Tissues lyophilized

Activase (tissue Lyophile for IV injection Stable up to 8 hr at room temp, after lOOmg vial 3.5g L-Arg, lg... 

C. Hsu, H. Nguyen, D. Yeung, D. Brookes, G. Koe, T. Bewley, and R. Pearlman, Surface denaturation of solid-void interface—a possible pathway by which opalescent particulates form during the storage of lyophilized tissue-type plasminogen activator at high temperatures, Pharm. Res., 12, 69 (1995). 

Specimens for assay were prepared by suspending 500 mg of lyophilized brain in 50 ml of the buffer-papain mixture and hydrolyzing overnight at 37° C. The solution was then autoclaved for 10 minutes. This procedure yielded higher results than autolysis or prolonged enzymatic hydrolysis. After centrifuging off the debris, 0.5, 1.5, and 2.5 ml of the supernatant were used for assay. This represents 1, 3, and 5 mg of the original brain tissue per milliliter of final medium. 

Liver or brain tissue is homogenized and lyophilized. The dried tissue is suspended in water (1 mg/ml), incubated 3 days at 37°, and autoclaved 10 minutes to coagulate protein. After removing die coagulum, the supernatant is assayed at 1, 1.5, and 2.0 ml, which represents 0.4, 0.6, and 0.8 mg/ml of basal medium (weight/volume) of dried tissue. 

Liver tissue B12 is assayed by suspending 50 mg of lyophilized human liver in 100 ml of aconitate buffer to which 100 mg sodium metabisulfite is added. The suspension is autoclaved for 30 minutes at 16 psi to liberate the bound forms of the vitamin. Aliquots are diluted 1 10 after autoclaving to reduce metabisulfite toxicity on microorganism growth. Each milliliter now contains 50 pg of liver (w/v) 1.0, 1.5, and 2.0 ml of this hydrolyzate is added. After dilution to a volume of 5 ml, this represents an assay of 10, 15, and 20 ig of liver per milliliter of final solution. Normal liver contains between 2-14 mpg of vitamin B /mg liver powder. 

Aprotinin is a polypeptide consisting of a chain of 58 amino acid residues, which inhibits stoichiometrically the activity of several proteolytic enzymes such as chymotrypsin, kallikrein, plasmin, and trypsin. Aprotinin is obtained from bovine tissues and purified by a suitable process. It is stored as a bulk solution or lyophilized powder. The amount of two related substances des-Ala-des-Gly-aprotinin and des-Ala-aprotinin is determined by CZE with a 100% analysis. The relative migration times are 0.98 for des-Ala-des-Gly-aprotinin and 0.99 for des-Ala-aprotinin, and the specified limits are 8.0 and 7.5%, respectively. 

Over 50 different pyridazin-3-ones were evaluated for biological activity in a wheat (Triticum aestivum L.) test system described previously (1). Briefly, seeds were germinated in 9-cm petri dishes on three layers of filter paper. Pyridazinones were dissolved in acetone and the filter papers were impregnated with 1 ml of acetone solution. After the soluent evaporated, 10 ml of distilled water were added to form an inhibitor concentration of 100 yM. Seeds were planted directly on the moist papers and germinated for 4 days in a controlled environment chamber on a 16-hr photoperiod with 27+lC day temperature and 21+lC night temperature. Light intensity from both fluorescent and incandescent bulbs was 28 klux at dish level. Lipids were extracted and recovered from 1 g of lyophilized shoot tissue, separated into membrane and non-membrane lipids, and analyzed by gas chromatography as described (1). 

Activase (Alteplase) [CHO-expressed glycoprotein-recombinant human tissue plasminogen activator] Genentech Lyophilized (50 mg single-dose vial 100 mg single dose vial) 50 and 100 mg doses to be reconstituted with 50 or lOOmL of WFI, respectively Per 50 mg vial 1.7 g L-arginine 0.5 g phosphoric acid < 4 mg polysorbate 80 (under vacuum) Per 100 mg vial 3.5 g L-arginine 1 g phosphoric acid <11 mg polysorbate 80 (without vacuum) IV... 

Extraction of samples. Plant tissues were lyophilized after harvest and ground to a homogeneous powder in a Wiley mill (no. 20 mesh). Samples (0.500 g) of lyophilized whole leaf were extracted in 125 mL roundbottom flasks by steeping in 25 mL of chloroform for 30 min. The extract was filtered and the filtrate set aside. The extraction flask and filtered solids were rinsed with an additional 15 mL of fresh chloroform. The filtrate from the rinse was then combined with the original filtrate and the resulting solution was evaporated to dryness with a rotary evaporator. The dry solids were redissolved in a mixture of 20.0 mL methanol and 5.0 mL acetone using sonication to assist in dissolving of all solid material. 

This nonextractable radioactivity was probably the result of covalent binding of the furazolidone intermediates to endogenous macromolecules. The bioavailability of these bound tissue residues from the above pig residue depletion study was determined by feeding rats lyophilized samples of liver and muscle tissues from animals sacrificed at 0 and 45 days after the last treatment (132). Results showed that the fraction of the bound residues bioavailable to rats was in the range 16-41%. The toxicological impact of these bioavailable bound residues has not been yet determined. 

Before the extraction procedure may commence, the sample must be prepared in such a way that it is in a condition for extraction of the analyte(s). For analyzing sulfonamide residues in liquid samples such as milk, a pretreatment dilution step with water prior to direct fluorometric detection may be required (207). Dilution of milk with aqueous buffer (208) or sodium chloride solution (209) prior to sample cleanup has also been reported. For the analysis of honey a simple dissolution of the sample in water (210, 211) or aqueous buffer (212) is generally required. Semisolid samples such as muscle, kidney, and liver, require, however, more intensive sample pretreatment. The analyte(s) must be exposed to extracting solvents to ensure maximum extraction. The most popular approach for tissue break-up is through use of a mincing and/or homogenizing apparatus. Lyophilization (freeze-drying) of swine kidney has been carried out prior to supercritical-fluid extraction of trimethoprim residues (213). 

Figure 15. First step in preparing biological samples. Shown here is the lyophilizer manifold for removing water from samples. The flasks shown are 1200 ml. in volume and contain muscle tissue...

Figure 16. Lyophilized muscle tissue being compressed into an aluminum can. Between 150 and 250 grams of tissue can be compacted for counting...

Freeze-Dried Samples. Solid Materials and Tissues. These are first cut into approximately 1-inch cubes, frozen on a Teflon cookie sheet in a freezer, and placed in 1200-ml. freeze-dry flasks to capacity. The flasks are attached to the freeze-dried (lyophilizer) manifold, the valves are opened to vacuum, and the flasks are evacuated. The water from the tissues is trapped on a condenser. The dry tissues (drying time about 2-3 days) are removed from the lyophilizer and compressed into thin-walled aluminum cans with a Carver Laboratory press fitted with a special die, at about 24,000 lb. pressure (total). From 150-250 grams of the dry material, representing 500-1000 grams of fresh tissue, can be packed into a single can. The cans are sealed with a hand sealer and set aside for counting. Samples can be removed from the cans at a later date for chemical analysis or beta-emitter analyses. 

Wash, hand-peel, chop and lyophilize each plant tissue material for 3-4 h at 25°C (e.g., soybean (Table 17.1), potato, banana, eggplant, sweet potato and artichoke (Table 17.2). Grind the dry tissue to obtain a fine powder and select the particle size using sieves of lower than 200 pm mesh. Store the dry powder in a desiccator at 25°C and use it as the enzymatic source of PPO or peroxidase in the preparation of biosensor. Determine enzymatic activity and total protein content as described in Procedure 22 (in CD accompanying this book). Prepare the carbon paste electrode with dry tissue as described in the same procedure. 

Aloe latex contains anthraquinones and is completely different from aloe gel, a colorless gelatin obtained from the central portion of the aloe leaf. The mucilaginous parenchymous tissue is excised from fresh leaves and immediately utilized for pharmaceutical preparations, or lyophilized and kept dry until use. During extraction of the gel, it is practically impossible to prevent contamination by the leaf latex as the leaves are cut. On the other hand, in intact leaves, anthraquinones may diffuse into gel from the bundle sheath cells. To reduce such contamination, the starting material must be from varieties of aloe with a reduced anthraquinone content. 

The collection of plant tissue is quite different from animal tissue collection. The discussion of collection of plant and animal tissue by Dessauer et al.2S is detailed and helpful. However, the recommendations for procedures unique to plant tissue collection are somewhat misleading and outdated, especially when tropical collections are involved. Plant tissue can now be collected and transported as either fresh tissue (leaves and/or shoot cuttings) or preserved tissue the latter either as cryopreserved tissue (liquid nitrogen or dry ice) or as dried tissue (air-dried, herbarium-dried, lyophilized, or chemically dried). Ambient-temperature liquid chemical preservation techniques (such as those routinely done for herbarium plant specimens in the tropics) so far have been ineffective in maintaining adequate yields of high-quality DNA.15 It should be stressed again that the manner of collecting plant tissue is dictated by several other factors what macromolecule (DNA, RNA, or isozymes) will be examined, what type of nucleic acid extraction method will be used (or, more impor-... 

Various methods of dehydrating plant tissue prior to transport and macromolecular extraction have met with some success with a limited sampling of plant taxa. These have included lyophilization,35,36 dehydra-... 

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