TNF regulates chemokine induction essential for cell recruitment, granuloma formation, and clearance of mycobacterial infection.
Host immunity to mycobacterial infection is dependent on the activation of T lymphocytes and their recruitment with monocytes to form granulomas. These discrete foci of activated macrophages and lymphocytes provide a microenvironment for containing the infection. The cytokine, TNF, is essential for the formation and maintenance of granulomas, but the mechanisms by which TNF regulates these processes are unclear. We have compared the responses of TNF-deficient (TNF(-/-)) and wild-type C57BL/6 mice to infection with Mycobacterium smegmatis, a potent inducer of TNF, and virulent Mycobacterium tuberculosis to delineate the TNF-dependent and -independent components of the process. The initial clearance of M. smegmatis was TNF independent, but TNF was required for the early expression of mRNA encoding C-C and C-X-C chemokines and the initial recruitment of CD11b(+) macrophages and CD4(+) T cells to the liver during the second week of infection. Late chemokine expression and cell recruitment developed in TNF(-/-) mice associated with enhanced Th1-like T cell responses and mycobacterial clearance, but recruited leukocytes did not form tight granulomas. Infection of TNF(-/-) mice with M. tuberculosis also resulted in an initial delay in chemokine induction and cellular recruitment to the liver. Subsequently, increased mRNA expression was evident in TNF(-/-) mice, but the loosely associated lymphocytes and macrophages failed to form granulomas and prevent progressive infection. Therefore, TNF orchestrates early induction of chemokines and initial leukocyte recruitment, but has an additional role in the aggregation of leukocytes into functional granulomas capable of controlling virulent mycobacterial infection.


TNF-stimulated MAP kinase activation mediated by a Rho family GTPase signaling pathway.
The biological response to tumor necrosis factor (TNF) involves activation of MAP kinases. Here we report a mechanism of MAP kinase activation by TNF that is mediated by the Rho GTPase family members Rac/Cdc42. This signaling pathway requires Src-dependent activation of the guanosine nucleotide exchange factor Vav, activation of Rac/Cdc42, and the engagement of the Rac/Cdc42 interaction site (CRIB motif) on mixed-lineage protein kinases (MLKs). We show that this pathway is essential for full MAP kinase activation during the response to TNF. Moreover, this MLK pathway contributes to inflammation in vivo.


Therapeutic antitumor response after immunization with a recombinant adenovirus encoding a model tumor-associated antigen.
Recombinant adenovirus (rAd), deleted of critical genes that enable viral replication and replaced with genes encoding heterologous proteins, has been shown to be a safe and effective vector in gene therapy studies. To evaluate a potential role for rAd as an immunogen, we used two different replication-defective type 2 rAds encoding the model Ag, beta-galactosidase (beta-gal). To determine whether rAd elicited the kind of immune responses therapeutic in an anti-tumor setting, the beta-gal-expressing adenocarcinoma, CT26.CL25, was used. Splenocytes from BALB/c mice immunized with 1 x 10(7) infectious units (iu) of rAd demonstrated anti-beta-gal activity after in vitro culture with the relevant L(d) beta-gal peptide. Adoptive transfer of these same splenocytes produced dramatic regression of established pulmonary metastases. However, when tumor-bearing mice were treated with 1 x 10(7) iu of rAd, no reduction in established disease was observed even when rAd was given with exogenous IL-2. To increase the viral dose delivered to each animal, we used an E1-E4-deleted rAd that could be grown to much higher titers. Significant reduction occurred with 10-fold more rAd (1 x10(8) iu) was administered. Exogenous IL-2 administration with 1 x 10(8) iu of rAd resulted in augmentation of this anti-tumor effect. These findings demonstrate that when using a nonreplicating virus, the viral dose is directly related to the immune response generated. These data constitute the first reported use of rAd in the treatment of an established experimental cancer and may have implication for the treatment of human cancer.


Progress towards a core set of outcome measures in small-vessel vasculitis. Report from OMERACT 9.
The past decade has seen a substantial increase in the number and quality of clinical trials of new therapies for vasculitis, including randomized, controlled, multicenter trials that have successfully incorporated measures of disease activity and toxicity. However, because current treatment regimens for severe disease effectively induce initial remission and reduce mortality, future trials will focus on any of several goals including: (a) treatment of mild-moderate disease; (b) prevention of chronic damage; (c) reduction in treatment toxicity; or (d) more subtle differences in remission induction or maintenance. Thus, new trials will require outcome measure instruments that are more precise and are better able to detect effective treatments for different disease states and measure chronic manifestations of disease. The OMERACT Vasculitis Working Group comprises international clinical investigators with expertise in vasculitis who, since 2002, have worked collaboratively to advance the refinement of outcome measures in vasculitis, create new measures to address domains of illness not covered by current research approaches, and harmonize outcome assessment in vasculitis. The focus of the OMERACT group to date has been on outcome measures in small-vessel vasculitis with an overall goal of creating a core set of outcome measures for vasculitis, each of which fulfills the OMERACT filter of truth, discrimination, feasibility, and identifying additional domains requiring further research. This process has been informed by several ongoing projects providing data on outcomes of disease activity, disease-related damage, multidimensional health-related quality of life, and patient-reported ratings of the burden of vasculitis.


Effect of tralokinumab, an interleukin-13 neutralising monoclonal antibody, on eosinophilic airway inflammation in uncontrolled moderate-to-severe asthma (MESOS): a multicentre, double-blind, randomised, placebo-controlled phase 2 trial.
The role of interleukin 13 in airway inflammation and remodelling in asthma is unclear. Tralokinumab is a human monoclonal antibody that neutralises interleukin 13. We aimed to evaluate whether tralokinumab would have an effect on airway eosinophilic infiltration, blood and sputum eosinophil concentrations, eosinophil activation, and airway remodelling. We did a multicentre, double-blind, randomised, placebo-controlled phase 2 trial at 15 centres across the UK, Denmark, and Canada. We enrolled participants of either sex aged 18-75 years with inadequately controlled moderate-to-severe asthma for 12 months or more, requiring treatment with inhaled corticosteroids at a stable dose. We randomly assigned participants (1:1) to receive tralokinumab (300 mg) or placebo by an interactive web-based system or voice response system. Participants and study personnel were masked to treatment allocation. Both tralokinumab and placebo were administered subcutaneously every 2 weeks. The primary outcome measure was change from baseline to week 12 in bronchial biopsy eosinophil count. Secondary outcome measures included change in blood and sputum eosinophil counts. Exploratory outcomes included fractional exhaled nitric oxide (FENO) and blood IgE concentrations. Safety analyses were carried out in all participants who received study drug. This trial is registered with ClinicalTrials.gov, number NCT02449473, and with the European Clinical Trials Database, EudraCT 2015-000857-19. Between Sept 25, 2015, and June 21, 2017, 224 participants were enrolled and screened. Of these participants, 79 were randomly assigned to receive tralokinumab (n=39) or placebo (n=40). Tralokinumab did not significantly affect bronchial eosinophil count compared with placebo at week 12 (treatment effect ratio 1·43, 95% CI 0·63-3·27; p=0·39). Compared with placebo, tralokinumab did not significantly affect blood eosinophil count (treatment effect ratio 1·21, 95% CI 1·00-1·48; p=0·055) or sputum eosinophil count (0·57, 0·06-6·00; p=0·63), but FENO concentration (0·78, 0·63-0·96; p=0·023) and total blood IgE concentration (0·86, 0·77-0·97; p=0·014) were significantly reduced. 33 (85%) of 39 patients receiving tralokinumab and 32 (80%) of 40 receiving placebo reported at least one adverse event during the treatment period. No deaths in either treatment group were observed. Treatment-related adverse events occurred more frequently in the tralokinumab group than in the placebo group (11 [28%] of 39 vs seven [18%] of 40). Tralokinumab did not significantly affect eosinophilic inflammation in bronchial submucosa, blood, or sputum compared with placebo, but did reduce FENO and IgE concentrations. These results suggest interleukin 13 is not crucial for eosinophilic airway inflammation control in moderate-to-severe asthma. AstraZeneca.


Malaria and dengue: Knowledge, attitude, practice, and effect of sensitization workshop among school teachers as health educators.
Outbreaks of vector-borne diseases (VBDs) such as dengue and malaria can overwhelm health systems in resource-poor countries. Teachers can act as excellent educators by playing a key role of delivering important health education messages to school children and targeting an important health determinant - the health behavior. This study was done to assess the knowledge, attitude, and practice (KAP) of school teachers regarding VBDs and the effect of a sensitization workshop on the same. This is a cross-sectional study done on 212 school teachers to know their KAP regarding dengue and malaria. They were also exposed to an intervention workshop after the pretest, and the gain in knowledge scores was compared. The mean age of respondents was 38.5 years and 57.5% of them were females. Participants had fairly good knowledge about dengue and malaria. More than 90% participants knew that mosquito bite is responsible for dengue and malaria. Around 75% of respondents were also aware of the symptoms of these diseases. The number of participants having a low, medium, and high pretest knowledge score (<50%, 50%-74%, and ≥75%, respectively) was 6.6%, 42.5%, and 50.9%, respectively. A significant improvement was seen in the knowledge score after the intervention workshop (χ2 = 23.6, P < 0.001). Key success for mosquito-borne diseases control depends not only on services provided by Health Authority but also on knowledge, awareness, preventive practices, and early care-seeking behavior of the community. There is a need to know and improve existing knowledge and practice regarding mosquito-borne diseases and its control in community, especially with the collaboration of school teachers as health educators.


Effect of hypovolemia on forearm vascular resistance control during exercise in the heat.
To determine the influence of hypovolemia on the control of forearm vascular resistance (FVR) during dynamic exercise, we studied five physically active men during 60 min of supine cycle ergometer exercise bouts at 35 degrees C in control (normovolemic) and hypovolemic conditions. Hypovolemia was achieved by 3 days of diuretic administration and resulted in an average decrease in plasma volume of 15.9%. Relative to normovolemia, hypovolemia caused an attenuation of the progressive rise in forearm blood flow (P less than 0.05) and an increase in heart rate (P less than 0.05) during exercise. Because mean arterial blood pressure during hypovolemic exercise was well maintained, the attenuation of forearm blood flow was due entirely to a relative increase in FVR. At the onset of dynamic exercise, FVR was increased significantly in control and hypovolemic conditions by 13.2 and 27.1 units, respectively. The increase in FVR was significantly different between control and hypovolemic conditions as well. We attributed the increased vasoconstrictor bias during hypovolemia to cardiopulmonary baroreceptor unloading and/or an increased sensitivity to cardiopulmonary baroreceptor unloading. We concluded that reduced blood flow to the periphery during exercise in the hypovolemic condition was caused entirely by an increase in vascular resistance, thereby preserving arterial blood pressure and adequate perfusion to the organs requiring increased flow.


Dynamic localization of the human papillomavirus type 11 origin binding protein E2 through mitosis while in association with the spindle apparatus.
Papillomaviral DNA replicates as extrachromosomal plasmids in squamous epithelium. Viral DNA must segregate equitably into daughter cells to persist in dividing basal/parabasal cells. We have previously reported that the viral origin binding protein E2 of human papillomavirus types 11 (HPV-11), 16, and 18 colocalized with the mitotic spindles. In this study, we show the localization of the HPV-11 E2 protein to be dynamic. It colocalized with the mitotic spindles during prophase and metaphase. At anaphase, it began to migrate to the central spindle microtubules, where it remained through telophase and cytokinesis. It was additionally observed in the midbody at cytokinesis. A peptide spanning residues 285 to 308 in the carboxyl-terminal domain of HPV-11 E2 (E2C) is necessary and sufficient to confer localization on the mitotic spindles. This region is conserved in HPV-11, -16, and -18 and bovine papillomavirus type 4 (BPV-4) E2 and is also required for the respective E2C to colocalize with the mitotic spindles. The E2 protein of bovine papillomavirus type 1 is tethered to the mitotic chromosomes via the cellular protein Brd4. However, the HPV-11 E2 protein did not associate with Brd4 during mitosis. Lastly, a chimeric BPV-1 E2C containing the spindle localization domain from HPV-11 E2C gained the ability to localize to the mitotic spindles, whereas the reciprocal chimera lost the ability. We conclude that this region of HPV E2C is critical for localization with the mitotic apparatus, enabling the HPV DNA to sustain persistent infections.


The development of rediae of Fasciola hepatica in Radix natalensis subjected twice to bimiracidial exposures.
Experimental infections of Egyptian Radix natalensis with a French isolate of Fasciola hepatica (each snail was subjected twice to a bimiracidial exposure) were carried out to determine how many sporocysts grew in these snails and to study the developmental patterns of redial generations. Single-sporocyst infections were found in 69.3% (34/49) of infected snails, with equivalent numbers of normal and abnormal patterns. Snails with two- and three-sporocyst infections were 24.4% and 6.1%, respectively. In single- and two-sporocyst infections at days 42 and 56 post-exposure, the total redial burden was significantly higher in snails with a normal redial development. In two- and three-sporocyst infections, the overall maturity of rediae was delayed at days 42 and 56. The high frequency of abnormal patterns in R. natalensis (53.1% of all infected snails showed degeneration of a first mother redia) might be due to incomplete adaptation between the snail population and the parasite. The delayed redial maturity in two- and three-sporocyst infections can mainly be explained by the volume of the snail body, which would be insufficient to allow the simultaneous differentiation of most rediae over time.


Effect of labiolingual inclination of a maxillary central incisor and surrounding alveolar bone loss on periodontal stress: A finite element analysis.
The aim of this study was to investigate whether labial tooth inclination and alveolar bone loss affect the moment per unit of force (Mt/F) in controlled tipping and consequent stresses on the periodontal ligament (PDL). Three-dimensional models (n = 20) of maxillary central incisors were created with different labial inclinations (5°, 10°, 15°, and 20°) and different amounts of alveolar bone loss (0, 2, 4, and 6 mm). The Mt/F necessary for controlled tipping (Mt/Fcont) and the principal stresses on the PDL were calculated for each model separately in a finite element analysis. As labial inclination increased, Mt/Fcont and the length of the moment arm decreased. In contrast, increased alveolar bone loss caused increases in Mt/Fcont and the length of the moment arm. When Mt/F was near Mt/Fcont, increases in Mt/F caused compressive stresses to move from a predominantly labial apical region to a palatal apical position, and tensile stresses in the labial area moved from a cervical position to a mid-root position. Although controlled tipping was applied to the incisors, increases in alveolar bone loss and labial tooth inclination caused increases in maximum compressive and tensile stresses at the root apices. Increases in alveolar bone loss and labial tooth inclination caused increases in stresses that might cause root resorption at the root apex, despite the application of controlled tipping to the incisors.


Probing the interaction between solid benzene and water using vacuum ultraviolet and infrared spectroscopy.
We present results of a combined vacuum ultraviolet (VUV) and infrared (IR) photoabsorption study of amorphous benzene : water mixtures and layers to investigate the benzene-water interaction in the solid phase. VUV spectra of 1 : 1, 1 : 10 and 1 : 100 benzene : water mixtures at 24 K reveal a concentration dependent shift in the energies of the 1B2u, 1B1u and 1E1u electronic states of benzene. All the electronic bands blueshift from pure amorphous benzene towards gas phase energies with increasing water concentration. IR results reveal a strong dOH-π benzene-water interaction via the dangling OH stretch of water with the delocalised π system of the benzene molecule. Although this interaction influences the electronic states of benzene with the benzene-water interaction causing a redshift in the electronic states from that of the free benzene molecule, the benzene-benzene interaction has a more significant effect on the electronic states of benzene. VUV spectra of benzene and water layers show evidence of non-wetting between benzene and water, characterised by Rayleigh scattering tails at wavelengths greater than 220 nm. Our results also show evidence of benzene-water interaction at the benzene-water interface affecting both the benzene and the water electronic states. Annealing the mixtures and layers of benzene and water show that benzene remains trapped in/under water ice until water desorption near 160 K. These first systematic studies of binary amorphous mixtures in the VUV, supported with complementary IR studies, provide a deeper insight into the influence of intermolecular interactions on intramolecular electronic states with significant implications for our understanding of photochemical processes in more realistic astrochemical environments.


Nerve growth factor expression and release in allergic inflammatory disease of the upper airways.
It is well known that allergic airways disease is characterized by inflammation and hyperresponsiveness, but the link between these two conditions has not been elucidated. We have previously shown that in allergic rhinitis, hyperresponsiveness is attributable to increased neural reactivity. We thus hypothesized that nerve growth factor (NGF), which is expressed by inflammatory cells and effects changes that lead to increased neural responsiveness, could be a pivotal mediator in this disease. Using reverse transcription-polymerase chain reaction (RT-PCR), Western immunoblotting, and ELISA to evaluate NGF expression and release, we found that subjects with allergic rhinitis have significantly decreased NGF mRNA in superficial nasal scrapings and significantly higher baseline concentrations of NGF protein in nasal lavage fluids, compared with control subjects. Nasal provocation with allergen significantly increased NGF protein in nasal lavage fluids of subjects with allergic rhinitis, but not of control subjects. The concentrations of NGF protein in nasal lavage fluids were not affected by provocation with the vehicle for allergen or with histamine. These data provide the first evidence of a steady state of dysregulation in mucosal NGF expression and release in allergic rhinitis, and support a role of this neurotrophin in the pathophysiology of allergic inflammatory disease of the human airways.


Acinetobacter colistiniresistens sp. nov. (formerly genomic species 13 sensu Bouvet and Jeanjean and genomic species 14 sensu Tjernberg and Ursing), isolated from human infections and characterized by intrinsic resistance to polymyxins.
Strains of the genusAcinetobacter, classified as genomic species 13BJ/14TU have been previously associated with human infections and resistance to colistin. To clarify the taxonomy of this provisional group, we investigated 24 strains that have been isolated from humans since the 1960s in 10 countries. The genus-wide analysis of the rpoB and gyrB sequences of all strains and whole-genome sequences of strains representing different rpoB/gyrB genotypes showed that the 24 strains formed a distinct monophyletic group within the so-called haemolytic clade of the genus Acinetobacter. The distinctness of the group at the species level was supported by the results of the cluster analysis of the whole-cell protein fingerprints generated by matrix-assisted laser desorption ionization-time-of-flight MS. The 24 strains had very similar metabolic features and could be distinguished from other members of the genus by the combination of strong haemolytic and proteolytic activities and the ability to oxidize d-glucose and grow on phenylacetate and/or l-phenylalanine. The minimum inhibitory concentrations of the 24 strains to colistin and polymyxin B ranged from 16 to 64 mgl-1 and from 4 to 32 mgl-1, respectively, so uniformly reaching the current clinical resistance breakpoint (4 mg l-1) for these drugs. Genus-wide comparison revealed that such a consistently high level of resistance to polymyxins is a unique feature among species of the genus Acinetobacter,which occur in humans. We conclude that genomic species 13BJ/14TU represents a biologically meaningful and medically relevant species, for which the name Acinetobacter colistiniresistens sp. nov. is proposed. The type strain is NIPH 2036T (=CCM 8641T=CIP 110478T=CCUG 67966T=CNCTC 7573T).


Laser assisted extraction of pacemaker and implantable defibrillator leads.
To analyze the results of laser-assisted extraction of permanent pacemaker and defibrillator leads. We operated upon 36 patients, whose mean age was 54.2 years, and extracted 56 leads. The reasons for extracting the leads were as follows: infection in 19 patients, elective replacement in 13, and other causes in 4 patients. The mean time of catheter placement was 7.5+/-5.5 years. Forty-seven leads were from pacemakers, and 9 were from defibrillators. Thirty-eight leads were in use, 14 had been abandoned in the pacemaker pocket, and 4 had been abandoned inside the venous system. We successfully extracted 54 catheters, obtaining a 96.4% rate of success and an 82.1% rate for complete extraction. The 2 unsuccessful cases were due to the presence of calcium in the trajectory of the lead. The mean duration of laser light application was 123.0+/-104.5 s, using 5,215.2+/-4,924.0 pulses, in a total of 24.4+/-24.2 cycles of application. Thirty-four leads were extracted from the myocardium with countertraction after complete progression of the laser sheath, 12 leads came loose during the progression of the laser sheath, and the remaining 10 were extracted with other maneuvers. One patient experienced cardiac tamponade after extraction of the defibrillator lead, requiring open emergency surgery. The use of the excimer laser allowed extraction of the leads with a 96% rate of success; it was not effective in 2 patients who had calcification on the lead. One patient (2.8%) had a complication that required cardiac surgery on an emergency basis.


Direct observation of microtubule treadmilling by electron microscopy.
Using an immunoelectron microscopic procedure, we directly observed the concurrent addition and loss of chicken brain tubulin subunits from the opposite ends of microtubules containing erythrocyte tubulin domains. The polarity of growth of the brain tubulin on the ends of erythrocyte microtubules was determined to be similar to growth off the ends of Chlamydomonas axonemes. The flux rate for brain tubulin subunits in vitro was low, approximately 0.9 micron/h. Tubulin subunit flux did not continue through the entire microtubule as expected, but ceased when erythrocyte tubulin domains became exposed, resulting in a metastable configuration that persisted for at least several hours. We attribute this to differences in the critical concentrations of erythrocyte and brain tubulin. The exchange of tubulin subunits into the walls of preformed microtubules other than at their ends was also determined to be insignificant, the exchange rate being less than the sensitivity of the assay, or less than 0.2%/h.


Rise of the zombie institution, the failure of mental health nursing leadership, and mental health nursing as a zombie category.
In this paper, we propose that mental health nursing has become a zombie category, at least in the Australian context. Mental health nursing is a concept that has lost any real explanatory or conceptual power, yet nevertheless persists in public discourse and the collective imagination. In recent decades, powerful forces have contributed to the zombification of the mental health nursing workforce and the academy. An increase in medical hegemony, the ascendancy of allied health in mental health service provision, the need for uncritical and servile workers, protocol-driven work practices, and a failure of leadership to mobilize any substantial resistance to these trends have enabled the infection to spread. The recognition of zombification, active resistance against the forces that conspire to cause it, and the cultivation of genuine conscientious critical thought and debate offer the only hope of survival of mental health nursing as a thriving specialty.


Postoperative adjuvant chemotherapy with cisplatin, cyclophosphamide, and anthracycline (doxorubicin, epirubicin, pirarubicin) for endometrial cancer.
Doxorubicin and cisplatin are the most commonly used chemotherapeutic agents in the treatment of endometrial cancer, but their clinical efficacy is still controversial. The aim of this study was to retrospectively assess the efficacy and toxicity of combination chemotherapy using cisplatin, cyclophosphamide, and anthracy-clines in patients with stage III/IV adenocarcinoma of the endometrium. Forty patients with advanced endometrial cancer received postoperative adjuvant combination chemotherapy, using cisplatin (50 or 70 mg/m2), cyclophosphamide (500 mg/m2), and one of three anthracyclines (10 patients with doxorubicin [50 mg/m2], 18 with epirubicin [50 mg/m2], and 12 with pirarubicin [40 mg/m2]), from 1987 to 1999. All patients underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy, with pelvic lymph node dissection in 36 patients and paraaortic lymph node biopsy in 38 patients. Patients were considered eligible if they had adnexal metastasis, paraaortic lymph node metastasis, positive peritoneal cytology, or distant metastasis. The patients were divided into two groups: patients with no measurable lesion (group 1; n = 27), and those with residual measurable lesion (group 2; n = 13) after surgery. The response rate and progression-free survival rate were evaluated in group 2. In group 1, 7 patients (26%) had recurrence, and all of them died of the disease. No patients in stage IIIa (n = 10), however, had recurrence. In group 2, 6 of the 13 (46%) showed response to chemotherapy (complete response [CR], 31%; partial response [PR], 15%). Toxicity was moderate: 10 patients had grade 4 neutropenia; and dose reductions were mandated in 12 patients. In group 1, the survival of patients receiving chemotherapy was considered favorable, but patients with recurrent lesions had poor prognosis. On the other hand, in group 2, the efficacy of the chemotherapy was almost equal to that reported in the literature; however, this regimen did not contribute to an improvement in the survival rate. In conclusion, a new effective regimen of postoperative adjuvant therapy is highly desirable in patients with measurable residual lesions.


Palladium-catalyzed aryl-aryl cross-coupling reaction using ortho-substituted arylindium reagents.
A range of biaryl compounds (aryl-aryl, aryl-heteroaryl, and heteroaryl-heteroaryl) can be efficiently prepared by a palladium-catalyzed cross-coupling reaction between ortho-substituted triarylindium reagents and aryl halides. The triarylindium reagents are prepared by directed ortho-lithiation and transmetallation to indium from the corresponding benzene derivatives using various directed metallation groups (DMGs). The reaction proceeds smoothly in high yields and short reaction times with high atom economy (the three aryl groups attached to indium are efficiently transferred).


Standardization of patient outcomes reporting in percutaneous nephrolithotomy.
There exist no global standards for defining patient outcomes in renal stone surgery. The objective of this study was to evaluate the quality of reporting of outcomes in studies investigating percutaneous nephrolithotomy (PCNL) and to propose standardized consensus definitions for common outcomes. We performed a literature search in PubMed for randomized controlled studies that investigated PCNL between 2002 and 2012. All outcomes reported were analyzed for each study. Each article was examined to identify the definition of each reported patient outcome. Various aspects of patient outcomes were presented to a panel of 85 experts in a Delphi process consisting of three rounds. The experts were asked to select options that they believed would best describe each outcome. Finally, we composed recommendations for definition of the most common outcomes reported in PCNL studies. Eighty-three RCTs were included in the review of patient outcomes. Stone-free rate (55, 63.9%), length of stay (47, 56.6%), complication rate (44, 53.0%), and changes in hemoglobin (40, 48.2%) were the most frequently reported outcomes in randomized controlled trials of PCNL. Only 24/53 (45.3%) studies had a formal definition of stone-free status. Only 31/40 (77.5%) studies, which reported change in hemoglobin, had a unit of measurement; however, 22/40 (55.0%) did not report the timing of postoperative hemoglobin measurement. A set of recommendations for defining patient outcomes in PCNL is presented for the 15 most commonly reported outcomes in PCNL. Wide variations and underspecification exist in the definition and reporting of outcomes in PCNL. We propose recommendations for the definition of outcomes based on a review of the literature and expert opinion. Standardization of outcome definition and reporting will improve the quality of urologic research.


[Mating choice of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae): influence of male ageing on mating success].
The objective of this study was to evaluate the effect of male ageing on male pheromone release and mating success of Ceratitis capitata (Wiedemann). The effects of male ageing on mating were evaluated on five and 21 d-old males by assessing their mating success (males chosen by a female for copulation) and the amount of males releasing the sex pheromone. The mating success was evaluated by using several ratios of young to older males by increasing the number of older males:young males from 1:1 to 5:1. The mating success of the 1:1 ratio was also evaluated in field cages. The evaluation of the mating success (in the 1:1 ratio) showed a clear preference of the females for young males. Sex pheromone emission was much more common on young than older males. Even in cases were older males were more abundant (ratios 2:1 and 3:1), females still chose the young males. However, females could not distinguish young from older males in ratios of 4:1 or 5:1. Our data indicate that the ageing of C. capitata males has a considerable negative effect on their reproductive success, especially if they are found in a proportion any lower than 3:1.


Why are we failing to implement effective therapies in cardiovascular disease?
Worldwide, there are ~18 million deaths each year from cardiovascular disease and at least 2-3 times as many experience non-fatal cardiovascular events. Numerous evidence-based prevention and management guideline recommendations for cardiovascular disease are available. However, significant gaps between the evidence and its implementation persist ('evidence-practice gap'). There exist 'under-use' gaps with lack of implementation of proven effective strategies and 'over-use' gaps with inappropriate use of strategies with strong evidence against, or insufficient evidence for their effectiveness and safety. To better tackle the global burden of cardiovascular disease (CVD), more effective strategies are needed. We discuss three selected areas where advances in implementation research for CVD could provide improvements. First, a better assessment and understanding of the most important modifiable context-specific barriers to evidence-based care will allow optimal tailoring of interventions to overcome them. Second, novel community intervention strategies from outside current CVD research should be considered, especially for CVD areas where major barriers exist and little progress has been made. Examples of such interventions include cell phone text messaging, non-physician health workers for the delivery community CVD care in areas of need, and low-cost single-pill combination CVD therapy. Third, increasing our understanding of successful implementation and sustainability of improvements is essential for CVD as a widespread chronic disease. Learning how to better implement effective therapies for CVD will have a larger effect on patient outcomes than most single new drugs and is a priority for tackling the global burden of CVD.


Polypeptide-chain-elongation rate in Escherichia coli B/r as a function of growth rate.
By evaluating the kinetics of radioactive labelling of nascent and finished polypeptides, the peptide-chain elongation rate for Escherichia coli B/r at three different growth rates (mu) was determined to be 17 amino acids/s for the fast-growing cells (mu equals 1.3 and 2.0 doublings/h) and 12 amino acids/s for slow-growing cells (mu equals 0.67 doublings/h). The results agree with the growth-rate-dependence of the rate of peptide-chain elongation found for the translation of newly induced beta-galactosidase messenger in this strain and under these conditions of growth [Dalbow & Young (1975) Biochem. J. 150, 13-20]. Together with the previously observed ribosome efficiency at these growth rates [Dennis & Bremer (1974) J. Mol. Biol. 84, 407-422] the results indicate that the fraction of ribosomes engaged in protein synthesis is about 0.8 at all three growth rates.


Factors impacting complication rates for catheter ablation of atrial fibrillation from 2003 to 2015.
Complications from catheter ablation for atrial fibrillation (AF) are well described. Changing aspects of AF ablation including patient populations referred, institutional experience, and emerging catheter and pharmacological options may impact complication rates. We assessed procedural complication trends in AF ablation patients from 2003–2015 to identify what factors affect adverse event rates. We evaluated consecutively enrolled patients undergoing initial AF ablation from 2003 through 2015. Statistical analyses were performed to identify predictors of increased risk for major complications, which were defined as death, stroke, atrio-oesophageal fistula, phrenic nerve injury, cardiovascular events requiring blood transfusions or procedural interventions, or non-cardiovascular events requiring intervention. A total of 1475 patients (mean age 59.5 ± 10.5, 82% male) were evaluated. Major complications occurred in 3.9% (n = 58) of cases, including vascular access-site haematoma (1.3%), cardiac tamponade (1.1%), and cerebrovascular accident (CVA) (0.9%). Univariate analysis revealed increased risk of complications associated with hypertension (P = 0.048), CHA2DS2VASc score ≥1 (P = 0.015), and early institutional experience (P = 0.003). Populations with higher CHA2DS2VASc scores underwent AF ablation more frequently over time (P < 0.001). Novel catheters and anticoagulants did not appreciably affect complication rates. Multivariate analysis adjusting for hypertension, CHA2DS2VASc score, and institutional experience showed that higher CHA2DS2VASc score and early institutional experience were independent predictors of adverse events. Patient characteristics reflected in CHA2DS2VASc scoring and early institutional experience predict increased complication rates following AF ablation. Despite more patients with higher CHA2DS2VASc scores undergoing AF ablation, complication rates fell over time as institutional experience increased.


Endemic fluorosis of the skeleton: radiographic features in 127 patients.
A wide range of radiographic appearances have been reported in skeletal fluorosis, but little has been written about the spectrum of radiographic features. We evaluated the spectrum of radiographic appearances in this disorder to help with its diagnosis and differentiation from other metabolic skeletal disorders. One hundred twenty-seven patients with clinically proved endemic fluorosis had radiographs of the chest, spine, pelvis, elbow, forearm, and knee obtained. The radiographic findings were classified as osteosclerosis, osteopenia, intermittent growth lines, diaphyseal widening, or soft-tissue ossification. Two different osteopenic patterns were defined: an osteoporotic pattern with overall decreased bone density and an osteomalacic pattern that combines the features of osteoporosis with bone deformity. Soft-tissue ossification included involvement of ligaments, tendons, and interosseous membranes. Ninety-eight of the patients (89% of the adults) had some evidence of calcification and/or ossification of the attachments of ligaments, tendons, muscles, and interosseous membranes. Osteosclerosis was seen in 54 patients (43%), and osteopenia was seen in 51 patients (40%). Of the patients with osteopenia, the osteoporotic pattern was seen in 28 and the osteomalacic pattern in 23. Growth lines were found in 89 patients (70%). Metaphyseal osteomalacic zones were found in children. Diaphyseal widening was present in 35 patients (28%). Endemic skeletal fluorosis can have a wide variety of radiographic appearances, including calcification and/or ossification of the attachments of soft-tissue structures to bone, osteosclerosis, osteopenia, growth lines, and metaphyseal osteomalacic zones.


Identification of the renal erythropoietin-producing cells using transgenic mice.
Regulation of erythropoietin production by the kidneys is central to the control of erythropoiesis. Uncertainty about the identity of the renal cells involved has been a major obstacle to understanding this mechanism. We have used sequence from the mouse erythropoietin locus to direct expression of a marker gene, SV40 T antigen, to these cells in transgenic mice. The transgenic constructs contained an oligonucleotide marker (Epo-M) or SV40 sequence (Epo-TAg) in the 5' untranslated region of the mouse erythropoietin gene, flanked on each side by 9 and 7.5 kb of DNA from the mouse erythropoietin locus. Anemia-inducible expression of Epo-M and Epo-TAg was observed in the kidney. In one of thirteen lines, homologous integration of Epo-TAg into the mouse erythropoietin locus occurred. In transgenic mice bearing Epo-TAg at homologous and heterologous insertion sites, renal expression was restricted to a population of cells in the interstitium of the cortex and outer medulla. Immunohistochemical characterization by light and electron microscopy shows that these are the fibroblast-like type I interstitial cells.


Electrode-based approach for monitoring in situ microbial activity during subsurface bioremediation.
Current production by microorganisms colonizing subsurface electrodes and its relationship to substrate availability and microbial activity was evaluated in an aquifer undergoing bioremediation. Borehole graphite anodes were installed downgradient from a region of acetate injection designed to stimulate bioreduction of U(VI); cathodes consisted of graphite electrodes embedded at the ground surface. Significant increases in current density (< or =50 mA/m2) tracked delivery of acetate to the electrodes, dropping rapidly when acetate inputs were discontinued. An upgradient control electrode not exposed to acetate produced low, steady currents (< or =0.2 mA/m2). Elevated current was strongly correlated with uranium removal but minimal correlation existed with elevated Fe(II). Confocal laser scanning microscopy of electrodes revealed firmly attached biofilms, and analysis of 16S rRNA gene sequences indicated the electrode surfaces were dominated (67-80%) by Geobacter species. This is the first demonstration that electrodes can produce readily detectable currents despite long-range (6 m) separation of anode and cathode, and these results suggest that oxidation of acetate coupled to electron transfer to electrodes by Geobacter species was the primary source of current. Thus it is expected that current production may serve as an effective proxy for monitoring in situ microbial activity in a variety of subsurface anoxic environments.


Early appendectomy for perforated appendicitis in children should not be abandoned.
Appendectomy for the diagnosis of appendicitis was prospectively performed upon 133 children from 1 July 1982 to 30 June 1985 with no mortality. The pathologic diagnosis revealed 11 normal appendices (8.3 per cent), 54 instances of acute appendicitis (40.6 per cent), 16 instances of gangrenous appendicitis (12.0 per cent) and 52 instances of perforated appendicitis (39.1 per cent). There were no complications in those patients with normal appendices, acute appendicitis and gangrenous appendicitis and the mean length of hospitalization for these groups was 3.27, 3.37 and 4.75 days, respectively. The mean length of hospitalization for patients with perforated appendicitis was significantly longer, 9.45 days (p less than 0.05). Thirteen complications developed in 11 patients with perforated appendicitis. Eight patients required a reoperation (a major morbidity rate of 15.4 per cent). There was only one readmission which occurred three and one-half weeks after discharge for intestinal obstruction. Early appendectomy in children with a presumed diagnosis of either acute or perforated appendicitis resulted in low morbidity rates and is more cost effective than nonoperative management proposed by others due to shorter hospitalization and no planned readmission.


Epidemiology of invasive neonatal Cronobacter (Enterobacter sakazakii) infections.
About 120-150 neonatal Cronobacter spp. (Enterobacter sakazakii) infections have been described. An analysis of current case numbers, epidemiological measures and risk factors is warranted. Data of microbiologically confirmed cases, published between 2000 and 2008, have been analysed statistically. More than 100 neonatal Cronobacter infections have been reported in this period. The overall lethality of the 67 invasive infections was 26.9%. The lethality of Cronobacter meningitis, bacteraemia and necrotising enterocolitis (NEC) was calculated to be 41.9% (P < 0.0001), <10% and 19.0% (P < 0.05), respectively. Logistic regression models (P < 0.0001) revealed a higher gestational age at birth and parentage not from Europe as significant factors for a higher reporting probability of neonatal Cronobacter meningitis. Neonates with Cronobacter meningitis not originating from North America have a higher risk for lethal outcome than other neonatal Cronobacter infections (P < 0.0001). Continental differences of risk factors for Cronobacter meningitis and for the lethal outcome of neonatal meningitis should be elucidated. Neonatal Cronobacter infections are mainly associated with the contamination of infant formula and of the relevant cleaning and preparation equipment. Eleven neonatal Cronobacter infections, not caused by contaminated infant formula, have been retrieved. Other environmental sources of infection should be considered. Consistent and sufficiently informative data of invasive neonatal Cronobacter infections should be recorded in a centralized reporting system.


Rhythmic hippocampal slow oscillation characterizes REM sleep in humans.
Hippocampal rhythmic slow activity (RSA) is a well-known electrophysiological feature of exploratory behavior, spatial cognition, and rapid eye movement (REM) sleep in several mammalian species. Recently, RSA in humans during spatial navigation was reported, but systematic data regarding human REM sleep are lacking. Using mesio-temporal corticography with foramen ovale electrodes in epileptic patients, we report the presence of a 1.5-3-Hz synchronous rhythmic hippocampal oscillation seemingly specific to REM sleep. This oscillation is continuous during whole REM periods, is clearly observable by visual inspection, and appears in tonic and phasic REM sleep episodes equally. Quantitative analysis proved that this 1.5-3-Hz frequency band significantly differentiates REM sleep from waking and slow-wake sleep (SWS). No other frequency band proved to be significant or showed this high rhythmicity. Even in temporo-lateral surface recordings, although visually much less striking, the relative power of the 1.5-3-Hz frequency band differentiates REM sleep from other states with statistical significance. This could mean that the 1.5-3-Hz hippocampal RSA spreads over other cortical areas in humans as in other mammals. We suggest that this oscillation is the counterpart of the hippocampal theta of mammalian REM sleep, and that the 1.5-3-Hz delta EEG activity is a basic neurophysiological feature of human REM sleep.


Stearoyl-CoA desaturase 1 deficiency increases CTP:choline cytidylyltransferase translocation into the membrane and enhances phosphatidylcholine synthesis in liver.
Stearoyl-CoA desaturase (SCD) is the rate-limiting enzyme in monounsaturated fatty acid synthesis. Previously, we showed that Scd1 deficiency reduces liver triglyceride accumulation and considerably decreases synthesis of very low density lipoprotein and its secretion in both lean and obese mice. In the present study, we found that Scd1 deficiency significantly modulates hepatic glycerophospholipid profile. The content of phosphatidylcholine (PC) was increased by 40% and the activities of CTP:choline cytidylyltransferase (CCT), the rate-limiting enzyme in de novo PC synthesis, and choline phosphotransferase were increased by 64 and 53%, respectively, in liver of Scd1-/- mice. In contrast, the protein level of phosphatidylethanolamine N-methyltransferase, an enzyme involved in PC synthesis via methylation of phosphatidylethanolamine, was decreased by 80% in the liver of Scd1-/- mice. Membrane translocation of CCT is required for its activation. Immunoblot analyses demonstrated that twice as much CCTalpha was associated with plasma membrane in livers of Scd1-/- compared with wild type mice, suggesting that Scd1 mutation leads to an increase in CCT membrane affinity. The incorporation of [(3)H]glycerol into PC was increased by 2.5-fold in Scd1-/- primary hepatocytes compared with those of wild type mice. Furthermore, mitochondrial glycerol-3-phosphate acyltransferase activity was reduced by 42% in liver of Scd1-/- mice; however, the activities of microsomal glycerol-3-phosphate acyltransferase, diacylglycerol acyltransferase, and ethanolamine phosphotransferase were not affected by Scd1 mutation. Our study revealed that SCD1 deficiency specifically increases CCT activity by promoting its translocation into membrane and enhances PC biosynthesis in liver.


MRI progression of cerebral small vessel disease and cognitive decline in patients with hypertension.
Hypertension is associated with cognitive deficits, probably because it is a major risk factor for the development of white matter hyperintensities (WMH), lacunes, and cerebral microbleeds, which are MRI markers of cerebral small vessel disease. Studies into associations between presence or progression of these MRI markers and cognitive decline in hypertensive patients are rare. We investigated the association of baseline presence and progression of MRI markers of cerebral small vessel disease with cognitive decline over 4 years in patients with hypertension. In this longitudinal study, hypertensive patients underwent neuropsychological assessments and brain MRI at baseline and after 4 years. Presence and progression of periventricular and subcortical WMH, lacunes, and cerebral microbleeds were visually rated. In total, 128 hypertensive patients (90 patients with essential hypertension and 38 hypertensive lacunar stroke patients), mean age: 58.6 ± 12.2 years, were included. Progression of periventricular WMH was associated with cognitive decline in simple regression analysis (P = 0.001) and in multivariable analysis with correction for baseline WMH presence and potential confounders (P = 0.004). In this multivariable analysis, R of progression of periventricular WMH was 5.6%, whereas R of baseline presence of periventricular WMH was 0.6%. We did not find significant associations between baseline presence or progression of the other MRI markers and cognitive decline. In patients with hypertension, progression of periventricular WMH over 4 years is associated with cognitive decline, whereas we could not show an association between baseline periventricular WMH and cognitive decline. These results emphasize the importance of preventing progression of WMH in hypertensive patients.


A comparison of functional magnetic resonance imaging findings in children with and without a history of early exposure to general anesthesia.
Functional magnetic resonance imaging (fMRI) has been used to evaluate the long-term consequences of early exposure to neurotoxic agents. fMRI shows that different patterns of brain activation occur in ethanol-exposed subjects performing a go/no-go response inhibition task. Pharmacologically, ethanol and general anesthetics have similar receptor-level activity in the brain. This study utilizes fMRI to examine brain activation patterns in children exposed to general anesthesia and surgery during early brain development. After obtaining Nationwide Children's Hospital IRB approval, a surgical database was utilized to identify children aged 10-17 years with a history of at least 1 h of exposure to general anesthetics and surgery when they were between 0 and 24 months of age. Age- and gender-matched children without anesthesia exposure were recruited as a control group. All subjects were scanned while being presented with a go/no-go response inhibition task. Reaction time and accuracy data were acquired, and the blood-oxygen-level-dependent (BOLD) fMRI signal was measured as a biomarker for regional neuronal activity. There were no differences in terms of performance accuracy and response time. The analysis did not reveal any significant activation differences in the primary region of interest (prefrontal cortex and caudate nucleus); however, activation differences were seen in other structures, including the cerebellum, cingulate gyrus, and paracentral lobule. Early anesthetic exposure and surgery did not affect accuracy, response time, or activation patterns in the primary region of interest during performance of the task. Intergroup differences in activation patterns in other areas of the brain were observed, and the significance of these findings is unknown. fMRI appears to be a useful tool in evaluating the long-term effects of early exposure to general anesthesia.


Characterization of the Salmonella typhimurium pagC/pagD chromosomal region.
The PhoP/PhoQ two-component system regulates Salmonella typhimurium genes that are essential to bacterial virulence and survival within macrophages. The best characterized of these PhoP-activated genes (pag) is pagC, which encodes a 188-amino-acid envelope protein (W. S. Pulkkinen and S. I. Miller, J. Bacteriol. 173:86-93, 1991). We here report the identification of four genes (pagD, envE, msgA, and envF) located 5' to pagC. Each gene is transcribed from its own promoter, two of which (msgA and pagD) were defined by primer extension analysis. Three of these genes (pagD, envE, and envF) are predicted to encode envelope proteins. The pagD gene is transcribed in a direction opposite from that of and adjacent to pagC and is positively regulated by PhoP/PhoQ. Transposon insertions within pagD and msgA attenuate bacterial virulence and survival within macrophages; however, deletion of pagD has no effect on virulence. The product of the envF gene is predicted to be a lipoprotein on the basis of the presence of a consensus lipid attachment site. The low G + C content of these genes and the homology of msgA to Shigella plasmid DNA suggest that this region may have been acquired by horizontal transmission.


[Establishment of a rapid identification of adverse drug reaction program in R language implementation based on monitoring data].
To establish a clinically applicable model of rapid identification of adverse drug reaction program (RiADP) for risk management and decision-making of clinical drug use. Based on the theory of disproportion analysis, frequency method and Bayes method, a clinically applicable RiADP model in R language background was established, and the parameters of the model were interpreted by MedDRA coding. Based on the actual monitoring data of FDA, the model was validated by the assessing hepatotoxicity of lopinavir/ritonavir (LPV/r). The established RiADP model included four parameters: standard value of adverse drug reaction signal information, empirical Bayesian geometric mean value, ratio of reporting ratio and number of adverse drug reaction cases. Through the application of R language parameter package "phViD", the model parameters could be output quickly. After being encoded by MedDRA, it was converted into clinical terms to form a clinical interpretation report of adverse drug reactions. In addition, the evaluation results of LPV/r hepatotoxicity by the model were matched with the results reported in latest literature, which also proved the reliability of the model results. In this study, a rapid identification method of adverse reactions based on post marketing drug monitoring data was established in R language environment, which is capable of sending rapid warning of adverse reactions of target drugs in public health emergencies, and providing intuitive evidence for risk management and decision-making of clinical drugs.


The microRNA pathway regulates the temporal pattern of Notch signaling in Drosophila follicle cells.
Multicellular development requires the correct spatial and temporal regulation of cell division and differentiation. These processes are frequently coordinated by the activities of various signaling pathways such as Notch signaling. From a screen for modifiers of Notch signaling in Drosophila we have identified the RNA helicase Belle, a recently described component of the RNA interference pathway, as an important regulator of the timing of Notch activity in follicle cells. We found that loss of Belle delays activation of Notch signaling, which results in delayed follicle cell differentiation and defects in the cell cycle. Because mutations in well-characterized microRNA components phenocopied the Notch defects observed in belle mutants, Belle might be functioning in the microRNA pathway in follicle cells. The effect of loss of microRNAs on Notch signaling occurs upstream of Notch cleavage, as expression of the constitutively active intracellular domain of Notch in microRNA-defective cells restored proper activation of Notch. Furthermore, we present evidence that the Notch ligand Delta is an important target of microRNA regulation in follicle cells and regulates the timing of Notch activation through cis inhibition of Notch. Here we have uncovered a complex regulatory process in which the microRNA pathway promotes Notch activation by repressing Delta-mediated inhibition of Notch in follicle cells.


Antibacterial activity of red and white wine against oral streptococci.
Wine contains a number of biologically active compounds with beneficial effects on human health. The antibacterial action of commercial red and white wines against oral streptococci responsible for caries development and against S. pyogenes responsible for pharyngitis was studied. Its postcontact effect against S. mutans was also studied. Both wines displayed activity. The compounds responsible for such activities were succinic, malic, lactic, tartaric, citric, and acetic acid. The synthetic mixtures of the organic acids tested at the concentrations found in wine had greater antibacterial activity than the beverages, indicating that in wine they are inhibited by other components. Wine polyphenols displayed no activity against oral streptococci or S. pyogenes. Findings show that wine is active against oral streptococci and S. pyogenes and suggest that it enhances oral health.


Calcium intake in elderly patients with hip fractures.
Dietary calcium intake is assumed important in the prevention and treatment of osteoporosis. However, people in countries with a high calcium intake from commodities such as milk and milk products have a high incidence of hip fracture. The effect and influence of calcium intake in the prevention of osteoporotic fracture vary from different studies. To investigate premorbid daily calcium intake in patients with low energy hip fractures during four consecutive years. In total 120 patients (mean age 78+/-8.5 (SD) years) were included between 2002 and 2005. The patients answered a structured food frequency questionnaire (FFQ) and interviews on patients' daily calcium intake from food and supplements took place during a 6-month period before the fracture. Dual energy X-ray absorptiometry (DEXA) was performed in a subgroup of 15 patients. The mean daily calcium intake from food and supplementation was 970+/-500 mg. However, 38% of patients had an intake below the recommended 800 mg/day. There was no significant relationship between calcium intake and age, gender, bone mineral density, serum calcium or albumin, type of fracture or body mass index. The mean free plasma calcium concentration was 2.3+/-0.1, i.e. within the reference limit. In 2005, 80% of the patients who underwent DEXA had manifest osteoporosis. There was a trend towards decreased calcium intake over the observation period, with a mean calcium intake below 800 mg/day in 2005. Hip fracture patients had a mean calcium intake above the recommended daily intake, as assessed by a FFQ. However, more than one-third of patients had an intake below the recommended 800 mg/day. The intake appeared to decrease over the investigated years. The relationship between calcium intake and fracture susceptibility is complex.


Transmission of influenza virus in a mammalian host is increased by PB2 amino acids 627K or 627E/701N.
Since 2003, more than 380 cases of H5N1 influenza virus infection of humans have been reported. Although the resultant disease in these cases was often severe or fatal, transmission of avian influenza viruses between humans is rare. The precise nature of the barrier blocking human-to-human spread is unknown. It is clear, however, that efficient human-to-human transmission of an antigenically novel influenza virus would result in a pandemic. Influenza viruses with changes at amino acids 627 or 701 of the PB2 protein have been isolated from human cases of highly pathogenic H5 and H7 avian influenza. Herein, we have used the guinea pig model to test the contributions of PB2 627 and 701 to mammalian transmission. To this end, viruses carrying mutations at these positions were generated in the A/Panama/2007/99 (H3N2) and A/Viet Nam/1203/04 (H5N1) backgrounds. In the context of either rPan99 or rVN1203, mutation of lysine 627 to the avian consensus residue glutamic acid was found to decrease transmission. Introduction of an asparagine at position 701, in conjunction with the K627E mutation, resulted in a phenotype more similar to that of the parental strains, suggesting that this residue can compensate for the lack of 627K in terms of increasing transmission in mammals. Thus, our data show that PB2 amino acids 627 and 701 are determinants of mammalian inter-host transmission in diverse virus backgrounds.


Association between high homocyst(e)ine and ischemic stroke due to large- and small-artery disease but not other etiologic subtypes of ischemic stroke.
Elevated plasma homocyst(e)ine may be a causal and modifiable risk factor for ischemic stroke, but the results of previous studies have been conflicting. One possible explanation is that homocyst(e)ine may only be associated with certain pathophysiological subtypes of ischemic stroke. We conducted a case-control study of 219 hospital cases with a first-ever ischemic stroke and 205 randomly selected community control subjects stratified by age, sex, and postal code. With the use of established criteria, cases of stroke were classified by etiologic subtype in a blinded fashion. The prevalence of conventional vascular risk factors, fasting plasma homocyst(e)ine levels, vitamin levels, and nucleotide 677 methylene tetrahydrofolate reductase (MTHFR) genotypes were determined in cases and controls. Increasing homocyst(e)ine was a strong and independent risk factor for ischemic stroke (adjusted OR 2.7, 95% CI 1.4 to 5.1 for a 5-micromol/L increase in fasting plasma homocyst(e)ine from 10 to 15 micromol/L). Compared with the lowest quartile, the highest quartile of homocyst(e)ine was associated with an adjusted OR of ischemic stroke of 2.2 (95% CI 1.1 to 4.2). Mean plasma homocyst(e)ine was significantly higher in cases of ischemic stroke due to large-artery disease (14.1 micromol/L, 95% CI 12.5 to 15.9, P<0.001) and small-artery disease (12.7 micromol/L, 95% CI 11. 4 to 14.1, P=0.004) compared with control subjects (10.5 micromol/L; 95% CI 10.0 to 11.0) but not in cardioembolic or other etiologic subtypes of ischemic stroke. Compared with the lowest quartile of homocyst(e)ine, the upper 3 quartiles were associated with an adjusted OR of ischemic stroke due to large-artery disease of 3.0 (95% CI 0.8 to 10.8) for the second quartile, 5.6 (95% CI 1.6 to 20) for the third quartile, and 8.7 (95% CI 2.4 to 32) for the fourth quartile (P for trend=0.0005). However, despite a clear association between the TT MTHFR genotype and elevated fasting plasma homocyst(e)ine, there was no association between MTHFR genotype and ischemic stroke or subtype of ischemic stroke. There is a strong, graded association between increasing plasma homocyst(e)ine and ischemic stroke caused by large-artery atherosclerosis and, to a much lesser extent, small-artery disease, but not cardioembolic or other etiologic subtypes of ischemic stroke. Our results are consistent with the hypothesis that the deleterious effect of high homocyst(e)ine is mediated primarily via a proatherogenic effect.


Involvement of WNT Signaling in the Regulation of Gestational Age-Dependent Umbilical Cord-Derived Mesenchymal Stem Cell Proliferation.
Mesenchymal stem cells (MSCs) are a heterogeneous cell population that is isolated initially from the bone marrow (BM) and subsequently almost all tissues including umbilical cord (UC). UC-derived MSCs (UC-MSCs) have attracted an increasing attention as a source for cell therapy against various degenerative diseases due to their vigorous proliferation and differentiation. Although the cell proliferation and differentiation of BM-derived MSCs is known to decline with age, the functional difference between preterm and term UC-MSCs is poorly characterized. In the present study, we isolated UC-MSCs from 23 infants delivered at 22-40 weeks of gestation and analyzed their gene expression and cell proliferation. Microarray analysis revealed that global gene expression in preterm UC-MSCs was distinct from term UC-MSCs. WNT signaling impacts on a variety of tissue stem cell proliferation and differentiation, and its pathway genes were enriched in differentially expressed genes between preterm and term UC-MSCs. Cell proliferation of preterm UC-MSCs was significantly enhanced compared to term UC-MSCs and counteracted by WNT signaling inhibitor XAV939. Furthermore, WNT2B expression in UC-MSCs showed a significant negative correlation with gestational age (GA). These results suggest that WNT signaling is involved in the regulation of GA-dependent UC-MSC proliferation.


REVISITING DESTRUCTION IN "THE USE OF AN OBJECT".
"The Use of an Object" (1969a) has been widely recognized as among Winnicott's great papers and has deservedly received a good deal of attention. Much of that attention has focused on the importance that the paper gives to the role of destruction in bringing about the experience of externality. Yet the nature of that destruction has too often been assumed based on Winnicott's earlier writings. In the view that follows from that, destruction is equated with the aggression that fails to destroy the object, and the experience of externality is regarded just as the result of that failure. In offering a rereading of "The Use of an Object," the author suggests that, while this aspect of aggression/destruction indeed plays an important role in the establishment of externality, it is only part of the story, and that the central contribution of "The Use of an Object" is Winnicott's attempt to offer a new theory of primitive destruction, one that provides an impulsive basis for separation/externality itself. This theory and Winnicott's ongoing attempts to develop it after "The Use of an Object" led him to rethink the very nature of the drives.


Flavonoid Glycosides from Fenugreek Seeds Regulate Glycolipid Metabolism by Improving Mitochondrial Function in 3T3-L1 Adipocytes in Vitro.
Fenugreek is a well known annual herb widely used in both medicine and food. Four flavonoid glycosides have been separated from fenugreek seeds in our previous study. In this study, the effects of the four flavonoid glycosides on regulating glycolipid metabolism and improving mitochondrial function were investigated. Isoorientin showed a very significant activity among these flavonoid glycosides. First, isoorientin decreased the accumulation of lipid droplets in 3T3-L1 preadipocytes by reducing the expression of adipokines including PPARγ, C/EBPα, and FAS. Second, isoorientin restored insulin-stimulated glucose uptake in dexamethasone-induced insulin-resistant 3T3-L1 adipocytes by reactivating Akt and AMPK. Finally, isoorientin improved mitochondrial dysfunction induced by dexamethasone in 3T3-L1 adipocytes. Isoorientin also reversed dexamethasone-induced decrease in mitochondrial membrane potential (MMP) and intracellular ATP production, reduced accumulation of intracellular reactive oxygen species (ROS), and protected mitochondrial DNA (mtDNA) from oxidative damage. At the same time, mitochondrial biogenesis is promoted. Therefore, isoorientin may be an attractive candidate as a glucose-lowering and insulin-resistance-improving agent for the treatment of diabetes.


A Review of Pharmacovigilance.
Pharmacovigilance supports safe and appropriate use of drugs. Spontaneous reporting of adverse drug reactions (ADRs) is an essential component of pharmacovigilance. However, there is significant underreporting of ADRs. Adverse drug reactions have become a major problem in developing countries. Knowledge of pharmacovigilance could form the basis for interventions aimed at improving reporting rates and decreasing ADRs.


Psychological distress in spouses of men treated for early-stage prostate carcinoma.
The authors examined levels and predictors of psychological distress in the wives of men treated for early-stage prostate carcinoma (PCa). Patients with PCa (N = 165) and spouses were interviewed to assess general and cancer-specific distress. Social and intrapersonal factors of spouses as well as clinical characteristics and quality of life of patients were assessed as potential predictors of spouses' distress. Spouses reported more cancer-specific distress than did patients (P < 0.001), but did not differ from patients in general distress. Several spouse-reported factors predicted higher spouses' distress, including less education (P < 0.005), worse marriage quality and less social support (Ps < 0.005), more negative social interaction with the patient (Ps < 0.001), lower self-esteem (Ps < 0.001), less positive coping (Ps < 0.005), searching for meaning (P < 0.001), not finding meaning (P < 0.005), and greater illness uncertainty (Ps < 0.001). Patients' bowel function and mental health also predicted greater spouses' distress (Ps < 0.005). The findings indicated that overall distress in spouses of early-stage patients with PCa was modest, and it was more likely to be predicted by psychosocial than medical factors.


Chromatin switching and transcriptional regulation in disease.
Many human diseases are the result of inappropriate changes in gene expression resulting in deleterious phenotypes of specific cells. For example, loss of expression of tumour suppressors and/or ectopic expression of oncogenes underlie many cancers, a switch from an adult to a fetal gene-expression profile in cardiac myocytes results in cardiac hypertrophy and changes in the expression of many ion channel genes leads to a phenotypic switch from contractile to proliferative smooth muscle cells in vascular diseases such as neointimal hyperplasia and atherosclerosis. Understanding the molecular mechanisms responsible for these changes in gene expression is a major goal, in order to identify novel therapeutic targets.


[Access to a III level neonatal intensive care unit in Argentina].
In Argentina information does not exist about how many newborns (NB) who need to be hospitalized in a third level neonatal intensive care unit (NICU) actually accede, not even about the evolution of those who cannot accede. To analize the characteristics of NB that required to be hospitalized in a NICU and the evolution of those who do not accede. Longitudinal, prospective and observational study. There were included NB that required hospitalization in the NICU of the Hospital Garrahan during eleven months. Every request was registered and phone calls were made to know the evolution of rejected NB. The accessibility was analyzed by bivariated and multivariated tests. 1197 NB were included in the study; 75% with severe clinical condition, being of higher frequency the cardiac, respiratory and surgical pathologies; 637 NB (53%) were accepted. The NB from other provinces (OR 2, IC95% 1.4-2.8), retinophaty of the premature (OR 40, IC95% 14-85) and surgical disease (OR 1.99, IC95% 1.4-2.7) were independent factors that increased the possibilities to access; it decreased during the winter (OR 0.56, IC95% 0.40-0.77); 56 NB died; 47 could not have access to a third level NICU in spite of presenting pathologies sensitive of treatment. This information shows the fact that is of high importance to define regional strategies that allow the efficient administration of existing health resources and the opportune access of seriously ill NB patients to reference centers.


Vaccine-elicited CD8+ T cells protect against respiratory syncytial virus strain A2-line19F-induced pathogenesis in BALB/c mice.
CD8(+) T cells may contribute to vaccines for respiratory syncytial virus (RSV). Compared to CD8(+) T cells responding to RSV infection, vaccine-elicited anti-RSV CD8(+) T cells are less well defined. We used a peptide vaccine to test the hypothesis that vaccine-elicited RSV-specific CD8(+) T cells are protective against RSV pathogenesis. BALB/c mice were treated with a mixture (previously termed TriVax) of an M2(82-90) peptide representing an immunodominant CD8 epitope, the Toll-like receptor (TLR) agonist poly(I·C), and a costimulatory anti-CD40 antibody. TriVax vaccination induced potent effector anti-RSV CD8(+) cytotoxic T lymphocytes (CTL). Mice were challenged with RSV strain A2-line19F, a model of RSV pathogenesis leading to airway mucin expression. Mice were protected against RSV infection and against RSV-induced airway mucin expression and cellular lung inflammation when challenged 6 days after vaccination. Compared to A2-line19F infection alone, TriVax vaccination followed by challenge resulted in effector CD8(+) T cells with greater cytokine expression and the more rapid appearance of RSV-specific CD8(+) T cells in the lung. When challenged 42 days after TriVax vaccination, memory CD8(+) T cells were elicited with RSV-specific tetramer responses equivalent to TriVax-induced effector CD8(+) T cells. These memory CD8(+) T cells had lower cytokine expression than effector CD8(+) T cells, and protection against A2-line19F was partial during the memory phase. We found that vaccine-elicited effector anti-RSV CD8(+) T cells protected mice against RSV infection and pathogenesis, and waning protection correlated with reduced CD8(+) T cell cytokine expression.


Validation of a clinical risk scoring system, based solely on clinical presentation, for the management of pregnancy of unknown location.
To assess a scoring system to triage women with a pregnancy of unknown location. Validation of prediction rule. Multicenter study. Women with a pregnancy of unknown location. None. Scores assigned to factors identified at clinical presentation, total score calculated to assess risk of ectopic pregnancy (EP) in women with a pregnancy of unknown location, and a proposed three-tiered clinical action plan. The cohort of 1,400 women (284 ectopic pregnancies, 759 miscarriages, and 357 intrauterine pregnancies) was more diverse than the original cohort used to develop the decision rule. The recommendations of the action plan were low risk, intermediate risk, and high risk; the recommendation based on the model score was compared with clinical diagnosis. A total of 29.4% intrauterine pregnancies were identified for less frequent follow-up observation, and 18.4% nonviable gestations were identified for more frequent follow-up observation (to rule out an ectopic pregnancy) compared with intermediate risk (i.e., monitor in current standard fashion). For a decision of possible less frequent monitoring, the specificity was 90.8% (89.0-92.6) with negative predictive value of 79.0% (76.7-81.3). For a decision of more intense follow-up observation, the specificity was 95.0% (92.7-97.2). Test characteristics using the scoring system were replicated in the diverse validation cohort. A scoring system based on symptoms at presentation has value to stratify risk and influence the intensity of outpatient surveillance for women with pregnancy of unknown location but does not serve as a diagnostic tool.


Effects of an exercise programme with people living with HIV: research in a disadvantaged setting.
This study aimed to analyse the physical health effects of a community based 10-week physical activity programme with people living with HIV. It was developed, implemented and evaluated in a disadvantaged community in South Africa. A pre-post research design was chosen. Major recruitment and adherence challenges resulted in a small sample. Among the 23 participants who took part in both baseline and final testing, compliant participants (n = 12) were compared to non-compliant participants (n = 11). Immunological (CD4, viral load), anthropometric (height, weight, skinfolds and waist to hip ratio), muscular strength (h1RM) and cardiopulmonary fitness (time on treadmill) parameters were measured. The compliant and non-compliant groups were not different at baseline. Muscular strength was the parameter most influenced by compliance with the physical activity programme (F = 4.516, p = 0.047). Weight loss and improvement in cardiopulmonary fitness were restricted by the duration of the programme, compliance and influencing factors (e.g. nutrition, medication). The increase in strength is significant and meaningful in the context, as the participants' goals were to look healthy and strong to avoid HIV related stigma. The improvements in appearance were a motivational factor, especially since the changes were made visible in a short time. Practical implications for health promotion are described. More research contextualised in disadvantaged settings is needed.


[Physical workload and maximum acceptable work time among supermarket workers in Cali, Colombia].
In order to guarantee safety and health conditions in labor the maximum physical workload that can be supported by a worker during a labor day, without causing fatigue, must be determined. The objective of this study was to determine the relationship between the physical load, expressed as relative heart rate (RHR), and the maximum acceptable work time (MAWT) in a Colombian working population (n = 30) from a tropical environment. An observational study was carried out during a typical labor day in the warehouses of a supermarket. Physiological, demographic, health and labor conditions data were collected. The resting heart rate level and the average heart rate during work were measured, and the RHR was estimated according to with the model described by Wu & Wang. Significant correlations between MAWT and RHR and other physiological variables were observed. 43% of workers didn't fulfill the MAWT. The only single variable that was associated with fulfillment was a body mass index under 18.5 Kg/m2; the other associated variables were some specific conditions of the occupational environment. Results showed a negative correlation between physiological variables and MAWT. The RHR showed its potential usefulness in the enterprise's occupational health practice. It is important for companies to diminish extended schedules and implement physical conditioning programs.


[Patients' preferences for information in health care decision-making].
A survey was carried out among patients who concurred to cardiologic services to know how patients preferred to be informed about their health status, and the demographic characteristics associated to these preferences, considering the following items: knowledge about the disease, information about different therapeutic options and decision-making. From 770 people surveyed, 738 (95.8%) answered the form completely. A trend to trust only in the doctor's knowledge to obtain information (81.7%), in wanting to know the options of treatment and express one's point of view (85.9%), and to involve the family in the decisions (63.2%) was observed. 9.6% preferred to receive the minimum necessary information or "to know nothing" about an alleged serious disease. Males tended less to request options and give opinion on the subject (or: 0.64), giving less freedom to family involvement (or: 1.31). people with a lower social and economical level claim fewer options (or: 0.48) and gave less family participation (or = 1.79). Natives from other South American countries had a minor tendency to demand for options and express their thoughts (or: 0.60); and the ones with lower education level trusted less in the doctor's knowledge (or: 1.81), demanded fewer options (or: 0.45) and chose not to know the severity of the disease (or: 0.56). the analysis of the demographical variables allowed to define preferences associated to age, sex, origin, education, religion and health status. In conclusion, although it is imperative to promote the patient's autonomy, individual preferences must be taken into account before informing and compromising the patient in decision-making about his disease.


Chronic widespread pain: a three year followup of pain distribution and risk factors.
To describe the change of pain reports over time in 3 cohorts derived from the general population: (1) no chronic pain (NCP; n = 1156); (2) chronic regional pain (CRP; n = 502); and (3) chronic widespread pain (CWP; n = 242). To identify risk factors that predict the development or persistence of chronic widespread pain. A 3-year followup from 1995 to 1998 with postal questionnaire to 2425 subjects of both sexes aged 20-74 years on the west coast of Sweden. At followup, a larger proportion of subjects with initial CRP compared to initial NCP reported CWP (16.4 and 2.2%, respectively; p < 0.001). The majority of subjects (56.9%) who primarily reported CWP remained in that group at followup, but 26.8% had changed status to CRP and 16.3% to NCP. The number of painful regions (7-12 vs 0 regions) reported at baseline was the strongest predictor for the development of CWP with an odds ratio (OR) of 12.13 (95% CI 4.47-32.88). The development of CWP was also predicted by higher age (OR = 3.13, 95% CI 1.47-6.69, age-group 59-74 years vs age-group 20-34 years), and a family history of chronic pain (OR = 1.87, 95% CI 1.14-3.07). A habit of drinking alcohol weekly (OR = 0.42, 95% CI 0.21-0.85) compared to the habit of never or seldom drinking alcohol was protective, as well as having personal social support (OR = 0.49, 95% CI 0.28-0.85). The persistence of CWP was predicted by the number of painful regions (13-18 vs 1-6 regions) at baseline (OR = 7.56, 95% CI 2.17-26.30), and being an immigrant (OR = 3.22, 95% CI 1.33-7.77). Although the overall prevalence of CWP was stable over a 3-year period there was a considerable variation on an individual basis. This variability in expressing CWP was moderately predicted by a combination of risk factors, the most important being the number of painful regions at baseline. Future research will need to show how useful the identified factors are in clinical practice and whether intervention aimed at changing these factors will improve pain outcome.


Anaerobic bacteria from hypersaline environments.
Strictly anaerobic halophiles, namely fermentative, sulfate-reducing, homoacetogenic, phototrophic, and methanogenic bacteria are involved in the oxidation of organic carbon in hypersaline environments. To date, six anaerobic fermentative genera, containing nine species, have been described. Two of them are homoacetogens. Six species belong to the family Haloanaerobiaceae, as indicated by their unique 16S rRNA oligonucleotide sequences. Desulfohalobium retbaense and Desulfovibrio halophilus represent the only two moderately halophilic sulfate reducers so far reported. Among anoxygenic phototrophic anaerobes, a few purple bacteria with optimal growth at salinities between 6 and 11% NaCl have been isolated from hypersaline habitats. They belong to the genera Rhodospirillum, Chromatium, Thiocapsa, and Ectothiorhodospira. The commonest organisms isolated so far are Chromatium salexigens, Thiocapsa halophila, and Rhodospirillum salinarum. Extremely halophilic purple bacteria have most commonly been isolated from alkaline brines and require about 20 to 25% NaCl for optimal growth. They belong to the family Ectothiorodhospiraceae. Their osmoregulation involves synthesis or uptake of compatible solutes such as glycine-betaine that accumulate in their cytoplasm. The existence of methanogens in hypersaline environments is related to the presence of noncompetitive substrates such as methylamines, which originate mainly from the breakdown of osmoregulatory amines. Methanogenesis probably does not contribute to the mineralization of carbohydrates at NaCl concentrations higher than 15%. Above this concentration, sulfate reduction is probably the main way to oxidize H2 (although at rates too low to use up all the H2 formed) and occupies a terminal function kn the degradation of carbohydrates. Three genera and five species of halophilic methylotrophic methanogens have been reported. A bloom of phototrophic bacteria in the marine salterns of Salins-de-Giraud, located on the Mediterranean French coast in the Rhone Delta, is also described.


Sinus Microanatomy and Microbiota in a Rabbit Model of Rhinosinusitis.
Background: Rabbits are useful for preclinical studies of sinusitis because of similar physiologic features to humans. The objective of this study is to develop a rabbit model of sinusitis that permits assessment of microanatomy and sampling for evaluating shifts in the sinus microbiota during the development of sinusitis and to test how the mucociliary clearance (MCC) defect might lead to dysbiosis and chronic rhinosinusitis (CRS). Methods: Generation of CRS was accomplished with an insertion of a sterile sponge into the left middle meatus of New Zealand white rabbits (n = 9) for 2 weeks. After sponge removal, 4 rabbits were observed for another 10 weeks and evaluated for CRS using endoscopy, microCT, visualization of the functional micro-anatomy by micro-optical coherence tomography (μOCT), and histopathological analysis of the sinus mucosa. Samples were taken from the left middle meatus and submitted for microbiome analysis. Results: CT demonstrated opacification of all left sinuses at 2 weeks in all rabbits (n = 9), which persisted in animals followed for another 12 weeks (n = 4). Histology at week 2 showed mostly neutrophils. On week 14, significant infiltration of plasma cells and lymphocytes was noted with increased submucosal glands compared to controls (p = 0.02). Functional microanatomy at 2 weeks showed diminished periciliary layer (PCL) depth (p < 0.0001) and mucus transport (p = 0.0044) compared to controls despite a thick mucus layer. By 12 weeks, the thickened mucus layer was resolved but PCL depletion persisted in addition to decreased ciliary beat frequency (CBF; p < 0.0001). The mucin fermenting microbes (Lactobacillales, Bacteroidales) dominated on week 2 and there was a significant shift to potential pathogens (e.g., Pseudomonas, Burkholderia) by week 14 compared to both controls and the acute phase (p < 0.05). Conclusion: We anticipate this reproducible model will provide a means for identifying underlying mechanisms of airway-surface liquid (ASL) depletion and fundamental changes in sinus microbial communities that contribute to the development of CRS. The rabbit model of sinusitis exhibited diminished PCL depth with delayed mucus transport and significant alterations and shift in the sinus microbiome during the development of chronic inflammation.


Identification of Erwinia amylovora by Growth Morphology on Agar Containing Copper Sulfate and by Capsule Staining with Lectin.
Erwinia amylovora strains formed yellow colonies on minimal agar medium MM2 containing asparagine and copper sulfate (MM2Cu), in contrast to a white morphology on minimal agar without copper salt. Additionally, the colonies were mucoid to various extents. The yellow color was characteristic for the fire blight pathogen, including strains from raspberry and from other unusual host plants, and was used to establish a novel plating technique for identification of E. amylovora. The new identification method was especially superior to semi-selective media with sucrose when natural levan-deficient strains were assayed. No growth of E. amylovora was observed for the similar medium MM1 containing 2 mM CuSO4, due to its low content of as paragine. Identification by colony morphology on MM2 agar with copper was confirmed by staining the bacterial capsules with FITC-labeled lectin from Abrus precatorious, a compound which has a high affinity for galactose residues, the main sugar in the capsular exopolysaccharide amylovoran of E. amylovora. Other plant-associated bacteria usually did not produce the typical colony morphology of E. amylovora on MM2 agar with copper. Furthermore, those cells were not stained with the Abrus lectin. Capsule staining was also observed for weakly mucoid strains of E. amylovora, but not for strains with mutations affecting amylovoran synthesis. The secretion of fluorescent compounds by Pseudomonas syringae pathovars and even growth of any other bacterial colonies adjacent to E. amylovora could interfere with the formation of typical yellow colonies on MM2Cu, which could be white in case of dense plating. After screening for white colonies on LB agar, E. amylovora was identified in extracts from Cotoneaster leaves and in bark from apple trees with fire blight symptoms by its yellow growth pattern on MM2Cu agar and by capsule staining. The proposed selective medium gives a clear signal, is easy to prepare, does not contain dyes or any compounds toxic to humans, and can also detect E. amylovora strains deficient in levan synthesis.


Cancer incidence: life table risk versus cumulative risk.
To study differences between various methods of estimating cancer risk in individuals. Information was obtained from a cancer registry. Lifetime risk was calculated using the life table method, taking competing causes of death into account and using well recognised methods such as the cumulative rate and risk. Regional cancer registry of the Comprehensive Cancer Centre IKL, located in Maastricht, The Netherlands. Patients with a first primary malignancy in 1989-90 in the catchment area of the cancer registry. The lifetime risk of developing cancer was estimated to be 34.8% for males and 30.4% for females. In males, especially, the risk estimates were considerably higher when the cumulative risk was used. The life table method is convenient for estimating the probability that a person will develop cancer during a defined period or during his or her lifetime. The other estimates overestimate the risk of the disease under investigation, especially in the very elderly.


[Trends in diabetes mellitus in Brazil: the role of the nutritional transition].
Prevalence of type 2 diabetes mellitus has increased rapidly and is expected to increase even further. In Latin America countries there is an upward trend among the younger age groups, with a major negative impact on the quality of life and burden of the disease for the health care system. Increasing rates of overweight and obesity associated with lifestyle changes and aging of the population are the main contributing factors to the increasing prevalence of type 2 diabetes. Dietary shifts in the Brazilian population (low frequency of fiber-rich foods and heavy consumption of saturated fatty acids and sugars) and sedentary lifestyle are the main risk factors for obesity, type 2 diabetes, and other chronic diseases. Diabetes prevention programs are being promoted in several countries with results showing improvement in quality of life.


Pulsed magneto-motive optical coherence tomography for remote cellular imaging.
We developed pulsed magneto-motive optical coherence tomography (PMM-OCT) to reduce environmental temperature in the measurement volume and to expand the effective magnetic field distance from a pulse source. The proposed PMM-OCT system consisted of a spectral-domain OCT system and a customarily designed electrical pulse generator. The enhanced magnetic field allowed the proposed system to be able to image magnetically labeled cells in a distance as far as 30 mm away from the pulse generator. As an easy and sensitive approach, our PMM-OCT may be beneficially applied to a molecular-level imaging systems.


Filamin depletion blocks endoplasmic spreading and destabilizes force-bearing adhesions.
Cell motility is an essential process that depends on a coherent, cross-linked actin cytoskeleton that physically coordinates the actions of numerous structural and signaling molecules. The actin cross-linking protein, filamin (Fln), has been implicated in the support of three-dimensional cortical actin networks capable of both maintaining cellular integrity and withstanding large forces. Although numerous studies have examined cells lacking one of the multiple Fln isoforms, compensatory mechanisms can mask novel phenotypes only observable by further Fln depletion. Indeed, shRNA-mediated knockdown of FlnA in FlnB(-/-) mouse embryonic fibroblasts (MEFs) causes a novel endoplasmic spreading deficiency as detected by endoplasmic reticulum markers. Microtubule (MT) extension rates are also decreased but not by peripheral actin flow, because this is also decreased in the Fln-depleted system. Additionally, Fln-depleted MEFs exhibit decreased adhesion stability that appears in increased ruffling of the cell edge, reduced adhesion size, transient traction forces, and decreased stress fibers. FlnA(-/-) MEFs, but not FlnB(-/-) MEFs, also show a moderate defect in endoplasm spreading, characterized by initial extension followed by abrupt retractions and stress fiber fracture. FlnA localizes to actin linkages surrounding the endoplasm, adhesions, and stress fibers. Thus we suggest that Flns have a major role in the maintenance of actin-based mechanical linkages that enable endoplasmic spreading and MT extension as well as sustained traction forces and mature focal adhesions.


Therapeutic applications of macrophage colony-stimulating factor-1 (CSF-1) and antagonists of CSF-1 receptor (CSF-1R) signaling.
Macrophage-colony stimulating factor (CSF-1) signaling through its receptor (CSF-1R) promotes the differentiation of myeloid progenitors into heterogeneous populations of monocytes, macrophages, dendritic cells, and bone-resorbing osteoclasts. In the periphery, CSF-1 regulates the migration, proliferation, function, and survival of macrophages, which function at multiple levels within the innate and adaptive immune systems. Macrophage populations elicited by CSF-1 are associated with, and exacerbate, a broad spectrum of pathologies, including cancer, inflammation, and bone disease. Conversely, macrophages can also contribute to immunosuppression, disease resolution, and tissue repair. Recombinant CSF-1, antibodies against the ligand and the receptor, and specific inhibitors of CSF-1R kinase activity have been each been tested in a range of animal models and in some cases, in patients. This review examines the potential clinical uses of modulators of the CSF-1/CSF-1R system. We conclude that CSF-1 promotes a resident-type macrophage phenotype. As a treatment, CSF-1 has therapeutic potential in tissue repair. Conversely, inhibition of CSF-1R is unlikely to be effective in inflammatory disease but may have utility in cancer.


Renormalized coupled-cluster methods exploiting left eigenstates of the similarity-transformed Hamiltonian.
Completely renormalized (CR) coupled-cluster (CC) approaches, such as CR-CCSD(T), in which one corrects the standard CC singles and doubles (CCSD) energy for the effects of triply (T) and other higher-than-doubly excited clusters [K. Kowalski and P. Piecuch, J. Chem. Phys. 113, 18 (2000)], are reformulated in terms of the left eigenstates Phimid R:L of the similarity-transformed Hamiltonian of CC theory. The resulting CR-CCSD(T)(L) or CR-CC(2,3) and other CR-CC(L) methods are derived from the new biorthogonal form of the method of moments of CC equations (MMCC) in which, in analogy to the original MMCC theory, one focuses on the noniterative corrections to standard CC energies that recover the exact, full configuration-interaction energies. One of the advantages of the biorthogonal MMCC theory, which will be further analyzed and extended to excited states in a separate paper, is a rigorous size extensivity of the basic ground-state CR-CC(L) approximations that result from it, which was slightly violated by the original CR-CCSD(T) and CR-CCSD(TQ) approaches. This includes the CR-CCSD(T)(L) or CR-CC(2,3) method discussed in this paper, in which one corrects the CCSD energy by the relatively inexpensive noniterative correction due to triples. Test calculations for bond breaking in HF, F(2), and H(2)O indicate that the noniterative CR-CCSD(T)(L) or CR-CC(2,3) approximation is very competitive with the standard CCSD(T) theory for nondegenerate closed-shell states, while being practically as accurate as the full CC approach with singles, doubles, and triples in the bond-breaking region. Calculations of the activation enthalpy for the thermal isomerizations of cyclopropane involving the trimethylene biradical as a transition state show that the noniterative CR-CCSD(T)(L) approximation is capable of providing activation enthalpies which perfectly agree with experiment.


Microfluidic-based mini-metagenomics enables discovery of novel microbial lineages from complex environmental samples.
Metagenomics and single-cell genomics have enabled genome discovery from unknown branches of life. However, extracting novel genomes from complex mixtures of metagenomic data can still be challenging and represents an ill-posed problem which is generally approached with ad hoc methods. Here we present a microfluidic-based mini-metagenomic method which offers a statistically rigorous approach to extract novel microbial genomes while preserving single-cell resolution. We used this approach to analyze two hot spring samples from Yellowstone National Park and extracted 29 new genomes, including three deeply branching lineages. The single-cell resolution enabled accurate quantification of genome function and abundance, down to 1% in relative abundance. Our analyses of genome level SNP distributions also revealed low to moderate environmental selection. The scale, resolution, and statistical power of microfluidic-based mini-metagenomics make it a powerful tool to dissect the genomic structure of microbial communities while effectively preserving the fundamental unit of biology, the single cell.