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Lymphoproliferative diseases of fowl: chromosome breaks caused in lymphocytes by JM-V herpesvirus.

Chromosome preparations were made of bone marrow cells and peripheral lymphocytes isolated from chicks that developed leukemia following infection with JM-V herpes-virus. Karyotypic analysis revealed a high frequency of chromosome breaks and aneuploidy, as well as some pulverization of chromosomes. The number of chromosome breaks began to increase at 2-3 days post infection, and by 5 days post infection it reached 12.7% of bone marrow cells and 17.2% of peripheral lymphocytes. Similarly, the number of aneuploid metaphase figures increased rapidly and reached 12% of bone marrow cells and 19% of peripheral lymphocytes at 5 days post infection. Some specificity was observed in the chromosomes that were affected.

Aneuploidy↗

Mimosine is a potent clastogen in primary and transformed hamster fibroblasts but not in primary or transformed human lymphocytes.

The cytogenetic effects of mimosine, a naturally occurring plant amino acid known to arrest cell-cycle progression at the G1-S border in cultured cells, have been studied. It was found that mimosine inhibits the cell-cycle progression in a dose-dependent manner in primary and transformed Chinese hamster fibroblasts as well as primary lymphocytes and transformed lymphoblastoid cells of human origin. In the Chinese hamster fibroblast cells, the first division metaphases analysed were found to be highly damaged or pulverized. The damaged cells which could pass through the next cell division, showed very high frequencies of sister chromatid exchanges (SCEs) compared with untreated second division cells. No such cytogenetic alterations could be detected in the human cells. The absence of clastogenic effect in cells of lymphoid origin appears to be related to the known capacity of these cells to undergo apoptosis, thereby efficiently eliminating cells with high frequencies of chromosomal aberrations. Our study demonstrates the clastogenic potency of mimosine and suggests the need for a careful interpretation of the results while using mimosine for cellular or molecular studies pertaining to cell cycle events.

Animals↗

High pressure induces G2 arrest in murine erythroleukemia cells.

The effect of high pressure on proliferation and cell cycle progression was examined using murine erythroleukemia (MEL) cells. The MEL cells were exposed to high pressures (0.1-130 MPa) and then cultured for 5 days at atmospheric pressure. The proliferation of MEL cells was unaffected up to 60 MPa, but was suppressed at 80-110 MPa. Above 120 MPa, the cells were fragmented. The cell cycle analysis of 80 MPa-treated MEL cells showed that the cells in S phase are most sensitive to high pressure and they arrest in G2 phase. Interestingly, G2-arrested cells reinitiated DNA synthesis, resulting in giant cells with high DNA contents. Furthermore, when such G2-arrested cells were exposed to caffeine, premature mitosis, characterized by chromosome pulverization, was observed. These results suggest that the suppression of proliferation in high-pressure-treated MEL cells is associated with G2 arrest following S phase delay. Thus, it seems valuable to apply high pressure to the investigation of the cell cycle.

Adenine↗

Developmental changes in embryonic and extra-embryonic insulin-like growth factor-I tissue concentrations in the turkey embryo.

The ontogeny of insulin-like growth factor-I (IGF-I) embryonic and extra-embryonic tissue concentrations were determined in the developing turkey embryo. At 2-d intervals, starting on Day 6 of incubation, individual tissues (n = 8 for each stage of incubation) were removed, weighed, pulverized, and extracted in 1 M acetic acid for IGF-I determination. Amniotic and allantoic fluid were collected starting on Day 8, serum on Day 12, and analyzed for IGF-I levels. Serum IGF-I levels were the lowest on Days 12 and 28 of incubation (5.9 and 9.5 ng/mL), respectively, and the highest on Day 20 (16.2 ng/mL). Allantoic and amniotic fluid IGF-I concentrations were essentially unchanged during incubation. Extra-embryonic tissue IGF-I levels increased in both the yolk sac and chorioallantoic membranes as incubation advanced with concentrations being 8- to 10-fold greater in the chorioallantois. Embryo tissue IGF-I concentrations varied greatly with regard to tissue and stage of development. Brain IGF-I levels were the highest on Day 8 and lowest on Day 26 (423 vs 35 pg/mg protein, respectively). Tissue IGF-I pattern in the heart mirrored that of brain. Liver IGF-I was low (< 40 pg/mg protein) from Day 10 to 20 and undetectable on Days 22 to 28. Muscle IGF-I levels were similar in the final days of development to those observed in early incubation. Bone IGF-I levels were highest in mid-incubation and lowest on Day 26. Peptide levels in the gastrointestinal tract (GI) tract and gizzard were dissimilar in that IGF-I was elevated in the GI tract in early incubation and declined with advancing incubation, whereas gizzard IGF-I levels peaked in late incubation. It is apparent that tissue synthesis of IGF-I is differentially regulated within a given tissue and stage of incubation during embryogenesis in the turkey embryo.

Allantois↗

Inhibition of thyroid iodine uptake and organification in rats treated with kojic acid.

In order to elucidate the mechanisms of reduction of serum thyroid hormones caused by continuous administration of kojic acid (KA) and its thyroid tumor-promotion effects, male F344 rats were given pulverized basal diet containing 0.008%, 0.03%, 0.125%, 0.5%, or 2% KA for 4 weeks. As an untreated control group, additional rats were given basal diet alone for the same period. The thyroid 125I uptake was significantly decreased in the groups receiving 0.03% or more. In addition, significant reduction of organic formation of iodine and serum T3 and T4 levels were observed in the 2% KA group along with pronounced elevation of serum (TSH). Both absolute and relative thyroid weights were significantly increased in the groups receiving 0.5% of KA or more. Histopathologically, decreased colloid in the thyroid follicles and follicular cell hypertrophy in the thyroid were apparent at high incidences in the groups given 0.03% or more. In addition, thyroid capsular fibrosis was evident in all rats of the 2% KA group. In quantitative morphometrical analysis, the ratio of the area of follicular epithelial cells to the area of colloids was significantly increased in the groups given 0.03% KA or more. The results suggest that KA alteration of thyroid-related hormone levels in the 2% KA group are due to inhibition of iodide uptake and iodine organification in the thyroid, with tumor-promoting effects on development of thyroid proliferative lesions, probably secondary to prolonged serum TSH stimulation resulting from negative feedback through the pituitary-thyroid axis.

Animals↗

Photodeactivation of ethyl violet: a potential hazard of Sodasorb.

Breathing circuit cannisters containing functional CO2 absorbent are critical to prevent rebreathing CO2 during general anesthesia using closed or semiclosed breathing systems. Ethyl violet is the indicator dye added to Sodasorb to indicate impending exhaustion of the absorbent. A case of CO2 rebreathing due to failure of ethyl violet indicator in exhausted Sodasorb was encountered. Laboratory investigation demonstrated that dye failure could result from photodeactivation caused by fluorescent lights. Using a fixed intensity fluorescent light source and quantitative spectrophotometric analysis, a highly significant dose-response relationship was demonstrated between duration of light exposure and the decrease in ethyl violet concentration. After 24 h of fluorescent light exposure with a received flux density of 46 nwatts/cm2 at 254 nm, the concentration of functional ethyl violet remaining in pulverized Sodasorb was 16% of the baseline value. Furthermore, using multiple light sources of various intensities, the greater the intensity of light, the more rapid the rate of decline of the ethyl violet concentration. It is recommended to minimize the problem by using ultraviolet filters and incorporating additional ethyl violet in Sodasorb. Finally, ethyl violet undergoes temporal deactivation after a Sodasorb container is opened, even if it is stored in the dark.

Absorption↗

Characterization of thioether compounds formed from alkaline degradation products of enflurane.

BACKGROUND: Renal toxicity has occasionally been observed after enflurane anesthesia. Although originally attributed to its oxidative metabolism to inorganic fluoride, serum levels of inorganic fluoride appear to be small to explain these renal effects. Formation of potentially nephrotoxic halogenated alkenes during alkaline degradation in carbon dioxide absorbers and subsequent bioactivation via the glutathione conjugation pathway may be considered as an alternative mechanism for renal toxicity. The aim of this study was to characterize the thioethers formed chemically and biosynthetically. METHODS: Alkaline degradation of enflurane was achieved by stirring with pulverized potassium hydroxide. Volatile degradation products were analyzed by 19F nuclear magnetic resonance (NMR) analysis of head space gasses trapped in dimethyl sulfoxide (DMSO). Thioethers were generated chemically by trapping head space gasses in DMSO containing N-acetyl-L-cysteine or 2-mercaptoacetic acid as model thiol compounds. Glutathione conjugates were generated biosynthetically by passing head space through rat liver fractions in presence of glutathione. Products formed were analyzed by gas chromatography-mass spectroscopy and 19F-NMR. RESULTS: Direct analysis of head space gasses showed formation of 1-chloro-1,2-difluorovinyl difluoromethyl ether and two unidentified fluorine-containing products as alkaline degradation products of enflurane. When trapped in DMSO-N-acetyl-L-cysteine-triethylamine, N-acetyl-S-(2-chloro-1,2-difluoro-1-(difluoromethoxy)ethyl)-L-cysteine was identified as the major product. Another N-acetyl-L-cysteine S-conjugate formed was N-acetyl-S-(2-chloro-1,1,2-trifluoroethyl)-L-cysteine, a potent nephrotoxin in rats. 19F-NMR analysis of glutathione conjugates formed after incubation with rat liver fractions resulted in formation of corresponding S-conjugates. CONCLUSIONS: The current study demonstrates that alkaline degradation products of enflurane can be conjugated to thiol compounds, forming S-conjugates that could theoretically contribute to adverse renal effects observed occasionally with enflurane anesthesia. The N-acetyl-L-cysteine S-conjugates identified may be biomarkers to assess exposure of humans to alkaline degradation products of enflurane.

Acetylcysteine↗

The effects of particle size and moisture content on the emanation of Rn from coal ash.

Measurements of the emanation coefficients of 222Rn from coal ash were performed. Samples of coal ash from a stoker-fired furnace were mechanically sized into four categories. The mean 226Ra concentration of these categories ranged from 11.82-16.77 dpm g-1 and increased as a function of decreasing particle size. The mean bulk density and mean specific gravity of these categories ranged from 0.488-0.944 g cm-3 and 2.017-2.390 g cm-3, respectively, and both increased as a function of decreasing particle size. Samples of coal ash from four pulverized coal-fired furnaces were obtained, and demonstrated mean 226Ra concentrations ranging from 6.44-7.59 dpm g-1. The mean bulk density and mean specific gravity of these samples ranged from 1.254-1.520 g cm-3 and 2.357-2.588 g cm-3, respectively. Construction of chambers and methods for measuring 222Rn emanation coefficients are described. The measured emanation coefficient is shown to be insensitive to ingrowth time at times greater than three days. Measurements of emanation coefficients from ash samples were conducted at moisture contents of 0, 1.0, 10, 20 and 40% moisture by weight, and showed an increase in the emanation coefficient as a function of moisture content up to about 20% moisture by weight. Above 20% moisture content, most samples showed a trend toward decreasing emanation coefficients. The fractionated ash samples showed an increase in the emanation coefficient as a function of decreasing particle size. An experiment was also conducted which confirmed that sample depth was insignificant in explaining the observed emanation coefficient over the range of depths used in the other experiments.

Air Pollutants↗

Chernobyl fallout in southern and central Finland.

To study the levels and distributions of radionuclides released in the Chernobyl accident, we sampled surface peat from 62 sites in Southern and Central Finland and measured 131I, 134Cs, 137Cs, 132Te, 140Ba, 103Ru, 90Sr, 141Ce, and 95Zr. The distribution of fallout activities was highly uneven, depending on movement of the contaminated air mass and rainfall distribution during the critical days. The highest values observed were 420 kBq m-2 of 131I and 70 kBq m-2 of 137Cs. The nuclide ratios showed wide and partly unexpected variations. The high-boiling-point, or nonvolatile, elements Ce and Zr were spread mostly on a 200-km-wide zone extending across Finland from southwest to northeast. The more volatile elements, I, Ce, and Te, showed quite a different, more widespread, fallout distribution, while an intermediate behavior was observed for Ba, Ru, and possibly Sr. These results can be explained by assuming that pulverized nuclear fuel material released in the reactor explosion on 26 April reached Finland via Poland and the Baltic Sea and traversed the country along the above-mentioned narrow zone, while volatile material, evaporated in the reactor fire from 26 April to 5 May, arrived in several waves and was consequently more widely and evenly spread. From their elemental melting and boiling points, Ru and Mo would appear to belong to the nonvolatile group and Sr to the volatile. Yet, their actual behaviors were opposite; Ru in particular was found in the nonvolatile as well as the volatile fallout, possibly because Ru activities were present in the fuel partly in the metallic state and partly as volatile oxides.

Accidents↗

Rapid analytical technique to identify alpha emitting isotopes in water, air-filters, urine, and solid matrices using a Frisch Grid detector.

A 5-inch-diameter Frisch Grid gas-proportional ionization chamber was utilized at Brookhaven National Laboratory (BNL) to rapidly characterize and quantify alpha-emitting actinides in unprocessed water, soil, air-filter, urine, and solid matrices. Instrument calibrations for the various matrices were performed by spiking representative samples with National Institute of Standards and Technology traceable isotopes of 230Th, 232U, 236Pu, and 243Am. Detection efficiencies were typically 15-20% for solid matrices (soil, concrete, filters, dry urine) and 45% for mass-less water samples. Instrument background over a 512-channel alpha-energy range of 3-8 MeV is very low at 0.01 cps. At optimum efficiency, minimum detectable levels of 0.56 mBq Kg(-1), 74 mBq L(-1) and 14.8 mBq filter(-1) were achievable for 40 x 10(-6) Kg soil, 1 x 10(-3) L tap water (or urine), and 4.5 cm diameter air-filter samples, respectively, each counted for 60 min. Data and spectra are presented showing the quality of results obtained using untreated samples obtained from the BNL Graphite Research Reactor Decommissioning Project. These samples contained Bq to MBq per gram amounts of (239,240)Pu, 241Am, and/or (234,235/238)U (as well as other beta/gamma emitters). Data and spectra are also presented for a very finely pulverized and homogeneous U.S. DOE/RESL soil reference standard (spiked with 239Pu, 241Am, and 233U) that was used to assess precision, accuracy, and reproducibility. Although this technique has its limitations, the advantages are (1) minimal sample preparation, (2) no separation chemistry required, (3) no chemical or hazardous waste generated, and (4) ability to immediately characterize and quantify alpha-emitting nuclides in most matrices. The benefits of this technique to the BNL/DOE Project Managers were rapid (1-2 d) turn-around times coupled with significant cost savings, as compared to commercial off-site analyses.

Air Pollutants, Radioactive↗

Solid-state phosphorus-31 nuclear magnetic resonance differentiation of bone mineral and synthetic apatite used to fill bone defects.

Bioabsorption of synthetic apatite compounds used to promote bone healing and remodeling has been difficult to evaluate. In this study, solid-state phosphorus-31 nuclear magnetic resonance (NMR) has been used to characterize and quantitate bone mineral and a synthetic apatite in order to establish a model for bioabsorption studies. Pulverized solid samples of cortical rabbit bone and a synthetic fluoridated apatite were examined in vitro at variable degrees of hydration. A 9.4 T superconducting spectrometer was used to obtain 31P magic angle spinning NMR spectra and T1 relaxation times. Quantitation was attempted in mixed samples using T1 recovery data. Bone mineral and synthetic apatite could be distinguished by chemical shift and T1 relaxation time in variable hydration states, and were readily differentiated in mixtures by their T1 relaxation time. NMR estimates of relative proportions of components in mixed samples were accurate within 2% of evaluations based on weight. Solid-state 31P NMR therefore provides a suitable method for monitoring the bioabsorption of synthetic apatites.

Animals↗

Quantitative analysis of the solubilizing action of MTAD, sodium hypochlorite, and EDTA on bovine pulp and dentin.

Necrotic bovine pulp and dentin were used in this study as model tissues to represent the organic and inorganic components of the smear layer present in instrumented root canals. The capabilities of endodontic irrigants to dissolve pulverized forms of these tissues were compared. Lyophilized tissue samples were mixed for 2 h at 37 degrees C with MTAD, three concentrations of sodium hypochlorite (NaOCl), 17% EDTA, or isotonic saline. Undissolved tissues were rinsed with water and lyophilized. The change in tissue weight after exposure to an irrigant was measured to quantify solubilization. The results showed that various concentrations of NaOCl removed organic components of pulp and dentin effectively. As pulp solubilizers 5.25% and 2.60% NaOCl were equal (>90%), and 5.25% NaOCl was capable of dissolving virtually the entire organic component of dentin. EDTA was capable of solubilizing inorganic material in dentin and organic material in pulp and likely also in dentin. It dissolved >70% of the dentin and >51% of the pulp. The solubilizing effects of MTAD on pulp and dentin were somewhat similar to those of EDTA. The major difference between the actions of these solutions was a high binding affinity of doxycycline present in MTAD for the dentin.

Animals↗

Clinical and postmortem outcome of "no-reflow' phenomenon in a patient treated with rotational atherectomy.

In the 77-year-old woman described, atherectomy of a circumflex artery with the Rotablator device was complicated by "no reflow." Ten days later, a 75% right coronary artery stenosis was successfully managed by balloon angioplasty. On the following day, acute closure of this vessel resulted in death. Gross examination of the heart showed features of recent posterior infarction, and microscopic study revealed atheroemboli in myocardial arterioles. We conclude that high-speed pulverization of atherosclerotic plaque can cause clinically significant emboli.

Aged↗

Quantitative assessment of free-radical generation during ischemia and reperfusion in the isolated rabbit heart.

BACKGROUND: Several investigators have indirectly implicated oxygen free radicals in reperfusion injury following ischemia in the isolated rabbit heart. METHODS: A quantitative assessment of free-radical production during ischemia and reperfusion was made using electron paramagnetic resonance (EPR) spectroscopy. Serial frozen (77 degrees K) tissue biopsies of the left ventricular wall in isolated rabbit hearts were performed during perfusion, ischemia, and reperfusion. These were pulverized into a fine powder that was filled into EPR tubes. EPR spectra of the tissue were recorded at 77 degrees K. RESULTS: Three predominant signals were seen on EPR analysis: signal A, g = 2.028; signal B, g = 2.005, and signal C, g = 1.940. All three signals were present during the perfusion period and decreased in size during the period of ischemia. After reperfusion, all three signals doubled in size. CONCLUSION: This study lends direct support to the theory that free radicals are generated during myocardial reperfusion and may result in reperfusion injury.

Animals↗

Measles virus prevalence in otosclerotic stapes footplate samples.

HYPOTHESIS: The cause of otosclerosis is still unknown. Persistent measles virus infection of the otic capsule is supposed to be one of the etiologic factors in otosclerosis. Chronic viral antigen expression on the surface of infected cells can induce a secondary autoimmune reaction against the otic capsule. BACKGROUND: In the past 15 years, some reports proposed the possible etiologic role of measles virus in otosclerosis. The presence of measles virus was shown in otosclerotic patients by reverse-transcriptase polymerase chain reaction amplification of the viral RNA, detecting the viral proteins by immunohistochemistry and detecting antimeasles immunoglobulin G in the perilymph samples. Many concerns were elicited by these results. METHODS: Nucleic acid was extracted from pulverized, frozen stapes footplate samples of otosclerotic patients. Measles virus RNA was amplified by reverse-transcriptase polymerase chain reaction: reverse transcription and the first round polymerase chain reaction amplification was performed by heat stable recombinant Thermus thermophilus polymerase, whereas in the nested round, polymerase chain reaction Taq-polymerase was used. Measles virus nucleoprotein RNA-specific oligonucleotide primers were used in these reactions. An Edmonston-type measles virus served as a positive control and cortical bone fragments or stapes superstructures served as negative controls. RESULTS: Among 34 otosclerotic patients, 20 stapes footplate samples contained measles virus RNA. Measles virus RNA was not detected in other bone specimens of the patients. CONCLUSION: The etiologic role of measles virus in the pathogenesis of otosclerosis should be considered. The 14 negative samples may be genetically determined otosclerotic cases.

Adult↗

Antibacterial efficacy of calcium hydroxide and chlorhexidine gluconate irrigants at 37 degrees C and 46 degrees C.

This study investigated the ability of two endodontic irrigants to eliminate Enterococcus faecalis from dentinal tubules, and whether their antimicrobial action was enhanced by heat. The lumens of disks prepared from extracted bovine roots were infected with E. faecalis and incubated for 72 h. Specimens were then filled with saline, 10% calcium hydroxide (Ca(OH)2), or 0.12% chlorhexidine gluconate (CHX) at 24 degrees C or 46 degrees C and incubated at 37 degrees C or 46 degrees C. The samples were then pulverized and plated to quantify residual bacteria. No statistical difference (p > 0.05) in bacterial growth was seen between the two saline groups, or between the two medication groups at a given temperature. CHX and Ca(OH)2 at either temperature produced significantly less growth than either saline group, and CHX or Ca(OH)2 at 46 degrees C produced significantly less growth than either group at 37 degrees C. Heat enhanced the antibacterial action of both experimental irrigants against E. faecalis, but heating saline produced no increase in bactericidal effect.

Animals↗

An in vitro evaluation of the antibacterial efficacy of chlorine dioxide on E. faecalis in bovine incisors.

This study investigated the ability of chlorine dioxide to eliminate Enterococcus faecalis from dentinal tubules of bovine incisors. Thirty-seven extracted bovine incisor roots were sectioned into seventy-four 5 mm disks. Standardized lumens were filled with either sterile Brain Heart Infusion Broth (contamination controls, n = 10) or BHI containing E. faecalis (1.0 x 10 cfu/ml). Disks were incubated in 5% CO2 at 37 degrees C for 72 h. To simulate endodontic instrumentation the lumens were again enlarged. Sixty disks were randomly divided into four experimental groups and filled with one of the following irrigants: 10% Clidox-S (chlorine dioxide), 13.8% BioClenz (chlorine dioxide), 5.25% Clorox, or saline. The disks were incubated for 30 min and were then frozen, pulverized, serially diluted in phosphate buffered saline, and plated on BHI plates in triplicate. Total colony forming units were counted macroscopically. Statistical analysis of the data was performed with a Kruskal-Wallis one-way ANOVA on ranks (p < 0.05, n = 60). Bacterial counts, expressed in log10 cfu/disk were as follows (">" denotes significant differences): Saline > Clidox-S = BioClenz > Clorox. All negative controls were sterile. Chlorine dioxide and NaOCL were both effective in eliminating E. faecalis from the dentinal disks within 30 min.

Animals↗

Pirfenidone inhibits obliterative airway disease in a murine heterotopic tracheal transplant model.

BACKGROUND: Chronic lung allograft rejection in the form of bronchiolitis obliterans syndrome and its histopathologic correlate, obliterative bronchiolitis (OB), are a major source of morbidity and mortality after lung transplantation. Murine heterotopic tracheal transplants into fully allogeneic mismatched recipients develop obliterative airway disease (OAD), which is a suitable model of OB. Using this murine heterotopic tracheal allograft model, we evaluated the effect of pirfenidone, a novel antifibrotic agent, on the development of OAD. METHODS: Mice transplanted with complete MHC-mismatched tracheal allografts received pirfenidone (0.5%) in pulverized food according to different schedules: daily for the first 14 days after transplantation or daily for the duration of the study beginning on posttransplantation days 0, 5, or 10. RESULTS: Mice on a continuous daily regimen of pirfenidone failed to develop evidence of chronic allograft rejection at the termination of the study (60 days). Mice receiving pirfenidone limited to the early posttransplantation period had delayed onset of OAD to 60 days. Forty percent (2/5) of mice receiving a continuous regimen of pirfenidone beginning on day 5 after transplantation had no evidence of OAD at 28 days. However, when the drug was started on day 10, all mice developed OAD by 28 days. CONCLUSIONS: Our results demonstrate a delay of onset or abrogation of OAD when pirfenidone is administered in the early posttransplantation period. These findings suggest that pirfenidone is a candidate drug to be evaluated for prevention of the fibrotic changes seen in OB in human recipients of lung transplants.

Animals↗