Because infection with E. coli commonly cause diarrhea in weaned rabbits, this study was conducte... more Because infection with E. coli commonly cause diarrhea in weaned rabbits, this study was conducted in order to monitor changes in circulating concentrations of haptoglobin (Hp), fibrinogen (Fb) and ceruloplasmin (Cp) during experimental infection weaned rabbits. For this, 18 rabbits from 40 to 45 days, weaned at 4 weeks were used: 12 were inoculated with strain 015: H - E. coli (6.10 7 cfu) and the other 6 were used as negative controls. Blood samples were performed 24 and 72 hours and 7, 11, 18 and 30 days after inoculation in order to determine the concentrations of acute phase proteins (APP) and coliforms were investigated in faecal samples 1, 6, 44, 16, 25 and 31 days later. The excretion of experimental strain began two days after inoculation with 3 rabbits and strengthened in the 3 rd day while mild to severe diarrhea were observed between the 5 th and 21 th days in all rabbits inoculated except 2. In parallel, bacterial excretion gradually decreased. In addition, Hp concentrations were significantly increased at 24 hours after inoculation with E. Coli, reached maximum values on 7 th day (they were multiplied by 9) and remained significantly elevated compared to baseline until 30 th day. By cons, significant variation compared to baseline concentrations and Cp Fb appeared later (respectively the 3 rd and 7 th days), less intense (maximum values obtained on Day 11 was the approximate than doubled) and less durable (normal values were recovered on 30 th and 18 th days, respectively). Increases in all APPs concentrations were associated with intensity of diarrhea. Positive but moderate correlations were obtained between the concentrations of Hp and Cp or Fb whereas Cp concentrations and Fb were strongly correlated. The evolution in time and amplitude changes in levels of these APPs lead to consider the Cp and Fb proteins as positive but slow evolution of the acute phase while 1'Hp can be classified as a fast protein major and allowing early detection of bacterial infections in weaned rabbits before the onset of clinical signs (diarrhea) and fecal coliform excretion.
Current plant-derived anticancer therapeutics aim to reach higher effectiveness, to potentiate ch... more Current plant-derived anticancer therapeutics aim to reach higher effectiveness, to potentiate chemosensitivity and minimize the toxic side effects compared to conventional chemotherapy. Cotinus coggygria Scop. is a herb with high pharmacological potential, widely applied in traditional phytotherapy. Our previous study revealed that leaf aqueous ethanolic extract from C. coggygria exerts in vitro anticancer activity on human breast, ovarian and cervical cancer cell lines. The objective of the present research was to investigate possible molecular mechanisms and targets of the antitumor activity of the extract in breast cancer MCF7 cells through analysis of cell cycle and apoptosis, clonogenic ability assessment, evaluation of the extract genotoxic capacity, characterization of cells thermodynamic properties, and analysis on the expression of genes involved in cellular epigenetic processes. The obtained results indicated that in MCF7 cells C. coggygria extract causes S phase cell cyc...
The approaches of InterCriteria Analysis and Correlation Analysis are applied to a dataset of cal... more The approaches of InterCriteria Analysis and Correlation Analysis are applied to a dataset of calorimetric and statistical parameters obtained from blood plasma proteome thermograms of colorectal cancer patients. The analysis was performed for four individual predefined subsets of calorimetric profiles. Specific interrelations between the studied criteria were identified that were found to differ among the different calorimetric subsets. For three of the subsets the enthalpy of the thermal profiles was in strong consonance with the excess heat capacity of the immunoglobulins assigned thermal transition. For the calorimetric subsets that differed most from the control healthy set a strong interrelation between the excess heat capacities of the main plasma proteins (albumin and immunoglobulins) was additionally evident. Our results demonstrate that these mathematical approaches can complement the analysis of calorimetric datasets generated for a variety of diseases.
International Journal of Biological Macromolecules, 2021
The interactions of graphene oxide (GO), a 2-dimensional nanomaterial with hydrophilic edges, hyd... more The interactions of graphene oxide (GO), a 2-dimensional nanomaterial with hydrophilic edges, hydrophobic basal plane and large flat surfaces, with biological macromolecules, are of key importance for the development of novel nanomaterials for biomedical applications. To gain more insight into the interaction of GO flakes with human serum albumin (HSA), we examined GO binding to HSA in its isolated state and in blood plasma. Calorimetric data reveal that GO strongly stabilizes free isolated HSA against a thermal challenge at low ionic strength, indicating strong binding interactions, confirmed by the drop in ζ-potential of the HSA/GO assemblies compared to bare GO flakes. However, calorimetry also revealed that the HSA-GO molecular interaction is hampered in blood plasma, the ionic strength being particularly important for the interactions. Molecular modelling calculations are in full concert with these experimental findings, indicating a considerably higher binding affinity for HSA to GO in its partially unfolded state, characteristic to low-ionic-strength environment, than for the native protein conformation, observed under physiological conditions. Therefore, for the first time we demonstrate an impeded interaction between HSA and GO nanoflakes in blood plasma, and suggest that the protein is protected from the plausible toxic effects of GO under native conditions.
Breast cancers exhibit different response to drug treatment. In this work, we analyze and compare... more Breast cancers exhibit different response to drug treatment. In this work, we analyze and compare the effect of two anticancer drugs differing in their primary action, miltefosine and cisplatin (cis-Pt), on two different breast cancer (the low—(MCF-7) and high—(MDA-MB-231) metastatic) cell lines, and one normal epithelial (MCF-10A) breast cell lines. The effect of cip-Pt and miltefosine on the thermodynamic behavior of the cancer cell lines was analyzed by differential scanning calorimetry, the cell morphology and viability were determined by optical microscopy and MTT test. We revealed distinct effects of miltefosine and cis-Pt on the thermodynamic behavior and viability of the cancer and normal cells. Importantly, the normal MCF-10A cells were drastically affected by miltefosine, while not by cis-Pt. MDA-MB-231 cell line, on the other hand, is more susceptible to cis-Pt than MCF-7 cells, while both cancer cell lines are equally affected by miltefosine. The drug-associated alteration of the thermal unfolding of the cells constituents correlated with the changes in the cell viability. The altered thermodynamic behavior of the cancer cells upon the drug treatment strongly indicates altered conformations of the proteins in cancer cell membrane and cellular matrix, and the DNA-containing structures.
Differential scanning calorimetry (DSC) gains speed and success in the last decade in the charact... more Differential scanning calorimetry (DSC) gains speed and success in the last decade in the characterization of blood plasma and serum. Numerous publications reveal the potential of this technique to identify calorimetric markers specific for variety of diseases and their staging. In our previous works we have clearly demonstrated that DSC can serve to classify multiple myeloma cases in a number of calorimetric groups whose thermodynamic parameters are strongly affected by the level and isotype of the secreted monoclonal immunoglobulins or free light chains (FLC). In this report we present a case of multiple myeloma with secretion of monoclonal κ FLC (stage III according to ISS classification). High FLC level (about 20% from the total protein content) was found in the patient’s serum that remained persistent for the monitoring period of 1 year. The calorimetric profile of the serum revealed the occurrence of an unusual calorimetric transition at 46-47 °C, unique among nearly 500 multiple myeloma patients studied by us so far. This transition was assigned to unstable monoclonal free light chains that also led to the formation of amorphous aggregates (imaged by atomic force microscopy) in the patient’s serum. Additional studies of patients with similar calorimetric features are needed in order to relate the emergence of the 47 °C transition and protein aggregation to the disease activity status of multiple myeloma or to other pathology.
SummaryPlatelet activation is a complex process in which platelet reorganization takes place asso... more SummaryPlatelet activation is a complex process in which platelet reorganization takes place associated with changes in the cell shape, topology, membrane elasticity and microparticle production. The aim of this study was to investigate the changes/aberrations in the platelet activity, elasticity and morphology in healthy subjects, carriers of A allele of prothrombin G20210A polymorphism. Blood samples from 18 healthy subjects were used for platelet analysis by force-mode atomic force microscopy. Restriction analysis was used to investigate the carriage of G20210A polymorphism in the prothrombin gene. Flow- cytometry was applied to evaluate platelet activation. Young’s modulus of the plasma membranes of platelets derived from healthy subjects, carriers of variant A allele of prothrombin 20210G>A polymorphism (407±69 kPa) is two times higher than the one determined for noncarriers (195.4±48.7 kPa; p<0.05). The background activity of platelets measured as an interrelation of Cd...
Because infection with E. coli commonly cause diarrhea in weaned rabbits, this study was conducte... more Because infection with E. coli commonly cause diarrhea in weaned rabbits, this study was conducted in order to monitor changes in circulating concentrations of haptoglobin (Hp), fibrinogen (Fb) and ceruloplasmin (Cp) during experimental infection weaned rabbits. For this, 18 rabbits from 40 to 45 days, weaned at 4 weeks were used: 12 were inoculated with strain 015: H - E. coli (6.10 7 cfu) and the other 6 were used as negative controls. Blood samples were performed 24 and 72 hours and 7, 11, 18 and 30 days after inoculation in order to determine the concentrations of acute phase proteins (APP) and coliforms were investigated in faecal samples 1, 6, 44, 16, 25 and 31 days later. The excretion of experimental strain began two days after inoculation with 3 rabbits and strengthened in the 3 rd day while mild to severe diarrhea were observed between the 5 th and 21 th days in all rabbits inoculated except 2. In parallel, bacterial excretion gradually decreased. In addition, Hp concentrations were significantly increased at 24 hours after inoculation with E. Coli, reached maximum values on 7 th day (they were multiplied by 9) and remained significantly elevated compared to baseline until 30 th day. By cons, significant variation compared to baseline concentrations and Cp Fb appeared later (respectively the 3 rd and 7 th days), less intense (maximum values obtained on Day 11 was the approximate than doubled) and less durable (normal values were recovered on 30 th and 18 th days, respectively). Increases in all APPs concentrations were associated with intensity of diarrhea. Positive but moderate correlations were obtained between the concentrations of Hp and Cp or Fb whereas Cp concentrations and Fb were strongly correlated. The evolution in time and amplitude changes in levels of these APPs lead to consider the Cp and Fb proteins as positive but slow evolution of the acute phase while 1'Hp can be classified as a fast protein major and allowing early detection of bacterial infections in weaned rabbits before the onset of clinical signs (diarrhea) and fecal coliform excretion.
Current plant-derived anticancer therapeutics aim to reach higher effectiveness, to potentiate ch... more Current plant-derived anticancer therapeutics aim to reach higher effectiveness, to potentiate chemosensitivity and minimize the toxic side effects compared to conventional chemotherapy. Cotinus coggygria Scop. is a herb with high pharmacological potential, widely applied in traditional phytotherapy. Our previous study revealed that leaf aqueous ethanolic extract from C. coggygria exerts in vitro anticancer activity on human breast, ovarian and cervical cancer cell lines. The objective of the present research was to investigate possible molecular mechanisms and targets of the antitumor activity of the extract in breast cancer MCF7 cells through analysis of cell cycle and apoptosis, clonogenic ability assessment, evaluation of the extract genotoxic capacity, characterization of cells thermodynamic properties, and analysis on the expression of genes involved in cellular epigenetic processes. The obtained results indicated that in MCF7 cells C. coggygria extract causes S phase cell cyc...
The approaches of InterCriteria Analysis and Correlation Analysis are applied to a dataset of cal... more The approaches of InterCriteria Analysis and Correlation Analysis are applied to a dataset of calorimetric and statistical parameters obtained from blood plasma proteome thermograms of colorectal cancer patients. The analysis was performed for four individual predefined subsets of calorimetric profiles. Specific interrelations between the studied criteria were identified that were found to differ among the different calorimetric subsets. For three of the subsets the enthalpy of the thermal profiles was in strong consonance with the excess heat capacity of the immunoglobulins assigned thermal transition. For the calorimetric subsets that differed most from the control healthy set a strong interrelation between the excess heat capacities of the main plasma proteins (albumin and immunoglobulins) was additionally evident. Our results demonstrate that these mathematical approaches can complement the analysis of calorimetric datasets generated for a variety of diseases.
International Journal of Biological Macromolecules, 2021
The interactions of graphene oxide (GO), a 2-dimensional nanomaterial with hydrophilic edges, hyd... more The interactions of graphene oxide (GO), a 2-dimensional nanomaterial with hydrophilic edges, hydrophobic basal plane and large flat surfaces, with biological macromolecules, are of key importance for the development of novel nanomaterials for biomedical applications. To gain more insight into the interaction of GO flakes with human serum albumin (HSA), we examined GO binding to HSA in its isolated state and in blood plasma. Calorimetric data reveal that GO strongly stabilizes free isolated HSA against a thermal challenge at low ionic strength, indicating strong binding interactions, confirmed by the drop in ζ-potential of the HSA/GO assemblies compared to bare GO flakes. However, calorimetry also revealed that the HSA-GO molecular interaction is hampered in blood plasma, the ionic strength being particularly important for the interactions. Molecular modelling calculations are in full concert with these experimental findings, indicating a considerably higher binding affinity for HSA to GO in its partially unfolded state, characteristic to low-ionic-strength environment, than for the native protein conformation, observed under physiological conditions. Therefore, for the first time we demonstrate an impeded interaction between HSA and GO nanoflakes in blood plasma, and suggest that the protein is protected from the plausible toxic effects of GO under native conditions.
Breast cancers exhibit different response to drug treatment. In this work, we analyze and compare... more Breast cancers exhibit different response to drug treatment. In this work, we analyze and compare the effect of two anticancer drugs differing in their primary action, miltefosine and cisplatin (cis-Pt), on two different breast cancer (the low—(MCF-7) and high—(MDA-MB-231) metastatic) cell lines, and one normal epithelial (MCF-10A) breast cell lines. The effect of cip-Pt and miltefosine on the thermodynamic behavior of the cancer cell lines was analyzed by differential scanning calorimetry, the cell morphology and viability were determined by optical microscopy and MTT test. We revealed distinct effects of miltefosine and cis-Pt on the thermodynamic behavior and viability of the cancer and normal cells. Importantly, the normal MCF-10A cells were drastically affected by miltefosine, while not by cis-Pt. MDA-MB-231 cell line, on the other hand, is more susceptible to cis-Pt than MCF-7 cells, while both cancer cell lines are equally affected by miltefosine. The drug-associated alteration of the thermal unfolding of the cells constituents correlated with the changes in the cell viability. The altered thermodynamic behavior of the cancer cells upon the drug treatment strongly indicates altered conformations of the proteins in cancer cell membrane and cellular matrix, and the DNA-containing structures.
Differential scanning calorimetry (DSC) gains speed and success in the last decade in the charact... more Differential scanning calorimetry (DSC) gains speed and success in the last decade in the characterization of blood plasma and serum. Numerous publications reveal the potential of this technique to identify calorimetric markers specific for variety of diseases and their staging. In our previous works we have clearly demonstrated that DSC can serve to classify multiple myeloma cases in a number of calorimetric groups whose thermodynamic parameters are strongly affected by the level and isotype of the secreted monoclonal immunoglobulins or free light chains (FLC). In this report we present a case of multiple myeloma with secretion of monoclonal κ FLC (stage III according to ISS classification). High FLC level (about 20% from the total protein content) was found in the patient’s serum that remained persistent for the monitoring period of 1 year. The calorimetric profile of the serum revealed the occurrence of an unusual calorimetric transition at 46-47 °C, unique among nearly 500 multiple myeloma patients studied by us so far. This transition was assigned to unstable monoclonal free light chains that also led to the formation of amorphous aggregates (imaged by atomic force microscopy) in the patient’s serum. Additional studies of patients with similar calorimetric features are needed in order to relate the emergence of the 47 °C transition and protein aggregation to the disease activity status of multiple myeloma or to other pathology.
SummaryPlatelet activation is a complex process in which platelet reorganization takes place asso... more SummaryPlatelet activation is a complex process in which platelet reorganization takes place associated with changes in the cell shape, topology, membrane elasticity and microparticle production. The aim of this study was to investigate the changes/aberrations in the platelet activity, elasticity and morphology in healthy subjects, carriers of A allele of prothrombin G20210A polymorphism. Blood samples from 18 healthy subjects were used for platelet analysis by force-mode atomic force microscopy. Restriction analysis was used to investigate the carriage of G20210A polymorphism in the prothrombin gene. Flow- cytometry was applied to evaluate platelet activation. Young’s modulus of the plasma membranes of platelets derived from healthy subjects, carriers of variant A allele of prothrombin 20210G>A polymorphism (407±69 kPa) is two times higher than the one determined for noncarriers (195.4±48.7 kPa; p<0.05). The background activity of platelets measured as an interrelation of Cd...
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