Clinical Features and Outcomes of 23 Patients with Wiskott-Aldrich Syndrome: A Single-Center Experience

Objective: Wiskott-Aldrich syndrome (WAS) is an X-linked primary immune deficiency characterized by microthrombocytopenia, eczema, and recurrent infections. We aimed to evaluate the clinical features and outcomes of a WAS cohort. Materials and Methods: We retrospectively evaluated the clinical courses, immunological features, treatments, and outcomes in a total of 23 WAS patients together with data related to 11 transplanted cases among them between 1982 and 2019. Results: Before admission, 11 patients (48%) were misdiagnosed with immune thrombocytopenia. WAS scores were mostly 4 or 5. Eleven patients were transplanted and they had an overall survival rate of 100% during a median follow-up period of 8.5 years (range: 8 months to 20 years). Five patients who were not transplanted died at a median of 7 years (range: 2-26 years). Nontransplanted patients had high morbidity due to organ damage, mostly caused by autoimmunity, bleeding, and infections. Two novel mutations were also defined. Conclusion: All male babies with microthrombocytopenia should be evaluated for WAS. Hematopoietic stem cell transplantation should be performed at the earliest age with the best possible donors.

Evaluation of Low and High Surface Area TiO2 and Al2O3 Metal Oxides-Carbon Hybrids in Terms of Polymer Electrolyte Membrane Fuel Cell Catalyst Support.

Support materials are of great interest in order to improve the activity and stability of the polymer electrolyte membrane fuel cell (PEMFC) catalysts. Metal oxides have been reported as promising support materials due to their excellent mechanical resistance and high stability against corrosion emerging at acidic and oxidative environment. In this study, high (250 m2/g) and low (45 m2/g) surface area mesoporous TiO2 and high (220 m2/g) and low (30 m2/g) surface area mesoporous Al2O3 were investigated as an alternate cathode catalyst support materials for PEMFCs. These semiconducting TiO2 and Al2O3 metal oxides were combined with the carbon black (Vulcan XC 72) at different mass ratios in order to preserve electrical conductivity of catalyst support a certain extent. Pt and TiO2/C and Pt and Al2O3/C catalysts were prepared by means of Pt reduction on support materials via microwave irradiation technique. The as-prepared catalysts were characterized with some physicochemical and electrochemical analyses. The results reveal that two surface areas TiO2 and Al2O3 support materials differ from each other in terms of fuel cell performance and high surface area TiO2/C (25:75) hybrid supported Pt catalyst gave the best performance.

Both Granulocytic and Non-Granulocytic Blood Cells Are Affected in Patients with Severe Congenital Neutropenia and Their Non-Neutropenic Family Members: An Evaluation of Morphology, Function, and Cell Death

Objective: To examine granulocytic and non-granulocytic cells in children with severe congenital neutropenia (SCN) and their non-neutropenic parents. Materials and Methods: Fifteen patients with SCN and 21 non-neutropenic parents were evaluated for a) CD95, CD95 ligand, annexin V, propidium iodide, cell cycle, and lymphocyte subsets by flow cytometry; b) rapid cell senescence (of leukocytes) by senescence-associated ß-galactosidase stain; c) aggregation tests by aggregometer; d) in vitro bleeding time by PFA-100 instrument; e) mepacrine-labeled dense granule number of thrombocytes by fluorescence microscope; and f) hematomorphology by light and electron microscope. HAX1, ELANE, G6PC3, CSF3R, and JAGN1 mutations associated with SCN were studied in patients and several parents. Results: Significant increase in apoptosis and secondary necrosis in monocytes, lymphocytes, and granulocytes of the patients and parents was detected, irrespective of the mutation type. CD95 and CD95 ligand results implied that apoptosis was non-CD95-mediated. Leukocytes of 25%, 12.5%, and 0% of patients, parents, and controls showed rapid cell senescence. The cell cycle analysis testable in four cases showed G1 arrest and apoptosis in lymphocytes of three. The patients had HAX1 (n=6), ELANE (n=2), G6PC3 (n=2), and unidentified (n=5) mutations. The CD3, CD4, and NK lymphocytes were below normal levels in 16.6%, 8.3%, and 36.4% of the patients and in 0%, 0%, and 15.4% of the parents (controls: 0%, 0%, 5.6%). The thrombocytes aggregated at low rates, dense granule number/thrombocyte ratio was low, and in vitro bleeding time was prolonged in 37.5%-66.6% of patients and 33.3%-63.2% of parents (vs. 0% in controls). Under electron and/or light microscope, the neutrophils, monocytes, lymphocytes, and thrombocytes in the peripheral blood of both patients and parents were dysplastic and the bone marrow of patients revealed increased phagocytic activity, dysmegakaryopoiesis, and necrotic and apoptotic cells. Ultrastructurally, thrombocyte adhesion, aggregation, and release were inadequate. Conclusion: In cases of SCN, patients' pluripotent hematopoietic stem cells and their non-neutropenic parents are both affected irrespective of the genetic defect.

The effect of FV-426 G/A gene variation on the occurrence of thrombosis.

Factor V (FV) plays a crucial role in both procoagulant and anticoagulant pathways. It was indicated that -426G/A change in the promoter of the FV gene may have an effect on the occurrence of thrombosis. We aimed to investigate the effect of -426G/A gene variation on thrombus formation in the thrombosis patients of age 0 and 18 years and 70 years and older. The study included 179 patients with the diagnosis of thromboembolism and also 221 healthy individuals as controls. The Polymerase chain reaction-restriction fragment length polymorphism method was used to detect -426G/A. It was observed that carrying AA genotype does not have a significant risk of thrombosis between the patients and controls (for 0-18 age group P = .96, odds ratio [OR]: 1.13 [0.38-3.30], for ≥70 age P = .94, OR: 0.69 [0.11-4.28]). The same results were obtained when FV Leiden mutation carriers were excluded (for 0-18 age group P = .95, OR: 1.15 [0.35-3.78], for ≥70 age P = .89, OR: 0.73 [0.11-4.56]). In this study, an effect on thrombosis of -426G/A polymorphism was not determined.

Blood group genotyping in multi-transfused patients.

BACKGROUND: In chronically transfused patients, the classical hemagglutination assays may be inaccurate in defining the RBC phenotypes of the patients due to previous transfusions. DESIGN: DNA samples from 39 multi-transfused patients including thalassemia and sickle cell disease were used for red blood cell genotyping. The Rh-Type and KKD-Type (BAGene, BAG Healthcare) were used to determine the polymorphisms associated with antigen expression for RHD, RHCE and Kell, Kidd, Duffy blood group systems, respectively. Results were compared with previously determined phenotyping results for RhD, RhCcEe and Kell by hemagglutination method. RESULTS: Nineteen out of the 37(51%) patients had discrepancies between genotyping and phenotyping results in a total of 25 alleles. In 12 patients, the discrepancies had the potential of alloimmunization. CONCLUSION: Blood group genotyping has vital importance in transfusion management of chronically transfused patients especially if the patients were not phenotyped before starting the initial transfusions.

Determination of factor V Leiden mutation and R2 polymorphism in cis position.

FVA4070G (R2 polymorphism) influences plasma factor V (FV) concentration and was associated with mild activated protein C resistance. This polymorphism was reported to have a trans inheritance with FV Leiden (FVL) mutation. The aim of this study is to investigate the inheritance of R2 polymorphism in the homozygous FVL carriers. In this study, 99 patients with thrombosis and 7 individuals without a history of thrombosis all of which homozygous for FVL were included. Of 99 patients, 1 was heterozygous for FV A4070G. Additionally, 6 polymorphisms in the FV gene were analyzed for the heterozygous R2 patient and her family. When the allelic distribution was classified, 8 different haplotypes were obtained. In contrast to the literature, it was shown that R2 polymorphism could be inherited in cis position with FVL and also the family members could have co-inheritance of the FVL and R2 on the same chromosome as proband.

Intron F G79A polymorphism of the protein Z gene in Turkish Behçet patients.

PURPOSE: Vasculitis is one of the major findings of Behçet's disease (BD). Protein Z (PZ) is a glycoprotein that acts as a cofactor of PZ-dependent protease inhibitor and suppresses trombus formation by inhibiting activated factor Xa. Polymorphism of the PZ gene was mentioned as a genetic risk factor for various thrombotic events. The aim of this study is to investigate the intron F G79A polymorphism of the PZ gene in Behçet patients with and without ocular involvement. METHODS: Seventy-six patients and 70 controls were included in the study. Intron F G79A polymorphism of PZ gene was determined by polymerase chain reaction based DNA analysis. The frequency of A allele and the distribution of genotypes were assessed by χ(2) test and the genotype distribution and Hardy-Weinberg equilibrium were tested with the χ(2) test for quality of fit. RESULTS: The frequency of the A allele was significantly higher in overall Behçet patients than in controls (odds ratio [OR] = 6.8; 95% CI, 2.6 to 17.9; p = 0.0001). It was also significantly higher in patients with (OR = 5.3; 95% CI, 1.83 to 15.6; p = 0.0024) or without (OR = 8.2; 95% CI, 2.95 to 22.5; p = 0.0001) ocular involvement compared to controls. However, A allele frequency was not significantly different between patients with eye involvement versus patients without eye involvement (OR = 0.65; 95% CI, 0.3 to 1.4; p = 0.28). CONCLUSIONS: Although thrombosis in BD is multifactorial, intron F G79A polymorphism of PZ gene in BD may be one of the factors that contribute to this pathological process.

Association between the -402GA, -401GT, and -323ins10-bp polymorphisms of factor VII gene and breast cancer.

BACKGROUND AND PURPOSE: Recent data have shown that the hemostatic system may play a role in cancer development and progression. To test whether factor VII (FVII) can be a candidate factor for breast cancer, we have evaluated the distribution of FVII gene polymorphisms in breast cancer patients and healthy subjects. METHODS: The nested case-control study consisted of 92 women with breast cancer (group 1) and 80 control subjects (in age-matched women) (group 2). Genotyping of the -323ins10-bp, -401GT, and -402GA polymorphisms of the FVII gene was performed by the method of single-strand conformation polymorphism analysis and sequencing. RESULTS: A significant difference was observed in the distribution of the -402GA genotype and allele frequencies in breast cancer and control cases (p < 0.05). For other polymorphisms of the FVII gene, the distributions of genotypes and allele frequencies were not significantly different between two groups (p > 0.05). There was also a significant difference between the distributions of the haplotypes in breast cancer patients and control subjects (p < 0.05). CONCLUSION: Although the number of cases in this study was small, the preliminary findings revealed a possible contribution of the FVII -402GA polymorphism in the development of breast cancer. However, further case-control studies with larger series are needed to confirm our findings.

Association of the tumor necrosis factor-alpha -308 G/A polymorphism with nasal polyposis.

Nasal polyposis (NP) is a chronic inflammatory disease in which several molecular and cellular interactions play important roles. Tumor necrosis factor-alpha (TNF-alpha) is a major pro-inflammatory cytokine with a key role in immune inflammatory responses in NP. Altered levels of TNF-alpha, which may occur due to polymorphisms in the TNF-alpha promoter region, may also be associated with NP susceptibility. Given these facts, we investigated the possible association of the TNF-alpha -308 G/A single nucleotide polymorphism (SNP) with NP. In this study, 97 consecutive adult patients with NP and 95 age- and gender-matched controls were recruited. For identification of SNP, restriction fragment length polymorphism analysis after polymerase chain reaction was carried out. The NP group had a significantly higher rate of polymorphism compared to controls (p = 0.015). Logistic regression analysis revealed that the presence of the TNF-alpha -308 G/A SNP is an independent risk factor for NP development (OR, 3.68; CI, 1.27-10.7; p = 0.016). The presence of a mutation failed to influence disease severity on the basis of resistance to medical and/or surgical treatment. This study suggests a possible linkage of a SNP in the TNF-alpha promoter with NP. These results need to be confirmed with multicentre studies for more precise interpretation and corroborative studies for investigating the influence of polymorphism on transcriptional activity.

Protein Z G79A polymorphism in patients with severe sepsis.

The aim of the study is to investigate whether the presence of a protein Z polymorphism is a risk factor for the development and outcome of sepsis. Sepsis is a clinical syndrome characterized by the presence of systemic signs and symptoms of inflammation. When sepsis leads to organ failure, the term severe sepsis and septic shock is used. The genetic causes of severe sepsis are not fully explained. Protein Z is a vitamin K- dependent glycoprotein and a member of the coagulation cascade. The study included 53 patients with severe sepsis and 70 control healthy volunteers without a familial history of thrombosis. The G79A polymorphism of intron F of the protein Z gene was analyzed by the method of polymerase chain reaction-based DNA analysis. The protein Z intron F G79A polymorphism frequencies of the patients and controls were 43.4% and 40%, respectively. Carrying 79 AA genotype could be a risk factor for severe sepsis and septic shock (OR = 4.5, 95% CI: 0.45-46.1), but it could not find any difference between survivor and nonsurvivor groups. They concluded that the frequency of intron F G79A polymorphism of protein Z gene was higher in patients than controls, and carrying 79 AA genotype could be a risk factor for severe sepsis and septic shock.

Intron F G79A polymorphism of the protein Z gene in cancer patients with and without thrombosis.

BACKGROUND AND PURPOSE: Venous thromboembolism (VTE) is well-recognized complication of cancer; however its pathogenesis is not entirely established. Protein Z (PZ) is a member of the coagulation cascade. We aimed to investigate whether intron F G79A polymorphism of PZ gene is associated with risk of VTE in cancer patients. MATERIALS AND METHODS: The study consisted of 55 cancer patients who developed thrombosis (group 1) and 115 cancer patients without VTE (group 2). Intron F G79A of PZ gene, factor V Leiden (FVL) and prothrombin (PT) G20210A polymorphisms were determined by the method of polymerase chain reaction-based DNA analysis. RESULTS: The prevalence of FVL was significantly greater in group 1 compared with group 2 (27.3% vs. 3.5%, P < 0.0001). No differences were seen in genotype and allele frequencies of PZ gene between two groups (P > 0.05). When the analysis was also made after excluding FVL positive patients, the same insignificant result was found. CONCLUSION: Intron F G79A polymorphism of PZ gene does not contribute to meaningful diagnostic investigation of thrombophilia in cancer patients.

Protein Z G79A polymorphism in Turkish pediatriccerebral infarct patients.

Protein Z (PZ) plays an enhancer role in coagulation as an anticoagulant. This is the first study in which G79A polymorphism investigated in Turkish paediatric stroke patients. Ninety-one paediatric stroke patients with cerebral ischemia and 70 control subjects were analyzed for PZ G79A and also FVL, PT mutations. PZ 79 'A' allele in homozygous state was found in five patients (5,5%), while it was found only in one control subject (1,4%) and it was seemed as a risk factor for peadiatric ischemia [OR=3,94 (0,44-35,1)]. When patients and controls who had FVL and PT carriers were excluded, AA genotype carried a risk [OR=3,88 (0,41-36,5)]. Also plasma protein Z levels measured in 21 stroke patients and 52 controls. Plasma protein Z levels were not different between stroke patients (500,95 ngmL-1±158,35) and controls (447,34 ngmL-1±165,97). But the plasma levels of protein Z was decreased in patients with AA genotype. Our data showed that carrying 79 AA genotype could be a genetic risk factor for cerebral infarct in peadiatric patients.

Prevalence of the MEFV gene mutations in childhood polyarteritis nodosa.

OBJECTIVES: To test the hypothesis that alterations in the Mediterranean fever (MEFV) gene are a susceptibility factor for the development of polyarteritis nodosa (PAN) we investigated the prevalence of MEFV mutations in patients with PAN without any symptoms of familial Mediterranean fever (FMF). STUDY DESIGN: Pediatric patients with PAN (n = 29) were enrolled in this study. Six predominant mutations (p.M694V, p.M680I, p.M694I, p.V726A, p.K695R, p.E148Q) in the MEFV gene were studied. RESULTS: Fifteen MEFV mutations were identified in 58 chromosomes. Eleven of the 29 patients (38%) were found to carry MEFV mutations. Three (10.3%) of them had homozygous p.M694V mutation, and one of the patients (3.4%) had compound heterozygous mutation (p.V726A/p.E148Q). CONCLUSIONS: Our study confirms that alterations in the MEFV gene are important susceptibility factors for the development of PAN. We believe that mutations in MEFV gene provide a basis for the development of PAN both by forming a proinflammatory state and by possibly giving exaggerated response to streptococcal infections.

Serum amyloid A1 -13 T/C alleles in Turkish familial Mediterranean fever patients with and without amyloidosis.

Previously reported studies concerning the effect of homozygosity of the 1.1 allele of the SAA gene found a correlation between this haplotype and susceptibility to amyloidosis in FMF patients. Another report revealed a strong association between SAA1 -13T/C and secondary amyloidosis in the rheumatoid arthritis patient group. In this study, we aimed to determine the effect of SAA1 -13T/C in FMF patients with and without amyloidosis. The study cohort, consisting of 166 patients with FMF was divided into two groups, according to the presence (n=66) or absence (n=100) of renal amyloidosis at study entry. MEFV gene mutation analysis and allelic variant of SAA1 gene -13 T/C was analyzed according to the previously described techniques. SAA1 -13 T allele frequencies were 0.5816, 0.23 and 0.4242 in controls, FMF patients and FMF-amyloidosis patients respectively. The difference between controls vs. FMF patients and FMF-amyloidosis patients were 0.0002 and 0.1673 respectively. It was 0.0071 for FMF-patients vs. FMF-amyloidosis. When 694 M/V homozygous nonamyloid-FMF group was compared with 694 M/V carriers of the FMF-amyloidosis group, the difference was 0.049. When carrying TT allele was considered, the difference between controls vs. FMF patients and FMF-amyloidosis patients were 0.0001 and 0.58. It was 0.0003 for FMF patients vs. FMF-amyloidosis. When 694 M/V homozygous nonamyloid-FMF group was compared with 694 M/V carriers of the FMF-amyloidosis group, the difference was 0.03. Carrying SAA -13T in homozygote state revealed a 7.9 (95% CI 3.6 -17.5) fold risk for the occurrence of amyloidosis when compared with FMF patients without amyloidosis. This was 8.75 (95% CI 3.0 - 25.1) when 694 M/V homozygotes were taken into consideration. Our data revealed that the genotype SAA1 -13T has at least an effect on the development of amyloidosis. As more data on this polymorphism accumulate, we will understand its effect on the pathogenesis of amyloidosis in FMF.

Interleukin-6 (IL-6) -174 G/C polymorphism in familial Mediterranean fever patients with and without amyloidosis.

Familial Mediterranean fever (FMF) is a recessive disorder characterized by attacks of fever and inflammation. A sustained inflammatory reaction is observed in the disease course, and cytokine levels such as interleukin (IL)-1, IL-6 and tumor necrosis factor-alpha (TNF-alfa) are shown to increase during and between the attacks. In this study, we investigated the role of the functionally important IL-6 -174 G/C polymorphism in the clinical outcome of FMF and amyloidosis. One hundred and fifty-six FMF patients (80 with amyloidosis) and 90 healthy controls were studied. The genotype distributions and allele frequencies of the patients and the controls were found to be similar, and the differences between the groups were not statistically significant. The results show that IL-6 -174 G/C polymorphism is not associated with FMF and amyloidosis. The increase observed in cytokine levels during and between the attacks is more likely due to the inflammatory nature of the disease.