FTIR microspectroscopy reveals chemical changes in mice fetus following Phenobarbital administration

Phenobarbital is a phenobarbiturate used as a sedative, anticonvulsant or hypnotic with the doses prescribed and can cause teratogenic effects. The goal of this study was to examine an alternative method for the recognition of the mechanism or the bimolecular potential changes in mice fetus caused by Phenobarbital using FTIR micro spectroscopy. The mice were injected with Phenobarbital (120 mg/Kg) on gestation day 9. Fetuses were dissected on day 15 of gestation and morphological and histological studies on the fetus were carried out. Sections (10 μm) of normal and Phenobarbital treated fetus brains and livers were used for FTIR measurement in the wave number region of 400- 4000 cm. The results were shown by 2 derivatization of spectra and also subtracting from control spectra. In liver, the intensity at 1054 cm, 1155 cm, 1353 cm, 1453cm,1645 cm, 1622 cm, 2944 cm, 2913 cm and 2845 cm were shifted and increased. In the brain, the intensity at 879 cm, 911 cm, 955 cm, 1223 cm, 1256 cm, 1304 cm, 1360 cm, 1453 cm, 1529 cm, 1636 cm, 2845 cm, 2915 cm and 2950 cm were increased and shifted. The most important changes of the fetus brain tissue are on the β structure of proteins due to the amide I bands at 1636 cm, while extensive effects on the DNA structure were obvious for the Phenobarbital treated liver tissues. As a conclusion, FTIR spectroscopy might well be assumed as a potentially powerful teratogenic measurement instrument with a unique ability to identify the modified bimolecular structures. © 2015 by School of Pharmacy Shaheed Beheshti University of Medical Sciences and Health Services

In Vitro and in Vivo Enhancement of Antitumoral Activity of Liposomal Antisense Oligonucleotides by Cineole as a Chemical Penetration Enhancer

Cellular uptake and cytoplasmic release of liposomal antisense oligonucleotides (AsODNs), which can act as rate-limiting steps, are still remained to be completely optimized. Here, the possibility of enhancing such processes at cellular and animal levels by cineole, as a penetration enhancer, was investigated. A cationic nanoliposome containing an AsODN against PKC-α and a cineole-containing nanoliposome were prepared and characterized. The effect of nanoliposomal cineole on sequence-specific cytotoxicity of nanoliposomal AsODN against A549, was studied in vitro (MTT, flow cytometry, fluorescence microscopy, and real time PCR) and in vivo (xenograft lung tumor in nude mice) using different concentrations and treatment times. Results showed specific cytotoxicity of nanoliposomal AsODN was increased significantly from 11 to 25 when A549 cells were exposed to 10 μg/mL cineole for 1 or 4 hours. This inhibitory effect was further increased to about 40 when the concentration was increased to 40 μg/mL for 1 hour. In animal studies, cineole significantly decreased the tumor volume (about 75) and increased its doubling time from 13 days to 31 days. A linear relationship exists between cineole concentration and its enhancement effects. Finally it was concluded that cineole, and possibly other membrane fluidizers, can improve nanoliposomal gene therapy at cellular and animal levels. © 2015 Hamid Reza Moghimi et al

Human lung carcinoma reaction against metabolic serum deficiency stress

Cancer treatment is still of the greatest challenges that health care providers and patients are facing. One of the unsolved problems in cancer treatment is cells� reaction to metabolic stress caused by harsh nutritional conditions around tumor. In order to be able to treat this disease properly, it is important to understand the true nature of the disease. In fact, the cells inside the central part of the tumor lack sufficient access to blood vessels, nutrients, and growth signals. After tumor shrinkage, the cells are exposed to favorable environmental conditions and might regrow and cause tumor recurrence. The main purpose of this study was to investigate the effect of serum starvation, as a type of metabolic stress, on human lung cancer cell line, A549. These cells were treated with 10 (control), 0.5 and 0.25 serum for 1 to 5 days. At 24 h intervals, the cells were released with 10 serum supplemented media. Starved or released cells were studied for their cycle and morphology. The results showed that the cells were actually arrested at G1 phase and following exposure to optimal conditions, the cells could be back to their cycle again. Furthermore, sub-G1 apoptotic cells population was not increased within the starvation period, while control cells had significant increase in sub-G1 cells. Morphological studies also showed that starved cells could make denser colonies while control cells were entering death phase. These observations provide some evidence for the generation of some effective resistance phenomena in cancer cells against harsh metabolic conditions © 2016 by School of Pharmacy

The role of Olea Europaea L. Fruit on A2780, A172 and HFFF2 proliferation

Olea europaea L. commonly known as olive has been traditionally used for the prevention and treatment of many diseases since ancient times. Olive has been reported to possess a broad spectrum of pharmacological properties. In the present study, we investigated the activity of aqueous extract of Olea europaea L. fruit at various concentrations on A2780, A172, and HFFF2 cell lines proliferation by MTT assay. Aqueous extract of olive significantly increased cell proliferation in a dose dependent manner in the cell lines. It has been previously reported that olive has chemoproventive and anti-tumor effects. These disagreements can be explained by differences in cell line properties, type of olive and different solvents in the extracts. However, further investigation is needed to clarify the exact role of olive in cell proliferation and cancer. In this study fruit extract of Olea europaea L. showed more activatory effects on A2780 cell line in comparison with A172 and HFFF2. These differences in the activatory effects may be related to the activation of different signaling pathways in different cell lines. © 2016, Iranian Association of Pharmaceutical Scientists. All rights reserved

FTIR determination of miconazole effects on mice fetus brain tissue

Miconazole is an imidazole antifungal agent, commonly applied topically to the skin or mucous membranes. The aim of this study was to examine the alternative method for gaining mechanism or the bimolecular changes caused by the possible teratogenic effects of Miconazole on mice fetus brain tissue using FTIR-Microspectroscopy. The mice were injected with Miconazole (60 mg/Kg) on gestation day 9. Fetuses were dissected on day 15 of gestation and morphological and histological studies on the fetus were carried out. Sections (10 μm) of control and Miconazole treated fetus brain tissue were used for FTIR measurement in the mid- infrared region. The results were shown by spectra 2nd derivative and also subtracting from control spectra. A lower intensity in the lipid (2800–3000 cm-1) and amid I (1600–1800 cm-1) regions of Miconazole treated mice fetus brain tissue was observed compared to the control mice fetus brain tissue. No major spectral shifting was observed at amide I band, amide II band and nucleic acid regions. As a conclusion, FTIR-Microspectroscopy can be a useful tool for teratogenic measurement with a unique ability to identify the modified bimolecular structures in mice fetus tissues. © 2014, Iranian Association of Pharmaceutical Scientists. All rights reserved

Male infertility in iranian traditional medicine, causes, treatment and compares it with modern medicine

Introduction: One of the major problems of medical science is infertility and reduced fertility. At least one of six couples is infertile. Male infertility is considered in 40 of infertility cases. Prevalence and importance of infertility and the attention of traditional medicine scholars to the causes of infertility and treatment of this problem which its main part is recipes can be helpful, because nutrition has many effects on human health. This study was performed with aim to evaluate the causes of infertility and recipes mentioned in traditional medicine books to prevent and treat male infertility beside modern medicine. Methods: In this systematic review study, the main textbooks of traditional medicine such as Ghanoon in the section of male fertility disorders were evaluated according to the key words of infertility and nutrition, and the causes of male infertility were studied. Then, the causes of male infertility and its treatment in the books and articles of modern medicine were studied and the results were briefly compared. Results: Iranian traditional physicians have described several causes for infertility and have expressed different treatments depending on the cause. Among these, the role of nutrition is highlighted. The causes of this disorder in some cases is weakness, in some cases is weakness in main organs such as brain, heart, liver and testicles, and in some cases is the main organs's mizaj. Simple food regimes can be used for treatment of many cases, therefore, in this study, treatment and nutritional measures were obtained based on the cause. Conclusion: The studies showed that viewpoints are similar in Traditional Medicine and Modern Medicine); in both viewpoints, the causes of infertility can be related to the reproductive system (testicles, tract, and penis), sometimes brain, liver and kidney dysfunction, and in some cases, sexual dysfunction may be involved in infertility. These similarities show the richness of Iranian medicine and need for attention to its therapy methods. According to the results, as the first step of treatment, the list of foods presented in this study can be used. This list can be recommended to the patients in medical centers. © 2016, Mashhad University of Medical Sciences. All rights reserved

Terpene-loaded liposomes and isopropyl myristate as chemical permeation enhancers toward liposomal gene delivery in lung cancer cells; a comparative study

Gene therapy is in its development stage as a novel method for cancer treatment. Liposomes look promising as gene delivery vectors; however, investigations have shown that these vesicles are not doing well in some cases. It was decided here to investigate the possibility of augmentation of liposomal gene delivery by chemical penetration enhancers.Cationic liposome containing antisense oligonucleotide (AsODN) against lung cancer was prepared by ethanol injection method. Liposomal cineole and limonene (as enhancers) were prepared by film hydration method. Isopropyl myristate (IPM) was also investigated as penetration enhancer. Liposomes were evaluated for their size, zeta potential and encapsulation efficiency. Cancer cells (A549) were pretreated with liposomal terpenes prior to treatment with liposomal antisense or scrambled oligonucleotide. Cell viability was evaluated by MTT assay.Oligonucleotide-containing liposome showed particle size of about115 nm and zeta potential of 0.6 mV. Liposomal cineole significantly (P<0.05) increased specific activity of liposomal antisense but limonene didn�t show such an effect. IPM increased both specific and non-specific cytotoxicity of oligonucleotide.These results show that penetration enhancers (such as cineole) may be used for improving liposomal gene delivery and to reduce non-specific toxicity. Concentration and chemical nature of enhancer has prominent effect in their efficacy. © 2016 by School of Pharmacy

Evaluation of mutagenicity of mebudipine, a new calcium channel blocker

Mebudipine is a new dihydropyridine calcium channel blocker, synthesized in our laboratory, for treatment of hypertension. It has shown a better efficacy than other drugs in this group. For assessing the risks of this drug, certain safety tests in the preclinical stage have been performed. In this study mutagenic effect of mebudipine was evaluated using Ames assay that could assess the mutagenicity of drugs and their metabolites using liver enzymes (S-9 mix). This procedure is approved as a predictive test, with a high predictive value. Salmonella TA102 (Ames assay) was used with and without S-9 in this study. For preparing S-9 mix, rat liver enzymes induced by phenobarbital were separated in KCl 0.154 M (0.154 M), as the solvent. Mebudipine was dissolved in polyethylenglycol 400. Mutagenicity test was performed in 6 doses from 39 μg to 1250 μg per every plate, in the presence and absence of the S-9 mix. The positive control sodium azide was dissolved in a dose of 5 μg/plate dissolved in polyethylenglycol 400 and negative control was polyethylenglycol 400 with no added agent. The colony counts of all doses in plates with S-9 were between 200-400 and in plates without S9 was between100-300. The colony counts in both states (with and without S-9) of all doses were in the range suggested by Ames assay for the safe drugs and were different from the positive control groups and equal to the negative controls. Mebudipine and its metabolites were not found to be mutagen on Salmonella TA102, based on Ames assay. Copyright © 2010 by School of Pharmacy Shaheed Beheshti University of Medical Sciences and Health Services

Teratogenic study of phenobarbital and levamisole on mouse fetus liver tissue using biospectroscopy

Biospectroscopic investigations have attracted attention of both the clinicians and basic sciences researchers in recent years. Scientists are discovering new areas for FTIR biospectroscopy applications in medicine. The aim of this study was to measure the possibility of FTIR-MSP application for the recognition and detection of fetus abnormalities after exposure of pregnant mouse to phenobarbital (PB) and levamisole (LEV) alone or in combination. PB is one of the most widely used antiepileptic drugs (AEDs), with sedative and hypnotic effects. When used by pregnant women, it is known to be a teratogenic agent. LEV is an antihelminthic drug with some applications in immune-deficiency as well as colon cancer therapy. Four groups of ten pregnant mice were selected for the experiments as follows: one control group received only standard diet, one group was injected with 120 mg/kg of BP, one group was injected with 10 mg/kg of LEV, and the last group was treated simultaneously with both BP and LEV at the above mentioned doses. Drugs administration was performed on gestation day 9 and fetuses were dissected on pregnancy day 15. Each dissected fetus was fixed, dehydrated and embedded in paraffin. Sections of liver (10 μm) were prepared from control and treated groups by microtome and deparaffinized with xylene. The spectra were taken by FTIR-MSP in the region of 4000�400 cm�1. All the spectra were normalized based on amide II band (1545 cm�1) after baseline correction of the entire spectrum, followed by classification using PCA, ANN and SVM. Both morphological and spectral changes were shown in the treated fetuses as compared to the fetuses in the control group. While cleft palate and C-R elongation were seen in PB injected fetuses, developmental retardation was mostly seen in the LEV injected group. Biospectroscopy revealed that both drugs mainly affected the cellular lipids and proteins, with LEV causing more changes in amide I and lipid regions than PB. Application of PCA, ANN and SVM methods were able to successfully classify these FTIR spectroscopic data and discriminate between control and treated groups of fetuses, making it a new potential tool for drugs teratogenic investigations. © 2016 Elsevier B.V

Anti-hyperglycemic effect of aqueous extract of Juglans regia L. leaf (walnut leaf) on type 2 diabetic patients: A randomized controlled trial

Objective: The Juglans regia leaf (walnut leaf) has been traditionally used to treat diabetes in Iran. The aim of this study was to evaluate the effect of aqueous extract of Juglans regia leaf on blood glucose in type 2 diabetic patients. Methods: Thirty-seven type 2 diabetic patients aged 35�70 years with fasting blood glucose levels between 130 mg/dl (7.2 mmol/L) and 250 mg/dl (13.9 mmol/L) and HbA1c more than 7 despite using conventional oral hypoglycemic drugs received the Juglans regia capsule (250 mg/TDS) or toast powder capsule (placebo) randomly for 3 months. Fasting blood glucose (FPG), postprandial blood glucose (PPG), and glycosylated hemoglobin (HbA1c) were assessed as primary outcome measures. Lipid profile and hepatic and renal function were assessed as secondary outcome measures. Nonparametric statistical tests were employed to compare the results with baseline in both groups using SPSS version 17. Results: Nineteen patients in the Juglans regia group and 18 in the control group with a mean (±SD) age 55.8 (±7) completed the study. The FPG (p = 0.02), PPG (p = 0.008) and HbA1c (p = 0.003) levels decreased significantly in the Juglans regia group after 3 months compared to baseline without any side effects. The changes of the variables were not significant in the control group. Conclusions: This study suggests that the aqueous extract of J. regia leaf had a lowering effect of FPG, PPG and HbA1c in type 2 diabetic patients. Conducting further clinical studies is suggested with larger sample sizes to make precise conclusions about the efficacy and safety of this herb. © 2017 Elsevier Lt