Anti-cytokine Storm Activity of Fraxin, Quercetin, and their Combination on Lipopolysaccharide-Induced Cytokine Storm in Mice: Implications in COVID-19.

Background: Cytokine release syndrome (CRS) is the leading cause of mortality in advanced stages of coronavirus patients. This study examined the prophylactic effects of fraxin, quercetin, and a combination of fraxin+quercetin (FQ) on lipopolysaccharide-induced mice. Methods: Sixty mice were divided into six groups (n=10) as follows: control, LPS only, fraxin (120 mg/Kg), quercetin (100 mg/Kg), dexamethasone (5 mg/Kg), and FQ. All treatments were administered intraperitoneally (IP) one hour before induction by LPS (5 mg/Kg) IP injection. Twenty-four hours later, the mice were euthanized. Interleukin one beta (IL-1ß), interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-α) were quantified using an enzyme-linked immunosorbent assay (ELISA), and lung and kidney tissues were examined for histopathological alterations. This study was conducted at Al-Nahrain University, Baghdad, Iraq, in 2022. Results: FQ reduced IL-1ß (P<0.001). All treatments significantly suppressed IL-6, fraxin, quercetin, dexamethasone, and FQ, all with P<0.001. The TNF-α level was reduced more with dexamethasone (P<0.001) and quercetin (P<0.001). Histopathological scores were significantly reduced mainly by quercetin and FQ in the lungs with scores of 12.30±0.20 (P=0.093), and 15.70±0.20 (P=0.531), respectively. The scores were 13±0.26 (P=0.074) and 15±0.26 (P=0.222) for quercetin and FQ in the kidneys, respectively. Conclusion: All used treatments reduced proinflammatory cytokine levels and protected against LPS-induced tissue damage.

The impact of methamphetamine on psychosocial variables in patients from Iraq.

Background: The current work reviews the psychosocial factors associated with different urinary methamphetamine concentration levels. Methods: From April to November 2023, 243 participants from Baghdad's Al-Ataa Hospital were the subjects of a cross-sectional descriptive analysis study. We included 73 patients in this study. Result: The urinary methamphetamine concentration levels were from 3 to 92,274 ng/ml, with a mean ± SD of 10,873.6 ± 18,641. Patients diagnosed with major depression disorder exhibited higher scores on GHQ-30, UCLA, MOAS, and BDI-II with a significant P-value of 0.0001, 0.001, 0.0001, and 0.0001, consequently with an effect size of 0.015, 0.001, 1.05, and 3.24, respectively. Conclusions: The multi-screening test can produce a false positive. It frequently interferes with other drugs, especially antidepressants. This will result in patients being stigmatized and accused. On the other hand, those who accidentally come into contact with crystal smoke will experience the same withdrawal symptoms as the addicted patients. Their urinary methamphetamine level (titer) could have negative results. Urinary methamphetamine levels should be zero in healthy patients. In this situation, screening tests, expert opinion, and urine methamphetamine testing are strongly recommended.

Papaverine Enhances the Oncolytic Effects of Newcastle Disease Virus on Breast Cancer In Vitro and In Vivo.

Breast cancer is a lethal disease in females worldwide and needs effective treatment. Targeting cancer cells with selective and safe treatment seems like the best choice, as most chemotherapeutic drugs act unselectively. Papaverine showed promising antitumor activity with a high safety profile and increased blood flow through vasodilation. At the same time, it was widely noticed that virotherapy using the Newcastle disease virus proved to be safe and selective against a broad range of cancer cells. Furthermore, combination therapy is favorable, as it attacks cancer cells with multiple mechanisms and enhances virus entrance into the tumor mass, overcoming cancer cells' resistance to therapy. Therefore, we aimed at assessing the novel combination of the AMHA1 strain of Newcastle disease virus (NDV) and nonnarcotic opium alkaloid (papaverine) against breast cancer models in vitro and in vivo. Methods. In vitro experiments used two human breast cancer cell lines and one normal cell line and were treated with NDV, papaverine, and a combination. The study included a cell viability MTT assay, morphological analysis, and apoptosis detection. Animal experiments used the AN3 mouse mammary adenocarcinoma tumor model. Evaluation of the antitumor activity included growth inhibition measurement; the immunohistochemistry assay measured caspase protein expression. Finally, a semiquantitative microarray assay was used to screen changes in apoptotic proteins. In vitro, results showed that the combination therapy induces synergistic cytotoxicity and apoptosis against cancer cells with a negligible cytotoxic effect on normal cells. In vivo, combination treatment induced a significant antitumor effect with an obvious regression in tumor size and a remarkable and significant expression of caspase-3, caspase-8, and caspase-9 compared to monotherapies. Microarray analysis shows higher apoptosis protein levels in the combination therapy group. In conclusion, this study demonstrated the role of papaverine in enhancing the antitumor activity of NDV, suggesting a promising strategy for breast cancer therapy through nonchemotherapeutic drugs.

Antiangiogenic Activity of Quinine Alone and in Combination with vitamin C in both ex vivo and in vivo Assays.

BACKGROUND: Angiogenesis is the process of vascularization from preexisting blood vessels. It is essential for many physiological and pathological processes. Quinine is an anti-malarial agent belongs to the quinoline alkaloid that can inhibit angiogenesis. Vitamin C is also an important antioxidant and has been shown to reduce angiogenesis in tumor. OBJECTIVE: The study was aimed at investigating the effect of quinine alone and in combination with vitamin C on angiogenesis process. MATERIALS AND METHODS: 12 to 14 weeks old male albino rats were used for the study. Quinine was prepared by dissolving in DMSO and was serially diluted. The rat aorta ring assay was employed to investigate the antiangiogenic effect of quinine ex vivo. An in vivo chorioallantoic membrane (CAM) assay was used to measure the blood vessels inhibition zone by quinine. The zone of inhibition was calculated as the mean inhibition area of a blood vessel in mm±SD.The obtained data were statistically analyzed. RESULTS: The results revealed that quinine has a significant dose-dependent inhibition effect on the growth of blood vessels by 98% ± 0.07 in concentration 100µg/ml when compared to the negative control. moreover, the inhibition of blood vessels growth as a measure of the antiangiogenic activity of quinine in combination with vitamin C shows a synergistic effect when the concentration that inhibit 50% of blood vessels growth (IC50) which equals to 5.05 µg/ml resulted in 85% of growth inhibition when combined with IC50 of vitamin C which equals to 22..87µg/ml. CONCLUSION: The findings suggest that the activity of quinine with vitamin C synergism can greatly lower blood vessels growth in rat aorta rings and CAM assays. Quininehas an inhibitory effect on tumor and can be utilized as an antiangiogenic agent alone or in combination with vitamin C.

The Effects of Abscisic Acid on Angiogenesis in Both ex vivo and in vivo Assays.

BACKGROUND: Angiogenesis is a complex biological process wherein novel capillary blood vessels mature from pre-existing vasculature for delivering tissues with oxygen and nutrients. Natural molecules that have anti-angiogenic activity and toxicity can raise the focus on using plant sources as essential therapeutic agent. OBJECTIVE: The current research was intended to estimate the probable anti-angiogenic activity of abscisic acid alone and in combination with prednisolone, a well-known angiostatic glucocorticoid. METHODS: two months old albino rats were used in this study. ABA and prednisolone stock solutions were prepared and added after embedding aortic rings in growth media. The ex vivo rat aorta ring assay (RAR) was applied to explore the anti-angiogenic effect of ABA. The in vivo chorioallantoic membrane assay (CAM) was applied to quantify the blood vessels inhibition zone by ABA effect. That zone was calculated as the mean inhibition region on eggs in mm ± SD. RESULTS: Abscisic acid screened byex vivo and in vivo assays, revealed a significant dose-dependent blood vessels inhibition in comparison to the negative control with IC50= 7.5µg/ml and a synergism effect when combined with prednisolone. CONCLUSION: The synergism activity of ABA with prednisolone can significantly inhibit blood vessels growth in RAR and CAM assays. These results shed the light on the potential clinic values of ABA, and prednisolone combination in angiogenesis-dependent tumors.

The Anti-Angiogenic and Anti-Proliferative Activity of Methyl Hydroxychalcone.

OBJECTIVE: Angiogenesis plays a key role in tumor growth, invasion, and metastasis of cancer diseases, and therefore, the inhibition of angiogenesis can provide an important therapeutic approach in cancer diseases. The aim of this study was to investigate the inhibitory effects of methyl hydroxychalcone on ex vivo sprouting of rat aortic micro-vessels and in vivo formation of chorionic plexus in chick chorioallantoic membrane and to investigate the mechanism underlying anti-angiogenic activity. METHODS: Rat aortic rings were sectioned by 1 mm. 500µl of 3 mg/ml of fibrinogen in serum free M199 growth medium was added to each well with 5 ug/ml of aprotinin. Methyl hydroxychalcone at varying concentrations ranging from 6.25 µg/ml to 100 µg/ml was added to the complete growth medium. Fertilized chicken eggs were incubated at 37°C. On day 3, a small window was opened in the shell. The window was sealed with adhesive tape and incubated until day 5. One mg of methylhydroxychalcone was applied. Images of each CAM were captured using a digital camera, and the dimensions of the blood vessels were measured digitally. Vascular endothelial growth factor (VEGF)-induced human umbilical vein endothelial cell (HUVEC) proliferation and tube formation assays were examined. Additionally, VEGF-165 levels and expression of membrane VEGF receptor-2 in HUVEC lysates have been assessed. RESULTS: The data showed that methyl hydroxychalcone significantly had antiangiogenic activity in a dose dependent manner in the rat aorta assay and had significant perturbation activity on blood vessels in the CAM assay. Methyl hydroxychalcone significantly inhibited proliferation and capillary-like tube formation in VEGF-induced HUVEC. Moreover, methylhydroxychalcone significantly reduced VEGF-165 levels in HUVECs lysate. CONCLUSION: This study showed that methyl hydroxychalcone significantly inhibits the angiogenesis process, blocking the VEGF signaling pathway in HUVECs and could be a potential promising angiogenesis inhibitor lead compound.

The Anti-Cytokine Storm Activity of Quercetin Zinc and Vitamin C Complex.

Cytokine storm is one of the causative deaths in a patient with severe acute respiratory syndrome. This study aimed at evaluating the prophylaxis effect of quercetin complexes with zinc and buffered ascorbic acid upon cytokine storm induction in mice and identifying the complex's acute toxicity. Mice were randomly divided into three groups: group A, control group, received 0.9% normal saline; group B received 100 mg/kg of the complex one hour before lipopolysaccharide (LPS) administration; and group C received the LPS IP 5 mg/kg. Then, levels of interleukin 1 and interleukin 6 were measured in the serum, and lung and kidney tissues were investigated for any changes that may have happened. Thirty mice were used to investigate the acute toxicity; mice were distributed into six groups: one control group and five treated groups; then several serial dilutions from the complex have been prepared for different concentrations from 5 g/kg to 0.312 g/kg. The animals were observed for 14 days. The LD50 was deduced by the straight-line equation calculated from the dose-response curve. The results in this study showed that group A had no significant tissue change. LPS group C showed tissue damage in the lung and kidney, which significantly prevented by the pretreated complex in group B. Moreover, the complex's acute toxicity value (LD50) was 655 mg/kg. In conclusion, the complex has significantly ameliorated LPS-induced acute lung and kidney injury, largely through suppression of inflammation; the large lethal dose value may make the complex have a promising therapeutic effect in the prevention of cytokine storm.

The Antiangiogenic Activity of Flaxseed Oil Alone and Combination with Mefenamic Acid in Vivo and in Vitro Assay.

BACKGROUND: Antiangiogenic agents are commonly used for the management of many types of cancers. Flaxseed oil is a wealthy source of omega-3,omega-6, and lignans, and possesses anticancerous and antioxidant activities. OBJECTIVE: To investigate the antiangiogenic activity of flaxseed oil alone and in combination with mefenamic acid in a dose-response study. METHODS: Oil was extracted from flaxseeds. The ex vivo rat aorta ring assay was used to screen the flaxseed oil alone and in combination with mefenamic acid for possible antiangiogenic activity. Also, the assay was used to determine the dose-response effect.An in vivo chick chorioallantoic membrane (CAM) assay was used to quantify the zone of inhibition of blood vessel by flaxseed oil. RESULTS: The 200,100,50,25,12.5, and 6.25µg/ml of the flaxseed oil inhibited blood vessel growth with values of 91±0.42, 84±2.06,36±2.71, 21±2.15,23±1.56, and 5±0.93%, respectively, withan IC50 value of 45.695 µg/ml. These concentrations showed a dose-dependent inhibition of angiogenesis. At 200, 100, and 50 µg/ml of flaxseed oil in combination with 50 µg/ml of mefenamic acid inhibited blood vessel growth with inhibition percentages of 81.48±0.82,79.63±0.75, and 77.78±1.26, respectively, with an IC50 of 2.27 µg/ml. Also, these concentrations indicated a dose-dependent inhibition of angiogenesis.  Flaxseed oil produced a significant inhibition zone of blood vessels in the CAM assay. The LD50 for flaxseed oil was 5.656g/kg. CONCLUSION: Flaxseed oil alone and in combination with mefenamic acid exhibited antiangiogenic activity both in ex vivo and in vivo assays. At doses of 2.5, 1.25, 0.625, and 0.312 g/kg of flaxseed oil intraperitoneally injected into mice, no symptoms of toxicity or death of mice due to acute toxicity were observed. However, doses of 5 and 10 g/kg of oil resulted in the death of the majority of mice.

Antiangiogenic Activity of Sweet Almond (Prunus dulcis) Oil Alone and in Combination with Aspirin in both in vivo and in vitro Assays.

Objective: Angiogenesis is new blood vessels formations that are necessary for certain physiological and pathologic conditions. Almond oil represents adjuvant therapies for numerous health benefits; It has ability for prevent the formation angiogenesis in tumour, due to its high concentration of unsaturated fatty acids, polyphenols, flavonoids and other ingredients content. Aspirin is non-steroidal ant-inflammatory drug; significantly reduce the angiogenesis of cancer. The aim of the study is to investigate the role of Prunus dulcis oil alone and in combination with aspirin, as an anti-angiogenic. Method: A sequential concentrations ex-vivo rat aorta ring assay, investigate the anti-angiogenic activity of Prunus dulcisoil in vivo. After three days of incubation, small holes were created on the fine pinpoint. Acute toxicity study was evaluated after administration of Prunus dulcis oil via intraperitoneal route. Results: the obtained results displayed that the serial concentration dose-response has a significant inhibition effect of blood vessels growth when compared to the negative control (DMSO 1%), the dose depended percentage of inhibition, sweet almond oil in combination with aspirin synergism effect to inhibition growth blood vessels as anti-angiogenic activity. The zone of inhibition was been measured as the mean of inhibition area of blood vessel on eggs in millimetre (mm) ± standard deviation. Conclusion: Almond oil dose has inhibition effect on cancer and can be used as (anti-angiogenesis), the activity of almond oil with Aspirin synergism can significantly reduce blood vessel's growth in rate aorta ring and CAM assay.

Anti-proliferation effects of benzimidazole derivatives on HCT-116 colon cancer and MCF-7 breast cancer cell lines.

Benzimidazoles 1-4 were obtained using modified synthesis methods and studied for their ability to inhibit cell proliferation of colon cancer cell HCT-116 and breast cancer cell MCF-7 using MTT assays. In the HCT-116 cell line, benzimidazole 2 was found to have an IC50 value of 16.2 ± 3.85 µg/mL and benzimidazole 1 a value of 28.5 ± 2.91 µg/mL, while that for benzimidazole 4 was 24.08 ± 0.31 µg/mL. In the MCF-7 cell line, benzimidazole 4 had an IC50 value of 8.86 ± 1.10 µg/mL, benzimidazole 2 a value of 30.29 ± 6.39 µg/mL, and benzimidazole 1 a value of 31.2 ± 4.49 µg/mL. Benzimidazole 3 exerted no cytotoxicity in either of the cell lines, with IC50 values >50 µg/mL. The results suggest that benzimidazoles derivatives may have chemotherapeutic potential for treatment of both colon and breast cancers.