Otitis media with effusion (OME) is an inflammation of the middle ear in which a collection of liquid is present in the middle-ear space while the tympanic membrane is intact. The association between adenoid inflammation and OME has long been noted but the exact mechanism is still much debated. We studied the role of adenoid mast cells in the causation of OME.To study the distribution and role of adenoid mast cells in the causation of OME.A cross-sectional, prospective study was carried out in the otorhinolaryngologic clinic, department of otorhinolaryngology (ORL), Science University of Malaysia, from June 1999 to September 2001. A total number of 50 cases were studied. Twenty-five of these patients underwent adenoidectomy, while another 25 patients underwent adenoidectomy and myringotomy with ventilation tube insertion. The adenoid specimens from all patients were examined for the number of adenoid mast cells present, using light microscopy and toluidine blue as the staining agent. The results were analysed using SPSS version 10.0 computer software.The population of adenoid mast cells in children with OME was significantly greater than that in children without OME (p=0.000).The increased number of adenoid mast cells in patients with OME suggests that inflammation may play a role in this condition.
Background: Rapamycin is an effective anti-angiogenic drug. However, the mode of its action remains unclear. Therefore, in this study, we aimed to elucidate the antitumor mechanism of rapamycin, hypothetically via apoptotic promotion, using MCF-7 breast cancer cells. Materials and Methods: MCF-7 cells were plated at a density of $1{\times}10^5$ cells/well in 6-well plates. After 24h, cells were treated with a series of concentrations of rapamycin while only adding DMEM medium with PEG for the control regiment and grown at $37^{\circ}C$, 5% $CO_2$ and 95% air for 72h. Trypan blue was used to determine the cell viability and proliferation. Untreated and rapamycin-treated MCF-7 cells were also examined for morphological changes with an inverted-phase contrast microscope. Alteration in cell morphology was ascertained, along with a stage in the cell cycle and proliferation. In addition, cytotoxicity testing was performed using normal mouse breast mammary pads. Results: Our results clearly showed that rapamycin exhibited inhibitory activity on MCF-7 cell lines. The $IC_{50}$ value of rapamycin on the MCF-7 cells was determined as $0.4{\mu}g/ml$ (p<0.05). Direct observation by inverted microscopy demonstrated that the MCF-7 cells treated with rapamycin showed characteristic features of apoptosis including cell shrinkage, vascularization and autophagy. Cells underwent early apoptosis up to 24% after 72h. Analysis of the cell cycle showed an increase in the G0G1 phase cell population and a corresponding decrease in the S and G2M phase populations, from 81.5% to 91.3% and 17.3% to 7.9%, respectively. Conclusions: This study demonstrated that rapamycin may potentially act as an anti-cancer agent via the inhibition of growth with some morphological changes of the MCF-7 cancer cells, arrest cell cycle progression at G0/G1 phase and induction of apoptosis in late stage of apoptosis. Further studies are needed to further characterize the mode of action of rapamycin as an anti-cancer agent.
Stain normalization is an image pre-processing method extensively used to standardize multiple variances of staining intensity in histopathology image analysis. Staining variations may occur for several reasons, such as unstandardized protocols while preparing the specimens, using dyes from different manufacturers, and varying parameters set while capturing the digital images. In this study, a double stain normalization technique based on immunohistochemical staining is developed to improve the performance of the conventional Reinhard’s algorithm. The proposed approach began with preparing a target image by applying the contrast-limited adaptive histogram equalization (CLAHE) technique to the targeted cells. Later, the colour distribution of the input image will be matched to the colour distribution of the target image through the linear transformation process. In this study, the power-law transformation was applied to address the over-enhancement and contrast degradation issues in the conventional method. Five quality metrics comprised of entropy, tenengrad criterion (TEN), mean square error (MSE), structural similarity index (SSIM) and correlation coefficient were used to measure the performance of the proposed system. The experimental results demonstrate that the proposed method outperformed all conventional techniques. The proposed method achieved the highest average values of 5.59, 3854.11 and 94.65 for entropy, TEN, and MSE analyses.
Detecting and classifying the Ki67 expression is a challenging process. The inconsistency in staining intensity and the variations in image quality are the main factors that may reduce the performance of an automated system. Therefore, this study proposes a framework that objectively improves detecting and classifying Ki67 expression in astrocytoma histopathological images. The proposed framework began with implementing the double stain normalization procedure to reduce the colour-staining intensity variations. Then, the system automatically enhanced the morphological features of the Ki67 expression. The following step was to segment the enhanced images by using the U-Net network model. The last step of the proposed framework was to localize and classify the Ki67 expression based on the modified YOLOv3 model. In conclusion, the proposed YOLOv3 model produced a high detection result with a mean average precision of 0.80 for detecting Ki67-positive cells and 0.87 for detecting Ki67-negative cells.
Objective: To analyze the effect of sirolimus and sunitinib in blocking the tumor growth and to evaluate the expressions of estrogen receptor (ER), progesterone receptor (PgR), and human epidermal growth factor receptor-2 (HER2/neu) after treated with sirolimus and sunitinib. Methods: Thirty-two female Sprague Dawley rats at age 21-days old were administered intraperitoneally with N-Methyl-N-Nitroso Urea (NMU), dosed at 70mg/kg body weight. The rats were divided into 4 groups; Group 1 (Control, n=8), Group 2 (Sirolimus, n=8), Group 3 (Sunitinib, n=8) and Group 4 (Sirolimus+Sunitinib, n=8), being treated twice when the tumor reached the size of 14.5±0.5 mm and subsequently sacrificed after 5 days. The protein expressions of ER, PgR and HER2/neu of the tumor tissues were evaluated by using immunohistochemistry analysis. Results: Treatment with sirolimus alone lowered expressions of ER and PgR of breast cancer and reduced tumor size. There was no significant difference of ER and PgR expressions between control and sunitinib treated tumor. Sunitinib treated tumors reduce in diameter after the first treatment, however the diameter increases after the second treatment. Histologically, sunitinib treated tumor did not show any aggressive invasive carcinoma of no special type (NST) histological subtypes. In addition, all NMU-induced tumors are HER2/neu-negative scoring. Conclusion: Sirolimus is neither synergistic nor additive with sunitinib for breast cancer treatment.
Paired-like homeodomain transcription factor 2 (PITX2) is another new marker in breast carcinoma since hypermethylation at P2 promoter of this gene was noted to be associated with poor prognosis. We investigated the expression of PITX2 protein using immunohistochemistry in invasive ductal carcinoma and its association with the established growth receptors such as estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth receptor 2 (HER2).We conducted a cross sectional study using 100 samples of archived formalin-fixed paraffin embedded tissue blocks of invasive ductal carcinoma and stained them with immunohistochemistry for PITX2, ER, PR and HER2. All HER2 with scoring of 2+ were confirmed with chromogenic in-situ hybridization (CISH).PITX2 protein was expressed in 53% of invasive ductal carcinoma and lack of PITX2 expression in 47%. Univariate analysis revealed a significant association between PITX2 expression with PR (p=0.001), ER (p=0.006), gland formation (p=0.044) and marginal association with molecular subtypes of breast carcinoma (p=0.051). Combined ER and PR expression with PITX2 was also significantly associated (p=0.003) especially in double positive cases. Multivariate analysis showed the most significant association between PITX2 and PR (RR 4.105, 95% CI 1.765-9.547, p=0.001).PITX2 is another potential prognostic marker in breast carcinoma adding significant information to established prognostic factors of ER and PR. The expression of PITX2 together with PR may carry a very good prognosis.
Eighteen male Wistar rats aged six weeks were divided equally into Methamphetamine (MA), Placebo and Control group. MA group were injected with 50mg/kg body weight of Methamphetamine hydrochloride (MAHCl) in normal saline, Placebo group were injected with normal saline only, while Control group not injected with anything. Five MA group rats died within four hours of injection and their hearts collected on the same day. Another MA group rat was sacrificed two days after injection. Placebo and control group were sacrificed at similar intervals. Collected hearts were studied for cardiac lesions under light microscopy using special staining and immunohistochemistry. Microscopic examination of the myocardium of the rats that died on the first day of injection showed loss of nuclei in some myocytes, indicating cell death. Some areas in the sub-endocardium region showed internalization and enlargement of myocyte nuclei, consistent with regeneration of cells. There were very few foci of necrosis observed in these samples. The heart samples from the single rat that survived injection for two days showed foci of infiltration of macrophage-like cells that were later revealed to be regenerating myocytes. There were also spindle-like fibroblasts, macrophages and a few leucocytes found within these foci. The overall appearance of the myocardium did not indicate any inflammatory response, and the expected signs of necrosis were not observed. These results suggest a need to re-evaluate the toxic and lethal dosages of MA for use in animals testing. Cause of death was suspected to be due to failure of other major organs from acute administration of MA. Death occurred within a time period where significant changes due to necrosis may not be evident in the myocardium. Further investigations of other organs are necessary to help detect death due to acute dosage of MA.
Image segmentation is a key step in most medical image analysis. However, the process is particularly difficult due to limitation of the imaging equipments and variation in biological system. Therefore, accurate and robust segmentation are important for quantitative assessment of medical images in order to achieve correct clinical diagnosis. This paper studies the performance of clustering and adaptive thresholding algorithms for segmenting the tuberculosis (TB) bacilli in tissue sections. Images are obtained by analyzing the Ziehl-Neelsen (ZN) stained tissue slide and capturing using a digital camera attached to a light microscope. Three clustering algorithms namely k-mean clustering, moving k-mean clustering and fuzzy c-mean clustering, and two adaptive thresholding algorithms, Otsu and iterative thresholding, are evaluated for segmentation of TB bacilli. The saturation component, derived from C-Y colour model is utilised as input to these algorithms as it provides good separation between the TB bacilli and the background. The segmentation results are further compared with the manual-segmentation image. Acceptable segmentation accuracy of up to 99.00% was achieved by using the clustering algorithms and the Otsu's thresholding. However, k-mean clustering was chosen as it produced the highest TB segmentation rate.
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