Nova Biologica Reperta

Co-expression of recombinant human nerve growth factor with trigger factor chaperone in E. coli

Zahra Hajihassan

Nerve growth factor (NGF) is a neurotrophic factor that is functional in the survival, maintenance and differentiation of nervous system cells. This protein has three subunits, of which the beta subunit has the main activity. Its structure consists of a cysteine knot motif made up of beta strands linked by disulfide bonds. It can be used as a therapeutic agent in the treatment of many diseases. As NGF extracted from natural sources is unsuitable for therapeutic goals, many studies have attempted to produce recombinant β-NGF. In this study, Trigger Factor (TF) chaperone was expressed simultaneously with β-NGF in E. coli in order to obtain increased yield of soluble recombinant human β-NGF. For this purpose, pET39b (+)::β-NGF and chaperone plasmid pTf16 were transferred to E.coli (DE3 strain). After the induction of each promoter, the total proteins and periplasmic proteins were extracted. To confirm the effects of TF on total protein and soluble β-NGF expression level, Bradford and Dot blot techniques and ImageJ software were used. Then, β-NGF was purified using affinity chromatography column (Ni⁺²-NTA). Also, the PC12 cells were treated with the protein for one week in order to study the function of purified NGF. Our data indicated that co-expression of TF could increase the soluble and periplasmic production of β-NGF but not total proteins. Also, the treatment of PC12 cell line with purified β-NGF, co-expressed with TF chaperone, showed differentiation of these cells to nerve cells. This indicated that the purified NGF is fully functional. Our data suggest that the co-expression of cytoplasmic chaperone (TF) with recombinant nerve growth factor might be an efficient approach to produce a proper quantity of soluble and active rhNGF.

Anti-cancer effects of the methanolic extract of Persian Gulf brittle star

Sajjad Farrokyar

Nowadays natural compounds are increasingly used in various fields since they cause fewer adverse side effects than their chemical counterparts. In addition, preparation and production of natural compounds are comparatively simple. The present study aimed to evaluate the anti-cancer effects of brittle star methanol extract on MCF-7 cells. Anti-cancer effects of brittle star methanol extract were examined on the MCF-7 cell line. Moreover, the toxicity of this compound was assessed using cell apoptosis methods, ethidium bromide/acridine orange (EB/AO) staining and Annexin V Apoptosis Detection Kit. According to the MTT assay, the methanolic extract of brittle star could prevent proliferation in a time- and dose-dependent manner. The calculated inhibitory concentration that decreased the cell-viability by half (IC₅₀) was estimated at 150 µg/ml. On the other hand, results of EB/AO staining were indicative of increased apoptotic cells in the treatment group. In addition, results of Annexin V/PI protocol demonstrated a significant enhancement in the percentage of late apoptotic cells (29.02%) in the treatment groups (150 µg/ml) and the percentage of necrotic cells were 2.77% for a period of 24 hours. According to the results, the methanolic extract of Persian Gulf brittle star induced apoptosis in the MCF-7 cell line. These effects could be attributed to the presence of certain active biological compounds, such as Saponins, Naphthoquinones and Carotenoids. Furthermore, our findings suggested that brittle star might have beneficial effects on cancer management. Therefore, this natural marine compound could be used as a complementary element in the prevention and treatment of cancer.

Frequency of two VKORC1 gene variants and its correlation with warfarin maintenance dose

Mohammad Khalaj-Kondori

Warfarin is a commonly-prescribed anticoagulant used to treat and prevent thromboembolic events. The requirement for varying doses of warfarin depends on genetic and environmental components. In this study, the frequency of two single-nucleotide polymorphic variants of the vitamin K epoxide reductase complex subunit 1 (VKORC1) gene (1173 C>T (rs9934438) and 3730 G>A (rs7294)) and its correlation with warfarin maintenance doses were investigated in patients with heart valve replacement from West Azarbayejan, Iran. Blood samples were obtained from 185 patients; their genomic DNA was extracted and samples were genotyped by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) assay. To assess if the blood warfarin level is different among genotypes, we used a one-way analysis of variance (ANOVA) followed by a Tukey’s post-hoc comparison. The minor allele frequency was determined to be 54% for 1173T and 53.7% for 3730A. Patients who carried the G allele at position 3730 and T allele at position 1173 required a significantly lower daily mean warfarin dosage (P <0.001). Consideration of the VKORC1 gene polymorphism, especially at the initial stages of the therapy, can be helpful in pre-treatment dosing of warfarin, which, in turn, reduces the adverse effects resulting from inappropriate drug prescription.

Morphological, anatomical and palynological studies of the genus Zoegea L. (Asteraceae) in Iran

Khadijeh Mahmoodi

The genus Zoegea L. belongs to Asteraceae family and has about 10 species in the world. This genus is considered to be an Irano-Turanian and Mediterranean element and is distributed in south-western and central Asia and in the central, southern, north-western and south-western parts of Iran as well. The subspecies classification of the genus is not consensus and various classifications could be found in different taxonomy resources. In this study various specimens from different regions of Iran were studied. In addition, anatomical and palynological characters were used to perform a cluster analysis in order to determine species groups. In the end, our results confirmed that Z. baldschuanica and Z. glabricaulis were distinct species.

New chromosome counts in some plant species from east and south-east Iran

Firouzeh Bordbar

In this study, we investigated the chromosome numbers of some endemic species distributed in East and South-East Iran. The mitotic chromosome counts were presented for the first time for six species in three angiosperm families including: Linaria iranica (2n=12), Nanorrhinum campyloceras (2n=18), Chaenorhinum grossecostatum (2n=24) (Plantaginaceae), Gaillonia bruguieri (Rubiaceae) (2n=22), Nepeta rivularis and N. assurgens (Lamiaceae) (2n=18).

Plant-based expression systems for protein and antimicrobial peptide production

Azadeh Niknejad

Molecular farming technology offers a unique advantage that almost any protein can be produced economically and safely under very controlled conditions. Besides traditional production systems, such as bacteria, yeasts, insects and mammal cell lines, plants can now be used to produce eukaryotic recombinant proteins, especially therapeutic ones. Their advantages as hosts for protein production include correct post-translational modifications, low-cost maintenance and no risk of contamination by human pathogens. The system is widely applied in agriculture and industry, especially in life science and pharmaceutical industry. The application of transgenic plants in the production of vaccines, antibodies and pharmaceutical proteins has been playing a key role in plant genetic engineering in recent years. The production of recombinant proteins plays a critical role in the production of high amounts of high-quality proteins. In this review, common problems in the production of recombinant proteins and antimicrobial peptides in plant-based expression systems are discussed and strategies for their solution are suggested. Viral vector-mediated transient gene expression in plants enables rapid production of pharmaceutical proteins such as vaccine antigens and antibodies. To conclude, plant-based systems have the potential to bring unique efficacy-enhancing features to increase the utility and effectiveness of vaccines and therapeutics.

Floristic study of vegetation in Palang Galoun protected region, Isfahan province, Iran

Fateme Sadeghipour

The Palang Galoun protected region, comprising 34935 hectares, is located 75 km northwest of Najaf Abad, and 102 km northwest of Isfahan city. The aim of this research was to assess the floristic spectrum, life form varieties, chorological analysis, determination of protected status and identification of medicinal, threatened species criteria and poisonous plants of this natural heritage. Plant samples were collected during different vegetative seasons at several stages. Life forms of samples were determined and chorological analysis was performed. On the basis of the obtained results, a total of 166 species belonging to 126 genera and 39 families were identified. Six families, 23 genera and 26 species belonged to monocotyledons, whereas 33 families, 103 genera and 140 species belonged to dicotyledons. In terms of chorological analysis, 58% of plant species were distributed in the Irano-Turanian region. It is noteworthy that 44 endemic species, 97 medicinal species, 48 pasture species and 23 poisonous species were present in the studied area. Life forms were found to comprise 54% hemicryptophytes, 24% therophytes, 10% geophytes, 7% chamaephytes and 5% phanerophytes. Based on IUCN red list categories and criteria, there were 22 species in the lower risk category and seven species categorized as vulnerable case.

The genus Allium (Amaryllidaceae) in Iran: on the status of Allium ampeloprasum L. and its relatives

Shahryar Saeidi Mehrvarz

Allium ampeloprasum (Amaryllidaceae) is recorded as a new taxon for the flora of Iran. It is morphologically most similar to Allium atroviolaceum and Allium iranicum. This species is compared with its two aforementioned relative species and some notes are given on its affinities. A full description, images and a distribution map are also provided. In addition, a lectotype is selected for A. atroviolaceum.

Cousinia elymaitica (Cardueae, Asteraceae) a new species from west of Iran

Farideh Attar

Cousinia elymaitica, which has been collected from western Iran, is described in this paper as a new species. This new species has been assigned to the sect. Pugioniferae on the basis of its habit, purple corolla and decurrent leaves. Its closest related species is C. macroptera. A distribution map and an illustration of the related specimens have also been provided.

Two new species and a new record of the genus Astragalus (Fabaceae) from NE Iran

Yasamin Nasseh

During the study on the specimens of Astragalus sect. Ammodendron in the herbarium of Ferdowsi University of Mashhad (FUMH), based on the last and most recent sources, two new species, i.e. A. microfoliolatus sp. nova and A. rashed-mohasseli sp. nova were found and described. Morphological evidence confirmed that the belonging of these taxa to Astragalus sect. Ammodendron. The most important differences between A. microfoliolatus and its closest relatives are leaflets with very small size and glabrescent legume. Also, the main distinctive characters ofA. rashed-mohasseli are the presence of hairs on the dorsal side of vexillum, oblong legume and short rachis. Hairy vexillum has been obseverd for the first time in this section. In addition, A. aiwadzhi is recorded as a new species for the flora of Iran. Taxonomic descriptions, illustrations and distribution maps are provided to expedite identification.

Plantago lagocephala (Plantaginaceae), a new record for the flora of Iran

Shahryar Saeidi Mehrvarz

Plantago lagocephala is recorded as a new species for the flora of Iran from Razavi Khorassan Province. It is compared with the closely related species including P. lachnantha, P. ciliata and P. psammophila. Moreover, the diagnostic morphological characters and a distribution map are provided. The conservation status of the new record is evaluated based on the IUCN red list categories and criteria.

Lepidium virginicum L. (Brassicaceae), a new record for the flora of Iran

Alireza Naghinejad

In the framework of floristic and vegetation studies of different parts of the Hyrcanian region of northern Iran, Lepidium virginicum L. (Brassicaceae) is reported here for the first time for the flora of Iran. Along with a distribution map for the species, an identification key is provided for the nine annual or biennial species of Lepidium, already reported from Iran.

Gypsophila hispida, A New record for flora of Iran

Atiye Nejad Falatoury

Gypsophila hispida, sect. Hispidae, is reported for the first time from NW Iran. Illustrations, morphological and palynological descriptions and distribution are provided in detail, along with a key for the six taxa of subgen. Hagenia from Iran.

Novelties in Cardueae (Asteraceae)

Mansour Mirtadzadini

The current investigations suggest the segregation of Jurinea cartilaginea from Jurinea, and its placement among Centaureinae. Molecular studies, based on the nuclear ribosomal internal transcribed spacers (nrITS), chromosome number data and pollen morphology, support the morphological results. Based on these results, J. cartilaginea is related to and, therefore, transferred to the genus Karvandarina. Due to the morphological characters differing from Karvanderina aphylla, a new subgenus, Pseudojurinea, is described. Molecular study also supports the monophyly of Jurinea.