Norepinephrine Concentrations Research Articles

ВЗАИМОСВЯЗЬ ПОКАЗАТЕЛЕЙ НАПРЯЖЕННОСТИ СИМПАТОАДРЕНАЛОВОЙ СИСТЕМЫ У ПАЦИЕНТОВ С ФИБРИЛЛЯЦИЕЙ ПРЕДСЕРДИЙ ДО И ПОСЛЕ ХИРУРГИЧЕСКОГО ЛЕЧЕНИЯ

Introduction. It is known that cardiovascular diseases, including cardiac arrhythmias, are accompanied by increased activation of the sympathetic-adrenal system (SAS). Hyperactivation of the SAS causes an increase in the level of circulating catecholamines - adrenaline and norepinephrine, resulting in desensitization of β-adrenergic receptors (β-AR), aggravation of myocardial contractile dysfunction and the development of arrhythmia. Indicators of SAS tension are β-adrenergic reactivity of erythrocyte membranes (β-ARM) and the level of circulating catecholamines. One of the most common arrhythmias is atrial fibrillation (AF), which leads to a decrease in the quality of life, disability and high mortality. Therefore, among patients with AF, it is important to study the functional viability of β-AR in different periods of the disease. Purpose: to evaluate β-ARM and the level of adrenaline, norepinephrine in patients with AF before and after surgical treatment. Materials and methods. The sample included 47 patients - 31 men and 16 women, the age in the sample was 51 (26; 77) years. All patients were diagnosed with AF based on ECG and 24-hour monitoring. Among the entire sample, 59.6% of patients had paroxysmal, 25.5% persistent, 14.9% long-term persistent forms of AF. All patients underwent surgical treatment of AF using radiofrequency or cryoablation. β-ARM was determined by the method of assessing changes in erythrocyte osmotic resistance as a result of blockade of β-AR in vitro with a selective β-adrenergic blocker (Β-ARM, AGAT, Russia). Plasma epinephrine and norepinephrine concentrations were determined by enzyme-linked immunosorbent assay using Adrenalin ELISA and Noradrenalin ELISA kits (Tecan IBL International, Germany). The parameters were studied in all patients before treatment, in 44 patients after 3 days, in 18 patients after 3 months and in 9 patients after 12 months after ablation. The significance of changes in β-ARM and catecholamine levels before and after ablation was assessed using the Wilcoxon test. The strength of the linear relationship between quantitative indicators was assessed using the Spearman rank correlation coefficient. Results. In the study sample, there were no statistically significant changes in β-ARM and adrenaline levels before and at different times after ablation. Thus, the level of β-ARM before surgery was 18.2 (12.6; 25.4), after 3 days - 22.3 (14.4; 26.6), after 3 months - 20.3 (9.3 ; 29.3), and 12 months after the operation - 27.7 (16.8; 36.7). The level of adrenaline before surgery, 3 days, 3 months and 12 months after surgery was 4.4 (3.7; 6.9), 4.7 (3.0; 6.4), 4.6 (2 .5; 6.6), 5.6 (9.1; 8.8). However, an increase in the level of norepinephrine was found 12 months after ablation compared with the values before treatment and 3 days after treatment by 1.7 and 1.8 times, respectively (p = 0.028). The level of norepinephrine before surgery, 3 days, 3 months and 12 months after surgery was 40.9 (18.7; 56.6), 38.6 (14.8; 49.6), 46.7 (25 .6; 57.3), 69.3 (65.1; 80.1). Before surgery, β-ARM and norepinephrine levels were linearly correlated (r = 0.335, p = 0.037), but after treatment, the linear correlation disappeared. At 3 and 12 months after ablation, there was a strong linear correlation between adrenaline and norepinephrine levels (r = 0.611, p = 0.012; r = 0.786, p = 0.036, respectively). Conclusion. In the sample of patients with AF, β-ARM and norepinephrine levels were linearly correlated before treatment, but after ablation, the correlation disappeared. At the same time, the values of β-ARM do not change statistically significantly, and the level of norepinephrine increases during a 12-month follow-up. Thus, 12 months after treatment in patients with AF, the activity of the CAS increases, but the functional viability of beta-AR does not change significantly.

Biomarkers associated with functional improvement after stroke rehabilitation: a systematic review and meta-analysis of randomized controlled trials.

This study aims to identify blood and cerebrospinal fluid biomarkers that are correlated to the functional improvement of stroke patients after rehabilitation therapy, and provide ideas for the treatment and evaluation of stroke patients. The PubMed, Web of Science, and Embase databases were searched for articles published in the English language, from inception to December 8, 2022. A total of 9,810 independent records generated 50 high-quality randomized controlled trials on 119 biomarkers. Among these records, 37 articles were included for the meta-analysis (with a total of 2,567 stroke patients), and 101 peripheral blood and cerebrospinal fluid biomarkers were included for the qualitative analysis. The quantitative analysis results revealed a moderate quality evidence that stroke rehabilitation significantly increased the level of brain-derived neurotrophic factor (BDNF) in serum. Furthermore, the low-quality evidence revealed that stroke rehabilitation significantly increased the concentration of serum noradrenaline (NE), peripheral blood superoxide dismutase (SOD), peripheral blood albumin (ALB), peripheral blood hemoglobin (HB), and peripheral blood catalase (CAT), but significantly decreased the concentration of serum endothelin (ET) and glutamate. In addition, the changes in concentration of these biomarkers were associated with significant improvements in post-stroke function. The serum BNDF suggests that this can be used as a biomarker for non-invasive brain stimulation (NIBS) therapy, and to predict the improvement of stroke patients. The concentration of serum BNDF, NE, ET and glutamate, and peripheral blood SOD, ALB, HB and CAT may suggest the function improvement of stroke patients.

Effects of the menstrual cycle on EPOC and fat oxidation after low-volume high-intensity interval training.

Low-volume high-intensity interval training (HIIT) for weight loss has become prevalent in recent years, with increased excess post-exercise oxygen consumption (EPOC) as the mechanism. However, the influence of the menstrual cycle on EPOC and fat oxidation following low-volume HIIT is unclear. This study aimed to investigate the effect of the menstrual cycle on the increase in EPOC and fat oxidation after low-volume HIIT. Twelve eumenorrheic women participated during their early follicular and luteal phases. On each experimental day, they performed low-volume HIIT comprising fifteen repeated 8 s sprint cycling tests with 12 s rests, for 5 min. Expired gas samples were collected before and every 60 min until 180 min post-exercise. EPOC was defined as the increase in oxygen consumption from the resting state, and the total EPOC and fat oxidation were calculated from the total time of each measurement. Blood samples for serum estradiol, progesterone, free fatty acids, blood glucose, lactate, and plasma noradrenaline were collected and assessed before immediately after, and at 180 min post-exercise and were assessed. Serum estradiol and progesterone were significantly higher in the luteal phase than the follicular phase (P<0.01 for both). No significant differences in total EPOC and fat oxidation were found between the menstrual phases. Serum free fatty acid, blood glucose, lactate, and plasma noradrenaline concentrations were not affected by the menstrual cycle. These results suggest that the menstrual cycle does not affect the increase in EPOC or fat oxidation after low-volume HIIT.

Electroacupuncture intervention improves lipid metabolism and promotes browning of white adipose tissue by activating AMPK/Sirt1 pathway and up-regulating Nrg4 content in middle-aged and aged obese rats

To investigate the mechanism of electroacupuncture （EA） in promoting the browning of white adipose tissue in middle-aged and aged obese rats induced by high fat by regulating AMP-activated protein kinase （AMPK） /silence-information regulatory factor 1 （Sirt1） pathway and neuregulin 4 （Nrg4）. Twenty-four male SD rats were randomized into blank control, model and EA groups （n=8 per group）. The obesity model was established by feeding the rats with high-fat diet for 6 weeks. For the EA group, EA （2 Hz/15 Hz, 1.5 mA） was applied to "Guanyuan" （CV4） and bilateral "Shenshu" （BL23）, "Fenglong" （ST40） and "Tianshu" （ST25） for 20 min, once a day, 5 days a week for 6 weeks. Rats of the blank control and model groups were also restrained for 20 min. The body mass and food intake were measured every week, and the Lee's index, epididymal fat, perirenal fat and brown adipose tissue were weighed. The contents of serum total cholesterol （TC）, triglyceride （TG）, high density lipoprotein cholesterol （HDL-C）, low density lipoprotein cholesterol （LDL-C） and norepinephrine （NE） were determined by ELISA. H.E. staining was used to observe the morphological changes of white and brown adipose tissue. The mRNA expression levels of mitochondrial uncoupling protein 1 （UCP1）, peroxisome proliferator activated receptor γ co-activator 1α （PGC-1α）, PR-domain protein 16 （PRDM16）, peroxisome proliferator activated receptor γ （PPARγ） and Nrg4 in the adipose tissue were detected by quantitative real time PCR, and the protein expression levels of Nrg4, AMPKα, Sirt1 and interleukin-6 （IL-6） in the white and brown adipose tissue were detected by Western blot. Compared with the blank control group, the body mass, food intake, the Lee's index, epididymal fat and perirenal fat mass, and serum TG, TC and LDL-C contents and the expression level of IL-6 protein were significantly increased （P<0.01, P<0.05, P<0.001）, and the brown adipose mass, serum HDL-C and NE contents, the expression levels of UCP1, PGC-1α, PRDM16, PPARγ and Nrg4 mRNAs, and the protein expression levels of AMPKα, Sirt1 and Nrg4 proteins in both white and brown adipose tissues were significantly decreased in the model group （P<0.01, P<0.05）. After EA intervention, the increased levels of body mass, food intake, Lee's index, epididymal fat and perirenal fat mass, serum TG, TC and LDL-C contents, and the expression of IL-6 protein, and the decreased levels of brown adipose mass, serum HDL-C and NE contents, expression levels of UCP1, PGC-1α, PRDM16, PPARγ and Nrg4 mRNAs, and those of AMPKα, Sirt1 and Nrg4 proteins in both white and brown adipose tissues were apparently reversed（P<0.05, P<0.01, P<0.001）. H.E. staining showed an increase of the volume and content of intracellular vacuoles of both white and brown adipose tissues, disordered arrangement of cells with vague boundary in the model group, which was relatively milder including a decrease of volume and content of vacuoles of both white and brown adipose, neat arrangement of cells with clear boundary. EA intervention can improve lipid metabolism and promote white adipose tissue browning in middle-aged and aged obese rats, which is possibly associated with its functions in activating AMPK/Sirt1 signaling pathway and up-regulating the level of Nrg4.

Electroacupuncture intervention improves lipid metabolism and promotes browning of white adipose tissue by activating AMPK/Sirt1 pathway and up-regulating Nrg4 content in middle-aged and aged obese rats

To investigate the mechanism of electroacupuncture （EA） in promoting the browning of white adipose tissue in middle-aged and aged obese rats induced by high fat by regulating AMP-activated protein kinase （AMPK） /silence-information regulatory factor 1 （Sirt1） pathway and neuregulin 4 （Nrg4）. Twenty-four male SD rats were randomized into blank control, model and EA groups （n=8 per group）. The obesity model was established by feeding the rats with high-fat diet for 6 weeks. For the EA group, EA （2 Hz/15 Hz, 1.5 mA） was applied to "Guanyuan" （CV4） and bilateral "Shenshu" （BL23）, "Fenglong" （ST40） and "Tianshu" （ST25） for 20 min, once a day, 5 days a week for 6 weeks. Rats of the blank control and model groups were also restrained for 20 min. The body mass and food intake were measured every week, and the Lee's index, epididymal fat, perirenal fat and brown adipose tissue were weighed. The contents of serum total cholesterol （TC）, triglyceride （TG）, high density lipoprotein cholesterol （HDL-C）, low density lipoprotein cholesterol （LDL-C） and norepinephrine （NE） were determined by ELISA. H.E. staining was used to observe the morphological changes of white and brown adipose tissue. The mRNA expression levels of mitochondrial uncoupling protein 1 （UCP1）, peroxisome proliferator activated receptor γ co-activator 1α （PGC-1α）, PR-domain protein 16 （PRDM16）, peroxisome proliferator activated receptor γ （PPARγ） and Nrg4 in the adipose tissue were detected by quantitative real time PCR, and the protein expression levels of Nrg4, AMPKα, Sirt1 and interleukin-6 （IL-6） in the white and brown adipose tissue were detected by Western blot. Compared with the blank control group, the body mass, food intake, the Lee's index, epididymal fat and perirenal fat mass, and serum TG, TC and LDL-C contents and the expression level of IL-6 protein were significantly increased （P<0.01, P<0.05, P<0.001）, and the brown adipose mass, serum HDL-C and NE contents, the expression levels of UCP1, PGC-1α, PRDM16, PPARγ and Nrg4 mRNAs, and the protein expression levels of AMPKα, Sirt1 and Nrg4 proteins in both white and brown adipose tissues were significantly decreased in the model group （P<0.01, P<0.05）. After EA intervention, the increased levels of body mass, food intake, Lee's index, epididymal fat and perirenal fat mass, serum TG, TC and LDL-C contents, and the expression of IL-6 protein, and the decreased levels of brown adipose mass, serum HDL-C and NE contents, expression levels of UCP1, PGC-1α, PRDM16, PPARγ and Nrg4 mRNAs, and those of AMPKα, Sirt1 and Nrg4 proteins in both white and brown adipose tissues were apparently reversed（P<0.05, P<0.01, P<0.001）. H.E. staining showed an increase of the volume and content of intracellular vacuoles of both white and brown adipose tissues, disordered arrangement of cells with vague boundary in the model group, which was relatively milder including a decrease of volume and content of vacuoles of both white and brown adipose, neat arrangement of cells with clear boundary. EA intervention can improve lipid metabolism and promote white adipose tissue browning in middle-aged and aged obese rats, which is possibly associated with its functions in activating AMPK/Sirt1 signaling pathway and up-regulating the level of Nrg4.

Establishment of a Rat Model of Hypotension Induced by Reserpine

Objective To determine the optimal dosage and intervention duration of reserpine to establish a rat model of hypotension.Methods According to the body weight and systolic blood pressure (SBP),60 male Wistar rats were assigned to six groups (n=10),including a control group and five observation groups with different doses.The control group was administrated with 10 ml/kg 0.5% sodium carboxymethyl cellulose solution,and the observation groups with 0.016,0.032,0.064,0.128,and 0.160 mg/kg reserpine suspensions,respectively.All the groups were administrated by gavage twice a day,and the body weights of rats were monitored daily.SBP and heart rate (HR) were measured before modeling and 1-6 weeks after administration.After 6 weeks of administration,the blood samples of inner canthus were collected.The levels of lactate dehydrogenase (LDH),creatine kinase MB isoenzyme (CK-MB),alanine aminotransferase,aspartate aminotransferase (AST),serum creatinine,and blood urea nitrogen (BUN) were measured by an autoanalyzer.Three rats in each group were randomly selected for observation of the changes in SBP after drug withdrawal and the rest rats were sacrificed for measurement of the levels of norepinephrine and dopamine in the brain.Results Compared with the control group,different doses of reserpine lowered the SBP of rats (F=28.492,P<0.001).The decline in SBP increased in a concentration-dependent manner.SBP reached the lowest value after 1 week,rose slightly later,and was stable after 3 weeks of administration.There was no significant difference in SBP between 0.016 mg/kg reserpine group and the control group after the 5th week (P>0.05).The SBP levels of rats in 0.032,0.064,0.128,and 0.160 mg/kg reserpine groups showed no significant difference between each other (P=0.204) and were lower than that in the control group (all P<0.001).One week after drug withdrawal,the SBP of rats in the observation groups rose to the baseline level and remained stable.HR showed similar changes among groups,first increasing and then decreasing.There was no significant difference in HR among different groups at the same time point (F=0.922,P=0.475).Compared with the control group,reserpine of different doses reduced the norepinephrine content in the hippocampus (all P<0.001),and 0.128 mg/kg (P=0.045) and 0.160 mg/kg (P=0.042) reserpine lowered the dopamine level in the striatum,which showed no significant differences between different reserpine groups(P=0.343,P=0.301).The levels of LDH,CK-MB,and BUN in the serum increased with the increase in reserpine concentration,and the levels of LDH (P=0.001),CK-MB (P=0.020),AST (P=0.007),and BUN (P=0.001) in the 0.160 mg/kg reserpine group were significantly different from those in the control group.Conclusions The rat model of hypotension can be induced by gavage with reserpine.The gavage with reserpine at a dose of 0.032 mg/kg,twice a day for three consecutive weeks is the optimal scheme for the modeling.After the model establishment,continuous administration is essential to maintain the hypotension.

Unravelling Impaired Hypoalgesia at Rest and in Response to Exercise in Patients with Chronic Whiplash-Associated Disorders: Effects of a Single Administration of Selective Serotonin Reuptake Inhibitor versus Selective Norepinephrine Reuptake Inhibitor.

(1) Background: Noradrenaline and serotonin have modulatory roles in pain signaling and in exercise-induced hypoalgesia. Patients with chronic whiplash-associated disorders often show impaired exercise-induced hypoalgesia. Therefore, this study aimed to examine the isolated effect of activating serotonergic or noradrenergic descending pathways on hypoalgesia at rest and in response to exercise in patients with chronic WAD by using respectively a single dose of a selective serotonin reuptake inhibitor (SSRI) and a selective norepinephrine reuptake inhibitor (NRI). (2) Methods: Twenty-five people with chronic WAD participated in this double-blind randomized controlled crossover experiment. Serotonin and noradrenaline concentrations were modulated by the oral ingestion of a single dose of citalopram (i.e., SSRI) or atomoxetine (i.e., SNRI). Quantitative sensory testing (including pressure pain thresholds and conditioned pain modulation) was measured before and after exercise in combination with no medication (1), atomoxetine (2), or citalopram (3) at three different test days. (3) Results: Random-intercept linear mixed models analysis was used to analyze pain outcomes (i.e., pain at rest and exercise-induced hypoalgesia) before and after exercise over the three conditions in patients with chronic WAD. No differences in pain at rest were found between the three conditions before exercise. The effect of exercise on pain outcome measures was not influenced by medication intake. The occupational status of the participants had a significant influence on the effect of exercise and medication on pain outcomes (p < 0.05). Patients working full-time had some positive effect of atomoxetine on pain facilitation (p < 0.05). Unemployed patients had some negative effect of citalopram on pain tolerance and experienced exercise-induced hypoalgesia (p < 0.05). (4) Conclusions: A single dose of citalopram or atomoxetine did not result in changes in hypoalgesia at rest and in response to exercise. These results do not support the use of SSRI or selective NRI to overcome impaired hypoalgesia at rest or in response to exercise in people with chronic WAD. Effect of exercise and medication on pain in patients with chronic WAD is influenced by the occupational status.

Effect of acupuncture-moxibustion stimulation of combined "Biao-Ben" acupoints on autonomic nervous activity and related factors in rats with irritable bowel syndrome diarrhea

To observe the effect of acupuncture and moxibustion of the combined "Biao-Ben" acupoints on autonomic nervous function and related factors in rats with irritable bowel syndrome diarrhea (IBS-D). Female SD rats were randomly divided into normal control, IBS-D model, "Biao-Ben" acupoint combination, and conventional acupoint combination groups, with 10 rats in each group. The IBS-D model was established by repeated chronic stress stimulation (water or food deprivation, painful tail pinching, exposure to a 43 ℃ environment, forced swimming in 4 ℃ water, day-night inversion and horizontal vibration) for 28 d, and followed by acute restraint stress (wrapping of shoulders, forelimbs and trunk) for 1 h and gavage of senna fluid, once daily for 28 d. For rats of the "Biao-Ben" acupoint combination group, acupuncture and moxibustion were applied to "Guanyuan" (CV4), bilateral "Zusanli" (ST36), and "Neiguan" (PC6), and for rats of the conventional acupoint combination group, acupuncture and moxibustion were applied to bilateral ST36, "Tianshu"(ST25), and "Shangjuxu"(ST37). Both acupuncture and suspension moxibustion treatment were conduced for 15 min, once daily for 21 days. The fecal water content was calculated, and the spontaneous activity behaviors (total distance of crossing and the number of squares crossed in 5 min) were evaluated by open-field tests. The abdominal withdrawal reflex (AWR) was detected. H.E. staining was used to observe the histopathological changes of colon tissue. The domains of heart rate variability (HRV) including low frequency (LF), high frequency (HF), and LF/HF were analyzed using an electrophysiological recorder. The contents of calcitonin gene-related peptide (CGRP) in serum and noradrenaline (NE) in plasma were detected using ELISA. The expression level and immunoactivity of 5-hydroxytryptamine (5-HT) in colon tissues were detected by Western blot and immunofluorescence histochemistry, separately. Compared with the normal group, the model group had a striking increase in fecal water content, AWR scores at 20, 40, 60, and 80 mm Hg, NE and CGRP contents, LF, LF/HF, and 5-HT protein expression and immunoactivity (P<0.01), and an obvious decrease in the total distance of crossing, the number of squares crossed, and HF of HRV (P<0.01). In comparison with the model group, the fecal water content, AWR scores at 20, 40, 60, and 80 mm Hg, NE and CGRP contents, LF, LF/HF, and 5-HT protein expression and immunoactivity were significantly decreased (P<0.01,P<0.05), while the total distance of cros-sing, number of squares crossed, HF of HRV were considerably increased (P<0.01, P<0.05) in both "Biao-Ben" acupoint combination and conventional acupoint combination groups. The effects of the "Biao-Ben" acupoint combination were apparently superior to those of conventional acupoint combination in down-regulating the fecal water content, AWR score at 20, 40, 60 and 80 mm Hg, NE and CGRP contents, LF, LF/HF, and 5-HT expression and immunoactivity, and in increasing the number of squares crossed, and HF of HRV (P<0.01, P<0.05). HE staining showed no pathological changes in colonic mucosa in all groups. Acupuncture and moxibustion stimulation of combined "Biao-Ben" acupoints can effectively improve the symptoms (spontaneous activities, visceral hypersensitivity) of IBS-D model rats, which may be related to its functions in regulating autonomic nervous activities, and down-regulating blood NE and CGRP contents and colonic 5-HT protein expression and immunoactivity, and the effects of "Biao-Ben" acupoint combination are superior to those of conventional acupoint combination.

Methylphenidate induces a different response in the dorsal raphe as compared to ventral tegmental area and locus coeruleus: behavioral and concomitant neuronal recordings in adult rats.

Methylphenidate (MPD) is a psychostimulant used to treat attention deficit hyperactivity disorder. MPD exerts its neurocognitive effects through increasing concentrations of dopamine (DA), norepinephrine (NE), and serotonin (5-HT) in the neuronal synapse. This study recorded from adult freely behaving rats a total of 1170 neurons, 403 from the ventral tegmental area (VTA), 409 from locus coeruleus (LC), and 356 from dorsal raphe (DR) nucleus, which are the main sources of DA, NE, and 5-HT to the mesocorticolimbic circuitry, respectively. Electrophysiological and behavioral activities were recorded simultaneously following acute and repetitive (chronic) saline or 0.6, 2.5, or 10.0mg/kg MPD. The uniqueness of this study is the evaluation of neuronal activity based on the behavioral response to chronic MPD. Animals received daily saline or MPD administration on experimental days 1-6 (ED1-6), followed by a 3-day wash-out period, and then MPD rechallenge on ED10. Each chronic MPD dose elicits behavioral sensitization in some animals, while in others, behavioral tolerance. Neuronal excitation following chronic MPD was observed in brains areas of animals exhibiting behavioral sensitization, while neuronal attenuation following chronic MPD was observed in those animals expressing behavioral tolerance. DR neuronal activity was most affected in response to acute and chronic MPD administration and responded differently compared to the neurons recorded from VTA and LC neurons at all doses. This suggests that although not directly related, DR and 5-HT are involved in the acute and chronic effects of MPD in adult rats, but exhibit a different role in response to MPD.

(008) Chronic High Fat Diet Decreases Vaginal Smooth Muscle Tone but does not Affect Pelvic Floor Skeletal Muscle in Mice

Abstract Introduction Pelvic organ prolapse (POP) is a disease that affects the physical and mental health of women, as well as their quality of life. POP, regardless of severity, can be a direct cause of female sexual dysfunction (FSD) by producing pain during intercourse or a general reluctance to engage in sexual activity. The largest risk factors for POP include parity, obesity, and advanced age. Obesity, a modifiable risk factor, is believed to increase intra-abdominal pressure and weaken the pelvic floor muscles and fascia. Studies have shown that obesity and a high fat diet (HFD) can weaken skeletal muscle by increasing collagen deposition and inducing muscle apoptosis. Objectives There is paucity in the literature of animal studies that have specifically investigated the effect of obesity on the support structures of the pelvic floor and how it contributes to POP. The objective of this study is to examine the structure and function of the vagina and pelvic floor in female mice fed a chronic HFD. Methods Adult female C57/Blk6N mice (10 weeks, n=32) were fed a control (10% kcal fat) or HFD (45% kcal fat) for 24 weeks. Ex vivo external vaginal sphincter contraction to increasing concentrations of adrenergic agonist norepinephrine and relaxation to nitric oxide donor DEA NONOate and electric field stimulation (EFS) was measured. Iliococcygeus (IC) muscles of the pelvic floor were isolated and set to optimal length and tetanic contractions were performed at increasing frequencies to asses absolute and specific force. Fatigue was measured following repetitive stimulations over 5 minutes. IC muscle fiber types were characterized with anti-myosin antibodies and cross-sectional area (CSA) of each fiber type was measured. Results Vaginal sphincter adrenergic-mediated contraction was a significantly decreased in HFD-fed mice (Fig 1A; p&amp;lt;0.05). The vaginal sphincter smooth muscle relaxation in response to a nitric oxide donor was greater in the HFD mice (Fig 1B; p&amp;lt;0.05). Neurogenic-mediated EFS vaginal relaxations were also enhanced in the HFD mice (p&amp;lt;0.05). Surprisingly, chronic HFD did not impact iliococcygeus muscle specific force, absolute force, or muscle fatigue (Fig 1C). Furthermore, HFD did not affect the CSA or population of type I, IIa, IIX, or IIb iliococcygeus muscle fibers. Conclusions HFD-induced obesity in mice leads to pelvic floor weakness by enhancing relaxation and decreasing tone of the vaginal smooth muscle. Interestingly, the function and composition of pelvic skeletal muscle was unaffected by HFD. As the vagina serves as a primary support structure to the pelvic organs, lack in muscle contractility and increase in relaxation following HFD suggests that obesity-induced POP may play a direct role in FSD. Disclosure

Effect of "Tiaoshen Tongluo" acupuncture on nerve function injury, muscle tension and neurotransmitters through Nrf2/ROS pathway in spastic rats after stroke

To observe the effect of "Tiaoshen Tongluo" acupuncture (TTA) at "Dingzhongxian" (MS5) and right "Dingpangxian" (MS8) on neurological injury, muscle tension and neurotransmitters through nuclear transcription factor E2 related factor 2 (Nrf2)/reactive oxygen species (ROS) signaling pathway in spastic rats after stroke, so as to explore its mechanisms underlying relief of post-stroke spasm (PSS). A total of 90 male SD rats were randomly divided into 6 groups, i.e. sham operation, PSS model, medication, non-acupoint acupuncture, TTA, TTA+ML385 groups, with 15 rats in each group. The PSS model was established by middle cerebral artery occlusion. After modeling, rats of the medication group were treated by gavage of baclofen (0.4 mg/kg), once daily for 7 days. For rats of the non-acupoint acupuncture group, the spot about 10 mm above the iliac crest and below the armpit of the affected side was needled, and for those of the TTA group and TTA+ML385 group, EA stimulation (1 mA, 2 Hz/15 Hz) was applied to MS5 and right MS8 for 10 min, once daily for 7 consecutive days. Intraperitoneal injection of ML385 ［ a specific nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor, 30 mg/kg］ was given to rats of the TTA+ML385 group before TTA was performed. The rats' neurological deficit score (0-4 points) was evaluated by referring to Zea Longa's methods and the muscular spasm degree of the quadriceps femoris of the left hindlimb (0-4 points) assessed by using Ashworth scale (MAS). The muscular tension of the left quadriceps femoris was measured by using a tension sensor, and Hoffman (H)-reflex response and M and H waves of electromyogram of the muscle between the metatarsals of the left foot were measured using an electrophysiological recorder. The cerebral infarction volume was measured after 2,3,5-triphenyltetrazolium chloride (TTC) staining. The contents of γ-aminobutyric acid (GABA), glycine (Gly), glutamic acid (Glu) and aspartic acid (Asp) of the right cortical infarct area were detected by using high performance capillary electrophoresis, and the contents of 5-hydroxytryptamine (5-HT), dopamine (DA) and norepinephrine (NE) were detected by fluorescence spectrophoto-metry, as well as the level of ROS in the right cerebral cortical infarction tissues was detected by dihydroethidium staining. The expression levels of Nrf2 and heme oxygenase-1 (HO-1) proteins in the infarcted cerebral area were detected using Western blot. Compared with the sham operation group, the neurological deficit score, MAS score, percentage of cerebral infarction volume, Hmax/Mmax ratio, contents of Glu and Asp and ROS level were significantly increased (P<0.001), whereas the muscle tone, stimulation threshold for inducing H-reflex, GABA, Gly, 5-HT, DA and NE contents, cerebral Nrf2 and HO-1 protein expression levels were apparently decreased (P<0.001) in the model group. In comparison with the model group, the neurological deficit score, MAS score, percentage of cerebral infarction volume, Hmax/Mmax ratio, contents of Glu, Asp and ROS levels were decreased (P<0.001), and the muscle tone, stimulation threshold for inducing H-reflex, GABA, Gly, 5-HT, DA and NE contents, Nrf2 and HO-1 protein expressions were increased (P<0.001, P<0.01) in both the medication and TTA groups. No significant differences were found between the non-acupoint group and model group, and between the medication and TTA groups in all the indexes mentioned above (P>0.05). After administration of ML385, the effects of TTA in reducing neurological deficit score, MAS score, Hmax/Mmax, percentage of cerebral infarct volume, Glu, Asp, ROS, and up-regulating H-reflex threshold, GABA, Gly, 5-HT, DA, NE, Nrf2 and HO-1 levels were eliminated (P<0.001，P<0.05,P<0.01). TTA can improve neurological behavior and muscle spasm in rats with PSS, which may be associated with its functions in regulating the levels of neurotransmitters in the cortical infarcted area by activating the Nrf2/ROS signaling pathway.

Angiotensin-(1−7) improves tail skin heat loss and increases the survival of rats with polymicrobial sepsis

Sepsis is a serious syndrome, characterized by the excessive release of inflammatory mediators and thermoregulatory changes, being fever the most common sign. However, despite the importance of Angiotensin (Ang)-(1−7) in controlling the inflammation, the role of the peptide in the febrile response and mortality in animals submitted to experimental model of sepsis is still not clear. In this way, we evaluate the effect of continuous infusion of Ang-(1−7) in inflammatory response, thermoregulation and in mortality of Wistar male rats submitted to colonic ligation puncture (CLP). Before CLP surgery, the infusion pumps (Ang-(1−7), 1.5 mg/mL or saline) were inserted into the abdominal cavity and maintained for 24 h. CLP rats showed a febrile response starting from 3 h after and persisted until the 24th hour of experiment. Continuous treatment with Ang-(1−7) attenuated the febrile response and reestablished the euthermia 11 h after CLP, until the end of experiment, which coincided with an increased heat loss index (HLI). This effect was associated with a decrease in production of pro-inflammatory mediators in liver, white adipose tissue (WAT) and hypothalamus. Moreover, an increase in norepinephrine (NE) content in interscapular brown adipose tissue (iBAT) was observed in CLP animals, which was attenuated with treatment with Ang-(1−7), and decreased mortality in CLP animals treated with Ang-(1−7). Taken together, the present study demonstrates that continuous infusion treatment with Ang-(1−7) can promote a global anti-inflammatory effect, reestablishing the tail skin heat loss as a key thermo-effector function, resulting in an increased survival of animals submitted to experimental sepsis.

On the degree of effectiveness of surgical aids for mechanical jaundice

Relevance. The problem of mechanical jaundice is the most urgent in abdominal surgery. This is primarily because the frequency of this pathology is steadily increasing, and the mortality rate does not have a distinct downward trend. Surgical tactics in case of mechanical jaundice remains multivariate, and a differentiated approach is needed for treating this contingent of patients, which is decisive in improving the results of surgical aids.The aim of the study. To improve the results of treatment of patients with mechanical jaundice of calculous etiology.Materials and methods. The analysis of the results of treatment and specially conducted studies in 554 patients with mechanical jaundice of calculous etiology was carried out. The patients underwent ultrasound, CT, and MRI. The concentration of adrenaline, norepinephrine, dopamine, 11-oxycorticosteroids, histamine, and serotonin, as well as toxic metabolic products were determined. The state of the autonomic nervous system was assessed by determining the autonomic index and mathematical analysis of the heart rate using variational pulsometry.Results. All the patients were operated on. Priority was given to stage-by-stage surgical tactics combining endoscopic decompression of the biliary tract with endoscopic papillosphincterotomy in the early stages of hospitalization and open surgical treatment after cupping the clinic of mechanical jaundice. There was a 3-fold increase in the number of endoscopic interventions. The mortality rate for the entire period decreased by 2.1 times. The reason for the results obtained is the degree of trauma of the operation. The parameters of the musculoskeletal, autonomic nervous, and sympathoadrenal systems were determined in patients divided into three groups. In the first group, primary open surgery on the biliary tract was performed, in the second — endoscopic papillosphincterotomy and abdominal surgery on the biliary tract, in the third — only endoscopic papillosphincterotomy. Endoscopic papillosphincterotomy had the most sparing effect on the functioning of the cardiovascular system and normalization of vegetative homeostasis. The volume of the surgical aid for mechanical jaundice affects the content of the sympathoadrenal system and biologically active substances in the blood plasma. When performing abdominal operations, compared with minimally invasive ones, the concentration of adrenaline, norepinephrine, serotonin, histamine, and 11-oxycorticosteroids in the blood increased by 1.5–2.5 times.Conclusion. The effectiveness and priority of minimally invasive operations for treating mechanical jaundice have been confirmed. The use of endoscopic resolution of biliary hypertension makes it possible to reduce the mortality rate against the background of a steady trend toward normalization of the indicators of the sympathoadrenal system and variational pulsometry.

Eucalyptol prevents pilocarpine-induced seizure and neuronal damage in mice, through the cholinergic, monoaminergic and antioxidant pathways

Current antiepileptic drugs can inhibit seizure occurrence but are not effective in preventing its onset. Moreover, they produce several side effects, which may impact the efficacy of the treatment. In addition, it has been stimulating the prospection of new molecules isolated from aromatic plants, with potential anticonvulsant and neuroprotective activities and less side effects. This study aimed to evaluate, though behavioural and neurochemical methodologies, the anticonvulsant, and neuroprotective effects of eucalyptol on mice subjected to the pilocarpine-induced seizure model. Eucalyptol (100, 200 and 400 mg/kg, p.o.) was administered in mice prior to pilocarpine (350 mg/kg, i.p.) and the following behavioural parameters were assessed: Latency to First Seizure (LFS), Seizure Intensity (SI) and Latency to Death (LD). In addition, an oxotremorine-induced tremors test was performed to evaluate the cholinergic system involvement on the eucalyptol effects. Neurochemical tests were also performed, including determination of hippocampal (HC) concentration of thiobarbituric acid reactive substances (TBARS) and nitrite/nitrate and striatal (ST) concentration of noradrenaline, dopamine, and serotonin. Lastly, histopathological, and morphometric hippocampal analysis were conducted. Eucalyptol increased the latency to first seizure and latency to death, inhibited oxotremorine-induced tremors, decreased hippocampal TBARS and nitrite/nitrate overproduction, increased striatal noradrenaline and dopamine levels and prevented hippocampal neurodegeneration. These results demonstrate the potential anticonvulsant, neuroprotective and antioxidant effects of eucalyptol, probably through a conjunction of mechanisms including muscarinic cholinergic antagonism, oxidative stress mitigation and the monoaminergic system modulation, which appears to effectively control the seizure onset.

Investigations on some indicators in the blood of cattle with orosthenic activity tongue rolling.

Hematological studies were carried out in cattle with tongue-rolling hyperactivity in order to establish the etiopathogenetic mechanisms of this behavioral disorder. Cattle were divided into two groups: Group 1 (n = 12), control animals that did not demonstrate the tongue-rolling orosthenic activity, and Group 2 (n = 12), cattle that manifested this stereotyped behavior. Blood samples were collected from all cattle twice at 10-day intervals from Vena jugularis for analysis of red blood cell counts, hemoglobin (HGB) content, mean corpuscular volume (MCV), mean corpuscular HGB (MCH), mean corpuscular HGB concentration (MCHC), hematocrit (HCT), white blood cell counts (WBC), and differential white cell counts [lymphocytes (LYM), monocytes (MON), and granulocytes (GRAs)]. Some blood biochemical constituents were assayed: calcium, inorganic phosphorus, magnesium, plasma total protein, blood glucose, total bilirubin, urea, creatinine, chlorides, cholesterol, triglycerides, and albumin, as well as the enzyme activities of aspartate amino-transferase (AST), alanine amino-transferase (ALT), alkaline phosphatase (ALP), gamma-glutamyltransferase (γ-GT), lactate dehydrogenase (LDH), and creatine kinase (CK). Blood concentrations of the hormones adrenaline, noradrenaline, cortisol, adrenocorticotropic hormone (ACTH), dopamine, serotonin, free thyroxine, thyroid-stimulating hormone, and trace elements manganese, copper, and iron were determined. It was found out that the cattle with tongue-rolling hyperactivity did not demonstrate changes in red blood picture (HGB, HCT, MCV, MCH, MCHC), white blood picture (WBC, LYM, MON, GRA), blood biochemistry (Ca, P, Mg, TP, Glu, TB, urea, creatine, Cl, Chole, TG, and Alb), AST, ALT, ALP, γ-GT, LDH, and CK activities, Cu, and Fe levels. In the study, increased concentrations of adrenaline, noradrenaline, cortisol, ACTH, and serotonin were established. The data demonstrating increased levels of adrenaline, noradrenaline, cortisol, and ACTH confirmed the etiological role of environmental stressors in the emergence of oral hyperactivity. Increased serotonin concentrations in the blood suggest that cattle with the stereotype are probably "happy" with tongue rolling. The lack of changes in blood trace elements manganese, copper, and iron allowed excluding their deficiency as a probable cause in the etiology of the disorder.

Effect of Gabapentin-Fluoxetine Derivative GBP1F in a Murine Model of Depression, Anxiety and Cognition.

Gabapentin is a commonly prescribed antiepileptic agent for seizures, which is also used for pain and addiction management. Due to growing evidence of its abuse liability, there has been an incentive to synthesise potentially useful gabapentin derivatives devoid of adverse effects. A gabapentin adduct with a fluoxetine moiety, GBP1F, was assessed for any sedative, cognitive, anxiolytic, or antidepressant-like actions in murine behavioral models. Selected groups of mice were used for each behavioral paradigm, and the effect of GBP1F (5, 10, and 15 mg/kg) was assessed using spontaneous locomotor activity, the tail suspension test, elevated plus maze test, and the Y maze test models. Immediately following behavioral experiments, postmortem striatal and hippocampal tissues were evaluated for the effect of GBP1F on concentrations of dopamine, DOPAC, HVA, serotonin, 5-HIAA, vitamin C, and noradrenaline using high performance liquid chromatography with electrochemical detection. GBP1F induced a mild suppression of locomotor activity, ameliorated anxiety and depression-like behavior, did not alter cognitive behavior, and raised serotonin and 5-HIAA concentrations in the hippocampus and striatum. GBP1F also positively enhanced dopamine and vitamin C tissue levels in the striatum. Thus, GBP1F represents a compound with anxiolytic- and antidepressant-like effects though further studies are warranted at the molecular level to focus on the precise mechanism(s) of action.

THE CLINICAL CHARACTERISTICS OF PATIENTS WITH NORMOTENSION IN PHEOCHROMOCYTOMAS AND PARAGANGLIOMAS

Objective: The predominant symptom of pheochromocytomas and paragangliomas (PPGLs) is variable hypertension (HTN); however, not all patients with PPGLs develop HTN. Studies of the clinical characteristics of normotensive patients and which patients are more likely to develop HTN in PPGLs are rare. This study was performed to identify the clinical features to better understand this clinical entity. Design and method: All consecutive adult patients with PPGLs were retrospectively enrolled from January 2018 to June 2020. The clinical characteristics of the patients were investigated. Multivariate logistic regression analysis was used to identify risk factors and receiver operating characteristic curves were used to evaluate the diagnostic performance of the variables. Results: There were 203 patients in the analysis, including 115 patients with normotension. Fewer patients with normotensive PPGLs had diabetes (p&lt;0.001) and clinical symptoms (p&lt;0.001). The 24-h urinary epinephrine (p = 0.002) and 24-h urinary norepinephrine (24hU-NE) concentrations (p&lt;0.001) were lower, and tumor diameter (p&lt;0.001) was smaller in patients with normotensive PPGLs. Multivariate logistic regression analysis showed that diabetes and 24hU-NE concentration were independent and negative risk factors for normotensive PPGLs. The area under the curve of 24hU-NE concentration and diabetes for predicting normotensive PPGLs was 0.788 and 0.634, respectively. Combining diabetes and 24hU-NE concentration into one model, the area under the curve of the model for predicting normotensive PPGLs was 0.817. Conclusions: Normal blood pressure couldn’t exclude PPGLs. Identifying the clinical characteristics of normotensive and hypertensive patients in PPGLs is helpful for risk stratification and individualized assessment and treatment.

HIGH SALT DIET ALTERS CEREBROVASCULAR RESISTANCE IN RESPONSE TO ORTHOSTATIC TEST IN HEALTHY YOUNG INDIVIDUALS

Objective: High salt (HS) diet has been shown to impairs vascular reactivity in animal models, by increasing vasoconstriction to different stimuli. Response to orthostasis occurs via activation of baroreceptor reflex leading to increase in total peripheral resistance that prevents significant fall of arterial blood pressure (ABP) while standing up, aiming to preserve cerebral perfusion pressure. We recently showed that HS diet physiologically suppresses sympathetic arm of autonomic control in healthy humans (Stupin et al. Nutrients 2021; 13:1731. https://doi.org/10.3390/nu13051731). Present study aimed to assess the effect of HS diet on cerebral vasoreactivity in response to orthostatic load. Design and method: Twenty-seven young healthy participants (F:21, M:6, age range 19–24) consumed 7-day low-salt (LS) diet (&lt;2.3 g NaCl /day) and afterwards 7-day HS diet (&gt;11.2 g NaCl/day). In response to orthostatic test cerebral and systemic hemodynamic parameters, arterial blood pressure, heart rate variability and heart rate, and were recorded simultaneously with transcranial Doppler ultrasound and The Task Force® Monitor. Results: Subjects maintained normotension during HS diet. Diastolic blood pressure, as well as mean cerebral blood flow (CBF) and the velocity time integral decreased, while cerebral pulsatility index increased after standing up following both dietary protocols. However, while no change was observed in total peripheral resistance, cerebrovascular resistance significantly increased in response to orthostasis only after HS diet. Baroreflex sensitivity (BRS), sympathetic neural control, serum plasma renin activity and aldosterone, and urine noradrenaline and vanillylmandelic acid concentration was significantly decreased in HS diet. Conclusions: Results demonstrated that following 7-day HS diet, enhanced cerebral autoregulation preserves CBF and is manifested as increased cerebral vascular resistance, in conditions with blunted BRS and sympathetic activity. The study has been published in Eur J Appl Physiol. 2023 Jan 4. doi: 10.1007/s00421-022-05124-w. Online ahead of print.

Effects of chronic exposure to cold stress on ovarian functions in prepubertal rats.

Ovarian functions are modulated by the hypothalamus-pituitary-ovary axis and neural signals. Stress modifies the activity of the sympathetic nervous system. In adult female rats, cold stress results in higher noradrenergic and steroidogenic activity of the ovary, anovulation and the presence of ovarian cysts; however, it is unknown whether this response occurs in prepubertal rats. The purpose of this study was to analyse the effects of cold stress initiated in the prepubertal stage of female rats on ovarian function. Female rats 24 days old were exposed to three, five or eight weeks of cold stress. Autopsies were performed at the end of each stress period. The parameters analysed were the number of ova shed by ovulating animals; the number of ovulating animals; the serum concentrations of progesterone, testosterone, and oestradiol; and the ovarian concentrations of norepinephrine and 3-methoxy-4-hydroxyphenyl-glycol. Our results show that chronic cold stress applied to prepubertal rats did not modify the number of ovulating animals, the total number of ova shed, or progesterone and testosterone concentrations in any of the periods analysed. Oestradiol concentration was lower in the animals exposed to five or eight weeks of stress. The ovarian norepinephrine concentration was higher in the animals exposed to three weeks of stress and was lower at eight weeks of stress. No changes in ovarian morphology were observed. Our data suggest that the changes in noradrenergic activity resulting from chronic cold stress experienced in the prepubertal stage do not modify ovarian architecture or affect the ovulatory response in adulthood.

Norepinephrine as the Intrinsic Contributor to Contact Lens-Induced Pseudomonas aeruginosa Keratitis.

Contact lens wear (CLW) is one of the leading risk factors for Pseudomonas aeruginosa keratitis (PAK). However, the intrinsic factors that contribute to the high susceptibility to keratitis during CLW remain to be elucidated. CLW over an extended period can elevate corneal norepinephrine (NE) concentration. In this study, we investigated the role of NE in promoting PAK. We constructed an injury-induced PAK model and a CLW-induced PAK model to confirm the impact of NE during corneal infection. Pharmacological blockage of NE and gene knockdown mouse were used to investigate the downstream effector of NE. RNA sequencing was performed to explore the cellular alterations during NE treatment. Non-parametric Mann-Whitney U test or Kruskal-Wallis test were used to ascertain the significance (P < 0.05). Supplementation of NE led to PAK even without artificial corneal injury during CLW. The effect was mediated by the β2-adrenergic receptor (β2-AR) in the corneal epithelium. The β2-AR blockage by the NE antagonist ICI118,551 (ICI) or by deleting of its encoding gene Adrb2 significantly alleviated infection during CLW. Conversely, β2-AR activation compromised the integrity of the epithelium and significantly increased the cortical plaque marker ezrin. Transcriptome analysis identified that the protective effect of ICI on the keratitis was mediated by dual-specificity phosphatases. Suramin, a Dusp5 antagonist, abrogated the protective effect of ICI. These data reveal a new mechanism by which NE acts as an intrinsic factor that promotes CLW-induced PAK and provide novel therapeutic targets for treating keratitis by targeting NE-β2-AR.