فهرست مطالب فهیمه نعمتی منصور

Recently, it has been found that tyrosol and farnesol can replace antibiotics due to their known antimicrobial and medicinal effects. Niosomes have been extensively researched for drug delivery during the last few decades, and their efficiency has been proven. The current study aimed to investigate the physical properties and antibacterial effects of niosomes loaded with tyrosol and farnesol.

Nanoniosomes loaded with farnesol and tyrosol were synthesized by the thin-layer hydration method. The physical properties of nanoformulations were measured using dynamic light scattering and scanning electron microscopy (SEM), and the release of farnesol and tyrosol from the nanocarrier was examined using a dialysis bag. In addition, Fourier-transform infrared spectroscopy was used to check the functional groups, and the stability studies of nanoniosomes were performed at temperatures of 25 °C and 4 °C for two months. Finally, its antimicrobial properties against the bacterial pathogens Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), and Pseudomonas aeruginosa (P. aeruginosa) underwent investigation. The cytotoxicity of free and farnesol- and tyrosol-loaded nanoniosomes on human foreskin fibroblast cells was also evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide method.

The size of nanoniosomes and farnesol- and tyrosol-loaded nanoniosomes was estimated to be 208 nm and 240 nm, respectively. SEM results indicated the spherical structure of niosomes. The slow release of farnesol and tyrosol from niosomes was observed, so that almost half of the drug was released from nanoniosomes after 72 hours. Nanoniosomes loaded with the above-mentioned drugs demonstrated good stability at 4 °C for 60 days. The results of the minimum inhibitory and bactericidal concentrations confirmed the antibacterial effects of both free farnesol and tyrosol and nanoniosomes loaded with farnesol and tyrosol against all three bacterial species, although these effects were more considerable for the nanoniosomes formulation.

Nanoniosomes loaded with farnesol and tyrosol have the potential to be used in the treatment of some common bacterial wound infections. However, clinical studies are needed in this field.

Farnesol- and tyrosol-loaded nanoniosomes had strong inhibitory effects on S. aureus, E. coli, and P. aeruginosa.

Proteolytic enzymes are essential for the survival of living organisms by irreversible hydrolysis of peptide bond. Carboxypeptidases are proteolytic enzymes found in many organisms and are widely used in various industries, such as food and pharmaceuticals. Aims of this study are expression of carboxypeptidase of chohnella A01, purification and determination of its activity.

The gene of carboxypeptidase of cohnella A01 has been cloned in pET26b(+) vector. Competent cells were prepared from E. coli BL21, plasmids were extracted and transformed to expression host. E. coli BL21 was cultured in LB medium and protein expression was induced by IPTG. After that cell harvested and recombinant protein was purified by Ni-TNA resin.

The expressed recombinant enzyme is a glutamate carboxypeptidase that capable of separating glutamate from folic acid.

Hence carboxypeptidase G has medicinal application and recombinant carboxypeptidase from cohnella A01 is a thermostable enzyme, after optimization, can be use it for medicinal application.

Chitosan is a biodegradable, biocompatible polymer regarded as safe for human dietary. Nanoparticles (NP) prepared with chitosan derivatives typically possess a positive surface charge and mucoadhesive properties such that can adhere to mucus membranes and release the drug payload in a sustained release manner. Current study is focused on preparation several kinds of chitosan formulations and different concentrations of cross linking agents in various conditions and characterization for achievement of most stable and biocompatible nanoparticle which labeled by FITC for in vitro tracking.
Analysis

In order to better investigate, in addition to chitosan, carboxymethyl chitosan was used. Preparation of chitosan nanoparticles is done by Ionotropic gelation method using different concentration of sodium tripolyphosphate (TPP) as cross-linking agent. Moreover, the synthesis of FITC-labeled chitosan at a combination of time, pH, different concentrations of salt and enzyme was investigated to produce stable nanoparticles for intestinal drug delivery. Physical characterization of nanoparticles were estimated by DLS and TEM as well as conjugation and covalent links were confirmed by FITR.

The size of the nanoparticles by transmission electron microscopy (TEM) was about 100±50.Hydrodynamic diameter was measured by DLS around 150±50 nm. In FTIR, covalent conjugation to the chitosan nanoparticles with control samples was confirmed. UV spectroscopy was used to determine the efficiency of conjugation which was over 70%.

All of these findings favor the notion that the FITC conjugated chitosan nanoparticles produced in our work may offer promise for the development of an efficient therapeutic and diagnosis carrier for colon cancer in the future. However, further investigation is required to provide more evidence on different aspects of the targeting activity and stability of this nanoparticle in in vivo.

Chlamydia trachomatis is one of the most common sexually transmitted diseases worldwide. Screening women for Chlamydia trachomatis in developing countries is highly desirable due to the ineffectiveness of the available diagnostic methods. The aim of this study is to design a detection kit for Chlamydia trachomatis in symptomatic women.

A total of 50 clinical specimens of symptomatic women were collected. The OMP1 gene sequence was extracted from the gene bank and the primer and probe were designed appropriately by Mega6 software. The extracted DNA was amplified by PCR. In order to provide positive control, PCR product was cloned into Pjet1.2 vector. To determine the sensitivity, DNA dilution was performed. To determine the specificity, DNA of vaginal bacteria was extracted and Real Time PCR was done.

In this study, we designed a kits with 100% specificity and 10pg/ϥl sensitivity in detecting chlamydial infection in symptomatic women.

With the use of designed primers and real time, we can detect 100% of chlamydia infections.

Background and purpose: Telomerase activity has a major role in acute promyelocytic leukemia (APL). It also has a critical role in disease recurrence. This research aimed at studying the cytotoxic effects of telomerase inhibition using oligonucleotide-based molecule against human telomerase RNA template (hTERC antisense) and non-nucleoside small molecule targeting catalytic subunit (BIBR5132) on APL-derived cell line. To evaluate whether inhibition of telomerase using either hTERC antisense or BIBR5132 could exert cytotoxic effect in APL, NB4 cells were subjected to different concentrations of the inhibitors and subsequent cell viability, metabolic activity, induction of apoptosis were investigated using Trypan blue assay, MTT, and annexin/PI staining, respectively. Also, Caspase-3 enzymatic activity and transcriptional alteration of apoptosis-related target genes were investigated. We found that targeting telomerase using hTERC antisense (45 pmol/L) and BIBR1532 (75 µL) for 48 h reduced the survival rate of NB4 cells nearly by 30% and 40%, respectively and induced a caspase-3-dependent apoptosis. Our results also suggest that suppression of c-Myc and subsequent increment of Bax/Bcl-2 ratio coupled with decreased telomerase activity may be rational mechanisms for the cytotoxicity of both telomerase inhibitors against NB4 cells. Current results clearly indicated that both BIBR1532 and hTERC antisense had anti-tumor activity against NB4 cells and anti-telomerase-based therapy may be an efficient treatment for acute promyelocytic leukemia.

Hepatitis is a systemic diseasethat causes liver inflammation. The prevention of this infection is a vaccination. The commonly used vaccine to fight this disease is to use the vaccine formulated with Alum. This vaccine cannot provide immune response and complete productivity in some people. In this study, cellular and humoral immune responses of hepatitis B vaccine were compared with hepatitis B vaccine formulated in MF59 adjuvant. In this experimental study, Balb/c mice received different formulations of the vaccine subcutaneously three times with a two-week interval. Then, the mice were bled and the levels of anti-HBs Ag were determined by the ELISA method. IFN-γ, IL-4, IL-2 and IFN-γ / IL-4 cytokines were examined by the ELISA method from the soup of spleen cells culture. The data were analyzed using the GraphPad prism software ANOVA. IL-4 levels were significantly higher in alum vaccine than the vaccine formulated in MF59, also the IFN-γ cytokine level showed no significant difference between two main groups. TNF-α cytokine shows that alum vaccine is more secreted due to the high inflammation compared with the vaccine with MF59. Total antibody in the third injection, in some dilutions of the commercial vaccine was more than vaccine with MF59. Significant decrease in IL-4 and antibodies indicates that the tendency of vaccine formulated in MF59 to induce cellular immune responses is higher than humoral immune responses. In addition, the safety and lack of side effects of the MF59 adjuvant can also be considered as another advantage.

Antibiotic resistance has prepared the way for substituting new therapeutic methods. Studies have indicated that nanoformulated antimicrobial agents have better therapeutic effects. In this study, the antimicrobial activity of chitosan nanoparticles loaded with amoxicillin (ACNs), was assessed in comparison with free amoxicillin against some Gram-positive and Gram-negative bacteria.
In this experimental study, the nanoparticles were prepared using the ionotropic gelation technique. The resulting nanoparticles were characterized by dynamic light scattering (DLS), scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. The antibacterial activity of amoxicillin and nanoparticles against standard and clinical strains of methicillin-susceptible and methicillin-resistant Staphylococcus aureus, Escherichia coli, and Enterococcus faecalis, was investigated by determination of minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and inhibition zone diameters. Data were analyzed using one-way ANOVA with Tukeyʼs post hoc test.
The size of chitosan nanoparticles (CNs) and ACNs was 88 and 106 nm, respectively. ACNs showed higher antibacterial activity compared to amoxicillin and CNs without drug as determined by the smaller MIC (0.375mg/ml) and MBC (2 mg/ml) values and larger zones of inhibition (28mm). The highest and the lowest sensitivity to nanoparticles, were observed for Enterococcus feacalis and methicillin-resistant Staphylococcus aureus, respectively.
The results of this study showed that the nanoformulation of amoxicillin may be an appropriate strategy to increase its therapeutic effects. However, precise clinical studies are required for its confirmation

Nanotechnology offers a great chance to treat drug-resistant microbial infections. The purpose of this study was to synthesize amoxicillin-encapsulated polymeric nanoparticles and compare the antibacterial activity of this nanoformulation with pure amoxicillin.
Amoxicillin-encapsulated polymeric nanoparticles were prepared using chitosan as a polymer and alginate as cross linking agent. The obtained nanoparticles were characterized by Fourier transform infrared, dynamic light scattering and scanning electron microscopy. The antibacterial effects of the nanoparticles were evaluated using broth microdilution and well diffusion methods against some common bacterial strains involved in hospital-acquired infections.
The drug-encapsulated nanoparticles were found to be spherical in shape with average size of 96 nm. These nanoparticles had a significant antibacterial effect on all tested bacteria, except for Pseudomonas aeruginosa. They also displayed stronger antibacterial activity than the nanocarrier alone or free antibiotic. The highest mean zones of growth inhibition (23.7 mm) for methicillin-sensitive Staphylococcus aureus and the smallest zone (12.5 mm) for the resistant species of this bacterium were determined. MIC of the nanoformulation against these two strains was respectively determined at 1.5 and 48 µg/ml and for Escherichia coli and Pseudomonas aeruginosa at 6.6 and 256 µg/ml.
The results suggest that amoxicillin encapsulation in polymer nanoparticles has the potential to increase its antibacterial activity against bacteria causing nosocomial infections.

Staphylococcus aureus is one of the major causes of nosocomial infections throughout the world. Efflux pumps such as norA play a major role in the development of resistance to different antibiotics in this bacteria. The aim of this study was evaluation of the antibiotic resistance and detection of efflux pump (norA) in methicillin resistant (MRSA) and ciprofloxacin resistant S. aureus isolates using genotypic and phenotypic methods.
During this sectional study, 250 clinical samples were collected from different hospitals in Tehran, Iran. S. aureus isolates were identified and the antimicrobial susceptibility patterns were determined. Ciprofloxacin minimum inhibitory concentration (MIC) was determined in both MRSA and ciprofloxacin resistant isolates. Furthermore, the presence of norA efflux pump gene in MRSA and ciprofloxacin resistant isolates was assessed phenotypically using ciprofloxacin and ethidium bromide MIC, with CCCP as efflux pump inhibitor, and genetically using PCR method.
Totally, 50 S. aureus isolates were recovered. The results of antibiotic susceptibility tests showed that 34 isolates (68%) were resistant to methicillin, of which 12 isolates (24%) were resistant to ciprofloxacin, as well. Moreover, all the MRSA- ciprofloxacin resistant strains harbored the norA gene and active efflux pump.
The results showed the correlation between ciprofloxacin resistance and norA efflux pump gene in MRSA isolates. Development of efflux pump inhibitors can be useful in the control of MRSA ciprofloxacin resistant strains.