Molecular spectroscopic markers of DNA damage

Every cell in a living organism is constantly exposed to physical and chemical factors which damage the molecular structure of proteins, lipids, and nucleic acids. Cellular DNA lesions are the most dangerous because the genetic information, critical for the identity and function of each eukaryotic cell, is stored in the DNA. In this review, we describe spectroscopic markers of DNA damage, which can be detected by infrared, Raman, surface-enhanced Raman, and tip-enhanced Raman spectroscopies, using data acquired from DNA solutions and mammalian cells. Various physical and chemical DNA damaging factors are taken into consideration, including ionizing and non-ionizing radiation, chemicals, and chemotherapeutic compounds. All major spectral markers of DNA damage are presented in several tables, to give the reader a possibility of fast identification of the spectral signature related to a particular type of DNA damage

Echo rozwiązuje kliniczną zagadkę — opis przypadku amyloidozy serca

We present the case of a 53-year-old woman with New York Heart Association (NYHA) class III heart failure, despite optimal pharmacological therapy. Her medical history included: myocardial infarction with nonobstructive coronary arteries (MINOCA), heart failure with reduced ejection fraction and quadrantanopia with sensory neuropathy diagnosed as ischaemic stroke. Transthoracic echocardiography showed significant left ventricular hypertrophy with a ‘granular’ myocardial texture, global systolic dysfunction and restrictive mitral filling pattern. Two-dimensional (2D) transthoracic echo strain analysis demonstrated decreased global longitudinal strain with apical sparing, and helped to fit all the pieces of the jigsaw together to reach a consistent clinical diagnosis of cardiac amyloidosis.Przedstawiono opis przypadku 53-letniej kobiety z obniżoną tolerancją wysiłku fizycznego w III klasie czynnościowej według New York Heart Association (NYHA), mimo optymalnej farmakoterapii. W wywiadzie stwierdzono: przebyty zawał serca bez istotnych zmian w nasierdziowych tętnicach wieńcowych (MINOCA), niewydolność serca z obniżoną frakcją wyrzutową lewej komory oraz niedowidzenie kwadrantowe z neuropatią czuciową zdiagnozowane jako udar niedokrwienny mózgu. W echokardiograficznym badaniu przezklatkowym (TTE) uwidoczniono „ziarnistą” strukturę oraz znaczny przerost mięśnia sercowego lewej komory, uogólnioną dysfunkcję skurczową oraz restrykcyjny profil napływu mitralnego. Analiza dwuwymiarowa (2D) TTE wykazała zmniejszenie globalnego odkształcenia podłużnego z lepszą funkcją w obrębie koniuszka. Charakterystyczny obraz echokardiograficzny przyczynił się do złożenia wszystkich elementów klinicznych w diagnozę amyloidozy serca

Affinity of alkylphosphocholines to biological membrane of prostate cancer : studies in natural and model systems

The effectiveness of two alkylphosphocholines (APCs), hexadecylphosphocholine (miltefosine) and erucylphosphocholine to combat prostate cancer has been studied in vitro with artificial cancerous membrane, modelled with the Langmuir monolayer technique, and on cell line (Du-145). Studies performed with the Langmuir method indicate that both the investigated drugs have the affinity to the monolayer mimicking prostate cancer membrane (composed of cholesterol:POPC = 0.428) and the drug-membrane interactions are stronger for erucylphosphocholine as compared to hexadecylphosphocholine. Moreover, both studied drugs were found to fluidize the model membrane, which may lead to apoptosis. Indeed, biological studies confirmed that in Du-145 cell line both investigated alkylphosphocholines cause cell death primarily by apoptosis while necrotic cells constitute only a small percentage of APC-treated cells

Molecular spectroscopic markers of abnormal protein aggregation

Abnormal protein aggregation has been intensively studied for over 40 years and broadly discussed in the literature due to its significant role in neurodegenerative diseases etiology. Structural reorganization and conformational changes of the secondary structure upon the aggregation determine aggregation pathways and cytotoxicity of the aggregates, and therefore, numerous analytical techniques are employed for a deep investigation into the secondary structure of abnormal protein aggregates. Molecular spectroscopies, including Raman and infrared ones, are routinely applied in such studies. Recently, the nanoscale spatial resolution of tip-enhanced Raman and infrared nanospectroscopies, as well as the high sensitivity of the surface-enhanced Raman spectroscopy, have brought new insights into our knowledge of abnormal protein aggregation. In this review, we order and summarize all nano- and micro-spectroscopic marker bands related to abnormal aggregation. Each part presents the physical principles of each particular spectroscopic technique listed above and a concise description of all spectral markers detected with these techniques in the spectra of neurodegenerative proteins and their model systems. Finally, a section concerning the application of multivariate data analysis for extraction of the spectral marker bands is included

Stearoyl-CoA desaturase 1 activity determines the maintenance of DNMT1-mediated DNA methylation patterns in pancreatic β\beta-Cells

Metabolic stress, such as lipotoxicity, affects the DNA methylation profile in pancreatic β-cells and thus contributes to β-cell failure and the progression of type 2 diabetes (T2D). Stearoyl-CoA desaturase 1 (SCD1) is a rate-limiting enzyme that is involved in monounsaturated fatty acid synthesis, which protects pancreatic β-cells against lipotoxicity. The present study found that SCD1 is also required for the establishment and maintenance of DNA methylation patterns in β-cells. We showed that SCD1 inhibition/deficiency caused DNA hypomethylation and changed the methyl group distribution within chromosomes in β-cells. Lower levels of DNA methylation in SCD1-deficient β-cells were followed by lower levels of DNA methyltransferase 1 (DNMT1). We also found that the downregulation of SCD1 in pancreatic β-cells led to the activation of adenosine monophosphate-activated protein kinase (AMPK) and an increase in the activity of the NAD-dependent deacetylase sirtuin-1 (SIRT1). Furthermore, the physical association between DNMT1 and SIRT1 stimulated the deacetylation of DNMT1 under conditions of SCD1 inhibition/downregulation, suggesting a mechanism by which SCD1 exerts control over DNMT1. We also found that SCD1-deficient β-cells that were treated with compound c, an inhibitor of AMPK, were characterized by higher levels of both global DNA methylation and DNMT1 protein expression compared with untreated cells. Therefore, we found that activation of the AMPK/SIRT1 signaling pathway mediates the effect of SCD1 inhibition/deficiency on DNA methylation status in pancreatic β-cells. Altogether, these findings suggest that SCD1 is a gatekeeper that protects β-cells against the lipid-derived loss of DNA methylation and provide mechanistic insights into the mechanism by which SCD1 regulates DNA methylation patterns in β-cells and T2D-relevant tissues

Kardiomiopatia przerostowa z koniuszkowym tętniakiem lewej komory

We present a case of 68 year-old woman with previously diagnosed hypertrophic cardiomyopathy complaining of progressive dyspnoe after the acute coronary syndrome. The echocardiogram confirmed hypertrophic hypertrophy and revealed an aneurysm in the apical portion of the left ventricle (LV). The Doppler-echocardiography showed two intra-LV gradients: midventricular and subaortic, with systolic anterior motion of anterior mitral leaflet. Kardiol Pol 2011; 69, 9: 963–96

Wartość prognostyczna scyntygrafii perfuzyjnej mięśnia sercowego metodą SPECT z użyciem 99mTc-MIBI w grupie pacjentów po przebytym w przeszłości zawale serca

Introduction. Data describing prognostic utility of SPECT (single-photon emission computed tomography) among patients with history of myocardial infarction (MI) is scarce. We aimed to compare prognostic value of SPECT scans in patients with vs. without history of myocardial infarction (MI) prior to perfusion imaging. Material and methods. 151 consecutive patients with history of myocardial infarction, who underwent exercise 99mTc-MIBI SPECT examination were enrolled in the study (group 1). Next, based on clinical and demographic baseline characteristics, SPECT result and duration of follow-up period, a matching control subgroup (group 2) of 151 patients without history of MI was formed. During a mean follow-up of 60 ± 15 months in group 1 and a mean follow-up of 60 ± 13.5 months in group 2, we analyzed prevalence of cardiovascular events rates (cardiac deaths, myocardial infarctions and cardiac revascularizations). Results. We observed the following rates of cardiac deaths, myocardial infarctions and cardiac revascularizations: 4%, 15% and 26%, respectively in group 1 and 1%, 13% and 17%, respectively in group 2. In patients without history of MI (group 2) presence of mixed perfusion defects on SPECT was associated with significantly higher rate of myocardial infarctions (p = 0.05), hospitalizatons (p = 0.0001) and revascularizations (p = 0.0002). Fixed perfusion defects were associated with significantly higher occurrence of deaths (p = 0.012) and hospitalizations (p = 0.02), but not myocardial infarctions (p = 0.52) or cardiac revascularizations (p = 0.10). In contrast, among patients with history of MI (group 1) there was no statistically significant relationship between any type of perfusion defects and cardiovascular events. Conclusions. Long term follow-up demonstrates that prognostic value of 99mTc-MIBI SPECT is different in patients with and without history of MI.Wstęp. W dostępnej literaturze brakuje badań służących ocenie przydatności prognostycznej scyntygrafii perfuzyjnej mięśnia sercowego SPECT (tomografia emisyjna pojedynczego fotonu) w grupie chorych z wywiadem przebytego zawału serca. Celem opisanego badania jest porównanie wartości prognostycznej SPECT w grupie pacjentów z wywiadem zawału serca przed badaniem SPECT oraz bez takiego wywiadu. Materiał i metody. Do badania włączono 151 kolejnych pacjentów z wywiadem zawału serca, u których wykonano badanie SPECT z użyciem 99mTc-MIBI. Następnie, na podstawie charakterystyki klinicznej i demograficznej oraz długości okresu obserwacji dobrano, grupę kontrolną 151 pacjentów poddanych badaniu SPECT, bez wywiadu wcześniejszego zawału serca. Wyniki. Po średnim okresie obserwacji 60 ± 15 miesięcy (grupa 1) i 60 ± 13,5 miesiąca (grupa 2) częstość wystąpienia punktów końcowych, takich jak zgon z przyczyn sercowo-naczyniowych, zawał serca, rewaskularyzacja, wyniosła odpowiednio 4%, 15% i 26% w grupie 1 oraz 1%, 13%, 17% w grupie 2. Wśród pacjentów bez wywiadu zawału serca (grupa 2) obecność mieszanych zaburzeń perfuzji w badaniu SPECT wiązała się z istotnie większą częstością zawałów serca (p =0,05), hospitalizacji (p = 0,0001) oraz rewaskularyzacji (p = 0,0002). Utrwalone zaburzenia perfuzji wiązały się z istotnie większą liczbą zgonów z przyczyn sercowo-naczyniowych, liczbą hospitalizacji (p=0,02), ale nie zawałów serca (p = 0,52) ani rewaskularyzacji (p = 0,10). Natomiast wśród pacjentów bez wywiadu zawału serca (grupa 1) nie zaobserwowano istotnej statystycznie zależności między typem zaburzeń perfuzji a występowaniem powyższych punktów końcowych. Wnioski. W długoterminowej obserwacji wykazano różnicę w zakresie wartości rokowniczej badania SPECT między pacjentami po przebytym zawale serca oraz bez wywiadu zawału

Combined analytical approach empowers precise spectroscopic interpretation of subcellular components of pancreatic cancer cells

The lack of specific and sensitive early diagnostic options for pancreatic cancer (PC) results in patients being largely diagnosed with late-stage disease, thus inoperable and burdened with high mortality. Molecular spectroscopic methodologies, such as Raman or infrared spectroscopies, show promise in becoming a leader in screening for early-stage cancer diseases, including PC. However, should such technology be introduced, the identification of differentiating spectral features between various cancer types is required. This would not be possible without the precise extraction of spectra without the contamination by necrosis, inflammation, desmoplasia, or extracellular fluids such as mucous that surround tumor cells. Moreover, an efficient methodology for their interpretation has not been well defined. In this study, we compared different methods of spectral analysis to find the best for investigating the biomolecular composition of PC cells cytoplasm and nuclei separately. Sixteen PC tissue samples of main PC subtypes (ductal adenocarcinoma, intraductal papillary mucinous carcinoma, and ampulla of Vater carcinoma) were collected with Raman hyperspectral mapping, resulting in 191,355 Raman spectra and analyzed with comparative methodologies, specifically, hierarchical cluster analysis, non-negative matrix factorization, T-distributed stochastic neighbor embedding, principal components analysis (PCA), and convolutional neural networks (CNN). As a result, we propose an innovative approach to spectra classification by CNN, combined with PCA for molecular characterization. The CNN-based spectra classification achieved over 98% successful validation rate. Subsequent analyses of spectral features revealed differences among PC subtypes and between the cytoplasm and nuclei of their cells. Our study establishes an optimal methodology for cancer tissue spectral data classification and interpretation that allows precise and cognitive studies of cancer cells and their subcellular components, without mixing the results with cancer-surrounding tissue. As a proof of concept, we describe findings that add to the spectroscopic understanding of PC