Production of primary cell cultures on a semi-industrial scale. 1. Modification of the trypsinization procedure--possible means to improve cell yield

To produce large quantities of primary cell suspensions on efficient technique is described which is named "Biomix-Method". By this technique first tissue mince is performed automatically using tissue amounts between 300 g and 500 g. Trypsinization takes place in a three-litre trypsinization vessel using a stirring mechanism according to a defined trypsinization mode. In regard to optimal cell seeding concentration and a justifiable productivity one kidney of newborn or suckling calf yields 6--9 1 cell suspension and that of piglet 1,8-2,3 1, respectively. Proliferation behaviour and effectivity of primary cells are discussed and compared with other cell systems.     

Association of hyponatremia and hyperglycemia with outcome in dogs with congestive heart failure

Objective: To evaluate plasma sodium and glucose concentrations in dogs with congestive heart failure (CHF) prior to treatment and evaluate the differences between survivors and non‐survivors. Design: Retrospective study. Animals: Fifty‐nine dogs with CHF prior to receiving cardiac medication. Interventions: None. Measurements and main results: The mean plasma sodium concentration in dogs with CHF was below the reference range (144–156 mmol/L) and significantly lower (P=0.009) in non‐survivors (141±6 mmol/L) compared with survivors (147±4 mmol/L). The mean plasma glucose concentration was above the reference range (76–117 mg/dL) and significantly higher (P=0.004) in non‐survivors (128±52 mg/dL) compared with survivors (100±13 mg/dL). Forty‐four percent of non‐survivors had concurrent low plasma sodium and high plasma glucose concentrations, whereas no survivors had both abnormalities (P<0.0001). Conclusions: Lower plasma sodium and higher plasma glucose are associated with a worse outcome in dogs with CHF.     

Induction of phase II detoxification enzymes in rats by plant-derived isothiocyanates: comparison of allyl isothiocyanate with sulforaphane and related compounds

Plants of the family Brassicaceae contain high levels of glucosinolates. The latter compounds are degraded to isothiocyanates, some of which have been shown to be potent inducers of phase II detoxification enzymes in vitro. In the present study, the ability of six plant-derived isothiocyanates (allyl isothiocyanate, iberverin, erucin, sulforaphane, iberin, and cheirolin) to increase tissue levels of the phase II detoxification enzymes quinone reductase (QR) and glutathione S-transferase (GST) in a variety of rat tissues has been compared. At the low dose level employed (40 micromol/kg/day), cheirolin was without effect in any tissue. All of the other isothiocyanates, however, increased GST and QR activities in the duodenum, forestomach, and/or the urinary bladder of the animals, with the greatest effects being seen in the urinary bladder. With the exception of cheirolin, little difference was observed in the inductive activity of the various isothiocyanates. Phase II enzymes are known to protect against chemical carcinogenesis, and the selectivity of isothiocyanates in inducing such enzymes in the bladder is of interest in view of recent epidemiological studies showing a decreased incidence of cancer of this organ in individuals with a high dietary intake of Brassica vegetables.     

A high-resolution radiation hybrid map of the human genome draft sequence

We have constructed a physical map of the human genome by using a panel of 90 whole-genome radiation hybrids (the TNG panel) in conjunction with 40,322 sequence-tagged sites (STSs) derived from random genomic sequences as well as expressed sequences. Of 36,678 STSs on the TNG radiation hybrid map, only 3604 (9.8%) were absent from the unassembled draft sequence of the human genome. Of 20,030 STSs ordered on the TNG map as well as the assembled human genome draft sequence and the Celera assembled human genome sequence, 36% of the STSs had a discrepant order between the working draft sequence and the Celera sequence. The TNG map order was identical to one of the two sequence orders in 60% of these discrepant cases.     

Escherichia coli challenge and one type of smectite alter intestinal barrier of pigs

An experiment was conducted to determine how an E. coil challenge and dietary clays affect the intestinal barrier of pigs. Two groups of 32 pigs （initial BW： 6.9 ± 1.0 kg） were distributed in a 2 x 4 factorial arrangement of a randomized complete block design （2 challenge treatments： sham or E. coil, and 4 dietary treatments： control, 0.3% smectite A, 0.3% smectite B and 0.3% zeolite）, with 8 replicates total. Diarrhea score, growth performance, goblet cell size and number, bacterial translocation from intestinal lumen to lymph nodes, intestinal morphology, and relative amounts of sulfo and sialo mucins were measured. The E. coli challenge reduced performance, increased goblet cell size and number in the ileum, increased bacterial translocation from the intestinal lumen to the lymph nodes, and increased ileal crypt depth. One of the clays （smectite A） tended to increase goblet cell size in ileum, which may indicate enhanced protection. In conclusion, E. coli infection degrades intestinal barrier integrity but smectite A may enhance it.     

Microbial diffusion assay for antibiotics in feeds using a simplified design

Results are compared for the microbiological analysis of antibiotics in feeds using the AOAC plate diffusion assay and the simplified 2-plate assay. Five antibiotics, bacitracin, chlortetracycline, oxytetracycline, penicillin, and streptomycin, were used to supplement feed extracts at levels of 100 and 25 micrograms antibiotic/g feed (bacitracin at 100 micrograms/g only). For bacitracin at the one level and for penicillin at both levels, the 2-plate design yielded significantly more accurate results than those of the AOAC assay. The same was true for the 25 micrograms/g level of oxytetracycline and the 100 micrograms/g level of streptomycin. For streptomycin at the 25 micrograms/g supplementation, the AOAC assay results showed better accuracy. There was no significant difference in results between the 2 designs for oxytetracycline at 100 micrograms/g and chlortetracycline at either level. The accuracy and precision of the results for the 2-plate design are equivalent to or better than those obtained using the AOAC design; in addition, the 2-plate assay is less labor-intensive, is more cost-effective, and is able to determine reasonable conditions of similarity.     

Chemistry and world food supplies

Much of the unprecedented increase in developing countries' food production in the past two decades is due to chemical-based technologies and to the use of agricultural chemicals. However, these successes were won under generally favorable conditions of soil, climate, and irrigation water availability. The challenge of the future is to broaden the base of increased food production to include areas less well endowed with natural and economic resources. Chemistry and chemicals must play vital roles in this venture. Innovative chemical and biochemical approaches must be called upon to produce crop varieties, animal strains, and associated technologies to overcome constraints such as insects and diseases, acid and alkaline soils, and drought conditions. Genetic engineering will probably be a primary mechanism to achieve this goal.