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Biomedical subjects

A M Scanu

Publications and source records attributed to A M Scanu.

At least 37 records · Page 2Linked to original sources
PubMed1998

Apolipoprotein(a) binds via its C-terminal domain to the protein core of the proteoglycan decorin. Implications for the retention of lipoprotein(a) in atherosclerotic lesions.

Although it is known that lipoprotein(a) (Lp(a)) binds to proteoglycans, the mechanism for this binding has not been fully elucidated. In order to shed light on this subject, we examined the interactions of decorin, a proteoglycan with a well defined protein core and a single glycosaminoglycan (GAG) chain, with Lp(a) and derivatives, namely Lp(a) deprived of apo(a), or Lp(a-), free apo(a), and the two main proteolytic fragments, F1 and F2. By circular dichroism criteria, the decorin preparations used had the same secondary structure as that previously reported for native decorin. Authentic low density lipoprotein from the same human donor was used as a control. In a solid phase system, Lp(a-)and low density lipoprotein bound to decorin in a comparable manner. This binding required Ca2+/Mg2+ ions, was lysine-mediated, and was markedly decreased in the presence of GAG-depleted decorin, suggesting the ionic nature of the interaction likely involving apoB100 and the GAG component of decorin. Free apo(a) also bound to decorin; however, the binding was neither cation-dependent nor lysine-mediated, unaffected by sialic acid depletion of apo(a), and markedly decreased when either reduced and alkylated apo(a) or reduced and alkylated decorin was used in the assay. Of note, the binding of apo(a) was unaffected when it was incubated with a spectrally native decorin that had been renatured from either 4 M guanidine hydrochloride by extensive dialysis or cooled from 65 to 25 degrees C. On the other hand, the binding significantly increased when decorin was depleted of GAGs, which by themselves had no affinity for apo(a). The binding of apo(a) to the decorin protein core was also elicited by the C-terminal domain of apo(a), and it was favored by high NaCl concentrations, 1 to 2 M. No binding was exhibited by the N-terminal domain accounting for the lack of effect of apo(a) size polymorphism on the binding. In the case of whole Lp(a), the binding to immobilized decorin was mostly GAG-dependent and ionic in nature. A minor contribution by apo(a) was detected when GAG-depleted decorin was used in the assay. Our results indicate that the binding of Lp(a) to decorin involves interactions both electrostatic (apoB100-GAG) and hydrophobic (apo(a)-decorin protein core), and that the binding of apo(a) requires decorin protein core to be in its native state.

Animals↗
PubMed1998

Tardive presentation of congenital cystic adenomatoid malformation of the lung.

Congenital cystic adenomatoid malformation (CCAM) of the lung is a neonatal disease not often found after the first year of life and extremely rare in adults. Three cases of CCAM, one in a ten-year-old girl and two in adults, are reported. An understanding of this disease is important because, although relatively rare, it is one of the most frequent causes of neonatal respiratory distress. Greater awareness of the condition and its early detection would also reduce the number of cases found at a later age.

Adolescent↗
PubMed1998

Congenital carotid-jugular fistula in an elderly patient.

Carotid-jugular arteriovenous fistulas (AVF) are extremely rare with only 20 cases reported in the literature up to December 1996. The case of a 74-year-old man (the oldest reported in the literature) with abnormal communications between the external carotid artery and the internal jugular vein is reported. The condition was treated by platinum coil embolization via catheterization and by repeated operations to ligate the branches of the external carotid artery, besides removal of the tissue containing the fistula and ligation of the external carotid at its origin. The result was incomplete because about 6 months after the last operation the patient showed the same symptoms, although in a milder form. Until standard treatment is established, the appropriate technique should be decided on a patient-to-patient basis.

Aged↗
PubMed1997

Polymorphonuclear cells isolated from human peripheral blood cleave lipoprotein(a) and apolipoprotein(a) at multiple interkringle sites via the enzyme elastase. Generation of mini-Lp(a) particles and apo(a) fragments.

Incubation of polymorphonuclear cells (PMN), isolated from human peripheral blood, with either lipoprotein(a) (Lp(a)) or free apolipoprotein(a) (apo(a)), derived from the parent Lp(a), caused in both cases a multisite fragmentation of apo(a) inhibited by methoxysuccinyl-Ala-Ala-Pro-Val-CH2Cl, a specific elastase inhibitor. The major cut site was at the interkringle region between apo(a) kringles IV-4 and IV-5 (Ile3520-Leu3521). The other cleavages were between kringles IV-7 and IV-8 (Thr3846-Leu3847) and between kringles IV-10 and V (Ile4196-Gln4197). The elastase-induced fragmentation of apo(a) was the same whether free or as a member of Lp(a), indicating that the disulfide bond between apo(a) and the apoB100 component of Lp(a) did not hinder the elastase action. Lp(a) fragments containing kringle IV-9 retained the linkage to apoB100 via the disulfide bond, forming mini-Lp(a) particles in which the size of apo(a) varied according to the size of the fragments produced by the elastase digestion. The proteolytic fragmentation was unaffected by apo(a) size polymorphism within the range examined. PMN elastase also caused a partial proteolysis of apoB100 whether as a component of Lp(a), Lp(a) freed of apo(a), or authentic low density lipoprotein without an apparent destabilization of these lipoprotein particles. Proteolysis of Lp(a) by PMN was due to an elastase activity that was 3.5% of that observed when PMN were activated by N-formyl-Met-Leu-Phe. A portion of the released elastase was found to be associated in an active form with both Lp(a) and low density lipoprotein even in an ultracentrifugal field at high salt concentrations. Taken together, our results indicate that apo(a) undergoes important proteolytic modifications by PMN elastase, which exhibits specificity for peptide bonds located in the interkringle domains of apo(a). In the case of Lp(a), elastase cleavage causes the formation of mini-Lp(a) particles with a protein moiety containing a truncated apo(a). Elastase-mediated proteolytic events may occur in vivo under conditions associated with either an excessive leakage of elastase from PMN and/or deficiencies of natural inhibitors of this enzyme.

Amino Acid Chloromethyl Ketones↗
PubMed1997

Lipoprotein(a): identification of subjects with a superbinding capacity for fibrinogen.

We have previously shown that the binding of lipoprotein(a) [Lp(a)] to immobilized fibrinogen involves the domain located in kringles IV-5 to IV-8, but not kringle IV-10. In extending those studies to subjects living in Chicago and in the island of Sardinia, we found that about 6% of them had an Lp(a) with Bmax values of 27.7+/-6.0 fmol, which were about 5-8-fold higher than those of controls (3.4+/-2.8 fmol) and in the range of those observed for free apo(a) derived from the Lp(a) of controls (36.6+/-2.9 fmol). This superbinding phenotype was unaffected by age, sex, type of lipid disorder and hypolipidemic agents, and also had a familial incidence. We are currently exploring the hypothesis that this fibrinogen superbinding phenotype is due to conformational changes of apolipoprotein(a) [apo(a)] resulting from the lipid content and composition of the Lp(a) particle and/or sequence anomalies in the kringle domain IV-5 to IV-8.

Age Factors↗
PubMed1997

Videothoracoscopic removal of a mediastinal teratoma.

We describe a case of a mature mediastinal teratoma resected by videothoracoscopy. This new procedure, now widely adopted for many intrathoracic conditions, has proved to be easy, safe, and advantageous also in the treatment of rare tumors, such as mature mediastinal teratomas. We recommend its use in all cases like the one described.

Female↗
PubMed1997

Role of hypercholesterolemia in accelerated transplant coronary vasculopathy: results of surgical therapy with partial ileal bypass in rabbits undergoing heterotopic heart transplantation.

BACKGROUND: We tested the hypothesis that plasma cholesterol lowering action of partial ileal bypass (PIB) is beneficial in mitigating accelerated transplantation coronary vasculopathy. METHODS: Forty-one New Zealand white rabbits were randomized to receive a normal (n = 21) or 1% cholesterol diet (n = 20). They underwent heterotopic heart transplantation with sham-PIB (n = 19) or PIB (n = 22) and immunosuppression with cyclosporine A (CyA). RESULTS: CyA increased plasma cholesterol of rabbits receiving a normal diet. This effect was mitigated by PIB (101 +/- 50 mg/dl CyA vs baseline 24 +/- 8, p < 0.001; vs 54 +/- 25 mg/dl with PIB, p < 0.05). In cholesterol-fed rabbits, PIB decreased plasma cholesterol levels (520 +/- 236 mg/dl PIB vs baseline 720 +/- 359, p < 0.05; vs 1502 +/- 253 mg/dl with sham PIB, p < 0.00001). Coronary arteries (CA) of 21 5-week survivors were evaluated by light microscopy and digital morphometry. No rejection was noted. Histologic study revealed vasculopathy in 3% of 705 native and 18% of 654 transplant CA (p < 0.05). Graft vasculopathy (GV) was present in 25% of 365 CA of sham-PIB and 10% of 289 CA of PIB rabbits (p = 0.07). In cholesterol-fed rabbits, GV was characterized by fatty proliferative lesions in 75% of 91 pathologic CA of sham and 21% of 28 pathologic CA of PIB rabbits (p < 0.05). Graft intimal hyperplasia was not correlated with cholesterol intake or PIB and was present in 18 of 119 pathologic CA. CONCLUSIONS: GV was characterized by fatty intimal proliferation, fibrous intimal hyperplasia, and a "mixed type." Fibrous intimal hyperplasia developed in native and transplanted hearts, and CyA seemed to promote this state. Hypercholesterolemia promoted fatty proliferative lesions, worsening GV. PIB significantly decreased total cholesterol and retarded fatty proliferation of CA of native and transplanted hearts but did not prevent intimal hyperplastic vasculopathy. Therapy of hypercholesterolemia is recommended to at least mitigate the fatty intimal proliferation of GV.

Animals↗
PubMed1997

Learning about the structure and biology of human lipoprotein [a] through dissection by enzymes of the elastase family: facts and speculations.

Lipoprotein[a], Lp[a], represents a class of lipoprotein particles that have as a protein moiety apoB-100 linked by a disulfide bridge to a multi-kringle structure, apolipoprotein[a], or apo[a]. It is now possible to separate from Lp[a] a free apo[a] able to reassociate with apoB-100-containing lipoproteins to restore the parent lipoprotein complex. Apo[a], whether free or a constitutive component of Lp[a], can be cleaved at interkringle sites by the action of enzymes of the elastase family generating fragments that differ in structural, functional, and metabolic properties. In the case of Lp[a], elastase digestion generates a miniLp[a] particle, which contains the apo[a] COOH-terminal domain able to bind to lysine, fibrinogen, fibronectin, and proteoglycans. This domain may also be generated by elastase type enzymes secreted by activated macrophages and smooth muscle cells in the arterial intima as a part of the chronic inflammation that characterizes the atherosclerotic process. Thus, the apo[a] immunoreactive material, which has been described in the atherosclerotic plaque, may represent miniLp[a] and/or apo[a] fragments accumulating in the vessel wall as a function of their relative affinity for the components of the extracellular matrix and producing complexes with an atherothrombogenic potential. This potential may depend on several factors: kringle folding and conformation, susceptibility of the linkers to proteolytic cleavage, binding specificity of given apo[a] fragments to the matrix components of the arterial intima, and the overall inflammatory status of the arterial wall.

Amino Acid Sequence↗
PubMed1996

Evidence that the fibrinogen binding domain of Apo(a) is outside the lysine binding site of kringle IV-10: a study involving naturally occurring lysine binding defective lipoprotein(a) phenotypes.

It is now established that the lysine binding site (LBS) of apo(a) kringle IV-10, and particularly Trp72, plays a dominant role in the binding of lipoprotein(a) [Lp(a)] to lysine. To determine the role of the LBS in the binding of Lp(a) to fibrinogen, we examined the binding to plasmin-modified (PM) fibrinogen of human and rhesus monkey Lp(a) species classified as either Lys' or Lys- based on their capacity to bind lysine Sepharose and to have Trp or Arg, respectively, in position 72 of the LBS of kringle IV-10. We also examined the free apo(a)s obtained by subjecting their corresponding parent Lp(a)s to a mild reductive procedure developed in our laboratory. Our results show that both Lyst and Lys- Lp(a)s and their derived apo(a)s, bound to PM-fibrinogen with similar affinities (Kds: 33-100 nM), whereas the B(max) values were threefold higher for apo(a)s. Both the lysine analog epsilon-aminocaproic acid and L-proline inhibited the binding of Lp(a) and apo(a) to PM fibrinogen. We conclude that the LBS of kringle IV-10 is not involved in this process and that apo(a) binds to PM-fibrinogen via a lysine-proline-sensitive domain located outside the LBS and largely masked by the interaction of apo(a) with apoB100. The significant difference in the PM fibrinogen binding capacity also suggests that apo(a) may have a comparatively higher athero-thrombogenic potential than parent Lp(a).

Adult↗
PubMed1996

Lys and fibrinogen binding of wild-type (Trp72) and mutant (Arg72) human apo(a) kringle IV-10 expressed in E coli and CHO cells.

In a previous study, we identified a lysine (Lys)-binding-defective form of human lipoprotein(a) and attributed this defect to the presence of a Trp72-->Arg mutation in apolipoprotein(a) [apo(a)] kringle IV-10. To document this relationship, we expressed both wild-type (wt) and mutant (mut) forms of kringle IV-10 in Escherichia coli (nonglycosylated form) and Chinese hamster ovary (CHO) cells (glycosylated form). The Arg72 mut was prepared by introducing the T-->A mutation in apo(a) kringle IV-10 amplified from human liver mRNA by the reverse-transcriptase polymerase chain reaction technique. All expressed kringles were tested for their ability to bind Lys and plasmin-modified fibrinogen (PM-fibrinogen). wt kringle IV-10 expressed in both E coli and CHO cells bound to Lys-Sepharose with comparable affinity. In contrast, the Arg72 mut expressed in both systems exhibited no Lys-binding capacity. Moreover, the wt kringle IV-10 expressed in both systems bound to PM-fibrinogen and exhibited two binding components, one Lys mediated (inhibitable by epsilon-amino-n-caproic acid) and one Lys insensitive, occurring in about the same proportions. Only the latter type of binding was present in the Arg72 mut expressed in E coli. We conclude that kringle IV-10 of human apo(a) has Lys- and PM-fibrinogen-binding capacities that are independent of glycosylation and require the presence of Trp72, one of the seven amino acids that constitute the Lys-binding site of kringle IV-10. Our results also show that the binding of kringle IV-10 to PM- fibrinogen is more complex than that to Lys, in that the former requires an additional binding site or sites outside the Lys-binding site.

Amino Acid Sequence↗
PubMed1996

Functional and metabolic differences between elastase-generated fragments of human lipoprotein[a] and apolipoprotein[a].

We have previously shown that a functional free apolipoprotein[a] (apo[a]) can be isolated from its parent lipoprotein[a] (Lp[a]) by a mild reductive procedure. To shed further light on the properties of Lp[a] and apo[a] we subjected them to a limited proteolysis by porcine pancreatic elastase. This enzyme cleaved both at the Ile3520-Leu3521 bond in the linker between kringles IV-4 and IV-5 of apo[a] generating two fragments F1 and F2. In contrast to F1, which represented the N-terminal portion of apo[a] and was functionally inert, F2, representing the C-terminal domain, bound to lysine-Sepharose, fibrinogen, and fibronectin and formed a miniLp[a] particle when incubated with LDL. The proteolytic pattern by pancreatic elastase was also exhibited by human leukocyte elastase. F1, injected intravenously into normal mice, was rapidly cleared (Ty2, 2.9 h) and after 1 h fragments in the size range of 100-33 kDa were observed in the urine. In turn, F2 had a longer residence time (Ty2, 5 h) and was excreted in the urine only after 5 h as fragments of 70-45 kDa. Fragments in the same size range as found after F1 injection were also present in the urine after injection of apo[a] or Lp[a]. Moreover, apo[a] fragments of the size seen in mouse urine were spontaneously present in normal human urine and appeared derived from larger apo[a] fragments in the plasma. Our results indicate that enzymes of the elastase family cleave human apo[a] in vitro into two main fragments that differ in structural and functional properties and metabolic behavior. The comparable size of apo[a] fragments observed in the urine of humans and injected mice invites the speculation that enzymes of the elastase family may play a role in the biology of Lp[a] in vivo.

Amino Acid Sequence↗
PubMed1995

Determinants of lipoprotein(a) assembly: a study of wild-type and mutant apolipoprotein(a) phenotypes isolated from human and rhesus monkey lipoprotein(a) under mild reductive conditions.

We previously observed that rhesus monkey lipoprotein(a) [Lp(a)], is lysine-binding defective (Lys-) and attributed this deficiency to the presence of Arg72 in the lysine-binding site (LBS) of kringle IV-10 of apolipoprotein(a) [apo(a)] [Scanu, A.M., Miles, L.A., Fless, G.M., Pfaffinger, D., Eisenbart, J., Jackson, E., Hoover-Plow, J.L., Brunck, T., & Plow, E.F. (1993) J. Clin. Invest. 91, 283-291]. We also identified human mutants having Arg72 instead of Trp72 (wild type) in the LBS of kringle IV-10 [Scanu, A M., Pfaffinger, D., lEE, J.C., & Hinman, J. (1994) Biochim. Biophys. Acta 1227, 41-45]. Unique to the human mutant phenotype were the very low levels of plasma Lp(a), suggesting structural differences between human and rhesus apo(a) and a possible divergent mode of Lp(a) assembly. In order to explore the possibility of a relationship between apo(a) LBS and Lp(a) assembly, we developed a novel method for isolating wild-type and mutant apo(a) phenotypes in a free form by subjecting each parent Lp(a) to mild reductive conditions using 2 mM dithioerythritol (DTE) and 100 mM of the lysine analogue, epsilon-aminocaproic acid (EACA). The application of this method to the study of wild-type and mutant apo(a) species showed that regardless of the source of Lp(a), i.e., positive lysine binding (Lys+) or negative lysine binding (Lys-), all of the isolated free apo(a)s were Lys+. Moreover, incubation of free apo(a)s with their autologous human or rhesus low-density lipoproteins (LDL) generated Lp(a) complexes which were structurally and functionally indistinguishable from their parent native Lp(a). In each instance, the reassembly process was inhibited by the presence of either EACA or proline. These two reagents had a minimal effect on either Lp(a) or reassembled Lp(a) [RLp(a)]. Free apo(a) bound to apoB100 of very low density lipoproteins (VLDL) to form a triglyceride-rich Lp(a). These results show that (1) both human and rhesus Lp(a) are amenable to dissassembly and reassembly, (2) the presence of Arg72 in the LBS of kringle IV-10 is not involved, at least directly, in this process, (3) its cleavage from apoB100 opens up in apo(a) a domain that is both EACA and proline sensitive and involved in Lp(a) assembly, and (4) the apoB100 of VLDL is also competent to bind apo(a). Our observations also suggest that the difference in plasma Lp(a) levels between the rhesus and the human mutant, both having Arg72 in the LBS of apo(a) kringle IV-10, is not related to the assembly process, but more likely to a divergence in production/secretion rates between the two apo(a) phenotypes.

Aminocaproates↗
PubMed1995

Identification of mutations in human apolipoprotein(a) kringle 4-37 from the study of the DNA of peripheral blood lymphocytes: relevance to the role of lipoprotein(a) in atherothrombosis.

Using a technique that amplifies the DNA region coding for kringle 4-37 of human apolipoprotein(a) we have identified 2 mutations, trp72-->arg and met66-->thr. The former was only present in 2 of the 100 subjects studied, was associated with a lysine-binding defective lipoprotein(a) [Lp(a)], low plasma levels of Lp(a), and no evidence of atherosclerotic cardiovascular disease (ASCVD). The other mutation was present in about 40% of the subjects who had either normal or high plasma levels of Lp(a) and a personal and/or familial history of ASCVD. These studies show that human kringle 4-37 is mutable and that mutations in this kringle can affect the lysine-binding properties of apo(a) and, perhaps, the atherothrombogenic potential of Lp(a).

Apolipoproteins A↗
PubMed1995

Labial mucosa and combined labial/bladder mucosa free graft for urethral reconstruction.

One-stage urethral reconstruction was performed using a free graft of labial mucosa and combined labial/bladder mucosa. We present the results of this technique in 12 cases that had a minimum follow-up period of 3 years. Eleven patients had medium penile or posterior hypospadias and one had chordee penis without hypospadias. Urethroplasty with labial mucosa was performed by two techniques: labial mucosa used alone or combined with bladder mucosa. The labial mucosa was harvested from the inner surface of the upper and/or lower lip, depending on which method was used. Seven patients, six with medium penile or posterior hypospadias and 1 with chordee without hypospadias, were given a labial mucosa graft alone; the urethral gap was 3.5 to 6 cm. The other five cases, all with posterior hypospadias, were treated by combined labial/bladder mucosa graft urethroplasty: the urethral gap was 6 to 13 cm. Follow-up (at 3 to 4 1/2 years) showed no complications apart from a urethral fistula in one patient and mild stenosis on the anastomosis in four cases, which required urethral dilatations in the first month after surgery.

Adolescent↗
PubMed1995

Lipoprotein disorders as related to atherosclerotic cardiovascular disease.

Dyslipidemias to a varying degree represent an important risk factor for atherosclerotic cardiovascular disease. They are correctable if correctly diagnosed and adequately cared for. Success depends on the optimal interplay between physician and patient and on the support of a trained dietitian. This is an area of preventive cardiology that is likely to receive increasing attention in the future, with the expectation that a decrease in morbidity/mortality for atherosclerotic cardiovascular disease will ensue.

Arteriosclerosis↗