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Objective In attempt to conquer the major concerns of oral duloxetine hydrochloride (like low bioavailability, intolerable side-effects and no regeneration of demyelinated nerve fibres) for the management of chemotherapy-induced peripheral neuropathy (CIPN), an alternative delivery of duloxetine hydrochloride was aimed for in-vivo optimization. Methods A film forming dermal gel consisting of duloxetine hydrochloride was formulated and enriched with methylcobalamin and geranium oil. The formulated gel successfully qualified the various pharmaceutical characteristics of gel. Administration of paclitaxel (8 mg/kg/i.p. in four divided doses) for 4 alternate days induced the symptoms of peripheral neuropathy in rats. On 14th day, the responses to noxious stimulus (mechanical hyperalgesia, cold allodynia, and heat hyperalgesia) were increased and reached to its maximum. Thereafter, drug treatment with formulated dermal gel and oral duloxetine hydrochloride (30 mg/kg, once daily) was initiated for 2 weeks in different group of animals. On the 28th day animals were sacrificed to isolate sciatic nerve, to assess biochemical changes (TBARS, reduced GSH, total protein, TNF-α, IL-6) and for histopathological examinations of nerve sections using Hematoxylin-Eosin and Toludine blue staining methods. Results Application of formulated dermal gel to paclitaxel-treated rats significantly improved paw-withdrawal latency responses during noxious stimulus testing, reduced the levels of TBARS, TNF-α, IL-6 and elevated the levels of reduced GSH as compared to paclitaxel treated rats. Histographs also indicated marked regeneration of the damaged nerve fibers. Topical delivery of duloxetine hydrochloride produced similar results in disparity to oral route. However, no significant disparity in responses was obtained with twice application of formulated dermal gel when compared to once daily application. Conclusion Tremendous recovery from nociception, oxidation and inflammation in addition to nerve degeneration was achieved through dermal application of duloxetine hydrochloride in peripheral neuropathy.

Protocols are set of predefined written procedural documents that offer guidance or specifications for performing the experiments and its implementation. They are an essential component of good laboratory practice (GLP) and good clinical practice (GCP) regulations that establish the study’s rationale, objectives, design, and planned analyses of the results, as well as the conditions under which these procedures will be carried out. In addition to detailed procedures, protocols often include information on safety precautions, the calculation of results, and reporting standards, including statistical analysis and rules for predefining and documenting excluded data to avoid bias. The protocol is the heart of every experiment. It is the plan; it is a critical document for everyone involved in the conduct of the procedure—Rebecca Kush, President of CDISC Different protocols are available which provide signaling pathways of introduction to “Programmed cell death” which called “Apoptosis”. The course of programmed cell death or “Apoptosis” is generally regarded as a typical element of the human health and disease state. Normal homeostasis is sustained throughout by maintaining balance between cell death and cell division. Apoptosis is phenomenon that involves specific activation of death-signaling pathways leading to the removal of cells from tissue. The individual morphological features of apoptosis include cell shrinkage, chromatin condensation, membrane blebbing, and formation of apoptotic bodies. Inapt apoptosis is a key factor in many diseased conditions, including neurodegenerative diseases, cancer, I-R injury, and many more. In addition, the area of apoptosis research is growing at an alarming rate and till date vast research has been made on the clarification and analysis of the cell cycle mechanisms and signaling pathways regulating cell cycle. The aptitude to detect and quantify apoptosis and to understand its biochemistry related to regulatory genes and proteins is highly crucial for biomedical research. This chapter is an attempt to highlight various basic as well as advanced tools and techniques (like immunohistochemistry, TUNEL assay, in-situ end-labeling techniques in conjunction with standard flow cytometry) used in area of apoptosis research in addition to focus on protocols used in qualitative and quantitative determination of apoptosis to be followed during preclinical and clinical phase of drug development. The use of aforementioned techniques for apoptosis quantification will enable clinical investigators to accurately assess apoptosis in context of various diseases.

Pregnane X receptors (PXRs) regulate the expression of ATP-binding cassette proteins transporters and organic anion transporting polypeptides responsible for influx/efflux of xenobiotics across the brain. Ligand activation of PXR augments the expression of P-gp and promotes amyloid-β clearance across the blood–brain barrier. Dementia was induced in mice by intacerebroventricular administration of streptozotocin (STZ) followed by treatment with meclizine, a PXR agonist, and subsequently exposed to the Morris water maze test and biochemical and histopathological analysis to evaluate the effect on cognition. STZ-treated mice exhibited significant enhancement in brain thiobarbituric acid reactive species, interleukin-1β, tumour necrosis factor-α, myeloperoxidase, and acetylcholinestrase activity in addition to diminution in glutathione levels and superoxide dismutase activity in comparison to untreated mice. Administration of meclizine to STZ mice recuperated cognition and biochemical alterations. Concomitant administration of ketoconazole, a PXR antagonist, with meclizine prevented the protective effects. The upshots of our study proclaim that meclizine protects cognitive deficits by virtue of its antioxidant, anticholinesterase, and antiinflammatory properties. Results also signify the potential of PXR in neuroprotective actions of meclizine in dementia.

Background Studies have signified that high serum cholesterol plays an intriguing role in amyloid β metabolism and accumulation. Ligand activation of pregnane x receptors (PXRs), up-regulates the expression of P- glycoprotein and has a crucial role in amyloid β efflux. The present study has been undertaken to investigate the effect of forskolin, a PXR agonist in experimental dementia. Methods Wistar rats were allowed free access to cholesterol-rich High Fat Diet (HFD) for 90 days to induce dementia. HFD rats were then treated with forskolin (10 mg/kg; 20 mg/kg) followed by exposure to Morris water maze (MWM) test to deconvolute the mechanistic of learning and memory. An array of biochemical and histopathological tests were performed to demonstrate the extent of damage induced by HFD. Results HFD-treated rats exhibited marked accentuation in brain thiobarbituric acid reactive species, Interleukin-1β, tumor necrosis factor-α levels, myeloperoxidase and acetylcholinestrase activity in addition to attenuation of glutathione levels and superoxide dismutase activity as compared to rats fed on normal chow diet. Consistent rise in serum cholesterol level was also indicated. Histopathological examination of cerebral cortex using hematoxylin and eosin and congo red staining methods demonstrated significant neutrophilic incursion and amyloid deposition. Administration of forskolin to HFD treated rats improved memory functions, biochemical and histopathological alterations. Concomitant administration of ketoconazole, a PXR antagonist with forskolin prevented the observed protective effects. Conclusion Our findings signify that forskolin defends HFD induced cognitive deficits. Current plethora of results also defines the potential of PXR in neuroprotective action of forskolin in dementia.