Application of methylene blue for the prevention and treatment of COVID-19: A narrative review.

The newest virus from the SARS family of viruses called acute syndrome-coronavirus-2 (SARS-CoV-2), which causes COVID-19 disease, was identified in China at the end of 2019. In March 2020, after it spread to 29 additional countries, it was declared a pandemic by the World Health Organization (WHO). SARS-CoV-2 infection mainly starts through the respiratory tract and causes a wide spectrum of symptoms from asymptomatic infections to acute respiratory distress syndrome with multi-organ failure and vasoplegic shock. Among the many immunomodulatory and antiviral drugs that have been studied for the treatment of COVID-19, methylene blue (MB) may play an influential role. This article reviews the history of MB applications, the antiviral effects of MB against SARS-CoV-2, and the results of in vivo and in vitro studies of the use of MB in COVID-19. Based on studies, MB can simultaneously affect most of the host's harmful responses caused by SARS-CoV-2 infection due to its multiple properties, including anti-hypoxemia, anti-oxidant, immune system modulator, and antiviral. The use of MB is associated with a reduction in the possibility of getting infection, and mortality, and can be used as a safe, effective, cheap, and available treatment option with minimal side effects for the clinical management of COVID-19.

Treatment of recalcitrant esophagocutaneous fistula by platelet-rich-plasma-fibrin glue: A case study.

INTRODUCTION: Recalcitrant esophagocutaneous fistula is a very uncommon complication after neck surgery. Management of this non-healing fistula has long been a topic of debate. This report provides an approach for treating it. PRESENTATION OF CASE: A 65-year-old woman presented nineteen years after branchial cleft cyst surgery with cyst recurrence associated with swelling. Sonography displayed a collection in the front of the left carotid artery in the suprasternal notch. On the CT, a similar finding was seen, a collection with gaseous density in front of the left sternocleidomastoid and a hypodense nodule on the right lobe of the thyroid. The pathology report describes an abscess with many macrophages and neutrophils, fat necrosis, microcalcification, and foreign body reaction around amorphous bodies. Again, surgical resection of the swelling area and tract was done and an esophago-cutaneous fistula was developed after surgery which did not heal after 6 months. CLINICAL DISCUSSION: According to accelerating the healing time and complete closure of chronic wounds such as lower-extremity diabetic ulcers, persistent pneumothorax, anal fistula, and recalcitrant gastrocutaneous fistula by using platelet-rich plasma (PRP) and fibrin glue (FG); PRP-FG can be considered as a safe and effective treatment option for chronic wound healing. So, for treatment of this fistula, PRP-FG was used. PRP-FG was obtained from the patient's blood and injected into the fistula tract. The discharge was stopped after one week and the fistula was cured. CONCLUSION: PRP-FG injection into the fistula tract provides a simple and non-invasive approach for the treatment of recalcitrant esophagocutaneous fistula.

Complete Salvage of Third-degree Burn of Ear by Repairing Gel-PRP-Fibrin Glue, A Case Report.

The ear is a very important facial feature and enhances the overall look of the face. Usually, patients with entire ear burns also have large areas of burn in their bodies. To save the patient, skin grafting to reduce the injured body surface area is prioritized and the treatment of the ears may be inevitably delayed, which results in minor to very severe cosmetic deformity. This has a deep effect on the life quality of a patient who survives a burn injury. Therefore, any step that decreases ear deformity or loss is very crucial for the patient. There are different challenging procedures for ear reconstruction which are very tedious works. In this study, for one patient, repairing gel-platelet-rich plasma-fibrin glue was applied for the treatment of a deep dermal burned ear as a novel, non-invasive, and simple approach to give a normal shape to the ear to prevent extensive morbidity during the acute phase and deformities later as a consequence of burns. No keloid is observed after treatment.

Persistent pneumothorax treatment following congenital cardiac surgery by platelet-fibrin glue.

Persistent pneumothorax is a life-threatening complication that can occur after congenital cardiac surgery. Traditional treatment such as chest tube drainage may not be effective in managing this condition. This study presents a new minimally invasive method for treating persistent pneumothorax using platelet-rich plasma-fibrin glue (PRP-FG). The method has been successful in treating postoperative chylothorax in previous studies, and its use has decreased morbidity, mortality, and hospital stay in chylothorax patients. Ten patients with persistent pneumothorax following cardiothoracic surgery (3 TAPVC, 2 d-TGA, 2 VSD + IAA,1 TRUNCUS + TAPVC, 1 VSD + COA, 1 GLENN), who did not respond to conservative management, underwent treatment with PRP-FG. Follow-up was done for a period of 1-4 years. The age and diagnoses of pneumothorax after surgery were 85.5 ± 36.0 days and 62.4 ± 34.3 h, respectively. Persistent pneumothorax of 8 patients (80%) was cured completely after PRP-FG injection. PRP-FG therapy was failed in two patients who died. All cured patients had a normal life without any complications during follow-up. After PRP-FG injection, 3 patients stopped bubbling at one-time injection, 3 patients stopped bubbling at two-time injection, and 2 patients stopped bubbling at three-time injection. Two patients died during treatment; in these cases, one-time injections were done which was not successful. Persistent pneumothorax after congenital-cardiac surgery can be treated successfully with PRP-FG. This bedside minimal-invasive procedure may significantly decrease the morbidity and mortality rate. Further research is needed to confirm the efficacy of this promising treatment through multicentre clinical trials.

Limb Saving with a Combination of Allogenic Platelets-Rich Plasma, Fibrin Glue, and Collagen Matrix in an Open Fracture of the Tibia: A Case Report.

Tibia open fractures and lower limb soft tissue injuries are undesired conditions for orthopedic and general surgeons, and there is no ideal procedure for treating recalcitrant ulcers or severe traumatic ulcers. Recently, several novel approaches have been proposed, such as bone marrow stem cells, platelets, fibrin glue, and collagen matrix. The current study was conducted to evaluate the effectiveness of combining platelets, fibrin glue, and a collagen matrix for treating a serious limb-threatening wound in a 33-year-old male with a right lower limb injury. After treatment, the wound was fully closed, tissue granulation was grown, skin grafting was done, and the right leg was saved. In conclusion, the combination of the mentioned components can be utilized to synergistically enhance wound healing and preserve the limb.

Autologous platelet-rich-plasma injection and platelet-rich fibrin glue interposition for treatment of anal fistula resistant to surgery.

Aim: The current study purposed to evaluate the autologous platelet-rich plasma (PRP), and platelet-rich fibrin glue (PRFG) effect on the treatment of complex, and recalcitrant anal fistula (AF) which was not cured by several surgeries. Background: AF has remained one of difficult challenges for centuries. Surgery is the common treatment method for it, but the risk of fecal incontinence and recurrence is still a distressing complication for patients and surgeons. New procedures were published in the scientific literature, each with advantages and disadvantages. According to reports, an effective therapy option is the autologous fibrin glue that is rich in platelets. Methods: Autologous PRP and PRFG were prepared from 10 patients' own blood. The surgeon curetted the tract of anal fistula for the deepithelialisation till hemorrhage occurred; PRP was injected around the fistula into the tissue, and PRFG was interpositioned in the tract. Age, number of previous surgeries, complications, number of PRP and PRFG administrations, and duration of halting the discharge were among the information gathered. Patients were followed up between 10 months to 84 months after treatment. Results: No complications were observed during and after the injection. During the period of follow-up, AF leakage was stopped for 6 patients, but not for 4 patients. Conclusion: Since autologous PRP injection, and PRFG interposition is a safe, effective, and minimally invasive procedure for resistant AF to surgeries; it can be used, along with surgery to increase the healing rate of complex anal fistula.

The Potential of Leucomethylene Blue in Methemoglobinemia Treatment: A New Hope for Patients with G6PD?

Methylene blue (MB) has been routinely used to treat methemoglobinemia. In the body, MB is reduced to leucomethylene blue (LMB) by NADPH-dependent methemoglobin (MetHB) reductase, and then LMB reduces Fe3+ to Fe2+. In glucose-6-phosphate dehydrogenase (G6PD) deficiency, NADPH is not produced sufficiently to protect erythrocytes against oxidative stress and to take part in relevant biochemical reactions. Since MB is an oxidative agent, its administration in individuals with G6PD deficiency leads to an increased risk of hemolysis through oxidative stress and even death. Therefore, its administration has been prohibited from treating methemoglobinemia in G6PD patients. As an antioxidant and direct reducing agent for Fe3+, LMB may be prescribed for treating MetHB in patients with G6PD deficiency. Considering the biochemical process of turning MB into LBM and the reducing nature of LMB, it seems LMB is a safer drug than MB in treating methemoglobinemia. LMB can even be administrated in other treatments without any concern about increasing oxidative stress, exacerbating the inflammation. Proof-of-concept experimental and clinical trials could substantiate this hypothesis.

The effects of grapes and their products on immune system: a review.

Immune system plays a significant role in preventing and controlling diseases. Some studies reported the beneficial effects of grapes and their products on immunity. However, their results are controversial. This review aimed to discuss the effects of grapes and their products on immune system and their mechanisms of action. Although various in-vio and in-vitro studies and some human studies suggested that grapes and their products may help to improve the immune system's function, clinical trials in this area are limited and inconsistent.In conclusions, although, consumption of grapes and their products may help to having a healthy immune syste, further studies particularly human studies are required to clarify the precise effects of them and their mechanisms regarding immune system.

Appropriate fixative for MEM-G/9 staining of cultured human HLA-G-positive JEG-3 trophoblast tumor cells.

Human leukocyte antigen (HLA-G) participates in immunosuppression and is useful for prenatal diagnostics. Isolation of fetal cells positive for HLA-G by HLA-G antibody conjugated nanoparticles from the cervix of pregnant women is the basis for non-invasive prenatal testing. Endocervical specimens are fixed in transport medium before isolation using antibody conjugated nanoparticles. Staining of HLA-G using MEM-G/9 antibody, however, is restricted to unfixed cells. We investigated the effect of several fixatives on the interaction of HLA-G with MEM-G/9 in the HLA-G-positive cell line, JEG-3. We investigated absolute methanol, 1:1 acetate buffer:methanol, Pap solution and paraformaldehyde. The effects of these fixatives were evaluated using immunofluorescence. We found no MEM-G/9 surface staining of methanol fixed cells. Approximately 40% of JEG-3 cells fixed with paraformaldehyde failed to stain. Nearly all cells were stained with MEM-G/9 following fixation with acetate buffer:methanol or Pap solution. Our findings indicate the importance of using an appropriate fixative for preserving HLA-G cell surface antigen for studies using the MEM-G/9 antibody.

Isolation of HLA-G+ cells using MEM-G/9 antibody-conjugated magnetic nanoparticles for prenatal screening: a reliable, fast and efficient method.

The development of an effective and noninvasive early method for obtaining fetal cells is crucial to prenatal screening. Despite proving the presence of fetal cells in the reproductive tract, their use is limited due to their inability to properly isolate them from maternal cells. Magnetic-activated cell sorting (MACS) is a simple technique to separate cells. The present study aimed to develop a MACS-based platform for the isolation of the HLA-G expressing trophoblast cells. For this purpose, first, the triazine functionalized MNPs were synthesized and characterized. Then, MNPs were directly and indirectly conjugated by the MEM-G/9 antibodies targeting HLA-G+ cells. The antibody amount on the surface of the nanoparticles was determined with the Bradford assay. The cell capture efficiency was also investigated. Various characterization methods confirmed the successful nanoparticle synthesis and antibody conjugation. The optimal initial antibody amount for the immobilization was about 20 µg and the optimal time was 3 h. The antibody-nanoparticles by the indirect method had better targeting and capture efficiency than the direct method. The MNPs indirectly conjugated with antibodies are an efficient tool for cell isolation and present considerable potential to be applied in biomedical fields.

Predicting the most deleterious missense nsSNPs of the protein isoforms of the human HLA-G gene and in silico evaluation of their structural and functional consequences.

BACKGROUND: The Human Leukocyte Antigen G (HLA-G) protein is an immune tolerogenic molecule with 7 isoforms. The change of expression level and some polymorphisms of the HLA-G gene are involved in various pathologies. Therefore, this study aimed to predict the most deleterious missense non-synonymous single nucleotide polymorphisms (nsSNPs) in HLA-G isoforms via in silico analyses and to examine structural and functional effects of the predicted nsSNPs on HLA-G isoforms. RESULTS: Out of 301 reported SNPs in dbSNP, 35 missense SNPs in isoform 1, 35 missense SNPs in isoform 5, 8 missense SNPs in all membrane-bound HLA-G isoforms and 8 missense SNPs in all soluble HLA-G isoforms were predicted as deleterious by all eight servers (SIFT, PROVEAN, PolyPhen-2, I-Mutant 3.0, SNPs&GO, PhD-SNP, SNAP2, and MUpro). The Structural and functional effects of the predicted nsSNPs on HLA-G isoforms were determined by MutPred2 and HOPE servers, respectively. Consurf analyses showed that the majority of the predicted nsSNPs occur in conserved sites. I-TASSER and Chimera were used for modeling of the predicted nsSNPs. rs182801644 and rs771111444 were related to creating functional patterns in 5'UTR. 5 SNPs in 3'UTR of the HLA-G gene were predicted to affect the miRNA target sites. Kaplan-Meier analysis showed the HLA-G deregulation can serve as a prognostic marker for some cancers. CONCLUSIONS: The implementation of in silico SNP prioritization methods provides a great framework for the recognition of functional SNPs. The results obtained from the current study would be called laboratory investigations.