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Biomedical subjects

C B Andersen

Publications and source records attributed to C B Andersen.

At least 37 records · Page 2Linked to original sources

Apoptosis in acute pulmonary allograft rejection and cytomegalovirus infection.

Apoptosis is a form of programmed cell death, characterized by activation of endonucleases that cleave DNA into oligonucleosomal fragments, which can be identified by in situ terminal deoxyribonucleotide transferase-mediated dUTP nick-end labeling (TUNEL). This process has recently been implicated in cardiac and hepatic allograft rejection, and we investigated its contribution to acute pulmonary allograft rejection and cytomegalovirus (CMV) pneumonitis by in situ TUNEL of transbronchial biopsy specimens. In situ TUNEL was performed on 70 transbronchial biopsy samples collected from 25 pulmonary allograft recipients for diagnosis of acute rejection or CMV pneumonitis, and the number of apoptotic nuclei/mm2 was correlated with the rejection grade (International Society of Heart and Lung Transplantation classification). During acute pulmonary allograft rejection, apoptotic nuclei were demonstrated in pulmonary parenchymal cells and mononuclear infiltrating cells, and the number of apoptotic cells was positively correlated with the rejection grade. In addition, a marked increase in the density of apoptotic cells was found in pulmonary allografts with CMV pneumonitis. We conclude that apoptosis contributes to cell death during acute pulmonary allograft rejection and CMV infection.

Acute Disease↗

Validity of histopathological grading of articular cartilage from osteoarthritic knee joints.

OBJECTIVES: To determine the validity of the histological-histochemical grading system (HHGS) for osteoarthritic (OA) articular cartilage. METHODS: Human articular cartilage was obtained from macroscopically normal (n = 13) and OA (n = 21) knee joints. Sections of central and peripheral regions of normal samples were produced. Sections of regions containing severe, moderate, and mild OA changes were produced from each OA sample. A total of 89 sections were graded by means of the HHGS (0-14) twice by three observers. RESULTS: Average scores for regions designated severe (8.64) and moderate (5.83) OA were less than the expected (10-14 and 6-9, respectively) according to the HHGS, whereas average scores for the region designated mild (5.29) OA and central and peripheral regions (2.19) of normal cartilage were higher than expected (2-5 and 0-1, respectively). The HHGS was capable of differentiating between articular cartilage from macroscopically normal and OA joints and between the region designated severe OA and other regions. However, the HHGS did not adequately differentiate between regions designated mild and moderate OA. Values for sensitivity, specificity, and efficiency for all regions varied considerably. CONCLUSION: The HHGS is valid for normal and severe OA cartilage, but does not permit distinction between mild and moderate OA changes in articular cartilage.

Adolescent↗

Role of cyclic nucleotide phosphodiesterases in resumption of meiosis.

In the follicles of the mammalian and amphibian ovary, oocyte maturation is arrested at the prophase of the first meiotic division. Prior to ovulation, oocytes reenter the cell cycle, complete the meiotic division, and extrude the first polar body. Work from several laboratories including ours has provided evidence that the cAMP-mediated signal transduction pathway plays an important role in regulation of meiosis, the cyclic nucleotide acting as a negative regulator of maturation. Since cAMP can be regulated both at the level of synthesis and degradation, our laboratory is investigating the role of phosphodiesterases (PDE) in the control of cAMP levels of oocytes. Using pharmacological and molecular tools, we have determined that a PDE3 is the enzyme involved in the control of cAMP levels in the oocytes. In vitro and in vivo studies have established that inhibition of the oocyte PDE3 blocks resumption of a PDE is per se sufficient to cause resumption of meiosis in an amphibian oocyte model. The pathways regulating this PDE isoform expressed in the oocyte is under investigation, as they may uncover the physiological signals controlling meiosis.

3',5'-Cyclic-AMP Phosphodiesterases↗

Protein kinase B/Akt induces resumption of meiosis in Xenopus oocytes.

The activation of protein kinase B/Akt is thought to be a critical step in the phosphoinositide 3-kinase pathway that regulates cell growth and differentiation. Because insulin-like growth factor 1 stimulates the resumption of meiosis in Xenopus laevis oocytes via phosphoinositide 3-kinase activation, we investigated the Akt involvement in this process. Injection of mRNA coding for a constitutively active Akt in Xenopus oocytes induced germinal vesicle breakdown (GVBD) to the same extent as progesterone or insulin treatment. Injection of mRNA coding for the wild type Akt kinase was less effective in stimulating GVBD, whereas Akt bearing a lysine mutation in the catalytic domain that abolishes the kinase activity had no effect. A mutant Akt lacking a membrane-targeting sequence did not induce GVBD, despite high levels of expression and activity. As previously reported for insulin, induction of GVBD by Akt was prevented by incubating the oocytes with cilostamide, an inhibitor specific for the type 3 phosphodiesterase (PDE3), suggesting that the activity of a PDE is required for Akt action. That an increase in PDE activity in the oocyte is sufficient to induce meiotic resumption was demonstrated by expression of an active PDE protein. In addition, the constitutively active Akt caused a 2-fold increase in the activity of the endogenous PDE. These data demonstrate that Akt is in the pathway controlling resumption of meiosis in the Xenopus oocyte and that regulation of the activity of a PDE3 is a step distal to the kinase activation.

3',5'-Cyclic-AMP Phosphodiesterases↗

Definition of MHC and T cell receptor contacts in the HLA-DR4restricted immunodominant epitope in type II collagen and characterization of collagen-induced arthritis in HLA-DR4 and human CD4 transgenic mice.

Rheumatoid arthritis (RA) is an autoimmune disease associated with the HLA-DR4 and DR1 alleles. The target autoantigen(s) in RA is unknown, but type II collagen (CII) is a candidate, and the DR4- and DR1-restricted immunodominant T cell epitope in this protein corresponds to amino acids 261-273 (CII 261-273). We have defined MHC and T cell receptor contacts in CII 261-273 and provide strong evidence that this peptide corresponds to the peptide binding specificity previously found for RA-associated DR molecules. Moreover, we demonstrate that HLA-DR4 and human CD4 transgenic mice homozygous for the I-Abbeta0 mutation are highly susceptible to collagen-induced arthritis and describe the clinical course and histopathological changes in the affected joints.

Amino Acid Sequence↗

Inducible nitric oxide synthase (iNOS) in the human heart: expression and localization in congestive heart failure.

The inducible nitric oxide (NO) synthase (iNOS or NOS2) generates a prolonged release of large amounts of NO which may be cytotoxic and/or inhibit myocyte contractility. It has been suggested that this mechanism specifically contributes to heart failure caused by dilated cardiomyopathy (DCM). To test this hypothesis we compared the myocardial amount and localization of iNOS in myocardial biopsies from patients with heart failure caused by either DCM or ischemic heart disease (IHD). During heart transplantation, myocardial biopsies collected from the diseased heart after explantation were frozen in liquid nitrogen. Twenty-two patients in NYHA class III-IV were included (DCM: n = 8; IHD: n = 14). In each biopsy, iNOS expression was assessed using reverse transcription polymerase chain reaction (RT-PCR), and visualized by immunohistochemistry. iNOS was detected in all biopsies. Intriguingly, the amount of iNOS mRNA (shown as iNOS cDNA normalized to GADPH cDNA) did not differ significantly between the two groups (DCM 30 +/- 7; IHD 20 +/- 6, mean +/- S.E.M., P > 0.05). Similarly, no inter-group differences in the amount of iNOS protein (Western) were observed. iNOS was invariably located to vascular endothelial and smooth muscle cells. In addition, an iNOS reaction in relation to the myocyte membrane was found in 4 of the 22 patients. These four patients (two from each group) had significantly (P < 0.05) higher iNOS/GADPH ratios (54 +/- 20) than patients without myocyte membrane iNOS reaction (17 +/- 15). In conclusion, iNOS is expressed in the myocardium of all patients with heart failure caused by either DCM or IHD. iNOS is located primarily and invariably in the endothelium and vascular smooth muscle cells of the myocardial vasculature and its expression appears to be associated with the condition of heart failure per se rather than related to the heart failure etiology.

Adult↗

Does bronchial artery revascularization influence results concerning bronchiolitis obliterans syndrome and/or obliterative bronchiolitis after lung transplantation?

OBJECTIVE: To study the frequency of histological obliterative bronchiolitis and clinical bronchiolitis obliterans syndrome after en bloc double lung transplantation with bronchial artery revascularization and bilateral lung transplantation without bronchial artery revascularization. METHODS: Primary en bloc double lung transplantation with bronchial artery revascularization using the internal mammary artery as conduit was performed in 62 patients. The frequencies of obliterative bronchiolitis and bronchiolitis obliterans syndrome have been established from transbronchial biopsies and lung function measurements. Results have been analyzed in relation to the arteriographic success of bronchial artery revascularization and have been compared to results from Stanford University, obtained through personal communications. RESULTS: Survival after 1, 2, 3, 4 and 5 years was 85, 81, 69, 69, and 69%, respectively. Fifteen patients developed bronchiolitis obliterans syndrome while seven developed obliterative bronchiolitis. Survival was superior for patients with bronchial artery revascularization classified as complete or incomplete bilateral versus incomplete hemilateral, incomplete poor or failed (P = 0.016, log-rank test). For patients surviving > or = 3 months post-transplant, the post-operative baseline FEV1 was lower for patients who later developed bronchiolitis obliterans syndrome compared to patients who did not (P = 0.007). The development of bronchiolitis obliterans syndrome and obliterative bronchiolitis were both correlated to observation time post-transplant but not to the number of rejections or infections when corrected for observation time. CONCLUSIONS: In a subgroup of lung transplant patients, a process in the transplanted lungs, eventually leading to bronchiolitis obliterans syndrome diagnosis, seems to start in the donor during the transplantation and/or in the early post-operative cause. A comparison with results after bilateral lung transplantation without bronchial artery revascularization suggests that good direct bronchial artery revascularization may postpone the onset of bronchiolitis obliterans syndrome and obliterative bronchiolitis. The positive trend motivates further use of direct bronchial artery revascularization in lung transplantation.

Adolescent↗

Immunohistochemical detection of DNA topoisomerase IIalpha, P-glycoprotein and multidrug resistance protein (MRP) in small-cell and non-small-cell lung cancer.

Non-small-cell lung cancer (NSCLC) and small-cell lung cancer (SCLC) differ significantly in their clinical response to topoisomerase IIalpha (topo-IIalpha)-directed drugs, such as etoposide and teniposide, as NSCLC is virtually insensitive to single-agent therapy, while SCLC responds in two-thirds of cases. Preclinical studies have indicated that resistance to topo-IIalpha drugs depends on topo-IIalpha content and/or activity, the altered-topo-II multidrug resistance phenotype (at-MDR) and/or one of two different drug efflux pumps, P-glycoprotein (P-gp) and the multidrug resistance protein (MRP). Immunohistochemical analysis on paraffin-embedded tissue from 27 cases of untreated NSCLC and 29 cases of untreated SCLC (of which additional tumour biopsies after treatment with topo-IIalpha-directed drugs were available in ten cases) yielded the following results: NSCLC had significantly less topo-IIalpha than SCLC (P < 0.0001), as only 5 out of 27 NSCLC cases had > 5% positive cells compared with 28 out of 29 SCLC, and 0 out of 27 NSCLC had > 25% positive cells compared with 26 out of 29 SCLC. P-gp was detected in > 5% of cells in only 3 out of 27 NSCLC and in 6 out of 29 SCLC, and MRP in 5 out of 27 of NSCLC and 9 out of 29 SCLC. After treatment of patients with SCLC with either etoposide or teniposide, which are topo-IIalpha-directed drugs, there was an increase in MRP (P < 0.1) and P-gp (P < 0.05) positivity, while topo-IIalpha decreased (P < 0.05). In conclusion, the major difference between untreated NSCLC and SCLC was in topo-IIalpha content. In the small series of ten patients treated for SCLC, all three MDR phenotypes appeared to increase.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

The use of a stereological method to estimate the volume of Schistosoma mansoni granulomas: the effect of zinc deficiency.

A quantitative, stereological method to assess the volume of Schistosoma mansoni granulomas is presented and its use demonstrated in a study of zinc-deficient animals. Twenty-six mice were divided into two groups, and given zinc-deficient or control food. After 4 weeks, the mean weight gain and thymus size were lower in the mice on the zinc-deficient diet. Eight mice in each group were then each infected with 60 S. mansoni cercariae and these mice were killed 12 weeks post-infection. There were then more eggs in the livers of the zinc-deficient mice than in those of the infected controls (7400/female worm v. 3300/female worm; P = 0.02), probably because of impaired intestinal egg expulsion and impaired resorption of eggs in the deficient mice. The median proportion of each liver occupied by granulomas was 0.11 among the zinc-deficient mice and 0.09 among the pair-fed, control mice (P = 0.08), and the median liver volumes were 1400 and 1450 microliters (P = 0.49), respectively. The computed median volume of all the granulomas in each liver was 160 microliters in zinc-deficient and 132 microliters in control mice (P = 0.32), while median granuloma volume per egg was 8.6 and 8.7 nl (P = 0.93), respectively. In contrast to conventional methods, modern stereological methods can provide estimates of granuloma volume or granuloma volume fractions without any assumptions about the size, shape, orientation or distribution of the granulomas in the liver.

Animals↗

Expression of alpha and beta subunits of the integrin superfamily in articular cartilage from macroscopically normal and osteoarthritic human femoral heads.

OBJECTIVE: The objective of this study was to detail the topographical and zonal distribution of alpha and beta subunits of the integrin superfamily in normal and osteoarthritic cartilage. METHODS: Immunohistochemistry utilising antibodies towards alpha and beta subunits was performed on cryostat sections of human articular cartilage from macroscopically normal (n = 6) and osteoarthritic (n = 6) femoral heads. Samples of articular cartilage were obtained from 12 topographically distinct sites from each femoral head. Each section was divided into zones (superficial, middle, deep) and staining scores were recorded. RESULTS: Normal cartilage stained for integrin subunits alpha 1, alpha 5, alpha V, beta 1, beta 4, and beta 5, but not for alpha 2, alpha 3, alpha 4, alpha 6, beta 2, beta 3, and beta 6. Intact and non-intact residual cartilage from osteoarthritic femoral heads stained for alpha 1, alpha 2, alpha 5, alpha V, beta 1, beta 4, and beta 5. Staining was occasionally seen for alpha 4 and beta 2, but not for alpha 3, alpha 6, beta 3, and beta 6. There was no topographical variation in the staining for any of the subunits in either normal or osteoarthritic cartilage. The only subunit that displayed a zonal variation was alpha V; staining for this subunit was most pronounced in the superficial zone compared with the middle and deep zones. CONCLUSION: Chondrocytes in normal and osteoarthritic cartilage express the integrin subunits alpha 1, alpha 5, alpha V, beta 1, beta 4, and beta 5. Chondrocytes in osteoarthritic cartilage, in addition, express the alpha 2, alpha 4, and beta 2 subunits. The alpha v subunit is expressed by more chondrocytes in the superficial zone in comparison with cells in the deeper zones. None of the subunits display topographical variation in expression.

Aged↗

Modified cytokeratins expressed on the surface of carcinoma cells undergo endocytosis upon binding of human monoclonal antibody and its recombinant Fab fragment.

Previously, we have reported on successful imaging of colon, rectal, and pancreatic carcinomas in patients by using a radiolabeled all-human monoclonal antibody, COU-1, directed against modified cytokeratin. To further develop this antibody for use as an immunoconjugate, COU-1 was cloned by phage display selection and the human Fab fragment was expressed in bacteria. Analysis by confocal laser scanning microscopy demonstrated that COU-1 bound in a uniform punctate pattern to the surface of viable carcinoma cells stained at 4 degrees C, and binding increased significantly when cells were cultured on fibronectin, laminin, or collagen IV. In the case of fibronectin, COU-1 staining was particularly enhanced at intercellular junctions. When carcinoma cells were cultured with COU-1 at 37 degrees C for 6 hr, the antibody was found in large perinuclear vesicles and the punctate surface staining was significantly reduced. Similar results were obtained using intact IgM COU-1 and the recombinant Fab fragment. Immunohistological studies indicated that COU-1, in contrast to murine monoclonal antibodies against normal cytokeratin 8 and 18, could differentiate between malignant and normal colon epithelia, and between colon cancer metastasis in the liver and surrounding normal hepatocytes. Within biopsies of malignant tissue, COU-1 exhibited membrane-associated staining of proliferating cells, while resting cells had a filamentous pattern. Thus, modified cytokeratin at the surface of carcinoma cells may represent a new target for immunoconjugates and may explain the promising results of the phase I/II clinical study.

Amino Acid Sequence↗

[Clinical results after en block double lung transplantation with direct bronchial revascularization. The first three and a half years' experience in Denmark].

En-bloc double lung transplantation with tracheal anastomosis and direct revascularization of the bronchial arteries to the left internal mammary artery has been carried out in Denmark since June 1992. Forty-seven patients (32 with alfa-1 antitrypsin deficiency, 11 with chronic obstructive pulmonary disease, two with cystic fibrosis and two with primary pulmonary hypertension), 25 men and 22 women, average age 39 years (17-64 years), have received their first double-lung transplant with bronchial artery revascularization. Arteriography of the internal mammary artery and bronchial arteries was performed in 42 (89%) of the patients from 1-150 days after the operation. Successful bronchial artery revascularization was demonstrated arteriographically in 40 patients, in two patients the arteriography failed to show bronchial artery revascularization. Arteriography was not performed in five patients due to early complications and death. Bronchoscopy showed rapid, uncomplicated airway healing in 42 patients. Mucosal necrosis under the tracheal anastomosis was found in three patients, and severe obstructive endobronchial growth of the fungus Aspergillus fumigatus was diagnosed in the last two patients. The one- and two-year survival is 83% (Kaplan-Meier). Eleven patients are dead, five due to pulmonary causes and six due to extra-pulmonary causes. Pulmonary function became normal in nearly all surviving patients between three to six months after the transplantation. In conclusion, en-bloc double-lung transplantation with bronchial artery vascularization has shown good short-term results, and the one- and two-year survival gives hope that a successful bronchial artery revascularization will improve the long-term survival following lung transplantation.

Adolescent↗

Histologic/histochemical grading system for osteoarthritic articular cartilage: reproducibility and validity.

OBJECTIVE: To evaluate the reproducibility and validity of the histologic/histochemical grading system (HHGS) for osteoarthritis (OA) developed by Mankin and coworkers. METHODS: Sections of human articular cartilage from macroscopically normal and osteoarthritic femoral heads were graded by the HHGS (0-14 scale) twice by 3 observers, and intra- and interobserver reproducibility was determined. RESULTS: The exact intra- and interobserver reproducibilities ranged from 22% to 33% and from 13% to 25%, respectively. Intra- and interobserver variation ranged from -7 to 5. Although scores were generally higher in cartilage from osteoarthritic joints as compared with cartilage from normal joints, the difference reached statistical significance for only 2 observers. Additionally, the average scores from all observers were within the definitive range for OA (>5) in 30% of sections from normal joints, and in only 47% of sections from OA joints. CONCLUSION: The results indicate that the reproducibility and the validity of the HHGS are inadequate, and an improved histopathologic grading system for OA is needed.

Adult↗

Medium-term patency and anatomic changes after direct bronchial artery revascularization in lung and heart-lung transplantation with the internal thoracic artery conduit.

OBJECTIVE: Our purpose was to study the 2-year patency of direct bronchial artery revascularization in lung transplantation. We wanted to clarify whether the revascularized bronchial artery system is functional after 2 years, whether bronchial artery vascularity changes with time, and whether posttransplantation bronchial artery disease is arteriographically evident after 2 years. METHODS: Bronchial artery revascularization is performed by anastomosing the internal thoracic artery to as many bronchial artery orifices in the donor descending aorta as possible. Twenty-three patients surviving 2 years or more have had internal thoracic artery-bronchial arteriography performed 1 month and 2 years after transplantation. One-month and 2-year arteriograms have been compared. RESULTS: Two-year patency of the internal thoracic artery conduit was 100%. The appearance of the bronchial arteries was unchanged after 2 years in 11 patients. A unilateral or bilateral increase in vascularity was found in two and seven patients, respectively. In three patients new vessels, not visible on the first arteriogram, had appeared. In four patients one or more small vessels visible on the first arteriogram had disappeared on the second arteriogram. We have found no arteriographic signs of bronchial artery disease, such as stenosis of the bronchial arteries, and no arteriographic evidence of arteriosclerotic disease in the internal thoracic artery. CONCLUSION: The internal thoracic artery is an excellent conduit for bronchial artery revascularization, with a 2-year patency of 100% in 23 patients. Only minor changes in the bronchial arteriograms have been found.

Bronchial Arteries↗

CD44 expression is up-regulated in the deep zone of osteoarthritic cartilage from human femoral heads.

AIMS: The objective of this study was to detail the topographical and zonal distribution of the cell adhesion molecule CD44 in normal and osteoarthritic cartilage. METHODS AND RESULTS: Immunohistochemistry utilizing well characterized anti-CD44 antibodies (clones A3D8, Bric 235, 2C5) was performed on cryostat and paraffin sections of human articular cartilage from macroscopically normal (n = 18) and osteoarthritic (n = 11) femoral heads. Samples for cryostat sections were obtained from 12 topographically different sites. Sections were divided into zones (superficial, middle, deep) and the CD44 staining scored. Chondrocytes in normal articular cartilage and cartilage from osteoarthritic femoral heads stained positive for CD44 in both cryostat and paraffin sections. Normal cartilage showed a significant decrease in CD44 staining in the deep zone as compared to the superficial zone (P < 0.05). However, cryostat sections of residual cartilage from osteoarthritic femoral heads showed increased CD44 staining in the deep zone as compared to normal articular cartilage. The CD44 staining showed no topographical variation in either the normal cartilage or the osteoarthritic residual cartilage. CONCLUSIONS: CD44 expression displays a distinct zonal variation in normal articular cartilage which is lost in osteoarthritic cartilage due to an up-regulated expression in the deep zone. CD44 expression does not exhibit topographical variation.

Aged↗

Interaction between proximal tubular cells and allogenic leukocytes mediated by interleukin-1 and prostaglandin.

The present study investigates the interaction between proximal tubular cells (TC) and peripheral blood mononuclear cells (PBMC) with respect to interleukin-1 (Il-1) and prostaglandin E2 (PGE2) in co-culture experiments. Stimulator cells were proliferating human TC propagated in culture and characterized by expression of ICAM-1 and MHC-class II in response to cytokines. The presence of TC reduced spontaneous growth of PBMC by 42%, with a concomittant 50% reduction of Il-1 release and a 6-fold increase of PGE2 release in the growth medium. The growth reduction was partly counteracted by addition of II-1 and indomethacin. Addition of II-1 stimulated the PGE2 release from PBMC and TC by about 75% and 500%, respectively. Growth of TC cultured alone was inhibited by II-1 in a dose-dependent manner, with maximum effect at 10 U/ml, whereas PGE2 showed no effect. These results suggest that TC have the capacity to interact with PBMC, and this potentially tissue-protective mechanism induced by TC may have clinical implications.

Cell Division↗

Dynamic autoregulation and renal injury in Dahl rats.

The Dahl salt-sensitive (Dahl S) rat develops hypertension and renal injuries when challenged with a high salt diet and has been considered to be a model of chronic renal failure. Renal injuries appear very early in life compared with the spontaneously hypertensive rat (SHR). During the course of hypertension, a gradual impairment of autoregulatory control of renal blood flow might expose the glomerular circulation to periods of elevated pressure, resulting in renal injuries in Dahl S rats. Dynamic autoregulatory capacity was assessed in Dahl S and Dahl salt-resistant (Dahl R) rats, SHR, and Sprague-Dawley rats by inducing broad-band fluctuations in the arterial blood pressure and simultaneously measuring renal blood flow. Dynamic autoregulation was estimated by the transfer function using blood pressure as the input and renal blood flow as the output. Renal morphological injuries were evaluated in Dahl S rats and SHR and were scored semiquantitatively. Dynamic autoregulation was efficient and comparable in the low-frequency range (<0.015 Hz) in Dahl R rats, SHR, and Sprague-Dawley rats. The response in Dahl S rats depended strongly on the initiation time of the high salt diet. Autoregulation was preserved during a low salt diet and in rats exposed to a late-onset hypertension of short duration, only partly preserved if the late-onset hypertension was of a longer duration, and abolished in early-onset hypertension. All Dahl S rats on a high salt diet showed severe morphological changes in the kidney. In conclusion, autoregulatory capacity in the kidney of Dahl S rats is gradually impaired when rats are rendered hypertensive with a high salt diet. Renal morphological injuries develop before loss of dynamic autoregulation. Impaired autoregulation appears to be the result, not the cause, of the process that ultimately leads to renal failure in the Dahl S rat.

Animals↗