Serum profiling by MALDI-TOF mass spectrometry as a diagnostic tool for domoic acid toxicosis in California sea lions

Background

There are currently no reliable markers of acute domoic acid toxicosis (DAT) for California sea lions. We investigated whether patterns of serum peptides could diagnose acute DAT. Serum peptides were analyzed by MALDI-TOF mass spectrometry from 107 sea lions (acute DAT n = 34; non-DAT n = 73). Artificial neural networks (ANN) were trained using MALDI-TOF data. Individual peaks and neural networks were qualified using an independent test set (n = 20).

Results

No single peak was a good classifier of acute DAT, and ANN models were the best predictors of acute DAT. Performance measures for a single median ANN were: sensitivity, 100%; specificity, 60%; positive predictive value, 71%; negative predictive value, 100%. When 101 ANNs were combined and allowed to vote for the outcome, the performance measures were: sensitivity, 30%; specificity, 100%; positive predictive value, 100%; negative predictive value, 59%.

Conclusions

These results suggest that MALDI-TOF peptide profiling and neural networks can perform either as a highly sensitive (100% negative predictive value) or a highly specific (100% positive predictive value) diagnostic tool for acute DAT. This also suggests that machine learning directed by populations of predictive models offer the ability to modulate the predictive effort into a specific type of error.     

Cell-free derivatives from mesenchymal stem cells are effective in wound therapy

AIM:To compare the efficacy of cell-free derivatives from Bone marrow derived human mesenchymal stem cells(hMSCs) in wound therapy.METHODS:hMSCs have been shown to play an important role in wound therapy.The present study sought to compare efficacy of hMSCs and cell-free derivatives of hMSCs,which may be clinically more relevant as they are easier to prepare,formulate and transport.hMSCs were isolated from human bone marrow and cultured.Multi lineage differentiation of hMSCs was performed to confirm their identity.The ability of hMSCs to migrate was evaluated using in vitro and in vivo migration assays.Cell lysates and conditioned medium concentrate was prepared from hMSCs(see Methods for details).Wounds were induced in mice and wound areas were measure before and after cell and cell-free derivative treatment.RNA and proteins were extracted from the skin and cytokine levels were measured.RESULTS:Co-culture of hMSCs with keratinocytes resulted in increased expression of CXCL-12(SDF1) and ENA78(CXCL-5) in the conditioned media indicating that the hMSCs can respond to signals from keratinocytes.Accelerated wound closure was observed when hMSCs were injected near the site of excisional wounds in athymic as well as NOD/SCID mice.Interestingly,cell-free lysates prepared from hMSCs were also effective in inducing accelerated wound closure and increased expression of SDF1 and CXCL-5 at the wound bed.Additionally,concentrated media from hMSCs as well as an emulsion containing lysates prepared from hMSCs was also found to be more effective in rapid re-epithelialization than fibroblasts or vehicle-alone control.Use of cell-free derivatives may help replace expensive wound care approaches including use of growth factors,epidermal/dermal substitutes,synthetic membranes,cytokines,and matrix components,and most importantly avoid transmission of pathogens from human and animal products.CONCLUSION:These results encourage development of derivatives of hMSCs for wound care and re-epithelialization applications.     

ACUTE EFFECTS OF MOVEMENT VELOCITY ON BLOOD LACTATE AND GROWTH HORMONE RESPONSES AFTER ECCENTRIC BENCH PRESS EXERCISE IN RESISTANCE-TRAINED MEN

This study aimed to compare the effects of different velocities of eccentric muscle actions on acute blood lactate and serum growth hormone (GH) concentrations following free weight bench press exercises performed by resistance-trained men. Sixteen healthy men were divided into two groups: slow eccentric velocity (SEV; n = 8) and fast eccentric velocity (FEV; n = 8). Both groups performed four sets of eight eccentric repetitions at an intensity of 70% of their one repetition maximum eccentric (1RMecc) test, with 2-minute rest intervals between sets. The eccentric velocity was controlled to 3 seconds per range of motion for SEV and 0.5 seconds for the FEV group. There was a significant difference (P < 0.001) in the kinetics of blood lactate removal (at 3, 6, 9, 15, and 20 min) and higher mean values for peak blood lactate (P = 0.001) for the SEV group (9.1 ± 0.5 mM) compared to the FEV group (6.1 ± 0.4 mM). Additionally, serum GH concentrations were significantly higher (P < 0.001) at 15 minutes after bench press exercise in the SEV group (1.7 ± 0.6 ng · mL⁻¹) relative to the FEV group (0.1 ± 0.0 ng · mL⁻¹). In conclusion, the velocity of eccentric muscle action influences acute responses following bench press exercises performed by resistance-trained men using a slow velocity resulting in a greater metabolic stress and hormone response.     

Cytotoxicity induced by grape seed proanthocyanidins: Role of nitric oxide

Grape seed proanthocyanidin extract (GPSE) at high doses has been shown to exhibit cytotoxicity that is associated with increased apoptotic cell death. Nitric oxide (NO), being a regulator of apoptosis, can be increased in production by the administration of GSPE. In a chick cardiomyocyte study, we demonstrated that high-dose (500 μg/ml) GSPE produces a significantly high level of NO that contributes to increased apoptotic cell death detected by propidium iodide and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. It is also associated with the depletion of intracellular glutathione (GSH), probably due to increased consumption by NO with the formation of S-nitrosoglutathione. Co-treatment with L-NAME, a NO synthase inhibitor, results in reduction of NO and apoptotic cell death. The decline in reduced GSH/oxidized GSH (GSSG) ratio is also reversed. N-Acetylcysteine, a thiol compound that reacts directly with NO, can reduce the increased NO generation and reverse the decreased GSH/GSSG ratio, thereby attenuating the cytotoxicity induced by high-dose GSPE. Taken together, these results suggest that endogenous NO synthase (NOS) activation and excessive NO production play a key role in the pathogenesis of high-dose GSPE-induced cytotoxicity.     

The mutation rates of di-, tri- and tetranucleotide repeats in Drosophila melanogaster

In a recent study, we reported that the combined average mutation rate of 10 di-, 6 tri-, and 8 tetranucleotide repeats in Drosophila melanogaster was 6.3 x 10(-6) mutations per locus per generation, a rate substantially below that of microsatellite repeat units in mammals studied to date (range = 10(-2)-10(-5) per locus per generation). To obtain a more precise estimate of mutation rate for dinucleotide repeat motifs alone, we assayed 39 new dinucleotide repeat microsatellite loci in the mutation accumulation lines from our earlier study. Our estimate of mutation rate for a total of 49 dinucleotide repeats is 9.3 x 10(-6) per locus per generation, only slightly higher than the estimate from our earlier study. We also estimated the relative difference in microsatellite mutation rate among di-, tri-, and tetranucleotide repeats in the genome of D. melanogaster using a method based on population variation, and we found that tri- and tetranucleotide repeats mutate at rates 6.4 and 8.4 times slower than that of dinucleotide repeats, respectively. The slower mutation rates of tri- and tetranucleotide repeats appear to be associated with a relatively short repeat unit length of these repeat motifs in the genome of D. melanogaster. A positive correlation between repeat unit length and allelic variation suggests that mutation rate increases as the repeat unit lengths of microsatellites increase.      

Morphology of the endometrial microvasculature during early placentation in the rat

Summary The morphology of the uterine microvasculature during early placentation was investigated by light microscopy, scanning electron microscopy of microvascular corrosion casts and transmission electron microscopy in rats 26 and 50 h after initiation of implantation. Increased vascular permeability at implantation sites was observed as a positive blue-dye test, spacing of vessels, and as localized extravasations of resin from postcapillary venules in the center of the endometrium. The subepithelial capillary plexus in the primary decidual zone adjacent to the blastocyst was shut down 50 h after initiation of implantation, most probably due to swelling of the metabolically activated endothelium and volume expansion of the decidual cells. This phenomenon coincided with the mesometrial orientation of the inner cell mass of the blastocyst; it may be a uterine mechanism to direct the ectoplacental cone toward the patent vessels in the mesometrial portion of the uterus. The remaining vessels at implantation sites were generally fewer, larger in diameter, more irregular in caliber, and more uniformly oriented along the implantation axis than their counterparts at inter-implantation sites. No vascular sprouts were observed during the interval studied.     

Proffered papers 14.00–15.00 Tuesday 16 September 2003

Background  ERCP-directed brush cytology is used to sample lesions of the pancreatic and biliary ducts and the ampulla of Vater. With conventional preparations, the sensitivity and specificity range from 44% to 63% and 80% to 98%, respectively, and increased N : C ratio, nuclear molding and loss of honeycombing are reliable features of malignancy. The performance and morphology of specimens prepared by ThinPrep, a liquid-based cytology technique is mostly unknown.
Methods  The laboratory information system was searched for all cases prepared by ThinPrep. Patient disease classification of benign or malignant was determined by linkage with the provincial cancer registry and was the gold standard against which sensitivity, specificity, positive (PPV) and negative predictive values (NPV) were calculated. True positives and negatives were reviewed to identify predictive cytomorphologic features.
Results  Between 1996 and 2001, there were 149 ThinPrep specimens; 55 (37%) were reported as positive for malignancy and 94 (63%) as negative. Disease was classified as malignant in 86 (58%) patients and benign in 63 (42%). There were 42 false negative, 11 false positive, 52 true negative, and 44 true positive cytology results. Sensitivity was 51.2% (CI; 40.2 : 62.0), specificity 82.5% (CI; 70.5 : 90.6), and PPV and NPV 80.0% (CI; 66.6 : 89) and 55.3% (CI; 44.7 : 65.5), respectively. Cell groups with crowded, enlarged, irregular nuclei and nuclear features of vesicular chromatin and large, multiple irregular nucleoli correlated with malignant disease, while monolayered sheets of uniform columnar cells, regular nuclei and a finely granular chromatin correlated with benign disease.
Conclusions  The performance of ThinPrep brushings from this anatomic site equals conventional preparations. Cytomorphologic features of malignancy are more frequent and pronounced with ThinPrep.     

Biofloc Technology: Emerging Microbial Biotechnology for the Improvement of Aquaculture Productivity

With the significant increases in the human population, global aquaculture has undergone a great increase during the last decade. The management of optimum conditions for fish production, which are entirely based on the physicochemical and biological qualities of water, plays a vital role in the prompt aquaculture growth. Therefore, focusing on research that highlights the understanding of water quality and breeding systems’ stability is very important. The biofloc technology (BFT) is a system that maximizes aquaculture productivity by using microbial biotechnology to increase the efficacy and utilization of fish feeds, where toxic materials such as nitrogen components are treated and converted to a useful product, like a protein for using as supplementary feeds to the fish and crustaceans. Thus, biofloc is an excellent technology used to develop the aquaculture system under limited or zero water exchange with high fish stocking density, strong aeration, and biota. This review is highlighted on biofloc composition and mechanism of system work, especially the optimization of water quality and treatment of ammonium wastes. In addition, the advantages and disadvantages of the BFT system have been explained. Finally, the importance of contemporary research on biofloc systems as a figure of microbial biotechnology has been emphasized with arguments for developing this system for better production of aquaculture with limited natural resources of water.Key words: biofloc, BFT, aquaculture, microbes, water quality, wastes