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La distribución de las especies de sínfitos y aculeados ha sido investigada por medio de seis trampas Malaise instaladas en dos series de vegetación adyacentes y sucesivas, bosque mixto de pinos y hayedo, en la reserva forestal de Arikutza (Navarra). La comparación entre series de vegetación y localización de las trampas en sínfitos, avispas, abejas y hormigas sugiere que la distribución de las especies puede estar relacionada con la distribución espacial de los distintos recursos de alimentación, reproducción, nidificación o hibernación. Los claros, el cauce del río y la madera muerta proveen variedad de fuentes de diversidad.

In line with the current interest in animal welfare in animal husbandry, this study of two groups of 24 sheep compares milking performances at a low vacuum level (28 kPa) and at a standard level (42 kPa) that is normally used for sheep. The two working vacuum levels were tested with three different combinations of pulsator rate and pulsator ratio. Each of the experiments lasted 21 days, of which 11 were for adaptation and 10 were for collecting data. The milking machine used was a low line with 24 stalls, 12 milking units controlled by two operators, a milk line 74 mm in diameter, and a cluster with a total mass of 490 g. The measurements performed were the dynamics of the vacuum level in the short milk tube and the milk line during milking, and the time duration for each individual milking. There were no significant differences in fluctuations in vacuum in either the short tube or the milk line at the two vacuum levels used. By contrast, there was a significant increase in vacuum fluctuations in the short milk tube when the pulsator ratio changed from 50% to 60%. The milk flow time for each sheep increased on average by 17% with low vacuum, while the throughput of the system was reduced by some 4%, from 333 to 321 sheep/h. [ABSTRACT FROM AUTHOR]

Bacterial resistance to antimicrobials is considered a major issue worldwide. This condition may account for treatment failure of urinary tract infections, which are among the most common infections both in community and healthcare settings. Therapy against uropathogens is generally administered empirically, possibly leading to unsuccessful therapy, recurrence and development of antibiotic resistance. The reduction in analytical time to obtain antimicrobial susceptibility test (AST) results could play a key role in reducing the cost of healthcare, providing information about antibiotic efficacy and thus preventing from either exploiting new and expensive antibiotics unnecessarily or using obsolete and ineffective ones. A more rational choice among treatment options would hence lead to more effective treatment and faster resolution. In this paper we evaluated the performance of a new Point Of Care Test (POCT) for the rapid prediction of antimicrobial susceptibility in urine samples performed without the need of a laboratory or specialized technicians. 349 patients were enrolled in two open-label, monocentric, non-interventional clinical trials in partnership with an Emergency Medicine ward and the Day Hospital of two large healthcare facilities in Rome. Antibiogram was carried out on 97 patients. Results from analysis of urine samples with the POCT were compared with those from routine AST performed on culture-positive samples, displaying high accuracy (>90%) for all tested antimicrobial drugs and yielding reliable results in less than 12 hours from urine collection thus reducing analytical and management costs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Arienzo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The aim of this study was to evaluate the performance of "Micro Biological Survey - MBS Test" in the enumeration of bacterial load in cow raw milk. The MBS test is based on a colorimetric method recently developed and patented by "Roma Tre" University, Italy. The evaluation of the performance of the MBS method was carried out by comparison with plate count at 30°C (gold standard) and flow cytometry. Thirteen independent set of experiments were performed analyzing a total of 104 samples of cow raw milk with the selected methods. Results obtained using the MBS method are comparable with those obtained with the plate count method at 30°C (CFU/mL) and flow cytometry technology; in particular, the results obtained with the MBS method are very close to plate count's at 30°C. On the other hand, there are statistically significant differences between these two methods' and flow cytometry technology's results that could be due to the different experimental conditions., Competing Interests: Conflict of interests: The authors declare no potential conflict of interests., (©Copyright: the Author(s).)

The market of ready-to-eat leafy green salads is experiencing a noticeable growth in Europe. Since they are intended to be consumed without additional treatments, these ready-to-eat products are associated with a high microbiological risk. The aim of this work was to evaluate the microbiological quality and safety of ready-to-eat leafy green salads sold in widespread supermarket chains in Lazio, Italy, on the packaging date during shelf-life and during home-refrigeration. The study also aimed to determine the differences between low-, medium-, and high-cost products. Salmonella spp. and L. monocytogenes were chosen as safety indicators as specified by European regulations while total aerobic mesophilic bacteria and Escherichia coli were chosen as quality indicators as suggested by national guidelines. Analyses were performed following the ISO standards and in parallel for the evaluation of total aerobic mesophilic bacteria, with an alternative colorimetric system, the Micro Biological Survey method, in order to propose a simple, affordable and accurate alternative for testing the microbiological quality of products, especially suitable for small and medium enterprises and on-site analyses. The study revealed high, unsatisfactory, total bacterial loads in all analyzed samples on the packaging date and expiry date and a very high prevalence of Salmonella spp. (67%) regardless of the selected varieties and cost categories; L. monocytogenes was not recovered aligning with the results obtained in other studies.

The consumption of dairy products and the dairy industry are one of the main global agri-food sectors for its size, economic importance, and level of technology. Microbiological quality of pasteurized milk or other milk products is dependent on microbiological quality of raw milk. A variety of microbiological count methods is available for monitoring the hygienic quality of raw milk. Among them, the pour plate method is the official essay for counting the number of colony-forming units in milk samples according to International Organization for Standardization (ISO) No. 4833-1:2013. The aim of the present study is the validation of the Micro Biological Survey (MBS) method, against the reference plate-count method, for the assessment of the microbiological quality of raw milk. This comparative study, performed in collaboration with the Istituto Zooprofilattico Sperimentale del Lazio e della Toscana M. Aleandri (IZSLT), demonstrates the accuracy of this alternative method for the determination of total viable bacterial count in cow's raw milk. The results obtained with the MBS method highlight its potential as a valid tool for reliable microbiological analysis in dairy industries.

Urinary tract infections (UTIs) are among the most common infections in all age groups. Fast and accurate diagnosis is essential to ensure a timely and effective therapy. Alongside with reference culture-based methods, several point-of-care tests (POCTs) for early detection of UTIs have been developed, but they have not been significantly implemented in current clinical practice. The Micro Biological Survey (MBS) POCT is a simple test developed by MBS Diagnostics Ltd. (London, UK) for the detection and management of UTIs. The present study has been undertaken to investigate the potentials and limits of the MBS POCT. A total of 349 patients were enrolled in two open-label, monocentric, non-interventional clinical trials in collaboration with an Emergency Medicine department and the outpatient clinic of two hospitals in Rome. Results of urine analysis using the MBS POCT were compared with those of the routine culture-based tests for UTI diagnosis performed by the hospital laboratory. The MBS POCT provided fast results revealing high bacterial count UTIs (≥ 10 5  CFU/ml) with 97% accuracy, 92% sensitivity, 100% specificity, 99% PPV, and 96% NPV within a 5-h analytical time threshold.

We focused on an integrated view of genomic changes in Colorectal cancer (CRC) and distant normal colon tissue (NTC) to test the effectiveness of expression profiling on identification of molecular targets. We performed transcriptome on 16 primary coupled CRC and NTC tissues. We identified pathways and networks related to pathophysiology of CRC and selected potential therapeutic targets. CRC cells have multiple ways to reprogram its transcriptome: a functional enrichment analysis in 285 genes, 25% mutated, showed that they control the major cellular processes known to promote tumorigenesis. Among the genes showing alternative splicing, cell cycle related genes were upregulated (CCND1, CDC25B, MCM2, MCM3), while genes involved in fatty acid metabolism (ACAAA2, ACADS, ACAT1, ACOX, CPT1A, HMGCS2) were downregulated. Overall 148 genes showed differential splicing identifying 17 new isoforms. Most of them are involved in the pathogenesis of CRC, although the functions of these variants remain unknown. We identified 2 in-frame fusion events, KRT19-KRT18 and EEF1A1-HSP90AB1, encoding for chemical proteins in two CRC patients. We draw a functional interactome map involving integrated multiple genomic features in CRC. Finally, we underline that two functional cell programs are prevalently deregulated and absolutely crucial to determinate and sustain CRC phenotype.

The JC polyomavirus (JCV) has been repeatedly but discordantly detected in healthy colonic mucosa, adenomatous polyps, and colorectal cancer (CRC), and proposed to contribute to oncogenesis. The controversies may derive from differences in JCV targets, patient's cohorts, and methods. Studies of simultaneous detection, quantification, and characterization of JCV presence/expression in paired samples of normal/altered tissues of the same patient are lacking. Therefore, we simultaneously quantified JCV presence (DNA) and expression (mRNA and protein) of T-antigen ( T-Ag ), Viral Protein 1 ( Vp1 ), and miR-J1-5p in paired normal/altered tissues of CRC or polyps, and from controls. JCV signatures were found in most samples. They increased in patients, but were higher in normal mucosa than in corresponding polyp or CRC lesions. JCV non-coding control region (NCCR) DNA rearrangements increased in CRC patients, also in normal mucosa, thus before the onset of the lesion. A new ∆98bp NCCR DNA rearrangement was detected. T-Ag levels were higher in normal mucosa than in adenoma and adenocarcinoma lesions, but decreased to levels of controls in established CRC lesions. In CRC, miR-J1-5p expression decreased with CRC progression. Vp1 expression was not detected. The data indicate a JCV link with the disease, but possible JCV contributes to oncogenesis should occur at pre-polyp stages.

Two common polymorphisms in the MTHFR gene, C677T and A1298C, are associated with reduced enzyme activity and may be associated with breast cancer susceptibility. We performed a case-control study to investigate the association between the two SNPs in the MTHFR gene and risk of breast cancer. In total, 58 breast cancer patients and 58 unaffected controls were enrolled in the study. Polymerase chain reaction/restriction fragment length polymorphism technique (PCR-RFLP) was conducted to determine the genotypes. No significant differences were found in the genotypes of the two polymorphisms of the MTHFR gene between cases and controls. The OR and 95% CI for the 677CC, 677CT and 677TT genotypes were 1.00, 0.95 (0.39-2.31) and 0.87 (0.27-2.80), respectively; those of the 1298AA, 1298AC and 1298CC genotypes were 1.00, 0.59 (0.26-1.36) and 0.78 (1.32-4.66) respectively. Furthermore, it has been shown in patients with breast cancer a risk of presenting with an aggressive biophenotype about twice or three times higher in the presence of the C677T and A1298C polymorphisms, respectively. Finally, the A1298Cpolymorphism is significantly associated with increased recurrence risk of lymph node-positive breast cancer. Our study has not shown a significant association between MTHFR gene polymorphisms and breast cancer risk. However, it highlighted the key-role played by the presence of mutant alleles for both polymorphisms in increasing the risk of developing more aggressive phenotypes; moreover, specifically in A1298C, it might also lead to a higher risk of developing lymph node metastasis., Competing Interests: Competing Interests: The authors have declared that no competing interest exists.

Over the last decades, traditional olive production has been converted to intensive and super-intensive cultivation systems, characterized by high plant density and irrigation. Although this conversion improves product quality and quantity, it requires a larger amount of energy input. The new contributions in this paper are, first, an analysis of the energy and environmental performance of two commercial-scale high peak-power hybrid photovoltaic irrigation systems (HPVIS) installed at intensive and super-intensive Mediterranean olive orchards; second, an analysis of PV hybrid solutions, comparing PV hybridization with the electric power grid and with diesel generators; and finally, a comparison of the environmental benefits of HPVIS with conventional power sources. Energy and environmental performances were assessed through energy and carbon payback times (EPBT and CPBT). The results show EPBT of 1.98 and 4.58 years and CPBT of 1.86 and 9.16 years for HPVIS in Morocco and Portugal, respectively. Moreover, the HPVIS were able to achieve low emission rates, corresponding to 48 and 103 g CO 2 e per kWh generated. The EPBT and CPBT obtained in this study were directly linked with the irrigation schedules of the olive orchards; therefore, weather conditions and irrigation management may modify the energy and environmental performances of HPVIS. The consumption of grid electricity and diesel fuel, before and after the implementation of HPVIS, was also analyzed. The results obtained show fossil energy savings of 67% for the Moroccan farm and 41% for the Portuguese installation. These savings suggest that the energy produced by HPVIS in olive orchards will avoid the emissions of a large amount of greenhouse gas and the exploitation of natural resources associated with fossil fuel production., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

Access to safe water is stated within human rights as essential for life, as water can be a source of severe enteric infections threatening human health, in particular children from Developing Countries. Along with reference methods, need is pressing for alternative methods to flank reference ones to improve water safety on-site monitoring and in the absence of scientific facilities or even electricity supply. The Micro Biological Survey (MBS) method has already been successfully applied to water safety assessment in Developing Countries. A total of 18 water samples were collected from different sources (rivers, dug wells, tap water) within the Rukwa Region, Tanzania, and underwent analysis for Total Coliforms following the MBS method. Globally, rivers showed more frequently contamination, followed by dug wells, tap water and tanks. Results demonstrate the need for continuous monitoring of water sources, even in difficult frameworks lacking electric supply, to help improve control over water quality, possibly using alternative methods to simplify existing protocols., Competing Interests: Conflict of interest: the authors declare no potential conflict of interest.

The clinical and genetic heterogeneity of Triple Negative Breast Cancer (TNBC) and the lack of unambiguous molecular targets contribute to the inadequacy of current therapeutic options for these variants. MicroRNAs (miRNA) are a class of small highly conserved regulatory endogenous non-coding RNA, which can alter the expression of genes encoding proteins and may play a role in the dysregulation of cellular pathways. Our goal was to improve the knowledge of the molecular pathogenesis of TNBC subgroups analyzing the miRNA expression profile, and to identify new prognostic and predictive biomarkers. We conducted a human miRNome analysis by TaqMan Low Density Array comparing different TNBC subtypes, defined by immunohistochemical basal markers EGFR and CK5/6. RT-qPCR confirmed differential expression of microRNAs. To inspect the function of the selected targets we perform Gene Ontology and KEGG enrichment analysis. We identified a single miRNA signature given by miR-135b expression level, which was strictly related to TNBC with basal-like phenotype. miR-135b target analysis revealed a role in the TGF-beta, WNT and ERBB pathways. A significant positive correlation was identified between neoplastic proliferative index and miR-135b expression. These findings confirm the oncogenic roles of miR-135b in the pathogenesis of TNBC expressing basal markers. A potential negative prognostic role of miR-135b overexpression might be related to the positive correlation with high proliferative index. Our study implies potential clinical applications: miR-135b could be a potential therapeutic target in basal-like TNBCs., Competing Interests: Competing Interests: The authors have declared that no competing interest exists.

Water monitoring requires expensive instrumentations and skilled technicians. In developing Countries as Africa, the severe economic restrictions and lack of technology make water safety monitoring approaches applied in developed Countries, still not sustainable. The need to develop new methods that are suitable, affordable, and sustainable in the African context is urgent. The simple, economic and rapid Micro Biological Survey (MBS) method does not require an equipped laboratory nor special instruments and skilled technicians, but it can be very useful for routine water analysis. The aim of this work was the application of the MBS method to evaluate the microbiological safety of different water sources and the effectiveness of different drinking water treatments in the Horn of Africa. The obtained results have proved that this method could be very helpful to monitor water safety before and after various purification treatments, with the aim to control waterborne diseases especially in developing Countries, whose population is the most exposed to these diseases. In addition, it has been proved that Moringa oleifera water treatment is ineffective in decreasing bacterial load of Eritrea water samples.

Work in the forestry industry is recognised internationally as high risk for occupational injuries and deaths. Forestry logging operations generally involve felling trees, skidding (moving trees and logs from the stump to the point of delivery), and transporting trees to mills. Loggers need to recognize the hazards associated with identifying dangerous (dead) trees, felling, delimbing, bucking, debarking, chipping, skidding, yarding, loading, unloading, and transporting logs. The purpose of this study was to determine the specific work conditions and job factors that workers identified as greatest risk for injury or death in logging operations in the intermountain region of Montana and Idaho. The study investigators recruited professional loggers through logging companies and professional logging associations in the states of Idaho and Montana. The professional loggers were asked to participate in focus group meetings regarding their perception of the association between logging tasks and logging injuries / deaths. One focus group session of 16 participants for 90 minutes was conducted and moderated by the investigators. The focus group moderator posed a series of questions designed to promote group discussion regarding logging practices and injuries. Injury data from workers' compensation reports was also shared with the participants and served as a basis for the focus group questions. Felling trees was often associated with injuries sustained from falling branches. Conventional chainsaw logging and skyline skidding were perceived as high-risk processes. Loggers indicated that being struck by the carriage attached to the skyline was a significant risk. Other risks identified included being hit by the butt of swinging trees when that became snagged or caught on tree stumps as they were being skidded up the slope. Workers identified working on steep slopes as well as working in below freezing temperatures as a major risk for injury. Physical fatigue while performing strenuous work for 8-10 hours was also identified as a risk for injury. The results of this study identified work conditions (extreme cold temperatures, steep incline of mountain slope), as well as job factors (felling trees, skyline skidding, driving logging trucks) that professional loggers perceived as contributing to increased risk for occupational related injury and death. The conditions and factors identified will be used to target interventions that reduce the risk of logging injuries in the future. Due to the remote nature of the logging work in this region, emergency first-aid training among the crewmembers was thought to be a critical part of logging safety. A culture promoting safety was also stressed for professional loggers.

The purpose of this study was to conduct and compare two ergonomic risk assessment methods often used in occupational health research and practice: the Strain Index (SI) and Occupational Repetitive Actions (OCRA) Checklist. Seven raters used the SI and OCRA Checklist to assess task-level physical exposures to the upper extremity of workers performing 21 cheese-manufacturing tasks. Of the total task exposures assessed with both methods, nearly half (49.1%) were classified as hazardous using the OCRA Checklist while 60.2% were classified as hazardous using the SI. Although the underlying injury risk characterization constructs of the SI and OCRA Checklist differ, the results indicated that the SI and OCRA Checklist often classified job tasks into similar risk categories. The differences in risk classifications determined by the SI and OCRA Checklist for job tasks were likely related to the definition of variables measured by these assessment methods as well as the complexity of tasks evaluated. By design, the SI is specific to the distal upper extremity while the OCRA Checklist accounts for the entire upper extremity including the shoulder. When conducting risk assessments of industrial work tasks, the choice of analysis tools should be based on the purpose of the assessment and the complexity of task functions. Both the SI and OCRA Checklist yield risk assessment ratings that are similar for cheese processing tasks.

Objectives: Commercial cow milking tasks, regardless of dairy size, have been documented in many regions of the world as strenuous work requiring high muscular effort, awkward positions, and task repetition. Large-herd dairies are common in the U.S., while Europe historically has mostly small-herd dairies. The objective of this study was to compare the upper limb muscle activity during milking tasks between workers at large-herd U.S. dairies and small-herd Italian dairies. This is the first international study directly comparing upper limb muscle activity among dairy workers from different countries using identical methods., Methods: Data were collected at 6 large-herd dairies in the U.S. region of Colorado and at 21 small-herd Italian dairies in the Lombardy region. Surface electromyography (sEMG) from the trapezius, anterior deltoid, biceps brachii, wrist flexors, and wrist extensors muscles was recorded from all participating workers (N = 65). Electromyography data were normalized to functional maximum voluntary contractions. Anthropometric measurements were also recorded., Results: Upper limb muscle activity was generally greater among workers in the large-herd U.S. dairies compared with small-herd Italian dairies. The amount of muscular rest as a percent of the work time was significantly greater among large-herd U.S. dairy workers., Conclusion: The differences revealed in sEMG and percent muscular rest among workers from the U.S. and Italy are likely due in part to differences in work processes adopted by fast-paced industrialized large-herd dairies compared with the slower, but sustained work processes performed at small-herd dairies.

Background: Triple Negative Breast Cancer (TNBC) accounts for 12-24% of all breast carcinomas, and shows worse prognosis compared to other breast cancer subtypes. Molecular studies demonstrated that TNBCs are a heterogeneous group of tumors with different clinical and pathologic features, prognosis, genetic-molecular alterations and treatment responsivity. The PI3K/AKT is a major pathway involved in the regulation of cell survival and proliferation, and is the most frequently altered pathway in breast cancer, apparently with different biologic impact on specific cancer subtypes. The most common genetic abnormality is represented by PIK3CA gene activating mutations, with an overall frequency of 20-40%. The aims of our study were to investigate PIK3CA gene mutations on a large series of TNBC, to perform a wider analysis on genetic alterations involving PI3K/AKT and BRAF/RAS/MAPK pathways and to correlate the results with clinical-pathologic data., Materials and Methods: PIK3CA mutation analysis was performed by using cobas® PIK3CA Mutation Test. EGFR, AKT1, BRAF, and KRAS genes were analyzed by sequencing. Immunohistochemistry was carried out to identify PTEN loss and to investigate for PI3K/AKT pathways components., Results: PIK3CA mutations were detected in 23.7% of TNBC, whereas no mutations were identified in EGFR, AKT1, BRAF, and KRAS genes. Moreover, we observed PTEN loss in 11.3% of tumors. Deregulation of PI3K/AKT pathways was revealed by consistent activation of pAKT and p-p44/42 MAPK in all PIK3CA mutated TNBC., Conclusions: Our data shows that PIK3CA mutations and PI3K/AKT pathway activation are common events in TNBC. A deeper investigation on specific TNBC genomic abnormalities might be helpful in order to select patients who would benefit from current targeted therapy strategies.

The purpose of this study was to characterize the inter-rater reliability of two physical exposure assessment methods of the upper extremity, the Strain Index (SI) and Occupational Repetitive Actions (OCRA) Checklist. These methods are commonly used in occupational health studies and by occupational health practitioners. Seven raters used the SI and OCRA Checklist to assess task-level physical exposures to the upper extremity of workers performing 21 cheese manufacturing tasks. Inter-rater reliability was characterized using a single-measure, agreement-based intraclass correlation coefficient (ICC). Inter-rater reliability of SI assessments was moderate to good (ICC = 0.59, 95% CI: 0.45-0.73), a similar finding to prior studies. Inter-rater reliability of OCRA Checklist assessments was excellent (ICC = 0.80, 95% CI: 0.70-0.89). Task complexity had a small, but non-significant, effect on inter-rater reliability SI and OCRA Checklist scores. Both the SI and OCRA Checklist assessments possess adequate inter-rater reliability for the purposes of occupational health research and practice. The OCRA Checklist inter-rater reliability scores were among the highest reported in the literature for semi-quantitative physical exposure assessment tools of the upper extremity. The OCRA Checklist however, required more training time and time to conduct the risk assessments compared to the SI., (Copyright © 2015 Elsevier Ltd and The Ergonomics Society. All rights reserved.)

Background: The region of Sardinia, Italy is known internationally for the production of cheeses made from ewe's milk. Although the use of automated milking equipment is available in ewe dairy operations, traditional hand milking is still performed on many ewe farms. The purpose of this study was to evaluate the prevalence of carpal tunnel syndrome (CTS) among farmers that manually milk ewes., Methods: Worker demographics, upper limb symptoms, and electrophysiologic studies were obtained on 76 ewe farmers recruited from a random sample of 109 in northern Sardinia. Characteristic hand symptoms and electrophysiologic studies were used in the case definition of CTS., Results: Of the 76 farmers evaluated, 42 (55.3%) fit the case definition of CTS in at least one hand. Assuming that all non-respondents did not have CTS, the CTS prevalence ratio would have been 38.5% (42/109) among the randomized sample of ewe farmers., Conclusions: These findings suggest that CTS is a significant occupational health issue for ewe farmers that continue the traditional methods of manual milking. The recent trend in automated ewe milking machines may help reduce the prevalence of CTS among the next generation of Italian ewe farmers., (Copyright © 2012 Wiley Periodicals, Inc.)

Aim: Different studies, carried out by us and others, have investigated the impact of carbon nanotubes (CNTs) in vitro and in animal models. To date, only a few studies have been performed on human cells ex vivo. There is also a lack of comparison between CNTs with varied functionalization and structural properties and their impact on different cell types., Materials & Methods: The present ex vivo human study focuses on the impact of a series of functionalized multiwalled CNTs on human T and B lymphocytes, natural killer (NK) cells and monocytes., Results: Smaller diameter nanotubes are internalized more efficiently. Viability assays displayed the absence of cytotoxicity of all multiwalled CNTs used. Activation assay demonstrated a strong effect on monocytes and NK cells., Conclusion: Our results, on human cells ex vivo, confirmed previous studies demonstrating appropriately functionalized CNTs are nontoxic. The effects on cell functionality were significant for the monocytes and NK cells. These findings encourage the possible use of CNTs for biomedical applications either as carriers of therapeutic molecules or as immune modulator systems.

Background: Patients on warfarin therapy undergo invasive and expensive checks for the coagulability of their blood. No information on coagulation levels is currently available between two controls., Methodology: A method was developed to determine warfarin in oral fluid by HPLC and fluorimetric detection. The chromatographic separation was performed at room temperature on a C-18 reversed-phase column, 65% PBS and 35% methanol mobile phase, flow rate 0.7 mL/min, injection volume 25 µL, excitation wavelength 310 nm, emission wavelength 400 nm., Findings: The method was free from interference and matrix effect, linear in the range 0.2-100 ng/mL, with a detection limit of 0.2 ng/mL. Its coefficient of variation was <3% for intra-day measurements and <5% for inter-day measurements. The average concentration of warfarin in the oral fluid of 50 patients was 2.5±1.6 ng/mL (range 0.8-7.6 ng/mL). Dosage was not correlated to INR (r = -0.03, p = 0.85) but positively correlated to warfarin concentration in the oral fluid (r = 0.39, p = 0.006). The correlation between warfarin concentration and pH in the oral fluid (r = 0.37, p = 0.009) confirmed the importance of pH in regulating the drug transfer from blood. A correlation between warfarin concentration in the oral fluid and INR was only found in samples with pH values ≥7.2 (r = 0.84, p = 0.004)., Conclusions: Warfarin diffuses from blood to oral fluid. The method allows to measure its concentration in this matrix and to analyze correlations with INR and other parameters.