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J W Allen

Publications and source records attributed to J W Allen.

At least 91 records · Page 5Linked to original sources
PubMed1996

In vivo genotoxicity of dichloroacetic acid: evaluation with the mouse peripheral blood micronucleus assay and the single cell gel assay.

Chlorination is a widely used method for disinfection of drinking water supplies. Reaction of chlorine with naturally present organic compounds can result in toxic by-products. One major disinfection by-product from the chlorination of drinking water is dichloroacetic acid (DCA). This chemical has been shown to be carcinogenic in rodents, yet little genotoxicity data are available to assess the possible role of DNA and/or chromosomal damage in this process. We have used the peripheral blood erythrocyte micronucleus (MN) assay and the alkaline single cell gel electrophoresis (SCG) technique to investigate the in vivo genotoxicity of DCA in bone marrow and blood leukocytes, respectively. The MN assay detects chromosome breakage and/or malsegregation, while the SCG assay detects DNA damage (e.g., single strand breaks, alkali-labile sites, crosslinking). Mice were exposed to this compound in drinking water, available ad libitum, for up to 31 weeks. Our results show a small but statistically significant dose-related increase in the frequency of micronucleated polychromatic erythrocytes (PCEs) after subchronic exposure to DCA for 9 days. In addition, at the highest dose of DCA tested (3.5 g/l), a small but significant increase in the frequency of micronucleated normochromatic erythrocytes (NCE) was detected following exposure for > or = 10 weeks. Coadministration of the antioxidant vitamin E did not affect the ability of DCA to induce this damage, indicating that the small induction of MN by DCA was probably not due to oxidative damage. Based on the lack of any difference observed in the proportion of kinetochore-positive micronuclei between the treated and control animals, we interpret MN as arising from clastogenic events. The SCG technique suggested the presence of DNA crosslinking in blood leukocytes in mice exposed to 3.5 g/l DCA for 28 days. These data provide evidence that DCA may be an extremely weak inducer of chromosome damage when provided to mice in drinking water under conditions which lead to increased levels of tumors.

Animals↗
PubMed1996

Micronuclei induced in round spermatids of mice after stem-cell treatment with chloral hydrate: evaluations with centromeric DNA probes and kinetochore antibodies.

The chromosomal effects of chloral hydrate (CH) on germ cells of male mice were investigated using two methods to detect and characterize spermatid micronuclei (SMN); (a) anti-kinetochore immunofluorescence (SMN-CREST) and (b) multicolor fluorescence in situ hybridization with DNA probes for centromeric DNA and repetitive sequences on chromosome X (SMN-FISH). B6C3F1 mice received single intraperitoneal (i.p.) injections of 82.7, 165.4, or 413.5 mg/kg and round spermatids were sampled at three time intervals representing cells treated in late meiosis, early meiosis, or as spermatogonial stem cells. No increases in the frequencies of SMN were detected for cells treated during meiosis using either SMN-CREST or SMN-FISH methods. After spermatogonial stem-cell treatment, however, elevated frequencies of SMN were detected by both methods. With SMN-FISH, dose trends were observed both in the frequencies of spermatids containing micronuclei and in the frequency of spermatids carrying centromeric label. These findings corroborate the recent report by Allen and colleagues [Allen JW et al.(1994): Mutat. Res. 323:81-88] that CH treatment of spermatogenic stem cells induced SMN. Furthermore, our findings suggest that chromosomal malsegregation or loss may occur in spermatids long after CH treatment of stem cells. Further studies are needed to understand the mechanism of action of the CH effect on stem cells and to determine whether similar effects are induced in human males treated with CH.

Anesthetics, Intravenous↗
PubMed1996

Meiotic recombination and germ cell aneuploidy.

Data on human trisomic conceptuses suggest that the extra chromosome commonly has a maternal origin, and the amount and position of crossing-over on nondisjoined chromosomes is commonly altered. These observations may provide important clues to the etiology of human germ cell aneuploidy, especially in regard to evaluating whether environmental factors play a role. There is concordance of effects of environmental agents on fungi, plants, and animals, which suggests that the overall process of meiosis is well conserved and that chemical and physical agents can affect meiotic recombination, leading to aneuploidy. It seems likely that meiosis in humans will fit the general pattern of meiosis in terms of sensitivity to radiation and chemicals. Thus studies on other organisms provide some insight into the procedures necessary for obtaining useful human data. For example, frequencies of spontaneous meiotic recombination are not uniform per physical length in Drosophila, and different regions of a chromosome respond differently to treatment. Treatments that relieve constraints on the distribution of meiotic exchange, without changing greatly the overall frequency of exchange, may increase the number of univalents and give the impression that there are chromosome-specific responses. Recombination studies that monitor one or a few relatively short genetic regions may also give a false impression of the effects of a treatment on recombination. In addition, meiotic mutants in Saccharomyces and Drosophila highlight a number of processes that are important for production of an exchange event and the utility of that event in the proper segregation of both homologues and sisters. They also suggest that tests for pairing at pachytene, chiasmata at diplotene, and genetic crossing-over may give different results.

Aneuploidy↗
PubMed1996

HSP70-2 is part of the synaptonemal complex in mouse and hamster spermatocytes.

Mouse spermatogenic cells are known to express HSP70-2, a member of the HSP70 family of heat-shock proteins. The purpose of the present study was to characterize further the expression and localization of HSP70-2 in meiotic cells of mice and hamsters. After separating mouse spermatogenic cells into cytoplasmic and nuclear fractions, proteins were separated by two-dimensional gel electrophoresis and detected with HSP-specific antibodies. Of several HSP70 proteins identified in the cytoplasm, only HSC70 and HSP70-2 were also detected in the nucleus. Immunocytological analyses of spermatocyte prophase cells revealed that HSP70-2 was associated with the synaptonemal complex. Surface-spread synaptonemal complexes at pachytene and diplotene stages labeled distinctly with the antiserum to HSP70-2. Synaptonemal complexes from fetal mouse oocytes failed to show any evidence of HSP70-2. Reverse-transcriptase-polymerase chain reaction (RT-PCR) analyses of gene expression confirmed this sex specificity; Hsp70-2 mRNA was detected in mouse testes, but not ovaries. These findings are suggestive of a previously unsuspected sexual dimorphism in structure and/or function of the synaptonemal complex.

Animals↗
PubMed1996

Spermatid micronucleus analysis of aging effects in hamsters.

Spermatid micronuclei (MN) from Armenian hamsters in different age groups were compared with regard to frequencies and kinetochore status (presence or absence) as determined with immunofluorescent staining. Six thousand cells analyzed from each of fifteen young animals (3 months) revealed a group mean frequency of 0.45 MN/1000 spermatids; kinetochore staining was uniformly negative. Six thousand cells scored from each of fifteen older animals (2 years) revealed a group mean frequency of 1.00 MN/1000 spermatids. Most of the MN in these animals were negative for kinetochore staining, although a significant representation of MN with positive kinetochore staining was also observed. The results indicate that frequencies of spermatid MN increase with advancing age, and suggest that the increase is due to significant elevations in both chromosome breakage and chromosome loss.

Aging↗
PubMed1996

Hemolytic anemia does not increase entry into, nor alter rate of clearance of bilirubin from rat brain.

The risk of bilirubin encephalopathy is believed to be increased in hemolytic relative to nonhemolytic jaundice. Young SPRD rats were injected with either acetylphenylhydrazine (APHZ, n = 22) 75 mg/kg or an equivalent volume of the solvent (control, n = 22) intraperitoneally for 3 successive days. One the 4th day, hyperbilirubinemia was induced by a 5-min intravenous infusion of 50 mg/kg bilirubin. After sacrifice (at 10 or 60 min) the brain vessels were flushed in situ with ice-cold saline. Serum bilirubin was 600 +/- 108 mumol/l (mean +/- SD) at 10 min, and 295 +/- 98 mumol/l at 60 min. Hematocrit was significantly reduced in the APHZ rats versus controls (26 +/- 3 vs. 41 +/- 3, p < 0.0001). The concentration of bilirubin in brain was determined by acid chloroform extraction and diazotization. The brain bilirubin values were 5.5 +/- 1.5 versus 5.4 +/- 2.1 mmol/g at 10 min (APHZ vs. control), and 1.3 +/- 0.6 versus 0.8 +/- 0.7 nmol/g at 60 min. The half-life of bilirubin in brain was calculated with an exponential fitting program. The half-life of bilirubin in brain for the two groups was 24.3 +/- 21.9 versus 18.5 +/- 28.3 min. There were no significant differences between the groups in either of these measures. We conclude that in young rats, hemolytic anemia does not increase the acute entry of bilirubin into brain, nor does it affect the clearance of bilirubin from brain.

Anemia, Hemolytic↗
PubMed1996

Bilirubin-oxidizing activity in rat brain.

Bilirubin may be cleared from the brain by transport across the blood-brain barrier and by a 'sink effect' into the cerebrospinal fluid. However, there is also evidence to suggest that bilirubin may be metabolized in the brain by a process of oxidation. The purpose of this study was to confirm the existence of bilirubin metabolism in the brain and to examine the possible contribution of such an activity to the bilirubin staining pattern characteristic of kernicterus. Mitochondrial membrane fractions were prepared in 0.32 M sucrose from whole rat brains as well as brain regions. The change in optical density of a bilirubin solution at 440 nm was measured over time following addition of the mitochondrial suspension. Our results confirmed the existence of a bilirubin-metabolizing activity in brain mitochondrial membranes. This activity could be removed by heating the mitochondrial suspension and had a definable temperature and pH maxima. The rate of oxidation of bilirubin ranged from 109 to 164 pmol/min/mg protein. There were significant differences between rat brain regions in the ability to oxidize bilirubin. However, these differences could not explain the kernicterus staining phenomenon, because the highest activities were found in brain regions which are more heavily stained in kernicterus. We conclude that bilirubin is metabolized by brain mitochondrial membranes at a rate that would appear to represent a biologically meaningful contribution to bilirubin clearance from the brain. In the present model, differences in such an activity between brain regions do not appear to be compatible with a kernicteric staining pattern.

Animals↗
PubMed1995

Immediate and late outcome of percutaneous balloon mitral valvotomy in patients with significantly calcified valves.

We assessed immediate and late outcome in 55 patients with significantly calcified valves (group 1) after balloon mitral valvotomy and compared the results with those from 60 patients with noncalcified or minimally calcified valves (group 2). After valvotomy, mitral valve area increased from 1.03 +/- 0.30 cm2 to 1.64 +/- 0.35 cm2 (p = 0.0001) by echo planimetry in group 1 but was significantly smaller than the mitral valve area in group 2 after valvotomy (1.94 +/- 0.38 cm2; p = 0.0001). At a mean follow-up period of 30 months (range 2 to 81 months), 51% of patients in group 1 and 83% in group 2 were symptom free (p = 0.0002). In group 2, 15 (27%) patients and in group 2, 4 (7%) patients had cardiac events (p = 0.003). The risk ratio for cardiac events was 4.3 times greater in group 1 than in group 2. In group 1, the risk ratio for cardiac events was 3.2 times higher in patients age > or = 65 years and in patients with atrial fibrillation. The 6-year cumulative cardiac event-free survival rate was 64% in group 1 and 90% in group 2 (p = 0.005). In 75 (65%) patients who had follow-up echocardiographic study (35 in group 1 and 40 in group 2), mitral valve area decreased to 1.48 +/- 0.42 cm2 at follow-up in group 1 (p < 0.01) and to 1.77 +/- 0.50 cm2 in group 2 (p = 0.3). Restenosis occurred in 16 (46%) of 35 patients in group 1 and 10 (25%) of 40 in group 2 (p = 0.06).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

Comparison of late outcome between Inoue balloon and double-balloon techniques for percutaneous mitral valvotomy in a matched study.

The follow-up results between Inoue balloon (n = 43, group 1) and double-balloon (n = 43, group 2) mitral valvotomies were compared in a patient-to-patient matched study. Matching was based on patients' age, mitral valve echo score, fluoroscopic calcification, mitral valve area before valvotomy, and follow-up period. The mean follow-up period was 13 +/- 9 months for both groups. At follow-up, 72% of patients were symptom free in each group; 2 (4.7%) patients in group 1 and 1 (2.3%) patient in group 2 underwent mitral valve replacement surgery. Of these 3 patients, 2 died after surgery, 1 from each group. The mitral valve area by Doppler was 1.8 +/- 0.3 cm2 in group 1 and 1.8 +/- 0.4 cm2 in group 2 (p = 0.7); the area by echo planimetry was 1.7 +/- 0.3 cm2 in group 1 and 1.8 +/- 0.3 cm2 in group 2 (p = 0.3) at follow-up. Restenosis occurred in 5 (12%) patients from each group. The cumulative restenosis-free rate was 96% at 1 years, 78% at 2 years, and 58% at 3 years in group 1 and 98%, 76%, and 51%, respectively, in group 2 (p = 0.8). Balloon selection did not appear to influence the clinical outcome at follow-up. We conclude that both the inoue balloon and the double-balloon valvotomies are effective therapies with comparable follow-up results for patients with mitral stenosis.

Adult↗
PubMed1995

Germ line specific factors in chemical mutagenesis.

Chemical mutagenesis test results have not revealed evidence of germ line specific mutagens. However, conventional assays have indicated that there are male-female differences in mutagenic response, as well as quantitative/qualitative differences in induced mutations which depend upon the particular cell stage exposed. Many factors inherent in the germ line can be speculated to influence chemical transport to, and interaction with, target cell populations to result in mutagenic outcomes. The level of uncertainty regarding the general operation of such factors, in combination with the limited availability of chemical test data designed to address comparative somatic and germ cell mutagenesis, leaves open the question of whether there are mutagens specifically affecting germ cells. This argues for a conservative approach to interpreting germ cell risk from somatic cell mutation analysis.

Animals↗
PubMed1994

Ethnomycology, biochemistry, and cultivation of Psilocybe samuiensis Guzmán, Bandala and Allen, a new psychoactive fungus from Koh Samui, Thailand.

Several specimens of Psilocybe and Copelandia species in Koh Samui, Thailand were recently collected for herbarium deposit and scientific study. This paper presents an ethnomycological and biochemical study of one of the species; P. samuiensis Guzmán, Bandala and Allen, a new psychoactive gill fungus reported from Thailand. Mycelium for the cultivation of P. samuiensis was obtained on 6% malt agar from the spores of a dried specimen. The growth of P. samuiensis was similar to that of P. tampanensis Guzmán and Pollock, but more rapid than the mycelium of P. semilanceata (Fr.:Sacc.) Kumm. Laboratory analyses indicates that the alkaloid content in cultured fruit bodies of P. samuiensis is of the same order of magnitude as that found in naturally occurring mushrooms of this species. HPLC analyses of both naturally occurring and in vitro cultivated fruit bodies of P. samuiensis revealed high concentrations of psilocybin and psilocin. Small amounts of baeocystin were also detected. Psilocybin levels varied from 0.23% up to 0.90%. The psilocybin content was highest in the caps. Psilocybin was also found in the cultured non-bluing mycelia of P. samuiensis and varied from 0.24% to 0.32% dry weight. The relative alkaloidal content of psilocybin, psilocin, and baeocystin found in P. samuiensis was similar to that measured in many other psychoactive fungi species, but completely different from that found in P. semilanceata.

Agaricales↗
PubMed1994

Low-dose combined therapy with fluvastatin and cholestyramine in hyperlipidemic patients.

OBJECTIVE: To compare the low-density lipoprotein (LDL) cholesterol-lowering efficacy of low-dose combinations of cholestyramine and fluvastatin. DESIGN: Randomized, double-blind, parallel group, placebo-controlled trial with a 24-week double-blind treatment period divided into three phases. SETTING: Office-based clinics. PATIENTS: Hypercholesterolemic, with LDL cholesterol of 4.14 mmol/L or greater (> or = 160 mg/dL) and plasma triglycerides of 3.39 mmol/L or less (< or = 300 mg/dL). Four hundred sixty patients were screened; 224 patients were randomized into a double-blind treatment period; 203 completed the study; 6 dropped out because of adverse events. INTERVENTION: Patients were treated with 10 mg or 20 mg of fluvastatin alone, 8 g or 16 g of cholestyramine alone, or combinations of these fluvastatin and cholestyramine dosages (six treatment groups). MEASUREMENTS: Changes in lipid variables, particularly LDL cholesterol. RESULTS: The 10-mg and 20-mg fluvastatin monotherapy groups showed considerable reductions in LDL cholesterol initially (-20.1% [SD, 8.8%] and -20.2% [SD, 10.1%], respectively); these reductions were maintained. Reductions in LDL cholesterol that resulted from the addition of cholestyramine, 8 g/d, to 10 mg of fluvastatin and 20 mg of fluvastatin were greater than those observed with monotherapy (10-mg fluvastatin--[10-mg fluvastatin plus cholestyramine], 9.1%; 95% CI, 3.8% to 14.4%) and 20-mg fluvastatin--[20-mg fluvastatin plus cholestyramine], 11.6%; CI, 6.5% to 16.8%). The increase in cholestyramine dose to 16 g/d in the three combination groups provided only a modest additional response. CONCLUSIONS: Low-density lipoprotein cholesterol reductions of about 25% to 30% can be achieved with low-dose combination therapy with fluvastatin and cholestyramine. The addition of low-dose resin appears to produce greater overall cholesterol reduction than does a simple doubling of the fluvastatin dosage. The low-dose combination treatment was highly successful in achieving the goals of the National Cholesterol Education Program guidelines.

Anticholesteremic Agents↗
PubMed1994

Centromere analysis of micronuclei induced by 2-aminoanthraquinone in cultured mouse splenocytes using both a gamma-satellite DNA probe and anti-kinetochore antibody.

We have tested 2-aminoanthraquinone (2-AAQ) as a potential aneugen in a cytokinesis-blocked mouse splenocyte micronucleus (MN) assay. Binucleated cells (BNC) were evaluated for MN, and the MN were further probed with two indicators of centromere presence: an anti-kinetochore autoantibody and a DNA probe for the mouse gamma-satellite locus. A dose-dependent increase in the frequency of BNC with MN was observed. At the highest 2-AAQ concentration (10 micrograms/ml), the frequency of BNC containing MN was increased greater than 10-fold over background. Both centromere-positive and centromere-negative MN were significantly increased. At least 62% of MN at all 2-AAQ doses were positive for the gamma-satellite DNA probe, while 30-53% were labeled with the antikinetochore antibody. In contrast with the 2-AAQ results, after treatment with the aneugen demecolcine (positive control), greater than 80% of MN labelled positive with both probes. This discordance in the results with the two probes after 2-AAQ exposure suggests that the mode of action of this chemical may be as an aneugen by disruption of the kinetochore proteins, as a clastogen with a preferential cleavage site at or near the gamma-satellite locus, or both. Our results also suggest that the use of either of these probes individually may not be an adequate measure of centromere presence. Nevertheless, positive results for both markers provides strong evidence that 2-AAQ is aneugenic.

Animals↗
PubMed1994

Thermodilution left-sided cardiac output for valve area determination after balloon mitral valvotomy.

This study was conducted to establish the validity of left-sided cardiac output measurement with a Swan-Ganz catheter and assess its accuracy in estimating mitral valve area (MVA) by the Gorlin formula. The use of right-sided cardiac output after balloon mitral valvotomy (BMV) can give inaccurate measurements for Gorlin-derived MVA because of the atrial septal defect (ASD) created during the procedure. The left-sided cardiac output was measured with a Swan-Ganz catheter (proximal port in the left atrium and then in the left ventricle, and distal port in the ascending aorta) in 10 consecutive patients before and after BMV. Gorlin-derived MVA cardiac output by this method was compared with (1) Gorlin-derived MVA by means of right-sided cardiac output with and without balloon occlusion of the ASD and (2) MVA measured by echocardiography. Before BMV, a close agreement with a good correlation between left-sided and right-sided cardiac output was found (r = 0.83, p = 0.006). Furthermore, Gorlin-derived MVA by cardiac output with either method was comparable with valve area by echo. After BMV, left-sided cardiac output correlated well (r = 0.92, p = 0.0002) and was comparable with right-sided cardiac output with occlusion of the ASD (mean difference 0.17 +/- 0.49 L/min, p = 0.3) but was significantly lower than the value obtained with open ASD (mean difference 0.93 +/- 0.77 L/min, p = 0.004). Comparison of the correspondent MVAs yielded similar results. Gorlin-derived MVA with left-sided cardiac output and MVA by echo were also similar.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiac Output↗
PubMed1994

Synaptonemal complex aberrations in the pseudoautosomal region of X, Y chromosomes in irradiated hamsters.

The effects of X-radiation, bleomycin and amsacrine (m-AMSA) on the meiotic chromosomes of male Armenian hamsters were determined by electron microscopic analysis of synaptonemal complex (SC) damage. Pachytene stage cells were analyzed 5 or 6 days following their treatment at putative preleptotene-leptotene stages of meiosis. Of the multiple types of SC aberrations observed to be significantly increased over control levels, lateral element breakage and synaptic anomalies were most prevalent. The focus of these studies was on the sex chromosomes which, in the Armenian hamster, reveal an unusually well-defined pseudoautosomal region. In the XY pair, radiation and chemical treatments caused certain forms of structural and synaptic anomalies which appeared to be preferentially localized to telomeric and/or crossover regions. The nature of these specific aberrations, involving breakage, bridge formation and asynapsis, is not well understood; however, their distributions are suggestive of possible relationships with sites and processes of crossing over.

Amsacrine↗
PubMed1994

Late clinical and echocardiographic follow up after percutaneous balloon dilatation of the mitral valve.

OBJECTIVE: To assess the outcome after attempted percutaneous balloon dilatation of the mitral valve in patients with severe mitral stenosis between February 1986 and June 1992. DESIGN: Clinical state, mitral valve area, and restenosis at follow up were analysed. Mitral valve area as determined by the Gorlin formula, planimetry, and Doppler methods was compared before and after dilatation and at follow up. SETTING: University hospital. PATIENTS: 176 patients had serial clinical and Doppler echocardiographic follow up and 44 of them also underwent recatheterisation. RESULT: At follow up 93% of patients were in New York Heart Association functional class I or class II. Mitral valve area (planimetry) increased from 0.97(0.24) cm2 before to 1.86(0.39) cm2 after dilatation (p = 0.0001) and then decreased to 1.72(0.39) cm2 at follow up (p < 0.001); mitral valve area (Doppler) increased from 1.01 (0.24) to 1.89 (0.42) cm2 (p = 0.0001) and then decreased to 1.78(0.40) cm2 (p < 0.05). The overall restenosis rate was 15% and over 90% of the patients were free from cardiovascular events. Age, valvar calcification, echocardiographic score, and mitral valve area after dilatation were found to be determinant predictors of restenosis. In patients who underwent recatheterisation, mitral valve area by the Gorlin method at follow up was comparable with that by planimetry and Doppler methods whereas a significant discrepancy between the three methods was noted immediately after dilatation. CONCLUSION: Balloon dilatation of the mitral valve provided sustained anatomical and functional improvement in over 80% of patients at late follow up. Older age, heavy calcification, high echocardiographic score, and suboptimal immediate results are significant predictors of restenosis. Doppler echocardiographic examination is the procedure of choice for follow up evaluation.

Adolescent↗