Immunomodulatory intervention in sepsis by multidrug-resistant Pseudomonas aeruginosa with thalidomide: an experimental study

BACKGROUND: Thalidomide is an inhibitor of tumour necrosis factor-alpha (TNFα) that has been proven effective for the treatment of experimental sepsis by Escherichia coli. It was tested whether it might behave as an effective immunomodulator in experimental sepsis by multidrug-resistant (MDR) Pseudomonas aeruginosa. METHODS: Sepsis was induced by the intraperitoneal injection of 1 × 10(8 )cfu/kg inoculum of the test isolate in a total of 109 Wistar rats divided in three groups as follows: group A controls; group B administered seed oil 30 minutes before bacterial challenge; and group C administered 50 mg/kg of thalidomide diluted in seed oil 30 minutes before bacterial challenge. Blood was sampled for estimation of endotoxins (LPS), TNFα, interferon-gamma (IFNγ), nitric oxide (NO) and malondialdehyde (MDA). LPS was measured by the QCL-1000 LAL assay, TNFα and IFNγ by ELISA, NO by a colorimetric assay and MDA by the thiobarbiturate assay. RESULTS: Mean (± SE) survival of groups A, B and C were 18.60 ± 1.84, 12.60 ± 0.60 and 30.50 ± 6.62 hours (p of comparisons A to C equal to 0.043 and B to C equal to 0.002). Decreased TNFα and NO levels were found in sera of animals of group C compared to group A. Plasma levels of LPS, MDA and IFNγ did not differ between groups. CONCLUSION: Intake of thalidomide considerably prolonged survival in experimental sepsis by MDR P.aeruginosa an effect probably attributed to decrease of serum TNFα

Canagliflozin attenuates the progression of atherosclerosis and inflammation process in APOE knockout mice

Background: Sodium glucose co-transporter2 inhibitors reduce the incidence of cardiovascular events in patients with type 2 diabetes mellitus based on the results of recent cardiovascular outcome studies. Herein, we investigated the efects of long-term treatment with canaglifozin on biochemical and immunohistochemical markers related to atherosclerosis and atherosclerosis development in the aorta of apolipoprotein E knockout (Apo-E(−/−) ) mice. Methods: At the age of 5 weeks, mice were switched from normal to a high-fat diet. After 5 weeks, Apo-E(−/−) mice were divided into control-group (6 mice) treated with 0.5% hydroxypropyl methylcellulose and Cana-group (7 mice) treated with canaglifozin (10 mg/kg per day) per os. After 5 weeks of intervention, animals were sacrifced, and heart and aorta were removed. Sections stained with hematoxylin–eosin (H&E) were used for histomorphometry whereas Masson’s stained tissues were used to quantify the collagen content. Immunohistochemistry to assess MCP-1, CD68, a-smooth muscle actin, MMP-2, MMP-9, TIMP-1 and TIMP-2 expression was carried out and q-PCR experiments were performed to quantify mRNA expression. Results: Canaglifozin-group mice had lower total-cholesterol, triglycerides and glucose levels (P<0.01), while heart rate was signifcantly lower (P<0.05). Histomorphometry revealed that one in seven Cana-group mice versus four in six control mice developed atheromatosis, while aortic root plaque was signifcantly less, and collagen was 1.6 times more intense in canaglifozin-group suggesting increased plaque stability. Immunohistochemistry revealed that MCP-1 was signifcantly less expressed (P<0.05) in the aortic root of canaglifozin-group while reduced expression of a-actin and CD68 was not reaching signifcance (P=0.15). VCAM-1 and MCP-1 mRNA levels were lower (P=0.02 and P=0.07, respectively), while TIMP-1/MMP-2 ratio expression was higher in canaglifozin-group approaching statistical signifcance (P=0.07). Conclusions: Canaglifozin attenuates the progression of atherosclerosis, reducing (1) hyperlipidemia and hyper‑ glycemia, and (2) infammatory process, by lowering the expression of infammatory molecules such as MCP-1 and VCAM-1. Moreover, canaglifozin was found to increase the atherosclerotic plaque stability via increasing TIMP-1/ MMP-2 ratio expression

Liver regeneration: Focus on cell types and topographic differences

Background/Aims: The liver has a remarkable capacity to regenerate after injury or resection. The aim of the current review is to outline the morphological changes at the cellular level and the intralobular differences during the regenerative response after partial hepatectomy. Methods: Relevant studies were reviewed using the Medline database. Results: At 24 h after partial hepatectomy, many alterations between the periportal and pericentral regions of the hepatic lobule occur, which coincide with the peak of hepatocellular proliferation in the periportal areas. Questions unanswered involve the precise role of Kupffer cells and whether the regenerative process is characterized by a net increase in the number of lobules or by an increase in the size of the existing lobules. Conclusions: Liver regeneration has been studied for many years, but further research is essential, since in-depth knowledge of the mechanisms that govern the regenerative process may expand the treatment options in patients with hepatic disease. © 2009 S. Karger AG, Basel

A biomechanical study of the role of sitagliptin on the bone characteristics of diabetic rats

An experimental protocol is described aiming to explore the influence of Type 2 Diabetes Mellitus on the biomechanical response of the bone tissue and, also, to quantify the potential beneficial role of a pharmaceutical treatment, based on sitagliptin, a diabetes drug that increases the levels of natural substances called incretins. Twenty eight male, 10-week old Wistar rats were used, divided into three groups, i.e., the control one, the group including the diabetic rats and, finally, the group including the diabetic rats which were treated using sitagliptin. The biomechanical study was based on a series of three-point bending tests of the femora of the sacrificed rats and the analysis of the experimental data was implemented in terms of the actual geometry of the fractured cross-section. It was concluded that diabetic bones undertake larger forces despite the fact that the “diameter” of their crosssection was somehow smaller. On the contrary, the slope of the load-deflection curve (corresponding to a measure of the stiffness) of diabetic bones is slightly lower compared to the control bones. Finally, it seems that treating diabetic animals with sitagliptin only partly reverses the effect of Type 2 Diabetes Mellitus on their bone tissue, at least concerning its strength and stiffness. © 2020, Gruppo Italiano Frattura. All rights reserved

A combined cellular and surgical ventricular reconstruction therapeutic approach produces attenuation of remodeling in infarcted rats

Background: Left ventricular reconstruction (LVR) has been shown to provide transient benefits to the LV structure and function of infarcted hearts; however, long-term results have been disappointing as LVR-induced benefits are typically not sustained. We hypothesized that administration of cardiosphere-derived cells (CDCs), which promote myocardial repair and regeneration, may result in long-term preservation of the beneficial effects of LVR in ischemic cardiomyopathy. Methods: Wistar Kyoto rats underwent myocardial infarction (MI) and two weeks later were randomized into 3 groups: in Group 1 (n=9), LVR was performed by plication of the infarcted apex and CDCs were injected in the infarct border zone (IBZ); group 2 animals (n=9) underwent LVR and received vehicle solution in the IBZ; and Group 3 animals (n=10) were injected with vehicle solution in the IBZ without undergoing LVR. Echocardiograms were performed at baseline, 4 days post-apex plication, and at 3 months post-MI. Results: At baseline, all animal groups had a comparable LVEF, LV end-diastolic volume (EDV) and LV end-systolic volume (ESV). Four days post-LV apex plication, Group 1 and Group 2 animals exhibited comparable significant improvement in EF and comparable significant reduction in LVEDV and LVESV. Three months post-MI, Group 1 animals had a decreased LVEDV, decreased LVESV, less impaired CS, increased peak systolic torsion and increased EF compared to animals in Groups 2 and 3. Conclusion: In infarcted rat hearts, intramyocardial delivery of CDCs in conjunction with LVR resulted in significant and sustained amelioration of LV remodeling and improvement in LV function compared to LVR alone. (C) 2016 Hellenic Society of Cardiology. Publishing services by Elsevier B.V

Effective Immunomodulatory Treatment of Escherichia coli Experimental Sepsis with Thalidomide

Thalidomide, an agent which inhibits biosynthesis of tumor necrosis factor alpha (TNF-α) and which is used to treat a variety of chronic inflammatory conditions, was investigated as therapy for experimental sepsis. Sepsis was induced by intraperitoneal injection of 10(7) CFU of Escherichia coli per kg of body weight to 80 Wistar rats divided into four groups. Group A consisted of 24 control animals that did not receive any pretreatment, group B consisted of 18 vehicle-treated control animals pretreated with seed oil, group C consisted of 30 rats administered thalidomide diluted in seed oil at a dose of 50 mg/kg 30 min before bacterial challenge, and group D consisted of eight animals that were not challenged with E. coli but that were used for white blood cell count determination. Sepsis was determined by measurement of vital signs before and 6 h after bacterial challenge. After 6 h the animals were killed and blood was sampled for culture; white blood cell count determination; and determination of endotoxin (lipopolysaccharide), TNF-α, interleukin-1β (IL-1β), and IL-6 levels. The levels of these cytokines were also estimated in the supernatants of human monocytes pretreated with thalidomide after exposure to the isolate. Sepsis developed in all vehicle-treated control animals and controls by 6 h after bacterial challenge but in only 10 animals (33.3%) pretreated with thalidomide (P < 0.0001). Six hours after bacterial challenge all animals had similar levels of endotoxemia, IL-1β, and IL-6. The mean white blood cell count for groups A, B, and C were 5,631.1, 2,638.9, and 8,169.3 cells/μl, respectively (P value between groups, <0.0001); the TNF-α levels were 77.3, 107.2, and 26.1 pg/ml, respectively (P values between groups, <0.0001). Pretreatment of human monocytes with thalidomide prevented the secretion of TNF-α and IL-1β but not that of IL-6. It is concluded that thalidomide exerts a considerable anti-inflammatory effect by preventing evolution to sepsis and by decreasing the level of production of TNF-α and therefore deserves to be further evaluated in research for the therapy of sepsis