Intraperitoneal Saline Research Articles

Low pressure pneumoperitoneum and intraperitoneal infusion of normal saline for reducing shoulder tip pain after gynecologic laparoscopy: randomized controlled trial

Background: To estimate the effectiveness of combined low-pressure pneumoperitoneum (8mmHg) and intraperitoneal normal saline infusion on reducing the incidence and severity of postoperative shoulder tip and upper abdominal pain.Methods: A prospective randomized controlled study was carried out in Aswan University Hospital, Aswan, Egypt. Author included patients undergoing laparoscopic surgery in the laparoscopy unit either diagnostic or operative. They were randomized into two groups: (intervention group) which compromised of 47 patients who underwent low pressure pneumoperitoneum plus intraperitoneal saline infusion and (control group) which included 47 patients underwent standard pressure pneumoperitoneum. The primary outcome of the study is the difference in the mean shoulder tip and upper abdominal pain score postoperatively between the two groups.Results: There was significant reduction in the shoulder tip pain and upper abdominal pain 1, 4, 8, 12 and 24 hours post-operatively shown by visual analogue scale pain scores in the intervention group compared to control (p=0.000). Additionally, there was a significant reduction in the incidence of nausea and vomiting in the intervention group than control group (p=0.000 and p=0.007 respectively). There was no significant difference regards abdominal distention, time of resumption of intestinal peristalsis, operation duration and post-operative hospital stay between the two groups.Conclusions: This study clearly depicts that combined low pressure pneumoperitoneum and intraperitoneal saline infusion is an easy, safe and inexpensive method that significantly reduces the post laparoscopic shoulder tip pain and upper abdominal pain.

Does dexmedetomidine prevent colistin nephrotoxicity?

BackgroundIn this study, we aimed to investigate the effect of dexmedetomidine on colistin nephrotoxicity in rats. MethodsThirty-two Wistar albino rats were allocated into four groups. Intraperitoneal (ip) saline at 1mL.kg−1 was administered to the control group and 10mg.kg−1 ip colistin was given to the colistin group. In the DEX10 group 10mcg.kg−1 dexmedetomidine ip was given 20min before the injection of 10mg.kg−1 ip colistin. In the DEX20 group ip 20mcg.kg−1 dexmedetomidine was injected 20min before the administration of 10mg.kg−1 ip colistin. These treatments were continued twice a day for seven days. Samples were taken on the eighth day. BUN, Cr, KIM-1, TAS, and TOS were examined in blood samples and caspase-3 was examined in kidney tissue samples. ResultsThe values for BUN, Cr and TOS were significantly higher in the colistin group than in the control group. BUN, Cr and TOS changes in the DEX10 and DEX20 groups were not significant compared with the control group but they were significantly lower compared with the colistin group. TAS values in the DEX10 group were significantly lower than in the control group. Apoptotic activity was significantly higher in the colistin group compared with the control group, but there was no significant difference in terms of caspase-3 staining activity when DEX10 and DEX20 groups were compared with the control group. ConclusionOxidative damage and apoptosis played roles in colistin nephrotoxicity, and colistin nephrotoxicity could be prevented by treatment with dexmedetomidine.

Intra-Peritoneal normal saline installation minimize post laparoscopic cholecystectomy shoulder pain in patient admitted to diwaniya teaching hospital

Intra-Peritoneal normal saline installation minimize post laparoscopic cholecystectomy shoulder pain in patient admitted to diwaniya teaching hospital

Efficacy of clarithromycin as a protective agent in the methotrexate-induced pulmonary fibrosis model.

IntroductionMethotrexate is a cytotoxic agent used in leukemia, and several other cancer types and at lower doses in auto-inflammatory diseases such as rheumatoid arthritis, ankylosing spondylitis and psoriasis. Macrolide antibiotics are effective against gram-positive and Gram-negative bacteria. They have anti-inflammatory activities as well. Clarithromycin is a macrolide with anti-inflammatory activity through blockage of the p38 MAPK signal cascade, which is involved in methotrexate-induced pulmonary toxicity.AimIn this study, the efficacy of clarithromycin in protecting against pulmonary fibrosis was investigated in the rat model for methotrexate-induced pulmonary fibrosis.Material and methodsA total of 30 female rats were divided into three groups. Group I was administered intraperitoneal and intragastric saline; group II was administered oral 3 mg/kg methotrexate; and group III was administered oral 3 mg/kg methotrexate + intraperitoneal 200 mg/kg clarithromycin for 28 days. Histopathological analyses of the lung tissues were performed under light microscopy.ResultsNormal histopathological changes were observed in the control group. Pulmonary fibrosis was significantly higher in the methotrexate group than in the other groups (p < 0.005).ConclusionsClarithromycin was shown to be effective in protecting against methotrexate-induced pulmonary fibrosis; further studies should be performed to determine the dosage and safety.

N-acetylcysteine attenuates the cuprizone-induced behavioral changes and oligodendrocyte loss in male C57BL/7 mice via its anti-inflammation actions.

Previous animal studies have linked white matter damage to certain schizophrenia-like behaviors in cuprizone (CPZ)-exposed mouse. Mitochondrial dysfunction, oxidative stress, neuroinflammation, and oligodendrocyte loss coexist in the brain of such mice. The aim of this study was to examine effects of the antioxidant N-acetylcysteine (NAC) on CPZ-induced behavioral changes and concurrent oligodendrocyte loss, oxidative stress, and neuroinflammation in these animals. Male C57BL/6 mice were given intraperitoneal saline or NAC at doses of 100, 200, and 400 mg/kg/day for 2 weeks; animals were fed a CPZ-containing diet (0.2%, w/w) during days 5-14. During days 15-17, the mice were examined in open-field, social recognition, and Y-maze tests (1 test per day). Six mice in each group were then used for biochemical and enzyme-linked immunosorbent assay analyses, while the remaining animals were used for immunohistochemical and immunofluorescence staining. The mice exposed to CPZ for 10 days showed significantly lower spontaneous alternation in the Y-maze, lower activity of total superoxide dismutase, and glutathione peroxidase, but higher levels of malondialdehyde in the cerebral cortex and hippocampus, elevated concentrations of interleukin-1β and tumor necrosis factor-α in the brain regions mentioned above and caudate putamen, and a decreased number of mature oligodendrocytes, but increased number of microglia in all the brain regions examined. These changes, however, were not seen or effectively alleviated in NAC-treated mice at all three doses. These results demonstrate that NAC protected mature oligodendrocytes against the toxic effects of CPZ, likely via its antioxidant and anti-inflammatory actions.

Could erythropoietin reduce the ovarian damage of cisplatin in female rats?

The aim of this study is to investigate whether erythropoietin (EPO) can reduce the ovarian damage of cisplatin or not. Thirty, female, Wistar-Albino rats were used in the study. Control group (N = 10): Intraperitoneal saline infusion, Cisplatin group (N = 10): Intraperitoneal 7 mg/kg cisplatin, Cisplatin + EPO group (N = 10): Intraperitoneal 7 mg/kg cisplatin and subcutaneous 200 IU/kg/day EPO. Serum AMH concentrations were measured by enzyme-linked immunosorbent assay kit of AMH. Follicular counts were evaluated according to mean diameter of the follicles. Ovarian damage; including follicular cell degeneration, vascular congestion, hemorrhage, and inflammation was scored histologically using a graduated scale. Posttreatment AMH levels of cisplatin group were significantly lower than control and cisplatin + EPO groups. In cisplatin group, there was a significant decrement in posttreatment AMH level compared to pretreatment AMH level. The total damage score of cisplatin group was significantly higher than scores of control and cisplatin + EPO groups. The mean primordial follicle counts of control and cisplatin + EPO groups were significantly higher than that of cisplatin group (p = .007 and p = .003). The results of this study revealed that EPO administration to cisplatin chemotherapy could ameliorate the ovarian damage. Erythropoietin administration to chemotherapeutic agents might suggest to protect ovarian failure and infertility.

Impairment of Hematopoietic Precursor Cell Activation during the Granulopoietic Response to Bacteremia in Mice with Chronic-Plus-Binge Alcohol Administration.

Alcohol abuse impairs immune defense. To study the effect of chronic-plus-binge alcohol exposure on the granulopoietic response, acute alcohol intoxication (intraperitoneal injection of 5 g alcohol/kg body weight) was introduced to mice chronically fed on the Lieber-DeCarli low-fat liquid alcohol diet for 5 weeks. Bacteremia was induced by intravenous injection of Escherichia coli Bacteremia caused a remarkable increase in marrow lin- c-kit+ Sca-1+ cells. Activation of cell proliferation supported the increase in marrow lin- c-kit+ Sca-1+ cells. Alcohol administration inhibited this activation of lin- c-kit+ Sca-1+ cells. The bone marrow of pair-fed control mice receiving intraperitoneal saline stored a large number of mature granulocytes expressing a high level of Gr1 (Gr1hi cells). The proportion of Gr1hi cells and the total number of Gr1+ cells were markedly reduced in the bone marrow, along with an increase in the ratio of Gr1+ granulocytes in peripheral white blood cells following bacteremia. E. coli infection stimulated proliferation of granulopoietic precursor cells, resulting in a marked increase in the ratio of immature Gr1lo cells in the bone marrow. Alcohol administration itself triggered marrow release of Gr1+ cells, resulting in reduction of the marrow granulocyte reserve with an elevation of granulocytes in the circulation. Alcohol also impaired activation of granulopoietic precursor proliferation following bacteremia. Alcohol disrupted lipopolysaccharide (LPS)-TLR4-ERK1/2-cyclin D1 signaling and inhibited upregulation of Sca-1 and C/EBPβ expression by lineage-negative marrow cells in response to bacteremia. These results indicate that chronic-plus-binge alcohol exposure inhibits the granulopoietic response by disrupting key cell signaling for hematopoietic precursor cell activation and commitment to granulocyte lineage development.

Profilactic role of simvastatin and mitomycin C in tracheal stenosis after tracheal damage: Study in rats

ObjectivesWe aimed to investigate the prophylactic effect simvastatin of and mitomycin C (MMC) on laryngeal and tracheal stenosis in tracheotomised rats by histopathological evaluation of laryngotracheal segment. Randomized prospective single-blind. Material and methodStandard vertical tracheotomy was performed on 24 rats. Then the animals were randomly divided into three groups as A, B and C. In group A 0.4 mg/day once daily mitomycin C was injected to the paratracheal region for 14 days. In group B daily 30 mg/kg/day simvastatin was given via gavage to rats for 14 days. In group C 2 cc/day intraperitoneal saline given to rats and the created control group by 14 days follow up. After 10 days, tracheal cannulas were removed. Three weeks later, all animals were euthanized and trachea specimens were harvested. The present study investigates the effects of MMC and Simvastatin on fibrosis, inflammation, stenosis index and tracheal wall thickness in a tracheal injury model. ResultsThe difference between the groups in terms of degree of inflammation scores was statistically insignificant (P = 0,187). Differences between the groups were found to be insignificant in terms of the preventionof fibrosis (P = 0,993). There was no significant difference between groups in terms of stenosis index (P = 0.645). In terms of wall thickness, control, simvastatin and mitomycin C groups were statistically different (p = 0.038). The difference between post-hoc test results was between Mitomycin C and control groups (p = 0.036).Maximum wall thickness in the MMC group (0,299 mm) was significantly lower compared to the control group (0,382 mm)(P < 0,0001). Maximum wall thickness was statistically lower in the simvastatin (0.324 mm) group compared with the control group (0.382 mm) (P < 0.0001). There was no statistically significant difference between the simvastatin group (0,198 mm) and control group (0,200 mm) with respect to minimum wall thickness (P = 0.982). Minimum wall thickness was significantly lower in the mitomycin-C group (0,160 mm) comparison to the control group (0,200 mm) (P < 0.0001). ConclusionIt was detected that the simvastatin and MMC is not effective in preventing the tracheal stenosis, inflammation and fibrosis formation.

Study of the protective effect of dexamethasone on cisplatin-induced ototoxicity in rats.

To evaluate the ability of dexamethasone to protect against cisplatin (CDDP)-induced ototoxicity. Male Wistar rats were divided into the following three groups: 1) Control (C): 6 animals received intraperitoneal (IP) saline solution, 8 ml/kg/day for four days; 2) C + CDDP: 11 animals received 8 ml/kg/day of IP saline and, 90 min after saline administration, 8 mg/kg/day of IP CDDP for four days; and 3) DEXA15 + CDDP: 11 animals received IP dexamethasone 15 mg/kg/day and, 90 min after dexamethasone administration, received 8 mg/kg/day of IP CDDP for four days. It was found that dexamethasone did not protect against weight loss in CDDP-exposed animals. The mortality rate was comparable with that previously reported in the literature. The auditory threshold of animals in the DEXA15 + CDDP group was not significantly altered after exposure to CDDP. The stria vascularis of animals in the DEXA15 + CDDP group was partially preserved after CDDP exposure. Dexamethasone at the dose of 15 mg/kg/day partially protected against CDDP-induced ototoxicity, based on functional evaluation by brainstem evoked response audiontry (BERA) and morphological evaluation by optical microscopy. However, dexamethasone did not protect against systemic toxicity.

Histological and immunohistochemical study on the potential toxicity of sliver nanoparticles on the structure of the spleen in adult male albino rats

Introduction: Silver nanoparticles (AgNPs) remain one of the controversial research areas regarding their distribution, dissolution and their toxicity to biological systems.Aim of the work: To evaluate the potential toxicity of different doses of AgNPs on the spleen of adult Albino rats.Materials and Methods: Forty adult male albino rats were divided into two main groups; control group: included ten rats that received intraperitoneal saline solution, daily, and experimental group: included thirty rats that received intraperitoneal AgNPs dissolved in saline solution, and were equally subdivided into three subgroups. Subgroup IIa received AgNPs (100 mg/kg/d), subgroup IIb received AgNPs (500 mg/kg/d) and subgroup IIc received AgNPs (1000 mg/kg/d). After 28 days, rats were euthanized and the spleen specimens were prepared for light and electron microscope studies.Results: Spleen of AgNPs-treated rats revealed dose-dependent structural changes in the form of reduced sized white pulp follicles with depletion of lymphocytes, degeneration and apoptosis. Nanoparticles were seen in both white and red pulps. There was a highly statistical significant increase in the optical density of caspase-3 in subgroups IIa, IIb and IIc when compared to that of the control group.Conclusions: AgNPs induced structural alterations in the spleen with evidence of inflammatory response and oxidative stress. These effects were dose-dependent.

EPR Oximetry of Cetuximab-Treated Head-and-Neck Tumours in a Mouse Model

Head and neck squamous cell carcinoma (HNSCC) tumours are associated with high mortality despite advances in therapy. The monoclonal antibody cetuximab (Erbitux®) has been approved for the treatment of advanced HNSCC. However, only a subset of HNSC patients receiving cetuximab actually responds to treatment, underlining the need for a means to tailor treatments of individual patients. The aim of the present study was to investigate the effect of cetuximab treatment on tumour growth, on tumour partial oxygen pressure as measured by LiPc electron paramagnetic resonance oximetry and on the expression of proteins involved in tumour growth, metabolism and hypoxia. Two HNSCC cell lines, UT-SCC-2 and UT-SCC-14, were used to generate xenografts on female BALB/c (nu/nu) nude mice. Mice with xenografts were given three injections of intraperitoneal cetuximab or phosphate-buffered saline, and the tumour volume was recorded continuously. After treatment the tumour partial oxygen pressure was measured by LiPc electron paramagnetic resonance oximetry and the expression of epidermal growth factor receptor (EGFR), phosphorylated EGFR, Ki-67, MCT1, MCT4, GLUT1, CAIX and HIF-1α were investigated by immunohistochemistry. In xenografts from both cell lines (UT-SCC-2 and UT-SCC-14) cetuximab had effect on the tumour volume but the effect was more pronounced on UT-SCC-14 xenografts. A higher tumour oxygenation was measured in cetuximab-treated tumours from both cell lines compared to untreated controls. Immunocytochemical staining after cetuximab treatment shows a significantly decreased expression of EGFR, pEGFR, Ki67, CAIX and nuclear HIF-1α in UT-SCC-14 tumours compared to untreated controls. MCT1 and GLUT1 were significantly decreased in tumours from both cell lines but more pronounced in UT-SCC-14 tumours. Taken together, our results show that cetuximab treatment decreases the tumour growth and increases the tumour partial oxygen pressure of HNSCC xenografts. Furthermore we found a potential connection between the partial oxygen pressure of the tumours and the expression of proteins involved in tumour growth, metabolism and hypoxia.

Protective effects of sinomenine against LPS-induced inflammation in piglets

The aim of this study was to investigate in piglets, the anti-endotoxin and anti-inflammatory effects of sinomenine, an agent commonly found in Chinese herbal medicines. In high-, middle- and low-dose sinomenine groups, piglets were initially challenged with endotoxin (i.e., 1 mg lipopolysaccharide (LPS)/kg) by intraperitoneal (IP) injection and, 3 h later, intramuscularly (IM) with sinomenine at 1, 5, or 10 mg/kg. In a drug control group, piglets were dosed IP with vehicle and 3 h late IM with 10 mg/kg sinomenine while those in an LPS control group were challenged with 1 mg LPS/kg (IP) and then vehicle 3 h later; naïve control piglets were administered normal saline IP and then IM only. At 12, 24, and 48 h post-LPS/vehicle injection, blood samples were collected from the precaval vein of piglets. Clinical signs were recorded during the trial and index levels were analyzed by ELISA kits. The results revealed sinomenine could reduce the incidence/severity of certain LPS-induced toxicities, e.g., cell adhesion, systemic inflammation, and multiple organ dysfunction. Taken together, the data suggested to us that sinomenine might effectively be useful to regulate inflammatory responses as part of future anti-endotoxin therapies.

The effect of NaCl on the level of reduced sulfur compounds in rat liver. Implications for blood pressure increase.

It is commonly known that excessive salt intake is a risk factor of hypertension. Currently, there is an increasing interest in reduced reactive sulfur species (RSS), mainly H2S and sulfane sulfur (SS) as new gasotransmitters showing vasorelaxant properties. The aim of the present study was to determine the effect of repeated administration of low sodium chloride dose included in physiological saline on blood pressure, on the level of RSS and activity of enzymes involved in their biosynthesis in the rat. Two separate experiments were carried out on male Wistar rats: one with intravenous injections of saline and the second one with intraperitoneal saline injections. Blood pressure was measured during the experiment. The level of RSS and other biochemical assays were conducted in the rat liver, because of an intense cysteine metabolism to RSS in this organ. Intravenous administration of saline induced a significant increase in systolic blood pressure while intraperitoneal injections showed only a tendency towards increasing blood pressure. The RSS (H2S and SS) level as well as the activity of the main enzyme responsible for their production in the liver of animals after iv saline injections were decreased. Animals injected with physiological saline by ip route did not reveal any statistically significant differences in SS, H2S, and activities of sulfurtransferases, although a tendency to decrease in the RSS was observed. The repeated iv saline injection induced a slight hypertension accompanied by disturbances in anaerobic cysteine metabolism in the rat liver.

Abstract 1488: Altered skeletal muscle mitochondrial function and redox biology with chemotherapy and exercise in a colorectal cancer mouse model

Abstract Background: Chemotherapy improves outcomes for patients with gastrointestinal cancer, but carries associated morbidity. Platinum-based chemotherapy adversely impacts fitness, invivo mitochondrial function, and cellular redox status in humans, however the mechanisms are poorly understood. Exercise interventions mitigate against this, but interactions between the effect of the cancer, chemotherapy, and exercise intervention are poorly understood. Using a colorectal cancer (CRC) mouse model we investigated changes in skeletal muscle oxidative phosphorylation (OxPhos) capacity and redox biology with chemotherapy and exercise. Method: SCID mice (n=48) were randomized to laparotomy with (Ca) or without (NCa) orthotopic caecal injection of DLD-1 CRC cells. CRC established over 8 weeks then mice were randomized to 1) 3 cycles of intra-peritoneal (IP) chemotherapy (Chemo; 5FU 5mg/kg, Oxaliplatin 10mg/kg and Leucovorin 90mg/kg) followed by interval exercise-training 2) Chemo and no exercise 3) IP saline sham (Sal) with exercise or 4) Sal with no exercise. Blood was collected pre and post-Chemo and at death (post-exercise), and markers of oxidative stress measured. Soleus muscle mitochondrial function was analyzed using high-resolution respirometry. Measurements included maximum OxPhos capacity (PI+II+β), Complex I and II capacities and the ratio of octanoyl-carnitine to pyruvate-supported respiration (FAO). Histopathological caecal examination was undertaken to determine cancer response. Results: Chemotherapy alone resulted in 37% lower OxPhos capacities in sham mice (p=0.011), whilst FAO was 40% lower (p=0.004). Cancer alone resulted in 38% lower OxPhos capacities (p=0.012). In combination with cancer, chemotherapy resulted in OxPhos capacities that were 42% lower than in mice with cancer but no chemotherapy (p=0.007), along with 51% lower Complex I (p&amp;lt;0.001) and 42% lower Complex II (p=0.007) capacities. In sham mice (NCaSal), exercise resulted in 33% greater Complex II activities (p=0.042), but exercise did not alter mitochondrial function in any other group. Total plasma protein content (TPPC) increased with time in the CaSal (+32%) and the NCaSal mice (+23%), whilst it decreased with chemotherapy (CaChemo -20%, NCaChemo -10%). Oxidative stress increased with chemotherapy and exercise, however exploratory interaction analyses are ongoing. Complete histopathological response was seen in 2 of 6 CaChemoEx animals. No other complete response was seen. Conclusion: Chemotherapy and cancer alone were associated with a loss of oxidative capacity, with a greater reduction when combined. Chemotherapy alone and with cancer caused a shift away from FAO and reduction in TPPC. Though we demonstrated an exercise effect in sham mice, exercise did not increase OxPhos capacity, nor did it ameliorate the effects of cancer and chemotherapy. A more intense exercise intervention may be required. Citation Format: Malcolm Andrew West, Alexandrina Roman, Ronan Astin, Sian Pugh, Bernadette Fernandez, Annette Hayden, Judith Cave, Eleanor Jaynes, John N. Primrose, Sandy Jack, Michael P. Grocott, Andrew Murray, Martin Feelisch, Alexander Mirnezami, Timothy Underwood. Altered skeletal muscle mitochondrial function and redox biology with chemotherapy and exercise in a colorectal cancer mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1488. doi:10.1158/1538-7445.AM2017-1488

POstLAparoscopic Reduction of pain By combining intraperitoneal normal salinE And the pulmonary Recruitment maneuver (POLAR BEAR trial). RCT to estimate reduction in pain after laparoscopic surgery when using a combination therapy of intraperitoneal normal saline and the pulmonary recruitment maneuver

BackgroundShoulder pain and pain in the upper abdomen are common complaints after laparoscopy, sometimes surpassing the pain at incision sites. The incidence of shoulder pain ranges from 35 to 80%. Post-laparoscopic pain is caused by retention of carbon dioxide in the abdomen, which irritates the phrenic nerve and diaphragm, causing referred pain in the shoulder and in the upper abdomen. A promising strategy to reduce this post-laparoscopic pain is the pulmonary recruitment maneuver, which indirectly increases intraperitoneal pressure and thereby facilitates removal of residual carbon dioxide. An alternative strategy is the infusion of intraperitoneal normal saline. With normal saline infusion, carbon dioxide rises and escapes through the port sites. In addition, normal saline offers a physiologic buffer system to dissolve excess carbon dioxide.Methods/DesignThis multicenter randomized controlled trial is conducted in two teaching hospitals in the Netherlands. Women between 18 and 65 years of age, with an ASA classification of I-II who are scheduled to undergo an elective laparoscopic procedure with benign gynecologic indication can participate. Following informed consent, participants are randomly allocated into two groups at the end of the surgical procedure. In the intervention group, the upper abdomen is filled with normal saline infusion with the patient in the Trendelenburg position. Then the anesthesiologist performs a standardized pulmonary recruitment maneuver with a pressure of 40 cm H2O. The trocar sleeve valves will be left open, so carbon dioxide can escape the abdominal cavity. With the patient in a neutral position the instruments are removed from the abdomen. In the control group, carbon dioxide is removed from the abdominal cavity at the end of surgery, with gentle abdominal pressure and passive exsufflation through the port sites, with open sleeve valves.The primary outcomes are the incidence and intensity of post-laparoscopic pain in the shoulder, upper abdomen and at the operation sites, at 8, 24 and 48 h after surgery. Secondary outcomes are postoperative use of analgesics, nausea, vomiting and pulmonary complications.DiscussionThis study may reduce post-laparoscopic pain in women undergoing laparoscopy.Trial registrationDutch trial register, number NTR4812.

The impact of black seed oil on tramadol-induced hepatotoxicity: Immunohistochemical and ultrastructural study

The natural herb, black seed (Nigella Sativa; NS) is one of the most important elements of folk medicine. The aim was to evaluate the impact of Nigella Sativa Oil (NSO) on the changes induced by tramadol in rat liver. Twenty four albino rats were used. Control group: given intraperitoneal and oral saline for 30days. TR-group: given intraperitoneal tramadol (20, 40, 80mg/kg/day) in the first, middle and last 10days of the experiment, respectively. TR+NS group: administered intraperitoneal tramadol in similar doses to TR-group plus oral NSO (4ml/kg/day) for 30days. Immunohistochemical, electron microscopic, biochemical and statistical studies were performed. TR-group displayed disarranged hepatic architecture, hepatic congestion, hemorrhage and necrosis. Apoptotic hepatocytes, mononuclear cellular infiltration and a significant increase in the number of anti-CD68 positive cells were observed. Ultrastructurally, hepatocytes showed shrunken nuclei, swollen mitochondria, many lysosomes and autophagic vacuoles. Activated Ito and Von Kupffer cells were also demonstrated. Elevated serum levels of AST, ALT, ALP and bilirubin were noticed. NSO administration resulted in preservation of hepatic histoarchitecture and ultrastructure and significant reductions in the number of anti-CD68 positive cells and serum levels of liver seromarkers. In conclusion, NSO administration could mitigate the alterations induced by tramadol in rat liver.

Conditional Human Immunodeficiency Virus Transactivator of Transcription Protein Expression Induces Depression-like Effects and Oxidative Stress

BackgroundThe prevalence of major depression in people with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome is substantially higher than in the general population. Mechanisms underlying this comorbidity are poorly understood. HIV transactivator of transcription (Tat) protein, produced and excreted by HIV, could be involved. We determined whether conditional Tat protein expression in mice is sufficient to induce depression-like behaviors and oxidative stress. Further, as oxidative stress is associated with depression, we determined whether decreasing or increasing oxidative stress by administering methylsulfonylmethane (MSM) or diethylmaleate, respectively, altered depression-like behavior. MethodsGT-tg bigenic mice received intraperitoneal saline or doxycycline (25–100 mg/kg/day) to induce Tat expression. G-tg mice, which do not express Tat protein, also received doxycycline. Depression-like behavior was assessed with the tail suspension test and the two-bottle saccharin/water consumption task. Reactive oxygen species and reactive nitrogen species were assessed ex vivo. Medial frontal cortex oxidative stress and temperature were measured in vivo with 9.4T proton magnetic resonance spectroscopy. ResultsTat expression increased tail suspension test immobility time in an exposure-dependent manner and reduced saccharin consumption. MSM decreased immobility time, whereas diethylmaleate increased it in saline-treated GT-tg mice. Tat and MSM behavioral effects persisted for 28 days. Tat and diethylmaleate increased reactive oxygen species and reactive nitrogen species levels, whereas MSM decreased levels. Tat expression increased medial frontal cortex glutathione levels and temperature. ConclusionsTat expression induced rapid and enduring depression-like behaviors and oxidative stress. Increasing oxidative stress increased depression-like behavior, whereas decreasing oxidative stress decreased depression-like behavior. Thus, Tat produced by HIV may contribute to high depression prevalence among people with HIV. Mitigation of oxidative stress could reduce depression severity.

Abstract 128: A Translational Strategy Targeting Type I BMP Receptors to Prevent Heterotopic Ossification

PURPOSE: Trauma-induced heterotopic ossification (tHO) is the aberrant growth of ectopic bone in soft tissue, which develops in patients following severe musculoskeletal trauma. Much of HO literature focuses on a related pathology known as fibrodysplasia ossificans progressiva (FOP), which is caused by a hyperactivating mutation in the type I bone morphogenetic protein receptor (T1-BMPR) ACVR1 (ACVR1 R206H). Consequently, emphasis has been placed on developing inhibitors with improved specificity for ACVR1. However, patients who develop tHO do not harbor known ACVR1 mutations, and it is unclear whether emphasis on ACVR1-specific inhibition is beneficial for the treatment of tHO. Here investigate whether any single T1-BMPR is required for tHO, or whether these receptors perform overlapping roles during tHO development. We further evaluate the efficacy of the BMP ligand trap, Alk3-Fc, as a broad-spectrum inhibitor of T1-BMP receptors in the treatment and prevention tHO. METHODS: Wild-type, tamoxifen-inducible Acvr1 knockout (Acvr1 tmKO: Ub.creERT/Acvr1fl/fl), tamoxifen-inducible Bmpr1a knockout (Bmpr1a tmKO: Ub.creERT/Bmpr1afl/fl), and Bmpr1b knockout (Bmpr1b-/-) mice receivedhindlimb Achille’s tendon transection and dorsal 30% total body surface area partial-thickness burn. A second cohort of wild-type mice received daily i.p. injections of Alk3-fc (2 mg/kg) or saline for 6-weeks post-injury or during weeks 0–2 (inflammation), weeks2-4 (chondrogenesis), or weeks 4–6 (ossification) after injury. Mice receiving abbreviated treatment with Alk3-fc received intraperitoneal saline injections for the remainder of their 6-week treatment period. HO was quantified by microCT 9-weeks post-injury, and cartilage formation was assessed via pentachrome staining. RESULTS: Genetic loss of Acvr1 (p>0.05), Bmpr1a (p>0.05), or Bmpr1b (p>0.05) alone was unable to significantly or substantially reduce tHO after injury. However, genetic loss of both Acvr1 and Bmpr1a (Acvr1;Bmpr1a tmKO: Ub.creERT /Acvr1fl/fl;Bmpr1afl/fl) led to a substantial and significant reduction in tHO volume (16.7-fold decrease vs. wild-type; p<0.05). Based on these findings we studied the effect of Alk3-Fc, a BMP ligand trap which inhibits BMP receptor signaling capable of broadly T1-BMPR inhibition at different points during tHO development. Mice treated daily with Alk3-Fc for 6-weeks post-injury demonstrated a significant and substantial reduction in tHO volume (2.9-fold; p<0.05). Mice treated daily with Alk3-Fc from 0–2 weeks demonstrated similar reductions in tHO volume (2.3-fold; p<0.05). This was confirmed histologically, with diminished cartilage content in 6-week and 0–2 week treated groups when compared with controls. CONCLUSIONS: Here we demonstrate that although BMP signaling is required for tHO, no single T1-BMPR (ACVR1/ALK2, BMPR1a/ALK3, or BMPR1b/ALK6) alone is necessary for this disease. This suggests that, unlike with genetic forms of HO such as FOP, these receptors have functional redundancy in the setting of tHO. Based on these data we developed an approach based on broad-spectrum T1-BMPR blockade using the ligand trap Alk3-Fc to prevent tHO formation. Our findings suggest that Alk3-Fc treatment during the earliest period after injury is sufficient to block tHO and provides a translational approach to targeting this disease process.

Evaluation of postoperative pain relief with intra-peritoneal bupivacaine instillation in laparoscopic cholecystectomy- a randomized control study

Background: Laparoscopic cholecystectomy is the surgical procedure of choice for symptomatic cholelithiasis due to the improved postoperative course, but patients undergoing laparoscopic cholecystectomy during the first 24 hours postoperatively complaints of pain. This study was designed to study the efficacy of intraperitoneal bupivacaine in reducing the initial postoperative pain and also to evaluate the postoperative shoulder tip pain, nausea and vomiting.Methods: Patients undergoing laparoscopic cholecystectomy were randomized into two groups the, study group received intraperitoneal bupivacaine 100mg at the end of the procedure and the control group received intraperitoneal saline. Post operatively patients were assessed for pulse rate, blood pressure, VAS, VRS, shoulder pain and nausea and vomiting at 0, 2, 6 and 24 hours.Results: Among the 50 patients studied, our study proved that there is no statistical significant reduction in pain with intraperitoneal bupivacaine. The VAS at 0, 2, 6 and 24 hours calculated with non-significant p-value of 0.85,0.29, 0.72 and 0.64 respectively and the VRS had p-value of 0.16, 0.08, 0.59 and 0.46 respectively and the doses of rescue analgesia consumed had p value of 0 .67, 0.61 and 0.70 which were not significant statistically.Conclusions: Instillation of 100mg bupivacaine did not significantly reduce the need for tramadol compared with saline.

Intraperitoneal adoptive transfer of mesenchymal stem cells enhances recovery from acid aspiration acute lung injury in mice

BackgroundMesenchymal stem cells (MSCs) might act as fine-tuners of inflammation during acute lung injury. We assessed the effects of adoptive transfer of MSCs in acid aspiration acute lung injury and explored the role of long pentraxin PTX3.MethodsWe conducted a prospective experimental interventional study on wild-type (WT) and PTX3-deficient (PTX3−/−) mice. Acute lung injury was induced in WT and PTX3−/− mice by instillation of hydrochloric acid into the right bronchus. One hour later, animals received intraperitoneal sterile phosphate-buffered saline (PBS), WT-MSCs (1 × 106) or PTX3−/−-MSCs (1 × 106). Twenty-four hours after injury, we measured the effects of treatments on arterial blood gases, wet/dry lung weight (W/D), CT scan analysis of lung collapse, neutrophils, TNFα and CXCL1 in bronchoalveolar lavage, and plasma PTX3. d-dimer was assayed in 1 week and OH-proline in 2 weeks to track the fibrotic evolution.ResultsIn 24 h, in comparison to PBS, WT-MSCs improved oxygenation and reduced W/D and alveolar collapse. These effects were associated with decreased concentrations of alveolar neutrophils and cytokines. WT-MSCs increased d-dimer concentration and decreased OH-proline levels, too.Treatment with PTX3−/−-MSCs ameliorated oxygenation, W/D, and alveolar TNFα, though to a lesser extent than WT-MSCs. PTX3−/−-MSCs did not improve lung collapse, neutrophil count, CXCL1, d-dimer, and OH-proline concentrations. The protective effects of WT-MSCs were dampened by lack of endogenous PTX3, too.ConclusionsIn acid aspiration acute lung injury, MSCs improve pulmonary function and limit fibrosis by fine-tuning inflammation. The role of PTX3 in determining MSCs’ effects might merit further scrutiny.