Granulocyte colony stimulating factor (GCSF) can decrease mortality of patients undergo chemotherapy through increasing neutrophil counts. Many strategies have been developed to improve its blood circulating time. Albumin binding domain (ABD) was genetically fused to N-terminal end of GCSF encoding sequence and expressed as cytoplasmic inclusion bodies within Escherichia coli. Biological activity of ABD-GCSF protein was assessed by proliferation assay on NFS-60 cells. Physicochemical properties were analyzed through size exclusion chromatography, circular dichroism, intrinsic fluorescence spectroscopy and dynamic light scattering. Pharmacodynamics and pharmacokinetic properties were also investigated in a neutropenic rat model. CD and IFS spectra revealed that ABD fusion to GCSF did not significantly affect the secondary and tertiary structures of the molecule. DLS and SEC results indicated the absence of aggregation formation. EC50 value of the ABD-GCSF in proliferation of NFS-60 cells was 75.76 pg/ml after 72 h in comparison with control GCSF molecules (Filgrastim: 73.1 pg/ml and PEG-Filgrastim: 44.6 pg/ml). Animal studies of ABD-GCSF represented improved serum half-life (9.3 ± 0.7 h) and consequently reduced renal clearance (16.1 ± 1.4 ml/h.kg) in comparison with Filgrastim (1.7 ± 0.1 h). Enhanced neutrophils count following administration of ABD-GCSF was comparable with Filgrastim and weaker than PEG-Filgrastim treated rats. In vitro and in vivo results suggested the ABD fusion as a potential approach for improving GCSF properties.
Background: Pomegranate (Punica granatum) is a significant source of bioactive compounds. However, its toxicity is not intensively studied. Objectives: The current study investigated the safety and tolerability of pomegranate peel extract (PPE) in BALB/c mice. Materials and Methods: A total of 25 female BALB/c mice were randomly grouped. Each experimental group consisted of five animals. Repeated doses including 0.5, 1.9 and 7.5 mg/kg body weight of PPE were gavaged to BALB/c mice, for 22 days and the single intra-dermal injection (224 mg/kg) was done in one dose. The control group administrated with distilled water was also included. In addition, intra dermal injection for skin allergy testing was also performed. Blood was collected to evaluate glucose, cholesterol, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) as indicators of liver toxicity. Macroscopic and histopathological evaluation of tongue, trachea, and larynx tissues were also performed on 22 days post administration. Results: Toxicological potential of PPE studies revealed no toxic effects, clinical signs, histopathological effect in epithelial cells layer of tongue, larynx and trachea, behavioral alterations and adverse effects or mortality in BALB/c mice. Repeated administrations did not alter or cause local irritation of the oral mucosa. Skin allergy test was negative in the last group. Conclusions: The current study showed that PPE had no toxicity and its use is suggested with potential applications against diseases.
Background and Objectives: Cholera disease remains an important global health problem affecting 3-5 million subjects worldwide. Outer membrane vesicles (OMVs) have been found in a variety of Gram-negative bacteria and act as protective transport vesicles. The aim of this study was to evaluate Immune responses against Vibrio cholerae O1 El Tor clinical strain OMV and compare it with killed whole cell (KWC), complex of (KWC-OMV) as well as the internationally licensed oral cholera vaccine, Dukoral, in serum and intestinal secretions of mice. Materials and Methods: OMVs were prepared by using modified detergent-centrifugation procedure from V. cholerae O1 El Tor clinical strain from 2005 outbreak. The ultrastructure and content of OMVs were investigated via the Scanning Elec- tron Microscopy (SEM) and SDS-PAGE analysis. Three doses of oral immunization were adjusted and total IgG and IgA in serum and intestinal secretion were measured by enzyme-linked immunosorbent assay (ELISA). Results: Extracted OMVs from the V. cholerae were spherical vesicles with a size ranging from 10 to 300 nm. OMV-im- munized mice showed an increased level of total IgG and IgA both in serum and intestinal secretion when compared to the negative controls. Also, there existed a higher level of secretory IgA than the total IgG, suggesting the most of protection against V. cholerae colonization provided by sIgA. Conclusion: Our findings revealed that oral immunization with V. cholerae OMVs might induce a long-term immunity, es- pecially when administered in combination with KWC. This study tested the adjuvant activity of OMVs and may be useful in future nano vaccine research.
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