Cellular and Molecular Biology

Assessment of immunological factors in COVID-19 patients treated by convalescent plasma

Mozhdeh Heidari — 2024-10-08

Following the outbreak of COVID-19, several immunotherapy methods were used to modulate the immune responses of patients. In this study, we aimed to evaluate the immune response to COVID-19 in patients receiving convalescent plasma. In this regard, this randomized controlled trial included 30 patients who were divided into two groups according to receiving convalescent plasma or normal control plasma. Samples from both groups were collected on days 0, 1, 3, 5 and 7 after plasma infusion. We measured the expression level of TLR7/8, IRF3/7, CTLA-4, PD-1 and T cell transcription factors by Real-time PCR in the mentioned groups. Thirteen cytokines were also evaluated using flow cytometry method. Results showed that compared to the normal control plasma group, the expression levels of TLR7, 8, IRF3, 7 and PD-1 and CTLA-4, on days 3, 5 and 7 after convalescent plasma infusion, were significantly decreased. On the other hand, Gene expression results showed that the expression levels of Tbet, RORγ3 and Foxp3 on days 3, 5 and 7 after convalescent plasma infusion were significantly increased compared to the normal control plasma group. After convalescent plasma infusion, the viral load was significantly decreased compared to the normal control plasma group. Convalescent plasma infusion also reduced the plasma cytokines levels, including IL-6, IL-10, and IL-4, and enhanced the level of IL-2, IFN- γ and perforin comparing the normal control plasma group. According to the results, the convalescent plasma infusion led to a decrease in the expression of innate immunity receptors and an increase in the expression of transcription factors of adaptive immunity. Therefore, it may be concluded that convalescent plasma infusion can modulate the immune response. To achieve a reliable consequence, further studies are required.

Functional observational battery (FOB) tests using caffeine and chlorpromazine hydrochloride in sprague-dawley rats

Subramanian Baskaran — 2024-10-08

Caffeine is believed to exert its therapeutic effects by acting as a nonselective, competitive antagonist of adenosine receptors. Chlorpromazine, a phenothiazine, is a classic psychotropic mediator extensively used in the clinical administration of psychotic disorders. This study aimed to validate the procedures used for performing Functional Observational Battery (FOB) tests, to demonstrate the proficiency and interobserver reliability during the FOB tests and also to assess effect on neurobehavioral parameters using positive controls in rats. The rats were administered with Caffeine in Milli-Q water as oral gavage at the dose of 20 mg/kg and Chlorpromazine HCl in 0.9% Saline as intraperitoneal route at the dose of 20 mg/kg. No inter-personnel variability was observed in home cage, handling, open field and sensory reactivity observations recorded in Proficiency test. In conclusion, the known effects of positive controls; caffeine and chlorpromazine HCl on neurobehavioral/Functional Observational Battery parameters including autonomic, neuromuscular and sensory reactivity tests were detected in the current study. FOB test procedures for neurobehavioral, grip strength and motor activity are adequate for the detection of neurotoxic effects of positive controls. No major inter-personnel variability was observed between study personnel in neurobehavioural observations.

Anti-inflammatory effects of phytosphingosine-regulated cytokines and NF-kB and MAPK mechanism

Mikyung Sung — 2024-10-08

Phytosphingosine (PHS) is a major component of the skin barrier and a multifunctional physiologically active substance. This study aimed to investigate the types of cytokines regulated by PHS, their anti-skin inflammatory effects, and their anti-inflammatory mechanisms. RAW264.7 cells stimulated with Lipopolysaccharides (LPS) were treated with PHS to measure inflammatory factors such as nitric oxide (NO) and prostaglandin E2 (PGE2), and gene expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX2) were confirmed by q-PCR. Cytokines regulated by PHS against LPS-induced inflammation were found through cytokine array, and each factor was reconfirmed through ELISA. Western blot was performed to confirm anti-inflammatory mechanism of Iκbα and MAPK. To confirm anti-skin inflammatory efficacy, HaCaT cells stimulated with TNF-α/IFN-γ were treated with PHS, and TARC, IL-6, and IL-8 were detected by ELISA. PHS suppressed the gene expression of iNOS and COX2, which were increased by LPS, and suppressed NO and PGE2 production. Through cytokine array, it was confirmed that IL-6, IL-10, IL-27 p28/IL-30, IP-10, I-TAC, MCP-5, and TIMP-1 increased by LPS were decreased by PHS. PHS inhibited NF-κB signaling by inhibiting LPS-induced NF-κB nuclear migration and p-Iκbα-mediated Iκbα degradation, and inhibited p38, ERK, and JNK signaling pathways. PHS reduced the production of TARC, IL-6, and IL-8 increased by TNF-α/IFN-γ. These results indicate PHS has anti-inflammatory effects via the suppression of inflammatory factors and pro-inflammatory cytokines through the NF-κB and MAPK pathways. Moreover, these results may explain beneficial effects of PHS in the treatment of skin inflammatory conditions induced by TNF-α/IFN-γ.

Pro-inflammatory cytokines gene expression in liver and kidneys of rats exposed to a sub-lethal dose of Bitis arietans snake venom

Haseeb A. Khan — 2024-10-08

Bitis arietans (Puff adder) is a poisonous snake and its bite causes pain, edema, blistering, tissue damage and neutrophilia. There are limited studies on inflammatory process involved in Bitis arietans envenomation. We therefore investigated the role of proinflammatory cytokines in Bitis arietans venom (BAV)-induced liver and kidney toxicities in rats. Adult male Sprague Dawley rats were treated with BAV (0.5 mg/kg) and were sacrificed after specific time intervals (2 h, 24 h, 1 week). Blood samples were collected for liver and renal function tests and tissues were collected for histopathology and gene expression analysis of IL-1β, IL-6, and TNF-α in liver and kidneys. There was no significant difference in serum ALT activities among different treatment groups. Serum AST was significantly increased at 24 h following BAV injection. In both organs, injection of BAV resulted in mild inflammatory cell infiltration at 2 h post-dosing which normalized after 1 week. In liver, there was a significant increase in IL-1β expression in BAV-treated rats at 2 and 24 h post-dosing that reduced after one week. Significant increases in IL-6 and TNF-α were observed at 24 h and 1 week after BAV exposure. In kidneys, there were significant increases in IL-1β and TNF-α expression at 24 h that subsided after 1 week. In conclusion, a single sub-lethal dose of BAV caused an acute phase inflammation in liver and kidneys. It is most probable that a higher dose of BAV may result in greater and irreversible damage to these organs.

mTORC2 inhibition by JR-AB2-011 improves IL-1β-induced inflammation, catabolic response, and apoptosis in human chondrocytes through IκB-α/NF-κB p65

Meryem Temiz-Resitoglu — 2024-10-08

Osteoarthritis (OA) is a very common chronic joint condition marked by inflammation and cartilage loss. mTOR is a well-known mediator of inflammation, cell survival, and aging; however, its role in OA has not been determined. To explore the role of mTORC2 in OA-and associated pathological changes, we examined the contribution of mTORC2-mediated Akt, rictor and IκB-α/NF-κB p65 pathway in interleukin (IL)-1β-treated human chondrocytes. We focused on the protein expression of proinflammatory cytokines and catabolic and apoptotic factors, including TNF-α, IL-6, iNOS, MMP13, Bax, and caspase3, which may occur through this signalling pathway in IL-1β-treated chondrocytes. Chondrocytes were cultured and treated with either 2 ng/mL IL‑1β alone or in combination with increasing concentrations of JR-AB2-011 (50, 100, or 250 µM), a selective mTORC2 inhibitor. The protein levels of phosphorylated (p)‑Akt, Akt, rictor, p-NF-κB p65, NF-κB p65, IκB-α, p-IκB-α, iNOS, MMP13, Bax, and caspase3 were evaluated by Western blotting. In IL-1β-stimulated chondrocytes, mTORC2 activity was increased with increased phosphorylation of Akt and expression of rictor. IL-1β increased the expression of p-IκBα, p-NF-κB p65, NF-κB p65, IL-6, TNF-α, iNOS, Bax, and caspase3 proteins and decreased the expression of IκB-α. All of these IL-1β-induced alterations were prevented by JR-AB2-011. The main novel finding in the present study is that selective mTORC2 inhibition by JR-AB2-011 prevents the inflammatory, catabolic, and apoptotic responses induced by IL-1β via modulation of IκB-α/NF-κB activity. Therefore, we demonstrated a previously unknown function of mTORC2 inhibition that seems to be a potential therapeutic target for OA.

Histological assessment of potential inferior alveolar nerve injury following osteotomy of the mandibular buccal cortex using a piezoelectric saw

Shamal Hassan Qadir — 2024-10-08

The present study aimed to evaluate the effect of Piezosurgery on histopathologic features of the inferior alveolar nerve (IAN) damage after osteotomy of the buccal cortex of the mandible using piezoelectric devices in Hamdani sheep. A total of ten healthy mature female sheep were included. Each side of the mandible underwent two different experiments: the first experiment operated directly on the mental nerve by touching and activating the piezo tip on the nerve for ten seconds for the left side and thirty seconds for the right side. In the second experiment, the inferior alveolar nerve was touched by an activated piezo tip inside the mandibular canal for ten seconds on the left side and thirty seconds on the right side. All the nerve samples underwent histopathological evaluation, and the scoring system was performed to assess the nerve structures. Mental nerves exposed to piezo tip for 10 seconds showed mild abnormality including disruption of the perineurium with the endoneurium remaining intact. Mental nerves exposed for 30 seconds showed moderate injury with destruction of the perineurium and moderate degeneration of nerve fibers, nevertheless, the endoneurium remained continuous with normal node of Ranvier. Severe damage of the inferior alveolar nerve was seen after exposure to piezo tip for 10 seconds, which showed sloughing of the perineurium and severe vacuolar degeneration of nerve fibers, partial disruption of the endoneurium; however, the axons were still intact. Inferior alveolar nerves exposed for 30 seconds revealed destruction of the perineurium, marked vacuolar degeneration of nerve fibers, focal damage of axon and loss of endoneurium (axonotmesis). Piezosurgery devices have the potential to cause severe nerve damage during surgery and should be used very carefully.

Seasonal distribution and correlation between IL-10 and IL-13 gene polymorphism and their expression in scabies-infected patients

Eman Farhad Bilal — 2024-10-08

Scabies is a significant concern in global health. It is more prevalent in individuals who have poor hygiene and live in crowded conditions, hence, it is seasonal distribution and immunological response in Erbil population is the aim of the present study. In the Erbil Dermatology Education Centre in Erbil, Iraq, 154 patients were recruited for the research between April 2022 and March 2023. If a patient has a suspicious skin lesion and itching for a minimum of one week, scabies may be considered. Blood samples were collected from each participant in the study to evaluate serum levels of IL-10 and IL-13, then the DNA was isolated to study the gene polymorphism for the mentioned cytokines. Results showed that female 60.3% were more infected than male 39.6%. The median age of participants was (10 - >51) years, among infested, adolescents aged 10-20 years displayed the highest rate (31.8%). The carriers of GA genotype of IL-10 were protective against the infection, OR:0.61. while the TT carriers of IL-13 were susceptible to scabies infection with OR:2.14. IL-10 GA genotype was more prevalent in male patients OR:2.14 whereas the AA genotype was most protective in females OR:0.32. the IL-13 CT genotype was protective for males with OR:0.52. Both of IL-10 and IL-13 serum levels were increased significantly with infection and highest levels were found in wild homozygous genotypes (GG and CC) and lowest ratio was found in mutant homozygous genotypes of IL-10 and IL-13 respectively. Point mutation in IL-10 GA was protective and wild TT genotype of IL-13 was susceptible to the scabies infection. Double mutation in IL-10 AA was protective to females and single mutation of IL-13 CT was protective to males.

Generation of dengue 3 envelope domain III using tobacco mosaic virus-based vector system and its immunological response mouse model by generating anti-dengue virus antibodies

Hailah M. Almohaimeed — 2024-10-08

Producing recombinant proteins in plants has become a valuable alternative to traditional microbial or mammalian systems due to its cost-effectiveness, scalability, and ability to perform post-translational modifications. This study investigates the use of the Tobacco Mosaic Virus (TMV)-based vector system for producing the Dengue virus serotype 3 (DENV-3) envelope domain III (EDIII) protein in plants.. A fragment of the gene that encodes domain III of the dengue 3 envelope protein (D3EIII, comprising 300-420 amino acids), was effectively expressed within Nicotiana tabacum plants utilizing a transient expression system based on tobacco mosaic virus (TMV). The N-terminal 5' UTR region upstream of D3EIII notably enhanced protein yield in infected tissues. The produced recombinant protein exhibited reactivity with both (anti) D3EIII polyclonal antibodies and antibodies of anti-His tag. Upon injection of EDIII in mice, it stimulated the generation of antibodies against the dengue-specific virus. The induced antibodies demonstrated neutralizing activity against dengue virus type 3. These findings indicate that the TMV expression system is effective for producing dengue virus antigens in plants, resulting in antigens with appropriate properties and strong immunogenic potential.

CRISPR-Cas9 guided RNA-based model for the silencing of spinal bulbar muscular atrophy: A functional genetic disorder

Muhammad Naveed — 2024-10-08

This study explores a novel therapeutic approach for spinal bulbar muscular atrophy (SBMA), a neurodegenerative disorder caused by a mutation in the Androgen Receptor (AR) gene. The aim is to investigate the potential of CRISPR-Cas9 technology in targeting the mutant AR gene to inhibit its production. The objectives include assessing the accuracy and efficacy of CRISPR-Cas9 guided RNAs in silencing the mutant gene and evaluating the feasibility of this approach as a treatment for SBMA. Computational and in-silico approaches are used to evaluate the feasibility of using CRISPR-Cas9 technology for treating SBMA. Computational analysis is used to design CRISPR-Cas9 guided RNAs targeting the mutant AR gene, assessing their on-target and off-target scores, GC content, and structural accuracy. In-silico simulations predict the potential therapeutic outcomes of the CRISPR-Cas9 approach in an artificial environment. Three guided RNA (gRNA) sequences were designed using the CHOPCHOP tool, targeting specific regions of the AR gene with high efficiency and 100% match. These gRNAs demonstrated effective targeting with minimal off-target scores and optimal GC content. Additionally, lentiCRISPR v2 plasmids were designed for the delivery of CRISPR materials, enabling high-efficiency multiplex genome editing of the AR gene. Thermodynamic ensemble predictions indicated favorable secondary structure stability of the designed gRNAs, further supporting their suitability for gene editing. The evaluation of designed gRNAs confirmed their strong binding ability to the target sequences, validating their potential as effective tools for genome editing. The study highlights the potential of CRISPR-Cas9 technology for targeting the Androgen Receptor gene associated with spinal bulbar muscular atrophy (SBMA). The findings support the feasibility of this approach for gene editing and suggest further exploration in preclinical and clinical settings. Recommendations include continued research to optimize CRISPR-Cas9 delivery methods and enhance specificity for therapeutic applications in SBMA.

Effects of using different dentin conditioners on dentin regeneration

Hiwa Saeed Khidir — 2024-10-08

This experiment aimed to evaluate the impact of several dentine etching and conditioning agents on growth factors (GFs) liberation from dentine slices. Eighteen dentine slices were obtained from nine premolars divided in to six groups, the slices immersed in one mL test solutions for 5 min; Group 1: white Mineral trioxide aggregate (MTA), Group 2: Phosphate buffered saline (PBS), Group 3: 37% phosphoric acid, Group 4: 17% Ethylenediaminetetraacetic Acid (EDTA), Group 5: 10% Maleic acid (MAc), and Group 6: 0.7% Fumaric acid. The solutions were removed and stored directly at for further detection and quantification of transforming GF beta 1 (TGF-b1), bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor (VEGF) by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA was used to compare the mean release and standard deviation between groups (α = 0.05). Tukey’s post hoc applied for multiple comparisons. After five min conditioning of dentine slices, white MTA released the highest level of TGF-b1, BMP2 and VEGF among all groups, followed by 0.7% Fumaric acid with no significant difference between them, but compared to 37% phosphoric acid and PBS groups significant difference observed, which they released the least amount of GFs amongst all groups. Based on the results of this research the detectable release of TGF-b1, BMP2 and VEGF by 0.7% fumaric acid was comparable with white MTA from dentin slices.

Morphological and molecular identification of freshwater eutardigrade Dactylobiotus parthenogeneticus (Bertolani, 1982) in the Greater Zab River of Kurdistan Region – Iraq

Khasro Abdulrahman Ismael — 2024-10-08

Dactylobiotus parthenogeneticus is one of the widespread species of tardigrade all over the world. Tardigrades of this species were collected from the Greater Zab River in Erbil City-Iraq by filtering water of the river through a plankton net with a mesh of 45 µm pore. The samples were mounted on a slide with a cover slip and examined under the microscope to determine morphological characteristics and measurements. Based on these characters the species identified to be D. parthenogeneticus. To support this diagnosis, DNA barcoding techniques were applied to do molecular analysis and sequencing on the cytochrome oxidase subunit I (COI) gene. The sequence was subjected to the GenBank database of NCBI and recorded with the accession number PP140905. The result of the sequencing and molecular analysis of the cytochrome oxidase subunit I (COI) gene confirmed to be the same species diagnosed by relying upon morphological characters. This study represents one of the pioneer researches and documents on tardigrades and found D. parthenogeneticus for the first time in the Greater Zab River in Kurdistan, North of Iraq. Tardigrades play a magnificent role in different trophic levels and can be utilized as an indicator of ecosystem health.

Identification of an immune cell infiltration-related gene signature for prognosis prediction in triple-negative breast cancer

Yan Wang — 2024-10-08

Triple-negative breast cancer TNBC with higher immunogenicity and tumor-infiltrating lymphocyte (TIL) enrichment can benefit from immunotherapy relative to other breast cancer subtypes. Our work was designed to identify the TIL-related hub genes in TNBC and construct a prognostic signature for TNBC. TNBC gene expression files were obtained from the TCGA database. CIBERSORT algorithm and random forest risk model were used for immune infiltration group division. The TIL-related differentially expressed genes (DEGs) were then selected and subject to GO, KEGG analyses and GSEA. Next, Lasso cox regression analyses were adopted for constructing a prognostic risk model, followed by evaluation using time-dependent ROC curves. The copy number variation between the two risk groups was also analyzed, and major genomic mutation types were identified. Additionally, the nomogram was constructed with calibration curve for clinical prognosis analysis. Our results showed that totally 113 TNBC samples were allocated into the high or low-immune risk groups. We identified 243 DEGs between groups, namely TIL-related DEGs, with 128 upregulated and 115 downregulated genes. Among the TIL-related DEGs, 6 hub genes (SLITRK3, PCDHGB3, NELL2, SRRM4, ASIC2 and B4GALNT2) were screened out and constructed a prognostic risk signature, which had good performance for long-term prognosis prediction. Analysis of genomic mutation showed that the TP53, PIK3CA, TTH, etc. showed high mutation frequency in the two prognostic risk groups. Moreover, the higher risk score of the prognostic risk model predicted poor overall survival in TNBC patients, and nomogram and calibration curve confirmed the potent prediction ability of this model. To sum up, six TIL-related biomarkers (SLITRK3, PCDHGB3, NELL2, SRRM4, ASIC2 and B4GALNT2) were identified and used for the construction of the prognostic risk model, which might provide novel insight for the clinical decisions.

LncRNA PCBP1-AS1 suppresses cell growth in oral squamous cell carcinoma by targeting miR-34c-5p/ZFP36 axis

Orkideh Shafiee allaf — 2024-10-08

Oral squamous cell carcinoma (OSCC) is the most frequently diagnosed oral malignancy and poses a great threat to public health. According to bioinformatics analysis, long noncoding RNA PCBP1-AS1 is downregulated in OSCC. In this work, the functions and mechanism of PCBP1-AS1 in OSCC were further investigated. PCBP1-AS1 expression in OSCC cells was measured by quantitative polymerase chain reaction. Cell viability and proliferation were detected using CCK-8 assays and colony-forming assays. TUNEL assays as well as flow cytometry analyses were carried out to detect OSCC cell apoptosis. Binding relationship between PCBP1-AS1 and miR-34c-5p or that between miR-34c-5p and ZFP36 in OSCC cells was identified using RNA immunoprecipitation assays, RNA pulldown assays, and luciferase reporter assays. Experimental results revealed that PCBP1-AS1 was downregulated in OSCC cells. PCBP1-AS1 overexpression hampered cell proliferation and enhanced cell apoptosis in OSCC. PCBP1-AS1 interacted with miR-34c-5p in OSCC and negatively regulated miR-34c-5p. ZFP36 3’untranslated region was targeted by miR-34c-5p. PCBP1-AS1 positively regulated ZFP36 expression. ZFP36 silencing abrogated the suppressive impact of PCBP1-AS1 on OSCC cell growth. In summary, PCBP1-AS1 suppresses cell growth in OSCC by upregulating ZFP36 through interaction with miR-34c-5p.

Antibacterial, phytochemical and GC-MS analyses of argel (Solanum argel) leaves

Abdelmuhsin Abdelgadir Abdelmuhsin — 2024-10-08

Finding novel, efficient antimicrobial drugs is crucial in this age of pressing global health challenges. The medicinal qualities of the leaves of the argel plant (Solanum argel, or S. argel) have been recognized in traditional medicine for quite some time. The medicinal potential of these leaves may be due to the presence of bioactive substances such as alkaloids, flavonoids, and phenolic acids. S. argel leaf antibacterial, phytochemical, and gas chromatography–mass spectrometry (GC–MS) characteristics are the focus of this investigation. To conduct the study, bioactive compounds would be extracted from the leaves and tested against a panel of bacterial pathogens. Then, the compounds would be identified using GC-MS analysis. Mean inhibition zones of 15.30±1.0 mm, 14.67±0.42 mm, 15.0±0.01 mm, and 15.56±0.22 mm for the bacteria E. coli, Staph. aureus, and Sal. typhimurium, respectively, were seen in the antibacterial results at a concentration of 3 µg/disc. Secondary metabolites such as alkaloids, flavonoids, phenolic substances, and tannins were identified using phytochemical investigation. Antimicrobial, antioxidant, and anti-inflammatory are just a few of the many bioactivities associated with these phytochemicals. Argel plant leaves contain bioactive chemicals that show they could be a source of new pharmaceuticals. Argel leaves were analyzed using GC-MS and 37 different chemicals were found. The most abundant compounds were 4H-Pyran-4-one and 2,3-dihydro-3.5-hydroxy, followed by 3-Pentanol, 2,2,4,4-tetramethyl, and 2,2-Dimethyl-3-[3-methyl-5-(phenylthio)-, with areas of 11.80%, 10.6%, and 9.47%, respectively. The analysis was performed within a time range of 5.070 to 34.464 minutes. According to the research, Argel leaf has powerful antioxidant and antibacterial capabilities, making it an excellent substance for medical and food preservation applications.

Anti-oxidant and antibacterial activities of novel N-sulfonylphtalimide in an ovalbumin-induced inflammation in Wistar rats

Ilhem Silini — 2024-10-08

Phthalimide and N-substituted phthalimide have a special structure that helps them to be pharmaceutically useful and biologically active. In this study, we investigated the antioxidant, anti-inflammatory and antibacterial effects of a synthetic phthalimide-containing derivative in an experimental asthma model. In vitro determination of antioxidant and chelating activity was carried out by spectrophotometric methods. The in vivo antioxidant activity was carried out in Wistar rats sensitized to ovalbumin in the experimental model of asthma. Our results reveal that that the synthesized N-sulfonylphthalimide molecule has a scavenging capacity against the free radical 2,2-diphenyl-1-picryl-hydrazyl (DPPH•) and a chelating activity on ferrous ions and revealed its protective capacity against altered markers of oxidative stress in the experimental asthma model. All the previous results were confirmed by the result of the histopathological study of the liver. Moreover, neo-synthesized N-sulfonylphthalimide 2 showed antibacterial activity against Gram-positive and Gram-negative bacteria with interesting MIC values. Finally, our study highlights the anti-inflammatory, anti-asthmatic and antibacterial effects of the N-sulfonylphthalimide molecule, which could potentially be a drug of choice in asthmatic pathology, especially during bacterial superinfections in the respiratory tract.

GAS5 promotes glucose metabolism reprogramming and resistance to ferroptosis of endothelial progenitor cells through the miR-495-3p/SIX1 and IGF2BP2/NRF2 dual-regulatory pathways in coronary heart disease

Ming Zhong — 2024-10-08

We aimed to explore the potential along with mechanism of lncRNA growth arrest-specific 5 (GAS5) in modulating glucose metabolism and ferroptosis of endothelial progenitor cells (EPCs) in coronary heart disease (CHD). CCK-8, flow cytometry, EdU, colony formation, scratch test as well as transwell assays were implemented to assess cell biological behaviors. Glucose uptake testing, lactic acid production assay, and detection of extracellular acidification rate (EACR) together with oxygen consumption rate (OCR) were used to assess glucose metabolism. Iron, GSH and MDA detection were used to measure ferroptosis. Besides, a series of mechanical experiments were implemented to clarify the modulatory relationship between GAS5 and nuclear factor erythroid 2-related factor 2 (NRF2) as well as sine oculis homeobox 1 (SIX1). We found that GAS5 was down-regulated in CHD patients relative to healthy controls. GAS5 depletion repressed EPCs proliferation, migration along with invasion while elevated cell apoptosis. GAS5 promoted the reprogramming of glucose metabolism and inhibited ferroptosis in EPCs. GAS5 affected glycometabolic reprogramming and ferroptosis resistance through regulating SIX1 and NRF2. On the one hand, GAS5 promoted NRF2 mRNA stability through IGF2BP2. On the other hand, GAS5 regulated the miR-495-3p/SIX1 axis in EPCs. To sum up, GAS5 promotes glucose metabolism reprogramming and resistance to ferroptosis of EPCs through the miR-495-3p/SIX1 and IGF2BP2/NRF2 dual-regulatory pathways in CHD.

Early detection of cervical tumor cell of origin through Oncogene-induced senescent HPV-positive cells

Rafal Abdulrazaq Al-Rawi — 2024-10-08

Human Papilloma Virus (HPV) is an oncogenic virus and is the most common cause of cervical cancer. HPV has been shown to induce senescence. Cellular senescence is involved in cancer progression and tumorigenesis. Identification and isolation of cells of tumor origin before tumorigeneses is an important step in cancer prevention and treatment. This study aimed to investigate the early cervical atypical senescent cytological preneoplastic change in non-menopausal women. Cervical smears of 121 patients were randomly selected and included in the study which cytopathologically diagnosed as atypical squamous cells of undetermined significance (AS-CUS) in correlation to HPV status, parakeratosis (PK), p16 immunostaining, enlarged Squamous cells nuclei (ES) and inflammatory cells infiltration (ICI). Results revealed that out of the total 121 patients, 32 cases (26%) were positive for high-risk HPV (HR-HPV), 26 cases (22%) were positive for low-risk HPV (LR-HPV) and 63 (52%) were negative for HPV. HPV infections were significantly associated with age groups (p<0.026), PK (p = 0.043), p16 (p = 0.001), ES (p = 0.002) and ICI (p = 0.049). The positive immunostaining expression of p16 was only noticed in two HR-HPV patients. ES cells were found in 9.5% of HPV-negative cases, 27% of LR-HPV cases and 40.5% of HR-HPV cases. High PK cell positivity was seen only in HR-HPV. High ICI scores were seen in 40.6% of HR-HPV patients, 26.9 % of LR-HPV and 17.4 % of negative HPV patients. It was concluded that high PK positivity, high ICI score, positive p16 immunostaining and ES were correlated with HR-HPV in non-menopausal women. These findings could provide potential diagnostic clues for HPV-harboring senescent cells as a strategy for reducing HPV risk of cervical cancer development and identifying the cell of tumor origin, which could be beneficial for improving the utility of senolytic agents and immunotherapy in clinical practice.

Mechanism of low-intensity pulse ultrasound combined with Rhodiola to promote bone formation in spinal fusion

Lanjun Zhang — 2024-10-08

This study aimed to explore the influence and mechanism of low-intensity pulsed ultrasound (LIPUS) combined with Rhodiola bone penetration on the formation of spinal fusion bone. Sixty clean-grade New Zealand white rabbits were selected for randomization and divided into combined group and Rhodiola group, with 30 rabbits in each group to construct a rabbit lumbar intervertebral fusion model, using Rhodiola intervention and Rhodiola combined with LIPUS intervention protocol, respectively. The axial strength, axial stiffness, maximum compressive load, vascular endothelial growth factor (VEGF), cycloxygenase-2 (COX-2), prostaglandin E2 (PGE2) and transforming growth factor-β (TGF-β) were compared after HE staining, immunohistochemistry and biomechanical detection. Spine fusion rate was 100.00%; the combined bone graft tissue had implanted bone cell degeneration, cell necrosis and cell hyperplasia, chondrocytes differentiated into trabecular bone and some hematopoietic cells, severe cell necrosis and fiber cell proliferation and late bone formation in the Rhodiola group, VEGF, COX-2, PGE2, TGF-β, axial strength, axial stiffness, and maximum compression load in the combined group significantly increased (P<0.05). Spinal fusion using LIPUS combined with Rhodiola can enhance biomechanical properties and promote the role of PGE2, COX-2, VEGF, TGF-β expression and bone formation, and this protocol is worthy of clinical application.

MTHFR C677T gene polymorphism in patients with coronary heart disease and hypertension treated with enalapril and folic acid: implications for prognosis

Li Ma — 2024-10-08

We aimed to investigate the effect of the methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism on the prognosis of patients with coronary heart disease (CHD) and hypertension treated with enalapril and folic acid. A total of 540 CHD patients diagnosed by coronary angiography in our hospital were selected. According to whether there was folic acid intervention, they were divided into a folic acid group, a non-folic acid group and a control group. The genotypes of the MTHFR C677T locus were detected. Hcy concentration and the folate content were determined. In folic acid group, enalapril and folic acid tablets were used to reduce blood pressure, and clopidogrel or ticagrelor were selected according to CYP2C19 genotypes. In non-folic acid group, enalapril tablets were used, and clopidogrel or ticagrelor were selected based on CYP2C19 genotyping. Routine treatment without intervention was used in control group. Patients were prescribed standardized drug treatment and were followed up by an outpatient service or by telephone for 12 months after discharge. We found that the number and proportion of MTHFR C677T gene mutations in the folic acid group, non-folic acid group and control group were 150 (83.3%), 142 (78.9%) and 144 (80.0%), respectively. The recurrence rate of cardiovascular events in the folic acid and non-folic acid groups was significantly lower, and the degree of reduction in the recurrence rate of cardiovascular events in the folic acid and non-folic acid groups was significantly different. The concentrations of TG, TC, and LDL-C in folate and non-folic groups were lower, while HDL-C was higher than that in control group. To sum up, screening high-risk populations with genotypes has great significance in improving the clinical outcome of CHD patients with H-type hypertension. Folic acid supplementation improves the compliance rate of patients’ blood pressure levels and improves their clinical prognosis as well.

LC-QTOF-MS/MS metabolic profiling and hepatoprotective effects of Litsea monopetala bark methanol extract against liver injury in rats and HepG2 cells

Subhasish Sahoo — 2024-10-08

Fresh stem bark decoction of Litsea monopetala has been practiced for the treatment of jaundice and other liver disorders by the tribal communities of Thakht-e-Sulaiman hills from West Pakistan. As per the folkloric claim, this study aims to identify the phytoconstituents and evaluate the hepatoprotective action of stem bark methanol extract of L. monopetala (LMME). The in-vitro hepatoprotective effect of L. monopetala was performed by H₂O₂-induced toxicity in the HepG2 cell line and in-vivo by cclt;sub>4-induced hepatotoxicity in Wistar albino rats taking Silymarin as standard drug. Phytoconstituents were identified using LC-QTOF-MS analysis followed by in-silico docking and validation. Molecular docking interactions between identified compounds of L. monopetala and two target proteins, namely 1VJY and 5HYK were presented. In this study, treatment with LMME at 100 µg/mL showed 67.73 % cell viability as compared to H₂O₂ (100 µM) treated alone i.e., 18.55 % in the HepG2 cell line. In-vivo treatment of LMME reversed the altered serum biochemical parameters and reduced the inflammatory response similar to that of the Silymarin-treated group supported by histopathological investigation. This research reveals that L. monopetala is a rich source of flavonoids and phenols which supports its hepatoprotective effects and is proposed for its usage as a promising hepatoprotective agent after controlled trials

Methyl jasmonate effects on Lactuca serriola L.: Antioxidant defense and bioactive compound changes

Maisa Asheri — 2024-10-08

The effect of methyl jasmonate (MeJA) foliar spray on the activity of antioxidant enzymes—Superoxide dismutase (SOD), Catalase (CAT), Ascorbate peroxidase (APX), and Guaiacol peroxidase (GPX)—along with assessments of total phenolic and flavonoid contents and antioxidant activity (IC50), was examined in Prickly lettuce (Lactuca serriola L.). The study involved treating plants with three MeJA solutions (0, 200, and 400 µM) and harvesting samples at four distinct time intervals. Varied MeJA concentrations and time intervals resulted in a substantial increase in the activity of all the antioxidant enzymes investigated in this study. Both concentration levels and time courses exhibited progressive outcomes. Moreover, MeJA treatment led to elevated levels of total phenolic and flavonoid contents, reaching peaks of 17.02 (mg GAL/g DW) and 8.3 (mg QUE/g DW), respectively, particularly in response to the 400 µM concentration. However, the total flavonoid content did not show any significant variation between the two concentrations. Based on the half-maximal inhibitory concentration (IC50) values, the antioxidant activity in MeJA-treated plants was found to be lower compared to the controls. However, our findings suggest that, under specific conditions discussed in this study, MeJA has the potential to enhance the nutritional value of L. serriola.

Clinicopathological characteristics of SMAD4 gene expressions in colorectal cancer patients

Neda Mansouri — 2024-10-08

Colorectal cancer (CRC) ranks as the third leading cause of cancer-related deaths globally. SMAD4 gene acts as the central mediator of the signaling pathway for transforming growth factor-β (TGFβ) with a significant effect on colorectal cancer. Previous research has confirmed a relationship between the presence of the SMAD4 gene and the survival and progression of colorectal cancer in patients. In this study, our goal was to analyze the presence of SMAD4 in both colorectal cancer and nearby normal tissues. The expression levels of SMAD4 were evaluated in 45 colorectal tumor tissues and 45 adjacent control tissues using the Quantitative Real-Time PCR (qRT-PCR) method. Additionally, we assessed the diagnostic effectiveness of SMAD4 by creating a receiver operating characteristic (ROC) curve. Our findings showed that the expression of SMAD4 was significantly reduced in colorectal cancer patients compared to the adjacent control group sample. Examination of clinicopathological characteristics of patients revealed varied correlations between SMAD4 gene expressions and TMN stage (p<0.0001). These findings suggest that SMAD4 levels could be used as possible diagnostic indicators for colorectal cancer.

Mitochondrial DNA sequence-based identification of two subterranean termite species, from Riyadh Province, Kingdom of Saudi Arabia

Rasool Khawaja Ghulam — 2024-10-08

Termites are economically important wood-destroying and agricultural pests. The termite fauna almost consists of 2900 described species in 286 genera worldwide. In the present study, hundreds of termite samples from 42 different locations in the Riyadh province were collected. These samples were previously used for morphometric identification and reported two subterranean termite species, Coptotermes heimi and Psammotermes hypostoma, in the family Rhinotermitidae. In the present study, these samples were analysed using DNA barcoding with the mitochondrial cytochrome c oxidase subunit 1 gene to confirm the conventional taxonomical identification on a molecular basis. The obtained COI gene sequences of all 42 termite specimens were submitted to GenBank (accession numbers: ON529959-ON529969, OP825131-OP825132, and OP890882-OP890910). Eleven of the 42 samples were thus identified as C. heimi and the remaining 31 samples as P. hypostoma, which were phylogenetically analysed. All the 11 C. heimi sequences were grouped in a single clade, indicating close relatedness. While 31 sequences of P. hypostoma constituted two clades in the phylogenetic tree. Pairwise nucleotide sequence identity and divergence analysis showed that C. heimi sequences showed high nucleotide identities of 87.6-99.5% and less divergence ranging from 0.5% to 13.6%. Similarly, sequences of P. hypostoma also showed high nucleotide identity of 78.6-100% and low divergence among them ranging from 0-10.7%. A further application, significance, and shortcomings of COI-based DNA barcoding have been discussed. DNA barcoding using the COI gene is a reliable tool to distinguish C. heimi and P. hypostoma genotypes.

Pharmacological and economical aspects of important species of Cordyceps sensu lato: A review

Jameel M. Al-Khayri — 2024-10-08

Cordyceps is a well-known endo-parasitic fungus commonly used in traditional Chinese medicine for a long time. The demand for Cordyceps is increasing daily because it is commonly used as a nutritional food, medicine, and supplement owing to its natural source. It is very attractive in almost all countries with no side effects. Most Cordyceps species have been studied in China, Bhutan, India, Japan, South Korea, and Nepal. We have discussed the important contents of Cordyceps, dietary source and nutritional value of Cordyceps, multiple pharmacological properties of four important Cordyceps species, Ophiocordyceps sinensis, Cordyceps militaris, Cordyceps cicadae, and Cordyceps tenuipes, along with the economic status of Cordyceps and its benefits in terms of medicine, supplements, and the cosmetics industry. Owing to the high demand and several benefits of Cordyceps, it offers mysterious economic improvements in developed and underdeveloped countries. Therefore, more attention is required to save the endangered species of Cordyceps to fulfil the medicinal and nutritional demands worldwide.

The role of metabolic disorders in the development of atherosclerosis

Alexander V. Blagov — 2024-10-08

Atherosclerosis is a major risk factor for cardiovascular disease (CVD), which is the leading cause of death worldwide. Atherosclerosis is initiated by endothelial activation, followed by a cascade of events (accumulation of lipids, fibrous elements, and calcification) triggering vasoconstriction and activation of inflammatory pathways. This review focuses on the various stages in the development of atherosclerosis, ranging from endothelial dysfunction to plaque rupture. In addition, disorders of lipid, glucose and amino acid metabolism in atherosclerosis are considered here. The key pathological stages of metabolism disruption and their role in atherosclerosis are considered in detail which may be helpful for the more better understanding of atherosclerosis pathogenesis. Finally, some therapeutic approaches aimed at modulating lipid metabolism will also be presented which show the therapeutic targets (enzymes and transport proteins) which modulation can prevent further deterioration of patients symptoms

The role of NETosis in enhancing of atherosclerosis

Alexander V. Blagov — 2024-10-08

Activated neutrophils release neutrophil extracellular traps (NETs), complex structures composed of extracellular genetic material and proteins sourced from the nucleus, granules, and cytoplasm in response to pathogenic inflammatory conditions. These NETs play a crucial role in the host's innate immune defense against invasive infections. Notably, in conditions like atherosclerosis, these extracellular formations can also be elicited by inflammatory stimuli such as lipids, prothrombotic factors, platelet aggregation, or proinflammatory cytokines. NETs have been identified on the inner arterial walls in cardiovascular disease states. By promoting inflammation through NETosis-mediated cell adhesion processes and exerting cytotoxic effects leading to cellular dysfunction and tissue damage, NETs contribute to the pathogenesis of inflammatory conditions.