Alternative medicine use in HIV-positive men and women: demographics, utilization patterns and health status.
Between 1995 and 1997, 1,675 HIV-positive men and women using complementary and alternative medicine (CAM) were enrolled into the Bastyr University AIDS Research Center's Alternative Medicine Care Outcomes in AIDS (AMCOA) study. Funded by the National Institutes of Health (NIH) Office of Alternative Medicine (OAM) and National Institute of Allergy and Infectious Diseases (NIAID), the AMCOA study collected information on participant demographics, health status and use of conventional and CAM therapies. Participants from 46 states completed a baseline questionnaire, while additional clinical information (such as CD4 count and HIV-RNA viral load) was obtained from laboratory records. AMCOA participants reported using more than 1,600 different types of CAM therapies (1,210 CAM substances, 282 CAM therapeutic activities and 119 CAM provider types) for treating HIV/AIDS. Approximately two-thirds (63% n = 1,054) of the AMCOA cohort reported using antiretroviral drug therapy (ART) during the six-months previous to completing the baseline questionnaire, while 37% (n = 621) indicated they were not using ART. Of those not using ART, 104 subjects reported never having used any conventional medications for their HIV and 12 subjects used only non-prescription diarrhoea medications. The most frequently reported CAM substances were vitamin C (63%), multiple vitamin and mineral supplements (54%), vitamin E (53%) and garlic (53%). CAM provider types most commonly consulted by the AMCOA cohort were massage therapists (49%), acupuncturists (45%), nutritionists (37%) and psychotherapists (35%). CAM activities most commonly used were aerobic exercise (63%), prayer (58%), massage (53%) and meditation (46%). The choice of CAM therapies among the AMCOA cohort does not appear to be solely based on scientific evidence of efficacy of individual therapies. The majority of AMCOA subjects could be characterized as using integrated medicine, since an overwhelming proportion of the cohort consult with both conventional and CAM providers and use both conventional and CAM medications, yet few subjects reported that their conventional and CAM providers work as a team. These data and this cohort set the stage for conducting studies of health status changes associated with specific CAM therapies.


Results of a randomized prospective trial of intraaortic balloon counterpulsation and intravenous nitroglycerin in patients with acute myocardial infarction.
A randomized prospective clinical trial compared combined treatment with intraaortic balloon pumping and intravenous nitroglycerin for 4 to 5 days with routine clinical management in 20 patients with extensive myocardium at risk for infarction as evidenced by a thallium defect score of 7.0 units or greater. No significant differences in mortality or clinical outcome were observed between the 10 patients receiving the combined treatment and the 10 receiving routine management. In 14 patients two-dimensional echocardiograms obtained 6 to 24 hours after the onset of symptoms and at follow-up 6 to 16 days later (after completion of combined intraaortic balloon pumping plus nitroglycerin therapy) were analyzed to determine whether infarct segment or noninfarct segment lengths were affected by therapy. Among these 14 patients, 5 (3 receiving the combined therapy and 2 receiving routine management) demonstrated an increase in infarct segment length of greater than 1.0 cm. Mean infarct segment length increased 0.30 +/- 0.44 cm in patients receiving the combined therapy and 0.29 +/- 0.36 cm in patients on routine management (p = NS). In contrast, noninfarct segment length increased greater than 1.0 cm (mean increase 1.20 +/- 0.39) in five of seven patients on routine management but in none of 7 patients receiving intraaortic balloon pumping plus nitroglycerin therapy (mean decrease 0.22 +/- 0.20 cm) (p less than 0.05). No significant differences were noted in left ventricular ejection fraction, as measured by gated blood pool scintigraphy, or thallium perfusion defect score in a comparison of day 1 (pretreatment) with day 4 thallium or day 7 to 14 gated blood pool scintigrams. Thus, in patients with extensive myocardium at risk, it is unlikely that a reduction in mortality or a significant improvement in myocardial perfusion or ventricular function can be obtained by early intervention with intraaortic balloon pumping in combination with nitroglycerin. Although this combined therapy failed to prevent infarct segment lengthening (infarct expansion), the combined afterload-lowering effects of intraaortic balloon pumping and nitroglycerin did appear to prevent dilation or remodeling of noninfarcted segments during the first 2 weeks after acute myocardial infarction.


The CBP bromodomain and nucleosome targeting are required for Zta-directed nucleosome acetylation and transcription activation.
The Epstein-Barr virus (EBV)-encoded lytic activator Zta is a bZIP protein that can stimulate nucleosomal histone acetyltransferase (HAT) activity of the CREB binding protein (CBP) in vitro. We now show that deletion of the CBP bromo- and C/H3 domains eliminates stimulation of nucleosomal HAT activity in vitro and transcriptional coactivation by Zta in transfected cells. In contrast, acetylation of free histones was not affected by the addition of Zta or by deletions in the bromo or C/H3 domain of CBP. Zta stimulated acetylation of oligonucleosomes assembled on supercoiled DNA and dinucleosomes assembled on linear DNA, but Zta-stimulated acetylation was significantly reduced for mononucleosomes. Western blotting and amino-terminal protein sequencing indicated that all lysine residues in the H3 and H4 amino-terminal tails were acetylated by CBP and enhanced by the addition of Zta. Histone acetylation was also dependent upon the Zta basic DNA binding domain, which could not be substituted with the homologous basic region of c-Fos, indicating specificity in the bZIP domain nucleosome binding function. Finally, we show that Zta and CBP colocalize to viral immediate-early promoters in vivo and that overexpression of Zta leads to a robust increase in H3 and H4 acetylation at various regions of the EBV genome in vivo. Furthermore, deletion of the CBP bromodomain reduced stable CBP-Zta complex formation and histone acetylation at Zta-responsive viral promoters in vivo. These results suggest that activator- and bromodomain-dependent targeting to oligonucleosomal chromatin is required for stable promoter-bound complex formation and transcription activity.


Integrated mapping and sequencing of a 115 kb DNA fragment from Bacillus subtilis: sequence analysis of a 21 kb segment containing the sigL locus.
A sequence strategy which combines a low redundancy shotgun approach and directed sequencing has been elaborated. Essentially, the sequences, as well as the size of the fragments utilized for a low coverage shotgun approach, were exploited for the construction of a physical map of the region to be sequenced. The latter considerably simplified the subsequent directed sequencing steps. We report the physical mapping of a 115 kb segment which covers nearly 100 kb of the hisA-cysB region of the Bacillus subtilis chromosome and contains previously sequenced genes sigL and sacB. Sequencing and analysis of a 21305 bp segment, which includes the sigL locus, revealed 21 ORFs, apparently belonging to at least seven transcription units. This segment has a G + C content greater than 47%, compared to 43% characteristic of the flanking regions, and mainly consists of genes whose products seem to be involved in the synthesis of an exopolysaccharide. These observations leave open the possibility that the analysed fragment has been acquired through horizontal transfer.


Steroid receptor coactivator-2 expression in brain and physical associations with steroid receptors.
Estradiol and progesterone bind to their respective receptors in the hypothalamus and hippocampus to influence a variety of behavioral and physiological functions, including reproduction and cognition. Work from our lab and others has shown that the nuclear receptor coactivators, steroid receptor coactivator-1 (SRC-1) and SRC-2, are essential for efficient estrogen receptor (ER) and progestin receptor (PR) transcriptional activity in brain and for hormone-dependent behaviors. While the expression of SRC-1 in brain has been studied extensively, little is known about the expression of SRC-2 in brain. In the present studies, we found that SRC-2 was highly expressed throughout the hippocampus, amygdala and hypothalamus, including the medial preoptic area (MPOA), ventral medial nucleus (VMN), arcuate nucleus (ARC), bed nucleus of the stria terminalis, supraoptic nucleus and suprachiasmatic nucleus. In order for coactivators to function with steroid receptors, they must be expressed in the same cells. Indeed, SRC-2 and ER(alpha) were coexpressed in many cells in the MPOA, VMN and ARC, all brain regions known to be involved in female reproductive behavior and physiology. While in vitro studies indicate that SRC-2 physically associates with ER and PR, very little is known about receptor-coactivator interactions in brain. Therefore, we used pull-down assays to test the hypotheses that SRC-2 from hypothalamic and hippocampal tissue physically associate with ER and PR subtypes in a ligand-dependent manner. SRC-2 from both brain regions interacted with ER(alpha) bound to agonist, but not in the absence of ligand or in the presence of the selective ER modulator, tamoxifen. Analysis by mass spectrometry confirmed these ligand-dependent interactions between ER(alpha) and SRC-2 from brain. In dramatic contrast, SRC-2 from brain showed little to no interaction with ERbeta. Interestingly, SRC-2 from both brain regions interacted with PR-B, but not PR-A, in a ligand-dependent manner. Taken together, these findings reveal that SRC-2 is expressed in brain regions known to mediate a variety of steroid-dependent functions. Furthermore, SRC-2 is expressed in many ER(alpha) containing cells in the hypothalamus. Finally, SRC-2 from brain interacts with ER and PR in a subtype-specific manner, which may contribute to the functional differences of these steroid receptor subtypes in brain.


Task difficulty modulates brain-behavior correlations in language production and cognitive control: Behavioral and fMRI evidence from a phonological go/no-go picture-naming paradigm.
Language production and cognitive control are complex processes that involve distinct yet interacting brain networks. However, the extent to which these processes interact and their neural bases have not been thoroughly examined. Here, we investigated the neural and behavioral bases of language production and cognitive control via a phonological go/no-go picture-naming task. Naming difficulty and cognitive control demands (i.e., conflict monitoring and response inhibition) were manipulated by varying the proportion of naming trials (go trials) and inhibition trials (no-go trials) across task runs. The results demonstrated that as task demands increased, participants' behavioral performance declined (i.e., longer reaction times on naming trials, more commission errors on inhibition trials) whereas brain activation generally increased. Increased activation was found not only within the language network but also in domain-general control regions. Additionally, right superior and inferior frontal and left supramarginal gyri were sensitive to increased task difficulty during both language production and response inhibition. We also found both positive and negative brain-behavior correlations. Most notably, increased activation in sensorimotor regions, such as precentral and postcentral gyri, was associated with better behavioral performance, in both successful picture naming and successful inhibition. Moreover, comparing the strength of correlations across conditions indicated that the brain-behavior correlations in sensorimotor regions that were associated with improved performance became stronger as task demands increased. Overall, our results suggest that cognitive control demands affect language production, and that successfully coping with increases in task difficulty relies on both language-specific and domain-general cognitive control regions.


Epidemiology of rotavirus diarrhoea in Africa: a review to assess the need for rotavirus immunization.
Rapid progress towards the development of rotavirus vaccines has prompted a reassessment of the disease burden of rotavirus diarrhoea in developing countries and the possible impact of these vaccines in reducing diarrhoeal morbidity and mortality among infants and young children. We examined the epidemiology and disease burden of rotavirus diarrhoea among hospitalized and clinic patients in African countries through a review of 43 published studies of the etiology of diarrhoea. The studies were carried out from 1975 through 1992, and only those in which a sample of more than 100 patients with diarrhoea were specifically screened for rotavirus by using an established diagnostic test were included. Rotavirus was detected in a median of 24% of children hospitalized for diarrhoea and in 23% who were treated as outpatients; 38% of the hospitalized patients with rotavirus were < 6 months and 81% were < 1 year of age. Rotavirus was detected year-round in nearly every country and generally exhibited distinct seasonal peaks during the dry months. In 5 countries where rotavirus strains had been G-typed, 74% of strains were of one of the four common serotypes (G1 to G4), G1 was the predominant serotype, and 26% were non-typeable. This cumulative experience from 15 African countries suggests that rotavirus is the most important cause of severe diarrhoea in African children and that most strains in circulation today belong to common G types that are included in reassortant vaccines. Wherever large numbers of cases of rotavirus diarrhoea occur early in infancy, immunization at birth may protect the children before their first symptomatic infection.


[Anatomic and morphological studies on the initiation of somatic embryos obtained from meristematic apexes of Musa sp.
The origin of somatic embryos obtained from meristematic apexes of the Musa (AAA) clone "Gran enano" was analyzed through histological and morphological studies during the various development phases of the process. The research point out that somatic embryos developed directly from perivascular parenchyma cells of the leaves. Histological sections of globular embryos showed a radial disposition to cell and the existence of an epidermal layer that surrounds the embryo completely. When citocinine (Z or BA) was added, some embryos remained in globular stage with mild signs of enlargement but with no later development of invagination. Other's embryos reached the invagination stage; and some reached the enlargement stage with active photosynthetic tissues. However there were no to generation of complete plant regardless of additional treatment, such as "osmotic shock" or the additions of GA3--At present do not have an explanations for this results. Therefore, additional experiment should be in early, intermediate and later stages of somatic embryogenesis, in order to understand the mechanisms underlying the lack of development of plants from somatics embryos.


Role of the kinesin neck region in processive microtubule-based motility.
Kinesin is a dimeric motor protein that can move along a microtubule for several microns without releasing (termed processive movement). The two motor domains of the dimer are thought to move in a coordinated, hand-over-hand manner. A region adjacent to kinesin's motor catalytic domain (the neck) contains a coiled coil that is sufficient for motor dimerization and has been proposed to play an essential role in processive movement. Recent models have suggested that the neck enables head-to-head communication by creating a stiff connection between the two motor domains, but also may unwind during the mechanochemical cycle to allow movement to new tubulin binding sites. To test these ideas, we mutated the neck coiled coil in a 560-amino acid (aa) dimeric kinesin construct fused to green fluorescent protein (GFP), and then assayed processivity using a fluorescence microscope that can visualize single kinesin-GFP molecules moving along a microtubule. Our results show that replacing the kinesin neck coiled coil with a 28-aa residue peptide sequence that forms a highly stable coiled coil does not greatly reduce the processivity of the motor. This result argues against models in which extensive unwinding of the coiled coil is essential for movement. Furthermore, we show that deleting the neck coiled coil decreases processivity 10-fold, but surprisingly does not abolish it. We also demonstrate that processivity is increased by threefold when the neck helix is elongated by seven residues. These results indicate that structural features of the neck coiled coil, although not essential for processivity, can tune the efficiency of single molecule motility.


Selective and reversible modification of kinase cysteines with chlorofluoroacetamides.
Irreversible inhibition of disease-associated proteins with small molecules is a powerful approach for achieving increased and sustained pharmacological potency. Here, we introduce α-chlorofluoroacetamide (CFA) as a novel warhead of targeted covalent inhibitor (TCI). Despite weak intrinsic reactivity, CFA-appended quinazoline showed high reactivity toward Cys797 of epidermal growth factor receptor (EGFR). In cells, CFA-quinazoline showed higher target specificity for EGFR than the corresponding Michael acceptors in a wide concentration range (0.1-10 μM). The cysteine adduct of the CFA derivative was susceptible to hydrolysis and reversibly yielded intact thiol but was stable in solvent-sequestered ATP-binding pocket of EGFR. This environment-dependent hydrolysis can potentially reduce off-target protein modification by CFA-based drugs. Oral administration of CFA quinazoline NS-062 significantly suppressed tumor growth in a mouse xenograft model. Further, CFA-appended pyrazolopyrimidine irreversibly inhibited Bruton's tyrosine kinase with higher target specificity. These results demonstrate the utility of CFA as a new class warheads for TCI.


Expression of the extracellular matrix protein tenascin in malignant and benign ovarian tumours.
The extracellular matrix protein tenascin (TN) is overexpressed in a number of solid tumours. Thi however, is the first study to examine TN expression in ovarian tumours. TN protein was examined in froze sections of 50 human ovarian tumours by immunohistochemistry. Malignant and borderline tumours showed significantly greater incidence and intensity of stromal staining than benign tumours (P < 0.0001 and P = 0.038 respectively). Seven omental metastases were also examined and showed a strikingly similar protein distribution to their primary tumour counterparts. The expression pattern of different RNA isoforms, created by alternative splicing of the primary transcript, was identified using reverse transcription-polymerase chain reactions (RT-PCR). The smallest TN RNA splice variant (284 bp) was found in all tumours examined, while the appearance of larger molecular weight transcripts (approximately 490 and 556 bp), as major forms, was predominantly limited to malignant tumours, with 9/12 malignant tumours showing this pattern compared with 1/6 benign tumours. These data suggest that malignant ovarian tumours have increased expression of TN compared with benign tumours and this may be associated with induction of specific isoforms.


Excess risk of death from intensive care unit-acquired nosocomial bloodstream infections: a reappraisal.
Overall rates of bloodstream infection (BSI) are often used as quality indicators in intensive care units (ICUs). We investigated whether ICU-acquired BSI increased mortality (by > or = 10%) after adjustment for severity of infection at ICU admission and during the pre-BSI stay. We conducted a matched, risk-adjusted (1:n), exposed-unexposed study of patients with stays longer than 72 h in 12 ICUs randomly selected from the Outcomerea database. Patients with BSI after the third ICU day (exposed group) were matched on the basis of risk-exposure time and mortality predicted at admission using the Three-Day Recalibrated ICU Outcome (TRIO) score to patients without BSI (unexposed group). Severity was assessed daily using the Logistic Organ Dysfunction (LOD) score. Of 3247 patients with ICU stays of >3 days, 232 experienced BSI by day 30 (incidence, 6.8 cases per 100 admissions); among them, 226 patients were matched to 1023 unexposed patients. Crude hospital mortality was 61.5% among exposed and 36.7% among unexposed patients (P<.0001). Attributable mortality was 24.8%. The only variable associated with both BSI and hospital mortality was the LOD score determined 4 days before onset of BSI (odds ratio [OR], 1.10; 95% confidence interval [CI], 1.03-1.16; P = .0025). The adjusted OR for hospital mortality among exposed patients (OR, 3.20; 95% CI, 2.30-4.43) decreased when the LOD score determined 4 days before onset of BSI was taken into account (OR, 3.02; 95% CI, 2.17-4.22; P<.0001). The estimated risk of death from BSI varied considerably according to the source and resistance of organisms, time to onset, and appropriateness of treatment. When adjusted for risk-exposure time and severity at admission and during the ICU stay, BSI was associated with a 3-fold increase in mortality, but considerable variation occurred across BSI subgroups. Focusing on BSI subgroups may be valuable for assessing quality of care in ICUs.


The baseline serum lipoprotein profile is related to plant stanol induced changes in serum lipoprotein cholesterol and triacylglycerol concentrations.
Baseline characteristics of subjects might be related to the effect of plant stanols on the serum lipoprotein profile. The aim of the study was to examine effects of subjects' baseline characteristics (baseline serum concentrations of lipids and lipoproteins at the start of the study, lathosterol, campesterol and sitosterol; gender, age, BMI, smoking, use of oral contraceptives and menopause) on the effects of plant stanol esters on the serum lipoprotein profile. We used data of five studies performed at our Department. A random intercept model was used for statistical analysis, using serum lipid and lipoprotein concentrations after plant stanol ester consumption, as dependent variables. After plant stanol ester consumption, higher baseline serum concentrations of total and LDL cholesterol resulted in larger absolute decreases in their respective serum concentrations. For the ratio of total to HDL cholesterol and triacylglycerol, higher baseline serum levels resulted in larger absolute and relative decreases in their serum levels. HDL cholesterol concentrations increased in subjects with low baseline concentrations and decreased in those with high baseline concentrations. Effects however were small. No relationships were observed with baseline serum cholesterol-standardized lathosterol and campesterol concentrations, although LDL cholesterol concentrations tended to decrease more at higher baseline sitosterol concentrations. No effects of other baseline characteristics were found. People with an unfavorable serum lipid and lipoprotein profile benefit even more of plant stanols than people with a more favorable profile.


Demography of the Intertidal Fucoid Hormosira banksii: Importance of Recruitment to Local Abundance.
Canopy-forming macroalgae form the basis of diverse coastal ecosystems globally. The fucoid Hormosira banksii is often the dominant canopy-forming macroalga in the temperate intertidal of southern Australia and New Zealand, where it is commonly associated with an understory of coralline turf. Hormosira banksii is susceptible to both natural and anthropogenic disturbance and despite its abundance, few studies have examined the demography of this important species. This study determined the demographic response of H. banksii to different gradients of disturbance to both its canopy and to the understory coralline turf. We established plots in which the density of H. banksii and/or understory coralline turf was manipulated in a pulse perturbation to simulate a disturbance event. The manipulated plots contained eight treatments ranging from 100% removal of H. banksii to 100% removal of the understory coralline turf. We then measured recruitment and followed individual recruits for up to 18 months to determine growth and survivorship. We found that H. banksii recruitment was seasonally variable throughout the experiment and highest over summer, survivorship of recruits was generally high, and the species was slow-growing and long-lived. Moreover, the level of disturbance did not seem to affect recruitment, growth, or survivorship and post-recruitment mortality was independent of H. banksii density. In this system, it appears that H. banksii is a relatively long-lived perennial species whose demography is density-independent which appears to allow recovery from disturbance.


Determination of vapor-liquid equilibrium data in microfluidic segmented flows at elevated pressures using Raman spectroscopy.
A fast, noninvasive, and efficient analytical measurement strategy for the characterization of vapor-liquid equilibria (VLE) is presented, which is based on phase (state of matter) selective Raman spectroscopy in multiphase flows inside microcapillay systems (MCS). Isothermal VLE data were measured in binary and ternary mixtures composed of acetone, water, carbon dioxide or nitrogen at elevated pressures up to 10 MPa and temperatures up to 333 K. For validation, the obtained data were compared with literature data and reference measurements in a high-pressure variable volume cell. Additionally, the mixtures were investigated at temperatures and pressures where no data is available in literature to extend the high-pressure VLE database.


Enhancing the activity of protein C by mutagenesis to improve the membrane-binding site: studies related to proline-10.
Bovine and human protein C show high homology in the amino acids of their GLA domains (amino-terminal 44 residues), despite the about 10-fold higher membrane affinity of the human protein. A proposed membrane contact site and mechanism suggested that this difference was largely due to the presence of proline at position 10 of bovine protein C versus histidine at position 10 of human protein C [McDonald, J.F., Shah, A.M., Schwalbe, R.A., Kisiel, W., Dahlback, B., and Nelsestuen, G.L. (1997) Biochemistry, 36, 5120-5127]. This study examined the impact of replacing proline-10 in bovine protein C with histidine, and the reverse change in human protein C. In both cases, the protein containing proline-10 showed lower membrane affinity, about 10-fold lower for bovine protein C and 5-fold lower for human protein C. As expected, activated human protein C (hAPC) containing proline at position 10 showed 2.4-3.5-fold lower activity than wild type hAPC, depending on the assay used. Most interesting was that bovine APC containing histidine-10 displayed up to 15-fold higher activity than wild type bAPC. This demonstrated the ability to improve both membrane contact and activity by mutation. This general strategy should be applicable to other vitamin K-dependent proteins, providing opportunities to study function as well as to produce proteins that may find use as promoters and inhibitors of blood coagulation in pathological states.


Energy transfer dyads based on Nile Red.
This project was initiated to develop energy transfer dyads emitting in the 600 - 700 nm region by using Nile Red acceptors. Thus fluorescein- and BODIPY-based donors were linked to these acceptors via alkynes or triazoles. The product dyads (1 - 5) have energy transfer efficiencies of 77 - 97% in organic media.


RyR1/RyR3 chimeras reveal that multiple domains of RyR1 are involved in skeletal-type E-C coupling.
Skeletal-type E-C coupling is thought to require a direct interaction between RyR1 and the alpha(1S)-DHPR. Most available evidence suggests that the cytoplasmic II-III loop of the dihydropyridine receptor (DHPR) is the primary source of the orthograde signal. However, identification of the region(s) of RyR1 involved in bidirectional signaling with the alpha(1S)-DHPR remains elusive. To identify these regions we have designed a series of chimeric RyR cDNAs in which different segments of RyR1 were inserted into the corresponding region of RyR3 and expressed in dyspedic 1B5 myotubes. RyR3 provides a preferable background than RyR2 for defining domains essential for E-C coupling because it possesses less sequence homology to RyR1 than the RyR2 backbone used in previous studies. Our data show that two regions of RyR1 (chimera Ch-10 aa 1681-2641 and Ch-9 aa 2642-3770), were independently able to restore skeletal-type E-C coupling to RyR3. These two regions were further mapped and the critical RyR1 residues were 1924-2446 (Ch-21) and 2644-3223 (Ch-19). These results both support and refine the previous hypothesis that multiple domains of RyR1 combine to functionally interact with the DHPR during E-C coupling.


Planning, delivery, and quality assurance of intensity-modulated radiotherapy using dynamic multileaf collimator: a strategy for large-scale implementation for the treatment of carcinoma of the prostate.
To improve the local control of patients with adenocarcinoma of the prostate we have implemented intensity modulated radiation therapy (IMRT) to deliver a prescribed dose of 81 Gy. This method is based on inverse planning and the use of dynamic multileaf collimators (DMLC). Because IMRT is a new modality, a major emphasis was on the quality assurance of each component of the process and on patient safety. In this article we describe in detail our procedures and quality assurance program. Using an inverse algorithm, we have developed a treatment plan consisting five intensity-modulated (IM) photon fields that are delivered with DMLC. In the planning stage, the planner specifies the number of beams and their directions, and the desired doses for the target, the normal organs and the "overlap" regions. Then, the inverse algorithm designs intensity profiles that best meet the specified criteria. A second algorithm determines the leaf motion that would produce the designed intensity pattern and produces a DMLC file as input to the MLC control computer. Our quality assurance program for the planning and treatment delivery process includes the following components: 1) verification of the DMLC field boundary on localization port film, 2) verification that the leaf motion of the DMLC file produces the planned dose distribution (with an independent calculation), 3) comparison of dose distribution produced by DMLC in a flat phantom with that calculated by the treatment planning computer for the same experimental condition, 4) comparison of the planned leaf motions with that implemented for the treatment (as recorded on the MLC log files), 5) confirmation of the initial and final positions of the MLC for each field by a record-and-verify system, and 6) in vivo dose measurements. Using a five-field IMRT plan we have customized dose distribution to conform to and deliver 81 Gy to the PTV. In addition, in the overlap regions between the PTV and the rectum, and between the PTV and the bladder, the dose is kept within the tolerance of the respective organs. Our QA checks show acceptable agreement between the planned and the implemented leaf motions. Correspondingly, film and TLD dosimetry indicates that doses delivered agrees with the planned dose to within 2%. As of September 15, 1996, we have treated eight patients to 81 Gy with IMRT. For complex planning problems where the surrounding normal tissues place severe constraints on the prescription dose, IMRT provides a powerful and efficient solution. Given a comprehensive and rigorous quality-assurance program, the intensity-modulated fields can be efficaciously and accurately delivered using DMLC. IMRT treatment is now ready for routine implementation on a large scale in our clinic.


Surface passivity largely governs the bioaccessibility of nickel-based powder particles at human exposure conditions.
The European chemical framework REACH requires that hazards and risks posed by chemicals, including alloys and metals, are identified and proven safe for humans and the environment. Therefore, differences in bioaccessibility in terms of released metals in synthetic biological fluids (different pH (1.5-7.4) and composition) that are relevant for different human exposure routes (inhalation, ingestion, and dermal contact) have been assessed for powder particles of an alloy containing high levels of nickel (Inconel 718, 57 wt% nickel). This powder is compared with the bioaccessibility of two nickel-containing stainless steel powders (AISI 316L, 10-12% nickel) and with powders representing their main pure alloy constituents: two nickel metal powders (100% nickel), two iron metal powders and two chromium metal powders. X-ray photoelectron spectroscopy, microscopy, light scattering, and nitrogen absorption were employed for the particle and surface oxide characterization. Atomic absorption spectroscopy was used to quantify released amounts of metals in solution. Cytotoxicity (Alamar blue assay) and DNA damage (comet assay) of the Inconel powder were assessed following exposure of the human lung cell line A549, as well as its ability to generate reactive oxygen species (DCFH-DA assay). Despite its high nickel content, the Inconel alloy powder did not release any significant amounts of metals and did not induce any toxic response. It is concluded, that this is related to the high surface passivity of the Inconel powder governed by its chromium-rich surface oxide. Read-across from the pure metal constituents is hence not recommended either for this or any other passive alloy.


[The current situation with regard to nicotine addiction in Burkina Faso: tobacco supply and data from the KAB-P survey of young people in Ouagadougou].
Smoking is a worldwide phenomenon and many studies have demonstrated that tobacco use is a risk factor for morbidity and mortality. We investigated aspects of the supply of, and demand for, tobacco in Burkina Faso, with a view to helping the public authorities design strategies to combat this phenomenon. We used data on the importation of tobacco and the making of cigarettes in Burkina Faso and carried out a survey of adolescents and young adults (n = 289) with a mean age of 20.9 years. In 1997, Burkina Faso imported 1,905, 214 metric tons of tobacco of all kinds, with a CAF value of 5,808, 905,269 CFA francs. In the survey, 30.7% of those interviewed said that they smoked. These individuals gave various reasons for their use of tobacco including imitating friends (38.6%) and personal liberty (14.1%). These young people were generally aware of the risks of tobacco use, but this did not deter them from smoking. In countries like Burkina Faso, which is faced with many challenges, controlling tobacco advertising and providing ways and means to help young people to stop smoking or to deter them from starting to smoke are of major importance for future public health.


Sociometric Risk Network Structure, HIV Prevalence, and Drug Injection-Related Norms among People Who Inject Drugs (PWID) in Athens, Greece.
Background: Structural properties of sociometric networks have been associated with behaviors related to HIV transmission. Very few studies, however, have explored the correlation between sociometric network factors and drug injection-related norms. Methods: This exploratory work: (i) describes basic structural qualities of a sociometric risk network of participants in the Transmission Reduction Intervention Project (TRIP) in Athens, Greece, in the context of a large HIV outbreak among people who inject drugs (PWID); (ii) measures HIV prevalence within specific structures within the sociometric risk network of PWID in TRIP; and (iii) explores the association of structural properties of the sociometric risk network in TRIP with drug injection-related norms. Results: The sociometric risk network in TRIP consisted of a large component (n = 241, 67.8%), a few small components (n = 36, 10.1%) with 2-10 individuals each, and some isolates (n = 79, 22.2%). HIV prevalence was significantly higher in the large component (55.6%), the 2-core (59.1%) and 3-core (66.3%) of the large component, and the 3-cliques of the cores. Drug injection-related norms were significantly associated with structural characteristics of the sociometric risk network. A safe behavioral pattern (use of unclean cooker/filter/rinse water was never encouraged) was significantly (p = 0.03) less normative among people who TRIP participants of the 2-core injected with (40.5%) than among network contacts of TRIP participants outside the 2-core (55.6%). On the contrary, at drug-using venues, 2-core members reported that safer behaviors were normative compared to what was reported by those without 2-core membership. Conclusions: Sociometric network data can give useful insights into HIV transmission dynamics and inform prevention strategies.Supplemental data for this article is available online at https://doi.org/10.1080/10826084.2021.1914103 .


Trends in spending on eating away from home in Brazil, 2002-2003 to 2008-2009.
The study aims to describe trends in food consumption away from home in Brazil from 2002-2003 to 2008-2009 and to analyze the influence of income on this behavior. The authors used data collected by the Household Budget Surveys conducted by the Brazilian Institute of Geography and Statistics (IBGE) in 2002-2003 and 2008-2009. The information analyzed in this study involves records of food and beverage purchases for consumption away from home. Trends in eating away from home were estimated for the total population and according to demographic and economic strata. The association between the share of food consumed away from home and income was studied using regression models to estimate income elasticity coefficients. The share of eating away from home increased 25% during the period, reaching 28% of total spending on food. Each 10% increase in mean per capita income leads to a 3.5% increase in the share of food consumed away from home. This suggests that income growth will result in future increases in the share of eating away from home.


Modulation of the bovine delayed-type hypersensitivity responses to defined mycobacterial antigens by a synthetic bacterial lipopeptide.
The use of defined protein and peptide antigens can overcome specificity limitations of purified protein derivatives in the detection of bovine tuberculosis when the antigens are used in blood-based tests. Since the use of these specific antigens as skin test reagents could have practical advantages, we investigated the potential of Mycobacterium bovis-specific antigens to stimulate delayed-type hypersensitivity (DTH) responses in cattle experimentally infected with M. bovis. A cocktail of the recombinant antigens ESAT-6, MPB83, and MPB64 failed to stimulate in vivo DTH in cattle that had been experimentally infected with M. bovis despite the fact that the antigens were recognized in vitro by the same animals. However, it was possible to stimulate antigen-specific bovine DTH responses by using ESAT-6 in combination with a synthetic bacterial lipopeptide. This lipopeptide stimulated the release of the proinflammatory cytokine tumor necrosis factor alpha from monocyte-derived bovine dendritic cells in vitro, thereby providing a possible mechanism for its DTH-enhancing properties.


Lunar Gravity over Large Craters from Apollo 12 Tracking Data.
The Doppler residuals from the Apollo 12 lunar module radio tracking data indicate large negative accelerations over the craters Ptolemaeus and Albategnius. The mass deficienicies required to produce these accelerations are approximately equivalent to the removal of the surface material to a depth of 1 kilometer over the entire area of these craters. Several other features of the gravity fine structure can also be correlated with topography.


Efficacy of the Antibody-Drug Conjugate W0101 in Preclinical Models of IGF-1 Receptor Overexpressing Solid Tumors.
The insulin-like growth factor type 1 receptor (IGF-1R) is important in tumorigenesis, and its overexpression occurs in numerous tumor tissues. To date, therapeutic approaches based on mAbs and tyrosine kinase inhibitors targeting IGF-1R have only shown clinical benefit in specific patient populations. We report a unique IGF-1R-targeted antibody-drug conjugate (ADC), W0101, designed to deliver a highly potent cytotoxic auristatin derivative selectively to IGF-1R overexpressing tumor cells. The mAb (hz208F2-4) used to prepare the ADC was selected for its specific binding properties to IGF-1R compared with the insulin receptor, and for its internalization properties. Conjugation of a novel auristatin derivative drug linker to hz208F2-4 did not alter its binding and internalization properties. W0101 induced receptor-dependent cell cytotoxicity in vitro when applied to various cell lines overexpressing IGF-1R, but it did not affect normal cells. Efficacy studies were conducted in several mouse models expressing different levels of IGF-1R to determine the sensitivity of the tumors to W0101. W0101 induced potent tumor regression in certain mouse models. Interestingly, the potency of W0101 correlated with the expression level of IGF-1R evaluated by IHC. In an MCF-7 breast cancer model with high-level IGF-1R expression, a single injection of W0101 3 mg/kg led to strong inhibition of tumor growth. W0101 provides a potential new therapeutic option for patients overexpressing IGF-1R. A first-in-human trial of W0101 is currently ongoing to address clinical safety.


The minimally effective concentration of adrenaline in a low-concentration thoracic epidural analgesic infusion of bupivacaine, fentanyl and adrenaline after major surgery. A randomized, double-blind, dose-finding study.
We have documented that adrenaline 2.0 micro g.ml- 1 markedly improves relief of dynamic pain when added to a thoracic epidural analgesic infusion of bupivacaine 1 mg.ml- 1 and fentanyl 2 micro g.ml- 1. Concern about possible adverse effects on spinal cord blood flow, expressed by others, prompted us to find the lowest concentration of adrenaline needed to produce effective and reliable pain relief after major surgery. A prospective, randomized, double-blind, parallel group study was carried out in 36 patients after major thoracic or upper abdominal surgery. Patients with only mild pain when coughing during titrated thoracic epidural infusion of approximately 9 ml per hour of bupivacaine 1 mg.ml- 1, fentanyl 2 micro g.ml- 1, and adrenaline 2.0 micro g.ml- 1 were included. The study was conducted as a dose-finding study comparing three different adrenaline concentrations in the epidural mixture (0.5, 1.0, and 1.5 micro g.ml- 1) with each other and with adrenaline 2.0 micro g.ml- 1 in our standard epidural mixture. On the 1st postoperative day, the patients were randomly allocated into three equal groups of 12 patients each, and given a double-blind epidural infusion at the same rate, but with different adrenaline concentrations (0.5, 1.0, or 1.5 micro g.ml- 1). The effects were observed for 4 h or until pain when coughing became unacceptable in spite of rescue analgesia. Rescue analgesia consisted of up to two patient-controlled epidural bolus injections per hour (4 ml) and subsequent i.v. morphine, if necessary. All patients received rectal paracetamol 1 g, every 6th hour. Main outcome measures were pain intensity at rest and when coughing, evaluated by a visual analogue scale and an overall quality of pain relief score. The extent of sensory blockade was evaluated by determining dermatomal hypaesthesia to cold. Pain intensity when coughing increased (P < 0.001) and the number of hypaesthetic dermatomal segments decreased (P < 0.002) when the concentration of adrenaline was reduced below 1.5 micro g.ml- 1 in the triple epidural mixture. This change started within two hours after reducing the concentration of adrenaline below 1.5 micro g.ml- 1. The differences in pain intensities at rest were less pronounced. After 4 h with adrenaline 0.5 or 1.0 micro g.ml- 1 pain intensity when coughing was unacceptable in spite of rescue analgesia. After restarting the standard epidural mixture with adrenaline 2.0 micro g.ml- 1, pain intensity was again reduced to mild pain when coughing and the sensory blockade was restored. Occurrence of pruritus increased with a decreasing adrenaline concentration. Adrenaline in a dose-related manner improves the pain-relieving effect and sensory blockade and decreases the occurrence of pruritus of a low-concentration thoracic epidural analgesic infusion of bupivacaine 1 mg. ml- 1 and fentanyl 2 micro g.ml- 1 after major thoracic or upper abdominal surgery. The minimally effective concentration of adrenaline, when added to bupivacaine 1 mg.ml- 1 and fentanyl 2 micro g.ml- 1, to maintain relief of dynamic pain is approximately 1.5 micro g.ml- 1. The data clearly document that dynamic, cough-provoked pain is a more sensitive outcome measure for postoperative pain relief than pain at rest.


[Use of therapeutic plasmapheresis in heart surgery patients with hepatic insufficiency].
Therapeutic plasmapheresis is a highly effective method in the preparation of patients with hepatic insufficiency for operation on the heart under conditions of extracorporeal circulation. It leads to reduction of the bilirubin level and normalization of the main values of hepatic function. The positive effect of therapeutic plasmapheresis is less manifest in acute hepatorenal insufficiency developing after operation with extracorporeal circulation. The best effect of therapeutic plasmapheresis in the postoperative period was produced in patients in whom hemorrhages and the low cardiac output syndrome were removed.


Social support and mastery influence the association between stress and poor physical health in parents caring for children with developmental disabilities.
To date, much of the research linking the stress of caring for children with developmental disabilities (e.g. Autism & Down syndrome) with parental health outcomes have tended to concentrate on mental health with less attention paid to the physical health consequences. Thus, this study sought to explore the psychosocial predictors of poor physical health in these caring parents. One hundred and sixty-seven parents (109 caregivers and 58 control parents) completed measures of stress, child problem behaviours, social support, mastery and physical health. Parents of children with developmental disabilities had poorer physical health compared to control parents. Stress and mastery, but not social support and problem behaviours, were significant predictors of poor physical health within caring parents for children with developmental disabilities. However, the association between mastery and physical health was mediated by perceived stress such that those parents who were higher on mastery reported less stress and better physical health; furthermore, the association between stress and physical health was moderated by social support; those parents high on social support and low in stress had better physical health. These results indicate that the paths between psychosocial factors and poor physical health in the caring parents are working synergistically rather than in isolation. They also underscore the importance of providing multi-component interventions that offer a variety of psychosocial resources to meet the precise needs of the parents.


Manual of Recommendations for the Diagnosis, Therapy, and Follow-Up of Patients with Breast Cancer of the Tumor Center Munich - a Regional Hands-On Publication.
The revised 11th edition of the Manual of Recommendations for the Diagnosis, Therapy, and Follow-Up of Patients with Breast Cancer of the publications series of the Tumor Center Munich (Tumorzentrum München, TZM) is an excellent example of a regional hands-on publication which, while based on national and international guidelines, does not replace these. By virtue of countless additions and revisions in the course of 10 editions, the 'blue tumor manual for breast cancer' has matured into a hands-on reference work which throughout Germany has found its place on the desks of physicians, and has thus gained a reputation reaching far beyond the TZM. The reputation of this manual is on the one hand founded on the professional competence of the individual authors and project group members. The great strength of the project group, however, surely lies in the broad spectrum of expertise of more than 180 experts of all kinds of specialties, whose continuing interdisciplinary exchange in the course of the present revision has again led to a result which is arousing interest far beyond Munich. This article summarizes some of the TZM project group's own positions on data collection, prevention, adjuvant systemic therapy, and follow-up treatment in an exemplary fashion.


New act expands efforts against fraud and abuse.
President Clinton on August 21 signed into law the Health Insurance Portability and Accountability Act of 1996. Though much of the publicity surrounded passage of the bill centered on the provision of insurance portability, limits on preexisting condition exclusions, and the establishment of a medical savings account pilot project, the bill also includes a major initiative to expand federal efforts to combat fraud and abuse. This overview highlights the most significant elements of that initiative.


Protective effect of intranasal insulin on postoperative cognitive dysfunction in elderly patients with metabolic syndrome undergoing noncardiac surgery: a randomized clinical trial.
Insulin plays a crucial and multifactorial role in cognitive activity, with insulin resistance appearing in neurodegenerative and metabolic diseases. Insulin resistance contributes to the pathobiology of postoperative cognitive dysfunction (POCD) in experimental models, which can be rescued by intranasal insulin administration. To test the effect of intranasal insulin on the incidence of POCD in elderly patients with metabolic syndrome. The study was designed as a randomized, double-blind, placebo-controlled clinical trial. 116 elderly participants were randomly assigned to receive either 40 IU insulin (n = 58) or placebo (n = 58) for 7 days. The primary outcome was the incidence of POCD at 7 days and 3 months after surgery. Secondary outcomes included the degree of peripheral insulin resistance postoperatively, changes in peripheral inflammation levels and the safety of interventions. The results showed that POCD occurred in the insulin group on the 7th postoperative day in 11 (20.8%) patients, which was fewer than the 23 (45.1%) patients in the placebo group (P = 0.008). The insulin group indicated better cognitive functional performance on language and memory test than the placebo group (P < 0.05). Mean peripheral plasma concentration of TNF-α (P < 0.05) and CRP (P < 0.001) in the insulin group was significantly declined compared with the placebo group on D3 and D7. Intranasal insulin administration reduced the incidence of POCD and alleviated peripheral inflammatory levels in elderly patients with metabolic syndrome. Chinese Clinical Trial Registry (ChiCTR1800015502).


Approach and avoidance motivation in psychopathic criminal offenders during passive avoidance.
The authors evaluated competing theories that attribute psychopathic individuals' poor passive avoidance to a strong activating system, a weak inhibitory system, or poor modulation of behavioral activation when inhibitory cues appear. In Study 1, the continuous motor task involved a reward phase to elicit the activating system followed by a passive avoidance phase. Study 2 tested the generality of the theories by using an active avoidance phase to elicit the activating system. Heart rate and response speed results from Study 1 best supported the strong activating system and poor response modulation models in low-anxiety psychopathic offenders. Study 2 results did not clearly support any of the models. Further research is needed to determine if excessive activation by reward and poor response modulation are associated with passive avoidance deficits and other characteristics of low-anxiety psychopathic offenders.


Towards AOP application--implementation of an integrated approach to testing and assessment (IATA) into a pipeline tool for skin sensitization.
Since the OECD published the Adverse Outcome Pathway (AOP) for skin sensitization, many efforts have focused on how to integrate and interpret nonstandard information generated for key events in a manner that can be practically useful for decision making. These types of frameworks are known as Integrated Approaches to Testing and Assessment (IATA). Here we have outlined an IATA for skin sensitization which focuses on existing information including non testing approaches such as QSAR and read-across. The IATA was implemented into a pipeline tool using OASIS technology to provide a means of systematically collating and compiling relevant information which could be used in an assessment of skin sensitization potential. A test set of 100 substances with available skin sensitization information was profiled using the pipeline IATA. In silico and in chemico profiling information alone was able to correctly predict skin sensitization potential, with a preliminary accuracy of 73.85%. Information from other relevant endpoints (e.g., Ames mutagenicity) was found to improve the accuracy (to 87.6%) when coupled with a reaction chemistry mechanistic understanding. This pipeline platform could be useful in the assessment of skin sensitization potential and marks a step change in how non testing approaches can be practically applied.


Evaluation of screening tests for colorectal cancer.
The following guidelines are proposed for the asymptomatic patient representing for routine examination. Instruct the patient to eat All Bran cereal or a similar product for breakfast for three consecutive days prior to the day of appointment. At the time of appointment the stool obtained from rectal examination or from a spontaneous bowel movement is checked for occult blood using the guaiac method. If the findings are negative, no further tests are recommended. If positive, the patient is given complete dietary instructions in a non-meat, high-residue diet with avoidance of beets, horseradish, vitamins, or aspirin-containing compounds. The patient is then given six Hemoccult or Quikcult slides and is instructed to prepare two fecal slides from each stool specimen daily for three days. If these are all negative when tested, no further studies are necessary. If one or more are positive, however, sigmoidoscopic examination and colon and upper gastrointestinal radiography should be carried out in that order. Evidence that early lesions (Duke A or B) are detected and the cure rate improved with this procedure is quite convincing.


Different states of synaptotagmin regulate evoked versus spontaneous release.
The tandem C2-domains of synaptotagmin 1 (syt) function as Ca(2+)-binding modules that trigger exocytosis; in the absence of Ca(2+), syt inhibits spontaneous release. Here, we used proline linkers to constrain and alter the relative orientation of these C2-domains. Short poly-proline helices have a period of three, so large changes in the relative disposition of the C2-domains result from changing the length of the poly-proline linker by a single residue. The length of the linker was varied one residue at a time, revealing a periodicity of three for the ability of the linker mutants to interact with anionic phospholipids and drive evoked synaptic transmission; syt efficiently drove exocytosis when its tandem C2-domains pointed in the same direction. Analysis of spontaneous release revealed a reciprocal relationship between the activation and clamping activities of the linker mutants. Hence, different structural states of syt underlie the control of distinct forms of synaptic transmission.


Mechanism of aggregation of fibrinogen molecules: the influence of fibrin-stabilising factor.
The physicochemical mechanism of aggregation of fibrinogen has been investigated in the presence and absence of fibrin-stabilising factor (factor XIIIa). Data from elastic and inelastic light-scattering and viscometry show that molecules of fibrinogen undergo a spontaneous modification of their carboxyl terminals and bind 'end to end' into flexible polymer chains. On attaining a critical length, the single-filament polymers twist into a coil and aggregate to form branched molecules in which the segments are packed sufficiently densely to resemble strongly hydrated globular particles. The formation, under the influence of factor XIIIa, of epsilon/gamma-glutamyl-lysine covalent bonds produces only insignificant changes in the spatial organisation of the fibrinogen aggregates. Covalent dimerisation of the gamma-chains restricts the structural flexibility of the polymers, but linking of the alpha-chains provides progressive compaction of the structure with increase in molecular weight. Electrophoresis of reconstituted samples shows that the coil-shaped chains of fibrinogen oligomers prevent the complete enzymatic linking of the gamma-chains. The results of this work suggest that the accelerated assembly of multimolecular aggregates, seen in the presence of factor XIIIa, may be explained by the stabilisation of intermediate complexes of fibrinogen, which makes the spontaneous transition from a stable native state to the activated state irreversible.


Mitochondria and cell death.
Mitochondria play a central role in both apoptosis and necrosis through the opening of the mitochondrial permeability transition pore (MPTP). This is thought to be formed through a Ca(2+)-triggered conformational change of the adenine nucleotide translocase (ANT) bound to matrix cyclophilin-D and we have now demonstrated this directly by reconstitution of the pure components. Opening of the MPTP causes swelling and uncoupling of mitochondria which, unrestrained, leads to necrosis. In ischaemia/reperfusion injury of the heart we have shown MPTP opening directly. Recovery of hearts correlates with subsequent closure, and agents that prevent opening or enhance closure protect from injury. Transient MPTP opening may also be involved in apoptosis by initially causing swelling and rupture of the outer membrane to release cytochrome c (cyt c), which then activates the caspase cascade and sets apoptosis in motion. Subsequent MPTP closure allows ATP levels to be maintained, ensuring that cell death remains apoptotic rather than necrotic. Apoptosis in the hippocampus that occurs after a hypoglycaemic or ischaemic insult is triggered by this means. Other apoptotic stimuli such as cytokines or removal of growth factors also involve mitochondrial cyt c release, but here there is controversy over whether the MPTP is involved. In many cases cyt c release is seen without any mitochondrial depolarization, suggesting that the MPTP does not open. Recent data of our own and others have revealed a specific outer-membrane cyt c-release pathway involving porin that does not release other intermembrane proteins such as adenylate kinase. This is opened by pro-apoptotic members of the Bcl-2 family such as BAX and prevented by anti-apoptotic members such as Bcl-X(L). Our own data suggest that this pathway may interact directly with the ANT in the inner membrane at contact sites.


Maternal Embryo Effect Arrest 31 (MEE31) is a moonlighting protein involved in GDP-D-mannose biosynthesis and KAT1 potassium channel regulation.
Due to anthropogenic global warming, droughts are expected to increase and water availability to decrease in the coming decades. For this reason, research is increasingly focused on developing plant varieties and crop cultivars with reduced water consumption. Transpiration occurs through stomatal pores, resulting in water loss. Potassium plays a significant role in stomatal regulation. KAT1 is an inward-rectifying potassium channel that contributes to stomatal opening. Using a yeast high-throughput screening of an Arabidopsis cDNA library, MEE31 was found to physically interact with KAT1. MEE31 was initially identified in a screen for mutants with delayed embryonic development. The gene encodes a conserved phosphomannose isomerase (PMI). We report here that MEE31 interacts with and increases KAT1 activity in yeast and this interaction was also confirmed in plants. In addition, MEE31 complements the function of the yeast homologue, whereas the truncated version recovered in the screening does not, thus uncoupling the enzymatic activity from KAT1 regulation. We show that MEE31 overexpression leads to increased stomatal opening in Arabidopsis transgenic lines. Our data suggest that MEE31 is a moonlighting protein involved in both GDP-D-mannose biosynthesis and KAT1 regulation.


Response of the nitrergic system to activation of the neuroendocrine stress axis.
Exposure to stressful stimuli causes activation of the hypothalamic-pituitary-adrenal axis which rapidly releases high concentrations of glucocorticoid stress hormones, resulting in increased cellular metabolism and spontaneous oxygen and nitrogen radical formation. High concentrations of nitrogen radicals, including nitric oxide, cause damage to cellular proteins in addition to inhibiting components of the mitochondrial transport chain, leading to cellular energy deficiency. During stress exposure, pharmacological inhibition of nitric oxide production reduces indicators of anxiety- and depressive-like behavior in animal models. Therefore, the purpose of this review is to present an overview of the current literature on stress-evoked changes in the nitrergic system, particularly within neural tissue.


[Phasic character of the process of color sensitivity in the visual analyzer].
In subjects with normal three-chromatic vision, during long-term uninterrupted adaptation (6--8 min) to spectral stimuli of different spectral areas (lambda=650+585 nm; lambda=535+585 nm), it was found that the state of color sensitivity undergoes changes in the form of alternating active (correct) and passive (wrong) phases of color discrimination. First, the phase of active color discrimination prevails, then both phases become sort of balanced, and in the end of the adaptation the phase of passive color discrimination increases considerably. The time of duration of these phases depends on the wave--length of the adapting stimulus.


Preventing Alcohol-Exposed Pregnancies: A Randomized Controlled Trial of a Self-Administered Version of Project CHOICES with College Students and Nonstudents.
Alcohol-exposed pregnancies (AEPs) are a preventable cause of birth defects and developmental disabilities for which many women are at risk. The initial 5-session Project CHOICES intervention was found to prevent AEPs. In the ensuing decade, there have been several additional CHOICES-like studies. This study, Project Healthy CHOICES, had 2 objectives: (i) to compare outcomes for students versus nonstudents; and (ii) to test a self-administered mail-based version of the Project CHOICES intervention. A randomized controlled trial (RCT) compared 2 interventions for women of childbearing age (18 to 44) who were at risk of an AEP: (i) motivational feedback based on Project CHOICES and (ii) information only. Advertisements recruited 354 women (145 college students; 209 nonstudents) at risk of an AEP. Intervention and study materials were available in English and Spanish. Of the 354 women, 44% were minorities (25% identified as Hispanics). At the 6-month follow-up, the interventions did not differ and there was no Intervention by Student Study interaction. However, over the entire 6-month follow-up, significantly more students (68%) than nonstudents (46%) were not at risk of an AEP (2.1 odds ratio; confidence interval = 1.47 to 2.95). For all groups, risk reduction occurred primarily through effective contraception. There was no significant difference between the 2 interventions. However, over the entire 6-month follow-up interval, college students were significantly more likely than nonstudents to not be at risk of an AEP and to use effective contraception. While the student groups had significantly higher reduced risk of AEP outcomes, there was also substantial risk reduction for women in the information only condition. These results suggest that the most effective AEP prevention efforts would be to inform women at risk that they could become pregnant. Because about half of all pregnancies are unplanned, identifying women at risk and preventing the risk of AEPs should be a public health priority.


Epidermal growth factor receptor and the adaptor protein p52Shc are specific substrates of T-cell protein tyrosine phosphatase.
T-cell protein tyrosine phosphatase (TCPTP) exists as two forms generated by alternative splicing: a 48-kDa endoplasmic reticulum (ER)-associated form (TC48) and a 45-kDa nuclear form (TC45). To identify TCPTP substrates, we have generated substrate-trapping mutants, in which the invariant catalytic acid of TCPTP (D182) is mutated to alanine. The TCPTP D182A substrate-trapping mutants were transiently overexpressed in COS cells, and their ability to form complexes with tyrosine-phosphorylated (pTyr) proteins was assessed. No pTyr proteins formed complexes with wild-type TCPTP. In contrast, TC48-D182A formed a complex in the ER with pTyr epidermal growth factor receptor (EGFR). In response to EGF, TC45-D182A exited the nucleus and accumulated in the cytoplasm, where it bound pTyr proteins of approximately 50, 57, 64, and 180 kDa. Complex formation was disrupted by vanadate, highlighting the importance of the PTP active site in the interaction and supporting the characterization of these proteins as substrates. Of these TC45 substrates, the approximately 57- and 180-kDa proteins were identified as p52Shc and EGFR, respectively. We examined the effects of TC45 on EGFR signaling and observed that it did not modulate EGF-induced activation of p42Erk2. However, TC45 inhibited the EGF-induced association of p52Shc with Grb2, which was attributed to the ability of the PTP to recognize specifically p52Shc phosphorylated on Y239. These results indicate that TC45 recognizes not only selected substrates in a cellular context but also specific sites within substrates and thus may regulate discrete signaling events.


Small molecular probe as selective tritopic sensor of Al3+, F- and TNP: Fabrication of portable prototype for onsite detection of explosive TNP.
Schiff base organic compound (SOC) has been prepared as a tritopic chemosensor for selective sensing by fluorescence signalling towards ions like Al3+, F- and explosive molecule like TNP. In general, fluorescence like photophysical property has been used for selective detection of analyte where Al3+ and F- show turn-on fluorescence signal at different wavelengths (nm) however, quenching was found with TNP. As a consequence, the chemosensor has become a selective sensor for Al3+, F- and TNP. Reversibility of fluorescence responses for Al3+ and F- are observed in presence of ammonium nitrate and H+ respectively. Similar to the detection of TNP, the detection of explosive like NO3- salts is also essential from homeland security point of view. In the present work, the finding of reversible sequential fluorescence response can be promoted for fabrication of next generation AND-NOT-OR-NAND-XOR-XNOR-NOR based complex logic circuit which is applicable in photonics, security and other fields including inorganic and material science. In the case of TNP recognition, the pathway mainly depends on non-covalent interaction (quenching constant: 4.4 × 105 M-1) which is even better than the recently reported materials. Detection limit for Al3+, F- and TNP is 1 μM, 3 μM and 500 nM respectively. DFT-D3 has been carried out to explore the host⋯guest interaction along with the structure-property correlation of the present host-guest system. All three guest analytes have been detected inside the living cell at a certain level and in its consequence, the successful in vitro recognition ability of the SOCs inside human cell line HeLa has been explored too. In real time stepping, an easy to operate and an economically affordable pocket prototype has also been fabricated for on spot detection of TNP like explosive.


Color mismatch and observer metamerism between conventional liquid crystal displays and organic light emitting diode displays.
Organic light emitting diode (OLED) displays use red, green, and blue primaries with a higher saturation level to produce larger color gamuts than conventional liquid crystal displays (LCD). No past study, however, experimentally investigated how such a difference between these two display types causes color mismatch and observer metamerism using the most widely used color matching functions (CMFs)-the CIE 1931 2° CMFs-for color calibration and specification. In this study, 50 human observers performed color matching tasks for six color stimuli with a field-of-view of 4.77° between four test displays (i.e., one LCD and three OLED) and a reference OLED display. The color gamuts of the LCD and OLED displays were similar to the sRGB and P3 standard color gamuts. It was found the CIE 1931 2° CMFs cannot accurately characterize the color matches between the LCD and OLED displays, with different chromaticities required to produce matched color appearance. Particularly, when the stimuli had matched color appearance, the chromaticities of the stimuli produced by the LCD display were all shifted towards the -u'+v' direction in the CIE 1976 u'v' chromaticity diagram in comparison to those produced by the OLED display. This suggested that using the CIE 1931 2° CMFs for display calibration would cause the colors shown on OLED displays to have a yellow-green tint if those on LCD displays appear neutral. In addition, a larger degree of observer metamerism was found between the LCD and OLED displays, while little differences, in terms of color mismatch and observer metamerism, were found between the OLED displays. The CIE 2006 2° CMFs were found to have better performance than the CIE 1931 2°, 1964 10°, and 2006 10° CMFs, which could be partially due to the size of the stimulus used in the experiment.


An evaluation for compensated anterior cavus.
Anterior cavus is a multiplane structural deformity that can originate from different areas within the foot. Differentiation of these areas is a necessary examination in order to determine the effect of proper treatment. A depressed fourth and fifth ray or lateral column is a major deformity in anterior cavus. The lateral column deformity is a pronatory force that causes a premature medial shift of weight. The premature shift can cause a breakdown of the medial column of the foot. We have developed an examination of the compensated anterior cavus deformity that enables the practitioner to isolate better the pronatory forces within the foot.


NeuroD/BETA2 gene variability and diabetes: no associations to late-onset type 2 diabetes but an A45 allele may represent a susceptibility marker for type 1 diabetes among Danes. Danish Study Group of Diabetes in Childhood, and the Danish IDDM Epidemiology and Genetics Group.
Mutations in the NeuroD/BETA2 gene have been shown to associate with type 2 diabetes. In the present study, we examined mutations in the NeuroD/BETA2 gene for association with either type 1 or 2 diabetes. Three variants were identified in patients with type 2 diabetes: Ala45Thr (allelic frequency 0.36, 95% CI 0.31-0.41), Pro197His (0.01), and Ser259Ser (0.01). Ala45Thr and Pro197His were not associated with type 2 diabetes, but the transmission disequilibrium test showed unequal transmission of the A45 allele to offspring with type 1 diabetes (chi2 = 5.90, P < 0.02, odds ratio 1.55, 95% CI 0.91-2.63). This association could not be explained by linkage disequilibrium between the Ala45 allele and IDDM7 (D2S152), which is also located on chromosome 2q32. When tested in vitro, the biological activity of Thr45 (117+/-36% vs. Ala45) and His197 (90+/-28% vs. Pro197) on the regulation of the human insulin gene promoter appeared normal. In conclusion, mutations in the NeuroD/BETA2 gene are not a common cause of late-onset type 2 diabetes among Danes. However, in the type 1 diabetic Danish population, the Ala45Thr variant of NeuroD/BETA2 may represent a susceptibility marker independent of IDDM7 on chromosome 2q32.


Improving Adherence to Myofunctional Therapy in the Treatment of Sleep-Disordered Breathing.
Myofunctional therapy (MT) is used to treat sleep-disordered breathing. However, MT has low adherence-only ~10% in most studies. We describe our experiences with MT delivered through a mobile health app named Airway Gym®, which is used by patients who have rejected continuous positive airway pressure and other therapies. We compared ear, nose, and throat examination findings, Friedman stage, tongue-tie presence, tongue strength measured using the Iowa oral performance instrument (IOPI), and full polysomnography before and after the 3 months of therapy. Participants were taught how to perform the exercises using the app at the start. Telemedicine allowed physicians to record adherence to and accuracy of the exercise performance. Fifty-four patients were enrolled; 35 (64.8%) were adherent and performed exercises for 15 min/day on five days/week. We found significant changes (p < 0.05) in the apnoea-hypopnoea index (AHI; 32.97 ± 1.8 to 21.9 ± 14.5 events/h); IOPI score (44.4 ± 11.08 to 49.66 ± 10.2); and minimum O2 saturation (80.91% ± 6.1% to 85.09% ± 5.3%). IOPI scores correlated significantly with AHI after the therapy (Pearson r = 0.4; p = 0.01). The 19 patients who did not adhere to the protocol showed no changes. MT based on telemedicine had good adherence, and its effect on AHI correlated with IOPI and improvement in tongue-tie.


A very unusual case of food allergy, between FPIES and IgE-mediated food allergy.
Food protein induced enterocolitis syndrome (FPIES) is a food-related gastrointestinal hypersensitivity disorder, probably non-IgE-mediated. Over the years, various diagnostic criteria have been proposed to identify FPIES. In the last few years, there was an increased interest from researchers about FPIES's syndrome, that frequently brought to discover new aspects of this disease. We describe an unusual case of FPIES to egg in a 21-months-old child, because of its clinical characteristics that reflect some aspects of IgE-mediated allergy and other of non IgE-mediated allergy. Although we believe that the most correct diagnosis for our case is FPIES, we think also that this is undoubtedly an atypical form. This is in fact, the first description of a patient who simultaneously has both clinical expressions of IgE-mediated FA that of FPIES. Our case highlights the need to review criteria for FPIES diagnosis.


Severe Postoperative Complications may be Related to Mesenteric Traction Syndrome during Open Esophagectomy.
During abdominal surgery, traction of the mesenterium provokes mesenteric traction syndrome, including hypotension, tachycardia, and flushing, along with an increase in plasma prostacyclin (PGI2). We evaluated whether postoperative complications are related to mesenteric traction syndrome during esophagectomy. Flushing, hemodynamic variables, and plasma 6-keto-PGF1α were recorded during the abdominal part of open ( n = 25) and robotically assisted ( n = 25) esophagectomy. Postoperative complications were also registered, according to the Clavien-Dindo classification. Flushing appeared in 17 (open) and 5 (robotically assisted) surgical cases ( p = 0.001). Mean arterial pressure was stable during both types of surgeries, but infusion of vasopressors during the first hour of open surgery was related to development of widespread (Grade II) flushing ( p = 0.036). For patients who developed flushing, heart rate and plasma 6-keto-PGF1α also increased ( p = 0.001 and p < 0.001, respectively). Furthermore, severe postoperative complications were related to Grade II flushing ( p = 0.037). Mesenteric traction syndrome manifests more frequently during open than robotically assisted esophagectomy, and postoperative complications appear to be associated with severe mesenteric traction syndrome.


Design and Photonics of Merocyanine Dyes.
Merocyanines, thanks to their easily adjustable electronic structure, appear to be the most versatile and promising functional dyes. Their D-π-A framework offers ample opportunities for custom design through variations in both donor/acceptor end-groups and the π-conjugated polymethine chain, and leads to a broad range of practical properties, including noticeable solvatochromism, high polarizability/hyperpolarizabilities, and the ability to sensitize various physicochemical processes. Accordingly, merocyanines are applied and extensively studied in various fields, such as light-converting materials for optoelectronics, nonlinear optics, optical storage, solar cells, fluorescent probes, and antitumor agents in photodynamic therapy. This review encompasses both classical and novel more important publications on the structure-property relationships in merocyanines, with particular emphasis on the results by A. I. Kiprianov and his followers in Institute of Organic Chemistry in Kyiv, Ukraine.


Substance P in the dorsal vagal complex inhibits medullary TRH-induced gastric acid secretion in rats.
Neurons that contain substance P (SP) and thyrotropin-releasing hormone (TRH) in medullary midline raphe nuclei project to the dorsal vagal complex (DVC). The modulatory role of SP on basal gastric acid secretion (GAS) and TRH on DVC-induced stimulation of GAS was studied in urethan-anesthetized rats. The stable SP agonist, DiMe-C7 ([pGlu5, MePhe8, MeGly9]SP5-11, 50 and 100 pmol), injected unilaterally into the DVC reduced the GAS response (47 +/- 12 mumol/60 min) to coinjected TRH analog, RX 77368 (25 pmol), by 53% and 85%, respectively, whereas DiMe-C7 (100 pmol) alone had no effect on basal and pentagastrin-stimulated GAS. DiMe-C7 (100 pmol/site) inhibited the GAS response to kainic acid injected into the raphe pallidus (Rpa) when it was injected bilaterally into the DVC but not the hypoglossal nuclei. The SP nourokinin-1-receptor antagonist, CP-96,345, injected bilaterally into the DVC (1 nmol/ site) increased basal GAS (33 +/- 8 mumol/90 min) and potentiated the GAS response to kainic acid injected into the Rpa by 40%. These results suggest that SP acts on neurokinin-1 receptors in the DVC to reduce medullary TRH-induced stimulation of GAS in rats.


Analysis of a claimed distant relationship in a deficient pedigree using high density SNP data.
DNA markers are routinely used to reveal both simple and complex family relationships. Likelihood based approaches have been traditionally used to estimate relationships using relatively few unlinked markers. However it is widely recognized that when using such limited numbers of loci distant relationships between two individuals cannot be distinguished from the average level of allele sharing found in random pairwise comparisons in the same population. As a real example, we demonstrate the usefulness of genome-wide SNP genotyping to analyze a claimed second cousin relationship that could not be resolved using standard forensic markers, confirming theoretical expectations for very distant relationships. Genome profiles derived from Affymetrix 6.0 SNP arrays obtained from the claimed second cousins were compared to profiles obtained from unrelated individuals and simulated data. Significance of the high estimated probabilities in favor of the second cousin relationship hypothesis was proved from the results obtained with both real and simulated unrelated pairs. As a final cautionary note, it is important to consider that successful identification of the claimed distant relationship reported here is largely due to a well-founded hypothesis being compared to the alternative hypothesis of the claimants being unrelated, but where there are several possible alternative hypotheses, the approach we outline here can yield false indications of unfounded alternative relationships.


Infants' Transitions out of a Fussing/Crying State Are Modifiable and Are Related to Weight Status.
Currently, about 10% of infants have a weight for length greater than the 95th percentile for their age and sex, which puts them at risk for obesity as they grow. In a pilot obesity prevention study, primiparous mothers and their newborn infants were randomly assigned to a control group or a Soothe/Sleep intervention. Previously, it has been demonstrated that this intervention contributed to lower weight-for-length percentiles at 1 year; the aim of the present study was to examine infant behavior diary data collected during the intervention. Markov modeling was used to characterize infants' patterns of behavioral transitions at ages 3 and 16 weeks. Results showed that heavier mothers were more likely to follow their infants' fussing/crying episodes with a feeding. The intervention increased infants' likelihood of transitioning from a fussing/crying state to an awake/calm state. A shorter latency to feed in response to fussing/crying was associated with a higher subsequent weight status. This study provides preliminary evidence that infants' transitions out of fussing/crying are characterized by inter-individual differences, are modifiable, and are linked to weight outcomes, suggesting that they may be promising targets for early behavioral obesity interventions, and highlighting the methodology used in this study as an appropriate and innovative tool to assess the impact of such interventions.


[Single centre experience of endoscopic submucosal dissection (ESD) in premalignant and malignant gastrointestinal neoplasia].
Worldwide endoscopic submucosal dissection (ESD) of early GI cancers or premalignant neoplasia is becoming increasingly important. In Germany ESD is restricted to larger endoscopic institutions and only a few literature reports are available. The aim of the present study is to describe the results of 46 ESDs conducted in a German endoscopic centre. Between June 2007 and May 2012 46 ESDs in 45 patients (33 men, 12 women, mean age 66.1 years) were performed in the oesophagus (n = 17), stomach (n = 23) and rectum (n = 6). Data were collected for the en-bloc, R0 and R0 en-bloc resection rates as well as for complications, the curative resection and the local recurrence rates. In order to demonstrate a learning curve, results were evaluated for two periods (June 2007 to November 2010 vs. December 2010 to May 2012). ESD was technically possible in 93.5%. En-bloc, R0, R0 en-bloc and curative resection rates were 90.7%, 74.4%, 67.4% and 65.1%, respectively. The complication rate was 13%. In the second period en-bloc and R0 en-bloc resection rates increased from 81% to 100% and, respectively, from 52.4% to 81.8%. After a medium follow-up of 11.4 months, local tumour recurrence occurred in 10%. In cases of curative R0 en bloc resection of malignant tumours no tumour recurrence occurred. Despite the small number of patients, the present data underline the value of ESD, especially in cases of R0 en-bloc resections in the therapy for premalignant and early malignant GI tumours. Due to the required learning curve, ESD should be restricted to larger endoscopic centres in Germany.


Biological properties of polyoma DNA fragments cloned in plasmid pBR322.
The two HindIII fragments of polyoma virus DNA were cloned in the HindIII site of plasmid pBR322, and the biological activity of the recombinant plasmids was tested in tissue culture cells. A mixture of recombinant plasmids containing the HindIII-A and HindIII-B fragments was infectious, but only after cleavage with HindIII. Recombinant plasmids containing the HindIII-A fragment, but not those containing the HindIII-B fragment, induced the transformation of Fischer rat 3T3 cells. These findings indicate that about half of the early region of polyoma virus DNA is not essential for the initiation of the maintenance of transformation.


SPATIAL UPDATING OF HAPTIC ARRAYS ACROSS THE LIFE SPAN.
Background/Study Context: Aging research addressing spatial learning, representation, and action is almost exclusively based on vision as the input source. Much less is known about how spatial abilities from nonvisual inputs, particularly from haptic information, may change during life-span spatial development. This research studied whether learning and updating of haptic target configurations differs as a function of age. Three groups of participants, ranging from 20 to 80 years old, felt four-target table-top circular arrays and then performed several tasks to assess life-span haptic spatial cognition. Measures evaluated included egocentric pointing, allocentric pointing, and array reconstruction after physical or imagined spatial updating. All measures revealed reliable differences between the oldest and youngest participant groups. The age effect for egocentric pointing contrasts with previous findings showing preserved egocentric spatial abilities. Error performance on allocentric pointing and map reconstruction tasks showing a clear age effect, with the oldest participants exhibiting the greatest error, is in line with other studies in the visual domain. Postupdating performance sharply declined with age but did not reliably differ between physical and imagined updating. Results suggest that there is a general trend for age-related degradation of spatial abilities after haptic learning, with the greatest declines manifesting in all measures in people over 60 years of age. Results are interpreted in terms of a spatial aging effect on mental transformations of three-dimensional representations of space in working memory.


Angiomyolipoma of the colon: report of a case and review of the literature.
Angiomyolipoma of the colon is very rare. Only three cases have been reported in the literature. Here we report the case of a 54-year-old male, who presented with a progressive abdominal cramping pain. We performed left hemicolectomy under the impression of a cancerous mass over the splenic flexure of the colon. Histology revealed an angiomyolipoma of the colon. In addition, we review the literature.


Neonatal Marfan syndrome: Report of two cases.
Marfan syndrome is rarely diagnosed in the neonatal period because of variable expression and age-dependent appearance of clinical signs. The prognosis is usually poor due to high probability of congestive heart failure, mitral and tricuspid regurgitations with suboptimal response to medical therapy and difficulties in surgical management. The authors have studied two cases of Marfan syndrome in the newborn period. Two cases of neonatal Marfan syndrome, one male and one female, were diagnosed by characteristic physical appearance. Both infants had significant cardiovascular abnormalities diagnosed by ultrasonography. Genetic DNA analysis in the second case confirmed the mutations in the fibrillin-1 gene located on chromosome 15q21 which is responsible for the development of Marfan syndrome. The boy died at six weeks of age with signs of rapidly progressive left ventricular failure associated with pneumonia. The second infant was having only mild signs of congestive heart failure and has been treated with beta blockers. At the age of 4 years her symptoms of congestive heart failure had worsened due to progression of mitral and tricuspid insufficiency and development of significant cardiomegaly. Mitral and tricuspid valvuloplasy had to be done at that time. Early diagnosis of Marfan syndrome in the newborn period can allow treatment in the early stages of cardiovascular abnormalities and may improve the prognosis. It also helps to explain to the family the serious health problem of their child.


Normative distribution of visual acuity in 3- to 6-year-old Chinese preschoolers: the Shenzhen kindergarten eye study.
To document the distribution of uncorrected visual acuity (UCVA) in a defined population of Chinese preschoolers and to discuss its implications for vision referral. Preschoolers aged 3 to 6 years old were recruited from kindergartens in Shenzen. Uncorrected visual acuity was estimated by using Early Treatment Diabetic Retinopathy Study Tumbling E charts, followed by cycloplegic refraction and ocular examination. The reference population was defined as children without clinically significant ocular abnormalities, with spherical equivalent refraction greater than -0.50 to less than +2.00 diopters (D), astigmatism less than 0.75 D, and anisometropia less than 2.00 D. The age-specific UCVA cutoffs were defined by the line where the single-sided 95th percentile of the reference population fell. A total of 483 of the 1255 children enrolled were considered the reference population. The monocular UCVA cutoff fell on the line of 20/63 at age 3, 20/50 at age 4, and 20/40 at ages 5 and 6. Using no better than these lines as criteria generated referral rates of 9.4% to 27.8% in the general population at different ages, and detected 83.3% and more than 90.0% of those with myopia and amblyopia, respectively. Using uncorrected interocular difference of two or more lines referred 3.6% to 4.3% of the population but identified only approximately 20.0% of those with amblyopia. Visual acuity is still developing in preschoolers even at age 6. Most children with myopia and amblyopia can be identified with age-specific, monocular UCVA cutoffs in vision screening using Tumbling E charts, with tolerable false-positive rates. Further studies are needed to define the age at which children without significant refractive errors reach 20/20 UCVA.


Liquid tissue surrogates for X-ray and CT phantom studies.
To develop a simple method for producing liquid-tissue-surrogate (LTS) materials that accurately represent human soft tissues in terms of density and X-ray attenuation coefficient. We evaluated hypothetical mixtures of water, glycerol, butanol, methanol, sodium chloride, and potassium nitrate; these mixtures were intended to emulate human adipose, blood, brain, kidney, liver, muscle, pancreas, and skin. We compared the hypothetical densities, effective atomic numbers (Zeff ), and calculated discrete-energy CT attenuation [Hounsfield Units (HU)] of the proposed materials with those of human tissue elemental composition as specified in International Commission on Radiation Units (ICRU) Report 46. We then physically produced the proposed LTS materials for adipose, liver, and pancreas tissue, and we measured the polyenergetic CT attenuation (also expressed as HU) of these materials within a 32 cm phantom using a 64-slice clinical CT scanner at 80 kVp, 100 kVp, 120 kVp, and 140 kVp. The predicted densities, Zeff , and calculated discrete-energy CT attenuation of our proposed formulations generally agreed with those of ICRU within < 1% or < 10 HU. For example, the densities of our hypothetical materials agreed precisely with ICRU's reported values and were 0.95 g/mL for adipose tissue, 1.04 g/mL for pancreatic tissue, and 1.06 g/mL for liver tissue; the discrete-energy CT attenuation at 60 keV of our hypothetical materials (and ICRU-specified compositions) were -107 HU (-113 HU) for adipose #3, -89 HU (-90 HU) for adipose #2, 56 HU (55 HU) for liver tissue, and 31 HU (31 HU) for pancreatic tissue. The densities of our physically produced materials (compared to ICRU-specified compositions) were 0.947 g/mL (0.0%) for adipose #2, 1.061 g/mL (+2.0%) for pancreatic tissue, and 1.074 g/mL (+1.3%) for liver tissue. The empirical polyenergetic CT attenuation measurements of our LTS materials (and the discrete-energy HU of the ICRU compositions at the mean energy of each spectrum) at 80 kVp were -104 HU (-113 HU) for adipose #3, -87 HU (-90 HU) for adipose #2, 59 HU (55 HU) for liver tissue, and 33 HU (31 HU) for pancreatic tissue; at 120 kVp, these were -83 HU (-83 HU) for adipose #3, -68 HU (-63 HU) for adipose #2, 55 HU (52 HU) for liver tissue, and 35 HU (33 HU) for pancreatic tissue. Our method for formulating tissue surrogates allowed straightforward production of solutions with CT attenuation that closely matched the target tissues' expected CT attenuation values and trends with kVp. The LTSs' inexpensive and widely available constituent chemicals, combined with their liquid state, should enable rapid production and versatile use among different phantom and experiment types. Further study is warranted, such as the inclusion of contrast agents. These liquid tissue surrogates may potentially accelerate development and testing of advanced CT imaging techniques and technologies.


Wearable knee health system employing novel physiological biomarkers.
Knee injuries and chronic disorders, such as arthritis, affect millions of Americans, leading to missed workdays and reduced quality of life. Currently, after an initial diagnosis, there are few quantitative technologies available to provide sensitive subclinical feedback to patients regarding improvements or setbacks to their knee health status; instead, most assessments are qualitative, relying on patient-reported symptoms, performance during functional tests, and physical examinations. Recent advances have been made with wearable technologies for assessing the health status of the knee (and potentially other joints) with the goal of facilitating personalized rehabilitation of injuries and care for chronic conditions. This review describes our progress in developing wearable sensing technologies that enable quantitative physiological measurements and interpretation of knee health status. Our sensing system enables longitudinal quantitative measurements of knee sounds, swelling, and activity context during clinical and field situations. Importantly, we leverage machine-learning algorithms to fuse the low-level signal and feature data of the measured time series waveforms into higher level metrics of joint health. This paper summarizes the engineering validation, baseline physiological experiments, and human subject studies-both cross-sectional and longitudinal-that demonstrate the efficacy of using such systems for robust knee joint health assessment. We envision our sensor system complementing and advancing present-day practices to reduce joint reinjury risk, to optimize rehabilitation recovery time for a quicker return to activity, and to reduce health care costs.


Presence of mecA-positive multidrug-resistant Staphylococcus epidermidis in bovine milk samples in Brazil.
Bacteria of the genus Staphylococcus are one of the major pathogens causing bovine mastitis. In recent decades, resistance of this genus to oxacillin (methicillin) has been a matter of concern due to the possibility of reducing the effectiveness of mastitis treatments and the transfer of resistance determinants to other bacteria. Oxacillin resistance was studied in 170 staphylococci from bovine milk samples, including 79 Staphylococcus aureus and 91 coagulase-negative staphylococci (CNS). The susceptibility profile of 10 antimicrobial agents used in veterinary practice was determined by the Etest method. In addition to the Etest, the phenotypic characterization of oxacillin resistance was tested using the cefoxitin disk diffusion test. All isolates were screened by PCR to detect the mecA gene in 2 different regions of the gene. The isolates with an oxacillin minimum inhibitory concentration ≥0.5 µg/mL or resistant to cefoxitin were identified by sequencing a 536-bp fragment of the 16S rRNA gene. This group of isolates was also evaluated for the presence of blaZ and mecC genes. Molecular analysis of the mecA gene was carried out by typing of the staphylococcal cassette chromosome mec (SCCmec). The relatedness of the mecA-positive isolates was evaluated by macrorestriction of chromosomal DNA followed by pulsed-field gel electrophoresis. With the exception of penicillin and oxacillin, 86% of the isolates showed susceptibility to cephalothin, gentamicin, erythromycin, sulfonamide, trimethoprim-sulfamethoxazole, and tetracycline. All S. aureus isolates were susceptible to oxacillin, whereas 47% (n=43) of the CNS isolates were resistant. The CNS isolates showed a higher resistance to cephalothin, erythromycin, tetracycline, and gentamicin in comparison with S. aureus. The mecA gene was only detected in 10 CNS isolates, identified as Staphylococcus epidermidis, and classified into 3 pulsotypes (A, B, and C) and 4 subtypes (A1, B1, B2, and B3). Among the isolates with an oxacillin resistance phenotype, 12 were positive for the blaZ gene, and 9 of them were mecA-positive. Two of the oxacillin-resistant isolates amplified the mecA homolog gene of Staphylococcus sciuri and none amplified mecC. Three SCCmec types, I, IV, and V, were found. Our results suggest that Staphylococcus epidermidis can be a reservoir for mecA for other Staphylococcus species. Studies investigating the molecular and phenotypic profile of antimicrobial resistance in staphylococcal species should be performed for controlling the spread of resistance and the selection of appropriate therapeutic measures.


APOE and CYP2E1 polymorphisms, alcohol consumption, and Parkinson's disease in a Japanese population.
Apolipoprotein E (APOE) is associated with increased oxidative stress, which is caused by reactive oxygen species (ROS). Enhanced cytochrome P450 2E1 (CYP2E1) activity may also increase formation of neurotoxins such as ROS. As Parkinson's disease (PD) is a neurodegenerative disorder, both the APOE and CYP2E1 genes that are involved in neurodegeneration by oxidative stress may be associated with PD risk. We investigated the relationship of the APOE and CYP2E1 rs2864987 polymorphisms and PD risk with special attention to the interaction with alcohol consumption among 238 patients with PD and 296 controls in a Japanese population. The frequencies of the ɛ2, ɛ3, and ɛ4 alleles of the APOE polymorphism among controls were 3.72, 86.7, and 9.63%, respectively. As compared with the APOE ε3/ε3 genotype, the 2/ε4 genotype was associated with an increased risk of PD (adjusted odds ratio (OR) = 9.50, 95% (confidence interval) CI = 1.12-80.6). The presence of the ε3 allele was associated with a decreased risk of PD. Meanwhile, CYP2E1 rs2864987 was not associated with PD risk. Although CYP2E1 is involved in the metabolism of alcohol, there was no evidence of interaction between alcohol consumption and CYP2E1 rs2864987. Our results suggested that the APOE polymorphism might play an important role in PD susceptibility in our Japanese population. Future studies involving larger control and case populations and better alcohol consumption histories will undoubtedly lead to a more thorough understanding of the role of polymorphisms of genes related to the generation of ROS in PD development.