Krystyna Wyka

The cross-reactivity of anti-asparaginase antibodies against different L-asparaginase preparations.

Repeated administration of l-asparaginase leads to the development of specific antibodies and hypersensitivity reactions. The aim of the study was to evaluate a possible cross-reaction of anti-asparaginase antibodies, developed against the native E. colil-asparaginase (Asparaginase Medac), with other preparations of the enzyme. Sixteen patients with acute lymphoblastic leukemia, in whom in the reinduction phase of treatment hypersensitivity against l-asparaginase was observed and/or the presence of anti-asparaginase antibodies was established were recruited for the present study. Ten out of 16 tested sera showed cross-immunoreactivity to PEG-asparaginase, while no reactivity to l-asparaginase derived from Erwinia chrysantemi was observed. Since cross-reacting antibodies were also found in sera of patients with no overt allergic reaction, l-asparaginase may undergo silent inactivation during the reinduction phase of therapy. This finding is of clinical importance with regard to appropriate dosage and necessitates careful enzyme activity monitoring in all patients undergoing repeated treatment with various l-asparaginase preparations.

SERUM LEVELS OF VASCULAR ENDOTHELIAL GROWTH FACTOR AND BASIC FIBROBLASTIC GROWTH FACTOR IN CHILDREN WITH HEMANGIOMAS AND VASCULAR MALFORMATIONS--PRELIMINARY REPORT

Abstract:  Impaired balance between proangiogenic and antiangiogenic factors has been implicated in the development of hemangiomas. Elevated vascular endothelial growth factor serum levels and basic fibroblastic growth factor urine levels in patients with proliferating hemangiomas were reported. However, whether these growth factors can be used for the differential diagnosis of vascular anomalies or assessment of the clinical course of hemangiomas has yet to be determined. We report here our preliminary results of serum vascular endothelial growth factor and basic fibroblastic growth factor levels as an aid in the diagnosis of hemangiomas and in the follow up of patients with this lesion. Twenty two children with infantile hemangioma (13 with proliferating hemangiomas, nine with involuting lesions), five children with vascular malformations, and 25 healthy children study group. Vascular endothelial growth factor and basic fibroblastic growth factor serum levels were analyzed by an ELISA assay. The serum vascular endothelial growth factor concentrations in children with proliferating hemangiomas were significantly higher than in patients with involuting hemangiomas, vascular malformations and healthy patients. The serum basic fibroblastic growth factor concentrations were low and similar in all patients with no statistical correlation between study groups. We concluded that (i) ELISA can easily determine vascular endothelial growth factor concentrations in different phases of hemangioma growth and help distinguishing them from vascular malformations. (ii) A potential role for vascular endothelial growth factor in the pathophysiology of hemangiomas is probable.

Mutations in the ABCC8 (SUR1 subunit of the KATP channel) gene are associated with a variable clinical phenotype

Objective  Mutations in the ABCC8 gene encoding the SUR1 subunits of the β‐cell K‐ATP channel cause neonatal diabetes (ND) mellitus. We aimed to determine the contribution of ABCC8 gene to ND in Poland, to describe the clinical phenotype associated with its mutations and to examine potential modifying factors.

HDL cholesterol as a diagnostic tool for clinical differentiation of GCK‐MODY from HNF1A‐MODY and type 1 diabetes in children and young adults

Introduction  Confirmation of monogenic diabetes caused by glucokinase mutations (GCK‐MODY) allows pharmacogenetic intervention in the form of insulin discontinuation. This is especially important among paediatric and young adult populations where GCK‐MODY is most prevalent.

Interleukin 18 as a Marker of Chronic Nephropathy in Children after Anticancer Treatment

Novel markers of nephrotoxicity, including kidney injury molecule 1 (KIM-1), interleukin 18 (IL-18), and beta-2 microglobulin, were used in the detection of acute renal injury. The aim of the study was to establish the frequency of postchemotherapy chronic kidney dysfunction in children and to assess the efficacy of IL-18, KIM-1, and beta-2 microglobulin in the detection of chronic nephropathy. We examined eighty-five patients after chemotherapy (median age of twelve years). The median age at the point of diagnosis was 4.2 years, and the median follow-up time was 4.6 years. We performed classic laboratory tests assessing kidney function and compared the results with novel markers (KIM-1, beta-2 microglobulin, and IL-18). Features of subclinical renal injury were identified in forty-eight children (56.3% of the examined group). Nephropathy, especially tubulopathy, appeared more frequently in patients treated with ifosfamide, cisplatin, and/or carboplatin, following nephrectomy or abdominal radiotherapy (P = 0.14, P = 0.11, and P = 0.08, resp.). Concentrations of IL-18 and beta-2 microglobulin were comparable with classic signs of tubulopathy (P = 0.0001 and P = 0.05). Concentrations of IL-18 were also significantly higher in children treated with highly nephrotoxic drugs (P = 0.0004) following nephrectomy (P = 0.0007) and abdominal radiotherapy (P = 0.01). Concentrations of beta-2 microglobulin were higher after highly toxic chemotherapy (P = 0.004) and after radiotherapy (P = 0.02). ROC curves created utilizing IL-18 data allowed us to distinguish between children with nephropathy (value 28.8 pg/mL) and tubulopathy (37.1 pg/mL). Beta-2 microglobulin and IL-18 seem to be promising markers of chronic renal injury in children after chemotherapy.

Assessment of interleukin-6, interleukin-8 and interleukin-18 count in the serum of IUGR newborns

Abstract Aim: Aim of this study was to assess concentrations of interleukin-6 (IL-6), interleukin-8 (IL-8) and interleukin-18 (IL-18) in the serum of newborns with diagnosed intrauterine growth restriction (IUGR) in comparison to concentrations in serum of newborns with weight appropriate for gestational age (AGA). Materials: Research was conducted at the Lodz Medical University Clinic of Neonatology during 2010–2011. Surveyed group consisted of 50 hypotrophic full-term infants of single pregnancies (average weight: 2329 ± 287 g); control group, enclosing 50 infants AGA (average weight: 3544 ± 2161 g). Both groups received average Apgar score of 9 points. Concentrations of analysed cytokines were marked between 4–6 hours after birth. The enzyme-linked immunosorbent assay (ELISA) test was used to determine interleukins concentrations. Study was prospective. Statistics on the data were conducted with the Kolmogorov–Smirnov test. Significance level: p < 0.05. Results: Concentrations of IL-6 and IL-18 were elevated in the IUGR group in a statistically significant manner in comparison to the control group. Conclusions: An elevated level of IL-6 and IL-18 in the IUGR group, comparing to control group, signifies the existence of inflammation in the process of developing IUGR, therefore, screening tests estimating levels of interleukins as IL-6 and IL-18 might be clinically useful in predicting the occurrence of IUGR and help preventing it.

Population-based estimates for double diabetes amongst people with glucokinase monogenic diabetes, GCK-MODY

Monogenic diabetes has become an increasingly active area of research in recent years, allowing for the establishment of several large registries and epidemiologic estimates [1–3]. In the paediatric population, mutation of the glucokinase gene (GCK) is most frequently responsible for the development of monogenic diabetes, typically characterized by low HbA1c levels and low risk of complications [2]. However, we have observed patients with poor metabolic control in the Polish GCK maturity-onset diabetes of the young (MODY) cohort, and hypothesized that such individuals may have a second cause of diabetes. To calculate the expected number of patients with different overlapping types of diabetes, we used data from available epidemiologic reports [1–3] and the Central Statistical Office of Poland (www.stat.gov.pl/bdlen/ app/strona.html?p_name-indeks). To estimate the frequency of this ‘double diabetes’, we assumed the frequency of monogenic diabetes to equal 108 per million individuals [1], out of whom approximately one third were expected to have GCK-MODY. The overall prevalence of diabetes in Poland was estimated using the International Diabetes Federation (IDF) Diabetes Atlas [4]. To cover the whole population from 0 to 79 years, we added prevalence data on paediatric patients described in earlier epidemiologic reports [2,5] to the IDF estimates (which covered ages 20–79 years). The distribution of major types of diabetes was assumed to be 90% Type 2 and 10% Type 1 diabetes [6]. As there was no evidence of monogenic diabetes being protective against either Type 1 or Type 2 diabetes, we multiplied the probability of having these types by the number of individuals with monogenic diabetes. Patient recruitment and data collection in the group of individuals with GCK-MODY was described in earlier reports [7–9] and in the Supporting Information (Appendix S1). The study was approved by the Bioethics Committee of the Medical University of Lodz. All study participants gave their informed consent. In Poland, 1.54 million (4.0%) individuals have diabetes [4]. Based on earlier estimates, the total number of patients with monogenic diabetes would equal 4162 (95% CI 4038–4290). The number of patients with Type 2 diabetes overlapping on monogenic diabetes in Poland should thus reach 150 (3.6%, 95% CI 3.1–4.3). Similarly, 17 individuals with concomitant Type 1diabetes and monogenic diabetes would be expected from such an estimate [0.4% (95% CI 0.3–0.6)]. To determine how many current patients with such overlapping phenotypes there are, we evaluated individuals in the Polish Nationwide Registry of Monogenic Diabetes with GCKMODY, as its distinctive phenotype marked out individuals with highly unusual phenotype characteristics. Of the 285 eligible patients, eight individuals (2.8%) had HbA1c levels > 59 mmol/mol (7.5%; see also Supporting Information, Fig. S1). Seven of them had shown features of Type 2 diabetes at the time of diagnosis (Table 1), were more likely to be obese [odds ratio (95% CI): 11.78 (1.90–79.27)], more likely to manifest polyuria/polydipsia [odds ratio (95% CI): 5.92 (0.96–6.7)] and required insulin treatment more frequently [odds ratio (95% CI): 20.64 (2.46–173.10)] than the rest of the group with GCK-MODY. BMI status was significantly associated with HbA1c level (Fd.f.=3 = 103.75; P < 0.0001); 31 (10.88%) patients were overweight and 11 (3.95%) were obese. However, obesity alone did not seem sufficient to explain high HbA1c. Out of 11 individuals with BMI > 30 kg/m, only two had HbA1c levels > 59 mmol/mol. This suggested that an additional factor, such as insulin resistance or impaired b-cell function, was present in the seven individuals with Type 2 diabetes-like phenotype and GCK-MODY. The eighth patient suspected for double diabetes showed features of Type 1 diabetes. The probability of monogenic diabetes computed with the MODY calculator [10] for the eight identified patients was well below the threshold recommended for the UK population. This confirmed that, had the patients not been identified as relatives of children previously screened for GCK-MODY, or not developed diabetes in childhood, they would not be considered eligible for genetic testing. Considering the circumstances in which patients with double diabetes were enrolled into the genetic screening programme, it seems well justified that a paediatric-centred genetic screening is a feasible solution that would not exclude them on the basis of metabolic control and traits uncommon to monogenic diabetes. The HbA1c threshold of 59 mmol/mol (7.5%) used in our search for atypical GCK-MODY was selected because of its importance in guidelines for MODY diagnosis and current screening programmes. The observed number of seven patients with Type 2 and GCK-MODY diabetes identified through our search was in line with a recent paper by Steele et al., who showed that six out of 235 individuals with GCK-MODY had high HbA1c levels [66–79 mmol/mol (8.2–9.4%)], accompanied by traits of Type 2 diabetes [11]. Both frequencies are lower than the expected 3.6% based on prevalence of Type 2 diabetes, but the difference may result from more aggressive treatment or relatively young age of the cohort. It is, however, evident that such a therapeutic challenge will arise as more and more patients with various types of monogenic diabetes are identified using novel molecular methods. As therapeutic trials on such patients are unlikely, it seems reasonable to focus on achievement of metabolic goals in patients with double diabetes, even if this mandates treatment intensification not routinely recommended in GCK-MODY. In conclusion, the coexistence of a second type of diabetes with GCK-MODY may hinder recruitment to genetic screening programmes and may necessitate a more aggressive therapeutic approach.

Neonatal diabetes in a child positive for islet cell antibodies at onset and Kir6.2 activating mutation

In contrast to the autoimmune type 1 diabetes, patients with monogenic diabetes due to KCNJ11 mutations do not have pancreatic auto-antibodies at onset. Here we describe a patient diagnosed at the age of 12 weeks that showed ICA at diagnosis, yet has been tested with positive result for KCNJ11 mutation.

Genetic Variability of GCKR Alters Lipid Profiles in Children with Monogenic and Autoimmune Diabetes

PURPOSE Mutations in the glucokinase (GCK) gene are associated with altered blood glucose and lipid concentrations. Our aim was to assess the effects on HbA1c and serum lipid levels of single nucleotide polymorphisms (SNPs) in 2 genes encoding proteins that interact with glucokinase: glucose-6-phospatase catalytic subunit 2 (G6PC2) and glucokinase regulatory protein (GCKR). METHODS The study group included 129 children with GCK-MODY from the Polish Registry of Monogenic Diabetes and 395 with type 1 diabetes (T1DM), in whom we genotyped 2 SNPs in G6PC2 (rs560887) and GCKR (rs1260326). Lipid concentrations were assessed in fasting serum samples. RESULTS Total and HDL cholesterol concentrations were significantly lower in the GCK-MODY group than in patients with T1DM (167.5±32.5 mg/dl vs. 174.4±31.1 mg/dl, p=0.0435 and 48.42±14.3 mg/dl vs. 58.7±12.7 mg/dl, p<0.0001, respectively). No differences in genotype distributions were found except for underrepresentation of GCKR TT homozygotes among GCK-MODY patients (10.9% in GCK-MODY vs. 17.7% in T1DM, p=0.0651). GCKR genotypes showed significant associations with lipid profiles and HbA1c levels, whereas no such associations were noted for G6PC2. After adjustment for confounders, TT homozygotes were shown to have higher total cholesterol and marginally higher LDL cholesterol and triglyceride levels (p=0.0245, p=0.0657 and p=0.0550, respectively). The difference between TT homozygotes and other genotypes was similar in magnitude within the GCK-MODY and T1DM groups. No significant interactions between the type of diabetes and the GCKR or G6PC2 genotype were detected. CONCLUSIONS Individuals who are homozygous TT at rs1260326 of the GCKR gene have higher triglyceride, total and LDL cholesterol levels regardless of the presence of GCK mutations.

Chromosome 18q deletion syndrome with autoimmune diabetes mellitus: putative genomic loci for autoimmunity and immunodeficiency.

A girl with 18q deletion syndrome was diagnosed with autoimmune diabetes mellitus and Hashimotos thyroiditis at the age of 3 yr. In addition, the girl suffered from recurrent infections due to immunoglobulin A and IgG4 deficiency. She was also found to have CD3+CD4+FoxP3+, CD3+CD4+FoxP3+CD25+, and CD3+CD4+CD25+CD127 regulatory T cells deficiency. The exceptional coincidence of the two autoimmune disorders occurring at an early age, and associated with immune deficiency, implies that genes located on deleted 19.4 Mbp region at 18q21.32‐q23 (chr18:58,660,699‐78,012,870) might play a role in the pathogenesis of autoimmunity leading to β cell destruction and diabetes.