Joanna Depciuch

Application of Raman Spectroscopy and Infrared Spectroscopy in the Identification of Breast Cancer

Raman spectroscopy and infrared (IR) spectroscopy are both techniques that allow for the investigation of vibrating chemical particles. These techniques provide information not only about chemical particles through the identification of functional groups and spectral analysis of so-called “fingerprints”, these methods allow for the qualitative and quantitative analyses of chemical substances in the sample. Both of these spectral techniques are frequently being used in biology and medicine in diagnosing illnesses and monitoring methods of therapy. The type of breast cancer found in woman is often a malignant tumor, causing 1.38 million new cases of breast cancer and 458 000 deaths in the world in 2013. The most important risk factors for breast cancer development are: sex, age, family history, specific benign breast conditions in the breast, ionizing radiation, and lifestyle. The main purpose of breast cancer screening tests is to establish early diagnostics and to apply proper treatment. Diagnoses of breast cancer are based on: (1) physical techniques (e.g., ultrasonography, mammography, elastography, magnetic resonance, positron emission tomography [PET]); (2) histopathological techniques; (3) biological techniques; and (4) optical techniques (e.g., photo acoustic imaging, fluorescence tomography). However, none of these techniques provides unique or especially revealing answers. The aim of our study is comparative spectroscopic measurements on patients with the following: normal non-cancerous breast tissue; breast cancer tissues before chemotherapy; breast cancer tissues after chemotherapy; and normal breast tissues received around the cancerous breast region. Spectra collected from breast cancer patients shows changes in amounts of carotenoids and fats. We also observed changes in carbohydrate and protein levels (e.g., lack of amino acids, changes in the concentration of amino acids, structural changes) in comparison with normal breast tissues. This fact verifies that Raman spectroscopy and IR spectroscopy are very useful diagnostic tools that will shed new light in understanding the etiology of breast cancer.

FPA-FTIR Microspectroscopy for Monitoring Chemotherapy Efficacy in Triple-Negative Breast Cancer

Triple-negative breast cancer is the most aggressive breast cancer subtype with limited treatment options and a poor prognosis. Approximately 70% of triple-negative breast cancer patients fail to achieve a pathologic complete response (pCR) after chemotherapy due to the lack of targeted therapies for this subtype. We report here the development of a focal-plane-array Fourier transform infrared (FPA-FTIR) microspectroscopic technique combined with principal component analysis (PCA) for monitoring chemotherapy effects in triple-negative breast cancer patients. The PCA results obtained using the FPA-FTIR spectral data collected from the same patients before and after the chemotherapy revealed discriminatory features that were consistent with the pathologic and clinical responses to chemotherapy, indicating the potential of the technique as a monitoring tool for observing chemotherapy efficacy.

The role of zinc deficiency-induced changes in the phospholipid-protein balance of blood serum in animal depression model by Raman, FTIR and UV–vis spectroscopy

Depression is a serious mental illness. To study the mechanisms of diseases and search for new, more effective therapies, animal models are used. Unfortunately, none of the available models does reflect all symptoms of depression. Zinc deficiency is proposed as a new animal model of depression. However, it has not been yet validated in a detailed manner. Recently, spectroscopic techniques are increasingly being used both in clinical and preclinical studies. Here we examined the effect of zinc deficiency and amitryptyline treatment on the phospholipid - protein balance in the blood serum of rats using Raman, Fourier Transform Infra Red (FTIR) and UV-vis technique. Male Sprague Dawley rats were fed with a zinc ample diet (ZnA, 50mg Zn/kg) or a zinc deficient diet (ZnD, 3mg Zn/kg) for 4 weeks. Then amitriptyline administration (AMI, 10mg/kg, i.p.) was started. After injecting the drug for 2-weeks, blood samples were collected and analyzed. It was found that zinc deficiency decreases both the level of phospholipids and proteins and also causes structural changes in their structures. In the ZnD group amitriptyline treatment influenced the protein level and structure. UV-vis spectroscopy combined with the second derivative calculated from the FTIR spectra provided information that the proteins in blood serum of rat fed with a low Zn diet regain their intact structure after amitriptyline medication. Simultaneously, the antidepressant therapy did not have any effect on the level of phospholipids in this group of rats. Additionally, our results show, that amitriptyline administration can change the structure of phospholipids in rats subjected to zinc ample diet. This altered structure of phospholipids was identified as shortening of carbon chains. Our findings indicate that the decreased level of zinc may be the cause of depressive disorders, as it leads to changes in the phospholipid-protein balance necessary for the proper functioning of the body. This study also shows possible new applications of spectroscopic techniques in the diagnosis of affective disorders, and maybe even identifies markers of depressive disorders.

FTIR analysis of molecular composition changes in hazel pollen from unpolluted and urbanized areas

In this study, the effect of urbanization and environmental pollution on qualitative (structural) and quantitative changes of the Corylus avellana (hazel) pollen was investigated using scanning electron microscopy, Fourier Transform Infrared (FTIR) spectroscopy and curve-fitting analysis of amide I profile. The obtained spectroscopic results show significant variations in the fraction of proteins in the hazel pollen, which probably depend on various degrees of anthropopression. Our results suggest that alterations in the chemical composition of pollen, induced by urbanization and air pollutants, may intensify the allergenic potential and may cause the increase in the incidence of allergies in people. Mutations in nucleic acids are accompanied by a number of molecular changes leading to the formation of allergenic proteins. It seems that the type of habitat, where the pollen grew, affects the individual differentiation. Indeed, it was found that in the site exhibiting low pollution, the hazel pollen contain a lower amount of proteins than to the ones from a site with high anthropopression. Hence, FTIR spectroscopy and curve-fitting analysis of amide I profile can be successfully applied as tools for identifying quantitative and qualitative changes of proteins in hazel pollen.Graphical AbstractAnthropogenic factors such as air pollution and urbanization lead to changes in structure and chemical composition of hazel pollen. Fourier Transform Infrared spectroscopy (FTIR) and Gaussian analysis showed structural changes in hazel pollen collected from sites with different absorbance values of individual chemical functional groups and changes in the secondary structure of proteins of the pollen.

Use of FTIR spectroscopy and PCA-LDC analysis to identify cancerous lesions within the human colon.

HIGHLIGHTSFTIR is a sensitive distribution indicator for the main biochemical compounds of healthy and cancerous colon tissue.Cancerogenesis caused changes in spectrum representing the vibration of nucleic acids, protein and lipid functional groups.Post‐chemotherapy colon tissues FTIR spectrum was similar with healthy colon tissues. ABSTRACT Colorectal cancer constitutes 33% of all cancer morbidity, so the research of the new methods for colorectal cancer diagnosis and chemotherapy monitoring is gaining its momentum. Diagnostic instruments are being sought, which enable the detection of single malignant cells based on the analysis of tissue material potentially reusable at further stages of diagnostic management. The most common approach to tissue specimen processing is paraffin‐embedding. Yet, paraffin may cause background noise in spectroscopic measurements with the wavenumber ranging between 900 cm−1 and 3500 cm−1. However, the study by Depciuch et al. (2016) proved that appropriate specimen processing and paraffin‐embedding technique as well as a strict measurement methodology may eliminate paraffin vibrations. As a result, spectroscopic measurements may become a reliable and precise method for the diagnosis and treatment monitoring in patients with colorectal cancer as long as the high standards of specimen processing are maintained. Chemotherapy is the main medical treatment in colorectal cancer. Unfortunately, the absence of tools which enable monitoring its efficacy leads to the partial response or non‐response frequently seen in affected patients. Hence, diagnostic instruments are also being sought capable of monitoring treatment efficacy so as to enable early changes of chemotherapy regimen thus increasing the chance of cure. The paper aims at comparing the results of FTIR (Fourier Transform Infrared) spectroscopy in several types of colon tissue: healthy colon, cancerous colon, post‐chemotherapy colon and healthy surgical margin of colon cancer sample. The obtained FTIR spectra along with the Principal Component Analysis‐Linear Discriminant Analysis (PCA‐LDC) as well as bandwidth analysis of the primary amide region revealed some differences between the spectra of healthy tissues as compared to cancerous tissues (pre‐ or post‐chemotherapy). Apart from confirming that FTIR spectroscopy is a good source of information on the composition of analysed samples, this fact supports its application as a tool to facilitate understanding the pathophysiology of various conditions and to monitor efficacy of chemotherapy in cancer patients.

Qualitative and quantitative changes in phospholipids and proteins investigated by spectroscopic techniques in animal depression model

Depression becomes nowadays a high mortality civilization disease with one of the major causes being chronic stress. Raman, Fourier Transform Infra Red (FTIR) and Ultraviolet-Visible (UV-vis) spectroscopies were used to determine the changes in the quantity and structure of phospholipids and proteins in the blood serum of rats subjected to chronic mild stress, which is a common animal depression model. Moreover, the efficiency of the imipramine treatment was evaluated. It was found that chronic mild stress not only damages the structure of the phospholipids and proteins, but also decreases their level in the blood serum. A 5weeks imipramine treatment did increase slightly the quantity of proteins, leaving the damaged phospholipids unchanged. Structural information from phospholipids and proteins was obtained by UV-vis spectroscopy combined with the second derivative of the FTIR spectra. Indeed, the structure of proteins in blood serum of stressed rats was normalized after imipramine therapy, while the impaired structure of phospholipids remained unaffected. These findings strongly suggest that the depression factor, which is chronic mild stress, may induce permanent (irreversible) damages into the phospholipid structure identified as shortened carbon chains. This study shows a possible new application of spectroscopic techniques in the diagnosis and therapy monitoring of depression.

Verification of the effectiveness of the Fourier transform infrared spectroscopy computational model for colorectal cancer

HighlightsPhysics‐based computational model reflects the efficacy of chemotherapy.Post‐chemotherapy colon tissues FTIR spectrum was similar with healthy one.FTIR is a sensitive distribution indicator for the main biochemical compounds of healthy and cancerous colon tissue.Cancerogenesis caused changes in spectrum representing the vibration of carbohydrate, protein and lipid functional groups. Abstract Colorectal cancer is one of the most common cancers. Its formation is influenced by genetic and environmental factors. Despite the continuous development of diagnostic tools and cancer therapies, there are no methods that allow a real‐time estimation of treatment efficiency. This method can be a vibrational spectroscopy. The resulting infrared spectrum (FTIR) of the tissue gives us information about the chemical composition and the content of the individual components. We have noticed that tumor tissues, healthy and after chemotherapy tissues, have different vibrational spectra. It was also shown that spectra acquired from normal (benign) tissues were similar to those derived from tissues post‐chemotherapy. The similarity was greater, when the effectiveness of chemotherapy, confirmed by medical documentation, was better. Therefore, we decided to use the physical model proposed in our earlier paper to verify its correctness and to show whether a particular type of chemotherapy was effective or not. Comparison of the results obtained from the physical model with patients data have been found as close to the physical condition.

Monitoring breast cancer treatment using a Fourier transform infrared spectroscopy-based computational model

&NA; Breast cancer affects one in four women, therefore, the search for new diagnostic technologies and therapeutic approaches is of critical importance. This involves the development of diagnostic tools to facilitate the detection of cancer cells, which is useful for assessing the efficacy of cancer therapies. One of the major challenges for chemotherapy is the lack of tools to monitor efficacy during the course of treatment. Vibrational spectroscopy appears to be a promising tool for such a purpose, as it yields Fourier transformation infrared (FTIR) spectra which can be used to provide information on the chemical composition of the tissue. Previous research by our group has demonstrated significant differences between the infrared spectra of healthy, cancerous and post‐chemotherapy breast tissue. Furthermore, the results obtained for three extreme patient cases revealed that the infrared spectra of post‐chemotherapy breast tissue closely resembles that of healthy breast tissue when chemotherapy is effective (i.e., a good therapeutic response is achieved), or that of cancerous breast tissue when chemotherapy is ineffective. In the current study, we compared the infrared spectra of healthy, cancerous and post‐chemotherapy breast tissue. Characteristic parameters were designated for the obtained spectra, spreading the function of absorbance using the Kramers–Kronig transformation and the best fit procedure to obtain Lorentz functions, which represent components of the bands. The Lorentz function parameters were used to develop a physics‐based computational model to verify the efficacy of a given chemotherapy protocol in a given case. The results obtained using this model reflected the actual patient data retrieved from medical records (health improvement or no improvement). Therefore, we propose this model as a useful tool for monitoring the efficacy of chemotherapy in patients with breast cancer. HighlightsFTIR is a sensitive distribution indicator for the main biochemical compounds of healthy and cancerous breast tissue.Cancerogenesis caused changes in spectrum representing the vibration of carbohydrate, protein and lipid functional groups.Physics‐based computational model was postulated to determine the efficacy of chemotherapy.Post‐chemotherapy breast tissues FTIR spectrum was similar with healthy breast tissues.

The classification of lung cancers and their degree of malignancy by FTIR, PCA-LDA analysis, and a physics-based computational model

Lung cancer has the highest mortality rate of all malignant tumours. The current effects of cancer treatment, as well as its diagnostics, are unsatisfactory. Therefore it is very important to introduce modern diagnostic tools, which will allow for rapid classification of lung cancers and their degree of malignancy. For this purpose, the authors propose the use of Fourier Transform InfraRed (FTIR) spectroscopy combined with Principal Component Analysis-Linear Discriminant Analysis (PCA-LDA) and a physics-based computational model. The results obtained for lung cancer tissues, adenocarcinoma and squamous cell carcinoma FTIR spectra, show a shift in wavenumbers compared to control tissue FTIR spectra. Furthermore, in the FTIR spectra of adenocarcinoma there are no peaks corresponding to glutamate or phospholipid functional groups. Moreover, in the case of G2 and G3 malignancy of adenocarcinoma lung cancer, the absence of an OH groups peak was noticed. Thus, it seems that FTIR spectroscopy is a valuable tool to classify lung cancer and to determine the degree of its malignancy.

Spectroscopic analysis of normal and neoplastic (WI-FTC) thyroid tissue

Thyroid cancer holds the first place of the malignant tumors of the endocrine system. One of the less common thyroid cancers is follicular thyroid carcinoma (FTC), which is very difficult to diagnose because it gives the same image as adenoma, which is benign. Certainty of the diagnosis is gained only when FTC gives metastases. Therefore, it was decided to compare normal and neoplastic (FTC) thyroid tissues with Fourier Transform Infrared (FTIR) spectroscopy. The obtained FTIR spectra and Principal Component Analysis (PCA) allowed us to conclude that there are differences in the FTIR spectrum between normal tissues and those affected by cancer. In addition, the results indicate that there is a decrease in the number of functional groups that build cellular and tissue structures in tumoral tissues. The shifts of wave numbers corresponding to the protein and lipid function group vibrations, as well as the calculated second derivative of the FTIR spectra showed the structural changes in neoplastic tissues. Moreover, the deconvolution of the amide I massif indicates that in cancerous tissues the prevailing secondary structure is β-sheet structure, while in normal tissues it is α-helix. The obtained results allow us to conclude that infrared spectroscopy, in addition to providing information on the composition of tested samples, can be an excellent diagnostic tool contributing to understanding the FTC substrate.