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Biomedical subjects

J D Miller

Publications and source records attributed to J D Miller.

At least 91 records · Page 5Linked to original sources

Philopatry of male marine turtles inferred from mitochondrial DNA markers.

Recent studies of mitochondrial DNA (mtDNA) variation among marine turtle populations are consistent with the hypothesis that females return to beaches in their natal region to nest as adults. In contrast, less is known about breeding migrations of male marine turtles and whether they too are philopatric to natal regions. Studies of geographic structuring of restriction fragment and microsatellite polymorphisms at anonymous nuclear loci in green turtle (Chelonia mydas) populations indicate that nuclear gene flow is higher than estimates from mtDNA analyses. Regional populations from the northern and southern Great Barrier Reef were distinct for mtDNA but indistinguishable at nuclear loci, whereas the Gulf of Carpentaria (northern Australia) population was distinct for both types of marker. To assess whether this result was due to reduced philopatry of males across the Great Barrier Reef, we determined the mtDNA haplotypes of breeding males at courtship areas for comparison with breeding females from the same three locations. We used a PCR-restriction fragment length polymorphism approach to determine control region haplotypes and designed mismatch primers for the identification of specific haplotypes. The mtDNA haplotype frequencies were not significantly different between males and females at any of the three areas and estimates of Fst among the regions were similar for males and females (Fst = 0.78 and 0.73, respectively). We conclude that breeding males, like females, are philopatric to courtship areas within their natal region. Nuclear gene flow between populations is most likely occurring through matings during migrations of both males and females through nonnatal courtship areas.

Animals↗

Effect of surgical experience on results of esophagectomy for esophageal carcinoma.

BACKGROUND: Esophagectomy for esophageal cancer is associated with substantial operative morbidity and mortality. The effect of surgical experience on results of esophagectomy has received little attention in the medical literature. METHODS: A retrospective review of esophagectomies for cancer was done. RESULTS: Seventy-four patients underwent esophagectomy by 20 different surgeons. Three surgeons performed 6 or more esophagectomies per year ("frequent" surgeons), whereas the other 17 surgeons performed 5 or fewer esophagectomies per year ("occasional" surgeons). Forty-two patients were operated on by frequent surgeons. There were 3 (7%) anastomotic leaks and no deaths. In 32 patients operated on by occasional surgeons, there were 7 (22%) anastomotic leaks and 7 (22%) operative deaths. The anastomotic leak rates were not significantly different (P < .07), but frequent surgeons had a significantly lower operative mortality (P < .0014). CONCLUSIONS: Esophagectomy for esophageal cancer should be performed by experienced esophageal surgeons with sufficient yearly volume of procedures to maintain competence.

Esophageal Neoplasms↗

Localization of perlecan (or a perlecan-related macromolecule) to isolated microglia in vitro and to microglia/macrophages following infusion of beta-amyloid protein into rodent hippocampus.

The origin of the heparan sulfate proteoglycan (PG), perlecan, in beta-amyloid protein (A beta)-containing amyloid deposits in Alzheimer's disease (AD) brain is not known. In the present investigation we used indirect immunofluorescence, SDS-PAGE, and Western blotting with a specific perlecan core protein antibody to identify possible cell candidates of perlecan production in both primary cell cultures and in a rat infusion model. Double and triple-labeled indirect immunofluorescence was performed on dissociated primary rat septal cultures using antibodies for specific identification of cell types and for perlecan core protein. In mixed cultures of both embryonic day 18 (containing neurons and glia) and postnatal day 2-3 (devoid of neurons), microglia identified by labeling with OX-42 or anti-ED1 were the only cell type also double labeled with an affinity-purified polyclonal antibody against perlecan core protein. Similar immunolabeling of microglia with the anti-perlecan antibody was also observed in purified cultures of post-natal rat microglia. Analyses of PGs from cultured postnatal rat microglia by Western blotting using a polyclonal antibody against perlecan core protein revealed an approximately 400 kDa band in cell layer, which was intensified following heparitinase/heparinase digestion, suggestive of perlecan core protein. Other lower Mr bands were also found implicating either degradation of the 400 kDa core protein or the presence of separate and distinct gene products immunologically related to perlecan. Reverse transcription followed by polymerase chain reaction using human perlecan domain I specific primers demonstrated perlecan mRNA in cultured human microglia derived from postmortem normal aged and AD brain. Following a 1-week continuous infusion of A beta (1-40) into rodent hippocampus, immunoperoxidase immunocytochemistry and double-labeled immunofluorescent studies revealed perlecan accumulation primarily localized to microglia/macrophages within the A beta infusion site. These studies have identified microglia/macrophages as one potential source of perlecan (or a perlecan-related macromolecule) which may be important for the ongoing accumulation of both perlecan and A beta in the amyloid deposits of AD.

Amyloid beta-Peptides↗

Toxic effects of deoxynivalenol on ribosomes and tissues of the spring wheat cultivars Frontana and Casavant.

The toxic effects of the Fusarium graminearum trichothecene toxin deoxynivalenol were determined on ribosomes and leaf tissues of the fusarium head blight-resistant spring wheat cultivar Frontana and the susceptible spring wheat cultivar Casavant. The use of a poly-U-directed 14C-phenylalanine and deoxynivalenol ribosome-binding assays provided evidence of resistance to the protein-synthesis inhibition effects of deoxynivalenol in the head blight-resistant cultivar Frontana. This is probably due to the existence of a mutation in the peptidyl transferase. This cultivar also exhibited resistance to the membrane-damaging properties of this toxin compared to the other cultivar. This report summarizes the evidence for various kinds of "trichothecene tolerance" mechanisms in fusarium head blight-resistant wheat genotypes.

Electrolytes↗

Absence of a synergistic effect between fumonisin B1 and N-nitrosomethylbenzylamine in the induction of oesophageal papillomas in the rat.

Fumonisins and N-nitrosamines (NNO) are suggested risk factors in the development of human oesophageal cancer; exposure to both occurs in high risk populations in Africa and People's Republic of China. The hypothesis that the two would interact in oesophageal carcinogenesis was therefore tested by treating male rats with the known oesophageal carcinogen N-methylbenzylnitrosamine (NMBA), and fumonisin B1 (FB1). The treatment groups were: Group 1, NMBA (2.5 mg/kg) intraperitoneally twice per week from week 2 to 4 inclusive; Group 2, as for group 1 but in addition FB1 (5 mg/kg) daily from weeks 1 to 5 inclusive by gavage; Group 3, FB1 (5 mg/kg) alone daily from weeks 1 to 5 inclusive by gavage, and Group 4, vehicle treatment from week 1 to 5 inclusive. Two of 12 animals in group 1 developed oesophageal papillomas and a further two had oesophageal dysplasia. Data were similar in group 2, animals receiving both NMBA and FB1, with one of 12 animals having papillomas and three of 12 with dysplasia. Sphingolipid biosynthesis was affected in the kidney and slightly in the liver after fumonisin treatment but not in the oesophagus or lung as determined by sphinganine:sphingosine ratios in urine and tissues. These data show that there is no synergistic interaction between NMBA and FB1 in the rat oesophagus when the two compounds are administered together. It is nevertheless important to examine other experimental models and treatment protocols which may be more relevant to the human situation and also to pursue epidemiological investigations of the role of fumonisins in oesophageal cancer.

Animals↗

The use of ergosterol to measure exposure to fungal propagules in indoor air.

This report concerns the development of a method for the measurement of ergosterol in indoor air as a determinant of fungal exposure. Ergosterol was determined in spores of 11 species of Aspergillus, Penicillium, and Cladosporium selected from the most common molds in 400 homes in Ontario. Spore ergosterol content was about 1 microgram/mg, which is the range reported for mycelia, and varied by about 25% for the species tested. Ergosterol was determined in bedroom air samples taken in the winter in homes in southern Ontario. The median ergosterol value corresponded to a total concentration of fungal spores on the order of 10 to 10(2) per m3, in the range for other studies where total and viable propagules were determined by other methods. The sampling of air for ergosterol is a robust method for assessing fungal biomass in air, but provides no information on the species present.

Air Microbiology↗

Effects of alfentanil on cerebral haemodynamics in an experimental model of traumatic brain injury.

Alfentanil is reported to increase intracranial pressure (ICP) after neurotrauma. A direct cerebral vasodilator effect has been postulated. We studied 17 Sprague-Dawley rats allocated to one of three groups. Animals were anaesthetized and their lungs ventilated, and arterial pressure, ICP and/or regional cerebral blood flow (CBF) measurements were undertaken. Group 1 (n = 6) received a severe closed head injury while group 2 (n = 5) received no injury. ICP and mean arterial pressure (MAP) were measured before, during and after rapid infusion of alfentanil 250, 500 and 750 micrograms kg-1. CBF was measured by hydrogen clearance before rapid infusions and at 30-min intervals after starting a subsequent slow infusion of alfentanil 500 micrograms kg-1 h-1. Group 3 (n = 6) underwent CBF measurement only, for comparison with those of groups 1 and 2. They received an injury but no alfentanil. ICP or MAP values did not differ significantly between groups 1 and 2. Rapid i.v. doses of alfentanil produced increases in ICP and reductions in MAP. ICP changes were consistent with a drug effect (P < 0.001) but were small. Reductions in MAP were significant (P < 0.05) and preceded changes in ICP. CBF values were similar and unaffected by slow alfentanil infusion in groups 1 and 2, and did not differ significantly between groups 1 and 3. We conclude that alfentanil did not appear to exert a direct effect on the cerebral circulation. Changes in ICP after rapid infusion were secondary to reductions in SAP. Slow infusion did not cause such changes.

Alfentanil↗

Effects of fumonisin B1 on the immune system of sprague-dawley rats following a 14-day oral (gavage) exposure.

The effects of fumonisin B1 (FB1) on the immune system of Sprague-Dawley rats were investigated. Groups of male and female rats (10 rats/group) were gavaged daily for 14 days with doses of 0, 5, 15, and 25 mg/kg body wt/day and the primary (IgM) response to sheep red blood cells expressed as plaque-forming cell numbers/10(6) spleen mononuclear leukocytes (PFC/10(6) splenocytes) and PFC/spleen was determined. There was a significant dose-related linear trend toward decreased PFC/10(6) splenocytes (p = 0.003) and PFC/spleen cells (p = 0.001) in the male rats. Body weights, expressed as a percentage of the control, were significantly reduced (p = 0.002) in the male rats administered 15 and 25 mg/kg doses. The PFC numbers in female rats were not affected significantly by treatment (p > 0.05). For the remaining immunotoxicity studies, groups of male rats (10 rats/group) were gavaged with FB1 doses of 0, 1, 5, and 15 mg/kg body wt/day for 14 days. There was a weakly significant dose-related trend toward increased numbers of serum immunoglobulin class G (p = 0.04). Also a significant dose-related increase (p = 0.013) in Listeria monocytogenes numbers was observed in the spleen at 24 hr postinfection. Treatment did not have a significant effect on organ weights, hematology, mitogen-induced lymphocyte transformation, calcium mobilization, the numbers of leukocytes and T-lymphocyte subsets, the natural killer cell activity, and phagocytosis (p >/= 0. 05). These observations suggested that FB1 may have indirect consequences for human health and warrant further investigations.

Administration, Oral↗

Comparative study of continuous extrapleural intercostal nerve block and lumbar epidural morphine in post-thoracotomy pain.

OBJECTIVES: To compare the efficacy of continuous extrapleural intercostal nerve block with bupivacaine 0.5% in 1:200,000 epinephrine and continuous lumbar epidural block with morphine in controlling post-thoracotomy pain and to measure serum bupivacaine concentrations during extrapleural infusion. DESIGN: A prospective, randomized, controlled trial. SETTING: St. Joseph's Hospital, Hamilton, Ont., a tertiary care teaching centre. PATIENTS: Sixty-one patients booked for elective thoracotomy were randomized by scaled envelope to two groups. INTERVENTIONS: Group A received a continuous extrapleural intercostal nerve block with bupivacaine 0.5% in 1:200,000 epinephrine as a bolus of 0.3 mL/kg followed by an infusion of 0.1 mL/kg every hour for 72 hours. Group B received a continuous lumbar epidural block with morphine as a bolus of 70 g/kg followed by an infusion of 7 g/kg every hour for 72 hours. MAIN OUTCOME MEASURES: Pain was assessed by a linear visual analogue scale (VAS) pain score. The cumulative amount of "rescue" intravenous morphine used, and serum bupivacaine concentrations were measured as secondary outcomes. RESULTS: Pain control was the same in both groups as assessed by linear VAS score (p = 0.33). The cumulative dose of intravenous morphine for supplemental analgesia was statistically significant between the groups: group A patients used more morphine than group B (p < 0.05). Accumulation of serum bupivacaine was present with no clinical toxicity. CONCLUSIONS: There is no significant difference in the degree of post-thoracotomy pain control measured by the VAS score when analgesia is provided by continuous extrapleural intercostal nerve block with bupivacaine 0.5% in 1:200,000 epinephrine or lumbar epidural block with morphine. Larger amounts of rescue analgesia were used by patients in the continuous extrapleural group with bupivacaine than those in the continuous lumbar epidural block with morphine. Serum bupivacaine concentrations rise without clinical toxicity.

Analgesia, Epidural↗

The CBFbeta subunit is essential for CBFalpha2 (AML1) function in vivo.

The CBFbeta subunit is the non-DNA-binding subunit of the heterodimeric core-binding factor (CBF). CBFbeta associates with DNA-binding CBFalpha subunits and increases their affinity for DNA. Genes encoding the CBFbeta subunit (CBFB) and one of the CBFalpha subunits (CBFA2, otherwise known as AML1) are the most frequent targets of chromosomal translocations in acute leukemias in humans. We and others previously demonstrated that homozygous disruption of the mouse Cbfa2 (AML1) gene results in embryonic lethality at midgestation due to hemorrhaging in the central nervous system and blocks fetal liver hematopoiesis. Here we demonstrate that homozygous mutation of the Cbfb gene results in the same phenotype. Our results demonstrate that the CBFbeta subunit is required for CBFalpha2 function in vivo.

Alleles↗

Prostate-specific antigen expression is regulated by an upstream enhancer.

Prostate cancer can be detected using assays for blood-borne prostate-specific antigen (PSA), which is the clinically most useful diagnostic marker of malignant disease. This paper characterizes the 5 -flanking prostate-specific enhancer which controls expression of the human PSA gene This enhancer, located between -5824 and -3738, is androgen-responsive and requires a promoter for activity. Inductions of 12-100-fold activity occur at 1 nM concentrations of the testosterone analog R1881. The enhancer demonstrated tissue specificity as judged by transfections of several human cell lines. Electrophoretic mobility shift assays comparing nuclear extracts from breast cancer cells MCF-7, and prostate cancer cells LNCaP, showed three regions of prostate-specific binding. These three regions are -4168 to -4797 (region I), -4710 to 4479 (region II), and -4168 to -3801 (region III). Region III contained a putative androgen response element at -4136 that markedly affected activity if mutated. These data suggest that prostate-specific gene expression may involve interaction of prostate-specific proteins or protein complexes with the enhancer in addition to binding of the androgen receptor to androgen response elements.

Androgens↗

Response of growing swine to dietary exposure to pure fumonisin B1 during an eight-week period: growth and clinical parameters.

Consumption of corn or corn-based products contaminated with Fusarium moniliforme/fumonisins has been associated with a variety of animal and human diseases and is a major food/feed safety issue. This study focused on the clinical toxicity and performance parameters in growing swing exposed to low to moderate levels of pure fumonisin B1 (FB.) for 8 weeks. Male (castrated) and female pigs were fed diets containing 0,0.1,1.0, and 10 mg FB1/kg diet (ppm). Weight gains and feed consumption were measured weekly. Blood samples were collected throughout the study, and various clinical and hematological parameters were measured. Because fumonisins are potent inhibitors of sphingolipid biosynthesis, sphinganine and sphingosine concentrations were determined in the liver, lung, and kidney. Organ weights and carcass quality were measured at the end of the trial. In general, male pigs were more adversely affected by FB1 in the diet than females. The average daily gain for males decreased by 8% for pigs fed 1.0 ppm and by 11% at 10.0 ppm, when compared to the control (0 ppm). Males fed 0.1 ppm showed an erratic growth pattern during the first 5 weeks of the experiment. Feed consumption for the same animals was somewhat higher than that of the controls during each of the first 4 weeks but thereafter was 6-7% lower each week as compared to controls. Female pigs fed FB1-diets showed a general enhancement of feed consumption until week 4. Among clinical chemistry parameters, cholesterol increased in males for the 1.0 and 10.0 ppm diets as compared to controls after 2 weeks, while the levels in both sexes were elevated for the 1.0 ppm diet only by the end of the experiment. Serum liver enzyme concentrations were altered during week 2 only. Changes were observed in the weight of the pancreas and adrenals for male pigs fed FB1 diets as compared to controls. The free sphinganine to free sphingosine ratio (biomarker of exposure in FB1-consuming animals) increased in all three organs for the 10 ppm diet, regardless of sex. The study indicated that FB1 can cause different effects at each dose level, at concentrations as low as 0.1 ppm (showing erratic growth) followed by a reduced growth and biochemical abnormalities in blood (1.0 ppm) and sphingolipid alterations in tissues (10.0 ppm). Some of these effects occurred below the exposure level that caused alteration in sphingolipid metabolism.

Adrenal Glands↗

Fumonisin B1 toxicity in male Sprague-Dawley rats.

Male rats were gavaged with fumonisin B1 (FB1) once daily for 11 consecutive days at doses of 0, 1, 5, 15, 35, and 75 mg FB1/kg body weight. Urine osmolality (at 5-75 mg FB1/kg) and organic ion transport in kidney slices (at 5-75 mg FB1/kg) were reduced. Urinary excretion of protein (at 15-75 mg FB1/kg) and of the enzymes LDH (at 5-75 mg FB1/kg), NAG (at 5-75 mg FB1/kg) and GGT (at 15-75 mg FB1/kg) were increased. These findings were indicative of glomerular and tubular toxicity. Histopathologic changes in the kidney consisted of necrosis of tubular epithelia of variable extent accentuated in the inner cortex. These changes were present at 1 and 5 mg FB1/kg and were more pronounced at 15-75 mg FB1/kg. Serum enzymes indicative of hepatotoxicity (ALT, GGT) were elevated compared to controls at 75 mg FB1/kg only. There were noticeable increases in mitotic figures in hepatocytes at 35-75 mg FB1/kg, while single cell necroses were increasingly numerous from 15-75 mg FB1/kg. The kidneys were considered to be the primary target organs in this study.

Acetylglucosaminidase↗

Biological fate of fumonisin B1 in food-producing animals.

The presence of mycotoxins in grains and feedstuffs causes not only animal health problems, but also a valid concern about the transmission of potentially toxic residues into animal-derived products intended for human consumption. In a series of studies at Agriculture and Agri-Food Canada, we investigated the biological fate of fumonisin B1 (FB1) in several food-producing animals (grower pigs, laying hens, dairy cattle), as well as monitored various parameters for evidence of toxicity in these species. In several experiments involving either single-dose protocols (iv, po) or longer-term feeding trials, the pharmacokinetic profiles of FB1 (purity > 95%) in these species were determined, including tissue accumulation and transmission of residues. Toxicological (and economical) implications such as performance (feed consumption, growth), productivity, and carcass quality were also measured when appropriate.

Absorption↗

NMR structural studies of fumonisin B1 and related compounds from Fusarium moniliforme.

Fumonisin B1 (FB1) is the primary mycotoxin produced by Fusarium moniliforme and appears to be responsible for the varied toxigenic effects associated with ingestion of this mold, particularly that of the inhibition of sphingolipid biosynthesis. Understanding the structure and biosynthesis of fumonisins is a key factor in determining structure/activity relationships. To this end, Nuclear Magnetic Resonance (NMR) methods have been used to identify various derivatives of FB1, both naturally occurring and synthetic. With accurate chemical shift assignments, NMR may be used to determine the level of impurities in toxicological grade FB1 preparations. Specifically enriched FB1 was prepared from F. moniliforme cultures using 13C-enriched acetate as well as several 13C-enriched amino acids. 13C NMR analysis indicates that the biosynthesis of fumonisins involves the addition of methionine-derived methyl functions, glutamate-derived tricarballylic ester functions and alanine to an 18 carbon hydrocarbon backbone that is likely polyketide in origin. With the goal of obtaining a crystalline compound for the determination of absolute configuration, several derivatives of FB1 have been prepared, and NMR analysis used to determine the relative and absolute configuration of the 10 stereocenters present in this molecule.

Carcinogens, Environmental↗