Plasmodium-Salmonella Coinfection Induces Intense Inflammatory Response, Oxidative Stress, and Liver Damage: A Mice Model Study for Therapeutic Strategy.

Impairment of host immune response in malaria favors bacteremia caused by typhoidal or nontyphoidal serovars of Salmonella enterica. Ofloxacin and Artesunate are the drugs that are clinically proven for treating typhoid and malaria, respectively. The study evaluates the host responses upon treatment with antibiotic (Ofloxacin) and antimalarial (Artesunate) in a standardized mice model harboring coinfection. BALB/c mice (18-22 g) were simultaneously coinfected with Plasmodium yoelii nigeriensis (Pyn) and S. enterica serovar Typhimurium (STm) and then treated with Ofloxacin or/and Artesunate from day 4 to day 7. The bacterial burden, liver function enzymes, oxidative stress, m-RNA expression of Toll-like receptors (TLR-2 and TLR-4), Th1/Th2 cytokines, hemeoxygenase-1, and NFкB were assessed. Ofloxacin treatment failed to counter the bacterial proliferation in Pyn-STm coinfected mice. However, upon controlling parasitemia with antimalarial, the efficacy of Ofloxacin could be regained. Elevated bacterial burden with malaria induces the expression of TLR-2 and TLR-4 triggering intense inflammatory response (NFκB, Th1/Th2 cytokines) in coinfected mice. This results in critical liver damage (ALT, AST, and ALP), oxidative stress (lipid peroxidation, total GSH, catalase, and super oxide dismutase), and hemeoxygenase-1 (HO-1). The study concludes that malaria infection aggravates the secondary infection of Salmonella serovars and the control of septicemia is critical in recovery of the coinfected subject.

Potential for adaptive dose escalation in radiotherapy for patients with locally advanced non-small-cell lung cancer in a low mid income setting.

OBJECTIVE: To evaluate the effect of tumour volume regression on adaptive treatment planning, reduction in doses to organs at risk (OARs) and dose escalation. METHODS: 20 patients undergoing radical chemoradiotherapy were imaged in the fifth week of radiotherapy (CT_45) to evaluate differences in tumour volume regression between concurrent and sequential chemoradiotherapy. Replanning was carried out in the CT_45 in those with >20% regression (n = 10) and evaluated for change in target coverage indices (the coverage index and external volume index) and doses to the OAR [mean lung dose, V20 and V5 of whole and ipsilateral lung (MLDWL, V20WL, V5WL, MLDIL, V20IL, V5IL); mean oesophagus dose, V50oesophagus; and maximum spinal cord doses]. The feasibility of maximum dose escalation was explored keeping the limit of the OAR below their tolerance limits. RESULTS: Tumour regression was higher with concurrent chemoradiotherapy as compared with sequential chemoradiotherapy (p = 0.02). With the adaptive plan, the mean coverage index improved from 0.96 (±0.14) to 1.29 (±0.36), the mean external volume index changed from 1.39(±0.60) to 1.41(±0.56) and the reduction in doses to the OARs were MLDWL 10.6%, V20WL 1.3%, V5WL 1.2%, MLDIL 6.6%, V20IL 1.5%, V5IL 2.3%, mean oesophagus dose 7%, V50oesophagus 31% and maximum cord dose 0.35%. Dose escalation was possible in four patients in CT_45. CONCLUSION: There is 35% reduction in tumour volume with chemoradiotherapy at 45 Gy which allows improvement in conformality, reduction in doses to the OARs and dose escalation in 40% of patients. Advances in knowledge: This article emphasizes that adaptive planning with a single diagnostic scan at 45 Gy has the potential for improvement of radiotherapy planning indices, dose escalation while respecting the dose to the OAR. This simple strategy can be helpful in radiotherapy planning upto 60 Gy in 40% of the patients of locally advanced non-small-cell lung cancer in countries with limited resources.

Diarylheptanoids from Alnus nepalensis attenuates LPS-induced inflammation in macrophages and endotoxic shock in mice.

Diarylheptanoids, a group of plant secondary metabolites are increasingly recognized as potential therapeutic agents. The aim of study was to ascertain the anti-inflammatory profile of diarylheptanoids from Alnus nepalensis against lipopolysaccharide (LPS)-induced inflammation in macrophages and endotoxic shock in mice. Extracts prepared from dried leaves of A. nepalensis using standard solvents were tested against LPS-induced inflammation in macrophages. Among all, butanol extract (ANB) has shown most significant inhibition of pro-inflammatory cytokines without any cytotoxicity. HPLC analysis of ANB showed the presence of diarylheptanoids. The diarylheptanoids were further isolated and tested in-vitro for anti-inflammatory activity. Treatment of isolated diarylheptanoids (HOG, ORE and PLS) was able to reduce the production and mRNA level of pro-inflammatory cytokines (TNF-α and IL-6). Furthermore, we demonstrated that it inhibited the expression of NF-kB protein in LPS-induced inflammation in macrophages. In-vivo efficacy and safety profile of ANB revealed that oral treatment of ANB was able to improve the survival rate, and inhibited the production of pro-inflammatory cytokines in serum, attenuated vital organ injury in a dose dependent manner without any toxic effect at higher dose in mice. The results suggest that diarylheptanoids from A. nepalensis can be considered as potential therapeutic candidates for the management of inflammation related diseases.

Flavonoids rich fraction of Citrus limetta fruit peels reduces proinflammatory cytokine production and attenuates malaria pathogenesis.

Exploration of possible pharmacological effects along with characterisation of the bioactive compounds present in peels may have a key role in converting the fruit waste materials into therapeutic value added products. Extracts prepared from the Citrus limetta fruit peels were studied for antioxidant and anti- inflammatory activity using in-vitro bioassays. Among all, ClEt an ethanol extract of Citrus limetta fruit peels has shown promising anti-oxidant and anti-inflammatory activity. ClEt was further validated to ensure its safety evaluation at 2000mg/kg and anti-malarial efficacy at 100, 250, 500 mg/kg body weight with special reference to inflammatory mediators involved in malaria pathogenesis. In-vivo study revealed that ClEt was safe at higher dose and showed promising anti-malarial activity by inhibiting the parasitaemia and inflammatory mediators (IFN-γ, TNF-α, IL-6) involved in malaria pathogenesis, able to improve the haemoglobin and glucose level and increase the survival time. Chemical fingerprint of ClEt revealed the presence of flavonoids. Results suggested the suitability of ClEt, a flavonoid rich fraction of Citrus limetta fruit peels as a candidate for further investigation towards the management of malaria pathogenesis.

α-(-)-bisabolol reduces pro-inflammatory cytokine production and ameliorates skin inflammation.

α-(-)-bisabolol is a natural monocyclic sesquiterpene present in the essential oil has generated considerable interest in the chemical and pharmaceutical industries and currently in use in various formulations, mainly in cosmetics. This study was undertaken to evaluate its therapeutic profile against skin inflammation using in-vitro, in-vivo and in-silico assays. Lipopolysachharide (LPS) and 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced production of proinflammatory cytokines (TNF-α and IL-6) in macrophage cells as well as in TPA-induced skin inflammation in mice was significantly inhibited by α-(-)-bisabolol. TPA-induced ear thickness, ear weight and lipid peroxidation and histopathological damage in the ear tissue were also significantly inhibited by topical application of α-(-)-bisabolol in a dose dependent manner. In-vitro and in-vivo toxicity profiles indicate that it is safe for topical application on skin. Molecular docking study also revealed its strong binding affinity to the active site of the pro-inflammatory proteins. These findings suggested that α-(-)-bisabolol may be a useful therapeutic candidate for the treatment of skin inflammation.

Synthesis of diosgenin analogues as potential anti-inflammatory agents.

We herein report the synthesis of diosgenin analogues from commercially available diosgenin as the starting material. The structures of newly synthesised compounds were confirmed by (1)H NMR, (13)C NMR and mass spectrometry. All analogues were evaluated for in-vitro anti-inflammatory profile against LPS-induced inflammation in primary peritoneal macrophages isolated from mice by quantification of pro-inflammatory (TNF-α, IL-6 and IL-1ß) cytokines in cell culture supernatant using the ELISA technique followed by in-vitro cytotoxicity study. Among the synthesised analogues, analogue 15 [(E) 26-(3',4',5'-trimethoxybenzylidene)-furost-5en-3ß-acetate)] showed significant anti-inflammatory activity by inhibiting LPS-induced pro-inflammatory cytokines in a dose-dependent manner without any cytotoxicity. Efficacy and safety of analogue 15 were further validated in an in-vivo system using LPS-induced sepsis model and acute oral toxicity in mice. Oral administration of analogue 15 inhibited the pro-inflammatory cytokines in serum, attenuated the liver and lung injury and reduced the mortality rate in sepsis mice. Acute oral toxicity study showed that analogue 15 is non-toxic at higher dose in BALB/c mice. Molecular docking study revealed the strong binding affinity of diosgenin analogues to the active site of the pro-inflammatory proteins. These findings suggested that analogue 15 may be a useful therapeutic candidate for the treatment of inflammatory diseases.

Protective mechanism of lignans from Phyllanthus amarus against galactosamine/ lipopolysaccharide-induced hepatitis: an in-vivo and in-silico studies.

Phyllanthus amarus is a medicinal herb used in traditional Indian medicine for liver disorders. Several researches also show that it acts primarily in the liver, but the molecules were unidentified for liver protective activity. This study was to determine whether the lignans isolated from P. amarus attenuates the D-galactosamine (GalN) / Lipopolysaccharide (LPS)- induced acute hepatitis in mice. Standardize mixture of lignans (slPA) isolated from leaves of P. amarus using automated chromatographic technique was used for experiments. Experimental mice were orally pre-treated with slPA (10, 30 and 100 mg/kg) for 7 days before intra-peritoneal injection of GalN/LPS. Acute hepatitis in mice was confirmed by significant increase of pro-inflammatory cytokines, and hepatotoxic markers. Pre-treatment of slPA exhibit significant liver protection in dose dependant mannaer. In-silico molecular docking studies also suggests that lignans are preferentially more active due to strong binding affinity against pro-inflammatory cytokines; IL-1ß, IL-6, and TNF-α. The electronic parameters of lignans for bioavailability, drug likeness and toxicity were within the acceptable limit. In-vivo and in-silico results suggest that pretreatment of slPA exhibit potent hepatoprotection against GalN/LPS-induced hepatitis in mice and the liver protective effects may be due to the inhibition of inflammatory mediators.

Molecular docking and ADME studies of natural compounds of Agarwood oil for topical anti-inflammatory activity.

Aquilaria agallocha Roxb. family, Thymelaeaceae, is an evergreen plant of South-East Asia, commonly described as aloe wood or agarwood. Traditionally, the bark, root and heartwood are used for their medicinal properties as a folk medicine for hundreds of years. Chemical analyses revealed that the bulk of the oil is constituted by agarospirol (12.5%), jinkoh-eremol (11.8%) and hinesol (8.9%) as major contributor. In the present work, a QSAR model for antiinflammatory activity of 10-epi-γ-Eudesmol, jinkoh-eremol, agarospirol and other compounds has been developed by multiple linear regression method. The r(2) and rCV(2) of a model were 0.89 and 0.81 respectively. In silico molecular docking study suggests that compound 10-epi-γ-Eudesmol, jinkoh-eremol and agarospirol are preferentially more active than other identified compounds with strong binding affinity to major anti-inflammatory and immunomodulatory receptors. The oil displayed a significant and dose dependent reduction of 12-O-tetradecanoylphorobol-13 acetate (TPA)- induced ear edema and MDA activity when compared with vehicle treated mice. Pro-inflammatory cytokines (IL-1ß, IL-6 and TNF-α) were also reduced significantly in a dose dependent manner in all the TPA treated groups as compared to control. The present study indicates that agarwood oil significantly reduced the skin thickness, ear weight, oxidative stress and pro-inflammatory cytokines production in TPA-induced mouse ear inflammation model and contributed towards validation of its traditional use to treat inflammation related ailments.