Carotid intima-media thickness after pediatric renal or liver transplantation at high-resolution B-mode ultrasonography.

In a previous study, we observed a higher incidence of dyslipidemia in pediatric renal recipients compared with liver recipients. In the present study, we measured common carotid artery intima-media thickness (IMT) in 13 pediatric renal recipients, 9 liver recipients, and 26 control individuals with median age of 11.4, 10.8, and 12.0 years, respectively. The patients were studied from 0.2 to 10.8 years after renal transplantation (RTx) or liver transplantation (LTx). An experienced radiologist (T.K.) blinded to the status of the children measured the IMT using a high-resolution B-mode ultrasonography method. In patients who underwent RTx or LTx, serum fasting lipid profile, estimates of renal and liver function, and glucose metabolism were determined. Children undergoing RTx or LTx more often had hypertension compared with the control individuals (P = .004). Before transplantation, dyslipidemia was greater in patients undergoing RTx compared with those undergoing LTx (P < .05). Children who underwent RTx, compared with those who underwent LTx or control individuals, had thicker mean IMT at the 6 sites measured (mean [SD], 0.57 [0.07], 0.51 [0.05], and 0.53 [0.06] mm, respectively; P = .02]. As a result of linear regression in renal recipients, variability of glomerular filtration rate (<60 mL/min/1.73 m(2) vs normal) accounted for 43.3% of variability of the mean of maximal IMT (B = 8.9; SE = 3.1; P = .01). Variability of pre-RTx serum triglyceride concentration (B = 1.6; SE = 0.6; P = .03) and actual triglyceride concentration (B = 10.3; SE = 2.2; P = .002) accounted for 82.2% of variability of maximal IMT. Our findings support previous data on the importance of maintenance of good graft function with sufficient but not overly efficient immunosuppression after transplantation in prevention of future cardiovascular disease.

Male suicide mortality in eastern Finland--urban-rural changes during a 10-year period between 1988 and 1997.

AIMS: To investigate urban-rural differences in male suicide mortality between 1988 and 1997 in the province of Kuopio in eastern Finland. METHODS: Male suicide mortality between 1988 and 1997 was studied in eastern Finland. The data were collected from official autopsy reports. Age, marital status, household type, place of death, method of suicide and the region within the county were analysed. RESULTS: The age-adjusted male suicide mortality remained quite constant at 67 and 65/100,000 (> 15 years of age) in 1988 and 1997, respectively. In urban areas, a decline in suicide rates was noted in all age groups of men, whereas in rural areas, after an initial decline, a marked increase took place during the last years of the study period, especially among middle-aged and elderly men. This coincided with the time of recovery from an economic recession in Finland that particularly affected rural areas. CONCLUSION: Male suicide mortality may be regionally diverging in Finland.

Skeletal muscle protein mass correlates with the lipid status in children with solid tumors and before bone marrow transplantation.

OBJECTIVE: To evaluate the changes in lipid status in children during anticancer therapy, with special reference to the effect of protein-energy malnutrition on plasma lipids. DESIGN: Prospective follow-up study. SETTING: The study was carried out in the Hospital for Children and Adolescents, University of Helsinki, Helsinki, Finland SUBJECTS: The study group consisted of 33 children going through bone marrow transplantation (BMT) and 10 children with malignant solid tumors. The BMT patients were evaluated before transplantation and 1 and 3 months after BMT, and the tumor patients were studied at diagnosis and in remission. The reference group consisted of 23 healthy children. INTERVENTIONS: As indicators of lipid status, lipoproteins and the concentration of cholesterol and triacylglycerol were measured. Protein reserves were expressed as muscle index (MI), derived from ultrasonographic measurement of the femoral quadriceps muscle. Body weight, triceps skinfold thickness and the serum concentration of albumin, prealbumin and transferrin were measured. RESULTS: In both groups, plasma concentration of total triacylglycerol was increased and high-density lipoprotein (HDL) cholesterol decreased as compared to the reference subjects. Plasma triacylglycerol concentration had a negative correlation with skeletal muscle protein mass (MI; r=0.34, P=0.02). The concentration of serum prealbumin correlated positively with plasma total cholesterol concentration (r=0.47, P=0.002). CONCLUSIONS: In children with cancer, abnormalities of lipid status are associated with changes in skeletal muscle protein reserves. SPONSORSHIP: This study was supported by the Foundation of Pediatric Research, Helsinki, Finland and the Nona and Kullervo Väre Foundation, Helsinki, Finland.

Association of variation in hepatic lipase activity with promoter variation in the hepatic lipase gene. The LOCAT Study Invsestigators.

The associations between six genetic polymorphisms in the hepatic lipase (HL) gene (LIPC) and variation in postheparin HL activity and fasting serum lipoproteins were evaluated in 395 male Finnish coronary heart disease patients with HDL cholesterol concentrations

The Asn-291-->Ser and Ser-477-->Stop mutations of the lipoprotein lipase gene and their significance for lipid metabolism in patients with hypertriglyceridaemia.

We examined 99 Finnish patients whose serum fasting triglycerides (TG) had exceeded 6.0 mmol L-1 with special interest to their lipid, lipoprotein and post-heparin plasma lipase activities. The control group consisted of 75 healthy individuals. We also determined the frequency of the Asn-291-->Ser and Ser-447-->Stop mutations both in hypertriglyceridaemic (HTG) subjects and in control subjects. A total of 51 of the original 99 hypertriglyceridaemic patients still had TG > 6.0 mmol L-1 when measured a second time. They are referred to as persistently hypertriglyceridaemic subjects (pHTG). The remaining 48 subjects had TG < 6.0 mmol L-1 in the second measurement and are referred to as sporadically hypertriglyceridaemic subjects (sHTG). The allelic frequencies of the Ser-447-->Stop mutation in the total HTG and sHTG groups were similar to the frequencies present in the control group, but lower in pHTG patients compared with the control group (0.049 vs. 0.153, chi(2) = 6.63, P < 0.05). The Asn-291-->Ser mutation was more frequent in HTG group than in the control group (0.0606 vs. 0.013, chi(2) = 4.86, P < 0.05). This difference was due to the higher frequency of the minor allele of Asn-291-->Ser in the cohort with persistent hypertriglyceridaemia compared with the control group (0.088 vs. 0.013, chi(2) = 8.00, P < 0.01). The highest frequency (0.114) of the minor allele of Asn-291-->Ser was found in type 2 diabetic patients with persistent hypertriglyceridaemia. The carrier status of Asn-291-->Ser or Ser-447-->Stop did not predict either post-heparin plasma lipoprotein lipase (LPL) activities or lipid and lipoprotein levels in any of the groups studied. Our data suggest that overproduction of very low-density lipoproteins (VLDL) is a more important cause of hypertriglyceridaemia in the Finns than is the LPL deficiency.

Hepatic lipase gene polymorphisms influence plasma HDL levels. Results from Finnish EARS participants. European Atherosclerosis Research Study.

Hepatic lipase (HL), a triglyceride lipase found in liver, adrenals, testes, and ovaries, takes part in the uptake, remodeling, and function of lipoproteins including HDL, as well as VLDL and chylomicrons. In the present study, the genotype distribution of five HL polymorphisms (-C480T, V133V, T202T, L334F, T457T) and their association to plasma lipid values were investigated. The study participants included 92 students with paternal history of myocardial infarction before the age of 55 and 194 matched control subjects, ie, the Finnish participants of the European Atherosclerosis Research Study (EARS). The allele T of the HL polymorphism -C480T showed an association with elevated HDL, apoA-I, and LpA-I values (ANOVA P < .01). No difference in genotype distribution was observed in the offspring with and without paternal history of myocardial infarction.

Phenotype expression in familial combined hyperlipidemia.

Familial combined hyperlipidaemia (FCHL) is one of the most common hereditary disorders predisposing to early coronary death. The affected family members have elevations of serum total cholesterol, triglycerides or both. Despite intensive research efforts the genetic and metabolic defects underlying this complex disorder are still unknown. To dissect the metabolism and genetics of FCHL the phenotype of an individual must be precisely defined. We assessed the influence of different diagnostic criteria on the phenotype definition and studied factors affecting the phenotype expression in 16 large Finnish families (n = 255) with FCHL. The fractile cut-points used to define abnormal lipid values had a profound influence on the diagnosis of FCHL. If the 90th percentile cut-point was used, approximately 45% of the family members were affected, in concord with the presumed dominant mode of transmission for FCHL. If the 95th percentile was used only 22% of study subjects were affected. To characterize the metabolic differences or similarities between the different lipid phenotypes, we determined very low density lipoprotein (VLDL), intermediate density lipoprotein (IDL), low density lipoprotein (LDL) and high density lipoprotein (HDL) particles separated by ultracentrifugation. In linkage analysis no single ultracentrifugation variable could discriminate reliably affected family members from non-affected family members. Our data emphasizes the need for re-evaluation of FCHL diagnostic criteria. Preferably, the diagnosis should be based on a single, reliable metabolic marker.

No evidence of linkage between familial combined hyperlipidemia and genes encoding lipolytic enzymes in Finnish families.

Familial combined hyperlipidemia (FCHL) is characterized by different lipid phenotypes (IIa, IIb, IV) and elevated apolipoprotein B (apo B) levels in affected family members. Despite intensive research, the genes involved in the expression of this complex disorder have not been identified, probably because of problems associated with phenotype definition, unknown mode of inheritance, and most probably genetic heterogeneity. To explore the genetics of FCHL in the genetically homogeneous Finnish population, we collected 14 well-documented Finnish pedigrees with premature coronary heart disease and FCHL-like dyslipidemia. The lipolytic enzymes lipoprotein lipase (LPL), hepatic lipase (HL), and hormone-sensitive lipase (HSL) were selected as initial candidate genes because of their central roles in apo B and triglyceride metabolism. On the basis of the pedigree structures, a dominant mode of inheritance was adopted for linkage analyses, and serum total cholesterol and/or triglyceride levels exceeding the 90th percentile level were set as diagnostic criteria (criterion 1). In pairwise linkage analyses with intragenic markers, no evidence for linkage was found. Instead, the significantly negative LOD scores suggested exclusion of all three loci for single major gene effect. LOD scores were -14.63, -5.03, and -5.70 for the three LPL polymorphisms (theta=0.00); -9.40, -6.30, and -4.74 for the three HL polymorphisms (theta=0.00); and -15.29 for the HSL polymorphism (theta=0.00). The results were very similar when apo B levels over the 90th percentile were used as criteria for affected status (criterion 2). Also, when linkage calculations were carried out using an intermediate or recessive mode of inheritance, the results of pairwise linkage analysis remained negative. Furthermore, when haplotypes were constructed from multiple polymorphisms of the LPL and HL genes, no segregation with the FCHL phenotype could be observed in the 14 Finnish families. Data obtained by the affected sib-pair method supported these findings, suggesting that the LPL, HL, or HSL genes do not represent major loci influencing the expression of the FCHL phenotype.

Heterozygous hepatic lipase deficiency, due to two missense mutations R186H and L334F, in the HL gene.

Hepatic lipase (HL) is an endothelial enzyme involved in the metabolism of intermediate density lipoproteins (IDL) and high density lipoproteins (HDL) in plasma. In a Finnish pedigree consisting of 18 members belonging to three generations two missense mutations RI86H and L334F in exons 5 and 7 of the HL gene co-segregated with low post-heparin HL activity. Haplotype analysis of the HL gene in family members revealed a high degree of genetic variation and demonstrated that the two missense mutations reside on the same chromosome. In vitro site-directed mutagenesis and expression of the cDNA constructs in COS-1 cells revealed that the R186H mutation leads to a protein that is not secreted while the L334F mutation results in the production of a HL protein that is secreted but has only about 30% of wild type HL activity. Carriers of the mutated HL gene exhibited clearly reduced HL activity and mass in post-heparin plasma. Probably due to their heterozygous carrier status they had only moderate elevation of total triglycerides, IDL, and LDL-triglycerides. The LDL-particles were enriched in triglycerides and depleted of cholesterol. Also their HDL2- and HDL3-particles were enriched in triglycerides.

Peritoneal dialysis in children under 5 years of age.

OBJECTIVE: We report our experience with maintenance peritoneal dialysis (PD) in small children. DESIGN: This is a retrospective analysis of the patient records of all children under the age of 5 years treated with continuous peritoneal dialysis (CPD) between 1986 and 1994 in Finland. SETTING: Treatment was started and the patients were seen at the outpatient clinic at the Hospital for Children and Adolescents, University of Helsinki, every 3 months. Between these visits, they had controls at their local hospital every 2-4 weeks. PATIENTS: The most common primary renal disease in these 34 patients was congenital nephrotic syndrome of the Finnish type (27 patients). Others were: congenital nephrotic syndrome (3 patients), polycystic kidney disease (1), urethral valve (1), neuroblastoma (1), and renal dysplasia (1). RESULTS: Mean age at onset was 1.6 years and median treatment time 9.3 months. Time spent in hospital decreased from 270 days/year in the 1980s to 150 days/year in the 1990s. Two children died (5.9%). The peritonitis rate on continuous cyclic peritoneal dialysis was 1:11.5 patient-months. Hernias were diagnosed in 29% of the patients. After 3 months half of the patients were on antihypertensive medication. Pulmonary edema was diagnosed once in 12 patients and twice in 2 patients. During the first 6 months on PD the mean height standard deviation score (hSDS) increased from -2.13 to -1.66 (p < 0.0001). The 6-month change in hSDS before initiation and 6 months after the start of CPD increased from -0.12 +/-0.68 to +0.59 +/- 0.64 (p = 0.0008). CONCLUSIONS: Our results indicate that peritoneal dialysis is feasible and safe in small children. Mortality was low and growth was good. The major challenges presented by CPD therapy were maintenance of optimal nutrition, avoidance of peritonitis, and control of volemia.

The Gln-Arg191 polymorphism of the human paraoxonase gene (HUMPONA) is not associated with the risk of coronary artery disease in Finns.

The human paraoxonase gene (HUMPONA) is codominantly expressed as alleles A and G. The A allele codes for glutamine (A genotype) and the G allele for arginine (B genotype) at position 191 of the paraoxonase enzyme. This genetic polymorphism has been suggested to be associated with the predisposition to coronary artery disease (CAD). We investigated the frequency of paraoxonase A and G alleles in 380 well-characterized CAD patients and in 169 controls. The most common genotype in both the patients with CAD (211/380) and in healthy Finnish individuals (87/169) was AA (Gln/Gln). The heterozygous AM (Gln/Arg) genotype was present in 140 of the patients and in 75 controls. The frequency of the A allele was 0.74 in both patients and controls. The genotype distribution between the two groups did not differ (P = 0.12, chi2 test). The genotype distributions were also similar to those reported earlier in other caucasoid populations. In conclusion, we found no association between the Gln-Arg 191 polymorphism of the human paraoxonase gene and coronary artery disease in Finns.

Development and evaluation of an ELISA method for the determination of lipoprotein lipase mass concentration--comparison with a commercial, one-step enzyme immunoassay.

We developed a non-competitive, enzyme-linked, immunosorbent assay (ELISA) for the quantitation of lipoprotein lipase (LPL) in human postheparin plasma using affinity-purified antihuman milk lipoprotein lipase antibodies produced in chicken eggs and a monoclonal antibody directed against human lipoprotein lipase. We compared our ELISA method with a commercially available sandwich-enzyme immunoassay (Markit-F LPL EIA Kit, Dainippon Pharmaceutical Co, Ltd. Osaka, Japan). The reference values for lipoprotein lipase catalytic activity concentration and mass concentration in healthy Finns were determined. Lipoprotein lipase activity concentration (mean +/- SD) was 297 +/- 112 U/l in women, and mass concentration as measured by the ELISA method was 1058 +/- 367 micrograms/l. The corresponding values for men were 247 +/- 97 U/l and 815 +/- 207 micrograms/l, respectively. Across the whole concentration range of the ELISA method, the control samples' intra- and inter-assay coefficients of variation (CV) were 5.1% and 6.5%, respectively. The correlation between the ELISA and EIA methods was good, r = +0.81. The importance of the correct standardisation of immunoassays is discussed.

Heterozygosity for Asn291-->Ser mutation in the lipoprotein lipase gene in two Finnish pedigrees: effect of hyperinsulinemia on the expression of hypertriglyceridemia.

We describe two Finnish kindreds with the Asn291 --> Ser mutation (A291S) of the lipoprotein lipase (LPL) gene. Sixteen subjects (9 male, 7 female) heterozygous for this mutation were studied and compared with 17 unaffected family members or spouses (family controls) and 19 unrelated healthy subjects (population controls). In the group of subjects heterozygous for the A291S mutation, postheparin plasma LPL activity was on average 23% lower than in the family controls and 29% lower than in the population controls. In agreement, in vitro expression studies with COS-7 cells showed that the mutant protein exhibits approximately 50% of the lipolytic activity of the wild-type protein. Median serum triglyceride concentration was 2.90 mmol/l in the group of heterozygotes, compared with 1.14 mmol/l in the family controls (P < 0.01) and 0.99 mmol/l in the population controls (P < 0.001). The heterozygotes also had a marked preponderance of small dense low density lipoproteins (LDL) as assessed by gradient gel electrophoresis. Nine of the heterozygous subjects were hypertriglyceridemic (serum triglyceride concentration > 2.0 mmol/l). Age or body mass index were not related to the presence of hypertriglyceridemia. By contrast, all hypertriglyceridemic subjects were either hyperinsulinemic (serum insulin concentration > 10 mU/l, n = 7) or had diabetes (n = 2). In a multivariate regression analysis, very low density lipoprotein (VLDL) triglyceride level was significantly and independently related to serum apolipoprotein B concentration, the presence of the A291S mutation, serum insulin concentration, and postheparin plasma LPL activity. The Asn291-->Ser mutation of the LPL gene results in reduced lipolytic activity. However, dyslipidemia appears to manifest only if VLDL production is also increased. Hyperinsulinemia was the major determinant of excessive VLDL synthesis and dyslipidemia among the subjects heterozygous for the A291S mutation in this study.

A compound heterozygote for hepatic lipase gene mutations Leu334-->Phe and Thr383-->Met: correlation between hepatic lipase activity and phenotypic expression.

We have characterized the molecular basis for familial hepatic lipase (HL) deficiency in a Finnish family. In the propositus, the HL deficiency results from compound heterozygosity for two rare HL gene mutations, a previously unknown missense mutation designated L334F and the previously reported T383M mutation. These mutations were introduced into human HL cDNA by site-directed mutagenesis and the constructs expressed in COS-1 cells. In the homogenate of COS-1 cell transfected with the L334F mutant cDNA, a high amount of inactive protein accumulated. In the media of L334F transfected cells, 30% of the wild type activity and 80% of wild type mass were detected. The lysates of COS-1 cells transfected with the T383M mutant cDNA contained 39% of wild type HL activity and 34% of wild type HL mass. In the media of COS-1 cells transfected with the T383M cDNA construct, 50% of wild type HL mass but only 6% of wild type activity was present. The single amino acid substitutions present in L334F and T383M are therefore sufficient to severely affect the HL enzyme. These defects explain the HL-deficient phenotype of the individual carrying the two mutations. The lipoprotein phenotype associated with compound heterozygosity for L334F and T383M mutations is characterized by a slight increase in the buoyant low density lipoprotein (LDL) fraction and an increase in the light high density lipoprotein (HDL) fractions, HDL2a and HDL2b. These results demonstrate that lipoprotein changes occurring in HL deficiency are difficult to identify and support the hypothesis that HL is important in HDL remodeling and metabolism in vivo.

Immunolocalization of Antifreeze Proteins in Winter Rye Leaves, Crowns, and Roots by Tissue Printing.

During cold acclimation, antifreeze proteins (AFPs) that are similar to pathogenesis-related proteins accumulate in the apoplast of winter rye (Secale cereale L. cv Musketeer) leaves. AFPs have the ability to modify the growth of ice. To elucidate the role of AFPs in the freezing process, they were assayed and immunolocalized in winter rye leaves, crowns, and roots. Each of the total soluble protein extracts from cold-acclimated rye leaves, crowns, and roots exhibited antifreeze activity, whereas no antifreeze activity was observed in extracts from nonacclimated rye plants. Antibodies raised against three apoplastic rye AFPs, corresponding to a glucanase-like protein (GLP, 32 kD), a chitinase-like protein (CLP, 35 kD), and a thaumatin-like protein (TLP, 25 kD), were used in tissue printing to show that the AFPs are localized in the epidermis and in cells surrounding intercellular spaces in cold-acclimated plants. Although GLPs, CLPs, and TLPs were present in nonacclimated plants, they were found in different locations and did not exhibit antifreeze activity, which suggests that different isoforms of pathogenesis-related proteins are produced at low temperature. The location of rye AFPs may prevent secondary nucleation of cells by epiphytic ice or by ice propagating through the xylem. The distributions of pathogenesis-induced and cold-accumulated GLPs, CLPs, and TLPs are similar and may reflect the common pathways by which both pathogens and ice enter and propagate through plant tissues.

Management of congenital nephrotic syndrome of the Finnish type.

Congenital nephrotic syndrome of the Finnish type (CNF) is a rare autosomal recessively inherited disease characterised by intrauterine onset of massive urinary loss of proteins, 90% of which is albumin. The CNF gene has been localised to the long arm of chromosome 19, but the pathogenesis remains unclear. Historically, all CNF patients died, usually within the first 6 months of life. Today, a normal life can be achieved for a child with CNF by correcting the protein deficiency and normalising nutrition. This is accomplished by early intravenous albumin supplementation, nutritional support, aggressive treatment of complications and early renal transplantation, after bilateral nephrectomy and peritoneal dialysis. In the present article current treatment strategies are reviewed, and our own experience with 43 CNF patients during the last 10 years is presented.

Short-term effects of renal transplantation on plasma lipids and lipoprotein lipase in children with congenital nephrosis.

Plasma lipoprotein and apoprotein (apo) A-I, A-II and B concentrations and post-heparin plasma lipoprotein lipase (LPL) activity and mass were measured in 13 children with congenital nephrotic syndrome of the Finnish type (CNF) six months after renal transplantation to determine whether the severe lipid abnormalities documented prior to and during peritoneal dialysis would normalize. Plasma total triglyceride decreased by 52% (p < 0.001), VLDL triglyceride by 55% (p < 0.01) and total cholesterol by 17% (p < 0.01). HDL cholesterol increased by 51% (p < 0.001), whereas no significant change was observed in LDL cholesterol. Despite these improvements, plasma lipoprotein concentrations were still abnormal after transplantation. Total (p < 0.01) and VLDL triglyceride (p < 0.05) as well as total (p < 0.01), VLDL (p < 0.01) and LDL cholesterol (p < 0.01) were higher than in control children. HDL cholesterol normalized. Plasma apo A-I and A-II concentrations were normal, but the apo B concentration remained high (p < 0.01). Post-heparin LPL activity and mass were normal (25.0 +/- 9.1 mumol FFA/ml/h and 227.2 +/- 88.4 ng/ml). The mean cyclosporine dose correlated positively with the serum creatinine concentration (r = +0.72, p < 0.01). Positive correlations were also observed between total (r = +0.68, p < 0.05) and VLDL triglyceride (r = +0.62, p < 0.05) and the serum creatinine concentrations. We conclude that renal transplantation substantially improves the triglyceride and cholesterol abnormalities in CNF but significant abnormalities still persist.

A novel amino acid substitution (His183-->Gln) in exon 5 of the lipoprotein lipase gene results in loss of catalytic activity: phenotypic expression of the mutant gene in a heterozygous state.

We have identified a hitherto unrecognized mutation of the lipoprotein lipase gene (LPL) in a Finnish family with Russian and Swiss ancestors. A single base pair substitution of a guanine for cytosine in codon 183 of exon 5 of the LPL gene results in a change of histidine to glutamine in the mature enzyme protein. Expression of a mutant cDNA construct in COS cells resulted in secretion of inactive LPL enzyme protein confirming the functional significance of the mutation. The proband, a 50-year-old female and her two daughters were all heterozygous for the His183-->Gln mutation. Clinically, the proband was characterized by variable and occasionally severe hypertriglyceridemia, obesity, hypertension, coronary heart disease and non-insulin-dependent diabetes mellitus. The daughters, aged 24 and 19 years, were also obese but had milder hypertriglyceridemia. In conclusion, we have identified a novel LPL mutation that results in the synthesis of an inactive enzyme protein. Although the assessment of a causative link between the mutation and hyperlipidemia awaits further studies, our data suggest that heterozygosity for a functional defect of LPL should be considered in patients presenting with the metabolic dyslipidemic syndrome, "syndrome-X."