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

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

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