Role of Melatonin in Angiotensin and Aging.

The cellular utilization of oxygen leads to the generation of free radicals in organisms. The accumulation of these free radicals contributes significantly to aging and several age-related diseases. Angiotensin II can contribute to DNA damage through oxidative stress by activating the NAD(P)H oxidase pathway, which in turn results in the production of reactive oxygen species. This radical oxygen-containing molecule has been linked to aging and several age-related disorders, including renal damage. Considering the role of angiotensin in aging, melatonin might relieve angiotensin-II-induced stress by enhancing the mitochondrial calcium uptake 1 pathway, which is crucial in preventing the mitochondrial calcium overload that may trigger increased production of reactive oxygen species and oxidative stress. This review highlights the role and importance of melatonin together with angiotensin in aging and age-related diseases.

Role of melatonin in the treatment of COVID-19; as an adjuvant through cluster differentiation 147 (CD147).

COVID-19 caused by the SARS-CoV-2 outbreak quickly has turned into a pandemic. However, no specific antiviral agent is yet available. In this communication, we aimed to evaluate the significance of CD147 protein and the potential protective effect of melatonin that is mediated by this protein in COVID-19. CD147 is a glycoprotein that is responsible for the cytokine storm in the lungs through the mediation of viral invasion. Melatonin use previously was shown to reduce cardiac damage by blocking the CD147 activity. Hence, melatonin, a safe drug, may prevent severe symptoms, reduce symptom severity and the adverse effects of the other antiviral drugs in COVID-19 patients. In conclusion, the use of melatonin, which is reduced in the elderly and immune-compromised patients, should be considered as an adjuvant through its CD147 suppressor and immunomodulatory effect.

Programming hMSCs into Potential Genetic Therapy in Cancer.

Based on their distinct characteristics, such as self-renewal and differentiation potential, human mesenchymal stem cells (hMSCs) have been proposed as a feasible tool for cancer therapy. The characteristic of hMSCs that can be used in cancer therapy is their ability to home to primary and metastatic tumor sites. Recent studies have shown that use of stem cells obtained from adult tissue may be a novel vehicle for stem cell-mediated cancer therapy with improved antitumor effects. Stem cells have been used as vehicles to deliver various agents to tumor sites in order to decrease the size of the tumor or increase the lifespan of the organism. Genetically modified MSCs have been shown to increase apoptosis and decrease growth and angiogenesis in solid tumors. In this review, we will focus on the potential of the genetically modified hMSC-based genetic therapy that is a combination of stem cell and gene therapy approaches and its potential advantages over current therapies.

Role of Mesenchymal Stem Cells in Cancer Development and Their Use in Cancer Therapy.

Stem cells have the ability to perpetuate themselves through self-renewal and generate mature cells of a particular tissue through differentiation. Mesenchymal stem cells (MSCs) play an important role in tissue homeostasis - supporting tissue regeneration. MSCs are rare pluripotent cells supporting hematopoietic and mesenchymal cell lineages. MSCs have a great therapeutic potential in cancer therapy, as well as stem cell exosome and/or microvesicle-mediated tissue regeneration. In this review, the use of hMSCs in stem cell-mediated cancer therapy is discussed.

PRACTICE OF CONSANGUINITY AND ATTITUDES TOWARDS RISK IN THE PASHTUN POPULATION OF KHYBER PAKHTUNKHWA, PAKISTAN.

This study aimed to investigate the frequency of consanguineous marriages and level of understanding of consanguinity-associated genetic risks in the Pashtun population, Pakistan. Information was gathered using a detailed questionnaire completed by 1500 individuals of both sexes over the 11-month period between April 2015 and February 2016. The mean inbreeding coefficient of the respondents was calculated and a five-point response scale was used to assess their understanding of consanguinity risks. The frequency of consanguineous marriages in the Pashtun population was found to be 58.3%, with a mean inbreeding coefficient of 0.0259. Marriage between second cousins was found to be the dominant marriage type. Level of education was found to be negatively related to the incidence of consanguineous marriage (p<0.001), and higher consanguinity was reported among the rural than the urban population (p<0.001). Participants in the ≥25-year age group, those with a higher level of education and those residing in urban areas exhibited a significantly higher understanding of consanguinity risks. The overall prevalence of consanguinity in the Pashtun population is high, demonstrating the need for awareness of its risks in the target population. The timely dissemination of information on potential health-related risks and the introduction of genetic counselling in the region would benefit both the individuals concerned and the community in general.

Modeling Mesenchymal Stem Cells in TMJ Rheumatoid Arthritis and Osteoarthritis Therapy.

Stem cells have self-renewal capacity and an ability to differentiate into particular cell types generating mature cells. Mesenchymal stem cells (MSCs) have a significant role in tissue homeostasis, which leads into tissue regeneration. MSCs are rare pluripotent cells supporting hematopoietic and mesenchymal cell lineages. MSCs are also believed to have therapeutic power over temporomandibular joint (TMJ) disorders (TMDs). The most common type of TMD is articular disc displacement, which induces progressive degenerative changes. These changes lead to rheumatoid arthritis or osteoarthritis. In this review, use of human mesenchymal cells (hMSCs) for therapeutic treatment of inflammatory diseases of TMJ is discussed.

Localization of the Werner Protein Together with H2AX in γ-Irradiation-Induced Neoplastic Transformed Human Mesenchymal Stem Cells.

The H2A histone family, member X (H2AX), and Werner (WRN) are important proteins for genome and telomere maintenance. WRN has a major role in genome stability, particularly during DNA replication, transcription, recombination, and repair of DNA double-stranded breaks (DSBs) via base excision repair, homologous recombination, or nonhomologous end joining. H2AX plays a part in the rapid, sensitive, cellular response to the ionizing radiation or DNA-damaging chemotherapeutic agents that cause DSBs. This occurs when radiation-induced DSBs trigger the activation of H2AX and begin the damage-repair process. In this study, we investigate the role and localization of WRN together with DNA damage marker H2AX at the radiation-induced damaged sides of both the telomere-immortalized human mesenchymal stem cells (hMSCs) and hMSC-telomere 1 (hMSC-telo1) and in control primary hMSCs. Phosphorylated H2AX and WRN immune staining enabled evaluation of overall genomic integrity and damage/repair. We used peptide nucleic acid-fluorescent in situ hybridization to visualize telomeric damage as a short-term effect. A high-level WRN signal was observed in both primary hMSCs and telomerase-immortalized hMSCs after the cells had been subjected to infrared radiation. Afterward, the irradiation level of the WRN signals decreased considerably, especially in later passages, and WRN was nondetectable in the latest passages of the hMSC Telo1 cells. Contrary to this finding, we found that levels of H2AX phosphorylation in hMSC-telo1 cells increased with time, especially at telomere sides, suggesting that cells with long telomeres and high telomerase activity have the advantage of maintaining genomic integrity. Evaluation of localization of WRN signals demonstrated that WRN does not leave the nucleolus after irradiation. We did not detect the WRN signal at the telomere sides, but we could detect H2AX at the telomeric sides. Thus, our overall data suggest that the WRN protein is not involved in irradiationinduced DNA damage/repair, even at telomeric sides in hMSC and hMSC-telo1.

The Route to HPV-Associated Neoplastic Transformation: A Review of the Literature.

Human papillomaviruses (HPVs)-small, nonenveloped viruses with double-stranded circular DNA-are believed to have a role in the progression of cancer. However, the exact mechanisms are not well established. The interference of HPV proteins, especially E6 and E7, in the cell cycle is considered to be the main pathway. It is still questioned whether the expression of these proteins or the viral load is more important in neoplastic transformation. Furthermore, HPV is believed to adapt mechanisms to evade the host cell immune system; persistent HPV infection may also play a role in oncogenic transformation by causing genomic instability and local immune suppression. These factors may cause accumulation of genomic alterations within the host cell and integration of the viral genome into the host genome. In recent years, epigenetic modifications, such as methylation, have also been considered to take part in neoplastic transformation. All of these alterations to the genome may be favorable to the development of cancer. This article highlights the association of HPV in neoplastic transformation and cancer progression.

Human Mesenchymal Stem Cells in Cancer Therapy.

Human mesenchymal stem cells (hMSCs) have the ability to differentiate into several tissue types. Their use in cancer therapeutics or as therapeutic delivery vehicles has significant potential, particularly in their exosome/microvesicle-mediated tissue regeneration abilities. In this review, the potential use of hMSCs in cancer therapy is discussed.

Expression of TRF2 and its prognostic relevance in advanced stage cervical cancer patients.

BACKGROUND: Telomeres are protective caps consisted of specific tandem repeats (5'-TTAGGG-3'). Shortening of telomeres at each cell division is known as "mitotic clock" of the cells, which renders telomeres as important regulators of lifespan. TRF2 is one of the critical members of shelterin complex, which is a protein complex responsible from the preservation of cap structure, and loss or mutation of TRF2 results in DNA damage, senescence or apoptosis. Since cancer is frequently associated with aberrant cell cycle progression, defective DNA repair or apoptosis pathways, TRF2 could be one likely candidate for cancer therapy. Here we investigated the prognostic role of TRF2 levels in cervical cancer patients. Fold-induction rates were evaluated with respect to median values after real-time PCR analysis. Overall survival, distant disease-free and local recurrence-free survival rates were calculated using Kaplan-Meier long rank test. RESULTS: Both five year overall- and disease-free survival rates were longer in patients with higher TRF2 expression compared to lower expression, but results were not statistically significant (69.2% vs 28.9%, respectively). Mean local recurrence-free survivals (LRF) were very close ( 58.6, CI: 44.3-72.9 vs 54.5, CI: 32.1-76.9 months) for high and low expressions, respectively. Cumulative proportion of LRF at the end of five year period was 76.9% for high and 57.1% for low TRF2 expression (P = 0.75). Statistically significant difference was found between survival ratios and Bcl-xL and p53 gene expressions, but not with TRF2. A respectable correlation between TRF2 expression and apoptosis along with distant metastasis was noted (P = 0.045 and 0.036, respectively). Additionally, high TRF2 expression levels had a positive impact in five year survival rate of stage IIIB-IVA patients (P = 0.04). CONCLUSIONS: Our results support the role of TRF2 in apoptosis and imply a positive relation with distant metastases and survival in advanced stage patients. The remarkable difference in survival periods of patients with different TRF2 expressions suggest that TRF2 may be a candidate factor to estimate survival for cervical cancer, a preliminary observation which should further be verified with a larger cohort.

Performance of risk prediction models in breast cancer screening among women in Cyprus.

Background: Tools that can predict breast cancer risk in women will have a significant impact on women and healthcare systems in low- and middle-income countries. Aims: We compared the performances of the Breast and Ovarian Analysis of Disease Incidence and Carrier Estimation Algorithm (BOADICEA) model, the International Breast Cancer Intervention Study (IBIS) model, and the Gail model in predicting breast cancer risk among women in Cyprus. Methods: We recruited 655 women from Dr Burhan Nalbantoglu Devlet Hastanesi Hospital in Lefkosa: 318 had breast cancer and 337 did not have breast cancer (hospital-based controls). We collected retrospective data from the hospital's medical records and interviews with the women after informed consent. Data collected included: age, age at diagnosis, age at menarche, menopausal status, presence of benign breast disease, breast cancer in relatives, BRCA-1 and BRCA-2 mutations, history of hormone replacement therapy, and breast density. We calculated the 5-year risk of breast cancer and the risk values were used to plot receiver operating curves. Results: For the area under the curve (95% confidence interval, CI), sensitivity and specificity for the models were: BOADICEA 0.81 (95% CI: 0.77-0.84), 26.4% and 98.8%; IBIS 0.80 (95% CI: 0.77-0.84), 19.4% and 97.3%; and Gail 0.76 (95% CI: 0.73-0.80), 17.3% and 98.5%. Conclusions: The breast cancer risk prediction models performed similarly although on closer appraisal, the BOADICEA and IBIS models performed slightly better. These models are simple, appropriate, cost-effective, and non-invasive tools for identifying high-risk women in low- and middle-income countries who could benefit from mammography screening.

Identification of genetic biomarkers in urine for early detection of prostate cancer.

Prostate cancer screening is a challenging and vital issue in the aspects of the current tests and risk assessments. Prostate cancer risk assessments are currently carried out by using blood, urine and tissue biomarkers with radiological imaging methods. Here, we introduce a novel noninvasive screening tool for a further in-depth selection of eligible cases for prostate biopsies which is based on sequencing somatic and hereditary HOXB13 mutations in urine samples. This approach provides diagnostic information to the physician about the presence of prostate cancer while aiming to screen for specific prostate biopsies and save biopsies potentially when there are no mutations related to prostate cancer. Findings suggest that this method is reliable, cost-effective, and has a promising potential in prostate cancer screening.

Risk Prediction Model Development for Late On-Set Breast Cancer Screening in Low- and Middle-Income Societies: A Model Study for North Cyprus.

Background: Early detection of breast cancer alters the prognosis and tools that can predict the risk for breast cancer in women will have a significant impact on healthcare systems in low- and middle-income regions, such as North Cyprus. Objective: In this study, we developed a simple breast cancer risk model for the women of North Cyprus. Methods: Data from 655 women, consisting of 318 breast cancer cases and 337 hospital-based controls, was used to develop and internally validate the model, external validation was carried out using, 653 women consisting of 126 cases and 527 controls. Data were obtained from medical records and interviews after informed consent. Results: A model was derived that consisted of age ≥50 years and <50 years and the presence and absence of >1 first-degree relatives (FDR) with breast cancer. From internal and external validations the model's AUCs were, 0.66 (95% CI = 0.62-0.70) and 0.69 (95% CI = 0.63-0.74) respectively. Conclusions: A unique model for risk prediction of breast cancer was developed to aid in identifying high-risk women from North Cyprus that can benefit from mammogram screening. Further study on a large scale that includes environmental risk factors is warranted.

Impact of circadian disruption on health; SIRT1 and Telomeres.

Circadian clock is a biochemical oscillator in organisms that regulates the circadian rhythm of numerous genes over 24 h. The circadian clock is involved in telomere homeostasis by regulating the diurnal rhythms of telomerase activity, TERT mRNA level, TERRA expression, and telomeric heterochromatin formation. Particularly, CLOCK and BMAL1 deficiency contribute to telomere shortening by preventing rhythmic telomerase activity and TERRA expression, respectively. Telomere shortening increases the number of senescent cells with impaired circadian rhythms. In return, telomerase reconstitution improves impaired circadian rhythms of senescent cells. SIRT1 that is an NAD+-dependent deacetylase positively regulates circadian clock and telomere homeostasis. SIRT1 contributes to the circadian clock by mediating CLOCK/BMAL1 complex formation, BMAL1 transcription and PER2 disruption. On the other hand, SIRT1 ensures telomere homeostasis by inducing telomerase and shelterin protein expression and regulating telomere heterochromatin formation. SIRT1 inhibition leads to both circadian clock and telomeres dysfunction that inhibit its activity. In light of this current evidence, we could suggest that the BMAL1/CLOCK complex regulates the telomere homeostasis in SIRT1 dependent manner, and also telomere dysfunction inhibits circadian clock function by suppressing SIRT1 activity to induce age-related diseases. We consider that increasing SIRT1 activity can prevent age-related diseases and help healthy aging by protecting telomere integrity and circadian clock function for individuals subjected to circadian rhythm disruption such as shift works, individuals with sleep disorders, and in the elderly population.

Updated North Cyprus response status for COVID-19 in comparison with similar country sizes. Highlights on the importance of population per square meter.

The COVID-19 pandemic is straining health systems worldwide. The World Health Organization has provided guidelines on a set of targeted and immediate actions that countries can use on a national, regional, and local level. Recommendations go from public hand hygiene stations, making face masks use obligatory, testing, and index finding which together with national closure of borders have been used to limit the disease so the countries' health care system can cope with the challenges. This is especially important as there seems to be an increased mortality rate even in countries normally regarded as well-functioning and having strong health system. In more susceptible countries this maybe even more important. The effect of these measures should be easier seen in small communities or countries. Societies have acted differently on when to apply the lockdown but most European countries have initiated lockdown after the first SARS-CoV-2 was diagnosed in their countries. With the COVID-19 pandemic and its economic consequences, it became especially important to re-evaluate the effect of response in light of a possible second wave. Yet, little is known about the effect of lockdown with respect to disease development and its handling. Hereby, we compare responses from relatively small 17 European countries including islands in three groups based on their population and report the response from North Cyprus in comparison to other small European countries. Our results indicate the importance of population per meter square, degree of isolation from others as well as social distancing, hygiene rules, timing of lockdowns in response to COVID-19 pandemic in small countries/ societies that are more susceptible for overwhelming their health system.

Tissue-Specific Ultra-Short Telomeres in Chronic Obstructive Pulmonary Disease.

Purpose: Telomere biology, especially tissue-specific ultra-short telomeres, might provide a strong contribution to our current knowledge in COPD development as well as a predictive marker for prognosis. To test this hypothesis, we investigated telomere lengths in lung tissue and leukocytes in patients diagnosed with COPD. Patients and Methods: Thirty-two patients were included in the current study. All patients showed a post-bronchodilator ratio of less than 70% post-bronchodilator predicted value of forced expiratory volume in second (FEV1%), mean 56%; range [19% to 86%]. To be able to investigate ultra-short telomeres, universal single telomere length analysis (U-STELA) was used. Results: Our results showed a higher level of the ultra-short telomere presence in bronchoalveolar lavage (BAL) cells when compared to leukocytes with statistical significance t(62)=5.771, p<0.00001. The FEV1% was lower in subjects with ultra-short telomeres in BAL (50.6% vs 81.6%: p<0.001) and in ultra-short telomeres in blood leukocytes (37.3% vs 58.5%: p=0.051) when compared to subjects without ultra-short telomeres in leukocytes. Furthermore, the patients who had ultra-short telomeres in BAL samples were significantly older (p=0.014) than patients who did not have ultra-short telomeres. Ultra-short telomeres in BAL (p=0.05) but not in leukocytes (p=0.33) were associated with FEV1% in a regressions model adjusting for age (p<0.0001), ever smoking (p<0.0001) and sex (p=0.71). The patients with ultra-short telomeres were graded higher in the Global Initiative for Chronic Obstructive Lung Disease (GOLD) classification (p=0.006). Conclusion: This study emphasizes the need to investigate the correct tissue to get a representative evaluation of the stage or advancedness of COPD. To our knowledge, this is the first study to show that there is a correlation between the presence of ultra-short telomeres in lung tissue and COPD severity. Our results suggest that ultra-short telomeres are involved in the molecular pathogenesis of COPD and might be used as a tissue-specific predictive biomarker.

A Solid Ultra Fine Self-Nanoemulsifying Drug Delivery System (S-SNEDDS) of Deferasirox for Improved Solubility: Optimization, Characterization, and In Vitro Cytotoxicity Studies.

The research work was designed to develop a solid self-nanoemulsifying drug delivery system (S-SNEDDS) of deferasirox (DFX). According to the solubility studies of DFX in different components, Peceol, Kolliphor EL, and Transcutol were selected as excipients. Pseudo-ternary phase diagrams were constructed, and then SNEDDS formation assessment studies and solubility of DFX in selected SNEDDSs formulations were performed. DFX loaded SNEDDS were prepared and characterized. The optimum DFX-SNEDDS formulations were developed. The relative safety of the optimized SNEDDS formulation was examined in a human immortalized myelogenous leukemia cell line, K562 cells, using the MTT cell viability test. Cytotoxicity studies revealed more cell viability (71.44%) of DFX loaded SNEDDS compared to pure DFX (3.99%) at 40 µM. The selected DFX-SNEDDS formulation was converted into S-SNEDDS by adsorbing into porous carriers, in order to study its dissolution behavior. The in vitro drug release studies indicated that DFX release (Q5%) from S-SNEDDS solidified with Neusilin UFL2 was significantly higher (93.6 ± 0.7% within 5 min) compared with the marketed product (81.65 ± 2.10%). The overall results indicated that the S-SNEDDS formulation of DFX could have the potential to enhance the solubility of DFX, which would in turn have the potential to improve its oral bioavailability as a safe novel delivery system.

Use of U-STELA for Accurate Measurement of Extremely Short Telomeres.

Telomeres are repetitive genetic materials that protect the chromosomes by capping the ends of chromosomes. Each time a cell divides, telomeres get shorter. Telomere length is mainly maintained by telomerase. This enzyme is present in the embryonic stem cells in high concentrations and declines with age. It is still unclear to what extend there is telomerase in adult stem cells, but considering these are the founder cells to the cells of the all tissues in a body, understanding the telomere dynamics and expression of telomerase in adult stem cells is very important.Telomere length has been implicated as one of the markers for neoplastic transformation in both in vivo and in vitro studies. During cancerogenesis, telomeres shorten due to high cell turnover and repeats are added by active telomerase or alternative lengthening of telomeres (ALT). This gradual shortening is replication driven and does not necessarily explain the presence of ultrashort telomeres. Ultrashort telomeres are observed when there is a sudden shortening in telomeres not related with cell division and may arise from breaks in telomeres due to oxidative damage and replication slippage.Universal STELA is an accurate method for evaluation of ultrashort telomeres in hMSC-telo1 cells. Compared to TRF assay, U-STELA is developed to overcome several problems in detecting abrupt telomere shortening in a single chromosome.

Role of TRF2 and TPP1 regulation in idiopathic recurrent pregnancy loss.

Telomeres are the tandem repeats (TTAGGG) present at the ends of the chromosomes that ensure chromosome stability and protect chromosomes from degradation. Telomeres in somatic human cells shorten after every cellular division and are linked to the cellular senescence. In this study we have investigated telomere length and expression of shelterin genes in aborted fetus material from idiopathic recurrent pregnancy losses. Telomere length was measured using Telomere Restriction Fragment Length (TRF) analysis. The gene expression levels for important shelterin complex proteins (TRF1, TRF2, POT1, and TPP1) were determined by Real-time Quantitative Reverse Transcriptase PCR (qRT-PCR). Our results demonstrated down regulation of TRF2 and TPP1 and a strong decline in average telomere length in abort material from women suffering from idiopathic recurrent pregnancy loss. We suggest that shorter telomere length and downregulation of the major shelterin components TRF2 and TPP1 leading to "telomere uncapping", might play a critical role in recurrent pregnancy loss.

Telomere stability and telomerase in mesenchymal stem cells.

Telomeres are repetitive genetic material that cap and thereby protect the ends of chromosomes. Each time a cell divides, telomeres get shorter. Telomere length is mainly maintained by telomerase. This enzyme is present in high concentrations in the embryonic stem cells and in fast growing embryonic cells, and declines with age. It is still unclear to what extent there is telomerase in adult stem cells, but since these are the founder cells of cells of all the tissues in the body, understanding the telomere dynamics and expression of telomerase in adult stem cells is very important. In the present communication we focus on telomere expression and telomere length in stem cells, with a special focus on mesenchymal stem cells. We consider different mechanisms by which stem cells can maintain telomeres and also focus on the dynamics of telomere length in mesenchymal stem cells, both the overall telomere length and the telomere length of individual chromosomes.