A combined allele-specific PCR and RFLP assay to detect the 35delG mutation in the Connexin 26 gene

Mutations in the Connexin-26 (specified GJB2) gene have been shown to be a major cause of nonsyndromic recessive deafness (NSRD), and a single mutation 35delG in the GJB2 gene accounts for the majority of cases of NSRD. For diagnostic analyses and for scientific studies of large numbers of patients, fast and economic assays that can be performed with standard polymerase chain reaction (PCR) instruments are highly desirable. We have developed an allele-specific amplification (ASA)-based restriction fragment length polymorphism (RFLP) assay. We evaluated the multiplex method for its ability to 35delG mutation. Our method is a stable, reproducible and concordend with previously reported PCR-RFLP assays.     

A research on the microfungal flora of somegreenhouse soils in the vicinity of LapsekiÇanakkale,Turkey

Qualitative and quantitative distributions of microfungal flora of the some greenhouse soils in the vicinity of Lapseki, Canakkale were studied. A total of 25 soil samples were collected from 10 cm depth in five greenhouses. Using soil dilution technique, 128 different microfungal isolates were obtained Isolates collected belonged to Oomycetes (1), Zygomycetes (7), Ascomycetes (9) and 96 belonging to the Deuteromycetes Fifteen isolates were classified as Mycelia Sterilia. The genera of microfungi most abundant in the greenhouse soils were Aspergillus, Penicillium, Geomyces, Exophiala and Fusarium. Qualitative and quantitative distributions of the Aspergillus were higher than those of the other genera. It was found that the maximum value was in greenhouse 5 and the minimum value was in greenhouse 4. The solarization method was the most effective in reducing fungal numbers.     

Effects of chronic fluorosis on electrocardiogram in sheep

This study was carried out to evaluate the effects of chronic fluorosis by means of the electrocardiograms in sheep. Ten sheep with fluorosis living around a volcanic mountain (Tendürek Mount) in East Anatolia in Turkey and 10 healthy sheep were used. Leads I, II, III, aVR, aVL, aVF, V₂, V₄, and V₁₀ were recorded in the electrocardiographs of the sheep. All waves were seen in all derivations. The P-Q interval was significantly (p<0.05) prolonged and sinus bradycardia was observed in the sheep with fluorosis. As a result of this, the number of heart beats was decreased significantly (p<0.05); that is, the number of heart beats was 110±15 in the control group and 75±10 in sheep with fluorosis.     

Analyses of functional IL10 and TNF-α genotypes in Behçet’s syndrome

We aim to ascertain the possible involvement of functional IL10 and TNF-α promoter polymorphisms on the susceptibility to Behçet’s syndrome (BS), to examine whether IL10 and TNF-α genotypes might work synergistically influencing susceptibility to BS. IL10 ?1082G/A, ?819C/T and ?592C/A and TNF ?308G/A polymorphisms were analyzed in 102 Turkish patients with BS and 102 healthy subjects by using amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). We have found no significant associations between IL10 ?1082G/A, ?819C/T, ?592C/A, TNF-α ?308G/A polymorphisms and BS. Also, no significant correlation was found between IL10 GCC, ACC, ATA haplotypes, GCC⁺/GCC⁺, GCC⁺/GCC^?, GCC^?/GCC^? genotypes. There was no significant association between combined TNF-α/IL10 genotypes and BS. Our study indicates that functional TNF-α, IL10 genotypes or combined TNF-α, IL10 genotypes do not play a role in BS susceptibility in Turkish BS patients.     

Effects of Calcium Supplementation on Glucose and Insulin Levels of Athletes at Rest and After Exercise

This study was performed to determine how the calcium supplementation for a 4-week period affects the glucose and insulin levels at rest and at exhaustion in athletes. This is a 4-week study performed on 30 healthy subjects varying between 18 and 22 ages. Subjects were separated into three groups: first group (group supplemented with calcium, sedentary group), second group (calcium supplementations + exercise group), and third group (training group). Glucose and insulin parameters of the groups were measured four times, at rest and exhaustion in the beginning of the research and at rest and exhaustion after the end of 4 weeks application period. Exhaustion measurements both before and after the supplementations significantly decreased in compared to rest measurements in terms of insulin (p < 0.05). Significant difference was not determined in the glucose values of groups. In terms of glucose, values increased in all of the three groups occurred with exercise both before and after the supplementation by exercise and exhaustion (p < 0.05). The results of our study indicate that calcium gluconate supplementations for 4 weeks in sedentary subjects and athletes did not significantly affect plasma insulin levels at rest and exhaustion. However, glucose levels were affected by calcium supplementation and exhausting exercise in athletes.     

The effects of magnesium supplementation on thyroid hormones of sedentars and Tae-Kwon-Do sportsperson at resting and exhaustion

The effect of magnesium on thyroid hormones of sedentars and sportsperson in Tae-Kwon-Do, has been investigated in a 4-weeks training program. Group 1 consisted of sedentars receiving 10 mg/kg/day Mg for 4 weeks. Group 2 consisted of subjects receiving magnesium (Mg) supplement and practicing Tae-Kwon-Do for 90-120 min/day, for five days a week. Group 3 consisted of subjects practicing Tae-Kwon-Do but receiving Mg supplements. TSH levels increased with training and Mg supplementation (p<0.05). Mg increased FT3 values. (p<0.05). TT3 values of groups reduced in all groups (p<005). After supplementation, group 1 had higher TT4 values than groups 1 and 3 and the group 2 had higher TT4 values than the third group (p<005). Results of this research show that training until exhaustion causes reduction in thyroid hormone activity in sedentars and sportsperson. It has been established that Mg supplementation however, prevents reduction in thyroid hormone activity in sedentars and sportsperson.     

Aquaporin-1 channel function is positively regulated by protein kinase C

Aquaporin-1 (AQP1) channels contribute to osmotically induced water transport in several organs including the kidney and serosal membranes such as the peritoneum and the pleura. In addition, AQP1 channels have been shown to conduct cationic currents upon stimulation by cyclic nucleotides. To date, the short term regulation of AQP1 function by other major intracellular signaling pathways has not been studied. In the present study, we therefore investigated the regulation of AQP1 by protein kinase C. AQP1 wild type channels were expressed in Xenopus oocytes. Water permeability was assessed by hypotonic challenges. Activation of protein kinase C (PKC) by 1-oleoyl-2-acetyl-sn-glycerol (OAG) induced a marked increase of AQP1-dependent water permeability. This regulation was abolished in mutated AQP1 channels lacking both consensus PKC phosphorylation sites Thr(157) and Thr(239) (termed AQP1 DeltaPKC). AQP1 cationic currents measured with double-electrode voltage clamp were markedly increased after pharmacological activation of PKC by either OAG or phorbol 12-myristate 13-acetate. Deletion of either Thr(157) or Thr(239) caused a marked attenuation of PKC-dependent current increases, and deletion of both phosphorylation sites in AQP1 DeltaPKC channels abolished the effect. In vitro phosphorylation studies with synthesized peptides corresponding to amino acids 154-168 and 236-250 revealed that both Thr(157) and Thr(239) are phosphorylated by PKC. Upon stimulation by cyclic nucleotides, AQP1 wild type currents exhibited a strong activation. This regulation was not affected after deletion of PKC phosphorylation sites in AQP1 DeltaPKC channels. In conclusion, this is the first study to show that PKC positively regulates both water permeability and ionic conductance of AQP1 channels. This new pathway of AQP1 regulation is independent of the previously described cyclic nucleotide pathway and may contribute to the PKC stimulation of AQP1-modulated processes such as endothelial permeability, angiogenesis, and urine concentration.     

Nanotube action between human mesothelial cells reveals novel aspects of inflammatory responses

A well-known role of human peritoneal mesothelial cells (HPMCs), the resident cells of the peritoneal cavity, is the generation of an immune response during peritonitis by activation of T-cells via antigen presentation. Recent findings have shown that intercellular nanotubes (NTs) mediate functional connectivity between various cell types including immune cells - such as T-cells, natural killer (NK) cells or macrophages - by facilitating a spectrum of long range cell-cell interactions. Although of medical interest, the relevance of NT-related findings for human medical conditions and treatment, e.g. in relation to inflammatory processes, remains elusive, particularly due to a lack of appropriate in vivo data. Here, we show for the first time that primary cultures of patient derived HPMCs are functionally connected via membranous nanotubes. NT formation appears to be actin cytoskeleton dependent, mediated by the action of filopodia. Importantly, significant variances in NT numbers between different donors as a consequence of pathophysiological alterations were observable. Furthermore, we show that TNF-α induces nanotube formation and demonstrate a strong correlation of NT connectivity in accordance with the cellular cholesterol level and distribution, pointing to a complex involvement of NTs in inflammatory processes with potential impact for clinical treatment.