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Biomedical subjects

L J Smith

Publications and source records attributed to L J Smith.

At least 91 records · Page 5Linked to original sources
PubMed1995

c-myc antisense oligodeoxyribonucleotides inhibit proliferation of non-small cell lung cancer.

BACKGROUND: Mutation or deregulation of certain cellular genes (protooncogenes) results in expression of proteins that appear to promote malignant transformation. Human non-small cell lung cancer has been documented to express many such oncogenes including c-myc, bcl-2, and mutant p53. Antisense oligodeoxyribonucleotides (ASODN) complementary to these oncogenes were tested on three non-small cell lung cancer cell lines for their efficacy in inhibiting cellular proliferation and oncoprotein expression. METHODS: Established non-small cell lung cancer cell lines A427, SKMES-1, and A549 were grown in the presence of ASODNs complementary to messenger RNA of c-myc, bcl-2, p53, or controls at 1 mumol/L or 10 mumol/L concentrations for 4 or 10 days. Cellular proliferation was measured by tritiated thymidine uptake. Flow cytometry was used to quantitate oncoprotein expression. Intranuclear ASODN uptake was documented by fluoresceine-tagged ASODNs. RESULTS: Fluoresceine-tagged ASODNs were readily taken up by all cell lines. c-myc, as well as bcl-2 and p53 ASODNs, were found to inhibit proliferation of all cell lines significantly compared with controls, most notably in line A549 (40.1% +/- 7.1% of control, p = 0.000 with c-myc ASODN). Antisense c-myc reduced c-myc protein by as much as 71.3% in A427, although protein levels were only minimally reduced in the viable cells of the other lines. CONCLUSIONS: c-myc ASODNs inhibit proliferation of non-small cell lung cancer cell lines as well as reduce c-myc protein expression. Antisense bcl-2 and p53 also cause similar growth inhibition. These results suggest a critical role for activation of these oncogenes in the growth of cultured lung cancer cells. Furthermore, the efficacy and rapid cellular uptake of ASODNs support the potential role of antisense targeting of oncogene expression for pharmacologic control of non-small cell lung cancer.

Carcinoma, Non-Small-Cell Lung↗
PubMed1995

Differential regulation of leukotriene and platelet-activating factor synthesis in rat alveolar macrophages.

Leukotrienes (LT) and platelet-activating factor (PAF) are synthesized by several lung cells, including alveolar macrophages (AM), and may contribute to the airway inflammation that characterizes asthma. Phospholipases A2 (PLA2) can release arachidonic acid and lysophosphatidylcholine (lysoPC), precursors for leukotriene and PAF synthesis, respectively, from membrane phospholipids. The present study sought to determine the extent to which this common initial step contributes to leukotriene and PAF synthesis in rat AM. AM were obtained by lung lavage, cultured, and exposed to one or more of the following: PAF, zileuton (5-lipoxygenase inhibitor), Ro 25-4331 (dual PLA2 inhibitor), and either A23187 (Cal) or zymosan. The following measurements were made: LTB4 synthesis, PAF synthesis, and PLA2 activity. CaI stimulation increased PAF synthesis 3-fold (P < 0.001), LTB4 synthesis 20-fold (P < 0.001), and PLA2 activity 52% (P < 0.001). Incubation with PAF (2.5 microM) for 10 min decreased basal LTB4 synthesis 33% (P < 0.001), but it had no effect on basal PAF synthesis or PLA2 activity. The same dose of PAF (2.5 microM) decreased CaI-stimulated PAF synthesis 40% (P < 0.02). After CaI stimulation of PAF-pretreated cells, a relationship was found between PAF-induced changes in PLA2 activity and LTB4 synthesis (r = 0.68; P < 0.001) but not between changes in PLA2 activity and PAF synthesis. Zileuton (1 microM) decreased basal and CaI-stimulated LTB4 synthesis 50% (P < 0.02), and 80% (P < 0.002) respectively, but it did not alter PAF synthesis.

Animals↗
PubMed1995

Intratracheal instillation of keratinocyte growth factor decreases hyperoxia-induced mortality in rats.

Alveolar type II cell proliferation occurs after many forms of lung injury and is thought to play a critical role in alveolar epithelial repair. Keratinocyte growth factor/fibroblast growth factor 7 (KGF) has been shown to promote alveolar type II cell growth in primary culture and alveolar epithelial hyperplasia in vivo. In this study, we used immunohistochemical analysis to determine the intrapulmonary distribution and cellular localization of recombinant human KGF (rhKGF) instilled into the trachea of rats. 6 h after administration, immunoreactive KGF was observed within the lung parenchyma and along alveolar epithelial cell membranes. By 18-24 h, KGF was detected intracellularly in alveolar epithelial cells and intraalveolar macrophages. Immunoreactive KGF was not demonstrable 48 h after delivery or in lung sections from PBS-treated animals. Intratracheal instillation of 5 mg/kg rhKGF stimulated a marked, time-dependent increase in the alveolar type II cell specific labeling index to a maximum level of 33 +/- 3% 48 h after rhKGF administration compared with 1.3 +/- 0.3% after PBS instillation. In addition, this increase in type II cell proliferation in vivo was documented by flow cytometric analysis of isolated type II cells which revealed a nearly fivefold increase in the proportion of cells traversing through the S and G2/M phases of the cell cycle. To test the hypothesis that KGFs effects on type II cells in vivo might affect the response to lung injury, rats were treated with rhKGF and exposed to hyperoxia. Animals that received 1 or 5 mg/kg rhKGF exhibited dramatically reduced mortality (P < 0.001, for both doses). Survival for animals treated with 0.1 mg/kg rhKGF was not significantly different from either untreated rats or animals treated with heat-denatured rhKGF. The lungs of rhKGF-treated animals that survived hyperoxia exposure had minimal hemorrhage and no exudate within the intraalveolar space. These experiments established that intratracheal administration of rhKGF stimulated alveolar type II cell proliferation in vivo and reduced hyperoxia-induced lung injury in rats. Directed delivery of KGF to the lungs may provide a therapeutic strategy to preserve or restore the alveolar epithelium during exposure to hyperoxia or other injurious agents.

Animals↗
PubMed1995

A score sheet for evaluating breastfeeding educational materials.

Lactation consultants and other professionals are frequently asked to review or recommend a video, pamphlet, or teaching aid for a specific audience. This article lists positive features that support the art and science of breastfeeding, and identifies errors and concepts detrimental to the course of breastfeeding. Six central messages are detailed: milk supply, comfort during breastfeeding, health of baby and mother, social support, presentation of the message, and packaging, including the financial motivation of the author.

Breast Feeding↗
PubMed1995

Neutrophil activation in paediatric extracorporeal circuits: effect of circulation and temperature variation.

OBJECTIVE: Upregulation of neutrophil adhesion molecules (CD11b and L-selectin) and release of a modulating cytokine (IL8) have been reported in vivo and in vitro in adult cardiopulmonary bypass. The aim of this study was to determine whether paediatric bypass preparations have similar influences and whether neutrophil-endothelium interactions are required for IL8 release. METHODS: In vitro paediatric cardiopulmonary bypass circuits (n = 15) were constructed (identical to those used clinically), as well as static loops (n = 15) using donor blood. The effects of circulation and temperature (17 degrees C, 25 degrees C, 37 degrees C) on the initiation of acute inflammation were examined. Cellular expressions of neutrophil adhesion molecules CD11b and L-selectin were assayed by immunofluorescence technique, and serum IL8, IL6, TNF-alpha, leucocyte elastase, and terminal complement complex were measured by ELISA. RESULTS: In all experiments, an immediate increase in CD11b expression occurred [median values, in relative fluorescence units: 64.9 (range 45.3-212.9) at rest; 365.2 (205-835.4) at 10 min; P < 0.001], along with a decrease in L-selectin expression [153.5 (115.5-220.7) at rest; 42 (12-134) at 10 min; P < 0.01]. Serum concentrations of the following increased gradually and were higher in circulation than in static loops: IL8 [1500 (500-2500) pg.ml-1 in circuit v 600 (180-1500) pg.ml-1 in loop, P < 0.001]; TNF-alpha P < 0.05]; and terminal complement complex [25.9 (6.8-120) v 4.7 (0-21.6) AU.ml-1, P < 0.01]. Cooling decreased and rewarming increased upregulation of CD11b and downregulation of L-selectin and release of IL8. IL6 was undetectable. CONCLUSIONS: In the absence of endothelium, in vitro paediatric cardiopulmonary bypass causes profound acute inflammatory changes in donor blood with release of IL8. These changes were greater than in adult cardiopulmonary bypass. Temperature variation and circulation modulate the responses.

Adult↗
PubMed1995

Diagnosis and treatment of allergic rhinitis.

Allergic rhinitis is frequently seen by primary care providers. Symptoms include rhinorrhea, itching of nose and eyes, nasal congestion, and sneezing. They occur when the patient is exposed to antigens stimulating mediator release. History is essential to assist differential diagnosis and provide education. This article reviews common findings of the physical exam, diagnostic testing, and differential diagnosis. The first line of treatment is avoidance, followed by medications such as antihistamines, decongestants, and nasal steroids. If these treatments are not effective, then referral to an allergist is necessary for further workup and possible treatment with immunotherapy.

Diagnosis, Differential↗
PubMed1994

Thrombin generation in patients undergoing carotid endarterectomy: implications in acute vessel wall closure and antithrombotic therapy.

We determined the extent and duration of activation of coagulation during the first 24 hours after carotid endarterectomy. Serial blood samples were collected before, during and after surgery from 10 patients with severe stenosis (> 70%) undergoing carotid endarterectomy with heparinization. Platelet poor plasmas prepared from these samples, were analysed for activated prothrombin fragment F1 + 2 and thrombin-antithrombin III (TAT) levels, using commercially available ELISA kits. F1 + 2 and TATs were measured as indices of thrombin generation and inhibition respectively. Baseline F1 + 2 and TAT levels were 1.19 +/- 0.27 nMol/ml and 17 +/- 10 pMol/ml, respectively. Neither the F1 + 2 nor TAT level increased during surgery at a time when the patients were heparinized. However, both the F1 + 2 and TAT levels increased significantly within 3 hours after surgery and after the heparin had been neutralized with protamine, (p < 0.01). Moreover, both the F1 + 2 and TAT levels remained elevated in 75% of the patients for at least 24 hours, p < 0.01. We conclude that i) thrombin generation is significant post surgery in patients undergoing carotid endarterectomy despite their receiving heparin during surgery; ii) heparin may not be the ideal anticoagulant for carotid endarterectomy; and iii) persistent thrombin generation may contribute to early post-endarterectomy ischemic events.

Acute Disease↗
PubMed1994

Solution structure of a peptide fragment of human alpha-lactalbumin in trifluoroethanol: a model for local structure in the molten globule.

BACKGROUND: At low pH, human alpha-lactalbumin forms a partly folded molten globule state that contains a non-native clustering of the side chains of Tyr103, Trp104 and His107. In order to understand the conformation of this region of the protein in the molten globule state, we investigated the structure of a peptide corresponding to residues 101-110 of human alpha-lactalbumin in trifluoroethanol. RESULTS: We determined the structure of the 101-110 peptide from an NMR data set of 145 nuclear Overhauser effects and nine 3JHN alpha coupling constants, using an ensemble calculation approach to take into account the possibilities of conformational averaging of the data. The backbone of residues 3-10 in the peptide adopts a series of turns, that involving residues 5-8 being the best defined, while the side chains of residues 1, 3, 4, 5, 6 and 7 form a hydrophobic cluster. CONCLUSIONS: The peptide conformation differs from that previously determined for residues 101-110 in crystal structures of native alpha-lactalbumin determined at both high and low pH, particularly in the relative orientations of the side chains. The series of turns seen in the peptide could, however, be related to the alpha-helical structure seen for residues 104-111 in crystals at high pH, and may be important in the molten globule state for bringing the peptide chain into a compact conformation where favourable interactions between the side chains can occur.

Amino Acid Sequence↗
PubMed1994

Prevention of thrombus formation and growth by antithrombin III and heparin cofactor II-dependent thrombin inhibitors: importance of heparin cofactor II.

Heparin (HEP) prevents thrombus formation (TF) and thrombus growth (TG), by accelerating thrombin (THR) inhibition by antithrombin III (ATIII). Recent studies suggest that dermatan sulphate which catalyzes thrombin inhibition by heparin cofactor II (HCII), can inhibit TF and TG as effectively as HEP. This study compared the antithrombotic effects of HEP and another agent, Sulodexide (SLX) which catalyzes thrombin inhibition by ATIII and HCII simultaneously. TF was induced in rabbit jugular veins, using the stasis/hypercoagulation model. TG was measured as the accretion of 125I-fibrin onto existing thrombi in rabbit jugular veins. HEP and SLX inhibited TF when given in doses of 10 and 5 anti-thrombin U/kg, respectively. SLX (16 anti-thrombin U/kg or 260 micrograms/kg) was more effective than HEP (120 anti-thrombin U/kg or 800 micrograms/kg) in preventing TG when administered either as a bolus or by continuous infusion. These data suggest that agents which accelerate THR inhibition by both ATIII and HCII simultaneously, can inhibit TF and TG with less systemic anticoagulation than comparable antithrombotic doses of HEP.

Animals↗
PubMed1994

Analysis of the solution structure of human interleukin-4 determined by heteronuclear three-dimensional nuclear magnetic resonance techniques.

Human interleukin-4 (IL-4) is a member of the family of haemopoietic cytokines that modulate cell proliferation and differentiation within the immune system. It has a four-helix-bundle structure, and possesses a high degree of mobility in certain regions, notably in the two long loops running the length of the bundle in its up-up-down-down topology. Information from a variety of three-dimensional heteronuclear NMR experiments, including chemical shifts, coupling constants and NOE data, is analysed in terms of the solution structure of IL-4. In addition, structure calculations with and without specific restraints such as hydrogen bond location or torsion angle restrictions are compared in the light of the dynamic behaviour of the polypeptide chain. Particular emphasis is placed on defining the lengths and positions of secondary structure elements, and on the likely structural preferences within the less well ordered loop regions. The overall topology of IL-4 is compared with those defined in recent structure determinations of related proteins. This analysis is combined with recent mutagenesis data to propose a possible mode of interaction of IL-4 with its receptor.

Crystallography, X-Ray↗
PubMed1994

Sequence of a cDNA encoding the p53 protein in rhesus monkey (Macaca mulatta).

A 2184-nucleotide (nt) sequence of the p53 gene in Rhesus monkey (Macaca mulatta) was determined in order to facilitate the use of the Rhesus as an animal model in the testing of novel antisense oligodeoxyribonucleotides for a variety of human cancers, including acute myelogenous leukemia (AML). Within overlapping regions, we found greater than 95% identity between the Rhesus and human p53 sequences, and greater than 98% identity between Rhesus and African green monkey p53 sequences. The deduced amino acid (aa) sequence of the p53 protein is highly conserved between human and Rhesus monkey, with only 18 minor differences in 393 aa.

Amino Acid Sequence↗
PubMed1994

A behavioral approach to the treatment of nonretentive nocturnal encopresis in an adult with severe learning disabilities.

Encopresis is widely assumed to be "normal" among individuals who are learning disabled. Yet, reports on its prevalence, etiology and treatment are rare and mainly confined to secondary encopresis or to populations with milder learning disabilities. The behavioral treatment of a life-long nonretentive, nocturnal encopresis is described for a young adult with severe learning and physical disabilities living at home. Treatment involved waking prior to soiling, time-limited use of suppositories and rewards for appropriate evacuation. Soiling decreased to zero over 20 weeks. Time of waking was then gradually "shaped" back to normal.

Adult↗
PubMed1994

Mite allergen control with acaricide fails.

BACKGROUND: We compared the effects of an acaricide, benzyl benzoate, with the effects of baking soda control applied to bedroom and living room carpets on house dust mite allergen levels, lung function, and medication use in 12 adult patients with asthma for 12 months. METHODS: This was a randomized, double-blind, placebo-controlled study. Patients were enrolled from the allergy clinic of a large tertiary care center in a metropolitan area. All patients had positive dust mite puncture test results. Six patients used benzyl benzoate, and six used baking soda. Other aggressive mite control measures were implemented uniformly in each group. Subjects were to make two carpet applications, at baseline and at 6 months according to the manufacturer's recommendations. Dust samples were collected in bedroom and living room carpets at 0, 3, 6, 9, and 12 months; and quantities of Der p I and Der f I allergens were determined. Spirometry was done every 3 months, and peak flow rates were recorded for 10 days after each dust sampling. RESULTS: There were no significant differences in mean allergen levels between the two groups over time at either site. There were no significant changes in lung function or medication use for either group. CONCLUSIONS: Benzyl benzoate powder applications may not be effective when done according to manufacturer's instructions. Further studies are necessary to test effectiveness when applied more frequently and for longer periods.

Adult↗
PubMed1994

Familial enteropathy with villous edema and immunoglobulin G2 subclass deficiency.

We describe a familial form of recurrent acute, life-threatening secretory diarrhea associated with distinctive jejunal histologic changes and IgG2 subclass deficiency. Symptoms begin abruptly with anorexia and vomiting, and progress within hours to massive secretory diarrhea and shock with profound neutropenia and hypoproteinemia, including hypoalbuminemia and hypogammaglobulinemia. Affected survivors recover quickly and thereafter grow and develop normally. Biopsy specimens obtained during remission from 3 adults and 11 children show club-shaped jejunal villi broadened by edema and histiocytes with imbibed fluid; the overlying intestinal epithelium and brush border appear normal, but the basement membrane is interrupted in some areas. No characteristic microorganisms have been identified in association with the syndrome. Clinical manifestations cease in the second decade, but the abnormal jejunal histologic pattern persists into adult life. Female and male patients are equally affected, although all fatal cases have been in female subjects. Inheritance appears dominant with variable penetrance: one family member without a history of diarrhea has characteristic biopsy findings and another appears to be an obligate carrier with normal biopsy findings. Affected individuals have a reduced serum concentration of IgG2. We believe that this familial enteropathy is a unique entity, not previously described.

Adolescent↗
PubMed1994

The paradox of the renin-angiotensin system in chronic renal disease.

Despite normal to suppressed levels of renin activity in chronic renal disease, multiple lines of evidence suggest a role for the RAS, especially its intrarenal expression, in several critical aspects of this condition. Alterations in the distribution and control of components of the renal RAS could account for localized areas of activation of this system. Renal scarring may be particularly important as a major stimulus to renin synthesis in the diseased kidney. While both intrarenal and systemic hypertension may depend in part upon actions of the RAS, other non-hemodynamic actions of the RAS may also contribute to the adaptation of residual nephrons as well as their progressive injury.

Animals↗
PubMed1994

Comparison of four independently determined structures of human recombinant interleukin-4.

Four independent structures of human interleukin-4, two determined by nuclear magnetic resonance techniques and two by X-ray diffraction, have been compared in detail. The core of this four helix bundle protein is very similar in all the structures but there are some differences in loop regions that are known to be mobile in solution. Careful comparison of the experimental data sets and the methods of analysis of the different laboratories has provided clues to the sources of most of the differences, and also answered some general questions about the accuracy of protein structure determination by these two techniques.

Crystallography, X-Ray↗