The present study explored three-dimensional spatial arrangements of the cardiomyocytes and microvessels within the heart of rats and evaluated the arrangement for differences after diabetes using second-order stereology. Isector method was applied to obtain isotropic uniform random sections of the heart. The pair correlation g(r) and cross-correlation functions were estimated by counting dipole probes (with length of 3.57μm) superimposed on histological sections of the heart. The co-variograms indicated that the curve of g(r) for the cardiomyocyte showed a gap between cardiomyocytes at r=21-25μm in the control rats and a wider gap at r=18-50μm in diabetic hearts. Estimates of g(r) for the vessels also showed a wider gap (at r=25-39μm) in diabetic hearts compared to the control rats (r=25-32μm). These indicate a negative correlation (repulsion) between the cardiomyocytes and microvessels in the diabetic hearts. Evaluation of the cross-correlation function of the cardiomyocytes and microvessels showed that at 32-36μm, both structures had a negative correlation in the control group, but not in the diabetic rats.

CONCLUSION:

Dissociation of the cardiomyocytes at some places can be seen in diabetic heart. This can be seen also in microvessels. Neither cardiomyocytes nor microvessels are arranged normally after diabetes.

Quantitative data obtained by means of design-based stereology can add valuable information to studies performed on a diversity of organs, in particular when correlated to functional/physiological and biochemical data. Design-based stereology is based on a sound statistical background and can be used to generate accurate data which are in line with principles of good laboratory practice. In addition, by adjusting the study design an appropriate precision can be achieved to find relevant differences between groups. For the success of the stereological assessment detailed planning is necessary. In this review we focus on common pitfalls encountered during stereological assessment. An exemplary workflow is included, and based on authentic examples, we illustrate a number of sampling principles which can be implemented to obtain properly sampled tissue blocks for various purposes.

Central nervous system development is controlled by highly conserved homeoprotein transcription factors including HOX and TALE (Three Amino acid Loop Extension). TALE proteins are primarily known as HOX-cofactors and play key roles in cell proliferation, differentiation and organogenesis. MEIS1 is a TALE member with established expression in the developing central nervous system. MEIS1 is essential for embryonic development and Meis1 knockout mice dies at embryonic day (E) 14.5. However, Meis1/MEIS1 expression in the devolving forebrain, at this critical time-point has not been studied. Here, for the first time we characterize the region-specific expression of MEIS1 in E14.5 mouse forebrain, filling the gap of MEIS1 expression profile between E12.5 and E16.5. Previously, we reported MEIS1 transcriptional regulatory role in neuronal differentiation and established forebrain-derived neural stem cells (NSC) for gene therapy application of neuronal genes. Here, we show the dynamic expression of Meis1/MEIS1 during the differentiation of forebrain-derived NSC toward a glial lineage. Our results show that Meis1/MEIS1 expression is induced during NSC differentiation and is expressed in both differentiated neurons and astrocytes. Confirming these results, we detected MEIS1 expression in primary cultures of in vivo differentiated cortical neurons and astrocytes. We further demonstrate Meis1/MEIS1 expression relative to other TALE family members in the forebrain-derived NSC in the absence of Hox genes. Our data provide evidence that forebrain-derived NSC can be used as an accessible in vitro model to study the expression and function of TALE proteins, supporting their potential role in modulating NSC self-renewal and differentiation.

The present study clarifies developmental organization of the oxytalan fiber system in the periodontal space of both the enamel (labial) and cementum (lingual) sides of rat incisors. The number of oxytalan fibers per unit area (μm(2)) was counted in rat incisors at stages of embryonic day 20 (E20) to postnatal day 35 (P35). Oxytalan fibers in the periodontal space of the enamel side were apt to decrease in number during the postnatal period, whereas their number remained almost unchanged on the cementum side during the developmental period. When the incisor emerged through the gum at P11, thinner oxytalan fibers distributed in the apical growing periodontium of the cementum side seemed to be fused with one another to become thicker fibers as has been reported for rat molars (Inoue et al., 2012). Thus, the oxytalan fiber system in the periodontal space represented significant differences in its distributional density between the enamel and cementum sides after E23. At the stage of P35, oxytalan fibers presented significantly denser distribution in all territories of the periodontal ligament of the cementum side versus the enamel side. The present findings claim that the oxytalan fiber system might bind the tooth to the periodontal ligament and provide equilibrium of vascular system and control of blood flow in the periodontal ligament of the cementum side, while it might exclusively regulate the high level of physiologically adapted vasculature in the periodontal space of the enamel side.

Implantation of non-articular (heterotopic) chondrocyte-based implants might be an alternative approach to articular cartilage repair. This strategy could be helpful in cases in which there are no or too few articular chondrocytes available. Therefore, this study was undertaken to compare joint cartilage defect healing in the minipig model after implantation of heterotopic auricular and orthotopic articular chondrocytes. Poly-glycolic acid (PGA) associated three-dimensional (3D) constructs were prepared culturing autologous minipig-derived articular and auricular chondrocytes for 7 days in a dynamic culture system. Chondrocyte PGA constructs were implanted into 8mm diameter and ~1.1mm deep chondral defects within the medial and lateral condyles of the minipig knee joints. Empty defects served as controls for assessment of the intrinsic healing response. Defect healing was monitored 6 months post implantation using a macroscopic and microscopic score system and biomechanical analysis. Neo-cartilage formation could be observed in the PGA constructs seeded with articular and auricular chondrocytes in vivo. The defect healing did not significantly differ at the macroscopic and histological level in response to implantation of either autologous articular or auricular chondrocytes seeded constructs compared with the empty defects. Although the differences were not significant, the auricular chondrocytes-based implants led to a slightly inferior repair quality at the macroscopic level, but a histologically superior healing response when compared with the empty defect group. However, biomechanical analysis revealed a higher stiffness in repair tissues produced by auricular chondrocyte implantation compared with the other groups. Deduced from these results, articular chondrocytes represent the preferable cell source for implantation.

The metameric pattern of the axial skeleton is established during embryogenesis by somite formation from the unsegmented paraxial mesoderm (presomitic mesoderm). Here, we have investigated the morphology of the anterior presomitic mesoderm of chick embryos using scanning electron microscopy. We found periodically arranged transverse clefts in the anterior region of the presomitic mesoderm. These gaps can be regarded as physical boundaries between prospective somites in the determined zone of the presomitic mesoderm. This study provides additional evidence suggesting that prospective somite boundaries are not only marked by defined zones of gene expression, but are also accompanied by changes in cellular morphology that give rise to identifiable morphological segments.

Periodontal ligament (PDL) cells, a major component of the tooth supporting apparatus, share osteoblastic characteristics including their responsiveness to parathyroid hormone (PTH). Clinical studies have already pointed to the benefit of PTH in supporting regenerative processes in the craniofacial region. However, those reports did not analyze which cells mediated the PTH effect on the alveolar bone. The aim of the present study has been to further elucidate the mechanism of action of continuous PTH application on human PDL-cells mimicking a local bolus application and to analyze its intracellular signalling pathways to widen the theoretical basis for future development of reliable local PTH delivery protocols. Analyses of PDL of extracted human teeth as well as cultured human PDL-cells demonstrated strong expression of PTH-receptor-1 by immune fluorescencecytochemistry/histochemistry. To examine the effect of short time continuous PTH treatment on PDL-cell osteogenic differentiation, PDL-cells were stimulated for 48h. Analyses for mRNA and protein expression of the early osteogenic marker alkaline-phosphatase revealed an enhanced expression. Pathways analyses mediating the PTH effect resulted in a similar effect when PDL-cells were stimulated with either the signal specific fragments lacking the PKA-activating domain PTH(3-34), PTH(7-34), second-messenger-analogues PKC (PMA) or inhibitors for PKA (H8). Inhibition of the PKC-dependent pathway by stimulation with PTH(1-31), PKA second-messenger-analogue (forskolin) or PKA-inhibitor (RO-32-0432) abolished the PTH effect These data indicate abundant expression of PTH1R within the PDL and a stimulatory effect of short time continuous PTH on PDL cell differentiation towards an osteogenic phenotype and suggested local PTH application protocols as a possible treatment option in periodontal therapy.

INTRODUCTION:

The Société Française de Chirurgie Digestive and the American Society of Colon and Rectal Surgeons recommend a ligation at the origin of the primary feeding vessel for sigmoid cancer to ensure optimal lymphadenectomy. We evaluated the correlation between the level of ligation defined by the surgeon and the real level of ligation visualized on postoperative CT scan.

PATIENTS AND METHODS:

From December 2004 to August 2010, in a series of 146 patients undergoing colectomy for sigmoid cancer, 51 (19 women) CT measurements (visualization of the left colonic artery (LCA), length of the arterial stump) were performed by a radiologist blinded to operative data.

RESULTS:

This series comprised 63% of men with a mean age of 69 years. A correlation was demonstrated between the level of ligation assessed by the surgeon and the real level of ligation demonstrated on postoperative CT scan in 41% of cases. No risk factors for absence of correlation were identified (laparoscopy, gender, BMI, emergency, and ASA score). In the "no correlation" group, the site of ligation was overestimated in 70% of cases. No significant difference was observed between the "correlation" and "no correlation" groups for lymphadenectomy (21.6 and 18 lymph nodes, p=0.5593) or 5-year overall survival (71.4 and 93.1 months, p=0.57).

CONCLUSION:

In conclusion, the surgical and radiological correlations are low as the intraoperative estimation of the level of IMA ligation was correlated with CT findings in less than 50% of cases. No risk factors for non-correlation were identified, and there was no impact on lymphadenectomy. Overestimation of the level of ligation was the most frequent situation but did not appear to have any impact on tumor staging or on patient management in this group of patients.

Whole-body multidetector computed tomography (CT) was performed on the mummified corpse of two-year-old Rosalia Lombardo, an anthropogenic mummy displayed in the Capuchin Catacombs of Palermo, Sicily, Italy. Rosalia Lombardo reportedly died of bronchopneumonia in 1920 and was preserved by the embalmer and taxidermist Alfredo Salafia with a formaldehyde-based fluid. Rosalia Lombardo's body is still exhibited in the Capuchin Catacombs inside the original glass-topped coffin in which she was placed. Only her head is visible: the rest of her body is covered by a sheet. CT images of Rosalia's body within her coffin were of reduced quality because of distinct metal artifacts caused by the coffin itself. Nevertheless, a detailed radiological analysis was possible for most of the body. Analysis of the data from the CT examination revealed indicators for the historically-reported endovasal and intracavity treatment. Rosalia's entire body was preserved in a remarkable state. The exceptional preservation of her internal organs made it possible to consider a radiological diagnosis of pneumonia. For this study, CT was determined to be the ultimate method for investigation, since Rosalia's body had to be kept untouched in her sealed coffin for conservation purposes. The CT examination offered new insights into the current preservation status of the body, and the superior contrast of CT allowed detailed assessment of different tissues. Post-processing methods provided reconstructions on any desired plane, as well as three-dimensional reconstruction, for the best possible visualization and interpretation of the body.