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Background With age, facial muscles lose the ability to complete contractions properly, resulting in limitation of facial expressions and 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 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 < .001), together with elevated numbers of myonuclei (by 21.2%, P < .05) and individual muscle fibers, which increased from 56.8 ± 7.1 to 68.0 ± 8.6 (P < .001). In the control group, no significant changes were seen in any of the studied parameters throughout the study (P > .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 patients.

Background Multiple studies have reported adipose tissue reduction after the application of the High‐Intensity Focused Electromagnetic (HIFEM) field technology, yet cellular level evidence of the mechanisms has remained scarce. Objectives This study aims to verify or refute previous single‐study histological evidence and further investigates the proposed mechanism of apoptotic induction. Methods The thigh of two Large White pigs was treated with HIFEM for 30 minutes. Fat punch biopsies were collected from the application area before, immediately after, and 8 hours post‐treatment. Control samples were taken from the abdomen immediately after and 8 hours post‐treatment. Samples were analyzed for pro‐apoptotic DNA markers (BAX, BCL‐2, TXNIP, MMP9, TNF‐α), the levels of free fatty acids (FFA), and the pH levels of the adipose tissue. Results The levels of FFA in the treated adipose tissue increased on average by 127.1% immediately post‐treatment and by 134.1% 8 hours post‐treatment, indicating a rapid breakdown of lipids. The average recorded adipose pH changed from 7.30 ± 0.12 at baseline to 6.60 ± 0.07 immediately post‐treatment (P = .001) and to 7.19 ± 0.12 8 hours post‐treatment. The levels of BAX, TXNIP, MMP9, and TNF‐α increased post‐treatment while BCL‐2 decreased. Control samples showed constant levels of pH and pro‐apoptotic markers. The FFAs in the control samples were increased by 41.6%‐51.4%. Conclusion The changes in the levels of the pro‐apoptotic markers conformed to the previously reported elevated fat apoptosis post‐HIFEM treatments. These effects were accompanied by an increase in FFA levels, and by reduced pH levels, due to the increased acidity in the adipose tissue. Further research is required to explore the potential of nonthermal induction of apoptosis.

Streptococcus suis represents a primary health problem (such as meningitis, septicemia and arthritis in piglets and fatteners) in the swine industry worldwide and also an emerging zoonotic pathogen. In the Czech Republic, many pig farms repopulated their herds over the past decades to reduce morbidity and minimize treatment. The study analysed serotypes, sequence types and antimicrobial susceptibility in 39 S. suis isolates obtained from organs of diseased pigs from selected 16 repopulated farms with a history of S. suis-associated diseases and routine antimicrobial treatment with tulathromycin and/or amoxicillin. The analysis revealed diversity of collected isolates with regular occurrence of more than three serotypes per farm. The serotypes identified were 1/2 and 7, each in six isolates, followed by serotype 2 and 3 found in five isolates each, other serotypes were less frequent. Seven isolates were not typable by multiplex PCR and we also found sequence type of unknown type in thirteen isolates. The majority of S. suis isolates were resistant to clindamycin (n = 31), tetracycline (n = 29) and tilmicosin and tulathromycin (n = 28). On the other hand, with the exception of two isolates that were intermediately susceptible to penicillin and ampicillin, all isolates were susceptible to all three tested subgroups of beta-lactam antibiotics.

BACKGROUND Radiofrequency (RF) and high-intensity focused electromagnetic (HIFEM) technologies are used for noninvasive body shaping as standalone modalities. OBJECTIVE To examine the effects of novel synchronized RF and HIFEM on subcutaneous adipose tissue in a porcine animal model. MATERIALS AND METHODS Seven large white pigs aged 6 months received 3 abdominal treatments of simultaneous application of synchronized RF and HIFEM (30 minutes, once per week). Punch biopsies of treated and control subcutaneous tissue were collected at the baseline, 4 days, 2 weeks, 1 month, and 2 months. Specimens were examined by light and scanning electron microscopy. Adipocyte volume was analyzed. Fat tissue temperature was measured in situ (fiber optic probes) and superficially (thermal imager). RESULTS Fat layer was heated to temperatures of 42 to 45°C. Signs of fat apoptosis (shape alternations and pyknotic nuclei) appeared at day 4 and peaked between 2 weeks and 1 month. Adipocyte volume decreased significantly (p < .001) by 31.1% at 2 weeks, 1 month (-23.6%), and 2 months (-22.0%). Control samples showed healthy adipocytes. Scanning electron microscopy micrographs corroborated histology findings, showing flattened, volume-depleted and disrupted adipocytes. CONCLUSION Synchronized RF with HIFEM procedure resulted in a significant and sustained fat reduction with no adverse events.

Background Myosatellite cells are myogenic stem cells that can transform to provide nuclei for existing muscles or generate new muscle fibers as documented after extended exercise programs. Objectives The authors investigated whether the simultaneous application of High-Intensity Focused Electromagnetic (HIFEM) and Synchrode radiofrequency (RF) affects the levels of satellite cells similarly as the prolonged exercise does to achieve muscle growth. Methods Three 30-minute simultaneous HIFEM and Synchrode RF treatments (once a week) were administered over the abdominal area of 5 Large White swine aged approximately 6 months. All animals were anesthetized during the treatments and biopsy acquisition. Biopsies of muscle tissue were collected at baseline, 4 days, 2 weeks, and 1 month post-treatment. After binding the specific antibodies, the NCAM/CD56 levels, a marker of activated satellite cells, were quantified employing the immunofluorescence microscopy technique with a UV lamp. Results Examined slices showed a continuous increase in satellite cell levels throughout the study. Four days after the treatment, we observed a 26.1% increase in satellite cells, which increased to 30.2% at 2-week follow-up. Additional histological analysis revealed an increase in the cross-sectional area of muscle fibers and the signs of newly formed fibers of small diameters at 2 weeks after the treatment. No damage to muscle tissue and no adverse effects related to the treatment were observed. Conclusions The findings indicate that the simultaneous application of HIFEM and novel Synchrode RF treatment can initiate differentiation of satellite cells to support the growth of existing muscles and, presumably, even the formation of new myofibers.

Structural changes in collagen and elastin fiber density have been previously evaluated by qualitative histological studies; however, quantitative evaluations are lacking.To evaluate quantitative changes in collagen and elastin fibers in the vaginal wall in a porcine model after volumetric radiofrequency heating with an intravaginal applicator.An animal model was used (domestic pig, multipara: 5.67 ± 0.94 deliveries, 3 years of age). Three pigs under general anesthesia were treated (8-minute, vaginal canal area) once per week for the course of three weeks. There were 2 follow-up evaluations at one and four weeks. Histology specimens were obtained via punch biopsy under ultrasound control. Ultrasound video measurements of the vaginal wall thickness were also obtained. Tissue samples were stained by HE as well as stains for collagen and elastin fibers.Elastin (P .001) and collagen (P .01) fiber density increased after every treatment. The measured increase in fibers was highest at the one-week follow-up. Elastin accounted on average for 51.46 ± 16.86% of the tissue examined (increase of 36.8% points), while collagen accounted on average for 44.83 ± 18.92% (increase of 17.1% points). The number of synthetically active cells was increased by 16%. While vaginal wall thickness did show an increase of 1.66 mm (32%), this tendency was not statistically significant (P .05).Results suggest that volumetric heating of vaginal tissue produced quantitative improvement in the connective tissue organization in a porcine study. Neocollagenesis and neoelastogenesis were observed with an increased number of synthetically active cells.

Objectives While controlled thermal changes in subcutaneous tissue have been used to trigger apoptosis of fat cells and have been proven clinically efficacious, another mechanism of electromagnetic stress suggests that fat apoptosis could be achieved by a non-thermal manner as well. This animal model study investigates the use of a non-invasive high-intensity magnetic field device to induce apoptosis in fat cells. Methods Yorkshire pigs (N = 2) received one treatment (30 minutes) in the abdominal area using a High-Intensity Focused Electromagnetic (HIFEM) device. Punch biopsy samples of fat tissue and blood samples were collected at the baseline, 1 and 8 hours after the treatment. Biopsy samples were sectioned and evaluated for the levels of an apoptotic index (AI) by the TUNEL method. Statistical significance was examined using the rANOVA and Tukey's test (α 5%). Biopsy samples were also assessed for molecular biomarkers. Blood samples were evaluated to determine changes related to fat and muscle metabolism. Free fatty acids (FFA), triacylglycerol (TG), glycerol and glucose (Glu) were used as the main biomarkers of fat metabolism. Creatinine, creatinine kinase (CK), lactate dehydrogenase (LDH) and interleukin 6 (IL6) served as the main biomarkers to evaluate muscle metabolism. Results In treated pigs, a statistically significant increase in the apoptotic index (AI) (P = 1.17E-4) was observed. A significant difference was found between AI at baseline (AI = 18.75%) and 8-hours post-treatment (AI = 35.95%). Serum levels of fat and muscle metabolism indicated trends (FFA -0.32 mmol · l-1 , -28.1%; TG -0.24 mmol · l-1 , -51.8%; Glycerol -5.68 mg · l-1 , -54.8%; CK +67.58 μkat · l-1 , +227.8%; LDH +4.9 μkat · l-1 ,+35.4%) suggesting that both adipose and muscle tissue were affected by HIFEM treatment. No adverse events were noted to skin and surrounding tissue. Conclusions Application of a high-intensity electromagnetic field in a porcine model results in adipocyte apoptosis. The analysis of serum levels suggests that HIFEM treatment influences fat and muscle metabolism. Lasers Surg. Med. 51:47-53, 2019. © 2018 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

Clostridium difficile is a major nosocomial pathogen in humans with an increasing incidence in the community. The “one-health” approach of research is needed to investigate possible reservoirs of C. difficile and route of its transmission. The objective of this study is to investigate the occurrence of C. difficile in pigs in the Czech Republic with characterisation of the isolates to determine their genetic relatedness to C. difficile isolates from European and Asian pigs. A total of 198 pig faeces samples from 23 farms were investigated and of those 57 samples (55 piglets, 2 sows) from 11 farms were confirmed as C. difficile positive. The majority of C. difficile isolates belonged to the sequence type 11 and clade 5. The predominant ribotypes were 078 (n = 23), 078-variant (n = 5), 033 (n = 10) followed by RTs 150 (n = 7), 011 (n = 5), 045 (n = 4), 126, 014, 002 (n = 1, each). All isolates were susceptible to metronidazole, vancomycin and tetracycline. Isolates of RTs 150 and 078-variant were moxifloxacin resistant (MIC≥32 mg/L) and carried the amino acid substitution Thr82Ile in the GyrA. A multi-locus variable number tandem-repeats analysis (MLVA) revealed a clonal relatedness of isolates within individual farms and in C. difficile RT078 isolates between two Czech farms. Czech C. difficile RT078 isolates clustered with German C. difficile RT078 isolates and Czech C. difficile 078-variant isolates clustered with C. difficile RT078 isolates from Japan and Taiwan. This study found an emergence of C. difficile RT078 in Czech piglets that was related genetically to C. difficile RT078 isolates from Germany, Japan and Taiwan.

The aesthetic market offers various radiofrequency treatments for the reduction of wrinkles and rhytids. Even though this not an uncommon aesthetic therapy, there is considerable lack of clinical evidence on the various energy delivery systems available (unipolar, bipolar, tripolar, multipolar, etc.). The purpose of this study was to demonstrate the efficacy of a monopolar radiofrequency device (Exilis Elite, BTL Industries Inc., Boston, MA, USA) on the skin collagen in an animal model. The study treatment was done on the abdominal area of the potbellied Vietnamese mini pigs in the Veterinary Research Institute facility. All pigs were treated once per week for 4 weeks. The treatment area was sized 20 × 10 cm. The surface temperature was kept in the therapeutic interval from 39 °C to 43 °C and the therapy lasted for 10 minutes after reaching the therapeutic temperature. Biopsy samples were taken before the therapy and at the 3-month follow-up. The histology samples were stained and magnified (×400) before computer processing. The collagen volume was calculated using the stereological analysis and the data were statistically processed (using the nonparametric two-sample t-test). The collagen content tissue increased from average of 9.0% before the therapy up to 25.9% after the 3-month follow-up period. The statistical comparison of 54 samples taken before and after the treatment acknowledged the significant difference (p = 0.018). The stereological analysis proved large-scale improvement of collagen in the treated area. We have observed that the monopolar radiofrequency therapy significantly increases collagen remodeling.

The purpose of this study was to demonstrate the effect of monopolar, focused radiofrequency (RF) with embedded cooling on subcutaneous skin structures. Specifically, the study was to prove that the monopolar RF with cooling can selectively heat fat, causing disintegration of adipocytes and programmed cell death (apoptosis) of the subcutaneous fat cells.A non-invasive monopolar RF device with cooling (Exilis Elite, BTL Industries, Framingham, MA) was used to reduce abdominal fat in a porcine model. The study was done on 3 Vietnamese pigs in a certified veterinary facility. The treatment was delivered to an area the size of 20 x 10cm. The treatment duration was 11 minutes, 30 seconds. Biopsy samples were taken before the first treatment, 1 hour post each treatment, as well as 8 and 20 hours post each treatment. Programmed cell death (apoptosis) was monitored using the TUNEL method. The temperature was measured on the skin surface by an infrared thermal imager and built-in IR thermometer, and by an internal probe inserted into various depths of the subcutaneous layer. The internal probe placement was monitored by diagnostic ultrasound examination.The temperature in the treated adipose tissue was higher compared to the skin surface temperature. The average temperature gradient observed was 3.1°C. Due to the temperature gradient the skin surface remained intact, while subcutaneous layers showed significant changes. The TUNEL method proved large-scale apoptosis of fat cells after each treatment. The apoptotic index increased from 7% before the first treatment to an average of 53.4%, 39.6%, 40.2%, and 44.7% respectively for each treatment. In the three-month follow up the apoptotic index dropped back to 11.7%. Histology, blood biochemistry and hematology samples showed mild to no signs of inflammation in the treated area.The study has shown that use of monopolar, focused radiofrequency can induce substantial apoptotic process in a porcine model. The data suggests that the monopolar, focused radiofrequency device can be used for reduction of fat and body shaping.

Background: Selective fat reduction has been clearly shown for various methods and energy modalities including cryolipolysis and high intensity focused thermal ultrasound. Mathematical modeling of focused high frequency of the EM spectrum has indicated that selective heating of fat is possible using wavelengths not previous explored. The purpose of this study was to demonstrate in the porcine model that selective heating of fat is possible with a non-contact, operator independent device. Methods: High frequencies of the Industrial, Scientific and Medical (ISM) RF band were utilized to reduce abdominal fat in a porcine model. Practical application of mathematical modeling allowed an auto-feedback loop to be developed to allow operator independent adjustment of energy to maintain subcutaneous fat at 45–468C while overlying skin remained at 40–418C. Results: Treatments of three Vietnamese pigs were performed under anesthesia in a certified veterinary facility. Gross and microscopic histologic results demonstrated a marked reduction in adipocytes of the treated area after 4 treatments of a total of 30 minutes each, with incremental fat diminution after each treatment. A final 70% reduction of the abdominal fat layer was seen in the treated areas. Duplex ultrasound revealed a reduction of fat layer from 7.6 to 2.9 mm. Histologic evaluation revealed that epidermis, dermis, and adnexal structures such as hair follicles were unaffected by the treatment, while adipocytes were significantly affected. Conclusion: A new model of fat reduction using high frequency RF has been successfully achieved in a porcine model. This has very positive implications in the development of an operator independent, contact free device for reduction of fat in clinical practice. Lasers Surg. Med. 45:235–239, 2013. 2013 Wiley Periodicals, Inc.

The ability of different adjuvants to enhance immune responses to intradermal (ID) immunisation with a model antigen was studied in pigs. Immune responses were evaluated with respect to the intensity of systemic and mucosal antibody formation, their isotype characterisation and rate of cell-mediated immunity. These findings were compared with the intensity of adverse local reactions. Six groups of piglets were immunised with keyhole limpet haemocyanin (KLH) antigen alone or in combination with aluminium hydroxide or selected oil-based adjuvants (complete and incomplete Freund's adjuvants, Montanide ISA 206 and Emulsigen). Systemic specific antibody responses were significantly increased following ID administration of antigen together with any of the adjuvants used. IgG antibody responses were most pronounced after the first administration of antigen, being stimulated with both Freund's and Montanide ISA 206 adjuvants. The oil adjuvants also enhanced the cell-mediated immune responses and the levels of local IgA antibodies in the respiratory mucosa. On the other hand, they elicited more pronounced adverse local reactions.

The intensity of antibody responses and level of protection against challenge infection induced by Actinobacillus pleuropneumoniae (APP) were compared in piglets vaccinated intramuscularly with different vaccination doses of a subunit vaccine. Secondary antibody responses to APP exotoxins induced by the intradermally administered diluted vaccines did not differ from those induced by undiluted vaccines administered either intradermally or intramuscularly. The level of protection measured by the clinical course of challenge infection to the extent of lung lesions was significantly higher in animals vaccinated with a three-time diluted vaccine in comparison with animals that were administered intramuscularly or intradermally with the most concentrated or the most diluted vaccines.