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Biomedical subjects

D M Miller

Publications and source records attributed to D M Miller.

At least 73 records · Page 4Linked to original sources
PubMed1998

The Caenorhabditis elegans gene T23G5.5 encodes an antidepressant- and cocaine-sensitive dopamine transporter.

A small subset of neurons in the nematode Caenorhabditis elegans utilizes the catecholamine dopamine (DA) as a neurotransmitter to control or modulate movement and egg-laying. Disruption of DA-mediated behaviors represents a potentially powerful strategy to identify genes that are likely to participate in dopaminergic systems in man. In vertebrates, extracellular DA is inactivated by presynaptic DA transport proteins (DATs) that are also major targets of addictive agents, including amphetamines and cocaine. We used oligonucleotides derived from the C. elegans genomic locus T23G5.5 to isolate and characterize T23G5.5 cDNAs. Our studies predict that mRNAs from this locus encode a 615-amino-acid polypeptide with twelve stretches of hydrophobicity suitable for transmembrane domains, similar to that found in vertebrate catecholamine transporters. The inferred translation product bears highest identity (43-47%) to catecholamine (DA, norepinephrine, epinephrine) transporters within the GAT1/NET gene family and possesses conserved residues implicated in amine substrate recognition. Consistent with these findings, HeLa cells transfected with the C. elegans cDNA exhibit saturable and high affinity DA transport (Km = 1.2 microM) that is dependent on extracellular Na+ and Cl- and blocked by inhibitors of mammalian catecholamine transporters, including norepinephrine transporter- and DAT-selective antagonists, tricyclic antidepressants, and the nonselective amine transporter antagonists cocaine and D-amphetamine. These studies validate the T23G5.5 locus as encoding a functional catecholamine transporter, providing important comparative sequence information for catecholamine transporter structure/function studies and a path to identify regulators of dopaminergic signaling via genetic or pharmacologic manipulation of C. elegans cDNA in vivo.

Animals↗
PubMed1997

Divalent transition metal cations counteract potassium-induced quadruplex assembly of oligo(dG) sequences.

Nucleic acids containing tracts of contiguous guanines tend to self-associate into four-stranded (quadruplex) structures, based on reciprocal non-Watson-Crick (G*G*G*G) hydrogen bonds. The quadruplex structure is induced/stabilized by monovalent cations, particularly potassium. Using circular dichroism, we have determined that the induction/stabilization of quadruplex structure by K+is specifically counteracted by low concentrations of Mn2+(4-10 mM), Co2+(0.3-2 mM) or Ni2+(0.3-0.8 mM). G-Tract-containing single strands are also capable of sequence-specific non-Watson-Crick interaction with d(G. C)-tract-containing (target) sequences within double-stranded DNA. The assembly of these G*G.C-based triple helical structures is supported by magnesium, but is potently inhibited by potassium due to sequestration of the G-tract single strand into quadruplex structure. We have used DNase I protection assays to demonstrate that competition between quadruplex self-association and triplex assembly is altered in the presence of Mn2+, Co2+or Ni2+. By specifically counteracting the induction/stabilization of quadruplex structure by potassium, these divalent transition metal cations allow triplex formation in the presence of K+and shift the position of equilibrium so that a very high proportion of triplex target sites are bound. Thus, variation of the cation environment can differentially promote the assembly of multistranded nucleic acid structural alternatives.

Cations, Divalent↗
PubMed1997

Successful term pregnancy following conservative debulking surgery for a stage IIIA serous low-malignant-potential tumor of the ovary: a case report.

Low-malignant-potential tumors of the ovary can occur in young women of childbearing age. Often these tumors are early stage and confined to the ovary at diagnosis allowing for fertility-preserving surgery. This case report describes a 25-year-old woman who presented with an advanced-stage metastatic LMP tumor and who underwent successful tumor debulking while preserving normal ovarian function. A successful spontaneous pregnancy occurred subsequently and the patient has remained well with 2 years of follow-up.

Adult↗
PubMed1997

Basal cell carcinoma of the vulva: clinical features and treatment results in 28 patients.

OBJECTIVE: To review our experience and that in the recent literature regarding basal cell carcinoma of the vulva to see whether current management guidelines are appropriate. METHODS: Twenty-eight women with basal cell carcinoma of the vulva were seen over 25 years at the BC Cancer Agency. The clinical-pathologic features were tabulated and the outcome was analyzed. RESULTS: The mean age was 74 years, and almost two-thirds were over the age of 70 at diagnosis. Patients typically presented with an irritation or soreness, with a symptom duration ranging from a few months to several years. Most lesions were confined to the anterior half of the vulva, and 23 of the 28 patients had T1 lesions. Wide local excision was the treatment method used most commonly. Only one patient was known to have died from disease metastasis. Ten women had other basal cell carcinomas, either before or after the diagnosis of their vulvar lesions, and in ten patients 11 other malignancies were diagnosed. CONCLUSION: Basal cell carcinoma of the vulva is an extremely uncommon tumor that rarely metastasizes or spreads. Primary treatment should consist of wide local excision and continued follow-up.

Aged↗
PubMed1997

Improving mental well-being in the workplace.

Occupational physicians, who are in a key position to influence company policy on mental health, have limited access to education on mental health. This paper discusses the results of a postal survey of occupational physicians that was designed to identify the mental health information needs of occupational physicians, determine the way in which this information should be disseminated and measure the extent to which occupational physicians encounter mental health problems in the workplace.

Attitude of Health Personnel↗
PubMed1997

A simple method for the continuous noninvasive estimate of cardiac output using the Maxima breathing system. A pilot study.

The Maxima is a new breathing system which makes it possible to monitor cardiac output (Q) noninvasively and to follow trends continuously. When used as a rebreathing system in controlled ventilation, the Maxima selectively eliminates alveolar gas and no fresh gas (VF) is eliminated. Without the need for a mixing chamber, eliminated CO2 (FeCO2) may be measured directly. With steady state conditions, and assuming a respiratory quotient of 1, carbon dioxide production (VCO2) = VF x FeCO2. The indirect Fick principle applied to CO2 exchange (Q = VCO2/ venous-to-arterial CO2 content difference (CaCO2-CvCO2) may be modified to Q = VF x FeCO2/(CaCO2-CvCO2). If VF in the breathing system is adjusted so that FeCO2 is equal to CaCO2-CvCO2, then Q = VF. It was found that, in the presence of a normal haemoglobin (13.3 g.dl-1), Q = VF when VF was adjusted to achieve an FeCO2 value of approximately 4.1%. Cardiac output estimates were compared with those obtained using a thermodilution technique in five patients undergoing coronary artery bypass grafting and in four patients with septicaemia in the ICU. Cardiac outputs were estimated from 1. VCO2 measurements and then 2. VCO2 and haemoglobin (Hb). The result of 28 measurements on nine patients with methods 1. and 2. respectively yielded a correlation with thermodilution measurements: coefficient r = 0.91 and 0.94 with a bias of -10.5% and -0.05% and an accuracy of 14% and 9%.

Capnography↗
PubMed1997

The Groucho-like transcription factor UNC-37 functions with the neural specificity gene unc-4 to govern motor neuron identity in C. elegans.

Groucho and Tup1 are members of a conserved family of WD repeat proteins that interact with specific transcription factors to repress target genes. Here we show that mutations in WD domains of the Groucho-like protein, UNC-37, affect a motor neuron trait that also depends on UNC-4, a homeodomain protein that controls neuronal specificity in Caenorhabditis elegans. In unc-4 mutants, VA motor neurons assume the pattern of synaptic input normally reserved for their lineal sister cells, the VB motor neurons; the loss of normal input to the VAs produces a distinctive backward movement defect. Substitution of a conserved residue (H to Y) in the fifth WD repeat in unc-37(e262) phenocopies the Unc-4 movement defect. Conversely, an amino acid change (E to K) in the sixth WD repeat of UNC-37 is a strong suppressor of unc-37(e262) and of specific unc-4 missense mutations. We have previously shown that UNC-4 expression in the VA motor neurons specifies the wild-type pattern of presynaptic input. Here we demonstrate that UNC-37 is also expressed in the VAs and that unc-37 activity in these neurons is sufficient to restore normal movement to unc-37(e262) animals. We propose that UNC-37 and UNC-4 function together to prevent expression of genes that define the VB pattern of synaptic inputs and thereby generate connections specific to the VA motor neurons. In addition, we show that the WD repeat domains of UNC-37 and of the human homolog, TLE1, are functionally interchangeable in VA motor neurons which suggests that this highly conserved protein domain may also specify motor neuron identity and synaptic choice in more complex nervous systems.

Alleles↗
PubMed1997

Evaluation of low-level aflatoxin in the diet of white-tailed deer.

We evaluated the response of white-tailed deer (WTD) (Odocoileus virginianus) to dietary aflatoxin. Fourteen 4-to-5-mo-old WTD were used in this 8-wk study, conducted between November 1993 and January 1994. Seven animals received a ration containing 800 parts per billion (ppb) total aflatoxin (AF). Seven control animals received the same ration without AF. At 0, 1, 3, 6 and 8 wk, feed consumption, feed conversion, liver enzymes, bile acid levels, and immune function via lymphocyte proliferation assays and delayed type hypersensitivity reactions were determined. At the conclusion of the 8-wk feeding trial, deer were euthanized and necropsied. Clinical illness was not evident in any of the animals, but by the end of the study, AF-fed deer had reduced feed consumption and body weight as compared to control deer; the differences were not statistically significant. The AF-exposed group had a significant increase (P = 0.03) in serum bile acid concentration as compared to control deer. Two AF-exposed deer had gross and histologic hepatic lesions indicative of a mild degenerative hepatopathy. Residues of an aflatoxin metabolite, aflatoxin M1, were found in the livers of all treated animals. No differences in immune function were detected between the two groups. We conclude that consumption of 800 ppb AF in the diet of young WTD over an 8-wk period can produce subclinical hepatic injury.

Administration, Oral↗
PubMed1997

Gliomatosis peritonei with malignant transformation.

A 13-year-old girl underwent a right salpingo-o-ophorectomy and partial omentectomy for an ovarian tumor that on microscopic examination was a grade 1 immature teratoma. Mature glial implants were found in the omentum (Stage III). No additional treatment was given. Ten months later, grade 0 and grade 1 peritoneal glial implants were found on laparotomy. Chemotherapy with four cycles of vincristine/cisplatin/etoposide/bleomycin was administered. A second laparotomy 4 months later showed persistence of grade 0 and grade 1 peritoneal glial implants. The patient was well for the next 7 years, after which time she presented with a pelvic mass that on microscopic examination was a malignant neuroectodermal tumor resembling a glioblastoma multiforme. The tumor did not respond to debulking and chemotherapy, and the patient died 6 months later, 8 years after her initial presentation. This case represents the second report of malignant transformation of peritoneal glial implants.

Adolescent↗
PubMed1996

Influence of GC and AT specific DNA minor groove binding drugs on intermolecular triplex formation in the human c-Ki-ras promoter.

We have used DNase I footprinting and gel shift assays to characterize the interaction of DNA binding drugs mithramycin, distamycin, and berenil with an intermolecular triplex formed by the human c-Ki-ras promoter. A purine-rich triplex-forming oligonucleotide (ODN) forms a stable intermolecular triple helix (triplex) with a homopurine (PR):homopyrimidine (PY) motif in the human c-Ki-ras promoter which contains a 22bp PR:PY region (-328 to -307). This triplex structure is comprised of 15 G.G:C triplets interspersed with 7 T.A:T triplets. Mithramycin binding sites in the human c-Ki-ras promoter encompass most of the triplex target site and three G-C-rich sequences downstream of this triplex-forming region. Mithramycin binding within the c-Ki-ras promoter completely abrogates triplex formation. Furthermore, the addition of mithramycin to pre-formed triplex by c-Ki-ras promoter displaces the major groove bound ODN. Five prominent distamycin binding sites are noted within the c-Ki-ras promoter including the triplex-forming site as well as A-T-rich regions upstream and downstream of the triplex site. Berenil does not bind within the triplex target sequence, and only one berenil binding sequence downstream of the triplex motif was present within the c-Ki-ras promoter fragment. Neither distamycin nor berenil prevents triplex formation, and, furthermore, the addition of either distamycin or berenil to the pre-formed triplex structure did not displace the major-groove-bound third strand. This study demonstrates that GC-specific and AT-specific minor groove ligands differentially affect the intermolecular pur.pur:pyr triplex. A possible biological significance of mithramycin interaction with intramolecular triplex is discussed.

Base Sequence↗
PubMed1996

An automated double staining procedure for bone and cartilage.

Differential skeletal staining is an important part of developmental toxicologic studies. Traditionally these studies have required time-consuming differentiation of one or both stains used and careful attention to the maceration step to prevent specimen destruction. We present a fully automated protocol which does not require differentiation of either dye and incorporates a controlled maceration step which is highly reproducible. This has resulted in high quality staining that is reproducible, stable, and can be done in volume with minimal personnel time. The process involves the staining of skinned, eviscerated specimens fixed in 95% ethanol. Using an automated tissue processor, the specimen is stained in alcian blue for 24 hr, macerated in 3% potassium hydroxide for 24 hr and stained with murexide for 24 hr. The specimens are cleared and preserved in glycerol. Within three days specimens have red stained bone and blue stained cartilage. The procedure was developed using 20-day-old Sprague-Dawley rat fetuses to evaluate the feasibility of using the procedure for teratology studies involving the fetal skeleton. Evenly stained specimens can be examined within three days and stored for years without loss of staining.

Animals↗
PubMed1996

Evaluation of chemical amendments to reduce ammonia volatilization from poultry litter.

Ammonia volatilization from poultry litter often causes high levels of atmospheric ammonia in poultry houses, which is detrimental to both farm workers and birds. Ammonia emissions from houses also aggravate environmental problems, such as acid rain, and result in a loss of fertilizer nitrogen. The objectives of this study were to determine the effect of litter amendments on ammonia volatilization and to determine the effect of these amendments on nitrogen and phosphorus content in litter. The results of this research indicate that alum [Al2(SO4)3.18H2O], ferrous sulfate (FeSO4.7H2O), and phosphoric acid (H3PO4) dramatically reduce ammonia volatilization form litter. The amount of ammonia lost from litter treated with sodium bisulfate (NaHSO4) and a proprietory product made of Ca-Fe silicate with a phosphoric acid coating was not different from the control (untreated litter). Aluminum sulfate (alum) and ferrous sulfate also reduced water soluble P concentrations in litter, whereas phosphoric acid greatly increased water-soluble P levels. The most effective compound evaluated with respect to reducing both ammonia loss and P solubility was alum.

Air Pollution↗
PubMed1996

Human papillomavirus infection: treatment options for warts.

Human papillomavirus can produce verrucae, or warts, on cutaneous and mucosal surfaces. Although the diagnosis is usually straightforward, issues regarding the need for treatment and the choice of treatment are complex. The rationale for the treatment of viral warts is based on the prevention of spread to other areas of the body or to other people; the prevention of irritation, bleeding and other unwanted local effects of the wart; cosmetic and psychosocial considerations, and the possible association with neoplasia. A significant proportion of warts resolve spontaneously within two years. Many treatment options appropriate to primary care are available, including local application of salicylic acid, cantharidin, liquid nitrogen and podophyllum. Direct injection of interferon is a choice in selected cases. The decision to treat and the choice of modality must be individualized.

Humans↗
PubMed1996

Efficient delivery of triplex forming oligonucleotides to tumor cells by adenovirus-polylysine complexes.

Oligonucleotides (ODNs) show great promise in their ability to specifically inhibit single gene expression but must cross the cell membrane, escape the endosomal vesicle, and possibly traverse the nuclear membrane to arrive at their intracellular target molecules. In an attempt to improve the delivery of phosphodiester triplex forming ODNs to malignant cells, we have constructed adenovirus-polylysine (AdpL)-ODN complexes designed to take advantage of the receptor mediated endocytosis of adenoviruses to transfer the ODNs to the cell nucleus. Treatment of several different types of tumor cells in culture by AdpL-ODN complex resulted in superior uptake and persistence of the ODNs compared to both free ODN and cationic lipid-ODN complexes. Nuclear uptake peaks at 4 h and intact ODN persists in the nucleus with a half-life of 12 h. ODN concentrations of 20-70 microM are achieved at 24 h in all monolayer cell lines evaluated to date. ODNs are detected in 50-100% of the total cell population by immunohistochemistry with apparent uptake into vesicles and nuclear localization. Luciferase expression of a co-delivered reporter plasmid suggests that these ODNs are free in the nucleus. AdpL-ODN complexes will provide a valuable tool for delivering unmodified ODNs to the nucleus of malignant cells.

Adenoviruses, Human↗
PubMed1996

Intraventricular cryptococcal cysts.

The case of a 55-year-old immunocompetent woman with central nervous system cryptococcosis and multiple intraventricular cysts is presented. The cysts did not enhance on MR and had signal characteristics similar to cerebrospinal fluid on T1- and T2-weighted images; their intensity was lower than cerebrospinal fluid on proton density-weighted images.

Cerebral Ventricles↗
PubMed1995

Inhibition of in vitro transcription by a triplex-forming oligonucleotide targeted to human c-myc P2 promoter.

Triplex-forming oligonucleotides (TFOs) have been shown to bind in a sequence-specific manner to polypurine/polypyrimidine sequences in several human gene promoters, including the c-myc P1 promoter. TFOs have been shown to inhibit transcription in vitro and the expression of target genes in cell culture. The human c-myc protooncogene contains a 23 base pair purine-pyrimidine-rich motif (-62 to -40) within its predominant promoter, P2, that is a potential target for purine-purine-pyrimidine triplex formation. Using electrophoretic mobility shift analysis (EMSA) and competition experiments, we have demonstrated that a MAZ (myc-associated zinc finger protein) consensus sequence is capable of competing with the purine-pyrimidine motif for the binding of a HeLa nuclear protein. We have shown the formation of an intermolecular triplex using a 23-base purine-rich oligonucleotide antiparallel to the purine-rich target sequence. DNase I footprinting was performed to confirm the exact location of triplex formation. Triplex formation by this oligonucleotide prevents binding of a HeLa nuclear protein (presumably MAZ) to the target site. We have also shown that the P2-targeted TFO is a potent and specific inhibitor of c-myc transcription in vitro. These data demonstrate that this novel TFO inhibits transcription of the c-myc P2 promoter. We propose that the P2-targeted TFO has its effect by blocking the binding of the regulatory factor MAZ.

Base Sequence↗