Current status of laboratory and imaging diagnosis of neonatal necrotizing enterocolitis

Necrotizing enterocolitis continues to be a devastating disease process for very low birth weight infants in Neonatal Intensive Care Units. The aetiology and pathogenesis of necrotizing enterocolitis are not definitively understood. It is known that necrotizing enterocolitis is secondary to a complex interaction of multiple factors that results in mucosal damage, which leads to intestinal ischemia and necrosis. Advances in neonatal care, including resuscitation and ventilation support technology, have seen increased survival rates among premature neonates and a concomitant detection in the incidence of this intestinal disease.Diagnosis can be difficult, and identifying infants at the onset of disease remains a challenge. Early diagnosis, which relies on imaging findings, and initiation of prompt therapy are essential to limit morbidity and mortality. Moreover, early management is critical and life-saving.This review summarizes what is known on the laboratory and instrumental diagnostic strategies needed to improve neonatal outcomes and, possibily, to prevent the onset of an overt necrotizing enterocolitis

Evaluation of soil-microbial communities by their CLPP. Standardization of a laboratory technique to replace commercial available microplates

La variación de la composición de algunas comunidades microbianas edáficas son buenos bioindicadores del impacto de la actividad antrópica sobre los suelos, tales como diferentes formas de manejo o su contaminación. Los métodos tradicionales de aislamiento y análisis taxonómico no consideran la funcionalidad de las comunidades microbianas, por lo que los perfiles fisiológicos de uso de fuentes carbonadas (CLPP) constituyen una metodología complementaria para su estudio. Numerosos trabajos demostraron que las microplacas de Biolog® EcoPlates® son útiles para determinar diferencias fisiológicas entre comunidades de diferentes suelos. Sin embargo, estas microplacas comerciales poseen algunas desventajas, por lo cual surgió la idea de reemplazarlas por microplacas preparadas en el laboratorio. Comparamos ambos tipos de microplacas con muestras de suelo provenientes de un ensayo de biorremediación. Analizamos a) el desarrollo promedio de color para cada tratamiento, b) los valores promedio de absorbancia para cada tipo de microplaca, c) los análisis de componentes principales, y d) el índice de diversidad de Shannon-Weaver (H) para cada muestra. Si bien los valores promedio de absorbancia difirieron significativamente entre ambos tipos de microplacas, los resultados del análisis de componentes principales y de diversidad fueron relativamente similares. En conclusión, ambos tipos de microplacas resultaron similares para detectar diferencias en los CLPP de los distintos tratamientos. Es por ello que las microplacas preparadas en el laboratorio constituyen una herramienta confiable y económica para el estudio de la fisiología de comunidades microbianas de suelo.Variation of soil-microbial communities are good bioindicators of human impacts in soils, such as different soils management or contamination. Considering that traditional methods of isolation and taxonomic analysis do not consider the functionality of the microbial community, Community-Level Physiological Profiles (CLPP) emerged as a complementary methodology to study microbial communities. Several studies have shown that Biolog® EcoPlates® are very useful for determining physiological differences between communities from different samples. However, commercial microplates have some disadvantages which led us to the idea of replacing them by microplates prepared in the laboratory (Laboratoryms). Here, we compared both types of microplates using soil samples from a bioremediation assay. We compared a) the average well color development for each sample, b) the averages of absorbance values for each type of microplate, c) Principal Components, and d) Shannon-Weaverms diversity index (H). Although Laboratoryms showed significantly lower Average absorbance values than EcoPlates®, the principal component analysis and diversity index did not differ between types of microplates. In conclusion, both types of microplates showed a relatively similar ability to detect differences in the CLPP of the treatments studied. Consequently, microplates prepared in laboratory are a reliable and economical tool to study the physiology of soil microbial communities

Evaluación de comunidades microbianas edáficas mediante CLPP : estandarización de una técnica de laboratorio para reemplazar microplacas comerciales

129-136Variation of soil-microbial communities are good bioindicators of human impacts in soils, such as different soils management or contamination. Considering that traditional methods of isolation and taxonomic analysis do not consider the functionality of the microbial community, Community-Level Physiological Profiles (CLPP) emerged as a complementary methodology to study microbial communities. Several studies have shown that Biolog® EcoPlates® are very useful for determining physiological differences between communities from different samples. However, commercial microplates have some disadvantages which led us to the idea of replacing them by microplates prepared in the laboratory (Laboratory's). Here, we compared both types of microplates using soil samples from a bioremediation assay. We compared a) the average well color development for each sample, b) the averages of absorbance values for each type of microplate, c) Principal Components, and d) Shannon-Weaver's diversity index (H). Although Laboratory's showed significantly lower Average absorbance values than EcoPlates®, the principal component analysis and diversity index did not differ between types of microplates. In conclusion, both types of microplates showed a relatively similar ability to detect differences in the CLPP of the treatments studied. Consequently, microplates prepared in laboratory are a reliable and economical tool to study the physiology of soil microbial communities

Evaluation of an Internet Document Delivery Service

An Internet-based Document Delivery Service (DDS) has been developed within the framework of the CNR ( the Italian Research National Council) Project BiblioMIME, in order to take advantage of new Internet technologies and promote cooperation among CNR and Italian university libraries. Adopting such technologies changes the traditional organisation of DDS and may drastically reduce costs and delivery times. An information system managing DDS requests and monitoring the temporal evolution of the service has been implemented, running on the local-area network of a test-site library. It aims to track number and types of documents requested and received, user distribution, delivery times and types (surface mail, fax, Internet), to automate repetitive manual procedures and to deal with the various accounting methods used by other libraries. Transmission of documents is carried out by means of an e-mail/Web gateway system supporting document exchange via Internet, which assists receiving libraries in retrieving requested documents. This paper describes the architecture and main design features of the e-mail/Web gateway server (the BiblioMime server). This approach permits librarians to continue using e-mail service to send large documents, while resolving problems that users may encounter when downloading large size files with e-mail agents. The library operator sends the document as an attachment to the destination address; on fly the e-mail server extracts and saves the attachments in a web-server disk file and substitutes them with a new message part that includes an URL pointing to the saved document. The receiver can download these large objects by means of a user-friendly browser. We further discuss the data gathered during the triennium 1998-2000; this consists of about 5,000 DDS transactions per annum with 300 other Italian scientific and bio-medical libraries and commercial document suppliers. Use of the instruments described above allowed us to evaluate the performance of service "before" and "after" the use of Internet Document Delivery and to extract some critical data regarding DDS. Those include: a) libraries with which we have greater numbers of exchanges and their turnaround times; b) extraordinary reduction in costs and delivery times; c) the most frequently requested serial titles (allowing cost-effective decisions on new subscriptions); d) impact on DDS of library participation in consortia which allow user access to greater numbers of online serials

Validation of an electrogoniometry system as a measure of knee kinematics during activities of daily living

Purpose: The increasing use of electrogoniometry (ELG) in clinical research requires the validation of different instrumentation. The purpose of this investigation was to examine the concurrent validity of an ELG system during activities of daily living. Methods: Ten asymptomatic participants gave informed consent to participate. A Biometrics SG150 electrogoniometer was directly compared to a 12 camera three dimensional motion analysis system during walking, stair ascent, stair descent, sit to stand, and stand to sit activities for the measurement of the right knee angle. Analysis of validity was undertaken by linear regression. Standard error of estimate (SEE), standardised SEE (SSEE), and Pearson’s correlation coefficient r were computed for paired trials between systems for each functional activity. Results: The 95% confidence interval of SEE was reasonable between systems across walking (LCI = 2.43 °; UCI = 2.91 °), stair ascent (LCI = 2.09 °; UCI = 2.42 °), stair descent (LCI = 1.79 °; UCI = 2.10 °), sit to stand (LCI = 1.22 °; UCI = 1.41 °), and stand to sit (LCI = 1.17 °; UCI = 1.34 °). Pearson’s correlation coefficient r across walking (LCI = 0.983; UCI = 0.990), stair ascent (LCI = 0.995; UCI = 0.997), stair descent (LCI = 0.995; UCI = 0.997), sit to stand (LCI = 0.998; UCI = 0.999), and stand to sit (LCI = 0.996; UCI = 0.997) was indicative of a strong linear relationship between systems. Conclusion: ELG is a valid method of measuring the knee angle during activities representative of daily living. The range is within that suggested to be acceptable for the clinical evaluation of patients with musculoskeletal conditions

Evaluación de comunidades microbianas edáficas mediante CLPP : estandarización de una técnica de laboratorio para reemplazar microplacas comerciales

129-136Variation of soil-microbial communities are good bioindicators of human impacts in soils, such as different soils management or contamination. Considering that traditional methods of isolation and taxonomic analysis do not consider the functionality of the microbial community, Community-Level Physiological Profiles (CLPP) emerged as a complementary methodology to study microbial communities. Several studies have shown that Biolog® EcoPlates® are very useful for determining physiological differences between communities from different samples. However, commercial microplates have some disadvantages which led us to the idea of replacing them by microplates prepared in the laboratory (Laboratory's). Here, we compared both types of microplates using soil samples from a bioremediation assay. We compared a) the average well color development for each sample, b) the averages of absorbance values for each type of microplate, c) Principal Components, and d) Shannon-Weaver's diversity index (H). Although Laboratory's showed significantly lower Average absorbance values than EcoPlates®, the principal component analysis and diversity index did not differ between types of microplates. In conclusion, both types of microplates showed a relatively similar ability to detect differences in the CLPP of the treatments studied. Consequently, microplates prepared in laboratory are a reliable and economical tool to study the physiology of soil microbial communities

A modified culture medium for improved isolation of marine vibrios

Marine Vibrio members are of great interest for both ecological and biotechnological research, which often relies on their isolation. Whereas many efforts have been made for the detection of food-borne pathogenic species, much less is known about the performances of standard culture media toward environmental vibrios. We show that the isolation/enumeration of marine vibrios using thiosulfate-citrate-bile salts-sucrose agar (TCBS) as selective medium may be hampered by the variable adaptability of different taxa to the medium, which may result even in isolation failure and/or in substantial total count underestimation. We propose a modified TCBS as isolation medium, adjusted for marine vibrios requirements, which greatly improved their recovery in dilution plate counts, compared with the standard medium. The modified medium offers substantial advantages over TCBS, providing more accurate and likely estimations of the actual presence of vibrios. Modified TCBS allowed the recovery of otherwise undetected vibrios, some of which producing biotechnologically valuable enzymes, thus expanding the isolation power toward potentially new enzyme-producers Vibrio taxa. Moreover, we report a newly designed Vibrio-specific PCR primers pair, targeting a unique rpoD sequence, useful for rapid confirmation of isolates as Vibrio members and subsequent genetic analyses

Efficiency of sperm-mediated gene transfer in the ovine by laparoscopic insemination, in vitro fertilization and ICSI

188-196Transgenesis constitutes an important tool for pharmacological protein production and livestock improvement. We evaluated the potential of laparoscopic insemination (LI), in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) to produce egfp-expressing ovine embryos, using spermatozoa previously exposed to pCX-EGFP plasmid in two different sperm/DNA incubation treatments: "Long Incubation" (2 h at 17 C) and "Short Incubation" (5 min at 5 C). For LI, Merino sheep were superovulated and inseminated with treated fresh semen from Merino rams. The embryos were recovered by flushing the uterine horns. For IVF and ICSI, slaughterhouse oocytes were fertilized with DNA-treated frozen/thawed sperm. All recovered embryos were exposed to blue light (488 nm) to determine green fluorescent morulae and blastocysts rates. High cleavage and morulae/blastocysts rates accompanied the LI and IVF procedures, but no egfp-expressing embryos resulted. In contrast, regardless of the sperm/ plasmid incubation treatment, egfp-expressing morulae and blastocysts were always obtained by ICSI, and the highest transgenesis rate (91.6 percent) was achieved with Short Incubation. In addition, following the incubation of labeled plasmid DNA, after Long or Short exposure treatments, with fresh or frozen/thawed spermatozoa, only non-motile fresh spermatozoa could maintain an attached plasmid after washing procedures. No amplification product could be detected following PCR treatment of LI embryos whose zonae pellucidae (ZP) had been removed. In order to establish conditions for transgenic ICSI in the ovine, we compared three different activation treatments, and over 60 percent of the obtained blastocysts expressed the transgene. For ICSI embryos, FISH analysis found possible signals compatible with integration events. In conclusion, our results show that in the ovine, under the conditions studied, ICSI is the only method capable of producing exogenous gene-expressing embryos using spermatozoa as vectors

Profiles of Urine Samples Taken from Ecstasy Users at Rave Parties: Analysis by Immunoassays, HPLC, and GC-MS

The abuse of the designer amphetamines such as 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy) is increasing throughout the world. They have become popular drugs, especially at all-night techno dance parties (Raves), and their detection is becoming an important issue. Presently, there are no MDMA- or MDA-specific immunoassays on the market, and detection of the designer amphetamines is dependent upon the use of commercially available amphetamine assays. The success of this approach has been difficult to assess because of the general unavailability of significant numbers of samples from known drug users. The objectives of the present study are to characterize the drug content of urine samples from admitted Ecstasy users by chromatographic methods and to assess the ability of the available amphetamine/methamphetamine immunoassays to detect methylenedioxyamphetamines. We found that, when analyzed by high-performance liquid chromatography with diode-array detection (HPLC-DAD), 64% of 70 urine samples (by gas chromatography-mass spectrometry [GC-MS]: 88% of 64 urine samples) obtained from Rave attendees contained MDMA and/or 3,4-methylenedioxyamphetamine (MDA) alone or in combination with amphetamine, methamphetamine, or other designer amphetamines such as 3,4-methylenedioxyethylamphetamine (MDEA). This suggests that the majority of the Ravers are multi-drug users. At the manufacturer's suggested cutoffs, the Abbott TDx Amphetamine/Methamphetamine II and the new Roche HS Amphetamine/MDMA assays demonstrated greater detection sensitivity for MDMA than the other amphetamine immunoassays tested (Abuscreen OnLine Hitachi AMPS, Abuscreen OnLine Integra AMPS, Abuscreen OnLine Integra AMPSX, CEDIA AMPS, and EMIT II AMPS). There is 100% agreement between each of the two immunoassays with the reference chromatographic methods, HPLC-DAD and GC-MS, for the detection of methylenedioxyamphetamine

Antioxidant activities of hydroxylated naphthalenes: the role of aryloxyl radicals

Herein is delineated a first systematic framework for the definition of structure-antioxidant property relationships in the dihydroxynaphthalene (DHN) series. The results obtained by a combined experimental and theoretical approach revealed that 1,8-DHN is the best performing antioxidant platform, with its unique hydrogen-bonded peri-hydroxylation pattern contributing to a fast H atom transfer process. Moreover, the comparative analysis of the antioxidant properties of DHNs carried out by performing DPPH and FRAP assays and laser flash photolysis experiments, revealed the higher antioxidant power associated with an α-substitution pattern (i. e. in 1,8- and 1,6-DHN) with respect to DHNs exhibiting a β-substitution pattern (i. e. in 2,6- and 2,7-DHN). DFT calculations and isolation and characterization of the main oligomer intermediates formed during the oxidative polymerization of DHNs supported this evidence by providing unprecedented insight into the generation and fate of the intermediate naphthoxyl radicals, which emerged as the main factor governing the antioxidant activity of DHNs