Serum metabolomics identified specific lipid compounds which may serve as markers of disease progression in patients with Alström and Bardet-Biedl syndromes.

Objectives: Alström syndrome (ALMS) and Bardet-Biedl syndrome (BBS) are among the so-called ciliopathies and are associated with the development of multiple systemic abnormalities, including early childhood obesity and progressive neurodegeneration. Given the progressive deterioration of patients' quality of life, in the absence of defined causal treatment, it seems reasonable to identify the metabolic background of these diseases and search for their progression markers. The aim of this study was to find metabolites characteristic to ALMS and BBS, correlating with clinical course parameters, and related to the diseases progression. Methods: Untargeted metabolomics of serum samples obtained from ALMS and BBS patients (study group; n = 21) and obese/healthy participants (control group; each of 35 participants; n = 70) was performed using LC-QTOF-MS method at the study onset and after 4 years of follow-up. Results: Significant differences in such metabolites as valine, acylcarnitines, sphingomyelins, phosphatidylethanolamines, phosphatidylcholines, as well as lysophosphatidylethanolamines and lysophosphatidylcholines were observed when the study group was compared to both control groups. After a follow-up of the study group, mainly changes in the levels of lysophospholipids and phospholipids (including oxidized phospholipids) were noted. In addition, in case of ALMS/BBS patients, correlations were observed between selected phospholipids and glucose metabolism parameters. We also found correlations of several LPEs with patients' age (p < 0.05), but the level of only one of them (hexacosanoic acid) correlated negatively with age in the ALMS/BBS group, but positively in the other groups. Conclusion: Patients with ALMS/BBS have altered lipid metabolism compared to controls or obese subjects. As the disease progresses, they show elevated levels of lipid oxidation products, which may suggest increased oxidative stress. Selected lipid metabolites may be considered as potential markers of progression of ALMS and BBS syndromes.

Identification of bone metabolism disorders in patients with Alström and Bardet-Biedl syndromes based on markers of bone turnover and mandibular atrophy.

Objectives: Causative variants in genes responsible for Alström syndrome (ALMS) and Bardet-Biedl syndrome (BBS) cause damage to primary cilia associated with correct functioning of cell signaling pathways in many tissues. Despite differences in genetic background, both syndromes affect multiple organs and numerous clinical manifestations are common including obesity, retinal degeneration, insulin resistance, type 2 diabetes and many others. The aim of the study was to evaluate bone metabolism abnormalities and their relation to metabolic disorders based on bone turnover markers and presence of mandibular atrophy in patients with ALMS and BBS syndromes. Material and methods: In 18 patients (11 with ALMS and 7 with BBS aged 5-29) and in 42 age-matched (p < 0.05) healthy subjects, the following markers of bone turnover were assessed: serum osteocalcin (OC), osteoprotegerin (OPG), s-RANKL and urinary deoxypyridinoline - DPD. In addition, a severity of alveolar atrophy using dental panoramic radiograms was evaluated. Results: Lower serum OC (p = 0.0004) and urinary DPD levels (p = 0.0056) were observed in the study group compared to controls. In ALMS and BBS patients, serum OC and urinary DPD values negatively correlated with the HOMA-IR index, while a positive correlation between the OC and 25-OHD levels and a negative correlation between s-RANKL and fasting glucose concentrations were found. A significant difference in the incidence of low-grade mandibular atrophy between patients with ALMS and BBS and controls (p < 0.0001) was observed. Conclusions: The identification of bone metabolism disorders in patients with ALMS and BBS syndromes indicates the necessity to provide them with appropriate diagnosis and treatment of these abnormalities.

The Role of Noise in Specific Detectivity of InAs/GaSb Superlattice MWIR Bariodes.

In this paper, the results of the electrical, noise, and optical characterization of p-i-n and p-B-i-n diodes with AlSb and 4 ML AlSb/8 ML GaSb superlattice barriers in High-Operating Temperature conditions, are presented. Experimental and theoretical noise parameters were compared. Both dark current and noise analysis showed that the p-Bp_bulk-i-n bariode had the best performance. P-i-n photodiodes had the highest experimental value of specific detectivity (D*) of 6.16 × 109 Jones at 210 K and zero bias. At about -1 V reverse bias, the bariode with AlSb/GaSb electron barrier caught up to it and both devices achieved D* = (1-1.1) × 108 Jones. Further optimization of the superlattice-based electron barrier should result in the improvement of bariode performance at a smaller bias, at which better noise performance is more pronounced. It was shown that neglecting the low-frequency noise component can lead to a significant overestimation of detectivity. The simple method of incorporation of low-frequency noise contribution in the detectivity calculation, without time-consuming measurements, has been proposed.

Conjunctival Ultraviolet Autofluorescence as a Measure of Riboflavin and Ultraviolet and Accelerated Cross-Linking Exposure in Keratoconic Patients.

PURPOSE: The study was performed to analyze the prevalence of the conjunctival ultraviolet autofluorescence (CUVAF) area in keratoconic eyes and changes caused by UVA-irradiation as a component of accelerated corneal cross-linking (aCXL). METHODS: The study group involved 20 keratoconic patients subjected to aCXL surgery in one eye. The comparative group consisted of 111 age- and sex-matched patients with healthy corneas. The images of the anterior segment in both patient groups were taken using a Coroneo camera. In the study group the photos were taken before and immediately after the surgery, and 7 and 30 days following the procedure. RESULTS: Nasal and temporal autofluorescence area (AN+T) were significantly smaller in a keratoconic patients group compared to control group (p = 0.0001). Patients with the third stage of keratoconus had significantly higher AN+T (p = 0.0277) compared with individuals with lower stage keratoconus. No statistically significant CUVAF changes were observed after the aCXL procedure. In keratoconic patients with primary CUVAF undergoing aCXL, a temporary fast enlargement of the autofluorescence area was observed. CONCLUSIONS: The eyes undergoing the aCXL procedure showed no difference in the size of the CUVAF area but such patients should be in strict follow-up in order to reveal UV-related ocular surface diseases.

Central Corneal Thickness can be Related to Diabetic Peripheral Neuropathy in Children with Type 1 Diabetes.

AIMS: Diabetic eye disease with its various manifestations as well as diabetic neuropathy may occur in patients with type 1 diabetes (T1D) after several years of diabetes duration. Pachymetry is a promising method evaluating central corneal thickness (CCT) in diabetic patients. The aim of the study was to evaluate the CCT values in children with T1D and its relationship to neurophysiological markers of diabetic neuropathy. METHODS: The study groups included 119 T1D children with average 5.3 years of diabetes duration and 38 age-matched controls. CCT index was measured with pachymeter in all subjects and in 19/119 of T1D patients the CCT values were referred to the ENG-EMG-SSR study results. RESULTS: In T1D patients the higher CCT values were observed as compared to healthy controls (p=0.037). Correlations between CCT values and both distal latency of the motor fibers of the median nerve (R=0.51; p=0.044) and conduction velocity of this nerve (R=-0.55; p=0.027) were noted. A conduction velocity of the sensory fibers of sural nerve correlated negatively with CCT index (R=-0.50; p=0.045) in the T1D patients. CONCLUSIONS: CCT measurement may be helpful in the referral of the asymptomatic pediatric T1D patients to assess an early stage of diabetic neuropathy.

Evaluation of Size and Location of a Mental Foramen in the Polish Population Using Cone-Beam Computed Tomography.

INTRODUCTION: The mental foramen (MF) is a bilateral opening localized on an anterior surface of the mandible. A precise location as well as well-defined shape, size, and number of the MF is crucial for different clinical dental procedures. The aim of this study was to determine a size and location of the MF in relation to the lower teeth using the cone-beam computed tomography (CBCT) study. MATERIAL AND METHODS: In a group of 201 patients (106 males and 95 females) the CBCT images were performed using the GX CB-500 device (Gendex, USA). RESULTS: No significant differences in values of the horizontal (H) and vertical (V) diameters as well as the H:V ratio on both sides in relation to the age of participants were found. In males both average values of a horizontal diameter (p=0.031) and vertical diameter (p=0.001) were significantly higher on the right side than in the female subgroup, whereas on the left side only an average value of a vertical diameter was significantly higher in men (p=0.006) in comparison to women. Moreover, the H:V ratio was significantly lower in males on the left side (p=0.032). There were no significant relationships between age and gender of the patients (p>0.05) and the type of mental foramen on the right and left sides. CONCLUSIONS: The application of the CBCT study enabled a precise determination of the shape, size, and position of the mental foramen in relation to the neighboring anatomical structures on a representative group of the Polish patients. The results obtained may contribute to guidelines for dental procedures including anesthesia of the mental nerve and endodontic, implantology, and dental surgery with regard to the location of mental foramen depending on the sex and age of patients.

The assessment of accessory mental foramen in a selected polish population: a CBCT study.

BACKGROUND: Accessory mental foramen (AMF) is a rare anatomical variation. When accessory mental foramen is present, the nerves and vessels that go through the mental foramen (MF) must follow alternative courses and special care must be taken during dental treatment planning. The purpose of this study was to evaluate the occurrence and the location of AMF in a selected Polish population using cone-beam computed tomography (CBCT). METHODS: Two hundred CBCT (105 males and 95 females) examinations were evaluated for the presence of AMFs. The location and side of AMFs were reported. The mean distance between MF and AMF was also calculated. The vertical size of MF on the side with and without AMF was measured. The obtained variables were statistically analyzed. RESULTS: AMFs were observed in 7% of the patients. There was no statistically significant difference between the appearance of AMF and sex (p > 0.05). We found no significant difference in the vertical size of MF between individuals with and without AMFs (p < 0.05). CONCLUSION: Twenty-eight AMFs (7%) were observed from 400 sides of 200 patients. AMFs occurred more often in males (18 AMFs) than in females (10 AMFs). Twenty AMFs (71.4%) were located anteriorly, and eight (28.6%) - posteriorly. Fifteen AMFs (53.6%) were on the right side and thirteen (46.4%) - on the left.

Decreased FOXP3 mRNA expression in children with atopic asthma and IgE-mediated food allergy.

BACKGROUND: The role of T regulatory lymphocytes has been investigated in various allergic diseases. However, the precise relation between the phenotype and severity of allergic diseases and the changes in FOXP3 mRNA expression are not fully understood. OBJECTIVE: To compare the expression of FOXP3 mRNA in children with asthma with and without concomitant food allergy (FA) with healthy children and children with only FA. METHODS: The study included 82 children: 15 with atopic asthma and IgE-dependent FA, 27 with atopic asthma without FA, 20 with IgE-dependent FA without asthma, and 20 healthy children without atopy. Reverse transcription was performed using a commercially available High Capacity cDNA Archive Kit (Applied Biosystems, Carlsbad, California). Analysis was carried out with a 7900HT real-time polymerase chain reaction system (Applied Biosystems). RESULTS: The average level of the FOXP3 gene expression in children with allergy was significantly lower compared with healthy children (2.2 ± 1.3 vs 4.2 ± 4.2; P = .014). The lowest mean level of FOXP3 mRNA expression (1.9 ± 1.6) was recorded in children with asthma and FA, and the highest level (4.2 ± 4.2) was recorded in healthy children without atopy (P = .036). A milder course of asthma or the degree of allergic reaction after a food challenge was associated with higher FOXP3 mRNA expression. CONCLUSION: Significantly lower levels of FOXP3 gene expression, observed more commonly in children with asthma and IgE-dependent FA than in healthy controls, were associated with a more severe clinical course. Therefore, FOXP3 expression could serve as an indicator of severe asthma with concomitant atopic conditions such as IgE-dependent FA.

The effect of IL-2, IL-12 and IL15 on the function of natural killer cells in children suffering from type 1 diabetes mellitus.

INTRODUCTION: The immunologic reaction of pancreatic islets destruction leads to the occurrence of type 1 diabetes mellitus (T1D). The autoreactive lymphocytes play the pivotal role in this process although mechanisms regulating the lymphocyte migration and infiltration of Langerhans islets have not been fully understood yet. The in vitro studies showed natural killer (NK) cells potency to initiate pancreatic islets cell lyses. Many authors postulate that NK cells may be involved in this reaction. AIM OF THE STUDY: The aim of the study was to evaluate the effect of IL-2, IL-12 and IL-15 stimulation on peripheral blood NK cells in children suffering from type 1 diabetes mellitus in comparison to healthy controls. MATERIAL AND METHODS: Fifteen children with type 1 diabetes and 10 healthy adults were examined. NK cells were isolated by the magnetic cell separation system (MACS). For activation, NK cells were cultured with IL-2, IL-12 and IL-15 for 24 hours. The production of IFN-γ and IL-10 by NK cells was measured using commercial ELISA kits. FACS analysis of cell surface antigens--CD16, CD56, NKG2D and CD137 was performed using LSR II flow cytometer. RESULTS: In children with T1D the IFN-γ median concentration in supernatant obtained from NK cells culture was 16.831 ng/ml (inter quartile range 5.566-25.509) and did not statistically differ from median IFN-γ concentration in the control group--14.810 ng/ml (7.022-18.785), p = 0.76. In contrast, the IL-10 median concentration was statistically higher in T1D patients 7.87 pg/ml (1.32-11.37) than in healthy participants--1.41 pg/ml (1.05-4.81), p = 0.01. The median (inter-quartile range) percentage of NK NKG2D(+) was found in 0.42% (0.28-0.76) cells of TID patients versus 0.72% (0.53-1.08) in the controls (p = 0.05). There was no difference between -T1D group and the control group in regard to NK cells expressing CD137 - 6.58% (3.38-12.4) versus 6.85% (2.94-10.8); p = 0.8. CONCLUSIONS: The observed activity of NK cells after in vitro stimulation by IL2, IL-12 and IL15 in children suffering from type 1 diabetes mellitus indicates the tendency for supporting the inhibition of autoimmunological reaction by increased IL10 synthesis and increased number of NK cells with surface NKG2D receptors.

Age-dependent changes of serum soluble CD30 concentration in children.

CD30 was originally described as a marker on Reed-Sternberg cells in Hodgkin lymphoma. The extracellular portion of CD30 is proteolytically cleaved from CD30+ cells, to produce a soluble form of the molecule (sCD30) detectable in serum. Measurement of sCD30 concentration in serum has been suggested to be a potential tool in monitoring of inflammatory status in variety of diseases. Several investigators reported the relevance for sCD30 as a predictive marker for allograft rejection following organ transplantation. The aim of the study was to verify whether sCD30 serum concentrations may be affected by an age in healthy children. Heparinized venous blood was taken from 78 healthy children. For the analysis of sCD30 levels, the commercially available sCD30 ELISA was used. The sCD30 was detected in all serum samples and concentrations ranged from 6.75 to 68.07ng/mL. The statistical analysis of all individuals showed that sCD30 concentration was significantly age depended (r=-0.618, p<0.0001). When sCD30 concentrations were analyzed in regard to gender, no significant differences were identified in age subgroups.

The 5'VNTR proinsulin gene (INS) polymorphism and the functional reserve of ß cells in the intravenous glucose tolerance test.

INTRODUCTION: Proinsulin 5'VNTR polymorphism determines susceptibility to type 1 diabetes (T1DM). The authors tested whether it affects intravenous glucose tolerance test (IVGTT) results. AIM OF THE STUDY: To evaluate a possible relationship between 5'VNTR proinsulin gene (INS) polymorphism and glucose, insulin and C-peptide levels during IVGTT among siblings of children suffering from T1DM. MATERIAL AND METHODS: Fourteen patients - siblings of children with type 1 diabetes, positive for at least one autoantibody, underwent IVGTT with glucose, insulin and C-peptide concentrations measurement. RESULTS: Mean age of patients equaled 10.71 ± 4.15 years. Eight individuals were homozygous for class I/I and six were class III/I heterozygotes. No significant differences in blood glucose levels during the IVGTT were observed (p=0.67). However, lower insulin (p=0.03) and C-peptide (p=0.01) levels were observed in I/I homozygotes in post-challenge timepoints. No significant differences were observed in baseline fasting insulin, glucose and C-peptide levels. CONCLUSIONS: The class III allele in the 5'VNTR promoter region of INS is associated with a greater functional reserve of ß cells in response to a direct hyperglycemic stimulus in individuals with a familial background of T1DM.

Effects of 1-methylnicotinamide and its metabolite N-methyl-2-pyridone-5-carboxamide on streptozotocin-induced toxicity in murine insulinoma MIN6 cell line.

1-methylnicotinamide (MNA) is a primary metabolite of nicotinamide. In recent years several activities of MNA have been described, such as anti-inflammatory activity in skin diseases, induction of prostacyclin synthesis via COX-2, aortal endothelium protection in diabetes and hypertriglyceridaemia and increased survival rate of diabetic rats. 1-methylnicotinamide was also suggested to protect pancreatic cells from streptozotocin in vivo. Streptozotocin toxicity is known to be mediated by poly-ADP-ribose polymerase. Nicotinamide and its derivatives have been shown to ameliorate poly-ADP-ribose polymerase-dependent nucleotide pool reduction. We aimed to verify if 1-methylnicotinamide and its metabolite, N-methyl-2-pyridone-5-carboxamide, can protect insulinoma cells from streptozotocin-induced toxicity. We found that N-methyl-2-pyridone-5-carboxamide, but not 1-methylnicotinamide, restores the pool of ATP and NAD+ in streptozotocin-treated cells, but neither compound improved the cell viability. We conclude that inhibition of poly-ADP-ribose polymerase-dependent nucleotide pool reduction may not be sufficient to protect cells from streptozotocin toxicity.

FOXP3, IL-10, and TGF-ß genes expression in children with IgE-dependent food allergy.

BACKGROUND: Regulatory T cells (Tregs) have an essential role in tolerance and immune regulation. However, few and controversial data have been published to date on the role and number of these cells in food allergic children. The forkhead/winged-helix transcription factor box protein 3 (FOXP3) is considered the most reliable marker for Tregs. OBJECTIVE: This study aims to investigate the FOXP3, interleukin (IL)-10, and transforming growth factor (TGF-ß) genes expression in children with IgE-dependent food allergy. MATERIAL AND METHODS: The study group consisted of 54 children with IgE-dependent food allergy (FA) and a control group of 26 non-atopic healthy children. The diagnosis of FA was established using questionnaires, clinical criteria, skin prick tests, serum sIgE antibodies (UniCAP 100 Pharmacia Upjohn), and a double-blind placebo control food challenge. In order to assess gene expression, the isolation of nucleated cells was performed using Histopaque-1077 (Sigma-Aldrich, Germany). The concentration of RNA obtained was measured using a super-sensitive NanoDrop ND1000 spectrophotometer (Thermo Scientific, USA). A reverse transcription reaction was performed using a commercially available set of High Capacity cDNA Archive Kit (Applied Biosystems, USA). Analysis have been carried out in the genetic analyzer 7900HT Real-Time PCR (Applied Biosystems, USA). RESULTS: The average level of the FOXP3 gene expression in the studied group was 2.19 ± 1.16 and in the control group 2.88 ± 1.66 (p = 0.03). The average level of IL10 mRNA expression in the study group was 13.6 ± 1.07 and was significantly lower than corresponding values in the control group 14.3 ± 1.1 (p = 0.01). There were no significant differences in the average level of the TGF-ß mRNA expression in the study group (3.4 ± 0.4) and controls (3.5 ± 0.3; p > 0.05). The FOXP3 gene expression was the highest in children who acquired tolerance to food (3.54 ± 0.75), lower in heated allergen-tolerant children (2.43 ± 0.81), and the lowest in heated allergen-reactive children (1.18 ± 0.5; p = 0.001 control vs heated allergen reactive; p = 0.005 heated allergen tolerant vs heated allergen reactive; p = 0.001 outgrown vs heated allergen reactive). The significant tendency toward lower total IgE levels with a higher FOXP3 mRNA expression was detected (n = 54; Pearson r = -0.4393; p = 0.001). CONCLUSIONS: Children with FA showed statistically significant lower level of the FOXP3 and IL10 gene expression than healthy children. Children acquiring tolerance to the food show significantly higher levels of the FOXP3 gene expression than children with active FA. The correlation between the level of FOXP3 and total IgE was detected.

Evaluation of receptor expression on immune system cells in the peripheral blood of asthmatic children undergoing food challenges.

BACKGROUND: The role of food allergens in the induction of allergic reactions in the airways is not completely understood. The aim of the present study was to evaluate fluorocytometric assays of the peripheral blood during food challenge tests in children with asthma and food allergy. PATIENTS AND METHODS: 22 children with asthma and concomitant food allergy and 18 children with asthma without food allergy participated in the study. Oral challenge tests were performed using double-blind, placebo-controlled food challenge. Blood samples were collected before and 4 and 24 h after the challenge. CD25 and CD23 antigen expression was determined with monoclonal antibodies using a FACSCalibur flow. RESULTS: The evaluation of the CD25+ T subpopulation and CD19+CD23+ B lymphocytes revealed statistically significant differences between the study group and the control group. In children with asthma and food allergy, the cell pool consisted (on average) of 9 +/- 2.8% of CD3+CD25+ cells before the challenge and of 10.3 +/- 3.8% (mean delta: 1.623; p = 0.01) after the provocation. However, placebo challenge did not significantly change the number of this T-lymphocyte subpopulation (mean delta: -0.121; p > 0.05). The highest increase in the CD25+ T-subpopulation expression was found in patients with respiratory reactions during the positive food challenge (mean delta: 4.065; p < 0.004). CONCLUSIONS: An increase in CD25+ T-lymphocyte and CD23 B-lymphocyte populations after food allergen challenge may indicate their significant role in the pathogenesis of the active phase of the immunoinflammatory process in children with asthma and concomitant food allergy.

Comparative studies of lymph node cell subpopulations and cytokine expression in murine model for testing the potentials of chemicals to induce respiratory sensitization.

OBJECTIVES: To investigate immunological changes in lymph nodes based on expression of cell-specific receptors and cytokine expression profile and accompanying inflammatory reactions in lungs of mice treated with chemicals of known potentials to induce respiratory sensitization and those in which activity in this regard is unclear. MATERIALS AND METHODS: On day 1 and 7, Balb/c mice received toluene-2,4-diisocyanate (TDI), trimellitic anhydride (TMA), 1-chloro-2,4-dinitrobenzene (DNCB), glutaraldehyde (GA), formaldehyde (FA), benzalkonium chloride (ChB) or vehicle. On day 14, they received a single intranasal instillation with the same chemical or vehicle. On day 15, auricular lymph nodes (LN) were excised and used for analyzes of T-, B-cells, expression of CD44 and for the estimation of IL-4 and IFN-gamma production after in vitro stimulation with concanavalin A (ConA) and also for IL-4 and IFN-gamma mRNA expression analyses using Real-Time PCR. Inflammatory changes in lungs were observed by estimation of TNF-alpha and MIP-2 concentrations and cell numbers and their type in BAL. RESULTS: There were no significant changes in cell subpopulations of T helper cells in LN. The percent of B cells was significantly increased after treatment with DNCB, TDI, and GA. Increased expression of CD44 on T cells was also observed. Both IL-4 and IFN-gamma were found increased in TDI- and FA-treated mice, while only IL-4 was increased in TMA-treated mice. Real-Time PCR analyses, however, showed increased IL-4 mRNA expression for TDI- and TMA-, and IFN-gamma mRNA expression for DNCB-treated mice. We haven't observed significant changes in inflammatory reactions in the lungs of exposed animals. CONCLUSIONS: Studying immunological changes with first determining the activation status of T cells followed by analyzes of expression of mRNA for Th1 and Th2 cytokines in murine model could be a useful method for assessment of the potentials of chemicals to induce respiratory sensitization but is not sufficient. Addition of ventilatory measurements, but not necessarily inflammatory reactions, could complete the model.

Influence of interleukin IL-2 and IL-12 + IL-18 on surface expression of immunoglobulin-like receptors KIR2DL1, KIR2DL2, and KIR3DL2 in natural killer cells.

Natural killer (NK) cells express killer cell inhibitory receptors (KIRs) that recognize polymorphic class I MHC molecules. In the present study, we analyze the modulatory effect of IL-2 alone or a combination of IL-12 with IL-18 on surface expression of killer cell immunoglobulin-like receptors KIR2DL1, KIR2DL2, and KIR3DL2 in NK cells. Thus, it was found that IL-2 causes a significant increase in the proportion of cells with given studied receptors. Stimulation by a mixture of IL-12 and IL-18 caused significant increase in the fraction of cells with the KIR2DL1 and KIR2DL2, however no significant change in the percentage of cells with KIR3DL2 receptor on their surface was observed. The results of the study show the presence of KIRs on both resting and activated NK cells, this may suggest that KIRs have also an important role in the regulatory processes after activation of this subpopulation of cells.