Evaluation of ciliary functions and ciliary beat frequency via cell culture method in patients with primary ciliary dyskinesia.

BACKGROUND: Cell culture increases both diagnostic specificity and sensitivity of primary ciliary dyskinesia (PCD) and the most important reason to use cell culture for definitive diagnosis in PCD is to exclude secondary ciliary defects. Here we aimed to evaluate the cilia functions and cilia ultrastructural abnormalities after ciliogenesis of cell culture in patients with definitive diagnosis of PCD. We also aimed to compare high speed videomicroscopy (HSVM) results of patients before and after ciliogenesis and to compare them with electron microscopy, genetic and immunofluorescence results in patients with positive diagnosis of PCD. METHODS: This study was conducted as a cross-sectional study in patients with PCD. HSVM, transmission electron microscopy (TEM) and immunofluorescence staining results of the nasal biopsy samples taken from patients with the definitive diagnosis of PCD were evaluated and HSVM findings before and after cell culture were described. RESULTS: Ciliogenesis and regrowth in the cell culture occurred in the nasal biopsy sample of eight patients with PCD. The mean age of the patients was 15.5±4.2 years (8.5-18 years). Mean beat frequency was found to be 7.54±1.01 hz (6.53-9.45 hz) before cell culture, and 7.36±0.86 hz (6.02-7.99 hz) after cell culture in the nasal biopsy of patients. There was no significant difference in the beat frequency of PCD patients before and after cell culture. Ciliary function analysis showed the similar beating pattern before and after cell culture in patients with PCD. CONCLUSIONS: This study showed us that there was no difference between cilia beat frequency and beat pattern before and after cell culture in patients with definitive diagnosis of PCD and repeated HSVM would be a useful diagnostic approach in patients who have no possibility to reach other diagnostic methods.

Evaluation of umbilical cord immune cells in pregnancies with autoimmune disorders and/or methylenetetrahydrofolate reductase polymorphisms.

OBJECTIVES: To evaluate umbilical cord immune cells in pregnancies with autoimmune disorders (AID) and/or methylenetetrahydrofolate reductase (MTHFR) polymorphisms. METHODS: Umbilical cords were obtained from seven AID women without MTHFR polymorphisms, eight with AID and MTHFR polymorphisms, nine with MTHFR polymorphisms, and eight with neither. Umbilical cords were assessed immunohistologcally by anti-CD4, anti-CD8, anti-CD14, anti-CD19, anti-CD21, and anti-CD56 antibodies in six umbilical cord zones: 1) arterial wall 2) periarterial zone 3) venous wall 4) perivenous zone 5) intervascular zone, and 6) subamniotic zone. RESULTS: AIDs and MTHFR polymorphisms had an effect on the number and composition of CD4+ cells in the venous wall. The presence of a MTHFR polymorphism may affect the number and morphology of CD4+ cells in the subamniotic zone. CD8+ cell distribution is substantially influenced by the presence of maternal risk factors. The co-existence of AID with MTHFR polymorphism has a prominent effect on the number and morphology of CD14+ cells, especially in the arterial wall. CD19+ cells were only observed in the control group in the venous wall, perivenous zone, and intervascular zone. CD21+ cells were only observed in the arterial wall of the control group and the intervascular zone of the AID group with different morphologic features. The number and morphology of CD56+ cells is prominently affected by the presence of maternal risk factors. CONCLUSIONS: Umbilical cord stem cell and immune cell composition may be affected by the presence of risk factors like MTHFR polymorphisms and/or AID.

A low direct electrical signal attenuates oxidative stress and inflammation in septic rats.

Electrical stimulation is proposed to exert an antimicrobial effect according to studies performed using bacterial and cell cultures. Therefore, we investigated the effects of electrification on inflammation in septic rats. Twenty-eight male Wistar albino rats were divided into 4 groups: healthy control (C), electrified healthy (E), sepsis (S), and electrified sepsis (SE) groups. Staphylococcus aureus (1 x 109 colonies) in 1 ml of medium was intraperitoneally injected into rats to produce a sepsis model. The rats in the E and SE groups were exposed to a low direct electrical signal (300 Hz and 2.5 volts) for 40 min and 1 and 6 h after bacterial infection. Immediately after the second electrical signal application, blood and tissue samples of the heart, lung, and liver were collected. An antibacterial effect of a low direct electrical signal was observed in the blood of rats. The effects of electrical signals on ameliorating changes in the histological structure of tissues, blood pH, gases, viscosity and cell count, activities of some important enzymes, oxidative stress parameters, inflammation and tissue apoptosis were observed in the SE group compared to the S group. Low direct electrical signal application exerts antibacterial, antioxidant, anti-inflammatory and antiapoptotic effects on septic rats due to the induction of electrolysis in body fluids without producing any tissue damage.

Alterations of Tear Film and Ocular Surface in Children with Type 1 Diabetes Mellitus.

Purpose: To investigate whether diabetes mellitus (DM) affects ocular surface of children with well-controlled type 1 DM.Methods: Sixty-five diabetic patients and 55 age-matched controls enrolled to study. Detailed ocular surface assessment including, ocular surface disease index (OSDI) questionnaire, tear film break-up time (TBUT) analysis, Schirmer test, and conjunctival impression cytologic analysis were performed.Results: Schirmer test and TBUT results were significantly lower in DM group than controls (p = 0.001, for all). OSDI scores of all participants were within normal range. Impression cytology analysis showed grade 0 changes in all participants and there was no difference between groups for goblet cell density (p > 0.05). The TBUT results were significantly associated with duration of DM (r = -0.309, p = 0.036).Conclusion: Diabetic children without symptoms, signs, and definite diagnosis of dry eye still had lower TBUT and Schirmer test results than controls; however, impression cytology analysis was similar in both groups.

Local Ocular Surface Alterations in Children with Hashimoto's Thyroiditis.

PURPOSE: To evaluate the ocular surface characteristics based on Schirmer's test, tear break-up time (TBUT), and conjunctival impression cytology (CIC) in children with Hashimoto's thyroiditis (HT). METHODS: This study included 51 children with HT and 53 control subjects. The ocular surface characteristics of participants were assessed via Schirmer's test, TBUT, and CIC. Conjunctival samples were examined cytologically according to the Nelson grading system. RESULTS: Schirmer's and TBUT results were significantly lower in HT group (p < .05). All samples in both the study and control groups were evaluated as grade 0 according to the Nelson classification (p = .841), however, goblet cell density (GCD) was significantly lower in HT group (p = .001). Schirmer test results were significantly associated with the duration of HT (p = .025, r = -0.311). CONCLUSION: Hashimoto's thyroiditis without any ocular complaints may cause ocular surface changes with TBUT and Schirmer's. Although CIC analysis showed similar grading results, GCD was significantly decreased in HT group.

Angiotensin-converting enzyme inhibitor enalapril reduces formation of hypertrophic scars in a rabbit ear wounding model.

BACKGROUND: Angiotensin-converting enzyme inhibitors are widely used in medicine because of their antihypertensive and antifibrogenic effects. Angiotensin-converting enzyme activates angiotensin I to angiotensin II, which plays an important regulatory role in wound healing and collagen production. The authors investigated whether systemic administration of angiotensin-converting enzyme inhibitors has any effect on formation of hypertrophic scars using the rabbit ear wound model. METHODS: Sixteen New Zealand albino rabbits were divided into four groups, and four punch defects were created on each ear. The first group received oral enalapril immediately after the creation of punch defects. The second group received oral enalapril on day 28 after the formation of scars. The third group received intralesional steroid injections on days 28 and 35. The fourth group was the control group. The rabbits were killed on day 40. The harvested specimens were analyzed histomorphometrically and immunohistochemically. RESULTS: Early enalapril application decreased the scar elevation index and fibroblast and capillary counts significantly, compared with the values in the control group. Late enalapril application decreased fibroblast counts significantly; however, there was no difference in scar elevation index compared with the control group. There was no difference between early enalapril application and steroid therapy in terms of scar elevation index and capillary and fibroblast counts. However, early and late enalapril groups displayed lower collagen type III immunoreactivity compared with the steroid and control groups. CONCLUSION: Early application of enalapril following dermal injury reduces formation of hypertrophic scars, probably because of its down-regulatory effects on type III collagen production.

Prefabricated osteocutaneous neural island flap model.

Neural-based flaps are an interesting clinical choice particularly in difficult cases that may not be reconstructed with known techniques. Their popularity is gradually increasing because these flaps offer the advantage of preservation of major extremity arteries and avoidance of microsurgical techniques. Our aim was to explore the feasibility of prefabrication of an osteocutaneous neural island flap model in this study. A peripheral nerve of the rat was implanted into the subcutaneous tissue of a skin flap that was connected to a segment of bone by a soft-tissue bridge, to prefabricate an osteocutaneous flap that was supplied only by the intrinsic vasculature of that nerve after a preliminary delay period. At the end of this study, based on direct observation, microangiographic findings, and additionally, a detailed histologic analysis consisting of both qualitative and quantitative assessments, we have proved that it was possible to prefabricate an osteocutaneous composite flap based on the vascularity of a peripheral nerve after a 2-step delay period. We believe that the clinical application of this new flap will gradually develop based on further experimental studies.