Effect of p38 MAPK Inhibition on Apoptosis Marker Expression in the Process of Peritoneal Adhesion Formation

Background: Apoptosis, one of the most important mechanisms for maintaining homeostasis, is carried out under both physiological and pathological conditions. The aim of our study was to investigate the expression of markers of apoptosis through caspase-dependent and caspase-independent pathways during the reparative regeneration after serosal injuries of the peritoneum in the context of the prolonged p38 MAPK inhibition. Methods: Peritoneal adhesions in the animal models were induced by a method developed by the authors that included opening the serous-muscular layer of the caecum with a 1cm cut followed by closing the wound with a Schmieden suture and scarifying a 1.5×1.5 cm area of the parietal peritoneum of the right lateral channel. Experiments were carried out on male 9-month-old Wistar rats. When closing the wound, control animals were intraperitoneally administered 3 mL of 0.9% sodium chloride solution (n=40) while experimental group rats were administered slow-release drug Seroguard® (Pharmasyntez JSC) (n=40). Animals were sacrificed within the period of 2 hours to 30 days post surgery. Expression of apoptosis markers was studied by immunohistochemical (Bcl-2, Bcl-x) and immunofluorescent (PARP-1) staining. Results: It is interesting that, in cases of the natural regeneration of the peritoneal injury, expression of anti-apoptosis markers at the injury site came in two waves: it was the most pronounced on days 1–3 post surgery while the second peak of activity was observed on day 14. Within this time window, granulation tissue was actively growing and mature connective-tissue vascularized adhesions were being formed. By the end of the observation period (day 30), expression of anti-apoptosis proteins at the injury site became extremely low and a significant reduction in the amount of connective tissue cells was observed. It was found that a prolonged inhibition of the p38 activity resulted in a moderate increase in Bcl-2 expression on days 3–7, and a decrease in the activity on day 14 was followed by another increase in expression by day 30. The Bcl-xl expression was observed 12 hours to 3 days post surgery and then it went down to the minimum. Positive PARP-1 staining observed on days 3 to 30, which reached its maximum on day 14, was also typical of the experimental group. Conclusion: The performed study demonstrated that a prolonged p38 MAPK inhibition in the adhesion formation models results in the activation of fibroblast apoptosis at the reparation site, which, in the authors’ opinion, predetermines a significant decrease in the adhesion formation in the experimental group

Interleukin Expression in the Area damaged by the Development of Abdominal Cavity Adhesions

Background: This study sought to determine the dynamics of IL gene expression during serous membrane damage using an animal model of aseptic peritoneal injury. Methods: In our study, we used 35 male Wistar rats. Macroscopic and microscopic studies were conducted between 6 hours and 30 days after peritoneal damage was induced. In the damaged peritoneal area, we assessed IL gene expression across the experimental timeframe. Results: We found that the majority of the studied genes had three characteristic peaks in expression: at 6 hours, on day 3, and on day 14. These effects were observed for chemokine (CXC motif) ligands 1 and 3, IL1b, and IL6. Two peaks of increased expression (on days 3 and 14) were noted for CXCL1, CXCL5, INFγ, IL2, IL4, IL10, TNF, and CD40LG. Conclusion: We hypothesize that the absence of attention to the changes that occur in the peritoneum after aseptic damage has prevented research from focusing on the important stage of the formation of the richly vascularized adhesions that are unable to regress. Based on the results of our study, we conclude that it is critically important to influence the last wave of IL expression activation (2 weeks after aseptic peritoneum damage) to effectively prevent adhesion formation

Application of meso-CF3-Fluorophore BODIPY with Phenyl and Pyrazolyl Substituents for Lifetime Visualization of Lysosomes

A bright far-red emitting unsymmetrical meso-CF3-BODIPY fluorescent dye with phenyl and pyrazolyl substituents was synthesized by condensation of trifluoropyrrolylethanol with pyrazolyl-pyrrole, with subsequent oxidation and complexation of the formed dipyrromethane. This BODIPY dye exhibits optical absorption at λab ≈ 610–620 nm and emission at λem ≈ 640–650 nm. The BODIPY was studied on Ehrlich carcinoma cells as a lysosome-specific fluorescent dye that allows intravital staining of cell structures with subsequent real-time monitoring of changes occurring in the cells. It was also shown that the rate of uptake by cells, the rate of intracellular transport into lysosomes, and the rate of saturation of cells with the dye depend on its concentration in the culture medium. A concentration of 5 μM was chosen as the most suitable BODIPY concentration for fluorescent staining of living cell lysosomes, while a concentration of 100 μM was found to be toxic to Ehrlich carcinoma cells

Evaluation of the Safety and Toxicity of the Original Copper Nanocomposite Based on Poly-N-vinylimidazole

A new original copper nanocomposite based on poly-N-vinylimidazole was synthesized and characterized by a complex of modern physicochemical and biological methods. The low cytotoxicity of the copper nanocomposite in relation to the cultured hepatocyte cells was found. The possibility to involve the copper from the nanocomposite in the functioning of the copper-dependent enzyme systems was evaluated during the incubation of the hepatocyte culture with this nanocomposite introduced to the nutrient medium. The synthesized new water-soluble copper-containing nanocomposite is promising for biotechnological and biomedical research as a new non-toxic hydrophilic preparation that is allowed to regulate the work of key enzymes involved in energy metabolism and antioxidant protection as well as potentially serving as an additional source of copper

Efficiency of Direct Transcutaneous Electroneurostimulation of the Median Nerve in the Regression of Residual Neurological Symptoms after Carpal Tunnel Decompression Surgery

Carpal tunnel syndrome (CTS) is the most frequent entrapment neuropathy. CTS therapy includes wrist immobilization, kinesiotherapy, non-steroidal anti-inflammatory drugs, carpal tunnel steroid injection, acupuncture, and physical therapy. Carpal tunnel decompression surgery (CTDS) is recommended after failure of conservative therapy. In many cases, neurological disorders continue despite CTDS. The aim of this study was to investigate the efficiency of direct transcutaneous electroneurostimulation (TENS) of the median nerve in the regression of residual neurological symptoms after CTDS. Material and Methods: 60 patients aged 28–62 years with persisting sensory and motor disorders after CTDS were studied; 15 patients received sham stimulation with a duration 30 min.; 15 patients received high-frequency low-amplitude TENS (HF TENS) with a duration 30 min; 15 patients received low-frequency high-amplitude TENS (LF TENS) with a duration 30 min; and 15 patients received a co-administration of HF TENS (with a duration of15 min) and LF TENS (with a duration of 15 min). Results: Our research showed that TENS significantly decreased the pain syndrome, sensory disorders, and motor deficits in the patients after CTDS. Predominantly, negative and positive sensory symptoms and the pain syndrome improved after the HF TENS course. Motor deficits, reduction of fine motor skill performance, electromyography changes, and affective responses to chronic pain syndrome regressed significantly after the LF TENS course. Co-administration of HF TENS and LF TENS was significantly more effective than use of sham stimulation, HF TENS, or LF TENS in patients with residual neurological symptoms after CTDS

Efficiency of Direct Transcutaneous Electroneurostimulation of the Median Nerve in the Regression of Residual Neurological Symptoms after Carpal Tunnel Decompression Surgery

Carpal tunnel syndrome (CTS) is the most frequent entrapment neuropathy. CTS therapy includes wrist immobilization, kinesiotherapy, non-steroidal anti-inflammatory drugs, carpal tunnel steroid injection, acupuncture, and physical therapy. Carpal tunnel decompression surgery (CTDS) is recommended after failure of conservative therapy. In many cases, neurological disorders continue despite CTDS. The aim of this study was to investigate the efficiency of direct transcutaneous electroneurostimulation (TENS) of the median nerve in the regression of residual neurological symptoms after CTDS. Material and Methods: 60 patients aged 28–62 years with persisting sensory and motor disorders after CTDS were studied; 15 patients received sham stimulation with a duration 30 min.; 15 patients received high-frequency low-amplitude TENS (HF TENS) with a duration 30 min; 15 patients received low-frequency high-amplitude TENS (LF TENS) with a duration 30 min; and 15 patients received a co-administration of HF TENS (with a duration of15 min) and LF TENS (with a duration of 15 min). Results: Our research showed that TENS significantly decreased the pain syndrome, sensory disorders, and motor deficits in the patients after CTDS. Predominantly, negative and positive sensory symptoms and the pain syndrome improved after the HF TENS course. Motor deficits, reduction of fine motor skill performance, electromyography changes, and affective responses to chronic pain syndrome regressed significantly after the LF TENS course. Co-administration of HF TENS and LF TENS was significantly more effective than use of sham stimulation, HF TENS, or LF TENS in patients with residual neurological symptoms after CTDS

SARS-CoV-2 escape from cytotoxic T cells during long-term COVID-19

Here, the authors report accelerated intrahost evolution of SARS-CoV-2 in an immunocompromised patient with non-Hodgkin’s lymphoma with 318 days long COVID-19, and show that changes in the viral genome resulted in escape from T cellular immune response