Background: Thermal denaturation experiments were extended to study the thermal behaviour of the main motor proteins (actin and myosin) in their native environment in striated muscle fibres. The interaction of actin with myosin in the highly organized muscle structure is affected by internal forces; therefore their altered conformation and interaction may differ from those obtained in solution. The energetics of long functioning intermediate states of ATP hydrolysis cycle was studied in muscle fibres by differential scanning calorimetry (DSC). Results: SETARAM Micro DSC-II was used to monitor the thermal denaturation of the fibre system in rigor and in the presence of nucleotide and nucleotide analogues. The AM.ADP.P_i state of the ATP hydrolysis cycle has a very short lifetime therefore, we mimicked the different intermediate states with AMP.PNP and/or inorganic phosphate analogues V_i and AlF₄ or BeF_x. Studying glycerol-extracted muscle fibres from the rabbit psoas muscle by DSC, three characteristic thermal transitions were detected in rigor. The thermal transitions can be assigned to myosin heads, myosin rods and actin with transition temperatures (T_m) of 52.9 ± 0.7°C, 57.9 ± 0.7°C, 63.7 ± 1.0°C. In different intermediate states of the ATP hydrolysis mimicked by nucleotide analogues a fourth thermal transition was also detected which is very likely connected with nucleotide binding domain of myosin and/or actin filaments. This transition temperature T_m4 depended on the mimicked intermediate states, and varied in the range of 66°C – 77°C. Conclusion: According to DSC measurements, strongly and weakly binding states of myosin to actin were significantly different. In the presence of ADP only a moderate change of the DSC pattern was detected in comparison with rigor, whereas in ADP.P_i state trapped by V_i, AlF₄ or BeF_x a remarkable stabilization was detected on the myosin head and actin filament which is reflected in a 3.0 – 10.0°C shift in T_m to higher temperature. A similar effect was observed in the case of the nonhydrolyzable AMP.PNP analogue. Differential DSC measurements suggest that stabilization actin structure in the intermediate states of ATP hydrolysis may play an additional role in actin-myosin interaction.

Identification of HPV infection is usually performed on cytological specimens, despite the often transient virus types. HPV profile analysis of pathologically confirmed lesions can also be performed on formalin-fixed paraffin-embedded (FFPE) cone samples and should be taken as standard during follow-up. We compared HPV profiles of cytological and FFPE specimens of women diagnosed with HSIL. Archived PAP smears and FFPE cones from 49 patients were processed. For genotyping, the HPV Direct Flow CHIP test was used. All samples were positive. HPV profile agreement of the two sample types was 84.16–100%. Mono-infections occurred in 12.24% and 61.22% in PAP smears and FFPE specimens, respectively; while multi-infections were detected in 87.76% and 38.78%, respectively. The most abundant genotypes were HPVs 16, 31, and 51/33. Of all infections, 56.25% and 64.93% were caused by nonavalent vaccinated type (VT) HPVs; while 50.69% and 38.96% belonged to non-nonavalent VT HPVs, in PAP smears and FFPE specimens, respectively. Our results confirmed the importance of HPV genotyping of FFPE cone samples. We also confirmed a remarkable presence of non-vaccinated HPV types in HSIL cases indicating the importance of vaccine development.

The association of clinical variables with body mass index (BMI) and changes experienced during a gluten-free diet (GFD) in celiac disease (CD) is not well established. In this retrospective cohort study, we aimed to investigate factors aligned with baseline and a follow-up regarding BMI in CD cases diagnosed at the University of Pécs (Hungary). Data were collected regarding gender, age, clinical presentation, histology, serology, extraintestinal manifestations, and BMI upon diagnosis and during follow-up. To compare variables with baseline BMI and BMI changes in short-, intermediate-, and long-term periods, we applied univariate analyses. A total of 192 CD patients were included. Males had significantly higher mean BMI when compared with females at diagnosis (22.9 ± 4.1 vs. 21.4 ± 4.3 kg/m², p = 0.041) and during follow-up (p = 0.031, p = 0.029, and p = 0.033 for short-, intermediate-, and long-term follow-ups, respectively). Non-classical CD patients experienced higher mean BMI at diagnosis (22.9 ± 4.0 vs. 20.7 ± 4.4 kg/m², p < 0.001) and following long-term follow-up (24.5 ± 3.2 vs. 22.6 ± 3.4 kg/m², p = 0.039) than classical patients. In conclusion, although the mean BMI remained in the normal range, it increased significantly during follow-up, even at the short-term follow-up. This change was characteristic for non-classical cases and males on the long-term follow-ups.

Polycyclic aromatic hydrocarbons (PAHs) can be extremely hazardous to human health due to their carcinogenicity and toxicity. Following mother exposure, a part of PAHs might be transferred into breast milk, and PAH exposure may results in adverse effects in breastfeeding infants. Chrysene, benzo(a,h)anthracene, benzo(b)fluoranthene and benzo(a)pyrene (PAH4) are the most suitable indicators of PAHs in food. The aim of this work was to develop a sensitive and selective sample preparation method for the determination of PAH4 in breast milk using gas chromatography-mass spectrometry. The study compared different extraction methods of PAHs. The method was validated using parameters such as linearity, precision, accuracy, limit of detection (LOD) and limit of quantification (LOQ). The calibration curve was linear over the range of 0.5–50 ng/mL. The intra- and inter-day relative standard deviation values were less than 0.37%. The recovery values ranged between 99.97 and 99.99%. The LOD and LOQ values were found to be between 1.4 and 1.7 ng/mL and between 2.5 and 4 ng/mL, respectively.