Current Pharmaceutical Biotechnology - Volume 23, Issue 11, 2022

Liquid or injectable platelet-rich fibrin (PRF) is a second-generation platelet concentrate that is completely autologous and free of external additives, like bovine thrombin and calcium chloride. Additionally, it is the only one to be obtained in a liquid form among the second-generation platelet concentrates. This allows for wide applications such as maximizing injections or mixing with biomaterials such as bone grafts or antibiotics. Since it was first introduced in 2015, several modifications of the original protocol have been proposed which aim at maximizing its biological and mechanical properties. This includes changes in centrifugation speed, time, and so on. The aim of this review is to summarize the various modifications of the injectable/liquid formation of PRF as well as to discuss the potential applications and future research direction.

Saffron is one of the most expensive spices in the world, and its popularity as a tasty food additive is spreading rapidly through many cultures and cuisines. Minerals and heavy metals are minor components found in saffron, which play a key role in the identification of the geographical origin, quality control, and food traceability, while they also affect human health. The chemical elements in saffron are measured using various analytical methods, such as techniques based on spectrometry or spectroscopy, including atomic emission spectrometry, atomic absorption spectrometry, inductively coupled plasma optical emission spectrometry, and inductively coupled plasma mass spectrometry. The present study aimed to review the published articles about heavy metals and minerals in saffron across the world. To date, 64 chemical elements have been found in different types of saffron, which could be divided into three groups of macro-elements, trace elements, and heavy metals (trace elements with a lower gravity/greater than five times that of water and other inorganic sources). Furthermore, the chemical elements in the saffron samples of different countries have a wide range of concentrations. These differences may be affected by geographical conditions such as physicochemical properties of the soil, weather, and other environmental conditions like saffron cultivation and its genotype.

Cardiovascular diseases, like coronary heart disease or artery disorders (arteriosclerosis, including artery solidification), heart failure (myocardial infarction), arrhythmias, congestive heart condition, stroke, elevated vital signs (hypertension), rheumatic heart disorder, and other circulatory system dysfunctions are the most common causes of death worldwide. Cardiovascular disorders are treated with stenting, coronary bypass surgery grafting, anticoagulants, antiplatelet agents, and other pharmacological and surgical procedures; however, these have limitations due to their adverse effects. Fibrinolytic agents degrade fibrin through enzymatic and biochemical processes. There are various enzymes that are currently used as a treatment for CVDs, like streptokinase, nattokinase, staphylokinase, urokinase, etc. These enzymes are derived from various sources, like bacteria, fungi, algae, marine organisms, plants, snakes, and other organisms. This review deals with the fibrinolytic enzymes, their mechanisms, sources, and their therapeutic potential.

Background: The plant, Cannabis sativa, is heavily explored and researched with many industrial and pharmaceutical applications. The medicinal and therapeutic role of Cannabis sativa has been summarized in the paper, citing its mechanism of action and influence on the human body. Diseases like metabolic disorders, infectious diseases, and psychological disorders pose negative and long-term drastic effects on the body like neurodegeneration and other chronic system failures. Several existing studies have proved its effectiveness against such diseases. Objectives: This review aims to provide an overview of the role of cannabinoids in various diseases like metabolic disorders, infectious diseases, and psychological disorders. Methods: Various e-resources like Pubmed, Science Direct, and Google Scholar were thoroughly searched and read to make an informative, comprehensive manuscript. Here we tried to summarize the therapeutic aspect of Cannabis sativa and its bioactive compound cannabinoids with respect to various diseases. Results: This review highlights the various constituents which are present in Cannabis sativa, the endocannabinoid system, and the role of cannabinoids in various diseases. Conclusion: Recent research on Cannabis has suggested its role in neurodegenerative diseases, inflammation, sleep disorders, pediatric diseases, and their analgesic nature. Therefore, the authors majorly focus on the therapeutic aspect of Cannabis sativa in various diseases. The focus is also on the endocannabinoid system (ECS) and its role in fighting or preventing bacterial, parasitic, fungal, and viral infections.

Female reproductive tract cancers like ovarian, cervical, vaginal, etc. have led to a serious concern for reproductive health as well as an increase in physical and psychological stresses amongst women. Various conventional techniques like surgery, radiation and chemotherapy are employed but possess limitations such as organ toxicity, infection, nausea, vomiting, etc. Also, several nanotechnology-based synthetic vehicle delivery systems like liposomes, nanoparticles, etc. are used but they lack targeting efficiency that results in poor propulsion and control. Therefore, there is a need for naturally-driven drug carriers to overcome such limitations. Sperm-based drug delivery is the new area for targeted delivery that offers self-propulsion to tumor sites, higher biocompatibility, longer lifespan and increased tissue penetration with enhanced localization. Drug-loaded sperm cells are harnessed with micro/nanomotor that will guide them to the intended target site. The critical analysis of the sperm-based drug delivery system was executed and summarized along with the current challenges. This article deals with the art of delivering the anticancer drug to female reproductive cancer sites with proof-of-concept-based research data and critical discussion on challenges in formulating the sperm-based delivery with a future perspective.

Objectives: The Plasmodium parasite is transmitted directly to humans through the Anopheles mosquito bite and causes vector-borne malaria, leading to the transmission of the disease in Southeast Asia, including India. The problem of persistent toxicity, along with the growing incidence of insect resistance, has led to the use of green pesticides to control the spread of the disease in a cost-effective and environment-friendly manner. Based on this objective, this work investigated the larvicidal, pupicidal, and ovicidal activity of Mentha pipertia using a natural nanoemulsion technique. Methods: GC-MS characterized essential oils of Mentha pipertia leaves were formulated as a nanoemulsion for herbal larvicidal, pupicidal, and ovicidal activities. Size of the nanoemulsion was analyzed by photon correlation spectroscopy. The herbal activities against Anopheles Stephensi of nanoemulsion were evaluated in terms of the lethal concentration for 50% (LC50) and 90% (LC90) to prove low cost, pollution free active effective formulation. Results: Chiral, keto, and alcohol groups are obtained from Mentha pipertia leaves’ essential oil, and the nanoemulsions have demonstrated good results in the larvicidal probit analysis, with values of LC50=09.67 ppm and LC90=20.60 ppm. Activity results of the most stable nano formulation with 9.89 nm size showed a significant increase when compared to the bulk. Conclusion: The nanoemulsion of Mentha pipertia leaves can be a promising eco-friendly widely available, low-cost herbicide against the Anopheles mosquito.

Background: Shortage of oxygen is a common condition for residents of high-altitude (HA) areas. In mammals, hemoglobin (Hb) has four derivatives: oxyhemoglobin (Hb-O²), carboxyhemoglobin (Hb-CO), sulfhemoglobin (Hb-S), and methemoglobin (Met-Hb). In HA areas, aberrant physiological performance of blood hemoglobin is well-established. Objectives: The study aimed to investigate the influence of 30 days of HA residence on rabbits' total Hb, Hb derivatives, Hb autooxidation rate, and antioxidant enzymes in comparison to low-altitude control rabbits. Further, the study aimed to investigate the effect of antioxidant-rich Angelica archangelica and/or Ginkgo biloba extracts on the same parameters in HA-resident rabbits. Methods: Rabbits subjected to 30 days of HA residence were compared to low-altitude control rabbits. HA-residence rabbits were then orally administered 0.11 g/kg b.wt. of Angelica archangelica and/or Ginkgo biloba extract for 14 days. Hb derivatives and Hb autooxidation rate were measured spectrophotometrically. Antioxidant enzymes were estimated using specialized kits. Results: Compared to low-altitude rabbits, 30-day HA-residence rabbits showed a noticeable increase (p<0.05) in Hb-O2 and Hb-CO concentration. In addition, Met-Hb concentration, autooxidation rate of Hb molecules, and activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx) exhibited a remarkable increase in HA-residence rabbits (p<0.01), reflective of rapid ROS generation. In HA-residence rabbits, both individual and combined treatment with antioxidant-rich extracts for 14 days resulted in recovery to near-normal functional levels of Hb-O2 and Met-Hb, Hb autooxidation rate, and activities of SOD and GPx, while only combined treatment led to Hb-O2 recovery. Conclusion: The findings suggest that functional Hb levels may be recovered by oral administration of A. archangelica, G. biloba, or combined treatments. In conclusion, oxidative stress due to living in HA areas may be avoided by supplementation with natural antioxidants.

Background: Leonotis nepetifolia (L.) R. Br. (Lamiaceae) is a shrub traditionally used to alleviate inflammatory conditions. Objectives: The present study aimed at investigating the biological activity of methanolic nontransformed and transformed Rhizobium rhizogenes root extracts from L. nepetifolia against human melanoma cells. Methods: Cytotoxicity and genotoxicity properties, the impact on topoisomerase I activity, and proapoptotic activity were evaluated by the MTT test, comet assay, topoisomerase I assay, and fluorescence-activated cell sorting analysis. Moreover, the expressions of p53 were examined by qPCR and Western blot analysis. Docking studies were conducted to assess the potential interactions of the identified phytochemicals with the p53 binding protein Mdm-2, and computational analyses exhibited their antioxidant potential. Results: Both extracts showed cytotoxic potential against human melanoma cells, but generally the activity was more potent for transformed roots than untransformed (IC50 760 μg/mL and 980 μg/mL, respectively). A similar effect was revealed during the evaluation of genotoxic and proapoptotic properties. Moreover, the expression of p53 was also found to be increased after extract treatment. The most dominant identified compounds in both extracts were as follows: (+)- catechin, p-coumaric acid, m-coumaric acid, and (+)-rosmarinic acid. Docking studies and computational analysis showed that (+)-rosmarinic acid possesses the highest binding affinity to the p53 binding protein, Mdm-2, and exhibits the best antioxidant property from the most commonly identified phytochemicals. Conclusion: Our findings revealed the potential of L. nepetifolia transformed root extract as a source of bioactive compounds with cytotoxic, genotoxic, and proapoptotic activity against human melanoma cells as well as antioxidant properties.

Background: Autoimmune thyroiditis (AIT), a T cell-mediated organ-specific disorder, and transcription factors have a critical role in the regulation of immune responses, especially in the fate of T-helper cells. Objectives: This study aims to investigate changes in the gene expression profile of transcription factors and regulators in patients with different forms of thyroid pathology. Methods: We used the pathway-specific real-time PCR array (Neurotrophins and Receptors RT2 Profiler PCR Array, QIAGEN, Germany) to identify and verify transcription factors and regulators pathway-focused genes expression in peripheral white blood cells of patients with postoperative hypothyroidism, hypothyroidism as a result of AIT and AIT with elevated serum and antithyroglobulin (anti-Tg) and anti-thyroid peroxidase (anti-TPO) antibodies. Results: It was shown that in patients with postoperative hypothyroidism FOS, NR1I2, STAT4, and TP53 significantly increased their expression, whereas the expression of STAT1, STAT2, and STAT3 decreased. In patients with hypothyroidism as a result of AIT, we have found increased expression of NR1I2, STAT2, and STAT3. In contrast, the expression of STAT1 and TP53 decreased. FOS and STAT4 mRNAs did not change their expression. In patients with AIT and elevated serum anti-Tg and anti-TPO antibodies, the expression of FOS and NR1I2 reduced, whereas the mRNA level of STAT3 increased. STAT1, STAT2, and STAT4 mRNAs did not change their expression. MYC did not change its expression in all groups of patients. Conclusion: The results of this study demonstrate that autoimmune thyroiditis and hypothyroidism affect the mRNA-level expression of transcription factors and regulators genes in a gene-specific manner and that these changes to genes expression can be one of the triggers of autoimmune inflammation progression in the thyroid gland.

Background: Our study seeks to obtain data to assess the impact of circPUM1 on pancreatic cancer (PC) and its mechanism. Methods: The expression of circPUM1 and miR-200c-3p in PC and normal tissues and PC cell lines was collected and detected, and subsequently dual-luciferase assay-based verification of the binding site of the two was carried out. After interfering with circPUM1 expression in MIAPaCa-2 and PANC-1 cells, cell proliferation, viability, apoptosis rate, invasion ability, glucose consumption, and lactate production were measured by MTT, colony formation, flow cytometry, Transwell assays, and glucose and lactate assay kits. Additionally, western blot was utilized for assessing PI3K/AKT signaling pathway-related proteins. From the results, highly expressed circPUM1 and miR-200c-3p in PC tissues and cells were proved. Results: Down-regulation of circPUM1 expression significantly inhibited cell proliferation, cell viability, invasion and glycolysis, while increasing the apoptosis rate. Down-regulated circPUM1 led to the inhibition of the PI3K/AKT signaling pathway activity in PC cells; while up-regulated circPUM1 increased its activity. Further experiments revealed that down-regulation of miR-200c-3p expression reversed the inhibitory effect of lowly expressed circPUM1 on PC cells. Conclusion: In summary, circPUM1 activates PI3K/AKT signaling pathway by sponging miR-200c-3p and promotes PC progression.