Voiding urosonography with second-generation ultrasound contrast versus micturating cystourethrography in the diagnosis of vesicoureteric reflux.

Vesicoureteric reflux has been associated with paediatric urinary tract infection. Fluoroscopic micturating cystourethrography (MCU) has been the gold standard of diagnostic test for decades; however, it has been criticized owing to its lower detection rate and radiation dose to children. Therefore, new radiation-free reflux imaging modalities have been developed, in which ultrasound-based contrast-enhanced voiding urosonography (ceVUS) is a good example. However, ultrasonography has been considered as an operator-dependent examination. Therefore, our study aimed to examine the inter-observer agreement of this sonographic technique, which has not been evaluated before. Moreover, the second-generation ultrasound contrast SonoVue has been recently marketed, and the data on its efficacy on intravesical use in ceVUS is relatively scarce. Thus, we also aimed to investigate the diagnostic performance and safety profile of SonoVue-enhanced VUS in the diagnosis of vesicoureteric reflux. Our prospective comparative study compared the diagnostic performance of ceVUS with MCU in young children presenting with first episode of urinary tract infection. We performed sequential ceVUS and MCU examinations in 31 patients (62 pelvi-ureter units). Perfect inter-observer agreement (Cohen's kappa statistics = 1.0, p < 0.001) was achieved in ceVUS, suggesting its good reliability in reflux detection and grading. Using MCU as reference, ceVUS had 100 % sensitivity and 84 % specificity and carried higher reflux detection rate than MCU (p < 0.001). There was no complication encountered. Conclusion: Voiding urosonography is a reliable, sensitive, safe and radiation-free modality in the investigation of vesicoureteric reflux in children. It should be incorporated in the diagnostic algorithm in paediatric urinary tract infection.

Scrotal antegradesclerotherapy demonstrates anatomical variations on venous drainage in paediatric, adolescent and young adult varicoceles.

AIM OF THE STUDY: Anatomical variations on venous drainage in varicoceles are under-reported. We report our experience in scrotal antegrade sclerotherapy (SAS) for adolescent varicoceles. METHODS: Since 2011, 15 consecutive boys with left varicoceles were recruited. Under general anaesthesia, a 5-mm transverse incision was made at scrotal neck, testicular vein was cannulated at pampiniform plexus with venogram performed. Foam sclerosant by mixing sodium tetradecyl sulphate (STS), Lipiodol(®) and air was slowly injected under fluoroscopy. Postoperatively the patients were followed-up for varicocele grading, testicular size, and complications. MAIN RESULTS: Median age at operation was 14 (10-19) years. 80 % had grade three varicoceles, 33.3 % had smaller left testis before operation. Intra-operative venogram showed three different anatomical variations. Group I: eleven patients (73.3 %) had single distinct internal spermatic vein; Group II: two patients demonstrated duplication of internal spermatic vein draining into left renal vein; Group III: two patients had pampiniform plexus draining to iliac and/or paraspinal veins. SAS was performed in Group I and II patients. Sclerosant volume injected ranged from 1.5 to 4.5 ml. In Group III patients, surgical ligation of testicular veins was performed rather than SAS to avoid uncontrolled systemic sclerosant spillage. Mean length of stay was 1.13 day. One patient with scrotal haematoma and one other with minor wound dehiscence were managed conservatively. Mean follow-up period was 10.9 (1-22) months. Thirteen patients (86.7 %) achieved varicocele grading ≤ 1. There was no postoperative testicular atrophy, hydrocele and epididymo-orchitis. CONCLUSION: Scrotal antegrade sclerotherapy using STS foam is a safe and effective treatment for adolescent varicoceles. Anatomical variations on venous drainage in varicoceles are common.

Magnetic resonance imaging assessment of substantia nigral iron deposition in Parkinson's disease: a meta-analysis.

OBJECTIVE: The pathogenesis of Parkinson's disease (PD) is associated with abnormal iron accumulation. Magnetic resonance imaging (MRI) studies have shown that patients with Parkinson's disease have an increased amount of iron in their substantia nigra (SN). We have undertaken a meta-analysis of studies using MRI in PD, to explore the potential role of MRI in diagnosing PD using abnormal iron deposition in SN as a candidate biomarker. MATERIALS AND METHODS: Searches of PubMed, Embase, and Medline databases revealed 16 studies that compared PD patients and healthy controls (HC). A sensitivity analysis and subgroup analysis were performed to evaluate the reliability of our results. Estimates were pooled by the fixed-effects model. As an expression of I2, we computed the proportion of variation due to heterogeneity. RESULTS: We included 16 studies with sample sizes of 435 PD and 355 HC in our meta-analysis. Results showed that SN iron deposition was significantly elevated (p<0.00001) in patients with PD compared to HC ones (SMD=0.72, 95% confidence interval 0.57 to 0.87, p<0.00001). CONCLUSIONS: Our findings, based on a homogeneous group-level analysis, suggest that MRI-based SN iron deposition could be used to distinguish PD from HC. For a more rigorous investigation of SN iron deposition in PD, larger cohort studies are needed.

Enzyme replacement therapy for mucopolysaccharidosis VI (Maroteaux-Lamy syndrome): experience in Hong Kong.

Mucopolysaccharidosis VI (Maroteaux-Lamy syndrome) is a very rare inherited lysosomal storage disease. We evaluated the efficacy and safety of weekly infusions of recombinant human arylsulfatase B as enzyme replacement therapy for two patients in whom this condition was advanced. The primary outcome variables were the distance walked in a 6-minute walk test, forced vital capacity, and ejection fraction. The secondary outcome variables were the number of stairs climbed in a 3-minute stair climbing test, joint mobility, urinary glycosaminoglycan excretion, auto-continuous positive airway pressure study and liver size. After 24 weeks of treatment, patient A walked 40 m (36%) and patient B walked 66 m (58%) more in the walk test than at baseline. After 48 weeks, in patient A the corresponding improvements were 142 m (129%) in the walk test and 33 stairs (60%) in the 3-minute stair climbing test, and in patient B the respective improvements were 198 m (174%) and 77 stairs (140%). There was a significant decline in urinary glycosaminoglycan excretion and improvement in range of motion of joints in both patients. The auto-continuous positive airway pressure study revealed improvements in patient A, while other efficacy variables remained static. There were no drug-related adverse events or allergic reactions reported during and after the infusions of recombinant human arylsulfatase B. Recombinant human arylsulfatase B significantly improves endurance and reduces urinary glycosaminoglycan excretion. The drug is generally safe and well tolerated.

[Chi-liang Kwei and an unfulfilled plan of establishing a modern psychiatric hospital in China].

Church General Hospital in Wuchang, China planned to establish a psychiatric hospital in 1930. Chi-liang Kwei, received his PhD at Johns Hopkins Medical School of America and was appointed to be in charge of the preparation. After working out a draft plan for the hospital, Dr. Kwei applied for the Henderson Research Scholarship on Mental Diseases. She planned to study under the guidance of Dr. D.K. Henderson in the Glasgow Mental Royal Hospital for a full year in preparation for the establishment of the hospital. By drawing on the correspondence files of Chi-liang Kwei and psychiatrists Adolf Meyer and David Kennedy Henderson, the paper describes the background conditions and planning for the hospital establishment and interprets the significance and value of Chi-liang Kwei's efforts to establish a psychiatric hospital from the perspective of the historic development of modern psychiatry in China.

Emergence and recovery response of phosphate metabolites and intracellular pH in intact Mytilus edulis as examined in situ by in vivo 31P-NMR.

We employed surface probe-localized 31P-NMR spectroscopy to examine in situ the impact of short-term emergence (hypoxia) and resubmergence on phosphate metabolites and intracellular pH (pHi) in intact mussels. The use of intact organisms ensured that all intrinsic responses remained active while monitoring of individuals minimized uncertainties resulting from stochastic behavior and other individual differences. The use of a photoetched, balanced-match foil probe combined with 1H-NMR images allowed 31P-NMR spectra to be acquired from the posterior adductor muscle with good signal-to-noise. Upon emergence, all mussels exhibited an increase in [Pi], a decline in [phosphoarginine] and pHi, and very little changes in [ATP] with time. The complementary behavior of [phosphoarginine] and [Pi] indicated a precursor-product relationship involved in the maintenance of [ATP] but the similarity between [phosphoarginine] and pHi time-courses cannot be so readily explained. Irregularity in the time-courses of some parameters could have reflected stochastic gaping activity. Resubmergence responses exhibited a reversal of the emergence responses, except that the pHi eventually became supraalkaline with irregular fluctuations. This might be related to the 'oxygen debt' phenomenon and increased oxidative phosphorylation.

Combined use of 1H-NMR and GC-MS for metabolite monitoring and in vivo 1H-NMR assignments.

Thirty-three metabolites were observed in perchloric acid extracts of four different tissues by in vitro 1H-NMR, GC-MS and alcohol dehydrogenase assay, and the information was used to interpret an in vivo two-dimensional nuclear Overhauser effect 1H-NMR spectrum. The metabolite profiles of the different tissues indicate a number of potential tissue-specific markers: N-acetylaspartate and gamma-aminobutyric acid for rat brain, glutamine/glutamic acid ratio for dog heart, arginine and sucrose for carrot, and t-aconitate, sucrose, asparagine/aspartic acid concentration ratios for corn roots. gamma-Aminobutyric acid and malate can be regarded as metabolic indicators for stressed corn roots. Concentrations of threonine and valine in corn roots were constant under hypoxic and salt stress, and can serve as internal standards for both in vivo and in vitro NMR studies. The in vitro information was further used to identify 12 compounds from the in vivo 1H-NMR spectra (including the two-dimensional nuclear Overhauser effect spectrum) of a carrot cylinder by correlating the chemical shift and nuclear Overhauser effect information. Thus, our choice of methods with a capability for structural determination allows the characterization of complex tissue extracts with minimum sample preparation, and supports, as well as complements, in vivo 1H-NMR investigations of metabolism.

Temperature dependence of arginine kinase reaction in the tail muscle of live Sycionia ingentis as measured in vivo by 31P-NMR driven saturation transfer.

We have employed the driven 31P-NMR saturation transfer method to measure in vivo the temperature dependence of the forward and reverse unidirectional fluxes of the arginine kinase reaction in the tail muscle of a live shrimp, Sycionia ingentis. Our results indicated that neither the forward nor the reverse rate constants of this reaction were significantly temperature-dependent between 8 and 16 degrees C, in contrast to the kinetic characteristics of isolated arginine kinases.

Comprehensive chemical profiling of gramineous plant root exudates using high-resolution NMR and MS.

Root exudates released into soil have important functions in mobilizing metal micronutrients and for causing selective enrichment of plant beneficial soil micro-organisms that colonize the rhizosphere. Analysis of plant root exudates typically has involved chromatographic methods that rely on a priori knowledge of which compounds might be present. In the research reported here, the combination of multinuclear and 2-D NMR with GC-MS and high-resolution MS provided de novo identification of a number of components directly in crude root exudates of different plant types. This approach was applied to examine the role of exudate metal ion ligands (MIL) in the acquisition of Cd and transition metals by barley and wheat. The exudation of mugineic acids and malate was enhanced by Fe deficiency. which in turn led to an increase in the tissue content of Cu, Mn, and Zn. The presence of elevated Cd maintained at a free activity pCd of 8.8 (10(-8.8) M), resulted in reduced phytosiderophore production by Fe deficient plants. The buffer morpholinoethane sulfonate (MES), which is commonly used in chelator-buffering nutrient solutions, was detected in the root exudate mixture, suggesting uptake and re-secretion of this compound by the roots. The ability to detect this compound in complex mixtures containing organic acids, amino acids, and other substances suggests that the analytical methods used here provide an unbiased method for simultaneous detection of all major components contained in root exudates.

Phytosiderophores influence on cadmium mobilization and uptake by wheat and barley plants.

A constant anthropogenic release of cadmium to the environment has resulted in a continuous buildup of Cd in soils. Uptake and accumulation of Cd in plant tissue and in grains may lead to food chain transfer to humans. Application of synthetic chelates was suggested to increase metal mobilization and facilitate phytoextraction as a means for the remediation of metal-polluted soils. However, most of the chelate-extracted metal may be leached rather than mobilized to plant roots. In contrast to the synthetic chelates added to soils, plant-produced chelators called phytosiderophores (PS) are excreted directly to the rhizosphere. Previous studies have shown that PS facilitate uptake of Zn and Fe by graminaceous plants. In this study, a two-step PS mediation of Cd uptake was hypothesized: (i) extraction and chelation in the soil solution, and (ii) delivery of the chelated Cd to the uptake system of the plant. We examined Cd extraction by PS, the synthetic chelate HEDTA [N-(2-hydroxyethyl)-ethylenediaminetriacetic acid], and a fungal siderophore rhizoferrin from solid-phase Cd phosphate at pH 7.3 with and without Fe competition in the presence of Ca and Mg as additional competing metals. While rhizoferrin did not extract Cd, PS and HEDTA did extract Cd even in the presence of Fe. Yet, uptake of Cd by wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) plants was not significantly influenced by Fe stress, but instead was controlled primarily by Cd2+ activity in solution. These results suggest that even though Cd may be mobilized by PS, there is no significant uptake of the Cd-PS complex by the plant roots.

Relationships between sediment microbial communities and pollutants in two California salt marshes.

Salt marshes are important ecosystems whose plant and microbial communities can alter terrestrially derived pollutants prior to coastal water discharge. However, knowledge regarding relationships between anthropogenic pollutant levels and salt marsh microbial communities is limited, and salt marshes on the West Coast of the United States are rarely examined. In this study, we investigated the relationships between microbial community composition and 24 pollutants (20 metals and 4 organics) in two California salt marshes. Multivariate ordination techniques were used to assess how bacterial community composition, as determined by terminal restriction fragment length polymorphism and phospholipid fatty acid analyses, was related to pollution. Sea urchin embryo toxicity measurements and plant tissue metabolite profiles were considered two other biometrics of pollution. Spatial effects were strongly manifested across marshes and across channel elevations within marshes. Utilizing partial canonical correspondence analysis, an ordination technique new to microbial ecology, we found that several metals were strongly associated with microbial community composition after accounting for spatial effects. The major patterns in plant metabolite profiles were consistent with patterns across microbial community profiles, but sea urchin embryo assays, which are commonly used to evaluate ecological toxicity, had no identifiable relationships with pollution. Whereas salt marshes are generally dynamic and complex habitats, microbial communities in these marshes appear to be relatively sensitive indicators of toxic pollutants.

Identification of glycerophosphorylcholine in mussel ovarian extracts by two-dimensional nuclear magnetic resonance.

The abundance of the "phosphodiester" peak in differentiating or proliferating tissues, including reproductive organs and tumors, warrants further investigations of its metabolic role(s), which would require a rigorous confirmation of its identity. The assignment of this peak to glycerophosphorylcholine in 31P NMR spectra of biological samples has been largely based on chemical shift, which can result in ambiguities. We employed a combination of two-dimensional 31P-1H heteronuclear shift correlation and 1H total correlation spectroscopies to trace the spin connectivities of glycerophosphorylcholine and thus to identify its structure directly from crude ovarian extracts of mussels without ambiguities and the need for extensive purification. This approach can be applied generally to the identification of molecules containing heteroatoms in crude tissue extracts.

Isolation and characterization of two cyanogenic beta-glucosidases from flax seeds.

Two cyanogenic beta-glucosidases, linustatinase and linamarase, were isolated and purified from flax seeds (Linum ussitatissimum). They catalyze the sequential hydrolysis of linustatin and neolinustatin to yield acetone and methylethyl ketone cyanohydrins, respectively. The purification procedure for linustatinase involved acetone extraction, precipitation by polyethyleneimine and ammonium sulfate (40-80% saturation), and Red A gel, concanavalin A-Sepharose, and PBE 94 column chromatography; that for linamarase was similar except that polyethyleneimine precipitation was eliminated and DE-52 and Sepharose CL-6B replaced Red A gel column chromatography. The native substrates neolinustatin and linamarin were used for the assay during purification. Both proteins were purified to electrophoretic homogeneity. Linustatinase is an alpha beta dimer (molecular weights of alpha and beta = 39,000 and 19,000, respectively) while linamarase appears to be an alpha 5 beta 5 decamer (molecular weights of alpha and beta = 62,500 and 65,000, respectively). Both enzymes contain mannose or glucose. Linustatinase exists in five different isozymic forms (isoelectric points between 7 and 8) whereas linamarase occurs in one major form (isoelectric point 4 to 5). The kinetic parameters of the two enzymes are similar: acidic pH optima, Km's in the millimolar range, and competitive inhibition by delta-gluconolactone, a transition state analog. The presence of an aglycone structure in the substrates is important for both enzyme activities. In addition, both enzymes are specific towards the beta-glycosidic linkage; linustatinase (a beta-bis-glucosidase) readily hydrolyzes beta-bis-glucosides with 1,6 and 1,3 linkages whereas linamarase (a beta-monoglucosidase) exhibits little activity towards these substrates.

In vivo 13C NMR analysis of acyl chain composition and organization of perirenal triacylglycerides in rats fed vegetable and fish oils.

Lipid composition of body fat can be a key indicator of nutritional status and a number of human disorders. In vivo 13C NMR provides for repeated, noninvasive analysis of fatty acyl chain composition on individuals, which circumvents classical problems of individual variation and repetitive invasive sampling. It also offers a unique opportunity to examine acyl chain organization in situ. This approach was used to examine the fatty acyl chain composition in the perirenal fat pads of rats fed olive, safflower, and menhaden oil-containing diets. These changes were then monitored during a diet switch between olive and menhaden oil-fed rats. The fatty acid composition of perirenal fat pads and livers was also analyzed using gas chromatography for comparison with the in vivo NMR analysis. Both tissues assumed the general characteristics of diet fatty acyl chain and fatty acid composition and the diet switch induced a switchover of the perirenal composition in 30-45 days. These results indicate that a large portion of the diet fatty acyl chains were incorporated directly into adipose and liver tissues although some were also metabolized, particularly in menhaden oil-fed rats. Furthermore, changes in the in vivo spin-lattice relaxation times (T1) of fatty acyl carbons in the perirenal fat pads and their lipid extracts were followed and effective correlation times (tau eff) were calculated from the T1 data. The result indicated that the in vivo segmental mobility of acyl carbons was sensitive to changes in diet-derived fatty acyl chain composition and that the central region of the acyl chain was more sensitive to these changes. There was a qualitative similarity but quantitative differences in the tau eff of acyl carbons acquired in vivo and from extracts. These results suggest that adipose triacylglycerides experience an overall liquid-like microenvironment in vivo but with more restriction in their mobility, and that different factors may exist in governing their organization in situ versus in extracts.

Monitoring of hypoxic metabolism in superfused plant tissues by in vivo 1H NMR.

We have used a coaxial superfusion system to obtain physiologically interpretable in vivo 1H NMR spectra at 500 MHz of carrot roots, maize roots, and rice shoots in water (no 2H2O). The superfusion system was constructed from common laboratory parts, required no modification of the probe and sample loading procedure, and was inherently leak resistant. The assignment and quantitation of the in vivo 1H NMR resonances were achieved by performing two-dimensional NMR experiments in vivo, and by in vitro analysis including NMR and gas chromatography-mass spectrometry. The in vivo spectra were dominated by resonances arising from sugars, organic acids, amino acids, and ethanol. In vivo measurements of spin-lattice relaxation times and chemical shifts of beta protons of malate in carrot roots suggested that malate was located in a relatively viscous and acidic compartment. In rice shoots, the hypoxic time courses of 9 metabolites were established in vivo, and 23 in vitro. In both cases, accumulation of lactate, ethanol, Ala, and gamma-aminobutyrate as well as a decrease in Gln and Asn concentrations were observed. These findings are consistent with accelerated glycolysis and decreased tricarboxylic acid cycle activity.

Hypoxia does not affect rate of ATP synthesis and energy metabolism in rice shoot tips as measured by 31P NMR in vivo.

The cytoplasmic pH, concentrations of phosphate metabolites, and rate of ATP synthesis were measured in vivo in excised rice shoot tips under normoxic and hypoxic conditions using 31P NMR. When supplied with glucose, the shoot tips grew rapidly and were relatively unaffected by hypoxia. The cytoplasmic pH decreased transiently by only 0.2 units during hypoxia, and the concentration of ATP was maintained to at least 90% of the normoxic level. Most importantly, the unidirectional rate constant of ATP synthesis from free phosphate decreased less than 25% during hypoxia. This is in contrast to other actively growing tissues such as the maize root tip. gamma-Aminobutyrate was the major nonvolatile fermentation end product after 22 h of hypoxia. Other hypoxia-induced changes included a modest increase in [Ala] and [succinate] as well as a substantial decrease in [malate].

An in vivo 1H and 31P NMR investigation of the effect of nitrate on hypoxic metabolism in maize roots.

The effect of nitrate on the short-term hypoxic response and recovery of flooded mature maize roots has been investigated in vivo by 1H and 31P NMR and in vitro by 1H NMR and gas chromatography-mass spectrometry. Employing 1H NMR in addition to 31P NMR extended the number of identifiable compounds in vivo from 4 to 15, while in vitro two-dimensional NMR and gas chromatography-mass spectrometry aided rigorous in vivo 1H NMR resonance assignments and quantitation of 24 compounds. In the absence of nitrate, the concentrations of key metabolites including alanine, ethanol, gamma-aminobutyrate, lactate, succinate, and sucrose changed during 8 h of hypoxia in a manner consistent with reduced tricarboxylic acid cycle activity and diversion to glycolytic fermentation. The pH drop in the cytoplasm during hypoxia was rapid, about 0.2 unit, and diminished quickly upon recovery. Rapid recovery of ethanol, succinate, and sucrose levels was also observed, which indicates a return to normal aerobic metabolism. Although the hypoxic response itself, including pH, was not greatly affected by the presence of nitrate, nitrate reduced the amount of fermentation end products produced, helped maintain a higher free NTP concentration during hypoxia, and increased the rate of overall recovery from hypoxia. These findings suggest the presence of a nitrate-induced maintenance-level respiration in hypoxic maize roots, which helps explain the protection imparted by nitrate to flooded hypoxic maize plants.

Monitoring of metabolic responses of intact Haliotis (abalones) under salinity stress by 31P surface probe localized NMR.

Surface probe localized 31P NMR spectroscopy was employed to record the metabolic responses of the foot of intact Haliotis cracherodii and H. rufescens (black and red abalones) under hyper- and hypoosmotic stresses. Use of the surface probe allowed spectral localization on the foot of intact abalones, facilitated monitoring of different sizes of animals, and minimized constraints on aquatic chamber design normally imposed by homogeneous-field probes. Generally, hyperosmotic stress (51%) elicited more rapid changes of phosphate metabolites than hypoosmotic stress (17%). As with the well-studied hypoxic stress in intact mammalian and excised molluscan tissue, both salinity treatments caused drops in the phosphagen and increases in inorganic phosphate levels. However, osmotic stress was distinct from hypoxic stress in that intracellular pH did not change and nucleotide triphosphate (NTP) concentrations dropped immediately. Although these findings are preliminary, they demonstrate the utility of the surface probe approach for studies of environmental stress in intact marine invertebrates.