Extreme Activation of Androgen Receptor for Prostate Cancer Therapy

Prostate cancer (PCa) is the second most common cancer worldwide in men and one of the major causes of cancer-related death among men in Australia. In PCa cells, the androgen receptor (AR) is the key driver of cell proliferation, cell cycle progression, and metabolism; thus, blocking AR activity with androgen deprivation therapy (ADT) is a standard-of-care treatment for metastatic PCa. However, ADT is never curative, with all patients eventually relapsing with lethal castration-resistant prostate cancer (CRPC). In a paradoxical phenomenon, potent activation of AR with high doses of androgens can also inhibit the growth of PCa tumours. However, the exact mechanism(s) by which activation of AR can block PCa growth is poorly understood. Therefore, in my PhD project, I explored the mechanisms underlying PCa growth suppression in response to extreme activation of AR using a potent androgen, methyltestosterone (MeT). I have found that methyl-testosterone (MeT), a synthetic androgen, can potently transactivate AR and suppress the proliferation of AR-positive prostate cancer cells (LNCaP, C42B, MR49F, and 22RV1) but not an AR-negative cell line (PC3) or a PCa model expressing a version of the AR lacking the ligand-binding domain (R1-D567), suggesting that the growth-inhibitory effects of MeT are AR-dependent. Mechanistically, MeT acts much like high-dose dihydrotestosterone (DHT) in terms of genome-wide AR binding (evaluated by ChIP-seq) and the transcriptional program activated via AR (evaluated by RNA-seq). However, these analyses showed that MeT only extends the AR cistrome and enables AR to act as a potent transcriptional repressor of genes associated with cell cycle, DNA replication, and DNA damage responses. Unexpectedly, our RNA-seq data revealed that MeT dysregulates the expression of transposable elements, including endogenous retroviruses (ERVs). Mechanistically, we found that MeT suppresses the expression of DNA methyl-transferases (DNMTs) and EZH2, which are considered to be key factors repressing the expression of transposable elements. Consistent with the proposed hypothesis, my PhD work showed that MeT caused global hypomethylation of DNA and re-distribution of H3K27me3. More specifically, my research supports a model whereby DNA hypomethylation was linked to the induction of endogenous retroviruses (ERVs). Interestingly, I found that ERV induction was associated with a “viral mimicry” response characterised by activation of pattern recognition receptors RIG-I and STING and subsequent activation of interferon (IFN) signalling. Importantly, I also observed increased expression of MHC class I genes with MeT treatment, suggesting that it can enhance tumour immunogenicity. Validating this finding, co-culture of a murine model of PCa (RM1) with tumour-specific CD8+ T cells revealed that MeT promoted enhanced recognition and functional cytokine production by T cells. Collectively, my work has provided a greater understanding of growth-inhibitory effects of androgens on PCa tumours and uncovers a potential new role for high-dose androgen therapy as an immunosensitisation agent.Thesis (Ph.D.) -- University of Adelaide, Adelaide Medical School, 202

Coronary Artery Bypass Graft in Six Members of a Family: A Case Series

Abstract: Coronary artery disease (CAD) is a multifactorial problem. Although hyperlipidemia (HLP), diabetes (DM), and hypertension (HTN) are known as the familial and cardiac risk factors, CABG is rare in the several members of a family. We reported the six members of a family who presented with MI and/or advanced angina pectoris during the 6 years. CABG(Coronary artery bypass graft) considered for all patients due to involvement in main coronary arteries and clinical presentation. The ejection fraction (EF) was varied between 25% to 70%. Beating heart cardiopulmonary bypass (beating heart CPB) and off-pump were chosen as surgical method. The mean duration of surgery was about 4 hours. The only post-operative complication was deep wound infected in a young man who was smoker and drug abuser. According to family relationship of patients and the high effect of genetic penetration, we presume the gene influence to incidence CAD and potential to undergoing CABG, in members of a special family

Supportive care in a patient with Alstrom syndrome with hyperphenylalaninemia and sleep problems

Abstract Alstrom syndrome is a rare genetic disorder with an autosomal recessive mutation in the ALMS1 gene. The disease's manifestations include ophthalmic problems, hearing loss, obesity, and cardiovascular disorders. In addition, medical cases include other organ complications. However, the overlapping variety of such symptoms with other diseases may delay the diagnosis. In this article, we describe the case of a 7‐year‐old female patient with Alstrom syndrome, and cardiovascular and hyperphenylalaninemia diseases since birth. Other symptoms included diabetes and ophthalmologic problems with skeletal disability. Blindness and hearing impairment were diagnosed, along with recurrence of respiratory problems at the age of 7 years. The patient's obesity‐induced snoring predisposed her to uncontrolled blood glucose. In fact, respiratory tract problems and sleep disorders had occurred as a degraded cycle and left her with a severe disability for years. The similarity of the symptoms with other diseases had misled the physician in diagnosis. However, a polysomnography test (because of complaints of short sleep duration) recognized the source of the patient's sleep disorders and breathing problems. Eventually, we delivered a portable ventilator to the child for continuous positive airway pressure (CPAP) therapy. The child's breathing and oxygenation conditions improved. Using the ventilator and the CPAP system, we discharged her from the hospital without requiring oxygenation, in a stable condition. The procedure could prevent the patient from hypoxia and retinal problem

Evaluation of the physicochemical and microbial properties of lamb meat coated with Shirazi balangu seed mucilage-based edible coating containing cell-free supernatant of Levilactobacillus brevis G145

This study was aimed to design a novel edible coating based on Shirazi balangu seed mucilage (SBM) containing cell-free supernatant (CFS) of Levilactobacillus brevis G145 to improve the quality and shelf-life of lamb meat slices during cold storage. The CFS (1 and 2 %) was added to the SBM, and the obtained edible coating was used to coat the meat slices. The samples were stored at 4 °C for 1, 4, 7, and 10 days. In comparison to the control sample, the meat slices coated with SBM+2 %CFS had manifestly lower total viable count (6.62 vs. 13.10 log CFU/g), psychrotrophic count (3.47 vs. 8.80 log CFU/g), Escherichia coli count (0.80 vs. 1.54 log CFU/g), Staphylococcus aureus count (0.81 vs. 1.57 log CFU/g), fungi count (0.68 vs. 2.38 log CFU/g), pH (5.70 vs. 6.51), and peroxide value (4.89 vs. 10.80 meq O2/kg), and higher moisture content (70 vs. 60 %), hardness (67.80 vs. 57.60 N), and sensory properties by the end of storage period. The results of this study revealed that the SBM+CFS- based edible coating increased the shelf-life of lamb meat efficiently

Recent advances on biomedical applications of scaffolds in wound healing and dermal tissue engineering.

The tissue engineering field has developed in response to the shortcomings related to the replacement of the tissues lost to disease or trauma: donor tissue rejection, chronic inflammation and donor tissue shortages. The driving force behind the tissue engineering is to avoid the mentioned issues by creating the biological substitutes capable of replacing the damaged tissue. This is done by combining the scaffolds, cells and signals in order to create the living, physiological, three-dimensional tissues. A wide variety of skin substitutes are used in the treatment of full-thickness injuries. Substitutes made from skin can harbour the latent viruses, and artificial skin grafts can heal with the extensive scarring, failing to regenerate structures such as glands, nerves and hair follicles. New and practical skin scaffold materials remain to be developed. The current article describes the important information about wound healing scaffolds. The scaffold types which were used in these fields were classified according to the accepted guideline of the biological medicine. Moreover, the present article gave the brief overview on the fundamentals of the tissue engineering, biodegradable polymer properties and their application in skin wound healing. Also, the present review discusses the type of the tissue engineered skin substitutes and modern wound dressings which promote the wound healing

Detection of Merkel cell polyomavirus large T-antigen sequences in human central nervous system tumors

Despite decades of epidemiological investigation, little is known about the etiology of the central nervous system (CNS) tumors, and few well-established risk factors have been recognized. This study tested the presence of Merkel cell polyomavirus (MCPyV), the only member of the Polyomaviridae family convincingly linked to human cancer, in diverse CNS malignancies. In total, 58 CNS tumor biopsies were analyzed for the MCPyV large T-antigen (LT-Ag) gene by quantitative real-time PCR. Merkel cell polyomavirus LT-Ag DNA load was determined as viral copies per cell and viral copies per microliter of purified genomic DNA from CNS tumor samples. The MCPyV LT-Ag sequence was detected in 34 (58.6) of the 58 tested samples. Viral LT-Ag was quantified in 19.0 of schwannomas, 13.8 of meningiomas, and 5.2 of pituitary adenomas. The difference between MCPyV positivity in different types of CNS malignancies was not statistically significant (P=0.066). The mean LT-Ag copy number in 34 positive samples was 744.5±737.7 and 0.056�10-3±0.091�10-3 per microliter and per cell, respectively. Among MCPyV-positive CNS tumors, the mean MCPyV copy number was higher in meningiomas (993.8±853.2 copy per microliter and 0.098�10-3±0.108�10-3 copy per cell). Multiple linear regression analysis revealed statistically significant difference in MCPyV copy number between meningioma and other CNS tumor types, when the model was adjusted for age and sex (P=0.024). This study shows the first evidence of the detection of MCPyV LT-Ag sequence at a low copy number in human CNS tumors. © 2015 Wiley Periodicals, Inc

Prevalence of JC polyomavirus large T antigen sequences among Iranian patients with central nervous system tumors

The human neurotropic JC virus (JCV) is of significant interest due to its experimental neuro- oncogenic potential. In clinical samples from human central nervous system (CNS) tumors, detection of JCV sequences suggests a possible association with CNS neoplasms, but the results are discrepant worldwide. To assess the prevalence of JCV sequences in Iranian patients with primary and metastatic CNS malignancies, a total of 58 fresh CNS tumors were examined by quantitative real-time PCR targeting the JCV large T antigen (LT-Ag) gene, and JCV DNA load was determined as viral copy number per cell. All patients were immunocompetent, and none of them had received immunosuppressive therapy before surgical operation. JC virus LT-Ag sequences were found in a total of 15 (25.9 ) out of the 58 tested samples. In primary CNS tumors, JCV sequences were identified more frequently in meningiomas (50.0 ) and schwannomas (35.7 ). In metastatic CNS tumors, JCV LT-Ag was identified in one case with brain adenocarcinoma originating from lung cancer. No statistically significant association between JCV positivity and various types of CNS malignancies was observed (P = 0.565). The mean JCV LT-Ag copy number in 15 positive cases was 1.8 � 10�4 ± 4.5 � 10�4 copies per cell (range 1.0 � 10�5-1.78 � 10�3 copies per cell). An inverse correlation between white blood cell (WBC) count and JCV copy number was observed, but this correlation was not statistically significant (R = �0.198, P = 0.480). This study provides the first data on the prevalence of JCV in primary and metastatic CNS tumors from Iranian patients. © 2014, Springer-Verlag Wien

PCR-free paper-based nanobiosensing platform for visual detection of telomerase activity via gold enhancement

© 2020 Elsevier B.V. Telomerase activity has been demonstrated in a wide variety of most solid tumors and considered as a well-known cancer biomarker. The commonly utilized method for its detection is polymerase chain reaction (PCR)-based telomeric repeat amplification protocol (TRAP). However, the TRAP technique suffers from false-negative results caused by the failure of PCR step. Moreover, it requires advanced equipment with a tedious and time-consuming procedure. Herein, we presented a portable nitrocellulose paper-based nanobiosensing platform for ultrafast and equipment-free detection of telomerase activity based on a simple colorimetric assay that enabled naked-eye visualization of the color change in response to enzyme activity. In this platform, hybridization was initially performed between telomere complementary oligonucleotide immobilized on gold nanoparticles (GNPs) and telomerase elongated biotinylated probe. Thereafter, the assembly was attached on activated paper strip via avidin-biotin interaction. The signal amplification was carried out by enlargement of the attached GNPs on the paper strip, forming tightly compact rod-shaped submicron structures of gold representing a visual color formation. Thanks to significant sensitivity enhancement, the color change was occurred for down to 6 cells, which can be easily observed by the naked eye. Due to the desired aspects of the developed assay including PCR-free, low cost, simple, and high sensitivity, it can be used for evaluation of telomerase activity in cell extracts for future clinical applications. Furthermore, this design has the ability to be easily integrated into lab-on-chip devices for point-of-care telomerase sensing