Walter Krugluger

Increased visfatin concentrations in women with gestational diabetes mellitus

The recently discovered adipocytokine visfatin has insulin-like properties. It lowers blood glucose and improves insulin sensitivity; however, clinical data on visfatin are limited. To evaluate the role of visfatin in GDM (gestational diabetes mellitus), we determined visfatin levels in women with GDM and in healthy pregnant controls. Furthermore, visfatin concentrations were investigated longitudinally during pregnancy and after delivery in a subgroup of women with GDM. Blood for measurement of visfatin and metabolic parameters was obtained from 64 women with GDM [median week of gestation, 34 (interquartile range, 27-36) weeks] and 30 healthy pregnant controls [median week of gestation, 34 (interquartile range, 28-36) weeks]. In a subgroup of 24 women with GDM, visfatin, leptin and metabolic parameters were investigated twice during pregnancy (28-30 and 38-40 weeks of gestation) and 2 weeks after delivery. In the cross-sectional analysis, median visfatin levels were significantly elevated in women with GDM [64.0 (interquartile range, 50.9-74.8) ng/ml] compared with controls [46.0 (interquartile range, 36.9-54.6) ng/ml; P<0.0001]. In women with GDM, visfatin correlated with week of gestation at the time of blood draw (R=0.35, P=0.005). No association with fasting glucose, insulin, homoeostasis model assessment-insulin resistance or body mass index was observed. According to the longitudinal analysis, visfatin increased during pregnancy (P=0.002) and rose further after delivery (P=0.014), whereas leptin and insulin levels decreased after parturition (both P<0.001). In conclusion, visfatin is elevated in women with GDM and increases during the course of pregnancy as well as after delivery. Furthermore, visfatin shows no association with insulin and leptin in women with GDM.

Plasminogen Activator Inhibitor 1 4G/5G Polymorphism and Coagulation Factor XIII Val34Leu Polymorphism: Impaired Fibrinolysis and Early Pregnancy Loss

BACKGROUND A successful outcome of pregnancy depends on proper placental formation. In the very beginning of this process, trophoblast invasion and fibrin deposition into the wall of the decidual veins play an important part. Two polymorphisms, coagulation factor XIII (FXIII) Val34Leu and plasminogen activator inhibitor 1 (PAI-1) 4G/5G, interfere with fibrin cross-linking and regulation of fibrinolysis and may therefore contribute to early pregnancy loss. METHODS We enrolled 49 unrelated Caucasian women with a history of two consecutive or three to six nonconsecutive early pregnancy losses and 48 unrelated parous healthy controls without a history of pregnancy loss and evaluated them for the following genetic variants: the factor V Leiden and prothrombin G20210A gene mutations, the methylenetetrahydrofolate reductase C677T and A1298C polymorphisms, and the PAI-1 4G/5G and FXIII Val34Leu polymorphisms. RESULTS For the isolated occurrence of PAI-1 4G/5G or FXIII Val34Leu, we found no statistically significant difference between cases and controls. For homozygosity of either or compound carrier status of both mutations, the overall relative risk for early pregnancy loss was significantly increased (odds ratio = 2.4; 95% confidence interval, 1.1-5.5; P = 0.032). We observed no statistically relevant association of any of the other tested mutations with early pregnancy loss. CONCLUSION Homozygosity for PAI-1 4G or FXIII 34Leu polymorphisms as well as compound carrier status is associated with early pregnancy loss.

Increase in Visfatin after Weight Loss Induced by Gastroplastic Surgery

Objective: The recently described adipokine visfatin is produced in visceral fat and has been suggested to influence insulin resistance. To investigate whether visfatin concentrations are related to changes in body weight, this adipokine was measured in insulin‐resistant severely obese patients before and after gastroplastic surgery.

Elevated coagulation factor VIII and the risk for recurrent early pregnancy loss

Inherited and acquired thrombophilia are associated with recurrent pregnancy loss. Recently, an increased risk for thromboembolic disease was described for patients with elevated coagulation factor VIII, but it is unknown whether there is also an association to early pregnancy loss. We therefore evaluated the relation between recurrent early pregnancy loss and levels of coagulation factor VIII. We enrolled 49 unrelated Caucasian women with a history of 2-6 early pregnancy losses and 48 healthy controls, who had delivered at least one term infant and had never experienced pregnancy loss. We determined factor V Leiden-, G20210A prothrombin-, MTHFR C677T- and A1298C-gene mutations, levels of antithrombin, protein C, protein S, factor VIII, C-reactive protein and antiphospholipid antibodies. There was a significantly higher rate of pregnancy losses in women with Antiphospholipid Syndrome (p = 0.043). Furthermore, plasma levels of coagulation factor VIII were significantly higher in cases than in controls (130.5 IU/dl +/- 25.4 vs 119.5 IU/dl +/- 24.1; p = 0.032) and appeared independent of C-reactive protein (R = 0.146, p = 0.323 in cases; R = -0.028, p = 0.850 in controls). The relative risk for recurrent pregnancy loss in women with factor VIII levels above 151 IU/dl (90(th) percentile of controls) was 2.5 (0.7 - 8.9, 95 percent confidence interval), for levels above 156 IU/dl (95(th) percentile of controls) 3.9 (0.8 - 20.0, 95 percent confidence interval). Elevated maternal plasma levels of coagulation factor VIII tend to be associated with an increased risk for recurrent early pregnancy loss.

Validation of a reverse-hybridization StripAssay for the simultaneous analysis of common α-thalassemia point mutations and deletions

Abstract Background: α-Thalassemia is a worldwide disease and considered to be a major public health problem in countries within the so-called thalassemia belt. The complex genetics of α-thalassemias requires diagnostic methods with the capacity to screen rapidly and accurately for common causative mutations. Methods: We developed and validated a reverse-hybridization assay (Alpha-Globin StripAssay) for the rapid and simultaneous detection of 21 α-globin mutations: two single gene deletions (–α3.7; –α4.2), five double gene deletions [--MED; --SEA; --THAI; --FIL; –(α)20.5], αααanti-3.7 gene triplication, two point mutations in the α1 gene (cd 14 G>A; Hb Adana) and 11 point mutations in the α2 gene (initiation cd T>C; cd 19 –G; IVS1 –5nt; cd 59 G>A; Hb Quong Sze; Hb Constant Spring; Hb Icaria; Hb Pakse; Hb Koya Dora; polyA-1; polyA-2). Results: Reliable genotyping of recombinant mutant clones and reference DNA samples was achieved by means of two corresponding test strips presenting parallel arrays of allele-specific oligonucleotides. The entire procedure from blood sampling to the identification of mutations required less than 6 h, and hybridization/detection was manual or automated. The diagnostic potential of this Alpha-Globin StripAssay was carefully evaluated on 272 pre-typed samples in a multicenter validation study. In 96.14% of the cases, StripAssay typing was completely concordant with the reference methods. Conclusions: The Alpha-Globin StripAssay proved to be a fast, easy-to-perform and reliable screening method to identify >90% of α-globin mutations in endemic areas worldwide. Clin Chem Lab Med 2007;45:605–10.

High Prevalence of the −α3.7 Deletion Among Thalassemia Patients in Iran

a-Thalassemia (thal) is one of the most common inherited hemoglobin (Hb) disorders worldwide. It is caused by deletional or nondeletional mutations of one (aþ) or both (a) genes in cis at the a-globin gene cluster on chromosome 16p13.3. Unlike the prevalence of point mutations in b-thal, the majority of a-thal alleles are derived from single gene deletions (e.g. a or a), or from double gene deletions, such as the Southeast Asian ( ) and the Mediterranean ( ) variants (1). In Iran, due to the lack of population data and molecular diagnostic services for a-thal, a considerable number of patients with microcytic, hypochromic anemia, and normal Hb A2 levels are at risk of being misdiagnosed as silent b-thal carriers, or have to be left without molecular diagnosis of the disease. In this study we have tested 57 Iranian individuals from a variety of ethnic origins, randomly chosen from a pool of patients with MCV values below 80.0 fL (mean: 67.4 5.9), low MCH (21.4 2.5 pg), normal or slightly reduced PCV (0.404 0.041 L=L) and Hb (12.6 1.4 g=dL) levels, normal Hb A2 (3.0 1.1%), and negative results in b-thal genotyping, for the two most common a-thal single gene deletions ( a and a). Iron deficiency was ruled out by testing serum iron and ferritin levels, as well as total iron binding capacity (TIBC), which were in the normal range for all these

Activation of the β-Catenin Signaling Pathway and Its Impact on RPE Cell Cycle

PURPOSE To investigate the effect of EGF, IGF-1, and VEGF on ARPE19 cell proliferation and differentiation. METHODS The gene expression of RPE-specific differentiation and proliferation markers and the transcriptional and translational activity of beta-catenin signaling markers were measured by flow cytometry and RT-PCR. RESULTS The data showed a significant decrease in RPE65, CRALBP, and cytokeratin 18 in ARPE-19 cells stimulated with EGF and IGF-1. In addition, a significant decrease in GSK-3beta and beta-catenin was observed that was paralleled by an increase in cyclin D1 expression. Cell cycle studies revealed an increase in ARPE cells in the S-G(2)/M-phase after treatment with EGF or IGF-1. VEGF, on the other hand, led to a reduction in cyclin D1 and to an increase in GSK 3beta and beta-catenin expression which was paralleled by an increase in RPE-specific differentiation markers. CONCLUSIONS The data demonstrate the induction of proliferation by EGF and IGF-1 and upregulation of the beta-catenin signaling pathway in ARPE-19 cells. The data suggest that activation of the beta-catenin signaling pathway may be key in activating ARPE-19 cells by different growth factors.

Pseudotumor cerebri from sinus venous thrombosis, associated with polycystic ovary syndrome and hereditary hypercoagulability

Objective. The association of pseudotumor cerebri, visual impairment, hypothyroidism, polycystic ovary syndrome (PCOS), and a hypercoagulable state due to a factor V and a prothrombin mutation has not been reported previously. Case report. A 20-year-old obese woman developed menstrual cycle irregularities since age 14 years, initially bitemporal and latter diffuse headache since age 14 years, and bilateral visual impairment, described as sparkling black points. Ophthalmologically there was a recurrent papilledema. Clinical neurologic investigations revealed sore neck muscles and hirsutism. Magnetic resonance imaging of the brain, orbita and cervical spine, and investigations of cerebrospinal fluid were non-informative. Visually evoked potentials revealed demyelination of the optic nerves. Gynecologic investigations revealed PCOS and endocrinologic investigations hypothyroidism and hyperandrogenism. Tests for thrombophilia disclosed a heterozygote state for the G1.697A factor V Leiden and the G20.210A prothrombin mutation. A possible relationship between pseudotumor cerebri and the ophthalmologic, gynecologic, endocrinologic and coagulation abnormalities is discussed. Conclusions. For the first time we describe the association of pseudotumor cerebri, optic nerve demyelination, PCOS, other endocrinologic abnormalities, and thrombophilia due to a factor V and prothrombin mutation. A causal relationship between these abnormalities remains elusive.

Implication of CD21, CD35, and CD55 in the pathogenesis of age-related macular degeneration.

PURPOSE To determine a possible implication of CD21, CD35, and CD55 in the pathogenesis of age-related macular degeneration (AMD) by assessing the difference in expression rates of these factors on AMD patients and a control group. DESIGN Case-control study. METHODS Fifty unrelated AMD patients and 48 unrelated sex- and age-matched control subjects participated in this case-control study. Samples of fresh EDTA-blood were stained and flow cytometry was chosen to measure fluorescence emissions. The association between exudative AMD and CD21, CD35, and CD55 was evaluated from all patients who completed the study. RESULTS Our study shows CD35 to be expressed in a significantly higher frequency in AMD patients on monocytes (P = .00586), lymphocytes (P = .000605), and granulocytes (P < .000033). In contrast, the expression rate of CD21 (P > .05) and CD55 (P > .05) are similar in both groups. CONCLUSION More regulative factors of the complement system are involved in pathogenesis of AMD. Our study underlines the key role of the complement system in AMD and shows the involvement of the whole immune system through more regulative factors.

Positive troponin-T in noncompaction is associated with neuromuscular disorders and poor outcome

SummaryObjectivesThough cardiac troponin-T may be positive in hypertrophic and dilative cardiomyopathy, it is not known how often troponin-T is positive in left ventricular hypertrabeculation/ noncompaction (LVHT), an unclassified cardiomyopathy. This retrospective study aimed to assess how often troponin-T is positive in LVHT, is associated with elevated CK, is attributable to cardiac or extra-cardiac causes, in particular neuromuscular disorders (NMDs), or if it is a predictor of poor survival.ResultsAmong 100 patients, detected over a period of 11 years, troponin-T was determined at least once in 71 (71%) of them. Troponin-T was determined once in 36 patients, twice in 8 cases, three times in 11 patients, and more than three times in 16 cases. Troponin- T was positive at least once in 12 patients (17%). Forty-five of the 71 patients suffered from a NMD (63%). Troponin-T positivity was associated with elevated CK in 6 cases. Troponin-T-positivity was attributable to acute myocardial ischemia in a single case, to chronic renal failure in 5 cases, to dilative cardiomyopathy in 4 cases, to atrial fibrillation in 3 cases, to heart failure in 4 cases, and to NMD in 10 cases. Troponin-T positivity in LVHT patients with NMD was assumed to be due to cardiac involvement in the disease. Among the 22 patients who died during the observational period troponin was determined in 16 and was positive in 4 (25%).ConclusionsTroponin-T is positive in 17% of the patients with LVHT. Most of these patients suffer from a NMD. Troponin-T positivity in LVHT predicts the presence of NMD and poor survival.