Enhanced and green extraction of bioactive compounds from Lippia citriodora by tailor-made natural deep eutectic solvents.

Natural deep eutectic solvents (NADESs) have been postulated as alternative green solvents for the isolation of valuable bioactive compounds from Lippia citriodora. Thus, 11 different NADESs, based on choline chloride (ChCl) as the hydrogen bond acceptor in combination with different hydrogen bond donors (organic acids, polyalcohols, sugars, and urea) were tested. According to the results obtained, ChCl-lactic acid exhibited the highest extraction yield for iridoids, 7.25 mg g-1, phenylpropanoids, 17.23 mg g-1, and flavonoids, 9.02 mg g-1 being significantly greater than phenylpropanoid and flavonoid yields, 15.63 and 5.43 mg g-1 respectively, extracted with methanol as conventional solvent. Subsequently, in order to optimise the most influential microwave assisted extraction (MAE) parameters, a Box-Behnken design paired with a response surface methodology were implemented. Temperature and water content showed a strong effect on the extraction of polyphenol sub-classes, while the effect of irradiation time was less noticeable on extraction yields. Temperature of 63.68 °C, a water content of 32.19% and a microwave irrdiation time of 17.08 min were the optimum conditions provided by the statistical program. The use of NADESs showed potential to facilitate the design and customisation of green tailor-made solvents which have greater extraction capacity than conventional organic solvents.

Real-time chest-wall-motion tracking by a single optical fibre grating: a prospective method for ventilator triggering.

OBJECTIVE: The ventilators involved in non-invasive mechanical ventilation commonly provide ventilator support via a facemask. The interface of the mask with a patient promotes air leaks that cause errors in the feedback information provided by a pneumatic sensor and hence patient-ventilator asynchrony with multiple negative consequences. Our objective is to test the possibility of using chest-wall motion measured by an optical fibre-grating sensor as a more accurate non-invasive ventilator triggering mechanism. APPROACH: The basic premise of our approach is that the measurement accuracy can be improved by using a triggering signal that precedes pneumatic triggering in the neuro-ventilatory coupling sequence. We propose a technique that uses the measurement of chest-wall curvature by a long-period fibre-grating sensor. The sensor was applied externally to the rib-cage and interrogated in the lateral (edge) filtering scheme. The study was performed on 34 healthy volunteers. Statistical data analysis of the time lag between the fibre-grating sensor and the reference pneumotachograph was preceded by the removal of the unwanted heartbeat signal by wavelet transform processing. MAIN RESULTS: The results show a consistent fibre-grating signal advance with respect to the standard pneumatic signal by (230 ± 100) ms in both the inspiratory and expiratory phases. We further show that heart activity removal yields a tremendous improvement in sensor accuracy by reducing it from 60 ml to 0.3 ml. SIGNIFICANCE: The results indicate that the proposed measurement technique may lead to a more reliable triggering decision. Its imperviousness to air leaks, non-invasiveness, low-cost and ease of implementation offer good prospects for applications in both clinical and homecare ventilation.

Rib-cage-movement measurements as a potential new trigger signal in non-invasive mechanical ventilation.

Non-invasive ventilation performed through an oronasal mask is a standard in clinical and homecare mechanical ventilation. Besides all its advantages, inevitable leaks through the mask cause errors in the feedback information provided by the airflow sensor and, hence, patient-ventilator asynchrony with multiple negative consequences. Here we investigate a new way to provide a trigger to the ventilator. The method is based on the measurement of rib cage movement at the onset of inspiration and during breathing by fibre-optic sensors. In a series of simultaneous measurements by a long-period fibre grating sensor and pneumotachograph we provide the statistical evidence of the 200 ms lag of the pneumo with respect the fibre-optic signal. The lag is registered consistently across three independent delay metrics. Further, we discuss exceptions from this trend and identify the needed improvements to the proposed fibre-sensing scheme.

Factores de riesgo cardiovascular en estudiantes universitarios / Cardiovascular risk factors of college students

College students are in a key life stage for the adoption of lifestyles that will be practiced in the family, society and work. During this period, students gain greater autonomy and take responsibility for their own care. University time becomes a critical step in the development of lifestyles. There is a high prevalence of risk factors for non-communicable diseases in university students such as pre-hypertension especially in men and hypercholesterolemia in women. Other risk factors include smoking, physical inactivity, overweight, obesity, low fruit and vegetable consumption, and high consumption of saturated fats. Some differences of cardiovascular risk factors have been found according to gender, year of career and faculty where students attend. It is recommended to consider these differences when we design and perform educational interventions to achieve greater assertiveness and effectiveness. This article reviews the evidence about cardiovascular risk factors in university students according to gender, year of career and faculty where students attend, following the model proposed by Cecchini et al and according to the Framingham study.

Los estudiantes universitarios se encuentran en una etapa del ciclo vital clave para la adopción de estilos de vida, que practicarán en el ámbito familiar, social y laboral. En este periodo los estudiantes adquieren mayor autonomía y asumen la responsabilidad de su autocuidado, por lo cual se convierte en una etapa crítica para el desarrollo de sus estilos de vida. Existe una alta prevalencia de factores de riesgo de enfermedades crónicas no transmisibles en los universitarios, entre los que destacan pre-hipertensión especialmente en hombres y la hipercolesterolemia en mujeres. También el tabaquismo, sedentarismo, sobrepeso, obesidad, bajo consumo de frutas y verduras, y alto consumo de grasas saturadas. Se encontraron diferencias en los factores de riesgo cardiovascular de acuerdo al género, curso y facultad de los estudiantes. Se recomienda consideran estas diferencias al diseñar y realizar intervenciones educativas para lograr una mayor asertividad y efectividad. El presente artículo revisa la evidencia sobre los factores de riesgo cardiovascular en estudiantes universitarios de acuerdo al género, curso y facultad, según el modelo propuesto por Cecchini y cols. y al estudio de Framingham.

Effect of Ritonavir on (99m)Technetium-Mebrofenin Disposition in Humans: A Semi-PBPK Modeling and In Vitro Approach to Predict Transporter-Mediated DDIs.

A semiphysiologically based pharmacokinetic (semi-PBPK) model was developed to describe a unique blood, liver, and bile clinical data set for the hepatobiliary imaging agent (99m)Technetium-mebrofenin ((99m)Tc-mebrofenin), and to simulate sites/mechanisms of a (99m)Tc-mebrofenin-ritonavir drug-drug interaction (DDI). The transport inhibitor ritonavir (multiple-dose: 2 × 300 mg) significantly increased systemic (99m)Tc-mebrofenin exposure as compared with control (4,464 ± 1,861 vs. 1,970 ± 311 nCi min/ml; mean ± SD), without affecting overall hepatic exposure or biliary recovery. A novel extrahepatic distribution compartment was required to characterize (99m)Tc-mebrofenin disposition. Ritonavir inhibited (99m)Tc-mebrofenin accumulation in human sandwich-cultured hepatocytes (SCH) (half maximal inhibitory concentration (IC50) = 3.46 ± 1.53 µmol/l). Despite ritonavir accumulation in hepatocytes, intracellular binding was extensive (97. 6%), which limited interactions with multidrug resistance protein 2 (MRP2)-mediated biliary excretion. These in vitro data supported conclusions from modeling/simulation that ritonavir inhibited (99m)Tc-mebrofenin hepatic uptake, but not biliary excretion, at clinically relevant concentrations. This integrated approach, utilizing modeling, clinical, and in vitro data, emphasizes the importance of hepatic and extrahepatic distribution, assessment of inhibitory potential in relevant in vitro systems, and intracellular unbound concentrations to assess transporter-mediated hepatic DDIs.CPT: Pharmacometrics & Systems Pharmacology (2013) 2, e20; doi:10.1038/psp.2012.21; advance online publication 2 January 2013.

First Report of Botryosphaeria dothidea Causing White Rot of Apple Fruit in Serbia.

Botryosphaeria dothidea (Moug.: Fr.) Ces. & De Not has a worldwide distribution infecting species from over 80 genera of plants (1). Apart from being an important pathogen of apple trees in many countries, B. dothidea can cause pre- and postharvest decay on apple fruit (2). It has been known to cause canker and dieback of forest trees in Serbia (3), but has not been recorded either on apple trees or apple fruit. In December 2010, apple fruit cv. Idared (Malus × domestica Borkh.) with symptoms of white rot were collected from one storage in the area of Svilajnac in Serbia. The incidence of the disease was low but the symptoms were severe. Affected fruit were brown, soft, and almost completely decayed, while the internal decayed tissue appeared watery and brown. A fungus was isolated from symptomatic tissue of one fruit after surface sterilization with 70% ethanol (without rinsing) and aseptic removal of the skin. Small fragments of decayed tissue were placed on potato dextrose agar (PDA) and incubated in a chamber at 22°C under alternating light and dark conditions (12/12 h). Fungal colonies were initially whitish, but started turning dark gray to black after 5 to 6 days. Pycnidia were produced after 20 to 25 days of incubation at 22°C and contained one-celled, elliptical, hyaline conidia. Conidia were 17.19 to 23.74 µm (mean 18.93) × 3.72 to 4.93 µm (mean 4.45) (n = 50). These morphological characteristics are in accordance with those described for the fungus B. dothidea (4). Genomic DNA was isolated from the fungus and internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The nucleotide sequence has been assigned to GenBank Accession No. KC994640. BLAST analysis of the 528-bp segment showed a 100% similarity with several sequences of B. dothidea deposited in NCBI GenBank, which confirmed morphological identification. Pathogenicity was tested by wound inoculation of five surface-sterilized, mature apple fruit cv. Idared with mycelium plugs (5 mm in diameter) of the isolate grown on PDA. Five control fruit were inoculated with sterile PDA plugs. After 5 days of incubation in plastic containers, under high humidity (RH 90 to 95%) at 22°C, typical symptoms of white rot developed on inoculated fruit, while wounded, uninoculated, control fruit remained symptomless. The isolate recovered from symptomatic fruit showed the same morphological features as original isolate. To the best of our knowledge, this is the first report of B. dothidea on apple fruit in Serbia. Apple is widely grown in Serbia and it is important to further investigate the presence of this pathogen in apple storage, as well as in orchards since B. dothidea may cause rapid disease outbreaks that result in severe losses. References: (1) G. H. Hapting Agriculture Handbook 386, USDA, Forest Service, 1971. (2) A. L. Jones and H. S. Aldwinckle Compendium of Apple and Pear Diseases. APS Press, St. Paul, MN, 1990. (3) D. Karadzic et al. Glasnik Sumarskog Fakulteta 83:87, 2000. (4) B. Slippers et al. Mycologia 96:83, 2004.

First Report of Brown Rot Caused by Monilia polystroma on Apple in Serbia.

Monilia polystroma van Leeuwen is a new Japanese species, similar to M. fructigena but distinguishable based on morphological and molecular characteristics (3). After its first discovery on apple in Japan, occurance of M. polystroma in Europe has been reported in Hungary, the Czech Republic, and Switzerland (2,3,4). In October 2011, during a survey for apple fungal pathogens in the Bela Crkva district, 15 apple fruit (Malus domestica Borkh.) cv. Golden Delicious were collected. Two isolates of Monilinia polystroma were obtained from apple fruit showing brown rot, covered with small yellowish sporodohia. The pathogen was identified as M. polystroma based on morphological and molecular features (1,3). Upon isolation, colonies cultivated on PDA were white to grayish and the mycelium grew 8.85 mm per day at 22 ± 1°C in 12-h light/12-h dark regime. After 6 to 8 days of incubation, black stromatal plates were observed on the reverse sides of the inoculated petri dishes. Conidia were one-celled, limoniform, hyaline, 14.7 to 21.88 µm (16.2 mean) × 7.85 to 12.92 µm (10.8 mean), and were produced in branched monilioid chains on inoculated apple fruit. Morphological identification was confirmed by PCR (1) using genomic DNA extracted from the mycelium of pure cultures, and amplified products of 425 bp in length, specific for M. polystroma were amplified as expected with primers MO368-5 and MO368-8R. For one isolate, the ribosomal ITS1-5.8S-ITS2 region was obtained, using primers ITS1 and ITS4, and deposited in GenBank (Accession No JX315717). The sequence was 498 bp in length and showed 100% identity with sequences deposited for M. polystroma in NCBI GenBank (JN128835, AM937114, GU067539). Pathogenicity was confirmed by wound-inoculating five surface-sterilized, mature apple fruit with mycelium plugs (5 mm in diameter) of both isolates grown on PDA. Control fruit were inoculated with sterile PDA plugs. After 3 days of incubation in plastic containers, under high humidity (RH 90 to 95%) at 22 ± 1°C, typical symptoms of brown rot developed on inoculated fruit, while control fruit remained symptomless. Isolates recovered from symptomatic fruit showed the same morphological and molecular characteristics as original isolates. To the best of our knowledge, this is the first report of M. polystroma in Serbia. Further studies are necessary to estimate the economic importance and geographic distribution of this organism in Serbia. References: (1) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (2) M. Hilber-Bodmer et al. Plant Dis. 96: 146, 2012. (3) G. C. M. van Leeuwen et al. Mycol. Res. 106: 444, 2002. (4) OEPP/EPPO Reporting Service. Retrieved from http://archives.eppo.int/EPPOReporting/2011/Rse-1106.pdf.

First Report of Brown Rot Caused by Monilinia fructicola on Stored Apple in Serbia.

Monilinia fructicola (G. Winter) Honey is a causal agent of brown rot of stone fruits, occasionally affecting pome fruits as well. The pathogen is commonly present in North and South America, Oceania, and Asia, but listed as a quarantine organism in Europe (4). After its first discovery in France in 2001, its occurrence has been reported in Germany, Hungary, Italy, Poland, Romania, Slovenia, Spain, Switzerland, Austria, and the Slovak Republic (1). In February 2011, during a survey for fungal postharvest pathogens in cold storage conditions, apple fruits (Malus domestica Borkh.) grown and stored in the Grocka Region, Serbia, were collected. All pathogens from symptomatic fruits were isolated on potato dextrose agar (PDA). One isolate from apple fruit cv. Golden Delicious with brown rot symptoms was identified as M. fructicola based on morphological and molecular characters. Colonies cultivated on PDA at 22°C in darkness were colorless, but later became grayish, developing mass of spores in concentric rings. Colony margins were even. Conidia were one-celled, limoniform, hyaline, measured 12.19 to 17.37 (mean 13.8) × 8.62 to 11.43 µm (mean 9.9), and were produced in branched monilioid chains (3). Morphological identification was confirmed by PCR (2) using genomic DNA extracted from the mycelium of pure culture, and an amplified product of 535 bp, specific for the species M. fructicola, was obtained. Sequence of the ribosomal (internal transcribed spacer) ITS1-5.8S-ITS2 region was obtained using primers ITS1 and ITS4 and deposited in GenBank (Accession No. JN176564). Control fruits were inoculated with sterile PDA plugs. After 3 days of incubation in plastic containers with high humidity at room temperature, typical symptoms of brown rot developed on inoculated fruits, while control fruits remained symptomless. The isolate recovered from symptomatic fruits showed the same morphological and molecular features of the original isolate. To our knowledge, this is the first report of M. fructicola in Serbia. Further studies are necessary for estimation of economic importance and geographic distribution of this quarantine organism in Serbia. References: (1) R. Baker et al. European Food Safety Authority. Online publication. www.efsa.europa.eu/efsajournal . EFSA J. 9(4):2119, 2011. (2) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (3) J. E. M. Mordue. CMI Descriptions of Pathogenic Fungi and Bacteria. No. 616, 1979. (4) OEPP/EPPO. EPPO A2 List of Pests Recommended for Regulation as Quarantine Pests. Online publication. Version 2010-09. Retrieved from http://www.eppo.org/QUARANTINE/listA2.htm , June 27, 2011.

First Report of Agrobacterium vitis as the Causal Agent of Grapevine Crown Gall in Serbia.

In November 2010, a serious outbreak of crown gall disease was observed on 3-year-old grapevine (Vitis vinifera L.) cv. Cabernet Sauvignon grafted onto Kober 5BB rootstock in two commercial vineyards located in the South Banat District in Serbia. Large, aerial tumors were visible above the grafting point on grapevine trunks, and in most cases, the tumors completely girdled the trunk. From the gall tissues, white, circular, and glistening bacterial colonies were isolated on yeast mannitol agar medium. Eight, nonfluorescent, gram-negative, and oxidase-positive strains were isolated from seven tumor samples and selected for further identification. PCR assays with A/C' (1) and VCF3/VCR3 (4) primers corresponding to the virD2 and virC genes yielded 224- and 414-bp fragments, respectively, confirming that the strains harbored the plasmid responsible for pathogenicity. The strains were differentiated to the species/biovar level with a multiplex PCR assay targeting 23S rRNA gene sequences (3) and were identified as Agrobacterium vitis. The 16S rDNA gene sequence from one isolate (GenBank Accession No. JN185718) showed 99% identity to the sequences of A. vitis previously deposited in NCBI GenBank database. The physiological and biochemical test results corresponded to the results of genetic analysis (2). The strains grew at 35°C and in nutrient broth supplemented with 2% NaCl. They were negative in 3-ketolactose, acid clearing on PDA supplemented with CaCO3, and ferric ammonium citrate tests; nonmotile at pH 7.0; pectolytic at pH 4.5; utilized citrate; produced acid from sucrose and alkali from tartarate. Pathogenicity was confirmed by inoculation of three plants per bacterial strain on grapevine cv. Cabernet Franc and on a local cultivar of tomato (Lycopersicon esculentum L.). The plants were inoculated on the stem by pricking one to three times through a drop of inoculum (108 CFU/ml) at three inoculation sites. Sterile distilled water was used as a negative control. Inoculated plants were maintained in a greenhouse at 24 ± 3°C. Typical tumors developed at the inoculation sites on tomatoes 3 weeks after inoculation and on grapevine 6 weeks after inoculation. No symptoms were observed on the control plants. Bacteria were reisolated from tumorigenic tissues and identified as pathogenic A. vitis by PCR. Crown gall disease was sporadically observed in vineyards in Serbia in previous years, but did not cause significant damage. Therefore, the causal agent was not studied in detail. To our knowledge, this is the first report of A. vitis determined as the causal agent of grapevine crown gall in Serbia. References: (1) J. H. Haas et al. Appl. Environ. Microbiol. 61:2879, 1995. (2) L. W. Moore et al. Page 17 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. N. W. Schaad et al., eds. The American Phytopathological Society, St. Paul, MN, 2001. (3) J. Pulawska et al. Syst. Appl. Microbiol. 29:470, 2006. (4) K. Suzaki et al. J. Gen. Plant Pathol. 70:342, 2004.

Prevalencia de consumo de drogas en estudiantes universitarios que cursan primer y cuarto año / Prevalence of tabacco, alcohol and marijuana consumption among university students

Background: University students are especially vulnerable towards substance abuse Aim: To describe and compare drug consumption in students of a Chilean university who attended first and fourth year of studies, according to gender and faculty. Material and Methods: A representative, stratified and proportional sample of305 students was randomly chosen by faculty, career, grade and gender during2009. The consumption of alcohol, tobacco and illicit drugs was evaluated using an anonymous survey. Results: Students from health care faculties had the lower prevalence of consumption of tobacco, alcohol, tobacco-alcohol and marijuana, during the first and fourth year. Education area and social sciences faculties had the highest prevalence of consumption. Fourth year students had higher rates of consumption than their first year counterparts. Females had significantly lower rates of alcohol and marijuana consumption. Conclusions: Alcohol, tobacco and marijuana consumption was higher among students from education and social sciences faculties and those attending the fourth year of studies.

[Prevalence of tabacco, alcohol and marijuana consumption among university students]. / Prevalencia de consumo de drogas en estudiantes universitarios que cursan primer y cuarto año.

BACKGROUND: University students are especially vulnerable towards substance abuse AIM: To describe and compare drug consumption in students of a Chilean university who attended first and fourth year of studies, according to gender and faculty. MATERIAL AND METHODS: A representative, stratified and proportional sample of 305 students was randomly chosen by faculty, career, grade and gender during 2009. The consumption of alcohol, tobacco and illicit drugs was evaluated using an anonymous survey. RESULTS: Students from health care faculties had the lower prevalence of consumption of tobacco, alcohol, tobacco-alcohol and marijuana, during the first and fourth year. Education area and social sciences faculties had the highest prevalence of consumption. Fourth year students had higher rates of consumption than their first year counterparts. Females had significantly lower rates of alcohol and marijuana consumption. CONCLUSIONS: Alcohol, tobacco and marijuana consumption was higher among students from education and social sciences faculties and those attending the fourth year of studies.

First Report of Occurrence of Benomyl Resistance in Botrytis cinerea Isolates on Raspberry in Serbia.

Botrytis fruit rot, caused by Botrytis cinerea, is one of the major diseases limiting production of raspberries (Rubus idaeus) in Serbia. Yield losses in commercial fields can exceed 50%, especially during periods of rainy, wet weather before harvest. Development of resistance to fungicides with site-specific modes of action is a serious problem in the control of B. cinerea worldwide. To insure the longest possible useful life of a fungicide, an early detection of shifts of sensitivity in pathogen population is crucial (1). The goal of this study was to evaluate sensitivity of B. cinerea isolates from commercial raspberry fields in Serbia to several fungicides that are frequently used: vinclozolin, benomyl, pyrimethanil, and fenhexamid. Initial isolation was done from sporulating berries during harvest. Single-spore isolates were identified based on colony and conidial morphology and by PCR amplification of an expected 0.7-kbp DNA fragment using B. cinerea-specific primer pair C729+/729- (3). Sensitivity of 130 isolates from six localities (20 to 30 isolates per locality) was determined on potato dextrose agar (PDA) amended with fungicides at discriminatory concentrations (1 and 10 mg/liter). Fungicides were suspended in sterile distilled water and added to autoclaved media that had cooled to 50°C. Inverted mycelial plugs (10-mm diameter), which had been cut from the edge of 4-day-old colonies on PDA, were placed on fungicide amended media and incubated for 48 h at 20°C. Treatments were replicated four times and the experiment repeated once. Strain SAS 56, which is sensitive to benzimidazoles and dicarboximides, and strain SAS 405, which is resistant to these fungicide classes, originating from German Collection of Microorganisms and Cell Cultures, were used as standards in the experiment. Isolates that did not grow at 1 mg/liter were designated as sensitive, those that grew at 10 mg/liter were considered highly resistant, and those that grew at 1 mg/liter but not at 10 mg/liter were classified as weakly resistant to all fungicides tested. Values of EC50 for all highly resistant strains were determined in radial growth experiments on PDA supplemented with a range of concentrations (5,000, 2,500, 1,000, and 500 mg/liter) of benomyl or thiophanate-methyl, according to the method described by Leroux and Gredt (2). All tested isolates were sensitive to vinclozolin, pyrimethanil, and fenhexamid. Nine of 130 isolates were highly resistant to benomyl with EC50 values between 1,056 and 1,523 mg/liter. The reference strain SAS 56 had an EC50 value of 0.17 mg/liter, compared to an EC50 value for SAS 405 strain of 1,548 mg/liter. All benomyl resistant isolates were also resistant to thiophanate-methyl and EC50 values ranged from 2,328 to 7,699 mg/liter. To our knowledge, this is the first report of benomyl resistance in isolates of B. cinerea on raspberry in Serbia. References: (1) H. Ishii. Jarq 40:205, 2006. (2) P. Leroux and M. Gredt. Page 1 in: Laboratoire de Phytopharmacie, INRA, Versailles, 1972. (3) S. Rigotti et al. FEMS Microbiol. Lett. 209:169, 2002.

Fentanyl analogs: structure-activity-relationship study.

Fentanyl is the prototype of the 4-anilidopiperidine class of synthetic opioid analgesics. This study was aimed to review the structure-activity-relationship (SAR) of fentanyl analogs substituted in the position 3, or 4 of the piperidine ring. Pharmacological results show that the groups in position 3 of the piperidine ring, which are larger than methyl, severely reduce the analgesic potency compared to fentanyl. It is likely that the steric factor alone (i.e. voluminosity of the group and cis/trans isomerism), rather than the polarity and/or chemical reactivity, plays a crucial role in the analgesic potency of this series. Although the duration of action, in general, does not depend on the stereochemistry, longer action of the most potent 3-alkyl fentanyl analogs such as cis-3-methyl- and cis-3-ethyl fentanyl, is more likely influenced by pharmacodynamic, rather than pharmacokinetic variables. Also, it is possible that the introduction of a functional group such as 3-carbomethoxy reduces the duration of action by altering pharmacokinetic properties. SAR findings obtained by evaluating the neurotoxic effects of fentanyl analogs substituted in the position 3 of the piperidine ring parallel the SAR findings on analgesia in regard to potency and duration of action. This might suggest that similar receptors are involved in producing both antinociceptive and neurotoxic effects of these drugs. It appears that both the potency and the duration of action in the series of fentanyl analogs substituted in position 4 of the piperidine ring is influenced only by the steric requirement and not by the chemical nature of the substituent.

First Report of Pear Decline Phytoplasmas on Pear in Serbia.

During August of 2004, pear (Pyrus communis L.) plants with typical symptoms of pear decline (PD) were observed in orchards in central Serbia. The affected plants showed premature reddening and upward rolling of leaves that often showed down-turned petioles. In some cases, premature defoliation was observed. Although a similar decline of pear was observed earlier, until now, the causal agent had not been identified. DNA was extracted with a chloroform/phenol procedure from fresh leaf midribs and branch phloem scrapes of four symptomatic and one asymptomatic pear plants separately. A nested polymerase chain reaction assay (PCR) was used for phytoplasma detection (first PCR round with P1/P7 (4) phytoplasma universal primer pair, followed by nested PCR with group 16SrX specific primers f01/r01) (3). With these primers, the expected products from phloem scrapes and midrib extracts of symptomatic plant samples were obtained. Restriction fragment length polymorphism (RFLP) analyses of the f01/r01 amplicon, with RsaI and SspI restriction enzymes, discriminating among 16SrX subgroup phytoplasmas, showed profiles corresponding to those of the apple proliferation phytoplasma group, 16SrX-C subgroup, "Candidatus Phytoplasma pyri" (2). A 1,155-bp sequence of 16S rDNA gene for one of the PA2f/r (1) amplicons obtained in nested PCR on P1/P7 products from one of the leaf midrib samples was deposited in GenBank (Accession No. AY949984); both strands of the fragment were sequenced with the Big Dye Terminator reaction kit (Applied Biosystems, Foster City, CA). The sequences were analyzed with the Chromas 1.55 DNA sequencing software (Technelysium, Queensland, Australia) and aligned with BLAST software ( http://www.ncbi.nlm.nih.gov ). The blast search showed 100% homology of this sequence with that of PD strain Y16392, confirming the identity with PD of the phytoplasma detected. To our knowledge, this is the first report of pear decline phytoplasmas in Serbia. References: (1) M. Heinrich et al. Plant Mol. Biol. Rep. 19:169, 2001. (2) IRPCM Phytoplasma/Spiroplasma Working Team-Phytoplasma Taxonomy Group. Int. J. Syst. Evol. Microbiol. 54:1243, 2004. (3) K.-H. Lorenz et al. Phytopathology 85:771, 1995. (4) Schneider et al. Pages 369-380 in: Molecular and Diagnostic Procedures in Mycoplasmology. Vol I. S. Razin and J. G. Tully, eds. The American Phytopathological Society, 1995.

First Report of an Elm Yellows Subgroup 16SrV-C Phytoplasma Infecting Grapevine in Serbia.

During a 2002 survey in Serbia, samples of grapevine (Vitis vinifera) were collected from plants showing typical phytoplasma-like symptoms: leaf roll, leaf redness, vein chlorosis and necrosis, and absence of lignification. The material was collected from one viticultural region (Zupa Aleksandrovac), where the disease was recorded in 2000 and showed an increasing percentage of symptomatic plants every year. Total nucleic acid was extracted separately from leaf midveins and stem bark collected from 10 symptomatic and 2 asymptomatic plants. Phytoplasma infection was detected using polymerase chain reaction (PCR) assays with universal primer pair P1/P7 for the amplification of phytoplasma 16S rRNA gene, and primer pair FD9f2/FD9r followed by FD9f3/FD9r2 in nested PCR for specific amplification of the FD9 nonribosomal DNA fragment of the EY-group (1). Phytoplasmas were detected in 9 of 10 midvein extracts from symptomatic grapevines (three of cv. Plovdina, two of cv. Smederevka, and four of cv. Gamé). Also, 6 of 10 bark preparations representing stem collections from the same plants were positive (two samples of cv. Plovdina, both samples of cv. Smederevka, and two samples of cv. Gamé). Both collections of midveins and bark tissues from asymptomatic plants were negative. Fragments amplified with universal P1/P7 primers (16S-23S rDNA) were analyzed by restriction fragment length polymorphism with TruI and TaqI restriction enzymes. The phytoplasmas produced identical restriction profiles to those of 16SrV Elm Yellows group and 16SrV-C Flavescence doreé subgroup (2). To our knowledge, this is the first report of phytoplasma infecting grapevines in Serbia, and the first survey in progress to verify the presence of Scaphoideus titanus to determine if this grapevine yellows could be defined as Flavescence dorée. References: (1) E. Angelini et al. Vitis 40:79, 2001. (2) M. Martini et al. Mol. Cell. Probes 16:197, 2002.

Detection of motion in hybrid PET/SPECT imaging based on the correlation of partial sinograms.

This paper describes a motion detection method specific to hybrid positron emission tomography/single photon emission computed tomography systems. The method relies on temporal fractionation of the acquisition into three data sets followed by an algorithm based on the cross correlation (CC) of partial sinograms from successive sets at different rotations of the camera. Spatial inconsistencies due to motion are detected by decreases in the CC between two sets. This permits to separate data into premotion and postmotion sets of consistent data that are reconstructed independently then registered and summed. Rigid motions greater than 1-cm translation or 10 degrees rotation were detected with this method from experimental data obtained by manually moving phantoms made of radioactive spheres as well as from a patient lung study corrupted by artificial motion. The different motion studies showed that the image contrast does not seem to be a limiting factor and that the motion is best detected when the gantry is parallel to the direction of motion. The registration and fusion of the reconstructed premotion and postmotion sets lead in all cases to a reduction of the motion artifacts and an increase in signal-to-noise ratio.

Functional, dynamic, and anatomic MR urography: feasibility and preliminary findings.

RATIONALE AND OBJECTIVES: The authors assessed the feasibility of using magnetic resonance (MR) urography to acquire functional, dynamic, and anatomic information in human subjects with normal and hydronephrotic kidneys. MATERIALS AND METHODS: In subjects known to have or suspected of having hydronephrosis, split renal filtration fractions were measured with a customized magnetization-prepared, inversion-prepared gradient-recalled echo sequence to determine the T1 of flowing blood in the inferior vena cava and aorta before and after contrast medium administration and in the renal veins and arteries after contrast medium administration. Multiple timed sets of coronal fast spoiled gradient-echo 70 degrees flip-angle images were acquired before and after contrast medium administration to derive MR renograms from changes in the signal intensity of the cortex and medulla. Precontrast T2-weighted images were obtained with a three-dimensional fast spoiled gradient-echo maximum intensity projection pulse sequence, and postcontrast T1 maximum intensity projection images were also obtained to depict the renal anatomy. RESULTS: Split filtration fraction differentiated normal from hydronephrotic kidneys. MR renograms depicted vascular, tubular, and ductal phases and differentiated between normal and hydronephrotic kidneys (P < .05, n = 20). Contrast medium dose correlated with the peak of the cortical signal intensity curves on the renogram (r = 0.7, P < .0005; n = 20). The sensitivities for the visual determination of hydronephrosis and unilateral delayed excretion of contrast material were both 100%, and the specificities were 64% and 85%, respectively. CONCLUSION: The preliminary findings show promise for the use of MR urography in the comprehensive assessment of renal function, dynamics, and anatomy.

Antinociceptive activity of the novel fentanyl analogue iso-carfentanil in rats.

A large number of fentanyl analogues have been synthesized so far, both to establish the structure-activity-relationship (SAR) and to find novel, clinically useful antinociceptive drugs. In this study, the newly synthesized fentanyl analogue 3-carbomethoxy fentanyl (iso-carfentanil) was compared to fentanyl for its antinociceptive activity (tail-immersion test) in rats. It was revealed that the introduction of a 3-carbomethoxy group in the piperidine ring of fentanyl skeleton reduced the potency and shortened the duration of action of the parent compound, i.e., fentanyl. The antinociceptive potency of 3-carbomethoxy fentanyl is influenced mainly by the steric factor (voluminosity of the carbomethoxy group and the cis/trans isomerism), while the chemical nature of the group is probably irrelevant. This is in agreement with SAR studies of other 3-substituted fentanyl analogues. In contrast to potency, the duration of action is not affected by cis/trans isomerism. It is assumed that the time course of action of 3-carbomethoxy fentanyl is influenced by the nature of the carbomethoxy group. Since the potency and the duration of action of this novel antinociceptive compound are interesting from the aspect of SAR studies and have potential promise for clinical application, 3-carbomethoxy fentanyl deserves to be extensively evaluated.