A Comparative Study of the Effect of Clonidine Tramadol and Nalbuphine on Postspinal Anaesthesia Shivering

OBJECTIVES: To compare the efficacy of Tramadol, Nalbuphine, and Clonidine in the treatment of shivering after spinal anaesthesia in caesarean section. METHODS: A prospective randomised comparative study was conducted between 1.8.2013 to 1.9.2014 to compare the efficacy of Tramadol, Nalbuphine, Clonidine in treatment of shivering after spinal anaesthesia in casearean section. Patients included in this study are those who developed shivering after spinal anaesthesia for caesarean section. Patients were randomly allotted to one of the three groups, namely T group (25) who received Tramadol 0.5mg/kg i.v., N group (25) who received Nalbuphine 0.1mg/kg i.v and C group (25)who received clonidine 0.5μg/kg iv. Vital parameters of the patient such as H.R. B.P. SPO2, RR and temperature were monitored at regular intervals as per protocol. Events such as onset of shivering, time taken to stop shivering, recurrence of shivering and side effects like nausea, vomiting bradycardia, hypotension and sedation were also noted. Statistical tests like chi square test, Anova test were applied to the data collected. RESULTS: Among the 75 patients who developed shivering of grades 3 & 4 requiring treatment were randomly allotted to one of the three group. The mean temperature at which patient developed shivering was 36.4°C, and the mean duration of shivering to occur following spinal anaesthesia was 22.5 mts. Tramadol 0.5 mg/kg controlled shivering in mean time of 4 minutes, clonidine 0.5 μg/kg, controlled shivering in mean time of 2 minutes and Nalbuphine 0.1 mg/kg controlled shivering in mean time of 4 minutes. CONCLUSION: Our study concludes that all three drugs namely Tramadol clonidine and nalbuphine were effective in controlling postspinal anesthesia shivering. Among them clonidine took lesser time to achieve complete cessation of shivering and also maintained better hemodynamics throughout the study

ILB® resolves inflammatory scarring and promotes functional tissue repair

Fibrotic disease is a major cause of mortality worldwide, with fibrosis arising from prolonged inflammation and aberrant extracellular matrix dynamics. Compromised cellular and tissue repair processes following injury, infection, metabolic dysfunction, autoimmune conditions and vascular diseases leave tissues susceptible to unresolved inflammation, fibrogenesis, loss of function and scarring. There has been limited clinical success with therapies for inflammatory and fibrotic diseases such that there remains a large unmet therapeutic need to restore normal tissue homoeostasis without detrimental side effects. We investigated the effects of a newly formulated low molecular weight dextran sulfate (LMW-DS), termed ILB®, to resolve inflammation and activate matrix remodelling in rodent and human disease models. We demonstrated modulation of the expression of multiple pro-inflammatory cytokines and chemokines in vitro together with scar resolution and improved matrix remodelling in vivo. Of particular relevance, we demonstrated that ILB® acts, in part, by downregulating transforming growth factor (TGF)β signalling genes and by altering gene expression relating to extracellular matrix dynamics, leading to tissue remodelling, reduced fibrosis and functional tissue regeneration. These observations indicate the potential of ILB® to alleviate fibrotic diseases

3D human liver tissue from pluripotent stem cells displays stable phenotype in vitro and supports compromised liver function in vivo.

Liver disease is an escalating global health issue. While liver transplantation is an effective mode of therapy, patient mortality has increased due to the shortage of donor organs. Developing renewable sources of human liver tissue is therefore attractive. Pluripotent stem cell-derived liver tissue represents a potential alternative to cadaver derived hepatocytes and whole organ transplant. At present, two-dimensional differentiation procedures deliver tissue lacking certain functions and long-term stability. Efforts to overcome these limiting factors have led to the building of three-dimensional (3D) cellular aggregates. Although enabling for the field, their widespread application is limited due to their reliance on variable biological components. Our studies focused on the development of 3D liver tissue under defined conditions. In vitro generated 3D tissues exhibited stable phenotype for over 1 year in culture, providing an attractive resource for long-term in vitro studies. Moreover, 3D derived tissue provided critical liver support in two animal models, including immunocompetent recipients. Therefore, we believe that our study provides stable human tissue to better model liver biology 'in the dish', and in the future may permit the support of compromised liver function in humans

Isoaspartate, Carbamoyl phosphate synthase-1, and carbonic anhydrase-III as biomarkers of liver injury

We had previously shown that alcohol consumption can induce cellular isoaspartate protein damage via an impairment of the activity of protein isoaspartyl methyltransferase (PIMT), an enzyme that triggers repair of isoaspartate protein damage. To further investigate the mechanism of isoaspartate accumulation, hepatocytes cultured from control or 4-week ethanol-fed rats were incubated in vitro with tubercidin or adenosine. Both these agents, known to elevate intracellular S-adenosylhomocysteine levels, increased cellular isoaspartate damage over that recorded following ethanol consumption in vivo. Increased isoaspartate damage was attenuated by treatment with betaine. To characterize isoaspartate-damaged proteins that accumulate after ethanol administration, rat liver cytosolic proteins were methylated using exogenous PIMT and 3H-S- adenosylmethionine and proteins resolved by gel electrophoresis. Three major protein bands of ~75-80 kDa, ~95-100 kDa, and ~155-160 kDa were identified by autoradiography. Column chromatography used to enrich isoaspartate-damaged proteins indicated that damaged proteins from ethanol-fed rats were similar to those that accrued in the livers of PIMT knockout (KO) mice. Carbamoyl phosphate synthase-1 (CPS-1) was partially purified and identified as the ~160kDa protein target of PIMT in ethanol-fed rats and in PIMT KO mice. Analysis of the liver proteome of 4-week ethanol-fed rats and PIMT KO mice demonstrated elevated cytosolic CPS-1 and betaine homocysteine S-methyltransferase-1 when compared to their respective controls, and a significant reduction of carbonic anhydrase-III (CA-III) evident only in ethanol-fed rats. Ethanol feeding of rats for 8 weeks resulted in a larger (~2.3-fold) increase in CPS-1 levels compared to 4- week ethanol feeding indicating that CPS-1 accumulation correlated with the duration of ethanol consumption. Collectively, our results suggest that elevated isoaspartate and CPS-1, and reduced CA-III levels could serve as biomarkers of hepatocellular injury

Administration of Human MSC-Derived Extracellular Vesicles for the Treatment of Primary Sclerosing Cholangitis: Preclinical Data in MDR2 Knockout Mice.

Primary Sclerosing Cholangitis (PSC) is a progressive liver disease for which there is no effective medical therapy. PSC belongs to the family of immune-mediated biliary disorders and it is characterized by persistent biliary inflammation and fibrosis. Here, we explored the possibility of using extracellular vesicles (EVs) derived from human, bone marrow mesenchymal stromal cells (MSCs) to target liver inflammation and reduce fibrosis in a mouse model of PSC. Five-week-old male FVB.129P2-Abcb mice were intraperitoneally injected with either 100 µL of EVs (± 9.1 × 10 particles/mL) or PBS, once a week, for three consecutive weeks. One week after the last injection, mice were sacrificed and liver and blood collected for flow cytometry analysis and transaminase quantification. In FVB.129P2-Abcb4 mice, EV administration resulted in reduced serum levels of alkaline phosphatase (ALP), bile acid (BA), and alanine aminotransferase (ALT), as well as in decreased liver fibrosis. Mechanistically, we observed that EVs reduce liver accumulation of both granulocytes and T cells and dampen VCAM-1 expression. Further analysis revealed that the therapeutic effect of EVs is accompanied by the inhibition of NFkB activation in proximity of the portal triad. Our pre-clinical experiments suggest that EVs isolated from MSCs may represent an effective therapeutic strategy to treat patients suffering from PSC

Chimeric apolipoproteins mediated cholesterol efflux from J774 macrophages.

<p><b>Panels A and B.</b> J774 macrophages were labeled with [³H]cholesterol, and treated in the absence (open bars) or presence (closed bars) of cAMP. Lipid-free chimeras or parent apolipoproteins (10 μg/ml) were added to cells in serum-free RPMI-1640 medium, and amount of cholesterol efflux determined at 4 h. Conditions of time and dose were within a linear response range to allow for potential differences (if any) to be observed with and without treatment of acceptors in the absence (<b>Panel A</b>) or presence (<b>Panel B</b>) of 5x molar excess of BME. <b>Panel C.</b> Dependence of cholesterol efflux on chimera concentration. The cells were treated with the indicated concentrations of the chimeras for 4 h. <b>Panel D.</b> Kinetics of chimera-mediated cholesterol efflux. Chimera-mediated cholesterol efflux was followed at the indicated time points using 32 μg/μl of each protein. Values are means ±SD from triplicate determinations within a single experiment representative of two. For <b>Panel C</b> and <b>Panel D</b>, apoE3-NT/apoAI-CT (open inverted triangles); apoAI-NT/apoE-CT (open circles).</p

Characterization of chimeric apolipoproteins.

<p><b>Panel A</b>. SDS-PAGE analysis of the chimeric apolipoproteins. Electrophoresis of chimeric and parent proteins (20 μg protein) was carried out using a 4–20% acrylamide gradient Tris-glycine gel under reducing conditions in the presence of BME. <b>Panel B</b>. Western blot analysis of chimeras (0.5 μg protein) using mouse HRP-conjugated apoE polyclonal antibody (<i>Left</i>) or apoAI antibody (<i>Right</i>). Lane assignments are as follows: Lane 1, apoAI; Lanes 2, apoAI-NT/apoE-CT; Lanes 3, apoE3; Lanes 4, apoE3-NT/apoAI-CT.</p