Ml Of Saline Research Articles

Morphological characteristics of the cerebral cortex of a mini-pig under conditions of gene therapy after experimental stroke

Aim. To study the effectiveness of preventive gene therapy (within 2 days) and gene therapy in the acute phase (after 4 hours) of ischemic stroke in mini-pigs using an autologous leucoconcentrate (AutoLeuc) enriched with recombinant genes of vascular endothelial growth factor (VEGF165), glial cell line-derived neurotrophic factor (GDNF) and neural cell adhesion molecule 1 (NCAM1), as well as the migration of leukocytes transduced with a chimeric adenoviral vector serotype 5 with fiber 35 serotype (Ad5/F35) and the green fluorescent protein (GFP) genome into immune defense organs.Materials and methods. The experiment was conducted on 8-month-old Vietnamese lop-bellied mini-pigs (n=16). An ischemic stroke was created by occlusion of the distal branches of the left middle cerebral artery and the right common carotid artery. Genetically modified AutoLeuc was administered preventively intravenously 2 days before or in the acute phase 4 hours after stroke modelling; the control group was injected with 30 ml of saline solution. The morphology of the cerebral cortex was assessed using histological methods in the areas bordering the infarction and peri-infarction after 21 days. The migration of genetically modified Ad5/F35-GFP leukocytes into the brain, spleen, and submandibular lymph nodes was studied a week after stroke modelling.Results. In the peri-infarction zone, the content of pyknotic neurons in control animals was higher, while the number of capillaries was lower than in the gene therapy groups. In the latter, neurons had a typical morphology with preserved outgrowths; in the control group, the outgrowths were tortuous and fragmented. Fluorescence microscopy after injection of AutoLeuc with Ad5/F35-GFP revealed GFP-positive cells in the spleen and submandibular lymph nodes.Conclusion. 21 days after modeling a stroke in mini-pigs against the background of preventive gene therapy or gene therapy in the acute phase using VEGF165/GDNF/NCAM1 AutoLeuc, greater preservation of neurons and a higher density of capillaries in the peri-infarction zone of ischemic brain damage were established. Leukocytes with Ad5/ F35-GFP were found in the spleen and submandibular lymph nodes.

Effect of post-mating GnRH or TAK-683 (Kisspeptin analog) treatment on the reproductive performance and serum progesterone concentration in ewes during the non-breeding season

This study was designed to evaluate the effect of post-mating GnRH or TAK-683 (kisspeptin analog) treatment on serum progesterone (P4) concentrations and fertility parameters of ewes during the non-breeding season. One hundred and twenty multiparous, lactating Awassi ewes were used. For estrus synchronization, vaginal sponges containing 60 mg of medroxyprogesterone acetate (MAP) were inserted into the vagina and kept there for nine days. On the day of sponge removal (day 0), 500 IU of eCG and 75 µg of d-cloprostenol were given intramuscularly. Rams were introduced in the flock 24 h after sponge removal and kept with the ewes for 26 days, allowing a second mating period after synchronized estrus. On the fourth day after mating (day 6) the ewes mated by the rams between the 24th and 72nd hours (n = 105) were randomly divided into three groups. Control group (n = 36) received 1 ml of saline solution subcutaneously; GnRH group (n = 34) received 4 μg of buserelin acetate intramuscularly; TAK-683 group (n = 35) received 5 μg of TAK-683 acetate subcutaneously. Blood samples were taken to measure serum P4 concentrations (ng/ml) at sponge insertion and on the 6th, 9th, 12th, 15th, 19th, 35th, and 50th days after sponge removal. Pregnancy diagnosis was conducted via serum P4 concentration measurement on the 19th day and transrectal ultrasonography on days 35 and 50. The results showed that GnRH treatment (4.823 ± 0.238; 2.151 ± 0.272) increased (P<0.05) the mean P4 concentration (ng/ml) in pregnant and non-pregnant ewes during the first mating period compared to Control (3.708 ± 0.207; 1.515 ± 0.164), while TAK-683 treatment (4.457 ± 0.181; 1.967 ± 0.213) was not different between groups. GnRH (58.82 %) and TAK-683 (42.10 %) treatments increased the pregnancy rate in the second mating period in comparison to the Control group (17.39 %) (P<0.05). Cumulative pregnancy and fecundity rates were higher in GnRH (79.41 %; 120.58 %) and TAK-683 (68.57 %; 100.0 %) groups than in the Control group (17.39 %; 58.33 %) (P<0.05). There was no difference in late embryonic loss rate among the groups (P>0.05), whereas early fetal loss rate tended to be lower in the TAK-683 (0 %) and GnRH (0 %) groups than in the Control group (15.38 %) (P=0.07). The total pregnancy loss rate (from day 19 to day 50 post-mating) was lower in GnRH (0 %) and TAK-683 (0 %) groups relative to the Control group (21.42 %) (P<0.05). There were no statistical differences among the groups in terms of the first mating period pregnancy rate, twinning rate, and litter size (P>0.05). As a result, treatment with GnRH and TAK-683 on the fourth day after mating increased fecundity in lactating ewes outside the breeding season by preventing total pregnancy losses (from day 19 to day 50) and increasing second mating pregnancy rates.

Mitigating Acetaminophen-Induced Kidney Injury: The Protective Role of Grape Seed and Peanut Skin Extracts through the iNOS/CYP2E1 Pathway.

The rising number of acute kidney injury cases worldwide due to acetaminophen (APAP) emphasizes the critical need for effective prevention strategies to counteract APAP's detrimental effects. This study examined the kidney-protective capabilities of ethanolic extracts from grape seeds and peanut skins (GSEE and PSEE, respectively) in comparison with silymarin in rats that experienced an APAP overdose. The phenolic compounds in these extracts were measured by using high-performance liquid chromatography (HPLC). In the experiment, Sixty adult male albino rats were divided into five groups of 12. The Control group received 0.5 mL of saline via a gastric tube. Group II received acetaminophen (APAP, 640 mg/kg per day via a gastric tube) to induce renal injury, following Ucar et al. and Islam et al. Groups III, IV, and V received silymarin (50 mg/kg), grape seed extract (200 mg/kg), and peanut skin extract (200 mg/kg), respectively, along with 640 mg of APAP/kg per day for 21 days. Post APAP treatment, significant increases in serum urea and creatinine levels were noted, along with notable decreases in the percentage of body weight gain. Furthermore, there were increases in oxidative stress and inflammatory markers in the kidney tissues, including heightened mRNA expressions of renal iNOS and CYP2E1, which were confirmed through histological studies. The administration of GSEE, PSEE, and silymarin mitigated these adverse effects, likely due to their high phenolic content, which is recognized for its antioxidant and anti-inflammatory effects. GSEE, in particular, showed efficacy comparable to that of silymarin. Molecular docking studies revealed that APAP impeded critical enzymes essential for cellular antioxidant defense, whereas the bioactive compounds in the grape seed and peanut skin extracts effectively inhibited key enzymes and receptors involved in inflammation and oxidative stress. These findings suggest that GSEE and PSEE could serve as viable alternative treatments for kidney damage induced by APAP. Further research to isolate and identify these effective compounds is recommended.

Potential therapeutic properties of broccoli extract and soy isoflavones on improvement endometriosis and involved oxidative parameters.

In Endometriosis is a gynecological disorder characterized by the growth of endometrial tissue outside the uterine cavity that is associated with chronic pelvic pain and subfertility. The purpose of the study was to investigate the effect of broccoli extract (BE) alone and in combination with soy isoflavones (SI) on endometrial implants in female rat. In this study, endometriosis was induced surgically in 40 mature female rats. The rats were divided into 5 groups that were treated by oral gavage for 6weeks with 0.5 mL of saline 0.9 %/day (control group), BE (3,000 mg/kg/day), SI (50 mg/kg/day), BE/soy isoflavones (BE 3000 mg/kg/day+soy isoflavones 50 mg/kg/day) and diphereline as a standard medication (3 mg/kg) intramuscularly. At the end of treatments, the volume and histopathology of the endometrial implants were compared among the 5 groups. The serum levels of oxidative parameters including superoxide dismutase (SOD), malondialdehyde (MDA) and tumor necrosis factor alpha (TNF-α) were also compared between the groups. The volume of the implants significantly decreased in diphereline group (p=0.002). The histopathological grade of endometrial implants in BE/SI and diphereline group were significantly decreased compared to the control group (p=0.001). The serum levels of SOD in BE group were enhanced significantly in comparison to the control group (p=0.034). BE in combination with SI decreased the growth and histopathologic grades of transplanted endometrial implants. These herbal compounds may have the potential therapeutic effect to be used as an alternative medication for the treatment of endometriosis.

The modulatory effects of tyrosol and nano-tyrosol on anxiety-like behavior and emotional memory in streptozotocin-induced diabetic rats.

The effects of tyrosol and nano-tyrosol on the modulation of anxiety-like behavior and memory processes were evaluated in streptozotocin-induced diabetic rats. Male diabetic rats were orally treated with 1 ml of saline, nano-niosome, tyrosol, and nano-tyrosol (20 mg/dl) for 30 days. Anxiety-like behavior and memory process were evaluated by an elevated plus-maze (EPM) test-retest paradigm. The results showed that a single intraperitoneal (i.p.) administration of streptozotocin (50 mg/kg) raised blood glucose. While daily intragastric administration of tyrosol and nano-tyrosol reduced blood glucose. Induction of type II diabetes produced a distorted cellular arrangement whereas treatment with tyrosol and nano-tyrosol showed a typical cellular arrangement in the liver. Furthermore, induction of type II diabetes decreased %OAT (%open-arm time) but daily intragastric application of tyrosol (20 mg/dl) and nano-tyrosol (20 mg/dl) enhanced %OAT and %OAE (%open-arm entry) in the EPM when compared to the saline groups, showing anxiogenic- and anxiolytic-like effects, respectively. Also, induction of type II diabetes increased %OAT while daily intragastric administration of tyrosol (20 mg/dl) and nano-tyrosol (20 mg/dl) decreased %OAT and %OAE in the EPM in comparison to the saline groups, displaying impairment and improvement of emotional memory, respectively. Interestingly, nano-tyrosol exhibited the highest significant effect rather than tyrosol. Upon these results, we proposed the beneficial effects of tyrosol and nano-tyrosol on the modulation of anxiety-like behavior and memory processes in streptozotocin-induced diabetic rats.

Exploring developmental MeHg impact on extraembryonic and cardiac vessels and its effect on cardiomyocyte contractility.

The toxicity of methylmercury (MeHg) during embryonic development is a relevant issue that remains unclear and deserves investigation. In this sense, there is evidence that links the intake of contaminated food with cardiovascular pathologies in human adults and children. Thus, this study aimed to verify the impact of MeHg on the structure and integrity of extraembryonic and cardiac blood vessels and the contractile function of cardiomyocytes, also evaluating embryonic weight and the cardiosomatic index (CSI). Thus, chicken embryos, used as an experimental model, were exposed to a single dose of 0.1μg MeHg/50 μl saline at E1.5 and analyzed at E10. After exposure, an increase in the number of extraembryonic blood vessels and the veins of the cardiac tissue was observed. These increases were accompanied by a reduction in the content of VEGF and VCAM proteins related to vessel growth and adhesiveness. Together, these results were related to reduced nitrite (NOx) levels. Furthermore, MeHg reduces the number of sarcomeres and increases the content of cardiac troponin I (cTnI), a protein that regulates contraction. In general, exposure to MeHg affected the integrity of extraembryonic and cardiac vessels and the contractile function of cardiomyocytes, which had a systemic impact evidenced by the reduction in embryonic weight gain and CSI.

Does Fibromyalgia Affect the Structure of the Dentate Gyrus?

Abstract Background The hippocampus is considered an important component of the limbic system of the brain. It is involved in memory and functional diversities. The term hippocampus typically refers to the dentate gyrus and the hippocampus proper, the cornu ammonis (CA). Fibromyalgia is known to cause memory and cognitive function impairment and is characterized by causing chronic widespread muscular pain. Aim of the Work The aim of this work was to study the possible structural and morphometric changes in rats’ hippocampal dentate gyrus induced by repeated intramuscular injections of acidic saline in the left gastrocnemius and to compare it with the control. Material and Methods Fifty-four adult female albino Swiss rats were divided into three equal groups (18 rats each): control group (I), natural saline group (II) and acidic saline group (III). Group I received no medication, group II received two i.m. injections of 100 μl saline (pH 7.4) into the left gastrocnemius muscle at day 0 and 5 respectively under sedation with halothane (2-4%), while group III received two i.m. injections of 100 μl saline (pH 4) into the left gastrocnemius muscle at day 0 and 5 respectively under sedation with halothane (2-4%). Rats of each group were further subdivided into 3 subgroups (a, b, c) 6 rats each. Rats were sacrificed by decapitation at day 7, 14, 21 from the start of experiment and their brains were excised. Results Histological examination of H&amp;E-stained sections of groups I and II showed no difference and were both considered control groups. The dentate region was formed of three laminae: the molecular, granular and polymorphic. The granular cell layer is the principal layer separating the molecular and polymorphic layers and was formed of aggregation of granular cells. They were rounded to oval with large vesicular nuclei and .prominent nucleoli The examination of the H&amp;E-stained sections of rats’ dentate gyrus granular cells (DG) appeared darkly stained with pyknotic nuclei I group III as compared to the control. Glial cells slightly increased especially among molecular and polymorphic layers. Neurofibrillary tangles and vacuolations started to appear in silver-stained sections. The neurodegenerative findings started to be more notable in subgroup (b) then became more prevalent in subgroup(c) together with the appearance of clear areas among the sub granular zone. Morphometric studies detected non-significant difference in mean area percentage of astrocytes between group I and group II subgroups. The statistical analysis demonstrated high significant difference in mean area percentage of astrocytes between control groups and acidic saline groups. Evident gliosis was confirmed in group IIIc when the astrocytes percentage areas become very highly significantly increased compared to groups I and II. Conclusion In conclusion, repeated acidic saline injection induced dentate gyrus alterations obvious but minimal at week 1 and become intense at week 3 after the first injection. This effect ended with neuronal death and gliosis.

The role of mesenchymal stem cell therapy in ameliorating diabetes-induced ovarian damage in albino rats: Histological, biochemical and immunohistochemical studies

Background Diabetes mellitus (DM) is a chronic disease causing a variety of complications that can damage most organs, including the gonads. Diabetes was reported to be effectively treated using mesenchymal stem cells (MSCs). Objective The objective of the work was to demonstrate the therapeutic role of stem cells in cases of ovarian damage due to diabetes. Materials and methods Eighteen adult female albino rats were divided evenly into three groups. The control group received an injection of 1 ml of saline intraperitoneally (i.p.). The streptozotocin (STZ)-treated group: that was given 60 mg/kg per body weight (i.p.) of STZ to induce DM, which was slaughtered after 4 weeks. The STZ+stem cell-treated group that got STZ then was allowed to recover for 4 weeks and then underwent an intravenous injection of one million MSC before being sacrificed. Measurements of blood glucose, serum estradiol (E2), follicular-stimulating hormone (FSH), and luteinizing hormone (LH) were performed. Histological examination with hematoxylin and eosin, Masson stain, and immunohistochemical investigation for proliferating cell nuclear antigen (PCNA) were performed on the ovarian sections. Results and conclusion The STZ-induced DM group showed high glucose, FSH, and LH levels and decreased the E2 levels. Also, the inflammation, degeneration, and decreased PCNA immune expression of the ovary development in the STZ-induced DM group were observed. The treatment with the MScs reversed the effect of DM on glucose, FSH, LH, and E2. Additionally, the MScs improved the pathological alterations by increasing PCNA immune expression in the ovary. It was concluded that MSCs can effectively treat diabetes-induced ovarian damage.

Methanol extract of Abrus precatorius seeds on reproduction indices, hepatorenal profiles in female albino rats

BackgroundAbrus precatorious is a member of the leguminous family with characteristic red and black seeds. The roots, leaves and seeds of this plant are used for different medicinal purpose. It principally carries flavonoids, triterpene glycosides, abrin and alkaloids. The plant had been mentioned for neuromuscular effects, neuro-protective, abortifacient, antiepileptic, anti-viral, antimalarial, antifertility, nephroprotective, immunomodulator, immunostimulatory properties, antiinflammatory activity, antidiabetic, and many others. The aim of this study was to investigate the anti-fertility activity of the methanol extract of Abrus precatorius seeds (APS) in female albino rats administered 45 and 50 mg/kg BW through oral gavage for 30 days. An Abrus precatorius seed (APS) was extracted (1:1 w/v) using methanol to obtain the crude extract. Liquid-liquid fractionation was performed on the crude methanol extract using solvents of different polarity.MethodsEighteen female rats were divided into three groups each containing six animals was used for this experiment. Group one (control), received 1 ml of saline solution, group two received 45 mg/kg and group three received 50 mg/kg body weight by oral gavage daily for a period of 30 days. At the end of the study, Body weight, Organ Weight, Estrous cycle, Hormonal Concentrations, Oxidative Stress Markers, Enzymatic Activities, Biochemical indices (Liver and Kidney panel) and Hematological parameters were also evaluated.ResultsConsequent upon the administration of A.precatorius seed methanol extract 45 and 50 mg/kg BW into the female rats resulted in changes pertaining to Body weight, Estrous cycle, Hormonal Concentrations, Antioxidant Enzymes, Serum Biochemical indices and Hematological indices significantly (p < 0.05) compared to control rats. Initial body weights are significantly (p < 0.05) indicated, whereas final weights were decreased significantly (p < 0.05). Organ weight changes i.e. Ovary, Liver and Kidney weights were found to be significantly (p < 0.05) decreased in APS administered rats compared to control. Estrous cycle study i.e. time duration of Proestrus, Estrus, Metestrus and Diestrus (in days) was significantly prolonged. Hormonal Concentrations represented by FSH, LH, Prolactin, Estradiol were significantly (p < 0.05) decreased, whereas Progesterone levels were significantly (p < 0.05) increased in APS administered female rats compared to control group of rats. All the antioxidant enzyme assayed CAT, SOD, GPx, GR and GST were found to be significantly (p < 0.05) decreased, whereas MDA concentration was found to be increased significantly (p < 0.05). Enzymatic activities and biomolecules includes Cholesterol and Ascorbic acid contents were significantly increased (p < 0.05), but G6PDH and Δ5 − 3β-HSD activities were found to be significantly (p < 0.05) decreased in APS administered group of female rats compared to control group. Biochemical indices of liver tissue presented by Bilirubin, SGPT, SGOT, Albumin and Globulin were found to be significantly (p < 0.05) increased whereas Alkaline Phosphate and Total Protein contents were found to be significantly (p < 0.05) decreased. The biochemical indices of kidney, i.e. Urea and Creatinine were significantly (p < 0.05) increased and Uric acid and Calcium contents were found to be significantly (p < 0.05) decreased in APS administered female rats compared to control group of rats. Hematological parameters including RBC, Hb, ESR and Clotting Time were found to be significantly (p < 0.05) increased but WBC levels were significantly (p < 0.05) increased in APS administered female rats compared to control group of rats.ConclusionA. Precatorius seeds have anti-fertility effect on female rats.

Comparison of perineural and systemic dexamethasone use in impacted third molar surgeries in terms of anesthesia duration and postoperative complaints: a controlled, randomized observational study

BackgroundSurgical extraction of impacted third molars (ITM) often leads to postoperative discomfort including pain, swelling, and limited function. Steroids like dexamethasone (DXN) are commonly used in oral surgery to manage pain and inflammation. Various administration routes for DXN exist, including intravenous (IV), perineural (PN), and oral applications, each with its advantages. Previous studies have shown that adding DXN to local anesthetics can prolong anesthesia duration and reduce postoperative sequelae. However, comparative studies on IV and PN applications with inferior alveolar nerve block (IANB) of DXN in ITM surgeries are limited.MethodsThis controlled, randomized observational study involved patients undergoing Class II position B ITM extraction. Patients were divided into three groups. IANB (1.8 ml of articaine hydrochloride + 1 ml of saline) was performed 1 h after IV-DXN (4 mg/ml DXN) was administered to the IV group. DXN along with IANB (1.8 ml of articaine hydrochloride + 1 ml of 4 mg/ml DXN) was applied to the PN group. Only IANB (1.8 ml of articaine hydrochloride + 1 ml of saline) was applied to the control group. Anesthesia duration was assessed as primary outcomes. Anesthesia duration was evaluated using a vitalometer from the molars. Secondary outcomes included postoperative pain and edema measured on the 1st, 3rd, and 7th days after surgery. Pain was evaluated postoperatively by using a visual analog scale. A p-value < 0.05 was considered statistically significant.ResultsThe study included 45 patients with similar demographic characteristics across groups. IV application significantly prolonged anesthesia duration compared to the control group. (p = 0.049) Both IV and PN administration of DXN reduced postoperative edema at 3rd (p = 0.048) and 7th day (p = 0.01). Post-procedure pain reduction was significant in the IV group (p = 0.011). On the other hand, it was observed that the pain did not decrease in the PN group at 3rd and 7th days compared to the control and IV groups.ConclusionsPN and IV DXN administration prolonged anesthesia duration and reduced postoperative edema in ITM surgeries. However, PN DXN administration was associated with increased postoperative pain compared to IV DXN and control groups. Further studies comparing different doses and administration routes of DXN are needed to determine optimal strategies for managing postoperative discomfort in ITM surgeries.Trial registrationThis study was conducted at Ahmet Keleşoğlu Faculty of Dentistry with the permission of Karamanoğlu Mehmetbey University Faculty of Medicine Ethics Committee (#04-2022/101). Trial registration is also available at clinicaltrail.gov. (NCT06318013, 26/05/2024)

Intravenous magnesium sulfate improves orotracheal intubation conditions: A randomized clinical trial

BackgroundMuscle relaxation facilitates orotracheal intubation, but in some situations, neuromuscular blocking agents may be contraindicated. AimTo study the effect of intravenous magnesium sulfate perfusion before induction on orotracheal intubation conditions and the pressor response upon intubation. MethodsThis prospective, randomized, double-blind trial compared magnesium sulfate with saline solution as a placebo. The patients were randomized into two groups: the magnesium group (Group Mg+), which received 50 mg/kg of magnesium sulfate in 100 ml of saline 15 min before induction of anesthesia, and the saline group (Group Mg-), which received only 100 ml of saline 15 min before induction of anesthesia without magnesium sulfate. The induction in both groups was performed without neuromuscular blocking agents.Intubation conditions in both groups were compared using the Copenhagen consensus conference: ease of laryngoscopy, vocal cord position and/or movement, and response to intubation or cuff (cough or diaphragmatic movement). Intubation conditions were considered acceptable (excellent or good) or unacceptable (poor). Systolic, diastolic arterial pressures and heart rate were also recorded during the study. ResultsIn total, 76 patients (38 in each group) were included in this study. Clinically acceptable intubation conditions were significantly more observed in the magnesium group Mg + compared to the saline group Mg-: 36 patients (95 %) vs. 15 patients (39 %) (P < 0.001).Hemodynamic stability was more observed in the magnesium group, with a statistically significant lower heart rate throughout Mg + compared to the saline group. ConclusionAdministration of magnesium sulfate before induction of anesthesia significantly improved intubation conditions without the need for neuromuscular blocking agents.

Effects of intravenous inflammasome inhibitor (NuSepin) on suppression of proinflammatory cytokines release induced by cardiopulmonary bypass in swine model: a pilot study

The systemic inflammatory response syndrome can occur due to an inflammatory reaction to the release of cytokines, and it has been linked to the circulation of pro- and anti-inflammatory cytokines. The cardiopulmonary bypass (CPB) system is known to activate numerous inflammatory pathways. Applying CPB in large animals for an extended period may be useful as a controlled experimental model for systemic inflammatory responses. The authors hypothesized that 0.2 mg/kg NuSepin® would inhibit CBP-induced proinflammatory cytokine release, and attenuate CPB-induced vasoplegia. CPB was maintained for 2 h in 8 male Yorkshire pigs. Ten ml of saline was administered intravenously to the control group, while the study group received 10 ml of NuSepin® (0.2 mg/kg), before start of CPB. Blood samples were collected at four different time points to evaluating the level of cytokine (TNF-α, IL-1β, IL-6, IL-8) release during and after CBP. All vital signals were recorded as continuous waveforms using the vital recorder®. Our study demonstrated that IL-6 increased in both groups during CPB remained unchanged. However, in the Nusepin group, IL-6 levels rapidly decreased when CPB was stopped and the proinflammatory reaction subsided. Furthermore, the dose of norepinephrine required to maintain a mean pressure of 60 mmHg was also lower in the Nusepin group.

Pharmacologic blockade of nicotinic receptors in the suprachiasmatic nucleus increases ovarian atresia and inhibits follicular growth.

Reproduction in all mammalian species depends on the growth and maturation of ovarian follicles, that is, folliculogenesis. Follicular development can culminate with the rupture of mature follicles and the consequent expulsion of their oocytes (ovulation) or in atresia, characterized by the arrest of development and eventual degeneration. These processes are regulated by different neuroendocrine signals arising at different hypothalamic nuclei, including the suprachiasmatic nucleus (SCN). In the later, the activation of muscarinic receptors (mAChRs) and nicotinic receptors (nAChRs) by acetylcholine is essential for the regulation of the pre-ovulatory signals that stimulate the rupture of mature follicles. To evaluate the participation of the nAChRs in the SCN throughout the oestrous cycle in the regulation of the hypothalamic-pituitary-ovarian axis. For this purpose, 90-day-old adult female rats in metoestrus, dioestrus, proestrus or oestrus were microinjected into the left- or right-SCN with 0.3 μL of saline solution as vehicle or with 0.225 μg of mecamylamine (Mec), a non-selective antagonist of the nicotinic receptors, diluted in 0.3 μL of vehicle. The animals were sacrificed when they presented vaginal cornification, indicative of oestrus stage, and the effects of the unilateral pharmacological blockade of the nAChRs in the SCN on follicular development, ovulation and secretion of oestradiol and follicle-stimulating hormone (FSH) were evaluated. The microinjection of Mec decreased the serum levels of FSH, which resulted in a lower number of growing and healthy follicles and an increase in atresia. The higher percentage of atresia in pre-ovulatory follicles was related to a decrease in the number of ova shed and abnormalities in oestradiol secretion. We also detected asymmetric responses between the left and right treatments that depended on the stage of the oestrous cycle. The present results allow us to suggest that during all the stages of the oestrous cycle, cholinergic signals that act on the nAChRs in the SCN are pivotal to modulate the secretion of gonadotropins and hence the physiology of the ovaries. Further research is needed to determine if such signals are generated by the cholinergic neurons in the SCN or by cholinergic afferents to the SCN.

Glucocorticoids disrupt longitudinal advance of cortical bone basic multicellular units in the rabbit distal tibia

Glucocorticoids (GCs) are the leading cause of secondary osteoporosis. The emerging perspective, derived primarily from 2D histological study of trabecular bone, is that GC-induced bone loss arises through the uncoupling of bone formation and resorption at the level of the basic multicellular unit (BMU), which carries out bone remodeling. Here we explore the impact of GCs on cortical bone remodeling in the rabbit model. Based upon the rapid reduction of bone formation and initial elevation of resorption caused by GCs, we hypothesized that the rate of advance (longitudinal erosion rate; LER) of cortical BMUs would be increased. To test this hypothesis we divided 20 female New Zealand White rabbits into four experimental groups: ovariohysterectomy (OVH), glucocorticoid (GC), OVH + GC and SHAM controls (n = 5 animals each). Ten weeks post-surgery (OVH or sham), and two weeks after the initiation of dosing (daily subcutaneous injections of 1.5 mg/kg of methylprednisolone sodium succinate in the GC-treated groups and 1 ml of saline for the others), the right tibiae were scanned in vivo using Synchrotron Radiation (SR) in-line phase contrast micro-CT at the Canadian Light Source. After an additional 2 weeks of dosing, the rabbits were euthanized and ex vivo images were collected using desktop micro-CT. The datasets were co-registered in 3D and LER was calculated as the distance traversed by BMU cutting-cones in the 14-day interval between scans. Counter to our hypothesis, LER was greatly reduced in GC-treated rabbits. Mean LER was lower in GC (4.27 μm/d; p < 0.001) and OVH + GC (4.19 μm/d; p < 0.001), while similar in OVH (40.13 μm/d; p = 0.990), compared to SHAM (40.44 μm/d). This approximately 90 % reduction in LER with GCs was also associated with an overall disruption of BMU progression, with radial expansion of the remodeling space occurring in all directions. This unexpected outcome suggests that GCs do not simply uncouple formation and resorption within cortical BMUs and highlights the value of the time-lapsed 4D approach employed.

Comparative effects of 17% ethylenediaminetetraacetic acid and 9% etidronic acid applied with different irrigant activation techniques on the release of growth factors from dentin: in vitro study.

Growth factors embedded in the extracellular matrix of the dentin play an important role in the migration, proliferation, and differentiation of dental pulp stem cells in regenerative endodontics. In regenerative endodontic treatments, the type of irrigation solution used is crucial for the release of growth factors (GFs) from the dentin matrix. This study evaluated the effectiveness of different irrigant activation techniques (IAT) using two different chelating agents, 17% ethylenediaminetetraacetic acid (EDTA) and 9% etidronic acid (HEDP), in terms of their GF release. Seventy-two mandibular premolar teeth were prepared to simulate an open apex. The root fragments were irrigated with 20ml of 1.5% sodium hypochlorite and 20ml of saline solution. Eight root fragments were randomly separated for the control group, and the remaining 64 fragments were randomly separated into eight groups based on two different chelating agents (17% EDTA and 9% HEDP) and four different IAT ((conventional needle irrigation (CNI), passive ultrasonic irrigation (PUI), sonic activation with EDDY, and XP-endo Finisher (XPF)). TGF-β1, VEGF-A, BMP-7 and IGF-1 release levels were determined using an ELISA, and statistical analysis was performed using the Kolmogorov-Smirnov test, ANOVA, and the Tukey test (p < .05). Compared to the control group, the experimental groups showed significantly higher GF release when using EDTA or HEDP. Among the activation groups, the EDDY group triggered the highest GF release, and the CNI group triggered the lowest. IAT with EDTA and HEDP can increase GF release, with EDDY being the most effective IAT method. Using chelating agents with IAT may be beneficial in regenerative endodontic treatments.

#1983 In vitro iohexol adsorption in cartridges with mesoporous styrene-divinylbenzene sorbent

Abstract Background and Aims Patients with pre-existing chronic kidney disease (CKD) present the highest risk of developing contrast-associated acute kidney injury (CA-AKI). Iodinated contrast agents such as iohexol (molecular weight 821 Da) don't bind to proteins and are easily removed by hemodialysis filters. However, the session time necessary to remove approximately 75% of the injected iohexol load is around six hours. Due to lack of evidence, current guidelines do not recommend to start hemodialysis in CKD patients in conservative management after the administration of iodinated contrast media to prevent or mitigate CA-AKI. Still, a strong rationale supports the removal of iodinated contrast media because of its nephrotoxicity. In this study, we aimed to investigate the capability of a mesoporous styrene-divinylbenzene sorbent to bind iohexol in vitro. Method We used 1,000 mL of saline solution spiked with 40 mL of iohexol 350 mg/mL in a closed-loop configuration. Iohexol concentration was 14 mg/mL. The amount of the iodinated radiocontrast was proportional to the amount used in complex percutaneous coronary interventions (worst-case scenario), around 200 mL, in a 70 kg person with an estimated blood volume of 5,000 mL. The solution circulated through a HA380 mini-module cartridge filled with 75g of mesoporous styrene-divinylbenzene sorbent beads at 250 ml/min. The experiment was carried out twice with the same setup. Samples were collected at eight-time points (0, 5, 10, 15, 20, 30, 40, and 60 minutes) from the reservoir to measure iohexol concentration. The analysis of the samples was carried out in duplicate. The removal ratio (RR) was calculated considering the initial concentration (C(0)) and the variation of the concentration during the experiment (C(t), where t is the time point), as follows: $RR( \% ) = \frac{{C( 0 ) - C( t )}}{{C( 0 )}}100$ Results In the first experiment, the removal ratios after 5, 30, and 60 minutes were 23.4%, 45.8%, and 52.9%, respectively on average. In the second experiment, the reduction ratios after 5, 30, and 60 minutes were 23.4%, 45.5%, and 53.1%, respectively on average. After 30 minutes, the adsorption curved plateaued, indicating cartridge saturation (Fig. 2). Conclusion This in vitro model demonstrated that HA380 sorbent cartridge was effective in removing iohexol. The use of hemoadsorption as a strategy to remove iodinated contrast media warrants further investigation because of its clinical potential to reduce the exposure to nephrotoxic contrast agents and CA-AKI.

#860 The role of immunocorrection in the treatment of patients chronic pyelonephritis

Abstract Background and Aims Chronic pyelonephritis in the structure of extragenital pathology in pregnant women occupies a leading place due to the high prevalence, complicated course of pregnancy and childbirth. Chronic pyelonephritis (CP) triggers a cascade of reactions, primarily immune, at the systemic and local levels. With HP the role of the immune system in pregnant women is not only a pathological process occurring in the kidneys, but also takes into account a number of changes in the restructuring of the immune system of a pregnant woman. In relation with the above, the study of the role of the immune system of pregnant women with chronic pyelonephritis is of great interest. To study the effectiveness of immunocorrective therapy in the complex treatment of patients with chronic pyelonephritis. Method 43 pregnant women with CP were examined, the average age was 28.3 ± 0.2 years. The study included clinical research methods (survey, questionnaire, objective examination). Of the laboratory methods a general analysis of blood, urine, urine analysis according to Nechiporenko, tank. urine culture, biochemical blood tests and instrumental (ultrasound) research methods. Cellular immunity was studied by monoclonal antibodies, humoral immunity was studied by Mancini immunodiffusion. Among the surveyed women, the first group of upp was pregnant women (n = 14) receiving traditional therapy. The second group of 13 pregnant women with CP on the background of traditional therapy received ozone therapy (basic treatment + 400 ml of saline solution ozonated to a concentration of ozone in a liquid of 4 mg/l, intravenously, drip). The third group of 15 pregnant women with CP - with traditional therapy received ozone therapy and immune-regulating therapy with polyoxidonium. Results The characterization of clinical and laboratory data on the background of therapy showed significant shifts, as lower back pain decreased by 85.71%, 92.31% and 93.33%, respectively, in both the first and second and third groups. Indicators such as dysuria decreased by 92.85%, 92% and 93% (respectively in the first, second and third groups). Leukocyturia decreased significantly on the fifth day by 71.43%, 84.62%, 86.67% (in the first, second and third groups, respectively). Results indicators of eradication of pathogens after treatment in combination with traditional therapy with ozone and immune-correcting therapy, they were significantly better than in the ozone therapy group against the background of traditional therapy and in the traditional treatment group (86.6%, 84.6% and 64.28%, respectively). As a result of the study, significant decreases in cellular immunity indicators, in particular CD3, CD4, were revealed before the treatment of patients and CD8 and revealed a significant increase in IgA, IgM. After the treatment in the treatment group in combination significant increases in the number of CD3, CD4 and CD8 were revealed by traditional therapy with ozone- and immune-regulating therapy (by 76.1%, 64.68% and 58.3%, respectively), compared with the group of women who received only ozone therapy against the background of traditional therapy and in the group of traditional treatment (66.23%, 59.54%, 54.2% and 61.12%, 54.68%, 51.4%, respectively), and normalization of immunoglobulins A was noted and M. Conclusion Along with the improvement of clinical and laboratory data in CP normalization of cellular and humoral immunity indicators was noted in pregnant women. Additional assignment to traditional therapy for chronic pyelonephritis of pregnant women, ozone therapy and therapy for correction of the immune system is a more effective treatment method.

Aspects of Histopathological and Ultrastructural Retinal Changes in Chronic Exposure to Hydroxychloroquine.

Background and Objective: Hydroxychloroquine sulfate (HCQ) is a lysosomotropic agent administered in systemic lupus erythematosus and rheumatoid arthritis that has fewer toxic effects than chloroquine. However, HCQ may still be responsible for retinal toxicity. In this study, we observed structural changes in the retinas of experimental rats after prolonged exposure to HCQ. Matherials and Methods: We investigated several aspects regarding retinal changes, at both the histopathological and ultrastructural levels. We used 96 male albino Wistar rats distributed into four equal groups (n = 24 per group): the first three groups were treated with different doses of HCQ (50, 100, and 200 mg/kg HCQ, injected intraperitoneally in a single dose daily), and the last group (the control group, n = 24) was treated with saline solution administered in the same way (0.4 mL of saline solution). The treated groups received HCQ daily for 4 months, and every month, six animals from each group were sacrificed to assess retinal changes. The eyes were examined via optical (OM) and electronic microscopy (EM). Statistical analysis was deployed, and results regarding retinal morpho-photometry were acquired. Results: We observed structural retinal changes in both high and low doses of HCQ; while high doses determined a significant thinning of the retina, lower doses caused retinal thickening. Morphological retinal changes upon exposure to HCQ are believed to be caused by accumulated HCQ in lysosomes found in retinal ganglion cells and in the inner nuclear and photoreceptor cell layers. Such changes were most evident in the group receiving HCQ intraperitoneally in doses of 100 mg/kg for a longer period (4 months). Conclusions: The present study highlights histopathological and ultrastructural retinal changes induced by chronic HCQ administration, which were strongly connected to the dosage and period of exposure.

Methylene blue dose-dependently induces cutaneous inflammation and heat hyperalgesia in a novel rat model.

Methylene blue (MB) has been shown to reduce mortality and morbidity in vasoplegic patients after cardiac surgery. Though MB is considered to be safe, extravasation of MB leading to cutaneous toxicity has been reported. In this study, we sought to characterize MB-induced cutaneous toxicity and investigate the underlying mechanisms. To induce MB-induced cutaneous toxicity, we injected 64 adult male Sprague-Dawley rates with 200 µL saline (vehicle) or 1%, 0.1%, or 0.01% MB in the plantar hind paws. Paw swelling, skin histologic changes, and heat and mechanical hyperalgesia were measured. Injection of 1%, but not 0.1% or 0.01% MB, produced significant paw swelling compared to saline. Injection of 1% MB produced heat hyperalgesia but not mechanical hyperalgesia. Pain behaviors were unchanged following injections of 0.1% or 0.01% MB. Global transcriptomic analysis by RNAseq identified 117 differentially expressed genes (111 upregulated, 6 downregulated). Ingenuity Pathway Analysis showed an increased quantity of leukocytes, increased lipids, and decreased apoptosis of myeloid cells and phagocytes with activation of IL-1β and Fos as the two major regulatory hubs. qPCR showed a 16-fold increase in IL-6 mRNA. Thus, using a novel rat model of MB-induced cutaneous toxicity, we show that infiltration of 1% MB into cutaneous tissue causes a dose-dependent pro-inflammatory response, highlighting potential roles of IL-6, IL-1β, and Fos. Thus, anesthesiologists should administer dilute MB intravenously through peripheral venous catheters. Higher concentrations of MB (1%) should be administered through a central venous catheter to minimize the risk of cutaneous toxicity.

End-ischemic pharmacological cocktail treatment to mitigate rewarming/reperfusion injury

Increasing shortage of donor organs leads to the acceptance of less than optimal grafts for transplantation, up to and including organs donated after circulatory standstill of the donor.Therefore, protective strategies and pharmacological interventions destined to reduce ischemia induced tissue injury are considered a worthwhile focus of research.The present study evaluates the potential of a multidrug pharmacological approach as single flush at the end of static preservation to protect the liver from reperfusion injury.Livers were retrieved from male Wistar rats 20 min after cardiac standstill. The organs were cold stored for 18 h, flushed with 20 ml of saline, kept at room temperature for 20 min, and reperfused at 37 °C with oxygenated Williams E solution. In half of the cases, the flush solution was supplemented with a cocktail containing metformin, bucladesine and cyclosporin A.Upon reperfusion, treated livers disclosed a massive mitigation of hepatic release of alanine aminotransferase and aspartate aminotransferase, along with a significant approximately 50 % reduction of radical mediated lipid peroxidation, caspase activation and release of TNF-alpha.Even after preceding cold preservation, a pharmacological cocktail given as single flush is capable to mitigate manifestations of reperfusion injury in the present model.