Effect of Pulsed Low-Level Lasers on Adult versus Neonatal Human Retinal Pigment Epithelial Cells: An in-vitro Study.

Purpose: To investigate the short-term effects of low-level lasers (LLLs; also known as low-power laser therapy) on the structure, genetic, and phenotype of cultured human retinal pigment epithelial (hRPE) cells from both adult and neonatal sources. Methods: Cultivated adult and neonatal hRPE cells were irradiated with two types of LLL (630 nm and 780 nm), 1 min daily for five consecutive days. Results: An increase in doubling time was observed in 630 nm-irradiated adult hRPE cells (P = 0.032). The gene expression profile revealed increased expression of retinoid isomerohydrolase RPE65 (RPE65) (P < 0.01 for 630 nm laser, P < 0.001 for 780 nm laser) and nestin (NES) (P < 0.01 for 630 nm laser) in neonatal hRPE cells, upregulation of RPE65 (P < 0.001 for 780 nm laser) and paired box 6 (PAX6) (P < 0.001 for 780 nm laser) genes in adult hRPE cells, and reduced expression of actin alpha 2 (ACTA2) in 780 nm-irradiated adult hRPE cells (P < 0.001). Except the significant increase of α -SMA in 780 nm-irradiated neonatal hRPE cells, no significant change was noted in the expressions of other investigated proteins. Conclusion: Short-term irradiation of neonatal and adult hRPE cells with LLLs may induce multipotency at the transcriptional level. Irradiation of neonatal hRPE cells with LLLs can be associated with increased risk of myofibroblastic transformation; however, adult hRPE cells irradiated with the 780 nm laser have minimal risk of myofibroblastic differentiation. It seems that the 780 nm laser may be a promising option for future photobiomodulation in retinal degenerations in adults.

Biological feasibility of discharge a local WTTP sludge to sewer network and centralized WWTP; a case study: Tehran, Iran.

Over the recent years, ever-increasing population growth and higher wastewater production has been a challenge for decentralized wastewater treatment plants (WWTPs). In addition, sludge treatment due to high cost for equipment and place make authorities to find a sustainable approach in both of economical and technical perspectives. One of the proposed solutions is transferring the sludge produced from decentralized WWTP to centralized WWTP. However, the appropriate proportional ratio of raw sludge to raw sewage is a challenge, otherwise, it make anaerobic conditions and sewage rotting along the sewer network based on permissible limit of dihydrogen sulfide (H2S) gas (5 ppm). In the present study, seven reactors with different ratios of sludge to raw sewage (0, 15, 20, 25, 50, 75, 100) were used to stimulate the feasibility of transferring Shahrake Gharb WWTP sludge along the wastewater transfer pipe to the centralized sewage treatment south Tehran WWTP plant in Tehran, Iran. The septic situation and H2S emission of different reactors within 7 h (Time to reach the compound in the south treatment plant) was analyzed by gas meter. The results indicated that the optimum ratio of sludge to raw sewage was 15% without H2S production during 7 h. In addition, due to the high volume of sludge produced by the Shahrake Gharb WWTP, the optimal ratio of lime to total solids (TS) in sludge (gr/gr) (0.6) increased the sludge loading rate from 15 to 30% without any H2S emission during the stimulation study period. Therefore, the lime stabilization and transfer of sludge from a decentralized WWTP to a centralized WWTP is a feasible way to manage the sludge and enhance the treatment capacity in local WWTP.

Biomaterials used for tissue engineering of barrier-forming cell monolayers in the eye.

Cell monolayers that form a barrier between two structures play an important role for the maintenance of tissue functionality. In the anterior portion of the eye, the corneal endothelium forms a barrier that controls fluid exchange between the aqueous humor of the anterior chamber and the corneal stroma. This monolayer is central in the pathogenesis of Fuchs endothelial corneal dystrophy (FECD). FECD is a common corneal disease, in which corneal endothelial cells deposit extracellular matrix that increases the thickness of its basal membrane (Descemet's membrane), and forms excrescences (guttae). With time, there is a decrease in endothelial cell density that generates vision loss. Transplantation of a monolayer of healthy corneal endothelial cells on a Descemet membrane substitute could become an interesting alternative for the treatment of this pathology. In the back of the eye, the retinal pigment epithelium (RPE) forms the blood-retinal barrier, controlling fluid exchange between the choriocapillaris and the photoreceptors of the outer retina. In the retinal disease dry age-related macular degeneration (dry AMD), deposits (drusen) form between the RPE and its basal membrane (Bruch's membrane). These deposits hinder fluid exchange, resulting in progressive RPE cell death, which in turn generates photoreceptor cell death, and vision loss. Transplantation of a RPE monolayer on a Bruch's membrane/choroidal stromal substitute to replace the RPE before photoreceptor cell death could become a treatment alternative for this eye disease. This review will present the different biomaterials that are proposed for the engineering of a monolayer of corneal endothelium for the treatment of FECD, and a RPE monolayer for the treatment of dry AMD.

In-line leukoreduction filters: A new source of microparticle for human and animal study.

INTRODUCTION: The isolation of microparticles (MPs) from leukoreduction filters (LRFs) during cell extraction process introduced LRFs as a precious source of MPs for animal and human study. METHOD: LRFs were collected from Tehran Blood Transfusion Center. The back-flushing method was used for leukocyte extraction from the LRFs. MPs were isolated through double-step centrifugation. Dynamic light scattering (DLS), electron microscopy (EM), and flow cytometry were performed for the evaluation of MPs size, morphology, and structural properties respectively. Statistical analyses were carried out to evaluation of differences between test and control groups. a p-value less than 0.05 indicates significant differences. RESULT: DLS analysis showed that the average MP size in the test and control groups was 654.83 nm and 233.68 nm respectively. SEM images showed the spherical, oval, cell fragment, and micro-aggregate particles and TEM images demonstrated the mitochondrial-like body in the MPs. Flow cytometry studies also showed a significant increase in the percent of CD41, and CD14, and a significant decrease in the percent of CD235a in the test group compared to control (P value=0.029, P value=0.035, P value= 0.001 respectively). Moreover, the percentage of CD34 MPs indicated a borderline difference between the two groups (P value= 0.075). Finally count of MPs in the test and control groups was 1202095.34 and 280948.64, respectively and the difference was significant (P value=0.008). CONCLUSION: It is concluded that LRFs are a potential source of the large volume of various cell MPs with different phenotypical and structural properties for animal and human phase studies. Moreover, the investigation of LRFs as a source of different types of exosomes can shed new light on extracellular vesicle studies.

Highlighting the interaction between immunomodulatory properties of mesenchymal stem cells and signaling pathways contribute to Graft Versus Host Disease management.

Background Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) has been increasingly used as a therapeutic approach for hematological malignancies. Several potential strategies have been developed for treating or preventing allo-HSCT complications, specifically graft-versus-host disease (GVHD). GVHD could significantly affect the morbidity and mortality of patients after allo-HSCT. Curative treatment and prophylaxis regimens for GVHD could reduce GVHD incidence and improve survival rate. Among these therapeutic strategies, mesenchymal stem cell (MSCs) mediated immunomodulation has been explored widely in clinical trials. MSCs immunomodulation ability in GVHD correlates with the interactions of MSCs with innate and adaptive immune cells. However, signaling pathways responsible for MSCs' impact on GVHD regulation, like JAK/STAT, NOTCH, MAPK/ERK, and NFκß signaling pathways, have not been clearly described yet. This review aims to illuminate the effect of MSCs-mediated immunomodulation in GVHD management after allo-HSCT representing the role of MSCs therapy on signaling pathways in GVHD. Conclusion MSCs could potentially modulate immune responses, prevent GVHD, and improve survival after allo-HSCT. Previous studies have investigated different signaling pathways' contributions to MSCs immunoregulatory ability. Accordingly, targeting signaling pathways components involved in MSCs related GVHD regulation is proven to be beneficial.

The association of disease type, pre-transplant hemoglobin level and platelet count with transfusion requirement after autologous hematopoietic stem cell transplantation.

BACKGROUND: Autologous hematopoietic stem cell transplantation (auto-HSCT) has become an effective treatment for a wide range of hematologic and non-hematologic diseases. Patients undergoing HSCT might require multiple platelets and red blood cell (RBC) transfusions during aplasia phase until engraftment, which could profoundly affect patients' conditions. Identification of risk factors associated with blood product requirements could help in decreasing transfusion-related complications. We evaluated the association of disease type, pre-transplant hemoglobin level, and pre-transplant platelet count with RBC/platelet transfusion requirement after auto-HSCT. METHODS: In this retrospective study, 324 patients diagnosed with multiple myeloma (MM), Hodgkin disease (HD), and non-Hodgkin lymphoma (NHL) and underwent auto-HSCT were included. The associations of disease type, pre-transplant hemoglobin level, and platelet count with post-transplant packed cell and single-/random-donor platelet transfusions were evaluated. RESULTS: Our study results illustrated that the higher pre-transplant hemoglobin level significantly decreased the post-HSCT requirement for packed cell (IRR=0.81, [CI: 9.73-0.90], P=0.0001), while the pre-transplant platelet showed no significant relationship with platelet requirement after HSCT. HD was associated with increment in packed cell (IRR=2.04, [CI: 1.35-3.08], P=0.001) and single donor platelet (IRR=1.39, [CI: 1.09-1.78], P=0.008) requirement after transplant. The trends showed that a higher platelet level led to a lower need for platelet transfusion. CONCLUSION: Pre-transplant hemoglobin level could be valuable markers for predicting post-HSCT RBC requirements and might be beneficial for better management of transfusion requirements to minimize the transfusion-related complications. Patients with HD seem to be more prone to blood product requirements post-transplant.

In-depth summary over cytomegalovirus infection in allogeneic hematopoietic stem cell transplantation recipients.

In this study, we reviewed various aspects of cytomegalovirus infection, including pathophysiology, diagnosis methods, and antiviral treatments. Background: Infections continue to be a major reason of complications like high non-relapse morbidity and mortality rate after allogenic hematopoietic stem cell transplantation. Cytomegalovirus is the most common infection in immunocompromised patients or those with graft-versus-host disease. The Latent-cytomegalovirus disease could increase the risk of reactivation in allogenic hematopoietic stem cell transplantation patients and lead to profound adverse effects on transplantation outcomes. Cytomegalovirus-specific CD4 + and CD8 + T cells reconstitution is crucial for protection against the virus reactivation. Different prophylactic, pre-emptive, and therapeutic anti-viral drugs are available to prevent cytomegalovirus infection/reactivation and treat resistant infections. Conclusion: Although there has been introduced various CMV antiviral treatment strategies like antiviral drugs, Vaccination, passive immunotherapies and adoptive transfer of CMV-specific T cells, further clinical trials are required to approve current therapies.

Optimizing the effective doses of mitomycin C, 5-fluorouracil, and their combination on cultivated basal cell carcinoma / Otimização da dose efetiva da mitomicina C, do 5fluorouracil e da combinação de ambos em culturas de células de carcinoma basocelular

ABSTRACT Purpose: This study aimed to optimize the effective doses of mitomycin C, 5-fluorouracil, and their combination on cultivated basal cell carcinoma. Methods: Cultivated basal cell carcinoma and fibroblastic cells were treated with different concentrations of mitomycin C, 5-fluorouracil, and their combination. Cell viability, cell cycle, apoptosis, and expression levels of TP53, CDKN1A, and CDK6 were investigated. The most effective drug with its optimum dosage was administered via multiple intralesional injections to a 65-year-old woman with advanced periorbital nodulo-ulcerative BCC. Results: The concentrations of 0.00312 and 0.312 mg/mL were considered optimum for mitomycin C and 5-fluorouracil, respectively. The mean viabilities of basal cell carcinoma treated with mitomycin C alone and its combination with 5-fluorouracil were significantly less than those of the controls (p=0.002 and p=0.04, respectively). The cell cycle of all the treated basal cell carcinoma groups was arrested in the S phase. The apoptotic rates (p=0.002) of mitomycin C treated basal cell carcinoma were higher than those of the other treated cells, and their TP53 was significantly upregulated (p=0.0001). Moreover, CDKN1A was upregulated, whereas CDK6 was downregulated in basal cell carcinoma treated with either 5-fluorouracil (p=0.0001 and p=0.01, respectively) or the combination of 5-fluorouracil and mitomycin C (p=0.007 and p=0.001, respectively). Basal cell carcinoma lesions were significantly alleviated following mitomycin C injections in the reported patient. Conclusion: Our in vitro results revealed that the effective doses of mitomycin C and 5-fluorouracil on cultivated basal cell carcinoma were optimized. Mitomycin C was more effective in inducing the apoptosis of basal cell carcinoma than 5-fluorouracil and their combination. The intralesional injections of the optimum dose of mitomycin C could be proposed for the nonsurgical treatment of advanced eyelid basal cell carcinoma.

RESUMO Objetivo: Otimizar a dose efetiva de mitomicina C, 5fluorouracil e da combinação de ambos em culturas de células de carcinoma basocelular (CBC). Métodos: Culturas de células de células de carcinoma basocelular e de fibroblastos foram tratadas com diferentes concentrações de mitomicina C, 5fluorouracil e combinação de ambos. Além disto, foram investigados a viabilidade celular, o ciclo celular, a apoptose e a expressão dos genes TP53, CDKN1A e CDK6. O medicamento mais eficaz, em sua dosagem otimizada, foi administrado em últiplas injeções intralesionais em uma mulher de 65 anos com carcinoma basocelular nódulo-ulcerativo periorbital avançado. Resultados: A concentração de 0,00312 mg/mL de mitomicina C e a de 0,312 mg/mL de 5fluorouracil foram consideradas as ideias. A viabilidade média das células de carcinoma basocelular tratadas com mitomicina C isoladamente e em combinação foi significativamente menor que nas células de controle (respectivamente, p=0,002 e p=0,04). Todos os grupos de carcinoma basocelular tratados demonstraram interrupção do ciclo celular na fase S. As células de carcinoma basocelular tratadas com mitomicina C mostraram maiores taxas de apoptose (p=0,002) e significativa regulação positiva do gene TP53 (p=0,0001). Além disso, o gene CDKN1A foi positivamente regulado e o gene CDK6 foi negativamente regulado em células de carcinoma basocelular tratadas com 5fluorouracil (respectivamente, p=0,0001 e p=0,01) ou com a combinação de medicamentos (respectivamente, p=0,007 e p=0,001). Injeções posteriores de mitomicina C na paciente em questão levaram à melhora significativa da lesão do carcinoma basocelular. Conclusão: Nossos resultados in vitro otimizaram as doses efetivas de mitomicina C e 5fluorouracil na cultura de células de carcinoma basocelular e mostraram que a mitomicina C tem mais eficácia na apoptose de células de carcinoma basocelular do que o 5fluorouracil e a combinação de ambos. Injeções intralesionais de doses otimizadas de mitomicina C podem ser propostas para o tratamento não cirúrgico do células de carcinoma basocelular avançado de pálpebra.

Immunotherapy with adoptive cytomegalovirus-specific T cells transfer: Summarizing latest gene engineering techniques.

Cytomegalovirus (CMV) infection remains a major complication following allogeneic hematopoietic stem cell transplantation (HSCT). T cell response plays a critical role in inducing long-term immunity against CMV infection/reactivation that impairs during HSCT. Adoptive T cell therapy (ACT) via transferring CMV-specific T cells from a seropositive donor to the recipient can accelerate virus-specific immune reconstitution. ACT, as an alternative approach, can restore protective antiviral T cell immunity in patients. Different manufacturing protocols have been introduced to isolate and expand specific T cells for the ACT clinical setting. Nevertheless, HLA restriction, long-term manufacturing process, risk of alloreactivity, and CMV seropositive donor availability have limited ACT broad applicability. Genetic engineering has developed new strategies to produce TCR-modified T cells for diagnosis, prevention, and treatment of infectious disease. In this review, we presented current strategies required for ACT in posttransplant CMV infection. We also introduced novel gene-modified T cell discoveries in the context of ACT for CMV infection. It seems that these innovations are enabling to improvement and development of ACT utilization to combat posttransplant CMV infection.

Iranian researchers' contributions to research on COVID-19: A bibliometric analysis and visualization.

Background: COVID-19 pandemic crisis motivated researchers worldwide to deeply investigate it from different perspectives. As Iran is one of the highly-affected countries by Covid-19, Iranian researchers have focused on studying it. This study aimed at analyzing and visualizing Iranian researchers' papers on COVID-19 from a bibliometric perspective. Methods: By searching MeSH-selected keywords related to COVID-19 in Scopus, Iranian researchers' papers on COVID-19 were extracted in a CSV format and underwent bibliometric techniques, such as coauthorship analysis, citation, and co-citation analysis, keyword and term co-occurrence mapping and etc. in the Microsoft Excel and VOSviewer software package. Results: A total of 405 papers were authored by Iranian researchers on COVID-19 during the study period, with the average number of citations per paper of 2.60 and a mean h-index of 15. The majority of papers were original articles in English. Archives of Clinical Infectious Diseases and Archives of Iranian Medicine and Medical Hypotheses were highly ranked publishing journals, respectively. The most productive institute and author were Tehran University of Medical Sciences with 119 papers and Rezaei, N. with 12 papers. Iranian researchers collaborated with the researchers of 73 countries, with the USA ranking first in Covid-19 research, followed by Italy, Canada, and United Kingdom. In publishing papers on COVID-19, Iran ranked first among the Middle Eastern countries and thirteenth internationally. Conclusion: Iranian researchers were active in 5 main areas of COVID-19 research, including epidemiology, diagnosis, treatment, virology, and systematic review.

The apheresis content analysis in Allo-HSCT represents reliable influential factors on graft-versus-host disease and overall survival.

BACKGROUND: Hematopoietic stem cell transplantation (HSCT) is an established treatment for hematologic malignancies. However the post-HSCT outcome can be affected by multiple pre-transplant, transplant, and post-transplant factors. The cellular content of graft could be possible factors influencing the graft-versus-host disease (GVHD) and overall survival (OS) as transplantation outcomes. PURPOSE: The aim of this study was to assess the impact of infused CD34+ cells, CD3+ cells, and MNC count on the patients' survival and incidence of graft-versus-host disease (GVHD). MATERIAL AND METHODS: We analyzed 87 patients with hematological malignancies who underwent allogeneic hematopoietic stem cell transplantation at the Taleghani Stem Cell Transplantation and Cell therapy center, Tehran, Iran from January 2016 to December 2018. Patients were conditioned with either myeloablative conditioning regimen or reduced-intensity regimen. RESULT: A CD34+ cell dose < 4.35 × 106/kg and CD3+ cell dose < 365 × 106/kg was associated with higher survival and lower acute and chronic GVHD incidence, although their association was not statistically significant. Moreover, there was a significant association between MNC count < 6.15 × 108/kg and acute GVHD incidence. CONCLUSION: Graft cell dose, lower than the cut-off level, could lead to better outcomes after allogeneic transplantation. However, this study showed that future investigations are required in a larger population of patients in order to determine the exact effect of allogeneic graft cell dose on transplantation outcome.

Optimizing the effective doses of mitomycin C, 5-fluorouracil, and their combination on cultivated basal cell carcinoma.

PURPOSE: This study aimed to optimize the effective doses of mitomycin C, 5-fluorouracil, and their combination on cultivated basal cell carcinoma. METHODS: Cultivated basal cell carcinoma and fibroblastic cells were treated with different concentrations of mitomycin C, 5-fluorouracil, and their combination. Cell viability, cell cycle, apoptosis, and expression levels of TP53, CDKN1A, and CDK6 were investigated. The most effective drug with its optimum dosage was administered via multiple intralesional injections to a 65-year-old woman with advanced periorbital nodulo-ulcerative BCC. RESULTS: The concentrations of 0.00312 and 0.312 mg/mL were considered optimum for mitomycin C and 5-fluorouracil, respectively. The mean viabilities of basal cell carcinoma treated with mitomycin C alone and its combination with 5-fluorouracil were significantly less than those of the controls (p=0.002 and p=0.04, respectively). The cell cycle of all the treated basal cell carcinoma groups was arrested in the S phase. The apoptotic rates (p=0.002) of mitomycin C treated basal cell carcinoma were higher than those of the other treated cells, and their TP53 was significantly upregulated (p=0.0001). Moreover, CDKN1A was upregulated, whereas CDK6 was downregulated in basal cell carcinoma treated with either 5-fluorouracil (p=0.0001 and p=0.01, respectively) or the combination of 5-fluorouracil and mitomycin C (p=0.007 and p=0.001, respectively). Basal cell carcinoma lesions were significantly alleviated following mitomycin C injections in the reported patient. CONCLUSION: Our in vitro results revealed that the effective doses of mitomycin C and 5-fluorouracil on cultivated basal cell carcinoma were optimized. Mitomycin C was more effective in inducing the apoptosis of basal cell carcinoma than 5-fluorouracil and their combination. The intralesional injections of the optimum dose of mitomycin C could be proposed for the nonsurgical treatment of advanced eyelid basal cell carcinoma.

Cancer Care Management During the COVID-19 Pandemic.

New cases of the novel coronavirus, also known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are increasing around the world. Currently, health care services are mainly focused on responding to and controlling the unique challenges of the coronavirus disease 2019 (COVID-19) pandemic. These changes, along with the higher susceptibility of patients with cancer to infections, have profound effects on other critical aspects of care and pose a serious challenge for the treatment of such patients. During the COVID-19 pandemic, it is important to provide strategies for managing the treatment of patients with cancer to limit COVID-19-associated risks at this difficult time. The present study set out to summarize the latest research on epidemiology, pathogenesis, and clinical features of COVID-19. We also address some of the current challenges associated with the management of patients with cancer during the COVID-19 pandemic and provide practical guidance to clinically deal with these challenges.

Effects of fibrin glue as a three-dimensional scaffold in cultivated adult human retinal pigment epithelial cells.

This study was conducted to examine morphological, genotypic, and phenotypic alterations occurring in cultured adult human retinal pigment epithelial cells when encapsulated with different concentrations of fibrin glue. Cultivated adult human retinal pigment epithelial cells were encapsulated with different concentrations of fibrin glue, namely FG1 (42 mg/dl), FG2 (84 mg/dl), FG3 (124 mg/dl), FG4 (210 mg/dl), followed by the evaluation of genetic and cytomorphological changes and protein expression. Cultured adult human retinal pigment epithelial cells showed dendritiform morphology during the early days of encapsulation with fibrin glue. Moreover, an increasing inhibitory effect on cell growth was observed with increasing concentrations of fibrin glue. At the transcriptional level, the expression of MMP2, PAX6, and ITGB1 in FG1-encapsulated cells was significantly higher than that in other treated groups; however, the expression of ACTA2 was lower in all fibrin glue-encapsulated groups compared to that in the controls. Immunocytochemistry showed that FG2-encapsulated cells expressed cytokeratin 8/18, RPE65, and ZO-1 proteins, but not PAX6. In conclusion, fibrin glue at a concentration of 84 mg/dl allows proper encapsulation of adult human retinal pigment epithelial cells, while preserving the morphometric, genotypic, and phenotypic features of the cells. This three-dimensional biopolymer can be considered a reliable vehicle for retinal pigment epithelium cell transplantation in cell-based therapies.