Several cellular mechanisms affect nuclear morphology which can therefore be used to assess certain processes. Here, we present an analytic tool to quantify the number of cells in a population that present characteristics of senescence, apoptosis or nuclear irregularities through nuclear morphometric analysis. The tool presented here is based on nuclear image analysis and evaluation of size and regularity of adhered cells in culture. From 46 measurements of nuclear morphometry, principal component analysis filtered four measurements that best separated regular from irregular nuclei. These measurements, namely aspect, area box, radius ratio and roundness were combined into a single nuclear irregularity index (NII). Normal nuclei are used to set the parameters for a given cell type, and different nuclear phenotypes are separated in an area versus NII plot. The tool was validated with β-gal staining for senescence and annexin or caspases inhibitor for apoptosis as well as several treatments that induce different cellular phenotypes. This method provides a direct and objective way of screening normal, senescent, apoptotic and nuclear irregularities which may occur during failed mitosis or mitotic catastrophe, which may be very useful in basic and clinical research.
Weight loss in morbidly obese patients was associated with a reduction in the prevalence of hepatic fibrosis.
The purpose of this study was to develop and validate a multimedia software application for mobile platforms to assist in the teaching and learning process of design and construction of a skin flap. Traditional training in surgery is based on learning by doing. Initially, the use of cadavers and animal models appeared to be a valid alternative for training. However, many conflicts with these training models prompted progression to synthetic and virtual reality models. Fifty volunteer fifth- and sixth-year medical students completed a pretest and were randomly allocated into two groups of 25 students each. The control group was exposed for 5 minutes to a standard text-based print article, while the test group used multimedia software describing how to fashion a rhomboid flap. Each group then performed a cutaneous flap on a training bench model while being evaluated by three blinded BSPS (Brazilian Society of Plastic Surgery) board-certified surgeons using the OSATS (Objective Structured Assessment of Technical Skill) protocol and answered a post-test. The text-based group was then tested again using the software. The computer-assisted learning (CAL) group had superior performance as confirmed by checklist scores (p<0.002), overall global assessment (p = 0.017) and post-test results (p<0.001). All participants ranked the multimedia method as the best study tool. CAL learners exhibited better subjective and objective performance when fashioning rhomboid flaps as compared to those taught with standard print material. These findings indicate that students preferred to learn using the multimedia method.
Oxaliplatin (OXA) is a platinum compound widely used in the treatment of some solid tumors, especially colorectal cancer. Despite its usefulness, oxaliplatin-associated neurotoxicity represents the main dose-limiting factor of this drug, and until now, there is no suitable treatment. Chemotherapy with oxaliplatin also increases the rate of developing hepatic damages with inflammatory activity, termed chemotherapy-associated steatohepatitis (CASH). In the present study, we aimed to compare the effects of a series of antioxidant compounds on simultaneous development of oxaliplatin-induced hepato- and neurotoxicity in mice. Mice BALB/c were treated with oxaliplatin for 6 weeks, 10 mg/kg, intraperitoneally, resulting in mechanical allodynia and hepatic steatosis. We administered the following antioxidant compounds--rutin (RT) (20 mg/kg), resveratrol (RVS) (100 mg/kg), quercetin (QT) (20 mg/kg), and quercetin nanoemulsion (NQT) (20 mg/kg)--daily by gavage to BALB/c, and N-acetylcysteine (NAC) was used as positive control. Treatments with RSV, RUT, or NQT were able to prevent mechanical allodynia when compared to the OXA group, and this effect was associated with decreased c-Fos immunopositivity in the lumbar spinal cord. Regarding the effects on steatohepatitis, RVS, QT, and NQT almost completely reversed the mean liver weight increase induced by OXA. In accordance with these previous data, histological evaluation indicated attenuation of all features of hepatic steatosis evaluated in RSV, RUT, QT, and NQT groups. These compounds were able to reduce the immunopositivity for the apoptosis marker caspase-3. On the other hand, only QT and NQT treatments were able to reduce neutrophil migration measured by myeloperoxidase (MPO) activity. These results suggest that the compounds tested, RSV, RUT, QT, and NQT, would be useful for the clinical treatment of neuro- and hepatoxicity induced by oxaliplatin.
This study aimed to evaluate the effect of final irrigation protocols on microhardness reduction and erosion of root canal dentin. Sixty root canals from mandibular incisors were instrumented and randomly divided into six groups (n = 10) according to the irrigant used: QMiX, 17% EDTA, 10% citric acid (CA), 1% peracetic acid (PA), 2.5% NaOCl (solution control), and distilled water (negative control). The chelating solutions were used to irrigate the canal followed by 2.5% NaOCl as a final flush. After the irrigation protocols, all specimens were rinsed with 10 mL of distilled water to remove any residue of the chemical solutions. Before and after the final irrigation protocols, dentin microhardness was measured with a Knoop indenter. Three indentations were made at 100 µm and 500 µm from the root canal lumen. Afterwards, the specimens were prepared for scanning electron microscopic analysis and the amount of dentin erosion was examined. Wilcoxon and Kruskal-Wallis tests were used to analyze the results with a significance level set at 5%. At 100 µm, all protocols significantly reduced dentin microhardness (p < .05), while at 500 µm, this effect was detected only in the EDTA and QMiX groups (p < .05). CA was the irrigant that caused more extensive erosion in dentinal tubules, followed by PA and EDTA. QMiX opened dentinal tubules, but did not cause dentin erosion. Results suggest that QMiX and 17% EDTA reduced dentin microhardness at a greater depth. Additionally, QMiX did not cause dentin erosion.
Heat shock protein (Hsp)-peptide complexes purified from tumors can prime the immune system against tumor antigens, but how they contribute to the generation of immune responses against naturally occurring tumors is unknown. Murine tumors expressing high amounts of Hsp70 are preferentially rejected by the immune system, suggesting that low Hsp70 expression is advantageous for tumor growth in the host. To determine whether Hsp70 was differentially expressed in human tumors, inducible Hsp70 expression was quantitatively (by Western blot) and qualitatively (by immunohistology) analyzed in 53 biopsies of tumor and normal breast tissue. The mean expression of inducible Hsp70 was significantly higher in tumor compared with normal tissue (U ϭ 899.0; P ϭ 0.0033). However, a significant negative association of the amount of Hsp70 expressed by tumor tissue was found with metastasis (r 2 ϭ Ϫ0.309; P ϭ 0.05). After 3 years, follow-up analysis determined that 7 of the 53 patients relapsed, and 5 died. Hsp70 expression in tumor (but not normal) cells was significantly lower in relapse patients and patients with metastatic disease than in patients with no relapse or metastasis. Together, these observations support the hypothesis that Hsp70 plays a role in tumor expansion in vivo, and tumors that downregulate it may be able to evade immunosurveillance and grow.
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