Non-Invasive Targeted Peripheral Nerve Ablation Using 3D MR Neurography and MRI-Guided High-Intensity Focused Ultrasound (MR-HIFU): Pilot Study in a Swine Model

Purpose

Ultrasound (US)-guided high intensity focused ultrasound (HIFU) has been proposed for noninvasive treatment of neuropathic pain and has been investigated in in-vivo studies. However, ultrasound has important limitations regarding treatment guidance and temperature monitoring. Magnetic resonance (MR)-imaging guidance may overcome these limitations and MR-guided HIFU (MR-HIFU) has been used successfully for other clinical indications. The primary purpose of this study was to evaluate the feasibility of utilizing 3D MR neurography to identify and guide ablation of peripheral nerves using a clinical MR-HIFU system.

Methods

Volumetric MR-HIFU was used to induce lesions in the peripheral nerves of the lower limbs in three pigs. Diffusion-prep MR neurography and T1-weighted images were utilized to identify the target, plan treatment and immediate post-treatment evaluation. For each treatment, one 8 or 12 mm diameter treatment cell was used (sonication duration 20 s and 36 s, power 160–300 W). Peripheral nerves were extracted < 3 hours after treatment. Ablation dimensions were calculated from thermal maps, post-contrast MRI and macroscopy. Histological analysis included standard H&E staining, Masson’s trichrome and toluidine blue staining.

Results

All targeted peripheral nerves were identifiable on MR neurography and T1-weighted images and could be accurately ablated with a single exposure of focused ultrasound, with peak temperatures of 60.3 to 85.7°C. The lesion dimensions as measured on MR neurography were similar to the lesion dimensions as measured on CE-T1, thermal dose maps, and macroscopy. Histology indicated major hyperacute peripheral nerve damage, mostly confined to the location targeted for ablation.

Conclusion

Our preliminary results indicate that targeted peripheral nerve ablation is feasible with MR-HIFU. Diffusion-prep 3D MR neurography has potential for guiding therapy procedures where either nerve targeting or avoidance is desired, and may also have potential for post-treatment verification of thermal lesions without contrast injection.     

Enzymes from spent mushroom substrate of <Emphasis Type="Italic">Pleurotus sajor</Emphasis>-<Emphasis Type="Italic">caju</Emphasis> for the decolourisation and detoxification of textile dyes

The potential of ligninolytic enzymes, including lignin peroxidase (LiP) as the main enzyme from the spent mushroom substrate of Pleurotus sajor-caju was evaluated for the decolourisation of five dyes from azo and anthraquinone dye groups. Among the azo dyes, reactive black 5 and reactive orange 16 were 84.0 and 80.9% decolourised respectively, after 4 h of incubation with 45 U of LiP as compared to 32.1% decolourisation of disperse blue 79. Among the anthraquinone dyes, disperse red 60 was decolourised to 47.2% after 4 h of incubation with 45 U of LiP as compared to 5.9% decolourisation of disperse blue 56. Increasing the LiP concentration and incubation time had a positive effect on the decolourisation of anthraquinone dyes as compared to azo dyes. A 67.9% decolourisation of synthetic textile waste-water was achieved after 4 h of incubation with 25 U of LiP. Increasing the incubation time significantly increased (P < 0.05) the decolourisation of synthetic textile waste-water. Further, there was a 52.4% reduction in the toxicity of synthetic textile waste-water treated with 55 U of LiP for 4 h. However, only 35.7% reduction in toxicity was achieved when the synthetic textile waste-water was treated with 55 U of LiP for 24 h. In this study, it was shown that the spent mushroom substrate of P. sajor-caju could be a cheap source of ligninolytic enzymes for the decolourisation of dyes in textile industry wastewaters.     

Immunoregulatory role of CD1d in the hydrocarbon oil-induced model of lupus nephritis

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease that is accompanied by the emergence of autoreactive T cells and a reduction in regulatory T cells. Humans and mice with SLE have reduced numbers of CD1d-restricted NK T cells, suggesting a role for these cells in the regulation of SLE. In this study, we show that CD1d deficiency exacerbates lupus nephritis induced by the hydrocarbon oil pristane. This exacerbation in disease is associated with: 1) reduced TNF-alpha and IL-4 production by T cells, especially during the disease induction phase; and 2) expansion of marginal zone B cells. Strikingly, inoculation of pristane in wild-type mice resulted in reduced numbers and/or functions of NK T cells and CD1d-expressing dendritic cells. These findings suggest that CD1d may play an immunoregulatory role in the development of lupus in the pristane-induced model.     

Quantitative and qualitative differences in the in vivo response of NKT cells to distinct alpha- and beta-anomeric glycolipids

NKT cells represent a unique subset of immunoregulatory T cells that recognize glycolipid Ags presented by the MHC class I-like molecule CD1d. Because of their immunoregulatory properties, NKT cells are attractive targets for the development of immunotherapies. The prototypical NKT cell ligand alpha-galactosylceramide (alpha-GalCer), originally isolated from a marine sponge, has potent immunomodulatory activities in mice, demonstrating therapeutic efficacy against metastatic tumors, infections, and autoimmune diseases, but also has a number of adverse side effects. In vivo administration of alpha-GalCer to mice results in the rapid activation of NKT cells, which is characterized by cytokine secretion, surface receptor down-regulation, expansion, and secondary activation of a variety of innate and adaptive immune system cells. In this study, we have evaluated the in vivo immune response of mice to a set of structural analogues of alpha-GalCer. Our results show that, contrary to current thinking, beta-anomeric GalCer can induce CD1d-dependent biological activities in mice, albeit at lower potency than alpha-anomeric GalCer. In addition, we show that the response of NKT cells to distinct GalCer differs not only quantitatively, but also qualitatively. These findings indicate that NKT cells can fine-tune their immune responses to distinct glycolipid Ags in vivo, a property that may be exploited for the development of effective and safe NKT cell-based immunotherapies.     

Probiotic Potential of a Lactobacillus Bacterium of Canine Faecal-Origin and Its Impact on Select Gut Health Indices and Immune Response of Dogs

The objective of the present study was to develop a probiotic of canine-origin for its potential application in pet nutrition. Accordingly, 32 lactic acid bacteria (LAB) strains were isolated from faeces of dogs, out of which 9 strains were short-listed for further in vitro testing based on the aggregation time and cell surface hydrophobicity. The results of acid-, bile- and phenol-tolerance tests indicated that out of the nine, isolate cPRO23 was having better resistance to these adverse conditions likely to be encountered in the gastrointestinal tract. The isolate also showed optimal enzymatic activities for amylase, lipase and protease. Further assessments also indicated its superiority in terms of co-aggregation and antagonistic activity against pathogenic strains of Salmonella typhimurium and Salmonella enteritidis. Subsequently, the isolate was identified through 16S rRNA sequencing and sequence homology, and designated as Lactobacillus johnsonii CPN23. The candidate probiotic was then evaluated in vivo using 15 adult Labrador dogs, divided into 3 groups, viz. CON (with no probiotics), dPRO (with Lactobacillus acidophilus NCDC 15 as a conventional dairy-origin probiotic) and cPRO (with L. johnsonii CPN23 as a canine-origin probiotic). Results of the 9-week study indicated that supplementation of cPRO improved (P < 0.05) the faecal concentration of acetate and butyrate with a concomitant reduction (P < 0.05) in faecal ammonia. The cell-mediated immune response, assessed as delayed-type hypersensitivity reaction to phytohaemagglutinin-P, was better (P < 0.05) in dogs fed cPRO as compared to the CON dogs. There were, however, no variations evident in the antibody response to sheep-erythrocytes among the three groups. It is concluded that the canine-origin L. johnsonii CPN23, in addition to possessing all the in vitro functional attributes of a candidate probiotic, also has the potential to be used as a probiotic in pet nutrition programs.     

Immunotherapy with ligands of natural killer T cells

Natural killer T (NKT) cells are innate lymphocytes that share receptor structures and functions with conventional T cells and natural killer cells. NKT cells are specific for glycolipid antigens bound by the major histocompatibility complex class I-like protein CD1d. One striking property of NKT cells is their capacity to rapidly produce large amounts of cytokines in response to T-cell receptor engagement, suggesting that activated NKT cells can modulate adaptive immune responses. Recent pre-clinical studies have revealed significant efficacy of NKT-cell ligands such as the glycolipid alpha-galactosylceramide for treatment of metastatic cancers and infections, and for prevention of autoimmune diseases. These findings suggest that appropriate stimulation of NKT cells could be exploited for prevention or treatment of human diseases.     

Characterization and expression analysis of chalcone synthase and chalcone isomerase genes in Phyllanthus emblica (L.)

Journal of Plant Biochemistry and Biotechnology - The two decisive enzymes in flavonoid biosynthetic pathway are chalcone synthase (CHS) and chalcone isomerase (CHI), wherein the former carries the...