Chronic Neurogenic Pain Research Articles

Electroneuromyographic examination of pelvic floor muscle dysfunction in patients with descending perineum syndrome

Aim: to assess the functional state of the pelvic floor muscles and their innervation in patients with descending perineum syndrome, manifested simultaneously by obstructive defecation and chronic neurogenic pelvic pain.Materials and methods: in 2022–2023, 51 patients (40 (78.4 %) women, 11 (21.6 %) men; age 50.9 ± 14.5 years) with pelvic organ prolapse, manifested by obstructive defecation and chronic neurogenic pelvic pain were examined at the Ryzhikh National Medical Research Centre for Coloproctology. All patients underwent high-resolution anorectal manometry and stimulation electroneuromyography according to a comprehensive neurophysiological protocol using a St. Mark’s electrode.Results: manometric patterns of functional defecation disorders were detected in 40 (78.4 %) patients, in the rest they were confirmed by defecography data and a negative evacuation test according to the Rome criteria IV. With stimulation electroneuromyography, neuropathy was detected in 68.6 % of cases by increasing the latency of the M-response of the pudendal nerve on the right or left. Complex neurophysiological diagnostics made it possible to additionally identify neuropathy in another 25.5 % of patients: according to changes in the mixed reciprocating reflex response (mixed feedback-reflex response) (efferent pathway) — in 17.7 % of cases; according to changes in the bulbocavernous reflex (efferent and afferent pathways) — in 7.8 % of cases. That is, neuropathy was determined in 94.1 % of patients in total.Conclusions:Functional diagnostic methods are an effective tool for studying the state of the pelvic floor muscle structures and their innervation and make it possible to identify pathogenetic links of disorders, the clinical manifestations of which are anal incontinence, neurogenic pelvic pain and proctogenic constipation in the form of obstructive defecation.Stimulation electroneuromyography makes it possible to detect disturbed innervation of the pelvic floor muscles and anal sphincter in the standard study of the M-response in 68.6 % of cases with perineal prolapse syndrome with a combination of proctogenic disorders and neurogenic pelvic pain. The use of a comprehensive neurophysiological protocol makes it possible to detect innervation disorders in 94.1 % of cases.

Mechanistic insights into the analgesic and anti-inflammatory effects of alcoholic extracts from Curcuma longa

Background: Aim of the study was to scientifically validate the traditional Indian claims of Curcuma longa's (turmeric) antinociceptive (pain-relieving) and anti-inflammatory effects. Methods: The alcoholic extract of C. longa was tested in three rodent nociceptive models: acetic acid-induced writhing: examines visceral pain, formalin test: evaluates both acute and chronic neurogenic and inflammatory pain and tail immersion test to assess thermal pain. The extract's effects were compared to a control group and morphine (reference drug). Results: C. longa extract significantly reduced abdominal constrictions in the acetic acid test (59.36% inhibition). In the formalin test, the extract significantly decreased paw licking response time in both early (54.12% inhibition) and late phases (78.59% inhibition). C. longa extract significantly increased the tail flick reaction time in the immersion test, indicating pain relief. Conclusions: This study confirms the antinociceptive and anti-inflammatory activities of C. longa, providing scientific evidence for its traditional use in pain management.

Complex neurophysiological algorithm for pudendal nerve neuropathy for descending perineum syndrome

Aim: to improve diagnostics for descending perineum syndrome.Patients and Methods: the prospective cohort study included 127 patients (85 (66.9%) — females), aged 49.9 ± 14.4 years with descending perineum syndrome. All patients had proctogenic constipation, anal incontinence and/or chronic neurogenic pelvic pain. All patients underwent a neurophysiological examination according to the original complex pudendal nerve terminal motor latency (PNTML) method — neurophysiological protocol for detection of the pelvic floor muscles innervation disorders.Results: the latency of the M-response of the pudendal nerve increased on at least one side in 85 (66.9%) patients (50 women and 35 men). The use of a new complex neurophysiological diagnostic protocol made it possible to identify signs of pudendal neuropathy in 29.9% of patients. The incidence of neuropathy in patients with clinically significant perineal prolapse syndrome was 96.5% in females and 97.6% in males.Conclusion: the new complex neurophysiological diagnostic algorithm made it possible to identify disorders of innervation along the efferent pathway in the Alcock’s canal and distally in 85 (66.9%) cases, along the efferent pathway proximal to the Alcock’s canal — in 23 (18.1%); a combination of efferent and afferent disturbances occurred in 15 (11.8%).

MORPHOLOGICAL ASPECTS OF DEVELOPMENT OF MALIGNANT TUMORS UNDER THE CONDITIONS OF COMORBID PATHOLOGY AND MITOCHONDRIAL THERAPY

Morphological studies represent the most important part of evidence-based medicine, and in oncology they are an essential attribute in the diagnostics of malignant neoplasms. In experimental oncology, current models of the tumor growth are developed to be as close as possible to the actual biological life conditions. There is a need to study the pathogenesis of tumors in various variants of their orthotopic growth, the formation of bi-model systems with a combination of the malignant growth and comorbid conditions (chronic neurogenic pain, diabetes mellitus, hypothyroidism, obesity), the use of subcellular substrates for the induction of carcinogenesis and biotherapy. Among the important methods for studying the pathogenesis of tumors, morphology is on a par with advanced molecular genetic and biochemical methods, as well as radioimmunoassay techniques.Morphological studies represent the most important part of evidence-based medicine, and in oncology they are an essential attribute in the diagnostics of malignant neoplasms. In experimental oncology, current models of the tumor growth are developed to be as close as possible to the actual biological life conditions. There is a need to study the pathogenesis of tumors in various variants of their orthotopic growth, the formation of bi-model systems with a combination of the malignant growth and comorbid conditions (chronic neurogenic pain, diabetes mellitus, hypothyroidism, obesity), the use of subcellular substrates for the induction of carcinogenesis and biotherapy. Among the important methods for studying the pathogenesis of tumors, morphology is on a par with advanced molecular genetic and biochemical methods, as well as radioimmunoassay techniques.

ID: 203889 Inactivation of Spinal Cord Stimulator Implanted Pulse Generators After Elective Surgery: An Under-Recognized Problem

Spinal cord stimulation (SCS) is an established and effective method for chronic neurogenic pain relief consisting of the placement of an epidural lead and implanted pulse generator (IPG). The electronics of SCS IPGs are susceptible to damage when implanted patients undergo surgery. Some SCSs have a dedicated surgery mode, while others recommend turning the SCS off to protect it from damage. IPG inactivation may require resetting or replacement surgery and the prevalence of this real-world problem has not been studied.

Analysis of changes in the serotonin inactivation coefficient in brain structures with simultaneous modeling of chronic neurogenic pain and malignant neoplasia

Introduction. Chronic pain syndrome in the development of malignant neoplasms has a complex polyethological character. The study of the pathogenetic mechanisms of pain during the growth of the transplanted tumor in the experiment can open up broad perspectives for the creation of new domestic analgesic drugs for use in oncology.Aim: To study the serotonin inactivation coefficient (SIC), the ratio of serotonin (S) and its main metabolite 5-hydroxyindolacetic acid (S-OH) in brain areas (cortex, hypothalamus) with simultaneous modeling of chronic neurogenic pain and malignant neoplasia.Material and Methods. The study involved white mongrel rats (males). To create a model of chronic pain (CP), ligation of the sciatic nerve was performed according to the method of V.V. Kravtsova in modification. Rat sarcoma M-1 was used to simulate malignant neoplasia. At the main stage of the study, the animals were divided into 4 groups: control, two comparison groups (model of chronic pain, standard M-1 sarcoma grafting) and experimental (combined modeling of CP and oncological process). Animals of the second, third and fourth groups were euthanized sequentially at the 2nd and 3rd weeks of the development of the tumor process.Results. Chronic pain syndrome disrupts the process of serotonin metabolism: the decrease in SIC in the cerebral cortex of experimental animals was more significant by 48% (day 14) and 72% (day 21). The development of the oncological process in comparison groups (groups 3a and 3b) is accompanied by a decrease in the level of SIC, more significant at the 21th day of M-1 sarcoma development: by 37% in the hypothalamus, by 41% in the cerebral cortex. With simultaneous modeling of chronic neurogenic pain and malignant neoplasia, the lowest values of the studied indicator were noted, the decrease was 75% in the cerebral cortex (group 4a) and 87% (group 4b).Discussion. Chronic neurogenic pain and M-1 sarcoma development in isolated modeling variants cause a significant decrease in SIC in hypothalamus and cerebral cortex due to disorders of serotonin formation. A more significant disruption of the serotonin mediator system was noted in cerebral cortex which leads to a decrease in the body’s adaptive capabilities to pain and a disruption of the regulatory mechanisms of metabolism.Conclusion. Modification of serotonin metabolism can be considered as a potential therapeutic target for the treatment of chronic pain syndrome in oncology.

Analysis of changes in the serotonin inactivation coefficient in brain structures with simultaneous modeling of chronic neurogenic pain and malignant neoplasia

Introduction. Chronic pain syndrome in the development of malignant neoplasms has a complex polyethological character. The study of the pathogenetic mechanisms of pain during the growth of the transplanted tumor in the experiment can open up broad perspectives for the creation of new domestic analgesic drugs for use in oncology.Aim. To study the serotonin inactivation coefficient (SIC), the ratio of serotonin (S) and its main metabolite 5-hydroxyindolacetic acid (S-OH) in brain areas (cortex, hypothalamus) with simultaneous modeling of chronic neurogenic pain and malignant neoplasia.Material and Methods. The study involved white mongrel rats (males). To create a model of chronic pain (CP), ligation of the sciatic nerve was performed according to the method of V.V. Kravtsova in modification. Rat sarcoma M-1 was used to simulate malignant neoplasia. At the main stage of the study, the animals were divided into 4 groups: control, two comparison groups (model of chronic pain, standard M-1 sarcoma grafting) and experimental (combined modeling of CP and oncological process). Animals of the second, third and fourth groups were euthanized sequentially at the 2nd and 3rd weeks of the development of the tumor process.Results. Chronic pain syndrome disrupts the process of serotonin metabolism: the decrease in SIC in the cerebral cortex of experimental animals was more significant by 48% (day 14) and 72% (day 21). The development of the oncological process in comparison groups (groups 3a and 3b) is accompanied by a decrease in the level of SIC, more significant at the 21th day of M-1 sarcoma development: by 37% in the hypothalamus, by 41% in the cerebral cortex. With simultaneous modeling of chronic neurogenic pain and malignant neoplasia, the lowest values of the studied indicator were noted, the decrease was 75% in the cerebral cortex (group 4a) and 87% (group 4b).Discussion. Chronic neurogenic pain and M-1 sarcoma development in isolated modeling variants cause a significant decrease in SIC in hypothalamus and cerebral cortex due to disorders of serotonin formation. A more significant disruption of the serotonin mediator system was noted in cerebral cortex which leads to a decrease in the body's adaptive capabilities to pain and a disruption of the regulatory mechanisms of metabolism.Conclusion. Modification of serotonin metabolism can be considered as a potential therapeutic target for the treatment of chronic pain syndrome in oncology.

ПОЛОВЫЕ РАЗЛИЧИЯ В СОДЕРЖАНИИ СТЕРОИДНЫХ ГОРМОНОВ В МИТОХОНДРИЯХ КЛЕТОК СЕРДЦА НА ЭТАПАХ РАЗВИТИЯ МЕЛАНОМЫ В16/F10, СОПРЯЖЕННОЙ С ХРОНИЧЕСКОЙ НЕЙРОГЕННОЙ БОЛЬЮ

Aim - an analysis of levels of steroid hormones in heart cell mitochondria at the stages of B16/F10 melanoma development in combination with chronic neurogenic pain in male and female C57BL/6 mice. The study included male and female C57BL/6 mice (n=336). Experimental groups were: intact animals (♂ n=21; ♀ n=21); controls (♂ n=21; ♀ n=21) with a model of chronic neurogenic pain (CNP); the comparison group (♂ n=63; ♀ n=63) with melanomas (B16/ F10); the main group (♂ n=63; ♀ n=63) (CNP+B16/F10) with melanoma transplanted 3 weeks after the CNP model creation. Levels of estradiol (pg/g protein), estrone (pg/g protein), progesterone (ng/g protein), total testosterone (ng/g protein), and free testosterone (pg/g protein) were determined by ELISA in mitochondrial samples. In animals with CNP, changes in levels of steroid hormones were detected only in heart mitochondria of female mice: estradiol was reduced by 4.6 times, estrone by 2.2 times, progesterone by 1.8 times (p˂0.05), and total testosterone by 2.4 times compared with intact values. Melanoma growth (3 weeks) in females downregulated levels of estradiol by 1.7 times (p˂0.05), estrone by 1.8 times (p˂0.05), progesterone by 2.1 times, and total testosterone by 2.3 times compared with intact values. In males, only total testosterone decreased by 2.1 times during the same period. Melanoma growth in presence of CNP (3 weeks) in females decreased levels of estradiol by 1.6 times (p˂0.05) compared with control val- ues, while males showed reduced levels of estradiol and total and free testosterone by 1.3 times (p˂0.05), 1.9 times (p˂0.05) and 2.7 times, respectively, as well as elevated levels of estrone by 1.6 times (p˂0.05). CNP and the growth of B16/F10 melanoma to a greater extent weaken the heart of females due to the involvement of a wider spectrum of hormones and a decrease in their levels in the heart mitochondria, compared to males. When CNP was combined with B16/F10 melanoma, a more significant hormonal imbalance was determined in males than in females.

Assessment of Amnesic Effect and the Depth of Hypnosis During Therapeutic Inhalation of Xenon-Oxygen Mixture

The current literature lacks data on the incidence of intraoperative awakening during xenon anesthesia. This could be due to amnesia preventing memories of the intraoperative awakening.The objective: to determine the concentration of xenon in the xenon-oxygen mixture, which causes amnesia for events during inhalation in 100% of patients, and to make correlations with the depth of hypnosis as per Kugler scale.Subjects and Methods: 34 patients with chronic neurogenic pain who received 111 20-minute inhalations with concentration of xenon up to 50% were included in the study. Amnesia evaluation, EEG monitoring, and pain assessment on a visual analog scale (VAS) were performed.Results. Amnesic effect developed in 100% of patients at xenon concentration of 45%. On inhalation of xenon at concentrations of up to 50%, EEG changes did not exceed D1 grade on Kugler scale. The decrease in bispectral index (BIS) did not reach the level of deep sedation (Me 96.2%) at xenon concentration of 50%. The decrease in pain on VAS was approximately 60%.Conclusions. Xenon inhalations cause transient congradic amnesia at concentrations of 45% or more. The accuracy of the BIS monitoring readings may be reduced when using xenon in a monovariant. Inhalations of xenon-oxygen mixture in concentrations up to 50% showed good analgesic properties in the framework of combined therapy of chronic pain syndrome.

Chronic neurogenic pain inducing chemiluminescence of mitochondrial associates in cardiomyocytes.

e21560 Background: Mitochondrial dysfunction is has a significant impact on the development of heart disease and is a characteristic feature of the heart physiopathology. Our purpose was to analyze some mechanisms of apoptosis regulation and self-organization in the mitochondria of the heart cells in mice with melanoma growing in presence of chronic neurogenic pain (CNP). Methods: The study included female C57BL/6 mice (n = 105) divided into groups: intact animals (n = 21); controls (C, n = 21) with a CNP model created by bilateral sciatic nerve ligation under xylazine/zoletil anesthesia; main group (CNP+B16/F10, n = 63) with B16/F10 melanoma transplanted after CNP creation. After decapitation, mitochondria of the heart were isolated by differential centrifugation. Levels of cytochrome C (ng/mg of protein), caspase 9 (ng/mg of protein), Bcl-2 (ng/mg of protein), AIF (ng/mg of protein), Ca2+ (mmol/g of protein) were determined in mitochondrial samples by ELISA. Results: CNP downregulated levels of Ca2+ by 3.2 times, Bcl-2 by 1.3 times (p < 0.05) and caspase 9 by 1.5 times (p < 0.05), compared to intact mice. AIF levels, on the contrary, were elevated by 2.3 times, and cytochrome C did not differed statistically significantly from intact values. After a week of B16/F10 growth in presence of CNP, levels of Ca2+ in the mitochondria of the heart increased by 5.3 times relative to C values. Further on, Ca2+ decreased to almost undetectable values. The AIF levels changed abruptly: after 1 week it increased by 3.7 times, after 2 weeks it declined to C levels, and after 3 weeks it became 5.2 times lower than in C and 2.3 times lower than in intact animals. Bcl-2 and cytochrome C changed similarly: Bcl-2 after 1 week of melanoma growth in presence of CNP increased 1.7 times (p < 0.05) compared to C, and then after 2-3 weeks it declined and became on average 2.2 times lower than in the mitochondria of the C group; levels of cytochrome C after 1 week did not differed significantly from the values in C, and after 2-3 weeks they decreased by 2.2 times. The levels of caspase 9 in CNP+B16/F10 were on average 2.4 times higher than C values throughout the study. The cold light of high brightness - chemiluminescence was recorded in samples of mitochondrial suspension after 2-3 weeks of CNP+B16/F10. The glow in heart mitochondrial samples was accompanied by bright flashes and a 10-15 second intense white glow with a gradual fading and settling of a large filamentous aggregation of mitochondria on the substrate layer. CNP contributed to the energy supply system blocking in the energy systems of cardiomyocytes. Conclusions: Mitochondrial mechanisms of apoptosis and self-organization of subcellular energy structures in the conditions of malignant tumor growth in presence of CNP are mediated by disruption of polyenzymatic apoptosis regulation systems, and a high level of oxidative stress that induces chemiluminescence of cardiomyocyte mitochondrial associates.

Whose mitochondria are more stable, male or female ones? Response of neurosteroid status of mitochondria of cerebral cortex cells to melanoma development in presense of chronic pain.

e21559 Background: Gender differences in brain physiology and gender differences in pathology are usually recognized, but often are disregarded in clinical and experimental studies, resulting in numerous inaccuracies in data interpretation. The purpose of this study was to analyze levels of neurosteroid hormones in mitochondria of the cerebral cortex cells in C57BL/6 mice with subcutaneous B16/F10 melanoma growing in presence of chronic neurogenic pain (CNP). Methods: The study included male and female C57BL/6 mice (n = 336) aged 8 weeks initially weighing 21-22 g. Experimental groups were: intact animals; controls with a CNP model created by bilateral sciatic nerve ligation; comparison group 3 weeks after subcutaneous inoculation of 0.5 ml suspension of B16/F10 melanoma cells diluted 1:10; main group 3 weeks after subcutaneous melanoma growth in presence of CNP (CNP+B16/F10). Levels of estradiol (pg/g protein), estrone (pg/g protein) (DBC, Canada); progesterone (ng/g protein), total and free testosterone (pg/g protein) (XEMA, Russia) were measured in mitochondrial samples by ELISA (Tecan Infinite F50 analyzer, Austria). Results: Levels of estradiol in intact females were 3.1 times higher than in males, while estrone, progesterone, total and free testosterone were lower by 6.4, 2.7, 2.0 and 2.5 times, respectively. Only in females with CNP estradiol decreased by 3.4 times, compared to intact values, and estrone increased by 1.7 times (p<0.05), testosterone by 3.9 times. Estradiol in animals with B16/F10 decreased compared with intact values by 3.1 times in females and by 1.5 times in males (p<0.05). Females with B16/F10 showed the highest levels of progesterone exceeding intact values by 3.1 times. In the group with CNP+B16/F10, females showed lower levels of estradiol and estrone (by 1.4 times, p<0.05) and free testosterone (by 1.6 times, p<0.05), compared to the levels in CNP; on the opposite, males had 1.5 times (p<0.05) higher estrone and 2.9 times lower progesterone. Conclusions: Low levels of estradiol involved in the protective mechanism of neurosteroids were the dominant factor in mitochondria of cerebral cortex cells in females with CNP and malignant neoplasms. Males did not demonstrate such dominant factor. In animals with CNP+B16/F10, the response nature of cerebral cortex mitochondria changed: all defense mechanisms in the brain of females were suppressed by the 3rd week of tumor development in presence of pain. In males, mitochondria of cerebral cortex cells were more resistant to the influence of two pathologies, and only few changes in the neurosteroid status were recorded.

Attacking effect of diabetes mellitus and chronic neurogenic pain on malignant process:Are mitochondria of the heart damaged?

e21557 Background: Mitochondrial dysfunction is one of the mechanisms for the development of heart disease that requires close study. Diabetic cardiomyopathy is the leading cause of death in such patients due to the development of heart failure. Chronic pain may be associated with higher prevalence of cardiovascular disease, and little is known about its potential biological consequences. The purpose of this study was to analyze parameters of free radical oxidation and mitochondrial respiration in heart cells in experimental animals with malignant tumors growing in presence of diabetes mellitus (DM) and chronic neurogenic pain. Methods: The study included outbred female rats (n = 32) weighing 180-220 g and C57BL/6 female mice (n = 84) weighing 21-22 g. Experimental groups of rats were: intact animals (n = 8), controls (n = 8) with DM, comparison group (n = 8) with subcutaneously inoculated Guerin’s carcinoma, and main group (n = 8) with Guerin’s carcinoma subcutaneously inoculated after 1 week of persistent hypothyroidism. Experimental groups of mice were: intact animals (n = 21), controls (n = 21) with a model of chronic neurogenic pain (CNP) created by bilateral sciatic nerve ligation, comparison group (n = 21) with subcutaneously inoculated melanoma (B16/F10), and main group (n = 21) with melanoma subcutaneously inoculated 3 weeks after the CNP model was created (CNP+B16/F10). Heart mitochondria were isolated by differential centrifugation. Levels of cytochrome C (ng/g of protein), 8-hydroxy-2'-deoxyguanosine (8-OHdG) (ng/g of protein), and malondialdehyde (MDA) (nM/g of protein) were measured in mitochondrial samples by ELISA. Statistical analysis was performed using the Statistica 10.0 program. Results: DM in rats upregulated 8-OHdG by 6.3 times and MDA by 1.9 times (p < 0.05) and downregulated cytochrome C by 1.5 times (p < 0.05) in heart cell mitochondria, compared to intact values. DM+Guerin’s carcinoma in rats increased 8-OHdG by 14.0 times and MDA by 1.7 times (p < 0.05) and decreased cytochrome C by 1.5 times (p < 0.05), compared to intact values. CNP in mice did not affect the studied parameters in mitochondria of the heart. CNP+B16/F10 in mice increased 8-OHdG by 7.1 times and MDA by 1.6 times (p < 0.05) and decreased cytochrome C by 1.6 times (p < 0.05) in heart cell mitochondria. Conclusions: Comorbidity (diabetes mellitus, chronic neurogenic pain) together with malignant pathology aggravates mitochondrial dysfunction of heart cells which results in DNA damage and destabilization of the respiratory chain mediated by free radical oxidation processes.

Antitumor pharmacological activity of hybrid organotin compounds on the Lewis epidermoid carcinoma model.

e15004 Background: New antitumor agents based on organic metal derivatives are intensively determined and created by now. Tin compounds are of great interest among them [Milaeva E.R.et al. Novel selective anticancer agents based on Sn and Au complexes. Pure & Applied Chemistry. 2020, 92, 1201-1216]. The results of studies on cell line models allowed us to conclude that cytotoxic effect of organotin compounds is pronounced, so it is advisable to hold their further studies on biological models in vivo [ Dodokhova M.A. et al. Pharmacotherapeutic potential’s evaluation of organotin compounds in vivo Biopharmaceutical journal, 13(3), 30 – 34 (2021); doi: 10.30906/2073-8099-2021-13-3-30-34]. For the specialists of experimental pharmacology and oncology is relevant to rate the antitumor activity of hybrid organotin compounds on experimental malignant tumors. Methods: The study was held on C57Bl/6 mice (females, n = 36, each cohort contained 12 mice) on a Lewis epidermoid carcinoma model (subcutaneous transplantation) [Kit O.I. et al. Influence of chronic neurogenic pain on the dynamics of the no-system functioning during melanoma B16/F10 growth in male mice. Siberian Journal of Oncology, 2021, 20(3), 67–75]. Bis-(3,5-di-tert-butyl-4-hydroxyphenyl)dimethyl tin(Me-3) and (3,5–di–tert–butyl–4–hydroxyphenylthiolate) triphenyl tin (Me-5) were used as chemotherapeutic agents. Groups of animals were formed as follows: I (experimental) - Me-3 at a dose of 75 mg / kg; II (experimental) - Me-5 at a dose of 50 mg / kg; III (control group) - without pharmacological substances. The doses were characterized as the most effective according to the results of a pharmacological activity screening study. The compounds were injected intraperitoneally daily for 5 days. All animals were euthanized since 21 days after the tumor cells transplantation. The antitumor effect was estimated by the percentage of tumor growth inhibition by weight (%). All manipulations were carried out in accordance with the European Convention for the Protection of Vertebrates Used for Experimental and Other Scientific Purposes (ETS 123). Results: After the intraperitoneal injection of chemical substances Me-3 and Me-5, there was revealed an inhibition of growth by weight of Lewis epidermoid carcinoma’s primary node for all experimental groups, 23.7% and 28% respectively. Conclusions: The injection of organotin compounds with different ligand groups allows us to suggest a biocidal activity of the organotin component on the primary tumor node cells.

CAMP сoncentrations in cardiac mitochondria and serum in the С57ВL/6 mice under independent melanoma В16/F10 growth versus melanoma В16/F10 growth linked to chronic neurogenic pain

The aim of this research work is to study the cAMP level in the cardiac mitochondria and serum in the С57ВL/6 strain mice of both genders under the independent melanoma В16/F10 growth versus the melanoma В16/F10 growth linked to chronic neurogenic pain (CNP). Materials and methods. Mice of strain С57ВL/6 (n=336) have been grouped as follows: the intact group of the mice (♂n=21; ♀n=21), the reference group (♂n=21; ♀n=21) with the reproduced CNP model, the comparison group (♂n=63; ♀n=63) to include the mice with melanoma В16/F10, and the main test group (♂n=63; ♀n=63) to cover the mice with the melanoma growth against the CNP background. Upon expiration of 1 week, 2 and 3 weeks of the melanoma growth, in the animals of the above experimental groups the cardiac mitochondria have been isolated with the centrifugation using high-performance refrigerated centrifuge Avanti J-E, BECMAN COULTER, USA. With ELISA Kit (RayBio USA) we have determined cAMP concentrations in serum and in the cardiac mitochondria. Results. CNP has induced a decrease in the cAMP level in the cardiac mitochondria by a factor of 3,6 in the female mice only. In the animals of the comparison group the cAMP level in the heart has been increasing beginning with week 2 of the tumor growth on average by a factor of 4, while in the main test group starting from week 1 of the tumor growth it has been recorded 2-4 times higher and was depleted by the end of the experiment. As to the cAMP concentration in serum, the dynamics thereof has not been found to be in correlation with the cardiac mitochondrial data, and its concentration decrease has been recorded both in the females and the males. Conclusion. So, the changes in the cAMP concentration in the cardiac mitochondria demonstrate their gender-specific feature; the female mice as against the males have responded to an independent impact produced by CNP. As to the main test group, CNP has stimulated an increase in the cAMP level in the cardiac mitochondria 1 week earlier than it is the case with the comparison group, and it has resulted in the full cAMP depletion by the 3rd week of the experiment.

Structural myocardial catastrophe under the influence of chronic neurogenic pain due to development of B16 melanoma in female mice

Topicality. Most of the inhabitants on the planet have directly experienced certain manifestations of pain, and in the case of chronic pain sensations, some significant changes may occur not only in the psycho-emotional sphere of a human individual, but also some functional alterations in the life maintenance systems. There are two global problems of medicine which are associated with oncology and cardiovascular pathology, and, taking into account the prevalence of pain symptoms of various etiologies, a pathological triangle is formed, each side of which affects the overall configuration thereof. There is a need to understand these pathological relationships in order to determine the real possibilities of breaking them and correcting such common complex life situations. Any attempt to create a concept and visually verify the real pathological changes in the heart, when modeling a combination of chronic neurogenic pain and a malignant process, is relevant that is the aim of this research work. We targeted the morphological picture of the heart in female mice with chronic neurogenic pain (CNP) and the growth of B16 melanoma and found a rather tough scenario of unfolded events. Materials and methods. We used female mice of the C57BL/6 line with a normal genotype. Chronic neurogenic pain (CNP) was produced by bilateral ligation of the sciatic nerve. Against that background, all animals were subcutaneously transplanted with melanoma B16/F10. After decapitation, the isolated heart preparations were carried out according to the stages of morphological preparation for staining sections with hematoxylin-eosin, followed by morphological examination of the structure with the Leica DM LS2 microscope furnished with an Olympus optical.C-5050 Zoom video camera and Morfotest software. Photographing was conducted with magnifications x10, x40, x100. Results. A pronounced pro-oncogenic effect of pain stimulation of the neurogenic nature was revealed, which consisted in an earlier manifestation of the tumor growth, large-scale metastasizing even to atypical target organs, and the formation of a pre-terminal state at an earlier time. At the same time, morphological correlates of prolonged damage to the heart at the level of the valves and the ventricular wall were determined, the key elements of which were ischemia, total longitudinal splitting of muscle fiber bundles, blood filling of large vessels, hemorrhages, deep cell dystrophy of cardiomyocytes, myolysis, macrofocal necrosis (myomalacia), an accumulation of necrotic masses, fibrosis and eosinophilic infiltration. Visualization of the nuclei of cardiomyocytes indirectly indicated the switch of the death program to the non-apoptotic pathway, i.e. necrosis as a result from the “tumor-caused” depletion of the energy reserves of cardiomyocytes. Conclusion. The simulated hypertrophied manifestations of myocardial catastrophe in the experiment expand the concept of a prolonged infarction and provide a basis for predicting and preventing a negative course of events in difficult patients with persistent pain syndromes and comorbid pathology against the background of the development of a malignant process.

Biological effects of mitochondrial therapy: preventing development of myocardial infarction and blocking metastatic aggression of B16/F10 melanoma

The aim is to evaluate the physiological parameters of the efficacy of cardiac mitochondria transplantation in male mice with chronic neurogenic pain and B16/F10 melanoma growth. Materials and methods. Male mice (n=37) of the C57BL/6 line were used in the research work. The animals covered experimental groups as follows: mice with chronic neurogenic pain (CNP) + B16/F10 melanoma (n=27); mice with CNB + B16/ F10 melanoma + mitochondrial therapy (MC therapy) (n=10). Mitochondria were isolated from the heart of an intact rat with the use of differential centrifugation. An introduction of mitochondria to mice was carried out daily intraperitoneally at a dose of 3.3 mg of protein for 3 weeks. Statistical analysis of the results is carried out with the Statistica 10.0 software. Results. On day 21 (week 3) of the experiment, macroscopically in the melanoma tissue in the group of animals with MC therapy, there were 2.5 times more necrosis cases than in the group without MC therapy. During the examination of the internal organs, no metastases were detected in the animals treated with MC therapy, while in 100% of the animals without MC therapy metastases were found in the lungs and in 95% of them in the spleen. In the animals received MC therapy, there was no damage to the heart muscle in 75% of the cases, while in the group of the animals without MC therapy, the presence of lesions in the form of bruises on the surface of the heart was macroscopically detected in 100% of the animals. Conclusion. Thus, intraperitoneal transplantation of intact heart mitochondria contributed to the prevention of myocardial infarction and metastases to internal organs in the C57BL/6 male mice with B16/F10 melanoma growing against the background of chronic neurogenic pain.

Mitochondrial therapy: direct visual assessment of the possibility of preventing myocardial infarction under chronic neurogenic pain and B16 melanoma growth in the experiment

On models of chronic neurogenic pain (CNP) and the growth of a malignant tumor (metastasizing B16 melanoma) in male mice, we studied an effect produced by mitochondrial therapy (MCT) on the state of the myocardium. Some structural correlates of the compensatory-restorative effect by mitochondria transplanted from healthy recipient rats were revealed. It has been identified that MCT contributes to the preservation of the structural integrity of the myocardial tissue, the inclusion of an auxiliary link in the cellular mechanisms of tissue restoration: fibroblasts, histiocytes, lymphocytes, eosinophils and other connective tissue elements, which implement the intercellular mechanism of information transfer that provides the external regulatory function of MCT. The ability of mitochondria to prevent the DNA decay determines the possibility of initiation of the operation of the nuclear mechanisms of the cardiomyocyte division, which is characteristic of a population of young cells and which indicates the determining position of exogenous mitochondria.

Levels of apoptosis factors in mitochondria of brain cortex cells in female С57ВL/6 mice in dynamics of B16/F10 melanoma growth combined with comorbidity

Purpose of the study. To analyze the apoptosis indicators in mitochondria of brain cortex cells in female С57ВL/6 mice in the dynamics of B16/F10 melanoma growth alone and in combination with comorbidity, i.e. chronic neurogenic pain.Materials and methods. Female С57ВL/6 mice (n = 168) were used in the experiment. Groups accounted: intact group (n = 21); control group (n = 21) with a model of chronic neurogenic pain (CNP); comparison group (n = 63) with B16/F10 melanoma transplanted subcutaneously; main group (CNP + B16/F10) (n = 63). Levels of cytochrome C (ng/mg protein), caspase 9 (ng/mg protein), Bcl‑2 (ng/mg protein), AIF (ng/mg protein), calcium (Ca 2+) (mMol/g protein) were measured by ELISA in mitochondrial samples. Statistical analysis was performed using the Statistica 10.0 program.Results. In a week of the tumor growth in presence of comorbidity, i.e. CNP, levels of calcium in murine brain cortex mitochondria were 1.4 times higher (p < 0.05) than in the comparison group at the same time; in 2 weeks the levels declined by 80.1 times and after 3 weeks declined by 37.7 times. Compared to the values in the comparison group AIF levels in animals with CNP+B16/F10 were lower by 25 and 1.8 times (p < 0.05) at weeks 1 and 3, respectively. Higher levels of Вcl‑2 in the group with CNP + B16/F10 were registered at weeks 2 and 3 by 2 and 1.4 times (p < 0.05), respectively. Levels of cytochrome C were decreased in animals with CNP+B16/F10 at weeks 1–3 by 3.2, 1.5 (p < 0.05) and 2.8 times, respectively. Caspase 9 in CNP+B16/F10 after 3 weeks exceeded the values in the comparison group by 2.6 times.Conclusions. Combination of CNP and melanoma at an early stage in the animal body promotes the accumulation of calcium and suppression of AIF and cytochrome C in mitochondria of the brain cortex. By the terminal stage of tumor growth in presence of comorbidity (CNP), suppression of most units of the respiratory chain of mitochondria of brain cortex cells is formed.

Mechanisms of cancellation by pain of genetically determined inhibition of a malignant tumor growth in experiment

Aims are to study the nature of the processes of carcinogenesis of experimental B16/F10 melanoma in uPA gene knockout mice modified by chronic neurogenic pain and investigate some electrophysiological mechanisms of melanoma development. Materials and methods. We used 48 C57BL/6-PlautmI. IBug-ThisPlau6FDhu/GFDhu mice of both genders with urokinase gene knockout and 102 C57BL/6 mice of both genders with the normal genotype. Chronic neurogenic pain (CNP) was produced due to bilateral ligation of the sciatic nerve. Against the above background, all animals were transplanted with B16/F10 melanoma. To study the mechanism of CNP, studies of the intracellular electrophysiological activity of neurons of the central nervous system of the snail Helix pomatia in the body in vivo were carried out. CNP was reproduced by dosed pressing of four main nerves with Fresnel hairs that with time turned into increasing pain. Membrane potential (MP), action potential (AP) and firing rate (FR) parameters of intracellular bio-potentials of the command neuron RPaG3, continuously recorded using an ultrathin glass microelectrode for 4-5 days, were analyzed. Results. It was detected that an activation of cancerogenesis during the modification of the progression of experimental B16/ F10 melanoma in C57BL/6-PlautmI.IBug-ThisPlau6FDhu/GFDhu mice with uPA gene knockout using CNP is accompanied by a 2-fold acceleration in the time of tumor production, stimulation of the growth of the primary tumor nodes from 1.05±0.08 cm3 to 9.50±0.98 cm3 (p<0.001) and multiple metastasizing to the lungs, a reduction in the life span from 34.67±0.67 to 21.33±2.19 days (p<0.001) <0.05) in the genetically modified mice, by changing some gender-specific characteristics of the progression of the malignant process. The neuropathic nature of pain resulting from command neuron compression or ligation of the sciatic nerves is essentially identical to the implementation of genetic programs responsible for the control of life and death, reproducing events in carcinogenesis with the progression of a malignant tumor. Conclusion. The initiation and chronization of pain at the local level of the nervous system can lead to generalization of the pain syndrome and contribute to the cancellation of genetically predetermined programs of carcinogenesis.

Changes in levels of urokinase receptor and other components of fibrinolytic system in brain tissues in urokinase gene-knockout mice with B16/F10 melanoma growing together with chronic neurogenic pain

Purpose of the study. An analysis of the changes in components of the urokinase system in the brain of urokinase gene-knockout mice (uPA-/-) with B16/F10 melanoma growing alone and together with chronic neurogenic pain (CNP).Materials and methods. The study included male and female C57BL/6-PlautmI.IBug-ThisPlau6FDhu/GFDhu mice (uPA-/-) (n = 48) and C57BL/6 mice (uPA+/+) (n = 80) with transplanted B16/F10 melanoma growing solitarily and together with CNP. Levels of the urokinase receptor (uPAR) and plasmin (PAP) and activity and levels of the PAI-I inhibitor were measured in the brain of animals by ELISA.Results. Levels of uPAR, PAI-I and PAP in the brain differed only in intact uPA-/- males, being on average 1.6 times higher (p < 0.05) than in uPA+/+ mice. Among animals with CNP, uPA-/- males showed increased PAI-I by 1.3 times (p < 0.05) and decreased PAP by 2.6 times (p < 0.05), while in uPA+/+ males, changes in PAI-I and PAP were opposite; in uPA-/- females, levels of all indicators increased by 1.6–2.1 times (p < 0.05), unlike uPA+/+ females. Among animals with melanoma only, changes in the levels of uPAR, PAI-I and PAP in the brain tissues in uPA-/- males differed from the group with CNP and from uPA+/+ males; in uPA+/+ females, levels of uPAR and PAP increased by 1.7 and 3.0 times (p < 0.05), and only PAP increased in uPA-/- females by 3.2 times (p < 0.05). Combination of CNP with melanoma in uPA-/- mice, regardless of their gender, down-regulated levels of uPAR and PAI-I on the average by 1.5 and 2.0 times, respectively (p < 0.05), and up-regulated PAP on the average by 2.2 times (p < 0.05) compared to the levels in animals with CNP; in uPA+/+ animals, similar decline of uPAR by 3.7 times (p < 0.05) was registered only in males, and an increase of PAI-I by 2.0 times (p < 0.05) was noted in all mice.Conclusion. Changes in the studied parameters in the brain tissue of urokinase gene-knockout animals in response to stress factors indicate the role of the brain urokinase system in the response to both CNP and melanoma growth, and the gender specificity of these changes may be another factor that conditions gender differences in the risk of occurrence and course of cutaneous melanoma.