Separation of crude methanol extract of Plantago major root: identification of phytochemicals and assessment of biological activities.

Plantago major root extracts were used for analysis by Gas Chromatography-Mass Spectrometry (GC-MS). The anticancer and antibacterial functions of extracts were also investigated. The dichloromethane extract of P. major had the highest inhibitory effect against Salmonella paratyphi (18.00 ± 1.4 mm) at 100 mg/mL concentration. The lowest MIC was also achieved for S. paratyphi treated with dichloromethane extract of P. major (1.5 mg/mL). The minimum MBC (2 mg/mL) was observed for dichloromethane extract of P. major root against S. paratyphi. IC50 values of dichloromethane extracts of P. major root (184.84 µg/mL) against HCT116 were lower than the ethyl acetate and butanol extracts (212.41 µg/mL and 223.93 µg/mL) at 72h. The butanol extract exhibited the most IC50 value on HEK293 (748.19 µg/mL). The biological properties of P. major extracts may be assigned to the presence of numerous compounds detected in GC/MS analysis including n-Hexadecanoic acid, Linolenic acid, Palmitic acid, methyl ester, Stearic acid.

HK-2 cell response to TGF-ß highly depends on cell culture medium formulations.

The immortalized human renal proximal tubular epithelial cell line HK-2 is most commonly used to study renal cell physiology and human kidney diseases with tubulointerstitial fibrosis such as diabetic nephropathy, obstructive uropathy or allograft fibrosis. Epithelial-to-mesenchymal transition (EMT) is the main pathological process of tubulointerstitial fibrosis in vitro. Transforming growth factor-beta (TGF-ß) is a key inducer of EMT. Several pro-fibrotic gene expression differences have been observed in a TGF-ß-induced EMT model of HK-2 cells. However, growth conditions and medium formulations might greatly impact these differences. We investigated gene and protein expression of HK-2 cells cultured in six medium formulations. TGF-ß1 increased the expression of ACTA2, TGFB1, COL4A1, EGR2, VIM and CTGF genes while reducing PPARG in all medium formulations. Interestingly, TGF-ß1 treatment either increased or decreased EGR1, FN, IL6 and C3 gene expression, depending on medium formulations. The cell morphology was slightly affected, but immunoblots revealed TGFB1 and vimentin protein overexpression in all media. However, fibronectin expression as well as the nuclear translocation of EGR1 was medium dependent. In conclusion, our study demonstrates that, using the HK-2 in vitro model of EMT, the meticulous selection of appropriate cell culture medium formulation is essential to achieve reliable scientific results.

Antiproliferative assay of suma or Brazilian ginseng (Hebanthe eriantha) methanolic extract on HCT116 and 4T1 cancer cell lines, in vitro toxicity on Artemia salina larvae, and antibacterial activity.

Hebanthe eriantha is a medicinal plant used in folk medicine and a subject of commercial interest. The cytotoxicity effects from H. eriantha root extracts on cancerous and normal cells were assessed by the MTT method, and in vitro toxicity was evaluated on Artemia salina. The inhibition of the proliferation of bacteria and MIC values were examined by the disc diffusion and the broth microdilution method, respectively. Human colon cancer HCT116 and mouse breast tumour model 4T1 cells treated with methanolic extract showed a significant decrease in viability of cells with IC50: 272.6 and 88.5 µg/mL at 72h, respectively. The methanolic extract of H. eriantha showed moderate toxicity against A. salina (LC50: 589.4 µg/mL). In antimicrobial activity, the methanolic extract showed the highest inhibitory function against S. aureus and P. vulgaris (17.5 and 16 mm) with MICs of 500 µg/mL. The results confirmed the potential of plant roots as cytotoxic agents.

Increased serum prolactin level may indicate more migraine attack frequency.

OBJECTIVES: Migraine is a common, multifactorial disorder. The exact pathomechanism of migraine remains unclear. Studies have revealed changes in serum prolactin (PRL) levels in relation to migraine, although the results have been inconsistent. The present case-control study assessed the serum level of prolactin in migraine patients. MATERIALS AND METHODS: In this case-control study, participants were divided into chronic migraine (CM; n = 39), episodic migraine in ictal (during an attack), and interictal (between attacks) phases (n = 63, n = 37, respectively) along with 30 age- and sex-matched headache-free controls. After obtaining demographic, anthropometric data, and headache characteristics, blood samples were gathered and analyzed to evaluate the serum levels of prolactin (ng/mL). RESULTS: A significant difference was observed between the control, CM, and ictal EM, and interictal EM groups. The mean ± SD serum prolactin levels of the chronic migraineurs (1.82 ± 0.94) and those with ictal EM (1.93 ± 1.70) were comparable and were significantly higher than for interictal EM patients (0.82 ± 0.46) and the headache-free control subjects (0.49 ± 0.15; p < .001). Although the mean serum concentration of prolactin for the interictal EM group tended to be higher than for control individuals, this difference was not statistically significant. The Spearman's correlation test also showed significant correlations between the serum prolactin levels and the number of headaches days among migraineurs. CONCLUSION: The findings suggest that there might be an association between increased prolactin concentrations and migraine headache induction and progression. Further detailed and well-designed studies are needed to confirm the importance of serum prolactin levels in the pathogenesis of migraine headaches.

In vitro elicitation and detection of apigenin, catalpol and gallic acid in hairy root culture of Plantago major L. and assessment of cytotoxicity and anti-bacterial activity of its methanolic extract.

The aim of this study was to establish the hairy root (HR) culture of Plantago major to evaluate the accumulation of apigenin, catalpol and gallic acid after elicitation and investigate the biological activity of its methanolic extraction. The highest transformation frequency was obtained by Agrobacterium rhizogenes strain A4, 0.5 mg/L 6-Benzylaminopurine in pre-cultivation medium, 150 µM acetosyringone in co-cultivation medium (1/2 MS), and immersion method for inoculation of leaf explants. The production of apigenin, catalpol and gallic acid compounds were significantly affected by treatment of 1.18 mM AgNO3 at 24 h which yielded 4.30, 8.24 and 2.89-fold increase, respectively. The assessment of anti-bacterial activity showed that the methanolic extracts of the HRs elicited with 1.18 mM AgNO3 were significantly active against Proteus vulgaris (PTCC 1182) (MIC = 25 mg/mL and MBC = 25 mg/mL). Furthermore, the MTT assay revealed that the methanolic extracts of the HRs were cytotoxic on the SW-480 cell (IC50=337.56 ± 1.82 µg/mL).

Clinical characteristics of 365 hospitalized COVID-19 patients with neurological symptoms: an observational study.

OBJECTIVE: Since the beginning of the COVID-19 pandemic, a number of COVID-related neurological manifestations have been reported. We aimed to categorize the features of hospitalized COVID-19 patients who experienced neurological symptoms. METHODS: In this descriptive, cross-sectional study, we enrolled all patients hospitalized with COVID-19 who experienced neurological symptoms in two hospitals in Tehran. Diagnosis of COVID-19 was established by PCR tests or computed tomography of the chest combined with COVID-19 clinical findings. The clinical characteristics, laboratory data, and imaging findings from 365 patients were analyzed. RESULTS: The average patient age was 59.2 ± 16.7 years and included 213 males and 152 females. The most prevalent neurological symptoms were headache (56.2%), impaired consciousness (55%), and dizziness (20.5%). During hospitalization, most of the patients did not require mechanical ventilation (81.9%). The percentage of patients with end-organ damage was 9% and mortality was 15%. Regression analysis on the neurological symptoms indicated that the mortality rate of patients with headaches was 84% lower than for the other neurological symptoms. Hyperglycemia was significantly related with end-organ damage and mortality (p = 0.029, p = 0.08, respectively). New vascular lesions were evident on brain MRIs of 9 patients and brain CTs of 16 patients. CONCLUSION: Among the neurological symptoms of patients with COVID-19, headache appeared to indicate a protective factor against development of end-organ damage as well as mortality.

The influence of anxiety and depression on headache in adolescent migraineurs: a case-control study.

BACKGROUND: Migraine is commonly linked to a range of psychiatric comorbidities, primarily anxiety and depression. The present study compared the frequency of anxiety and depression disorders in migraine and non-migraine adolescents and evaluated the relation of depression and anxiety to migraine characteristics. METHODS: In this case-control study, 234 adolescents (112 migraineurs and 122 non-migraine adolescents) aged 13-18 years were evaluated. A headache questionnaire as well as the Beck Anxiety Inventory and Children's Depression Inventory were completed to investigate the prevalence of headache, anxiety, and depression. RESULTS: The average age of participants was 15.77±2 years in the case group and 15.39±1.79 years in the control group. We found significantly higher levels of mild, moderate, and severe anxiety (38.4%, 23.2%and 23.2% respectively) in the migraine group compared to the control group (24.2%, 5.8%and 10.0% respectively) (p<0.001). Significantly more children in the control group (29.6%) than in the case group (10.1%) had a non-depressive CDI score (p=0.005). There was a significant difference between patients with moderate and severe anxiety in terms of attack frequency and duration. Depressed migraineurs recorded higher attack severities and frequencies than migraineurs without depression. CONCLUSION: Anxiety and depression are common in adolescents with migraine and can be associated with more burdensome attacks.

Evaluation of headache associated with personal protective equipment during COVID-19.

BACKGROUND: The COVID-19 pandemic has created new conditions for medical staff, forcing them to use personal protective equipment (PPE) for an extended duration of time. Headache is a commonly associated side effect of the use of such equipment among healthcare workers. METHOD: In this cross-sectional study, 243 frontline healthcare workers at four referral hospitals for COVID-19 were evaluated for the occurrence of headache following the use of PPE and its relationship with blood gas parameters was assessed. RESULTS: The average age of participants was 36 ± 8 years. Of these, 75% were women. The prevalence of headache after the use of masks was 72.4%, with the N95 mask being the most commonly reported cause of headache (41%). Among patients, 25.1% developed external pressure, 22.2% migraine, and 15.2% tension-type headaches. Headache was more common in the female gender. Apart from gender, only increased heart rate was significantly associated with headache due to mask use (p = .03 and .00, respectively). The mean heart rate was 97.7 ± 13.68 in participants with headache compared to 65.8 ± 35.63 in those without headache. No significant relationship was found between headache and venous blood gas parameters, including oxygen and carbon dioxide partial pressure. CONCLUSION: Headache due to PPE is common and can decrease the efficiency of hospital staff performance. Hence, it is necessary to consider this issue among health center personnel and provide modalities to reduce the risk of headache.

The correlation between serum selenium, zinc, and COVID-19 severity: an observational study.

BACKGROUND: Without an adequate immune response, SARS-CoV2 virus can simply spread throughout the body of the host. Two of the well-known immunonutrients are selenium (Se) and zinc (Zn). Se and Zn deficiency might lead to inflammation, oxidative stress, and viral entry into the cells by decreasing ACE-2 expression; three factors that are proposed to be involved in COVID-19 pathogenesis. Thus, in the current study we aimed at evaluating the correlation between serum Se and Zn status and COVID-19 severity. METHODS: Eighty-four COVID-19 patients were enrolled in this observational study. Patients were diagnosed based on an infectious disease specialist diagnosis, using WHO interim guidance and the recommendations of the Iranian National Committee of Covid-19. The patients with acute respiratory tract infection symptoms were checked for compatibility of chest computed tomography (CT) scan results with that of Covid-19 and Real-time polymerase chain reaction (RT-PCR) for corona virus infection. The severity of Covid-19 was categorized into three groups (mild, moderate, and severe) using CDC criteria. Serum Zn and Se level of all subjects was measured. The severity of the disease was determined only once at the onset of disease. RESULTS: According to the results of linear regression test, there was a significant association between Zn and Se level and COVID-19 severity (ß = - 0.28, P-value = 0.01 for Se; ß = - 0.26, P-value = 0.02). However the significance disappeared after adjusting for confounding factors. Spearman correlation analysis showed a significant negative association between serum Zn, Se and CRP level (r = - 0.35, P-value = 0.001 for Se; r = - 0.41, P-value < 0.001 for Zn). CONCLUSION: Results suggest that increasing levels of Se and Zn were accompanied by a decrease in serum CRP level. However, the significant association between Se, Zn, and disease severity was lost after adjusting for confounding factors.

Recent advances in targeting malaria with nanotechnology-based drug carriers.

Malaria, as one of the most common human infectious diseases, remains the greatest global health concern, since approximately 3.5 billion people around the world, especially those in subtropical areas, are at the risk of being infected by malaria. Due to the emergence and spread of drug resistance to the current antimalarials, malaria-related mortality and incidence rates have recently increased. To overcome the aforementioned obstacles, nano-vehicles based on biodegradable, natural, and non-toxic polymers have been developed. Accordingly, these systems are considered as a potential drug vehicle, which due to their unique properties such as the excellent safety profile, good biocompatibility, tunable structure, diversity, and the presence of functional groups within the polymer structure, could facilitate covalent attachment of targeting moieties and antimalarials to the polymeric nano-vehicles. In this review, we highlighted some recent developments of liposomes as unique nanoscale drug delivery vehicles and several polymeric nanovehicles, including hydrogels, dendrimers, self-assembled micelles, and polymer-drug conjugates for the effective delivery of antimalarials.

Nanotechnology against the novel coronavirus (severe acute respiratory syndrome coronavirus 2): diagnosis, treatment, therapy and future perspectives.

COVID-19, as an emerging infectious disease, has caused significant mortality and morbidity along with socioeconomic impact. No effective treatment or vaccine has been approved yet for this pandemic disease. Cutting-edge tools, especially nanotechnology, should be strongly considered to tackle this virus. This review aims to propose several strategies to design and fabricate effective diagnostic and therapeutic agents against COVID-19 by the aid of nanotechnology. Polymeric, inorganic self-assembling materials and peptide-based nanoparticles are promising tools for battling COVID-19 as well as its rapid diagnosis. This review summarizes all of the exciting advances nanomaterials are making toward COVID-19 prevention, diagnosis and therapy.

The effects of vitamin D3 supplementation on TGF-ß and IL-17 serum levels in migraineurs: post hoc analysis of a randomized clinical trial.

BACKGROUND: Although the exact mechanism involved in migraine pathogenesis remained uncertain, and different researches have been developed to address the role of neuroinflammation and immune dysfunction. Therefore, considering the immune protective functions of vitamin D3, we aimed to investigate the effects of daily administration of 2000 IU D3 supplements on serum status of immune markers in migraine patients. METHODS AND MATERIALS: Eighty episodic migraineurs who randomly assigned into two equal groups to receive either vitamin D3 2000 IU/d or placebo for 12-week were enrolled in this placebo-controlled double-blind trial included. Serum concentrations of transforming growth factor-beta (TGF-ß) and interleukin (IL)-17 were evaluated at baseline and after the trial via the ELISA method. RESULTS: Applying ANCOVA adjusted for baseline levels and confounding variables, it was found that the serum level of TGF-ß was significantly higher in vitamin D group (adjusted mean:1665.50 ng/L) than the placebo group (1361.90 ng/L) after the experiment (P-value = 0.012); on the other hand, vitamin D prevented the increment in IL-17 serum level in the intervention group after the trial (adjusted mean:37.84 ng/L) comparing to the controls (adjusted mean:70.09 ng/L; P-value = 0.039). The Pearson correlation analysis revealed a significant positive correlation between changes in serum 25-hydroxy-vitamin D (25(OH)D) and TGF-ß (r = - 0.306, P-value = 0.008). In contrast, no significant correlations were noted between serum 25(OH) D and IL-17 changes throughout the study. CONCLUSION: Based on the results of this study, it was revealed that 12-week vitamin D3 supplementation (2000 IU/day) could enhance the Th17/Treg related cytokines balance in episodic migraineurs. Although these findings are promising, it is needed to be extended. TRIAL REGISTRATION: The trial is registered in the Iranian registry of clinical trials (IRCT) at 11 July 2018, with IRCT code: IRCT20151128025267N6 ( https://www.irct.ir/trial/31246 ).

Anti-Proliferative Properties, Biocompatibility, and Chemical Composition of Different Extracts of Plantago major Medicinal Plant.

Background: To study the anticancer activity of Plantago major, we assessed the effect of ethanolic, methanolic and acetonic extracts of this plant on HCT-116, SW-480, and HEK-293 cell lines as control. Methods: The cytotoxic activity, biocompatibility, and toxicity were evaluated by MTT assay, hemolysis, and Artemia salina-LD50 (on mice) tests, respectively. The analysis of the extracts was performed by GC-MS analysis. Results: The results showed that all the extracts had the most antiproliferative properties on the HCT-116 cell line. The P. major root extract was more effective than the aerial parts, and IC50 values for ethanolic, methanolic and acetonic root extracts were 405.59, 470.16, and 82.26 µg/mL, respectively on HCT-116 cell line at 72 h. Hemolysis degree of the ethanolic extract of aerial and root parts were approximately 1% at 400 µg/mL.. Using the ethanolic extracts, the Artemia survived every concentration, and no toxicity was observed. One week after the oral administration of different parts of P. major extracts, none of the mice died, even those were administered 2000 mg/kg. The results of GC/MS analysis showed that P. major extracts contain potential anticancer compounds, such as stearic acid (8.61%) in aerial parts of methanolic extract and 1,2- Benzenedicarboxylic acid, mono(2-ethylhexyl)ester (88.07% and 40.63%) in aerial and root parts of acetonic extract of P. major. Conclusion: Our findings suggest that the P. major is a source of potential compounds with antiproliferative properties.

Establishment and elicitation of transgenic root culture of Plantago lanceolata and evaluation of its anti-bacterial and cytotoxicity activity.

Hairy root induction in Plantago lanceolata was optimized to take advantage of transformed root cultures. The highest frequency of transformation was achieved using leaf explant, A4 strain, pre-cultivation of explant, 150 µM Acetosyringone, 5 min inoculation, half-strength Murashige and Skoog basal medium as co-cultivation, and half-strength Gamborg's basal medium as a selective medium with 3% sucrose. Among the studied compound encompassing gallic acid, catalpol and apigenin, only the production of gallic acid in hairy roots was affected by 20 mg L-1 AgNO3 and 100 mg L-1 chitosan at 24 hr which yielded 7.63, 4.76-fold increase in its content, respectively. The methanolic extracts of hairy roots elicited by 20 mg L-1 AgNO3 exhibited anti-bacterial activity (MIC and MBC = 25 mg mL-1) against Klebsiella pneumoniae, Proteus vulgaris and Salmonella typhi and anti-bacterial potential of non-elicited hairy roots of P. lanceolata (MIC = 25 mg mL-1 and MBC = 35 mg mL-1) were more active against Klebsiella pneumoniae and P. vulgaris than other bacteria. The methanolic extracts of the P. lanceolata hairy roots demonstrated significant cytotoxic activity on colorectal carcinoma cell line (SW-480) with IC50 = 250.65 ± 6.8 µg mL-1 in comparison to human embryonic kidney (HEK-293) with IC50 = 5263.65 ± 4.6 µg mL-1. Plantago lanceolata hairy roots showed important biological activity explaining its role in traditional medicine.

Estimating quality of life in a headache referral population based on Migraine disability assessment scale and headache impact test.

Background: Headache is among the most common disabling neurologic disorders. We measured quality of life in chronic migraine (CM) and episodic migraine (EM), stratified by medication overuse headache (MOH) and presence of aura. Methods: In this observational study, conducted from January 2016 to December 2018, adult patients referred to the tertiary headache clinic of Sina Hospital in Tehran, Iran, who met International Classification of Headache Disorders, 3rd Edition-beta (ICHD-3 ß) criteria for migraine were classified to EM and CM subtyped based on presence of aura and MOH. Validated Farsi versions of Migraine Disability Assessment Scale (MIDAS) and 6-item Headache Impact Test (HIT-6) questionnaires were used. Results: A total of 2454 patients (1907 women) were enrolled from which 1261 (51.4%) patients had EM and 1193 (48.6%) had CM, while 908 subjects (37.0%) had MOH, of whom 890 (98.0%) had CM. Median scores of MIDAS and HIT-6 were significantly higher in patients with CM compared to EM sufferers. Chronic migraineurs with MOH had a significantly higher median score of MIDAS and HIT-6 compared to patients with non-MOH CM. Also, there was a moderate positive correlation between MIDAS (disability) and HIT-6 scores (impact on patients' life) and a moderate correlation between HIT-6 and pain severity. Conclusion: The results of this study confirm that CM and MOH are associated with a higher headache-related disability and impact on life compared to EM. Therefore, treatment goals in prevention of MOH and migraine transformation warrant higher quality of life in patients with migraine.

Anti-carcinogenic and anti-angiogenic properties of the extracts of Acorus calamus on gastric cancer cells.

OBJECTIVE: Acorus calamus (A. calamus) has been used as a medicinal plant in Asia for its effects on digestive system for the last 2000 years. To investigate the anti-cancer activity of rhizome of A. calamus, the ethanolic and methanolic extracts and essential oil of the rhizome were prepared and their effects were assessed on human gastric cancer cell line (AGS). MATERIALS AND METHODS: The viability of cells which were treated with the extracts and the essential oil was assessed by MTT assay. To evaluate the anti-angiogenic property of the extracts, in vitro tube formation assay was done. Cell cycle distribution and the expression of Oct4 and Nucleostemin, after treatments, were checked by flowcytometry and quantitative RT-PCR, respectively. Furthermore, analysis of essential oil from A.calamus was done by GC-MS. RESULTS: Our results showed that the growth of AGS cells was inhibited by the extracts and essential oil and the extracts inhibited the angiogenesis in HUVEC cells. Our data revealed that the extracts and essential oil of A. calamus caused G1 arrest in AGS cells and downregulation of Oct4 and NS after treatment. By GC-MS analysis, we found new compounds such as epiprezizaene, valencene and isocyclocitral in essential oil of A. CONCLUSION: All together, our results showed that the extracts of A. calamus have anti-proliferative and anti-angiogenic effects on cancer cells.

Parapharengeal unicentric Castleman disease with nephrotic syndrome.

Castleman disease (CD) is a rare lymphoproliferative disorder, first described in 1956. This case report describes a 27-year-old man with hyaline vascular unicentric CD, first presented with edema and hypertension. On initial evaluation for edema, 24-hour urine collection revealed 8200 mg/24 h protein excretion. Pathologic examination of the kidney specimen showed diffuse mesangial lesions with segmental subepithelial deposition. On follow-up for nephrotic syndrome, the patient experienced a feeling of a mass in his pharynx and deterioration of previous snoring, documented by neck magnetic resonance imaging. Pathology report of the excisional biopsy showed CD. Treatment with corticosteroids and partial excision can be considered as an alternative to surgery for unresectable unicentric CD. The 5-year follow-up showed that this strategy could lead to remission.