Comparison of the efficiencies of intrathecal and intraganglionic injections in mouse dorsal root ganglion

Background/aim: Dorsal root ganglia (DRG) are structures containing primary sensory neurons. Intraganglionic (IG) and intrathecal (IT) applications are the most common methods used for viral vector transfer to DRG. We aim to compare the efficiencies and pathological effects of IT and IG viral vector delivery methods to DRG, through in vivo imaging. Materials and methods: Mice were divided into four groups of six each: IT, IG, IT-vehicle, and IG-vehicle. Adeno-associated virus (AAV) injection was performed for EGFP expression in IT/IG groups. DRGs were made visible through vertebral window surgery and visualized with multiphoton microscopy. After imaging, spinal cords and DRGs were removed and cleared, then imaged with light sheet microscopy. Results: No neuronal death was observed after IT injection, while the death rate was 17% 24 h after IG injection. EGFP expression efficiencies were 90%–95% of neurons in both groups. EGFP expression was only observed in targeted L2 DRG after IG injection, while it was observed in DRGs located between L1-L5 levels after IT injection. Conclusion: IT injection is a more suitable method for labeling DRG neurons in neurodegenerative injury models. However, when the innervation of DRG needs to be specifically studied, IT injection reduces this specificity due to its spread. In these studies, IG injection is the most suitable method for labeling single DRG neurons

Active shrinkage protects neurons following axonal transection

Trauma, vascular events, or neurodegenerative processes can lead to axonal injury and eventual transection (axotomy). Neurons can survive axotomy, yet the underlying mechanisms are not fully understood. Excessive water entry into injured neurons poses a particular risk due to swelling and subsequent death. Using in vitro and in vivo neurotrauma model systems based on laser transection and surgical nerve cut, we demonstrated that axotomy triggers actomyosin contraction coupled with calpain activity. As a consequence, neurons shrink acutely to force water out through aquaporin channels preventing swelling and bursting. Inhibiting shrinkage increased the probability of neuronal cell death by about 3-fold. These studies reveal a previously unrecognized cytoprotective response mechanism to neurotrauma and offer a fresh perspective on pathophysiological processes in the nervous system.Yüzüncü Yıl Universit

Sigara içenlerde dondurma-çözme işleminin zamana bağlı olarak sperm parametreleri, ince yapısı ve hücre ölümü üzerine etkileri

Sigara İçenlerde Dondurma-Çözme İşleminin Zamana Bağlı Olarak Sperm Parametreleri, İnce Yapısı ve Hücre Ölümü Üzerine EtkileriMehmet Şerif AYDINSigara içmek, sperm konsantrasyonunda düşüş, motilesinde azalma ve morfolojisinde bozulma meydana getirerek subfertiliteye neden olmaktadır. Sigara kullanımının sperm parametreleri üzerindeki etkileri bilinmesine rağmen, bunun kriyoprezervasyonla olan ilişkisi üzerinde yeterince durulmamıştır. Bu çalışma kriyoprezervasyonun sigara içen ve içmeyenlerin sperm parametreleri ve DNA fragmantasyonu üzerine olan etkilerini göstermeyi amaçlamaktadır. Semen örnekleri düzenli olarak sigara içen 20 kişi ve sigara içmeyen 21 kişi olmak üzere toplam 41 normospermik erkek gönüllüden alınmıştır. Semen örnekleri ikiye ayrılarak, ilk kısmı başlangıçtaki semen analizi için kullanılmıştır. İkinci kısma ise -196 oC’de sıvı nitrojen içerisinde 1 ay ve 3 ay süreyle dondurulup çözüldükten sonra aynı şekilde semen analizi yapılmıştır. Örnekler standart semen analizi ile morfoloji, motilite, konsantrasyon açısından ayrıca, TUNEL (TdT-mediated dUTP nick end labeling) yöntemi kullanılarak DNA fragmantasyon oranı açısından değerlendirilmiştir. Ultrastrüktürel analiz geçirimli elektron mikroskobu (TEM) ile yapılmıştır. Kriyoprezervasyon sonrası sperm morfolojisinin sigara grubunda daha çok etkilendiği görülmüştür. TEM incelemelerinde membran yapılarında bozulma ve artmış subakrozomal şişme görülmüştür. Başlangıçta DNA fragmantasyonu hücre oranı, sigara grubunda kontrol grubuyla aynı olmasına rağmen kriyoprezervasyon sonrasında her iki grupta da artışın yanı sıra sigara grubunda kontrol grubuna göre anlamlı bir artış saptanmıştır. Sonuç olarak kriyoprezervasyon membran yapılarında bozulma meydana getirmekte, sigara içenlerde sigara içmeyenlere kıyasla sperm morfolojisinde bozulmayı ve DNA fragmantasyonunu daha fazla arttırarak spermde olumsuz etkileri tetiklemektedir.Anahtar Kelimeler: Sperm, kriyoprezervasyon, sigara, DNA fragmantasyonu, ultrastrüktürTime Dependent Effects of Cryopreservation on Sperm Parameters, Ultrastructure and Cell Death in Cigarette SmokersMehmet Şerif AYDINSmoking causes subfertility due to sperm deterioration such as decreased concentration, impaired motility and abnormal morphology. Although evidence on the effect of smoking on sperm parameters is well known, its interference with cryopreservation is not clear. This study aims to investigate the effects of cryopreservation on sperm parameters and DNA fragmentation in smokers and nonsmokers. Semen samples were obtained from 41 normospermic male volunteers of whom 20 were smokers and 21 nonsmokers. The samples were separated into two aliquots. The first aliquot was kept for initial semen analysis. The second was stored in liquid nitrogen at -196oC, and analyzed following thawing after 1 and 3 months. Samples were analyzed in terms of motility, concentration and morphology with light microscope, and DNA fragmantation using TdT-mediated dUTP nick end labeling (TUNEL) by fluorescence microscope. Ultrastructural alterations were investigated with transmission electron microscope (TEM). Sperm morphology seemed to be affected more after cryopreservation in samples obtained from smokers. TEM investigations showed alteration in integrity of the membranes and increased subacrosomal swelling. Before freezing, the increase in DNA fragmentation rate in smokers was not statistically significant compared to that of nonsmokers. However, after thawing, the DNA fragmentation rates were significantly high in both smokers and nonsmokers compared to their respective rates before freezing. The extent of the increase in DNA fragmentation rate was significantly higher in smokers after thawing compared to that of nonsmokers. In conclusion, cryopreservation causes alterations in integrity of the membranes and increases DNA fragmentation, thus triggering relatively negative effects on the sperm samples of smokers compared to that of nonsmokers.Key Words: Sperm, cryopreservation, smoking, DNA fragmentation, ultrastructur

Far-infrared treatment of experimental sinusitis in rabbits: A pilot study

OBJECTIVE: Thermal therapy has existed for thousands of years and dates back to ancient civilizations such as the Finns, Romans, ancient Chinese, American Indians, and Egyptians. Sun therapy is a form of natural Far Infrared Rays (FIR) thermal therapy. Far infrared rays are invisible rays with the longest wavelength of natural sunlight. However, sunbathing should only be done in moderation, as sunlight contains ultraviolet rays that can burn and damage the skin. More than 30 million people in Asia, Europe, and Australia receive Far Infrared Therapy with FIR lamps. They emit 2-25-micron wavebands. We aimed to investigate whether FIR therapy that uses a similar waveband improves sinusitis symptoms. MATERIALS AND METHODS: This study was conducted on 10 New Zealand rabbits of both sexes, 7-8 months old, with an average weight of 1.8-2.4 kg. They were divided into four study groups. Sinusitis was induced in rabbits by blocking the sinus ostium, whereas some were left as control. Far infrared treatment with an infra-sin device was given with or without antibiotics. Mucosal samples were evaluated microscopically regarding epithelial eruption, epithelial vacuolization, interstitial edema, vascular congestion, leukocyte infiltration, and mucosal thickening. RESULTS: We found clinical and histopathological improvements in rabbits with sinusitis on the right side of the nose, to which we applied FIR treatment with the device Infra-Sin. CONCLUSIONS: Our findings in this animal study led us to conclude that it would be beneficial to study the effects of FIR treatment on people with allergic rhinitis and sinusitis and to identify the optimal dosing, treatment duration, and intensity. To add to our findings, looking into whether FIR therapy can be used as an alternative to antibiotic treatment for human sinusitis is essential

Effect of grape seed extract on bone formation in the expanded inter-premaxillary suture

Objective: Widening the inter-maxillary suture is a preferred procedure in orthodontics. However, relapse can occur in some patients after maxillary expansion therapy. This study aimed to increase bone formation in the inter-maxillary suture and decrease retention time with the help of grape seed extract (GS), which can stimulate bone tissue formation.Materials and Methods: A total of twenty rats were separated into the following two groups (n=20): the control group (group C) comprised rats that received only maxillary expansion therapy, and the other group (group GS) comprised rats that received maxillary expansion therapy along with GS. The grape seeds were systemically administered using an orogastric tube after maxillary expansion in group GS. Expansion springs were placed and activated to produce force. The springs were taken from the mouth 5 days later and exchanged with short rectangular wires for retention. This retention wire was placed for 12 days.Results: Significant differences were found in the percentage of newly formed bone (p<0.05) and bone area (p<0.05) between the two groups. Moreover, group GS had better bone formation than group C.Conclusion: Systemic use of GS during the stages of the orthopaedic expansion of the inter-premaxillary suture area increases newly formed bone and decreases the retention time

Transfer and integration of breast milk stem cells to the brain of suckling pups

WOS: 000445336600057PubMed ID: 30250150Beside its unique nutritional content breast milk also contains live cells from the mother. Fate of these cells in the offspring has not been adequately described. In this study, we aimed to detect and identify maternal cells in the suckling's blood and the brain. Green fluorescent protein expressing transgenic female mice (GFP+) were used as foster mothers to breastfeed wildtype newborn pups. One week and two months after the birth, blood samples and brains of the sucklings were analyzed to detect presence of GFP+ cells by fluorescence activated cell sorting, polymerase chain reaction and immunohistochemistry on the brain sections and optically cleared brains. The tests confirmed that maternal cells were detectable in the blood and the brain of the pups and that they differentiated into both neuronal and glial cell types in the brain. This phenomenon represents breastfeeding - induced microchimerism in the brain with functional implications remain to be understood.The Scientific and Technological Research Council of Turkey (TUBITAK) [114R078]This project was supported by "The Scientific and Technological Research Council of Turkey" (TUBITAK) with a grant number 114R078

Effects of a thermosensitive in situ gel containing mometasone furoate on a rat allergic rhinitis model

WOS: 000432900700002PubMed ID: 29644886Background: Mometasone furoate, one of the second generation intranasal corticosteroids, is currently used in suspension form due to its poor solubility. However, this is not favorable for nasal application because of the rapid elimination of the instilled drug from the nasal cavity by mucociliary clearance and delayed onset of action due to the slow dissolution of drug in suspension. Objective: The aim of this study was to determine the antiallergic effects of mucoadhesive thermosensitive in situ gel containing mometasone furoate that we developed previously to prolong the contact between the drug and nasal mucosa and to prevent drainage of the formulation in an ovalbumin-induced rat model of allergic rhinitis. Methods: An experimental allergic rhinitis model was developed in female Wistar albino rats by intraperitoneal injection of ovalbumin every 2 days for 14 days followed by its repeated intranasal instillation for 7 consecutive days. Intranasal instillation of ovalbumin was continued every other day for 14 days. Mometasone furoate in situ gel (5 mu g/10 mu l), mometasone furoate suspension (5 mu g/10 mu l), and physiological saline (10 ml) were administered into the bilateral nasal cavities from day 22 to day 35. Antiallergic effects were evaluated through histopathological evaluation, analysis of ovalbumin-specific serum immunoglobulin E, and a symptom score. Results: Mometasone furoate in situ gel significantly decreased the nasal symptoms and ovalbumin-specific serum immunoglobulin E level as compared with mometasone furoate suspension and physiological saline. Additionally, inflammatory histological symptoms such as mucosal edema, vascular dilatation, eosinophil infiltration, and loss of cilia within the nasal mucosa of allergic rhinitis model rats were remarkably improved with the treatment of mometasone furoate in situ gel. Conclusion: These results suggest that mometasone furoate in situ gel has a better therapeutic potential for the treatment of allergic rhinitis compared to mometasone furoate suspension.Scientific Research Project Coordination Unit of Istanbul University [41584]; TUBITAK (The Scientific & Technological Research Council of Turkey) [114S820, 1002]The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by Scientific Research Project Coordination Unit of Istanbul University (Project Number: 41584) and Rapid Support Program (1002) of TUBITAK (The Scientific & Technological Research Council of Turkey (Project Number: 114S820)

Therapeutic effects of metformin for noise induced hearing loss

Objective: This study aimed to investigate the healing effect of metformin on noise induced hearing loss (NIHL) by measuring audiological, biochemical and histological parameters.Materials and methods: 32 rats were divided into four groups (Group 1: Noise, Group 2: Noise + Metformin, Grup 3: Metformin, Grup 4: Control). Broadband noise was applied to Group 1 and Group 2 after basal measurements. Measuring audiological (distortion product otoacoustic emission (DPOAE) and Auditory Brainstem Response (ABR)), biochemical (total antioxidant status (TAS), total oxidant status (TOS), oxidative status index (OSI), DNA damage, IL-1 beta, IL-6, TNF alfa, HSF-1 and COX-2) and histological parameters.Results: Group 2 had significant decreases in ABR thresholds on day 7 and day 14 compared to day 1. DPOAE values of Group 2 on the 7th and 14th days were significantly higher than the post-noise levels. DNA damage, TOS and OSI values of Group 1 were significantly higher than the other groups. The Cox-2 value of Group 1 was higher than all other groups. The HSF-1 value of Group 2 was significantly higher than that of Group 1. In terms of IL-1 Beta, IL-6 and TNF-alpha values, there was no significant difference between groups 2, 3 and 4 and these values were significantly lower than group 1. In histopathological results of our study, no significant difference was found between the groups being exposed to noise and the control group.Conclusion: This study showed that early period of Metformin treatment has therapeutic effect on NIHL

The effects of human amniotic membrane on silicone related capsule formation in rats

Silicone breast implants are commonly used materials in plastic surgery for breast augmentation and reconstruction and the most severe complication of silicone implants are capsule contraction which occurs in 40% of patients. The aim of our study is to evaluate how the amniotic membrane alters the capsule formation effects of silicone 24 wistar rats were used in the study. We placed a bare silicone block into the left side (Subgroup A) and single layer amniotic membrane coated silicone block into the right side (Subgroup B) of the rats back. The rats were then separated into three groups and in group 1 rats were euthanized after 3 weeks, in group 2 after 12 weeks and in group 3 after 24 weeks. Then capsule thickness, fibroblast and lymphocyte cell counts were evaluated for each sample. In Group 2 and group 3, the capsule thickness in Subgroup B was detected to be statistically significantly lower than that in Subgroup A. In Group 1, 2, and 3, the lymphocyte count in the capsule tissue taken from Subgroup B was lower than Subgroup A but the difference was not statistically significant. In Group 2 and 3, the fibrocyte count detected in the capsule tissue in Subgroup B was found to be statistically significantly lower than Subgroup A. the amniotic membrane was demonstrated to reduce capsule thickness by the antifibrinolytic effect in our study