Effect of vitamin D receptor gene (VDR) polymorphism on body height in children - own experience.

UNLABELLED: Genetic and environmental factors have an influence on the process of growth and development of the body. One of numerous genetic factors can be the vitamin D receptor gene (VDR). The study aimed at evaluating the relationship between VDR polymorphism and somatic parameters in children. PATIENTS AND METHODS: The study group consisted of 395 children, aged 6-18 years. All the patients underwent gene typing using the PCR-RFLP method within polymorphic loci BsmI (rs1544410), FokI (rs2228570), ApaI (rs7975232) and TaqI (rs731236) of the VDR receptor gene. 294 children made up the control group in the study on the incidence of particular genotypes; in 161 patients somatic measurements of body weight and height were made with standard methods and skeletal densitometry (total body and spine programmes) examination was performed. Statistica 10.0 PL was used for statistical analysis. RESULTS: In patients with low bone mass a relationship between body height and FokI VDR polymorphism was noted. The p-value was statistically significantly different in group I (p=0.002) and borderline significant in group III (p=0.09). None of the polymorphisms of the VDR receptor gene demonstrated any statistically significant differences in anthropometric values in the control group and in children with osteoporosis. SUMMARY: The presence of the F allele of FokI polymorphism of the VDR receptor gene results in increased height, which is best observed in children with low bone mass. The FF genotype favours increased height in the study group of children from Lódz.

The prion protein M129V polymorphism: longevity and cognitive impairment among Polish centenarians.

The PRNP gene encodes the cellular isoform of prion protein (PrP (c) ). The M129V polymorphism influences the risk of prion diseases and may modulate the rate of neurodegeneration with age. We present the first study of the polymorphism among Polish centenarians. In the control group (n = 165, ages 18 to 56 years) the observed M129V genotype frequencies agreed with those expected according to the Hardy-Weinberg equilibrium (MM, MV, VV): 43%, 44%, 13% (HWE p > 0.05). Among centenarians (n = 150, ages 100 to 107) both homozygotes were more common than expected and HWE was rejected: 46%, 37%, 17% (expected 42%, 46%, 13%; HWE p = 0.025). This finding is consistent with a higher mortality rate among heterozygotes. However, the observed allele and genotype frequencies did not differ significantly between the oldest-old and the young controls. The genotypic frequencies were not related to severe cognitive impairment among the centenarians.

APBB2 genetic polymorphisms are associated with severe cognitive impairment in centenarians.

APBB2 gene encodes for ß-amyloid precursor protein-binding family B member 2, (APBB2, FE65-like, FE65L1), an adaptor protein binding to the cytoplasmatic domain of ß-amyloid precursor protein (ßAPP). Over-expression of APBB2 promotes formation of ß-amyloid (Aß), the main constituent of senile plaques. Polymorphisms within APBB2 gene have been proposed as candidate risk factors for Alzheimer's disease. However, their association with longevity has never been investigated. Here we present the first attempt to analyze APBB2 polymorphisms in centenarians. We used a PCR-RFLP method to analyze two intronic nucleotide substitutions: hCV1558625 (rs17443013) and rs13133980. We found no differences in genotype or allele distribution between centenarians and young controls. After stratification of centenarians upon their cognitive performance, the APBB2 rs13133980 G allele was over-represented in centenarians with severe cognitive impairment compared to individuals without this disability. Also the hCV1558625-rs13133980 AG haplotype increased relative risk for severe cognitive impairment in centenarians. Our results support the concept of APBB2 polymorphism association with cognitive performance in the oldest age.

Vitamin D receptor gene variability as a factor influencing bone mineral density in pediatric patients.

To determine the relationship between the polymorphism of vitamin D receptor gene and the bone mineral density in children. The study group consisted of 395 children aged 6-18 years. All patients underwent genotyping using the PCR-RFLP method within polymorphic loci BsmI (rs1544410), FokI (rs2228570), ApaI (rs7975232) and Taq I (rs731236) of the VDR gene. The BMD (g/cm(2), Z score) and BMC (g, Z score) by DXA method, as well as Z scores of the BUA, SOS and Stiffness ultrasound parameters were evaluated. Based on densitometry results, children were divided into 3 groups: I-Z score ± 1.0; II-Z score from -1.1 to -2.0; and III-Z score ≤ -2.1. A control group numbering 294 children was used for the purpose of allele frequency comparisons. The occurrence of studied polymorphism alleles in the control group did not significantly differ from the values expected according to the Hardy-Weinberg equilibrium (p values: 0.1224 for BsmI; 0.5958 for TaqI; 0.0817 for ApaI; and 0.8901 for FokI). Allele a ApaI carrier status in group III children was associated with an increased BMD (x = 0.8 vs 0.69, p = 0.0296) and BMC value (x = 28.76 vs 22.14, p = 0.0565) in spine projection results, Stiffness (x = -1.12 vs -1.91, p = 0.0347) and SOS (x = -1.43 vs -2.27, p = 0.0319) ultrasound parameters. In group II, significantly increased SOS values (-1.13 vs -1.73, p = 0.0378) were noted in f (FokI) carriers. The presence of aa ApaI and ff FokI polymorphisms favours a higher bone mass and better bone structure (decreased bone mass loss) in the analysed group.

Increased risk of type 1 diabetes in Polish children - association with INS-IGF2 5'VNTR and lack of association with HLA haplotype.

BACKGROUND: Human leukocyte antigens (HLA) complex and INS-IGF2 5'VNTR loci are principal determinants of the risk of type 1 diabetes mellitus (T1DM). Carriage of class III allele is protective, while class I/I homozygosity increases the risk of T1DM. MATERIAL AND METHODS: HLA and 5'VNTR allele frequencies were summarised and multivariate logistic regression models with interaction evaluation were employed to determine the presence and types of allele effect interdependency. The study group was planned to number 590 children who would undergo genotyping of 5'VNTR and HLA. RESULTS: 590 patients (302 with T1DM and 288 controls) were recruited. Frequencies of HLA risk alleles were: 117 carriers of DR3-DQ2; 130 carriers of DR4-DQ8 including 43 DR3-DQ2/DR4-DQ8 heterozygotes. In all cases, risk alleles were vastly overrepresented in the T1DM group compared to the controls (p < 0.0001 in all cases). The most frequent protective haplotype was DQB1 × 0602 observed in 24 controls and two T1DM cases (p < 0.001). Class I 5'VNTR homozygotes constituted 58% of the control group (n = 174) and 78% (n = 224) of T1DM patients [OR = 2.63 (95% CI: 1.79-3.57)]. Interactions between 5'VNTR and DR3-DQ2 or DR4-DQ8 variants did not reach statistical significance for risk of developing T1DM (p = 0.54 and 0.24) or age at its diagnosis (p = 0.14 and 0.67 respectively). CONCLUSIONS: Interactions between HLA and 5'VNTR genotype are not of multiplicative character. Class I homozygosity at 5'VNTR is a significant risk factor of T1DM and acts independently from HLA haplotype in determining the actual risk of diabetes in children.

Cell recovery comparison between plasma depletion/reduction- and red cell reduction-processing of umbilical cord blood.

BACKGROUND AIMS: Limited cell dose has hampered the use of cord blood transplantation (CBT) in adults. One method of minimizing nucleated cell loss in cord blood (CB) processing is to deplete or reduce plasma but not red blood cells - plasma depletion/reduction (PDR). METHODS: The nucleated cell loss of PDR was studied, and determined to be less than 0.1% in the discarded supernatant plasma fraction in validation experiments. After testing and archival sampling, the median nucleated cell recovery for PDR processing was 90%, and median CD34(+) cell recovery 88%. In a CB bank inventory of 12 339 products with both pre- and post-processing total nucleated cells (TNC), PDR processing resulted in median post-processing TNC recoveries of 90.0% after testing and archival samples removal. Using the same 10 CB units divided into two halves, we compared directly the recovery of PDR against hydroxyethyl starch red cell reduction (RCR) for TNC, CD34(+) cells and colony-forming units (CFU-GM, CFU-E, CFU-GEMM and total CFU) after parallel processing. We also compared the loss of very small embryonic-like stem cells (VSEL). RESULTS: We demonstrated significantly higher recoveries using PDR for TNC (124%), CD34(+) cells (121%), CFU-GM (225%), CFU-GEMM (201%), total CFU (186%) and VSEL (187%). The proportion of high TNC products was compared between 10 912 PDR and 38 819 RCR CB products and found to be 200% higher for products that had TNC ≥150 × 10(7) (P = 0.0001) for the PDR inventory. CONCLUSIONS: Our data indicate that PDR processing of CB provides a significantly more efficient usage of this valuable and scarce resource.

Effect of the IP10 (CXCL10) and HLA genotype on the risk of type 1 diabetes in children.

INTRODUCTION: Chemokines may modulate autoimmunity through a variety of mechanisms. Recent reports suggest involvement of the IP10 (CXCL10) chemokine in autoimmunity towards pancreatic ß cells. AIM OF THE STUDY: The purpose of this study was to evaluate the effects of genetic variability of IP10 and its dependence on HLA-conferred risk of diabetes. MATERIALS AND METHODS: 148 children with type 1 diabetes and 105 healthy adult controls were evaluated. Both groups underwent HLA and IP10 genotyping at rs8878. This polymorphic site is a tagging SNP of a haploblock encompassing the whole 3' untranslated region of the IP10 gene. RESULTS: T allele homozygosity was protective against type 1 diabetes (Odds Ratio (OR) 0.48 (95% confidence interval (95%CI) 0.23-0.99). No conditional interaction effect was observed in carriers of the T allele (T+) with high risk HLA genotypes: OR(95%CI)=0.65 (0.86-1.68) for DQ2∗T+ genotype and OR(95%CI)=1.20 (0.37-1.16) for DQ8∗T+ genotype respectively. An interaction effect promoting earlier onset of diabetes was observed in individuals with DQ8∗T+ genotypes. In these patients diabetes was diagnosed at a mean age of 5.1 (95%CI 3.3-6.8) in comparison to 9.0 (95%CI 7.6-10.4) years in patients with other genotypes (p=0.03). CONCLUSIONS: Genetic variability of the IP10 gene may play a varied role depending on the inherent genetic risk of autoimmunity. IP10 (CXCL10) variability may reduce the risk of type 1 diabetes or alter its course depending on HLA background.

ICOS gene polymorphism may be associated with pemphigus.

BACKGROUND: Pemphigus is an autoimmune blistering disease mediated by circulating IgG autoantibodies directed against desmogleins 3 and/or 1. As pemphigus is a T cell-mediated disease, one may assume that genetically determined disregulation of costimulatory signal may be involved in its pathogenesis. OBJECTIVE: The aim of the present study was to evaluate the relationship between polymorphisms in genes encoding costimulatory receptors, CTLA4 and ICOS, and pemphigus in the Polish population. METHODS: The study included 54 patients with pemphigus: 40 with pemphigus vulgaris (PV) and 14 with pemphigus foliaceus (PF). Additionally, 176 healthy unrelated blood donors served as controls. +49A/G CTLA4 and IVS1+173 ICOS polymorphisms were identified using a modified polymerase chain reaction-restriction fragment-length polymorphism. RESULTS: Analysis of the frequency of genotypes and alleles of +49A/G CTLA4 gene polymorphism showed no statistically significant differences between the PV and PF patients and the controls. The distribution of genotypes in IVS1+173 ICOS polymorphisms was significantly different in both PV (p < .01) and PF (p  =  .0004) patients when compared to controls. The carriers of the allele C were more frequent in PV or PF in comparison with the control group (p < .001 for both groups). CONCLUSIONS: Our results suggest that genetically determined abnormal function of costimulatory receptors in T cells may be associated with the pathogenesis of pemphigus.

Molecular characterization of isolated from murine adult tissues very small embryonic/epiblast like stem cells (VSELs).

Pluripotent very small embryonic/epiblast derived stem cells (VSELs) as we hypothesize are deposited at begin of gastrulation in developing tissues and play an important role as backup population of pluripotent stem cells (PSCs) for tissue committed stem cells (TCSCs). We envision that during steady state conditions these cells may be involved in tissue rejuvenation and in processes of regeneration/repair after organ injuries. Molecular analysis of adult bone marrow (BM)-derived purified VSELs revealed that they i) express pluripotent stem cells markers e.g., Oct4, Nanog, Klf-4, SSEA-1 ii) share several markers characteristic for epiblast as well as migratory primordial germ cells (PGCs), and iii) possess a unique pattern of genomic imprinting (e.g., erasure of differently methylated regions at Igf2-H19 and Rasgrf1 loci and hypermethylation at KCNQ1 and Igf2R loci). This supports that VSELs are related to epiblast-derived migrating PGC-like cells and, despite their pluripotent stem cell character, changes in the epigenetic signature of imprinted genes keep these cells quiescent in adult tissues and prevent them from teratoma formation. In contrast epigenetic changes/mutations that lead to activation of imprinted genes could potentially lead to tumor formation by these cells. Mounting evidence accumulates that perturbation of expression of imprinted genes is a common phenomenon observed in developing tumors.

[An evaluation of HLA class 2 alleles and anti-islet antibodies as evidence for non-autoimmune diabetes in Wolfram syndrome]. / Analiza alleli HLA klasy 2 i przeciwcial przeciw antygenom komórek B jako dowód na nieautoimmunologiczna patogeneze cukrzycy w zespole Wolframa.

INTRODUCTION: A clinical criterion of the Wolfram syndrome is the coexistence of diabetes and optic atrophy recognized before the age of 15. Diabetes present in Wolfram syndrome is a result of the selective ß cell loss and failed insulin secretion which is probably associated with non-autoimmune pathogenesis. AIM OF THE STUDY: The aim of the study was an evaluation of HLA subtypes and presence of ß-cell autoantibodies in patients with molecularly confirmed Wolfram syndrome. MATERIAL AND METHODS: 9 patients with Wolfram syndrome aged 10-24 years were examined. We also studied 218 patients with type 1 diabetes as a reference group. A control group of 176 healthy individuals was included in the study. Besides the clinical assessment the HLA typing by PCR-SSO was performed. Islet cell antibodies (ICA), antibodies to glutamic acid decarboxylase (GADA), thyrosine phosphatase antibodies (IA2A) and insulin antibodies (IAA) were also detected. RESULTS: In all nine patients the coexistence of diabetes with optic atrophy was observed and in 8/9 individuals additional symptoms were recognized. In patients with Wolfram syndrome a significantly lower age of diagnosis of diabetes (Me=5.0 years) than in type 1 diabetic children (Me=10.4; p=0.002) was observed. Studies of HLA subtypes demonstrated an increased prevalence of HLA-DQw1, DRB1⋅03 and/or 04 and DR2. A comparison of the frequency of the HLA alleles in patients with Wolfram syndrome with type 1 diabetic children showed a more frequent presence of the DRB1⋅1501 (p=0.03; OR=13.28 (2.44-72.12)) and DQB1⋅06 (p=0.016; OR=10.15 (2.49-41.35)) alleles in patients with Wolfram syndrome. CONCLUSIONS: Polish patients with Wolfram syndrome have a different profile of the HLA antigens with the presence of DR2, DQw1 and DRB3/4 allele and are negative for diabetes-related autoantibodies, which may confirm non-autoimmune ß-cell destruction in this syndrome.

Erythrocyte-derived microvesicles may transfer phosphatidylserine to the surface of nucleated cells and falsely 'mark' them as apoptotic.

Lysis of erythrocytes using hypotonic solutions is one approach to remove red blood cells (RBCs) from umbilical cord blood (UCB), bone marrow (BM), and peripheral blood (PB) before flow cytometric analysis or sorting of nucleated cells (NCs). Our team employed this separation step to prepare UCB-, BM-, or PB-derived cells to sort very small embryonic-like stem cells (VSELs). We noticed that depletion of RBCs from UCB by hypotonic lysis resulted in a significant increase in the number of NCs including VSELs that bind Annexin-V (Ann-V). Surprisingly, these cells were not apoptotic and displayed normal proliferative potential. To explain this discrepancy, we show that RBC-derived microvesicles (RMV) released during erythrocyte lysis may transfer phosphatidylserine (PS) to the surface of NCs and 'mark' them falsely positive as apoptotic cells. This observation should be considered whenever Ann-V binding viability assays are employed to evaluate the quality of NCs depleted from erythrocytes via hypotonic lysis.

Very small embryonic-like stem cells are present in adult murine organs: ImageStream-based morphological analysis and distribution studies.

Recently, we purified a population of CXCR4+/Oct-4+/SSEA-1+/Sca-1+/Lin(-)/CD45(-) very small embryonic-like stem cells (VSELs) from adult murine bone marrow (BM). After using flow cytometry, ImageStream analysis, confocal microscopy, and real time RT-PCR, we report that similar cells could be also identified and isolated from several organs in adult mice. The highest total numbers of Oct-4+ VSELs were found in the brain, kidneys, muscles, pancreas, and BM. These observations support our hypothesis that a population of very primitive cells expressing germ line/epiblast markers (Oct-4, SSEA-1) is deposited early during embryogenesis in various organs and survives into adulthood. Further studies are needed to determine whether these cells, after being isolated from various adult human organs similarly to their murine BM-derived counterparts, are endowed with pluripotent stem cell properties.

TGF-beta1 gene polymorphisms and primary vesicoureteral reflux in childhood.

The aim of this study was to assess the association between the transforming growth factor-beta1 (TGF-beta1) gene polymorphisms rs1800469 (commonly known as T-509C) and rs1982073 (commonly known as Leu (10)-->Pro) and primary vesicoureteral reflux (VUR) and renal scarring. Using a case-control approach, we examined 121 children with primary VUR and 169 controls. Genotyping of the TGF-beta1 gene polymorphisms was performed by restriction fragment length polymorphism (RFLP) analysis. The (99m)Tc-DMSA- or (99m)Tc-unitiol-single photon emission computed tomography method was used to evaluate renal scars in 84 of 121 VUR children. Statistical analysis revealed differences in rs1800469 genotype frequencies between VUR patients and controls (p = 0.0021). Our data demonstrate that individuals homozygous for the TT genotype are at risk of primary VUR [odds ratio (95% confidence interval) = 2.7 (1.46-5.08)]. Distribution of the rs1982073 polymorphism was similar in VUR children and controls. In terms of renal scarring, patients were stratified into non-scar and scar subgroups, and no differences in the genotype frequencies of either polymorphism was found. Previous reports have shown that the TT genotype of the rs1800469 polymorphism is a risk factor for renal scarring in primary VUR, and the results of our study suggest that this same polymorphism is associated with susceptibility to this congenital uropathy.

Circadian blood pressure variation in normotensive type 2 diabetes patients and angiotensin converting enzyme polymorphism.

BACKGROUND/AIMS: Loss of circadian blood pressure (BP) variation (i.e., lack of nocturnal BP dip by at least 10mmHg, 'non-dipping') is associated with increased mortality rate in subjects with diabetes. We studied whether angiotensin converting enzyme (ACE) insertion/deletion (I/D) gene polymorphism may play a role in 24-h BP rhythm control. METHODS: The study group was 38 normotensive normoalbuminuric type 2 diabetes patients with impaired BP variation, the controls were 51 well-matched type 2 diabetes subjects with normal 24-h BP rhythm. ACE I/D polymorphism, endothelial function and subclinical inflammation parameters (serum endothelin-1, sE-selectin, intercellular and vascular cell adhesion molecules, tumor necrosis factor-alpha) were assessed. RESULTS: ACE DD genotype was found in 20 (53%), ID genotype in 16 (42%), and II genotype in 2 (5%) study group subjects, while 5 (10%) control subjects had DD genotype, 30 (59%) - ID genotype, and 16 (31%) - II genotype (p<0.0001). Study group subjects presented with marked endothelial dysfunction. CONCLUSION: Impaired circadian blood pressure variation in normotensive normoalbuminuric type 2 diabetes patients is associated with ACE DD genotype and marked endothelial dysfunction when compared to diabetic subjects with normal blood pressure rhythm.

[Polymorphism I/D of the angiotensin-converting enzyme gene and disturbance of blood pressure in type 1 diabetic children and adolescents]. / Zaburzenia cisnienia tetniczego u dzieci i mlodziezy chorej na cukrzyce typu 1 a polimorfizm I/D genu enzymu konwertujacego angiotensyne.

AIM: The aim of the study was to evaluate an association between ACE genotypes and blood pressure (BP) disturbances in children and adolescents withType 1 diabetes mellitus. MATERIALS AND METHODS: 126 normo-albuminuric, type 1 diabetic children and adolescents at the age 10.5-19.7 years, and duration of diabetes 2.0-17.8 years, were included in the study. All patients were clinically normotensive and normoalbuminuric. Twenty-four-hour ambulatory BP monitoring was undertaken in all patients. The values of systolic BP, diastolic BP, mean arterial BP blood pressure and diurnal variation in BP were estimated. Prehypertension was diagnosed as BP values between 90pc or 120/80 mmHg and 95pc. ACE genotypes were assessed using polymerase chain reaction. RESULTS: In 48 individuals (38.1%) prehypertension was diagnosed. ACE genotypes distributed in patients were as follow: 31 (24.6%) genotype II, 45 (35.7%)--ID, and 50 (39.7%)--DD. Patients with DD genotype had higher nocturnal systolic BP (104 vs. 101 mmHg; p=0.029), diastolic BP (54 vs. 52 mmHg, p=0.003) and mean arterial BP (71 vs. 68 mmHg, p=0.003) as compared to carriers of the I allele (group ID + II). In the group DD in compare to the group ID + II lower nocturnal depletion in diastolic BP (18.0 vs. 20.5 mmHg, p=0.024) and mean BP (14.8 vs 16.6 mmHg, p=0.049) was observed. 14% patients of the DD group were non-dipper and 1.32% in the group ID + II (p=0.01). There was no differences in the prevalence of prehypertension between genotype groups (II+ID vs. DD: 38.2 vs. 38.0%, ns). CONCLUSIONS: Genotype DD is associated with nocturnal BP abnormalities in normotensive and normo-albuminuric children and adolescents with type 1 diabetes. There is no relationship between frequency of prehypertension and ACE gene polymorphism.