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Biomedical subjects

L Carlsson

Publications and source records attributed to L Carlsson.

At least 55 records · Page 3Linked to original sources

Growth-inhibitory effect of prostasomes on prostatic cancer cell lines in culture.

OBJECTIVES: Prostasomes, small corpuscular organelles derived from the prostate gland, were isolated from human seminal plasma for incubation with three human prostatic cancer cell lines: DU145, PC3, and LNCaP. The aim of this study was to establish any possible growth-inhibitory effect of prostasomes on prostatic cancer cell lines. METHODS: Prostasomes were isolated from human seminal plasma. We determined their growth-inhibitory effect on the prostatic cancer cell lines by a fluorimetric cytotoxic assay technique. The results obtained were presented as survival index, defined as a percentage of control cultures, after compensation for blank values. RESULTS: The prostasomes exhibited a dose-dependent growth-inhibitory effect on the cells, cell line DU145 being the most sensitive one displaying a survival index (mean +/- SD) of 46+/-10% at 200 microg prostasomes (protein)/ml while the other two cell lines displayed survival indices of 93+/-6 and 93+/-9%. Heat treatment of the prostasomes abolished their growth-inhibitory effect on the cell lines, whereas 24-hour dialysis of the prostasomes did not affect their activity in this regard. Seminal plasma devoid of prostasomes exhibited less effect on the growth of these cancer cell lines, and heat treatment of the plasma did not influence the result. However, dialysis of the seminal plasma for 24 h abrogated its growth-inhibitory effect. CONCLUSION: The prostasomal growth-inhibitory effect on the cancer cell lines was different from that of the prostasome-free seminal plasma. The former effect was most probably of a proteinaceous nature while the latter effect was most likely due to the divalent cation composition of seminal plasma. In this regard, the zinc ion content seemed to be decisive.

Cell Culture Techniques↗

[Use of new materials results in improved prostheses. Metals, polymers, ceramics and composite materials extend durability].

Advances in our knowledge of multifactorial host-biomaterial interactions have improved the outcome of joint replacement surgery. Fixation and aseptic loosening are the principal foci of research. Wear debris from articulating parts is a major cause of prosthetic loosening. Tri-biological analysis, and the development of new materials and surface finishes have reduced wear particle production. The use of modular implants increases flexibility and facilitates surgical fit, but also introduces new problems such as dissociation of components, corrosion and wear. In the future, as design, biomaterials and surgical technique become further optimized, arthroplasty may also become an alternative for use in younger patients.

Arthroplasty, Replacement↗

Nestin is expressed during development and in myotendinous and neuromuscular junctions in wild type and desmin knock-out mice.

Desmin, the main component of intermediate filaments (IFs) in mature skeletal muscle, forms an interlinking scaffold around myofibrils with connections to the sarcolemma and the nuclear membrane. Desmin is enriched in neuromuscular and myotendinous junctions. Mice lacking the desmin gene develop normally and reproduce. However, postnatally they develop a cardiomyopathy and a dystrophy in highly used muscles. We have investigated whether and how neuromuscular and myotendinous junctions are affected and whether nestin compensates for the lack of desmin in the knock-out (K/O) mice. We show that neither neuromuscular nor myotendinous junctions were markedly affected in the desmin K/O mice. In neuromuscular junctions nestin was present between the postjunctional folds and the subneural nuclei and between the nucleus and the myofibrillar cytoskeleton. In myotendinous junctions nestin was present between myofibrils at the Z-disc level and in longitudinal strands close to and at the junction. Nestin expression at these specialized sites, as well as during myogenesis and myofibrillogenesis, is independent of the presence of desmin. In desmin K/O mice nestin was also found in regenerating myofibers. The presence of nestin at neuromuscular and myotendinous junctions might provide enough strength for preservation and organization of the junctional areas, although desmin is lacking.

Aging↗

[New materials improve joint prostheses. Metals, polymers, ceramics and composite materials extend the durability].

Advances in our knowledge of multfactorial host-biomaterial interactions have improved the outcome of joint replacement surgery. Fixation and aseptic loosening are the principal foci of research. Wear debris from articulating parts is a major cause of prosthetic loosening. Tribological analysis, and the development of new materials and surface finishes have reduced wear particle production. The use of modular implants increases flexibility and facilitates surgical fit, but also introduces new problems such as dissociation of components, corrosion and wear. In the future, as design, biomaterials and surgical technique become further optimised, arthroplasty may also become an alternative for use in younger patients.

Animals↗

Identification of dipeptidyl peptidase IV as the antigen of a monoclonal anti-prostasome antibody.

BACKGROUND: Our purpose was to elucidate the identity of an antigen of a monoclonal antibody against prostasomes, which are prostate-derived organelles occurring freely in human semen. METHODS: Monoclonal antibodies against prostasomes were generated by intrasplenic immunization to allow better characterization of the prostasomes. One third of the resulting antibodies bound to a 110-kDa prostasomal protein on Western blots. RESULTS: The antigen was purified from seminal prostasomes by anion exchange chromatography and gel electrophoresis. After in-gel digestion and amino-acid sequence analysis of selected peptides, it was identified as dipeptidyl peptidase IV (DPP IV), EC 3.4.14.5, also known as T-cell-activating antigen CD26. No DPP IV could be detected in the supernatant obtained after preparative ultracentrifugation of the prostasomes. CONCLUSIONS: DPP IV/CD26 is structurally bound to prostasomes rather than being free in seminal fluid. This enzyme may play a critical role in the promoting effect of prostasomes on forward motility of spermatozoa.

Amino Acid Sequence↗

Some factors affecting enantiomeric impurity determination by capillary electrophoresis using ultraviolet and laser-induced fluorescence detection.

The key factors influencing enantiomer trace determination were investigated; these include resolution capillary diameter, limit of detection, linear range and type of detection. Chiral reagents, (+)- and (-)-1-(9-fluorenyl)ethyl chloroformate (FLEC), were employed as probes to demonstrate the influence of the variables. In order to find the best resolution, separation variables were optimized in both capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MEKC) modes by the application of factorial design experiments. A highly efficient chiral separation of the (+/-)-FLEC, derivatized with nonchiral amino acids, was achieved when using gamma-cyclodextrin as the chiral selector. The benefits of using a small diameter capillary for direct determination of both (+) and (-)-FLEC impurity (0.05-0.1% area/area) were demonstrated using UV detection and applying a sample stacking condition. A frequency-doubled argon ion laser (244 nm) was used as light source for laser-induced fluorescence (LIF) detection. Excitation light was provided by means of an optical fiber directed into the Hewlett Packard 3D capillary cartridge. The signals from UV and LIF were monitored simultaneously. The application of LIF detection greatly improved sensitivity and linear range. Further, as a consequence of the increased sensitivity, sample loading could be decreased, which led to an improvement of separation efficiency. Direct determination of 0.005% impurity could be achieved within the linear range.

Chromatography, Micellar Electrokinetic Capillary↗

Birth weight, adulthood BMI, and subsequent weight gain in relation to leptin levels in Swedish women.

OBJECTIVE: Leptin seems to be involved in the regulation of energy balance, although little is known about the epidemiology of leptin with respect to prediction of weight gain and incidence of obesity-related diseases. The dual aim of this study is to document characteristics of leptin after long-term storage, and to describe its relation to body weight, from birth to old age, in an ongoing prospective study. RESEARCH METHODS AND PROCEDURES: A population-based sample of Swedish women was first examined at the ages of 38 to 60 and re-examined 24 years later. This study used 1358 frozen serum samples that had been stored 29 years for analysis of leptin concentrations and their relation to body weight history. RESULTS: Leptin values obtained from stored samples showed the same correlation with relative weight as that seen in a contemporary sample with similar demographic characteristics. Lower self-reported birth weight was associated with higher leptin levels in adulthood (p = 0.01), controlling for age and adult BMI. Prospective analyses revealed that high leptin in 38 to 46-year-olds predicted subsequent long-term weight gain (p = 0.003), although no significant associations were seen in women initially aged 50 or older. DISCUSSION: It is feasible to use frozen serum for studying leptin in relation to obesity and related developments many years later. High leptin level was a risk factor for subsequent weight gain in 38- and 46-year-old women. Retrospective analyses involving birth weight suggest that leptin resistance in adulthood might have fetal origins.

Adult↗

Metabolic fates in humans of linamarin in cassava flour ingested as stiff porridge.

Insufficiently processed products from cassava roots may contain residual amounts of cyanogenic glucosides, mainly linamarin. The fate of orally ingested linamarin was studied following a meal of cassava porridge prepared from cassava flour from southern Tanzania with 82 mg cyanide equivalents (3035 micromol) of linamarin per kg dry weight. Following ingestion of amounts of porridge containing 243-571 micromol linamarin by 15 healthy adults a mean (range) of 21% (1-47%) of the linamarin ingested was excreted in the urine within 24 hours and a mean of 1% in the next 24 hours. Serum thiocyanate, the main cyanide metabolite, increased in all subjects from a mean (+/-SD) of 34+/-26 to 78+/-28 micromol/litre (P < 0.001). In a second group of seven subjects we found that the ingestion of porridge with a mean (range) of 431 micromol (203-669%) of linamarin resulted in a mean linamarin excretion of 127 micromol/litre and an excess thiocyanate excretion of 118 micromol/litre and that 216 micromol was unaccounted for. We conclude that less than one-half of orally ingested linamarin is converted to cyanide and hence thiocyanate, about one-quarter is excreted unchanged and another quarter is metabolized into an as yet unknown compound.

Adult↗

Hydroxyapatite coating versus cemented fixation of the tibial component in total knee arthroplasty: prospective randomized comparison of hydroxyapatite-coated and cemented tibial components with 5-year follow-up using radiostereometry.

Fifty-three consecutive patients (57 knees; mean age, 69 years) entered a prospective randomized study to compare the fixation of hydroxyapatite (HA)-coated (29 knees) with cemented (28 knees) tibial components in the Tricon II total knee arthroplasty. The quality of the fixation during 5 years postoperatively was evaluated with radiostereometric analysis (RSA). Three HA-coated implants were revised: 2 owing to infection, and 1 owing to early delamination of the coating and clinical loosening. Eight patients (9 knees) died, 1 patient sustained a stroke, and 1 patient refused investigations after 1 year. In the 40 patients (19 HA-coated, 21 cemented) remaining at 5 years, the magnitude of the micromotion between the HA-coated and cemented groups did not differ. The HA-coated implants displayed most of the migration within the initial 3 months then stabilized, whereas the cemented implants showed an initially lower, but over time continuously increasing migration. Between 1 and 2 years, 4 of 24 HA-coated and 10 of 23 cemented implants migrated >0.2 mm and were categorized unstable, which has been shown to have a prognostic value as regards future aseptic loosening. Progressive radiolucent lines developed in 2 cemented knees, which both were categorized unstable. If HA-coated implants can sustain the forces that threaten the fixation in the early period after implantation, a strong and enduring fixation may be obtained.

Adult↗

Eosinophilic inflammation assessed by induced sputum in corticosteroid-dependent asthma.

Sputum induction can be used to study airway inflammation in asthmatics. However, it has not been used in patients with corticosteroid-dependent asthma requiring long-term oral corticosteroids. The aim of the study was to assess the number of eosinophils and the levels of eosinophil cationic protein (ECP) in sputum of 17 corticosteroid-dependent asthmatics by comparison with nine mild untreated asthmatics, 10 moderate asthmatics receiving inhaled steroids (ICS) and 11 healthy subjects. In the 17 corticosteroid-dependent asthmatics, we examined sputum eosinophil markers on two occasions and correlated with the control of asthma. Eosinophils were undetectable in controls and were detected in 63.8% of asthmatics. There were no significant differences between the three groups of asthmatics. ECP levels were significantly increased in ICS or corticosteroid-dependent asthmatics by comparison to controls and mild asthmatics. There was no significant difference between ICS and corticosteroid-dependent asthmatics. During follow-up, corticosteroid-dependent asthmatics with a controlled disease had no significant change in eosinophil numbers or ECP levels. On the other hand, corticosteroid-dependent asthmatics with recent exacerbations had a non-significant increase in eosinophil numbers and a significant increase in ECP levels. This study shows that ECP levels may be more accurate than eosinophil numbers in assessing exacerbations in corticosteroid-dependent asthmatics.

Adolescent↗

Greater increase in cytokine concentration after salvage with filtered whole blood than with washed red cells, but no difference in postoperative hemoglobin recovery.

BACKGROUND: Inflammatory mediators are released in association with intraoperative and postoperative salvage of blood. Whether these mediators (cytokines) participate in the modulation of erythropoiesis or not has been investigated. STUDY DESIGN AND METHODS: Twenty-seven patients who were to undergo total knee replacement surgery were randomly assigned to postoperative blood salvage with either filtered whole blood or washed red cells. Patients with postoperative blood loss <400 mL were considered a control group. The control group did not receive any transfusions. Plasma concentrations of the anaphylatoxin C3a, the C5b-9 terminal complement complex, and the cytokines interleukins 6 and 8, hemoglobin, reticulocytes, and red cell volume fraction in the patients were repeatedly analyzed before and after surgery. RESULTS: Significantly increased concentrations of interleukin 6 appeared in all three groups, which was interpreted as a response to the surgical trauma. The increase was significantly greater in the group that received filtered whole blood after return of shed blood. The recovery of hemoglobin levels did not differ in the groups. CONCLUSION: The transfusion of filtered whole blood leads to the formation of interleukin 6 in the circulation, but postoperative hemoglobin recovery was similar in all groups.

Aged↗

Subsidence of a non-polished stem in revisions of the hip using impaction allograft. Evaluation with radiostereometry and dual-energy X-ray absorptiometry.

We revised 24 consecutive hips with loosening of the femoral stem using impaction allograft and a cemented stem with an unpolished proximal surface. Repeated radiostereometric examinations for up to two years showed a slow rate of subsidence with a mean of 0.32 mm (-2.0 to +0.31). Fifteen cases followed for a further year showed the same mean subsidence after three years, indicating stabilisation. A tendency to retroversion of the stems was noted between the operation and the last follow-up. Retroversion was also recorded when displacement of the stem was studied in ten of the patients after two years. Repeated determination of bone mineral density showed an initial loss after six months, followed by recovery to the postoperative level at two years. Defects in the cement mantle and malalignment of the stem were often noted on postoperative radiographs, but did not correlate with the degrees of migration or displacement. After one year, increasing frequency of trabecular remodelling or resorption of the graft was observed in the greater trochanter and distal to the tip of the stem. Cortical repair was noted distally and medially (Gruen regions 3, 5 and 6). Migration of the stems was the lowest reported to date, which we attribute to the improved grafting technique and to the hardness of the graft.

Absorptiometry, Photon↗

Expression of prostasome-like granules by the prostate cancer cell lines PC3, Du145 and LnCaP grown in monolayer.

Prostasomes are a granular type of secretory product in the human prostate gland cells. It is not known, whether in vitro grown cells derived from human prostate cancers also express prostate secretory components containing granules with properties similar to the prostasomes. Therefore, we carried out the present investigation and found that cytospins of in vitro grown PC3, DU145 and LNCaP cells generally expressed a granular secretion. DU145 demonstrated the highest ratio of cells with granules (about 90%), while cytospins of PC3 and LNCaP contained less stained cells (50-70%). Purified granules from PC3 cells were immunoreactive with a monoclonal antibody (mAb78) originally raised against human seminal prostasomes. The PC3 granules also shared the property with human seminal prostasomes having an elevated UV260/UV280 absorbance ratio. On the other hand we found a low aminopeptidase activity in PC3 granules contrary to that of human prostasomes. Prostasomes may form a heterogeneous group with different properties due to the source from which they are isolated and perhaps it is justified to recognize them as different members of a prostasome family.

Aminopeptidases↗

Expression of the LIM-homeobox gene LH2 generates immortalized steel factor-dependent multipotent hematopoietic precursors.

The genes controlling self-renewal and differentiation in the hematopoietic system are largely unknown. The LIM-homeobox genes are known to be important for asymmetric cell divisions and differentiation of specific cell types and organs. One member of this family, LH2, is expressed in fetal liver at the time of active hematopoiesis. Therefore, we have assessed the function of LH2 during the formation and initial expansion of the hematopoietic system by differentiating LH2-transduced embryonic stem (ES) cells in vitro. This procedure generated multipotent hematopoietic precursor cell (HPC) lines that required Steel factor for growth. HPC lines have been maintained in an undifferentiated state in culture for >7 months. Other growth factors tested efficiently induce terminal differentiation of HPCs into various mature myeloid lineages. Steel factor is also required and acts synergistically with the other growth factors to generate multilineage colonies from the HPCs. These HPC lines express transcription factors that are consistent with an immature progenitor, and the pattern of cell surface marker expression is similar to that of early fetal multipotent hematopoietic progenitors. Collectively, these data suggest that the HPC lines represent an early fetal multipotent hematopoietic progenitor, and suggest a role for LH2 in the control of cell fate decision and/or proliferation in the hematopoietic system.

Animals↗

Biomechanical characterization of osseointegration: an experimental in vivo investigation in the beagle dog.

This study reports the results of torsion tests, pull-out tests, and lateral loading tests on osseointegrated commercially pure titanium fixtures. The tests were performed in vivo on six beagle dogs. Three fixtures, each with a diameter of 3.7 mm, were installed bilaterally in the tibia of each animal. The mean maximal pull-out load was 1.55 kN (n = 4), the mean maximal lateral transverse load was 0.21 kN (n = 2), the mean maximal lateral axial load was 0.18 kN (n = 2), the mean breakpoint torque was 0.31 Nm (n = 3), and the mean maximal torque was 0.43 Nm (n = 3). The torsion test revealed an almost immediate plastic deformation of the interface between the implant and bone; this indicates that although the contact between the bone and the implant is close, there is no strong bond, at least not in shear. The major transfer of load from the implant to the surrounding bone tissue must therefore depend on the design of the implant. A histological evaluation with measurements of the amount of bone in contact with the fixtures was performed. By the use of the histological and mechanical data, it is possible to estimate shear stresses in bone tissue (pull-out test) and in the interface (torque test). The mean maximal shear stress in bone tissue in the pull-out tests was 100 MPa (n = 4); the mean shear stress in the interface was 4.3 MPa (n = 3) in the torsion tests at the breakpoint torque and was 6.0 MPa (n = 3) at the maximal torque. It was also possible to estimate the shear modulus of elasticity in the pull-out and torque tests. The mean shear modulus in pull-out was 119 MPa (n = 4), and the mean apparent shear modulus in torsion was 9 kPa (n = 3) for an assumed interface thickness of 100 nm and was 86 kPa (n = 3) for an assumed interface thickness of 1,000 nm.

Animals↗

Ceramic coating improves tibial component fixation in total knee arthroplasty.

Forty uncemented total knee arthroplasties (36 patients) were randomly allocated to a Miller-Galante II prosthesis with or without hydroxyapatite/tricalcium phosphate (HA/TCP) coating on the titanium fiber mesh on the undersurface of the tibial component. The patients were followed for 2 years with repeated radiostereometric examinations. After 2 years, the HA/TCP tibial components displayed smaller anterior-posterior tilt and less subsidence. The mean value of maximal total point motion was small: 0.5 mm in both groups. At 24 months, there were more radiolucent lines under the tibial tray and around the stem in the uncoated group, but the clinical results did not differ. HA/TCP coating on the undersurface of the tibial component improved the stability and seemed to improve the quality of the interface between the tibial component and the bone.

Aged↗

Identification of a fetal hematopoietic precursor with B cell, T cell, and macrophage potential.

Despite years of investigation, precursor-progeny relationships within the developing lymphoid lineages of the hematopoietic system remain poorly defined. We have characterized the potential of precursors found within a subpopulation of fetal liver defined by AA4.1 and Fc gammaRII/III expression and predominantly restricted to lymphoid and macrophage development. When cultured in methylcellulose with appropriate cytokines, AA4.1+/Fc gammaR+ precursors generate colonies consisting of various lineages, including the combination of B cell, T cell, and macrophage. Retroviral marking studies showed that the lymphoid cells and macrophages within these colonies arise from a common precursor. These results demonstrate the presence of a common precursor with B cell-, T cell-, and macrophage-restricted potential and as such define an early restriction point within the fetal lymphoid system.

Animals↗

Hormonal regulation of liver fatty acid-binding protein in vivo and in vitro: effects of growth hormone and insulin.

Liver fatty acid-binding protein (LFABP) is an abundant protein in hepatocytes that binds most of the long chain fatty acids present in the cytosol. It is suggested to be of importance for fatty acid uptake and utilization in the hepatocyte. In the present study, the effects of bovine GH (bGH) and other hormones on the expression of LFABP and its messenger RNA (mRNA) were studied in hypophysectomized rats and in vitro using primary cultures of rat hepatocytes. One injection of bGH increased LFABP mRNA levels about 5-fold after 6 h, but there was no effect of this treatment on LFABP levels. However, 7 days of bGH treatment increased both LFABP mRNA and LFABP protein levels 2- to 5-fold. Female rats had higher levels of LFABP than male rats. Hypophysectomy of female rats, but not that of male rats, decreased LFABP levels markedly. Treatment of hypophysectomized rats with bGH for 7 days as two daily injections or as a continuous infusion increased LFABP levels to a similar degree. This finding indicates that the sex difference in the expression of LFABP is not regulated by the sexually dimorphic secretory pattern of GH. Neither insulin nor insulin-like growth factor I treatment of hypophysectomized rats for 6-7 days had any effect on LFABP mRNA or LFABP levels. In vitro, bGH dose-dependently increased the expression of LFABP mRNA, but only in the presence of insulin. Insulin alone had a marked dose-dependent effect on LFABP mRNA levels and was of importance for maintaining the expression of LFABP mRNA during the culture. Incubation with bGH increased LFABP mRNA levels within 3 h. GH had no effect on LFABP mRNA levels in the presence of actinomycin D, indicating a transcriptional effect of GH. Incubation with glucagon in vitro decreased LFABP mRNA levels markedly, indicating that glucagon, in contrast to GH, has an effect opposite that of insulin on LFABP mRNA expression. It is concluded that GH is an important regulator of LFABP in vivo and in vitro. In contrast to the effect of GH on insulin-like growth factor I mRNA, the presence of insulin was a prerequisite for the effect of GH on LFABP mRNA expression in vitro. The results emphasize the role of GH in the regulation of hepatic fatty acid metabolism.

Animals↗