Poly(glycidyl methacrylate) modified cellulose nanocrystals and their PBAT-based nanocomposites.

Melt processing of cellulose nanocrystals (CNCs) reinforced nanocomposites is still a serious challenge due to the hydrophilic nature of CNCs and their severe agglomeration tendency within the polymer melt. In this study, chemical modification of CNC through grafting poly(glycidyl methacrylate) (PGMA) with various degrees was implemented. Wettability of the modified CNCs (mCNCs) were controlled and their structure was characterized through Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), optical microscopy, X-ray diffraction (XRD), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). The nanocomposites of polybutylene adipate terephthalate (PBAT) with 3 wt% CNC and mCNC were prepared using an internal melt mixer. To differentiate the effects of CNC and PGMA molecules on the final properties of nanocomposites, PBAT/PGMA compounds were separately prepared. To confirm the chain characterization and molecular weight of the synthesized PGMAs, 1H NMR and gel permeation chromatography (GPC) analysis were conducted. Melt rheological analysis, dynamic mechanical analysis (DMA), DSC, and atomic force microscopy (AFM) were used to monitor the mCNC dispersion quality and the effect of PGMA modification in PBAT compounds. The results revealed that grafting CNC with longer PGMA considerably improved the CNCs' dispersion quality within PBAT. Such dispersion enhancement of long-chain mCNCs and interfacial interaction of PGMA and PBAT resulted in a noticeable increase in storage modulus and complex viscosity of the final nanocomposites.

Could immune cells be associated with nephropathy in Fabry disease patients?

BACKGROUND: In Fabry Disease (FD), although the primary factor initiating kidney damage is glycosphingolipid accumulation, secondary conditions such as increased inflammation and fibrosis may cause this damage to progress. These processes may be induced by immune cells. Therefore, we aimed to investigate the peripheral lymphocyte subgroup analysis of the patients with FD and compare these results with healthy individuals. In addition, we performed T, B, NK, and plasma cell analyses in kidney biopsy materials and compared these kidney biopsy results with the biopsy results of patients whose kidney functions were impaired after 4 years of regular ERT. MATERIALS AND METHODS: 18 FD and 16 healthy individuals were included in the study. T-B lymphocyte and NK-cell populations were determined. We performed kidney biopsies (KBx) on 13 patients with FD prior to ERT. Of these, 4 patients had rebiopsy after 4 years of regular ERT. Immunohistochemical staining was performed to define immune cell infiltration. RESULTS: There was no statistically significant difference in terms of total, helper and cytotoxic T-lymphocyte and CD3-CD16+CD56+ natural killer (NK)-cell count (p = 0.20; p = 0.12; p = 0.76; p = 0.75, respectively).According to KBx findings prior to ERT, all patients had interstitial fibrosis (IF), podocyte vacuoles (PV), and podocyte inclusion (PI), CD3, CD4, CD8, CD16, and CD56 positivity at different levels. None of the patients had CD19, CD20, and CD138 positivity at the first biopsies. When we compared the first and the second KBx results of the two progressors, we also demonstrated that CD3+4+T-cells infiltration remained the same, whereas CD8+T cells, CD16+ and 56+NK-cells infiltration were significantly decreased. In contrast, CD20+B cells and CD138+plasma cell infiltration were significantly increased despite 4 years of ERT (15 fold and sixfold, respectively). The CD20+B and CD138+ plasma cells and IF were positively correlated with proteinuria. CONCLUSIONS: The progression of FD nephropathy and proteinuria is increased despite a long-term ERT. Immune cells, primarily B and plasma cells, might cause these unwanted consequences.

Colonic fibronectin and E-cadherin in mild ulcerative colitis.

BACKGROUND: Although fibronectin has an important role in wound repair, nearly no human studies to date have investigated its condition in ulcerative colitis (UC) histologically. E-cadherin plays a critical role in the repair of normal epithelial tissues. This study aims to find out the condition of these two molecules in UC. MATERIAL AND METHODS: The records of 22 UC patients during the period of 2004‒2009 were retrospectively analyzed. We also included 24 patients with sporadic colorectal cancer (SCC) and 24 patients with normal colonoscopic biopsies who served as the control group. Colonoscopic biopsies were stained with E-cadherin and fibronectin. RESULTS: The E-cadherin loss was significantly more prominent in the SCC group, followed by the UC group and control group. The situation was reverse for fibronectin. We also observed that while the E-cadherin loss was still ongoing in all of the endoscopically inactive cases, the fibronectin staining resembled the staining pattern of normal individuals in ten out of thirteen UC patients. CONCLUSION: We suggest that the decrease in E-cadherin, even in the inactive period, might be the cause of why UC is not just a compensatory change in repair of inflammation. The results of staining with fibronectin in UC patients were between normal individuals and SCC patients. Further studies are necessary to confirm our results (Tab. 2, Fig. 6, Ref. 15). Text in PDF www.elis.sk Keywords: ulcerative colitis, fibronectin, E-cadherin.

Diagnostic value of MDCT in determining the perinephric fat tissue and renal sinus invasion in patients with clear cell renal cell carcinoma.

AIMS: To investigate the accuracy of multidetector computed tomography (MDCT) findings, and the effect of tumor volume in determining the perinephric and renal sinus invasion in clear cell renal cell carcinomas (ccRCCs). METHOD: Fifty patients with ccRCCs underwent non-contrast and nephrographic-phase contrast-enhanced MDCT examination before total nephrectomy. The following MDCT features were used to diagnose perinephric fat tissue invasion: perinephric stranding, perinephric vascularity, and irregular contour. The following MDCT features were used to diagnose renal sinus fat invasion: elongation of tumor into renal sinus, invasion, or compression of pelvicalyceal system. Histopathologic examinations were used as a gold standard. RESULTS: Fourteen out of 50 ccRCCs patients (28%) had histopathological-proven perinephric fat tissue invasion. The sensitivity, specificity, PPV, NPV, and accuracy of MDCT in the detection of perinephric fat tissue invasion were found 64%, 58%, 38%, 80%, and 60%, respectively. Seven out of 50 ccRCCs patient (14%) had histopathological-proven renal sinus invasion. The sensitivity, specificity, PPV, NPV, and accuracy of MDCT in the detection of renal sinus invasion were found 85%, 65%, 28%, 96%, and 68%, respectively. The area under of curve (AUC) value of tumor volume in the detection of perinephric fat invasion was 0.631. The AUC value of tumor volume in the detection of renal sinus invasion was 0.803. CONCLUSION: MDCT has a good sensitivity for detection of renal sinus fat invasion, but low PPV and specificity in patients with ccRCC. Tumor volume, and invasion into the pelvicalyceal structures can aid in the diagnosis of renal sinus fat invasion preoperatively.

Relaciones entre el factor-1alfa inducido por hipoxia y los parámetros de la PET/TC con 18F-FDG en el cáncer colorrectal / Relationships between hypoxia induced factor-1alfa and 18F-FDG PET/CT parameters in colorectal cancer

OBJETIVO: El factor inducible para hipoxia (HIF-1) tiene un papel crítico en la homeostasis del oxígeno y es un activador transcripcional de angiogénesis, eritropoyesis, hierro y metabolismo de glucosa. La tasa de metabolismo de glucosa aumenta en algunos tumores a través de HIF-1alfa. Nuestro objetivo es evaluar la relación entre hipoxia en el cáncer colorrectal, los parámetros de PET, el tamaño del tejido necrótico y los factores pronósticos patológicos mediante el uso de HIF-1alfa. MATERIALES/MÉTODOS: 70 pacientes (28 mujeres/42 hombres; promedio de edad: 63 años) diagnosticados con cáncer colorrectal mediante biopsia, se estadificaron con PET/TC preoperatoria y se operaron posteriormente. La puntuación de evaluación inmunohistoquímica se realizó de acuerdo con la expresión de HIF-1alfa nuclear, la intensidad y la densidad de tinción. El volumen metabólico tumoral (MTV), la glucólisis de lesión total (TLG) y el volumen tumoral (TV) se calculó utilizando el volumen de una fórmula elipsoide mediante las imágenes de TC y el porcentaje de necrosis tumoral (%TmNcr) se calculó por diferencia entre TV y MTV. RESULTADOS: Hubo una correlación positiva moderadamente significativa entre el SUVmáx del tumor y TV y el %TmNcr (r=0,403, p = 0,001 y r=0,5, p = 0,0001, respectivamente). No hubo una relación estadísticamente significativa entre niveles de expresión de HIF-1alfa y SUVmáx tumoral, TLG, MTV, TV, %TmNcr, estadio tumoral, invasión linfovascular, invasión perineural y afectación ganglionar extracapsular/capsular. Por otro lado, se observó una fuerte tinción inmunohistoquímica nuclear en las células tumorales adyacentes al borde invasivo, las células inflamatorias. Aunque no fue estadísticamente significativa, se observó una tinción nuclear moderada o fuerte en el 64,9% de los pacientes metastásicos. CONCLUSIÓN: Aunque la presencia de una correlación positiva entre SUVmáx tumoral y el % de TmNcr muestra que hay células hipóxicas en tejido canceroso con una alta captación de FDG, no se demostró ninguna relación entre la presencia de HIF-1alfa y el incremento metabólico de glucosa y los factores patológicos del tumor. La fuerte tinción inmunohistoquímica nuclear en células tumorales adyacentes a las células inflamatorias y de borde invasivas nos hace pensar que HIF-1alfa desempeña un papel en el área de invasión del microambiente tumoral

AIM: The hypoxia-inducible factor 1 (HIF-1) has a critical role in oxygen homeostasis and it is a transcriptional activator of angiogenesis, erythropoiesis, iron and glucose metabolism. Glucose metabolism rate is increased in some tumours via HIF-1alfa. Our aim is to evaluate the relationship between hypoxia in colorectal cancer, PET parameters, necrotic tissue size and pathologic prognostic factors via using HIF-1alfa. MATERIALS/METHODS: 70 patients (28 female/42 male; median age: 63 years) who were diagnosed with colorectal cancer via biopsy were staged with preoperative PET/CT and operated subsequently. Immunohistochemical evaluation scoring was done according to nuclear HIF-1alfa expression, staining density and intensity. Metabolic tumour volume (MTV), total lesion glycolysis (TLG) and tumour volume (TV) were calculated by using volume of an ellipsoid formula via CT images, and percentage of tumour necrosis (%TmNcr) that was calculated by the difference between TV and recorded MTV. RESULTS: There was a moderately meaningful positive correlation between tumour SUVmax and TV and %TmNcr (r=0.403, p = 0.001 and r=0.500, p = 0.0001, respectively). There were no statistically significant relationships between HIF-1alfa expression levels and tumour SUVmax, TLG, MTV, TV, %TmNcr, tumour stage, lymphovascular invasion, perineural invasion and extracapsular/capsular lymph node involvement. On the other hand, strong nuclear immunohistochemical staining was seen in tumour cells adjacent to invasive border, inflammatory cells. Although not statistically significant, moderate or strong nuclear staining were seen in 64.9% of metastatic patients. CONCLUSION: Although the presence of a positive correlation between tumour SUVmax and %TmNcr shows that there are hypoxic cells in cancer tissue with high FDG uptake, the relationship between the presence of HIF-1alfa and enhanced glucose metabolism and pathological prognostic factors of tumour was not shown. Strong nuclear immunohistochemical staining in tumour cells adjacent to invasive border and inflammatory cells leads us to believe that HIF-1alfa plays a role in the invasion area of tumour microenvironment

Relationships between hypoxia induced factor-1α and 18F-FDG PET/CT parameters in colorectal cancer. / Relaciones entre el factor-1α inducido por hipoxia y los parámetros de la PET/TC con 18F-FDG en el cáncer colorrectal.

AIM: The hypoxia-inducible factor 1 (HIF-1) has a critical role in oxygen homeostasis and it is a transcriptional activator of angiogenesis, erythropoiesis, iron and glucose metabolism. Glucose metabolism rate is increased in some tumours via HIF-1α. Our aim is to evaluate the relationship between hypoxia in colorectal cancer, PET parameters, necrotic tissue size and pathologic prognostic factors via using HIF-1α. MATERIALS/METHODS: 70 patients (28 female/42 male; median age: 63 years) who were diagnosed with colorectal cancer via biopsy were staged with preoperative PET/CT and operated subsequently. Immunohistochemical evaluation scoring was done according to nuclear HIF-1α expression, staining density and intensity. Metabolic tumour volume (MTV), total lesion glycolysis (TLG) and tumour volume (TV) were calculated by using volume of an ellipsoid formula via CT images, and percentage of tumour necrosis (%TmNcr) that was calculated by the difference between TV and recorded MTV. RESULTS: There was a moderately meaningful positive correlation between tumour SUVmax and TV and %TmNcr (r=0.403, p=0.001 and r=0.500, p=0.0001, respectively). There were no statistically significant relationships between HIF-1α expression levels and tumour SUVmax, TLG, MTV, TV, %TmNcr, tumour stage, lymphovascular invasion, perineural invasion and extracapsular/capsular lymph node involvement. On the other hand, strong nuclear immunohistochemical staining was seen in tumour cells adjacent to invasive border, inflammatory cells. Although not statistically significant, moderate or strong nuclear staining were seen in 64.9% of metastatic patients. CONCLUSION: Although the presence of a positive correlation between tumour SUVmax and %TmNcr shows that there are hypoxic cells in cancer tissue with high FDG uptake, the relationship between the presence of HIF-1α and enhanced glucose metabolism and pathological prognostic factors of tumour was not shown. Strong nuclear immunohistochemical staining in tumour cells adjacent to invasive border and inflammatory cells leads us to believe that HIF-1α plays a role in the invasion area of tumour microenvironment.

Atomic resolution force imaging through the static deflection of the cantilever in simultaneous Scanning Tunneling/Atomic Force Microscopy.

We report simultaneous Force -static deflection of the cantilever-, Force Gradient and Scanning Tunneling topography images of Si(111)(7 × 7) surface using an off-resonance small amplitude non-contact atomic force microscopy technique with improved force sensitivity. The signal-to-noise ratio of the fiber interferometer used to detect the deflections of the cantilever was improved by applying an RF-modulation into the diode laser, which suppresses the noise in the laser. The measured sensitivity of ∼20 fm/√Hz allows us to obtain atom resolved images of the surface in static deflection of the cantilever, simultaneously with the other imaging channels.

Investigation of CVD graphene as-grown on Cu foil using simultaneous scanning tunneling/atomic force microscopy.

Scanning tunneling microscopy (STM) and atomic force microscopy (AFM) images of graphene reveal either a triangular or honeycomb pattern at the atomic scale depending on the imaging parameters. The triangular patterns at the atomic scale are particularly difficult to interpret, as the maxima in the images could be every other carbon atom in the six-fold hexagonal array or even a hollow site. Carbon sites exhibit an inequivalent electronic structure in HOPG or multilayer graphene due to the presence of a carbon atom or a hollow site underneath. In this work, we report small-amplitude, simultaneous STM/AFM imaging using a metallic (tungsten) tip, of the graphene surface as-grown by chemical vapor deposition (CVD) on Cu foils. Truly simultaneous operation is possible only with the use of small oscillation amplitudes. Under a typical STM imaging regime the force interaction is found to be repulsive. Force-distance spectroscopy revealed a maximum attractive force of about 7 nN between the tip and carbon/hollow sites. We obtained different contrast between force and STM topography images for atomic features. A honeycomb pattern showing all six carbon atoms is revealed in AFM images. In one contrast type, simultaneously acquired STM topography revealed hollow sites to be brighter. In another, a triangular array with maxima located in between the two carbon atoms was acquired in STM topography.

Everolimus as an mTOR Inhibitor Suppresses Endometriotic Implants: an Experimental Rat Study.

Introduction Mammalian target of rapamycin is a pathway to block apoptosis. Recent studies showed that the activity of mammalian target of rapamycin pathway increases in endometriotic lesions. Aim of the present study was to study the effect of everolimus agent, a rapamycin analog, in an experimental endometriosis model. Materials and Methods Endometriosis established by the autotransplantation of uterine tissue in the peritoneal cavity was confirmed in 24 rats. The animals were then randomly divided into three groups to receive either everolimus (1.5 mg/kg/day, p. o.), anastrozole (0.004 mg/day, p. o.), or normal saline (0.1 mL, i. p.) for 14 days. Endometriotic foci were excised, stained with hematoxylin and eosin, and endometriosis was scored semiquantitatively. In addition, immunohistochemical examination were performed using primary antibodies of vascular endothelial growth factor, CD117, and Bax. Results Both anastrozole and everolimus lowered endometriosis scores. Significant decreases in ovarian follicles were observed following anastrozole treatment but not everolimus treatment. Conclusion Through its apoptosis-promoting effect, everolimus suppressed endometriotic foci without negatively affecting ovarian reserve. These findings support the hypothesis that everolimus merits further study on the way to developing a new endometriosis drug.

Radiation pressure excitation of a low temperature atomic force/magnetic force microscope for imaging in 4-300 K temperature range.

We describe a novel radiation pressure based cantilever excitation method for imaging in dynamic mode atomic force microscopy (AFM) for the first time. Piezo-excitation is the most common method for cantilever excitation, however it may cause spurious resonance peaks. Therefore, the direct excitation of the cantilever plays a crucial role in AFM imaging. A fiber optic interferometer with a 1310 nm laser was used both for the excitation of the cantilever at the resonance and the deflection measurement of the cantilever in a commercial low temperature atomic force microscope/magnetic force microscope (AFM/MFM) from NanoMagnetics Instruments. The laser power was modulated at the cantilever's resonance frequency by a digital Phase Locked Loop (PLL). The laser beam is typically modulated by ∼500 µW, and ∼141.8 nmpp oscillation amplitude is obtained in moderate vacuum levels between 4 and 300 K. We have demonstrated the performance of the radiation pressure excitation in AFM/MFM by imaging atomic steps in graphite, magnetic domains in CoPt multilayers between 4 and 300 K and Abrikosov vortex lattice in BSCCO(2212) single crystal at 4 K for the first time.

Can platelet-rich plasma be used safely in intra-abdominal operations?

OBJECTIVES: In this study, it was aimed to investigate whether or not platelet­rich plasma (PRP) causes intra-abdominal adhesions and therefore, whether or not PRP can be used safely in intra-abdominal operations. METHODS: Of the total of 35 animals, 5 were used as donors for the preparation of platelet­rich plasma (PRP). The surgical procedures were performed on the remaining 30 animals. These rats were randomized and divided into 3 groups of 10. In Group 1, no adhesion induction was performed. Adhesion was induced by cecal abrasion and peritoneal resection model in Groups II and IIII. In Group 2, no treatment was given. In Group 3, 1 cc PRP was applied on the cecum. The rats were sacrificed on postoperative day 21. RESULTS: According to adhesion scores, the difference between the sham and PRP groups was not statistically significant. There was also no significant difference between the control and PRP groups, but the adhesion scores in the PRP group was lower than those in the control group. On histopathological evaluation, the difference between the sham and PRP groups was not statistically significant. There was also no significant difference between the control and PRP groups, but the average fibrosis and inflammation scores in the PRP group were lower than those in the control group. CONCLUSION: The results of the present study have demonstrated that PRP neither reduced nor exacerbated postoperative adhesions. Thus, PRP can be used safely in experimental and clinical studies where it will be applied intra-abdominally (Tab. 2, Fig. 3, Ref. 11).

Targeting leukemia stem cells in vivo with antagomiR-126 nanoparticles in acute myeloid leukemia.

Current treatments for acute myeloid leukemia (AML) are designed to target rapidly dividing blast populations with limited success in eradicating the functionally distinct leukemia stem cell (LSC) population, which is postulated to be responsible for disease resistance and relapse. We have previously reported high miR-126 expression levels to be associated with a LSC-gene expression profile. Therefore, we hypothesized that miR-126 contributes to 'stemness' and is a viable target for eliminating the LSC in AML. Here we first validate the clinical relevance of miR-126 expression in AML by showing that higher expression of this microRNA (miR) is associated with worse outcome in a large cohort of older (⩾60 years) cytogenetically normal AML patients treated with conventional chemotherapy. We then show that miR-126 overexpression characterizes AML LSC-enriched cell subpopulations and contributes to LSC long-term maintenance and self-renewal. Finally, we demonstrate the feasibility of therapeutic targeting of miR-126 in LSCs with novel targeting nanoparticles containing antagomiR-126 resulting in in vivo reduction of LSCs likely by depletion of the quiescent cell subpopulation. Our findings suggest that by targeting a single miR, that is, miR-126, it is possible to interfere with LSC activity, thereby opening potentially novel therapeutic approaches to treat AML patients.

Effects of Ankaferd Blood Stopper and calcium alginate in experimental model of hepatic parenchymal bleeding.

BACKGROUND: This experimental study compared the hemostatic effects of calcium alginate and Anka-ferd Blood Stopper in hepatic parenchymal bleedings. MATERIAL AND METHOD: The study comprised 39 male Wistar albino rats (weight 230±30 g). Laceration model was created in the left lateral lobe of the liver. Standard cotton gauze that was impregnated 0.9% NaCl solution and Calcium alginate cover was compared to ABS tampon. The amount of preoperative bleeding, preoperative and postoperative Day 1 hematocrit levels, and the difference between them were assessed and statistically analyzed. RESULTS: Comparing the hematocrit levels between the groups, we found that the amount of bleeding was significantly higher in the control group versus the study groups (p<0.001). Histopathological examination revealed the portal area enlargement and biliary canaliculi proliferation. In the Ca2+ Alginate group, it was observed that the fibres were still present in the incision line with massive fibrotic area around. In the Ankaferd group, examination of the preparations revealed patchy focal necrosis areas but no fibrotic area. CONCLUSION: With this study, we demonstrated that both calcium alginate and Ankaferd have hemostatic effect in preventing hepatic parenchymal bleeding and that calcium alginate causes fibrosis in the liver, where ABS causes focal necrosis areas(Tab. 2, Fig. 4, Ref. 19).

TDS exposure project: application of the analytic hierarchy process for the prioritization of substances to be analyzed in a total diet study.

The objective of this article is to develop a general method based on the analytic hierarchy process (AHP) methodology to rank the substances to be studied in a Total Diet Studies (TDS). This method was tested for different substances and groups of substances (N = 113), for which the TDS approach has been considered relevant. This work was performed by a group of 7 experts from different European countries representing their institutes, which are involved in the TDS EXPOSURE project. The AHP methodology is based on a score system taking into account experts' judgments quantified assigning comparative scores to the different identified issues. Hence, the 10 substances of highest interest in the framework of a TDS are trace elements (methylmercury, cadmium, inorganic arsenic, lead, aluminum, inorganic mercury), dioxins, furans and polychlorinated biphenyls (PCBs), and some additives (sulfites and nitrites). The priority list depends on both the national situation (geographical variations, consumer concern, etc.) and the availability of data. Thus, the list depends on the objectives of the TDS and on reachable analytical performances. Moreover, such a list is highly variable with time and new data (e.g. social context, vulnerable population groups, emerging substances, new toxicological data or health-based guidance values).

Clinicopathological determinants of positron emission tomography computed tomography fluorodeoxyglucose standardised uptake value in head and neck carcinoma.

BACKGROUND: Although positron emission tomography computed tomography has proven diagnostic and staging value in head and neck carcinoma, it does not have optimal sensitivity or specificity. The positron emission tomography computed tomography fluorodeoxyglucose standardised uptake value has been shown to be associated with carcinoma stage. This study evaluated the impact of major clinicopathological factors on the standardised uptake value at the primary site and at neck lymph node metastases. SUBJECTS AND METHODS: Two hundred and forty-three oral cavity and laryngopharyngeal carcinoma patients who underwent positron emission tomography computed tomography were included. Correlation between the positron emission tomography computed tomography standardised uptake value and various clinicopathological factors was analysed. RESULTS: A positive correlation was found between the standardised uptake value and the size and depth of tumour infiltration, and lymph node positivity. Higher standardised uptake values were seen for more advanced tumour stages. The presence of perineural invasion, lymphatic invasion and extracapsular spread were all associated with increased standardised uptake values. CONCLUSION: Most of the clinicopathological features of head and neck carcinoma which are well known to be poor prognostic factors have a significant impact on positron emission tomography computed tomography fluorodeoxyglucose standardised uptake value.

Antitumor effects of Origanum acutidens extracts on human breast cancer.

PURPOSE: There has been a long-standing interest in the identification of medicinal plants and derived natural products for developing anticancer agents. This work aimed at investigating the antiprolipherative properties of Origanum acutidens (OA) on breast cancer. METHODS: OA water extracts were studied for cytotoxicity against the breast cancer cell lines MCF-7, MDA-MB-468 and MDA-MB-231. In vitro apoptosis studies of these cancer cell lines were performed by annexin V staining in flow cytometry analyses. Immunohistochemistry studies for Ki-67 and caspase-7 of tumor tissue sections of dimethylbenzanthracene (DMBA) -induced mammary cancer in rats were also performed. TUNEL assay was used to detect apoptotic cells of tumor tissue. In vivo anticancer activity testing was carried out by inhibiting the growth of DMBA-induced mammary cancer in rats. RESULTS: OA showed cytotoxicity on all 3 cancer cell lines. Annexin-positive cells level in OA-treated cell lines were significantly higher compared with untreated control cells (p=0.002). The expressions of caspase-7 protein and TUNEL-positive cells were much higher for the rats treated by OA, compared with the untreated control group (p<0.05). The expressions of the Ki-67 decreased in the treated groups compared with the control group (p<0.05). In vivo studies showed that the mean tumor volume inhibition ratio in OA-treated group was 41 % compared with the untreated rats (p<0.05). CONCLUSION: These results indicate that OA has antitumor activity against breast cancer cell lines.

Effect of ozone therapy on autogenous bone graft healing in calvarial defects: a histologic and histometric study in rats.

AIM: The purpose of this study was to analyze histologically the effect of ozone therapy in combination with autogenous bone graft on bone healing in rat calvaria. METHODS: Critical size defects were created in calvaria of 27 male Wistar rats. The animals were divided into three groups of nine animals each: autogenous bone graft group (n = 9); autogenous bone graft with ozone therapy group (80%, 30 s 3 d for 2 wk, n = 9); non-treatment (control) group (n = 9). Animals were killed after 8 wk. Histomorphometric assessments, using image analysis software, and histological analyses were performed. Primary outcome was total bone area. Secondary outcomes (osteoblast number, new bone formation) were also measured. RESULTS: Histomorphometrically, the total bone area in the autogenous bone graft with ozone therapy group (9.3 ± 2.2) were significantly higher than that of the autogenous bone graft group (5.1 ± 1.8) (p < 0.05). Also, the ozone therapy group significantly increased the percentage of total bone area compared to the autogenous bone graft group (p < 0.05). The osteoblast number significantly increased in the autogenous bone graft with the ozone therapy group (58 ± 12.3) compared to the autogenous bone graft group (9.3 ± 3.5) (p < 0.05). Also, it was observed that autogenous bone graft with ozone therapy group showed significant new bone formation when compared to the autogenous bone graft group (p < 0.05). CONCLUSION: Ozone therapy enhances new bone formation by autogenous bone graft in the rat calvarial defect model.