Organic amendments improve salinity-induced osmotic and oxidative stress tolerance in Okra (Abelmoschus esculentus (L.)Moench).

AIMS: Salinity adversely affects okra [Abelmoschus esculentus (L.) Moench] plants by inducing osmotic and oxidative stresses. This study was designed to enhance salinity-induced osmotic and oxidative stress tolerance in okra plants by applying organic amendments. METHODS: The effects of different organic amendments (municipal solid waste compost, farmyard manure (FYM) and press mud) on osmotic potential, water use efficiency, activities of antioxidant enzymes, total soluble sugar, total soluble proline, total soluble protein and malondialdehyde (MDA) contents of okra plants grown under saline conditions (50 mM sodium chloride) were evaluated in a pot experiment. The organic amendments were applied each at the rate of 5% and 10% per pot or in various combinations (compost + FYM, FYM + press mud and compost + press mud each at the rate of 2.5% and 5% per pot). RESULTS: As compared to control, high total soluble sugar (60.41), total soluble proline (33.88%) and MDA (51%) contents and increased activities of antioxidant enzymes [superoxide dismutase (83.54%), catalase (78.61%), peroxidase (53.57%] in salinity-stressed okra plants, were indicative of oxidative stress. Salinity significantly reduced the osmotic potential (41.78%) and water use efficiency (4.75%) of okra plants compared to control. Under saline conditions, 5% (farmyard manure + press mud) was the most effective treatment, which significantly improved osmotic potential (27.05%), total soluble sugar (4.20%), total soluble protein (73.62%) and total soluble proline (23.20%) contents and superoxide dismutase activity (32.41%), compared to saline soil. Application of 2.5% (FYM + press mud), 5% press mud, and 10% compost significantly reduced MDA content (27%) and improved activities of catalase (38.64%) and peroxidase (48.29%), respectively, compared to saline soil, thus facilitated to alleviate oxidative stress in okra plants. CONCLUSIONS: Using organic amendments (municipal solid waste compost, farmyard manure and press mud) was a cost-effective approach to improve salinity-induced osmotic and oxidative stress tolerance in okra plants.

Vibrational Corrections to NMR Spin-Spin Coupling Constants from Relativistic Four-Component DFT Calculations.

Zero-point vibrational (ZPV) corrections to the nuclear spin-spin coupling constants have been calculated using four-component Dirac-Kohn-Sham DFT for H2X (where X = O, S, Se, Te, Po), XH3 (where X = N, P, As, Sb, Bi), and XH4 (where X = C, Si, Ge, Sn, and Pb) molecules and for HC≡CPbH3. The main goal was to study the influence of relativistic effects on the ZPV corrections and thus results calculated at relativistic and nonrelativistic approaches have been compared. The effects of relativity become notable for the ZPV corrections to the spin-spin coupling constants for compounds with lighter elements (selenium and germanium) than for the spin-spin coupling constants themselves. In the case of molecules containing heavier atoms, for instance BiH3 and PbH4, relativistic effects play a crucial role on the results and approximating ZPV corrections by the nonrelativistic results may lead to larger errors than omitting ZPV corrections altogether.

The Role of Oxidative Stress in the Risk of Cardiovascular Disease and Identification of Risk Factors Using AIP and Castelli Atherogenicity Indicators in Patients with PCOS.

Polycystic ovarian syndrome (PCOS) is one of the most common endocrinopathies in females of reproductive age and may affect 5-14% of women. In women with PCO syndrome, metabolic disorders such as insulin resistance, hyperinsulinemia, obesity, diabetes mellitus, and other elements of metabolic syndrome may occur. Patients with PCOS often have overweight and obesity, especially abdominal obesity, which is one of the risk factors for developing atherosclerosis. The atherogenicity indicators of AIP (atherogenic index of plasma) and Castelli's index are used to assess the risk of developing atherosclerosis. Studies have shown an increase in the concentration and activity of oxidative stress markers in patients with PCOS compared to women without the disease. The aim of the present study was to evaluate oxidative stress parameters in patients with PCOS in relation to insulin resistance, BMI, and hyperandrogenemia and to correlate them with cardiovascular risk parameters. Conclusions: The severity of oxidative stress in women with PCOS correlates with exposure to cardiovascular diseases. The assessment of additional cardiovascular disease (CVD) parameters is useful in identifying the risk groups for cardiometabolic disease among PCOS patients. When additional risk factors such as hyperandrogenism and insulin resistance (IR) are present in patients with PCOS, it is reasonable to include preventive examinations early. It is also important to evaluate lipidograms, which will make it possible to determine indicators of atherogenicity. Patients with PCOS and IR are at particular risk for cardiovascular complications. PCOS should be considered an important risk factor for CVD, which occurs independently of the occurrence of obesity. This factor is related to the important role of insulin resistance, which occurs independently of obesity. Atherogenic factors (AIP and Castelli index) are useful additional parameters to assess the risk of cardiometabolic disease in PCOS patients, especially among groups with insulin resistance. The early detection of risk factors should be an integral part of the care of PCOS patients. In laboratory studies of women with PCOS, TG, TChol, HDL-c and LDL-c levels, and glutathione peroxidase (GPx) activity were most clearly correlated with exposure to cardiovascular disease.

Neutrophil infiltration combined with necrosis in the primary tumor is a useful prognostic indicator for three-year disease-free survival time in patients with colorectal cancer.

Histopathological evaluation plays a key role in the diagnosis of colorectal cancer (CRC). Tumor-related local inflammation is regarded as a novel prognostic parameter. Neutrophils constitute one of the main types of inflammatory cells. The aim of the present study was to evaluate the prognostic value of intratumoral tumor-associated neutrophils (intraTANs), stromal TANs (stromaTANs) and necrosis, as well as their combined parametric value in formalin-fixed paraffin-embedded tissue sections from patients with CRC. For this purpose, a retrospective study of 160 patients with CRC who underwent surgery was conducted. The association of intraTANs, stromaTANs, necrosis and their combined parametric value with the clinicopathological features of patients with CRC was examined. The Kaplan-Meier method and the log-rank test were used to compare survival curves. To identify independent prognostic factors, uni- and multivariate Cox proportional hazards regression models were used. StromaTANs were associated with lymph node metastasis (P=0.049) and tumor deposits (P=0.041). In addition, necrosis was found to be associated with venous (P=0.003), lymphatic (P=0.007) and perineural (P=0.015) invasion, as well as with lymph node metastasis (P=0.033), the number of invaded lymph nodes (P=0.012), and lymph node pouch invasion (P=0.043). Furthermore, necrosis was found to be associated with the white blood cell count (P=0.030), neutrophil count (P=0.011), the combined neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (NLR-PLR) (P=0.038), and the combined platelet and NLR (PLT-NLR status) (P=0.030), as well as with the serum carcinoembryonic antigen (CEA) levels following surgery (P=0.011) and the monocyte-to-lymphocyte ratio (P=0.023). The combined parametric value was found to be associated with pT stage (P=0.049), venous (P=0.034) and lymphatic (P=0.026) invasion, and with serum CEA levels prior to surgery (P=0.029). The analysis of the 3-year disease-free survival (DFS) time revealed that tumor growth [hazard ratio (HR), 2.070; 95% CI, 1.837-3.808; P=0.003] and the combined parametric value (intraTANs, stromaTANs and necrosis, HR, 1.577; 95% CI, 1.372-3.032; P=0.028) were independent factors for patients with CRC. Taken together, the findings of the present study demonstrated that the combined value of neutrophils and necrosis examined in the cancerous tissue may be used as a prognostic factor for the 3-year DFS time in patients with CRC.

Assessment of the Parameters of Oxidative Stress Depending on the Metabolic and Anthropometric Status Indicators in Women with PCOS.

Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies in females of reproductive age. In women with PCOS, metabolic disorders such as insulin resistance (IR), hyperinsulinemia, obesity, diabetes mellitus, and other elements of metabolic syndrome are likely to occur. Studies have shown an increase in the concentration and activity of oxidative stress (OS) markers in patients with PCOS, compared to that in unaffected women. The aim of this study was to evaluate the parameters of OS in PCOS and their activity in relation to women without menstrual disorders with a normal body weight. Then, we compared malonodialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), including overweight and obesity, hyperandrogenemia, and IR in the PCOS group. The study included 35 women aged 18-46, hospitalized for menstrual disorders in the form of infrequent menstruation. In 26 women, PCOS was diagnosed on the basis of the Rotterdam Criteria; these patients qualified for the study group. The control group (n = 21) consisted of patients without menstrual disorders and without PCOS in an ultrasound examination. Patients were diagnosed between the 2nd and 5th day of the cycle. The parameters of OS were analyzed and compared with the anthropometric parameters and the lipid profile of the patients. Enzymatic activity of GPx, CAT, SOD, and MDA levels was determined in both groups. MDA levels and CAT activity differed significantly between the groups. There was a decrease in MDA levels in the IR group and the involvement of GPx in the excess weight and obesity and IR group accompanied by an increase in hip circumference. It therefore seems that IR may be the main risk factor to exposure to OS in patients with PCOS, independent from obesity. In addition, GPx is involved in every step in the development of the pathological condition in PCOS.

Relativistic Four-Component DFT Calculations of Vibrational Frequencies.

We investigate the effect of relativity on harmonic vibrational frequencies. Density functional theory (DFT) calculations using the four-component Dirac-Coulomb Hamiltonian have been performed for 15 hydrides (H2X, X = O, S, Se, Te, Po; XH3, X = N, P, As, Sb, Bi; and XH4, X = C, Si, Ge, Sn, Pb) as well as for HC≡CPbH3. The vibrational frequencies have been calculated using finite differences of the molecular energy with respect to geometrical distortions of the nuclei. The influences of the choice of basis set, exchange-correlation functional, and step length for the numerical differentiation on the calculated harmonic vibrational frequencies have been tested, and the method has been found to be numerically robust. Relativistic effects are noticeable for the heavier congeners H2Te and H2Po, SbH3 and BiH3, and SnH4 and PbH4 and are much more pronounced for the vibrational modes with higher frequencies. Spin-orbit effects constitute a very small fraction of the total relativistic effects, except for H2Te and H2Po. For HC≡CPbH3 we find that only the frequencies of the modes with large contributions from Pb displacements are significantly affected by relativity.

The role of the electrokinetic charge of neurotrophis-based nanocarriers: protein distribution, toxicity, and oxidative stress in in vitro setting.

BACKGROUND: The rational chemical design of nanoparticles can be readily controlled and optimized by quantitatively studying protein adsorption at variously charged polymer carriers, determining their fate in biological fluids. We manufactured brain-derived neurotrophic factor (BDNF) -based electrostatic nanocomplexes with a different type of dendrimer core (anionic or cationic), encapsulated or not in polyethylene glycol (PEG), and studied their physicochemical properties and behavior in a biological setting. We investigated whether the electrokinetic charge of dendrimer core influences BDNF loading and desorption from the nanoparticle and serves as a determinant of nanoparticles' behavior in in vitro setting, influencing mitochondrial dysfunction, lipid peroxidation, and general nanoparticles' cellular toxicity. RESULTS: We found that the electrokinetic charge of the dendrimer core influences nanoparticles in terms of BDNF release profile from their surfaces and their effect on cell viability, mitochondrial membrane potential, cell phenotype, and induction of oxidative stress. The electrostatic interaction of positively charged core of nanoparticles with cell membranes increases their cytotoxicity, as well as serious phenotype alterations compared to negatively charged nanoparticles core in neuron-like differentiated human neuroblastoma cells. Moreover, PEG adsorption at nanoparticles with negatively charged core presents a distinct decrease in metabolic cell activity. On the contrary, charge neutralization due to PEG adsorption on the surface of nanoparticles with positively charged core does not reduce their cytotoxicity, makes them less biocompatible with differentiated cells, and presumably shows non-specific toxicity. CONCLUSIONS: The surface charge transformation after adsorption of protein or polyelectrolyte during nanocarriers formulation has an important role not only in designing nanomaterials with potent neuroprotective and neuroregenerative properties but also in applying them in a cellular environment.

Tumor-infiltrating lymphocytes in tissue material combined with systemic lymphocyte inflammation in patients with colorectal cancer.

The efficacy of cancer immunotherapy has been actively explored in the treatment of various malignant neoplasms of the gastrointestinal tract. In light of recent reports, the present study aimed to investigate the combination of the absolute lymphocyte count (ALC), percentage of tumor-infiltrating lymphocyte (TILs) and tumor progression status in patients with colorectal cancer (CRC) who underwent surgery. The medical records of 160 patients diagnosed with CRC were retrospectively reviewed. TILs were determined as a percentage of mononuclear inflammatory cells in the total intratumoral or stromal area as determined in five high power fields (magnification, x200-400), at the invasive front and in the centre of the tumour. Blood samples were obtained within 3 days prior to and 7 days following the surgical treatment. The assessment of the TIL percentage was performed in the tissue at the invasive front and in the centre of the primary tumour mass in combination with the determination of ALC in whole blood samples. The samples were obtained prior to and after surgery from patients with CRC, and the tumour progression status was also assessed (TILs/ALC/tumour progression status). A significant association was observed between the percentage of TILs in the main mass of tumour and the tumour size (P=0.031), the pT stage (P=0.049) and the incidence of necrosis (P=0.037) following surgery. The histological type was associated with the evaluated combined parameters prior to surgery (P=0.046). Lymph node pouch invasion was associated with TILs at the invasive front of tumour and with ALC prior to and after surgery (P=0.006 and P=0.037). Furthermore, the data indicated that the percentage of TILs located on the invasive front and centre of the tumour, and the ALC prior to and following surgery correlated with the treatment status (P=0.032, P=0.018, P≤0.001 and P≤0.001). A significant association was noted between eight features and evaluated combined parameters following surgery. These included the tumour size (P=0.021), TNM stage (P<0.001), tumour deposits (P=0.001), incidence of necrosis (P=0.042) and lymph node metastasis (P<0.001). Furthermore, the degree of invasion of venous (P<0.001), lymphatic (P<0.001) and perineural (P<0.001) sites was also significantly associated with TILs, ALC obtained after surgical treatment and tumor progression status. The data demonstrated that local and systemic chronic inflammation was associated with tumour progression in patients with CRC.

The intensification of anticancer activity of LFM-A13 by erythropoietin as a possible option for inhibition of breast cancer.

Recombinant human erythropoietin (Epo) is an effective and convenient treatment for cancer-related anaemia. In our study for the first time, we evaluated the effect of simultaneous use of Epo and Bruton's tyrosine kinase (BTK) inhibitor LFM-A13 on the viability and tumour development of breast cancer cells. The results demonstrated that Epo significantly intensifies the anticancer activity of LFM-A13 in MCF-7 and MDA-MB-231. The featured therapeutic scheme efficiently blocked the tumour development in zebrafish experimental cancer model. Epo and LFM-A13 administered together resulted in effective cell killing, accompanied by attenuation of the BTK signalling pathways, loss of mitochondrial membrane potential (MMP), accumulation of apoptotic breast cancer cells with externalised PS, a slight increase in phase G0/G1 and a reduction in cyclin D1 expression. Simultaneous use of Epo with LFM-A13 inhibited early stages of tumour progression. This therapeutic scheme may be rationale for further possible research.

Monocyte-to-lymphocyte ratio as a prognostic factor in peripheral whole blood samples of colorectal cancer patients.

BACKGROUND: Colorectal cancer is the third most common malignancy worldwide. Therefore, it is critically important to identify new useful markers that can be easily obtained in routine practice. Inflammation is a crucial issue in the pathogenesis and development of cancer. AIM: To evaluate the prognostic value of absolute monocyte count, monocyte to lymphocyte ratio (MLR), the combination of neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio (NLR-PLR), and combined platelet and neutrophil-to-lymphocyte ratio (PLT-NLR) in peripheral blood samples of patients with colorectal cancer undergoing surgery. METHODS: We conducted a retrospective study of 160 patients with colorectal cancer who underwent surgery, and 42 healthy controls. The status of absolute monocyte count, MLR, NLR-PLR and PLT-NLR was calculated on the basis of blood samples obtained before and after surgery. Haematologic factors were examined in correlation with the type of tumour growth, tumour size, histological type, percentage of mucinous component, grade of malignancy, Tumour-Node-Metastasis stage, venous, lymphatic and perineural invasion of cancer cells, status of lymph node invasion and the presence of cancer cell deposits. The Kaplan-Meier method and the long-rank test were used to compare survival curves. To determine independent prognostic factors, univariate and multivariate Cox proportional hazards regression models were applied. RESULTS: The PLT-NLR status was correlated with tumour size and the presence of perineural invasion (P = 0.015; P = -0.174, P = 0.037). Moreover, high NLR-PLR and PLR-NLR ratios in the blood samples obtained after surgery were positively associated with histological type of cancer and percentage of the mucinous component (NLR-PLR: P = 0.002; P = 0.009; PLR-NLR status: P = 0.002; P = 0.007). The analysis of 5-year disease-free survival showed that the MLR of whole blood obtained after surgery [HR = 2.903, 95%CI: (1.368-6.158), P = 0.005] and the status of lymph node metastasis [HR = 0.813, 95%CI: (0.653-1.013), P = 0.050] were independent prognostic factors in colorectal cancer patients. CONCLUSION: The postoperative MLR in whole blood samples can be used as an independent prognostic factor in patients diagnosed with colorectal cancer.

Pre- and postoperative neutrophil and lymphocyte count and neutrophil-to-lymphocyte ratio in patients with colorectal cancer.

Colorectal cancer (CRC) is one of the most common malignant cancers worldwide. Patients with CRC are diagnosed based on various predictors, including performance status, clinicopathological factors and TNM classification. The aim of the present study was to analyze the neutrophil and lymphocyte counts, as well as the neutrophil-to-lymphocyte ratio (NLR) in pre- and postoperative blood samples of patients with CRC in correlation with specific anatomical variables and disease- free survival (DFS). The variables pre- and postoperative neutrophil count (preNEU and postNEU, respectively), lymphocyte count and NLR were significantly higher in cancer patients than those noted in healthy subjects (all P<0.001). PreNEU count correlated with tumor size, necrosis and tumor budding (R=0.204, P=0.014; R=0.189, P=0.023; R=-0.174, P=0.036, respectively). Moreover, postNEU was associated only with the histological type (R=0.174; P=0.047). The PreLYMPH count was correlated with distant metastasis (R=-0.153, P=0.046). PreNLR and postNLR were associated with the expression of various histological markers of disease progression. Analysis of DFS indicated that the postNEU count in the low group exhibited a tendency to lower DFS duration, although the results were not significant (P=0.055). In conclusion, the present study indicated a significant correlation between the factors analyzed in blood samples of CRC patients and the disease progression markers.

Insulin receptor substrate 1 may play divergent roles in human colorectal cancer development and progression.

BACKGROUND: Despite effective prevention and screening methods, the incidence and mortality rates associated with colorectal cancer (CRC) are still high. Insulin receptor substrate 1 (IRS-1), a signaling molecule involved in cell proliferation, survival and metabolic responses has been implicated in carcinogenic processes in various cellular and animal models. However, the role of IRS-1 in CRC biology and its value as a clinical CRC biomarker has not been well defined. AIM: To evaluate if and how IRS-1 expression and its associations with the apoptotic and proliferation tumor markers, Bax, Bcl-xL and Ki-67 are related to clinicopathological features in human CRC. METHODS: The expression of IRS-1, Bax, Bcl-xL and Ki-67 proteins was assessed in tissue samples obtained from 127 patients with primary CRC using immunohistochemical methods. The assays were performed using specific antibodies against IRS-1, Bax, Bcl-xL, Ki-67. The associations between the expression of IRS-1, Bax, Bcl-xL, Ki-67 were analyzed in relation to clinicopathological parameters, i.e., patient age, sex, primary localization of tumor, histopathological type, grading, staging and lymph node spread. Correlations between variables were examined by Spearman rank correlation test and Fisher exact test with a level of significance at P < 0.05. RESULTS: Immunohistochemical analysis of 127 CRC tissue samples revealed weak cytoplasmatic staining for IRS-1 in 66 CRC sections and strong cytoplasmatic staining in 61 cases. IRS-1 expression at any level in primary CRC was associated with tumor grade (69% in moderately differentiated tumors, G2 vs 31% in poorly differentiated tumors, G3) and with histological type (81.9% in adenocarcinoma vs 18.1% in adenocarcinoma with mucosal component cases). Strong IRS-1 positivity was observed more frequently in adenocarcinoma cases (95.1%) and in moderately differentiated tumors (85.2%). We also found statistically significant correlations between expression of IRS-1 and both Bax and Bcl-xL in all CRC cases examined. The relationships between studied proteins were related to clinicopathological parameters of CRC. No significant correlation between the expression of IRS-1 and proliferation marker Ki-67, excluding early stage tumors, where the correlation was positive and on a high level (P = 0.043, r = 0.723). CONCLUSION: This study suggests that IRS-1 is co-expressed with both pro- and antiapoptotic markers and all these proteins are more prevalent in more differentiated CRC than in poorly differentiated CRC.

NMR Spin-Spin Coupling Constants Derived from Relativistic Four-Component DFT Theory-Analysis and Visualization.

An unambiguous assignment of coupling pathways plays an important role in the description and rationalization of NMR indirect spin-spin coupling constants (SSCCs). Unfortunately, the SSCC analysis and visualization tools currently available to quantum chemists are restricted to nonrelativistic theory. Here, we present the theoretical foundation for novel relativistic SSCC visualization techniques based on analysis of the SSCC densities and the first-order current densities induced by the nuclear magnetic dipole moments. Details of the implementation of these techniques in the ReSpect program package are discussed. Numerical assessments are performed on through-space SSCCs, and we choose as our examples the heavy-atom Se-Se, Se-Te, and Te-Te coupling constants in three similar molecules for which experimental data are available. SSCCs were calculated at the nonrelativistic, scalar relativistic, and four-component relativistic density functional levels of theory. Furthermore, with the aid of different visualization methods, we discuss the interpretation of the relativistic effects, which are sizable for Se-Se, very significant for Se-Te, and cannot be neglected for Te-Te couplings. A substantial improvement of the theoretical SSCC values is obtained by also considering the molecular properties of a second conformation.

Tumor-infiltrating lymphocytes in primary tumors of colorectal cancer and their metastases.

The presence of tumor cells in the large intestine stimulates hypoxia and local inflammatory mediators that activate numerous inflammatory cells, including a diverse lymphoid tumor cell population. The aim of the present study was to evaluate tumor-infiltrating lymphocytes (TILs) located in the invasive primary tumor, surrounding deposits of tumor cells and those present in distal metastatic cells in the liver of patients with colorectal cancer. Furthermore, the correlation of TILs with anatomical parameters was assessed. The study group included 123 patients with primary tumor colorectal cancer without distant metastasis, 25 cases with deposits of colorectal cancer cells and 15 cases of colorectal cancer liver metastasis. TILs were assessed in tissues stained with hematoxylin-eosin using light microscopy and evaluated by two independent pathologists blinded to the clinical information. Infiltration of TILs in the invasive front of primary tumor was stronger compared with those surrounding deposits of cancer cells and liver metastases (P<0.001). TILs in the invasive front of primary tumor masses were associated with various variables linked with tumor progression and inflammatory cell infiltrate. TILs distributed around the deposits of cancer cells were associated with postoperative treatment; however, those localized in the invasive front of liver metastases were correlated with preoperative therapy. In conclusion, TILs assessment in primary tumors of colorectal cancer, surrounding deposits of tumor cells and in the metastatic cells in the liver may be helpful in understanding the role of these cells in the organization of immune response.

Enhanced relativistic-electron beam collimation using two consecutive laser pulses.

The double laser pulse approach to relativistic electron beam (REB) collimation in solid targets has been investigated at the LULI-ELFIE facility. In this scheme two collinear laser pulses are focused onto a solid target with a given intensity ratio and time delay to generate REBs. The magnetic field generated by the first laser-driven REB is used to guide the REB generated by a second delayed laser pulse. We show how electron beam collimation can be controlled by properly adjusting the ratio of focus size and the delay time between the two pulses. We found that the maximum of electron beam collimation is clearly dependent on the laser focal spot size ratio and related to the magnetic field dynamics. Cu-Kα and CTR imaging diagnostics were implemented to evaluate the collimation effects on the respectively low energy (≤100 keV) and high energy (≥MeV) components of the REB.

E-selectin gene haplotypes are associated with the risk of myocardial infarction.

INTRODUCTION: Endothelial dysfunction is one of the most important factors implicated in the pathogenesis of coronary artery disease (CAD). The aim of this study was to investigate the association of the E-selectin gene (SELE) with CAD and CAD-related traits using tagging polymorphisms. MATERIAL AND METHODS: A total of 379 Polish patients who had undergone angiography were included: 261 patients with angiographically documented CAD, 202 CAD patients without myocardial infarction (CAD/MI(-) group) and 59 patients with myocardial infarction (CAD/MI(+) group) as well as 118 healthy control subjects (non-CAD). Eight tagging single nucleotide polymorphisms (SNPs) in the SELE gene were selected using genotype data from HapMap. Genotyping was performed using PCR-RFLP and PCR-DHPLC methods. RESULTS: The most common SELE haplotype in this analysis ([C;G;T;C;G;T], 31.2%) showed a negative association with myocardial infarction (MI) (CAD/MI(+) vs. non-CAD) under the additive (p = 0.001), dominant (p = 0.006) and recessive (p = 0.012) model. Two other haplotypes ([C;G;C;C;A;C], [C;A;C;A;G;T], 5.73% and 18.1%, respectively) were also negatively associated with MI under the additive and dominant model. We also found two haplotypes ([T;G;T;C;G;T], [C;G;C;C;A;T], 1.52% and 6.71%, respectively) associated with the risk for MI (CAD/MI(+) vs. CAD/MI(-)), acting in both additive (p = 0.04, p = 0.007, respectively) and dominant (p = 0.04, p = 0.004, respectively) manner. There was no association with either CAD/MI(-) or with severity of CAD expressed as the number of vessels involved. CONCLUSIONS: Our results suggest that SELE is one of the independent genetic factors modifying the risk of myocardial infarction.

Prognostic significance of inflammatory cell response in patients with colorectal cancer.

Cancer cells are characterized by a low antigenic immunogenicity, a rapid growth and an immunosuppressive effect on the extracellular matrix. These properties induce a weak immune response in colorectal cancer (CRC) carcinogenesis. It is therefore crucial to determine the composition of the inflammatory mass, including neutrophils, macrophages and eosinophils in the tumor tissue of patients with CRC, and to analyze other clinicopathological parameters. The present study included 144 patients diagnosed with CRC. Tissue samples obtained from routine histopathological diagnosis were stained with hematoxylin and eosin. Inflammatory cells were assessed in the invasive front and in the center of the tumor by light microscopy under a high-power magnification. The percentage of neutrophils in the invasive front was significantly higher compared with that in the center of the tumor mass (P<0.01). Macrophages and eosinophils were present in the invasive front and in the center of tumor mass in most cases. The presence of neutrophils, macrophages and eosinophils was correlated with various clinicopathological features. Patients with macrophages present in the center of tumor mass had longer disease-free survival time (P=0.041). In conclusion, the present study demonstrated that the inflammatory cell infiltrate served a significant role in the immune response of patients with CRC. It should be noted that the presence of macrophages localized in the stroma of the central part of the primary tumor mass was associated with the survival time of patients with CRC.

Analysis of Base Monomer Elution from 3 Flowable Bulk-Fill Composite Resins Using High Performance Liquid Chromatography (HPLC).

BACKGROUND The aim of this study was to evaluate the elution of BisGMA, UDMA, TEGDMA, and HEMA monomers from flowable bulk fill composite resins with different resin matrix compositions, polymerized in 4-mm-thick layers, into 3 elution media. MATERIAL AND METHODS Three bulk-fill (SDR® (SDR), X-tra base (XB) and BEAUTIFIL-Bulk Flowable (BF)) resin-composites were tested. Cylindrical samples were immersed in 100% ethanol, 75% ethanol, and distilled water. The concentrations of the monomers were measured using the HLPC method (Agilent Technologies 1200 Series) after 1 and 24 h, as well as after 3, 7, 14, and 21 days. RESULTS After polymerization of the tested resins, there was elution of the BisGMA, UDMA, TEGDMA, and HEMA monomers from the SDR and BF composites, but none of the tested monomers could be detected eluting from XB. The highest penetrations of the polymerized SDR and BF composites were observed in the 100% ethanol solution. This extraction medium eluted the highest amounts of free monomers. Some eluted monomers were not described in the composites Material Safety Data Sheets. CONCLUSIONS The elution of the residual monomers depended on the resin composition and the materials filler/resin matrix ratio. In composite materials, toxicity assessment should be carried out, and should consider both the material composition as given by the manufacturer, and also the residual monomers that elute from the polymerized material. The elution concentration and time of monomers from composites depended on the solvent used. The highest penetrations of the polymerized SDR and BF composites were observed in the 100% ethanol solution, and this extraction medium eluted the highest amounts of free monomers. The 75% ethanol was a more aggressive medium than water in terms of monomer elution from bulk fill composites.

Squamous cell carcinoma of the breast as a clinical diagnostic challenge.

Squamous cell carcinoma (SqCC) of the breast should be differentiated between the primary skin keratinizing squamous carcinoma and squamous metaplastic cancer. In the current study, the cases of two patients who were diagnosed with SqCC originated from skin and the breast were discussed. A fine-needle aspiration biopsy confirmed the presence of atypical squamous cells. In both cases, the microscopic examination of the surgical specimen revealed a malignant neoplasm differentiated into SqCC characterized by keratinizing cancer cells with abundant eosiphilic cytoplasm with large, hyperchromatic vesicular nuclei. Immunohistochemical studies showed negative for progesterone and estrogen receptors and human epidermal growth factor receptor 2. Moreover, negative expression of cytokeratin 7 and 20 was confirmed. The diagnosis of the both tumors was established based on the detailed analysis of clinical, macroscopical and microscopical information. SqCC localized in the breast is a great diagnostic challenge in pathomorphology and more attention should be paid for analysis of such lesions in daily practice.

Stromal and intraepithelial tumor-infiltrating lymphocytes in colorectal carcinoma.

The local mechanisms of antitumor immune defense determine the development and organization of the tumor microenvironment, and the composition and relative proportions of the inflammatory cell population affect the quality and characteristics of the immune response. The aim of the present study was to conduct a quantitative morphological evaluation of two types of tumor-infiltrating lymphocyte (TILs) populations, including those located in the stroma and intraepithelial cancer structures, in the invasive front and the center of the tumor in patients with colorectal cancer (CRC). The study included 160 patients with CRC who had undergone surgery. The tissue material was stained with hematoxylin and eosin, as used in routine histopathological diagnosis, and the two TIL populations were observed and counted with light microscopy. The relative extent of infiltration of stromal and intraepithelial TILs into the front and center of the primary tumors was similar. The extent of infiltration by stromal TILs was negatively correlated with the morphological features of tumor progression including the cancer infiltration of blood vessels (P=0.016), the invasion of lymph vessels (P=0.007), perineural invasion (P=0.036), lymph node involvement (P=0.047) and distant metastases (P=0.032). The infiltration by intraepithelial TILs was positively correlated with a desmoplastic reaction (P=0.002). Disease-free survival time was statistically shorter in patients without intraepithelial TILs in the center of the primary tumor mass (P=0.049; hazard ratio = 1.45). These results confirm that the infiltration of TILs into the invasive front and center of the tumor in patients with CRC serves an important role in the invasion and progression of the disease, and should be considered in routine histopathological examinations.