Advanced Granulation Technology (AGT(TM)). An alternate format for serum-free, chemically-defined and protein-free cell culture media.

To overcome limitations of conventional milling technology, we investigated the application of fluid bed granulation for the production of dry-form nutrient media. Serum-free, protein-free and chemically-defined specialty media were produced in granulated format and compared with identical formulations manufactured by conventional methods. HPLC analysis of multiple lots of granulated materials demonstrated that biochemical constituents were precisely and homogeneously distributed throughout the granules and that nutrient levels were comparable to conventional formats. Comparison of medium performance in cell proliferation and biological production assays demonstrated equivalence with reference media. The fluid bed granulation process meets pharmaceutical quality requirements and may be applied to a broad range of nutrient formulations required for bioproduction applications.

Continuous, high capacity reconstitution of nutrient media from concentrated intermediates.

We designed an Integrated Media Preparation System (IMPS) for continuous, on-line preparation of cell culture media and delivery to intermediate storage vessels or directly to a bioreactor. Key components of the IMPS include: a high precision, continuous fluid mixing device; formulation-specific liquid medium concentrates; validated process controls and membrane filtration; and automated dispensing into large volume flexible plastic containers. The IMPS system is designed to produce sterile, single-strength liquid medium from common raw materials at a delivery rate of 1000-3000 liters per hour and will manufacture homogenous batches from several thousand liters to over 60,000 liters. Fortified nutrient media prepared from multi-component 50X concentrates have been demonstrated to accelerate bioreactor seed chains, increase product yield, and reduce the overall manufacturing cost of nutrient medium. A productivity matrix will analyze the fully-loaded costs and contrast alternative methods for media preparation against projected biological yield.

Total cholecystectomy utilization by diagnosis related group.

Utilization rates for total cholecystectomy by diagnosis related group in Sacramento, Calif, and Syracuse, NY, between 1981 and 1984 were studied. The two areas had similar hospital discharge rates and bed supplies. The data included 8989 discharges. Discharge rates for the combined cholecystectomy diagnosis related groups differed by only 1% to 3%. Syracuse mean stays for these categories exceeded those of Sacramento by 34% to 38% and were the principal cause of differences in hospital utilization for the procedure. These differences were related to more conservative community-wide physician practice patterns in Syracuse rather than to differences in the composition of the samples by age and payor status, or to the impact of length of stay variability. In the future, shorter stays for cholecystectomy, such as those of Sacramento, may become the norm for the entire nation.

Evidence that the mechanisms of fungitoxicity of 8-quinolinol and its bischelate with copper(II) are different.

The copper(II) contents of the growth media, Sabouraud dextrose and Czapek-Dox broths, and of the spore inocula of Aspergillus niger (ATCC 1004), Aspergillus oryzae (ATCC 1011), Trichoderma viride (ATCC 8678), and Myrothecium verrucaria (ATCC 9095) were determined by atomic absorption spectrophotometry in a graphite furnace. The test systems composed of Sabouraud dextrose broth and spore inocula of the four fungi contained only a little over 3% of the copper(II) required to form a minimal inhibitory concentration of bis(8-quinolinolato)copper(II). The test system of Czapek-Dox broth and A. oryzae contained slightly less than 65% of the copper(II) required to form a minimal inhibitory concentration of the bischelate of 8-quinolinol with copper(II). When the minimal inhibitory concentrations of 8-quinolinol and bis(8-quinolinolato)copper(II) were added simultaneously to the test system of Czapek-Dox broth and A. oryzae, 10% of the combined mixture of toxicants caused complete inhibition of growth indicating synergism between the toxicants. These results together with the observation that alpha-lipoic acid as well as small aliphatic thiol-containing compounds (cysteine, glutathione, dithioerythritol, and dithiothreitol) reversed the toxicity of 8-quinolinol but not the toxicity of bis(8-quinolinolato)copper(II) led to the conclusion that the mechanisms of fungitoxicity of both toxicants are different.

Syrian hamster lymphocyte cytogenetic technique and effects of blood storage on mitotic index.

The Syrian hamster has a diploid chromosome complement similar to humans in both number (2N = 44) and morphology. For comparative mutagenic studies with humans, a repeatable lymphocyte chromosome technique involving laboratory mammals is desirable. The reported hamster lymphocyte cytogenetic technique appears technically uncomplicated and repeatable while providing a sufficient number of metaphase cells for quantitative analysis. In order to determine the effects of storing whole-blood (in culture media) for varying time periods at 8 degrees C prior to adding a mitogen, the mitotic index was calculated following a 48-h culture period. Results indicated that storage up to six days can still result in a mitotic index adequate for cytogenetic analyses.