Cathepsin-K inhibition enhances anti-cancerous activity within oral squamous cell carcinoma cells: Uncloaking the potency of new K21 formulation.

BACKGROUND: The ability of cancer cells to be invasive and metastasize depend on several factors, of which the action of protease activity takes center stage in disease progression. PURPOSE/OBJECTIVE: To analyze function of new K21 molecule in the invasive process of oral squamous cell carcinoma (OSCC) cell line. MATERIALS & METHODS: The Fusobacterium (ATCC 23726) streaks were made, and pellets were resuspended in Cal27 (ATCC CRL-2095) OSCC cell line spheroid cell microplate. Cells were seeded and Lysotracker staining performed for CathepsinK red channel. Cell and morphology were evaluated using Transmission Electron microscopy. Thiobarbituric acid assay was performed. OSCC was analyzed for Mic60. Raman spectra were collected from the cancer cell line. L929 dermal fibroblast cells were used for Scratch Assay. ELISA muti arrays were used for cytokines and matrix molecules. Internalization ability of fibroblast cells were also analyzed. Structure of K21 as a surfactant molecule with best docked poses were presented. RESULTS: Decrease in lysosomal staining was observed after 15 and 30 min of 0.1% treatment. Tumor clusters were associated with cell membrane destruction in K21 primed cells. There was functional silencing of Mic60 via K21, especially with 1% concentration with reduced cell migration and invasiveness. Raman intensity differences were seen at 700 cm-1, 1200 cm-1 and 1600 cm-1 regions. EVs were detected within presence of fibroblast cells amongst K21 groups. Wound area and wound closure showed the progress of wound healing. CONCLUSION: Over expression of CatK can be reduced by a newly developed targeted K21 based drug delivery system leading to reduced migration and adhesion of oral squamous cell carcinoma cells. The K21 drug formulation can have great potential for cancer therapies due to targeting and cytotoxicity effects.

Pre-weaning fluoxetine exposure perturbs social behavior at adulthood via altering hippocampal morphometry and PSD-95 expression in rats.

The depression during and after pregnancy cause significant exposure of fluoxetine to the child at early life through mother. This exposure to the child, during the vulnerable window of development, can have a long lasting impact on overall mental wellbeing. Long term neurobehavioral aspect of developmental toxicity is neglected as the part of testing requirements in the process of drug developmental. In this context, the present study was designed to study the possible effect of pre-weaning fluoxetine exposure on the social behavior of rats upon adulthood followed by assessing hippocampal morphometry (hematoxylin-eosin and silver staining) and post-synaptic density protein 95 (PSD-95) expression (using qPCR). Our data showed that the fluoxetine exposure (10, 50 and 100mg/kg) caused predominant increase in the social behavior of rats; the effect more pronounced in female rats. The morphometric analysis revealed significant increase in cell population and count of dentate gyrus (DG) region of hippocampus along with enhanced dendritic arborization. Furthermore, the PSD-95 expression was found to be down regulated in the fluoxetine treated group as compared to control. In conclusion, the present study demonstrate that the early post-natal exposure to fluoxetine cause hypersociability upon attaining adulthood, which may be attributed to enhanced neuronal proliferation and decrease PSD-95 expression in the hippocampus.

Attenuation of cisplatin-induced acute kidney injury by N-(2-Hydroxyphenyl) acetamide and its gold conjugated nano-formulations in mice.

The attenuation of cisplatin-induced acute kidney injury (AKI) in mice by N-(2-hydroxyphenyl) acetamide (NA-2) and NA-2-conjugated gold nanoparticles (NA2-AuNPs) was investigated. Male BALB/c mice (n = 54) were divided into nine groups having six animals in each group. Animals in groups 3-9 were pre-treated for 5 days with test compounds, whereas, animals in group 1 and 2 received normal saline. On day 4, animals in groups 2, 3, 4, 5, 6 and 9 were given single intra-peritoneal injection of CP at the dose of 5 mg/kg. After 72 hours of CP injection, all animals were sacrificed. Blood was collected for serum urea and creatinine estimation, and kidneys were harvested for histo-pathological examinations and qPCR studies for nuclear factor-κB p50, (NFκB) ; inducible nitric oxide synthase (iNOS); hemeoxygenase-1 (HO-1); and interleukin-6 (IL-6).NA-2 and NA2-AuNPs was observed to decrease the serum urea and creatinine levels. Both the test compounds reduced kidney injury damage score and improved histological architecture in the treated animals in dose dependent manner. Furthermore, the mRNA expressions of NFkB p50, iNOS and IL-6 genes were down-regulated, and HO-1 gene was up-regulated in the animals treated with the test compounds. It is concluded that NA-2 and NA2-AuNPs attenuates CP-induced AKI in mice models through anti-inflammatory and anti-oxidant mechanisms.

Rutin coated gold nanoparticles prevent rhabdomyolysis-induced kidney injury via down-regulation of NF-kB, iNOS, IL-6 and up-regulation of HO-1 and Kim-1 genes in mice.

The aim of this study was to evaluate the protective activity of rutin, and its gold nanoparticles (Ru-AuNPs) in rhabdomyolysis-induced acute kidney injury (AKI) model in mice. Rutin (25 and 50 mg/kg) and Ru-AuNPs (15 and 25 mg/kg) were administered to the animals for four (4) days with water deprivation for 24 hours followed by 50% glycerol injection at the dose of 10 ml/kg intramuscularly. On the next day, animals were dissected and blood and kidneys were collected. Biochemical investigations were performed to evaluate kidney functions, histological studies were carried out to see the changes at tissue level and real-time RT-PCR studies for nuclear factor-κB p50, NFκB; inducible nitric oxide synthase, iNOS; heme oxygenase-1, HO-1; interleukin-6, IL-6; and kidney injury molecule-1, Kim-1 were performed to elucidate the molecular mechanisms. Blood urea and creatinine were found to be decreased in animals treated with rutin and Ru-AuNPs. Down regulation of the mRNA expressions of iNOS, IL-6 and NFkB p50 and up-regulation of Kim-1 and HO-1 genes were observed. The efficacy of Ru-AuNPs was better than rutin alone even at a dose far less than the compound. Rutin and Ru-AuNPs alleviates kidney injury and inflammation in rhabdomyolysis-induced AKI model via anti-inflammatory and anti-oxidant pathways which make it a plausible compound for future studies.

Eugenol and liposome-based nanocarriers loaded with eugenol protect against anxiolytic disorder via down regulation of neurokinin-1 receptors in mice.

Anxiety disorder is a psychiatric disorder characterized by extreme fear or worry. It is highly prevalent worldwide which affects daily life and is also an enormous health burden. Neurokinin 1 receptor (NK1R) is a G protein coupled receptor, expressed in both central and peripheral nervous system, involved in affective behaviors. NK1R has established role in anxiety and it is also an important target for pathogenesis of anxiety disorder. Therefore, it has been hypothesized in previous studies that the blockades of NK1R may have antidepressant and anxiolytic effects. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of NK1R was analyzed by real time RT-PCR. Moreover, the NK1R protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating NK1R protein expression in mice.

N-(2-hydroxyphenyl)acetamide and its gold nanoparticle conjugation prevent glycerol-induced acute kidney injury by attenuating inflammation and oxidative injury in mice.

The protective activity of N-(2-hydroxyphenyl)acetamide (NA-2) and NA-2-coated gold nanoparticles (NA-2-AuNPs) in glycerol-treated model of acute kidney injury (AKI) in mice was investigated. NA-2 (50 mg/kg) and NA-2-AuNPs (30 mg/kg) were given to the animals for four days followed by 24-h water deprivation and injection of 50% glycerol (10 ml/kg im). The animals were sacrificed on the next day. Blood and kidneys were collected for biochemical investigations (urea and creatinine), histological studies (hematoxylin and eosin; and periodic acid-Schiff staining), immunohistochemistry (actin and cyclooxygenase-2, Cox-2), and real-time RT-PCR (inducible nitric oxide synthase, iNOS; nuclear factor-κB p50, NFκB; hemeoxygenase-1, HO-1; and kidney injury molecule-1, Kim-1). NA-2 protected renal tubular necrosis and inflammation, though the result of NA-2-AuNPs was better than compound alone and it also exhibited the activity at far less dose. The test compound and its gold nano-formulation decreased the levels of serum urea and creatinine level in the treated animals. Both NA-2 and NA-2-AuNPs also conserved actin cytoskeleton, and lowered COX-2 protein expression. Moreover, the mRNA expressions of iNOS and NFkB p50 were down-regulated, and HO-1 and Kim-1 genes were up-regulated. We conclude that NA-2 and NA-2-AuNPs ameliorates kidney inflammation and injury in glycerol-induced AKI animal model via anti-oxidant and anti-inflammatory mechanisms which make it a suitable candidate for further studies. We believe that these findings will contribute in the understanding of the mechanism of action of paracetamol-like drugs and can be considered for clinical research for the prevention of AKI.