Iranian Biomedical Journal
Volume:26 Issue: 6, Nov 2022

Lung cancer remains a major factor contributing to morbidity and mortality worldwide. Apart from the chemotherapeutic agents in routine use, factors targeting novel molecular pathways are in clinical trials and provide hope for terminal lung cancer patients. The endocannabinoid system has recently become a popular field of study. Many experimental studies have shown that CBD and THC could be useful as a specific treatment for lung cancer since they play a major role in lung cancer cell apoptosis. The objective of this review was to evaluate the antitumorigenic mechanisms of CBD in lung cancer cells.

We searched the databases MEDLINE, clinicaltrials.gov, Cochrane Central Register of Controlled Trials, and Google Scholar using specific terms. A total of 246 studies were screened, and nine studies were included in the review. All the selected studies were conducted in vitro, and four of which also had an in vivo component. Included studies were assessed in our review using the ToxRTool.

The most common cell line used in all of the studies was A549; however, some studies contained other cell lines, including H460 and H358. We concluded that CBD has direct antineoplastic effects on lung cancer cells through various mechanisms mediated by cannabinoid receptors or independent of these receptors. All studies were referred to an in vitro model; hence, further research in animals is required, and if results remain promising, human clinical trials are ncessary.

Downstream processing of therapeutic recombinant proteins expressed as the IBs in E. coli is quite challenging. This study aimed to use the QbD approach for developing the multi-step downstream process of a structurally complex therapeutic Fc-Peptide fusion protein, romiplostim.

For the development of a successful downstream process, risk analysis, and experimental designs are used to characterize the most CQAs and effects of process parameters on them.

The solubilization of IBs was optimized by DoE on three parameters with a focus on solubility yield, and >75% of the target protein was solubilized. The pH of sampie was identified as (CQA) inanion exchange charmatography that might have an impact on achieving > 85% host cell proteins removal and >90% host cell DNAs reduction. In refolding step, process parameters were screened. Cystine/cysteine ratio, pH, and incubation time Were identified as CPPs were further optimized using Box-Behenken analysis, and > 85% of target protein was refolded. The design space for futher purification step by HIC was mapped with a focus on high molecular weight impurities. after polishing by gel filtration, the final products biological activity had no statistically significant differences among the groups received romiplostim and Nolate as the reference product. 

This research presents a precise and exhaustive model for mapping the design space in order to describe and anticipate the link between the yield and quality of romiplostim and its downstream process parameters.

Ferritin has an important role in iron storage in the cells, and due to its nanocage structure and self-assembly properties, it has wide application prospects in nanobiotechnology.

The maize (Zea mays) ferritin gene ZmFer1 was cloned and expressed in Escherichia coli BL21 (DE3) for the first time. Change in macromolecular structure of ZmFer1 ferritin due to heat treatment was investigated using native PAGE electrophoresis, DLS, and TEM. Change in the secondary structures of the protein was evaluated using CD spectroscopy. Moreover, alteration in the conformation of the protein was evaluated using UV-absorption spectra and intrinsic fluorescence spectra. The Tm of ZmFer1 was obtained using DSC. Finally, the effect of heat on the function of ZmFer1 was assessed by iron loading ability.

The purified ZmFer1 protein showed a homopolymer nanocage structure. The results of native PAGE electrophoresis, DLS, and TEM techniques showed that ZmFer1 protein nanocage is stable to heat treatment up to 90 °C, and some of the protein nanocages retain their macromolecular structures even at 100 °C in liquid aqueous solution. Based on the DSC results, ZmFer1 protein nanocage had a Tm of 81.9 °C. After treatment at 100 °C, stable ZmFer1 protein nanocages were able to store iron atoms.

Recombinant ZmFer1 ferritin with a Tm > 80°C is a hyperthermostable protein nanocage. The results of this study are beneficial for the  development of protein nanocages that are stable under extreme temperature conditions, as well as application of ZmFer1 in nanobiotechnology, biomaterials, and biomedical fields.

INPP5A is involved in different cellular events, including cell proliferation. Since INPP5A, HLAG1, IL-10, and MMP-21 genes play fundamental roles in ESCC tumorigenesis, we aimed in this study to clarify the possible interplay of these genes and explore the potential of these chemistries as a predictor marker for diagnosis in ESCC disease.

Gene expression analysis of INPP5A, HLAG-1, IL-10, and MMP-21 was performed using relative comparative real-time PCR in 56 ESCCs compared to their margin normal tissues. Immunohistochemical staining was accomplished for INPP5A in ESCCs. Analysis of ROC curves and the AUC were applied to evaluate the diagnostic capability of the candidate genes.

High levels of HLA-G1, MMP-21, and IL-10 were detected in nearly 23.2%, 62.5%, and 53.5% of ESCCs compared to the normal tissues, respectively, whereas INPP5A underexpression was detected in 19.6% of ESCCs, which all tested genes indicated significant correlations with each other. The protein expression level of INPP5A in ESCC tissues was significantly lower than that of the non-tumor esophageal tissues (p = 0.001). Interestingly, the concomitant expression of the INPP5A/HLA-G1, INPP5A/MMP-21, INPP5A/IL-10, HLA-G1/MMP-21, HLA-G1/IL-10, and MMP-21/IL-10 was significantly correlated with several clinicopathological variables. INPP5A, HLA-G1, MMP-21, and IL-10 showed to be the most appropriate candidates to discriminate tumor/non-tumor groups due to the total AUCs of all combinations (>60%).

Our results represent a new regulatory axis containing INPP5A/HLAG-1/IL-10/MMP-21 markers in ESCC development and may provide novel insight into the mechanism of immune evasion mediated
by the INPP5A/HLAG-1/IL-10/MMP-21 regulatory network in the disease.

Hyaluronic acid (HA), a natural polymer with unique properties and wide application, is mainly produced by Streptococcal fermentation at industrial scale. In the present work, two rounds of chemical random mutagenesis were applied on Streptococcus equisimilis group G to obtain high-producing strains using 1-methyl-3-nitro-1-nitrosoguanidine. The non-hemolytic and hyaluronidase-negative mutants were screened on blood agar and HA agar, respectively. HA productivity and molecular weight (Mw) was determined by carbazole assay and agarose gel electrophoresis, respectively. Moreover, the stability of strains was assessed within 10 generations in term of HA productivity. The findings showed that the wild type strain produced 1241 ± 2.1µg/ml HA at pH 5.5 and 4 hours of cultivation, while the screened mutants showed a 16.1 - 45.5 % increase in the production. Two mutant strains, named Gm2-120-21-3 (2470± 8.1µg/ml) and Gm2-120-21-4 (2856± 4.2µg/ml), showed the highest titer and a consistent production. The molecular weight of HA for the mutants was less than 160 kDa which considered a low Mw. The mutant strains producing a low polydisperse and low Mw HA at high titer can be considered potential industrial strains for HA production after further safety investigations.

Bone tissue engineering has shown a promising way in order to renew bone defects without conflicting with previously known side effects. Three main building blocks including seeding cells, scaffold, and signaling molecules are required for this process. The human amniotic membrane (hAM) is the innermost of the placental membranes. As well as providing a source of stem cells and growth factors, hAM has several features which make it an appropriate stem cell containing scaffold in order to be used in orthopedic surgery. The present investigation aimed to demonstrate the effect of bone morphogenetic protein-9 (BMP-9) alongside phenamil and simvastatin on osteogenic induction of hAM with its sessile epithelial cells (hAECs). For this purpose, hAM was cultured using six different osteogenic media for 14 days. The basic osteogenic media was chosen as the first group and other media were made by addition of BMP-9, phenamil, simvastatin, BMP-9 alongside phenamil, and BMP-9 alongside simvastatin to the basic osteogenic media. Finally, viability, mineralization, calcium and phosphate content, lactate dehydrogenase (LDH), and alkaline phosphatase (ALP) activity were evaluated. Among all study groups, simvastatin-containing groups showed a significantly lower level of viability. Although, all media could induce osteogenic features, the hAECs which were cultured in media containing BMP-9 and phenamil could demonstrate a wider area of mineralization and a significantly higher level of calcium, phosphate, LDH, and ALP activity. These findings indicated that the application of phenamil alongside BMP-9 could synergistically show in situ osteogenic induction in hAECs.

Chemotherapy, biotherapy, and radiotherapy play a limited but important role in treating breast cancer. For more efficient treatment, combination therapy could be an appropriate option. In this study, radiotherapy using neutron radiation emitted from a 241Am-Be neutron source, as well as biotherapy using curcumin (80 μM) was combined to investigate the efficiency of treatment towards MCF-7 breast cancer in a 3D culture medium.

MTT, NR uptake assay, NO, GSH assay, catalase, cytochrome c, comet assay, and caspase-3 were used to determine the effect of neutron radiation and also neutron and curcumin combination on the viability of cancer cells.

The results of cytotoxicity test showed that neutron irradiation with or without curcumin at 5, 10, 15, and 20 h reduced the survival of tumor cells. Moreover, the rate of apoptosis due to the neutron effect at different irradiation times enhanced with the increasing time.

Due to the significant anticancer effect of curcumin in 3D culture, using this molecule before or after neutron therapy is recommended.

In spite of many reports on persistent low CD4 T cell counts and change in immune-related gene expression level in patients with HIV infection, there is still uncertainty about significant association between gene expression level and HIV infection in patients with and without DIR. The aim of this study was to compare the expression level of CD4, CCL5, IFN-γ, STAT1, APOBEC3G, CD45, and ICAM-1 genes in HIV-1-positive patients with and without DIR.

In this study, 30 HIV-1-positive patients (15 patients with and 15 patients without DIR [control group]) were included. PBMCs of the patients were collected through density radient centrifugation with Ficoll-Hypaque. RNeasy Plus Mini kit was used to extract RNA. Relative expression levels of CD4, CCL5, IFN-γ, STAT1, APOBEC3G, CD45, and ICAM-1 genes were evaluated by real-time PCR.  The data were analyzed using one-way ANOVA.

CD4 T cell counts were significantly lower in DIR patients than the control group (p < 0.01). While there was no significant difference in the relative expression levels of CD4, CCL5, IFN-γ, STAT1, CD45, and ICAM-1 between patients with DIR and control group, APOBEC3G expression level was significantly higher in the patients with DIR as compare to the control group (p < 0.01).

Our findings suggest a significantly higher APOBEC3G expression level in patients with DIR, suggesting the potential role of APOBEC3G in patients with immunological discordance besides its suppressing role in HIV-1 infection. Confirmation of this hypothesis requires further research.