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Biomedical subjects

C L Smith

Publications and source records attributed to C L Smith.

At least 91 records · Page 5Linked to original sources
PubMed1998

Current perspectives on pain upon injection of drugs.

A limitation in the administration of parenteral products is the pain caused upon injection. Injection site pain has been predominately associated with intravenous, intramuscular, and subcutaneous administration. It becomes important for the formulation scientist to have a basic understanding of the physiology underlying the pain process, as well as the pharmaceutical factors associated with injection site pain. Initially, this review will provide the reader with a primer on the mediation of pain in the periphery and a compilation of those drugs that have been associated with pain on injection. In addition, this review will present important considerations and general formulation approaches or methods that have been used to overcome pain on injection. Finally, a brief overview of the various experimental systems used to investigate injection site pain is discussed.

Humans↗
PubMed1998

Cross-talk between peptide growth factor and estrogen receptor signaling pathways.

The classical receptor for estradiol is a member of a super-family of nuclear receptors that function as hormone-regulated transcription factors. The ability of the estrogen receptor (ER)-alpha to activate target gene transcription is mediated by two transcriptional activation functions (AF): AF-1 located in the amino-terminal domain and AF-2 found in the carboxyl-terminal portion of the molecule. The ligand binding domain overlaps AF-2, and upon estrogen binding this region undergoes a conformational change that enables it to contribute to the receptor's transcriptional activity. ER activation is accompanied by increased phosphorylation, and in the absence of ligand, activators of protein kinase A or inhibitors of protein phosphatases are able to stimulate ER-dependent gene expression. More importantly, polypeptide growth factors, such as epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I), also stimulate the ER's transcriptional activity in an estrogen-independent manner. The AF-1 domain appears to be required for activation by EGF and IGF-I, and point mutation of a single phosphorylation site located within this domain inhibits the ability of growth factor to activate the ER. Thus, steroid receptor function may be regulated by estrogenic ligands as well as by pathway "cross-talk" from membrane receptors for growth factors.

Animals↗
PubMed1998

An examination of radiation exposure to clinical staff from patients implanted with 137Cs and 192Ir for the treatment of gynecologic malignancies.

The treatment of cancers by placement of radioactive materials within or adjacent to a tumor is clinically known as brachytherapy. Although the intracavitary treatment of gynecologic cancers using 137Cs has been in widespread use for nearly a century, interstitial techniques using 192Ir have developed within the last decade. Both procedures are performed as temporary implants and typically require a hospital stay of approximately 48 h. Significant differences in source strengths, loading conditions, and patient care requirements are visible between the intracavitary and interstitial techniques. Facilities that are experienced in the use of intracavitary techniques may lack clinical experience in the use of newer interstitial procedures. An examination of radiation exposure to the radiation oncology staff administering the treatment, exposure rate at frequently occupied points in the patient room, and exposure to the nursing staff will be of value to those institutions considering integration of interstitial brachytherapy techniques into their department.

Body Burden↗
PubMed1998

Renal stone analysis: is there any clinical value?

It has usually been assumed that the analysis of recovered renal stones should be routinely done. This concept has recently been challenged. Very little work has been done to correlate analytical results with clinical findings and the impact of this information on clinical management. This paper reviews current concepts of stone formation, how this information can be used to interpret stone analysis results and how stone analyses results can be used in clinical-decision-making.

Humans↗
PubMed1998

Local therapy for cytomegalovirus retinitis.

OBJECTIVE: To examine the role of intraocular therapy in the management of cytomegalovirus (CMV) retinitis associated with AIDS. DATA SOURCES: A MEDLINE search was conducted for the years 1980-1997. In addition, relevant articles were cross-referenced to screen for additional information. The AIDS/HIV Treatment Directory was searched for information on ongoing studies. STUDY SELECTION/DATA EXTRACTION: Data regarding the use of local antiviral therapy for CMV retinitis are cited. Emphasis was placed on randomized, controlled trials, but descriptive studies are also included. DATA SYNTHESIS: Intraocular drug administration is an alternative therapy for CMV retinitis that avoids some of the disadvantages associated with systemic treatment. Intravitreal ganciclovir 200-2000 micrograms once weekly has been studied in a number of nonrandomized studies. Although initially effective, intravitreal ganciclovir is associated with a significant relapse rate and development of contralateral CMV retinitis. Intraocular ganciclovir implants offer the advantage of less frequent interventions and constant drug concentrations in the vitreous. Time to progression is significantly longer in patients receiving implants versus intravenous therapy; however, there is a higher incidence of contralateral eye retinitis and extraocular CMV disease with the implants. Currently, the intraocular implant is being studied in combination with oral ganciclovir to decrease the incidence of systemic CMV disease. Foscarnet and cidofovir have also been administered intravitreally for CMV retinitis. Cidofovir may offer the advantage of a long intracellular half-life, which would allow infrequent dosing; however, further study is needed to determine a safe and effective intraocular dosage. CONCLUSIONS: Systemic therapy continues as standard management for CMV retinitis. Local therapy has some advantages and disadvantages, but larger, randomized, controlled trials comparing systemic therapy with local therapy must be completed to define its exact role. Data from an ongoing trial of local plus oral therapy will better define this role.

AIDS-Related Opportunistic Infections↗
PubMed1998

In vitro glucocorticoid receptor binding and transcriptional activation by topically active glucocorticoids.

Mometasone furoate (MF, CAS 83919-23-7, Sch 32088), budesonide (BUD, CAS 51372-29-3), fluticasone propionate (FP, CAS 80474-14-2), and triamcinolone acetonide (TA, CAS-76-25-5) are corticosteroids that are either currently available or under development for allergic rhinitis and asthma. The relative affinity of these drugs for the glucocorticoid receptor and their ability to stimulate glucocorticoid receptor-mediated transactivation of gene expression were analyzed. All of the test compounds had a higher affinity for the recombinant glucocorticoid receptor than the reference glucocorticoid receptor ligand, dexamethasone (DEX, CAS 50-02-2). In addition, all compounds showed greater potency than dexamethasone in stimulating transcription of a synthetic target gene regulated by a glucocorticoid response element. Of the compounds tested, mometasone furoate had the highest relative binding affinity for the glucocorticoid receptor, followed by fluticasone propionate, budesonide, and triamcinolone acetonide. Similarly, mometasone furoate was the most potent stimulator of glucocorticoid receptor-mediated transactivation of gene expression, followed by fluticasone propionate, tri-amcinolone acetonide, and budesonide. These in vitro studies provide a sensitive means to compare the potency of glucocorticoids and may reliably predict the in vivo topical potency of these drugs.

Animals↗
PubMed1998

Postcastration eventration in 18 horses: the prognostic indicators for long-term survival (1985-1995).

Castration of horses is considered a common and routine surgical procedure, but the potential for complications is high. By far the most serious of these is eventration. The objectives of this study were to determine the long-term survival of horses undergoing surgical treatment of indirect (1) inguinal eventration of the small intestine following castration, and to identify prognostic indicators for survival. The case records of 18 horses undergoing surgical treatment of postcastration eventration (PCE) between 1985 and 1995 were reviewed. Follow-up information was obtained by telephone interviews 2 to 13 y postoperatively. A Cox proportional hazard regression model was fitted to determine which clinical features were of significant influence to survival. Clinical features with a significant negative influence on survival were an inguinal surgical approach for correction, an increased length of prolapsed bowel, and performance of bowel resection and anastomosis. Significant postoperative complications developed in 89% of cases; 44% of cases in the "inguinal" surgical approach group developed peritonitis, compared with 10% in the "midline" approach group. Of all horses in this study, 72% were discharged from the hospital; however, only 40% of horses in the inguinal approach group were discharged. The long term survival rate (> 1 y) for all horses in this study was 44%, with a median survival time of 3-1/2 mo.

Animals↗
PubMed1997

Principal transcription sigma factors of Pseudomonas putida strains mt-2 and G1 are significantly different.

The rpoD gene coding for the primary transcription sigma factor, sigma70, and its entire operon were cloned from strain mt-2 of the purple soil bacterium Pseudomonas putida. Comparison of the deduced amino acid sequence of Ppmt-2 sigma70 with that of sigma70 from P. putida strain G1 shows that the two proteins differ in their primary structure, molecular weight, and isoelectric point. The significance of this difference is discussed in terms of bacterial taxonomy and transcription regulation.

Amino Acid Sequence↗
PubMed1997

Movement of Bax from the cytosol to mitochondria during apoptosis.

Bax, a member of the Bcl-2 protein family, accelerates apoptosis by an unknown mechanism. Bax has been recently reported to be an integral membrane protein associated with organelles or bound to organelles by Bcl-2 or a soluble protein found in the cytosol. To explore Bcl-2 family member localization in living cells, the green fluorescent protein (GFP) was fused to the NH2 termini of Bax, Bcl-2, and Bcl-XL. Confocal microscopy performed on living Cos-7 kidney epithelial cells and L929 fibroblasts revealed that GFP-Bcl-2 and GFP-Bcl-XL had a punctate distribution and colocalized with a mitochondrial marker, whereas GFP-Bax was found diffusely throughout the cytosol. Photobleaching analysis confirmed that GFP-Bax is a soluble protein, in contrast to organelle-bound GFP-Bcl-2. The diffuse localization of GFP-Bax did not change with coexpression of high levels of Bcl-2 or Bcl-XL. However, upon induction of apoptosis, GFP-Bax moved intracellularly to a punctate distribution that partially colocalized with mitochondria. Once initiated, this Bax movement was complete within 30 min, before cellular shrinkage or nuclear condensation. Removal of a COOH-terminal hydrophobic domain from GFP-Bax inhibited redistribution during apoptosis and inhibited the death-promoting activity of both Bax and GFP-Bax. These results demonstrate that in cells undergoing apoptosis, an early, dramatic change occurs in the intracellular localization of Bax, and this redistribution of soluble Bax to organelles appears important for Bax to promote cell death.

Animals↗
PubMed1997

Nuclear membrane dynamics and reassembly in living cells: targeting of an inner nuclear membrane protein in interphase and mitosis.

The mechanisms of localization and retention of membrane proteins in the inner nuclear membrane and the fate of this membrane system during mitosis were studied in living cells using the inner nuclear membrane protein, lamin B receptor, fused to green fluorescent protein (LBR-GFP). Photobleaching techniques revealed the majority of LBR-GFP to be completely immobilized in the nuclear envelope (NE) of interphase cells, suggesting a tight binding to heterochromatin and/or lamins. A subpopulation of LBR-GFP within ER membranes, by contrast, was entirely mobile and diffused rapidly and freely (D = 0. 41 +/- 0.1 microm2/s). High resolution confocal time-lapse imaging in mitotic cells revealed LBR-GFP redistributing into the interconnected ER membrane system in prometaphase, exhibiting the same high mobility and diffusion constant as observed in interphase ER membranes. LBR-GFP rapidly diffused across the cell within the membrane network defined by the ER, suggesting the integrity of the ER was maintained in mitosis, with little or no fragmentation and vesiculation. At the end of mitosis, nuclear membrane reformation coincided with immobilization of LBR-GFP in ER elements at contact sites with chromatin. LBR-GFP-containing ER membranes then wrapped around chromatin over the course of 2-3 min, quickly and efficiently compartmentalizing nuclear material. Expansion of the NE followed over the course of 30-80 min. Thus, selective changes in lateral mobility of LBR-GFP within the ER/NE membrane system form the basis for its localization to the inner nuclear membrane during interphase. Such changes, rather than vesiculation mechanisms, also underlie the redistribution of this molecule during NE disassembly and reformation in mitosis.

Animals↗
PubMed1997

Substrate DNA and cofactor regulate the activities of a multi-functional restriction-modification enzyme, BcgI.

The BcgI restriction-modification system consists of two subunits, A and B. It is a bifunctional protein complex which can cleave or methylate DNA. The regulation of these competing activities is determined by the DNA substrates and cofactors. BcgI is an active endonuclease and a poor methyltransferase on unmodified DNA substrates. In contrast, BcgI is an active methyltransferase and an inactive endonuclease on hemimethylated DNA substrates. The cleavage and methylation reactions share cofactors. While BcgI requires Mg2+and S -adenosyl methionine (AdoMet) for DNA cleavage, its methylation reaction requires only AdoMet and yet is significantly stimulated by Mg2+. Site-directed mutagenesis was carried out to investigate the relationship between AdoMet binding and BcgI DNA cleavage/methylation activities. Most substitutions of conserved residues forming the AdoMet binding pocket in the A subunit abolished both methylation and cleavage activities, indicating that AdoMet binding is an early common step required for both cleavage and methylation. However, one mutation (Y439A) abolished only the methylation activity, not the DNA cleavage activity. This mutant protein was purified and its methylation, cleavage and AdoMet binding activities were tested in vitro . BcgI-Y439A had no detectable methylation activity, but it retained 40% of the AdoMet binding and DNA cleavage activities.

Base Sequence↗
PubMed1997

Cloning and analysis of the dnaG gene encoding Pseudomonas putida DNA primase.

The dnaG gene coding for primase, a key enzyme in DNA replication, has been isolated from chromosomal DNA of the soil bacterium Pseudomonas putida. It maps within the putative MMS operon, between the rpsU and rpoD genes. Comparison of the deduced amino acid sequence of P. putida DnaG with sequences of other known bacterial primases reveals the presence of a possible regulatory region which would be unique to pseudomonads. The analysis of nucleotide sequence suggests that stable folding of the dnaG mRNA may significantly contribute to the low level of its expression within a cell.

Amino Acid Sequence↗
PubMed1997

Engineering subunit association of multisubunit proteins: a dimeric streptavidin.

A dimeric streptavidin has been designed by molecular modeling using effective binding free energy calculations that decompose the binding free energy into electrostatic, desolvation, and side chain entropy loss terms. A histidine-127 --> aspartic acid (H127D) mutation was sufficient to introduce electrostatic repulsion between subunits that prevents the formation of the natural tetramer. However, the high hydrophobicity of the dimer-dimer interface, which would be exposed to solvent in a dimeric streptavidin, suggests that the resulting molecule would have very low solubility in aqueous media. In agreement with the calculations, a streptavidin containing the H127D mutation formed insoluble aggregates. Thus, the major design goal was to reduce the hydrophobicity of the dimer-dimer interface while maintaining the fundamental structure. Free energy calculations suggested that the hydrophobicity of the dimer-dimer interface could be reduced significantly by deleting a loop from G113 through W120 that should have no apparent contact with biotin in a dimeric molecule. The resulting protein, containing both the H127D mutation and the loop deletion, formed a soluble dimeric streptavidin in the presence of biotin.

Aspartic Acid↗
PubMed1997

Differential activity of progesterone and glucocorticoid receptors on mouse mammary tumor virus templates differing in chromatin structure.

In vivo, transcription factors interact with promoters having complex nucleoprotein structures. The transiently expressed progesterone receptor (PR) efficiently activates a transfected mouse mammary tumor virus (MMTV) promoter but is a poor activator of the MMTV promoter when it acquires an ordered chromatin structure as an endogenous, replicating gene. We show that the deficiency in PR activity is not due to insufficient expression of either B or A isoforms or competition between the two types of MMTV templates. Rather, this deficiency reflects an inability to induce the chromatin remodeling event that is required for activation of the replicated MMTV template. To determine whether this characteristic is common to transiently expressed steroid receptors or specific to the PR, we examined the activity of transiently expressed glucocorticoid (GR) receptor. Unlike the PR, the transiently expressed GR is an effective activator of both MMTV templates and efficiently induces the necessary chromatin remodeling event at the replicated template. These results indicate that the GR and PR have unique requirements for activation of promoters with ordered chromatin structure. These differences may provide a mechanism for establishing target gene specificity in vivo for steroid receptors that recognize and bind to identical DNA sequences.

Animals↗
PubMed1997

Differential display of genome subsets containing specific interspersed repeats.

A genomic differential display method was developed that analyzes many restriction fragment length polymorphisms simultaneously. Interspersed repeat sequences were used to reduce DNA sample complexity and to target genomic subsets of interest. This work focused on trinucleotide repeats because of their importance in human inherited diseases. Immobilized repeat-containing oligonucleotides were used to capture genomic DNA fragments containing sequences complementary to the oligonucleotide. Captured fragments were amplified by PCR and fluorescently labeled using primers complementary to the repeat sequence and/or to the known sequences ligated to the ends of the restriction fragments. The labeled PCR fragments were displayed by size on a high-resolution automated fluorescent DNA sequencing instrument. Although there was a conservation in the overall pattern of displayed genome subsets, many clear and reproducible differences were detected when genomes from different individuals were compared. Fewer differences were detected within, than between, monozygotic twin pair genomes. In control experiments, the method distinguished between Huntington disease alleles with normal and expanded CAG repeat lengths.

Base Sequence↗
PubMed1997

Aldosterone and dexamethasone stimulate calcineurin activity through a transcription-independent mechanism involving steroid receptor-associated heat shock proteins.

Heat shock proteins (HSP) are components of the steroid receptor complex and are released into the cell cytosol after hormone binding. We tested whether HSPs released from steroid receptors mediate an increase in calcineurin phosphatase activity by steroid hormones. Aldosterone increased calcineurin activity in microdissected rat cortical collecting ducts (CCD) and connecting tubules, but not in proximal tubules, medullary thick ascending limb, or outer medullary collecting ducts. In contrast, 5 microM dexamethasone increased calcineurin activity in both CCD and proximal tubules. Aldosterone increased CCD calcineurin activity after 30 min and this response was blocked by spironolactone, but not by actinomycin D. An antibody recognizing HSP-56 did not change basal calcineurin activity, but completely blocked the stimulation of calcineurin by aldosterone. Rapamycin, an immunosuppressive drug that stabilizes the HSP-steroid receptor complex, also blocked the aldosterone response, whereas HSP-90 or HSP-70 increased calcineurin activity in permeabilized CCD. In summary, (a) aldosterone increases calcineurin activity in CCD through a transcription-independent process; (b) maneuvers inactivating HSP-56 or slowing HSP disassociation from the receptor complex blocks stimulation of calcineurin by steroid hormones; (c) HSP-90 and HSP-70 increase CCD calcineurin activity in the absence of steroid hormone. We conclude that HSPs released from transformed steroid receptors can stimulate calcineurin activity through a transcription-independent pathway.

Adrenalectomy↗