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PubMed2003

Effect of sample preparation, length of time, and sample size on quantification of total lipids from bovine liver.

The objective was to evaluate the effect of sample preparation (pulverization under liquid nitrogen, homogenization, or sonication), time length of sonication (0-60 s), shaking in chloroform/methanol solvent (0, 2, 4, or 12 h), incubation in chloroform (0 or 12 h), and drying of extracted lipids at 50 degrees C (2, 4, 6, or 24 h), and sample size (50-250 mg) on quantification of total lipids from bovine liver. Pulverization under liquid nitrogen yielded the lowest recovery. Sonication was least time-consuming for sample preparation. Precise estimates and the greatest recovery were obtained with 30 s of sonication, at least 2 h of shaking in chloroform/methanol solvent, 12 h of incubation in chloroform, and at least 6 h of drying. Sample sizes of at least 150 mg gave precise estimates. The results demonstrate that sample preparation, time length of different steps of the extraction procedure, and sample size affect quantification of total lipid from bovine liver.

Animals↗
PubMed2005

Effects of solvent polarity and acidity on the extraction efficiency of isoflavones from soybeans (Glycine max).

Soy isoflavones have been correlated with beneficial health effects. The predominant chemical forms of isoflavones present may affect their biological activities. Choosing the solvent system that can accurately quantify the amounts of individual isoflavones present in these products is paramount. Our objectives were to compare frequently used solvent systems and to evaluate the effects of polarity and acidity on the recovery of isoflavones from soybeans. Isoflavones were extracted from pulverized Manokin soybeans using six solvent systems, which are the combinations of three polarity levels (83% acetonitrile, 80% methanol, and 58% acetonitrile) and two acidity levels (nonacidified and acidified). The pulverized soybean was stirred for 2 h in each solvent system before filtration and concentration using rotary evaporation. The extract was resuspended in 16% acetonitrile and analyzed by high-performance liquid chromatography. Recoveries of pure standards were evaluated with all solvent systems. Solvents with a higher polarity extracted a significantly higher amount of total isoflavones. For individual isoflavones, 58% acetonitrile (highest polarity) extracted either the highest amounts or no less than other solvents, while 83% acetonitrile (lowest polarity) extracted either the lowest amounts or no more than other solvents except for the aglycone form. Acidification significantly reduced the recovery of the malonylglucoside form and the total isoflavones. The recovery study revealed that acidification favored the chemical transformations of isoflavones during the extraction. Among the six solvent systems examined, 58% acetonitrile aqueous solution without acid was the best for extraction of isoflavones from soybeans.

Chromatography, High Pressure Liquid↗
PubMed1999

Solid dispersion of poorly water-soluble drugs: early promises, subsequent problems, and recent breakthroughs.

Although there was a great interest in solid dispersion systems during the past four decades to increase dissolution rate and bioavailability of poorly water-soluble drugs, their commercial use has been very limited, primarily because of manufacturing difficulties and stability problems. Solid dispersions of drugs were generally produced by melt or solvent evaporation methods. The materials, which were usually semisolid and waxy in nature, were hardened by cooling to very low temperatures. They were then pulverized, sieved, mixed with relatively large amounts of excipients, and encapsulated into hard gelatin capsules or compressed into tablets. These operations were difficult to scale up for the manufacture of dosage forms. The situation has, however, been changing in recent years because of the availability of surface-active and self-emulsifying carriers and the development of technologies to encapsulate solid dispersions directly into hard gelatin capsules as melts. Solid plugs are formed inside the capsules when the melts are cooled to room temperature. Because of surface activity of carriers used, complete dissolution of drug from such solid dispersions can be obtained without the need for pulverization, sieving, mixing with excipients, etc. Equipment is available for large-scale manufacturing of such capsules. Some practical limitations of dosage form development might be the inadequate solubility of drugs in carriers and the instability of drugs and carriers at elevated temperatures necessary to manufacture capsules.

Diffusion↗
PubMed2000

Extracorporeal shock wave lithotripsy of gallstones revisited: current status and future promises.

The improvement and refinement of extracorporeal shock wave lithotripsy (ESWL) has made this non-invasive treatment modality not only more effective, but also applicable to a larger population of gallstone patients. It can be performed safely on an outpatient basis. Advances in lithotripsy technology have made it possible to fragment stones into very small, sand-like particles (pulverization), which clear the gall-bladder faster than large fragments. Recent studies provide evidence that adjuvant bile acids may not be necessary in most cases in which pulverization is achieved. Good gall-bladder emptying appears both to promote the clearance of gallstones after ESWL and to decrease their recurrence. Although generally found to be more expensive than surgery if bile acids are used, ESWL should be cost-effective, as bile acids may not be necessary in all patients. Elderly patients with radiolucent, solitary and less than 30 mm gallstones can particularly benefit from lithotripsy.

Ambulatory Care↗
PubMed2004

Microbial populations and activities in the rhizoplane of rock-weathering desert plants. II. Growth promotion of cactus seedlings.

Four bacterial species isolated from the rhizoplane of cacti growing in bare lava rocks were assessed for growth promotion of giant cardon cactus seedlings (Pachycereus pringlei). These bacteria fixed N(2), dissolved P, weathered extrusive igneous rock, marble, and limestone, and significantly mobilized useful minerals, such as P, K, Mg, Mn, Fe, Cu, and Zn in rock minerals. Cardon cactus seeds inoculated with these bacteria were able to sprout and grow normally without added nutrients for at least 12 months in pulverized extrusive igneous rock (ancient lava flows) mixed with perlite. Cacti that were not inoculated grew less vigorously and some died. The amount of useful minerals (P, K, Fe, Mg) for plant growth extracted from the pulverized lava, measured after cultivation of inoculated plants, was significant. This study shows that rhizoplane bacteria isolated from rock-growing cacti promote growth of a cactus species, and can help supply essential minerals for a prolonged period of time.

Cactaceae↗
PubMed1988

Removal of focal atheromatous lesions by angioscopically guided high-speed rotary atherectomy. Preliminary experimental observations.

A new high-speed rotary atherectomy device, inserted over a guide wire and directed with an angioscope, offers the potential of restoring patency of outflow vessels by "boring out" the atheromatous lesion of the orifices of runoff vessels. The device was tested on 68 cadaver arteries with atheromatous lesions involving the superficial femoral, popliteal, and tibial arteries. This was performed with either free segments or in situ with the device placed through a popliteal arteriotomy. The gross results of rotary atherectomy were assessed by angioscopy, angiography, or both. The luminal surfaces were studied with scanning electron microscopy and transverse sections of vessels were studied with light microscopy. The pulverized atheroma, in colloidal suspension, was analyzed for particle size by Coulter counter. The effect of a colloidal suspension of atheromatous particles on distal capillary circulation was measured in animal experiments. Obstructive lesions were successfully removed in 36 of 37 stenotic arteries (97%) and 18 of 31 completely occluded arteries (58%), an overall efficacy of 54 of 68 (79%). In successfully atherectomized arteries, angioscopy and angiography demonstrated a widely patent, smooth, polished surface. Light microscopy demonstrated removal of the diseased intima with maintenance of the outer media and adventitia. The pulverized atheroma particles were generally smaller than red blood cells and injection of the colloidal atheroma into canine femoral arteries failed to produce local tissue injury. We conclude that in the human cadaver this atherectomy device effectively enlarges and recanalizes obstructed superficial femoral, popliteal, and tibial arteries.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiography↗
PubMed2006

Optimal sampling of rat liver tissue for toxicogenomic studies.

Different degrees of a toxic response between and within the various lobes of the liver have been observed in rodents following treatment with acetaminophen. This study was designed to compare 2 sampling methods of the rat liver for gene-expression analysis. Ten male Fischer 344/N rats, 12-14 weeks of age, were treated with vehicle (0.5% aqueous ethyl cellulose) or acetaminophen (APAP, 1500 mg/kg) and sacrificed 24 hours following dose administration. Two representative sections were collected from the left liver lobe, stained with hematoxylin and eosin (H&E), and evaluated independently by 2 pathologists. The central core of the left lobe was cubed and frozen. Five random cubes were conserved, while the remaining left lobe core was pulverized. From each of the 10 animals, 2 random cubes and 2 samples from the homogeneous, pulverized samples were prepared for microarray analysis. Histopathologic evaluation revealed a variable response of centrilobular necrosis within the left lobe. Multiple methods used to analyze the microarray data indicated that sampling technique was not a major contributor to the variability observed in the gene expression data; however, only the powdered samples clustered for all animals, even those with disparate histopathologic results. Additionally, a powdered sample provided the advantages of a homogenous sample pool and the ability to use sample aliquots for other analyses to include proteomics, metabonomics, and other molecular techniques.

Acetaminophen↗
PubMed1980

Initiation of DNA synthesis in the prematurely condensed chromosomes of G1 cells.

The objective of this study was to investigate whether G1 cells could enter S phase after premature chromosome condensation resulting from fusion with mitotic cells. HeLa cell synchronized in early G1, mid-G1, late G1, and G2 and human diploid fibroblasts synchronized in G0 and G1 phases were separately fused by use of UV-inactivated Sendai virus with mitotic HeLa cells. After cell fusion and premature chromosome condensation, the fused cells were incubated in culture medium containing Colcemid (0.05 micrograms/ml) and [3H]thymidine ([3H]ThdR) (0.5 microCi/ml; sp act, 6.7 Ci/mM). At 0, 2, 4, and 6 h after fusion, cell samples were taken to determine the initation of DNA synthesis in the prematurely condensed chromosomes (PCC) on the basis of their morphology and labeling index. The results of this study indicate that PCC from G0, G1, and G2 cells reach the maximum degree of compaction or condensation at 2 h after PCC induction. In addition, the G1-PCC from normal and transformed cells initiated DNA synthesis, as indicated by their "pulverized" appearance and incorporation of [3H]ThdR. Further, the initiation of DNA synthesis in G1-PCC occurred significantly earlier than in the mononucleate G1 cells. Neither pulverization nor incorporation of label was observed in the PCC of G0 and G2 cells. These findings suggest that chromosome decondensation, although not controlling the timing of a cell's entry into S phase, is an important step for the initiation of DNA synthesis. These data also suggest that the entry of a S phase may be regulated by cell cycle phase-specific changes in the permeability of the nuclear envelope to the inducers of DNA synthesis present in the cytoplasm.

Cell Cycle↗
PubMed1956

The freezing point depression of mammalian tissues in relation to the question of osmotic activity of cell fluid.

The freezing point depression of freshly excised frozen tissues, pulverized in a hydraulic press or in a mortar, is greater than that of plasma. Even at 0 degrees C. the freezing point depression of such homogenates increases significantly with time. Dilution data indicate that such freezing point data are valid. The presence of intact cells has been shown in smears of tissues pulverized in a mortar, but not in smears of those crushed in a hydraulic press. The osmolarity of various diluent solutions affects the calculated osmotic activity of tissue homogenates presumably because of delayed diffusion between the diluent and cell fluid. With a hypertonic NaCl diluent, spuriously low values of tissue osmotic activity are found from calculations assuming instantaneous mixing between homogenates and diluents. The limitations of data from cryoscopic experiments and from tissue-swelling experiments are discussed in relation to the basic question of whether or not cell fluid is isotonic to extracellular fluid.

Animals↗
PubMed1989

Effects of texture of food on chewing patterns in the human subject.

To elucidate the effects of food character on chewing patterns in humans, the electromyographic activity (EMG) of the chewing muscles and also the jaw movements were recorded in twenty-nine young subjects during ordinary chewing of five different foods. The results obtained were as follows. (a) The harder food materials showed a higher amplitude of the masseter EMG than the softer ones. (b) Concerning the number of chewing strokes and the elapse of time until the last swallowing action, subjects could be divided into two groups. (i) In the first major group, the number of chewing strokes and chewing time until the last swallowing action increased following increase of hardness of the food. This suggests that chewing force and chewing movements may be strongly influenced by the texture of food, especially its hardness. Further, the degree of pulverization of eating materials appears to be the major factor in controlling the swallowing action. (ii) In the second minor group, the chewing strokes and the chewing time were less influenced by the hardness of food. Here, a certain number of chewing strokes could be stimulating the swallowing centre in the brain and so induce swallowing, regardless of the degree of pulverization of the food.

Adhesiveness↗
PubMed1992

A comparison of tablets with oral suspension formulation of dipyridamole in thallium myocardial imaging.

Dipyridamole stress thallium imaging has been widely employed to diagnose and assess the extent of coronary heart disease in patients who cannot exercise. When oral dipyridamole administration was used, a wide range of results for sensitivity, specificity, hemodynamic response and side effect profile has been reported. The authors hypothesized that the formulation used for oral administration of dipyridamole plays a major factor in this variability, and that the pulverized form of dipyridamole will achieve faster and more consistent response than the standard tablet form. The authors studied 13 consecutive patients who underwent thallium scintigraphy. Eight patients received dipyridamole pulverized and dissolved in a glycol/aqueous base diluent (group A), and five patients received the standard form of dipyridamole (group B). In group A, mean peak systolic blood pressure decreased from 142 +/- 31 (mean +/- standard deviation) to 109 +/- 30 (P = .05), and mean diastolic blood pressure decreased from 76 +/- 14 to 51 +/- 5. The mean heart rate changed from 78 +/- 26 to 80 +/- 10. In group B, baseline systolic blood pressure was 165 +/- 12 and decreased to 156 +/- 7 at 45 minutes and to 155 +/- 14 at 90 minutes. Heart rate increased from baseline of 69 +/- 9 to 75 +/- 8 at 45 minutes and to 76 +/- 11 at 90 minutes. At 45 minutes, the systolic blood pressure of the 8 group A patients dropped by 33 +/- 19 mm Hg, whereas group B's changed by 9 +/- 6 mm Hg (P less than .005).(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗
PubMed1986

A cryoprotection method that facilitates cutting frozen sections of whole monkey brains for histological and histochemical processing without freezing artifact.

Cutting frozen sections of large (greater than 60 cc) blocks of monkey brain using the conventional procedures of infiltration with 30% sucrose as a cryoprotectant before freezing with pulverized dry ice often produces unacceptable levels of freezing artifact (FA) caused by displacement of tissue by ice crystals. Experiments investigating FA utilized perfusion-fixed brains from 46 monkeys and spanned combinations of cryoprotectants (glycerol, sucrose), freezing methods (dry ice or -75 degrees C isopentane), and fixatives (10% formalin, Karnovsky's or Timm's). The effects were evaluated by rating of FA severity in frozen sections of whole monkey brains. Minor FA appears as enlarged capillaries, more serious FA as large vacuoles, and both first appear midway between the periphery and center of the block. Stronger fixatives increased the severity of freezing artifact. The best method for eliminating FA was graded infiltration with up to 20% glycerol and 2% DMSO (in buffer or fixative), followed by rapid freezing in -75 degrees C isopentane. Although using a glycerol-DMSO infiltration before conventional freezing with pulverized dry ice or using conventional sucrose infiltration before freezing in isopentane gave better results than sucrose infiltration and dry-ice freezing, only the combination of glycerol-DMSO infiltration and freezing in isopentane produced consistently excellent results and virtually eliminated freezing artifact. To determine the effect of freezing with dry ice or isopentane on the rate of cooling in large blocks of CNS tissue, thermocouples were embedded in an 80-cc block of albumin-gelatin and frozen with the two methods. The rate of cooling (-3.5 degrees C/min) was twice as fast using isopentane.

Animals↗
PubMed2006

Procedures for the quantitative protein determination of urokinase and its inhibitor, PAI-1, in human breast cancer tissue extracts by ELISA.

The determination of the protein content of urokinase-type plasminogen activator (uPA) and its inhibitor, PAI-1, in breast cancer tissue extracts is used clinically to identify patients at risk to experience disease recurrence (metastasis) or early death. The serine protease uPA, in concert with its inhibitor PAI-1, promotes tumor cell adhesion, migration, and proliferation, as well as extracellular matrix degradation and, thus, facilitates tumor cell invasion and metastasis. The various technical steps to recover uPA and PAI-1 protein from archived breast cancer tissues and to quantitatively determine uPA and PAI-1 protein content in tumor tissue extracts by enzyme-linked immunosorbent assay (ELISA) are described in detail. The technical steps involved require fresh-frozen breast cancer tissue, a dismembrator machine (ball mill) to pulverize the tissue in the frozen state, detergent (Triton X-100) containing Tris-buffered saline to extract uPA and PAI-1 from the pulverized breast cancer tissue, an ultracentrifuge to separate the detergent fraction from cellular debris, uPA and PAI-1 ELISA kits, protein determination reagents, and a 96-well spectrophotometer (ELISA reader) to assess uPA, PAI-1, and total protein in the detergent extract. The uPA/PAI-1 ELISAs and the protein determination format described are robust and highly sensitive. In addition to the macromethod of tissue disintegration, we present a simple but sensitive micro-extraction procedure using cryostat sections or core biopsies as the source of breast cancer tissue. Such a technique allows rapid and quantitative determination of uPA and PAI-1, even in small breast cancer specimens.

Animals↗
PubMed2003

Clastogenic effects of streptozotocin on human colon cancer cell lines with gene amplification.

We investigated the clastogenic effects of the methylating agent streptozotocin (STZ) on two human colon cancer cell lines. COLO320DM and COLO320HSR are cell lines derived from a human neuroendocrine colon carcinoma. STZ produced a dose-dependent increase in the frequency of chromosomal aberrations in both cell lines (p < 0.05) and induced fragmentation and/or pulverization of COLO320DM and COLO320HSR chromosomes. This effect was dose and time dependent. Severe chromosome damage was also found in cells that had progressed beyond the first metaphase, and a higher percentage of metaphases showing pulverized chromosomes were found in cells after the second than after the first division. This seems to indicate that STZ has a persistent and delayed clastogenic effect on COLO320DM and COLO320HSR cells. In addition, STZ produced a marked depression of the mitotic index in both cell lines. These results demonstrate that human colon cancer cell lines COLO320DM and COLO320HSR are highly sensitive to STZ, and suggest that this antibiotic has a good potential as a chemotherapeutic agent for colon cancer.

Antibiotics, Antineoplastic↗
PubMed2005

Experiment and mechanism investigation on advanced reburning for NO(x) reduction: influence of CO and temperature.

Pulverized coal reburning, ammonia injection and advanced reburning in a pilot scale drop tube furnace were investigated. Premix of petroleum gas, air and NH3 were burned in a porous gas burner to generate the needed flue gas. Four kinds of pulverized coal were fed as reburning fuel at constant rate of 1g/min. The coal reburning process parameters including 15% approximately 25% reburn heat input, temperature range from 1100 degrees C to 1400 degrees C and also the carbon in fly ash, coal fineness, reburn zone stoichiometric ratio, etc. were investigated. On the condition of 25% reburn heat input, maximum of 47% NO reduction with Yanzhou coal was obtained by pure coal reburning. Optimal temperature for reburning is about 1300 degrees C and fuel-rich stoichiometric ratio is essential; coal fineness can slightly enhance the reburning ability. The temperature window for ammonia injection is about 700 degrees C approximately 1100 degrees C. CO can improve the NH3 ability at lower temperature. During advanced reburning, 72.9% NO reduction was measured. To achieve more than 70% NO reduction, Selective Non-catalytic NO(x) Reduction (SNCR) should need NH3/NO stoichiometric ratio larger than 5, while advanced reburning only uses common dose of ammonia as in conventional SNCR technology. Mechanism study shows the oxidization of CO can improve the decomposition of H2O, which will rich the radical pools igniting the whole reactions at lower temperatures.

Air Pollutants↗
PubMed1991

Gallstone fragmentation during biliary lithotripsy: effect of stone composition and structure.

In vitro lithotripsy with the Siemens Lithostar was conducted on 36 radiolucent or minimally calcified gallstones housed in an anthropomorphic phantom. The ease and pattern of fragmentation were correlated with global composition for the entire stone, regional or microcomposition (determined by Fourier-transform infrared spectroscopy), and microstructure (determined by scanning electron microscopy). Stones made up of more than 62% cholesterol required 50% more shock waves to pulverize all fragments to 0.3 cm or less than did stones of less than 62% cholesterol (p less than .01). An inverse relationship was found between the number of shock waves needed for fragmentation and the cholesterol content (r = .77). Although a broad range of fragmentation responses occurred, little variation was seen in the ease of fragmentation within stone families. The majority of stones fractured along radially oriented cholesterol plates, but one third of stones treated showed initial chipping or flaking at the periphery before radial fracture. This type of peripheral erosion most often occurred in stones with peripheral pigment rims. These stones required more shock waves and lagged in pulverization compared with more homogeneous cholesterol stones. The efficiency of fragmentation during biliary lithotripsy correlates with the stones' global cholesterol content. A stone's architecture, as reflected by its regional composition and microstructure, partially predicts the mechanism of fragmentation. These in vitro data may be useful in further refining criteria for selecting patients and understanding the fragmentation process.

Bile Pigments↗
PubMed1997

Length-reduction method for man-made mineral fibers for biological experiments.

A simple fiber length-reduction method was developed to obtain a large amount of fiber samples with different length distributions for use in various biological experiments. This press method is only to press a raw fiber sample charged in a stainless cylinder at an adequate pressure, and is effective for man-made mineral fibers (MMMF) such as glass wool, rock wool and wool and refractory fibers (ceramic fibers, mullite fibers) and some brittle natural mineral fibers such as fibrous brucite and wollastonite. The mean fiber-length of man-made mineral fibers became shorter with the increase in the pressure applied without diameter change. We could obtain a length-reduced fiber sample with a suitable length distribution by this method. This press method is therefore a size-selective method able to produce a large amount of pulverized fiber sample depending on the press cylinder size for biological experiments. A very small amount of non-fibrous particles with aspect ratios (length vs. diameter) of under 3 was seen in the pulverized fiber samples. To eliminate such non-fibrous particles as well as too long fibers from the sample, separation by sedimentation in water was somewhat effective.

Aluminum Silicates↗
PubMed2001

Osteosynthesis and allogeneic bone grafting in complex osteoporotic fractures.

Thirty patients with osteoporotic fractures were treated operatively. An allogeneic bone transplant was used in combination with a conventional osteosynthesis in each patient. The bone graft was pulverized in a bone mill and used as a substitute graft to fill the bone defect, add stability, and enhance bone union. Fractures healed without complications in 20 patients. The osteosynthesis failed in 4 patients. The fracture failed to unite in an additional 3 patients. One deep infection occurred. A biopsy taken from the allogeneic bone at plate removal after fracture union demonstrated mature bone and new bone formation. The use of pulverized allograft bone for large bone defects in patients with osteoporotic fractures yields acceptable results with no adverse effects.

Aged↗