Development of technologies aiding large-tissue engineering

Apoptosis associated oligonucleosomal fragmentation of DNA can result from the activation of endonucleases that exhibit different pH optima and are either sensitive or insensitive to divalent cations. DNA fragmentation due to activation of cation sensitive endonucleases occurs in the absence of a change in intracellular pH whereas intracellular acidification is a feature of apoptosis characterized by activation of cation insensitive acidic endonuclease. We have reported earlier that somatostatin (SST) induced DNA fragmentation and apoptosis is signaled in a receptor subtype selective manner uniquely via human somatostatin receptor subtype 3 (hSSTR3). In the present study we investigated the pH dependence and cation sensitivity of endonuclease induced in hSSTR3 expressing CHO-K1 cells by the SST agonist octreotide (OCT) and its effect on intracellular pH. We show that OCT induced apoptosis is associated with selective stimulation of a divalent cation insensitive acidic endonuclease. The intracellular pH of of cells undergoing OCT induced apoptosis was 0.9 pH units lower than that of control cells. The effect of OCT on endonuclease and pH was inhibited by orthovanadate as well as by pretreatment with pertussis toxin, suggesting that hSSTR3 initiated cytotoxic signaling is protein tyrosine phosphatase mediated and is G protein dependent. These findings suggest that intracellular acidification and activation of acidic endonuclease mediate wild type p53 associated apoptosis signaled by hormones acting via G protein coupled receptors.     

Comparative evaluation of FUNGITEST and broth microdilution methods for antifungal drug susceptibility testing of Candida species and Cryptococcus neoformans

The FUNGITEST method (Sanofi Diagnostics Pasteur, Paris, France) is a microplate-based procedure for the breakpoint testing of six antifungal agents (amphotericin B, flucytosine, fluconazole, itraconazole, ketoconazole, and miconazole). We compared the FUNGITEST method with a broth microdilution test, performed according to National Committee for Clinical Laboratory Standards document M27-A guidelines, for determining the in vitro susceptibilities of 180 isolates of Candida spp. (50 C. albicans, 50 C. glabrata, 10 C. kefyr, 20 C. krusei, 10 C. lusitaniae, 20 C. parapsilosis, and 20 C. tropicalis isolates) and 20 isolates of Cryptococcus neoformans. Overall, there was 100% agreement between the methods for amphotericin B, 95% agreement for flucytosine, 84% agreement for miconazole, 83% agreement for itraconazole, 77% agreement for ketoconazole, and 76% agreement for fluconazole. The overall agreement between the methods exceeded 80% for all species tested with the exception of C. glabrata (71% agreement). The poorest agreement between the results for individual agents was seen with C. glabrata (38% for fluconazole, 44% for ketoconazole, and 56% for itraconazole) and C. tropicalis (50% for miconazole). The FUNGITEST method misclassified as susceptible 2 of 12 (16.6%) fluconazole-resistant isolates, 2 of 10 (20%) itraconazole-resistant isolates, and 4 of 8 (50%) ketoconazole-resistant isolates of several Candida spp. Further development of the FUNGITEST procedure will be required before it can be recommended as an alternative method for the susceptibility testing of Candida spp. or C. neoformans.     

Oxidation and quality of soybean oil: A preliminary study of the anisidine test

The anisidine test, a measure of secondary oxidation products in glyceride oils, was applied to a number of soybean salad oils processed from sound and damaged soybeans. A highly significant correlation (−0.68) was found between the anisidine values of salad oils from sound soybeans and their flavor scores. Multiple correlations between flavor scores, anisidine, and peroxide values yielded a correlation of 0.81 and provided a method for predicting the initial flavor scores of sound soybean salad oils. Similar data for oils from damaged beans gave a highly significant, but lower, correlation (−0.65). Comparative studies indicated that sound crude oils usually contain lower levels of oxidation products than damaged crude. Oxidation in both sound and damaged crudes increased roughly in proportion to iron content. Reproducibility of the test and the effects of hydrogenation, accelerated storage, and fluorescent light on anisidine values were studied. Analysis of damaged oils before and after deodorization showed that little, if any, reduction of anisidine value occurred. Deodorization of sound oils, however, lowered anisidine values. In comparison with damaged oils, the anisidine values of sound oils were lower at comparable stages of processing. The poor quality of damaged soybean oil was substantiated by organoleptic evaluations. Flavor scores of oils given special processing treatments increased as anisidine values decreased.     

Flavor and oxidative stability of hydrogenated and unhydrogenated soybean oils: Effects of antioxidants

Flavor and oxidative stabilities were studied by organoleptic evaluation and chemical analysis of three different samples of soybean oil: unhydrogented (I); hydrogenated with nickel catalyst (II); and hydrogenated with copper-chromium catalyst (III). Analyses for these oils were: I II III iodine Value 138 109 113 Linolenate, % 8.3 3.3 0.4 Each oil was deodorized with the addition of either citric acid alone or citric acid plus BHA and BHT antioxidants. Addition of antioxidants did not improve the flavor stabilities of the oils in accelerated storage tests but did improve the flavor stabilities of II and III in light exposure tests. All three oils that received the same additive treatment had equivalent flavor stability in both accelerated storage and light exposure tests. However, both hydrogenation and antioxidant treatment improved oxidative stability as measured by the Active Oxygen Method. There was good correlation between flavor score and the logarithm of the peroxide value determined at the time of tasting. Presented at the AOCS Meeting, New York, May 1977.     

Flavor stability of soybean oil based on induction periods for the formation of volatile compounds by gas chromatography

Although previous research showed that volatile compounds detected by gas chromatography (GC) correlated well with flavor scores, no instrumental or chemical method has been available to predict flavor stability of vegetable oils reliably. A direct GC method was tested to predict flavor stability of soybean oil by measuring induction periods based on the time required for rapid formation of volatile compounds. By this technique, induction periods of 9, 5 and 0 days were obtained with oils containing a combination of tertiary butylhydroquinone (TBHQ) and citric acid (CA), CA only and no additives, respectively. Addition of methyl silicone to the oils containing CA or CA+TBHQ did not increase their stability. Prominent peaks identified by gas chromatography-mass spectrometry included: pentane, hexanal, 2-heptanal, 2,4-heptadienal, 2-decenal and 2,4-decadienal. Measures of total volatile compounds, pentane and 2,4-decadienal were best related to deteriorative changes. High correlation coefficients were obtained between individual and total volatiles with flavor scores. This study showed that flavor stability of oils can be predicted by determining induction periods based on the formation of volatile compounds.     

Performance evaluation of hexane-extracted oils from genetically modified soybeans

Soybeans produced by induced mutation breeding and hybridization were cracked, flaked and hexane-extracted, and the recovered crude oils were processed to finished edible oils by laboratory simulations of commercial oil-processing procedures. Three lines yielded oils containing 1.7, 1.9 and 2.5% linolenic acid. These low-linolenic acid oils were evaluated along with oil extracted from the cultivar Hardin, grown at the same time and location, and they were processed at the same time. The oil from Hardin contained 6.5% linolenic acid. Low-linolenic acid oils showed improved flavor stability in accelerated storage tests after 8 d in the dark at 60°C and after 8h at 7500 lux at 30°C, conditions generally considered in stress testing. Room odor testing indicated that the low-linolenic oils showed significantly lower fishy odor after 1 h at 190°C and lower acrid/pungent odor after 5 h. Potatoes were fried in the oils at 190°C after 5, 10 and 15 h of use. Overall flavor quality of the potatoes fried in the low-linolenic oils was good and significantly better after all time periods than that of potatoes fried in the standard oil. No fishy flavors were perceived with potatoes fried in the low-linolenic oils. Total volatile and polar compound content of all heated oils increased with frying hours, with no significant differences observed. After 15 h of frying, the free fatty acid content in all oils remained below 0.3%. Lowering the linolenic acid content of soybean oil by breeding was particularly beneficial for improved oil quality during cooking and frying. Flavor quality of fried foods was enhanced with these low-linolenic acid oils.     

Flavor and oxidative stability of continuously hydrogenated soybean oils

Soybean oil was partially hydrogenated in a continuous system with copper and nickel catalysts. The hydrogenated products were evaluated for flavor and oxidative stability. Processing conditions were varied to produce oils of linolenate contents between 0.4 and 2.7%, as follows: oil flow, 0.6–2.2 liters/hr; reaction temperature, 180–220 C; hydrogen pressure, 100–525 psig, and catalyst concentration, 0.5–1% copper catalyst or 0.1% nickel catalyst.Trans unsaturation varied from 8 to 20% with copper catalyst and from 15.0 to 27% with nickel catalyst. Linolenate selectivity was 9 with copper catalyst and 2 with nickel catalyst. Flavor evaluation of finished oils containing 0.01% citric acid (CA), appraised initially and after accelerated storage at 60 C, showed no significant difference between hydrogenated oils and nonhydrogenated oil. However, peroxide values and oxidative stability showed that hydrogenated oils were more stable than the unhydrogenated oil. CA+TBHQ (tertiary butylhydroquinone) significantly improved the oxidative stability of test oils over oils with CA only, but flavor scores showed no improvement. Dimethylpolysiloxane (MS) had no effect on either flavor or oxidative stability of the oils.     

Arthroscopic Bankart repair with the Suretac device. Part I: Clinical observations

Although arthroscopic Bankart repair has become an accepted surgical stabilization technique for anterior shoulder instability, the failure rate remains unacceptably high. Little information is available concerning healing of the Bankart repair. The purpose of this article is to clarify this issue by analyzing a cohort of 15 patients who underwent a "second-look" arthroscopy to evaluate and treat pain or recurrent instability following arthroscopic Bankart repair with the Suretac device (Acufex Microsurgical, Mansfield, MA). "Second-look" arthroscopy was performed at an average of 9 months following the index surgical procedure. The reasons for this second surgery were recurrent instability in 7, pain in 6, and pain and stiffness in 2. In the 7 patients with recurrent instability, the Bankart repair was found to be completely healed in 3 (43%), partially healed in 1 (14%), and had recurred in 3 (43%); however, 6 of 7 were observed to have lax capsular tissue. In 4 of these cases, retrospective review of the index surgical procedure showed that a technical error had been made during the repair. Two cases had biopsy of the repair site on "second-look" at 6 to 8 months, and this showed residual polyglyconate polymer debris surrounded by a histiocytic infiltrate. In the remaining 8 cases with stable shoulders, the Bankart repair had completely healed in 5 cases (62.5%) and partially healed in 3 cases (37.5%). The higher failure rate with this approach compared with open approaches appears to result from improper patient selection and errors in surgical technique. There is some question concerning healing strength of the Bankart repair, although complete healing of the Bankart does not seem to be a prerequesite for shoulder stability. Success of the procedure might be expected to improve by selecting only patients with unidirectional, posttraumatic, anterior instability who are found to have a discrete Bankart lesion and well-developed ligamentous tissue.