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Objectives: The diagnosis of bloodstream infections (BSIs) in immunocompromised patients, such as patients with cancer, is challenging. Although blood culture (BC) is considered the standard diagnostic tool for BSIs, it takes several days to yield results and has low sensitivity in these patients. Here, we tested a novel method for diagnosing BSIs in a large cohort of immunodepressed patients., Methods: Real-time PCR (LightCycler SeptiFast Test M(GRADE), Roche Diagnostics) was compared with BC for its ability to detect bacteria and fungi in blood samples from 100 immunocompromised patients (98 with cancer) in whom sepsis was suspected., Results: In concordant samples (79.2% of total cases), real-time PCR identified the presence or absence of microbes significantly faster than BC (p=3.7x10(-49), t-test). Furthermore, in 6 cases, SeptiFast distinguished contamination of BCs by coagulase-negative staphylococci. SeptiFast, however, failed to detect 5 cases of clinically relevant BSI that tested positive by BC., Conclusions: SeptiFast rapidly diagnosed BSIs in our cohort of immunosuppressed patients. The results of this study suggest that SeptiFast can be used in conjunction with, but cannot replace, BC to better identify the etiology of fever in immunocompromised patients.

Rare earth elements (REEs) enriched fertilizers have been commonly used in China since the 1980s, thus inducing a growing concern about their environmental impact in agriculture. In this work, the effect of some light REEs nitrate mixture and La(3+) nitrate on seed germination, seedling growth and antioxidant metabolism in Triticum durum was investigated with the aim of clarifying the potential benefits or damages of REEs on plants. Seed pre-soaking for 8 h with La(3+) and REEs nitrate inhibited seed germination at low concentrations (0.01 mM and 0.1 mM), while pre-soaking for 2 and 4 h already inhibited seed germination when higher concentrations (1 mM and 10 mM) of La(3+) and REEs nitrate were used. La(3+) and REEs nitrate treatment also affected seedling growth. Root growth was enhanced and inhibited at low and high concentrations, respectively. Shoot growth was inhibited by La(3+) and REEs nitrate at all tested concentrations after 12 d of treatments. Enzymatic and non enzymatic antioxidants were differently affected by La(3+) and REEs nitrate and their behaviour changed also depending on the plant organ. In roots La(3+) and REEs nitrate treatments induced an increase in ascorbate (ASC) and glutathione (GSH) contents. In shoots only La(3+) nitrate induced an increase in the ASC content whereas GSH decreased following both La(3+) and REEs nitrate treatments. An increase in ASC peroxidase activity was observed in shoots and roots, while catalase did not change in roots and slightly decreased in shoots. The possible role of the increase in some antioxidants as indicators of stress caused by lanthanide treatments is discussed.

The expression of exogenous antibodies in plant is an effective strategy to confer protection against viral infection or to produce molecules with pharmaceutical interest. However, the acceptance of the transgenic technology to obtain self-protecting plants depends on the assessment of their substantial equivalence compared to non-modified crops with an established history of safe use. In fact, the possibility exists that the introduction of transgenes in plants may alter expression of endogenous genes and/or normal production of metabolites. In this study, we investigated whether the expression in plant of recombinant antibodies directed against viral proteins may influence the host leaf proteome. Two transgenic plant models, generated by Agrobacterium tumefaciens-mediated transformation, were analyzed for this purpose, namely, Lycopersicon esculentum cv. MicroTom and Nicotiana benthamiana, expressing recombinant antibodies against cucumber mosaic virus and tomato spotted wilt virus, respectively. To obtain a significant representation of plant proteomes, optimized extraction procedures have been devised for each plant species. The proteome repertoire of antibody-expressing and control plants was compared by 2-DE associated to DIGE technology. Among the 2000 spots detected within the gels, about 10 resulted differentially expressed in each transgenic model and were identified by MALDI-TOF PMF and muLC-ESI-IT-MS/MS procedures. Protein variations were restricted to a limited number of defined differences with an average ratio below 2.4. Most of the differentially expressed proteins were related to photosynthesis or defense function. The overall results suggest that the expression of recombinant antibodies in both systems does not significantly alter the leaf proteomic profile, contributing to assess the biosafety of resistant plants expressing antiviral antibodies.

Objective: To evaluate self and parent reports on quality of life (QoL) and psychological adjustment of youths with type 1 diabetes, in comparison to a general paediatric population, and identify relationships between disease duration, metabolic control and psychological parameters., Research Design and Methods: Participants included 70 youths with type 1 diabetes and their parents. They were compared with 70 non-diabetic subjects. Data were analyzed in the whole group and in subgroups aged 6-10, 11-13 and 14-18 yr. All cases performed pediatric QoL, Child Behaviour Checklist, filled in by parents, and Youth Self-Report, filled in by youths. Data were compared with haemoglobin A1c (HbA1c) values and disease duration., Results: Self-reports showed a psychological adjustment of youths with type 1 diabetes similar to that of controls. Parent reports showed that parents of children with type 1 diabetes were more worried than those of controls (p < 0.01). Adolescents showed a worse QoL and more frequent psychological disturbances. In this group, for youth and parent reports, HbA1c levels correlated positively with psychological problems (p < 0.05) and negatively with QoL (p < 0.05). Only for parent reports, in the whole group and in subgroups aged 6-10 and 11-13 yr, disease duration correlated positively with psychological adjustment (p < 0.05)., Conclusions: Before adolescence, youths with type 1 diabetes showed only slight problems in psychological adjustment and QoL, with an association with disease duration reported by parents. In adolescence, both youths and their parents reported more emotional and behavioural problems, independent of disease duration. Better metabolic control and psychological well-being seemed directly related.

Background & Aims: There is scarce information about immune function and parenteral fish oil (FO). The influence of a new parenteral lipid emulsion (LE) containing fish oil (SMOF) was experimentally evaluated on neutrophils' chemotaxis and macrophages' phagocytosis., Methods: Adult male Lewis rats (n=40) were randomized into five groups; one non-surgical control and four to receive parenteral LE or saline infusion through jugular vein catheterization: SMOF (mixture of 30% medium-chain triglycerides, 30% soybean, 25% olive and 15% fish oils); MCT/LCT (physical mixture of 50% medium-chain triglycerides and 50% soybean oil); MCT/LCT/FO (80% MCT/LCT supplemented with 20% FO) and SS (saline). In the 5th experimental day and after intravenous colloidal carbon injection, blood and tissue (liver, lung and spleen) samples were collected and immunological analyses were performed., Results: LE didn't influence neutrophil chemotaxis. SMOF didn't influence phagocytosis (p>0.05) while MCT/LCT and MCT/LCT/FO LE increased the number of liver and lung resident macrophages that had engaged in phagocytosis compared with CO-NS and SS (p<0.05). Only MCT/LCT/FO increased the number of spleen resident macrophages that had engaged in phagocytosis (p<0.05)., Conclusions: LE, independently of composition, had no influence on neutrophils' chemotaxis, but showed different effect on phagocytosis by macrophages. SMOF LE had neutral effect while fish oil LE enriched with MCT/LCT LE increased resident-macrophages' phagocytosis.

Both medical doctors and humanitarian operators engaged in health relief or development missions abroad, are called to respect the general principles of international law, that is to say, customary law that is legally compulsory for the International Community and rules deriving from Treaties and International Conventions. Humanitarian operators have to observe also the rules and regulations of the hosting country. They have to respect all rules applying to their humanitarian action and they have to take responsibility towards beneficiaries and donors alike.

A disaster both natural and man made is managed in time according to separate phases that respond to specific requirements and needs. In the late and post emergency phases problems of public health are priority and in particular the identification and containment of epidemic clusters. Epidemiological surveillance is the main instrument used and in this paper it is analysed both in theory and in its application in the Italian Red Cross mission in Sri Lanka devastated by the 2004 Tsunami.

Facing the numerous humanitarian emergencies that upset the international balances, the International Organisations (IO) who intervene are: the Agencies of the United Nations (UN), the International Federation of the Red Cross (IFRC) and NGOs. In the sector of international health cooperation, the World Health Organization (WHO) has elaborated a document, the International Health Regulations (IHR), that regulates the health workers' activity during a health emergency. In this paper different International Organizations employed in international humanitarian crisis are described.

Recent studies show how in the 2004-2005 period there has been an increase in natural disasters of 18% worldwide. According to a renowned author planning for disaster response is as valid as the starting hypothesis. The study of an inductive mental process in disaster response planning is the key to avoiding the invention and re-invention of the wheel for each emergency. Research in this field however is hampered by different factors one of which is data collection that during disaster response requires specific training. Standardization of data collection models with limitation of the number of variables is required as is taking into account problems related to people migration and subsequent sampling problems and retrospective analysis. Moreover poor attention to the training of the volunteers employed on the field is an issue to be considered.

Risk perception is due to a subjective evaluation of reality and depends on a number of factors such as: individual coherence and flexibility, the set of social values and rules that contributed to the definition of the individuals' interior world - this does not always make proper risk perception possible. Even though the way common people set-up their own judgment is based on their own interpretation tracks, surely different from expert criteria, we cannot label their approach as irrational. Over time, through an evolutionary process, people have decided what is safe and what is dangerous; this personal awareness was not based on the science but on experiences, images, feelings and emotions. Most of the time, neither social nor economical conditions can prevent people to feel an epidemic risk in the same way as it was felt during the plague of 1350. Most of the time, after a disaster, victims experience a regression so, for example, even if it is known that corpses do not always cause infections, after the tsunami disaster, most of the bodies were quickly cremated by scared survivors; this made the identification and burial of the victims impossible.

The benzene, toluene, ethylbenzene and xylenes system (BTEX) in clean water is studied to verify the performance of capillary extraction as an extraction-preconcentration technique well hyphenated with GC. The approach uses pieces of coated capillaries usually 5-30 cm long, trimmed from customary high-resolution GC columns but carrying glass press-fits at their ends. The preparation of these 'capillary extractors' is explained, and their performance is discussed providing guidelines for use. Injection by capillary extraction is such that (i) band broadening in time is null, and (ii) band broadening in space cannot be higher than the extractor length. Speed, cleanliness and operative simplicity of the capillary extraction approach are remarkable, pros and cons are complementary to those of solid phase microextraction (SPME) or stir bar sorptive extraction (SBSE). Capillary extraction-capillary GC analysis of aqueous BTEX samples, in a clean water matrix, allows low part-per-billion detection limits, and does not require heated injectors or cryofocusing devices.

Capillary extractors are proposed as samplers-preconcentrators to overcome losses of volatile organic compounds found using "classical" solid-phase microextraction fiber-holder samplers. A set of equal-size extractors was used to extract in-tube an aqueous solution of benzene, toluene, ethylbenzene and xylenes (BTEX) at 146 ppb. After storage for 6-30 days at 0-4 degrees C (or -15 degrees C) GC analyses were carried out to study BTEX recovery. Results demonstrated that sample preservation was very good: recovery was higher than 98% after 6 days, and more than 95% after 30 days. Capillary extractors, due to their high performance in preserving sample integrity, represent a real breakthrough for on-site sampling of volatile compounds by solid-phase pre-concentration techniques.

The suitability of "capillary extractors" is demonstrated for the "negligible depletion" extraction of benzene, toluene, ethylbenzene and xylenes in a clean-water matrix. Extraction set-up and major extractor parameters (length, internal diameter, and film thickness) are chosen to allow rugged analysis by GC with flame ionization detection. With the selected negligible extraction conditions, the efficiency for every consecutive extraction is about 2-3% of the dissolved amount.

Partition coefficients of benzene, toluene, ethylbenzene and xylenes (BTEX), between crosslinked polydimethylsiloxane and water, were determined at room temperature by capillary extraction (a form of in-tube solid-phase microextraction, SPME) coupled to open tubular gas chromatography (in-tube SPME-high-resolution GC). A series of 7-9 repetitive extractions, performed on a 1-ml volume of diluted aqueous BTEX sample by the double-syringe squeeze method, gave exponential regression curves which fit very well with those predicted by partition theory. From the equations of the curves of relative FID response vs. extraction number, experimental Kd were easily calculated and the results compared with literature values. The whole measurement requires about 1 h from the start of the experiment to the final calculation of all BTEX partition coefficients. In-tube SPME resulted in a fast, clean, efficient, and cheaper alternative than the classic 1-cm, externally coated, SPME fiber-holder technique.

"Breakthrough" during static coating could be avoided using a new coating strategy: while filling the capillary support with coating solution, a short mercury plug was placed at the middle of the support length. Mercury was frozen by a cold source and vacuum was applied to both open ends. In this way static coating became symmetric and so evaporation rate was doubled, on a time base, and more than doubled on a coated length base. Coating time at room temperature varied from approximately 20 min for 3-m columns to approximately 3 h for 15-m columns.

The construction of a high-efficiency but cheap injector for volatile and very volatile compounds is shown. The device focuses the compounds in a fused-silica (FS) transfer capillary with the aid of liquid nitrogen. A 6.2 mm O.D. glass tube liner (ca. 25 cm x 1.5 mm I.D.) is inserted in the heated (-200 degrees C) injector of the gas chromatograph in place of the standard glass liner, and extends further externally through a liquid nitrogen container made with styrofoam-like material. Inside this glass tube, the FS transfer line passing through the oven door is connected like a pre-column to the analytical high-resolution GC column. It can move fast between the heated and the cooled zone (<-->, deltaL = 13 cm), and when this movement starts, crvofocused analytes are injected "at once" resulting in symmetrical and sharp injection bands with "zero" carryover. The performance of this device is demonstrated by its application to in-tube solid-phase microextraction and to spice volatiles analysis.

We describe a protocol for flow cytometry analysis of cell cycle in plants using indirect immunolabelling staining and Vicia faba, Pisum sativum and Zea mays root tip cells as model systems. The protocol is based on simultaneous analysis of two fluorescent signals. The first, obtained after staining with propidium iodide, is used to quantify nuclear DNA content. The second, obtained after indirect immunofluorescent staining of bromodeoxyuridine (BrdUrd), is used to quantify the amount of BrdUrd incorporated into nuclear DNA. In an attempt to standardize the procedure, the effects of various conditions for partial DNA denaturation using HCl, as well as of BrdUrd concentration and incorporation time on flow cytometry DNA/BrdUrd content analysis have been studied. Maximum BrdUrd-linked fluorescence was observed after a 30 min pulse with 10 microM BrdUrd and after DNA denaturation with 1.5 N HCl (final concentration) for 30 min at 25 degrees C. Under these conditions, DNA content histograms with relatively small coefficient of variation (< 4%, full peak) could be obtained. To avoid non-specific staining of cytoplasm and cell walls, the protocol involves the use of nuclei isolated from formaldehyde-fixed tissues. Fixed isolated nuclei are stable and may be stored in hexylene glycol 0.75 M at 4 degrees C for prolonged periods prior to actual staining and analysis.

The authors base their remark on a few cases of awareness which have been reported in their hospital and on the history a patient who experienced intraoperative awareness during cesarean section. Attention is focused on the ethical, social and medico-legal aspects of this phenomenon. Awareness may severely affect the patient's psychic equilibrium with the consequent onset of severe neuroses leading to the possibility of legal action and claims for compensation. Clinical experience has shown that the use of halogenate anesthetics, even in low concentrations, may help to avoid this phenomenon. Correct relations between the anesthetist and patient before and after surgery may also, even in the event of awareness, reduce the incidence and severity of consequent psychological sequelae.

It is mandatory, the Authors say, to obtain a previous and definite consent before the patient undergoes the operation. Moreover the patient shall be well acquainted about the risk connected to the operation before he gives his consent. On the contrary, as already confirmed by the Italian Supreme Court of Justice, the consent shall not be valid and the Doctors could be found guilty of "psychic violence". Finally the Authors, to evaluate the probable risks liked to the operation, use scales as ASA and prognostic indexes as SAPS to avoid appraisage too subjective.

The measurement of recirculation during two-needle hemodialysis has been shown to provide valuable information concerning distal hemoaccess stenosis and the loss of effective dialysis. Reference sources have suggested that levels of recirculation above 20% call for further investigation. We proposed a technique for routine recirculation measurement and studied 20 stable hemodialysis patients using this method. We found that all patients had recirculations of less than 10%. We also studied 2 patients with angiographically proven stenosis and found 1 patient with a recirculation of less than 20%. Routine recirculation testing provides valuable information about hemoaccess integrity and using our techniques, recirculation greater than 10% is an indication for further study.