Morphological Modification of Carbon Nanoparticles after Interacting with Methotrexate as a Potential Anticancer Agent

Purpose

Production of highly penetrable and targetable drug delivery particles is mainly focused by current therapy and such focus is achieved in our present study. The carbon nanoparticle (CNP) prepared from purely natural source was modified from spherical shape to cylindrical floral like structure after treatment with the anticancer drug methotrexate (CM).

Methods

The physiochemical properties of the CNP and CM was characterized using FT-IR/Raman Spectrometer, XRD, SEM, AFM, particle size analyzer and its biological evaluation using haemolysis and MTT assay.

Results

The shift in FT-IR peaks at 1592, 1120 cm^?1 and peaks of raman spectra observed at 1303, 1300 cm^?1 represents ordered carbon nanotubes. The morphological change from spherical to cylindrical floral like structure was observed using SEM and AFM and its particle size distribution analysis shows an average diameter of 269 nm for CM. XRD peak at 2θ?=?23.86° (002) indicates the presence of large amount of amorphous material that corresponds to multi-walled carbon nanotubes. Haemocompatibility studies proved the safety level usage as 100 μg/ml and MTT assay shows viability rate of 85–98% with mouse embryonic fibroblast (NIH/3 T3) and 30–45% with pancreatic carcinoma (MIA PaCa-2) and gastric cancer cell lines (SNU- 484) respectively.These results are also supported by phase contrast microscope images observed after staining with calcein AM and EthD-1.

Conclusions

The morphologically modified CNPs has shown good anticancer, biocompatibility and haemocompatibility property which is an important criterion to be satisfied by a biomedical product.

     

Extracorporeal shock wave therapy induces alveolar bone regeneration

Periodontal inflammation with alveolar bone resorption is a hallmark of periodontitis. We hypothesized that extracorporeal shock wave therapy (ESWT) could promote the regeneration of alveolar bone following Porphyromonas gingivalis-induced periodontitis in rats. Rats were infected with P. gingivalis for 10 wks, which caused alveolar bone resorption. The rats were then treated with a single episode of 100, 300, or 1000 impulses of shock wave on both cheeks at energy levels 0.1 mJ/mm(2). Alveolar bone levels were determined at 0, 3, 6, and 12 wks following ESWT and compared with those in untreated controls. Infected rats treated with 300 and 1000 impulses demonstrated significantly improved alveolar bone levels at 3 wks compared with untreated controls, and the improved levels remained for at least 6 wks in most rats. The results demonstrated effective regeneration of alveolar bone by ESWT and suggested that ESWT should be evaluated as an adjunct in the regeneration of periodontal tissues following periodontal disease. Abbreviations: ESWT, extracorporeal shock wave therapy; PCR, polymerase chain-reaction.     

Highly sensitive naphthalimide based Schiff base for the fluorimetric detection of Fe3+

A simple 1,8-naphthalimide based Schiff base probe (E)-6-((4-(diethylamino)-2-hydroxybenzylidene)amino)-2-(2-morpholinoethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (NDSM) has been designed and synthesized for the specific detection of Fe³⁺ based on a fluorimetric mode. The absorbance of NDSM at 360 nm increased significantly in acetonitrile : water (7 : 3, v/v) medium only in the presence of Fe³⁺ ions with a visible colour change from yellow to golden yellow. Likewise, fluorescence emission intensity at 531 nm was almost wholly quenched in the presence of Fe³⁺. However, other competitive ions influenced insignificantly or did not affect the optical properties of NDSM. Lysosome targetability was expected from NDSM due to the installation of a basic morpholine unit. The LOD was found to be 0.8 μM with a response time of seconds. The fluorescence reversibility of NDSM + Fe³⁺ was established with complexing agent EDTA. Fe³⁺ influences the optical properties of NDSM by complexing with it, which blocks C N isomerization in addition to the ICT mechanism. The real-time application of Fe³⁺ was demonstrated in test paper-based detection, by the construction of a molecular logic gate, quantification of Fe³⁺ in water samples and fluorescence imaging of Fe³⁺.

A simple 1,8-naphthalimide based Schiff base probe NDSM has been designed and synthesized for the specific detection of Fe³⁺ based on a fluorimetric mode.     

Inhibition of IKKβ by celastrol and its analogues – an in silico and in vitro approach

Context: Alzheimer’s disease (AD) is the most common form of dementia affecting the aged population and neuroinflammation is one of the most observed AD pathologies. NF-κB is the central regulator of inflammation and inhibitor κB kinase (IKK) is the converging point in NF-κB activation. Celastrol is a natural triterpene used as a treatment for inflammatory conditions.

Objective: This study determines the neuroprotective and inhibitory effect of celastrol on amyloid beta_1-42 (Aβ_1-42) induced cytotoxicity and IKKβ activity, respectively.

Materials and methods: Retinoic acid differentiated IMR-32 cells were treated with celastrol (1?μM) before treatment with Aβ_1-42 (IC₃₀ 10?μM) for 24?h. The cytotoxicity and IKK phosphorylation were measured by MTT and western blotting analysis, respectively. We screened 36 celastrol analogues for the IKKβ inhibition by molecular docking and evaluated their drug like properties to delineate the neuroprotective effects.

Results: Celastrol (1?μM) inhibited Aβ_1-42 (10?μM) induced IκBα phosphorylation and protected IMR-32 cells from cell death. Celastrol and 25 analogues showed strong binding affinity with IKKβ as evidenced by strong hydrogen-bonding interactions with critical active site residues. All the 25 analogues displayed strong anti-inflammatory properties but only 11 analogues showed drug-likeness. Collectively, molecule 15 has highest binding affinity, CNS activity and more drug likeness than parent compound celastrol.

Discussion and conclusion: The decreased expression of pIκBα in celastrol pretreated cells affirms the functional representation of inhibited IKKβ activity in these cells. The neuroprotective potentials of celastrol and its analogues may be related to IKK inhibition.     

Two cases of CARD14-associated papulosquamous eruption from India

Identification of CARD14-associated papulosquamous eruption (CAPE) is important as it helps in determining prognosis and management of those affected. We report two siblings with genetically confirmed CAPE presenting with treatment-resistant erythroderma in one patient and patterned psoriatic plaques with facial predominance in the other.     

Structure‐Based Identification of Aporphines with Selective 5‐HT2A Receptor‐Binding Activity

Selective blockade of the serotonin 5‐HT_2A receptor is a useful therapeutic approach for a number of disorders, including schizophrenia, insomnia and ischaemic heart disease. A series of aporphines were docked into a homology model of the rat 5‐HT_2A receptor using AutoDock. Selected compounds with high in silico binding affinities were screened in vitro using radioligand‐binding assays against rat serotonin (5‐HT_1A and 5‐HT_2A) and dopamine (D1 and D2) receptors. (R)‐Roemerine and (±)‐nuciferine were found to have high affinity for the 5‐HT_2A receptor (K_i = 62 and 139 nm , respectively), with (R)‐roemerine showing 20‐ to 400‐fold selectivity for the 5‐HT_2A receptor over the 5‐HT_1A, D1 and D2 receptors. Investigation into the ligand–receptor interactions suggested that the selectivity of (R)‐roemerine is due to it having stronger H‐bonding and dipole–dipole interactions with several of the key residues in the 5‐HT_2A receptor‐binding site.     

Sustainable pectin fascinating hydroxyapatite nanocomposite scaffolds to enhance tissue regeneration

The preparation of biocompatible nanomaterials is one of the emerging areas and it is continuously developing with the use of various contrived methods to accomplish the formation of nanoscale materials. Nevertheless, unfortunately, many of these strategies utilize harmful organic solvents, which make the pertinence of nanoparticles in medicinal applicationsimpractical. In this study, the morphology-focused hydroxyapatite (HAP) was prepared using pectin extracted from the citrus fruit peel (Citrus limonum) and it is used for the synthesis of nano HAP by varying the concentration of pectin as a template. The chemical structure, crystallinity, and morphology were determined by FTIR, XRD, and SEM, respectively. To increase the biocompatibility of HAP, pectin aided HAP (tHAP) and HAP/pectin composites were synthesized with different concentrations of pectin. The compatibility of HAP/pectin was carried out in a human osteoblast cell line. The physic-chemical and biocompatibility showed, HAP/pectin, and HAP/pectin composites are promising materials for bone tissue engineering applications.     

BAY 41-2272 [5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]pyrimidin-4-ylamine]-induced dilation in ovine pulmonary artery: role of sodium pump

The mechanisms of relaxation to nitric oxide (NO)-independent soluble guanylyl cyclase (sGC) activator BAY 41-2272 [5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]pyrimidin-4-ylamine] were investigated in isolated ovine pulmonary artery. BAY 41-2272 (1 nM-10 microM) produced concentration-dependent relaxation of endothelium-denuded pulmonary artery rings (pD2 = 6.82 +/- 0.16; Emax = 92.30 +/- 2.31%; n = 8), precontracted with 1 microM 5-hydroxytryptamine (serotonin). 1-H-[1,2,4]Oxadiazole[4,3-a]quinoxalin-1-one (ODQ; 10 microM), an inhibitor of sGC, partially inhibited (Emax = 57.10 +/- 3.10%; n = 6) the relaxation response of BAY 41-2272. In comparison with ODQ, sodium pump inhibitor ouabain (1 microM) produced a greater decrease in the vasodilator response of BAY 41-2272 (Emax = 20.17 +/- 4.55%; n = 6). K+-free solution also attenuated (Emax = 39.97 +/- 3.52%; n = 6) BAY 41-2272-induced relaxation. ODQ (10 microM) plus 1 microM ouabain abolished the relaxant response of BAY 41-2272 (Emax = 12.09 +/- 3.76%, n = 6 versus vehicle control dimethyl sulfoxide; Emax = 15.83 +/- 1.72%, n = 6). KT-5823 [1-oxo-9.12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-I][1,6]benzodiazocine-10-carboxylic acid methyl ester (2 microM), a specific inhibitor of protein kinase G had no effect on 10 microM ODQ-insensitive relaxation evoked by BAY 41-2272. BAY 41-2272 (10 microM) inhibited Ca2+-induced contractions in K+-depolarized preparations. BAY 41-2272 (10 microM) caused about a 14-fold increase in the intracellular cGMP over the basal level, which was completely inhibited by 10 microM ODQ. BAY 41-2272 (0.1, 1.0, and 10 microM) significantly (P < 0.05) increased ouabain-sensitive 86Rb uptake in a concentration-dependent manner. BAY 41-2272 (10 microM) also stimulated sarcolemmal Na+-K+-ATPase activity. However, 10 microM ODQ had no significant effect on either basal or BAY 41-2272-stimulated 86Rb uptake/Na+-K+-ATPase activities. In conclusion, this study provides the first evidence of sodium pump stimulation by BAY 41-2272 independent of cGMP as an additional mechanism to sGC activation in relaxation of ovine pulmonary artery.