In vitro assessment of the efficacy of protein exudates from seeds against Haemonchus contortus.

Nematodes develop resistance to the most common commercially available drugs. The aim of this study was to identify and evaluate the action of protein exudates from Mimosa caesalpiniifolia, Leucaena leucocephala, Acacia mangium, and Stylosanthes capitata seeds on the gastrointestinal nematode Haemonchus contortus. The exuded proteins were precipitated, dialyzed, lyophilized, and assessed for their effect on egg hatching and artificial larval exsheathment inhibition. Proteome analysis of the protein extracts was also performed. Although no egg-hatching inhibition was observed, all exudates showed efficacy in inhibiting the larval exsheathment of H. contortus larvae with an EC50 varying from 0.61 to 0.26 mg P mL-1. Proteomic analysis revealed the presence of proteases, protease inhibitors, chitinases, and lectins among other proteins in the exudates. Most of the exuded proteins belong to the oxidative stress/plant defense and energy/carbohydrate metabolism functional clusters. This study concluded that the bioactive proteins from different classes exuded by seeds of M. caesalpiniifolia, L. leucocephala, A. mangium, and S. capitata show stage-specific inhibition against H. contortus.

A novel method for determination and quantification of 4-methyloctanoic and 4-methylnonanoic acids in mutton by hollow fiber supported liquid membrane extraction coupled with gas chromatography.

4-Methyloctanoic acid (MOA) and 4-methylnonanoic acid (MNA) are the main compounds responsible for "sweaty" odor of mutton. A novel method for their determination has been developed and validated. Hollow fiber supported liquid membrane (HF-SLM) was applied to selectively extract MOA and MNA prior to gas chromatography (GC) analysis. For HF-SLM, the donor outside the fiber was the acidified supernatant (pH 4) from aqueous mutton slurry. Liquid membrane was 5% tri-n-octylphoshphine oxide in di-n-hexyl ether and 0.3M NaOH aqueous solution filled in the lumen of the fiber was used as the acceptor. The extraction last for 4h. After acidification with HCl, the acceptor was directly analyzed by GC. Importantly, HF-SLM provided high enrichment factors for MOA (133) and MNA (116). The method developed had low detection limits of 0.0007-0.0015 mg/kg, good linearity (R²>0.9956), reasonable recovery (88.54-122.13%), satisfactory intra-assay (7.83-9.73%) and inter-assay (15.68-16.14%) precision.

Pleural fluid C-reactive protein contributes to the diagnosis and assessment of severity of parapneumonic effusions.

BACKGROUND AND AIMS: Prompt identification of parapneumonic effusions has immediate therapeutic benefits. We aimed to assess whether C-reactive protein (CRP) and routine biochemistries in pleural fluid are accurate markers of parapneumonic effusions, and to evaluate their properties as indicators for drainage (complicated parapneumonic effusion). METHODS: A retrospective review of 340 non-purulent parapneumonic effusions and 1,659 non-parapneumonic exudates from a single center was performed and the discriminative properties of pleural fluid routine biochemistries and, when available, CRP were evaluated. CRP, along with classical fluid parameters, was also applied to classify patients as having complicated or uncomplicated parapneumonic effusions. ROC analysis established the threshold of CRP for discriminating between groups. RESULTS: Pleural fluids with neutrophilic predominance and CRP levels >45 mg/dL were most likely to be parapneumonic in origin (likelihood ratio=7.7). When attempting to differentiate non-purulent complicated from uncomplicated effusions, a CRP >100mg/L had the same performance characteristics (area under the curve=0.81) as the widely accepted biochemistries pH and glucose. Combinations of CRP with pH or glucose resulted in incrementally discriminating values, pertaining to either sensitivity (75-80%) or specificity (97%), for complicated effusions. CONCLUSION: Pleural fluid CRP may be a useful adjunctive test in pleural effusions, both as a marker of parapneumonics and, particularly, as a differentiator between complicated and uncomplicated effusions.

Exudate assessment and management.

Good wound management must involve an holistic approach to care; without considering the whole person, appropriate management might not be as good as it could be. In a time of austerity, it is important that money is spent appropriately on the correct wound-management technologies. Exudate assessment and management are a vital part of wound management. In this article, the focus will be on exploring the nature of exudate and tools available to evaluate exudate. Finally, suggestions will be made on the management of exudate.

A comparative analysis of the biochemical parameters used to distinguish between pleural exudates and transudates.

OBJECTIVE: The aim of the present study was to compare the various parameters used to identify exudates. METHODOLOGY: The study included 255 patients with pleural effusions. According to aetiological diagnosis, 105 pleural effusions were labelled as transudates and 150 were labelled as exudates. RESULTS: Using the criteria of Light et al., 94.5% of the effusions were correctly classified, yielding a sensitivity and specificity of 99.3% and 87.6%, respectively. Use of the pleural fluid/serum bilirubin ratio produced results of 92.9%, 90.7%, and 96.2%, respectively. Using pleural fluid cholesterol level yielded results of 95.7%, 95.3%, and 96.2%, respectively. When the combination of pleural fluid cholesterol level and lactate dehydrogenase (LDH) level was used, the specificity and accuracy were found to be higher than that using the criteria of Light et al. We found that there was no significant difference among the parameters with respect to accuracy. CONCLUSION: When the accuracy and cost are considered, differentiation of pleural exudates and transudates can be achieved only by pleural fluid cholesterol level or LDH level; and when two parameters were used together, the accuracy and specificity were higher than that using the criteria of Light et al.

Assessment of postmortem blood alcohol concentrations by ethanol levels measured in fluids from putrefactive blisters.

It was the aim of the present study to investigate whether the postmortem blood alcohol concentration (BAC) can be assessed by means of ethanol values measured in fluids from putrefactive blisters. Fluids from one or two putrefactive blisters were collected in 45 putrefied corpses (postmortem interval: 3-23 days, graduation of the degree of decomposition into four stages) and compared with femoral vein blood or femoral muscle. Ethanol and so-called putrefactive alcohols were determined in all samples by double GC-measurements. The results were converted to the medium water content of blood of 80%. The investigations revealed a significant correlation between the ethanol values of femoral blood/muscle and putrefactive blister fluid (PBF) (r = 0.725, P < 0.001). With the exception of one case, PBF values of 0.11% or more (n = 11) corresponded to minimum blood/muscle concentrations of 0.085%; PBF values of more than 0.15% (n = 8) corresponded to blood/muscle levels of at least 0.159%. Regression analysis revealed the formula BAC (in %) = 0.785 X ethanol (PBF) +0.011. Relevant concentrations of putrefactive alcohols such as 1- and 2-propanol and l-butanol only occurred in cases with far advanced decomposition. Ethanol levels determined in fluids from different putrefactive blisters of the same body also demonstrated a significant correlation (r = 0.791, P < 0.001). Ethanol concentrations determined in PBF must be judged with caution, but PBF values > 0.15% point to corresponding blood levels with a high degree of probability, at least where advanced putrefaction is excluded.

Sensitivity of lungs of aging Fischer 344 rats to ozone: assessment by bronchoalveolar lavage.

Biological effects indicators in bronchoalveolar lavage fluid were studied in Fischer 344 rats of different ages after exposure to 0.4-0.8 ppm ozone for periods of 2-6 h on a single day or on 4 consecutive days. The magnitude of alveolar protein transudation induced by ozone was not different between age groups, but the interindividual variability of protein changes was higher in senescent (24-mo-old) rats. By comparison to juvenile (2-mo-old) and adult (9-mo-old) rats, senescent animals had higher increases of interleukin-6 (up to 10-fold higher) and N-acetyl-beta-D-glucosaminidase (NAGA; 2-fold higher) in lung lavage after ozone. Ascorbic acid was lower in lungs of senescent rats (one-half of juvenile values), and acute ozone exposure brought a further decrease in lung ascorbate. Whereas alveolar protein transudation was attenuated after ozone exposure on 4 days, persistent elevation of NAGA in senescent rats suggested only partial adaptation. Injection of endotoxin did not modify the patterns of effects. Incorporation of 18O-ozone into macrophages and surfactant was not different between age groups, indicating that the magnified biological responses in senescent rats were not dominated by differences in internal dose of ozone. The results indicate that senescent rats respond differently than juvenile and adult rats to lung injury.

Cyst fluid analysis in the differential diagnosis of pancreatic cysts: a new approach to the preoperative assessment of pancreatic cystic lesions.

Pancreatic cysts include pseudocysts, serous tumors, and mucinous tumors, some of which are malignant. The clinical and radiologic features of these lesions may not permit a preoperative diagnosis. Analysis of percutaneous cyst fluid aspirates for cytologic findings, tumor markers, enzymes, and viscosity has been proposed as an aid to differential diagnosis. The combination of cytologic findings, carcinoembryonic antigen level, and viscosity will usually differentiate mucinous from nonmucinous cysts, whereas cytologic findings and levels of tumor markers CA 15-3 and CA 72-4 are useful to predict malignancy. Pseudocysts can be identified by a combination of cytologic features; levels of tumor markers, including NB/70K; and levels of enzymes, including amylase isoenzymes and leukocyte esterase. Serous tumors are a diagnosis of exclusion, although in 50% of cases the cytologic findings will indicate a serous tumor. When compared with conventional methods of differentiating pancreatic cysts, cyst fluid analysis is a promising new approach to the evaluation of these diagnostically difficult lesions.

Comparative assessment of DNA analysis in effusions by image analysis and flow cytometry.

Cytologic evaluation of body cavity fluids is useful to detect malignancy within the pleural and peritoneal spaces. A definitive diagnosis cannot always be made on cytologic evaluation alone. As malignant processes may show abnormal DNA content, DNA analysis of effusions may be useful. Therefore, we determined the DNA content of 37 effusions by flow cytometry (FC) and image analysis (IA) using the CAS 200. Of the 37 fluids evaluated, 18 were cytologically malignant, 15 benign, and four atypical. Overall, 22 fluids (60%) showed concordance between FC and IA. None of the benign fluids were aneuploid. All showed diploid histograms or diploidy with increased proliferating cells. Three of four atypical fluids had increased proliferating cells by either FC or IA, whereas one was diploid by both methods. Aneuploidy was detected in 13 malignant fluids: five were aneuploid by both methods and eight by only one method. IA identified aneuploidy in five of those eight cases, while three were identified by FC. Three of the cytologically malignant fluids were diploid by both methods, and two showed increased proliferating cells by IA and diploidy by FC. The specificity of both methods was 100%. However, the sensitivity of identifying a malignant fluid by aneuploidy is low, 44% for FC and 55% for IA. IA appears to identify small aneuploid populations more frequently than FC. The detection of aneuploidy in effusions is highly suggestive of malignancy, and the combination of both techniques gives the highest detection rate (72%). However, neither are as sensitive as traditional cytologic evaluation with the occasional use of additional histochemical stains.