Respiratory infections are a real threat for humans, and therefore the pig model is of interest for studies. As one of a case for studies, Actinobacillus pleuropneumoniae (APP) caused infections and still worries many pig breeders around the world. To better understand the influence of pathogenic effect of APP on a respiratory system—lungs and tracheobronchial lymph nodes (TBLN), we aimed to employ matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI). In this study, six pigs were intranasally infected by APP and two were used as non-infected control, and 48 cryosections have been obtained. MALDI-TOF MSI and immunohistochemistry (IHC) were used to study spatial distribution of infectious markers, especially interleukins, in cryosections of porcine tissues of lungs (necrotic area, marginal zone) and tracheobronchial lymph nodes (TBLN) from pigs infected by APP. CD163, interleukin 1β (IL‑1β) and a protegrin-4 precursor were successfully detected based on their tryptic fragments. CD163 and IL‑1β were confirmed also by IHC. The protegrin-4 precursor was identified by MALDI-TOF/TOF directly on the tissue cryosections. CD163, IL‑1β and protegrin‑4 precursor were all significantly (p < 0.001) more expressed in necrotic areas of lungs infected by APP than in marginal zone, TBLN and in control lungs.

Background With age, facial muscles lose the ability to complete contractions properly resulting in limitation of facial expressions, fat shifting, and leading to skin creases and wrinkling. Objectives The aim of this study was to determine the effects of the novel high intensity facial electromagnetic stimulation (HIFES) technology combined with synchronized radiofrequency on delicate facial muscles, using an animal porcine model. Methods Eight (n = 8, 60-80 kg) sows were divided into the active group (n = 6) and the control group (n = 2). The active group underwent four 20-minute treatments with radiofrequency (RF) and HIFES energies. The control group was not treated. Histology samples of muscle tissue were collected by a punch biopsy (6 mm in diameter) from the treatment area of each animal at baseline, 1-month, and 2-month follow-up. The evaluation included staining of the obtained tissue slices with hematoxylin and eosin (H&E) and Masson’s Trichrome to determine the changes in muscle mass density, number of myonuclei, and muscle fibers. Results The active group showed muscle mass density increase (by 19.2%, p &lt; 0.001), together with elevated numbers of myonuclei (by 21.2%, p &lt; 0.05) and individual muscle fibers, which increased from 56.8 ± 7.1 to 68.0 ± 8.6 (p &lt; 0.001). In the control group, no significant changes were seen in any of the studied parameters throughout the study (p &gt; 0.05). Finally, no adverse events or side effects were observed in the treated animals. Conclusions The results document favorable changes after the HIFES + RF procedure at the level of the muscle tissue, which may be of great importance in terms of maintenance of facial appearance in human subjects.

Proper histological evaluation of lung tissue and accurate antigen detection by immunodetection techniques require histological tissue processing to preserve tissue reactivity and open alveolar spaces. In this study, we focused on testing and comparing different procedures of tissue cryopreservation. Sucrose or Tissue Tek were used with several methods of freezing samples by supercooled liquids and liquid nitrogen. Changes in tissue caused during the freezing of samples and the effect of cryoprotectants on the tissue were recorded. Rat and porcine pulmonary tissues were used in this experiment. This study aimed to optimize the process of lung cryopreservation with emphasis on enabling proper anatomical evaluation and preserving a high tissue immunoreactivity. The best results were obtained by inflating pulmonary parenchyma with a 1 : 1 mixture of O.C.T. with phosphate buffered saline (PBS) frozen in supercooled n-heptane placed on dry ice. Pulmonary tissue prepared in this way enables to perform proper histological evaluation and to detect target molecules by immunohistochemical analysis.

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