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Biomedical subjects

K Lund

Publications and source records attributed to K Lund.

At least 19 recordsLinked to original sources
PubMed1999

Foreign gene expression in Corynebacterium pseudotuberculosis: development of a live vaccine vector.

A defined phospholipase D mutant of Corynebacterium pseudotuberculosis, designated Toxminus, was used as a live vector to express and deliver a range of candidate vaccine antigens to sheep. Expression levels of the foreign genes in Toxminus were evaluated when directed from a number of different promoters, both constitutively expressed and inducible, as fusions with expressed genes including a signal sequence, and from chromosomal and episomal loci. In general expression levels were low and it appeared that some of the recombinant proteins were tolerated by C. pseudotuberculosis Toxminus better than others. Gene expression was however sufficiently high for three of the genes to elicit antibody responses specific to the recombinant protein following a single dose of the live Toxminus vector vaccine. This work suggests that C. pseudotuberculosis Toxminus has potential for development as a live veterinary vaccine vector.

Animals↗
PubMed1999

Synthesis of beta-tubulin, actin, and other proteins in axons of sympathetic neurons in compartmented cultures.

The proteins needed for growth and maintenance of the axon are generally believed to be synthesized in the cell bodies and delivered to the axons by anterograde transport. However, recent reports suggest that some proteins can also be synthesized within axons. We used [35S]methionine metabolic labeling to investigate axonal protein synthesis in compartmented cultures of sympathetic neurons from newborn rats. Incubation of distal axons for 4 hr with [35S]methionine resulted in a highly specific pattern of labeled axonal proteins on SDS-PAGE, with 4 prominent bands in the 43-55 kDa range. The labeled proteins in axons were not synthesized in the cell bodies, because they were also produced by axons after the cell bodies had been removed. Two of the proteins were identified by immunoprecipitation as actin and beta-tubulin. Axons synthesized <1% of the actin and tubulin synthesized in the cell bodies and transported into the axons, and 75-85% inhibition of axonal protein synthesis by cycloheximide and puromycin failed to inhibit axonal elongation. Nonetheless, the specific production by axons of the major proteins of the axonal cytoskeleton suggests that axonal protein synthesis arises from specific mechanisms and likely has biological significance. One hypothetical scenario involves neurons with long axons in vivo in which losses from turnover during axonal transport may limit the availability of cell body synthesized proteins to the distal axons. In this case, a significant fraction of axonal proteins might be supplied by axonal synthesis, which could, therefore, play important roles in axonal maintenance, regeneration, and sprouting.

Actins↗
PubMed1999

Fluorescence In situ hybridization assay using peptide nucleic acid probes for differentiation between tuberculous and nontuberculous mycobacterium species in smears of mycobacterium cultures.

TB PNA FISH is a new fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for differentiation between species of the Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) in acid-fast bacillus-positive (AFB+) cultures is described. The test is based on fluorescein-labelled PNA probes that target the rRNA of MTC or NTM species applied to smears of AFB+ cultures for microscopic examination. Parallel testing with the two probes serves as an internal control for each sample such that a valid test result is based on one positive and one negative reaction. TB PNA FISH was evaluated with 30 AFB+ cultures from Denmark and 42 AFB+ cultures from Thailand. The MTC-specific PNA probe showed diagnostic sensitivities of 84 and 97%, respectively, and a diagnostic specificity of 100% in both studies, whereas the NTM-specific PNA probe showed diagnostic sensitivities of 91 and 64%, respectively, and a diagnostic specificity of 100% in both studies. The low sensitivity of the NTM-specific PNA probe in the Thai study was due to a relatively high prevalence of Mycobacterium fortuitum, which is not identified by the probe. In total, 63 (87%) of the cultures were correctly identified as MTC (n = 46) or NTM (n = 17), whereas the remaining 9 were negative with both probes and thus the results were inconclusive. None of the samples were incorrectly identified as MTC or NTM; thus, the predictive value of a valid test result obtained with TB PNA FISH was 100%.

Base Sequence↗
PubMed1999

Imageability and distributional typicality measures of nouns and verbs in contemporary English.

Dissociations between noun and verb processing are not uncommon after brain injury; yet, precise psycholinguistic comparisons of nouns and verbs are hampered by the underrepresentation of verbs in published semantic word norms and by the absence of contemporary estimates for part-of-speech usage. We report herein imageability ratings and rating response times (RTs) for 1,197 words previously categorized as pure nouns, pure verbs, or words of balanced noun-verb usage on the basis of the Francis and Kucera (1982) norms. Nouns and verbs differed in rated imageability, and there was a stronger correspondence between imageability rating and RT for nouns than for verbs. For all word types, the image-rating-RT function implied that subjects employed an image generation process to assign ratings. We also report a new measure of noun-verb typicality that used the Hyperspace Analog to Language (HAL; Lund & Burgess, 1996) context vectors (derived from a large sample of Usenet text) to compute the mean context distance between each word and all of the pure nouns and pure verbs. For a subset of the items, the resulting HAL noun-verb difference score was compared with part-of-speech usage in a representative sample of the Usenet corpus. It is concluded that this score can be used to estimate the extent to which a given word occurs in typical noun or verb sentence contexts in informal contemporary English discourse. The item statistics given in Appendix B will enable experimenters to select representative examples of nouns and verbs or to compare typical with atypical nouns (or verbs), while holding constant or covarying rated imageability.

Adult↗
PubMed1999

Increased CD3 positive cells in bronchoalveolar lavage fluid after hydrogen fluoride inhalation.

OBJECTIVES: This study examined whether experimental hydrogen fluoride exposure for 1 hour induces an inflammatory response in the lower respiratory tract that is detectable in bronchoalveolar lavage fluid. METHODS: Nineteen healthy, nonsmoking men were exposed for 1 hour to constant low (<0.6 mg/m3), intermediate (0.7-2.4 mg/m3), or high (2.5-5.2 mg/m3) concentrations of hydrogen fluoride. Bronchoalveolar lavage was performed at least 3 weeks before and 24 hours after the exposure. For 15 subjects differential countings were performed. RESULTS: There was a significant increase in the percentage of CD3 positive cells in the bronchial portion for those exposed to "intermediate" and "high" concentrations. For the "high" exposure group the increase in the bronchoalveolar portion was also significant. A significant correlation was found between the increase in the percentage of lymphocytes and CD3 positive cells in the bronchoalveolar portion (Spearman's coefficient r=0.68, P=0.008). Myeloperoxidase and interleukin-6 increased significantly in the bronchial portion for those exposed to "high" concentrations. There was a significant increase in myeloperoxidase (P=0.005) for all the exposures, while there was a decrease in E-selectin (P=0.007). CONCLUSIONS: Hydrogen fluoride may induce an inflammatory reaction in the airways at concentrations that can occur in the ambient air in the primary aluminum industry.

Adult↗
PubMed1999

Direct detection and identification of Mycobacterium tuberculosis in smear-positive sputum samples by fluorescence in situ hybridization (FISH) using peptide nucleic acid (PNA) probes.

SETTING: Peptidenucleic acid (PNA) probesdesigned for specific detection of mycobacteria of the Mycobacterium tuberculosis complex (MTC) and other non-tuberculous mycobacterium species (NTM) are shown to be able to penetrate the mycobacterial cell wall and subsequently hybridize in situ to complementary rRNA. OBJECTIVE: To demonstrate the use of fluorescein-labelled PNA probes for detection and identification of M. tuberculosis in smear-positive sputum samples. DESIGN: The sensitivity and specificity of the PNA probes were investigated by fluorescence in situ hybridization (FISH) using cultures of mycobacterium strains representing species of the MTC and NTM, respectively. RESULTS: M. tuberculosis strains were detected by FISH using specific fluorescein-labelled PNA probes directly in smear-positive sputum samples without changing the morphology of the cells. CONCLUSION: PNA probes allow for rapid diagnosis of tuberculosis in smear-positive cases.

Humans↗
PubMed1998

Overlapping 3'-end formation signals and ARS elements: tightly linked but functionally separable.

3'-End formation signals are closely associated with autonomous replicating sequences (ARSs) in Saccharomyces cerevisiae in that ARSs frequently contain signals that direct 3'-end formation (Chen et al., 1996). Mutationally-inactivated ARSs that co-reside with 3'-end formation sequences do not disrupt 3'-end formation, thus demonstrating that replication function does not affect termination function. To test the corollary possibility that 3'-end formation is important for replication function, we made point mutations in ARS305 that increase readthrough of the 3'-end formation signals and determined plasmid replication efficiency. Replication efficiency, as assessed by plasmid stability assays, was not altered by mutations affecting 3'-end formation when transcription through the ARS was either absent or highly-induced. Under conditions of high-level transcription through the ARS, the rate of plasmid loss in both wild-type and mutated terminators increased over five-fold from rates observed during transcriptionally repressed conditions. This result indicates that the native 3'-end formation signal is incapable of protecting the replication function when high levels of transcription are directed into the ARS. Thus, the compact nature of the S. cerevisiae genome, rather than a functional inter-dependence, may account for close association of transcription terminators and ARSs.

Base Sequence↗
PubMed1998

Recovery of beat-to-beat variations of QRS.

There is a growing interest in the analysis of beat-to-beat variations of the morphology (BBM) of cardiac waves in electrocardiograms (ECG). Such analyses are confronted with the low BBM-to-noise ratio. An ECG clustering technique is introduced that brings the benefits of signal averaging to BBM analysis and recovers the beat-to-beat pattern of BBM. ECG clustering aligns waves and sorts them into clusters. The precision of the alignment was enhanced by sub-sample alignment. Kohonen's self-organising neural networks identified the clusters of the cardiac waves during training. The subsequent clustering of a wave results in a label for the closest cluster, a distance to the cluster and optimal alignment. Furthermore, ECG clustering avoids base-line variations and amplitude modulation sufficiently to be applied to the QRS wave in the raw ECG. The technique is demonstrated on 14 subjects with coronary heart disease and no myocardial infarction, myocardial infarction, or inducible ventricular tachycardia. ECG clustering is a general-purpose technique for beat-to-beat analysis, where the variations are cyclic as in the sinus rhythm. Results show that beat-to-beat variations in the QRS morphology are in general cyclic, with a main period of about four cardiac cycles. All calculations were performed with the Cardio software.

Coronary Disease↗
PubMed1998

Renal effects of losartan and amlodipine in hypertensive patients with non-diabetic nephropathy.

BACKGROUND: The objective of this study was to examine the effects of angiotensin II receptor blocker losartan versus the calcium channel blocker amlodipine on proteinuria, renal haemodynamics, glomerular sieving and tubular function in hypertensive patients with non-diabetic nephropathy. METHODS: The study design was a prospective, double blind, placebo controlled, randomized crossover trial with amlodipine and losartan. Renal parameters were measured at baseline and at the end of each 4-week active treatment period. Fifteen patients with a diagnosis of non-diabetic renal disease and hypertension were included. RESULTS: Mean arterial blood pressure decreased from 123+/-13 mmHg at baseline to 113+/-10 mmHg (P<0.01) on losartan and to 114+/-10 mmHg on amlodipine (P<0.01). Urinary albumin excretion significantly decreased from 3510+/-2586 mg/24 h at baseline to 2684+/-2051 mg/24 h (P<0.01) on losartan and increased non-significantly to 3748+/-3355 mg/24 h on amlodipine. Filtration fraction significantly decreased from a baseline value of 22.8+/-9.3% to 21.2+/-10.2% (P<0.05) on losartan and increased to 23.6+/-8.9% (ns) on amlodipine. Either drug did not significantly alter glomerular sieving of neutral dextrans. CONCLUSION: Our results demonstrate that losartan, but not amlodipine, decreased albumin excretion in hypertensive patients with non-diabetic nephropathy.

Adult↗
PubMed1998

Insulin resistance is a common denominator of post-transplant diabetes mellitus and impaired glucose tolerance in renal transplant recipients.

BACKGROUND: Post-transplant diabetes mellitus is a known complication of steroid therapy in renal transplant recipients. Both insulin resistance and insulin deficiency have been shown to be necessary for development of post-transplant diabetes mellitus. It is not known whether recipients with impaired glucose tolerance have similar degree of insulin resistance or deficient insulin response as recipients with post-transplant diabetes mellitus. METHOD: To address this question, we used an oral glucose tolerance test to categorize 46 renal transplant recipients on triple immunosuppressive medication to groups with normal glucose tolerance, impaired glucose tolerance or post-transplant diabetes mellitus. Insulin sensitivity was measured using a hyperinsulinaemic euglycaemic clamp. Insulin response was calculated from the increase in serum insulin concentration during the oral glucose tolerance test. RESULTS: Twenty-five were categorized to normal glucose tolerance, 15 to impaired glucose tolerance and six to post-transplant diabetes mellitus. There were no statistically significant differences between the groups regarding prednisolone dose, azathioprine dose, use of beta-blocker, age, gender, weight, waist-hip ratio, body mass index, donor source, smoking habits, or first-degree relatives with histories of diabetes mellitus. The impaired glucose tolerance and post-transplant diabetes mellitus groups showed a significant reduction in insulin-stimulated glucose disposal rate (mg/kg.min) compared to the normal glucose tolerance group (4.6 +/- 1.6 and 3.4 +/- 1.3 respectively vs 7.1 +/- 2.4, P < 0.05). The insulin response (picomol/l) was not different between the normal glucose tolerance and impaired glucose tolerance groups but was significantly reduced in the post-transplant diabetes mellitus group (448 +/- 310 and 450 +/- 291 respectively vs 170 +/- 128, P < 0.05). CONCLUSION: Insulin resistance is a common denominator of post-transplant diabetes mellitus and impaired glucose tolerance in renal transplant recipients.

Adult↗
PubMed1997

Gas exchange and neurohumoral response to exercise: influence of the exercise protocol.

Maximal oxygen uptake varies with the exercise protocol, but the extent to which hormonal and metabolic responses to exercise are influenced by the exercise protocol has not been precisely defined. Twelve healthy subjects underwent maximal exercise testing using two incremental bicycle tests with individualized, identical work rate increments between 40 and 70 W. One protocol employed a 1-min and the other a 3-min duration per stage. Expiratory gas and venous blood were sampled at regular intervals for metabolic and hormonal analysis. Exercise duration for the 1-min and 3-min protocols was 6.0 +/- 0.1 and 14.3 +/- 0.3 min, respectively (P < 0.001). Significantly higher values were observed for peak VO2 and maximal ventilation during the 3-min protocol compared with the 1-min protocol (41.1 +/- 1.8 vs 38.3 +/- 1.6 ml.kg-1.min-1, P < 0.001; and 104.9 +/- 8.0 vs 97.2 + 5.7 l.min-1, P < 0.05, for peak VO2 and peak ventilation, respectively). However, the maximal workload achieved was higher during the 1-min versus the 3-min protocol (330 + 24 vs 280 + 21 W, P < 0.01). No differences were observed for maximal heart rate or blood pressure, whereas maximal plasma lactate was roughly twice as high during the 3-min compared with the 1-min protocol (7.5 +/- 0.8 vs 3.8 +/- 0.5 mmol.l-1, P < 0.001). Norepinephrine, epinephrine, dopamine, and growth hormone levels were generally higher throughout exercise during the 3-min compared with the 1-min protocol. When expressed as a percentage of peak VO2, however, differences in catecholamine levels were not observed. Endothelin levels did not change. We conclude that the exercise protocol profoundly influences exercise capacity as well as the metabolic and hormonal response to exercise and should be considered when using these variables to evaluate an intervention.

Adult↗
PubMed1997

Exposure to hydrogen fluoride: an experimental study in humans of concentrations of fluoride in plasma, symptoms, and lung function.

OBJECTIVES: To study the absorption of inhaled hydrogen fluoride (HF) by measuring plasma fluorides and HF concentrations in the breathing zone during exposure to HF. A possible dose-effect relation was investigated by following airway symptoms and lung function-that is, forced expiratory volume in one second (FEV1) and forced vital capacity (FVC)-during and after exposure to HF. METHODS: 20 healthy, male volunteers were exposed for one hour to constant HF concentrations that ranged from 0.2 to 5.2 mg/m3; these concentrations are known to occur among potroom workers in the primary aluminium industry. Plasma fluorides were analysed before, during, and after exposure. Symptoms from the eyes and the upper and lower airways were registered and graded from 1 to 5 with a standardised questionnaire. RESULTS: The total symptom score was significantly increased at the end of exposure for all the subjects as a group (P < 0.01) and for the group exposed to HF below the present Norwegian standard for total fluorides 0.6 mg/m3 (P = 0.05). No change was detected in FEV15 although a significant decrease was found in FVC in the group exposed to fluorides below the hygienic standard (n = 9) and for the entire group (n = 23). Almost all the symptoms had disappeared four hours after the end of exposure. Symptom scores from the upper airways were significantly correlated with the HF concentration (r = 0.62, P = 0.002), the change in plasma fluoride concentration (delta C) (r = 0.51, P = 0.01), and the maximum plasma fluoride concentration (Cmax) (r = 0.42, P = 0.05). A significant correlation was also found between the total symptom score for airways and the HF concentration. CONCLUSIONS: The present study showed a strong relation between inhaled HF and concentrations of fluoride in plasma. Upper airway and eye symptoms occurred after one hour of exposure to HF even when below the Norwegian hygienic standard for fluorides.

Administration, Inhalation↗
PubMed1996

Pancreatic development and maturation of the islet B cell. Studies of pluripotent islet cultures.

Pancreas organogenesis is a highly regulated process, in which two anlage evaginate from the primitive gut. They later fuse, and, under the influence of the surrounding mesenchyme, the mature organ develops, being mainly composed of ductal, exocrine and endocrine compartments. Early buds are characterized by a branching morphogenesis of the ductal epithelium from which endocrine and exocrine precursor cells bud to eventually form the two other compartments. The three compartments are thought to be of common endodermal origin; in contrast to earlier hypotheses, which suggested that the endocrine compartment was of neuroectodermal origin. It is thus generally believed that the pancreatic endocrine-lineage possesses the ability to mature along a differentiation pathway that shares many characteristics with those of neuronal differentiation. During recent years, studies of insulin-gene regulation and, in particular, the tissue-specific transcriptional control of insulin-gene activity have provided information on pancreas development in general. The present review summarizes these findings, with a special focus on our own studies on pluripotent endocrine cultures of rat pancreas.

Amyloid↗
PubMed1996

Breast cancer incidence subsequent to surgical reduction of the female breast.

The incidence of breast cancer among 1240 women who were treated surgically for breast hypertrophy in Copenhagen, Denmark between 1943 and 1971 was determined and compared with age- and calendar period-specific rates for the Danish female population. A total of 32 cases of breast cancer had developed by the end of 1990; the expected number was 52.55, yielding a relative risk (RR) of 0.61 [95% confidence interval (CI) 0.42-0.86]. The greatest reduction in risk was observed for women who had 600 g or more of breast tissue removed (RR=0.30; 95% CI 0.10-0.69). This suggests that the number of potential foci is important for cancer development in the female breast. In the group of women who were operated on before the age of 20, four cases of breast cancer developed, compared with 2.23 expected cases, to give an RR of 1.79, suggesting that the aetiology of their breast hypertrophy may be different from that for the rest of the group.

Adult↗
PubMed1996

Delivery of newly synthesized tubulin to rapidly growing distal axons of sympathetic neurons in compartmented cultures.

Growing axons receive a substantial supply of tubulin and other proteins delivered from sites of synthesis in the cell body by slow axonal transport. To investigate the mechanism of tubulin transport most previous studies have used in vitro models in which the transport of microtubules can be visualized during brief periods of growth. To investigate total tubulin transport in neurons displaying substantial growth over longer periods, we used rat sympathetic neurons in compartmented cultures. Tubulin synthesized during pulses of [35S]methionine was separated from other proteins by immunoprecipitation with monoclonal antibodies to alpha and beta tubulin, further separated on SDS-PAGE, and quantified by phosphorimaging. Results showed that 90% of newly synthesized tubulin moved into the distal axons within 2 d. Furthermore, the leading edge of tubulin was transported at a velocity faster than 4 mm/d, more than four times the rate of axon elongation. This velocity did not diminish with distance from the cell body, suggesting that the transport system is capable of distributing newly synthesized tubulin to growth cones throughout the axonal tree. Neither diffusion nor the an mass transport of axonal microtubules can account for the velocity and magnitude of tubulin transport that was observed. Thus, it is likely that most of the newly synthesized tubulin was supplied to the growing axonal tree in subunit form such as a heterodimer or an oligomer considerably smaller than a microtubule.

Animals↗
PubMed1996

The use of isolated lung cells in in vitro pulmonary toxicology: studies of DNA damage, apoptosis and alteration of gene expression.

Isolated lung cells constitute a valuable system for studying mechanisms involved in chemically induced toxicity in the lung. Different lung cells isolated from various species may be studied. Bronchiolar Clara and alveolar type 2 cells produce important lung-specific proteins, hold a major role in the metabolism of xenobiotics and serve as progenitor cells for other lung cell types. They are possible target cells in lung carcinogenesis. Alveolar macrophages play an important role in lung defence and in inflammatory responses. In the present study we have characterised chemically induced DNA damage, apoptosis, changes in cell cycle progression, transformation and alterations in gene expression in these specific lung cells isolated from rat, rabbit and human. Major differences between the cell types and the various species in the induction of DNA damage by chemicals were found, as measured by the 32P-postlabelling and alkaline filter elution techniques. Benzo(a)pyrene and hydrogen fluoride were found to induce apoptosis in the isolated cells as measured by microscopical analysis and flow cytometry. The function of various important tissue- or cell type specific proteins (CYP 2B1, Clara cell protein) and/or cellular signal transduction pathways constitute important targets that may be affected by exposure to toxic compounds. Using immunological and molecular techniques the differential expression of specific proteins/RNAs and their activity can be studied. Among other proteins, c/ebp is involved in the regulation of transcription at the end of signal pathways. The protein is differentially expressed in rat lung cells and thus could be suitable for studying differential toxic effects in various lung cells. In humans, bronchoalveolar lavage (BAL) fluid from human volunteers can be readily obtained and examined after exposure to different chemical compounds. An increase in the percentage of CD3-positive cells (T-lymphocytes) was found after exposure to hydrogen fluoride. The number of certain cell types and cytokines may be used to estimate the degree of inflammatory reaction. In conclusion, the use of in vitro data including the use of specific, primary human lung cell types may contribute considerably to the quality of risk assessment, together with in vivo data from animals and man.

Animals↗
PubMed1995

Proteolytic processing mechanisms in the biosynthesis of neuroendocrine peptides: the subtilisin-like proprotein convertases.

The recent discovery of a novel family of precursor processing endoproteases has greatly accelerated progress in understanding the complex mechanisms underlying the maturation of prohormones, neuropeptides, and many other precursor-derived proteins. At least six members of this family have been found thus far in mammalian species, several having alternatively spliced isoforms, and related enzymes have been identified in many invertebrates, including molluscs, insects, nematodes, and coelenterates. The proprotein convertases are all dependent on calcium for activity and all possess highly conserved subtilisin-like domains with the characteristic catalytic triad of this serine protease (ordered Asp, His, and Ser along the polypeptide chain). Two members of this family, PC2 (SPC2) and PC1/PC3 (SPC3), appear to play a preeminent role in neuroendocrine precursor processing. Both convertases are expressed only in the brain and in the extended neuroendocrine system, while another important family member--furin/PACE (SPC1)--is expressed more ubiquitously, in almost all tissues, and at high levels in liver. SPC2 and SPC3 exhibit acidic pH optima and other properties which enhance their activity in the acidic, calcium-enriched environment of the dense-core secretory granules of the regulated pathway in neuroendocrine cells, while furin has a neutral pH optimum and is localized predominantly to the trans Golgi network where it is retained by a C-terminal transmembrane domain. Furin processes a wide variety of precursors in the constitutive pathway, such as those of growth factors, receptors, coagulation factors, and viral glycoproteins. Recent findings on the processing of proopiomelanocortin, proinsulin, proglucagon, and several other neuroendocrine precursors by SPC2 and SPC3 are discussed, along with information on the structure, properties, evolution, developmental expression, and regulation of the convertases. An inherited defect in the fat/fat mouse which affects the processing of proinsulin, and probably also many other prohormones, due to a point mutation in carboxypeptidase E has recently been identified and has begun to provide new insights into the functional integration of the individual processing steps.

Animals↗