TGF-ß1 Effects on Total Collagen of the New Zealand Rabbit's Urethral Wall (Oryctolagus cuniculus) in Animal Models of Urethral Stricture.

Background: Currently, animal models of urethral stricture are not standardized. Transforming Growth Factor Beta 1 (TGF-ß1) regulates extracellular matrix deposition in homeostatic and pathological responses. Objective: The aim of this study was to present the potential model to be developed as a urethral stricture. Methods: True experimental laboratory research was conducted by using Male New Zealand rabbits (Oryctolagus cuniculus), which were divided into 5 groups; control, placebo, and 3 treatment groups (TGF-ß1 injection of 1 µg, 2 µg, 4 µg). Urethrography, histopathological analysis, and evaluation of total collagen formation of the urethral wall were performed after 6 weeks. Results: An increase in the dose of TGF-ß1 decreased the mean rabbit's urethral lumen diameter (29.3% in the 2µg group and 34% in the 4µg group) compared to controls. Three rabbits decreased as much as ≤ 50% in urethral lumen diameter. Significant increases in total collagen density in the periluminal and peripheral urethral spongiosum were noted by increasing doses of TGF-ß1. The percentage of urethral lumen diameter has a strong negative correlation with periluminal total collagen density (r = -0,798; p = 0,000) and very strong negative correlation with peripheral spongiosa total collagen density (r = -0,748, p = 0,000). Conclusion: TGF-ß1 plays a role in changing total collagen compositions of the rabbit's urethral wall, decreasing the urethral lumen diameter. Further research with increasing doses of TGF-ß1 is needed to determine the effective dose of TGF-ß1 in inducing urethral stricture.

Deciphering the Role of TGF-ß1 in Altering Collagen I and Collagen III in the New Zealand Rabbit's (Oryctolagus cuniculus) Urethral Wall in Urethral Stricture Development.

Background: Presently, there's a lack of standardization in animal models used for studying urethral stricture. Transforming Growth Factor Beta 1 (TGF-ß1) is known to regulate the deposition of extracellular matrix in both normal and pathological conditions. This factor holds promise as a potential model for simulating urethral stricture. Objective: This study aims to investigate the impact of Transforming Growth Factor Beta 1 (TGF-ß1) on Collagen I and Collagen III within the urethral wall of New Zealand Rabbits (Oryctolagus cuniculus) in the context of developing urethral stricture in animal models. Methods: We conducted genuine laboratory experiments using Male New Zealand rabbits (Oryctolagus cuniculus), which were categorized into five groups: control, placebo, and three treatment groups (TGF-ß1 injections of 1 µg, 2 µg, 4 µg). After a duration of 6 weeks, we conducted urethrography, histopathological analysis, and assessed the formation of collagen I and collagen III within the urethral wall. Results: Elevating the dosage of TGF-ß1 led to a reduction in the average urethral lumen diameter of rabbits (29.3% in the 2µg group and 34% in the 4µg group) compared to the control group. In fact, three rabbits experienced a decrease of ≤ 50% in their urethral lumen diameter. As the doses of TGF-ß1 increased, we observed significant increases in the density of collagen I, and collagen III in both the periluminal and peripheral regions of the urethral spongiosum. Additionally, there was a tendency for the collagen I/collagen III ratio to decrease in the periluminal region, with collagen III density surpassing that of collagen I. In the peripheral spongiosa area, notable mean differences were observed between the control group, 1T, and 2T groups, with collagen I density tending to be higher than that of collagen III. Furthermore, the percentage of urethral lumen diameter exhibited a robust negative correlation with periluminal collagen I density (r = -0.672, p = 0.001), peripheral spongiosa collagen I density (r = -0.603, p = 0.005), periluminal collagen III density (r = -0.717, p = 0.001), and an exceptionally strong negative correlation with collagen III density of peripheral spongiosa (r = -0.804, p = 0.000). Conclusion: TGF-ß1 exerts an influence on altering the composition of collagen I and collagen III within the urethral wall of rabbits, leading to a reduction in the diameter of the urethral lumen. Further research is warranted to determine the optimal dose of TGF-ß1 required to induce urethral stricture effectively.

The Effect of Long-Term Tamsulosin Monotherapy and Tamsulosin - Dutasteride Combination Therapy on PKC-α Enzyme Expression in Prostate Stromal Tissue.

Background: The α-adrenergic receptor antagonist is the most effective medical therapy to reduce the dynamic component in patients with BPH. However, long-term administration of receptor antagonists can cause upregulation of mRNA receptor expression, resulting in tolerance of drug effectiveness. PKC-α is involved in the process of prostate smooth muscle contraction through activation of the voltage-gated Ca2+ conducted canal, influenced by androgen hormones, especially testosterone, and has an isoform with Twist1, a transcription factor that plays a role in up-regulation of androgen receptors. Objective: The aim of the study was to compare the effect of long-term tamsulosin monotherapy and tamsulosin - dutasteride combination therapy in PKC-α enzyme expression in prostate stromal tissue of Rattus norvegicus rats of Wistar strain. Methods: Out of 80 samples of Rattus norvegicus rats were divided into 8 groups with different interventions: negative control group, positive control group, tamsulosin monotherapy administration for 1 day, 3 day, and 6 day groups, and tamsulosin - dutasteride combination therapy for 1 day, 3 day, and 6 day groups. BPH was induced with 3 mg/kg of testosterone proprionate for 3 weeks, continued with drugs administration according to intervention grouping. Prostate stromal tissue was taken and prepared for PKC-α enzyme measurement with ELISA method. Results: There was a significant difference (p<0.05) in the effect of tamsulosin monotherapy and tamsulosin-dutasteride combination therapy on the PKC-α expression. There was a strong positive relationship between the duration of tamsulosin-dutasteride combination therapy on the PKC-α expression, which means the longer the duration of the combination of tamsulosin-dutasteride combination the higher the PKC-α expression. Conclusion: Administration of long-term tamsulosin - dutasteride combination therapy causes upregulation PKC-α expression more than tamsulosin only.

Modulation of T-cell Regulators Associated with Advanced Stage of Prostate Cancer.

BACKGROUND: Benign prostatic hyperplasia (BPH) and prostate cancer are the most common prostate diseases. The possible role of the immune system in the pathogenesis of BPH and prostate cancer in recent years has begun to be widely studied. Although many studies have focused on T lymphocytes on the development of BPH and prostate cancer, the role of regulatory T-cells in the pathogenesis of BPH and prostate cancer is still not well known. OBJECTIVE: To determine the amount of regulatory T-cells in prostate cancer and BPH so that it can contribute to the concept of understanding the pathogenesis of prostate cancer and BPH. METHODS: This study used cross-sectional design study. Total samples were 24 patients, with 13 subjects prostate cancer group, and 11 subjects BPH group. Furthermore, peripheral blood samples are taken and then the amount of regulatory T-cells is calculated. After obtaining data on the amount of CD4+ CD25+ Foxp3+ regulatory T-cells in the blood, data analysis was performed between groups of patients diagnosed with prostate cancer and benign prostatic hyperplasia. RESULTS: The average amount of regulatory T-cells in the CRPC group was 53.44±29.43, prostate cancer group was 57.02±22.49 and the BPH group 89.71±9.31. One Way ANOVA test results showed that the average amount of regulatory T-cells between treatment groups gave a significant difference in regulatory T-cells with a p-value (0,003) <0.05. It can be concluded that there are differences in the average amount of regulatory T-cells, so we continued the testing with Tukey test. We continue to Pearson correlation study and resulted in significantly correlated with p value = 0.011 (P<0.05) and r = 0.414. CONCLUSIONS: It can be concluded there was significant difference between the average number of regulator T-cells in the BPH group compared with prostate cancer and CRPC patient. Further research is needed regarding the number of regulator T-cells CD4 + CD25 + FOXP3 + in prostate cancer patients (grouped according to Gleason score) and benign prostatic hyperplasia before and after therapy with bigger samples.

miRNA-21 Serum Evaluation in BPH, Hormone Sensitive Prostate Cancer, and Castrate Resistant Prostate Cancer: Attempt for Diagnostic Biomarker Evaluation.

BACKGROUND: Some of prostate cancer cases could progress to be Castrate Resistant Prostate Cancer (CRPC). However it is still a challenge to early diagnose it since no reliable examination could be done except PSA, which has high variability. It is now known that miRNAs are involved in nearly all inflammatory responses. Several malignancies in humans that specifically express miRNA have been detected and identified. The expression values of miRNA-21 also correlates with the occurrence of resistant castration of prostate cancer and metastases, therefore miRNA-21 is expected to be a biomarker to estimate the progression of cancer. OBJECTIVE: The purpose of this study was to analyze the expression values and cut-off markers of miRNA-21 as markers of CRPC progression. METHODS: This study used a retrospective cohort design with observational analysis. The forty-eight total sample was obtained from serum, then the RT-PCR was performed to obtain expression values of miRNA-21. Data were analyzed using One Way ANOVA to see the difference in the expression values of miRNA-21. Furthermore, to determine the cut-off analysis was carried out using the ROC curve. RESULTS: In the BPH group, an average expression value of miRNA-21 was 33,785±1.80 ng/dL, in the Prostate cancer group the average miRNA-21 was 34.51±1.32 ng/dL, while in the CRPC group, an average miRNA-21 was obtained, reaching to 34.51±1.32 ng/dL. The cut-off value of miRNA-21 from the BPH category was <33,595, PPV = 50%, NPV = 80% with a value of p = 0.081, the prostate Ca category was 33.595-35.21, PPV = 87.5%, NPV = 66.7% with p value = 0.003, while the value of miRNA-21 in the CRPC category was> 35.21, PPV = 80%, NPV = 58.3 with a value of p = 0.04. CONCLUSION: There is a significant difference in the expression values of miRNA-21 between BPH with CRPC and Prostate cancer and CRPC, therefore, miRNA-21 cut-off point is potential to differentiate the diagnosis.

miRNA-21 as Reliable Serum Diagnostic Biomarker Candidate for Metastatic Progressive Prostate Cancer: Meta-analysis Approach.

BACKGROUND: Prostate cancer is the second leading cause of cancer death in men, moreover when it develops metastasis. However, PSA detection in serum as current gold standard to measure disease progressivity had wide variability leading to confounding outcomes. MicroRNA-21 has diagnostic values for cancer over period of time researched, yet results are still inconclusive. OBJECTIVE: The aim of the study was to conduct recent meta-analysis to assess reliability of miRNA-21 as diagnostic biomarker especially in progressivity of prostate cancer. METHODS: Published papers from PubMed, Science Direct, and Embase" as of 1 July 2021 assessing circulating miRNA-21 in progressivity of prostate cancer patients were analyzed using Comprehensive Meta-Analysis tool. Pooled sensitivity, specificity, positive and negative likelihood ratio (LR) and SROC assessed with 95 % confidence intervals were estimated using fixed-effects or random-effects models. RESULTS: In total, we included 6 papers total of 651 samples reporting miRNA-21 capability of detecting progressive prostate cancer. The pooled sensitivity and specificity showed 0.91 (95% CI 0.88-0.94, I2=0%) and 0.89 (95% CI 0.85-0.92, I2=44.8%), respectively. Positive and negative likelihood ratio showed 7.18 (95% CI 4.31-11.96, I2=56%) and 0.11 (95% CI 0.07-0.16, I2=11.8%). SROC were assessed and got Area Under Curve around 97.4%. CONCLUSION: miRNA-21 could serve as biomarkers of prostate cancer progressivity since remarkable diagnostic value of circulating miRNA-21 in prostate cancer metastasis process.

In Silico and In Vitro Study: COX-2 Inhibition by Ethanol Extract of Dayak Onion Bulb (Eleutherine Americana Merr) as Treatment Innovation of Benign Prostatic Hyperplasia (BPH).

INTRODUCTION: Benign Prostatic Hyperplasia (BPH) is a benign tumor in males, which is histopathologically known with an increase of epithelial cells and prostatic stroma. Androgens, estrogens, stroma-epithelial interactions, growth factors, and chronic inflammation play a key role in the occurrence of BPH. Chronic inflammation in BPH is characterized by excessive expression of COX-2, which will trigger the expression of Bcl-2 anti-apoptotic protein. Dayak onion (Eleutherine Americana Merr) is a typical Kalimantan plant that is known as the treatment for prostate disease. This plant contains flavonoids which can inhibit the COX-2 enzyme thus causing a reduction in the production of prostaglandin E2. METHODS: This research was experimental research computationally and in vitro laboratory experimental research to determine COX-2 inhibitory activity by ethanol extracts of Dayak onion. RESULTS AND DISCUSSION: In in silico flavonoid, it was strongly related to COX-2 receptor on the active side of TYR371. Thus, it had the potential to inhibit COX-2. COX-2 inhibitor would cause BCL-2 to be inactive so that apoptosis occurr in BPH. In the in vitro research using human whole blood assay, the Dayak Onion bulb ethanol extract had IC50 COX-2 of 40.57 ng/ml and IC50 COX-1 of 364.89 ng/ml. Therefore, the ratio of IC50 COX-2 to IC50 COX-1 was 0.11. CONCLUSION: Ethanol extract of Dayak onion bulb has an inhibitory activity against COX-2. Thus, it has a potential of being an innovation for BPH treatment. Patient Summary: A healthy male, age 25-35 years old (history taking, physical and laboratory examination), and not using NSAIDs for the past 2 weeks.

Identification of afzelin potential targets in inhibiting triple-negative breast cancer cell migration using reverse docking.

BACKGROUND: Triple-negative breast cancer (TNBC) tends to be aggressive and metastatic, characteristics attributable to its cellular migration capabilities. Afzelin is a chemical compound with anti-metastatic potentials. This study aimed to predict proteins involved in TNBC cell migration which could be inhibited by afzelin. METHODS: The protein database was constructed from the Kyoto Encyclopedia of Genes and Genomes pathways collection which related to cell motility, then screened for druggability using SuperTarget and Therapeutic Target Database. The involvement of druggable proteins in the TNBC metastasis process was investigated through existing publications in The National Center for Biotechnology Information PubMed database. Inhibitory potential of afzelin toward target proteins was compared to the proteins' known-inhibitor, using the reverse docking method. RESULTS: Ten proteins identified as potential targets of afzelin, with the top 3 being ERK2, KRas, and FAK, respectively. Afzelin's 3-O-rhamnoside group played a dominant role in forming hydrogen bonds with the target proteins. Further analysis with STRING suggested that afzelin might be able to inhibit chemotaxis and haptotaxis of TNBC cells. CONCLUSIONS: Afzelin was predicted to inhibit TNBC cell motility, by targeting ERK2, KRas, and FAK activation.