The influence of functional pinealectomy and exogenous melatonin application on healing of a burr hole in adult rat calvaria: a histological and immunohistochemical study.

BACKGROUND: Even today, repair of the cranial defects still represents a significant challenge in neurosurgery and various options have been used for their reconstruction to date. However, there are very few studies investigating the effects of exogenous administration of melatonin (MEL) as an agent that promotes bone regeneration. The goal of this study was to investigate the effects of functional pinealectomy (Px) and exogenous MEL administration on the bone repair properties and surrounding connective tissue alterations in a rat calvaria model. MATERIALS AND METHODS: The total of 30 adult female Wistar-Albino rats was randomly divided into three groups (n = 10): control group (CO; 12 h light/12 h dark exposure), functional Px group (24 h light exposure, light-induced functional Px), and Px+MEL group (light-induced Px + MEL, 20 mg/kg/day for 12 weeks). Critical-sized burr-hole defects (diameter: 3.0 mm) were surgically created by a single operator in the calvarium of all rats, using an electric drill. Animals in Px+MEL group received MEL 20 mg/kg/day for 12 weeks. At the end of the study, bone healing and connective tissue alterations surrounding drilled defect area in the rat calvaria were determined in haematoxylin-eosin-stained and Mallory Azan slices applied in anti-bone sialoprotein. Image Pro Express 4.5 programme was used for histomorphometric calculation of areas of new bone and fibrotic tissue. Normality control was performed by Shapiro-Wilk test. Variance homogeneities were examined by Shapiro-Wilk and Levene tests; Tukey HSD test was used as a post hoc method since there was no homogeneity problem. All hypothesis tests were performed at the 0.05 significance level. RESULTS: Histological analysis showed that the bone repair process in the Px+MEL group was similar to that of the CO group, whereas the functional Px group showed a delay. Histomorphometrically, it was found that the Px group had the largest hole diameter and the most fibrotic scar area, although no binary statistical significance was found between the CO and Px+MEL groups (p = 0.910). In terms of vascularisation, it was observed that the most vascular structure was found in the Px+MEL group among the scar tissue and ossification areas, while the vascularisation was the least in the Px group (p < 0.001). CONCLUSIONS: Our findings revealed that bone repair process was impaired in functional Px group, but exogenous MEL replacement was able to restore this response. Thus, it is concluded that utilisation of MEL may improve the bone repair in calvarial defects.

Unbiased estimation of the calcaneus volume using the Cavalieri principle on computed tomography images.

The size and shape of tarsal bones are especially relevant when considering some orthopedic diseases such as clubfoot. For this reason, the measurements of the tarsal bones have been the subject of many studies, none of which has used stereological methods to estimate the volume. In the present stereological study, we estimated the volume of calcaneal bone of normal feet and dry bones. We used a combination of the Cavalieri principle and computer tomographic scans taken from eight males and nine dry calcanei to estimate the volumes of calcaneal bones. The mean volume of dry calcaneal bones was estimated, producing mean results using the point-counting method and Archimedes principle being 49.11+/-10.7 or 48.22+/-11.92 cm(3), respectively. A positive correlation was found between anthropometric measurements and the volume of calcaneal bones. The findings of the present study using the stereological methods could provide data for the evaluation of normal and pathological volumes of calcaneal bones.

Immunohistochemical detection of insulin-like growth factor-I, transforming growth factor-beta2, basic fibroblast growth factor and epidermal growth factor-receptor expression in developing rat ovary.

The aim of this study was to determine the immunohistochemical expression and localization of insulin-like growth factor-I (IGF-I), transforming growth factor-beta2 (TGF-beta2), basic fibroblast growth factor (bFGF) and epidermal growth factor-receptor (EGF-R) in developing rat ovaries. Eighteen female Wistar rats were enrolled in this study; newborn (n=6), one-month-old (n=6) and adult (n=6) rats. Formalin-fixed and parafin-embedded ovarian tissues were stained with antibodies against IGF-I, TGF-beta2, bFGF and EGF-R, immunohistochemically. The ovarian cells were evaluated by semi-quantitative scoring system under light microscope. The staining of IGF-I, TGF-beta2, bFGF and EGF-R were most intense in the oocytes and were heavily at one-month-old rats. A moderate immunostaining in theca cells and corpus luteii reacted with IGF-I in adult rats. Furthermore the staining intensity for IGF-I was moderate in granulosa cells of newborn rat ovaries. We detected also a moderate staining for TGF-beta2 in corpus luteii of adult rats. In addition, we found a bFGF immunostaining mainly in oocytes of follicles of young and adult rats. Immunostaining for EGF-R was moderate in granulosa cells of one-month-old rats. In conclusion, this study suggests that growth factors play a pivotal role in ovarian function, especially in follicular development. The role of growth factor in controlling degeneration or growth (or both) of ovary follicles remain as explained.

Expression and localisation of Bcl-2 and Bax proteins in developing rat ovary.

Bcl-2 and Bax proteins localised mainly in granulosa cells. Primordial and primary follicles of new born rat ovary showed an intensive nuclear staining for Bax but faint staining for Bcl-2. In terms of staining intensity, no remarkable difference was observed within the same stage of developing follicle. Compared to new born rats, granulosa cells of adult and one month old rat ovary showed an increased staining both for Bcl-2 and Bax proteins. No staining was observed in primordial follicles of one month old and adult rats. However, granulosa cells of primary follicles, granulosa cells and theca cells in tertiary follicles of adult rat ovary also showed a strong staining for Bcl-2 and Bax proteins. Oocytes of follicles from different developmental stages revealed an apparent staining both for Bcl-2 and Bax proteins. However, in the more mature follicles oocytes stained more intensively. In developing corpus luteum a remarkable staining was observed for Bax. However, the staining was more prominent in regressing corpus luteum. Contrary to this, Bcl-2 stained the luteal cells in developing corpus luteum strongly, while in the fully developed corpus luteum no staining for Bcl-2 was observed. In conclusion, there was an apparent relation between the expression of the apoptosis regulating protein Bcl-2 and Bax and follicular development. Thus, during the follicular development Bcl-2 and Bax may be involved in granulosa cell demise in rat ovary. Furthermore, increased levels of Bax and decreased levels of Bcl-2 in the fully developed corpus luteum suggest that Bax plays a role in apoptosis of luteal cells in rat ovary.

Comparison of four methods for the estimation of intracranial volume: a gold standard study.

Investigators can infer how much reduction in volume has occurred since brain volume was at its peak, by combining measures of brain volume with measures of intracranial volume (ICV). Several methodologies have been proposed to asses the ICV. However, we have not seen a gold-standard study evaluating the results of the methodologies for the assessment of ICV. In the present study, the actual intracranial volume of 20 dry skulls was measured using the water-filling method, using this as a gold standard. Anthropometry, cephalometry, point-counting, and planimetry techniques were applied to the same skulls to estimate the ICV. Anthropometric and cephalometric measurements were carried out directly on skulls and roentgenograms, respectively. Consecutive computed tomography sections at a thickness of 10 mm were used to estimate the ICV of the skulls by means of the point-counting and planimetry methods. The mean (+/-SD) of the actual ICV measured by the water-filling method was 1,262.0 +/- 160.4 cm(3) (1,389.5 +/- 96.5 cm(3) for males and 1,134.5 +/- 94.3 cm(3) for females, respectively). Our results showed that the estimated values obtained by all four methods differed from the actual volumes of the skulls (P < 0.05). The data obtained by anthropometry resulted in overestimation. However, cephalometry, point-counting, and planimetry methods produced underestimation. After calibration, there were no significant differences between the actual volumes and the results of the four methods (P > 0.05). While the anthropometric method is easy and quick to apply, its result may deviate from the actual values. The optimized stereological techniques of point-counting and planimetry methods may provide unbiased ICV results since they take the third dimension of the structures into account.

Effects of constant lightness, darkness and parachlorophenylalanine treatment on tail regeneration in the lizard Ophisops elegans macrodactylus: macroscopic, biochemical and histological changes.

A better understanding of regenerative growth is very important for the development of new potential strategies. Recently, the pineal secretory product melatonin was shown to stimulate the regeneration process. In this study, we carried out an experimental investigation of tail regeneration in young adult lizards, Ophisops elegans macrodactylus Berthold, 1842, addressing the role of melatonin on the regeneration process. Lizards were divided into three groups: constant light-exposed control group (n = 13), constant dark-exposed group (n = 15) and parachlorophenylalanine (p-CPA) treatment group exposed to constant light (n = 15). Using a tail autotomy procedure, the effects of p-CPA treatment on connective tissue together with neural tissue and vascular tissue in regenerating tail in lizards were investigated. p-CPA (400 microg/kg body mass) was injected from day 0 to 30 days after autotomy. p-CPA treatment produced a significant increase in the length of the regenerated tail compared with light-exposed control and dark-exposed lizards. Total collagen content was found to be higher in p-CPA-treated animals in comparison with other groups. Histologically, a higher percentage of connective tissue and vascular tissue and a lower percentage of neural tissue were found in the regenerated tails of the p-CPA-treated lizards. Importantly, the percentage of neural tissue in lizards in the dark-exposed animals was higher than that in animals of both light-exposed and p-CPA treatment groups. Thus, it is clear that p-CPA has a stimulatory influence on fibroblast collagen production and vascularization of the regenerated tail in the lizards. Furthermore, it seems that the neural regeneration process was markedly enhanced in lizards exposed to continuous darkness. Based on the results of our study, it is suggested that melatonin may be an active factor that speeds up the rate of tail regeneration in lacertilians.

Morphometry of the sacrum for clinical use.

Degenerative disease and instability in the lower lumbar spine may necessitate fusion and stabilization supplemented by instrumentation to the sacrum. However, screw placement in a reasonable position is more difficult to achieve because of the unique anatomy of the first sacral (S1) vertebra. Therefore, this study has been conducted to evaluate sacrum anatomy of the Western Anatolian population in terms of morphometric measurements and make a comparison with previous studies as well as giving a guidance to the surgeons. In this study, 60 dry adult sacrums (30 male and 30 female) were assessed for morphometric analysis. The measurement data for the sacrum and S1 vertebra revealed that there was no significant difference between both sexes except the sacral width and sacral canal width (p<0.05). In the present study, the ratio of S1 corpus' width to sacral width was lower in females compared with males. A detailed knowledge of the morphometric data about sacrum is very important for spinal surgery, as pedicle screw insertion is crucial in spinal instrumentation in order to prevent neurological injury and/or fixation failure.

Extremely rare type of conjoined twins: cephalothoracopagus deradelphus.

Occurrence of conjoined twins (CT) with a chromosomal abnormality is a very rare event. The case presented is that of a cephalothoracopagus Deradelphus twin with one cerebrum and two brain stems joined at the cranial end of the midbrain. This rare condition and its diagnosis and management are discussed with regard to counselling and obstetrical care.