Analysis of the variation in use-dependent inactivation of high-threshold tetrodotoxin-resistant sodium currents recorded from rat sensory neurons.
This study addressed variation in the use-dependent inactivation (UDI) of high-threshold tetrodotoxin-resistant Na+ currents (TTX-R currents) and action potential firing behavior among acutely isolated rat dorsal root ganglion (DRG) cells. UDI was quantified as the percent decrease in current amplitude caused by increasing the current activation rate from 0.1-1.0 Hz for 20 s. TTX-R current UDI varied from 6% to 66% among 122 DRG cells examined, suggesting the existence of two or more levels of UDI. The voltage-dependency of the TTX-R currents was consistent with Na(V)1.8, regardless of UDI. However, TTX-R currents with more UDI had a more negative voltage-dependency of inactivation, a greater tendency to enter slow inactivation, and a slower recovery rate from slow inactivation, compared with those with less UDI. TTX-R currents with more UDI ran down faster than those with less UDI. However, UDI itself changed little over time, regardless of the initial UDI level observed in a particular DRG cell. Together, these two observations suggest that individual DRG cells did not express mixtures of TTX-R channels that varied regarding UDI. TTX-R current UDI was correlated with expression of a low-threshold A-current and whole-cell capacitance, suggesting that it varied among different nociceptor types. Whole-cell inward currents (WCI-currents), recorded without channel blockers, also exhibited UDI. WCI-current UDI varied similarly to TTX-R current UDI in magnitude, and relative to whole-cell capacitance and A-current expression, suggesting that the WCI-currents were carried predominantly by TTX-R channels. DRG cells with more WCI-current UDI exhibited a greater decrease in action potential amplitude and number, and a greater increase in action potential threshold over seven ramp depolarizations, compared with DRG cells with less WCI-current UDI. Variation in UDI of Na(V)1.8 channels expressed by different nociceptor types could contribute to shaping their individual firing patterns in response to noxious stimuli.


Metagenomic Evaluation of Bacteria from Voles.
Voles (Arvicolinae, Rodentia) are known carriers of zoonotic bacteria such as Bartonella spp. and Francisella tularensis. However, apart from F. tularensis, the bacterial microbiome of voles has not previously been determined in Finland and rarely elsewhere. Therefore, we studied liver samples from 61 voles using 16S ribosomal RNA gene PCR analysis, followed by Sanger sequencing. Twenty-three of these samples were also studied with tag-encoded pyrosequencing. The samples originated from 21 field voles (Microtus agrestis), 37 tundra voles (Microtus oeconomus), and 3 bank voles (Myodes glareolus). With the more conventional 16S rDNA PCR analysis, 90 (33%) of the recovered 269 sequence types could be identified to genus level, including Bartonella, Francisella, Mycoplasma, Anaplasma, and Acinetobacter in 31, 15, 9, 9, and 9 sequences, respectively. Seventy-five (28%) matched best with sequences of uncultured bacteria, of which 40/75 could be classified to the order Clostridiales and, more specifically, to families Lachnospiraceae and Ruminococcaceae. Pyrosequencing from 23 samples revealed comparable and similar results: clinically relevant bacterial families such as Mycoplasmataceae, Bartonellaceae, Anaplasmataceae, and Francisellaceae were recognized. These analyses revealed significant bacterial diversity in vole livers, consisting of distinct and constant sequence patterns reflecting bacteria found in the intestinal gut, but including some known zoonotic pathogens as well. The molecular bacterial sequence types determined with the two different techniques shared major similarities and verified remarkable congruency between the methods.


A review of modern sample-preparation techniques for the extraction and analysis of medicinal plants.
Sample preparation is the crucial first step in the analysis of herbs. In recent years there has been increasing interest worldwide in the use of alternative/herbal medicine for the prevention and treatment of various illnesses. Currently, however, quality-related problems (lack of consistency, safety, and efficacy) seem to be overshadowing the potential genuine health benefits of various herbal products. Thus, the development of "modern" sample-preparation techniques with significant advantages over conventional methods for the extraction and analysis of medicinal plants is likely to play an important role in the overall effort of ensuring and providing high-quality herbal products to consumers worldwide. In this article, recent developments and applications of modern sample-preparation techniques for the extraction, clean-up, and concentration of analytes from medicinal plants or herbal materials are reviewed. These modern techniques include solid-phase microextraction, supercritical-fluid extraction, pressurized-liquid extraction, microwave-assisted extraction, solid-phase extraction, and surfactant-mediated extraction.


Design and development of amplifiable broad-host-range cloning vectors: analysis of the vir region of Agrobacterium tumefaciens plasmid pTiC58.
The construction of a set of new plasmids that are suitable as general cloning vectors in Escherichia coli and Agrobacterium tumefaciens is described. Plasmid pUCD2 is amplifiable in E. coli, replicates in a wide range of gram-negative hosts and contains a number of useful restriction endonuclear cleavage sites and antibiotic resistance genes. This includes unique sites for KpnI, SacI, SacII, PstI, ClaI, SalI, EcoRV, and PvuII and the genes for resistance to kanamycin, tetracycline, ampicillin, and spectinomycin/streptomycin. Derivatives of pUCD2 include pUCD4, which has a unique XbaI site and the cosmid pUCD5, which also contains a unique EcoRI site. Two smaller plasmids pUCD9P and pUCD9X, contain many of the same unique sites as pUCD2 and pUCD4, but carry only the pBR322 replication origin and therefore do not display the extensive host-range of pSa. These plasmids were used to isolate and manipulate fragments of the A. tumefaciens pTiC58 plasmid in both E. coli and A. tumefaciens. Fragments from the virulence (vir) region of pTiC58 inserted immediately upstream of the spectinomycin resistance gene of pUCD2 resulted in spectinomycin resistance levels that varied greatly depending on the particular fragment and its orientation of insertion. Using this property we find that a major portion of the vir region of pTiC58 is transcribed in A. tumefaciens and E. coli from left to right toward the T region.


Automatic segmentation for adaptive planning in nasopharyngeal carcinoma IMRT: Time, geometrical, and dosimetric analysis.
The aim of this study was to quantify the geometrical differences between manual contours and autocontours, the dosimetric impacts, and the time gain of using autosegmentation in adaptive nasopharyngeal carcinoma (NPC) intensity-modulated radiotherapy (IMRT) for a commercial system. A total of 20 consecutive Stages I to III NPC patients who had undergone adaptive radiation therapy (ART) re planning for IMRT treatment were retrospectively studied. Manually delineated organs at risks (OARs) on the replanning computed tomography (CT) were compared with the autocontours generated by VelocityAI using deformable registration from the original planning CT. Dice similarity coefficients and distance-to-agreements (DTAs) were used to quantify their geometric differences. IMRT test plans were generated with the assistance of RapidPlan based on the autocontours of OARs and manually segmented target volumes. The dose distributions were applied on the manually delineated OARs, their dose volume histograms and dose constraints compliances were analyzed. Times spent on target, OAR contouring, and IMRT replanning were recorded, and the total time of replanning using manual contouring and autocontouring were compared. The averaged mean DTA of all structures included in the study were less than 2 mm, and 90% of the patients fulfilled the mean distance agreement tolerance recommended by AAPM 132.1 The averaged maximum DTA for brainstem, cord, optic chiasm, and optic nerves were all less than 4 mm, whereas temporal lobes and parotids have larger average maximum DTA of 4.7 mm and 6.8 mm, respectively. It was found that large contour discrepancies in temporal lobes and parotids were often associated with large magnitude of deformation (warp distance) in image registrations. The resultant maximum dose of manually segmented brainstem, cord, and temporal lobe and the median dose of manually segmented parotids were found to be statistically higher than those to their autocontoured counter parts in test plans. Dose constraints of the manually segmented OARs in test plans were only met in 15% of the cases. The average time of manual contouring and autocontouring were 108 and 10 minutes, respectively (p < 0.001). More than 30% of the total replanning time would be spent in manual OAR contouring. Manual OAR delineation takes up a significant portion of time spent in ART replanning and OAR autocontouring could considerably enhance ART workflow efficiency. Geometrical discrepancies between auto- and manual contours in head and neck OARs were comparable to typical interobserver variation suggested in various literatures; however, some of the corresponding dosimetric differences were substantial, making it essential to carefully review the autocontours.


Endoparasitic mites of the genus Endotrombicula Ewing, 1931 (Acari: Prostigmata: Parasitengona: Trombiculidae) from African and Madagascan anurans, with description of a new species.
A sample of over 6,000 specimens of frogs belonging to about 120 species of all families occurring in West Africa and Madagascar were screened for parasitic mites. Three species of Endotrombicula Ewing, 1931 were found in representatives of two African and two Madagascan frog families. All Trombiculidae found in African frogs belonged to Endotrombicula pillersi (Sambon, 1928), whereas in Madagascar E. madagascariensis (Sambon, 1928) and E. ptychadenae sp. n. were sampled. These three species are described, data about their parasitic associations are provided, and their zoogeographical distribution is discussed. Only those frog species that spend a considerable time in terrestrial ground habitats were parasitized; neither arboreal nor strictly aquatic frogs were infected. The geographic distribution of Endotrombicula, restricted to Africa, the Arabian Peninsula and Madagascar, suggests that these mites invaded Madagascar from the African continent. This is supported by the observation that the ancestors of Ptychadena mascareniensis (Duméril et Bibron) (Ptychadenidae), the host of E. ptychadenae, colonized Madagascar from the African continent quite recently, possibly accompanied by its Endotrombicula parasites.


Alpha- and beta-mannosidoses.
Clinical, pathological and biochemical findings in the mannosidoses are described. Family studies showed granulocyte-rich white cell fractions to be the tissue of choice for carrier detection in beta-mannosidosis. Metabolic labelling studies using [3H] mannose demonstrated accumulation of Man beta 1-4GlcNAc in cultured skin fibroblasts from a patient with this condition. Alternative methods of egress from lysosomes were suggested for this compound by its secretion into culture medium and apparent reduction of storage with time in cultures. beta-mannosidase deficient goats are not thought to be a true animal model of the human condition, as although they showed a similar enzyme deficiency, the clinical presentation is much more severe and the major storage material (Man beta 1-4GlcNAc beta 1-4GlcNAc) is different.


Synthesis of RNA oligomer using 9-fluorenylmethoxycarbonyl (Fmoc) group for 5'-hydroxyl protection.
An approach using a new combination of protecting groups in RNA oligomer synthesis is proposed, in which 5'-hydroxyl group of ribose moiety is temporarily protected with the alkaline labile 9-fluorenylmethoxycarbonyl (Fmoc) group and the 2'-hydroxyl group is protected with the acid labile 1-ethoxyethyl (EE) group. The adoption of this method presented great selectivity in removing the 5'-hydroxyl protecting group and facilitated the RNA oligomer synthesis. A RNA pentamer was synthesized by the phosphotriester method in solution.


Effect of lipid load and carbon tetrachloride on the fatty acid release from dog adipose tissue isolated in vivo.
Under the influence of lipid emulsion (LipofundinR) administered into the systemic circulation of dogs, fatty acid release of adipose tissue isolated in vivo increased from 0.51 +/- 0.096 muEq/100 g/min to 3.18 +/- 0.48 muEq/100 g/min. If the lipid emulsion had been injected into the artery of adipose tissue, the fatty acid release was elevated from 0.34 +/- 0.053 muEq/100 g/min to 1.67 +/- 0.25 muEq/100 g/min. The effect could be observed in denervated adipose tissue too and in this case the fatty acid release increased from 0.40 +/- 0.053 muEq/100 g/min to 1.16 +/- 0.14 muEq/100 g/min. If CCl4 dissolved in lipid emulsion was injected into the systemic circulation, there was no significant change and a value of 0.45 +/- 0.063 muEq/100 g/min was measured. However, on administering the CCl4 dissolved in LipofundinR into the artery of adipose tissue, the fatty acid release rose from 0.34 +/- 0.053 to 1.01 +/- 0.18 muEq/100 g/min. The same manipulation on denervated adipose tissue elevated the fatty acid release from 0.40 +/- 0.053 to 0.96 +/- 0.16 muEq/100 g/min.


Noninvasive venous testing in the diagnosis of pulmonary embolism: the impact on decisionmaking.
To characterize the use and utility of lower extremity noninvasive venous testing (NIVT) in the diagnosis of pulmonary embolism (PE). The study is a retrospective case series of consecutive patients in whom PE was suspected who were referred to a large, urban tertiary care center for NIVT. The main outcome measures of the study were the rate of positive results of NIVT, the amount of new information provided by NIVT, and the frequency of management changes that were attributable to NIVT. Forty-one of 450 patients (9%) had deep venous thrombosis (DVT) by NIVT. The prevalence of DVT by NIVT among patients not evaluated by ventilation/perfusion (V/Q) scanning was 8%. The prevalence of DVT by NIVT among patients with a high-probability V/Q scan result before NIVT was 39%, but no management decisions in this group were based on a positive NIVT result and only two decisions were based on negative NIVT results. The prevalence of DVT according to NIVT among patients who had a negative "diagnostic" (low, or very low probability, or normal) result of V/Q scan before NIVT was 2%. The overall frequency of management changes attributed to NIVT was only 2.5%. In the remaining 97% of patients, management was determined by the result of V/Q scanning or of subsequent pulmonary arteriography. In patients in whom PE is suspected, results of NIVT are usually negative for acute DVT. Management decisions are almost always based on V/Q scan or results of pulmonary arteriography and not on NIVT. The utility of NIVT to identify DVT in these patients appears limited, and a more selective approach to its application for the diagnosis of PE should be considered.


Skin mini-erosion technique for monitoring metabolites in interstitial fluid: its feasibility demonstrated by OGTT results in diabetic and non-diabetic subjects.
The aim of this study was to evaluate a new interstitial fluid (IF) sampling technique and its application in diabetology. IF and venous whole blood were sampled serially during an oral glucose tolerance test (OGTT) on days 1 and 3 after formation of a mini-erosion in eight non-diabetic controls and eight Type 2 diabetic subjects. Glucose, lactate, glycerol, 3-hydroxybutyrate and insulin were assayed in IF and plasma. With solitary exceptions, the IF sample volumes were in excess of those required for measurement of all five substances. However, mean IF sampling rates differed significantly both between the non-diabetic and diabetic groups and between days 1 and 3 (p < 0.001 in all cases). In both groups, the OGTT curves of glucose, glycerol and 3-hydroxybutyrate were similar to the respective curves for plasma, whereas for lactate concentrations they were markedly greater in IF than in plasma (40% and 93% in the control group and 57% and 100% greater in the diabetic group on days 1 and 3, respectively). The reverse was true of insulin concentrations in the non-diabetic group, which were 57% and 74% lower in IF than in plasma on days 1 and 3, respectively. In the non-diabetic group, the baseline (pre-OGTT) insulin level in IF increased from 49 +/- 22% (SD) of that in plasma on day 1 to 74 +/- 19% of that in plasma on day 3 (p=0.005). Sampling site re-epithelialization was rapid. In conclusion, the feasibility of transdermal sampling of IF via a skin mini-erosion has been demonstrated in both diabetic and non-diabetic subjects.


Strategies to utilize marker-quantitative trait loci associations.
Marker-assisted selection holds promise because genetic markers provide completely heritable traits than can be measured at any age in either sex and that are potentially correlated with traits of economic value. Theoretical and simulation studies show that the advantage of using marker-assisted selection can be substantial, particularly when marker information is used, because normal selection is less effective, for example, for sex-limited or carcass traits. Assessment of the available information and its most effective use is difficult, but approaches such as crossvalidation may help in this respect. Marker systems are now becoming available that allow the high density of markers required for close associations between marker loci and trait loci. Emerging technologies could allow large numbers of polymorphic sites to be identified, practically guaranteeing that markers will be available that are in complete association with any trait locus. Identifying which polymorphism out of many that is associated with any trait will remain problematic, but multiple-locus disequilibrium measures may allow performance to be associated with unique marker haplotypes. This type of approach, combined with cheap and high density markers, could allow a move from selection based on a combination of "infinitesimal" effects plus individual loci to effective total genomic selection. In such a unified model, each region of the genome would be given its appropriate weight in a breeding program. However, the collection of good quality trait information will remain central to the use of these technologies for the foreseeable future.


[Endoscopic exploration of the small intestine].
The development of Japanese enteroscopes, mot of which are still prototypes, has made it possible to explore the entire small bowel. These instruments are either introduced into the jejunum like an ordinary endoscope, or pushed along a guidewire, or weighted with mercury, like a Miller-Abbott probe. A simplified study of the adult malabsorption syndrome can be performed by proximal jejunoscopy, usually carried out in Europe with a paediatric colonoscope. Perendoscopic forceps biopsy of the jejunum can replace probe sampling and is well suited to measurements of disaccharidase activity. In cases with "blind loop syndrome", bacteriological examination of the intestinal juice, endoscopically collected in sterile conditions, usefully supplements the visual and bioptic information. Since several investigations can be performed during one single enteroscopic examination, this method will be increasingly used in the study of small bowel diseases.


Epidemiology of infant deaths due to congenital malformations: Italy 1958-1981.
The spatial and temporal distribution of infant death rates attributed to the single cause of congenital malformation (DCM) in Italy was studied for the period 1958-1981. Variation of DCM rates in this period was investigated for the whole country, for the three main geographical areas of the North, Center, and South, and for each of the 18 Italian regions (there are 20 administrative regions, but in this study Abruzzo were pooled with Molise and the Aosta Valley with neighbouring Piedmont). It was found that DCM decreases significantly with time. The decrease is considerable in the North and the Center of the country, whereas in the South temporal variation is minimal. There is a marked decrease of the variance of the DCM rates between regions with later years, possibly indicating a tendency to equalization of environmental effects in a large part of the peninsula.


Analysis of p53 gene polymorphisms and protein over-expression in patients with breast cancer.
p53 polymorphic variants play an important role in the determination of tumor phenotype and characteristics in breast cancer. In this study, we examined three common polymorphisms in p53 gene and their haplotype combinations to assess their potential association with inherited predisposition to breast cancer development, in relations with the protein over-expression and patients' demographic data. A total of 99 patients with breast cancer and 107 age-matched healthy controls were included in the study. Genotypes were determined using PCR-RFLP and DNA sequencing techniques. Evaluation of p53 protein over-expression was also examined by immunohistochemistry. Among three polymorphisms, increased codon 72 Pro allele frequency (p = 0.0067) and the presence of Pro allele were found to be significantly associated with breast cancer (p = 0.013). A significant risk was also found in subjects with combinations of specific haplotypes and genotypes. Most of breast cancer women especially younger than 50 years carry at least one p53 polymorphism (p = 0.001). There was no any association between these three p53 polymorphisms and the protein over-expression, separately or in interaction, with breast cancer. In conclusion, presence of proline allele at codon 72 alone, and its special combinations with other two polymorphisms appear to be a significant risk factor for breast cancer. Determination of well-known p53 polymorphisms might be a good predictor for breast cancer development especially in women younger than 50 years.


Adverse life events and psychopathology and prosocial behavior in late adolescence: testing the timing, specificity, accumulation, gradient, and moderation of contextual risk.
To explore the timing, specificity, accumulation, gradient, and moderation of contextual risk in psychopathology and prosocial behavior in late adolescence. In 2006, three hundred eighty-one 16- to 18-year-olds in Britain reported on the number and type of adverse life events experienced in the past month, when they were age 15, and when they were age 10, and on their concurrent psychopathology and prosocial behavior. They also undertook a reasoning ability test. Control variables were sex, age, and mothers' and fathers' educational attainment when participants were age 10 years. Although the number of adverse life events irrespective of their timing was associated with emotional and behavioral problems, the number of proximal adverse life events experienced was associated with psychopathology over and above the association of contextual risk in late childhood and in middle adolescence with psychopathology. The cumulative risk model was more parsimonious than the specific risk model. The relationship between proximal cumulative adversity and psychopathology was monotonic, and reasoning ability buffered the association of proximal cumulative adversity with psychopathology, mainly because it moderated the association of proximal cumulative adversity with hyperactivity. This study highlighted the importance of exploring risk accumulation rather than specificity in explaining psychopathology and showed that the number of adverse life events experienced has a nonmultiplicative association with psychopathology.


Controversies in the evolutionary social sciences: a guide for the perplexed.
It is 25 years since modern evolutionary ideas were first applied extensively to human behavior, jump-starting a field of study once known as 'sociobiology'. Over the years, distinct styles of evolutionary analysis have emerged within the social sciences. Although there is considerable complementarity between approaches that emphasize the study of psychological mechanisms and those that focus on adaptive fit to environments, there are also substantial theoretical and methodological differences. These differences have generated a recurrent debate that is now exacerbated by growing popular media attention to evolutionary human behavioral studies. Here, we provide a guide to current controversies surrounding evolutionary studies of human social behavior, emphasizing theoretical and methodological issues. We conclude that a greater use of formal models, measures of current fitness costs and benefits, and attention to adaptive tradeoffs, will enhance the power and reliability of evolutionary analyses of human social behavior.


[Does aircraft noise affect health?].
A field study of the Institute of Behavioral Sciences at the Federal Institute of Technology, Zürich, has verified that aircraft noise effects severe annoyance and inconvenience of airport area residents. This causes irritation, which manifests itself in numerous negative intentions, attitudes and opinions, in actions and meaures and even in psychosomatic disorders. Moreover noise exerts influence on vegetative reactions of persons performing different activities, mostly if they have to concentrate on a task.


Barfly: sculpting membranes at the Drosophila neuromuscular junction.
The ability of a cell to change the shape of its membranes is intrinsic to many cellular functions. Proteins that can alter or recognize curved membrane structures and those that can act to recruit other proteins which stabilize the membrane curvature are likely to be essential in cell functions. The BAR (Bin, amphiphysin, RVS167 homology) domain is a protein domain that can either induce lipidic membranes to curve or can sense curved membranes. BAR domains are found in several proteins at neuronal synapses. We will review BAR domain structure and the role that BAR domain containing proteins play in regulating the morphology and function of the Drosophila neuromuscular junction. In flies the BAR domain containing proteins, endophilin and syndapin affect synaptic vesicle endocytosis, whereas CIP4, dRich, nervous wreck and syndapin affect synaptic morphology. We will review the growing evidence implicating mutations in BAR domain containing proteins being the cause of human pathologies.


Cholinergic blockade with pirenzepine induces dose-related reduction in glucose and insulin responses to a mixed meal in normal subjects and non-insulin dependent diabetics.
The aim of the study was to assess the effects of cholinergic blockade with pirenzepine on glucose and insulin responses to a mixed meal in normal subjects and patients with non-insulin dependent diabetes (NIDDM). Further, to assess in normal subjects the relative importance of nocturnal GH suppression by pirenzepine. Placebo, 100 or 200 mg pirenzepine were given to the normal subjects 1 hour before a standard mixed meal. The effects of placebo or 200 mg pirenzepine at night on nocturnal GH secretion and subsequent breakfast carbohydrate tolerance were also studied. NIDDMs were given placebo or 200 mg pirenzepine before the meal. We studied six healthy male volunteers (ages 20-22, body mass indices 20.3-23.3) and ten NIDDMs (eight men, ages 42-74); five obese (BMI 25.5-31.8) and five non-obese (BMI 21.2-24.8). Serial plasma glucose and insulin concentrations were measured for 3 hours after a standard mixed meal. Acute pretreatment of normal male volunteers with pirenzepine produced a dose-related improvement in carbohydrate tolerance. Peak post-prandial plasma glucose levels were delayed and significantly reduced following 200 mg orally (6.5 +/- 0.1 mmol/l), but not following 100 mg (7.3 +/- 0.3), compared with placebo (7.6 +/- 0.3). Peak insulin levels were similarly delayed and reduced by the 200 mg dose only (36.5 +/- 6.1 mU/l, compared with 49.8 +/- 8.7). Suppression of nocturnal GH by 200 mg pirenzepine at night produced a small reduction in fasting plasma glucose (5.0 +/- 0.1 mmol/l, compared with 5.3 +/- 0.1, P less than 0.02) but did not improve subsequent breakfast carbohydrate tolerance. Peak plasma glucose in NIDDMs was reduced following pirenzepine (12.4 +/- 0.9 mmol/l) compared with placebo (14.3 +/- 1.0, P less than 0.01). This reduction was equally significant in obese and non-obese groups. Peak plasma insulin was also reduced by pirenzepine (22.4 +/- 3.9 mU/l) compared with placebo (42.4 +/- 5.3, P less than 0.01). Insulin suppression was quantitatively greater in obese than in non-obese patients. Improvement in carbohydrate tolerance after pirenzepine in normal subjects is dose related and largely independent of GH suppression. Cholinergic blockade can also improve meal carbohydrate tolerance with simultaneous reduction in plasma insulin concentrations in non-insulin dependent diabetics, particularly those with obesity.


Modulation of osteoclastogenesis by fatty acids.
Clinical studies have shown that total body fat mass is related to both bone density and fracture risk and that fat ingestion reduces bone turnover. These effects are at least partially mediated by endocrine mechanisms, but it is possible that lipids might act directly on bone. We assessed the effects of broad fractions of milk lipids in osteoblasts, bone marrow, and neonatal mouse calvariae. Several milk fractions and their hydrolysates inhibited osteoclastogenesis in bone marrow cultures, so we assessed the effects of free fatty acids in this model. Saturated fatty acids (0.1-10 microg/ml) inhibited osteoclastogenesis in bone marrow cultures and RAW264.7 cells. This effect was maximal for C14:0 to C18:0 fatty acids. The introduction of greater than 1 double bond abrogated this effect; omega3 and omega6 fatty acids had comparable low activity. Osteoblast proliferation was modestly increased by the antiosteoclastogenic compounds, ruling out a nonspecific toxic effect. Active fatty acids did not consistently change expression of receptor activator of nuclear factor-kappaB ligand or osteoprotegerin in osteoblastic cells nor did they affect the activity of key enzymes in the mevalonate pathway. However, receptors known to bind fatty acids were found to be expressed in osteoblastic (GPR120) and osteoclastic (GPR40, 41, 43, 120) cells. A synthetic GPR 40/120 agonist mimicked the inhibitory effects of fatty acids on osteoclastogenesis. These findings provide a novel link between lipid and bone metabolism, which might contribute to the positive relationship between adiposity and bone density as well as provide novel targets for pharmaceutical and nutriceutical development.


[Minor fatty acids of biological fluids of urogenital organs and their significance in the diagnosis of inflammatory processes].
Minor lipid components (less than 1%), not occurring in the cells of mammals, have been detected in the profile of fatty acids (FA) in urine and vaginal fluid in females, as well as in ejaculate in males, by the method of gas chromatography and mass spectrometry. Their origin is supposedly attributed to microorganisms colonizing urogenital organs under normal conditions and in infectious pathology. Minor FA in the biological fluids of the urogenital tract (UGT) in man are compared with the FA composition of pure microbial cultures. The statistical analysis of FA concentrations in 500 specimens of UGT biological fluids indicates that lipid markers of microbial origin (in UGT inflammatory diseases) have a range of quantitative changes exceeding the limits of values characteristic of healthy persons and in a number of cases correlate with the diagnosis of the disease or the results of the bacteriological study. The reliability of the interpretation of the results of the marker analysis is confirmed by the difference in the composition of UGT microorganisms under normal conditions and in pathology, correlation with the clinical picture of pathological deviations, a decrease (to zero or normal values) in the concentration of the markers of pathogens as the result of treatment with antibiotics, as well as by the adequate composition of UGT microorganisms, determined by the marker method in comparison with the cultural biochemical method.


Contact Allergy in Danish Healthcare Workers: A Retrospective Matched Case-control Study.
Contact dermatitis in healthcare workers is a world wide problem. We conducted a retrospective observational study of the patch-test results of 1402 healthcare workers and 1402 matched controls with contact dermatitis who were treated at 3 hospitals departments in Denmark between 2007 and 2014. The primary objective was to determine whether healthcare work was associated with contact allergy to thiuram mix. Unadjusted univariate analyses revealed that healthcare work was significantly associated with occupational contact dermatitis and hand dermatitis. Contact allergy to thiuram mix was more common in healthcare workers was significantly associated with having occupational contact dermatitis, hand dermatitis and older age. In conclusion, we report here a potential problem of contact allergy to thiurams in healthcare workers with contact dermatitis. Legislative authorities may in the future focus on the use of rubber accelerators in, for example, protective gloves, which are widely used by healthcare professionals.


Improvement in postoperative mortality in elective gastrectomy for gastric cancer: Analysis of predictive factors in 1066 patients from a single centre.
Gastrectomy represents the main treatment for gastric adenocarcinoma. This procedure is associated with substantial morbidity and mortality. The aim of this study was to evaluate the postoperative mortality changes across the study period and to identify predictive factors of 30-day mortality after elective gastrectomy for gastric cancer. This was a retrospective cohort study of a prospective database from a single centre. Patients treated with an elective gastrectomy from 1996 to 2014 for gastric adenocarcinoma were included. We compared postoperative mortality between four time periods: 1996-2000, 2001-2005, 2006-2010, and 2011-2014. Univariate and multivariate analyses were applied to identify predictors of 30-day postoperative mortality. We included 1066 patients (median age 65 years; 67% male). The 30-day mortality rate was 4.7%. Mortality decreased across the four time periods; from 6.5% to 1.8% (P = 0.022). In the univariate analysis, age, ASA score, albumin <3.5, multivisceral resection, splenectomy, intrathoracic esophagojejunal anastomosis, R status, and T status were significantly associated with postoperative mortality. In the multivariate analysis, ASA class 3 (OR 10.06; CI 1.97-51.3; P = 0.005) and multivisceral resection (OR 1.6; CI 1.09-2.36; P = 0.016) were associated with higher postoperative 30-day mortality; surgery between 2011 and 2014 was associated with lower postoperative 30-day mortality (OR 0.55; CI 0.33-0.15; P = 0.030). There was a decrease in postoperative 30-day mortality during this 18-year period at our institution. We have identified ASA score and multivisceral resection as predictors of 30-day mortality for elective gastrectomy for cancer.


Audiometric evaluation short and medium term in cochlear implants.
Our purpose is report the results of cochlear implant program in this Institute, since our first surgery from November 2007, until December 2012. A cross-sectional study, observational, descriptive, analyzing the information about thresholds before and after implantation, using patients files (diagnosis, onset of hearing loss, brainstem auditory evoked potential (BAEP), computed tomography (CT), magnetic resonance imaging (MRI), implanted ear, brand and model of cochlear implants (CI) and audiometric studies before and after the CI. We report the evolution of 68 patients, age ranged 1 year 8 months to 39 years 3 months old. 94% patients (n = 64) had pre-lingual hearing loss being hereditary non-syndromic hearing loss the most common etiology (29.4%). 100% patients had auditory brainstem responses showing bilateral profound hearing loss, in the 77.9% type A tympanograms were obtained (Jerger's classification), and 100% had absence of stapedial reflexes and otoacoustic emissions with low reproducibility. CT reported as normal in 85.2% of patients, the findings: 5.8% had chronic mastoiditis changes, other findings reported in 1.4% of patients were: digastric right facial nerve, facial nerve canal dehiscence, enlarged vestibular aqueduct, occupation and poor pneumatization of mastoid air cells, lateral semicircular canals agenesis, incomplete partition of the cochlea with wide vestibular and vestibular aqueduct dilatation. Most frequent MR findings of skull with cerebellopontine angle approach were vascular loops of internal auditory canals unilaterally. In 10.2%, 55.8% of patients (n = 38) were implanted in the right ear, 56 (82.3%) with a CI from Advanced Bionics, HiRes 90K model, the remaining with Cochlear, Freedom and Nucleus 5 models. Developments in CI results by audiometric tests: prior to placement was 106.2 dB averages at frequencies assessed, one month later 62.4 dB, at 6 months 44 dB, and with satisfactory threshold 32.9 dB. 55.8% of patients (n = 38) with P + HiRes Fidelity 120 strategy, the remaining with Hires S + Fidelity 120, Hires S and ACE RE. Audiology service proposed to place the CI in the worst ear by threshold in audiometric tests, the otolaryngology service proposed the best ear from anatomical point view. Implanted in the INR more Advanced Bionics CI faq frequently due to the donation by the insurance for a new generation. Hearing thresholds using CI have improved since activation.


Cryosurgery and laser surgery in the treatment of malignant and benign laryngeal processes.
Cryosurgery of the larynx is still in the stage of development. In the treatment of laryngeal papillomas cryosurgery has been quite successful. The use of cryosurgery in laryngo-hypopharyngeal carcinoma and in laryngeal stenosis is still restricted to special indications (old age, risk patients). A combination of cryosurgery and telecobalt irradiation has proven valuable in the management of tumors. After the basic physics of laser radiation are explained, the principle mechanisms of interaction between the CO2 laser on human tissue are mentioned. This is followed by the description of the different indications for laser surgery in the larynx. A chordectomy and the opening of a subglottic stenosis with the laser beam are described and documented with photographs. Finally, there is an outlook to an eventually broader spectrum of indications for laser surgery in the field of ent.


Transformation of cells overexpressing a tyrosine kinase by phospholipase D1 and D2.
Phospholipase D (PLD) activity is elevated in response to most mitogenic signals. Two mammalian PLD genes (PLD1 and PLD2) have been cloned and their gene products have been characterized. PLD1 is a downstream target of the Ras/RalA GTPase cascade implicated in mitogenic and oncogenic signaling. Consistent with a role in mitogenic signaling, elevated expression of PLD1 transforms cells overexpressing the epidermal growth factor (EGF) receptor (EGFR). However, PLD2 colocalizes with the EGFR in caveolin-enriched light membrane microdomains. We therefore investigated whether PLD2 could also contribute to the transformation of cells overexpressing a tyrosine kinase. We report here that elevated expression of PLD2 transforms rat fibroblasts overexpressing either the EGFR or c-Src. Since overexpression of a tyrosine kinase is a common genetic alteration in several human cancers, these data suggest that elevation of either PLD1 or PLD2 may contribute to the progression to a malignant phenotype in cells with elevated tyrosine kinase activity.


Migrating suture masquerading as a renal pelvic carcinoma: an unusual complication of the Kock pouch.
Following a radical cystectomy and the creation of a Kock pouch, a filling defect developed in the renal pelvis that appeared to be a transitional cell carcinoma. This appearance was produced by the retrograde migration and implantation of a fragment of suture material used in the creation of the pouch. It represents a previously undescribed complication of this procedure.


pcaH, a molecular marker for estimating the diversity of the protocatechuate-degrading bacterial community in the soil environment.
Microorganisms degrading phenolic compounds play an important role in soil carbon cycling as well as in pesticide degradation. The pcaH gene encoding a key ring-cleaving enzyme of the beta-ketoadipate pathway was selected as a functional marker. Using a degenerate primer pair, pcaH fragments were cloned from two agricultural soils. Restriction fragment length polymorphism (RFLP) screening of 150 pcaH clones yielded 68 RFLP families. Comparison of 86 deduced amino acid sequences displayed 70% identity to known PcaH sequences. Phylogenetic analysis results in two major groups mainly related to PcaH sequences from Actinobacteria and Proteobacteria phyla. This confirms that the developed primer pair targets a wide diversity of pcaH sequences, thereby constituting a suitable molecular marker to estimate the response of the pca community to agricultural practices.


Anti-inflammatory effect of a novel synthetic compound 1-((4-fluorophenyl)thio)isoquinoline in RAW264.7 macrophages and a zebrafish model.
The compound, 1-((4-fluorophenyl)thio)isoquinoline (FPTQ), is a synthetic isoquinoline derivative. To test the anti-inflammatory effect of FPTQ, we used neutrophil-specific transgenic zebrafish Tg(mpx::EGFP)i114 line and lipopolysaccharide (LPS)-stimulated RAW264.7 cells. We also used two different methods, involving tail transection and LPS stimulation in the zebrafish model. Neutrophils translocation in the zebrafish tail-transected model was inhibited by FPTQ. Neutrophil aggregation was also inhibited by FPTQ in the LPS-stimulated zebrafish model. Decreased mRNA expression of the pro-inflammatory cytokine genes, interleukin-1β (il-1β) and interleukin-6 (il-6), was found in zebrafish larvae injected with FPTQ. Additionally, production of nitric oxide was inhibited by FPTQ in RAW264.7 macrophage cells treated with LPS. Moreover, the mRNA expression of Il-1β and Il-6 suppressed by FPTQ treatment in RAW264.7 macrophage cells, and an enzyme immunoassay showed that FPTQ suppressed the secretion of IL-1β and IL-6 in RAW264.7 cells. These results demonstrate that FPTQ reduced inflammatory responses and, therefore, suggest that it may be effective as an anti-inflammatory agent.


Phenylalkylamines with potential psychotherapeutic utility. 2. Nuclear substituted 2-amino-1-phenylbutanes.
A series of 2-amino-1-(4-substituted-2,5-dimethoxyphenyl)butanes (Table V) was prepared as analogues of (R)-2-amino-1-(2,5-dimethoxy-4-methylphenyl)butane (1a). 1-(2,5-Dimethoxyphenyl)-2-(N-phthalimido)butane (7) was utilized as a synthetic intermediate common to many of the target compounds. Animal data are presented indicating that most of these analogues have low hallucinogenic potential. Selected compounds were compared with 1a in an avoidance-response acquisition model which differentiates between 1a and the human hallucinogens DOM (2a) and DOET (2b). Structure-activity relationships of these analogues are discussed.


Infection complication portends poor prognosis in acute myocardial infarction.
To determine both the incidence and impact of infectious complications of acute myocardial infarction (AMI) on length of hospital stay and mortality. This is a retrospective, case-control clinical trial involving medical records review. The study population consisted of patients admitted to the Coronary Care Unit (CCU) of the Hospital das Clínicas Heart Institute of the University of São Paulo Medical School--FMUSP--with AMI between January 1996 and December 1999. One thousand two hundred and twenty-seven patients were analyzed, and 60 (5%) met diagnostic criteria for infectious complication of AMI (infected group). The other 1167 patients served as control group. Mean age (67.5 versus 62.6), hospital length of stay (26.6 versus 12.0 days), and in-hospital mortality (45% versus 12%) were higher in the infected group. Mortality rate was higher among patients who underwent more than three invasive procedures (68% and 32%, p = 0.006). The most frequent infections were pulmonary (63%), urinary tract (37%) and positive blood cultures with no identifiable site of infection (8%). In the population studied, infectious complication rate was 5%. Prolonged hospital stay and high mortality rate suggest that infection complication has a great impact on AMI patients admitted to the coronary care unit.


The relationship between health behavior and pain scale in patients with low back pain in Thailand.
The purpose of the study was to evaluate the relationship between health behavior and pain scale in patients with low back pain. One hundred eighty patients with low back pain attending the Orthopedic Unit at Sapasithiprasong Hospital in Thailand participated in the study. The participants completed a questionnaire related to demographic data and preventive health behavior, illness health behavior and sick role health behavior. Statistical analysis was used to calculate means, standard deviations and percentages, multiple regression evaluated the relationship between demographic data and pain scale, and Pearson's correlation assessed the relationship between the three health behaviors and pain scale. Generally, the results showed most participants had a lower quality of health behavior. The most common causes of low back pain were lifting heavy loads, incorrect positioning and everyday activity (43.9, 17.8 and 10.6%, respectively). The level of participant's income was found to be statistically relevant to pain scale (p < 0.05). There was also a statistically significant association between the three health behaviors and pain scale (p = 0.0001, 0.005, 0.0001, respectively). The findings of this study illustrate the crucial role that Thai health care professionals play in changing the health behavior of patients with low back pain in order to improve the patient's quality of life.


Hepatitis E virus: A potential threat for patients with liver disease and liver transplantation.
Immunocompromised patients are at risk of acquiring acute hepatitis E virus infection (HEV), leading to chronicity. Chronic HEV infection is associated with persistent viraemia, raised transaminase activity, histological features associated with chronic hepatitis and evidence of rapid development of cirrhosis. Extrahepatic manifestations have been associated with HEV. Most frequently reported are neurological disorders with predominantly involvement of the peripheral nervous system. In patients using immunosuppressive drugs antibody production is often delayed and HEV RNA detection is superior to serology to detect infection. Therapeutic options for chronic HEV includes tapering immunosuppressive and secondly ribavirin, pegylated interferon alpha (PEG-IFN). Present recommendation is to treat chronic HEV patients for 3 months, asses serum HEV RNA and stool HEV RNA and stop therapy if both are undetectable. Studies are required to determine which other antiviral agents than ribavirin and (PEG-)IFN are of clinical utility in treating HEV in the minority of patients who do not respond to ribavirin.


Inhibition of gene expression of carnitine palmitoyltransferase I and heart fatty acid binding protein in cyclophosphamide and ifosfamide-induced acute cardiotoxic rat models.
This study investigated whether cyclophosphamide (CP) and ifosfamide (IFO) therapy alters the expression of the key genes engaged in long-chain fatty acid (LCFA) oxidation outside rat heart mitochondria, and if so, whether these alterations should be viewed as a mechanism during CP- and IFO-induced cardiotoxicity. Adult male Wistar albino rats were assigned to one of the six treatment groups: Rats in group 1 (control) and group 2 (L-carnitine) were injected intraperitoneal (i.p.) with normal saline and L-carnitine (200 mg/kg/day), respectively, for 10 successive days. Animals in group 3 (CP group) were injected i.p. with normal saline for 5 days before and 5 days after a single dose of CP (200 mg/kg, i.p.). Rats in group 4 (IFO group) received normal saline for 5 successive days followed by IFO (50 mg/kg/day, i.p.) for 5 successive days. Rats in group 5 (CP-carnitine supplemented) were given the same doses of L-carnitine as group 2 for 5 days before and 5 days after a single dose of CP as group 3. Rats in group 6 (IFO-carnitine supplemented) were given the same doses of L-carnitine as group 2 for 5 days before and 5 days concomitant with IFO as group 4. Immediately, after the last dose of the treatment protocol, blood samples were withdrawn and animals were killed for biochemical, histopathological and gene expression studies. Treatment with CP and IFO significantly decreased expression of heart fatty acid binding protein (H-FABP) and carnitine palmitoyltransferase I (CPT I) genes in cardiac tissues. Moreover, CP but not IFO significantly increased acetyl-CoA carboxylase2 mRNA expression. Conversely, IFO but not CP significantly decreased mRNA expression of malonyl-CoA decarboxylase. Both CP and IFO significantly increased serum lactate dehydrogenase, creatine kinase isoenzyme MB and malonyl-CoA content and histopathological lesions in cardiac tissues. Interestingly, carnitine supplementation completely reversed all the biochemical, histopathological and gene expression changes induced by CP and IFO to the control values, except CPT I mRNA, and protein expression remained inhibited by IFO. Data from the current study suggest, for the first time, that (1) CP and IFO therapy is associated with the inhibition of the expression of H-FABP and CPT I genes in cardiac tissues with the consequent inhibition of mitochondrial transport and oxidation of LCFA. (2) The progressive increase in cardiotoxicity enzymatic indices and the decrease in H-FABP and CPT I expression may point to the possible contribution of these genes to CP- and IFO-induced cardiotoxicity.


A novel PTPN11 mutation in LEOPARD syndrome.
PTPN11 gene mutations are common to both patients with Noonan (NS) and LEOPARD syndrome (LS). So far only two recurrent mutations have been identified in LS patients by different research groups, i.e., Tyr279Cys and Thr468Met. In this work we describe the third PTPN11 mutation that has been found in a single LS patient. The mutation (c.1517A>C) substitutes a proline for a glutamine at amino acid 506 (Gln506Pro) in the phosphatase domain (PTP) of the PTPN11 peptide SHP2. This region is a mutation hotspot. Changes at amino acids 501 to 504 cause NS. Gln506Pro is predicted, by modeling analysis, to seriously disrupt the normal contacts between the regulating N-SH2 and the active PTP domains, leading to hyperactivity of the phosphatase. This report demonstrates that rarer mutations other than Tyr279Cys and Thr468Met can be found in LS patients and the need of screening the whole gene in those negative for the commonest mutations.


Stochastic model of Zipf's law and the universality of the power-law exponent.
We propose a stochastic model of Zipf's law, namely a power-law relation between rank and size, and clarify as to why a specific value of its power-law exponent is quite universal. We focus on the successive total of a multiplicative stochastic process. By employing properties of a well-known stochastic process, we concisely show that the successive total follows a stationary power-law distribution, which is directly related to Zipf's law. The formula of the power-law exponent is also derived. Finally, we conclude that the universality of the rank-size exponent is brought about by symmetry between an increase and a decrease in the random growth rate.


No place for any prejudice.
They are vital links in the caring chain. Without them, staffing levels in many areas would probably collapse. They often work in departments with particular recruitment problems, on night duty or as bank nurses.


[Viewing of horror and violence videos by adolescence. A study of speech samples of video consumers with the Gottschalk-Gleser Speech Content analysis].
In 1990 pupils of different schools in Württemberg were interviewed about their television and video consumption. It turned out that a high percentage of mainly male pupils of Hauptschulen (upper division of elementary schools) and special schools excessively and regularly consumed films which were on the index (X-rated) or seized depicting horror and violence. Subsequent to the inquiry through questionnaires and different personality tests, speech samples of 51 test persons were recorded on tape. 5 speech samples had to be excluded from further investigation since they contained less than 70 words. The transcribed and anonymized records were examined according to the Gottschalk-Gleser content analysis of verbal behavior, and two groups of so-called seldom lookers (n = 22) and frequent lookers (n = 24) were compared to each other. The frequent lookers significantly often reported about film contents which presumably means that their imagination is more restricted and less productive than that of the other group. In addition, this group of frequent lookers had significantly higher scores concerning death anxiety and guilt anxiety. With regard to hostility affects, their scores were also significantly raised concerning outward-overt hostility, outward-covert hostility, and ambivalent hostility. Probably the group of frequent lookers comprised more test persons with relationship disorders, with borderline risks, dissocial personality features, and problems to cope with their aggressiveness. So they show on the one hand a raised affinity to watch such films, but simultaneously unconscious and conscious learning processes take place which stimulate further aggressive fantasies (and possibly also actions).


[Glomerulopathy associated with glomerular microtubular deposits: a report of a case].
A male aged 58 was admitted to our hospital because of proteinuria, hematuria and bilateral pretibial edema. Laboratory tests showed normocytic, normochromic anemia and moderately impaired renal function. Antinuclear antibodies were negative. Neither M-protein nor Bence-Jones protein were detected. Light microscopic study on the biopsied renal specimen indicated a moderate mesangial proliferation accompanying with the deposition of PAS-positive and Congo red-negative materials in the subendothelial area. C3 accumulated segmentally along the capillary walls, which was clarified by immunofluorescence microscopy. Staining for IgG, IgA, IgM and light chains were negative. Electron microscopy demonstrated the deposition of microtubules in the mesangial, subepithelial and subendothelial areas. The diameter of these microtubules ranged from 40 to 80 nm. Such type of the microtubules have been reported to exist in the glomeruli in the patients with systemic diseases such as amyloidosis, systemic lupus erythematosus, cryoglobulinemia and light chain disease. In our patient, however, any clinical or serological findings suggestive of these systemic diseases were not obtained. On the other hand previous report pointed out that microtubules deposited in the glomeruli in the patients with immunotactoid glomerulopathy or other glomerulopathies. Our patient had the clinical features consistent with these glomerulopathies. However, no depositions of immunoglobulins were observed. This case is an atypical glomerulopathy accompanying with the glomerular microtubular deposits.


[Analysis of the scales of measures, the efficacy of the treatments and the health/social impact of Alzheimer disease in the Basque Country, Spain].
The purpose of this study is to review the clinical analyses regarding efficacy of the different drugs used in the treatment of Alzheimer, across distinct scales of measures in order out the progression of the Alzheimer's Disease (AD). Along with this an economic analysis of the health/social impact of this illness in the Basque Country has been carried out. A selection and synthesis of the relevant scientific evidence was made by means of a bibliographical research in the main data bases (Medline, Embase, Cross, Iowa Drug Information System, Healthstar and The Cochrane Library). The value of the costs was carried out from a social perspective. During 1998 a health social costs analysis of this pathology was undertaken in the Basque Country, Spain, starting from a table of assumptions. ADAS and PDS are the only cognitive and functional scales developed and validated specifically for the AD. The drugs specifically prescribed for the AD (tacrine, donepezil, and revastigmine) contribute a slight benefit, whose clinical meaning is unknown. During 1998 the global cost of treating the AD in the Basque Country has been at least 55,858 million pesetas (US$ 360.5 millions dollars). The social cost (nursing homes, day centres, home help, help from family and friends) comes to represent 98.4% of this global cost. At the moment, there is not any effective drug in the treatment of the AD. The informal cost (help from family and friends) comes to represent 88.4% of the total social costs.


The role of Müllerian inhibiting substance in the evaluation of phenotypic female patients with mild degrees of virilization.
Müllerian inhibiting substance (MIS) is a sexually dimorphic gonadal hormone with proven efficacy in the evaluation of boys with cryptorchidism and children with intersex conditions. We examined the role of MIS determination in the evaluation of 65 phenotypic females with mild virilization. Among the 28 subjects with MIS values elevated above the normal female range, all had abnormal gonadal tissue: ovotestes in 11, testes in 7, dysgenetic gonads in 7, and MIS-secreting ovarian tumors in 3. Among the 37 children with serum MIS in the normal female range, 19 had detectable MIS and 18 had unmeasurable MIS. In the former group with measurable but normal female MIS values, 16 subjects had ovaries, 1 had an ovotestis, and 1 had dysgenetic gonads containing testicular elements. Of 18 children with undetectable MIS values, 16 had ovaries and 2 had ovarian dysgenesis. In this study, elevation of serum MIS above the normal female range was consistently associated with the presence of testicular tissue or MIS- secreting tumors, mandating additional evaluation and surgical exploration. A value within the normal female range in a virilized patient did not exclude dysgenetic testicular tissue or ovotestis, whereas undetectable values were consistent with the absence of testicular tissue.


Resection of World Health Organization Grade II gliomas involving Broca's area: methodological and functional considerations.
Advances in functional mapping have enabled us to extend the indications of surgery for low-grade gliomas (LGGs) within eloquent regions. However, to our knowledge, no study has been specifically dedicated to the resection of LGGs within Broca's area. We report the first surgical series of LGGs involving this area by focusing on methodological and functional considerations. Seven patients harboring an LGG in Broca's area (revealed by partial seizures) had a language functional magnetic resonance imaging scan and then underwent operation while awake using intrasurgical electrical mapping. The neurological examination was normal in all patients despite mild language disturbances shown using the Boston Diagnosis Aphasia Examination. Both pre- and intraoperative cortical mapping found language reorganization with recruitment of the ventral and dorsal premotor cortices, orbitofrontal cortex, and insula, whereas no or few language sites were detected within Broca's area. Subcortically, electrostimulation allowed the identification and preservation of four structures still functional, including the arcuate fasciculus, fronto-occipital fasciculus, fibers from the ventral premotor cortex, and head of the caudate. Postoperatively, after transient language worsening, all patients recovered and returned to a normal socioprofessional life. The resection was total in three cases, subtotal in three, and partial in one patient (operated twice). Our results indicate that, in patients with no aphasia despite LGGs within Broca's area, thanks to brain plasticity, the tumor can be removed while involving this "unresectable" structure without inducing sequelae and even improving the quality of life when intractable epilepsy is relieved on the condition that subcortical language connectivity is preserved.


Engineering a recombinant Deinococcus radiodurans for organopollutant degradation in radioactive mixed waste environments.
Thousands of waste sites around the world contain mixtures of toxic chlorinated solvents, hydrocarbon solvents, and radionuclides. Because of the inherent danger and expense of cleaning up such wastes by physicochemical methods, other methods are being pursued for cleanup of those sites. One alternative is to engineer radiation-resistant microbes that degrade or transform such wastes to less hazardous mixtures. We describe the construction and characterization of recombinant Deinococcus radiodurans, the most radiation-resistant organism known, expressing toluene dioxygenase (TDO). Cloning of the tod genes (which encode the multicomponent TDO) into the chromosome of this bacterium imparted to the strain the ability to oxidize toluene, chlorobenzene, 3,4-dichloro-1-butene, and indole. The recombinant strain was capable of growth and functional synthesis of TDO in the highly irradiating environment (60 Gy/h) of a 137Cs irradiator, where 5x10(8)cells/ml degraded 125 nmol/ml of chlorobenzene in 150 min. D. radiodurans strains were also tolerant to the solvent effects of toluene and trichloroethylene at levels exceeding those of many radioactive waste sites. These data support the prospective use of engineered D. radiodurans for bioremediation of mixed wastes containing both radionuclides and organic solvents.


Effects of ambient illumination, luminance contrast, and stimulus type on subjective preference of VDT target and background color combinations.
This study investigated the effects of the target and background color combination on subjective preference, i.e., aesthetic appearance, legibility and visual comfort, of stimuli presented on a visual display terminal under various ambient illuminations, luminance contrasts, and stimulus types. Analysis showed the main effect of color combination was significant for the three dependent variables. Black-on-white and blue-on-white were the most preferred combinations, while turquoise-on-green and turquoise-on-red were the least preferred. A sign was rated better than text. The significant interaction between color combination and luminance contrast indicated that more preferred color combinations were rated better under higher luminance contrast, whereas less preferred color combinations had better ratings under lower luminance contrast.


Downhill running: a model of exercise hyperemia in the rat spinotrapezius muscle.
To utilize the rat spinotrapezius muscle as a model to investigate the microcirculatory consequences of exercise training, it is necessary to design an exercise protocol that recruits this muscle. There is evidence that the spinotrapezius is derecruited during standard treadmill exercise protocols performed on the uphill treadmill (i.e., 6 degrees incline). This investigation tested the hypothesis that downhill running would effectively recruit the spinotrapezius muscle as assessed by the presence of an exercise hyperemia response. We used radioactive 15-microm microspheres to determine blood flows in the spinotrapezius and selected hindlimb muscles of female Sprague-Dawley rats at rest and during downhill (i.e., -14 degrees incline; 331 +/- 5 g body wt, n = 7) and level (i.e., 0 degrees incline; 320 +/- 11 g body wt, n = 5) running at 30 m/min. Both level and downhill exercise increased blood flow to all hindlimb muscles (P < 0.01). However, in marked contrast to the absence of a hyperemic response to level running, blood flow to the spinotrapezius muscle increased from 26 +/- 6 ml.min(-1).100 g(-1) at rest to 69 +/- 8 ml.min(-1).100 g(-1) during downhill running (P < 0.01). These findings indicate that downhill running represents an exercise paradigm that recruits the spinotrapezius muscle and thereby constitutes a tenable physiological model for investigating the adaptations induced by exercise training (i.e., the mechanisms of altered microcirculatory control by transmission light microscopy).


Proarrhythmic and antiarrhythmic effects of intravenous prostacyclin in man.
Prostacyclin (PGI2) has been shown to reduce the occurrence of experimental ventricular arrhythmias. To assess potential beneficial effects in man, the electrophysiological action of PGI2 was studied in 16 non medicated patients. The protocol used in incremental pacing and programmed stimulation in the right atrium and ventricle. This protocol and measurement of effective refractory periods (ERP) were performed before and during the injection of 2.5, 5 and 10 ng kg-1 min-1 of PGI2. The atrial functional refractory period decreased significantly (P less than 0.05); PGI2 had no influence on the occurrence of inducible non-sustained (NS) atrial tachycardias and was responsible for the occurrence of 2 non-sustained atrial tachycardias in 8 patients with inducible atrial echo beats under basal conditions. Thirteen patients did not have inducible ventricular tachycardia (VT) under basal conditions. Non-sustained VT was induced after PGI2 in 4 of them but in only 1 of them after the administration of propranolol. Three patients had inducible VT under basal conditions (1 non-sustained, 2 sustained VT). PGI2 did not prevent the occurrence of VT (1 non-sustained, 1 sustained VT), except in 1 patient with ischaemic-related VT, who had non-sustained VT after PGI2. In conclusion, PGI2 does not seem to have a cardiac antiarrhythmic effect and may increase the atrial and ventricular repetitive response. This effect could be related to an increase of adrenergic tone.


Clinical characteristics and cardiovascular magnetic resonance findings in stress (takotsubo) cardiomyopathy.
Stress cardiomyopathy (SC) is a transient form of acute heart failure triggered by stressful events and associated with a distinctive left ventricular (LV) contraction pattern. Various aspects of its clinical profile have been described in small single-center populations, but larger, multicenter data sets have been lacking so far. Furthermore, it remains difficult to quickly establish diagnosis on admission. To comprehensively define the clinical spectrum and evolution of SC in a large population, including tissue characterization data from cardiovascular magnetic resonance (CMR) imaging; and to establish a set of CMR criteria suitable for diagnostic decision making in patients acutely presenting with suspected SC. Prospective study conducted at 7 tertiary care centers in Europe and North America between January 2005 and October 2010 among 256 patients with SC assessed at the time of presentation as well as 1 to 6 months after the acute event. Complete recovery of LV dysfunction. Eighty-one percent of patients (n = 207) were postmenopausal women, 8% (n = 20) were younger women (aged ≤50 years), and 11% (n = 29) were men. A stressful trigger could be identified in 182 patients (71%). Cardiovascular magnetic resonance imaging data (available for 239 patients [93%]) revealed 4 distinct patterns of regional ventricular ballooning: apical (n = 197 [82%]), biventricular (n = 81 [34%]), midventricular (n = 40 [17%]), and basal (n = 2 [1%]). Left ventricular ejection fraction was reduced (48% [SD, 11%]; 95% confidence interval [CI], 47%-50%) in all patients. Stress cardiomyopathy was accurately identified by CMR using specific criteria: a typical pattern of LV dysfunction, myocardial edema, absence of significant necrosis/fibrosis, and markers for myocardial inflammation. Follow-up CMR imaging showed complete normalization of LV ejection fraction (66% [SD, 7%]; 95% CI, 64%-68%) and inflammatory markers in the absence of significant fibrosis in all patients. The clinical profile of SC is considerably broader than reported previously. Cardiovascular magnetic resonance imaging at the time of initial clinical presentation may provide relevant functional and tissue information that might aid in the establishment of the diagnosis of SC.


Inhibition of a peripheral sympathetic-cholinergic system by presynaptic alpha 2-adrenoceptors.
Intravenous administration of catecholamines produced dose-related inhibition of electrodermal responses (EDRs) evoked by electrical stimulation of the post-ganglionic sciatic nerve in anaesthetized rats, with relative potencies being (-)-adrenaline greater than (+/-)-adrenaline greater than (-)-noradrenaline = (+)-adrenaline. The suppression of EDRs by (+/-)-adrenaline and (-)-noradrenaline was blocked by pretreatment with yohimbine (0.75 mg/kg i.v.) but not by prazosin (0.3 mg/kg i.v.). The selective alpha 2-adrenoceptor agonist B-HT 920 also inhibited neurally evoked skin potential responses. This effect of B-HT 920 was antagonized by the selective alpha 2-adrenoceptor antagonist idazoxan (0.1 mg/kg i.v.) but was insensitive to prazosin. Idazoxan was more potent than yohimbine in blocking (+/-)-adrenaline-induced suppression of EDRs. Methacholine administered into the femoral artery evoked EDRs by an atropine-sensitive mechanism. Methacholine-induced EDRs were not suppressed by intravenous administration of (-)-adrenaline (1 microgram/kg or 3 micrograms/kg) whereas EDRs evoked by the sciatic nerve stimulation on the other hindpaw were inhibited. Increase in the endogenous catecholamines by asphyxia strongly inhibited EDRs by a mechanism which was also sensitive to yohimbine but not to prazosin. These results suggest that peripheral presynaptic alpha 2-adrenergic mechanisms are involved in inhibition of transmitter release in this sympathetic-cholinergic system.


Quality assurance of immunodiagnostic tests in Australasia.
A Royal College of Pathologists of Australasia (RCPA) sponsored Quality Assurance Program (QAP) in Clinical Immunology, involving 128 laboratories over a 1 yr period, revealed the following: successful participation in the program by 16 overseas laboratories (distant from Australia); only 30% of laboratories succeeded in returning their results by the scheduled date on every occasion; quantitation of urinary total protein and Bence Jones protein was poor and varied over a log scale; immunofixation was more successful in characterizing urinary paraprotein than immunoelectrophoresis; densitometry of protein electrophoresis appeared the method of choice in quantitating serum paraproteins accurately; nephelometric techniques gave better concordance between laboratories than turbidometric, radial immunodiffusion or agglutination techniques; poor concordance between laboratories in detecting weakly positive antinuclear antibodies (ANA); some laboratories had difficulties in identifying ANA patterns (only 60% of laboratories correctly identified the anticentromere pattern); few laboratories could correctly identify antibodies to extractable nuclear antigens (ENA); flow cytometry gave a smaller dispersion of lymphocyte subpopulation percentages than microscopy. A method was established to rank laboratory performance of selected tests over the 1 yr period. Such a comparative ranking scheme may alert laboratories in identifying specific or generalized deficiencies in performance.


Enthalpic and entropic contributions to actin stability: calorimetry, circular dichroism, and fluorescence study and effects of calcium.
The delta H associated with the thermal unfolding of G-actin has been determined by differential scanning calorimetry (DSC) to be 142 +/- 5 kcal/mol, with the Tm (melting temperature) at 57.2 +/- 0.5 degrees C, at pH 8.0 (heating rate 0.5 K/min). The transition is broad and cannot be treated as a single transition that mimics a two-state process, suggesting the existence of domains. Deconvolution is done to fit it into two quasi-independent two-state transitions. For F-actin, the transition is more cooperative, with a cooperative ratio (the ratio of van't Hoff enthalpy and calorimetric enthalpy) of 1.4, indicating intermonomer interaction. The delta H of the thermal unfolding of F-actin is 162 +/- 10 kcal/mol with a Tm at 67.0 +/- 0.5 degrees C. A state of G-actin similar to that of the heat-denatured form, designated D-actin, is obtained by removing tightly bound Ca2+ with EGTA. The DSC-detectable cooperative transition is completely lost when the free calcium concentration of the medium is 1 x 10(-11) M or lower, using a Ca2+/EGTA buffer system. However, circular dichroism (CD) shows that the helix content of actin, 32% in the G-form, is only partially reduced to 19% in this apo form. The CD spectrum and the helix content of the calcium-depleted actin are almost identical with those of the heat-denatured D form. This loss of 40% of the native helical content is irreversible in both cases. The remaining 60% of the native helical content cannot be further eliminated by heating to 95 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)


Exploring the catalytic core of complex I by Yarrowia lipolytica yeast genetics.
We have developed Yarrowia lipolytica as a model system to study mitochondrial complex I that combines the application of fast and convenient yeast genetics with efficient structural and functional analysis of its very stable complex I isolated by his-tag affinity purification with high yield. Guided by a structural model based on homologies between complex I and [NiFe] hydrogenases mutational analysis revealed that the 49 kDa subunit plays a central functional role in complex I. We propose that critical parts of the catalytic core of complex I have evolved from the hydrogen reactive site of [NiFe] hydrogenases and that iron-sulfur cluster N2 resides at the interface between the 49 kDa and PSST subunits. These findings are in full agreement with the "semiquinone switch" mechanism according to which coupling of electron and proton transfer in complex I is achieved by a single integrated pump comprising cluster N2, the binding site for substrate ubiquinone, and a tightly bound quinone or quinoid group.


Relationships among diabetes, microalbuminuria, and ACE inhibition.
Increasing worldwide rates of diabetes mellitus, combined with the significant micro- and macrovascular complications that accompany the disease, point to a heightened need to develop simple, rational strategies to protect against end-organ damage, particularly diabetic nephropathy. Although simple strategies do exist, i.e., early diagnosis of microalbuminuria (MAU) followed by nephroprotective therapy with angiotensin-converting enzyme (ACE) inhibitors, the use of these techniques should be increased. This is especially true for primary care physicians, who will continue to handle the majority of diabetic patients. To combat the underuse of this dual approach, this article reviews a stepwise approach to identifying early MAU so that therapeutic measures can be undertaken before the disorder progresses to diabetic nephropathy and, consequently, to end-stage renal disease in the majority of patients. Once a diagnosis of MAU is made, a variety of trials have demonstrated that ACE inhibitor therapy should be considered the standard therapy to retard worsening albuminuria and subsequent renal disease. ACE inhibitors can retard the progression of microalbuminuria and can lower the percentage of patients who progress to end-stage renal disease and death. Significant data indicate that ACE inhibitors should be used in MAU diabetics regardless of the level of blood pressure. In particular, the ACE inhibitor fosinopril may offer advantages because of its dual route of elimination, thus simplifying dosing in renally impaired patients. The newly developed angiotensin II receptor antagonists should be considered in patients intolerant to ACE inhibitors because of the similar effect on the interruption of the renin-angiotensin system.


Uptake and distribution of intravenously or intravesically administered photosensitizers in the rat.
Photodynamic therapy using i.v. injected porphyrin photosensitizers have been used to treat selected cases of superficial bladder cancer. Since cutaneous photosensitivity, lasting 6-8 weeks, is a well known undesirable side effect of this therapy, we instilled the photosensitizers intravesically in rats and compared the uptake of photosensitizers in different tissues by this route of administration with the uptake after intravenous injection. The intravesical mode of delivery enhanced photosensitizer uptake in the bladder wall, while giving low concentrations in extravesical organs. Intravesical instillation of the photosensitizers may therefore increase their efficacy and reduce phototoxicity as compared with intravenous injection. Comparing the results obtained by two assays, one based on porphyrin fluorescence and the other based on the application of radioactively labelled photosensitizers, it was concluded that the i.v. administration route may result in tissue uptake of significant amounts of aggregated non-fluorescent, supposedly inactive drug, while the intravesical administration led to less uptake of aggregates relative to active drug monomers.


No evidence of infection with porcine endogenous retrovirus in recipients of porcine islet-cell xenografts.
The study of whether porcine xenografts can lead to porcine endogenous retrovirus (PERV) infection of recipients is critical for evaluating the safety of pig-to-man xenotransplantation. PERV is carried in the pig germline, and all recipients of porcine tissues or organs will be exposed to the virus. We studied 10 diabetic patients who had received porcine fetal islets between 1990 and 1993, looking for evidence of PERV infection by using PCR serology, PCR, and reverse transcriptase assays. Prolonged xenograft survival (up to a year) was confirmed in five patients by porcine C-peptide excretion and detection of pig mitochondrial DNA (mtDNA) in serum. Despite the evidence for extended exposure to pig cells and despite concomitant immunosuppressive therapy, we were unable to detect markers of PERV infection in any patient. Screening for two PERV sequences in peripheral blood lymphocytes collected 4-7 years after the xenotransplantation was negative. Markers of PERV expression, including viral RNA and reverse transcriptase, were undetectable in sera from both early (day 3 to day 180) and late (4-7 years) time points. Western blot analysis for antibodies was consistently negative. These results suggested the absence of PERV infection in these patients. Also this study establishes a minimum standard for post-transplant surveillance of patients given porcine xenografts.


Lead levels determined in Swedish permanent teeth by particle-induced X-ray emission.
The determination of lead in permanent teeth is a useful measure of past exposure in early childhood since these teeth are mineralized in early childhood. Particle-induced X-ray emission (PIXE) analysis has been shown to be a method with good applicability for the contamination-free analysis of elements heavier than calcium in dental hard tissues. The method is rapid and nondestructive. The purpose of this study, which used the PIXE technique, was to survey the average level of lead in the coronal dentin of permanent bicuspid teeth collected in three places representing Swedish urban and rural areas. In addition teeth from the New York City area were analyzed. The material comprised 165 teeth from Sweden and, for comparison, 14 teeth from New York City. The median value of lead in the Swedish teeth was 2.9 micrograms/g, a value indicating an insignificant influence from the environment in comparison to the New York teeth, for which the median value was 9.2 micrograms/g. There was however a statistically significant difference in the lead concentration of teeth from large and small Swedish cities; this finding may reflect different automobile traffic intensity.


Surgical and hardware complications of deep brain stimulation. A seven-year experience and review of the literature.
Deep brain stimulation (DBS) has been established as a safe and efficient method for the treatment of various movement disorders. As the emerging applications continue to expand and more centers become eligible for the procedure, complication rates and complication avoidance become increasingly important. Our aim was to report the DBS-related complication in our department over the last 7 years, compare our rates with those reported in the literature, and highlight those practices that will aid complications avoidance. Since 2003, 106 patients underwent DBS for various pathologies in our department. There were 38 (36%) females and 68 (64%) males with a mean age of 57 years. Preoperative diagnoses included Parkinson's disease (n = 88), dystonia (n = 12), tremor (n = 3), epilepsy (n = 1), obsessive-compulsive disorder (n = 1), and central pain syndrome (n = 1). Surgical and hardware-related complications, their treatment, and outcome were recorded and compared with those reported in the literature. There were 12 procedure-related complications (11.3% of patients, 5.7% of the procedures). These included death (n = 1), aborted procedure (n = 1), postoperative respiratory distress (n = 3), intracranial hemorrhage (n = 2), epilepsy (n = 1), postoperative confusion or agitation (n = 3), and malignant neuroleptic syndrome (n = 1). Hardware-related complications presented in 4.3% of the procedures and included infection (five patients, 4.7%), electrode breakage (0.94%), lead migration or misplacement (0.94%), and stricture formation (two patients, 1.9%). Complication rates after DBS surgery remain low, proving that DBS is not only effective but also safe. Certain strategies do exist in order to minimize complications.


[Analysis of handwriting by the dynamic programming matching].
We examined the temporal invariance hypothesis in handwriting, using dynamic programming (DP) matching algorithm. The DP matching algorithm was originally developed for the speech recognition to eliminate fluctuations of time axis caused by different speech rates. This algorithm can determine the optimal point-to-point correspondences (warping function) between the standard velocity pattern and different patterns. We found that this algorithm worked well for various velocity patterns, even when their total writing time was different from the standard pattern. Only 30 to 50 percent of handwriting movements showed temporal invariance. The writing movements which didn't show temporal invariance consisted of several sub-motor units. The borders between the units were not limited near the end of each letter. The results suggest that handwriting of a word does not show the temporal invariance and that it is executed by some discrete sub-motor units.


Distinct sites of renal fibrosis in Crim1 mutant mice arise from multiple cellular origins.
Crim1 is a transmembrane protein that regulates the bioavailability of growth factors such as VEGFA. Crim1(KST264)(/)(KST264) hypomorphic mice develop renal disease characterized by glomerular cysts and loss of endothelial integrity, progressing to peritubular and pericystic fibrosis. Peritubular capillary endothelial cells display morphological changes as well as detachment from the basement membrane. In this study, gene expression profiling of CD31(+) endothelial cells isolated from Crim1(KST264)(/)(KST264) kidneys showed up-regulation of transcripts associated with fibrosis (Col3a1, Loxl1), endothelial dysfunction (Abp1, Dcn, Lcn2), biomarkers of renal damage (Lcn2, Havcr1/Kim1) as well as evidence for a TGFβ1/TNF-associated inflammatory process. To determine whether the aberrant endothelium may in part contribute to the fibrogenic process, Tie2Cre-DsRed lineage tracing was undertaken in Crim1(KST264/KST264) mice. Approximately 31% of de novo αSMA(+) myofibroblasts detected within the tubulointerstitium were Tie2(+) DsRed(+) . However, 5.3% were F4/80(+) DsRed(+) , indicating a small population of myofibroblasts of monocytic rather than endothelial origin. In contrast, only 12% of myofibroblasts located around glomerular cysts were Tie2(+) DsRed(+) , with 7.7% being monocyte-derived (F4/80(+) DsRed(+) ). Collectively, this model supports the involvement of endothelial cells/monocytes in fibrosis within the tubulointerstitium, but also the heterogeneity of the fibrotic process even within distinct regions of the same kidney.


Regioselective di- and tetra-functionalisation of γ-cyclodextrin using capping methodology.
Alkylation of γ-cyclodextrin with 3 equiv. of 1,3-bis[bis(4-tert-butylphenyl)chloromethyl]benzene, followed by permethylation afforded selectively a singly capped (A,B), as well as two doubly capped (A,B:D,E and A,B:E,F) methylated γ-CDs. Deprotection with HBF4 gave quantitatively the corresponding diols and tetrols, which constitute valuable starting compounds for further functionalisation.


[Aging changes in conjunctiva visualized by fluorescein angiography and histopathology].
In order to investigate the aging changes of the conjunctiva, we examined fluorescein-positive areas by fluorescein angiography. The deposition of melanin in basal cells, the degranulation of mast cells, and the lumens of capillaries were examined by light microscopy in 35 cataractous eyes without diabetes or hypertension. The fluorescein leakage increased with aging. The number of basal cells with deposition of melanin granules and mast cells with degranulation correlated significantly with aging. On the other hand, the narrowing of capillary lumens in conjunctival stroma was not observed with aging. Based on the above results, we conclude that the weakness of the conjunctival vessels without the narrowing of capillary lumens might be caused by functional disorder of the endothelium and pericytes. The increase of deposition of melanin granules in basal cells may be caused by the breakdown of the metabolic pathway of melanin. Long-time mechanical stimuli and metabolic disorders of active substances may increase the degranulation of mast cells.


Mu opioid receptor gene expression in immune cells.
We have previously demonstrated that administering morphine sulfate to rhesus monkeys alters the cell-mediated as well as humoral immune responses of these primates. Furthermore, morphine treatment greatly reduces the chemotactic and phagocytotic activities of primate polymorphonuclear (PMN) cells. The present study describes the identification and isolation of mRNA encoding the mu opioid receptor gene sequence from human and monkey immune cells. Through the use of primer sequences designed from the human brain mu opioid receptor cDNA sequence, specific opioid receptor segments in mRNA transcripts were amplified, cloned, and sequenced. The mu opioid receptor gene was therefore found expressed in the following cell types: CEM x174 (a hybrid of human T and B cells), Raji (human B cells), human CD4+ cells, human monocytes/macrophages, human PMN, monkey peripheral blood mononuclear cells (PBMC), and monkey PMN. These studies present the first evidence to demonstrate that cells of human and monkey immune systems constitutively express mu opioid receptor mRNA.


Intraparenchymal NGF infusions rescue degenerating cholinergic neurons.
Nerve growth factor (NGF) exerts both trophic (cell survival) and tropic (axonal growth-promoting) effects on several neuronal populations. In particular, its robust ability to prevent lesion-induced and spontaneous age-related basal forebrain cholinergic neuronal degeneration, and to promote mnemonic recovery, has suggested its potential use as a therapeutic agent in Alzheimer's disease. When infused intracerebroventricularly, however, NGF is associated with several adverse effects that make this delivery route impractical. The present study examined whether intraparenchymal infusions of NGF adjacent to cholinergic neuronal soma are an effective and well-tolerated means of providing NGF to degenerating cholinergic neurons. Cholinergic neuronal rescue together with axonal sprouting responses and local tissue damage in the brain were assessed in adult rats that underwent complete unilateral fornix transections, followed by intraparenchymal infusions of recombinant human NGF for a 2-week period. Intraparenchymal NGF infusions prevented the degeneration of 94.7+/-6.6% of basal forebrain cholinergic neurons compared to 21.7+/-2.6% in vehicle-infused animals (p < 0.0001). Cholinergic axons sprouted toward the intraparenchymal NGF source in an apparent gradient-dependent manner. Glial responses to intraparenchymal infusions were minimal, and no apparent toxic effects of the infusions were observed. Thus, when infused intraparenchymally, NGF rescues basal forebrain cholinergic neurons, alters the topography of axonal sprouting responses, and does not induce adverse affects over a 2-week infusion period. Intraparenchymal NGF delivery merits further study at longer term time points as a means of treating the cholinergic component of neuronal loss in Alzheimer's disease.


Colchicine-induced acute myopathy in a patient with concomitant use of simvastatin.
Colchicine and 3-hydroxy-3-methy-glutaryl coenzyme A (HMG-CoA) reductase inhibitors are well known to cause myopathy. Myotoxicity is dose-dependent in both drugs; therefore, the onset of symptoms usually takes months or years. We report the case of a patient with chronic renal failure who had been taking simvastatin for 2 years and developed acute weakness 2 weeks after the start of treatment with colchicines for recurrent gout. The electromyography and elevated muscle enzymes indicated that his symptoms were caused by myopathy. When this patient stopped taking both drugs, his weakness resolved rapidly. Acute myopathy induced by combination therapy with colchicines and simvastatin is rare. In patients with chronic renal failure, co-administration of colchicine with simvastatin may accelerate the onset of myopathy because CYP3A4 (part of cytochrome P450) is crucial in the breakdown of both drugs. When adding colchicine to a medication regimen that includes a HMG-CoA reductase inhibitor for patients with renal insufficiency, drugs that are metabolized outside the CYP3A4 system (e.g., fluvastatin and pravastatin) should be selected instead.


Biophysics of radiofrequency ablation.
Radiofrequency (RF) ablation is a treatment modality that kills unwanted tissue by heat. Starting with cardiac arrhythmia treatment in the 1980s, RF ablation has found clinical application in a number of diseases, and is now the treatment of choice for certain types of cardiac arrhythmia and certain cancers. During RF ablation, an electrode is inserted into or steered intravascularly to the target tissue region under medical imaging guidance. Then, a tissue volume surrounding the electrode is destroyed by heating via RF electric current. This paper reviews the biophysics of tissue heating during RF ablation. Effects of electrical tissue conductivity and its change with temperature are discussed. Procedures and devices specific for cancer treatment and for arrhythmia treatment are presented with a brief discussion of additional clinical applications.


Motion artifact correction of multi-photon imaging of awake mice models using speed embedded HMM.
Multi-photon fluorescence microscopy (MFM) captures high-resolution anatomical and functional fluorescence image sequences and can be used for the intact brain imaging of small animals. Recently, it has been extended from imaging anesthetized and head-stabilized animals to awake and head-restrained ones for in vivo neurological study. In these applications, motion correction is an important pre-processing step since brain pulsation and tiny body movement can cause motion artifacts and prevent stable serial image acquisition at such a high spatial resolution. This paper proposes a speed embedded hidden Markov model (SEHMM) for motion correction in MFM imaging of awake head-restrained mice. The algorithm extends the traditional HMM method by embedding a motion prediction model to better estimate the state transition probability. SEHMM is a line-by-line motion correction algorithm, which is implemented within the in-focal-plane 2-D videos and can operate directly on the motion-distorted imaging data without external signal measurements such as the movement, heartbeat, respiration, or muscular tension. In experiments, we demonstrat that SEHMM is more accurate than traditional HMM using both simulated and real MFM image sequences.


[Zidovudine-associated mitochondriopathy: three possible observations in Abidjan, Côte d'Ivoire].
In Africa, prevention of mother-to-child transmission of HIV (PMTCT) with antiretrovirals is becoming a key component of the response to the pandemic. Toxicity issues remain however a concern and require careful monitoring. We report here three observations of mild neurological deterioration among children for whom a diagnosis of mitochondrial dysfunction was considered possible. These children were identified within a PMTCT research program (ANRS 049) conducted in Abidjan, Côte d'Ivoire, and evaluating a short regimen of maternal zidovudine monotherapy for PMTCT of HIV type 1. Maternal HIV-1 infection was diagnosed during pregnancy before enrolment in the randomised trial (two cases) or in the subsequent open cohort (one case). These three women had been allocated to the ZDV group and had no particular medical history. Pregnancy check-up was negative except the diagnosis of HIV-1 infection. The three children were diagnosed as uninfected by HIV-1. Symptoms developed by the age of six months (two cases) and 13 months (one case): growth failure, anthropometric abnormalities, impaired psycho-motor development, generalised and repeated seizures. The evolution of these three HIV-uninfected children was favourable after 12 to 18 months. The transient nature of these abnormalities is compatible with mild complications of mitochondrial dysfunction. We conclude however that the anticipated benefits of PMTCT with antiretrovirals in Africa greatly outweigh the potential risks and should not lead to reconsider their public health interest


On the formation of oxygenated radicals by fredericamycin A and implications to its anticancer activity: an ESR investigation.
It has been recently suggested that the exceptionally high antitumor and antibacterial activity of natural fredericamycin A (FMA) is related to its ability to spontaneously generate the superoxide anion (O2-) and hydroxyl (.OH) radicals in aerobic solutions [Hilton, B. D., Misra, R., & Zweier, J. L. (1986) Biochemistry 25, 5533]. With a view to understand the mechanistic details, attempts were made to reproduce earlier electron spin resonance (ESR) evidence for the oxygenated free radical formation in well-aerated solutions of natural FMA in dimethyl sulfoxide and dilute H2O2. Little or no evidence was obtained for the formation of the O2- and methoxy (.OCH3) radicals, while the detected formation of the .OH and methyl (.CH3) radicals was attributable largely to mechanisms not involving FMA. These results thus reopen the question regarding the mechanism of its exceptionally high tumoricidal-bacteriocidal activity.


Simultaneous determination of metformin and gliclazide in human plasma by liquid chromatography-tandem mass spectrometry: application to a bioequivalence study of two formulations in healthy volunteers.
A rapid and sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and validated to simultaneously determine gliclazide and metformin in human plasma using huperzine A as the internal standard (IS). After acetonitrile-induced protein precipitation of the plasma samples, gliclazide, metformin and the IS were subjected to LC/MS/MS analysis using electro-spray ionization (ESI). Chromatographic separation was performed on a Hypersil BDS C18 column (50 mm x 2.1 mm, i.d., 3 microm). The method had a chromatographic running time of 2.0 min and linear calibration curves over the concentration ranges of 10-10,000 ng ml(-1) for gliclazide and 7.8-4678.9 ng ml(-1) for metformin. The recoveries of the method were found to be 71-104%. The lower limits of quantification (LOQ) of the method were 10.0 and 7.8 ng ml(-1) for gliclazide and metformin, respectively. The intra- and interday precision was less than 15% for all quality control samples at concentrations of 100, 500, and 2000 ng ml(-1). The validated LC/MS/MS method has been used to study bioequivalence in healthy volunteers. These results indicate that the method was efficient with a very short running time (2.0 min) for metformin and gliclazide compared to the methods reported in the literature. The presented method had acceptable accuracy, precision and sensitivity and was used in clinical bioequivalence study.


Oxygen saturation in premature infants at risk for threshold retinopathy of prematurity.
We aimed to determine if oxygen saturation, desaturations, and saturation variability play a role in progression of retinopathy of prematurity (ROP) and need for laser treatment. This was a retrospective case-control study. We performed chart review of premature infants in a university hospital neonatal intensive care unit consecutively examined for ROP between May 2000 and December 2001. We compared birthweight, postmenstrual age, and oxygen saturation for 3 weeks before laser treatment for threshold ROP in group 1 (n=19) (average weight at treatment 2508 grams) and group 2 (n=18) before they reached 2500 grams. Outcome measures were retinopathy progression and need for treatment. Adjusting for birthweight and postmenstrual age, known predictors of ROP progression, we found that babies requiring laser treatment (group 1) had lower average daily oxygen saturation levels in the study period, significantly on 5/20 days (25%). These babies had saturations below 95% on 18/20 days (90%). Babies not requiring laser (group 2) had saturations below 95% on 2/20 days (10%). The last day on which pretreatment saturations differed significantly was 2 days before laser. Group 1 had more desaturations below 80% (6.0±3.2) than group 2 (3.2±1.2), p=0.0002 (independent samples t tests). Saturations varied more for individual group 1 infants. Decreased oxygen saturation as early as 3 weeks and as late as 2 days before laser, increased number of desaturations, and saturation variability were seen in babies eventually requiring laser treatment for ROP.