Advances in traditional herbal formulation based nano-vaccine for cancer immunotherapy: Unraveling the enigma of complex tumor environment and multidrug resistance.

Cancer is attributed to uncontrolled cell growth and is among the leading causes of death with no known effective treatment while complex tumor microenvironment (TME) and multidrug resistance (MDR) are major challenges for developing an effective therapeutic strategy. Advancement in cancer immunotherapy has been limited by the over-activation of the host immune response that ultimately affects healthy tissues or organs and leads to a feeble response of the patient's immune system against tumor cells. Besides, traditional herbal medicines (THM) have been well-known for their essential role in the treatment of cancer and are considered relatively safe due to their compatibility with the human body. Yet, poor solubility, low bio-availability, and lack of understanding about their pathophysiological mechanism halt their clinical application. Moreover, considering the complex TME and drug resistance, the most precarious and least discussed concerns for developing THM-based nano-vaccination, are identification of specific biomarkers for drug inhibitory protein and targeted delivery of bioactive ingredients of THM on the specific sites in tumor cells. The concept of THM-based nano-vaccination indicates immunomodulation of TME by THM-based bioactive adjuvants, exerting immunomodulatory effects, via targeted inhibition of key proteins involved in the metastasis of cancer. However, this concept is at its nascent stage and very few preclinical studies provided the evidence to support clinical translation. Therefore, we attempted to capsulize previously reported studies highlighting the role of THM-based nano-medicine in reducing the risk of MDR and combating complex tumor environments to provide a reference for future study design by discussing the challenges and opportunities for developing an effective and safe therapeutic strategy against cancer.

Cerebrovascular disease in COVID-19: a systematic review and meta-analysis.

Background: The association between COVID-19 and acute cerebrovascular disease (CVD) has not been explored extensively. New data has come to light which may change previous results. Methods: We queried the PubMed electronic database from its inception until February 2022 for studies evaluating the incidence of stroke in COVID-19 patients. Results of the analysis were pooled using a random-effects model and presented as odds ratios (ORs) with 95% confidence intervals (95% CIs). Results: 37 studies consisting of 294,249 patients were included in our analysis. Pooled results show that the incidence of acute CVD events in COVID-19 positive patients is 2.6% (95% CI: 2.0-3.3; P<0.001). Cardioembolic (OR=14.15, 95% CI: 11.01 to 18.19, P<0.00001) and cryptogenic (OR=2.87, 95% CI: 1.91 to 4.32, P<0.00001) etiologies were associated with COVID-19 positivity. Risk factors for CVD events in patients with COVID-19 were atrial fibrillation (OR=2.60, 95% CI: 1.15 to 5.87, P=0.02), coronary artery disease (OR=2.24, 95% CI: 1.38 to 3.61, P=0.0010), diabetes (OR=2.46, 95% CI: 1.36 to 4.44, P=0.003) and hypertension (OR=3.65, 95% CI: 1.69 to 7.90, P=0.005). Conclusion: COVID-19 infection is associated with an increased risk for acute CVD and is associated with cardioembolic and cryptogenic etiologies and the risk factors of atrial fibrillation, coronary artery disease, diabetes and hypertension in COVID-19 positive patients.

Title: Immunotherapy; a ground-breaking remedy for spinal cord injury with stumbling blocks: An overview.

Spinal cord injury (SCI) is a debilitating disorder with no known standard and effective treatment. Despite its ability to exacerbate SCI sequel by accelerating auto-reactive immune cells, an immune response is also considered essential to the healing process. Therefore, immunotherapeutic strategies targeting spinal cord injuries may benefit from the dual nature of immune responses. An increasing body of research suggests that immunization against myelin inhibitors can promote axon remyelination after SCI. However, despite advancements in our understanding of neuroimmune responses, immunoregulation-based therapeutic strategies have yet to receive widespread acceptance. Therefore, it is a prerequisite to enhance the understanding of immune regulation to ensure the safety and efficacy of immunotherapeutic treatments. The objective of the present study was to provide an overview of previous studies regarding the advantages and limitations of immunotherapeutic strategies for functional recovery after spinal cord injury, especially in light of limiting factors related to DNA and cell-based vaccination strategies by providing a novel prospect to lay the foundation for future studies that will help devise a safe and effective treatment for spinal cord injury.

Mesenchymal stem cells to treat female infertility; future perspective and challenges: A review.

Infertility negatively impacts the overall health and social life of affected individuals and couples. Female infertility is their inability to perceive pregnancy. To date, polycystic ovary syndrome, primary ovarian insufficiency, fallopian tube obstruction, endometriosis, and intrauterine synechiae have been identified as the primary causes of infertility in women. However, despite the mutual efforts of clinicians and research scientists, the development of an effective treatment modality has met little success in combating female infertility. Intriguingly, significant research has demonstrated mesenchymal stem cells as an optimal source for treating infertility disorders. Therefore, here we attempted to capsulize to date available studies to summarize the therapeutic potential of mesenchymal stem cells in combating infertility in women by focusing on the underlying mechanism through which stem cells can reduce the effects of ovarian disorders. Furthermore, we also discussed the preclinical and clinical application of stem cell therapy, their limitation, and the future perspective to minimize these limitations.

Ischemic Brain Stroke and Mesenchymal Stem Cells: An Overview of Molecular Mechanisms and Therapeutic Potential.

Ischemic brain injury is associated with a high rate of mortality and disability with no effective therapeutic strategy. Recently, a growing number of studies are focusing on mesenchymal stem cell-based therapies for neurodegenerative disorders. However, despite having the promising outcome of preclinical studies, the clinical application of stem cell therapy remained elusive due to little or no progress in clinical trials. The objective of this study was to provide a generalized critique for the role of mesenchymal stem cell therapy in ischemic stroke injury, its underlying mechanisms, and constraints on its preclinical and clinical applications. Thus, we attempted to present an overview of previously published reports to evaluate the progress and provide molecular basis of mesenchymal stem cells (MSCs) therapy and its application in preclinical and clinical settings, which could aid in designing an effective regenerative therapeutic strategy in the future.

Regenerative medicine and traumatic brain injury: from stem cell to cell-free therapeutic strategies.

Traumatic brain injury (TBI) is a serious health concern, yet there is a lack of standardized treatment to combat its long-lasting effects. The objective of the present study was to provide an overview of the limitation of conventional stem-cell therapy in the treatment of TBI and to discuss the application of novel acellular therapies and their advanced strategies to enhance the efficacy of stem cells derived therapies in the light of published study data. Moreover, we also discussed the factor to optimize the therapeutic efficiency of stem cell-derived acellular therapy by overcoming the challenges for its clinical translation. Hence, we concluded that acellular therapy possesses the potential to bring a breakthrough in the field of regenerative medicine to treat TBI.

Lay abstract Traumatic brain injury (TBI) is a devastating disorder that can lead to lifelong disability. Due to the complexity of TBI, to date, there is no effective and specific treatment available, other than preventive measures or intensive care to minimize the rate of mortality. Though increasingly evident studies have reported the effective role of stem cells therapy to treat TBI. However, direct exposure to stem cells has its limitation. Therefore, this study aimed to present an overview of the challenges associated with commonly used stem-cell therapy and to discuss and establish the ground for novel cell-free therapy in light of previously reported research. We hope this review will help to understand the role of cell-free therapy stem cells in the treatment of head injury and pave the way for the development of future treatments.

Awareness, knowledge, and practice regarding to diabetic retinopathy among KKU students besides medical students in Abha, Saudi Arabia.

BACKGROUND: Diabetes mellitus (DM) is a global public health problem. Global prevalence of diabetes is 8.5% in adult population. The prevalence of diabetic retinopathy (DR) is increasing day by day, the number of persons with diabetes will double by 2030. It is a serious cause of irreversible blindness and is the most common complication of diabetes. Annual fundus examination for diabetics aids in the prevention of blindness and allows intervening at a timely manner. This study's intent to estimate and improve level of awareness (A), knowledge (K), and practice (P) among all King Khalid University (KKU) students besides medical students in Abha, Saudi Arabia. METHODS AND MATERIALS: This is a cross-sectional survey that targets all KKU students besides medical students in Abha, Saudi Arabia. The researchers will use closed-end questions for awareness (A), knowledge (K), and practice (P). The data and the questionnaires will be sent to the sample by social media. The data will be analyzed by statistical package for the social sciences program (SPSS). RESULTS: A total of 635 KKU students completed the questionnaire. Female students were more than male students, 334 (52.6%) for females and 301 (47.4%) for males, respectively. Ages ranged from 18 to 24 years with a mean 23 ± 2 years. There was a good awareness for some of the factors related to the DR which is noted in the results. Awareness of smoking and pregnancy rate is extremely low compared to the rest of the factors related to the DR. CONCLUSION: There was high awareness regarding DR and its risk factors among KKU students but low awareness regarding smoking and pregnancy relationship with DR. Improvement is required for smoking and pregnancy with the progression DR.

Conformational turn triggers regio-selectivity in the bioactivation of thiophene-contained compounds mediated by cytochrome P450.

In the present work, we performed Density Functional Theory calculations to explore the bioactivation mechanism of thiophene-containing molecules mediated by P450s. For this purpose, relatively large size compounds, 2,5-diaminothiophene derivatives were selected particularly for this investigation. Here we found the successive regio-selectivity triggered by conformational turn played a significant role in the occurrence of bioactivation. 2,5-Diaminothiophene was oxidized to a 2,5-diimine thiophene-reactive intermediate by Compound I (Cpd I) through successive activations of two N-H bonds (H3-N11 and H1-N6). This reaction exhibited three special characteristics: (1) self-controlled regio-selectivity during the oxidation process. There was a large scale of conformational turn in the abstraction of the first H atom which triggers the selection of the second H for abstraction. (2) Proton-shuttle mechanism. In high spin (HS) state, proton-shuttle mechanism was observed for the abstraction of the second H atom. (3) Spin-selective manner. In protein environment, the energy barrier in HS state was much lower than that in low spin state. The novel proposed bioactivation mechanism of 2,5-diaminothiophene compounds can help us in rational design of thiophene-contained drugs avoiding the occurrence of bioactivation.

Correction to: CBFA2T2 is associated with a cancer stem cell state in renal cell carcinoma.

[This corrects the article DOI: 10.1186/s12935-017-0473-z.].

CBFA2T2 is associated with a cancer stem cell state in renal cell carcinoma.

BACKGROUND: Renal cell carcinoma (RCC) is the most common kidney cancer, accounting for approximately 80-90% of all primary kidney cancer. Treatment for patients with advanced RCC remains unsatisfactory. Rare cancer stem cells (CSCs) are proposed to be responsible for failure of current treatment. METHODS: OncoLnc was used as a tool for interactively exploring survival correlations. Gene manipulation and expression analysis were carried out using siRNA, RT-PCR and Western blotting. Wound healing and invasion assays were used for phenotypical characterization. Aldefluor assay and FACS sorting Sphere culture were used to determine the "stemness" of CSCs. Co-Immunoprecipitation (Co-IP) was used to examine the interaction between OCT4 and CBFA2T2. Student's t-test and Chi square test was used to analyze statistical significance. RESULTS: CBFA2T2 expression can significantly predict the survival of RCC patients. Knocking-down of CBFA2T2 can inhibit cell migration and invasion in RCC cells in vitro, and reduce ALDHhigh CSCs populations. CBFA2T2 expression is necessary for sphere-forming ability and cancer stem cells marker expression in RCC cell lines. CONCLUSIONS: Our data suggest that CBFA2T2 expression correlates with aggressive characteristics of RCC and CBFA2T2 is required for maintenance of "stemness" through regulation of stem cells factors, thereby highlighting CBFA2T2 as a potential therapeutic target for RCC treatment.

Furosine induces DNA damage and cell death in selected human cell lines: a strong toxicant to kidney Hek-293 cells.

Ne-(2-furoylmethyl)-l-lysine (furosine) is well-known indicator of early stage of Maillard reaction in processed food. Yet the toxicological aspects associated with its exposure remain rarely studied. Here, we investigated the effects of furosine exposure on cell viability, DNA damage, and its mutagenic potential by using MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide), TUNEL assay (Terminal deoxynucleotidyl transferase mediated dUTP Nick End Labeling assay), and Ames assay techniques on human cell lines, i.e., liver HepG-2, kidney Hek-293, neuronal SK-N-SH, and intestinal Caco-2, respectively. Our results showed that kidney Hek-293 cell line was the most sensitive to furosine exposure as significant reduction in cell viability and induction of DNA damage were observed at 50 mg/L concentration. In contrast, intestinal Caco-2 cell lines showed resistance to furosine exposure as DNA damage was only observed at 800 mg/L concentration of furosine. Ames assay indicated that furosine has no mutagenic effects on TA 100 and TA 1535 strains. Hence, this study suggests that furosine is a strong toxicant for kidney cells.

Short communication: Influence of preserving factors on detection of ß-lactamase in raw bovine milk.

The refrigeration (4, 8, 24, or 48 d), freezing (1, 7, or 30 h), thawing (25, 40, or 60°C), and usage of preservatives (sodium azide, potassium dichromate, sodium thiocyanate, bronopol, and methanol), of raw bovine milk were investigated for ß-lactamase activity. The ß-lactamase activity in all samples was assessed by using the test strips, and the samples with the preservatives were further assessed for ß-lactamase activity using the cylinder plate method. Results showed that the refrigeration, freezing, and thawing of raw bovine milk samples had no influence on the assay of the ß-lactamase activity. The addition of sodium azide or potassium dichromate as a sample preservative failed the test of ß-lactamase activity, whereas the addition of sodium thiocyanate, bronopol, or methanol had no influence on the test. In conclusion, sodium azide and potassium dichromate were not suitable preservatives for assessing ß-lactamase activity in raw milk when the cylinder plate method was used.

Identification of protein complexes of microsomes in rat adipocytes by native gel coupled with LC-ESI-QTOF.

The study of the composition of microsome proteins/complexes/interactions in adipocytes provides useful information for researchers related to energy metabolism disorders. The native gel coupled with LC-ESI-QTOF approach was employed here for separating protein complexes. We found a series of proteins functionally clustered in biological processes of protein metabolism, cellular carbohydrate catabolism, response to stimulus and wounding, macromolecular complex subunit organization, positive regulation of molecular function, regulation of programmed cell death and biomolecule transport. According to clustering of proteins' electrophoresis profiles across native gel fractions and bioinformatics data retrieval, protein complexes/interactions involved in protein metabolism, cellular carbohydrate catabolism, macromolecular complex subunit organization and biomolecule transport were identified. Besides, the results also revealed some functional linkages, which may provide useful information for discovering previously unknown interactions. The interaction between SSAO and ALDH2 was verified by co-immunoprecipitation. The native gel combining mass spectrometry approach appeared to be a useful tool for investigating microsome proteins and complexes to complement the traditional electrophoresis approaches. The native gel strategy together with our findings should facilitate future studies of the composition of rat adipocyte microsome protein complexes under different conditions.

Mechanistic Study of Silver Nanoparticle's Synthesis by Dragon's Blood Resin Ethanol Extract and Antiradiation Activity.

Biological synthesis of nanoparticles is best way to avoid exposure of hazardous materials as compared to chemical manufacturing process which is a severe threat not only to biodiversity but also to environment. In present study, we reported a novel method of finding antiradiation compounds by bioreducing mechanism of silver nanoparticles formation using 50% ethanol extract of Dragons blood, a famous Chinese herbal plant. Color change during silver nanoparticles synthesis was observed and it was confirmed by ultra violet (UV) visible spectroscopy at wave length at 430 nm after 30 min of reaction at 60 °C. Well dispersed round shaped silver nanoparticles with approximate size (4 nm to 50 nm) were measured by TEM and particle size analyser. Capping of biomolecules on Ag nanoparticles was characterized by FTIR spectra. HPLC analysis was carried out to find active compounds in the extract. Furthermore, antiradiation activity of this extract was tested by MTT assay in vitro after incubating the SH-SY5Y cells for 24 h at 37 °C. The results indicate that presence of active compounds in plant extract not only involves in bioreduction process but also shows response against radiation. The dual role of plant extract as green synthesis of nanoparticles and exhibit activity against radiation which gives a new way of fishing out active compounds from complex herbal plants.

Astroglial U87 Cells Protect Neuronal SH-SY5Y Cells from Indirect Effect of Radiation by Reducing DNA Damage and Inhibiting Fas Mediated Apoptotic Pathway in Coculture System.

Recent studies provide the evidence that indirect effects of radiation could lead to neuronal cells death but underlying mechanism is not completely understood. On the other hand astroglial cells are known to protect neuronal cells against stress conditions in vivo and invitro. Yet, the fate of neuronal cells and the neuroprotective effect of coculture system (with glial cells) in response to indirect radiation exposure remain rarely discussed. Here, we purpose that the indirect effect of radiation may induce DNA damage by cell cycle arrest and receptor mediated apoptotic cascade which lead to apoptotic death of neuronal SH-SY5Y cells. We also hypothesized that coculture (with glial U87) may relieved the neuronal SH-SY5Y cells from toxicity of indirect effects radiation by reducing DNA damage and expression of apoptotic proteins in vitro. In the present study irradiated cell conditioned medium (ICCM) was used as source of indirect effect of radiation. Neuronal SH-SY5Y cells were exposed to ICCM with and without coculture with (glial U87) in transwell coculture system respectively. Various endpoints such as, cell survival number assay, Annexin V/PI assay, cell cycle analysis by flow cytometer, mRNA level of Fas receptor by q RT-PCR, expression of key apoptotic proteins by western blot and estimation of neurotrophic factors by ELISA method were analyzed into neuronal SH-SY5Y cells with and without co culture after ICCM exposure respectively. We found that ICCM induced DNA damage in neuronal SH-SY5Y cells by significant increase in cell cycle arrest at S-phase (***P < 0.001) which was further supported by over expression of P53 protein (**P < 0.01). While coculture (with glial U87), significantly reduced the ICCM induced cell cycle arrest and expression of P53 ((###) P < 0.001) neuronal SH-SY5Y cells. Further investigation of the underlying apoptotic mechanism revealed that in coculture system; ICCM induced elevated level of FAS mRNA level was significantly reduced ((###) P < 0.001) in neuronal SH-SY5Y cells which was followed by significant reduction in expression of key apoptotic protein i.e., FADD ((###) P < 0.001), caspase-8 ((###) P < 0.001), and cleaved caspase-3 ((###) P < 0.001) as compare to neuronal SH-SY5Y cells which received ICCM without coculture. Intriguingly, concentration of neurotrophic factors such as, GDNF and BDNF were significantly increased ((###) P < 0.001) in neuronal SH-SY5Y after coculture (with glial U87) cells. Hence, these findings infer that the receptor mediated pathway could be the one way through which indirect effects of radiation cause neurotoxicity. However, in co-cultures system (with glial U87) neuronal SH-SY5Y depicts remarkable resistance against ICCM induced neurotoxicity.

Glial U87 cells protect neuronal SH-SY5Y cells from indirect effect of radiation by reducing oxidative stress and apoptosis.

Recent studies have demonstrated the role of indirect effect of radiation in neurodegeneration. However, the role of glial cells in neuroprotection against indirect effect of radiation is still not clear, although they are known to protect neurons under stress conditions in central nervous system. Our study showed that indirect effect of radiation increased the oxidative stress that further enhances the expression of key apoptotic proteins and leads to neuronal cell death. We also investigated the indirect effect of radiation on neuronal cells in the presence of glial cells in a transwell co-culture system, while our analysis was focused on neuronal cells. Irradiated cell-conditioned medium (ICCM) was used as source of indirect radiation and neuroprotective effect was analyzed by various endpoints. It was observed that ICCM-induced reactive oxidative species level was significantly reduced in SH-SY5Y cells co-cultured with glial U87 cells, which might help to maintain the integrity of mitochondrial membrane potential. Increased levels of antioxidant enzyme superoxide dismutase and antioxidant glutathione were observed in SH-SY5Y cells co-cultured with glial U87 cells. Moreover, it was also observed that co-culture with glial cells inhibits the expression of ICCM-induced apoptotic proteins, i.e. Bax, cytochrome c, and caspase-3 in SH-SY5Y cells. Hence, it can be speculated that in co-culture system glial cells may protect the neuronal SH-SY5Y cells by reducing the ICCM-induced oxidative stress and apoptotic death.

Indirect effects of radiation induce apoptosis and neuroinflammation in neuronal SH-SY5Y cells.

Recent studies have evaluated the role of direct radiation exposure in neurodegenerative disorders; however, association among indirect effects of radiation and neurodegenerative diseases remains rarely discussed. The objective of this study was to estimate the relative risk of neurodegeneration due to direct and indirect effects of radiation. (60)Co gamma ray was used as source of direct radiation whereas irradiated cell conditioned medium (ICCM) was used to mimic the indirect effect of radiation. To determine the potency of ICCM to inhibit neuronal cells survival colony forming assay was performed. The role of ICCM to induce apoptosis in neuronal SH-SY5Y cells was estimated by TUNEL assay and Annexin V/PI assay. Level of oxidative stress and the concentration of inflammatory cytokines after exposing to direct radiation and ICCM were evaluated by ELISA method. Expression of key apoptotic protein following direct and indirect radiation exposure was investigated by western blot technique. Experimental data manifest that ICCM account loss of cell survival and increase apoptotic induction in neuronal SH-SY5Y cells that was dependent on time and dose. Moreover, ICCM stimulate significant release of inflammatory cytokines i.e., tumor necrosis factor TNF-alpha (P < 0.01), Interleukin-1 (IL-1, P < 0.001), and Interleukin-6 (IL-6, P < 0.001) in neuronal SH-SY5Y cells and elevate the level of oxidative stress (MDA, P < 0.01). Up-regulation of key apoptotic protein expression i.e., Bax, Bid, cytochrome C, caspase-8 and caspase-3 confirms the toxicity of ICCM to neuronal cells. This study provides the evidence that indirect effect of radiation can be as much damaging to neuronal cells as direct radiation exposure can be. Hence, more focused research on estimation risks of indirect effect of radiation to CNS at molecular level may help to reduce the uncertainty about cure and cause of several neurodegenerative disorders.

LX loaded nanoliposomes synthesis, characterization and cellular uptake studies in H2O2 stressed SH-SY5Y cells.

In this study, we report the cellular uptake studies of novel LX loaded nanoliposomes in H2O2 stress SH-SY5Y Cells synthesized by thin film evaporation method. We have isolated the smallest size nanoliposomes after 90 min ultrasonification, keeping Polydisperse Index as 0.259. The morphology, size, zepta potential and drug efficiency of prepared nanoliposomes are characterized by using Transmission Electron Microscope (TEM), particle size analyzer and High Pressure Liquid Chromatography (HPLC). The particle size analyzer have confirmed the particle size of nanoluposomes measured in range of 100-250 nm, whereas the shape of these nanoliposomes is almost spherical. The zeta potential of small size nanoliposomes was measured as -49.62 and encapsulation efficiency of the LX loaded nanoliposomes was 87%. The oxidative stress response in SH-SY5Y Cells for various doses of drug with and without nanoliposomes has affectively improved the cell-stress response up to 20% after 24 h of incubation at 37 degrees C. The results indicated that LX loaded nanoliposomes were taken by the cells effectively which ultimately improved the cell-stress response. Thus, this study confirmed that synthesized nanoliposomes are not only effective drug carriers but could be potentially used for delivery of genes, antibodies, and proteins in future.

Differential expression of specific cellular defense proteins in rat hypothalamus under simulated microgravity induced conditions: comparative proteomics.

Microgravity severely halts the structural and functional cerebral capacity of astronauts especially affecting their brains due to the stress produced by cephalic fluid shift. We employed a rat tail suspension model to substantiate simulated microgravity (SM) in brain. In this study, comparative mass spectrometry was applied in order to demonstrate the differential expression of 17 specific cellular defense proteins. Gamma-enolase, peptidyl-prolyl cis-trans isomerase A, glial fibrillary acidic protein, heat shock protein HSP 90-alpha, 10 kDa heat shock protein, mitochondrial, heat shock cognate 71 kDa protein, superoxide dismutase 1 and dihydropyrimidinase-related protein 2 were found to be upregulated by HPLC/ESI-TOF. Furthermore, five differentially expressed proteins including 60 kDa heat shock protein, mitochondrial, heat shock protein HSP 90-beta, peroxiredoxin-2, stress-induced-phosphoprotein, and UCHL-1 were found to be upregulated by HPLC/ESI-Q-TOF MS. In addition, downregulated proteins include cytochrome C, superoxide dismutase 2, somatic, and excitatory amino acid transporter 1 and protein DJ-1. Validity of MS results was successfully performed by Western blot analysis of DJ-1 protein. This study will not only help to understand the neurochemical responses produced under microgravity but also will give future direction to cure the proteomic losses and their after effects in astronauts.