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Isolation of Staphylococcus species from the tonsils of healthy cattle and phage patterns of isolates.

Staphylococci were found in the tonsils of 121 (75.2%) of 161 cattle. There were 15 different species, 10 belonging to novobiocin-sensitive species. The most predominant species was S. simulans (79.3% of the 121 carriers), followed by S. aureus (20.7%), S. chromogenes (10.7%) and S. epidermidis (8.3%). The other 11 species were present in 0.8 to 5.8%. Twenty-six unidentifiable isolates were isolated from 26 (21.5%) carriers. Sixty-two (51.2%) of the 121 carriers yielded two to five Staphylococcus species together while only one species could be found in each of the other 59 (48.8%). Combinations of S. simulans and other species were most frequently encountered in 50 (41.3%) of the 121 carriers. Twenty-four (96.0%) out of 25 S. aureus isolates, 3 (42.9%) of 7 S. hyicus isolates and 45 (25.4%) of 177 coagulase-negative staphylococci (13 species and unidentifiable isolates) isolates were phage typable. Most of S. aureus isolates were lysed by bovine phages 119 (n = 16) or 116 (n = 5). Thirty-three (25.4%) of 45 coagulase-negative staphylococci typable isolates with Pulverer's phage set showed the phage pattern ph5/ph9/ph10/ph12/ph13/U4/U14/U16/++ +U20/U46. The tonsils of cattle thus appear to be a suitable environment for Staphylococcus species, particularly novobiocin-sensitive species.

Animals↗

Optimal boiler control through real-time monitoring of unburned carbon in fly ash by laser-induced breakdown spectroscopy.

A laser-induced breakdown spectroscopy (LIBS) technique has been applied for detection of unburned carbon in fly ash, and an automated LIBS unit has been developed and applied in a 1000-MW pulverized-coal-fired power plant for real-time measurement, specifically of unburned carbon in fly ash. Good agreement was found between measurement results from the LIBS method and those from the conventional method (Japanese Industrial Standard 8815), with a standard deviation of 0.27%. This result confirms that the measurement of unburned carbon in fly ash by use of LIBS is sufficiently accurate for boiler control. Measurements taken by this apparatus were also integrated into a boiler-control system with the objective of achieving optimal and stable combustion. By control of the rotating speed of a mill rotary separator relative to measured unburned-carbon content, it has been demonstrated that boiler control is possible in an optimized manner by use of the value of the unburned-carbon content of fly ash.

Journal Article↗

Oral vs intravenous dipyridamole echocardiography for detecting coronary artery disease.

The usefulness of the intravenous dipyridamole-echocardiography test (12-lead and two-dimensional [2-D] echo monitoring during dipyridamole infusion) in the diagnosis of coronary artery disease recently has been suggested. However, the intravenous form of dipyridamole is not available for clinical use in some countries and therefore the administration of oral dipyridamole has been employed in combination with echocardiography. In order to evaluate the relative usefulness of the oral (300 mg of pulverized tablets) vs the intravenous (up to 0.84 mg/kg in 10 min) dipyridamole-echocardiography test, we performed the two tests, on different days and in random order, in 28 inhospital patients: 21 had coronary artery disease (seven had one-vessel disease, eight had two-vessel disease, and six had three-vessel disease); seven patients had no significant coronary artery disease. For both tests, the diagnostic end-point was the development of a transient dyssynergy of contraction. Sensitivity was 95 percent for the intravenous and 52 percent for the oral dipyridamole-echocardiography test (p < 0.01); in positive cases, the dyssynergy after the dipyridamole administration appeared at 6.5 +/- 2.5 min for the intravenous and at 27.8 +/- 12.4 min for the oral test (p < 0.01). Specificity was 100 percent for both the intravenous and oral dipyridamole-echocardiography test. One or more extracardiac side effects (headache, gastrointestinal upset, flushing, etc) occurred in 61 percent of the intravenous and 68 percent of the oral tests (p = ns). Nine patients with a positive intravenous and oral dipyridamole-echocardiography test also had a positive exercise-electrocardiography test. A significant correlation between exercise time (ie, the time from onset of exercise and 0.1 m V of ST segment shift) and dipyridamole time (ie, the time from onset of dipyridamole administration and the development of frank dyssynergy) was present for the intravenous (r = 0.6, p < 0.05) but not for the oral test. We conclude that the oral dipyridamole-echocardiography test, in comparison with the intravenous dipyridamole-echocardiography test, has a lower sensitivity and requires a substantially longer imaging time. The dipyridamole time is related to exercise time for intravenous but not for the oral dipyridamole-echocardiography test.

Administration, Oral↗

Molecular and biochemical assays of cartilage components.

The procedure described below is useful for extracting proteins, nucleic acids, and glycosaminoglycans from 5-40 mg of cartilage or tissue-engineered cartilage samples. This extraction method will generate samples compatible with Western blot, RNase protection, dimethyl methylene blue (DMB) assay for glycosaminoglycan, Hoechst DNA assay, and hydroxyproline assay. Most soluble matrix molecules can be extracted from pulverized samples using 4 M guanidine HCl, during a 30-min period of vortex agitation at 4 degrees C. Shorter agitation times can give inadequate solubilization. The guanidine HCl-insoluble pellet must be re-extracted with guanidine thiocyanate buffer, to solubilize RNA additionally. The final insoluble pellet can be rinsed with ethanol and digested with papain, to quantify collagen content as well as other insoluble or crosslinked material. Samples between 1 and 5 mg may be directly digested with a small volume of papain buffer for DMB, hydroxyproline, and Hoechst DNA assays.

Adult↗

Isolation of DNA from tumor samples.

A method for producing high-molecular-weight DNA from pulverized tissue, nuclear fractions, or cultured cells. This isolation method relies on the powerful proteolytic activity of proteinase K combined with the denaturing ability of the ionic detergent sodium dodecyl sulfate. Ethylenediaminetetraacetic acid is included in the lysis buffer to inhibit DNases.

DNA, Neoplasm↗

Pseudomalabsorption of levothyroxine: a case report.

A 51-year-old woman who had been treated with levothyroxine sodium because of hypothyroidism after total thyroidectomy for thyroidal cancer was admitted to our hospital for persistent hypothyroidism despite large dose administration of levothyroxine (600 microg/day). The patient complained of severe general fatigue and body weight gain. Free thyroxine, free triiodothyronine and thyrotropin levels were 0.97 ng/dl, 1.55 pg/ml and 24.51 microU/ml, respectively, under oral administration of levothyroxine. Levothyroxine loading test performed by liquid form, pulverized tablets via nasogastric tube and intravenous administration revealed no evidence of malabsorption or metabolic disorder of levothyroxine, although oral intake of tablets was ineffective due to her factitiousness. We report here a possible case of "pseudomalabsorption of levothyroxine" to emphasize the clinical recognition of this disorder in patients with resistant hypothyroidism.

Dose-Response Relationship, Drug↗

A new mechanical device for declotting of hemodialysis access grafts: first clinical experience.

PURPOSE: To review the early clinical experience with a new mechanical thrombectomy device for declotting polytetrafluoroethylene (PTFE) hemodialysis access (HDA) grafts. METHODS: The XTD is a 6-F introducer-compatible device that pulverizes clot with a rotating curved tip driven by a flexible spiral shaft. Fragments are aspirated into a collection container. Fifty patients (32 women; mean age 66 years) with clotted 6-mm polytetrafluoroethylene HDA grafts were treated at 2 medical centers in a study spanning 22 months. In all, 59 procedures were performed on an outpatient basis under monitored conscious sedation. Ancillary techniques were used as needed to treat the underlying cause of the clot. Technical success was defined on a per-procedure basis as removal of sufficient thrombus to visualize the underlying disease without major device-related complications. RESULTS: Technical success was 100%; no device-related adverse events occurred. Immediate clinical success was achieved in 47/59 (80%) cases. Continued clinical success in survivors was 52% (30/58) at 1 month and 46% (26/56) at 3 months. Eliminating from analysis a high-risk patient subgroup (reocclusions, chronically clotted or unsalvageable grafts, untreatable central venous stenosis, and venous anastomoses resistant to wire/catheter passage) increased the 3-month clinical success to 59% (22/37). CONCLUSIONS: The XTD is a promising device capable of safely and effectively declotting PTFE grafts. Further investigation is warranted.

Aged↗

Cavitation bioeffects.

Acoustic cavitation takes place when tiny gas bubbles oscillate, grow, and collapse in liquid under the influence of ultrasonic field. This study reviews cavitation bioeffects that are found both in vivo and in vitro when exposed to either low- or high-power acoustics. Proposed mechanisms are discussed here as well based on theoretical studies, simulations and test bench experiments. Bioeffects are induced in living tissue once the gas bubble is, for instance, within a blood vessel in close vicinity to the endothelium or to the red blood cells. Conditions for inducing various bioeffects are discussed - from severe damage, such as cell necrosis, to delicate alterations, such as increased permeability of cell membrane. Present and potential applications for therapeutic purpose from stone pulverization and tissue ablation to gene transfection and transdermal delivery are reviewed including the growing use of artificial microbubbles.

Acoustics↗

Clinical correlations of aggrecan in the resected medial rectus muscle of patients with intermittent exotropia.

The purpose of this study was to elucidate the role of extracellular matrix components such as aggrecan, fibronectin, and laminin in the extraocular muscle of patients with strabismus. Resected tissues of the medial rectus muscle of 47 patients with intermittent exotropia obtained during recession-resection surgery were frozen under liquid nitrogen and pulverized by a Freezer/Mill to solubilize the tissue for enzyme immunoassay. The total amounts of aggrecan, fibronectin, and laminin in the resected tissue were correlated with clinical data of patients such as age, exodeviation, and refractive error. The amount of aggrecan decreased significantly with the advance of age (P < 0.0001, Spearman rank correlation test), while the amount of laminin or fibronectin had no correlation with age. Patients with basic type intermittent exotropia showed larger, although not significantly, amounts of aggrecan than those with convergence insufficiency type (P = 0.0538, Mann-Whitney U-test). The amount of aggrecan may be related to motor aspects of intermittent exotropia.

Adolescent↗

Aging reduces the bioavailability of even a weakly sorbed pesticide (carbaryl) in soil.

We investigated bioavailability and biodegradation of carbaryl (1-naphthyl methylcarbamate) in a soil with a long history of pesticide contamination from a storage facility located at Mamoon Kanjan, Pakistan. Carbaryl is weakly sorbed and generally considered to be easily degradable in soil. Extraction studies revealed that 49% of the total carbaryl in soil (88.0 mg kg(-1)) was not water-extractable and also not bioavailable, as demonstrated by inoculation of the contaminated soil with a carbaryl-degrading, mixed bacterial culture. Inoculation of the contaminated soil with the carbaryl-degrading culture showed that the bacteria were capable of degrading only the available (i.e., water-extractable) fraction of the pesticide. When the soil was pulverized in a ball mill to enhance the release of residue, an additional 19% of the carbaryl became bioavailable. A significant proportion of residue (approximately 33%) remained unavailable. The long (>12 years) contact time between the pesticide and soil (i.e., aging), allowing possible sequestration into soil nanopores and the organic matter matrices, is suggested to have rendered the pesticide unavailable for microbial degradation. High concentration (88.0 mg kg(-1)) in soil facilitated its persistence and sequestration. Results from the present study demonstrate that even a weakly sorbed and easily degradable pesticide, carbaryl, is effectively sequestrated in soil with time, rendering it partly inaccessible to microorganisms and affecting the bioavailability of the compound.

Adsorption↗

Volcanic activity: a review for health professionals.

Volcanoes erupt magma (molten rock containing variable amounts of solid crystals, dissolved volatiles, and gas bubbles) along with pulverized pre-existing rock (ripped from the walls of the vent and conduit). The resulting volcanic rocks vary in their physical and chemical characteristics, e.g., degree of fragmentation, sizes and shapes of fragments, minerals present, ratio of crystals to glass, and major and trace elements composition. Variability in the properties of magma, and in the relative roles of magmatic volatiles and groundwater in driving an eruption, determine to a great extent the type of an eruption; variability in the type of an eruption in turn influences the physical characteristics and distribution of the eruption products. The principal volcanic hazards are: ash and larger fragments that rain down from an explosion cloud (airfall tephra and ballistic fragments); flows of hot ash, blocks, and gases down the slopes of a volcano (pyroclastic flows); "mudflows" (debris flows); lava flows; and concentrations of volcanic gases in topographic depressions. Progress in volcanology is bringing improved long- and short-range forecasts of volcanic activity, and thus more options for mitigation of hazards. Collaboration between health professionals and volcanologists helps to mitigate health hazards of volcanic activity.

Disaster Planning↗

Time course observation of thyroid proliferative lesions and serum levels of related hormones in rats treated with kojic acid after DHPN initiation.

Time course changes in thyroid proliferative lesions as well as related hormone levels in the blood of male F344 rats given N-bis(2-hydroxypropyl)nitrosamine (DHPN: 2800 mg/kg body weight, single s.c. injection) as an initiation treatment followed by pulverized basal diet containing 0% (Group 2), 2% (Group 3) or 4% (Group 4) kojic acid (KA) were examined at Weeks 1, 2, 4, 8 and 12. As an untreated control group (Group 1), rats were given basal diet for 13 weeks and examined in the same manner. Serum T3/T4 levels in the DHPN + 2% KA and DHPN + 4% KA groups were significantly reduced as compared with the DHPN-alone group at each time point. Serum TSH levels in both DHPN + KA groups were significantly increased at each time point in a treatment period-dependent manner from Weeks 1 to 12, and the extent of elevation was more remarkable in the DHPN + 4% KA group. At Week 2, there were no statistically significant intergroup differences in liver T4-UDP-GT activities on a milligram microsomal protein basis. Histopathologically, no thyroid proliferative lesions were observed in the untreated control group or the DHPN-alone group. However, diffuse follicular cell hypertrophy and decreased colloid in the thyroid were apparent in all rats of the DHPN + KA groups at each time point. In addition, focal follicular cell hyperplasias and adenomas of the thyroid were observed at high incidence in the DHPN + 2% KA group from Week 4 and in the DHPN + 4% KA group from Week 8. Multiplicities of focal follicular cell hyperplasias and adenomas of the thyroid in the DHPN + 2% KA group were significantly greater than those in the DHPN + 4% KA group at Week 8. In the pituitary, an increase in the number of TSH producing cells with expanded cytoplasm was apparent from Weeks 4 to 12 in both DHPN + KA groups. These results strongly suggest that thyroid proliferative lesions were induced by KA administration due to continuous serum TSH stimulation through the negative feedback mechanism of the pituitary-thyroid axis, resulting from depression of serum T3 and T4.

Adenoma↗

Dose-threshold for thyroid tumor-promoting effects of orally administered kojic acid in rats after initiation with N-bis(2-hydroxypropyl) nitrosamine.

In order to evaluate the threshold dose of thyroid tumor-promoting effects of KA, male F344 rats were initiated with N-bis(2-hydroxypropyl) nitrosamine (DHPN; 2000 mg/kg body wt., single s.c. injection) and, starting 1 week later, received pulverized basal diet containing 0%, 0.002%, 0.008%, 0.03%, 0.125%, 0.5% or 2%KA for 20 weeks. Five rats each in the 0%, 0.125%, 0.5% and 2%KA groups were sacrificed at week 12, and 10 rats each in all groups at week 20. As an additional experiment, three groups without DHPN initiation received basal diet, a diet containing 0.5% or 2%KA for 20 weeks. The serum T4 levels were significantly decreased in the DHPN-initiated groups given 0.125%KA or more at week 12. No significant decreases in serum T3 levels were observed in the groups treated with DHPN + KA and a significant increase was evident in the 2%KA-alone group at week 20. Some rats in the DHPN + 2%KA group at weeks 12 and 20 and the 2%KA-alone group at week 20 showed pronounced elevation of serum TSH. Thyroid weights were significantly increased in the DHPN-initiated groups receiving 0.5% and 2%KA at weeks 12 and 20 and in the 2%KA-alone group at week 20. Histopathologically, the incidences of focal thyroid follicular cell hyperplasias in the DHPN-initiated groups treated with 0.125%, 0.5% and 2%KA at week 20 were 5/10, 10/10 and 8/8 rats, respectively. At week 20, adenomas were observed in 7/10 rats in the DHPN + 0.5%KA group and 8/8 rats in the DHPN + 2%KA group, and carcinomas were observed in 6/8 rats in the DHPN + 2%KA group. In the groups without DHPN initiation, only focal follicular cell hyperplasia was observed in 1/9 rats in the 2%KA-alone group. These results suggest that the no-observed-adverse effect for the thyroid tumor-promoting effect of KA is 0.03% (15.5 mg/kg/day) under the present experimental conditions, and that KA possesses weak tumorigenic activity in rats due to continuous serum TSH stimulation by a non-genotoxic mechanism.

Animals↗

Biopharmaceutical powders: particle formation and formulation considerations.

It is well known that protein/peptide-based drug formulations are more stable in the solid state than in the liquid state, thereby offering stability advantages in ambient temperature storage, product shipping/distribution, and long-term shelf life. Novel powder-based drug delivery systems recently emerging for applications in sustained release, inhalation, intradermal delivery, etc, add more value to protein solid dosage forms. Despite great research interests in understanding the drying effects on protein stability and a large collection of publications focusing on this area, systematic accounts of powder formation techniques are lacking. This review is to summarize a number of methods currently available for protein powder preparation. Some are common methods such as lyophilization, spray drying, pulverization, and precipitation, and some methods are more recently developed such as supercritical fluid precipitation, spray-freeze drying, fluidized-bed spray coating and emulsion precipitation. In addition to examining the individual process effect on protein stability that is always the focus of formulation scientists, this review also likes to evaluate each method from a more practical sense in terms of process versatility and scalability. The conclusion is that each method has its own advantages and the use of a method is formulation and application specific. With the understanding of the principles and advantages of these methods, it can benefit our choice on selecting appropriate techniques for preparing a desired protein powder formulation for specific applications.

Biopharmaceutics↗

Factors affecting disappearance of feedstuffs from bags suspended in the rumen.

Four experiments were conducted to examine the effect of various factors on in situ digestion of feedstuffs in the rumen of Holstein cows. In Exp. 1, the effect of various days, animals and animal diets on in situ digestion was evaluated. Measured disappearance of dry matter (DM) and nitrogen (N) from soybean meal (SBM) suspended in dacron bags was lowest when the host animal was fed a high grain diet. No effect due to animals, day of incubation or period of experimentation was observed. The effect of bag porosity and substrate particle size on DM and N disappearance from bags was studied in Exp. 2. Rip-stop nylon, a material with small pores, appeared to decrease the influx of digesting agents into bags and limited efflux of digested residues from bags when compared with bags made from dacron polyester material. Disappearance of DM and N from SBM and distillers grains (DG) in rip-stop nylon bags was greater when these feedstuffs were in the pulverized form rather than in the commercially processed form. In Exp. 3, dacron and Acropore bags, with pore sizes of 52 and 5 microns, respectively, were used to examine DM and N disappearance, in the rumen of cows, from total mixed diets containing various dietary N sources. Lower DM and N disappearance from Acropore bags was observed for each of the diets regardless of time spent in the rumen; much of the difference was established within the first hour. The effect of formaldehyde treatment on degradation of amino acids from SBM suspended in dacron bags in the rumen for 16 h was studied in Exp. 4. Increased resistance of SBM amino acids to degradation in the rumen was observed as a result of formaldehyde treatment levels as low as .3% by weight. A possible increase in protection of lysine from degradation relative to other amino acids was also observed.

Amino Acids↗

Elevation of a specific mRNA in longissimus muscle of steers fed ractopamine.

To address the hypothesis that some classes of growth promoters stimulate muscle protein synthesis in growing cattle, 23 crossbred steers were fed diets containing the phenethanolamine growth promoter ractopamine in a 140-d feeding trial. Steers received either no ractopamine, .18 or .36 mg.kg BW-1.d-1 ractopamine for 140 d or .36 or .72 mg.kg BW-1.d-1 ractopamine for 56 d. Longissimus muscle was obtained at slaughter and frozen in liquid N2. RNA was extracted by homogenization of pulverized frozen muscle in guanidinium isothiocyanate and centrifugation through cesium chloride. Polyadenylated mRNA was extracted by capture on oligo-dT columns. Ractopamine had no effect on total RNA or mRNA concentrations (P greater than .25). Hybridization of the RNA to a putative myosin light chain-1/3 (MLC-1/3) cDNA clone in a Northern blot indicated one heavy band (approximately 1 kb) with no evidence of extensive destruction of the RNA. A second, minor band (approximately 3 kb) also was observed in some samples. The MLC-1/3 cDNA clone was hybridized to 1- or 5-micrograms samples of total RNA, and the intensity of the resultant autoradiographs was quantified by laser densitometry. There was a statistical correlation between MLC-1/3 mRNA-micrograms RNA-1 and longissimus cross-sectional area (P less than .05) and average daily gain (P less than .025). The results suggest that ractopamine either increased the transcription of the putative MLC-1/3 gene and(or) increased the stability of MLC-1/3 mRNA in bovine longissimus muscle, either of which could result in an increase in specific myofibrillar protein synthesis.

Analysis of Variance↗

Pigment gallstone dissolution in vitro. Solubilization of brown bilirubinate and black polybilirubinate stone material by buffered solvents containing ethylenediaminetetraacetic acid, bile salts, and reducing thiols.

The efficacy of a buffered 1% ethylenediaminetetraacetic acid (EDTA)-2Na solution (pH 9.2) in solubilizing carefully pulverized material from brown bilirubinate and black polybilirubinate pigment stones can be intensified stepwise by admixtures of detergents and mucolytic active thiols. Solubilization effects were quantified either photometrically by measuring the dissolved calcium bilirubinate or gravimetrically by measuring the total weight loss of solids after a defined incubation period. Maximum effects were achieved when using a buffered solvent with 1 g/dl disodium-EDTA, 1 g/dl sodium taurocholate (NaTCA), and 2 g/dl N-acetylcysteine (NAC). Whereas admixtures of NAC enhanced the solubilization of brown bilirubinate stone material additionally by an average of 21.3% (related to the effect of an EDTA/NaTCA-containing solvent), black polybilirubinate material responded rather poorly and inconsistently to NAC (mean, 8.4 +/- 11.7%).

Bile Acids and Salts↗

Differential distribution of lumican and fibromodulin in tooth cementum.

The objectives of this study were to isolate and characterize the major proteoglycans of tooth cementum in relation to the tissue's mineralization. Cementum was collected from the root apex region of bovine molars and pulverized. It was first extracted with 6M guanidine-HCI, pH 7.4 (G-extract, mineral-unassociated) and then demineralized and extracted with 0.5M EDTA (E-extract, mineral-associated). Both extracts were applied to anion exchange and then molecular sieve chromatography to isolate proteoglycans. The fractions collected were assayed for chondroitin-(CS) and keratan sulfate (KS) containing proteoglycans using the monoclonal antibodies 2-B-6 and 5-D-4, respectively. It was found that the KS was the major glycosaminoglycan and was enriched in the G-extract fraction. The major KS fraction was then applied to 7.5% SDS-PAGE. The major broad band (69 kDa) was 5-D-4 positive in Western blot analysis and separated into two bands (46 kDa and 50 kDa) after treatment with keratanase II and endo-beta-galactosidase. These two proteins were transfered to PVDF membrane and analyzed for amino acid sequence. The results showed the major band (46 kDa) to be lumican and the minor (50 kDa) fibromodulin. In addition, based on the immunohistochemical study using a number of mono- and polyclonal antibodies including 5-D-4, anti-lumican core protein as well as anti-fibromodulin core protein antibodies, the KSPGs were found to be located almost exclusively in nonmineralized portions of cementum such as precementum and the pericementocyte area. These biochemical as well as immunohistochemical data suggest that the major KSPGs of cementum, lumican and fibromodulin, have a specific tissue distribution and may have regulatory roles in cementum mineralization.

Amino Acid Sequence↗