Evaluation of behavioural and antioxidant activity of Cytisus scoparius Link in rats exposed to chronic unpredictable mild stress

<p>Abstract</p> <p>Background</p> <p>Various human diseases have oxidative stress as one of their component. Many herbs have been reported to exhibit properties that combat oxidative stress through their active constituents such as flavonoids, tannins, phenolic compounds etc. <it>Cytisus scoparius </it>(CS) Link, (Family: Leguminosae), also called <it>Sarothamnus scoparius</it>, has been shown in <it>invitro </it>experiments to be endowed with anti-diabetic, hypnotic and sedative and antioxidant activity. Therefore this study was carried out to evaluate CS for its anxiolytic, antidepressant and anti-oxidant activity in stressed rats.</p> <p>Methods</p> <p>60% methanolic extract of CS was quantified for phenolic content by Folin-Ciocalteau's method. Chronic unpredictable mild stress (CMS) was employed to induce stress in rats. CS (125 and 250 mg/kg, p.o) and diazepam (DZM) (2 mg/kg, p.o) was administered during the 21 day stress exposure period. Anxiolytic and antidepressant activities of CS were assessed in open field exploratory and behavioural despair paradigms, respectively. Plasma glucose and total lipids; endogenous antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT); non-enzymic-ascorbic acid and thiobarbituric acid reactive substances (TBARS) levels were measured in brain, kidneys and adrenals using standard protocols to assess the effect of CS.</p> <p>Results</p> <p>Total phenolic content of CS was found to be 8.54 ± 0.16% w/w. CMS produced anxiogenic and depressive behaviour in experimental rats with metabolic disturbance. Significant decrease in SOD, CAT levels and increase in lipid peroxidation level was observed in stressed rats. CS administration for 21 days during stress exposure significantly increased the ambulatory behaviour and decreased the freezing time in open field behaviour. In behavioural despair test no significant alteration in the immobility period was observed. CS also improved SOD, CAT, and ascorbic acid level and controlled the lipid peroxidation in different tissues.</p> <p>Conclusion</p> <p>CS possesses anti-stress and moderate anxiolytic activity which may be due, in part, to its antioxidant effect that might warrant further studies.</p

Functional and molecular characterization of hyposensitive underactive bladder tissue and urine in streptozotocin-induced diabetic rat

Background: The functional and molecular alterations of nerve growth factor (NGF) and Prostaglandin E2 (PGE2) and its receptors were studied in bladder and urine in streptozotocin (STZ)-induced diabetic rats. Methodology/Principal Findings: Diabetes mellitus was induced with a single dose of 45 mg/kg STZ Intraperitoneally (i.p) in female Sprague-Dawley rats. Continuous cystometrogram were performed on control rats and STZ treated rats at week 4 or 12 under urethane anesthesia. Bladder was then harvested for histology, expression of EP receptors and NGF by western blotting, PGE2 levels by ELISA, and detection of apoptosis by TUNEL staining. In addition, 4-hr urine was collected from all groups for urine levels of PGE2, and NGF assay. DM induced progressive increase of bladder weight, urine production, intercontraction interval (ICI) and residual urine in a time dependent fashion. Upregulation of Prostaglandin E receptor (EP)1 and EP3 receptors and downregulation of NGF expression, increase in urine NGF and decrease levels of urine PGE2 at week 12 was observed. The decrease in ICI by intravesical instillation of PGE2 was by 51% in control rats and 31.4% in DM group at week 12. Conclusions/Significance: DM induced hyposensitive underactive bladder which is characterized by increased inflammatory reaction, apoptosis, urine NGF levels, upregulation of EP1 and EP3 receptors and decreased bladder NGF and urine PGE2. The data suggest that EP3 receptor are potential targets in the treatment of diabetes induced underactive bladder. © 2014 Nirmal et al

TEM micrographs showing endocytosis mediated uptake of liposome encapsulated gold marker at 37°C (Panel A&C).

<p>Higher magnification image of inscribed area from panel A showed that vesicle like structures in endosomal compartment (marked by E) contained cluster of electron dense dark particles inside a single cell (Panel C). In contrast, only extracellular binding of liposomes containing dark grains was observed at 4°C (Panel B) and corresponding higher magnification image (Panel D) showed that vesicle like structures in a single cell were devoid of dark gold particles, which indicates absence of internalization due to temperature dependent inhibition of endocytosis. Gold encapsulated in liposomes is marked by red colored G inside a red circle, nucleus is marked by N, endocytic vesicles are marked by E and finger like projections called as lateral interdigitation are marked by LI. Inscribed area of panel A and B in white rectangle is magnified further in panel C and D, respectively and magnification is shown by scale bar in each panel.</p

TEM micrographs showing endocytosis mediated uptake of liposome encapsulated gold in rat bladder (A) as revealed by electron dense dark grains consistent with uptake of gold across the urothelium.

<p>(B) Dense black grains binding to the cell surface were noted in rat group instilled with colloidal gold in absence of liposomes. Gold encapsulated in liposomes are marked by red colored G inside a red circle and plain gold is marked by yellow G in the images. (C) Untreated rat bladder is marked by (absence of dark black gold grains. Magnification of 8900x was used in all the images and is shown by the scale bar.</p

Illustration depict temperature dependent endocytoic uptake of liposome encapsulated gold (Lipo-gold).

<p>The cellular process of clathrin and caveolin mediated endocytosis are energy dependent and therefore only occur at 37°C and are inhibited at 4°C. Compared to mhCD, chlorpomazine was more efficient in blocking endocytosis of liposomes, which indicates a predominance of clathrin mediated endocytosis as a mechanism of endocytosis.</p

Effect of different endocytic inhibitors on cellular uptake of fluorescent liposomes.

<p>Flow cytometry analysis suggest clathrin mediated endocytosis as the dominant pathway for the internalization of fluorescently labeled liposomes by UROtsa cells at 37°C. Chlorpromazine (inhibitor of clathrin) dose dependently inhibited the internalization as cell fluorescence from externally bound non-internalized liposomes was quenched by sodium dithionite. Fluorescence intensity of liposomes in cells untreated with inhibitors was taken as 100%.</p

Drug, delivery and devices for diabetic retinopathy (3Ds in DR)

Introduction: Diabetic Retinopathy (DR) is one of the most common causes of blindness among the working population worldwide. Clearly, there is an unmet clinical need to find better treatment options for DR. Areas covered: The literature search was conducted on PubMed with no limitation on language or year of publication. The review focuses on the clinically used drugs/proteins along with a brief background on the pathophysiology of DR. The major focus of this review revolves around three treatment approaches involving drugs/proteins, drug delivery formulations and drug delivery devices. In each category, major advances are discussed along with the possible solutions. We have also discussed the various modes of administration that are currently being evaluated in the clinic. An attempt has been made to address the potential targeted site of action for DR drug delivery, and also to understand the role of Blood Retinal Barrier (BRB). Expert Opinion: In the current scenario, although there have been some advances in the drug delivery devices for delivering drugs/proteins, there are still challenges to be overcome with regard to the particulate systems. For long-term success of DR therapeutics, research options should consider taking into account the 3Ds (drug, delivery and devices).Accepted versio

A review of the role of intravitreal corticosteroids as an adjuvant to antibiotics in infectious endophthalmitis

Infectious endophthalmitis is an important cause of vision loss worldwide. This entity most often occurs as a complication of intraocular surgery especially following cataract surgery or intravitreal injection. Endophthalmitis is regarded as a serious complication following ocular surgery and the final visual outcome is fundamentally contingent on timely recognition and intervention. Intravitreal and oral antibiotics in combination with pars plana vitrectomy or vitreous aspiration remain the mainstay in the management of endophthalmitis. However, significant inflammation may persist even after sterilization of the intraocular cavities with appropriate antibiotics resulting in failure of treatment. This forms the basis for the use of intravitreal corticosteroids as an adjuvant to antibiotics in the management of infectious endophthalmitis. In the index manuscript, we review the existing literature to determine the role of intravitreal corticosteroids as an adjuvant to antibiotics in treating infectious endophthalmitis, and discuss their beneficial effects and controversial concerns