THE FUNCTIONAL ROLE OF MELATONIN AND KISSPEPTIN IN FISH REPRODUCTION

Reproduction of fish generally governed by the counter interaction of some endogenous and exogenous cues like endocrine signals and one or more environmental factors such as photoperiod, temperature, rainfall, food availability, salinity etc. Study indicates that it depends on hypothalamo-hypophysis-gonadal axis. In gonad, gametogenesis is ordered by various hormones followed by their interactions with proper tissues leading to breed at a specific time of the year. Melatonin, a neuroendocrine hormone, has a significant physiological role in the regulation of seasonality and gonadal development of fish. On the other hand, receptor independent actions or free radical scavenging role of melatonin at different levels of hypothalamo-hypophysis-gonadal axis is not yet clear.  In recent years, focus has been given to study the of kisspeptin peptides potentiality on gonadal development especially during the transitional period of puberty and gonadal relapse. The relationship of kisspeptin and gonadotropin-releasing hormone is a good illustration of the kisspeptin system. The role of kisspeptin and melatonin in the reproduction of fish is an important subject of research.  The potency of kisspeptin and melatonin on reproductive maturation and gonadotropin release in response to environmental cues and the regulation of secretion of gonadotropin-releasing hormone by kisspeptin and melatonin are also the important objectives of the present review

IL-15 activated human peripheral blood dendritic cell kill allogeneic and xenogeneic endothelial cells via apoptosis

IL-15 is a pleotropic cytokine, which plays an important role in natural killer (NK) cell activity, T cell proliferation, and T cell cytotoxic activity. Dendritic cells (DCs) are the major antigen presenting cells in the immune system and presumed to play an important role in immune recognition of allo and xenotransplantation. We showed that IL-15 activated human peripheral blood DC is cytotoxic to human and porcine aortic endothelial cells. Unlike DCs, CD14+ monocytes show no cytotoxicity against the endothelial cells. This cytotoxic potential of IL-15 activated DC against endothelial cells is dose dependent and increases significantly upon treatment of endothelial cells with inflammatory cytokines like TNF-a or IFN-c. The cytotoxic potential of IL-15 activated DC is associated with apoptosis of endothelial cells, as indicated by the increased Annexin V staining, caspase activation and loss of mitochondrial membrane potential. Further it was observed that DC mediated cytotoxicity against endothelial cell is mediated via granzyme B possibly secreted by the activated DCs

Rhinosporidiosis: A Rare Cause of Proptosis and an Imaging Dilemma for Sinonasal Masses

Background. Rhinosporidiosis is a common disease entity in tropical countries; however, it can be encountered in other parts of the world as well due to increasing medical tourism. It may mimic other more malignant and vigorous pathologies of the involved part. Case Report. We present a case of a 36-year-old male presenting with proptosis due to involvement of nasolacrimal duct which is rare. We will discuss typical CT and MRI features of the disease which were present in the case. Conclusion. For a surgeon and a radiologist, this is a necessary differential to be kept in mind for sinonasal masses. CT and MRI are invaluable investigations. However, FNAC is confirmatory. Both clinical and radiological aspects are required to reach correct diagnosis

Targeted Delivery of Doxorubicin-Loaded Poly (ε-caprolactone)-b-Poly (N-vinylpyrrolidone) Micelles Enhances Antitumor Effect in Lymphoma

<div><p>Background</p><p>The present study was motivated by the need to design a safe nano-carrier for the delivery of doxorubicin which could be tolerant to normal cells. PCL₆₃-b-PNVP₉₀ was loaded with doxorubicin (6 mg/ml), and with 49.8% drug loading efficiency; it offers a unique platform providing selective immune responses against lymphoma.</p><p>Methods</p><p>In this study, we have used micelles of amphiphilic PCL₆₃-b-PNVP₉₀ block copolymer as nano-carrier for controlled release of doxorubicin (DOX). DOX is physically entrapped and stabilized in the hydrophobic cores of the micelles and biological roles of these micelles were evaluated in lymphoma.</p><p>Results</p><p>DOX loaded PCL₆₃-b-PNVP₉₀ block copolymer micelles (DOX-PCL₆₃-b-PNVP₉₀) shows enhanced growth inhibition and cytotoxicity against human (K-562, JE6.1 and Raji) and mice lymphoma cells (Dalton's lymphoma, DL). DOX-PCL₆₃-b-PNVP₉₀ demonstrates higher levels of tumoricidal effect against DOX-resistant tumor cells compared to free DOX. DOX-PCL₆₃-b-PNVP₉₀ demonstrated effective drug loading and a pH-responsive drug release character besides exhibiting sustained drug release performance in in-vitro and intracellular drug release experiments.</p><p>Conclusion</p><p>Unlike free DOX, DOX-PCL₆₃-b-PNVP₉₀ does not show cytotoxicity against normal cells. DOX-PCL₆₃-b-PNVP₉₀ prolonged the survival of tumor (DL) bearing mice by enhancing the apoptosis of the tumor cells in targeted organs like liver and spleen.</p></div

DOX-PCL₆₃-b-PNVP₉₀ hinders colony formation in DL.

<p>Clonogenic assay was performed in six-well plates following seeding of 100 cells with clones produced by DL tumor cells. Untreated controls and PCL₆₃-b-PNVP₉₀ treated cells formed colonies (A and C). Smaller size colonies with less number of cells were observed following doxorubicin (0.5 μM) treatment (B). Cells treated with DOX-PCL₆₃-b-PNVP₉₀ treatment (0.5 μM) unable to form colony (D). Survival analysis of DL cells treated with free DOX (Red Line) or, DOX-PCL₆₃-b-PNVP₉₀ (Blue line), (p = 0.01) (E). DOX-PCL₆₃-b-PNVP₉₀ induced growth inhibition in DL cells is caspase dependent. Growth inhibition study in presence and absence of pan caspase inhibitor ZVAD-FMK for 48 h (n = 3, mean ±SD). MTT assay was performed as described above (G).Data presented as mean ± SD, n = 3.</p

Effect of free DOX and DOX- PCL63-b-PNVP90 on normal Cell viability.

<p>Normal human lymphocytes (A), DC (B), and monocytes (C) were treated with serial concentrations of doxorubicin (0.0001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, and 5 μM). Plates were incubated at 37°C, 5% CO₂, for 48 h. The cell viability was measured by XTT assay kit (Cell Signaling, USA). Data shown as mean ± SD, n = 3. Effects on leukocytes number following in vivo administration of free DOX and DOX-PCL₆₃-b-PNVP₉₀ was studied in normal mice (D) tumor bearing mice (E) mice treated with free DOX (F), or DOX- PCL₆₃-b-PNVP₉₀ (G). RBC (H) and differential leukocyte count (I) of individual treatment are shown, mean ± SD, n = 3. (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 in control versus experimental group).</p

Induction of apoptosis by DOX-PCL₆₃-b-PNVP₉₀ micelles.

<p>The effect of free DOX or, DOX-PCL₆₃-b-PNVP₉₀ on the induction of apoptosis in parental (A) and doxorubicin resistant (DOX-R) (B) tumor cells was quantified by flow cytometric analysis of Annexin V FITC staining in tumor cells following 18 h treatment with indicated drugs. The lower left (LL) quadrant represents live/healthy cells, the lower right (LR) quadrant represents early apoptosis, and the upper right (UR) represents cells in late apoptosis, while the upper left (UL) represents the percent DOX uptake. The percentage of cells undergoing early or, late apoptosis is indicated in the respective quadrate. Data shown are mean ± SD, n = 3.</p

Time dependent uptake of free doxorubicin and DOX-PCL₆₃-b-PNVP₉₀ micelles.

<p>Uptake of free DOX or DOX-PCL₆₃-b-PNVP₉₀ micelles by parental and doxorubicin-resistant K-562 (A). JE6.1 (B) Raji (C) or, DL (D) cells were studied using fluorescence plate reader. Intracellular DOX concentration (μM) was measured in triplicate at different time points up to 30 h. Data presented as Mean ± SD, n = 4. Flow cytometric measurements of cellular DOX levels in parental K-562 (E) JE6.1 (F), Raji (G) and DL (H) (n = 3). Temporal uptake of free DOX or DOX-PCL₆₃-b-PNVP₉₀ micelles into K-562 (I) and DL (J) cells were studied by incubating in 24-well plates at a concentration of 50,000 cells per well. Representative images shown were obtained using a fluorescence microscope Eclipse 80i (Nikon, Japan) (Plan Fluor, 40X, NA 0.75 objective) equipped with blue and red filters for, Hoechst and Doxorubicin respectively (n = 3).</p