Gene replacements and insertions in rice by intron targeting using CRISPR-Cas9.
Sequence-specific nucleases have been exploited to create targeted gene knockouts in various plants(1), but replacing a fragment and even obtaining gene insertions at specific loci in plant genomes remain a serious challenge. Here, we report efficient intron-mediated site-specific gene replacement and insertion approaches that generate mutations using the non-homologous end joining (NHEJ) pathway using the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) system. Using a pair of single guide RNAs (sgRNAs) targeting adjacent introns and a donor DNA template including the same pair of sgRNA sites, we achieved gene replacements in the rice endogenous gene 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) at a frequency of 2.0%. We also obtained targeted gene insertions at a frequency of 2.2% using a sgRNA targeting one intron and a donor DNA template including the same sgRNA site. Rice plants harbouring the OsEPSPS gene with the intended substitutions were glyphosate-resistant. Furthermore, the site-specific gene replacements and insertions were faithfully transmitted to the next generation. These newly developed approaches can be generally used to replace targeted gene fragments and to insert exogenous DNA sequences into specific genomic sites in rice and other plants.


[Rhinoscleroma in an immigrant].
A patient with rhinoscleroma was treated in our department. As far as we know it is the first importcase published in Belgium. Rhinoscleroma is an infectious disease caused by the Klebsiella Rhinoscleromatising. It is a chronic granulomatous process with subsequent stenosis and scarring. The nose and larynx are the most frequent localisations. The therapy consists of antibiotics in an early stage and is only palliative in the latter ones.


Pattern of accumulation of elastin and the level of mRNA for elastin in aortic tissue of growing chickens.
Synthesis and accumulation of elastin in many elastic tissues begins in the last third of fetal development, reaches a maximum shortly after birth, and then declines rapidly. For the aorta of the chick and the pig and the ligamentum nuchae and lung of the sheep, it has been shown that increased levels of elastin production with fetal development are correlated with increased levels of elastin mRNA in the tissue, measured both by cell-free translation and by hybridization to cDNA probes. In this study we examine the relationship between insoluble elastin accumulation and message levels for tropoelastin in aortic tissue of chickens during posthatching development and growth. Whether evaluated by cell-free translation or by dot blot hybridization, steady state levels of tropoelastin message increase to a maximum at 2 weeks after hatching, and then fall rapidly with further development and growth. This pattern correlates well with production of insoluble elastin by the aorta, determined either by direct measurements of synthesis or by rate of accumulation of insoluble elastin. The data indicate that the major site of regulation of elastin production is pretranslational throughout the entire period of development and growth of the chicken aorta.


Formaldehyde: an analysis of its respiratory, cutaneous, and immunologic effects.
Formaldehyde is truly ubiquitous in our ecology and continuing important commercial applications. Most of us have daily contact with this chemical. The most significant outdoor source of this chemical is gasoline and diesel fuel combustion. The primary indoor source is combustion of tobacco products. Formaldehyde is associated with a disagreeable odor that can produce "annoyance" symptoms and at higher concentrations can be a transient and completely reversible irritant to the eyes and mucous membranes of the respiratory tract. It is so soluble and rapidly metabolized that it rarely reaches the lower respiratory tract to inflict damage. The exception is in cigarette smokers who actively inhale. Formaldehyde may on rare occasions induce bronchial asthma at relatively high exposure doses. There are no conclusive studies that prove the development of de novo IgE-mediated respiratory tract symptoms secondary to inhalation of formaldehyde vapors. The approach to formaldehyde-induced symptoms should be one of careful documentation of objective physiologic changes.


How a high working memory capacity can increase proactive interference.
Previous findings suggested that a high working memory capacity (WMC) is potentially associated with a higher susceptibility to proactive interference (PI) if the latter is measured under high cognitive load. To explain such a finding, we propose to consider susceptibility to PI as a net effect of individual executive processes and the intrinsic potential for PI. With the latter, we refer to the amount of information that is activated at a given time and that has the potential to exert PI subsequently. In two studies deploying generalized linear mixed models, susceptibility to PI was modeled as the decline of performance over trials of a complex span task. The results revealed that a higher WMC was associated with a higher susceptibility to PI. Moreover, the number of stimuli recalled in one trial as a proxy variable for the intrinsic potential for PI negatively affected memory performance in the subsequent trial.


Use of endobronchial ultrasound to evaluate nonthrombotic endovascular lesions in pulmonary arteries: a systematic review.
The finding of a filling defect in a pulmonary artery (PA) sometimes raises the possibility of cancer. Endobronchial ultrasound (EBUS)-guided transbronchial needle aspiration (TBNA) may confirm the underlying nature of the endovascular lesion. However, little is known about the use of this procedure for this purpose. We searched PubMed and abstracts of major international conferences. There was a total of 12 cases: 8 female, 3 male, and 1 unknown. The median (range) age was 60 (51 to 79) years. EBUS was performed to evaluate mass-like lesion involving the PA (n=5), persistent or progressive filling defects in the PA despite anticoagulation (n=3), filling defect in the PA with multiple areas of consolidation, air-fluid levels in the lung (n=1), and hilar density (n=1). Moreover, an endovascular lesion was incidentally noted in the PA during EBUS for evaluating lymph nodes (n=2). EBUS-TBNA was diagnostic in 9 of the 10 cases in which it was performed. The final diagnoses were: sarcoma (n=6), lung cancer (n=2), thyroid cancer (n=1), renal cell cancer (n=1), melanoma (n=1), and pulmonary embolism (n=1). The cancer was a recurrence in 6 of the 7 cases with a known history of cancer. EBUS should be considered as a possible method for evaluating endovascular lesions when PA sarcoma or tumor macroembolism is suspected.


Clinical experiences with laser-welded titanium frameworks supported by implants in the edentulous mandible: a 10-year follow-up study.
Long-term follow-up studies for more than 5 years are not available on laser-welded titanium frameworks. To report and compare 10-year data on implant-supported prostheses in the edentulous mandible provided with laser-welded titanium frameworks and conventional gold alloy frameworks. Altogether, 155 patients were consecutively treated with prostheses at abutment level with two generations of fixed laser-welded titanium frameworks (test groups). A control group of 53 randomly selected patients with conventional gold alloy castings was used for comparison. Clinical and radiographic 10-year data were collected for the three groups. All patients followed-up for 10 years (n=112) still had fixed prostheses in the mandible (cumulative success rate [CSR] 100%). The overall 10-year cumulative success rate (CSR) was 92.8 and 100.0% for titanium and gold alloy frameworks, respectively. Ten-year implant cumulative survival rate (CSR) was 99.4 and 99.6% for the test and control groups, respectively. Average 10-year bone loss was 0.56 (SD 0.45) mm for the titanium group and 0.77 (SD 0.36) mm for the control group (p < 0.05). The most common complications for titanium frameworks were resin or veneer fractures, soft tissue inflammation, and fractures (12.9%) of the metal frame. Loose and fractured implant screw components were below 3%. Excellent overall long-term results with 100% CSR could be achieved with the present treatment modality. Fractures of the metal frames and remade prostheses were more common for the laser-welded titanium frameworks, and the first generation of titanium frameworks worked poorly when compared with gold alloy frameworks during 10 years (p < 0.05). However, on average more bone loss was observed for implants supporting gold alloy frameworks during 10 years. The reasons for this difference are not clear.


Verteporfin: a review of its use in the management of subfoveal choroidal neovascularisation.
Photodynamic therapy (PDT) with verteporfin (Visudyne), a photosensitising protoporphyrin derivative, is used in the management of subfoveal choroidal neovascularisation (CNV) secondary to age-related macular degeneration (AMD) or pathological myopia (PM). PDT with verteporfin over 1 and 2 years reduces the decline in visual acuity in patients with classic-containing subfoveal CNV secondary to AMD. Verteporfin is generally well tolerated by most patients. Verteporfin is also effective in patients with CNV secondary to PM, although data in this indication are limited and further controlled studies are required. Although verteporfin has shown efficacy in patients with occult AMD-related subfoveal CNV lesions in early trials, data are currently limited on its first-line use in this indication; fully published data from the Verteporfin In Occult (VIO) trial are therefore awaited with interest. Verteporfin should be considered as a first-line treatment in patients with predominantly classic subfoveal CNV secondary to AMD, and in patients with smaller minimally classic subfoveal CNV lesions. It may also be considered an option for the treatment of patients with occult AMD-related subfoveal CNV in whom visual acuity decreases or predominantly classic features develop over time.


Infrainguinal CTO Recanalization Assessed by Intravascular Ultrasound: Results of the CENTRAL Study.
CENTRAL was a prospective, multicenter, 100 patient study designed to evaluate the ability of a recanalization catheter system to cross chronic total occlusions (CTOs) of the superficial femoral artery while staying within the central vessel lumen. The primary endpoint was the successful crossing of a CTO (≥90% of the length) with the recanalization catheter in the central lumen of the superficial femoral artery in at least one-half of the studied patient population, confirmed by intravascular ultrasound. Secondary endpoints included standardized anatomic damage assessment (the TAPE method) and an assessment of the relationship between the percent of CTO crossing and TAPE scores to the rate of target-lesion revascularization (TLR) at 30 days and 6 months. The mean age of the occlusions was 16.6 ± 22.28 months (range, 1-120 months), the average occlusion length reported by the sites was 132.1 ± 87.69 mm (range, 4-300 mm), and a majority of lesions were moderately (42.0%) or severely (32.0%) calcified. In 43/85 (50.6%) of the evaluable intravascular ultrasound images, the recanalization catheter successfully navigated the central lumen of the CTO (ie, ≥90% luminal crossing) with >50% luminal crossing in 64/85 (75.3%) of cases. The 6-month TLR rate was significantly lower in patients where the CTO was crossed ≥90% in the central vessel lumen (4.7%) compared with crossings <90% (20.6%; P=.04). The 6-month TLR rate was 3.5% in patients with a favorable TAPE score of 0-4 and 36.8% with an unfavorable TAPE score of 5-8 (P<.001). Use of a recanalization catheter in complex superficial femoral artery CTO crossings achieved intraluminal crossings in >50% of cases and decreased anatomic damage, which appeared to offer a significant advantage in 6-month TLR rates.


Whole-genome phylogenetic analysis of herpesviruses.
A comprehensive phylogenetic analysis of 45 herpesviruses was performed based on whole-genome sequences. We used 4 methods, namely the alignment of conserved gene sequences (excluding 5 herpesviruses), compositional vector tree (CVTree) method, local homology analysis, and gene content analysis. The obtained results showed good consistency between the phylogenetic trees prepared by these methods and likewise, the obtained classification of the herpesviruses was consistent with their current taxonomic designation. The herpesviruses with the ambiguous classification or not assigned in the family or with the newly published genomes were also phylogenetically classified.


Long-term results of living donor liver transplantation for glycogen storage disorders in children.
Liver transplantation (LT) may be indicated in glycogen storage disorders (GSD) when medical treatment fails to control the metabolic problems or when hepatic adenomas develop. We present our institutional experience with living donor LT (LDLT) for children with GSD. A total of 244 patients underwent primary LDLT at our institution from June 1994 to December 2005. A total of 12 (5%) children (8 female and 4 male) were afflicted with GSD and were not responsive to medical treatment. Nine patients had GSD type I and 3 had GSD type III. The median age at the time of transplantation was 7.27 yr (range, 2.4-15.7). All patients presented with metabolic abnormalities, including hypoglycemia, and lactic acidosis. In addition, 4 patients presented with growth retardation. A total of 11 patients received left lobe grafts and 1 received a right lobe graft. The mean graft-to-recipient weight ratio was 1.25 (range, 0.89-1.61). Two patients had hepatic vein stenoses that were treated by balloon dilatation; 1 patient had bile leak, which settled spontaneously. The overall surgical morbidity rate was 25%. Three patients had hepatic adenomas in the explanted liver. There was a single mortality at 2 months posttransplantation due to acute pancreatitis and sepsis. The mean follow up was 47.45 months. The metabolic abnormalities were corrected and renal function remained normal. In patients with growth retardation, catch-up growth was achieved posttransplantation. In conclusion, LDLT is a viable option to restore normal metabolic balance in patients with GSD when medical treatment fails. Long-term follow-up after LT for GSD shows excellent graft and patient survival.


[T2-dependent sequences in the study of hepatic focal lesions: comparison of the conventional spin echo sequence and the 0.5 T fast spin echo].
T2-weighted spin echo MR images are widely used in the detection and characterization of focal liver lesions. The main pitfall of this technique is its relatively long acquisition time. Fast spin echo sequences can provide the same contrast as conventional T2-weighted SE images in a shorter scanning time. Our study prospectively compared the effectiveness of T2-weighted conventional spin echo (CSE) versus fast spin echo (FSE) sequences in MRI of focal liver lesions. We examined 24 patients with this condition. All lesions were submitted to percutaneous biopsy and to the diagnostic gold standard technique, which was intraoperative US (13 patients), Lipiodol CT (10 patients) or 6-month follow-up (1 patient). MR images were analyzed from both a quantitative (signal-to-noise and contrast-to-noise ratios) and a qualitative viewpoints (overall image quality, lesion detection rate, number of lesions as compared with the gold standard, lesion conspicuity, internal features and the absence of artifacts). There was no statistically significant difference in contrast-to-noise ratio between the two sequences (p = 0.713). In the qualitative analysis, CSE sequences were superior to FSE for overall image quality in 50% of cases, for lesion conspicuity in 41.5% of cases and for internal features and the absence of artifacts in 46% of cases, FSE sequences had a higher detection rate in 17% of cases, even though both types of sequences underestimated the number of lesions in 29% of cases, as compared with the gold standard. To conclude, FSE sequences were inferior to CSE for image quality, lesion conspicuity, internal features and the absence of artifacts. FSE sequences were superior in the detection and characterization of fluid and nearly fluid lesions.


[Endocrine structures of the kidney in hypertension].
Comparative morphometric study of the juxtaglomerular complex and interstitial cells of the corticomedullary zone of the kidney was performed on the autopsy material in benign (26 cases) and malignant hypertension; for control, 15 cases were examined. The state of intrarenal arteries, arterioles and the degree of medulla interstitium sclerosis were taken into consideration. No activation of the renin-producing system was observed morphometrically in "benign" hypertension while it was pronounced in malignant hypertension. The hypothesis is put forward on the reserve renin secretion by mesangial cells in malignant hypertension. It is assumed that the prostaglandine-synthesizing function of the interstitial cells is not damaged in benign hypertension, but it is significantly lower in malignant course of the disease. Relationship is shown between the alterations of intrarenal arteries and arterioles and the degree of the medulla sclerosis in malignant hypertension.


Bayesian applications of belief networks and multilayer perceptrons for ovarian tumor classification with rejection.
Incorporating prior knowledge into black-box classifiers is still much of an open problem. We propose a hybrid Bayesian methodology that consists in encoding prior knowledge in the form of a (Bayesian) belief network and then using this knowledge to estimate an informative prior for a black-box model (e.g. a multilayer perceptron). Two technical approaches are proposed for the transformation of the belief network into an informative prior. The first one consists in generating samples according to the most probable parameterization of the Bayesian belief network and using them as virtual data together with the real data in the Bayesian learning of a multilayer perceptron. The second approach consists in transforming probability distributions over belief network parameters into distributions over multilayer perceptron parameters. The essential attribute of the hybrid methodology is that it combines prior knowledge and statistical data efficiently when prior knowledge is available and the sample is of small or medium size. Additionally, we describe how the Bayesian approach can provide uncertainty information about the predictions (e.g. for classification with rejection). We demonstrate these techniques on the medical task of predicting the malignancy of ovarian masses and summarize the practical advantages of the Bayesian approach. We compare the learning curves for the hybrid methodology with those of several belief networks and multilayer perceptrons. Furthermore, we report the performance of Bayesian belief networks when they are allowed to exclude hard cases based on various measures of prediction uncertainty.


Persistent candidemia in neonatal care units: risk factors and clinical significance.
The prevalence and clinical significance of persistent candidemia among neonates are poorly understood. This study aimed to describe the rate and the clinical relevance of persistent candidemia over a 4-year period in Kuwait. A retrospective chart review of infants admitted to the Neonatal Care Unit of the Maternity Hospital in Kuwait between January 2007 and December 2010, who had a positive blood culture for Candida species, was conducted. Persistent candidemia was defined as the isolation of the same Candida species more than 6 days after the initiation of antifungal therapy, or death due to candidemia within 6 days of antifungal treatment. Stepwise logistic regression was used to investigate factors associated with persistent candidemia. Of 89 neonates with a Candida infection, 54 (60.7%, 95% confidence interval 49.7-70.9%) had persistent candidemia. The case-fatality rate was 54% among those with persistent candidemia and 3% among those with non-persistent candidemia (p<0.001). Neonates with persistent candidemia were more likely to be female, have a central vascular catheter at diagnosis, and have a low platelet count. All isolated Candida species were susceptible to antifungal agents. Persistent candidemia is common among neonates with a Candida infection and is associated with an increased risk of mortality. Drug resistance is unlikely to explain the persistent candidemia; host-related factors seem to be more important and hence could be used to identify those at risk in order to institute appropriate preventive and treatment measures.


Concentrations, sources, and spatial distribution of individual polycyclic aromatic hydrocarbons (PAHs) in agricultural soils in the Eastern part of the EU: Poland as a case study.
Soils from agricultural areas receive unsatisfactory attention as regards the contamination with organic pollutants. To answer those needs the contents of the sixteen individual PAH compounds were determined (GC/MS technique) in agricultural soils in Poland. The samples (n=216) were collected from the upper layer of arable land in the year 2005. Half of the samples represented typical rural areas, while the rest derived from the territories potentially subjected to the urban/industrial pressure of various intensity. The mean (geometric) content of individual compounds varied from 1 microg kg(-1) for acenaphtylene to 55 microg kg(-1) for fluoranthene with the highest contributions (11.6%-12.9%) of phenanthrene, fluoranthene and pyrene. Higher molecular weight PAHs (4 rings) were strongly linked mutually and with the summation operator 16PAHs. They contributed substantially (73%) to the overall content of PAHs, which implies domination of anthropogenic sources. The calculated molecular indexes suggest that most of those PAHs derive from the combustion of coal, the main energy source in Poland. Simultaneously, the concentrations of lower molecular weight compounds seem to reflect the background, "natural" PAH compounds, which represent mainly atmospherically distributed emission. The division of the samples into groups describing geographical regions and landscape type enabled evaluation of the spatial trends in contamination of soils with PAH compounds. The most pronounced effect of spatial parameters corresponded to PAHs >4 rings, while lower molecular weight compounds showed more homogeneous concentration through the country.


CAD-associated reader error in CT colonography.
Computed tomographic colonographic interpretation with computer-aided detection (CAD) may be superior to unaided viewing, although polyp characteristics may influence accuracy. Reader error due to polyp characteristics was evaluated in a multiple-case, multiple-reader trial of computed tomographic colonography with CAD. Two experts retrospectively reviewed 52 positive cases (74 polyps) and categorized them as hard, moderate, or easy to detect. Each case was evaluated without and with CAD. Features that may influence a reader's ability to detect a polyp or to accept or reject a CAD mark were tabulated. The association between polyp characteristics and detection rates in the trial was assessed. The difference in detection rates (CAD vs unassisted) was calculated, and regression analysis was performed. Of 64 polyps found by CAD, experts categorized 20 as hard, 28 as moderate, and 16 as easy to detect. Reader characterization errors predominated (47.3%) over other errors. Factors associated with lower detection rates included small size, flat morphology, and resemblance to a thickened fold. CAD was superior for polyps resembling lipomas compared to those that did not resemble lipomas (average increase in detection rate with CAD, 12.8% vs 5.5%; P < .05). Polyp characteristic may impair computed tomographic colonographic interpretation augmented by CAD. Readers can avoid errors of measurement by evaluating diminutive polyp candidates with sample measurements. Caution should be taken when evaluating focally thick folds and when using visual impression to dismiss a polyp candidate as a lipoma when it is submerged in densely tagged fluid.


Abnormal subsurface hydrogen diffusion behaviors in heterogeneous hydrogenation reactions.
Hydrogen adsorption and diffusion behaviors on noble metal model catalyst surfaces and into the subsurfaces are of paramount significance in the exploration of novel heterogenous catalytic hydrogenation reactions. We present an in-depth study of hydrogen adsorption on and diffusion into the subsurfaces of three typical 5d noble metals from three-dimensional electronically adiabatic potential energy surfaces (PESs) by interpolating plenty of ab initio density functional theory (DFT) configuration-energy points. The surfaces and subsurfaces regions of the relaxed Ir(100) and (111), Pt(100) and (111), and Au(100) and (111) surfaces, are, respectively, taken into account. For hydrogen adsorption on the (100) surfaces, the lowest adsorption energy site is the Bridge site, instead of the traditional Hollow site. Hydrogen prefers to follow the indirect pathway with a lower diffusion barrier, in the competition with the direct pathway with much higher diffusion barrier. For hydrogen diffusion on the (111) surfaces, hydrogen follows the pathway from Top site to fcc site on the surface and prefers an up-down direct pathway into the subsurface. Importantly, the nudged elastic band (NEB) based on the PESs can reproduce those results calculated from the NEB(DFT) very well. The developed highly-accurate and efficient approach based on the PESs helps us to further investigate the more complex reactant diffusion dynamics at surfaces.


Expression of heme oxygenase-1 in rat ontogeny.
Heme oxygenase (HO), the heme-degrading enzyme, plays an important role in heme catabolism. Among three isozymes, HO-1 is an inducible form expressed mainly in macrophages. In rat ontogeny, HO-1 immunoreactivity was detected in mononuclear cells in the yolk sac at 10 days of gestation. HO-1-expressing cells were then detected in the fetal liver and their numbers increased during the gestational period. The numbers of HO-1-positive cells and HO-1 mRNA levels in the liver peaked at 18 days of gestation. Most of the macrophages expressed both HO-1 and a macrophage scavenger receptor. Macrophages in the fetal liver showed marked hemophagocytosis. Macrophages in the lung, spleen, bone marrow, and other tissues also expressed HO-1. HO-1 immunoreactivity was also observed in syncytial cells of the chorionic villi, the endodermal layer of the yolk sac, and renal tubules of the fetus. Intestinal mucosal epithelial cells expressed HO-1 after birth. These findings imply that HO-1 is crucial for macrophages in heme catabolism from an early stage of ontogeny. HO-1 expression in non-macrophagic cells may be required for other purposes such as protection from oxidative stress and various stimuli.


Reciprocal EMG control of elbow extension by FES.
Elbow extension is critical in performing activities of daily living. Individuals with a C5-C6 spinal cord injury have paralyzed elbow extensors, yet retain weak to strong voluntary control of elbow flexion. Previous studies have shown that functional electrical stimulation (FES) of the triceps provides sufficient elbow extension strength and control to greatly improve function. With triceps stimulation applied at a constant level, elbow angle is controlled naturally by voluntary flexion opposing the stimulated extension-referred to as voluntary antagonist control. We have investigated an alternative reciprocal control scheme employing biceps electromyogram (EMG) to modulate triceps stimulation. With reciprocal control, increasing biceps EMG proportionally reduces triceps stimulation. A personal computer (PC)-based lab system was designed to test the feasibility of reciprocal control. Reciprocal control increased the range of elbow moments, was stable during maintained elbow angle or isometric moment, and used less stimulation. Reciprocal control of triceps stimulation using biceps EMG is an effective method for restoring elbow extension to C5-C6 spinal cord injury patients, and could be extended to other situations where a voluntarily controlled muscle can be opposed by stimulating an antagonist.


[The action of fenasal on the cestode Diphyllobothrium latum].
The present study used an experimental invasion model to examine the invasion and behaviour of D.latum cestoda and its tissue structural disorders after in vivo phenasal administration. Phenasal caused an increased development of the granular endoplasmic reticulum in the cells of the tegument and yolk glands, their local spread, and a formation of vacuoles in the tegumental cells and an appearance of myelins which were rather great in size. There were no changes in tegumental microtrichia.


Microextraction techniques in the analysis of food flavor compounds: A review.
Food flavor compounds due to the complexity of food as a matrix, and usually their very low concentrations in a product, as well as their low odor thresholds, create a challenge in their extraction, separation and quantitation. Food flavor volatiles represent compounds of different polarity, volatility and chemical character, which determine method of extraction for their isolation from food. Microextraction techniques, mainly SPME and SBSE have been used for food flavor compounds analysis for two decades. Microextraction methods other than SPME and SBSE are seldom used despite their analytical potential. The review discusses the nature of food flavor compounds, and different approaches to food flavor analysis. It summarizes the use of microextraction methods in food flavor compounds analysis based on papers published in the last 5years, and discusses the potential of microextraction methods in this field.


Perceptions of slow codes by nurses working on internal medicine wards.
Cardio-pulmonary resuscitation is the default procedure during cardio-pulmonary arrest. If a patient does not want cardio-pulmonary resuscitation, then a do not attempt resuscitation order must be documented. Often, this order is not given; even if thought to be appropriate. This situation can lead to a slow code, defined as an ineffective resuscitation, where all resuscitation procedures are not performed or done slowly. To describe the perceptions of nurses working on internal medicine wards of slow codes, including the factors associated with its implementation. This was a cross-sectional, descriptive study. Participants completed a personal characteristics questionnaire and the Perceptions and Factors of Slow Codes questionnaire designed for this study. Participants and research context: The sample was a convenience sample of nurses working on internal medicine wards in two Israeli hospitals. Ethical considerations: The study received ethical approval from both institutions, where data were collected and stored according to institutional policy. Most reported that resuscitations were conducted according to protocol (n = 90, 76.2%). Some took their time calling the code (n = 22, 18.3%), or waited by the bedside and did not perform cardio-pulmonary resuscitation (n = 45, 37.5%). Factors most associated with slow codes were poor patient prognosis (mean = 3.52/5, standard deviation = 1.27) and a low chance of patient survival (mean = 3.37/5, standard deviation = 1.21). Two-thirds (n = 76, 66.8%) reported that slow codes were done on their unit and the majority (n = 80, 69%) perceived slow codes as ethical. This study confirms that slow codes are part of medical care on internal medicine wards, where most nurses perceive them as an ethical alternative. These perceptions are in contrast to most legal and ethical opinions expressed in the literature. Nurses should be educated about the legal and ethical implications of slow codes, and qualitative and quantitative studies should be conducted that further investigate its implementation.


Effects of time and an educational presentation on student attitudes toward wife-beating.
To determine if a public outreach educational presentation on domestic violence would produce change in attitudes toward wife-beating and if the attitude change would persist over time, 36 female and male university students were quasi-randomly assigned to groups. Half of the subjects received a 20-minute presentation on domestic violence (Group P), and half did not receive a presentation (Group NP). Groups were then tested on their responses on the Inventory of Beliefs about Wife Beating (IBWB). Three weeks later, groups were retested on their responses to the scale. MANOVA revealed that attitudes of Group P were significantly more negative toward wife-beating than attitudes of Group NP on three of the five IBWB subscales and on three of the 31 IBWB items subsumed under the subscales. Data supported the prediction that public outreach efforts would produce more negative attitudes toward wife-beating and that attitude change would not deteriorate over time.


Antimicrobial susceptibility of viridans group streptococci.
A total of 68 viridans group streptococci, including 31 Streptococcus sanguis, 12 S. mitis, 3 S. salivarius, and 8 S. milleri from blood, and an additional 14 S. milleri from abscesses and normally sterile sites, were tested against penicillin, amoxicillin, cefazolin, ceftriaxone, meropenem, clindamycin, quinupristin/dalfopristin, rifampin, levofloxacin, ofloxacin, vancomycin, and gentamicin with the microdilution method. The susceptibility rates for S. sanguis were: penicillin, 74%; amoxicillin, 84%; ceftriaxone, 94%; clindamycin, 87%, and vancomycin, 100%. The susceptibility rates for S. mitis were: penicillin, 42%; amoxicillin, 67%; ceftriaxone, 58%; clindamycin, 100%; and vancomycin, 100%. The susceptibility rates for S. milleri were: penicillin, 100%, amoxicillin. 100%; ceftriaxone, 100%, clindamycin, 100%; and vancomycin, 100%. Two of the three isolates of S. salivarius were susceptible to penicillin, amoxicillin, and ceftriaxone; all were susceptible to clindamycin and vancomycin. Levofloxacin, quinupristin/dalfopristin, and rifampin were highly active against all isolates.


Lack of effective translational regulation of PLD expression and exosome biogenesis in triple-negative breast cancer cells.
Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer that is difficult to treat since cells lack the three receptors (ES, PR, or HER) that the most effective treatments target. We have used a well-established TNBC cell line (MDA-MB-231) from which we found evidence in support for a phospholipase D (PLD)-mediated tumor growth and metastasis: high levels of expression of PLD, as well as the absence of inhibitory miRs (such as miR-203) and 3'-mRNA PARN deadenylase activity in these cells. Such findings are not present in a luminal B cell line, MCF-7, and we propose a new miR•PARN•PLD node that is not uniform across breast cancer molecular subtypes and as such TNBC could be pharmacologically targeted differentially. We review the participation of PLD and phosphatidic acid (PA), its enzymatic product, as new "players" in breast cancer biology, with the aspects of regulation of the tumor microenvironment, macrophage polarization, regulation of PLD transcripts by specific miRs and deadenylases, and PLD-regulated exosome biogenesis. A new signaling miR•PARN•PLD node could serve as new biomarkers for TNBC abnormal signaling and metastatic disease staging, potentially before metastases are able to be visualized using conventional imaging.


[Identification of differentially expressed genes of acute hypoxia-treated HepG2 cells and hypoxia-acclimatized HepG2 cells].
To provide necessary information for further understanding of molecular mechanism of hypoxia acclimatization, the differentially expressed genes of HepG2 cells exposed to normoxia, acute hypoxia-treated cells which were exposed to 1% oxygen for 48 h, and hypoxia-acclimatized HepG2 cells which were cultured for 6 circles of alternate low oxygen (1% oxygen for 24 h) and normal oxygen (21% oxygen for 24 h), were identified respectively by combining the suppression subtractive hybridization (SSH) and cDNA microarray. Thirty-seven genes were expressed differentially in cells exposed to 1% oxygen for 48 h compared with those in cells exposed to normoxia. The expression of all these 37 genes was down-regulated, including the genes participating in cell cycle, cell response to stimulus, and cell signal transduction, and cell cytoskeleton formation, the genes associated with transcription and cell metabolism, 4 expressed sequence tags (ESTs), and 12 genes of which the functions are not known. There is a novel gene sequence, which has not been found in existing databases. There were only 6 genes differentially expressed in the hypoxia-acclimatized cells compared with cells exposed to normoxia, including two mitochondrion genes, metalloprotease-1 gene, ferritin gene, thymosin beta-4 and TPT1 genes. The expressions of mitochondrion ND4, ferritin, thymosin beta-4 and TPT1 were up-regulated, while the expressions of mitochondrion ND1 gene and metalloproease-1 gene were down-regulated. Cell tolerance to hypoxia increased after the cells were hypoxia-acclimatized. The different gene expression patterns of the acute hypoxia-treated cells and the hypoxia-acclimatized cells may be related to the increased tolerance of the cells to hypoxia.


Expression of synthetic genes coding for completely new, nutritionally rich, artificial proteins.
Synthetic genes (A, AB and AHB) constructed and cloned into pKK233-2 vector were recloned from the parent plasmid into the new procaryotic expression vectors pGFY221N and pBI052. Gene AF-B (coding for all amino acids besides phenylalanine) was obtained by 'cassette mutagenesis' from gene AB. The plasmid pGFY221N was constructed from pGFY218L by replacing the PstI by an NcoI site; plasmid pBI052 was derived from pGFY221N through replacing the 221-bp EcoRI/NcoI fragment with a synthetic DNA segment of 52 bp representing the Escherichia coli atpE gene translational initiation region. The genes A, AB, AHB and AF-B in the vector pGFY221N were expressed with a six-amino-acid-long leader sequence; in pBI052 the genes were expressed directly. In vitro expression experiments were successfully with all the genes except with the AHB gene integrated into pGFY221N. In the E. coli minicell system expression was demonstrated with the A gene in pGFY221N and the AF-B and AHB genes in pBI052. Complete translation of the expressed genes AB, AF-B and AHB in either the in vitro or in vivo systems could be shown by using 35S-labelled N-terminal methionine and C-terminal cysteine. Both amino acids occur only once in the peptide sequences.


Structural and mutational analysis of substrate complexation by anthranilate phosphoribosyltransferase from Sulfolobus solfataricus.
The metabolic synthesis and degradation of essential nucleotide compounds are primarily carried out by phosphoribosyltransferases (PRT) and nucleoside phosphorylases (NP), respectively. Despite the resemblance of their reactions, five classes of PRTs and NPs exist, where anthranilate PRT (AnPRT) constitutes the only evolutionary link between synthesis and degradation processes. We have characterized the active site of dimeric AnPRT from Sulfolobus solfataricus by elucidating crystal structures of the wild-type enzyme complexed to its two natural substrates anthranilate and 5-phosphoribosyl-1-pyrophosphate/Mg(2+). These bind into two different domains within each protomer and are brought together during catalysis by rotational domain motions as shown by small angle x-ray scattering data. Steady-state kinetics of mutated AnPRT variants address the role of active site residues in binding and catalysis. Results allow the comparative analysis of PRT and pyrimidine NP families and expose related structural motifs involved in nucleotide/nucleoside recognition by these enzyme families.


Successful clot lysis using low dose of streptokinase in 22 neonates with aortic thromboses.
To determine whether intravenous infusion of low dose of streptokinase was effective in lysing umbilical arterial catheter (UAC)-associated aortic thrombi. A prospective cohort study of 31 consecutive newborn infants with UAC-associated aortic thrombi which were detected by abdominal ultrasonography after removal of UAC. Twenty-two infants were treated with intravenous infusion of low dose (1000 U/h) streptokinase, while nine others were not treated due to various contra-indications. Thrombolysis occurred after a mean interval of 2.2 days (standard deviation (SD) = 1.8) in the treated infants. In the untreated infants, spontaneous thrombolysis occurred significantly later, after a mean interval of 16.9 days (SD = 14.7) (95% confidence intervals of difference between mean intervals - 26.0, - 3.4; P = 0.02). Only one treated infant developed mild bleeding directly attributed to streptokinase therapy. Low dose streptokinase infusion was effective and safe in thrombolysing UAC-associated aortic thrombi.


The moderating role of interviewer's regulatory focus in the effectiveness of impression management tactics: Regulatory fit as a source of subjective value.
Based on the recently explored regulatory fit effect in social contexts, the present research is the first to investigate the interaction between interviewer's regulatory focus and interviewee's impression management (IM) tactics. We hypothesized that assertive and defensive IM tactics would fit with interviewer's promotion and prevention focus, respectively, and that interviewer's experience of this regulatory fit would lead to enhanced interview evaluation. We conducted four studies in which the participants were asked to rate an interviewee after reading a list of the interviewee's IM-related behaviours or watching a videotaped interview. Additionally, the participant's regulatory focus was operationalized as both an induced situational state and a measured chronic trait. The results supported our hypotheses by showing a significant interaction of the interviewer's regulatory focus and the IM tactics on interview evaluation: the promotion-assertive and the prevention-defensive ratings were more positive than the prevention-assertive and the promotion-defensive ratings. Moreover, mediation analysis revealed that the interaction exerts its effects on interview evaluation by providing interviewers with an intrapersonal regulatory fit experience.


Pain complaints as risk factor for mental distress: a three-year follow-up study.
The main aim of this study was to investigate the effect of pain complaints at baseline and mental distress at follow-up. We included adolescents from two longitudinal studies of 18/19 years of age with two corresponding cross-sectional baseline studies at age 15/16. A total of 5750 were invited for the baseline study, and we have 3-year follow-up data for 3316 (57.8% follow-up rate). All information is based on self-report. The questions of pain are asking for severe pain the past year. Mental distress is measured by Hopkins Symptoms Check List (HSCL-10). Number of pain sites increased among girls and decreased among boys from baseline to follow-up. There was a cross-sectional dose-response association between number of pain sites and mental distress at both time points. Also, in the follow-up study, there was dose-response relationship between numbers of pain sites at baseline and mental distress at follow-up even after controlling for possible confounding factors. We found no effect of mental distress at baseline on pain reports at follow-up. There is a strong association between pain at 15/16 years and mental distress at 18/19 years of age. Clinicians should therefore take pain complaints among adolescents seriously and be aware of comorbid and later development of mental health problems.


Comparison of Isolab Quik-Sep and Quik-Sep System II for separation of immunoglobulins G and M.
Use of Quik-Sep System II (QSII) ion-exchange chromatography increased the detection of CMV-directed immunoglobulin M (IgM) compared to the prototype Quik-Sep System (QSI) (27 of 87 [31%] IgM-positive versus 20 of 87 [24%]). However, an increase in the number of false-positive rheumatoid factor-containing sera was found (15/52 [29%] versus 8/52 [15%]). Total immunoglobulin G (IgG) and IgM concentrations were higher in the QSII eluates compared to those from QSI, although total IgM recovery did not exceed 36% in either system.


Long-term registration of cutaneous microcirculation during general anesthesia.
The temporal dynamics of the systemic arterial pressure can be monitored noninvasively from the skin of the earlobe or forehead by photoplethysmography under the provision that the active control of the microcirculatory perfusion is eliminated. Using this approach, we have been able to detect a highly stable blood pressure rhythm in the range of 0.15 Hz during psychophysical relaxation or sleep. The aim of the present study was to investigate the occurrence and behavior of blood pressure rhythms below 0.2 Hz during general anesthesia. In 30 patients (ASA groups I-II) undergoing basic surgical procedures, photoplethysmographic recordings from the earlobe were made during the whole time of anesthesia. The recorded signals were divided into segments of 200 s of duration, the temporal structure of which was analyzed by fast Fourier transform. Different characteristic patterns of rhythmical behavior were detected: (1) absence of activity below 0.2 Hz ('low-frequency range'); (2) slow sinusoidal rhythmicity below 0.05 Hz; (3) 'chaotic' behavior, i.e. multiple incoherent fluctuations without stationary periods or amplitudes; (4) short-term rhythmical activity at about 0.15 Hz, and (5) long-term rhythmical activity at about 0.15 Hz. In patients sufficiently sedated to eliminate low-frequency activity, rhythmicity could sometimes be triggered by certain surgical stimuli, the response to which was suppressed by injection of opioids. The data presented strongly suggest that rhythmical perfusion patterns of the cutaneous microcirculation could serve as an indicator for the depth of anesthesia.


CYP2B2 gene expression and phenobarbital induction in kidneys using an in vitro transcription system.
The induction of cytochrome P450 2B1 and 2B2 in rat liver in response to phenobarbital is mediated at the transcriptional level. While in extrahepatic tissue such as the kidney, the transcription level of the CYP 2B2 gene is much lower than in the liver. The transcriptional activity of the CYP2B2 promoter constructs in nuclear proteins from kidney revealed that the in vitro system maintained the tissue-specificity of CYP2B2 transcriptional induction but not that of the basal transcription of the gene.


Prenatal exposure to ethanol induces leptomeningeal heterotopia in the cerebral cortex of the rat fetus.
Pregnant rats were fed an ethanol-containing liquid diet between gestational day (GD) 10 and GD 21. Leptomeningeal heterotopias were observed in the cerebral cortex of ethanol-exposed fetuses. They appeared on the brain surface of the lateral cortical region near the rhinal fissure, and were found more numerously in the rostral than the caudal region. These abnormalities contained certain neuronal perikarya, microtubule-associated protein (MAP) 1b-positive neuronal processes, and Rat-401-positive radial glial fibers. Immunostaining for Rat-401 revealed that the heterotopias protruded through breaches in the glia limitans. In adult rats exposed to ethanol prenatally, the heterotopias persisted in the lateral cortical region. We conclude that prenatal exposure to ethanol might induce defects in the glia limitans, resulting in the genesis of leptomeningeal heterotopias. These abnormalities may be related to mental retardation or the cognitive deficits associated with human fetal alcohol syndrome (FAS).


Increased hyaluronan flux from skin following burn injury.
Hyaluronan (formerly hyaluronic acid) is an important constituent of the interstitial matrix in skin. Following major burn injury in animal models, plasma hyaluronan can increase to levels 10-fold greater than normal. The present experiments were designed to determine whether this is a result of the increased lymph flow (QL) accompanying the injury or of an increased release of hyaluronan from the burned skin and subcutaneous tissue. The lateral saphenous vein and a prenodal lymphatic were cannulated in the hindpaw of five anesthetized canines. Hindpaw venous pressure was elevated until the total protein concentration in lymph declined to steady-state levels, and QL and hyaluronan flux (QL X [hyaluronan]) was measured. A minor burn was inflicted by immersion of the paw into 100 degrees C water for five sec, and measurements were repeated at regular intervals for a minimum of 4 hr. Burn injury resulted in significant and persistent increases in QL (154 +/- 61 microliters/min versus 562 +/- 105 microliters/min 4 hr postburn) and lymph total protein concentration (1.34 +/- 0.04 g/dl versus 4.08 +/- 0.18 g/dl 4 hr postburn), while lymph hyaluronan concentration fell (3.01 +/- 0.20 micrograms/ml versus 2.1 +/- 0.16 micrograms/ml 4 hr postburn). The resultant increase in hyaluronan flux (0.42 +/- 0.13 microgram/min versus 1.17 +/- 0.22 microgram/min 4 hr postburn) appears to be a function of lymph flow rather than burn-induced release of skin hyaluronan. Hence, the increased plasma concentration of hyaluronan following major burns is likely a consequence of increased lymph flow from the site of injury.


[The causes of prolonged fever in patients with infiltrative lung processes].
The authors analyze the causes of lingering fever in patients with pulmonary infiltrations. The given phenomenon is most often caused by pneumonias provoked by unusual causative agents (Legionella, Rickettsia, Mycoplasma). In such cases, administration of erythromycin is effective. In rare cases, lingering fever is induced by blocked pulmonary suppurations, pleural exudates in pneumonia patients, and infiltrative tuberculosis. Besides, there were cases, in which fever was of drug etiology.


Metabolism of gemcitabine in rat and dog.
1. The metabolism of 14C-gemcitabine in the male rat has been studied after intravenous administration of a single dose (10 mg/kg) or five doses (1 mg/kg/day) of 14C-gemcitabine. The metabolism in male dog has been studied after only single dosing. The effects of gemcitabine on hepatic drug-metabolizing enzymes in rat has also been studied. 2. The concentration of gemcitabine in the plasma was 11.84 micrograms/ml at 5 min, and then rapidly decreased after a single administration to rat. A deaminated uracil analogue of gemcitabine progressively increased with time. Gemcitabine and the uracil metabolite accounted for 80.0 and 11.8% of the radioactive dose in the 0-24-h urine samples respectively. Gemcitabine was the major component identified in lung, liver and kidney at 5 min after administration. 3. After repeated administration to rat, metabolites in the plasma and tissues were not remarkably different from those found after a single administration. 4. After a single administration to dog, the plasma concentration of gemcitabine was 12.39 micrograms/ml at 5 min. Gemcitabine and the uracil metabolite accounted for 8.3 and 71.8% of the dose in the 0-24-h urine samples respectively. 5. No differences were observed in enzymatic activities per whole liver between the gemcitabine-treated and control rat.


Bifid median nerve with three thenar branches--case report.
A case of a high division of the median nerve associated with a persistent median artery and three transligamentous thenar motor branches is presented. To our knowledge, three transligamentous motor branches have not been previously reported in the literature.


Cronkhite–Canada syndrome six decades on: the many faces of an enigmatic disease.
Cronkhite–Canada syndrome is a rare gastro-enterocolopathy of uncertain aetiology first described almost 60 years ago. It is characterised by diffuse gastrointestinal polyposis sparing only the oesophagus, ectodermal abnormalities and an unpredictable but often fatal clinical course. The disease may demonstrate extremely diverse clinical and endoscopic features, which often leads to a delay in diagnosis. A high index of suspicion and recognition of the characteristic histological findings frequently facilitate a correct diagnosis, but the distribution of the gastrointestinal pathology and its microscopic features may be atypical. The pathologist thus requires a thorough knowledge of both the typical and many atypical faces of this disease, for which various documented therapies often still prove ineffective. Close correlation with clinical findings, including any pertinent ectodermal abnormalities, and careful examination of biopsies derived from polypoid and endoscopically spared mucosa will ensure a timely and correct diagnosis in patients with this enigmatic syndrome.


Vertical rectus muscle transposition and botulinum toxin for complete sixth nerve palsy.
Effective surgical treatment of complete unrecovered sixth nerve palsy must include the transfer of abducting power to the temporal aspect of the globe with release of medial rectus contracture nasally. We describe our experience in the treatment of five such patients who underwent full vertical rectus transposition combined with botulinum toxin chemodenervation of the ipsilateral medial rectus muscle. The five patients all had primarily unilateral complete unrecovered sixth nerve palsy. They all underwent a complete preoperative and postoperative eye examination and an orthoptic assessment. Excursion into abduction was graded from -8 (globe immobilized in extreme adduction) to -4 (abduction as far as primary position) to 0 (full abduction). Abduction saccades and a forced muscle generation test confirmed the presence of complete unrecovered sixth nerve palsy, and forced duction testing measured the degree of medial rectus contracture. All patients received ipsilateral medial rectus injection of botulinum toxin in the preoperative (8 to 2 months before surgery) and perioperative periods, and underwent complete superior rectus-inferior rectus transposition temporally. The average length of follow-up was 21 (range 6 to 48) months. The average preoperative distance alignment was 52 (range 25 to 80) prism dioptres (PD). Vertical rectus transposition combined with botulinum toxin injection resulted in an average distance alignment change of 66 PD (range 50 PD to 82 PD) of exoshift. The average final deviation was 1 PD of esotropia (range 4 PD of esotropia to 6 PD of exotropia). Average abduction improved from -6 (range -3 to -8) preoperatively to -1.7 (range -1 to -2) postoperatively. Saccades averaged -4 preoperatively and improved to -2 postoperatively. Normal vertical eye movements were preserved in all patients. A total field of single binocular vision was created in all patients, which averaged 55 degrees (range 30 degrees to 75 degrees) in the horizontal meridian. The field of single binocular vision from primary position into abduction averaged 23 degrees (range 18 degrees to 28 degrees). Temporal transposition of the vertical rectus muscles combined with perioperative botulinum toxin injection of the ipsilateral medial rectus muscle is a reliable and effective way of restoring functional binocular vision in patients with complete unrecovered sixth nerve palsy.


Performance of spinal cord injury individuals while standing with the Mohammad Taghi Karimi reciprocal gait orthosis (MTK-RGO).
Most patients with spinal cord injury use a wheelchair to transfer from place to place, however they need to stand and walk with orthosis to improve their health status. Although many orthoses have been designed for paraplegic patients, they have experienced various problems while in use. A new type of reciprocal gait orthosis was designed in the Bioengineering Unit of Strathclyde University to solve the problems of the available orthoses. Since there was no research undertaken regarding testing of the new orthosis on paraplegic subjects, this study was aimed to evaluate the new orthosis during standing of paraplegic subjects. Five paraplegic patients with lesion level between T12 and L1 and aged matched normal subjects were recruited into this study. The stability of subjects was evaluated during quiet standing and while undertaking hand tasks during standing with the new orthosis and the knee ankle foot orthosis (KAFO). The difference between the performances of paraplegic subjects while standing with both orthoses, and between the function of normal and paraplegic subjects were compared using the paired t test and independent sample t test, respectively. The stability of paraplegic subjects in standing with the new orthosis was better than that of the KAFO orthosis (p < 0.05). Moreover, the force applied on the crutch differed between the orthoses. The functional performance of paraplegic subjects was better with the new orthosis compared with normal subjects. The performance of paraplegic subjects while standing with the new orthosis was better than the KAFO. Therefore, the new orthosis may be useful to improve standing and walking in patients with paraplegia.


Agonist selective modulation of tyrosine hydroxylase expression by cannabinoid ligands in a murine neuroblastoma cell line.
Functional interactions between catecholamines and cannabinoid transmission systems could explain the influence of Delta(9)-tetrahydrocannabinol on several central activities. Hence, the presence of cannabinoid CB(1) receptors in tyrosine hydroxylase (TH) containing cells has been suggested, providing clue for a direct control of catecholamines synthesis. In the present study, we evidenced the constitutive expression of functional cannabinoid CB(1) receptors in N1E-115 neuroblastoma and reported on the use of this model to examine the influence of diverse cannabinoid ligands on TH expression. Exposure of the cells to the high-affinity agonist HU 210 (5 h) resulted in a significant decrease in TH content (pEC(50): 6.40). In contrast, no change was observed after a similar treatment with the structurally unrelated agonist CP 55,940. Besides, the use of a luciferase reporter assay revealed that these two agonists showed opposite influences on TH gene promoter activity. Thus, in cells expressing pTH-luc constructs, inhibition and induction of luciferase activity were respectively observed with HU 210 (pEC(50): 8.95) and CP 55,940 (pEC(50): 9.09). Pharmacological characterisation revealed that these reciprocal responses were both related to the specific activation of cannabinoid CB(1) receptor, suggesting an agonist-dependent modulation of distinct signalling pathways. While these data points out the possible pharmacological manipulation of TH expression by cannabinoid ligands, such approach should take into account the existence of agonist selective trafficking of cannabinoid CB(1) receptor signalling.


Sodium and potassium measurements: direct potentiometry and flame photometry.
Data from the 1979 and part of the 1980 College of American Pathologists (CAP) Chemistry Survey Program have been reviewed in order to compare the direct potentiometric measurement of sodium and potassium with flame-emission photometric measurements. As may have been predicted from earlier intralaboratory evaluations, the results of the CAP interlaboratory comparison confirm that flame-emission photometry of the two electrolytes yields lower mean concentrations. Reasons for this difference lie in the physicochemical aspects of the sample and methods. In Survey specimens, the magnitude of difference was rarely one of clinical significance, but the documented increase in the use of direct electrode measurements in laboratories warrants informing clinicians of potentially significant differences in selected patients.


Haemopoietic tissue in Perinereis cultrifera Grübe: cytological identification and observations after treatment with lead.
A perivascular tissue made up of two different types of cells (one with a slightly basophilic cytoplasm and many Fe+++ ions, the other with haemopigments) has been demonstrated in Perinereis cultrifera Grübe by means of histochemical and cytochemical techniques. These two types of cells probably cooperate in the production of the respiratory pigment. Treatment with lead nitrate (0.02 ppm) for up to a maximum of five days caused a progressive cytological alteration with inhibition of haemoglobin synthesis.


Glycoforms of alpha 1-acid glycoprotein in sera of human immunodeficiency virus-infected persons.
In acute infections thus far studied, there is a relative increase in plasma protein glycoforms rich in biantennary complex type N-glycans (type I), while in some diseases with chronic inflammatory changes, there is increase in glycoforms with more branched N-glycans (type II). In sera of 109 human immunodeficiency virus (HIV)-infected persons, 38 rheumatoid arthritis patients, and 44 healthy subjects, the composition of alpha 1-acid glycoprotein (AGP) glycoforms was studied using crossed immunoaffinity electrophoresis with concanavalin A as a ligand. In patients in CDC classifications I, II, and III, distribution of AGP glycoforms was analogous to that in normal subjects. Type I alterations were observed in patients in group IV who had no signs of arthritis. Type II changes, analogous to those found in rheumatoid arthritis, were seen in group IV patients who developed arthritis. Most significant type I changes were associated with Pneumocystis carinii pneumonia (specificity, 100%; sensitivity, 96%).


Migration as a determinant of marriage pattern: preliminary report on consanguinity among Afghans.
Two sample populations, one refugee and one resident, were studied. The frequencies of consanguineous marriages came out to be 49.8%and 55.4%, respectively, for the refugees and the residents. Caste endogamy was dominant both in the residents and the refugees. The mean coefficient of inbreeding was calculated to be 0.0303 for the refugee population and 0.0332 for the resident population samples. First cousin marriage was the dominant type of marriage in both samples; father's brother's daughter (FBD) marriage was more frequent among the refugees while mother's brother's daughter(MBD) marriage was more frequent among the residents. Education has no decreasing effect on the incidence of consanguineous marriages. A significant difference in the pattern of marriages in the refugees is observed after the Saur Revolution of 1979.


A study of atherothrombotic biomarkers in welders.
Studies have shown that welders have increased cardiovascular mortality. This may be due to airborne particulate matter (PM) exposure. Elevated levels of PM in polluted urban air have been associated with increased cardiovascular mortality and morbidity. This study seeks to explore potential mechanisms for the increased cardiovascular mortality in welders. Seventy welders were compared to 74 referents. Exposure to PM was assessed by personal full-shift sampling of work room air the last 2 days before collection of blood samples. Selected biomarkers of pro-coagulant activity, endothelial/platelet activation and systemic inflammation were determined in the samples. The welders had been occupationally exposed to PM for 15 years on average. The geometric mean current exposure to PM was 8.1 mg/m3. They had statistically significantly higher concentrations of TNF-α, P-selectin, CD40L, prothrombin fragment 1 + 2 and D-dimer than the referents. Increasing concentrations of D-dimer and CD40L were observed by increasing current exposure to PM. The study shows that welders highly exposed to welding PM were in a pro-thrombotic state with increased thrombin generation and consequently higher D-dimer concentrations. The welders had also increased endothelial/platelet activation as compared to the referents. These alterations are compatible with increased cardiovascular mortality as previously reported among welders.


Very high frequency ultrasound biometry of the anterior and posterior chamber diameter.
To measure the largest diameter of the anterior chamber (AC) and posterior chamber (PC) dimension and its orientation and determine the relationship with the principal keratometric meridians. Twenty-eight eyes of 14 subjects were scanned with high frequency (50 MHz) ultrasound in sequential meridional scan planes at 30 degrees increments. Observer identified angle and ciliary sulcus recess boundaries in each patient scan set were fit with an elliptical model to obtain the ellipse semi-major axis corresponding to the largest diameter and its meridional orientation. Anterior and posterior chamber diameters from raw data and model fit were compared using linear statistics. Circular statistics were used to compare the orientation of the largest diameter for raw ultrasound measurements, model estimations of largest diameter, and autorefractor determined keratometric axes. The mean model diameters were anterior chamber OD 12.07 mm (0.32 SD); anterior chamber OS 12.06 mm (0.36 SD); posterior chamber OD 12.35 mm (0.42 SD); posterior chamber OS 12.33 mm (0.43 SD). The general trend for orientation of the meridian of largest diameter was in the horizontal meridian. In over 35% of eyes the difference between AC or PC meridian and the flat keratometric axis was greater than 20 degrees. Accurate and reproducible anterior segment biometry depends on visualization of structures and minimization of eye and head movement error. The range and standard deviation of the diameter and orientation measures suggests anatomic variation is sufficient to require biometry for proper sizing and placement of intraocular devices that use angle or sulcus fixation.


[Analysis of influencing factors on hepatic encephalopathy in patients with non-cirrhotic portal hypertension].
Objective: To investigate the incidence of hepatic encephalopathy (HE) in patients with non-cirrhotic portal hypertension (NCPH) and to explore its risk factors. Methods: The incidence rate of HE in 150 cases with NCPH was evaluated in two hospitals, and 188 cases of compensated cirrhosis patients were taken as control. Logistic regression was used to screen for independent risk factors for HE in patients with NCPH. Results: The incidence of overt hepatic encephalopathy (OHE) in patients with NCPH was not statistically significantly different from that in patients with cirrhosis (4.7% vs. 6.9%, P = 0.682). The incidence of mild hepatic encephalopathy (MHE) was significantly lower than that of cirrhosis patients (32.7% vs. 46.3%, P < 0.05). The presence of upper gastrointestinal bleeding, infection and portosystemic venous shunt were the main independent factors for HE in NCPH patients (OR > 1, P < 0.05). Conclusion: HE is one of the important complications of NCP, and may be influenced by factors such as upper gastrointestinal bleeding, infection and portosystemic venous shunt.


How host defense is encoded in the mammalian genome.
This issue of Mammalian Genome explores the genetic approach to infectious disease susceptibility as it has been applied in mammals. Although no single issue of any journal could give comprehensive treatment to a field so extensive and rapidly growing as this one, these texts describe key discoveries that provided new understanding of immune responses. Classical genetic studies opened and continue to pave the way to deep understanding of many issues in immunology.


Prognosis of replanted primary incisors after injuries.
Between 1979 and 1997, 58 avulsed primary teeth were treated at the Pedodontic Clinic of Niigata University Dental Hospital. Among these, we replanted six avulsed incisors of four patients and treated two teeth of two patients replanted at other dental clinics. Referring to the clinical records, oral photographs and radiographs, we examined the injury age, cause of injury, condition of tooth storage, length of time until replantation, and also the prognosis after replantation. Three teeth of two cases remained until eruption of their permanent successors, and one tooth of one case remained under observation without extraction. Although the other four teeth of three cases resulted in extraction, no secondary infection was detected due to replantation. The following reasons were suspected for the poor prognosis of the four teeth. One avulsed tooth was not fixed immediately after replantation. One replanted tooth might not have been compatible with the alveolar socket. In the other two teeth, the periodontal vital tissues might have been removed before replantation. It is generally suggested that replantation of primary teeth is not a good option. However, from the present results, it was considered that replantation can be an effectual method when the condition of the avulsed primary tooth is suitable.


Effects of beta-lactam antibiotics on intestinal microflora and bile acid metabolism in rats.
Wistar male rats were treated for six days with broad spectrum beta-lactam antibiotics, latamoxef, and cefotaxime. On the seventh day, the number of fecal anaerobic microbes decreased, total fecal bile acids decreased, and bile acid pools increased. Secondary bile acids such as beta-hyocholic, hyodeoxycholic, lithocholic, and deoxycholic acids decreased in the feces while the primary bile acids, cholic, beta-muricholic, and chenodeoxycholic acids, became predominant. Coprostanol, a microbial metabolite of cholesterol, also disappeared from the feces during the treatment. The cecum enlarged to almost twice the size of that in control rats, whereas the liver weight was not significantly changed. After treatment was stopped, the number of fecal microbes returned to the initial counts within a week, but restoration of bile acid and cholesterol metabolism required at least three weeks.


Identification of the mutations in the prfB gene of Escherichia coli K12, which confer UGA suppressor activity.
By DNA sequencing and gene dissection, it has been revealed that Su+UGA#11, the mutant prfB of E. coli (Chang et al., 1990) has a double mutation compared with the wild-type LS653: one is a base substitution from T to C at the codon 63 and the other is from G to A at the codon 79. Both mutations cause amino acid substitution, Leu63----Phe63 (L63F) and Asp79----Gly79 (D79G), and are necessary to confer the efficient UGA suppressor activity.


Aberrant Rac pathway signalling in glioblastoma.
Glioblastoma is an aggressive and incurable form of brain cancer. Both mutation analysis in human glioblastoma and mouse modelling studies have shown that aberrant activation of the PI 3-kinase pathway is a central driver of glioblastoma malignancy. The small GTPase Rac is activated downstream of this pathway, mediating a subset of the effects of aberrant PI 3-kinase pathway activation. Here I discuss the current state of our knowledge on Rac activation mechanisms in glioblastoma. Current knowledge on roles for specific PI 3-kinase pathway responsive Rac guanine nucleotide exchange factors in glioblastoma is reviewed. Rac is best known for its role in promoting cell motility and invasion, but there is also evidence for roles in multiple other cellular processes with cancer relevance, including proliferation, differentiation, apoptosis, DNA damage responses, metabolism, angiogenesis and immunosuppression. I review what is known about the role of Rac in these processes in glioblastoma. Finally, I assess possible strategies to inhibit this pathway in glioblastoma through either direct inhibition of Rac or inhibition of upstream activators or downstream mediators of Rac signalling.


Cell division cycle in mammalian cells. VIII. Mapping of G1 into six segments using temperature-sensitive cell cycle mutants.
The G1 blocks in three temperature-sensitive (ts) Syrian hamster cell-cycle mutants have been mapped in relation to other G1 landmarks. Two mutants reported here, ts-559 and ts-694, show defective progression only in G1. When shifted from the permissive temperature of 33 degrees C to the non-permissive temperature of 39 degrees C, G1 cells of these two mutants show no further cell cycle progression, while cells in S, G2 and mitosis progress through the cell cycle but become blocked after entering G1. The two mutants complement each other, and also complement the previously reported mutant ts-550C with blocks in both G1 and G2 of the cell cycle. The locations of the G1 blocks in both ts-559 and ts-694 are before the hydroxyurea arrest point. The G1 ts point in ts-694 is prior to the isoleucine deprivation and serum starvation points, while the G1 block in ts-559 is after the serum starvation point but before the isoleucine block. Other G1 block points which have been reported are in mutants of different species and isolated in different laboratories, causing difficulties for relative positioning of the blocks in G1. The mutants for mapping in this study have been isolated from the same cell line. The G1 ts arrest points of ts-559 and ts-694, and that found in ts-550C, together with nutritional deprivations and metabolic inhibitors, provide seven reference points which divide G1 into six segments, each of which is bracketed by two adjacent points: mitosis, ts-694 block, serum starvation arrest point, ts-559 block, isoleucine deprivation arrest point, ts-550C block, hydroxyurea or excess-thymidine arrest segment.


Twice-daily compared with once-daily insulin glargine in people with Type 1 diabetes using meal-time insulin aspart.
To compare blood glucose control when using insulin glargine twice daily at breakfast- and dinner-times with insulin glargine once daily at dinner time, in unselected people with Type 1 diabetes using insulin aspart at meal-times. In this 8-week, two-way, cross-over study, 20 people with Type 1 diabetes were randomized to insulin glargine injection once daily at dinner-time or twice daily at breakfast- and dinner-times, both plus meal-time insulin aspart. Each 4-week treatment period concluded with a 24-h inpatient metabolic profile. Insulin doses, HbA1c, fructosamine concentration and pre-breakfast self-monitored blood glucose (SMBG) concentration did not differ between treatment periods. SMBG concentrations after breakfast, after lunch and before dinner were lower with twice-daily compared with once-daily dinner-time glargine [9.3 +/- 0.5 (+/- se) vs. 6.7 +/- 0.5 mmol/l, P = 0.003; 10.2 +/- 0.9 vs. 7.0 +/- 0.9 mmol/l, P = 0.024; 9.6 +/- 0.5 vs. 6.6 +/- 0.5 mmol/l, P = 0.001]. Mean 24-h SMBG concentration was lower with twice-daily glargine (7.1 +/- 0.5 vs. 8.8 +/- 0.5 mmol/l, P = 0.031). Within-day variability of SMBG concentration was lower with twice-daily glargine (sd 3.2 +/- 0.2 vs. 4.0 +/- 0.3 mmol/l, P = 0.044). Plasma free insulin concentration was higher in the afternoon with twice-daily glargine (21.9 +/- 1.4 vs. 16.1 +/- 1.3 mU/l, P = 0.009), but lower overnight (12.1 +/- 1.7 vs. 17.8 +/- 1.7 mU/l, P = 0.030), compared with once-daily injection. Plasma glucose concentration overnight was higher with twice-daily compared with once-daily glargine (mean 9.0 +/- 0.4 vs. 6.6 +/- 0.4 mmol/l, P = 0.001). Blood glucose concentration rises in the late afternoon in association with falling plasma insulin levels towards the end of the 24-h period after insulin glargine injection in some people with Type 1 diabetes using once-daily glargine at dinner-time plus a rapid-acting insulin analogue at meal-times. This is prevented by twice-daily injection of insulin glargine.


Enzymatic assessment of paraoxonase 1 activity on HDL subclasses: a practical zymogram method to assess HDL function.
We developed a practical method for analysis of PON-1 enzymatic activity in HDL subclasses. The assay uses 4-12% polyacrylamide gradient gels, phenylacetate as substrate coupled with densitometric phenol detection using 4-aminoantipyrine. The measurement PON-1 activity in situ across the HDL subclasses has a strong correlation with the kinetic microplate assay for total PON-1 activity, r=0.91, p<0.001. The same HDL-C level, healthy subjects (n=33) display a large difference in the ratio of PON-1 activity in small vs. large HDL. Since PON-1 activity is larger in HDL(3) we propose that this difference has a potent predictive value for clinical risk assessment and therapeutic choice. Our method also offers the advantage of assessing the distribution of up to six different HDL apolipoproteins in the same gel after transfer. We seek to further dissect the cause of a different distribution of PON-1 activity in HDL subclasses by employing this method that permits practical, inexpensive analysis of antioxidant function of HDL subclasses and has the potential for application in clinical chemistry and to shed some light on the importance of PON-1 distribution.


Clinical and demographic overlaps among immunologically mediated oral diseases: a challenge for clinicians.
This study sought to assess and compare retrospective demographic and clinical data of oral lesions of lichen planus, pemphigus vulgaris, and mucous membrane pemphigoid from the records of the Department of Oral Pathology and Surgery, School of Dentistry, Universidade Federal de Minas Gerais, Brazil, covering a period of 55 years. Out of 25,435 specimens, these immunologically-mediated diseases accounted for 301 (1.18%) cases, of which 250 (0.98%) were lichen planus, 27 (0.11%) were pemphigus vulgaris, and 24 (0.09%) were mucous membrane pemphigoid. Lichen planus presented mainly as white asymptomatic plaques on buccal mucosa. Pemphigus vulgaris was usually characterized by multiple symptomatic erithematous ulcers on the buccal mucosa. Painful ulcers and/or blisters on the gingiva were the most common presentation for mucous membrane pemphigoid. Desquamative gingivitis was noted for all 3 diseases, but mainly for mucous membrane pemphigoid. Overall, lesions were more frequent in white women >50 years. Oral manifestations of immunologically-mediated diseases are relatively rare, and the correct diagnosis can be a challenge for dentists as the lesions often share similar clinical and demographic features.


Molecular-biological analysis of acute lung injury (ALI) induced by heat exposure and/or intravenous administration of oleic acid.
The aim of this study was to molecular-biologically investigate the interaction between heat exposure and pulmonary fat embolization in regards to the development of acute lung injury (ALI). Ten-week-old Wistar male rats were divided into four groups: (1) oleic acid injected into caudal vein after heat exposure, (2) oleic acid injected without heat exposure, (3) soybean oil injected after heat exposure, and (4) soybean oil injected without heat exposure, and then mRNA expression of eight inflammatory mediators related to ALI/acute respiratory distress syndrome (ARDS) and heat shock protein 70 (Hsp70) in lung was determined 1h after the injection. mRNA expression of interleukin 1 beta (Il1b), tumor necrosis factor alpha (Tnfa), vascular endothelial growth factor A (Vegfa), transforming growth factor beta 1 (Tgfb1) and Hsp70 was significantly increased by heat exposure, while that of Il1b, interleukin 6 (Il6), Tnfa, macrophage inflammatory protein 2 (Mip2) and granulocyte macrophage-colony stimulating factor (Gm-csf) was significantly elevated by the injection of oleic acid. Moreover, the expressions of inflammatory cytokines and chemokines in lung almost paralleled their mRNA expressions. In particular, IL-1β expression was synergistically elevated by heat exposure followed by injection of oleic acid. Additionally, IL-6 expression tended to increase under the same conditions as well. It is likely that heat exposure itself injures lung tissue within a short time, and that more than two conditions which induce ALI/ARDS interact with each other synergistically, exacerbating the development of ALI/ARDS.


Beliefs about outcomes for mental disorders: a comparative study of primary health practitioners and psychiatrists in Singapore.
To compare responses to a mental health literacy survey assessing the likely outcome of three major mental disorders by primary health practitioners (OPD doctors and GPs) and by psychiatrists in Singapore. We used two vignettes of Major Depression and Schizophrenia developed in an Australian study. In addition, a third vignette of Mania was developed locally and included. The respondents were required to choose one of the set of prognostic options if the patients received or did not receive professional help, to rate the likely impact of the disorder, and to assess the likelihood of the patient being discriminated against. Psychiatrists' responses were obtained by surveying staff at Woodbridge Hospital, while the primary health practitioners were required to respond to a postal survey. The response rate for the psychiatrists was 70% (69/99), while the Primary health practitioners had an overall response rate of 38% (264/691). The response from OPDs being 51% (77/151) and that of the GPs being 35% (189/540). There was evidence of disorder specificity, with schizophrenia judged as having the worst outcome and depression the best outcome in response to treatment. There was also evidence of group specificity, with the psychiatrists most optimistic and the OPD doctors least optimistic about the outcome following professional intervention. The majority of both the primary health practitioners and the psychiatrists judged that patients would be discriminated against, more so for schizophrenia and mania than for depression. Compared to the OPD doctors, a lower percentage of GPs felt that the patients would be discriminated against. Primary health practitioners in Singapore hold more negative views than Singapore psychiatrists about the outcome of professional intervention for three major psychiatric disorders. This finding has implications for education and training for primary health practitioners as well as for treatment of psychiatric patients in the primary health setting.


Synthesis of [11C]palmitic acid for PET imaging using a single molecular sieve 13X cartridge for reagent trapping, radiolabeling and selective purification.
Radiolabeled fatty acids are valuable metabolic tracers for PET imaging. Carbon-11 is widely used in clinical PET studies due to the prevalence of facile techniques enabling the incorporation of [(11)C]CO2 and [(11)C]CH3 into molecules and a short half-life (20.4 min) that translates into low patient dose. However, the short half-life considerably limits the time for radiosynthesis. Furthermore, the majority of the syntheses of [(11)C]palmitic acid in common use employ high starting [(11)C]CO2 activities and/or expensive equipment. [(11)C]CO2 was trapped with greater than 99.99% efficiency by a three stage cartridge packed with molecular sieve 13X, 100-120 mesh. The labeling of n-pentadecylmagnesium bromide took place in 5 min in the cartridge, and the [(11)C]palmitic acid product was selectively eluted in ethanol following alkaline and acidic washes of the column. The system reliably produced more than 925 MBq (25 mCi) of [(11)C]palmitic acid suitable for human use from 7.4 GBq (200 mCi) of [(11)C]CO2 in 8 min from end-of-bombardment. We have exploited the properties of the inexpensive molecular sieve 13X to develop a miniature, disposable and leak tight "gas capture" system for the rapid labeling and purification of [(11)C]fatty acids in good yield and >99% radiochemical purity. The rapidity of the synthesis and purification allows small [(11)C]CO2 starting activities to be used, and with no requirement for expensive synthesis equipment or facilities, the system can be implemented in any radiopharmaceutical center.


Assembly, intracellular processing, and expression at the cell surface of the human alpha beta T cell receptor/CD3 complex. Function of the CD3-zeta chain.
The TCR/CD3 complex is a multimeric protein complex composed of a minimum of seven transmembrane chains (TCR alpha beta-CD3 gamma delta epsilon zeta 2). Whereas earlier studies have demonstrated that both the TCR-alpha and -beta chains are required for the cell surface expression of the TCR/CD3 complex, the role of the CD3 chains for the TCR/CD3 expression have not been experimentally addressed in human T cells. In this study the function of the CD3-zeta chain for the assembly, intracellular processing, and expression of the TCR/CD3 complex in the human leukemic T cell line Jurkat was investigated. The results indicate that: 1) CD3-zeta is required for the cell surface expression of the TCR/CD3 complex; 2) the pentameric form (TCR alpha beta-CD3 gamma delta epsilon) of the TCR/CD3 complex and single TCR chains associated with CD3 (TCR alpha-CD3 gamma delta epsilon and TCR beta-CD3 gamma delta epsilon) are produced in the endoplasmic reticulum in the absence of CD3-zeta; 3) the CD3-zeta does not associate with TCR alpha-CD3 gamma delta epsilon or TCR beta-CD3 gamma delta epsilon complexes; 4) CD3-zeta associate with the pentameric form of the TCR/CD3 complex in the endoplasmic reticulum to form the heptameric complex (TCR alpha beta-CD3 gamma delta epsilon----TCR alpha beta-CD3 gamma delta epsilon 2); and 5) CD3-zeta is required for the export of the TCR/CD3 complex from the endoplasmic reticulum to the Golgi apparatus for subsequent processing.


Weight management in primary care: how can it be made more effective?
Obesity is often difficult to tackle in primary care. Pressure of time in the consultation, a lack of appropriately-trained primary care staff, a shortage of community dietitians or nutritionists, the potentially enormous caseload, language or cultural barriers and the sheer intractability of patients' eating habits, exercise behaviour and their clinical condition, all conspire to make general practitioners, other team members and often the patients themselves lose heart and stop even trying. However, there are ways of overcoming these difficulties. Examples of changes that evidence suggests are able to support and enhance basic one-to-one interventions in general practice include: improved clinical guidelines; better training of primary care staff; at-risk patient registers; smarter database search tools; new quality incentives; closer working with dietitians, counsellors and pharmacists; more hospital outreach clinics; designated general practitioner specialists and practice clustering; expanded exercise referral schemes and links with leisure providers; subsidised referral to commercial slimming groups; better use of patient groups and voluntary and community workers. The present paper describes a proposed 'triple-tier' pathway for weight management incorporating most of the elements mentioned earlier. With a more joined-up and creative approach to the development and organisation of primary care, more comprehensive training and workforce planning, and better integration with social care, voluntary groups and the commercial sector, weight management in general practice has the potential to be much more effective.


Y-chromosomal STR haplotypes in a Belgian population sample and identification of a micro-variant with a flanking site mutation at DYS19.
Allele frequencies and haplotypes for 12 Y-chromosomal STR loci included in the Powerplex System (Promega, Madison, USA) were determined in a sample of 113 unrelated males of Belgian origin. Ninety-nine different haplotypes were observed with an overall haplotype diversity of 0.997.


Molecular characterization of Aspergilli isolated from outdoor air.
Ubiquitous airborne conidia of the genus Aspergillus are responsible for a diverse group of human disorders from allergy to life treating invasive aspergillosis and mycotoxicoses. The aim of this study was to determine the population structure of Aspergillus isolated from outdoor air in Tehran by comparing the nucleotide sequences of ITS region and the PCR-RFLP molecular method. Internal transcribed spacer domains of 47 Aspergillus spp. were amplified and sequenced and PCR products were digested individually with restriction enzymes TaqI and EcoRI. For all species the PCR reaction produced a fragment of approximately 600bp in length. All of the nucleotide sequences were highly similar with the corresponding reference sequences registered at the gene bank. The all isolates displayed same banding pattern on the basis EcoR1 cleavage. While Taq1 enzyme profiling provided 5 different banding pattern. The results show that the A. niger section has the highest frequency with 27 isolates (57.4%). Of these, 23 isolates (48.9%) belonged to the A. niger complex and 4 isolates (8.5%) to the A. aculeatus complex. The A. flavus complex was also placed in the next ranking with 9 isolates (19.1%). These results strongly support the need for using molecular markers as an auxiliary tool in differentiating Aspergillus species.


A buyer's guide to the innovation bazaar.
Companies seeking new ideas or product concepts from outside sources may find the "innovation bazaar," with its wide array of choices and methods of acquiring them, a confusing, chaotic place. Nambisan and Sawhney have crafted a conceptual guide for managers who understand the importance of going outside their firms for innovation but are uncertain about how to do it. The authors' "external sourcing continuum" shows at a glance how shopping for, say, raw ideas compares with shopping for market-ready products in terms of cost, risk, multiplicity of options, and speed of commercialization. Raw ideas, whether acquired directly from the inventor or through a patent broker, licensing agent, or some other intermediary, tend to be low cost but high risk and take a long time to bring to market. Market-ready products, often acquired as stand-alone businesses through a venture capitalist or business incubator, are more expensive and narrow one's choices, but they can be launched quickly and with less risk. Between these two approaches lies a third, facilitated by the "innovation capitalist." This new kind of intermediary provides client companies with access to a broad range of innovative product or technology ideas that are nearly market ready, thereby mitigating early-stage risks and lowering the time to market without significantly increasing acquisition costs. The authors compare the advantages and disadvantages of using intermediaries associated with the three approaches and provide a checklist of factors to consider when placing your company on the external sourcing continuum. If you've been oriented toward one end of the continuum or the other, you can increase your options and your flexibility by expanding into the middle.


[Familial occurrence of central diabetes insipidus].
Two cases are presented of familial diabetes insipidus with analysis of pedigree involving four generations. The analysis of diabetes insipidus occurrence in this family indicates the autosomal dominant type of inheritance.


Role of prophylaxis in vesicoureteral reflux.
Traditional management of vesicoureteral reflux focuses on preventing renal complications associated with ascending urinary tract infection by either providing continuous antibiotic prophylaxis to sterilize the urine and thus prevent recurrent infection, or abolishing reflux via surgical intervention. This review will consider the rationale for antibiotic prophylaxis in light of contemporary data regarding the natural history of vesicoureteral reflux, urinary tract infection and renal scarring, as well as the efficacy of various treatment strategies. Recent studies have shown that in grades I-IV vesicoureteral reflux, open surgical intervention compared with antibiotic prophylaxis is no better at preventing renal complications. Endoscopic subureteral injection of biomaterials has been proposed as a cost-effective, minimally invasive alternative to surgical or medical intervention; however, given the variety of materials and techniques, the literature has yet to achieve consensus on its efficacy. The first study to compare antibiotic prophylaxis with increased surveillance and prompt treatment of urinary tract infection shows no difference between the two approaches in low-grade (grades I-II) vesicoureteral reflux. Recent studies have challenged the traditional paradigm of aggressive vesicoureteral reflux management with surgery or antibiotic prophylaxis. In light of these findings, pediatric urologists will need to reexamine treatment modalities for vesicoureteral reflux.


Studies on the mechanism of electron trasport in the bc1-segment of the respiratory chain in yeast. III. Isolation and characterization of an antimycin resistant mutant ANT 8 in Schizosaccharomyces pombe.
1. A mutant (ANT 8) of Schizosaccharomyces pombe which shows resistance to antimycin both in vivo and in vitro is characterized biochemically and genetically. 2. In crosses of ANT 8 with auxotrophic strains, resistance to antimycin segregates 2:2 indicating that resistance is conferred by a single nuclear gene. Diploids heterozygous for the resistance gene, however, show segregation of the resistance and sensitivity during mitosis. Possible reasons for this segregation are discussed. 3. Compared with the wild type, the NADH oxidase of ANT 8 requires 13 times as much antimycin for 95% inhibition. After addition of ubiquinone-3, electron transport which is less sensitive to antimycin is found only in the mutant. 4. The resistance of the mutant ANT 8 si due to the much weaker binding of antimycin to mitochondria. As in the wild type, two antimycin binding sites can be separated by binding studies. From the inhibition curves it is evident that binding of antimycin to oxidized mitochondrial particles does not correspond with its inhibitory effect on the partly reduced enzyme in kinetic studies. 5. The peak of the b-cytochrome absorbing at 560.2 nm at 77 degrees K in the wild type is shifted to 561 nm in the mutant. 6. A special preparation method for mutant mitochondrial particles is described, yielding highly active enzymes and CO-insensitive cytochromes. 7. The results are discussed with reference to the components in our model of the respiratory chain, which may be responsible for this type of resistance.


Helminth infections are associated with protection from cerebral malaria and increased nitrogen derivatives concentrations in Thailand.
Following a study showing an association between Ascaris and protection from cerebral malaria, we hypothesized helminths may have induced protection through immunoglobulin E (IgE) and the CD23/NO pathway. We compared the prevalence of helminth infections in 67 cerebral malaria patients and 217 hyperparasitemic controls with no complications. For 24 cerebral malaria cases and 56 controls, we compared reactive nitrogen intermediates (RNI) concentrations and their correlations to total IgE and sCD23 concentrations in helminth-infected and noninfected patients. We observed a dose-dependent association between helminth infections and protection from cerebral malaria (adjusted odds ratio [OR] = 0.36, 95% CI = 0.19-0.7, P = 0.002, linear trend P = 0.0007). Helminth-infected controls had higher RNI concentrations than those without helminths: 72 OD +/- 19 SD and 57 OD +/- 20 SD, respectively (P = 0.006). Logistic regression, including interaction terms between RNI and sCD23, showed that an increase of RNI could be both protective and pathogenic depending on the concentration of sCD23. Helminths increasing both the CD23 receptor and its ligand may have a role in the establishment of malaria tolerance through the CD23/NO pathway.


Assessing African medicinal plants for efficacy and safety: agricultural and storage practices.
The paper reviews an important but little researched area of ethnopharmacology, namely, the effect of cultivation and post-harvest storage practices on levels of biological activity in traditionally used medicinal plants. Changes in COX-1 inhibition and antibacterial activity, for example, occur at the onset of senescence and, in some species, are influenced by plant age. Plants in cultivation were also shown to have reduced anthelmintic and antibacterial activity while hypoxoside production was affected by nitrogen and phosphorous. Irrigation treatments, however, increased anthelmintic activity and, therefore, could be applied to medicinal plants cultivated in low rainfall areas. Pesticides have also been implicated in the regulation of plant growth and secondary metabolite production in cultivated medicinal plants, but residue levels have, so far, not been monitored. Post harvest storage of medicinal plants has been poorly researched in southern Africa. Available data indicates that antibacterial and anti-inflammatory activity changes following storage and is both species and temperature dependent. Similar trends were noted for materials that had been rapidly aged. Fatty acids with antibacterial activity are stable in dry specimens and, as such, may account for the fact that activity is unaffected by storage in certain instances.


[The insertion of the nurse into the work market place--some considerations].
The present study makes some considerations on nurse's insertion in work field. It rises some data on the increase of employment in health sector, as well as seeks to identify some determinants of this increasment. It brings up some informations on nurse's manpower, the absorption of nursing professional ad the ability of generating employment. Finally, the study identify some trends perceived in the absorption of the professional nurse.