No abstract

RANKL and OPG are expressed in apical periodontitis, suggesting a role for these molecules during lesion development. However, the profiles of RANKL/OPG expression in periapical lesions remain unknown. In this study we investigated the patterns of RANKL and OPG mRNA expression by RealTime-PCR in human periapical granulomas (N=44), and compared them with sites presenting characteristic bone resorpting activity: healthy (n=14) and orthodontically stretched and compressed periodontal ligament (n=26), healthy gingiva (n=24), chronic gingivitis (n=32) and chronic periodontitis (n=34) samples. Both RANKL and OPG mRNA expression was higher in periapical granulomas when compared to healthy periodontal ligament. Distinct patterns of RANKL and OPG expression ratio were found in the granulomas and in different physiological and pathological conditions with characteristic bone resorption activity potentially being an indicative of the stable or progressive nature of the lesions. Lesions with radiographic image smaller than 5mm demonstrated higher RANKL/OPG expression than images greater than 5mm. Periapical granulomas presented heterogeneous patterns of RANKL and OPG expression, ranging from samples with RANKL/OPG ratio similar to that seen in sites with minimal or absent bone resorption, until samples with RANKL/ OPG expression pattern comparable with active bone resorption sites.

In order to improve the efficacy of conventional radiotherapy, attention has been paid to immune cells, which not only modulate cancer cell response to therapy but are also highly recruited to tumours after irradiation. Particularly, the effect of ionizing radiation on macrophages, using therapeutically relevant doses, is not well understood. To evaluate how radiotherapy affects macrophage behaviour and macrophage-mediated cancer cell activity, human monocyte derived-macrophages were subjected, for a week, to cumulative ionizing radiation doses, as used during cancer treatment (2 Gy/fraction/day). Irradiated macrophages remained viable and metabolically active, despite DNA damage. NF-kappaB transcription activation and increased Bcl-xL expression evidenced the promotion of pro-survival activity. A significant increase of pro-inflammatory macrophage markers CD80, CD86 and HLA-DR, but not CCR7, TNF and IL1B was observed after 10 Gy cumulative doses, while anti-inflammatory markers CD163, MRC1, VCAN and IL-10 expression decreased, suggesting the modulation towards a more pro-inflammatory phenotype. Moreover, ionizing radiation induced macrophage morphological alterations and increased their phagocytic rate, without affecting matrix metalloproteases (MMP)2 and MMP9 activity. Importantly, irradiated macrophages promoted cancer cell-invasion and cancer cell-induced angiogenesis. Our work highlights macrophage ability to sustain cancer cell activities as a major concern that needs to be addressed to improve radiotherapy efficacy.

The meiotic spindle and the zona pellucida exhibit molecular order when imaged with polarized optics. This study aimed to investigate possible factors contributing to the zona pellucida birefringence score and meiotic spindle visualization, and to evaluate whether these structures may predict intracytoplasmic sperm injection outcomes. Oocytes were divided into groups according to zona pellucida birefringence and meiotic spindle visualization. In addition, the cycles were split into three groups based on the zona birefringence of transferred embryos. A positive correlation was observed between zona birefringence and meiotic spindle visualization. In addition, when the meiotic spindle was observed, the fertilization rate among oocytes with high or low zona pellucida birefringence was similar. Implantation and pregnancy rates were significantly higher when embryos derived from high birefringence oocytes were exclusively transferred (P = 0.041 and P = 0.004 respectively). Furthermore, the miscarriage rate was higher when embryos derived from low birefringence oocytes were exclusively transferred. On the other hand, the total dose of FSH negatively affected meiotic spindle visualization. Results show that selection of embryos based on zona pellucida and meiotic spindle imaging can significantly improve implantation and pregnancy rates. Moreover, the dose of FSH used for ovarian stimulation may affect the organization of the oocyte meiotic spindle.

The development of a modified intracytoplasmic sperm injection (ICSI), called intracytoplasmic morphologically selected sperm injection (IMSI), demonstrated that a profound morphological investigation of the spermatozoon, under the magnification of 6600 x, enables outcome improvement. The aim of this study was to compare ICSI outcome with IMSI outcome. The meta-analysis results demonstrated no significant difference in fertilization rate between ICSI and IMSI groups. However, a significantly improved implantation (odds ratio (OR) 2.72; 95% confidence interval (CI) 1.50-4.95) and pregnancy rate (OR 3.12; 95% CI 1.55-6.26) was observed in IMSI cycles. Moreover, the results showed a significantly decreased miscarriage rate (OR 0.42; 95% CI 0.23-0.78) in IMSI cycles as compared with ICSI cycles. This is the first meta-analysis of published data to evaluate the potential benefits of IMSI. The pooled data of IMSI cycles demonstrate a statistically significant improvement in implantation and pregnancy rates and a statistically significant reduction in miscarriage rates. However, more randomized controlled trials are needed to confirm these results.