Immunotherapy Applications for Thymine Dimers and WT1 Antigen in Renal Cancers: A Comparative Statistical Analysis.

Renal cell carcinoma (RCC) remains incurable in advanced stages. Biomarkers have proven to be quite useful in cancer therapeutics. Herein, we provide a comparative/integrative statistical analysis of seminal immunohistochemistry (IHC) findings for Wilms' Tumor 1 antigen (WT1) and thymine dimers (TDs), emerging as atypical, yet promising, potential biomarkers for RCCs. We assessed WT1/TD reactivity in adult RCC tumor cells, tumor microenvironment (TME), and tumor-adjacent healthy renal tissue (HRT). WT1 positivity was scarce and strictly nuclear in tumor cells, whereas TD-reactive tumor tissues were prevalent. We report statistically significant positive correlations between the density of reactive RCC cellularity and the intensity of nuclear staining for both biomarkers (WT1 - rho = 0.341, p-value = 0.036; TDs - rho = 0.379, p-value = 0.002). RCC stromal TME TD-positivity was much more frequent than WT1 reactivity, apparently proportional to that of the proper RCC cellularity and facilitated by extensive RCC inflammatory infiltration. TDs exhibited nuclear reactivity for most TME cell lines, while RCC TME WT1 expression was rare and inconsistent. In HRTs, TDs were entirely restricted to renal tubular cells, the likely cellular progenitor of most conventional RCC subtypes. In lieu of proper validation, these early findings have significant implications regarding the origins/biology of RCCs and may inform RCC therapeutics, both accounting for the high frequency of immunotherapy-permissive frameshift indels in RCCs, but also hinting at novel predictive clinical tools for WT1-targeted immunotherapy. Overall, the current study represents a meek yet hopefully significant step towards understanding the molecular biology and potential therapeutic targets of RCCs.

Risk Factors for Pulmonary Embolism in Individuals Infected with SARS-CoV2-A Single-Centre Retrospective Study.

The emergence of SARS-CoV2 has presented itself as a significant global health crisis. The prevalence of thrombotic events is known to be high in these patients, affecting various organ systems, sometimes leading to cutaneous thrombosis, pulmonary embolism (PE), stroke, or coronary thrombosis. The available evidence suggests that thromboembolism, hypercoagulability, and the excessive production of proinflammatory cytokines play a significant role in the development of multiorgan failure. Methodology: This retrospective single-centre study was conducted at "Victor Babes" University of Medicine and Pharmacy from Timisoara, Romania, involving a total of 420 patients diagnosed with COVID-19. We separated them into a CONTROL group that included 319 patients, and an intervention group (PE) with 101 patients that, subsequent to infection with the virus, developed pulmonary embolism. The study included the reporting of demographic data, laboratory findings, and comorbidities. Results: Out of a total of 420 patients, 24% experienced pulmonary embolism, while 21.42% died. Arterial thrombotic events were found to be associated with factors such as age, cardiovascular disease, levels of white blood cells, D-dimers, and albumin in the blood. The findings of the study indicate that there is an independent association between pulmonary thrombosis and hypertension (odds ratio (OR): 1.1; 95% confidence interval (CI): 0.7 to 1.7; p = 0.6463), cancer (OR: 1.1; 95% CI: 0.6 to 2.3; p = 0.6014), and COPD (OR: 1.2; 95% CI: 0.6 to 2.3; p = 0.4927). On the other hand, there is a stronger correlation between PE and obesity (OR: 2.8; 95% CI: 1.7 to 4.6; p < 0.0001), diabetes (OR: 3.3; 95% CI: 2 to 5.3; p < 0.0001), and dyslipidemia (OR: 3.6; 95% CI: 2.3 to 5.8; p < 0.0001) in a multivariable regression logistic model. Conclusions: Patients diagnosed with severe forms of COVID-19 display a comparable incidence of arterial thrombotic events, which have been linked to poor survival rates.

Colorectal Cancer Risk Prediction Using the rs4939827 Polymorphism of the SMAD7 Gene in the Romanian Population.

Colorectal cancer (CRC) is globally recognized as a prevalent malignancy known for its significant mortality rate. Recent years have witnessed a rising incidence trend in colorectal cancer, emphasizing the necessity for early diagnosis. Our study focused on examining the impact of the SMAD7 gene variant rs4939827 on the risk of colorectal cancer occurrence. The composition of our study group included 340 individuals, comprising 170 CRC diagnosed patients and 170 healthy controls. We performed genotyping of all biological samples using the TaqMan assay on the ABI 7500 Real-Time PCR System (Applied Biosystems, Waltham, MA, USA). This investigation focused on the rs4939827 gene variant, assessing its association with CRC risk and clinicopathological characteristics. Genotyping results for the SMAD7 gene variant rs4939827 revealed a 42.6% prevalence of the C allele in CRC patients (p = 0.245) and a 22.8% prevalence of the T allele in control subjects (p = 0.109). This study concluded that there was an elevated risk of CRC in the dominant model for CC/CT+TT, with a p-value of 0.113 and an odds ratio (OR) of 2.781, within a 95% confidence interval (CI) of 0.998 to 3.456. The findings of our research indicate a correlation between variants of the SMAD7 gene and the likelihood of developing colorectal cancer in our study population. Consequently, these results could be instrumental in facilitating early diagnosis of colorectal cancer utilizing information on single-nucleotide polymorphism (SNP) and genetic susceptibility to the disease.

Molecular Monitoring of Allogeneic Stem Cell Transplantation in Fanconi Anemia.

BACKGROUND: Allogeneic hematopoietic stem cell transplantation (HSCT) is the treatment of choice in patients with Fanconi anemia (FA). The aim of our study is to evaluate the impact and benefits of allogenic matched donor HSCT in a case of a 12 year-old girl with FA, who displayed good clinical evolution following 2 months post-transplantation. METHODS: In the pre-transplant phase, reference blood samples from the donor and recipient were collected on EDTA. The DNA from blood samples was extracted using an automated Maxwell® 48 RSC instrument (Promega, USA) with the Maxwell® RSC Whole blood DNA kit (Promega, USA). For DNA quantification, the PowerQuant System kit (Promega, USA) was used with the ABI 7500 Real-time PCR system (Applied Biosystems, USA). The amplification of the short tandem repeat markers was performed using the 24plex Investigator QS kit (Qiagen, Germany) on a ProFlex PCR System. Furthermore, the PCR products were separated and detected on an ABI 3500 Genetic Analyzer (Applied Biosytems, USA). RESULTS: Thirty days post transplantation, a complete chimerism (CC) was achieved with a full replacement by do-nor derived hematopoietic cells. Sixty days post transplantation, the CC status was maintained with improvement of hematological findings. CONCLUSIONS: In FA, chimerism monitoring after HSCT provides useful information regarding engraftment or possibility of post-transplantation complications such as graft versus host disease.

Chimerism Monitoring by Short Tandem Repeat (STR) Markers in Allogeneic Stem Cell Transplantation.

BACKGROUND: Allogeneic hematopoietic stem cell (allo-HSC) transplantation is used in the treatment of malignant hematological diseases. An important tool in monitoring post-transplantation evolution is represented by the percentage of donor's blood cells found in recipient's blood, known as chimersim. This is useful in predicting the graft rejection and the risk of disease relapse. In this study, we present the importance of multiplex STR markers in chimerism monitoring of a 8 year old girl diagnosed with acute lymphoblastic leukemia (ALL). METHODS: In the pre-transplant stage, saliva on buccal swabs and blood samples in EDTA were collected from the donor and recipient and used as reference samples. The DNA extraction from saliva and blood samples was done using the Pure Link Genomic DNA kit (Invitrogen, USA). For the DNA quantification, the Quantifiler Human DNA kit (Applied Biosystems, USA) was used on an ABI 7500 Real-time PCR system (Applied Biosystems, USA). Amplification of the STR markers was performed using the AmpFLSTR NGM SElect kit (Applied Biosystems, USA) on a ProFlex PCR System. The PCR products were separated and detected on an ABI 3500 Genetic Analyzer (Applied Biosytems, USA). RESULTS: One month post-transplantation of HSC, a mixed chimerism (MC) containing 38% of donor's cells was obtained from a bone marrow aspiration sample. On the 45th day, a new transplantation was performed. On the 15th day after 2nd transplantation, a MC with 91% donor's cells was obtained. On the 21st day after the 2nd transplantation, a complete chimerism (CC) with 100% donor's cells was obtained. CONCLUSIONS: Chimerism monitoring is useful in identifying those patients in risk for relapse or graft rejection.

Early Diagnosis of Oral Squamous Cell Carcinoma by Salivary microRNAs.

BACKGROUND: Oral squamous cell carcinoma (OSCC), the most common type of oral cancer, and represents more than 90% of malignancies of the oral cavity. Worldwide, each year about 275,000 are newly diagnosed. If detected at an early stage, OSCC has a survival rate of up to 80% compared to the detection in later stages (T3-T4) when a survival rate of 20 - 30% is present. METHODS: Because OSCC presents these survival rates, there is an urgent need to introduce new non-invasive molecular biomarkers for the early detection of OSCC from saliva, which will contribute to an increased long term survival rate for these patients. RESULTS: MicroRNAs represent small, non-coding RNAs that have important roles in biochemical mechanisms, carcinogenesis, cell proliferation, embryogenesis, and other mechanisms involved at the molecular level in the functioning of the human body. CONCLUSIONS: In the last decade, due to the fact that forensic genetics developed significantly, salivary microRNAs were increasingly studied as non-invasive molecular biomarkers which could aid in early diagnosis, monitoring, and prognosis of oral cancers. This review will present the most important salivary microRNAs which are involved in oral carcinogenesis, especially those which could be used as potential biomarkers in early detection, monitoring, and prognosis of oral cancers by non-invasive techniques.

Molecular Diagnostic of Prostate Cancer From Body Fluids Using Methylation-Specific PCR (MS-PCR) Method.

BACKGROUND: Worldwide prostate cancer (PCa) represents the 2nd leading cause of cancer related deaths among men. Currently, the screening for early detection of PCa is based on determination of serum prostate-specific antigen (PSA) levels. But this biomarker presents some disadvantages related to its specificity and sensitivity. In our study, we want to determine if methylation levels of the glutathione S-transferase P1 (GSTP1) gene could be used as a new biomarker for the early detection of PCa and to distinguish between malignant and benign pros-tatic lesions. METHODS: To determine the methylation levels of the GSTP1 gene, 31 men with histopathological diagnosis of prostate adenocarcinoma and 34 men with the histopathological diagnosis of benign prostatic hyperplasia (BPH) as controls were included in the study group. The genomic DNA was extracted from urine samples. We analyzed the methylation levels of the GSTP1 gene by methylation-specific polymerase chain reaction (MS-PCR) method. RESULTS: In prostate cancer patients 27 of 31 (87%) presented hypermethylated levels of the GSTP1 gene, whereas 4 of 34 (11.8%) BPH patients had hypermethylated levels of the GSTP1 gene. Further, in the case of these four patients a second biopsy was done, which confirmed the diagnosis of prostate adenocarcinoma. Using the receiver operating curve (ROC), we obtained a specificity of 87% and a sensitivity of 98% for the GSTP1 gene. CONCLUSIONS: We can conclude that GSTP1 represents a new molecular biomarker which can aid in early detection of PCa and be used to discriminate between benign and malignant prostatic lesions from body fluids by noninvasive methods.

Modulation of the Redox Expression and Inflammation Response in the Critically Ill Polytrauma Patient with Thoracic Injury. Statistical Correlations between Antioxidant Therapy.

BACKGROUND: One of the major causes of mortality in the world is represented by multiple traumas. Thoracic trauma is commonly associated with polytraumas. A series of physiopathological complications follow polytraumas, leading to a significant decrease in the survival rate. As a result of injuries, significant quantities of free radicals (FR) are produced, responsible for oxidative stress (OS). To minimize the effects of OS, we recommend the administration of antioxidant substances. In this study we want to highlight statistically significant correlations between antioxidant therapy and a series of clinical variables. METHODS: This retrospective study included 132 polytrauma patients admitted to the ICU-CA between January 2013 and December 2014. The selection criteria were: injury severity score (ISS) ≥ 16, ≥ 18 years, presence of thoracic trauma (abbreviated injury scale, AIS ≥ 3). Eligible patients (n = 82) were divided into two groups: Group 1 (n = 32, antioxidant free, patients from 2013) and Group 2 (n = 50 antioxidant therapy, patients from 2014). Antioxidant therapy consisted in the administration of vitamin C (i.v.), vitamin B1 (i.v.), and N-acetylcysteine (i.v.). Clinical and biological tests were repeated until discharge from ICU-CA or death. RESULTS: Between Group 1 and Group 2 statistically significant differences were highlighted regarding the ISS score (p = 0.0030). 66% of patients from Group 2 were admitted at more than 24 hours after the trauma, in contrast to the patients from Group 1, where 62.5% were directly admitted to the ICU (p = 0.0114). Compared with the patients from Group 1, patients who received antioxidant therapy show improved parameters: leukocytes (p < 0.0001), platelets (p = 0.0489), urea (p = 0.0199), total bilirubin (p = 0.0111), alanine transaminase (p = 0.0010), lactat dehydrogenase (p < 0.0001). Between the two groups there were no statistically significant differences regarding the length of stay in the ICU-CA (p = 0.4697) and mortality (p = 0.1865). CONCLUSIONS: Following the study, we can affirm that due to the administration of antioxidant substances, posttraumatic complications are greatly reduced. Moreover, the administration of high dose of antioxidants remarkably improves the clinical status of the critical patient.

Renal Cancer Diagnosed by Noninvasive Methods from Body Fluids by Quantitative Methylation-Specific PCR(qMSP).

BACKGROUND: Renal cell carcinoma (RCC) represents the 9th most common malignancy in the world, having an incidence peak in the range of 60 to 70 years of age. Most of these malignancies are detected in an advanced stage. Thus, there is an urgent need for developing new tools composed of biomarkers. METHODS: In the present study we measured the promoter methylation of the Ras association domain family 1A gene (RASSF1A) by quantitative methylation-specific PCR (qMSP) in paired urine samples from 13 RC patients and from 13 corresponding controls. RESULTS: In RC patients, only 2 of 13 (15.4%) were unmethylated, whereas 11 of 13 (84.6%) were methylated. In the control group all the subjects were unmethylated. We analyzed the receiver operating curve (ROC) and obtained a sensitivity of 84.6% and a specificity of 100%, respectively, for the RASSF1A gene. The area under the curve (AUC) was of 0.923. CONCLUSIONS: Being involved in the initiation and progression of renal carcinogenesis, RASSF1A gene could aid as a biomarker in the early detection of renal cancer, its prognosis, and in its follow-up.

Genetic and Epigenetic Biomarkers of Molecular Alterations in Oral Carcinogenesis.

Worldwide, oral cancers represent the 6th most common type of cancer. Oral squamous cell carcinoma (OSCC), which is the most common type of oral cancer, is present in about 90% of the patients with this malignancy. OSCC presents a survival rate up to 80%, if it is detected in an early stage (T1), but if detected at later stages (T3 - T4) the survival rate decreases to 20 - 30%. Due to these survival rates, it is obvious that there is an urgent need to introduce new molecular biomarkers for the early, noninvasive diagnosis of oral cancers from saliva. These biomarkers will aid in increasing the survival rate of the patients for the long-term. MicroRNAs are part of a class of small, non-coding RNAs that contain 19 - 23 nucleotides. MicroRNAs play an important role in the regulation of biochemical mechanisms, cell proliferation, and other cellular mechanisms in the human body. Recently, due to the developments in the field of molecular genetics, salivary microRNAs became important biomarkers in early detection and monitoring of oral cancers by noninvasive methods. We want to present in this review the most important genetic and epigenetic biomarkers involved in oral carcinogenesis, focusing especially on the salivary microRNAs as biomarkers in early diagnosis of OSCC.

Prostate cancer molecular detection in plasma samples by glutathione S-transferase P1 (GSTP1) methylation analysis.

BACKGROUND: Prostate cancer (PCa) represents the most commonly diagnosed type of malignancy among men in Western European countries and the second cause of cancer-related deaths among men worldwide. Methylation of the CpG island has an important role in prostate carcinogenesis and progression. The purpose of the study was to analyse the diagnostic value of aberrant promoter hypermethylation of the gene for glutathione S-transferase P1 (GSTP1) in plasma DNA to discriminate between prostate cancer (PCa) and benign prostatic hyperplasia (BPH) patients by minimally invasive methods. METHODS: Aberrant promoter hypermethylation was investigated in DNA isolated from plasma samples of 31 patients with diagnostic of PCa and 44 cancer-free males (control subjects). Extracted genomic DNA was bisulfite treated and analyzed using methylation-specific polymerase chain reaction (MS-PCR) technique. RESULTS: Hypermethylation of the GSTP1 gene was detected in plasma samples from 27 of 31 (92.86%) patients with PCa. Genomic DNA from plasma samples from the 44 controls without genitourinary cancer revealed promoter hypermethylation of GSTP1 gene in 3 (10.6%) of the 44 patients. Receiver operating curve (ROC) included clinico-pathological parameters such as: serum PSA levels, pathological stage, Gleason score, hypermethylation status of GSTP1 gene, and it gave a predictive accuracy of 93% with a sensitivity and specificity of 95% and 87%, respectively. CONCLUSIONS: In this study, we have evaluated the ability of GSTP1 gene to discriminate between PCa and BPH patients in genomic DNA from plasma samples by non-invasive methods.

The role of prostatic inflammation biomarkers in the diagnosis of prostate diseases.

Benign prostatic hyperplasia (BPH) and prostate cancer (PCa) are chronic conditions, which are hormone-dependent and epidemiologically associated with prostate inflammation. As a large number of studies have demonstrated, the stimulation of T-cells at the level of prostatic chronic inflammatory infiltrates is followed by stromal and epithelial cell proliferation. The aim of this review is to present the actual level of knowledge in the field of prostatic immune response and chronic inflammation, and to analyze the relationships between chronic inflammation and BPH/PCa. The most studied prostatic inflammation biomarkers detected in biological fluids are also presented, together with their potential roles in the diagnosis and prognosis of prostatic disease.

Genetic and epigenetic biomarkers for early detection, therapeutic effectiveness and relapse monitoring in bladder cancer.

Urinary bladder cancer is the fifth most common cancer in the Western world and is responsible for about 3% of all cancer-related deaths. Because most advanced invasive or metastatic cancers have low cure rates, risk assessment and early detection of the clinically occult premalignant phases of neoplasia are a particular importance. Many tumor biomarkers for bladder cancer have been evaluated for use in detecting and monitoring bladder cancers tissue specimens, bladder washes, and urine specimens but, none of the biomarkers reported to date has shown sufficient sensitivity and specificity to detect the entire spectrum of bladder cancers in routine clinical practice. The limitations of established prognostic markers requires us to identify better molecular parameters that could be of interest in predicting the prognosis of bladder cancer patients, in particular, the high-risk patient groups that are at risk of progression and recurrence. Methylation is an important molecular mechanism in the development of bladder cancer and could be used as a prognostic and diagnostic biomarker, because hypermethylation of several gene promoters was detected in urine sediment DNA from bladder cancer patients. Aberrant patterns of epigenetic modification could be, in the near future, crucial indicators in cancer diagnosis, prognosis, and additionally could be good targets for developing novel therapies while maintaining quality of life.

Clinical applications of molecular biomarkers in prostate cancer detection.

Prostate cancer is a heterogeneous disease with regard to molecular alterations and clinical course. Early diagnosis of prostate cancer can increase the curative success rate for this disease. Because of the recent developments in the field of molecular biology, an increased interest occurred for molecular biomarkers, as tools for early prostate cancer detection, monitoring disease progression, predicting disease recurrence and therapeutic treatment efficacy. Many molecular biomarkers have been discovered in human serum, urine, seminal fluid and histological specimens.

Applicabillity of noninvasive biomarkers in prostate cancer diagnosis.

Prostate cancer represents the second leading cause of male cancer-related deaths worldwide. Better indicators of the presence of prostate cancer are needed to avoid unnecessary treatment, predict disease course and develop more effective therapy. Many molecular biomarkers have been described in human serum, urine, seminal fluid and histological specimens that exhibit varying capacities to detect prostate cancer and predict disease course.

The importance of clinical application of molecular biomarkers in bladder cancer detection.

It is known that high throughput technologies facilitate the identification of new molecular targets and biomarkers specific for bladder cancer.The new field of molecular medicine promises that clinical outcomes will be improved by directing therapy toward the molecular mechanisms and targets associated with the growth of the patient's tumor.The great challenge remains to improve the measurement of these targets and to translate this wealth of discovery into clinical management of bladder cancer.