Comprehensive proteomic analysis of Penicillium verrucosum.

Mass spectrometric identification of proteins in species lacking validated sequence information is a major problem in veterinary science. In the present study, we used ochratoxin A producing Penicillium verrucosum to identify and quantitatively analyze proteins of an organism with yet no protein information available. The work presented here aimed to provide a comprehensive protein identification of P. verrucosum using shotgun proteomics. We were able to identify 3631 proteins in an "ab initio" translated database from DNA sequences of P. verrucosum. Additionally, a sequential window acquisition of all theoretical fragment-ion spectra analysis was done to find differentially regulated proteins at two different time points of the growth curve. We compared the proteins at the beginning (day 3) and at the end of the log phase (day 12).

Indonesian wild honey authenticity analysis using attenuated total reflectance-fourier transform infrared (ATR-FTIR) spectroscopy combined with multivariate statistical techniques.

Wild honeys in Indonesia are still widely believed to be good for health with high economic value. This honey is naturally produced by Apisdorsata bee. In this study, authentication analysis by classification and discrimination of attenuated total reflectance-fourier infrared spectroscopy (ATR-FTIR) spectra was conducted on several wild honeys from various places in Indonesia (n = 186) which then compared to adulterated honey contained commercial sugars of aren (Arenga pinnata), coconut, and cane sugar at 10-50% concentration (n = 57). Combination of spectra measurement at 4,000-650 cm-1 with Chemometric technique by several multivariate analyses resulted in visualization of honey grouping, classification, and regression model that differentiate these honeys, both partial and overall. Principle component analysis multivariate analysis was able to visualize the differentiation of adulterated honey from the authentic ones. Discriminant analysis, a supervised classification technique, was used to differentiate the fake from the authentic honey among those from various origins at wave number range of 4000-800 cm-1 with performance index of 91,8, 90.32-100% sensitivity, and 95. 70-100% specificity. Partial least-squares analysis was used to build a model provided quantitative results of commercial sugars content in honey allegedly added during adulteration. Authentic honeys had commercial sugars content less than 10% with R2 of aren, coconut, and cane sugar of 0.9995, 0.9980 and 0.9998, respectively, with their predictive R2 values of 0.9977, 0.9983 and 0.9946, respectively.

Physicochemical Properties of Gelatin Extracted from Buffalo Hide Pretreated with Different Acids.

The acid pretreatment of collagen molecules disrupts their crosslinks and assists in the release of acid-soluble proteins, fats, and other components. Generally, to achieve optimum extraction efficiency, strong acids may be used at a lower acid concentration compared to weak acids. This study aimed to determine the yield and physicochemical properties of gelatins extracted from buffalo hides pretreated with different acids. Hides were extracted with hydrochloric, citric, and acetic acids at concentrations of 0.3, 0.6, 0.9, 1.2, and 1.5 M. A completely randomized design and the least significant difference test were used in the experimental design, and all measurements were performed in triplicate. The highest yield (29.17%) was obtained from pretreatment with 0.9 M HCl. The gel strength did not differ significantly (p>0.05) according to acid type (280.26-259.62 g Bloom), and the highest viscosity was obtained from the 0.6 M citric acid pretreatment. All the gelatins contained α- and ß-chain components and several degraded peptides (24-66 kDa). The color and Fourier-transform infrared spectrum of the gelatin extracted using 0.9 M HCl were similar to those of commercial bovine skin gelatin. In general, the physicochemical properties of the gelatin complied with the industry standard set by the Gelatin Manufacturers Institute of America, revealing that buffalo hide could serve as a potential alternative source of gelatin.

Coronary artery disease in patients at low risk--apolipoprotein AI as an independent risk factor.

Many risk factors for coronary artery disease (CAD) have been established by large epidemiological studies. However, some patients without the major risk factors still develop disease. Preliminary analysis of individuals referred for angiography, who had no major risk factors associated with CAD, indicated that apolipoprotein-AI (apoAI) was significantly lower in patients with positive angiograms. The hypothesis that apoAI was an independent risk factor for CAD in low risk populations was put forth. One thousand and seventy-five consecutive patients underwent angiography, lipid analysis, and completed a risk factor questionnaire. Individuals with total cholesterol<5.2 mmol/l, high density lipoprotein (HDL)-cholesterol>0.9 mmol/l, systolic blood pressure<140 mmHg and diastolic blood pressure<90 mmHg, no diabetes, and no family history of premature CAD in first degree relatives were selected. Fifty-four patients met these selection criteria, 29 having positive evidence of CAD and 25 with no evidence of disease. Multivariate analysis revealed that, after adjusting for age and gender, serum apoAI level was the only variable predictive of CAD. This effect was independent of HDL cholesterol level and fractional esterification rate of HDL (FER(HDL)). These results point to an important role for apoAI in the atherogenic process, particularly in populations with no major CAD risk factors. Decreased levels of apoAI or LpAI may initiate atherosclerosis in a highly selected group of low risk patients.

Exclusion of noninfectious medical waste from the contaminated waste stream.

The purpose of this study was to determine whether any waste was labeled incorrectly as infectious or contained material that could be recycled. Waste generated in preparation for surgery was separated and weighed. Of 530 lbs of operating room waste, 40 lbs were noninfectious, largely plastic and paper. The results of the present study suggest a segment of waste can be removed from the contaminated stream, potentially reducing hospital costs and improving our environment.

Mutagenesis by chemical agents in V79 chinese hamster cells: a review and analysis of the literature. A report of the Gene-Tox Program.

The report reviews and evaluates the current literature (about 125 primary publications) on chemically induced specific locus mutations in the V79 Chinese hamster lung cell line. The V79 cell is convenient to use for mutagenesis studies since it has a rapid growth rate, high plating efficiency, and a stable karyotype. Mutation can be easily measured at either the hypoxanthine-guanine phosphoribosyl transferase or the Na+/K+ ATPase locus, both of which have been well characterized. Other less-studied markers are also described. We discuss the protocols for quantitative mutation studies including measurements of cytotoxicity, mutant expression times, mutant selection agents, cell densities during selection, and the stability and verification of mutant phenotypes. Mutations in the V79 cells by chemicals that require activation can be tested after their metabolism by cell homogenates or by intact cells, and the results with each type of activation are compared. For purposes of analysis, we classified a compound as mutagenic if it induced a mutation frequency that is at least 3 times higher than the spontaneous mutant frequency reported for that specific experiment. By this criterion two-thirds of the chemicals analyzed were mutagenic--; 11% with and 55% without metabolic activation. Of the 191 chemicals examined; 119 were polycyclic aromatic hydrocarbons; 25 were nitro or nitroso compounds, 9 were alkyl halides; 7 were purine or pyrimidine derivatives and the remaining 31 were from other chemical classes. We also defined mutagenic potency as the concentration of a compound that increases the mutant frequency by 10 times the spontaneous frequency. Mutagenic potencies of the compounds examined varied over a range of 5 X 10(6). We have also found large interlaboratory variations in the mutagenic potencies. Such variation in potency could be reduced by normalizing the results to a standard mutagen such as N-methyl-N'-nitro-N-nitrosoguanidine. The role of the V79 assay in mutagenicity and carcinogenicity testing is discussed and recommendations are suggested for future investigation.

Species comparison of protein synthesis accuracy.

The average accuracy of protein synthesis in reticulocytes from several mammalian species does not correlate with longevity potential from 13 to 90 years. Isoleucine incorporation into highly purified hemoglobin chains which contain no genetically coded isoleucine was used as a direct test of protein synthesis accuracy. Since isoleucine can be incorporated into these molecules by mutations in a few cells as well as errors in most cells, the constant level of isoleucine substitution may also show that the mutation rates are not dramatically different among these species. Isoleucine substitutions in hemoglobin can be used to estimate mutations only above the level of errors, which may be as low as 1/1,000,000, but the probability of seeing a few mutant clones at any time is dependent on the number of stem cells producing reticulocytes. The number of stem cells being expressed is a reflection of the number of cell divisions per clone. If the number of cell divisions per clone is 30 or less, then isoleucine substitutions would increase when the mutation accumulation rose above 30 per million for the mutation to isoleucine at any position in the alpha or beta chain.