High expression level of human epidermal growth factor (hEGF) using a well-designed fusion protein-tagged construct in E. coli.

AIM: The study was aimed at design a good fusion construct that would successfully express the recombinant proteins and produce peptides in Escherichia coli. Two different constructs including human epidermal growth factor (hEGF) gene were designed to obtain an efficient expression level of hEGF. The hEGF sequence was inserted in pET32a vector containing thioredoxin (Trx) sequence and modified pET15b vector containing intein and elastin-like polypeptide (ELP). METHODS: The vectors were transformed into E. coli TOP10F' for multiplication and further into E. coli BL21 (DE3) to express protein. The hEGF expression was induced by isopropyl ß-D-1-thiogalactopyranoside (IPTG) while the expression levels were evaluated by SDS-PAGE and western blotting and compared by ImageJ analysis, BCA and Elisa assays. RESULTS: The expression level after 2 hours of IPTG induction was significantly higher than after other induction times. ImageJ, BCA and Elisa analyses demonstrated that the Trx presence enhanced protein expression significantly when compared to ELP-intein-based construct. CONCLUSION: The pET32a-Trx-hEGF construct had a higher expression than pET15b-ELP-intein-hEGF. Overall, considering Trx, the fusion protein in construct design can make it suitable to significantly express hEGF compared to ELP-intein while its combination with ELP-intein may improve the expression of the ELP-intein construct (Tab. 2, Fig. 7, Ref. 34).

SERUM NESFATIN-1 LEVEL IN HEALTHY SUBJECTS WITH WEIGHT-RELATED ABNORMALITIES AND NEWLY DIAGNOSED PATIENTS WITH TYPE 2 DIABETES MELLITUS; A CASE-CONTROL STUDY.

CONTEXT: Nesfatin-1 is a novel peptide with both central and peripheral anorexigenic regulatory properties. Besides its effects on food intake, few studies have suggested a possible role for this peptide in the pathogenesis of diabetes mellitus type 2. OBJECTIVE: To compare serum levels of nesfatin-1 between healthy, normal-weight persons and three groups including healthy underweight, healthy obese and diabetic subjects. DESIGN: Prospective, case-control study, performed between January 2015 and January 2016. SUBJECTS AND METHODS: Fasting levels in serum nesfatin-1 were measured in 30 healthy, normal-weight individuals (controls), 30 healthy underweight persons, 30 healthy obese persons, and 30 patients with newly diagnosed diabetes type 2 using standard enzyme-linked immunosorbent assay (ELISA) kits. RESULTS: The mean serum nesfatin-1 level was significantly higher in controls (2.61 ng/mL) compared to that in obese (1.13 ng/mL) and diabetic (0.99 ng/mL) patients; and significantly lower than that in the underweight group (3.50 ng/mL). The obese and diabetic groups were comparable in this regard. No significant association was found between serum nesfatin-1 level and age, sex, or body mass index. CONCLUSIONS: Serum nesfatin-1 is possibly associated with weight-related abnormalities in otherwise healthy subjects and diabetes type 2. Obesity and diabetes type 2 may share a common pathologic point in this regard.

Infectious Causes of Bovine Abortion in Qazvin Province, Iran.

Abortion in dairy cattle is generally defined as the loss of fetus on days 47-265 of pregnancy and can significantly reduce the income and capital gain of farmers and producers. Therefore, sufficient economical support and preventive measurements are essential for this issue. In this study, genetic materials were extracted from the abomasum rennet, as well as homogenous preparations of brain, kidney, spleen, liver, and lung tissues of 128 aborted fetuses. Afterward, polymerase chain reaction (PCR) and Reverse Transcription Polymerase Chain Reaction (RT-PCR) tests were performed to identify the bovine viral diarrhea virus (BVDV), bovine herpesvirus 1 (BHV-1), Neospora caninum parasite, and serovars of Leptospira species. In the PCR test, the samples with 380 bp, 340 bp, and 173 bp bands were considered as positive for serovars of Leptospira, Neospora caninum, and BHV-1, respectively. Moreover, a 290 bp band was regarded to be BVDV in the RT-PCR. According to the findings of the current study, 39 (30.47%) of the samples were infected with Neospora caninum, 26 (20.31%) with BVDV, 17 (13.28%) with BHV-1, and 18 (14.06%) with serovars of Leptospira. In addition, multiple-agent infections were also detected in the samples. As a conclusion, the microorganism Neospora caninum was identified as the most prevalent infectious cause of abortion in the eight agro-industrial and livestock complexes, in Qazvin province, Iran.

Chitosan based hydrogels: characteristics and pharmaceutical applications.

Hydrogel scaffolds serve as semi synthetic or synthetic extra cellular matrix to provide an amenable environment for cellular adherence and cellular remodeling in three dimensional structures mimicking that of natural cellular environment. Additionally, hydrogels have the capacity to carry small molecule drugs and/or proteins, growth factors and other necessary components for cell growth and differentiation. In the context of drug delivery, hydrogels can be utilized to localize drugs, increase drugs concentration at the site of action and consequently reduce off-targeted side effects. The current review aims to describe and classify hydrogels and their methods of production. The main highlight is chitosan-based hydrogels as biocompatible and medically relevant hydrogels for drug delivery.

The effects of technical and compositional variables on the size and release profile of bovine serum albumin from PLGA based particulate systems.

Double emulsion solvent evaporation technique is one of the most attractive methods used to prepare micro and nanoparticles in pharmaceutical areas of interest, but because of the effects of many formulation factors on the size and release behavior of the fabricated particles, optimization of the formulation factors is needed. In this study various parameters including technical and compositional variables were considered to achieve an optimized formulation with desire characteristics especially size and the release profiles, using high shear homogenizer. In this regard, bovine serum albumin (BSA) was used as the model protein and double emulsion was formed with the addition of Tween 80 and Span 80 as surfactants for inner aqueous phase and oil phase, respectively. Hydroxypropyl beta cyclodextrin was used as protein stabilizer. After optimization steps, composite nanoparticles (core-shell) were made based on optimized formulation by hyaluronic acid as shell and poly lactic-co-glycolic acid (PLGA) as core material. Formulation of the BSA loaded PLGA nanoparticles using core shell strategy improved the release pattern of the BSA and diminished burst release. The final composite nanoparticles had the particle size of about 160 nm and 70 % of the loaded BSA was released during 14 days and the release data was better fitted to zero order release kinetics.

In vitro antibacterial evaluation of sol-gel-derived Zn-, Ag-, and (Zn + Ag)-doped hydroxyapatite coatings against methicillin-resistant Staphylococcus aureus.

Hydroxyapatite (HAp) coatings were applied using sol-gel method. Phosphor pentoxide and calcium nitrate were used as phosphorous and calcium precursors, respectively. Zinc nitrate and silver nitrate were used as substitute of calcium in HAp structure. As a base concentration, 1.5 wt %Ag and 2.5 wt %Zn were used. The weight percent of Ag was increased at 0.3 wt% and Zn content was scaled down at 0.5 wt%. Phase analysis and chemical bonds of synthesized materials were studied by XRD and FTIR. Antibacterial activity of Ag- and Zn-doped samples against methicilin-resistant Staphylococcus aureus (MRSA) were assessed by the plate-counting method. The XRD and FTIR results proved formation of HAp compound. Colony counting showed that silver and zinc ions prevent proliferation and growth of MRSA. Interestingly, co-presence of metal ions improves the antibacterial effectiveness of the coatings and the combined effect was greater than sum of the individual effects when each was administered alone. Overall, synergism between antibacterial activities of Zn(2+) and Ag(+) ions against MRSA can be suggested. Thus, cell toxicity decreases and biocompatibility increases without any decrement in antibacterial activity.

Preparation and assessment of chitosan-coated superparamagnetic Fe3O4 nanoparticles for controlled delivery of methotrexate.

In this study, Fe3O4 superparamagnetic nanoparticles were synthesized and stabilized by chitosan. Then the nanoparticles were characterized by Fourier transform infrared spectroscopy and transmission electron microscopy (TEM). Particle size distribution and Zeta potential of the particles also was assessed using Malvern Zetasizer. The paramagnetic behaviors of the uncoated and chitosan coated nanoparticles were measured using vibrating scanning magnetometry Particles morphology and size ranges of uncoated iron oxide nanoparticles were evaluated by TEM, showing uniform and narrow size distribution about 10 nm. After coating nanoparticles with chitosan and loading of methotrexate (MTX), the change in size was assessed using Zetasizer. Considerable increase in size was observed following the coating of the particles with chitosan and loading with MTX (the average size was 152 nm). Paramagnetic properties of the uncoated and chitosan-coated particles were assessed showing significant decrease in paramagnetic behavior after coating with chitosan, but it was enough to respond to the magnetic field. Finally loading efficiency, release rate and cytotoxicity of MTX were assessed indicating slow release behavior with the same levels of cell toxicity in SK-BR-3 cell lines, suggesting this formulation as a good candidate for the controlled delivery of MTX.

Reliability and validity of the Self-report Family Content Scale.

The aim of the study was to assess the reliability and validity of Self-report Family Content Scale for measurement of a contextual and psychoeducational model for family studies. The sample included 450 college students at Shiraz University (241 women, 209 men; M age=27.5 yr., SD=3.8). Principal-components factor analysis indicated a 7-factor solution. The mean correlations among factors and between factors and the total score of the scale were .18 and .42, respectively. Coefficients alpha indicated good reliability and internal consistency for the scale.

Preparation and in vitro evaluation of carrier erythrocytes for RES-targeted delivery of interferon-alpha 2b.

Carrier erythrocytes is one of the most promising systemic drug delivery systems investigated in recent decades. In this study, human erythrocytes have been loaded with interferon-alpha 2b (IFN) with the aim to benefit the reticuloendothelial system (RES) targeting potential of the carrier cells. Hypotonic preswelling method was used for drug loading in erythrocytes and the entire loading procedure was evaluated and validated. The loaded amount, entrapment efficiency and cell recovery of the loading procedure were 2906.33+/-588.35IU/0.1ml, 14.53+/-2.94%, and 83.61+/-0.49%, respectively, all being practically feasible. The carrier erythrocytes were characterized in vitro in terms of their drug release kinetics, hematological indices, particle size distribution, SEM analysis, and osmotic and turbulence fragility. IFN release from carrier cells was a relatively rapid process in comparison to the cell lysis kinetics, which is unusual considering the whole body of data published on this delivery system and other protein drugs, so far. All the tested in vitro characteristics showed significant, sometimes notable changes upon drug loading procedure, both with and without the drug.

Important criteria for spouse selection in a sample of Iranian youth.

The aim was to study important criteria for spouse selection by a sample of Iranian youth, 110 Iranian university students (60 women and 50 men; M age = 21.7 yr., SD = 1.5). Participants were single. Face-to-face interviews indicated that the order of the first 10 important criteria for the women were chastity, virtuous, religion, education, family characteristics, financial resources, job, commitment, social skills, and social prestige; for the men these were physical appearance, chastity, education, family characteristics, virtuous, religion, personality traits, social skills, housekeeping, and commitment and financial resources.

Preparation and in vitro characterization of carrier erythrocytes for vaccine delivery.

Erythrocytes as the most readily available and abundant cells within the body, have been studied extensively for their potential application as drug delivery carriers. In this study, human erythrocytes have been loaded by bovine serum albumin (BSA) as a model antigen/protein using hypotonic preswelling method for targeted delivery of this antigen to antigen-presenting cells (APCs). A series of in vitro tests have been carried out to characterize the carrier cells in vitro, including loading parameters, BSA and hemoglobin release kinetics, hematological indices, particle size distribution, SEM analysis, osmotic and turbulence fragility, and osmotic competency. BSA was loaded in erythrocytes with a loaded amount of 1.98+/-0.009mg with antigen release from carrier cells showing a zero-order kinetic consistent to that of the cell lysis. The apparent cell sizes, measured using laser scattering, were not significantly different from normal erythrocytes, but the real sizes, measured using SEM, and surface topologies were quite different between loaded and unloaded cells. The BSA-loaded cells were remarkably more fragile and less deformable compared to the normal cells. Totally, BSA-loaded erythrocytes seem to be a promising delivery system for reticuloendothelial system (RES) targeting of the antigens.

A postulated role of testosterone for prevention of cisplatin gonadal toxicity.

Testicular cancer is the most common solid tumor in young men. It has become one of the most curable solid neoplasms by applying multidisciplinary treatment approaches and new chemotherapeutic drugs. Cisplatin based chemotherapy as the most efficient chemotherapy of germ cell tumors has severe deleterious effects on all stages of spermatogenesis by various direct and indirect mechanisms. By marked improvement in oncologic control, prognosis and survival of patients with testicular cancer, their fertility, as one of the essential aspects of quality of life, is a matter of great concern. Since the probability and severity of spermatogenesis impairment is quite unpredictable during a course of cisplatin based chemotherapy, protecting the spermatogenesis during that phase by administration of exogenous testosterone in order to reduce the proliferation rate of germ cells would seem to be beneficial.

Antihirsutism activity of Fennel (fruits of Foeniculum vulgare) extract. A double-blind placebo controlled study.

Idiopathic hirsutism is defined as the occurrence of excessive male pattern hair growth in women who have a normal ovulatory menstrual cycle and normal levels of serum androgens. It may be a disorder of peripheral androgen metabolism. In this study we evaluated the clinical response of idiopathic hirsutism to topical Fennel extract. Fennel, Foeniculum vulgare, is a plant, which has been used as an estrogenic agent. The ethanolic extract of Fennel was obtained by using a soxhlete apparatus. In a double blind study, 38 patients were treated with creams containing 1%, 2% of Fennel extract and placebo. Hair diameter was measured and rate of growth was considered. The efficacy of treatment with the cream containing 2% Fennel is better than the cream containing 1% Fennel and these two were more potent than placebo. The mean values of hair diameter reduction was 7.8%, 18.3% and -0.5% for patients receiving the creams containing 1%, 2% and 0% (placebo) respectively.

Comparison of different gutta-percha root filling techniques: Thermafil, Quick-fill, System B, and lateral condensation.

OBJECTIVES: In this in vitro study, the core (gutta-percha or gutta-percha and carrier)/sealer ratio and the sealing ability were calculated for 4 different gutta-percha techniques: Thermafil, JS Quick-Fill, System B, and lateral condensation. STUDY DESIGN: One hundred single-rooted teeth were instrumented and obturated. Forty of the teeth were embedded in resin and sectioned horizontally at 1, 2,3, and 4 mm from the anatomic apex. Photographs were taken of each section, and the total area of the canal filled with core material or sealer was calculated. Sixty of the teeth were kept in wet storage for 90 days, after which the teeth were immersed in 2% methylene blue for 48 hours. The length of dye penetration was measured. RESULTS: Roots obturated with Thermafil and JS Quick-Fill contained significantly more core material than did those obturated with System B or with lateral condensation. The lateral condensation technique had the lowest core/sealer ratio (P < .05). Roots using Thermafil and Quick-Fill showed significantly less dye leakage than did the lateral condensation technique. CONCLUSIONS: Through use of these techniques in vitro, Thermafil and JS Quick-Fill with carrier and System B were found to be superior to the lateral condensation technique in terms of core/sealer ratio. Thermafil and Quick-Fill were superior to lateral condensation in terms of dye leakage.

An approach to controlled-release dosage form of propranolol hydrochloride.

It is possible to release a drug with only limited diffusion from a membrane-coated system using osmotic pumping. In this study, a propranolol osmotic pump was produced by coating the core tablets with cellulose acetate. The effects of membrane thickness, pore size, and stirring rate on the release rate of propranolol hydrochloride were studied. It was found that the thickness of cellulose acetate membrane had a profound effect on the release rate of propranolol hydrochloride from the membrane-coated tablets. The results showed that, when the membrane thickness increased, the release rate of propranolol decreased. The drug release follows a zero-order release when the delivery orifice is between 200 and 800 microns, but when the delivery orifice size is increased to 1000 microns, the release kinetic is abnormal. Fluid dynamics have an important effect on the delivery rate of propranolol from this device; the delivery rate increases as a function of the fluid flow. The drug release is higher under a turbulent condition with high rate of stirring.

Effect of polysorbates on atenolol release from film-coated tablets.

The effects of different concentrations of various polysorbates on the release rate of atenolol from film-coated tablets were evaluated. The release profile of atenolol showed that increasing the concentration of polysorbate resulted in an increase in the release rate of atenolol. The type of polysorbate had less effect on the release rate of atenolol. This study revealed that the release kinetic of atenolol from these film-coated tablets was a function of polysorbate concentration. Correlation coefficients of kinetic models could not solely determine the suitability of the models; the sum of the least square of differences also should be calculated when different kinetic models have similar correlation coefficients.

Evaluation of sealing properties of Thermafil and Ultrafil techniques in the absence or presence of smear layer.

The apical sealing abilities of two thermoplasticized gutta-percha techniques were evaluated. A total of 128 single-rooted teeth were divided into six groups in which Ultrafil, Thermafil, and lateral condensation techniques, with and without the smear layer, were used to obturate 20 roots in each group. The roots were implanted subcutaneously in rats for 90 days to expose the materials to vital tissue and tissue fluids. The roots were recovered, immersed in 1% methylene blue for 2 wk, cleared, and the dye penetration was measured using a dissecting microscope. Thermafil showed the least leakage with smear layer present and Ultrafil the least with it absent. The two thermoplasticized gutta-percha techniques both showed significantly less leakage than lateral condensation with and without the smear layer. Removing the smear layer reduced the leakage significantly in all groups.

Dentinal wall adaptation of thermoplasticized gutta-percha in the absence or presence of smear layer: a scanning electron microscopic study.

The dentinal adaptation of injected thermoplasticized gutta-percha and thermoplasticized gutta-percha resulting from Ultrafil and Thermafil systems was evaluated by scanning electron microscopy and compared with adaptation obtained with the lateral condensation of gutta-percha. Each method was evaluated with and without the smear layer removed. Scanning electron microscopic observations showed the thermoplasticized gutta-percha resulting from either system to have better dentinal wall adaptation than lateral condensation of gutta-percha in either the absence or presence of the smear layer. In the absence of smear layer, the adaptation of gutta-percha was improved in all groups. Although sealer was used, removing the smear layer was found to enhance the ability of the thermoplasticized gutta-percha to enter the patent tubules.

Provisional restorations for traumatically injured teeth requiring endodontic treatment.

The acid-etch-composite resin technique is used to retain a provisional restoration during the course of endodontic treatment. As described, the use of a gutta percha cone during the buildup and bonding to adjacent teeth provides the following: (1) good esthetics during endodontic treatment, (2) good support for rubber dam placement, (3) good entry to the canal, (4) good coronal seal for medication within the canal, and (5) good function as required by the provisional crown.