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PubMed1978

Studies on a phospholipase B from Penicillium notatum. Substrate specificity.

1. The action of a highly purified phospholipase B from Penicillium notatum on 1-O-alk-1'-enyl-2-acyl-, 1-O-alkyl-2-acyl-, 1,2-diacyl-, 1-acyl- and 2-acyl-sn-glycero-3-phosphocholine, monoacyl-, diacyl- and triacylglycerols, cholesteryl oleate and p-nitrophenyl acetate was studied. 2. The hydrolysis products of the monoethermonoacylglycerophospholipids were identified as fatty acids, 1-O-alk-1'-enyl-sn-glycero-3-phosphocholine and 1-O-alkyl-sn-glycero-3-phosphocholine. The hydrolysis rates were in the following order: 1,2-diacyl-sn-glycero-3-phosphocholine greater than 1-O-alk-1'-enyl-2-acyl-sn-glycero-3-phosphocholine greater than 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine. 3. 1-Acyl-sn-glycero-3-phosphocholine was hydrolyzed about 15 times faster than 2-acyl-sn-glycero-3-phosphocholine. 4. Monoacylglycerols were hydrolyzed at the optimal pH 4.0, but diacyl- and triacylglycerols were not hydrolyzed at various pH values between 4.0 and 9.0. 5. Cholesteryl oleate and p-nitrophenyl acetate were not hydrolyzed.

Acetates↗
PubMed1993

Identification of a lysophospholipase C that may be responsible for the biosynthesis of choline plasmalogens by Madin-Darby canine kidney cells.

The biosynthesis of choline plasmalogens was investigated in Madin-Darby canine kidney cells to determine the source of the vinyl ether linkage. 1-O-[3H] Alk-1'-enyl-2-lyso-sn-glycero-3-phosphoethanolamine was a better precursor than 1-O-[3H]alkyl-2-lyso-sn-glycero-3-phosphocholine for the synthesis of 1-O-[3H]alk-1'-enyl-2-acyl-sn-glycero-3-phosphocholine; this suggests that the vinyl ether linkage in choline phosphoglycerides originates from ethanolamine plasmalogens. The contribution of N-methylation and base exchange enzymes to choline plasmalogen biosynthesis was assessed using 1-O-[3H]alkenyl-2-lyso-sn-glycero-3-[32P]phosphoethanolamine. While 1-O-[3H]alkenyl-2-acyl-sn-glycero-3-phosphocholine was formed from this precursor, the 32P was lost indicating that N-methylation and base exchange enzymes do not contribute significantly to the synthesis of choline plasmalogens. The conversion of a phosphono analog of 1-O-[3H]alkyl-2-lyso-sn-glycero-3-phosphoethanolamine, which is resistant to phospholipase D hydrolysis, to 1-O-[3H]alkenyl-2-acyl-sn-glycero-2-phosphocholine was observed demonstrating that phospholipase D is not required for choline plasmalogen biosynthesis. A Mg(2+)-dependent lysophospholipase C activity was detected in microsomes that actively hydrolyzed ether-linked lysophosphoglycerides as well as the lysophosphono analog. To assess the role of lysophospholipase C in shuttling 1-O-alk-1'-enyl-sn-glycerol (alkenylglycerol) from ethanolamine plasmalogens to choline plasmalogens, cells prelabeled with 1-O-[3H]alkenyl-2-lyso-sn-glycero-3-phosphoethanolamine were treated with 12-O-tetradecanoylphorbol-13-acetate. This resulted in the rapid deacylation of 1-O-[3H]alkenyl-2-acyl-sn-glycero-3-phosphoethanolamine to 1-O-[3H]alkenyl-2-lyso-sn-glycero-3-phosphoethanolamine and the subsequent generation of 1-O-[3H]alkenylglycerol. A concomitant 2-3-fold increase in 1-O-[3H]alkenyl-2-acyl-sn-glycero-3-phosphocholine was observed. These studies suggest that the alkenyl linkage in choline phosphoglycerides may originate from 1-O-alkenyl-2-lyso-sn-glycero-3-phosphoethanolamine through an enzymatic pathway involving lysophospholipase C to generate alkenylglycerol that is subsequently converted to choline plasmalogens.

Animals↗
PubMed1996

[Effect of sn-2-acetylated analogs of phosphatidalic acid on release and aggregation of human platelets].

The influence of sn-2-acetylated analogs of phosphatidalic acid on the release reaction and aggregation of human platelets has been studied. It was found that phosphate-, phosphorylethanolamine- or phosphorylcholine-containing sn-2-acetylated analogs of phosphatidalic acid do not stimulate human platelet aggregation but inhibit the platelet-activating factor (PAF) induced platelet aggregation. The phosphatidalic acid analog, I-O-alk-1'-enyl-2-acetyl-sn-glycero-3-phosphate, stimulates human platelet release reaction measured by acridine orange release from human platelets in the same extent as does PAF. Other phosphatidalic acid analogs (1-O-alk-1'-enyl-2-acetyl-sn-glycero-3-phosphoethanolamine, 1-O-alk-1'-enyl-2-acetyl-sn-glycero-3-phosphocholine) inhibit acridine orange exocytosis from PAF-stimulated human platelets. It is concluded that the activity of sn-2-acetylated phosphatidalic acid analogs towards human platelets differs from those of corresponding ester and ether phosphatidalic acid analogs and, consequently, they may have another functional significance in the processes of platelet activation.

Acetylation↗
PubMed1996

A preliminary clinical report of automated lamellar keratoplasty with excimer laser keratectomy for correction of high myopia.

OBJECTIVE: To evaluate the therapeutic effects of automated lamellar keratoplasty combined with excimer laser keratectomy (ALK-E) for the correction of high myopia. METHODS: With the automated corneal shaper and Keracor 116 excimer laser, ALK-E was performed on 47 eyes of 40 cases with myopia of -8.00 to -30.00 diopters (mean, -14.55D +/- 5.14D) and the highest astigmatism 7.00D. Postoperatively, all patients have been followed up for at least 3 months. RESULTS: The refractive status and visual acuity tend to be stable about 1 month postoperatively. For myopes below -15.00D, the mean refraction was -1.33D +/- 1.05D at 3 months, and 66.7% achieved their preoperative best corrected visual acuity. For myopes over -15.00D, the mean refraction was -2.42D +/- 1.84D at 3 months and 52.9% achieved their preoperative best corrected visual acuity. CONCLUSIONS: The early therapeutic results of ALK-E for treatment of high myopia are very promising. However, the operative techniques, parameters and remote therapeutic results remain to be further studied.

Adolescent↗
PubMed1997

The use of fluorescent in situ hybridization for detection of the t(2;5)(p23;q35) translocation in anaplastic large-cell lymphoma.

BACKGROUND: Anaplastic large-cell lymphoma (ALCL) is a recently recognized, distinctive type of non-Hodgkin's lymphoma characterized by anaplastic large-cell cytology and expression of a member of the TNF-receptor family CD30. A characteristic chromosomal translocation has been identified in ALCL of T-or null-cell lineage which juxtaposes a novel tyrosine kinase (anaplastic lymphoma kinase, ALK) located at 2p23 with the nucleophosmin gene (NPM) at 5q35. A chimeric mRNA transcript is produced, and the translocation results in constitutive expression of a truncated form of the ALK protein, p80. There is controversy concerning whether or not the translocation occurs in Hodgkin's disease. The aim of this study was to develop a methodology for fluorescent in situ hybridization (FISH) to detect the t(2;5)(p23;q35), and to compare the results with conventional cytogenetics, reverse-transcriptase PCR and immunostaining for the p80 protein. PATIENTS AND METHODS: Twenty-five cases of malignant lymphoma (11 ALCL and 14 HD) were studied. Immunohistochemistry was performed to confirm the diagnosis and for analysis of p80 expression. Conventional cytogenetics were analyzed on G-banded metaphase spreads. FISH was performed using whole chromosome paints for chromosomes 2 and 5 on metaphase spreads and YAC probes for interphase nuclei. Reverse-transcriptase PCR using primers for ALK and NPM was used to amplify the translocation breakpoint in extracted mRNA. RESULTS: Among 11 cases of ALCL examined by FISH, the translocation was detected in 4. Two of these cases also had RT-PCR and p80 staining performed, with positive results. Among 7 cases where the t(2;5) was not detected by FISH, 3 cases were examined by RT-PCR with negative results and 4 cases by p80 staining, also negative. The RT-PCR was negative in all 14 cases of Hodgkin's disease, 4 of which were also examined by FISH and found to be negative. CONCLUSION: Fluorescent in situ hybridization is a useful method for detection of the t(2;5)(p23;q35) in anaplastic large-cell lymphoma. The results concur with those of RT-PCR for the chimeric transcript and immunostaining for the p80 protein. The frequency with which the translocation was found was 36% in this small series, and no evidence of the translocation was found in cases of Hodgkin's disease.

Antigens, Neoplasm↗
PubMed1997

MNDA binds NPM/B23 and the NPM-MLF1 chimera generated by the t(3;5) associated with myelodysplastic syndrome and acute myeloid leukemia.

The myeloid cell nuclear differentiation antigen (MNDA) is a nuclear protein expressed specifically in developing cells of the human myelomonocytic lineage, including the end-stage monocytes/macrophages and granulocytes. Nuclear localization, lineage- and stage-specific expression, association with chromatin, and regulation by interferon alpha indicate that this protein is involved in regulating gene expression uniquely associated with the differentiation process and/or function of the monocyte/macrophage. MNDA does not bind specific DNA sequences, but rather a set of nuclear proteins that includes nucleolin (C23). Both in vitro binding assays and co-immunoprecipitation were used to demonstrate that MNDA also binds protein B23 (nucleophosmin/NPM). Three reciprocal chromosome translocations found in certain cases of leukemia/lymphoma involve fusions with the NPM/B23 gene, t(5;17) NPM-RARalpha, t(2;5) NPM-ALK, and the t(3;5) NPM-MLF1. In the current study, MNDA was not able to bind the NPM-ALK chimera originating from the t(2;5) and containing residues 1-117 of NPM. However, MNDA did bind the NPM-MLF1 product of the t(3;5) that contains the N-terminal 175 residues of NPM. The additional 58 amino acids (amino acids 117-175) of the NPM sequence that are contained in the product of the NPM-MLF1 fusion gene relative to the product of the NPM-ALK fusion appear responsible for MNDA binding. This additional NPM sequence contains a nuclear localization signal and clusters of acidic residues believed to bind nuclear localization signals of other proteins. Whereas NPM and nucleolin are primarily localized within the nucleolus, MNDA is distributed throughout the nucleus including the nucleolus, suggesting that additional interactions define overall MNDA localization.

Acute Disease↗
PubMedDate not supplied

Intraperitoneal pulse calcitriol in the treatment of secondary hyperparathyroidism in patients on continuous ambulatory peritoneal dialysis.

OBJECTIVE: To determine whether it is practicable to use intraperitoneal calcitriol to treat continuous ambulatory peritoneal dialysis (CAPD) patients with secondary hyperparathyroidism and whether this form of therapy is effective and safe. DESIGN: A prospective, nonrandomized study. SETTING: Division of Nephrology, Tertiary Hospital. METHOD: Eight patients from our CAPD population were recruited (5 F, 3 M), aged 24 to 63 years (mean 38.9 +/- 7.6 yr). They had been on CAPD for 8 to 84 months (mean 47.6 +/- 24.6 months). All the patients had bone biopsy-proven secondary hyperparathyroidism with 2 patients showing mixed lesions. The CAPD system was changed to the twin-bag system (Ultrabag, Baxter Healthcare, McGaw Park, IL, U.S.A.) in all 8 patients, who were taught to inject the calcitriol directly through the short transfer set and the Tenckhoff catheter into the peritoneal cavity, twice per week before bedtime. Calcium carbonate or calcium acetate was used as the main phosphate binder. Intact parathyroid hormone level (iPTH), serum ionized calcium (iCa), serum phosphate, and serum total alkaline phosphatase (alk. phos.) levels were measured at baseline and then every 4 weeks. The mean duration of follow-up was 10.5 months +/- 1.9 months. RESULTS: There was a significant drop of iPTH level from the pretreatment level of 100.6 +/- 35.8 pmol/L to a level of 63.8 +/- 48.7 pmol/L at 24 weeks (p = 0.036). The lowest level of iPTH attained was 43.4 +/- 27.0 pmol/L at 48 weeks. Serum total alk. phos. also dropped from 232.4 +/- 83.3 IU/L pretreatment to 147.9 +/- 52.0 IU/L at 24 weeks (p = 0.017). The decrease in alk. phos. level paralleled the decrease in iPTH level. The mean serum iCa level did not show any significant rise throughout the study period. The maximum dose of calcitriol used was 6.6 +/- 1.5 microg/week and the average dose of calcitriol was 5.4 +/- 1.2 microg/week. One patient did not respond satisfactorily and she subsequently had a parathyroidectomy. Two episodes of peritonitis occurred during the study period, giving a peritonitis rate of one episode per 42 patient-months. There was no significant change in the urea clearance tests or the peritoneal equilibration tests before and after the study. CONCLUSION: Intraperitoneal calcitriol is practicable, effective, and safe in the treatment of secondary hyperparathyroidism in CAPD patients.

Adult↗
PubMed1996

Dexamethasone regulation of marrow stromal-derived osteoblastic cells.

The clonal subtypes of cells in the osteogenic family represented by fibroblastoid MBA-15.33, preosteoblast MBA-15.4, and mature osteoblastic MBA-15.6 cells were used to study the effects of glucocorticoid (dexamethasone). The role of dexamethasone was monitored on cell attachment when plated on various protein substrata (BSA, collagen 1, and Matrigel). A 24 h exposure of the cells to 10(-6) M or 10(-7) M dexamethasone differential affects their attachment preference. MBA-15.33 and MBA-15.4 cells increased their attachment capability on collagen 1, while MBA-15.6 cells' attachment was inhibited. Pretreatment with (10(-6) M) dexamethasone caused an increase in attachment on Matrigel by MBA-15.33 cells and to less extent by MBA-15.4 cells. Additionally, measurements of two enzymatic activities were monitored; one is alkaline phosphatase (ALK-P), and the second is neutral endopeptidase (CD10/NEP). MBA-15.33, MBA-15.4, and MBA-15.6 cells were exposed to dexamethasone or to various growth factors (bone morphogenic protein (BMP-2 and BMP-3), TGF beta, and IGF-1). In some experiments, pretreatment of cells by dexamethasone was followed by exposure to the growth factors. The cells' challenged cellular responses were not uniform and revealed a differential pattern when their ALK-P and CD10/NEP enzymatic activities were measured.

Animals↗
PubMed1990

Analysis of prognostic factors in disseminated prostatic cancer. An update. Dutch Southeastern Urological Cooperative Group.

A statistical analysis of prognostic factors in 175 patients with hormonally treated disseminated prostatic cancer was done. The prognostic significance of performance status (PS), hemoglobin (Hb), alkaline phosphatase (Alk P), and testosterone was assessed with a univariate analysis. The authors did not find significant prognostic value in age, tumor size or grade, prostatic acid phosphatase, and prostate-specific antigen in these patients. In a multivariate logistic model (Cox regression), PS, Hb, and Alk P were found useful for dividing patients into prognostic groups. The prognosis for high-risk patients on standard hormonal treatment was very poor. The authors concluded that research on prognostic factors is useful and permits a division of patients into risk groups that makes choice of treatment more accurate. The use of new treatment combinations as a start treatment is appropriate for high-risk patients with disseminated prostatic cancer.

Acid Phosphatase↗
PubMed2001

Interaction between topography and coating in the formation of bone nodules in culture for hydroxyapatite- and titanium-coated micromachined surfaces.

Rat osteoblast cultures were maintained from 24 h to 6 weeks on hydroxyapatite (HA)- or titanium (Ti)-coated smooth and micromachined grooved substrata in medium supplemented with L-ascorbic acid-2-phosphate and beta-glycerophosphate to promote mineralization. The HA coatings, approximately 1 microm thick, were characterized using X-ray diffraction, surface roughness, and scanning electron microscopy (SEM). Osteoblasts elongated, aligned, and moved in the direction of the grooves on both Ti and HA grooved surfaces. HA surfaces produced significantly more bone-like nodules than Ti surfaces. All grooved substrata produced significantly more nodules than smooth surfaces. These results are consistent with the hypothesis that substrata can increase osteogenesis by formation of an appropriate microenvironment. There was also a statistically significant interaction between topography and chemistry in the formation of mineralized nodules. A strong correlation (r = 0.958) between alkaline phosphatase (Alk-P) at 2 weeks and nodule counts at 6 weeks was observed, suggesting that Alk-P may possibly be used as a leading indicator of osteogenesis on microfabricated surfaces. The results of this study indicate that surface topography and chemistry can affect osteogenesis, and that interactions between chemistry and topography can occur.

Animals↗
PubMed2001

Systemic anaplastic large-cell lymphoma: results from the non-Hodgkin's lymphoma classification project.

Anaplastic large-cell lymphoma (ALCL) is a heterogeneous process that may have a T-cell, B-cell, or indeterminant (null) phenotype and which may or may not express the anaplastic lymphoma kinase (ALK) oncoprotein. Because the clinical significance of these variants of ALCL is unclear, we evaluated the cases of ALCL-T/null and ALCL-B identified in the Non-Hodgkin's Lymphoma Classification Project. We evaluated 1,378 cases of non-Hodgkin's lymphoma (NHL), and a consensus diagnosis of ALCL-T/null was made in 33 patients (2.4%) with a diagnostic accuracy of 85%. Compared to 96 patients with other forms of peripheral T-cell lymphoma (PTCL), those with ALCL-T/null were significantly younger, less likely to have advanced-stage disease or bone marrow involvement, more likely to have a low International Prognostic Index score, and had a significantly better survival. Among those with ALCL-T/null, there were no significant differences in the clinical features or survival on the basis of ALK expression. A consensus diagnosis of ALCL-B was made in 15 patients (1.1%), and the diagnostic accuracy was 67%. However, compared to 366 patients with other forms of diffuse large B-cell lymphoma (DLBCL), those with ALCL-B were no different with regard to clinical features or survival. We conclude that patients with ALCL-T/null have favorable prognostic features and excellent survival and should be separated from those with other forms of PTCL for prognostic and therapeutic purposes. In contrast, patients with ALCL-B appear to be similar to those with other forms of DLBCL.

Adult↗
PubMed2005

Cost comparison of genetic and clinical screening in families with hereditary hemorrhagic telangiectasia.

Endoglin (ENG) and ALK-1 mutations cause hereditary hemorrhagic telangiecstasia (HHT), an autosomal dominant disorder leading to vascular dysplasia in the form of mucocutaneous telangiectasia and visceral arteriovenous malformations (AVMs). We proposed to compare two alternative strategies for management of HHT: screening HHT families with molecular diagnostic tests followed by targeted clinical screening versus conventional clinical screening. A decision analytic model was constructed to compare screening strategies for a hypothetical HHT family. The family consists of 1 index case and 13 relatives. The clinical screening protocol in use at the Canadian HHT Center in Toronto was assumed to be the standard of care. Unit costs for clinical screening (in Canadian dollars) were obtained from the 2003 Ontario Health Insurance Schedule of Benefits. Genetic screening costs were estimated for quantitative multiplex PCR and sequencing of Endoglin (ENG) and ALK-1 genes, as performed at HHT Solutions, Toronto. The genetic screening strategy resulted in a net cost of $4,060 per individual versus $5,975 for the clinical screening strategy. The genetic screening strategy would save $1,915 per family member or $26,810 saved per family. Sensitivity analyses revealed that the genetic screening strategy was cost saving over all plausible ranges of input variables for all hypothetical families tested. We concluded that a genetic screening strategy with targeted clinical screening is more economically attractive than conventional clinical screening and results in a reduction in the number of clinical tests for family members who do not have HHT.

Adolescent↗
PubMed2005

Towards supramolecular fixation of NOX gases: encapsulated reagents for nitrosation.

The use of simple calix[4]arenes for chemical conversion of NO2/N2O4 gases is demonstrated in solution and in the solid state. Upon reacting with these gases, calixarenes 1 encapsulate nitrosonium (NO+) cations within their cavities with the formation of stable calixarene-NO+ complexes 2. These complexes act as encapsulated nitrosating reagents; cavity effects control their reactivity and selectivity. Complexes 2 were effectively used for nitrosation of secondary amides 5, including chiral derivatives. Unique size-shape selectivity was observed, allowing for exclusive nitrosation of less crowded N-Me amides 5 a-e (up to 95 % yields). Bulkier N-Alk (Alk>Me) substrates 5 did not react due to the hindered approach to the encapsulated NO+ reagents. Robust, silica gel based calixarene material 3 was prepared, which reversibly traps NO2/N2O4 with the formation of NO+-storing silica gel 4. With material 4, similar size-shape selectivity was observed for nitrosation. The N-Me-N-nitroso derivatives 6 d,e were obtained with approximately 30 % yields, while bulkier amides were nitrosated with much lower yields (<8 %). Enantiomerically pure encapsulating reagent 2 d was tested for nitrosation of racemic amide 5 t, showing modest but reproducible stereoselectivity and approximately 15 % ee. Given high affinity to NO+ species, which can be generated by a number of NOX gases, these supramolecular reagents and materials may be useful for NOX entrapment and separation in the environment and biomedical areas.

Amides↗
PubMed2005

Ribozyme-targeting reveals the rate-limiting role of pleiotrophin in glioblastoma.

Glioblastomas (GBMs) are the most frequent malignant brain tumors with very limited treatment options and nearly all GBM patients dying within 1 year. Pleiotrophin (PTN, HB-GAM, HBNF, OSF-1) is a secreted growth factor that shows mitogenic, chemotactic and transforming activity. While PTN expression is tightly regulated during embryogenesis and very limited in normal adult tissues, a marked PTN upregulation is seen in tumors including glioblastomas. Targeting of the PTN receptors, ALK and RPTP-zeta, indicates a contribution of PTN-activated signaling pathways in glioblastomas. However, the relevance of PTN expression itself is unknown especially since, besides PTN, at least one more growth factor, midkine (MK), signals through ALK and is expressed in glioblastoma. Here we demonstrate the biologic relevance of PTN in 2 glioblastoma cell lines in vitro and in vivo. We show that stable ribozyme-targeting leads to a robust reduction of PTN mRNA and protein levels. This results in decreased cell proliferation, cell migration and soft agar colony formation in vitro. Comparing clonal ribozyme-transfected cells with different residual PTN levels, we establish a PTN gene-dose effect of glioblastoma cell proliferation. In a subcutaneous tumor xenograft mouse model, in vivo growth is markedly reduced upon PTN depletion, which is paralleled by decreased PTN serum levels. Furthermore, the immunohistochemical analysis of the tumors shows reduced angiogenesis in PTN-depleted tumors. We conclude that PTN is a rate-limiting growth factor in glioblastoma. Since PTN is overexpressed in glioblastomas but rarely found in normal tissue, PTN may represent an attractive therapeutic target.

Animals↗
PubMed2006

Expression and regulation of CReMM, a chromodomain helicase-DNA-binding (CHD), in marrow stroma derived osteoprogenitors.

This study follows the expression of CReMM, a new CHD family member, in osteoprogenitors. CReMM expression was analyzed in primary cultured mesnchymal cells from rat and human. Analysis in ex vivo cultured marrow stromal cells (MSC) from rats revealed higher level of CReMM in cells from young (3 months), when compared to cells from old (15 months) rats. CReMM level was higher in human MSC then in mature trabecular bone cells (TBC). Within the MSC population, osteogenic clones showed higher levels of CReMM then non-osteogenic ones. We used bone marrow derived osteogenic cell line (MBA-15) to elaborate on the regulation of CReMM expression in correlation with cell proliferation and co-expression with alkaline phosphatase (ALK). CReMM is highly expressed in proliferating cells and is inversely related to expression of ALK. MBA-15 cells were challenged with dexamethasone (Dex) or 17beta-estradiol and quantification of CReMM at the protein (ELISA) and mRNA (RT-PCR) levels had shown that Dex upregulated CReMM levels. Since CReMM is regulated by Dex, we analyzed the interaction of CReMM with the glucocorticoid receptor (GR), which mediates Dex action. Co-immunopercipitation (Co-IP) demonstrated an association between CReMM and GR. In summary, CReMM is a CHD protein expressed by osteoprogenitors, and we suggest it plays a role in mediating transcriptional response to hormones that coordinate osteoblast function.

Animals↗
PubMed2006

Acyl-CoA oxidase activity from Beauveria bassiana, an entomopathogenic fungus.

Beauveria bassiana produces acyl-Co oxidase (ACO) in the P(20000 g) fraction of glucose and alkane-grown cultures that catalyze the oxidation of acyl-CoAs of different chain length. The activity was measured indirectly over the formation of H2O2 via the oxidative-coupled assay system. ACO activity was assessed spectrophotometrically in the P(20000 g) fraction of glucose-grown (FS0) and n-alkane grown cultures (FS(alk)), employing acyl-CoAs of 16 to 24 carbons as substrates. A significant increment in the activity was observed in FS(alk) as compared to that of controls (FS0) in all conditions tested. Tetracosane-grown cultures showed the highest activity with lignoceroyl-CoA. The reaction conditions were optimized employing lignoceroyl-CoA as substrate. A variable lag phase was observed when the activity was measured as a function of time. In the presence of 3-amino-1,2,4-triazole (AT) to prevent H2O2 consumption by endogenous catalase, the lag phase became shorter and disappeared when AT concentrations were raised from 40 to 200 mM, thus enhancing acyl-CoA oxidation. Enzyme activity reached its maximal value in the presence of 240 microg peroxidase, 0.08% Triton X-100 and 36 microM bovine serum albumin. The apparent Km using lignoceroyl as substrate was estimated 2.5 microM. ACO showed high activity and stability between 30 and 40 degrees C, as well as between 7.0 and 9.0 pH, for 120 min, being 7.0 the optimum pH.

Acyl-CoA Oxidase↗
PubMed1980

Liver scanning in newly diagnosed breast carcinoma.

The records of 111 consecutive cases of carcinoma of the breast were reviewed and yielded 66 patients who had undergone liver scanning and determination of the serum alkaline phosphatase (alk p'tase), lactic dehydrogenase (LDH), and serum glutamic oxaloacetic transaminase (SGOT). Four of the 66 patients (6.1%) had a focal defect on the liver scan. Two of those two had normal enzymes and two had at least one enzyme elevation. The two with normal enzymes have enjoyed prolonged disease-free survivals. The two with an elevated enzyme have both expired due to metastatic disease. No correlation could be found between the number of involved axillary nodes at surgery and the risk of having a focal defect on the liver scan. This study suggests that routine radionuclide liver scanning in the perimastectomy period does not add useful information in the face of normal alk p'tase, LSH, and SGOT.

Alkaline Phosphatase↗
PubMed2001

Prognostic significance of p80 and visceral involvement in childhood CD30 anaplastic large cell lymphoma (ALCL).

BACKGROUND: Between 1976 and 1998, CD30+anaplastic large cell lymphoma (ALCL) was diagnosed in 44 children (28 males, 16 females, age range 2.7-16.1 years, median 10). Up to 1993, 32 such children were treated according to a common chemotherapeutic protocol that stratified patients according to stage, without considering presenting features. Thereafter, patients presenting with visceral (lung, spleen, liver, gastro-intestinal tract) or mediastinal involvement were assigned to a high-risk treatment protocol with induction intensification. The generation of these two risk-groups was the result of a retrospective analysis of clinical risk factors for therapy failure as previously reported [Massimino M, Gasparini M, Giardini R, Ann Oncol 1995;6:915-920]. Considering the whole cohort of patients divided into group A-21/22 evaluable patients with visceral/mediastinal involvement, and group B-22 evaluable patients, with other ALCL location-disease-free survival (DFS) and survival (S) at 5 years were 57 and 58% for group A, and 83 and 100% (94% at 6 years) for group B, respectively. PROCEDURE: We tested 15/21 cases of group A, and 18/22 of group B for p80 immunoreactivity in order to investigate a possible correlation between ALCL locations and NPM-ALK expression. RESULTS: Thirteen of 15 specimens in group A and 17/18 in group B were positive for p80. CONCLUSIONS: It is impossible to conclude anything about p80 positivity based on a series of 33/44 patients with childhood ALCL, neither about over-all prognosis nor about the role of visceral involvement. In adults, NPM-ALK protein expression is a favourable prognostic factor. Med Pediatr Oncol 2001;37:97-102.

Adolescent↗