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Journal of Thermal Analysis and Calorimetry

, Volume 133, Issue 1, pp 579–589 | Cite as

New possibilities of application of DSC as a new clinical diagnostic method

A review
  • Péter Farkas
  • Franciska Könczöl
  • Dénes Lőrinczy
Article
  • 163 Downloads

Abstract

Differential scanning calorimetry (DSC) is the most often used method in thermal analysis. Recently, there is growing clinical use for it. With this method, we can measure the effects of drugs and we can quantify and characterize changes in different organs or parts of body. This way it is possible to conclude short- and long-term effects in a predictive way. Our team were examined the effects of cyclophosphamide on guinea pigs equivalent to human protocol in experimental conditions. Besides of its beneficial effects, cyclophosphamides may have got severe life-threatening side effects and complications because of the actual plasma level and high cumulative dosage. In the first step of our experiment, we examined the effects of cyclophosphamide on guinea pigs’ nerve–muscle complex with oncological indication by using a dosage protocol based on body mass. As a second step with the same method and cyclophosphamide dosage, we examined its effect on the heart left ventricle. The third step was in the further clinical application experiment with unchanged parameters on blood plasma as well as blood cells too, exhibiting dosage-dependent changes on plasma and blood cells. Alterations in different materials caused by using different cyclophosphamide dosage and not uniform treatment length produced well correlated and clearly detected changes with DSC. Based on our results, we found detectable and partway quantified alterations with DSC on blood plasma components; therefore, it can be used in clinical routine because it is relatively simple and cheap. In long-term treatments, incidental severe results and side effects caused by cumulative dose may become also predictive with this method. All these show a new promising area in DSC usage which passed out of mind in the last 10 years.

Keywords

Cyclophosphamide Nerve Muscle Blood plasma and cell DSC 

Introduction

Differential scanning calorimetry (DSC) is the most often used method in thermal analysis. Recently, the clinical use is growing for it. With this method, we can measure the effects of drugs, free radicals and poisons [1, 2, 3, 4, 5, 6, 7, 8], work-related lesions [9, 10] and/or we can quantify and characterize changes in organs or different parts of body caused by or different diseases [11, 12, 13, 14, 15, 16, 17, 18, 19] or medical interventions [20, 21]. This way it is possible to conclude short- and long-term effects in a predictive way. Our team were examined the effects of cyclophosphamide on guinea pigs equivalent to human protocol in experimental conditions. Cyclophosphamide is a very effective immunosuppressive drug, and it is used primarily in severe immunological diseases associated with organ involvement [22]. The metabolism of cyclophosphamide is partly genetically determined and partly influenced by comorbidities and incidental treatment. In case of tumors and well-known autoimmune diseases with different pharmacokinetics, it shows a large individual variability and may change with time. Besides of its beneficial effects, cyclophosphamides may have got severe life-threatening side effects and complications because of the actual plasma level and high cumulative dosage [23, 24].

In the first step of our experiments, we examined the effects of cyclophosphamide on guinea pigs’ nerve–muscle complex with oncological indication by using a dosage protocol based on body mass [25, 26]. The analysis was made by a SETARAM Micro DSC-II calorimeter. According to our results, we could show a dose-dependent difference between thermal parameters of untreated and treated samples which proved that cyclophosphamide has got a peripheral and in smaller degree of cases muscle damage effect. All of that it showed the effect of cyclophosphamide like this also verified the method’s efficiency in detecting changes.

As a second step with the same method and cyclophosphamide dosage, we examined its effect on the heart muscle [27]. The reason of it was that alkylating agents such as cyclophosphamide (di-(beta-chloroethyl) aminopropyl-phosphor acid–ester–amide) has got severe, fatal cardiac damage effect. It was discovered more than 40 years ago apropos of bonemarrow transplantation [28, 29]. The cyclophosphamide treatment causes acute cardiac failure or death only in a few cases and has not got cumulative nature contrary with other drugs [30]. The cyclophosphamide induced cardiomyopathy based on the accumulation of free radicals and the decreasing number of antioxidants [31]. Metabolites of cyclophosphamide—which are made by monooxigenase enzyme (cytochrome P450) of liver—cause oxidative stress and direct capillary endothelial damage resulting in the extravasation of proteins/red blood cells and toxic metabolites that damage cardiac muscle. In this case, micro-thrombus, interstitial hemorrhages and edema will emerge in the myocardium and during autopsy we can find capillary micro-thrombus and fibrin deposition in the enlarged interstitium which is specific for cyclophosphamide-induced cardiomyopathy [32]. In clinical routine symptoms appear within 48 h after drug administration or in the first 10 days, these are tachyarrhythmia, hypotension, cardiac failure, myocarditis, pericarditis and pericardial tamponade [23]. Hemorrhagic myocarditis is rare usually quick and fatal [32]. High dose of drugs increases the appearance of this condition, but limit values are not clarified yet. Cardiac failure may be regarded as risk factor [24]. There had been made attempts to detect early changes in cardiac muscle using MRI but clear predictive changes could not been identified [33].

The third step was in the further clinical application experiment with unchanged parameters on blood plasma as well as blood cells too, exhibiting dosage-dependent changes on plasma and blood cells [34]. Collectively evaluating the performed studies, a clear correlation can be observed in the results obtained on different experimental materials. Alterations in different materials caused by using different cyclophosphamide dosage and not uniform treatment length produced well correlated and clearly detected changes with DSC. If we can detect the drug-induced changes for example in blood plasma, we can conclude effects in other areas too. This technique has proven its applicability in case of different malignant diseases too, giving results with diagnostic value [35, 36, 37, 38, 39, 40, 41, 42]. Based on our results, we found detectable and partway quantified alterations with DSC on blood plasma components; therefore, it can be used in clinical routine [11, 12, 13, 14, 15, 16, 17, 18, 19]. The testing of blood plasma with DSC is relatively simple and cheap. If we could define quantifiable relationships between denaturation parameters and modifications in curves, then we can get information about induced effects in other areas of the body. In long-term treatments, incidental severe results and side effects caused by cumulative dose may become predictive with this method. Taking a blood sample is minimally invasive, therefore acceptable for the patients. The evaluation of sample could be partially automated, so we can get a relative fast result and make further therapy decision. If we can manage to predict the harmful effects for patients which are arising from different factors, we could prevent them with this method by decreasing the dose or changing to other drug. This is beneficial for the patient and treatment too. All these show a new promising area in DSC usage which passed out of mind in the last 10 years. To that we could determine the exact drug dosage with this method, and it is necessary to safely quantify the detectable changes by considering the modifications caused by disease. To do this, it is necessary to specify the curve and parameter changes [43, 44], but extensive patient examinations need to be performed too.

Effect of cyclophosphamide on muscle–nerve samples

The treated animals (all together 60 guinea pigs, Cavia porcellus) were distributed in seven groups in cases of nerves and muscles: control and treated with 1–6 injections (see Table 1, for both samples). The preparation of samples, the chemotherapy as well as the DSC experiments were performed as it was described in [25, 26], having a valid ethical permission (BA02/2000-4/2012). The dose-dependent effect of cyclophosphamide treatment can already be seen in Fig. 1 too. The shift of main denaturation temperatures into smaller range, the decrease in maximum heat flow can it demonstrate. The most striking effect of the drugs was a definite alteration in thermal parameters at the left side of the animals (see Table 1). One obvious explanation could be the right-handed assistant (to held the animal on its back during the injection) and the right-handed medical doctor (who made the treatment), because this way the drug was administrated into the left side of the animal, thereby the medicine had longer time to impact its effect at this side.
Table 1

The characteristic thermal parameters of the denaturation after the cyclophosphamide treatment of guinea pig sciatic nerve samples

Treatment of sciatic nerve

Thermal parameters

Right

Left

T m1 /°C

T m2/°C

T m3/°C

ΔH c/J g−1

T m1/°C

T m2/°C

T m3/°C

ΔH c/J g−1

Control (5)

53.4

61.2

63.3

5.5

54.7

61.4

64.2

5.9

One injection (5)

 1i-E

53.4

61.7

65.4

3.9

53.4

61.0

66.3

3.4

 1i-3d-E

53.0

59.9

62.7

3.5

52.9

59.9

63.6

3.2

 1i-5d-E

54.5

61.1

65.5

3.9

54.7

62.0

67.1

3.6

Two injections (5)

 2i-E

54.1

61.3

65.0

4.1

53.7

61.3

65.0

4.3

 2i-3d-E

53.7

61.4

65.1

3.5

53.8

61.2

65.2

3.9

 2i-5d-E

53.0

60.8

65.3

3.4

53

61.1

72.0

3.9

 1i-3d-1i-3d-E

54.2

60.4

64.5

2.1

53.4

61.2

64.9

2.4

Three injections (5)

 2i-5d-1i- 3d-E

53.8

59.6

64.0

5.3

53.9

60.2

64.5

4.7

Four injections (5)

 2i-5d-1i-3d-1i-E)

53.7

60.0

65.0

4.0

53.9

60.2

65.0

4.3

Five injections (5)

 2i-5d-1i-3d-2i-E

53.0

60.7

64.6

3.32

55.0

60.7

64.8

3.0

Six injections (5-2E)

 2i-5d-1i-3d-3i-E

52.4

61.8

64.6

5.4

53.4

61.5

65

3.5

Data are averages, and rounded to one decimal place

T m melting temperature, ΔH c calorimetric enthalpy, i injection, d days between treatments, E exit

Italic: data out of ± 2× s.d., bold: data out of 3× s.d., n = 5

Fig. 1

Dose-dependent DSC scans of sciatic nerve in case of cyclophosphamide-treated Cavia porcellus (determination of T ms of scans was made by deconvolution, which is not shown in Figs. 1, 2, 46)

In case of right-sided nerves, the thermal domain characterized by T m3, and the total calorimetric enthalpy can monitor the dose dependence of the treatment while in T m1 and T m2 we could observe only a very mild tendency (it depended on number of injection and time elapsed till the exit up to two treatments. T m1 showed a dose dependence only after six injections and in the treatment 1i-5d-E). In contrast with these data in the left side, the thermal stability of domains characterized by T m1 and T m3 exhibited strong dose dependence up to two injections. In case of greater doses, T m1 and T m2 thermal units are more sensitive, while the total calorimetric enthalpy differs from control one practically in all cases (it lies out of average ± 3× s.d. of the control). We have no any information about the structural and functional role of those domains which produced these thermal events.

We were in a better position to interpret the possible structural damage in m. gastrocnemii, because we have enough experience in the thermal denaturation of skeletal muscles either from basic research or in clinical point of view [6, 7, 8, 9, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56]. The denaturation heat flow in these cases can be separated into definite three big parts (see Fig. 2). On the basis of our previous works [45, 46, 47, 48, 49, 50], the middle part is the rod part of myosin (it is the less sensitive for the different kinds of influences), the lower denaturation peak could be assigned to the myosin head (which is an ATPase enzyme, its damage abruptly appears in the function too), while the highest melting is actin plus actin–myosin interaction (actomyosin complex, see Fig. 3). The actual molecular dynamic state of actin, which is a very sensitive monitor of any kind of attack against its nucleotide binding domain [57, 58, 59, 60, 61, 62], will modify the whole contraction cycle. The dose dependence of drug treatment can be followed in Fig. 2. The thermal parameters of heat denaturation are in Table 2. Similar side effect can be observed as in case of n. ischiadicus. The change in T ms and ΔH is similar than in case of nerves. Structurally it means that the myosin head (muscle “motor,” the chemical energy transformation into mechanical one) and actin filaments (molecular dynamic feature, that is the base of locomotion) are mainly affected (T m1 and T m3), which physiologically appears as worse actomyosin complex function, finally in the bad locomotion capability. In case of dose of six injections, we had two exits—with unknown reason—before the end of experiments (after the fifth injection).
Fig. 2

Thermal denaturation of drug-treated m. gastrocnemii

Fig. 3

Deconvolution of denaturation curve in case of rabbit psoas muscle to demonstrate the possible contribution of different protein site in the denaturation process (see, e.g. [44, 48])

Table 2

The characteristic thermal parameters of the denaturation after the cyclophosphamide treatment of guinea pig m. gastrocnemius samples

Treatment of m. gastrocnemius

Thermal parameters

Right

Left

T m1/°C

T m2/°C

T m3/°C

ΔH c /J g−1

T m1/°C

T m2/°C

T m3/°C

ΔH c /J g−1

Control (5)

50.2

57.0

67.3

2.9

49.7

56.8

68.1

3.2

One injection (5)

 1i-E

50.1

56.8

66

1.9

50.2

56.9

64.9

2.2

 1i-3d-E

49.5

56.6

66.9

2.3

49.8

56.6

66.8

2.3

 1i-5d-E

50.0

56.8

66.5

2.5

50.3

56.9

66.7

2.2

Two injections (5)

 2i-E

49.9

56.7

67.8

2.7

49.9

56.9

67.3

2.5

 2i-3d-E

50.2

56.6

66.7

2.4

50.3

56.5

66.4

2.3

 2i-5d-E

49.7

56.9

67.4

2.6

50.2

56.7

67.6

2.8

 1i-3d-1i-3d-E

49.9

56.6

65.6

2.3

49.9

56.7

67.5

2.6

Three injections (5)

 2i-5d-1i-3d-E

50.9

56.5

64.4

2.9

51.0

56.5

64.7

2.7

Four injections (5)

 2i-5d-1i-3d-1i-E)

50.0

56.4

66.6

2.6

50.4

56.7

66.2

2.5

Five injections (5)

 2i-5d-1i-3d-2i-E

50.0

56.10

68.0

3.8

50.0

56.4

66.3

3.4

Six injections (5-2E)

 2i-5d-1i-3d-3i-E

50.4

56.4

65.8

2.2

50.4

56.5

65.4

2.4

Data are averages, and rounded to one decimal place

T m melting temperature, ΔH c calorimetric enthalpy, i injection, d days between treatments, E exit

Italic: data out of ± 2× s.d., bold: data out of ± 3× s.d., n = 5

Consequences of chemotherapeutic treatment in the left ventricle of the heart

The dose-dependent effect of cyclophosphamide treatment on left ventricle of guinea pig’s heart can be seen in Fig. 4, where the shifts in melting temperatures and the change of calorimetric enthalpy clearly demonstrate it. From structural and functional point of view, the heart muscle basically is similar to the skeletal muscle, this way during the interpretation of DSC scans we can use the information obtained at skeletal muscle. The denaturation heat flow can also be divided into three big parts. On the basis of our previous works [45, 46, 47, 48, 49, 50], the melting around 58 °C is the rod part of myosin, the lower denaturation peak could be assigned to the myosin head, while the highest melting above 62 °C is actin plus actin–myosin interaction (actomyosin complex, and the side effect of the drug bound to these proteins). The dose dependence of drug treatment can be followed in Fig. 4. The thermal parameters of heat denaturation are in Table 3. Up to two injections, T m1, T m3 and the calorimetric enthalpies are influenced, that is the myosin head and actin are more sensitive for the treatment. In case of three and more cyclophosphamide treatment, T m2, T m3 and the calorimetric enthalpies can monitor the effect, that is the myosin rod and actin are mainly attacked. In our institute were partly performed those experiments [59, 60, 61, 62], which demonstrated how can influence the molecular dynamic characteristic of actin by the manipulation of their structure with different nucleotides. The cyclophosphamide is a relatively small molecule, so we can assume that it can bound easy or to the nucleotide binding place of actin or on the surface of the filament. Taking the whole procedure, we can say that the myosin head and actin filaments are mainly affected (T m1 and T m3), which physiologically appears as worse actomyosin complex function, finally in the bad pulmonary capability. This way the treatment can cause severe problem in the function of the heart and circulation. In case of dose of six injections, we had two exits before the end of experiments (after the fifth injection).
Fig. 4

Curves of heart muscle (left ventricle) during denaturation

Table 3

The characteristic thermal parameters of the denaturation after the cyclophosphamide treatment of guinea pig left ventricle samples

 

T m1/°C

T m2/°C

T m3/°C

ΔH c/J g−1

Control (5)

56.1

59.0

62.0

2.1

One injection (5)

 1i-E

56.0

56.8

60.8

1.4

 1i-3d-E

55.5

59.0

63.0

1.6

 1i-5d-E

55.9

58.7

61.6

1.7

Two injections (5)

 2i-E

56.4

59.1

61.8

1.7

 2i-3d-E

56.6

59.2

62.0

2.0

 2i-5d-E

56.2

59.1

61.5

1.8

 1i-3d-1i-3d-E

55.2

59.0

63.1

1.8

Three injections (5)

 2i-5d-1i-3d-E

55.8

58.3

61.3

1.6

Four injections (5)

 2i-5d-1i-3d-1i-E

49.9

55.9

60.6

1.8

Five injections (5)

 2i-5d-1i-3d-2i-E

55.7

58.6

61.6

1.4

Six injections (5-2E)

 2i-5d-1i-3d-3i-E

55.8

58.7

61.4

1.7

T m melting temperature, ΔH c calorimetric enthalpy normalized on sample mass, data are averages, and rounded to one decimal place

Italic: data within ± 2× s.d., bold: data within ± 3× s.d., n = 5

Red blood cells and blood plasma as a damage monitoring of treated samples

In our blood study, we examined red blood cells beside plasma too. The largest proportions of blood cells are red blood cells, and they are responsible for gas transport and acid–base balance in the body [63]. The early calorimetric investigations of erythrocytes were made by Monti and Wadsö, comparing the heat production of normal and anemic states [64]. It was followed by checking the treatment in hyperthyroid case [65], investigating the effect of methylene blue stimulation [66], the pH, temperature, glucose concentration and storage conditions [67]. They made the first comparison—using red blood cells—between the different calorimetric techniques, preparation techniques and suspension media [68]. Hernández et al. [69] made an interesting study referring to the alterations in erythrocyte membrane protein composition in advanced non-small cell lung tumor.

According to the current knowledge, cyclophosphamide is transported in plasma in dissolved form and a small number of cases (~ 20%) binding to plasma proteins, in this process red blood cells do not participate [70]. One of the cyclophosphamide well-known side effects is transitional white blood cell and red blood cell reduction caused by its cytotoxic effect. Due to this, we expect deviation compared to control on the curves of red blood cells. In case of red blood cell mixture (preparation is described in [34]), the most interesting finding was a four-step transition in case of control (see Fig. 5): a low-temperature melting around 57 °C (it is extremely small, we did not involve it into Table 4), as well as endotherms around 69–76–84 °C. These transitions are shifting to lower level with increased number of treatment and from the fifth injection practically remaining only one endotherm. This finding can play an important role to plan the medical interventions and the tight controls in case of human applications. In the first group (one injection and next day termination), the higher three endotherms decreased remarkably. In groups handled with two injections, only T m3 decreased definitely (in T m1 and T m2 there were fluctuations). In groups where three and four injections were administrated and all melting temperatures decreased. The injected cyclophosphamide is partly connecting to the blood cells and partly making some alterations on them.
Fig. 5

Thermal responses of red blood cells during denaturation

Table 4

The characteristic thermal parameters of the denaturation of guinea pig’s blood plasma and red blood cell samples in the function of the cyclophosphamide treatment

Treatments

Thermal parameters

Plasma

Red blood cell

T m1/°C

T m2/°C

T m3/°C

ΔH c /J g−1

T m1/°C

T m2/°C

T m3/°C

ΔH c /J g−1

Control (5)

59.9

62.3

84.7

0.8

68.7

75.3

84.5

4.7

One injection (5)

 1i-E

62.5

87.5

92.3

0.8

53.8

68.9

74.2

5.2

 1i-3d-E

58.1

66.9

79.3

0.5

67.9

73.4

84.5

5.4

 1i-5d-E

62.4

83.4

91.4

0.8

68.4

75.9

84.0

5.5

Two injections (5)

 2i-E

58.7

65.4

79.4

0.7

68.5

74.7

82.5

4.9

 2i-3d-E

60.5

66.2

81.6

0.6

66.5

73.1

81.3

4.7

 2i-5d-E

60.0

64.4

82.2

0.5

68.5

74.8

81.6

4.6

 1i-3d-1i-3d-E

55.8

66.0

80.1

0.5

68.3

72.6

79.9

4.2

Three injections (5)

 2i-5d-1i-3d-E

55.4

62.0

67.3

0.60

67.3

73.3

79.8

4.0

Four injections (5)

 2i-5d-1i-3d-1i-E)

59.8

65.5

72.0

0.3

70.8

72.5

83.5

5.5

Five injections (5)

 2i-5d-1i-3d-2i-E

57.9

68.7

72.8

0.2

71.8

4.2

Six injections (5-2*)

 2i-5d-1i-3d-3i-E

55.7

59.7

64.8

0.7

68.6

74.9

3.4

Data are averages, and rounded to one decimal place

T m melting temperature, ΔH c calorimetric enthalpy, normalized on sample mass)

Italic: within ± 2× s.d., bold: significant difference at p < 0.05, n = 5

* significant difference compared to the control

Plasma is total up to 25% of the extracellular fluid space—“milieu intérieur”—this is a moving compartment and creates relationships between organs and body systems. Plasma consists of 90% of water, diffusible compartments and non-diffusible plasma proteins. Using electrophoresis in plasma proteins, we can distinguish six main fractions. These are albumin, α 1-, α 2-, ß- and γ-globulin as well as fibrinogen, but in fact we know more than 200 proteins with function. The most important transport proteins are for example transferrin, ceruloplasmin, thyroxine-binding globulin, transcortin, transcobalamin and haptoglobin. Proteins have an effect on fluid distribution in body, acid–base balance and immune function. Substances given into blood path such as intravenous drugs distribute uniformly in intravascular space in 5 min [63].

The dose-dependent effect of cyclophosphamide treatment on plasma can be seen in Fig. 6. The shift of melting temperatures into higher range (except of T m1), the decrease in maximum heat flow can it demonstrate (for the sake of showing the trend in the severity of the treatment we plotted only the effect of 1–3–5 treatments). In our experiments, we can generally suppose the present at least three different main thermal structural units in all cases. The more effected compounds are the transitions characterized by T m2 and T m3 (see Table 4). The start of the treatment shows severe change in T ms (definite increase) and the effect of the time of exit (in case one injection). The calorimetric enthalpy decreases remarkably only from the second intervention. From the third injection appears a big decrease in all thermal parameter.
Fig. 6

Denaturation curves of blood plasma as the most sensitive biological sample for the chemotherapy

During the treatment of any disease, the cyclophosphamide plasma level has got an important role in expected therapeutic effect and side effects and complications too. The metabolism of cyclophosphamide may show large-scale individual variability which is partly genetically determined and partly under the influence of co-diseases or other associated treatments that’s why it may changes over time [70]. Joy et al. showed that cyclophosphamide’s pharmacokinetics and 4-hydroxy-cyclophosphamide diverge in case of autoimmune diseases (glomerulonephritis) and cancer. In their study, they used liquid chromatography—mass spectroscopy to detect the cyclophosphamide level of plasma which is an effective but circumstantial method [71].

The first exciting studies on thermal behavior of human plasma in case of patients sick in cancer were made by Monaselidze and his colleagues [72, 73]. It was followed by a comparative DSC study of human and bovine serum albumin [74]. In the last years, numerous studies had been published for plasma examination with DSC and they showed that the received curve is very sensitive and informative in detecting some types of changes [35, 36, 37, 38, 39, 40, 41]. Diagnostic effectiveness of this method has been confirmed among others in psoriasis, chronic obstructive pulmonary disease (COPD), breast cancer, melanoma, multiple myeloma [37] and schizophrenia [75]. Testing the main proteins stability of blood plasma in case of patients with ductal carcinoma in post-surgery period seems to be a good indicator of the success of surgical intervention [76]. Recently DSC has been employed for diagnostics and characterization of the changes in the brain at molecular and supramolecular level associated with drug-induced neuro-degenerative disorders. In order to test the DSC potential, Tonchev and his colleagues used an experimental animal model (mouse) of scopolamine-induced dementia of Alzheimer’s disease (AD) type [77, 78]. The biggest success in the line of application DSC as a diagnostic tool in clinical practice was achieved by Garbett and her group [43, 79]. They have developed a special algorithm to make some special “vectors” from DSC data, and with the aid of this procedure in case of prediction Lupus (a severe immunological disease, its diagnose is very difficult) they accomplished 82% success in the prediction of this illness (they had 300 control and 300 patients!).

Conclusions

In our study, we examined the cyclophosphamide caused effects on mainly attacked tissues, body compounds in our model. We have presented data on strong dose-dependent effect in case of sciatic nerve (due to the lag of histological result without the identification of possible target points). In case of muscle cells (skeletal or heart origin), the myosin head and the actin filaments exhibited the biggest answer. The possible explanation is given in the proper paragraph of this review, and now we mention some additional results. The DSC can be efficiently used in the characterization of actin under the different conditions. The actin-binding proteins (cofilin, twinfilin and toxofilin) can effectively influence the thermal stability of actin [80, 81, 82, 83] (very probably the cyclophosphamide too), which could explain the changes in T m3 and ∆H c (see Tables 2, 3). Similar result is that filaments prepared from ADP-α-cardiac actin monomers were thermodynamically less stable compared to the ADP-α-skeletal actin protomers containing filaments [84] (this could explain the difference between heart and skeletal muscle T m3 and ∆H c too).

The blood plasma and red blood cells were the most sensitive samples reflecting on chemotherapy treatment. The detected changes may partly correlate with the concentration of cyclophosphamide and the drug-induced biological effects in plasma components and red blood cells. Based on biological regularity, these effects most likely cause functional consequences. These changes in side effects supposed to have an important role; therefore, detecting the degree of the alteration’s severity and course may be predictive.

Considering that relative fast and informative determination of plasma level of cyclophosphamide is slightly difficult in clinical routine, the application of this method (DSC) in this area is promising. Nevertheless, the accurate prognosis of cyclophosphamide’s side effects has got an important role and may correlate with the degree and quality of induced effects on blood components. In order to clarify these results, further studies may be needed with increased number of samples to make statistically well-based conclusions, especially because the realization of our method (DSC) in clinical application is in progress (giving, e.g., 82% precision in the diagnosis of Lupus [79]) and the number of publications in this field are growing.

Notes

Acknowledgements

This work was supported by Grant OTKA CO-272 (for D. Lőrinczy). The present scientific contribution is dedicated to the 650th anniversary of the foundation of the University of Pécs, Hungary.

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Copyright information

© Akadémiai Kiadó, Budapest, Hungary 2017

Authors and Affiliations

  • Péter Farkas
    • 1
  • Franciska Könczöl
    • 2
  • Dénes Lőrinczy
    • 3
  1. 1.Clinics of Radiology Clinical CenterUniversity PécsPécsHungary
  2. 2.Institute of Forensic Medicine, University PécsPécsHungary
  3. 3.Institute of Biophysics School of Medicine, University PécsPécsHungary

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