MMP-9 and CCR7 as Possible Predictors of Lymph Node Metastasis in Laryngeal Squamous Cell Carcinoma.

Background & Objective: The expression of matrix metalloproteinase-9 (MMP-9) and chemokine receptor 7 (CCR7) is significantly associated with tumor invasion and metastasis. Little is known regarding the potential of these markers in predicting cancer metastasis in Laryngeal Squamous Cell Carcinoma (LSCC). Therefore, this study aimed to dissect the potential of these markers in predicting the lymph node metastasis in LSCC patients. Methods: Sixty tissue samples were obtained from the patients diagnosed pathologically with LSCC who underwent partial or total laryngectomy. The expression of MMP-9 and CCR7 was measured using the immunohistochemistry staining in the tissue samples of LSCC patients. The ROC (receiver operating characteristic) curve was used to determine the most significant cut-off points of expression according to the highest sensitivity and specificity of both the markers to predict the lymph node metastasis in LSCC. Then, the relationship between the clinicopathology features and the expression of MMP-9 and CCR7 was evaluated. Results: The expression of both MMP-9 and CCR7 was significantly correlated with the lymph node metastasis in LSCC (P<0.001). Furthermore, CCR7 expression exhibited the highest prediction accuracy (AUC 95.7%) and sensitivity (100%) in predicting the lymph node metastasis in LSCC compared to that of MMP-9 (AUC 92.9%, sensitivity 90%). We also found that patients with larger tumor size (> 4 cm) had significantly higher expression of MMP-9 and CCR7 (P<0.002 and P<0.001, respectively). The Elevated expression level of CCR7 statistically correlated with higher MMP-9 expression (P<0.001). Conclusion: MMP-9 and CCR7 might be beneficial as predictors of lymph node metastasis in LSCC patients.

Accuracy Analysis of Deep Learning Methods in Breast Cancer Classification: A Structured Review.

Breast cancer is diagnosed using histopathological imaging. This task is extremely time-consuming due to high image complexity and volume. However, it is important to facilitate the early detection of breast cancer for medical intervention. Deep learning (DL) has become popular in medical imaging solutions and has demonstrated various levels of performance in diagnosing cancerous images. Nonetheless, achieving high precision while minimizing overfitting remains a significant challenge for classification solutions. The handling of imbalanced data and incorrect labeling is a further concern. Additional methods, such as pre-processing, ensemble, and normalization techniques, have been established to enhance image characteristics. These methods could influence classification solutions and be used to overcome overfitting and data balancing issues. Hence, developing a more sophisticated DL variant could improve classification accuracy while reducing overfitting. Technological advancements in DL have fueled automated breast cancer diagnosis growth in recent years. This paper reviewed studies on the capability of DL to classify histopathological breast cancer images, as the objective of this study was to systematically review and analyze current research on the classification of histopathological images. Additionally, literature from the Scopus and Web of Science (WOS) indexes was reviewed. This study assessed recent approaches for histopathological breast cancer image classification in DL applications for papers published up until November 2022. The findings of this study suggest that DL methods, especially convolution neural networks and their hybrids, are the most cutting-edge approaches currently in use. To find a new technique, it is necessary first to survey the landscape of existing DL approaches and their hybrid methods to conduct comparisons and case studies.

Efficacy of Using Bacillus subtilis Enzyme as a Caffeine Level Reducer in Cascara Robusta Coffee (Coffea canephora L.).

<b>Background and Objective:</b> An increase in the consumption of robusta coffee resulted in an increase in waste from coffee, one of the coffee wastes, namely coffee bean skins or cascara. Robusta coffee cascara contains 1-1.3 g of caffeine which causes side effects, such as insomnia and seizures etc. So this research aims to reduce the caffeine content in cascara by using <i>Bacillus subtilis</i>. Using optimum conditions and capabilities. <b>Materials and Methods:</b> The experiment was conducted from May to August, 2022 in the Pharmacy Laboratory, Faculty of Mathematics and Natural Sciences, Universitas Pakuan, Indonesia. Before optimizing, cascara was extracted using the ultrasonic assisted extraction (UAE) method, validated by the High-Performance Liquid Chromatography (HPLC) method to determine caffeine content and a paired sample t-test was performed using Statistical Package for the Social Sciences (SPSS). <b>Results:</b> It showed that in validating the HPLC method, the wavelength of caffeine in cascara was 272 nm. The mobile phase was a mixture of methanol-water (adjust orthophosphate). The pH (2.4) (45:55), obtained the optimum decaffeination conditions at the concentration of bacteria <i>Bacillus subtilis</i> 6% and a long incubation time of 24 hrs resulted in a decrease in caffeine content of 51.3843±0.2503%. <b>Conclusion:</b> The results of the paired sample t-test indicate that the concentration of bacteria <i>Bacillus subtilis</i> and incubation time significantly influence caffeine levels.

Ethanol Extract of Pomegranate (Punica granatum) Peel in Increasing the Expression of Caspase-3 in DSS-Induced Mice.

BACKGROUND: Colorectal cancer (CRC) is a malignancy derived from the glandular epithelial cells in the colon. Patients with inflammatory bowel disease (IBD) are more likely to develop CRC. Cancer proliferation is characterized by the loss of inhibition of apoptosis, which involves caspase-3 activation. This study examined the effects of the pomegranate peel extract on the expression of caspase-3 in mice crypt cells induced by dextran sodium sulfate (DSS) 2%. METHODS: The experimental study was done in six groups. All treatments were done in 42 days. The groups were all induced by DSS through water drinking, except for the normal group, which was only given water. The treatments given included the pomegranate extract in two doses (240 mg and 480 mg/kg bw/day), aspirin, and ellagic acid. The specimens were then fixated and stained for the immunohistochemistry scoring for the expression of caspase-3, which was then analyzed statistically. RESULTS: The H-scores of each treatment group were 213.23 ± 8.32 (DSS group), 243.81 ± 18.69 (normal group), 226.10 ± 12.38 (pomegranate peel extract of 240 mg/kg/d), 238.84 ± 15.81 (pomegranate peel extract of 480 mg/kg/d), 227.47 ± 12.15 (aspirin), and 224.01 ± 18.39 (ellagic acid). Statistical differences were found in one-way analysis of variance (ANOVA) and post hoc analysis among the DSS group, normal group, and dose 2 group (pomegranate peel extract of 480 mg/kg/day). CONCLUSIONS: The ethanol extract of pomegranate was able to induce apoptosis, which was demonstrated by the increase of caspase-3 expression.

Correlation between CD 34 and CD 68 expression in placental malaria with maternal anemia.

BACKGROUND: Malaria is the second most life-threatening infectious disease in Indonesia, causing approximately 1-3 million deaths annually. Histopathologic studies assessing CD 68 and CD 34 protein expression in placental malaria and its association with maternal anemia are essential to determine the prognosis of malaria in pregnancy. MATERIALS AND METHODS: This cross-sectional study was carried out in 2017. Thirty biopsy samples of human placental tissue were obtained from Timika and Sumba, and ten normal biopsy samples were taken from the Pathological Anatomy Department of Cipto Mangunkusumo General Hospital as comparisons. CD 34 and CD 68 protein expressions were determined using immunohistochemistry, and the resulting data were analyzed using SPSS. RESULTS: Average hemoglobin (Hb) level was 9.5 mg/dL, 11.5 mg/dL, and 9.9 mg/dL in acute infection, chronic infection, and latent infection, respectively. A positive correlation was found between CD 68 protein expression and maternal Hb level. No correlation was found between CD34 expression and maternal anemia. CONCLUSIONS: CD 68 expression in placental tissue biopsy from Timika and Sumba residents with placental malaria was shown to be positively correlated with maternal anemia. Immunohistochemical examination of CD 68 may play a role in the early diagnosis of malaria.

The potential of lunasin extract for the prevention of breast cancer progression by upregulating E-Cadherin and inhibiting ICAM-1.

Background: Research in natural substances for their anticancer potential has become increasingly popular. Lunasin, a soybean protein, is known to inhibit cancer progression via various pathways.  The aim of this study was to investigate the effect of Lunasin Extract (LE) on the expression of Intercellular Adhesion Molecule 1 (ICAM-1) and epithelial cadherins (E-Cadherin) in breast cancer. Methods: In this true-experimental in vivo study, 24 Sprague-Dawley rats that were induced by 7,12-Dimethylbenz[a]anthracene (DMBA), were used. Based on the therapy given, the groups were divided into, normal, positive control (PC), negative control (NC), adjuvant, curative, and preventive. Lunasin was extracted from soybean seeds of the Grobogan variety in Indonesia. Tissue samples were obtained, processed, stained with anti-ICAM-1 and anti-E-Cadherin antibodies, examined under a microscope, and quantified using H-score. The data were analyzed using ANOVA, which was then followed by Duncan's test.  Results: Statistically significant difference in ICAM-1 expression was observed between the following groups: adjuvant and NC, normal and NC, PC and NC, adjuvant and preventive, normal and preventive, PC and preventive, adjuvant and curative, normal and curative, PC and curative. E-Cadherin expression was significantly different between preventive and NC, adjuvant and NC, PC and NC, normal and NC, adjuvant and curative, PC and curative, normal and curative, normal and preventive. Significant negative correlation was found between ICAM-1 and E-Cadherin [-0.616 (-0.8165; -0.283)] with p = 0.001.  Conclusion: Preventive dose of LE was able to reduce ICAM-1 expression while increasing E-Cadherin expression.

Xanthine Oxidase-Induced Inflammatory Responses in Respiratory Epithelial Cells: A Review in Immunopathology of COVID-19.

Xanthine oxidase (XO) is an enzyme that catalyzes the production of uric acid and superoxide radicals from purine bases: hypoxanthine and xanthine and is also expressed in respiratory epithelial cells. Uric acid, which is also considered a danger associated molecule pattern (DAMP), could trigger a series of inflammatory responses by activating the inflammasome complex path and NF-κB within the endothelial cells and by inducing proinflammatory cytokine release. Concurrently, XO also converts the superoxide radicals into hydroxyl radicals that further induce inflammatory responses. These conditions will ultimately sum up a hyperinflammation condition commonly dubbed as cytokine storm syndrome (CSS). The expression of proinflammatory cytokines and neutrophil chemokines may be reduced by XO inhibitor, as observed in human respiratory syncytial virus (HRSV)-infected A549 cells. Our review emphasizes that XO may have an essential role as an anti-inflammation therapy for respiratory viral infection, including coronavirus disease 2019 (COVID-19).

Potential of AKT2 expression as a predictor of lymph-node metastasis in invasive breast carcinoma of no special type.

BACKGROUND: Invasive breast carcinoma of no special type (IBC-NST) is the most common type of breast cancer and mainly causes regional lymph-node metastasis (LNM). We investigated the potential for AKT2 expression as a predictive biomarker for LNM in IBC-NST. METHODS: Forty-eight paraffin blocks containing IBC-NST primary tumors were divided into two groups based on presence or absence of LNM. Age, tumor grade, tumor size, lymphovascular invasion (LVI), and AKT expression were assessed. AKT2 expression was assessed based on immunohistochemical staining, while other data were collected from archives. RESULTS: Multiple logistic regression results showed that AKT2 expression and LVI were significantly associated with LNM (odds ratio [OR], 5.32; 95% confidence interval [CI], 1.42 to 19.93 and OR, 4.46; 95% CI, 1.17 to 16.97, respectively). AKT2 expression was able to discriminate against LNM (area under the receiver operating characteristic, 0.799 ± 0.063; 95% CI, 0.676 to 0.921) at an H-score cutoff of 104.62 (83.3% sensitivity, 62.5% specificity). CONCLUSIONS: AKT2 expression has potential as a predictor of LNM in IBC-NST. The H-score cutoff for AKT2 expression can be used as a classification guide in future studies.

Histopathology and ARID1A Expression in Endometriosis- Associated Ovarian Carcinoma (EAOC) Carcinogenesis Model with Endometrial Autoimplantation and DMBA Induction.

BACKGROUND: Ovarian carcinoma is one of the most deadly malignancies in the gynecologic field. The cause is not yet known, and the clinical symptoms are not specific. Endometrioid carcinoma and ovarian clear cell carcinoma can originate from endometriosis and are known as endometriosis-related ovarian carcinoma (EAOC). Development of EAOC experimental animal models is needed for basic research and clinical preparation of human tissue tests. This study aimed to determine the role of the ARID1A gene mutation in the carcinogenetic process of EAOC in experimental animal models induced with DMBA. METHODS: In this study, the EAOC experimental model was developed using the autoimplantation technique and DMBA induction. This study involved placebo surgery mice (sham), endometrial autoimplantation, and a combination of endometrial autoimplantation and DMBA induction, which were sacrificed at weeks 5, 10, and 20, respectively. Histopathological assessment and immunohistochemical ARID1A staining with an assessment of positive percentages were carried out on 200 cells. RESULTS: This study produced 1 (20%) atypical endometriosis and 1 (20%) clear cell carcinoma at implantation and after 10 weeks of DMBA induction, and 100% endometrioid carcinoma in the DMBA-induced group. ARID1A staining did not show any significant difference (p = 0.313) in all groups. CONCLUSION: The combination of endometrial autoimplantation techniques and DMBA induction in the ovary produced atypical endometriosis, clear cell carcinoma, and endometrioid carcinoma, where time is an important factor. There was no significant difference in ARID1A expression between the treatment and control groups.

Potential of AKT2 expression as a predictor of lymph-node metastasis in invasive breast carcinoma of no special type

Background@#Invasive breast carcinoma of no special type (IBC-NST) is the most common type of breast cancer and mainly causes regional lymph-node metastasis (LNM). We investigated the potential for AKT2 expression as a predictive biomarker for LNM in IBC-NST. @*Methods@#Forty-eight paraffin blocks containing IBC-NST primary tumors were divided into two groups based on presence or absence of LNM. Age, tumor grade, tumor size, lymphovascular invasion (LVI), and AKT expression were assessed. AKT2 expression was assessed based on immunohistochemical staining, while other data were collected from archives. @*Results@#Multiple logistic regression results showed that AKT2 expression and LVI were significantly associated with LNM (odds ratio [OR], 5.32; 95% confidence interval [CI], 1.42 to 19.93 and OR, 4.46; 95% CI, 1.17 to 16.97, respectively). AKT2 expression was able to discriminate against LNM (area under the receiver operating characteristic, 0.799 ± 0.063; 95% CI, 0.676 to 0.921) at an H-score cutoff of 104.62 (83.3% sensitivity, 62.5% specificity). @*Conclusions@#AKT2 expression has potential as a predictor of LNM in IBC-NST. The H-score cutoff for AKT2 expression can be used as a classification guide in future studies.

Potential of AKT2 expression as a predictor of lymph-node metastasis in invasive breast carcinoma of no special type

Background@#Invasive breast carcinoma of no special type (IBC-NST) is the most common type of breast cancer and mainly causes regional lymph-node metastasis (LNM). We investigated the potential for AKT2 expression as a predictive biomarker for LNM in IBC-NST. @*Methods@#Forty-eight paraffin blocks containing IBC-NST primary tumors were divided into two groups based on presence or absence of LNM. Age, tumor grade, tumor size, lymphovascular invasion (LVI), and AKT expression were assessed. AKT2 expression was assessed based on immunohistochemical staining, while other data were collected from archives. @*Results@#Multiple logistic regression results showed that AKT2 expression and LVI were significantly associated with LNM (odds ratio [OR], 5.32; 95% confidence interval [CI], 1.42 to 19.93 and OR, 4.46; 95% CI, 1.17 to 16.97, respectively). AKT2 expression was able to discriminate against LNM (area under the receiver operating characteristic, 0.799 ± 0.063; 95% CI, 0.676 to 0.921) at an H-score cutoff of 104.62 (83.3% sensitivity, 62.5% specificity). @*Conclusions@#AKT2 expression has potential as a predictor of LNM in IBC-NST. The H-score cutoff for AKT2 expression can be used as a classification guide in future studies.

Increased Estradiol Levels Effect on Leptin Expression in Midluteal Endometrial Tissue of Macaca Nemestrina (Southern Pig-Tailed Macaques).

Objective: To measure the effect of serum estradiol (E2) levels on leptin expression in Macaca nemestrina (southern pig-tailed macaque) endometrial tissue. Materials and methods: This study used paraffin-embedded midluteal phase endometrial tissue blocks of Macaca nemestrina from previous study. Included subjects were 15 female macaques of reproductive age (8-10 years) with a previous history of producing offspring, which were divided into four groups: groups administered with 30 IU, 50 IU, and 70 IU r-FSH (intervention group), and no r-FSH (control group). The stimulation was done following GnRH agonist long protocol. Staining was done using immunohistochemistry. Leptin expression was measured using immunohistochemistry (IHC) Profiler plugin of ImageJ software and counted semi-quantitatively as Histological Score (Hscore). Results: Correlation between E2 concentration to stromal leptin expression was observed (p=0.043). Conclusion: Serum estradiol concentration is found to be correlated with leptin expression in Macaca nemestrina, suggesting a mechanism of decreasing endometrial receptivity among women undergoing controlled ovarian hyperstimulation.

Increased Progesterone on the Day of Administration of hCG in Controlled Ovarian Hyperstimulation Affects the Expression of HOXA10 in Primates' Endometrial Receptivity.

The increase in progesterone (P4) levels on the day of human chorionic gonadotropin (hCG) administration have a negative effect on endometrial receptivity. There are few reports regarding the expression of homeobox A10 (HOXA10) as one of many biomolecular factors of endometrial receptivity. To evaluate the effect of increased P4 concentration on the day of hCG administration on HOXA10, a total of 16 Macaca nemestrina were divided into three dose groups of recombinant-follicle stimulating hormone (rFSH) (30IU, 50IU, and 70IU) and one control group. Injection of rFSH combined with gonadotropin release hormone (GnRH) at 160 ug/day was given subcutaneously using a long protocol technique. Blood samples for estradiol (E2) and (P4) concentration measurements were taken on the day of injecting hCG in the final follicular phase, while the collection of endometrial tissue for HOXA10 measurement was carried out 8 to 10 days after hCG administration. E2 and P4 were measured by ELISA, whereas HOXA10 expression was measured with immunohistochemical (IHC) techniques. The concentration of E2 and P4 was found to be higher in dose groups compared with the natural group, but no significant differences were found within the group. For the Hscore for HOXA10 expression, no significant differences within dose groups were found. In addition, no significant differences for the Hscore for HOXA10 were found when compared to E2 groups. Significantly, the Hscore of HOXA10 was found to be >1 ng/mL in the P4 group compared with the Hscore HOXA10 in the P4 natural group (p = 0.022). The high concentration of P4 caused by ovarian hyperstimulation in the follicular phase stimulates the expression of HOXA10 in the secretion phase.

The effect of lunasin from Indonesian soybean extract on histopatologic examination and cox-2 expression in dextran sodium sulfate-induced mice colon.

Inflammatory bowel disease (IBD) is a condition describing chronic gastrointestinal inflammation. Chronic inflammation in colon can develop into colon cancer. Lunasin has been known to inhibit inflammatory reactions induced by lipopolysaccharide in vitro. The effect of lunasin to inhibit inflammation in vivo is not widely known. In this study, we analyzed the effect of lunasin from soybean to decrease the risk of inflammation by analyzing histopathologic feature and the expression of COX-2. 30 mice are divided into 6 groups. Normal group was not induced by dextran sodium sulfate (DSS). The other groups were induced by 2% DSS through drinking water for 9 days. After 9 days, negative control group did not receive any treatment. The other groups received treatment given lunasin dose 20 mg/kg body weight (BW) and 40 mg/kg BW, commercial lunasin and positive control given aspirin. Treatment was performed for 5 weeks. Inflammatory colon histopathologic examination and immunohistochemical score of COX-2 proteins were analyzed using statistical tests. Lunasin dose 20 mg/kg BW and 40 mg/kg BW were able to significantly reduce inflammation (P<0.05) performed by histopathologic feature with an average score of 2.52 and 2.16 COX-2 expression decreased significantly (P<0.05) with an average score of 43.674 and 33.349. Lunasin dose 20 mg/kg BW and 40 mg/kg BW were able to inhibit inflammation and decrease the expression of COX-2 in colon induced by DSS.