Nova Biologica Reperta

Electrochemical assay of anti-tetanus toxoid monoclonal antibody by silver enhancement of gold nanoparticles at carbon nanotubes modified glassy carbon electrode

Ahmad Molaei Rad

Tetanus is caused by the toxin secreted by Clostridium tetani. Due to the rapid infection with this bacterium, it is so important to investigate the tetanus immunity of people. Therefore, electrochemical biosensors, as one of the most effective tools in this regard, have demanded characteristics such as being fast, simple, cost-effective and portable. However, their detection sensitivity is not sufficient. Hereon, silver enhancement of gold-nanoparticles was proposed for the improvement of detection. Hence, the current study applied gold-nanoparticles as label, following with silver enhancement, to investigate the yes/no electrochemical detection of anti-tetanus toxoid antibodies in the indirect immunoassay utilizing glassy-carbon electrodes modified by carbon nanotubes. The analytical procedure consists of the reactions of the tetanus toxoid with the antibody at electrode, so that followed by the interaction of gold-labeled secondary antibody and then silver enhancement process. In this study, the cyclic-voltammeter variation and difference of gold to silver signal based on silver ions fluctuations were also investigated. The results indicated that ∆E_p increased from 0.24 V before silver enhancement reaction to 0.57 V after the silver enhancement. The results also demonstrated that after silver enhancement, current significantly increased and current plot at E^c_p transferred to positive potentials and at E^a_p moved to negative potentials. In conclusion, this method increases the detection sensitivity and can simply use to other bio-molecules detection.

The effect of a VEGFB antagonist peptide on the expression level of miR-210 in a mouse model of breast cancer

Soheila Talesh Sasani

Breast cancer is the fourth common cancer worldwide and occurs when breast cells begin to uncontrolled division and tumor formation. Angiogenesis is one of the essential factors in cell growth and maintenance of homeostasis in the natural and pathological conditions, while VEGFs are the most critical factors in angiogenesis. MiR-210 plays an important role in the angiogenesis via association with VEGF. Here, the miR-210 expression changes in response to a VEGFB antagonist peptide, called VEGB1, was studied in female BALB/c mice bearing 4T1 cell line induced breast tumor. The treated group received 1mg.kg^-1and 10mg.kg^-1 of the peptide and the control group received PBS intraperitoneally during two weeks. Both of the animal groups underwent a resection of breast tissue 14 days after treatment and miR-210 expression level was investigated. Statistical analysis by On-way ANOVA showed that the expression level of miR-210 gene had significant differences among the groups treated with various doses of VEGB1. Also, the gene expression was significantly different between peptide-treated groups and control samples (p<0.05). MiR-210 expression level had 42% reduction in mice treated with 1mg.kg^-1of VEGB1, while 90% was seen in mice treated with 10mg.kg^-1 of VEGB1 showing the inhibitory function of VEGB1 antagonist peptide at different doses.

Evaluation of antioxidant and anticancer effects of Lavandula angustifolia and Melissa officinalis on HeLa, OVCAR-3 and MCF-7 cancer cell lines

Parichehr Hanachi

From ancient times, plants have been regarded as therapeutic agents, in addition to their usage as food. Plants are rich sources of antioxidant and phenolic compounds. Lavandula angustifolia and Melissa officinalis are medicinal herbs rich in antioxidant compounds. The aim of this study was to compare the antioxidant and anticancer properties of Lavandula angustifolia and Melissa officinalis extracts using aqueous, ethanol and methanol solvents, to select the best extraction methods and solvents and to evaluate the cytotoxic effect of the extracts on HeLa, OVCAR-3 and MCF-7 cancer cell lines. Methanol, aqueous and ethanol extracts were obtained from the dried leaves of the plants and the antioxidant activities of each extract were measured by DPPH and FRAP methods. Finally, the anticancer effects of the extracts on HeLa, OVCAR-3 and MCF-7 cancer cell lines were evaluated by MTT assay in order to identify the most efficient extract. Comparing the results of total antioxidant assay showed that the highest amount belonged to the ethanol extract of Melissa officinalis and Lavandula angustifolia methanol extracts using lyophilization method. The IC₅₀ value of ethanol extract of Melissa officinalis was equal to 0.028 mg/ml on OVCAR-3 cells, which was the best result obtained in comparison with other solvents, and the ethanol extract of Lavandula angustifolia with IC₅₀ = 2.07 mg/ml on MCF-7 cells was the most effective extract among the others. In HeLa cell-line, methanol extract of Lavandula angustifolia with IC₅₀= 7.36 mg/ml showed the highest cytotoxicity. In this study, for the first time, the effects of different extracts of Lavandula angustifolia and Melissa officinalis on MCF-7, OVCAR-3 and HeLa cancer cells were evaluated and the results showed that ethanol and methanol extracts of these plants had better toxic effect on cancer cells.

The effect of exosomes derived from human ovarian epithelial cancer cells on the secretion of AMH and Inhibin in granulosa cells

Javad Baharara

Exosomes are secreted by different types of cells and known as biological packages. Exosomes have significant role in intercellular communications and involved in the development and progression of various diseases such as cancer. Inhibin B and anti-mullerian hormone (AMH) are markers of granulosa cell tumors (GCT) and due to the role of exosomes in the progression of cancer, in this experimental study, the effect of exosomes derived from human ovarian cancer cells on the secretion of Inhibin B and antimullerian hormone (AMH) by granulosa cells was investigated. First, A2780 human ovarian cancer cells were cultured, then the supernatant was collected to extract the exosomes by ultracentrifugation and subsequently, the extracted exosomes were checked out using dynamic light scattering (DLS) and Scanning electron microscopy (SEM). In addition, granulosa cells were isolated and cultured from the ovaries immature female Balb / C mice and treated with 25 μg/ml of exosomes derived from the ovarian cancer cell-line. Inhibin B and AMH hormones levels were then measured. The results showed the significant (P<0/05) increase of the level of inhibin B and AMH hormones in the treated cells in comparison with the control group. According to the results, exosomes increased the secretion of Inhibin B and AMH hormones and seems to be effective in the proliferation of granulosa cell tumors.

Prevalence and intensity of coccidian blood parasite infection in three species of lizards from Markazi Province, Iran

Hossein Javanbakht

Coccidian (Apicomplexa: Coccidia) parasites have been reported in almost all groups of vertebrates. These parasites may cause serious ecological and pathological effects to their hosts. Reptiles were found the be the host of the parasites of the genera Schellackia and Lankesterella, as these parasites undergo their entire life cycle in the reptilian body, with an intermediate stage of dormancy in the tissues of a hematophagus transmitter (generally a mite or a mosquito), until the next reptilian host swallows the infected insect. However, very little is known about the prevalence and intensity of these parasites in most of their distribution area, which is the focus of the current study. To do so, the prevalence and intensity of these blood parasites in three species of lizards, including Ophisops elegans, Trapelus lessonae and Eremias monticola from Markazi Province of Iran were investigated. Samples of blood were collected from the caudal vein of 51 lizards of the aforementioned species by insulin syringes, then the blood smears were dried, fixed with methanol and stained with Giemsa solution. Prepared blood samples were then examined by means of light microscopy, using a 100× magnification objective lens. 41 percent of the 51 examined samples were found to be infected by haemococcidians blood parasites from the family Lankesterellidae. The mean intensity of parasites in the three species were between 0.03-0.05%. The investigation revealed a relatively high percentage of prevalence and intensity of blood parasites in lizards of central part of Iran. The results showed the presence of a complete set of various vectors for the spread of haemoparasites in the area. Therefore, future researches are strongly recommended to utilize genetic markers in order to obtain a better identification of the parasites.

Sexual dimorphism in digit length ratios of the second to fourth digits (2D:4D) in Blanford's fringe-toed lizard Acanthodactylus blanfordi Boulenger, 1918 (Sauria: Lacertidae) in the south of Iran

Nastaran Heidari

Finger length ratios are organized during embryonic development of fingers as they exposed to sex steroid hormones, and may show varying degrees of sexual dimorphism between males and females in different animal groups. Among all the finger length ratios calculated in a sample, the ratio between the second to fourth fingers (2D: 4D) is the most important one. In this study, the 2D:4D ratios in both sides of the body (right and left) in all limbs were investigated to determine if sexual dimorphism is present in the 2D: 4D ratios in 44 specimens studied (20 males and 24 females) of Acanthodactylus blanfordi. Other morphological traits of the two sexes were also examined (23 metric and meristic traits), as a result, sexual dimorphism was observed in five metric and meristic morphological traits. In terms of the ratio of the size of the fingers and toes, sexual dimorphism in the length of the fingers was observed only in 2D: 4D on the right side of the body in forelimbs and hindlimbs. The value of this trait was higher in males than females and this difference was statistically significant P ≤0.05).

Molecular phylogeny and taxonomy of species of Cousinia section Stenocephalae (Asteraceae) in NE of Iran

Shahrokh Kazempour-Osaloo

In this study, the phylogeny and morphological variation of 6 species of the genus Cousinia sect. Stenocephalae, i.e., C. hypopolia, C. decipiens, C. esfandiarii, C. renominata, C. stahliana and C. daryoushiana, distributed in NE regions of Iran, were investigated. ITS, ETS and rpl32-trnL ^(UAG) sequences for 12 species (17 specimens) were generated. Phylogenetic interspecies relationships were constructed by Bayesian method and likelihood analysis. The specimens were also investigated in terms of morphological characters. Despite very similar morphological characters and geographical distributions, the separation of these species was confirmed by the obtained results of the molecular data (combined data of plastid and nuclear sequences). On the basis of the present study, the above-mentioned species were almost separated from each other by morphological characters and molecular data. A diagnostic key and the description of these species were also presented. These results did not conform with the previous palynological and anatomical studies of the species.

The floristic study of Golestankooh area in Isfahan Province, Iran

Azadeh Akhavan Roofigar

Golestankooh, with an area of 130000 ha, is located in Isfahan Province. The present study was carried out in order to identification of flora of the aforementioned region, as a result, 620 taxa in total, belonging to 61 families and 303 genera, were identified. Asteraceae, represented by 104 species, was the largest family, following with Fabaceae, Lamiaceae and Brassicaceae with 63, 60 and 59 species, respectively. Astragalus, with 33 species, was the most diverse among the genera studied. Hemicryptophytes with 41% and therophytes with 34% are dominant life forms in the region. With regard to geographical distribution, 300 species (48%) were found to be Irano-Turanian elements. Moreover, 130 species, 21% of the identified plant taxa from the region, were found to be endemic to the flora of Iran. Due to high species diversity in the Golestankooh area, it seems necessary to use appropriate strategies for their conservation.