Histological and morphological development of the prepuce from birth to prepubertal age.

PURPOSE: To study the histological changes of the preputial tissue from birth to prepubertal age in order to define unnoticed morphological changes. MATERIALS AND METHODS: Prepuce samples were obtained from 79 healthy boys who underwent routine ritual circumcision. Specimens were divided into six groups according to the boys' age: newborn, 0-1 year of age, 2-3 years of age, 4-5 years of age, 6-7 years of age, and 8-9 years of age. Histologic analysis of the specimens was performed by H&E, Masson's trichrome, Verhoeff-Von Gieson, immunohistochemical staining. RESULTS: Microscopic examinations showed that average epithelial thickness increased after the neonatal period (p=0.001). When collagen fiber density was evaluated, no significant differences between groups were found (p=0.083). When the elastic fibers in the dermis were evaluated, it was determined that the number and thickness of elastic fibers increased with age. Immunohistochemical examinations showed that the number of peripheral nerves marked with S100 was lower in the neonatal period than at other ages (p=0.048). When the vessels marked with CD105 antibody were counted, there was no significant difference between the groups (p=0.078). CONCLUSIONS: This is the first study to examine the age-related structure of connective tissue elements in the foreskin. Our results showed that the prepuce's prepubertal maturation process is continuous, and the first 2 years of life are appropriate not only in relation to the physiological effects of age but also the optimum structural changes for wound healing, such as vessel diameter, epithelium thickness, peripheral nerve count.

CREB1 and PPAR-α/γ Pathways in Hepatic Ischemia/Reperfusion: Route for Curcumin to Hepatoprotection.

Background: Hepatic ischemia/reperfusion injury is a major problem that can exacerbate complications, particularly in liver transplantations. Objectives: This study aimed to investigate the cellular mechanisms of ischemia/reperfusion injury and hepatoprotection by curcumin. Methods: Wistar albino rats were divided into four groups as Control, Sham, I/R, and Cur+I/R. Hepatic ischemia/reperfusion was induced in I/R and Cur+I/R animals, the latter of which was also given 50 mg/kg/day of curcumin for 14 days. Liver aminotransferases and the transcription regulators of inflammation (RelA, IκB, PPAR-α, PPAR-γ, CREB1) were examined along with the histological examination. Results: Hepatic ischemia/reperfusion was found to disrupt hepatic microstructure and downregulate PPAR-α, PPAR-γ, and CREB1 transcripts. Curcumin supplementation in hepatic ischemia/reperfusion recovered the structural organization and promoted the hepatocyte regeneration while increasing expressions of PPARs and CREB1. RelA and IκB were found unaltered, possibly due to the crosstalk between targeted transcripts by ischemia/reperfusion and curcumin. Conclusions: In sum, PPAR-α/γ and CREB1 were involved in hepatic ischemia/reperfusion and, moreover, were detected to be stimulated by curcumin. PPAR and CREB pathways were found to provide a route to hepatoprotection for curcumin supplementation as evidenced by the microstructural improvement.

Perifosine and vitamin D combination induces apoptotic and non-apoptotic cell death in endometrial cancer cells.

Endometrial cancer is the most common cancer of the female reproductive system. Combination treatment with specific agents has been widely used as a targeted therapy for cancer. In this study, we aimed to investigate the anti-proliferative and apoptotic effects of varying concentrations of perifosine and vitamin D on the human endometrial cancer cell line (HEC-1A). HEC-1A cells were exposed to perifosine (10 µM, 30 µM), vitamin D (50 nM, 200 nM) and combinations of both for 48 h and 72 h. Monitoring of cell proliferation in a time-dependent manner was performed with the xCELLigence RTCA DP system. The levels of BCL2, BAX and P53 mRNA expression were examined using RT-qPCR. Apoptosis was determined using Annexin V, which were followed by flow cytometry analysis. Ultra-structural morphology of cells was analyzed by transmission electron microscopy (TEM) for 72 h. The anti-proliferative and apoptotic effects of the perifosine+vitamin D combination (30 µM + 200 nM at 48 h and 10 µM + 200 nM at 72 h) on HEC-1A cells were higher than in perifosine and vitamin D alone. It was observed that perifosine has increased the expression of BAX mRNA in HEC-1A cells in a dose-dependent manner. While perifosine+vitamin D combinations increased P53 mRNA expression in HEC-1A cells we did not find any significant change in BCL2, BAX mRNA expression levels. In TEM examinations of HEC-1A cells, perifosine appeared to lead autophagic cell death, whereas vitamin D caused paraptosis-like cell death and combination of perifosine+vitamin D caused apoptotic and non-apoptotic (paraptotic, autophagic and necrotic) cell death. Therefore, it is considered that the combination of both drugs in the treatment of endometrial cancer might be an alternative and effective treatment option through activating the apoptotic and non-apoptotic cell death mechanisms in cancer cells.

Effects of different cadaver preservation methods on muscles and tendons: a morphometric, biomechanical and histological study.

No prior publication studying the biomechanical and histological properties of cadavers fixed with Logan or modified Logan solution (MLS) was found in the literature. It was aimed in this study to compare MLS fixation and other cadaver preservation procedures regarding the use in basic histological studies, anatomy education and surgical trainings. This study was placed on 35 male 17-week-old Wistar Albino rats. MLS fixated tissues were systematically compared with 10% formalin (F10), saturated salt solution (SSS), Thiel and frozen/thawed (FT) tissues. Organoleptic (color, appearance, flexibility, odor, etc.), morphometric (e.g. length, width and cross-sectional area), biomechanical (Young's modulus, stiffness, maximum load, etc.) and histological (tendon and muscle fiber integrity, nuclear prominence, blur in microscopy, etc.) analyses were conducted. Organoleptic properties of Thiel and SSS fixated muscles and tendons were better preserved than F10 and MLS. No significant difference was observed in gross morphometric properties (e.g. length, width and cross sectional area) following any of the cadaver and tissue preservation techniques. MLS and F10 was observed to increase the stiffness, Young's modulus and maximum load parameters of the tendons. Thiel and SSS fixated tendons had similar mechanical properties to fresh and FT tendons. No effect of fixation solutions on tendons is observed in the histological analysis regarding fiber integrity, nuclear prominence, blur in microscopy, shrinkage of tissues. Thiel solution was observed to distort fiber integrity, nuclear prominence and blur the microscopy of muscle tissue. Thiel and SSS fixated muscles and tendons were observed to absorb more stain with Masson's trichrome staining and appear as darker red. No muscle and tendon shrinkage due to fixative solutions was observed with our fixation method. Pondering the organoleptic (color, appearance, consistency, odor, etc.) and biomechanical analyses (stiffness, Young's modulus, etc.), Thiel and SSS fixed cadavers are more suitable for purposes as surgical trainings and development of new surgical procedures. However, the change in the micro-anatomical structure of the muscles, especially with the Masson's trichrome staining, caused by these two solutions should not be overlooked.

Evaluation of the stapedial tendon growth dynamic in human fetuses.

PURPOSE: The main objective of the study was to investigate the morphometric properties of the stapedial tendon (ST) for pediatric otosurgeons and anatomists. METHODS: The present study was placed on 15 fetuses (8 females, 7 males) aged from 20 to 30 weeks of gestation (at mean, 24.27 ± 3.24 weeks) using the collection of the Anatomy Department of Medicine Faculty, Mersin University. All measurements were obtained with a digital image analysis software. RESULTS: In terms of male/female or right/left comparisons, no statistically significant difference was found in relation with the numerical data of ST. The surface area, length, and width of ST were detected as follows: 0.61 ± 0.15 mm2, 1.27 ± 0.30 mm, and 0.45 ± 0.08 mm, respectively. The absence of ST was observed in two fetuses with and without severe malformations. In another fetus with cleft lip and polydactyly, multiple abnormalities were bilaterally identified in the middle ear: (1) the absence of the incudostapedial joint and (2) the presence of an abnormal tissue attaching to the stapes. The abnormal tissue was determined to be irregular dense connective tissue using light microscope and electron microscope. CONCLUSION: Our findings showed that ST did not proportionally grow according to increasing gestational weeks. In the light of the numerical data, we thought that similar to stapes, ST attains the adult size in the fetal period. As ST anomalies may accompany severe malformations (e.g., cleft lip, polydactyly or syndactyly) that can be easily detected on observation by clinicians, we suggest that the detailed examination of middle ear in newborns should be taken into account for early diagnosis of conductive hearing loss to prevent any management delays.

Anatomic and histological analyses of chiasma plantare and long flexor tendons of the foot on human fetuses.

PURPOSE: The aim of present study was to reveal slip transfers related to flexor hallucis longus (FHL) and flexor digitorum longus (FDL) by dissection and to investigate detailed structure of chiasma plantare composed of FHL-FDL tendons and quadratus plantae (QP), with precise composition of the long flexor tendons of lesser toes by histological sections in human fetuses. METHOD: Slip transfers related to FHL and FDL tendons were identified and the related morphometric measurements were taken with dissection in 28 formalin-fixed fetuses (25-40 weeks). Composition and restoration of chiasma plantare and long flexor tendons of lesser toes were traced histological by analyzing movements of the tissues on the sequential coronal sections in five fetuses in the third trimester. The numbers of layers constituting chiasma plantare and the muscles that formed layers were specified. Each of two to five flexor tendons arising from the chiasma plantare was analyzed regarding its formation and contribution of FHL slip. RESULTS: Slip transfers were found as FHL slip in 86% and cross-connections in 14%. The ratios of the slip width to that of FHL and FDL tendons were found higher than in adult literature. Variance in the involvement of slip to FDL and QP, formation and layering of chiasma plantare and formation of long flexor tendons from chiasma plantare were revealed and great similarities were found with data from dissection of adult in literature. CONCLUSION: Slip transfers between FHL and FDL tendons, and layering properties of chiasma plantare were largely finalized during intrauterine period, while structural changes in slip seem to continue in the later stages of life, possibly by the effects of growth and usage of the extremity. In addition to individual variations, investigating the contribution of FHL slip, FDL and QP to long flexor tendons by different methods in literature is also suggested to be responsible for some diversities of our histological study.

Effect of low-level 1800 MHz radiofrequency radiation on the rat sciatic nerve and the protective role of paricalcitol.

The nervous system is an important target of radiofrequency (RF) radiation exposure since it is the excitable component that is potentially able to interact with electromagnetic fields. The present study was designed to investigate the effects of 1,800 MHz RF radiation and the protective role of paricalcitol on the rat sciatic nerve. Rats were divided into four groups as control, paricalcitol, RF, and RF + paricalcitol. In RF groups, the rats were exposed to 1,800 MHz RF for 1 h per day for 4 weeks. Control and paricalcitol rats were kept under the same conditions without RF application. In paricalcitol groups, the rats were given 0.2 µg/kg/day paricalcitol, three times per week for 4 weeks. Amplitude and latency of nerve compound action potentials, catalase activities, malondialdehyde (MDA) levels, and ultrastructural changes of sciatic nerve were evaluated. In the RF group, a significant reduction in amplitude, prolongation in latency, an increase in the MDA level, and an increase in catalase activity and degeneration in the myelinated nerve fibers were observed. The electrophysiological and histological findings were consistent with neuropathy, and the neuropathic changes were partially ameliorated with paricalcitol administration. Bioelectromagnetics. 39:631-643, 2018. © 2018 Wiley Periodicals, Inc.

Effects of silica nanoparticles on isolated rat uterine smooth muscle.

In spite of their widespread use, toxicity of silica nanoparticles (SiO2 NPs) to mammalian has not been extensively investigated. In the present study, it is aimed to investigate the effects and the mechanism of action of 20 nm sized SiO2 NPs on isolated uterine smooth muscle. A total number of 84 preparations of uterine strips were used in the experiments. Study was designed as four groups: group I (control), group II (0.2 mM SiO2 NPs), group III (0.4 mM SiO2 NPs) and group IV (0.8 mM SiO2 NPs). Spontaneous contractions were recorded using mechanical activity recording system. Superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and malondialdehyde (MDA) levels were measured using the spectrophotometric methods. Apoptosis of the cells was detected using immunofluorescence staining assay. SiO2 NP distribution and ultrastructural changes were determined by transmission electron microscopy. In groups II-IV, the frequency of contraction was significantly lower than that of the group I, whereas the contraction energy significantly decreased only in group IV. SOD and GSH-Px activities were significantly lower in experimental groups compared to the control group. MDA level and apoptotic cells were significantly higher in all SiO2 groups compared to the control group. Numerous SiO2 NPs in cytoplasm and connective tissue were observed in all dose groups. These findings showed that 20 nm sized SiO2 NPs enter the connective tissue and cytoplasm of uterine muscle cells and cause oxidative stress and apoptosis leading to impaired uterine contractile activity.

GDF9 and BMP15 Expressions and Fine Structure Changes During Folliculogenesis in Polycystic Ovary Syndrome.

BACKGROUND: Polycystic ovary syndrome is the most frequently seen endocrine disorder in women of reproductive age with a prevalence of about 10%. AIMS: To investigate the efficiency of growth differentiation factor 9 and bone morphogenetic protein 15 during folliculogenesis in a dehydroepiandrosterone-induced mouse Polycystic ovary syndrome model. STUDY DESIGN: Animal experimentation. METHODS: Mice were divided into 3 groups: control, vehicle and Polycystic ovary syndrome. Polycystic ovary syndrome model mice were developed by the injection of dehydroepiandrosterone dissolved in 0.1 mL of sesame oil. Ovarian tissues were examined for growth differentiation factor 9 and bone morphogenetic protein 15 using immunofluorescent labelling and electron microscopic examinations. RESULTS: The immunoreactivity of growth differentiation factor 9 and bone morphogenetic protein 15 proteins decreased (p<0.05) in the Polycystic ovary syndrome group (27.73±8.43 and 24.85±7.03, respectively) compared with the control group (33.72±11.22 and 31.12±11.05, respectively) and vehicle group (33.95±10.75 and 29.99±10.72, respectively). Apoptotic changes were observed in granulosa cells, lipid vacuoles increased in Theca cells and thickening and irregularities were noted in the basal lamina of granulosa cells. An increased electron density in the zona pellucida in some of the multilaminar primary and secondary follicles in the Polycystic ovary syndrome model was also observed at the ultrastructural level. CONCLUSION: These results suggest that the decrease in the growth differentiation factor 9 and bone morphogenetic protein 15 expression initiated at the primary follicle stage effect the follicle development and zona pellucida structure and may cause subfertility or infertility in Polycystic ovary syndrome.