depth (P=0.001) and tumor-node-metastasis (TNM) stage (P=0.003). Hypermethylated FAM5C and MYLK can be used as potential biomarkers for diagnosis and pre-warning of GC.


Hemagglutinin receptor binding avidity drives influenza A virus antigenic drift.
Rapid antigenic evolution in the influenza A virus hemagglutinin precludes effective vaccination with existing vaccines. To understand this phenomenon, we passaged virus in mice immunized with influenza vaccine. Neutralizing antibodies selected mutants with single-amino acid hemagglutinin substitutions that increased virus binding to cell surface glycan receptors. Passaging these high-avidity binding mutants in naïve mice, but not immune mice, selected for additional hemagglutinin substitutions that decreased cellular receptor binding avidity. Analyzing a panel of monoclonal antibody hemagglutinin escape mutants revealed a positive correlation between receptor binding avidity and escape from polyclonal antibodies. We propose that in response to variation in neutralizing antibody pressure between individuals, influenza A virus evolves by adjusting receptor binding avidity via amino acid substitutions throughout the hemagglutinin globular domain, many of which simultaneously alter antigenicity.


Nonlinear dynamic and pattern bifurcations in a model for spatial patterns in young mussel beds.
Young mussel beds on soft sediments can display large-scale regular spatial patterns. This phenomenon can be explained relatively simply by a reaction-diffusion-advection (RDA) model of the interaction between algae and mussel, which includes the diffusive spread of mussel and the advection of algae. We present a detailed analysis of pattern formation in this RDA model. We derived the conditions for differential-flow instability that cause the formation of spatial patterns, and then systematically investigated how these patterns depend on model parameters. We also present a detailed study of the patterned solutions in the full nonlinear model, using numerical bifurcation analysis of the ordinary differential equations, which were obtained from the RDA model. We show that spatial patterns occur for a wide range of algal concentrations, even when algal concentration is much lower than the prediction by linear analysis in the RDA model. That is to say, spatial patterns result from the interaction of nonlinear terms. Moreover, patterns with different wavelength, amplitude and movement speed may coexist. The results obtained are consistent with the previous observation that self-organization allows mussels to persist with algal concentrations that would not permit survival of mussels in a homogeneous bed.


Sentinel node detection in cervical cancer with (99m)Tc-phytate.
The aim of this study was to investigate the feasibility of sentinel lymph node (SLN) identification using radioisotopic lymphatic mapping with technetium-99 m-labeled phytate in patients undergoing radical hysterectomy with pelvic lymphadenectomy for treatment of early cervical cancer. Between July 2001 and February 2003, 56 patients with cervical cancer FIGO stage I (n = 53) or stage II (n = 3) underwent sentinel lymph node detection with preoperative lymphoscintigraphy ((99m)Tc-labeled phytate injected into the uterine cervix, at 3, 6, 9, and 12 o'clock, at a dose of 55-74 MBq in a volume of 0.8 ml) and intraoperative lymphatic mapping with a handheld gamma probe. Radical hysterectomy was aborted in three cases because parametrial invasion was found intraoperatively and we performed only sentinel node resection. The remaining 53 patients underwent radical hysterectomy with complete pelvic lymphadenectomy. Sentinel nodes were detected using a handheld gamma-probe and removed for pathological assessment during the abdominal radical hysterectomy and pelvic lymphadenectomy. One or more sentinel nodes were detected in 52 out of 56 eligible patients (92.8%). A total of 120 SLNs were detected by lymphoscintigraphy (mean 2.27 nodes per patient) and intraoperatively by gamma probe. Forty-four percent of SLNs were found in the external iliac area, 39% in the obturator region, 8.3% in interiliac region, and 6.7% in the common iliac area. Unilateral sentinel nodes were found in thirty-one patients (59%). The remaining 21 patients (41%) had bilateral sentinel nodes. Microscopic nodal metastases were confirmed in 17 (32%) cases. In 10 of these patients, only SLNs had metastases. The 98 sentinel nodes that were negative on hematoxylin and eosin were submitted to cytokeratin immunohistochemical analysis. Five (5.1%) micrometastases were identified with this technique. The sensitivity of the sentinel node was 82.3% (CI 95% = 56.6-96.2) and the negative predictive value was 92.1% (CI 95% = 78.6-98.3). The accuracy of sentinel node in predicting the lymph node status was 94.2%. Preoperative lymphoscintigraphy and intraoperative lymphatic mapping with (99m)Tc-labeled phytate are effective in identifying sentinel nodes in patients undergoing radical hysterectomy and to select women in whom lymph node dissection can be avoided.


Beyond one-size-fits-all: Tailoring diversity approaches to the representation of social groups.
When and why do organizational diversity approaches that highlight the importance of social group differences (vs. equality) help stigmatized groups succeed? We theorize that social group members' numerical representation in an organization, compared with the majority group, influences concerns about their distinctiveness, and consequently, whether diversity approaches are effective. We combine laboratory and field methods to evaluate this theory in a professional setting, in which White women are moderately represented and Black individuals are represented in very small numbers. We expect that focusing on differences (vs. equality) will lead to greater performance and persistence among White women, yet less among Black individuals. First, we demonstrate that Black individuals report greater representation-based concerns than White women (Study 1). Next, we observe that tailoring diversity approaches to these concerns yields greater performance and persistence (Studies 2 and 3). We then manipulate social groups' perceived representation and find that highlighting differences (vs. equality) is more effective when groups' representation is moderate, but less effective when groups' representation is very low (Study 4). Finally, we content-code the diversity statements of 151 major U.S. law firms and find that firms that emphasize differences have lower attrition rates among White women, whereas firms that emphasize equality have lower attrition rates among racial minorities (Study 5). (PsycINFO Database Record


Synchronization in a noise-driven developing neural network.
We use computer simulations to investigate the structural and dynamical properties of a developing neural network whose activity is driven by noise. Structurally, the constructed neural networks in our simulations exhibit the small-world properties that have been observed in several neural networks. The dynamical change of neuronal membrane potential is described by the Hodgkin-Huxley model, and two types of learning rules, including spike-timing-dependent plasticity (STDP) and inverse STDP, are considered to restructure the synaptic strength between neurons. Clustered synchronized firing (SF) of the network is observed when the network connectivity (number of connections/maximal connections) is about 0.75, in which the firing rate of neurons is only half of the network frequency. At the connectivity of 0.86, all neurons fire synchronously at the network frequency. The network SF frequency increases logarithmically with the culturing time of a growing network and decreases exponentially with the delay time in signal transmission. These conclusions are consistent with experimental observations. The phase diagrams of SF in a developing network are investigated for both learning rules.


Dynamic intra-articular space variation in clicking TMJs.
During mandibular movement, the geometric relationships of the articular surfaces in the temporomandibular joint (TMJ) change, so that the disc undergoes different stress concentrations with respect to time and position. In this study, we compared the intra-articular space variations of 13 clicking and 15 asymptomatic TMJs for jaw opening/closing. Magnetic resonance imaging and jaw tracking were combined to display the motion of the whole condyle within the fossa. In clicking TMJs, the mediolateral spread s of the stress-field trajectories was 2.4 +/- 1.0 mm (s(max) = 4.9 +/- 2.1 mm) with an aspect ratio a/h of 2.5 +/- 1.6, both significantly greater than in controls (p < 0.05). The stress-field trajectories of the controls coincided during opening/closing (s = 0.9 +/- 0.2 mm, s(max) = 1.8 +/- 0.8 mm, a/h = 1.6 +/- 0.3). Clicking TMJs showed much less coincident stress-field paths and much "flatter" stress-fields than controls during jaw opening/closing.


Metal toxin threat in wildland fires determined by geology and fire severity.
Accentuated by climate change, catastrophic wildfires are a growing, distributed global public health risk from inhalation of smoke and dust. Underrecognized, however, are the health threats arising from fire-altered toxic metals natural to soils and plants. Here, we demonstrate that high temperatures during California wildfires catalyzed widespread transformation of chromium to its carcinogenic form in soil and ash, as hexavalent chromium, particularly in areas with metal-rich geologies (e.g., serpentinite). In wildfire ash, we observed dangerous levels (327-13,100 µg kg-1) of reactive hexavalent chromium in wind-dispersible particulates. Relatively dry post-fire weather contributed to the persistence of elevated hexavalent chromium in surficial soil layers for up to ten months post-fire. The geographic distribution of metal-rich soils and fire incidents illustrate the broad global threat of wildfire smoke- and dust-born metals to populations. Our findings provide new insights into why wildfire smoke exposure appears to be more hazardous to humans than pollution from other sources.


Studying the Association between Sudden Hearing Loss and DNA N-Methyltransferase 1 (DNMT1) Genetic Polymorphism.
The aim of the present study was to investigate whether there was any relationship between some DNA N-methyltransferase 1 (DNMT1) polymorphisms and susceptibility to idiopathic sudden sensorineural hearing loss (ISSHL) in ISSHL patients. We investigated 90 patients diagnosed with ISSHL and a control group composed of 75 age- and gender-matched healthy individuals. DNA was extracted from the blood samples by phenol-chloroform method. Polymerase chain reaction and restriction fragment length polymorphism methods were used for the genotyping analysis of 4 regions of DNMT1. For rs2228612 single nucleotide polymorphism (SNP), the frequency of AA, AG, and GG genotypes were 81.4%, 9.3%, and 9.3% in controls and 82.2%, 16.7%, and 1.1% in patients, respectively. We observed a significant decrease in the frequency of GG genotype in patients with ISSHL when compared with controls (p=0.027). The frequency of GG, AG, and AA genotypes for rs2228611 SNP were 20.7%, 49.3%, and 20% in controls and 20%, 47.8%, and 32.2% in patients, respectively. There was a significantly increased frequency of the AA genotype of this SNP in the DNMT1 gene, and we found that individuals with the AA genotype had 2.47 times the risk for ISSHL development than individuals with the GG genotype (p=0.41). The GAA haplotype may constitute 2.66 times the risk for ISSHL disease (OR=2.66, 95% confidence interval: 0.28-25.03). This study's results showed that the AA genotype in rs2228611 polymorphism was a risk factor in ISSHL patients and the GG genotype could be a protective factor in rs2228612 polymorphism.


Responses of blood lipids to aerobic, resistance, and combined aerobic with resistance exercise training: a systematic review of current evidence.
This review considers the effectiveness of aerobic exercise training with different intensities (moderate and high) as well as the type of exercise (aerobic, resistance, and combined aerobic with resistance) in altering the blood lipids. We reviewed various trials via a systematic search of PubMed, published reviews, and references from original articles. We selected studies that involved aerobic and/or resistance and/or combined exercise training in healthy adults over a limit of 12 weeks and had examined the response of training to one or more of the following: triglycerides, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol. We selected a total of 84 studies, 58 were randomized controlled trials. Comparisons between intensities of aerobic exercise programs resulted in favorable effects only for high intensity. The most frequently observed alteration was an increase in the high-density lipoprotein cholesterol, whereas reductions in triglycerides, total cholesterol, and low-density lipoprotein cholesterol appeared less often. Moreover, the evidence of the positive effect of resistance exercise marks out a trend mainly for the low-density lipoprotein cholesterol levels, whereas for combined exercise, results extracted from a short list of published studies show improvements in values of both the high-density lipoprotein cholesterol and the low-density lipoprotein cholesterol. High-intensity aerobic training results in improvement in high-density lipoprotein cholesterol. For resistance and combined exercise, the results are inconsistent. The heterogeneity between the types of exercise did not allow reliable comparisons.


Removal of endocrine-disrupting compounds from water using macroporous molecularly imprinted cryogels in a moving-bed reactor.
Highly efficient removal of endocrine-disrupting compounds (EDCs) such as 17beta-estradiol (E2), 4-nonylphenol (NP) and atrazine from water was achieved using a novel macroporous adsorption medium. The medium consisted of a macroporous poly(vinyl alcohol) (PVA) cryogel with molecularly imprinted polymer (MIP) particles embedded in it. The MIP was prepared using E2, NP and atrazine as templates. The macroporous composite molecularly imprinted cryogels were formed inside the open-ended protective shells, known as Kaldnes carriers. These adsorbents (defined as Macroporous Gel Particles, MGPs) were evaluated on the removal of E2, NP and atrazine from water using different column configurations, namely column filled with the MGPs (packed-bed column) and in moving-bed reactors (defined here as moving-bed MGPs reactor). Complete binding (> 99%) of E2 from a spiked aqueous solution (1 mg/L) was achieved using E2-MIP/MGPs in a moving-bed MGPs reactor at the retention time in the reactor of 4 min, while only 77% was bound to the nonimprinted medium (NIP/MGPs). Similar results were also obtained for the adsorption medium imprinted with atrazine. All contaminants studied (E2, atrazine and NP) were effectively removed from water at low (environmentally relevant) concentrations by the respective adsorption medium.


A systematic review of publications assessing reliability and validity of the Behavioral Risk Factor Surveillance System (BRFSS), 2004-2011.
In recent years response rates on telephone surveys have been declining. Rates for the behavioral risk factor surveillance system (BRFSS) have also declined, prompting the use of new methods of weighting and the inclusion of cell phone sampling frames. A number of scholars and researchers have conducted studies of the reliability and validity of the BRFSS estimates in the context of these changes. As the BRFSS makes changes in its methods of sampling and weighting, a review of reliability and validity studies of the BRFSS is needed. In order to assess the reliability and validity of prevalence estimates taken from the BRFSS, scholarship published from 2004-2011 dealing with tests of reliability and validity of BRFSS measures was compiled and presented by topics of health risk behavior. Assessments of the quality of each publication were undertaken using a categorical rubric. Higher rankings were achieved by authors who conducted reliability tests using repeated test/retest measures, or who conducted tests using multiple samples. A similar rubric was used to rank validity assessments. Validity tests which compared the BRFSS to physical measures were ranked higher than those comparing the BRFSS to other self-reported data. Literature which undertook more sophisticated statistical comparisons was also ranked higher. Overall findings indicated that BRFSS prevalence rates were comparable to other national surveys which rely on self-reports, although specific differences are noted for some categories of response. BRFSS prevalence rates were less similar to surveys which utilize physical measures in addition to self-reported data. There is very little research on reliability and validity for some health topics, but a great deal of information supporting the validity of the BRFSS data for others. Limitations of the examination of the BRFSS were due to question differences among surveys used as comparisons, as well as mode of data collection differences. As the BRFSS moves to incorporating cell phone data and changing weighting methods, a review of reliability and validity research indicated that past BRFSS landline only data were reliable and valid as measured against other surveys. New analyses and comparisons of BRFSS data which include the new methodologies and cell phone data will be needed to ascertain the impact of these changes on estimates in the future.


Oxaliplatin resistance is enhanced by saracatinib via upregulation Wnt-ABCG1 signaling in hepatocellular carcinoma.
Chemo-resistance in hepatocellular carcinoma (HCC) is a major problem, and acquired drug resistance prevents cancer therapies from achieving complete responses. Molecular targeting therapy presents an opportunity to impede tumor through combination or sequential therapy, while the accurate effect is vague. The efficacy of combinations between oxaliplatin and anti-cancer molecular targeting drugs was screened. Strangely, the combined chemotherapy with oxaliplatin and saracatinib induced significantly antagonistic effects. Then the antitumor effects of combined treatment with saracatinib and oxaliplatin were confirmed in wide type HCC as well as in saracatinib- and oxaliplatin-resistant HCC. RNA sequencing was used to explore the resistance mechanism, and the roles of ATP-binding cassette transporter G1 (ABCG1) and Wnt signaling in oxaliplatin resistance were confirmed. Chemotherapy with oxaliplatin and saracatinib individually induced strong anti-HCC effects, while combined or sequential treatment of HCC cells with these two drugs exhibited reduced efficacy compared to treatment with the single drugs. And it was saracatinib treatment caused oxaliplatin resistance. RNA sequencing revealed 458 genes that were altered by treatment with saracatinib and oxaliplatin. Of these, the gene encoding ABCG1 and Wnt-associated genes were significantly upregulated. Upregulation of ABCG1 and oxaliplatin resistance were associated with activation of Wnt signaling. Interference with ABCG1 expression or inhibition of Wnt signaling resulted in reversal of the saracatinib-induced oxaliplatin resistance in HCC. These studies demonstrated that combined or sequential chemotherapy with oxaliplatin and saracatinib reduced antitumor efficacy, and this antagonism was attributed to the activation of Wnt signaling and upregulation of ABCG1 by saracatinib.


Interventions for female drug-using offenders.
This is an updated version of a Cochrane review first published in Issue 3, 2006 (Perry 2006). The review represents one in a family of four reviews focusing on the effectiveness of interventions in reducing drug use and criminal activity for offenders. This specific review considers interventions for female drug-using offenders. To assess the effectiveness of interventions for female drug-using offenders in reducing criminal activity, or drug use, or both. We searched 14 electronic bibliographic databases up to May 2014 and five additional Website resources (between 2004 and November 2011). We contacted experts in the field for further information. We included randomised controlled trials (RCTs) designed to reduce, eliminate or prevent relapse of drug use or criminal activity in female drug-using offenders. We also reported data on the cost and cost-effectiveness of interventions. We used standard methodological procedures expected by The Cochrane Collaboration. Nine trials with 1792 participants met the inclusion criteria. Trial quality and risks of bias varied across each study. We rated the majority of studies as being at 'unclear' risk of bias due to a lack of descriptive information. We divided the studies into different categories for the purpose of meta-analyses: for any psychosocial treatments in comparison to treatment as usual we found low quality evidence that there were no significant differences in arrest rates, (two studies; 489 participants; risk ratio (RR) 0.82, 95% confidence interval (CI) 0.45 to 1.52) or drug use (one study; 77 participants; RR 0.65, 95% CI 0.20 to 2.12), but we found moderate quality evidence that there was a significant reduction in reincarceration, (three studies; 630 participants; RR 0.46, 95% CI 0.34 to 0.64). Pharmacological intervention using buprenorphine in comparison to a placebo did not significantly reduce self reported drug use (one study; 36 participants; RR 0.58, 95% CI 0.25 to 1.35). No cost or cost-effectiveness evidence was reported in the studies. Three of the nine trials show a positive trend towards the use of any psychosocial treatment in comparison to treatment as usual showing an overall significant reduction in subsequent reincarceration, but not arrest rates or drug use. Pharmacological interventions in comparison to a placebo did not significantly reduce drug use and did not measure criminal activity. Four different treatment comparisons showed varying results and were not combined due to differences in the intervention and comparison groups. The studies overall showed a high degree of heterogeneity for types of comparisons and outcome measures assessed, which limited the possibility to pool the data. Descriptions of treatment modalities are required to identify the important elements for treatment success in drug-using female offenders. More trials are required to increase the precision of confidence with which we can draw conclusions about the effectiveness of treatments for female drug-using offenders.


Calyculin: Nature's way of making the sponge-derived cytotoxin.
Covering: up to 2015.Calyculin A is a major cytotoxic compound isolated from the Japanese marine sponge Discodermia calyx. Its potent cytotoxicity is attributable to the specific inhibition of protein phosphatases 1 and 2A, as in the case of okadaic acid and the microcystins. Its chemical structure is well-designed not only for enzyme inhibition but also for higher membrane permeability in order to impart its potent cytotoxicity. The biosynthetic gene cluster of this densely functionalized polyketide and nonribosomal peptide hybrid molecule was recently identified from the sponge-microbe association. The producer organism and the dynamic bioconversion process were also revealed. In this highlight, we focus on the recent studies addressing nature's design and biogenesis of the sponge-derived cytotoxin, calyculin A.


A Novel Zinc Chelator, 1H10, Ameliorates Experimental Autoimmune Encephalomyelitis by Modulating Zinc Toxicity and AMPK Activation.
Previous studies in our lab revealed that chemical zinc chelation or zinc transporter 3 (ZnT3) gene deletion suppresses the clinical features and neuropathological changes associated with experimental autoimmune encephalomyelitis (EAE). In addition, although protective functions are well documented for AMP-activated protein kinase (AMPK), paradoxically, disease-promoting effects have also been demonstrated for this enzyme. Recent studies have demonstrated that AMPK contributes to zinc-induced neurotoxicity and that 1H10, an inhibitor of AMPK, reduces zinc-induced neuronal death and protects against oxidative stress, excitotoxicity, and apoptosis. Here, we sought to evaluate the therapeutic efficacy of 1H10 against myelin oligodendrocyte glycoprotein 35-55-induced EAE. 1H10 (5 μg/kg) was intraperitoneally injected once per day for the entire experimental course. Histological evaluation was performed three weeks after the initial immunization. We found that 1H10 profoundly reduced the severity of the induced EAE and that there was a remarkable suppression of demyelination, microglial activation, and immune cell infiltration. 1H10 also remarkably inhibited EAE-associated blood-brain barrier (BBB) disruption, MMP-9 activation, and aberrant synaptic zinc patch formation. Furthermore, the present study showed that long-term treatment with 1H10 also reduced the clinical course of EAE. Therefore, the present study suggests that zinc chelation and AMPK inhibition with 1H10 may have great therapeutic potential for the treatment of multiple sclerosis.


Linear and dendrimeric antiviral peptides: design, chemical synthesis and activity against human respiratory syncytial virus.
Respiratory syncytial virus (RSV) is one of the most common viral pathogens. It is especially dangerous for newborns and young children. In some cases it could lead to severe bronchiolitis, pneumonia with hospitalization or even a lethal outcome. Despite decades of investigation of RSV biology, effective and safe therapeutics are still under development. Certain natural peptides have been found to exhibit antiviral activity against respiratory viruses, but their implementation is limited by low stability in biological media. One of the current approaches to enhance the peptide therapeutic opportunities is chemical synthesis of peptide dendrimers with hyperbranched structures. Taking into account the recent data of bioactive cationic and helical regions of natural peptides and the structure features of nucleolin identified as an RSV cellular receptor, the main goal of this study was to design relatively short linear and dendrimeric cationic peptides and to test their antiviral activity against RSV. As a result 3 linear cationic peptides and 4 peptide dendrimers were synthesized and compared with known LL-37 (cathelicidin family) and anti-F0 monoclonal antibodies in terms of cytotoxicity and antiviral activity. Their affinity to the supposed molecular target - nucleolin (C23) - was estimated in silico by molecular docking analysis. Four synthesized peptides demonstrated a cytotoxic effect, two of them were even more cytotoxic than LL-37, which could be explained by a combination of a high amount of positive charge and amphipathicity. Contrariwise, non-hydrophobic dendrimer peptides did not exhibit cytotoxicity in mammalian cells in the studied concentration range. Two of the seven synthesized peptides, LTP (dendrimer) and SA-35 (linear), used in this study had a stronger antiviral effect than natural peptide LL-37, and three others showed slightly lower activity than anti-F0 monoclonal antibodies. The data obtained in this study suggest that evenly distributed positive charge, and low or medium amphipathicity play a key role in the antiviral activity of the studied peptides. Moreover, the calculated free energy values of the peptide/nucleolin complex for the most active peptides supported the idea that the peptide ability of nucleolin interaction promotes the anti-RSV properties of the molecules.


Endurance training attenuates the oxidative stress due to acute exhaustive exercise in rat liver.
The aim of this study was to investigate whether an 8-week treadmill training attenuates exercise-induced oxidative stress in rat liver. Male rats were divided into untrained and trained groups. Endurance training consisted of treadmill running at a speed of 2.1 km/h, 1.5 h/day, 5 days a week for 8 weeks. To see the effects of endurance training on acute exhaustive exercise induced oxidative stress, untrained and trained rats were further devided into two groups: animals killed at rest and those killed after acute exhaustive exercise, in which the rats run at 2.1 km/h (10% uphill) until exhaustion. Acute exhaustive exercise increased malondialdehyde level in untrained but not in trained rats. It decreased the activity of glutathione peroxidase and total (enzymatic plus non-enzymatic) superoxide scavenger activity in untrained rats and catalase activity in trained rats. However, it did not affect glutathione S-transferase, glutathione reductase, superoxide dismutase and non-enzymatic superoxide radical scavenger activities in both trained and untrained rats. On the other hand, endurance training decreased glutathione peroxidase and glutathione S-transferase activities. The results suggested that endurance training attenuated exercise-induced oxidative stress in liver, probably by preventing the decreases in glutathione peroxidase and total superoxide scavenger activities during exercise.


The National NeuroAIDS Tissue Consortium brain gene array: two types of HIV-associated neurocognitive impairment.
The National NeuroAIDS Tissue Consortium (NNTC) performed a brain gene expression array to elucidate pathophysiologies of Human Immunodeficiency Virus type 1 (HIV-1)-associated neurocognitive disorders. Twenty-four human subjects in four groups were examined A) Uninfected controls; B) HIV-1 infected subjects with no substantial neurocognitive impairment (NCI); C) Infected with substantial NCI without HIV encephalitis (HIVE); D) Infected with substantial NCI and HIVE. RNA from neocortex, white matter, and neostriatum was processed with the Affymetrix® array platform. With HIVE the HIV-1 RNA load in brain tissue was three log(10) units higher than other groups and over 1,900 gene probes were regulated. Interferon response genes (IFRGs), antigen presentation, complement components and CD163 antigen were strongly upregulated. In frontal neocortex downregulated neuronal pathways strongly dominated in HIVE, including GABA receptors, glutamate signaling, synaptic potentiation, axon guidance, clathrin-mediated endocytosis and 14-3-3 protein. Expression was completely different in neuropsychologically impaired subjects without HIVE. They had low brain HIV-1 loads, weak brain immune responses, lacked neuronally expressed changes in neocortex and exhibited upregulation of endothelial cell type transcripts. HIV-1-infected subjects with normal neuropsychological test results had upregulation of neuronal transcripts involved in synaptic transmission of neostriatal circuits. Two patterns of brain gene expression suggest that more than one pathophysiological process occurs in HIV-1-associated neurocognitive impairment. Expression in HIVE suggests that lowering brain HIV-1 replication might improve NCI, whereas NCI without HIVE may not respond in kind; array results suggest that modulation of transvascular signaling is a potentially promising approach. Striking brain regional differences highlighted the likely importance of circuit level disturbances in HIV/AIDS. In subjects without impairment regulation of genes that drive neostriatal synaptic plasticity reflects adaptation. The array provides an infusion of public resources including brain samples, clinicopathological data and correlative gene expression data for further exploration (http://www.nntc.org/gene-array-project).


Weekday and weekend physical activity patterns of French and Spanish adolescents.
Adolescents' objectively assessed physical activity (PA) patterns during specific segments of the day remain unclear. In order to develop a clearer understanding this study examined country and gender differences in moderate to vigorous physical activity (MVPA) levels during specific segments of weekdays and weekend days, and explored the contribution of each segment to PA guidelines. Morpho-demographic, socio-economic and PA data were collected from a sample of 829 French and Spanish adolescents (45.0% Spanish; 55.2% females; 14.33±.73 years). Actigraph GT3X accelerometers were worn for seven days to assess adolescents' MVPA for three segments of weekdays (school-travel-time, school-time, and non-school-time), and weekend days (morning-time, afternoon-time and night-time). Data were analysed using multilevel modelling. The most active segments were non-school-time (29.2±17.5 min) and school-time (25.8±14.2 min) during weekdays, and morning-time (28.2±25.8 min) on weekend days. Except for school-time, Spanish adolescents were more significantly active than French adolescents during all segments. Significant gender differences were found in all segments. Country differences highlight the need to recognise cultural contexts that influence adolescents' PA. Common European-wide strategies may be insufficient to increase MVPA levels if cultural variability is not considered. Spanish and French PA intervention programmes should target girls and low-active boys during non-school-time and weekends.


Mosquito infection responses to developing filarial worms.
Human lymphatic filariasis is a mosquito-vectored disease caused by the nematode parasites Wuchereria bancrofti, Brugia malayi and Brugia timori. These are relatively large roundworms that can cause considerable damage in compatible mosquito vectors. In order to assess how mosquitoes respond to infection in compatible mosquito-filarial worm associations, microarray analysis was used to evaluate transcriptome changes in Aedes aegypti at various times during B. malayi development. Changes in transcript abundance in response to the different stages of B. malayi infection were diverse. At the early stages of midgut and thoracic muscle cell penetration, a greater number of genes were repressed compared to those that were induced (20 vs. 8). The non-feeding, intracellular first-stage larvae elicited few differences, with 4 transcripts showing an increased and 9 a decreased abundance relative to controls. Several cecropin transcripts increased in abundance after parasites molted to second-stage larvae. However, the greatest number of transcripts changed in abundance after larvae molted to third-stage larvae and migrated to the head and proboscis (120 induced, 38 repressed), including a large number of putative, immunity-related genes (approximately 13% of genes with predicted functions). To test whether the innate immune system of mosquitoes was capable of modulating permissiveness to the parasite, we activated the Toll and Imd pathway controlled rel family transcription factors Rel1 and Rel2 (by RNA interference knockdown of the pathway's negative regulators Cactus and Caspar) during the early stages of infection with B. malayi. The activation of either of these immune signaling pathways, or knockdown of the Toll pathway, did not affect B. malayi in Ae. aegypti. The possibility of LF parasites evading mosquito immune responses during successful development is discussed.


Phytophthora database 2.0: update and future direction.
The online community resource Phytophthora database (PD) was developed to support accurate and rapid identification of Phytophthora and to help characterize and catalog the diversity and evolutionary relationships within the genus. Since its release in 2008, the sequence database has grown to cover 1 to 12 loci for ≈2,600 isolates (representing 138 described and provisional species). Sequences of multiple mitochondrial loci were added to complement nuclear loci-based phylogenetic analyses and diagnostic tool development. Key characteristics of most newly described and provisional species have been summarized. Other additions to improve the PD functionality include: (i) geographic information system tools that enable users to visualize the geographic origins of chosen isolates on a global-scale map, (ii) a tool for comparing genetic similarity between isolates via microsatellite markers to support population genetic studies, (iii) a comprehensive review of molecular diagnostics tools and relevant references, (iv) sequence alignments used to develop polymerase chain reaction-based diagnostics tools to support their utilization and new diagnostic tool development, and (v) an online community forum for sharing and preserving experience and knowledge accumulated in the global Phytophthora community. Here we present how these improvements can support users and discuss the PD's future direction.


Pulmonary metastasectomy for hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is seldom resectable due to advanced status. Even though hepatectomy is feasible, a large proportional of patients may still develops extrahepatic recurrence. Pulmonary metastasis is the most common site of extrahepatic spread. Few articles have discussed the benefit of resection for lung metastasis after curative hepatectomy. We evaluated the general information and the result of lung resection for patients having lung metastasis after curative resection of HCC. Six patients who underwent pulmonary metastasectomy for HCC at Taipei Veterans General Hospital between August 1995 and May 2004 were enrolled in the study. All of them had received the hepatectomy for primary HCC. The demographic information of patients, the site and number of extrahepatic recurrence, the method of surgical intervention and the outcome after surgery were retrospectively reviewed. There were 4 men and 2 women with the mean age of 47.3 years. All of them were HBV carriers. Five patients had multiple pulmonary metastases removed by wedge resection. Two patients had bilateral lung metastases upon diagnosis of extrahepatic recurrence. The mean duration of follow-up after hepatic resection was 75.0 +/- 25.1 months (32 to 104 months). The mean survival after pulmonary resection was 47.2 +/- 34.3months (1 to 94 months). Four patients are still alive and free of the disease. One patient is alive but with the disease. One patient who refused further aggressive treatment after resection of lung metastasis died of the disease 40 months after lung resection, 77 months after hepatic resection. Lung resection for the pulmonary metastasis of HCC can result in a favorable long-term survival when there is no other intrahepatic or extraphepatic recurrence of HCC. For patients with multiple lung metastases in different lobes or different lungs, aggressive surgical resection is recommended if complete resection can be achieved.


Neoadjuvant chemotherapy and radiotherapy followed by surgery in selected patients with stage IIIB non-small-cell lung cancer: a multicentre phase II trial.
Stage IIIB non-small-cell lung cancer (NSCLC) is usually thought to be unresectable, and is managed with chemotherapy with or without radiotherapy. However, selected patients might benefit from surgical resection after neoadjuvant chemotherapy and radiotherapy. The aim of this multicentre, phase II trial was to assess the efficacy and toxicity of a neoadjuvant chemotherapy and radiotherapy followed by surgery in patients with technically operable stage IIIB NSCLC. Between September, 2001, and May, 2006, patients with pathologically proven and technically resectable stage IIIB NSCLC were sequentially treated with three cycles of neoadjuvant chemotherapy (cisplatin with docetaxel), immediately followed by accelerated concomitant boost radiotherapy (44 Gy in 22 fractions) and definitive surgery. The primary endpoint was event-free survival at 12 months. Efficacy analyses were done by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00030810. 46 patients were enrolled, with a median age of 60 years (range 28-70). 13 (28%) patients had N3 disease, 36 (78%) had T4 disease. All patients received chemotherapy; 35 (76%) patients received radiotherapy. The main toxicities during chemotherapy were neutropenia (25 patients [54%] at grade 3 or 4) and febrile neutropenia (nine [20%]); the main toxicity after radiotherapy was oesophagitis (ten patients [29%]; nine grade 2, one grade 3). 35 patients (76%) underwent surgery, with pneumonectomy in 17 patients. A complete (R0) resection was achieved in 27 patients. Peri-operative complications occurred in 14 patients, including two deaths (30-day mortality 5.7%). Seven patients required a second surgical intervention. Pathological mediastinal downstaging was seen in 11 of the 28 patients who had lymph-node involvement at enrolment, a complete pathological response was seen in six patients. Event-free survival at 12 months was 54% (95% CI 39-67). After a median follow-up of 58 months, the median overall survival was 29 months (95% CI 16.1-NA), with survival at 1, 3, and 5 years of 67% (95% CI 52-79), 47% (32-61), and 40% (24-55). A treatment strategy of neoadjuvant chemotherapy and radiotherapy followed by surgery is feasible in selected patients. Toxicity is considerable, but manageable. Survival compares favourably with historical results of combined treatment for less advanced stage IIIA disease. Swiss Group for Clinical Cancer Research (SAKK) and an unrestricted educational grant by Sanofi-Aventis (Switzerland).


Highly sensitive amplicon-based transcript quantification by semiconductor sequencing.
In clinical and basic research custom panels for transcript profiling are gaining importance because only project specific informative genes are interrogated. This approach reduces costs and complexity of data analysis and allows multiplexing of samples. Polymerase-chain-reaction (PCR) based TaqMan assays have high sensitivity but suffer from a limited dynamic range and sample throughput. Hence, there is a gap for a technology able to measure expression of large gene sets in multiple samples. We have adapted a commercially available mRNA quantification assay (AmpliSeq-RNA) that measures mRNA abundance based on the frequency of PCR amplicons determined by high-throughput semiconductor sequencing. This approach allows for parallel, accurate quantification of about 1000 transcripts in multiple samples covering a dynamic range of five orders of magnitude. Using samples derived from a well-characterized stem cell differentiation model, we obtained a good correlation (r = 0.78) of transcript levels measured by AmpliSeq-RNA and DNA-microarrays. A significant portion of low abundant transcripts escapes detection by microarrays due to limited sensitivity. Standard quantitative RNA sequencing of the same samples confirms expression of low abundant genes with an overall correlation coefficient of r = 0.87. Based on digital AmpliSeq-RNA imaging we show switches of signaling cascades at four time points during differentiation of stem cells into cardiomyocytes. The AmpliSeq-RNA technology adapted to high-throughput semiconductor sequencing allows robust transcript quantification based on amplicon frequency. Multiplexing of at least 900 parallel PCR reactions is feasible because sequencing-based quantification eliminates artefacts coming from off-target amplification. Using this approach, RNA quantification and detection of genetic variations can be performed in the same experiment.


Affinity proteomics for systematic protein profiling of chromosome 21 gene products in human tissues.
Here we show that an affinity proteomics strategy using affinity-purified antibodies raised against recombinant human protein fragments can be used for chromosome-wide protein profiling. The approach is based on affinity reagents raised toward bioinformatics-designed protein epitope signature tags corresponding to unique regions of individual gene loci. The genes of human chromosome 21 identified by the genome efforts were investigated, and the success rates for de novo cloning, protein production, and antibody generation were 85, 76, and 56%, respectively. Using human tissue arrays, a systematic profiling of protein expression and subcellular localization was undertaken for the putative gene products. The results suggest that this affinity proteomics strategy can be used to produce a proteome atlas, describing distribution and expression of proteins in normal tissues as well as in common cancers and other forms of diseased tissues.


Which Should Be the Preferred Technique During Laparoscopic Ovarian Cystectomy.
The aim of the present study was to determine the long-term effects of different laparoscopic hemostatic techniques on ovarian reserve after ovarian cystectomy. Ninety patients with unilateral ovarian cysts were recruited and randomly distributed into 2 groups. Laparoscopic stripping cystectomy was performed in all patients. Afterward, cystectomy hemostasis was achieved via hemostatic suture or bipolar electrocoagulation. Serum levels of anti-Müllerian hormone (AMH) were determined preoperatively and postoperatively at 1, 3, and 12 months, and patients were evaluated for residual ovarian volume, antral follicle count, and pregnancy. The statistical difference was determined between the 2 groups in terms of AMH levels at 3 months (hemostatic suture group = 3.17 ± 3.40 vs bipolar electrocoagulation group = 2.38 ± 2.57, P = .006) and 12 months (hemostatic suture group = 3.71 ± 3.09 vs bipolar electrocoagulation group = 2.78 ± 2.85, P = .005). In addition, in the hemostatic suture group, there was no statistically significant difference between preoperative and postoperative AMH levels ( P = .165) and between the postoperative antral follicle count ( P = .779) and the residual ovarian volume ( P = .248), whereas in the bipolar electrocoagulation group, postoperative AMH levels were lower than preoperative levels ( P = .028) and postoperative residual ovarian volumes at 3 and 12 months were lower than those at 1 month ( P = .001). Nonetheless, pregnancy rates were not significantly different ( P = .546). Bipolar electrocoagulation is more destructive compared with hemostatic suture. However, the ovarian reserve does not decrease further during the follow-up period.


Mental Rotation of Digitally-Rendered Haptic Objects.
Sensory substitution is an effective means to rehabilitate many visual functions after visual impairment or blindness. Tactile information, for example, is particularly useful for functions such as reading, mental rotation, shape recognition, or exploration of space. Extant haptic technologies typically rely on real physical objects or pneumatically driven renderings and thus provide a limited library of stimuli to users. New developments in digital haptic technologies now make it possible to actively simulate an unprecedented range of tactile sensations. We provide a proof-of-concept for a new type of technology (hereafter haptic tablet) that renders haptic feedback by modulating the friction of a flat screen through ultrasonic vibrations of varying shapes to create the sensation of texture when the screen is actively explored. We reasoned that participants should be able to create mental representations of letters presented in normal and mirror-reversed haptic form without the use of any visual information and to manipulate such representations in a mental rotation task. Healthy sighted, blindfolded volunteers were trained to discriminate between two letters (either L and P, or F and G; counterbalanced across participants) on a haptic tablet. They then tactually explored all four letters in normal or mirror-reversed form at different rotations (0°, 90°, 180°, and 270°) and indicated letter form (i.e., normal or mirror-reversed) by pressing one of two mouse buttons. We observed the typical effect of rotation angle on object discrimination performance (i.e., greater deviation from 0° resulted in worse performance) for trained letters, consistent with mental rotation of these haptically-rendered objects. We likewise observed generally slower and less accurate performance with mirror-reversed compared to prototypically oriented stimuli. Our findings extend existing research in multisensory object recognition by indicating that a new technology simulating active haptic feedback can support the generation and spatial manipulation of mental representations of objects. Thus, such haptic tablets can offer a new avenue to mitigate visual impairments and train skills dependent on mental object-based representations and their spatial manipulation.


Does head-only exposure to GSM-900 electromagnetic fields affect the performance of rats in spatial learning tasks?
The rapid expansion of mobile communication has generated intense interest, but has also fuelled ongoing concerns. In both humans and animals, radiofrequency radiations are suspected to affect cognitive functions. More specifically, several studies performed in rodents have suggested that spatial learning can be impaired by electromagnetic field exposure. However, none of these previous studies have simulated the common conditions of GSM mobile phones use. This study is the first using a head-only exposure system emitting a 900-MHz GSM electromagnetic field (pulsed at 217 Hz). The two behavioural tasks that were evaluated here have been used previously to demonstrate performance deficits in spatial learning after electromagnetic field exposure: a classical radial maze elimination task and a spatial navigation task in an open-field arena (dry-land version of the Morris water maze). The performances of rats exposed for 45 min to a 900-MHz electromagnetic field (1 and 3.5 W/kg) were compared to those of sham-exposed and cage-control rats. There were no differences among exposed, sham, and cage-control rats in the two spatial learning tasks. The discussion focuses on the potential reasons that led previous studies to conclude that learning deficits do occur after electromagnetic field exposure.


Differential expression of proteins in Listeria monocytogenes under thermotolerance-inducing, heat shock, and prolonged heat shock conditions.
Listeria monocytogenes is a foodborne pathogen capable of employing stress adaptive responses to evade a variety of stressors including temperature stress. We employed two-dimensional gel electrophoresis coupled with matrix-assisted laser desorption/ionization-time of flight analysis to study the differential expression of L. monocytogenes (ATCC 43256) soluble proteins at heat shock (60 degrees C) conditions, prolonged heat shock (60 degrees C for 9 minutes) conditions, and thermotolerance-inducing (48 degrees C for 30 minutes followed by 60 degrees C for 9 minutes) conditions. We compared the proteome of L. monocytogenes under these conditions to the proteome at 37 degrees C. Eighteen proteins were differentially expressed at 60 degrees C (6 up-regulated and 12 down-regulated), 21 proteins were differentially expressed (12 up-regulated and 9 down-regulated) when the cells were exposed to 60 degrees C for 9 minutes, and 20 proteins were differentially expressed (10 up-regulated and 10 down-regulated) when cells were initially exposed to 48 degrees C for 30 minutes before 60 degrees C for 9 minutes. There was one unidentifiable protein with observed molecular weight of 50 kDa which was differentially expressed across the three temperature treatments. Thermotolerance-inducing conditions caused the up-regulation of a protein by as much as 12-fold. DnaN, a previously identified stress protein, was up-regulated almost threefold at 60 degrees C. TcsA, a lipoprotein (CD4 T cell-stimulating antigen), and Gap (glyceraldehyde-3-phosphate-dehydrogenase) were selectively expressed under prolonged heat shock conditions suggesting their potential as candidate marker proteins targets for identifying temperature-stressed L. monocytogenes cells.


Inversion of optical scattering and spectral extinction measurements to recover aerosol size spectra.
Aerosols interact with optical radiation in a complicated way, but in a way that can be described through the methods of electromagnetic theory. By applying constrained linear inversion methods one can recover an estimate for the size spectrum of aerosols in the micron and submicron size range from measured optical data. The amount of information that can be secured is increased by combining measurements of spectral extinction with measurements of optical scattering. From the combined measurements one can obtain about seven pieces of information about the particle size spectrum, provided optical parameters are measured to 2.5% accuracy. The application of linear constrained inversion on simulated noise-degraded data successfully recovers major features, such as the presence of modes, in the aerosol size distribution function.


Stage-specific functions of the small Rho GTPases Cdc42 and Rac1 for adult hippocampal neurogenesis.
The molecular mechanisms underlying the generation, maturation, and integration of new granule cells generated throughout life in the mammalian hippocampus remain poorly understood. Small Rho GTPases, such as Cdc42 and Rac1, have been implicated previously in neural stem/progenitor cell (NSPC) proliferation and neuronal maturation during embryonic development. Here we used conditional genetic deletion and virus-based loss-of-function approaches to identify temporally distinct functions for Cdc42 and Rac1 in adult hippocampal neurogenesis. We found that Cdc42 is involved in mouse NSPC proliferation, initial dendritic development, and dendritic spine maturation. In contrast, Rac1 is dispensable for early steps of neuronal development but is important for late steps of dendritic growth and spine maturation. These results establish cell-autonomous and stage-specific functions for the small Rho GTPases Cdc42 and Rac1 in the course of adult hippocampal neurogenesis.


Cochlear implants in the etiopathogenesis of glioblastoma--an interesting observation or independent finding?
Various risk factors have been implicated in the pathogenesis of glioblastomas including ionizing radiation. Recent evidence has suggested a possible association between exposure to nonionizing radiofrequency electromagnetic fields (RF-EMF) generated from mobile phones and wireless devices to cause malignant transformation of the neuroglial cells, albeit this is widely debated. In this report, we discuss the development of glioblastoma in two geopolitically unrelated patients, an elderly male from the United States and a middle-aged woman from Sweden, with long-standing cochlear implants (CI). We hypothesize that the low-frequency RF-EMF emanating from the transcutaneous link of the CI prosthesis over a long period has potentially triggered tumor development in these patients.


Sustaining global agriculture through rapid detection and deployment of genetic resistance to deadly crop diseases.
Contents Summary 45 I. Introduction 45 II. Targeted chromosome-based cloning via long-range assembly (TACCA) 46 III. Resistance gene cloning through mutational mapping (MutMap) 47 IV. Cloning through mutant chromosome sequencing (MutChromSeq) 47 V. Rapid cloning through resistance gene enrichment and sequencing (RenSeq) 49 VI. Cloning resistance genes through transcriptome profiling (RNAseq) 49 VII. Resistance gene deployment strategies 49 VIII. Conclusions 50 Acknowledgements 50 References 50 SUMMARY: Genetically encoded resistance is a major component of crop disease management. Historically, gene loci conferring resistance to pathogens have been identified through classical genetic methods. In recent years, accelerated gene cloning strategies have become available through advances in sequencing, gene capture and strategies for reducing genome complexity. Here, I describe these approaches with key emphasis on the isolation of resistance genes to the cereal crop diseases that are an ongoing threat to global food security. Rapid gene isolation enables their efficient deployment through marker-assisted selection and transgenic technology. Together with innovations in genome editing and progress in pathogen virulence studies, this creates further opportunities to engineer long-lasting resistance. These approaches will speed progress towards a future of farming using fewer pesticides.


The Ink4/Arf locus is a barrier for iPS cell reprogramming.
The mechanisms involved in the reprogramming of differentiated cells into induced pluripotent stem (iPS) cells by the three transcription factors Oct4 (also known as Pou5f1), Klf4 and Sox2 remain poorly understood. The Ink4/Arf locus comprises the Cdkn2a-Cdkn2b genes encoding three potent tumour suppressors, namely p16(Ink4a), p19(Arf) and p15(Ink4b), which are basally expressed in differentiated cells and upregulated by aberrant mitogenic signals. Here we show that the locus is completely silenced in iPS cells, as well as in embryonic stem (ES) cells, acquiring the epigenetic marks of a bivalent chromatin domain, and retaining the ability to be reactivated after differentiation. Cell culture conditions during reprogramming enhance the expression of the Ink4/Arf locus, further highlighting the importance of silencing the locus to allow proliferation and reprogramming. Indeed, the three factors together repress the Ink4/Arf locus soon after their expression and concomitant with the appearance of the first molecular markers of 'stemness'. This downregulation also occurs in cells carrying the oncoprotein large-T, which functionally inactivates the pathways regulated by the Ink4/Arf locus, thus indicating that the silencing of the locus is intrinsic to reprogramming and not the result of a selective process. Genetic inhibition of the Ink4/Arf locus has a profound positive effect on the efficiency of iPS cell generation, increasing both the kinetics of reprogramming and the number of emerging iPS cell colonies. In murine cells, Arf, rather than Ink4a, is the main barrier to reprogramming by activation of p53 (encoded by Trp53) and p21 (encoded by Cdkn1a); whereas, in human fibroblasts, INK4a is more important than ARF. Furthermore, organismal ageing upregulates the Ink4/Arf locus and, accordingly, reprogramming is less efficient in cells from old organisms, but this defect can be rescued by inhibiting the locus with a short hairpin RNA. All together, we conclude that the silencing of Ink4/Arf locus is rate-limiting for reprogramming, and its transient inhibition may significantly improve the generation of iPS cells.


Pre-eclampsia and risk of cardiovascular disease and cancer in later life: systematic review and meta-analysis.
To quantify the risk of future cardiovascular diseases, cancer, and mortality after pre-eclampsia. Systematic review and meta-analysis. Embase and Medline without language restrictions, including papers published between 1960 and December 2006, and hand searching of reference lists of relevant articles and reviews for additional reports. Prospective and retrospective cohort studies were included, providing a dataset of 3,488,160 women, with 198,252 affected by pre-eclampsia (exposure group) and 29,495 episodes of cardiovascular disease and cancer (study outcomes). After pre-eclampsia women have an increased risk of vascular disease. The relative risks (95% confidence intervals) for hypertension were 3.70 (2.70 to 5.05) after 14.1 years weighted mean follow-up, for ischaemic heart disease 2.16 (1.86 to 2.52) after 11.7 years, for stroke 1.81 (1.45 to 2.27) after 10.4 years, and for venous thromboembolism 1.79 (1.37 to 2.33) after 4.7 years. No increase in risk of any cancer was found (0.96, 0.73 to 1.27), including breast cancer (1.04, 0.78 to 1.39) 17 years after pre-eclampsia. Overall mortality after pre-eclampsia was increased: 1.49 (1.05 to 2.14) after 14.5 years. A history of pre-eclampsia should be considered when evaluating risk of cardiovascular disease in women. This association might reflect a common cause for pre-eclampsia and cardiovascular disease, or an effect of pre-eclampsia on disease development, or both. No association was found between pre-eclampsia and future cancer.


Fluorescence correlation spectroscopy for multiple-site equilibrium binding: a case of doxorubicin-DNA interaction.
Quantitative description of the interaction between doxorubicin (DOX), a broadly used anticancer drug, and DNA is the key to understand the action mechanism and side effects of its clinical use. However, the reported equilibrium constants of DOX-DNA interaction obtained using a range of different analytical methods vary even by several orders of magnitude. Herein, we propose a novel application of a single-molecule technique - fluorescence correlation spectroscopy (FCS) - to probe the interaction between DOX and two types of DNA (pUC19 and calf thymus DNA), taking advantage of intrinsic self-fluorescence of DOX. We provide an analytical formula for autocorrelation analysis to determine the equilibrium constant of DOX-DNA complex-formation, where binding of multiple DOX molecules to a DNA chain is included in the reaction-diffusion model. Our FCS-based method not only quantitatively revealed the values of equilibrium constant, but also implied that the stability of DOX-DNA complex is related to the types of base pair rather than the length or structure of the DNA. This work opens a promising pathway toward quantitative determination of molecular interactions in complex systems such as living cells or organisms at single-molecule level.


A new mitochondrial DNA mutation at 14577 T/C is probably a major pathogenic mutation for maternally inherited type 2 diabetes.
From a family of 16 diabetic patients with typical maternal inheritance, we investigated a 69-year-old woman with type 2 diabetes. The proband showed no major deletions in the mitochondrial DNA (mtDNA). Direct sequencing revealed 7 missense and 5 ribosomal RNA homoplasmic nucleotide substitutions when compared with the Cambridge Sequence and its recent revision. When compared with the control cybrid cells, the proband cybrid cells showed 6 nucleotide substitutions. Among these, 14577 T/C, which turned out to be 98.9% heteroplasmic, is a new missense substitution in the NADH dehydrogenase 6 gene. We also observed 2 other patients with 14577 T/C substitution from another group of 252 unrelated diabetic patients, whereas no individual from a group of 529 control subjects had 14577 T/C substitution. Furthermore, these 6 substitutions were in linkage disequilibrium. Mitochondrial respiratory chain complex I activity and O2 consumption rates of the proband cybrid cells, which were obtained by the fusion of mtDNA-deleted (rho0) HeLa cells and mtDNA from the proband, showed 64.5 and 61.5% reductions, respectively, compared with control cybrid cells. The present study strongly indicates that the new mtDNA mutation at 14577 T/C is probably a major pathogenic mutation for type 2 diabetes in this family.


A potentiometric indirect uric acid sensor based on ZnO nanoflakes and immobilized uricase.
In the present work zinc oxide nanoflakes (ZnO-NF) structures with a wall thickness around 50 to 100 nm were synthesized on a gold coated glass substrate using a low temperature hydrothermal method. The enzyme uricase was electrostatically immobilized in conjunction with Nafion membrane on the surface of well oriented ZnO-NFs, resulting in a sensitive, selective, stable and reproducible uric acid sensor. The electrochemical response of the ZnO-NF-based sensor vs. a Ag/AgCl reference electrode was found to be linear over a relatively wide logarithmic concentration range (500 nM to 1.5 mM). In addition, the ZnO-NF structures demonstrate vast surface area that allow high enzyme loading which results provided a higher sensitivity. The proposed ZnO-NF array-based sensor exhibited a high sensitivity of ~66 mV/ decade in test electrolyte solutions of uric acid, with fast response time. The sensor response was unaffected by normal concentrations of common interferents such as ascorbic acid, glucose, and urea.


The use of a chemistry-based profiler for covalent DNA binding in the development of chemical categories for read-across for genotoxicity.
An important molecular initiating event for genotoxicity is the ability of a compound to bind covalently with DNA. However, not all compounds that can undergo covalent binding mechanisms will result in genotoxicity. One approach to solving this problem, when in silico prediction techniques are being used, is to develop tools that allow chemicals to be grouped into categories based on their ability to bind covalently to DNA. For this analysis to take place, compounds need to be placed within categories where the trend in toxicity can be explained by simple descriptors, such as hydrophobicity. However, this can occur only when the compounds within a category are structurally and mechanistically similar. Chemistry-based profilers have the ability to screen compounds and highlight those with similar structures to a target compound, and are thus likely to act via a similar mechanism of action. Here, examples are reported to highlight how structure-based profilers can be used to form categories and hence fill data gaps. The importance of developing a well-defined and robust category is discussed in terms of both mechanisms of action and structural similarity.


The membrane attack mechanism of complement: the three polypeptide chain structure of the eigth component (C8).
The purification of human C8 in milligram quantities from outdated human serum was achieved by ammonium sulfate precipitation (37.5-50% saturation) and ion exchange column chromatography employing CM-32 cellulose and QAE-Sephadex. The yield of C8 activity ranged from 2-9%, and the average purification was 1,700-fold. Fully reduced C8 was shown by SDS polyacrylamide gel electrophoresis to have three polypeptide chains which were present in equimolor ratios: alpha, 77,000 daltons; beta, 63,000 daltons; and gamma, 13,700 daltons. C8 denaturation by SDS and urea in the absence of reducing agents revealed two noncovalently linked subunits: alpha-gamma, 99,000 daltons, and beta, 75,000 daltons.


Samarium-153 chelate localization in malignant melanoma.
153Sm, a radiolanthanide of half life 46.27 h, has a gamma emission of 0.103 MeV which is well suited to imaging, it is also a moderate energy beta emitter and tumour localization of various 153Sm chelates was evaluated in B16 murine melanoma to assess their endoradiotherapeutic potential. 153Sm was prepared from enriched 152Sm in the Australian Nuclear Science and Technology Organization reactor. 153Sm chelates were prepared from 153Sm-chloride and their chromatographic behaviour characterized. Tumour and organ uptake of 153Sm-chloride, 153Sm-citrate and the 153Sm chelates, DTPA, HEDTA, HIDA, BZ, PBH, PIH and NTA were measured at 1, 6, 24 and 48 h after intravenous administration to C57 black mice bearing either melanotic or amelanotic B16 melanoma of mean size 0.75 cm3. Histopathological examination of the tumours at each passaging assured comparability of the degree of melanogenesis and the absence of necrosis. 153Sm-chloride was immobile on chromatography and the rapid hepatic accumulation of both 153Sm-chloride and 153Sm-citrate was attributed to in vivo formation of a colloid. In contrast, 153Sm-DTPA, moving at the solvent front on chromatography, showed no reticuloendothelial accumulation in vivo and was rapidly excreted by the kidneys without tumour uptake. The other 153Sm chelates were of intermediate stability and all localized in both melanotic and amelanotic tumours, although to a significantly lesser degree than 67Ga-citrate. The relatively high 153Sm-HIDA activity in liver and 153Sm-NTA activity in bone impaired tumour definition, but on imaging of all the 153Sm chelates only 153Sm-DTPA failed to demonstrate the B16 melanoma and the best tumour delineation was obtained using 153Sm-HEDTA.


Expert and trainee determinations of rhetorical relevance in referral and consultation letters.
Referral and consultation letters ferry patients among providers, negotiating co-operative care. Our study examined how "relevance" is signalled and decoded in these letters, from the perspective of both experts and trainees in three clinical specialties. 104 letters were collected from 16 physicians representing family medicine, psychiatry and surgery. Interviews were conducted with 14 of these physicians and 13 residents from the three specialties. All documents and transcripts were analysed for emergent themes. Six rhetorical factors influenced expert physicians' decisions about what material is relevant: educational, professional, audience, system-institutional, medical-legal, and evaluative. Each specialty placed different emphasis on these factors. Trainees reported having no instruction regarding how to construct rhetorically relevant letters, and they demonstrated awareness of only three of the factors identified by experts--professional, audience and evaluative. Experts and trainees differed in their understanding and application of these three factors. This research demonstrates that six rhetorical factors influence relevance decisions in letter writing, and that experts address these factors in tacit, dynamic and discipline-specific ways. Trainees share with experts an appreciation of the rhetorical functions of referral and consultation letters, but lack a comprehensive understanding of the influential factors and do not receive instruction in them. These findings provide a framework for instruction in this domain to equip novices to meet the expectations of their professional audiences successfully.