Nova Biologica Reperta

A study on the targeting of ceramide metabolism by (-)-epicatechin gallate, catechin and quercetin in A-549 lung cancer cell line

Masoud Mashhadi Akbar Boojar

Catechin, epicatechin gallate (ECG) and quercetin, as bioactive flavonoids, have been shown to possess anticarcinogenic effects. Ceramide plays an important role in killing tumor cells. Accordingly, the aim of this study was to clarify the involvement of these compounds in ceramide metabolism in A549 cancerous cell line. Spectrophotometer, cell culture and HPLC methods were used. Cell viability index showed different potential of cytotoxicity effect for each of the studied agents, among which ECG was more potent. This index decreased significantly over 100 to 250 µM concentrations of treatment with respect to control. Cell treatments also caused considerable increase in ceramide level within cells in a dose-dependent manner. Sphingomyelinase activity increased significantly in treatment with quercetin and catechin. There was significant inhibition in acid ceramidase activity of cell extract in response to each of the three compounds, particularly over 100 µM in comparison with control. Data also showed no significant variation in glycosyl ceramide synthase activity in treated cells with quercetin, whereas the activity decreased significantly by Catechin and/or ECG. It is our conviction that different effects on ceramide metabolism enzymes may be related to various chemical groups on the common structure of the studied compounds. Due to structure-function relationship, these compounds had different effects on ceramide generation. Elevation in ceramide content in A549 cancer cell line induced apoptosis, which led to anti-cancerous effects, as observed in this study.

Comparison of three different staining methods for the morphometric characterization of Alpaca (Vicugna pacos) sperm, using ISAS® CASA-Morph system

Hernán Cucho

Sperm morphometry is a part of the semen analysis based on CASA technology and has shown a big role in the prediction of male fertility. This analysis implies the use of stain techniques, although it has been shown that staining can make dramatic changes in the cell morphometry in different species. The aim of the present work was to evaluate the differences in sperm morphometry in Alpaca, introduced by the use of different stain techniques. Single ejaculates from five adult alpacas were used. Samples were recovered by deferent duct deviation surgery technique. Three stain techniques (i.e. Hemacolor, Harri’s Henatoxylin and Diff-Quik) were evaluated. Morphometric analysis was done using ISAS® v1 CASA-Morph system, at 100x bright field objective magnification and a digital video camera with a final resolution of 0.08 µm/pixel. Almost 200 randomly selected cells were automatically analysed per sample and stain technique, considering several sperm-head and midpiece parameters regarding size and shape. Almost all of the parameters showed different levels of difference among the employed techniques. In general, the largest cell heads were the ones stained with Harri’s Hematoxylin and the smallest ones were the ones stained with Diff-Quik. The discriminant parameters used to differentiate between animals better were the head width, area and acrosome percentage. In conclusion, like in other species, alpaca sperm morphometry results were found to be sensitive to the applied stain technique. This implies the necessity of referring clearly the stain technique used in each case to perform comparisons between different works on the same species. None of the employed techniques was superior to the others. In addition, the method used to obtain the samples showed its usefulness and simplicity for repeated samplings.

Isolation and identification of alkaline protease-producing bacterium in the presence of washing-powder from Geinarje hot spring, Ardabil, Iran

Mahnaz Nasre Taheri

Proteases are important industrial enzymes used in different areas of industry, mainly detergent, food and leather industries. In this study, novel alkaline protease-producing bacterium was isolated from Geinarje hot spring and examined for maximum protease activity to be utilized in washing-powder. The isolated bacterium was cultured in mineral salt medium including 2% Skim Milk. Proteolytic activity of supernatant was measured by caseinolytic method. The effects of pH, temperature, SDS, Tween 80 and EDTA on protease stability and activity were investigated. The detergent compatibility of protease was assayed. On the basis of phylogenetic analysis and morphological as well as biochemical tests, the isolate was identified as a new strain of Brevibacillus borstelensis capable of generating extracellular alkaline protease. The generated protease was determined as alkaline metallo-protease having high activity at 60 ^oC and pH 9. Moreover, the alkaline protease was stable in the presence of SDS, Tween 80 and H₂O₂. It is compatible with commercial detergents. Finding proteases capable of degrading proteins in extreme environment (i.e. alkaline pH, high temperature and presence of surfactants) is valuable in biotechnological and industrial practices. Therefore, it can be utilized in detergent formulation in the future.

Induced genetic and morphological changes in Catharanthus roseus L. by cold atmospheric plasma

Zahra Noormohammadi

Catharanthus roseus is widely cultivated around the world. Cold atmosphere plasma (CAP) has been used to improve crop plants seed germination, and plant growth. In the present study, three different times of CAP (40, 50 and 60s) were used for the treatment of C. roseus seeds, and the changes in morphological traits, antioxidant enzymes and the genetic content of the treated plants were examined. Cold plasma (50s) markedly raised the seed germination, length of the leaves and the stem length of the plants in comparison with those in the control group. The catalase and peroxidase enzyme activities had the highest value in 60 and 40 s CAP treated plants, respectively. The sequence related amplified polymorphism (SRAP) markers showed the highest degree of genetic diversity in 50s cold plasma treated plants (Ne = 1.388, I = 0.316, He = 0.217, uHe = 0.237 and P % = 50.08). Based on Neighbor Joining, principle coordination analysis (PCoA) and analysis of molecular variance (AMOVA) test, four significantly distant groups were formed. The 40s and 50s cold plasma treated plants stand far from the control plants due to genetic difference. The results indicate that cold atmosphere plasma could be used as an economic and environmentally safe tool in increasing C. roseus growth characteristics in addition to inducing genetic variations.

Polyploidy Induction in German Chamomile (Matricaria chamomilla L.) by Herbicide Trifluralin

Leila Karami

Polyploidy induction in German chamomile (Matricaria chamomilla L.) by herbicide trifluralin

Leila Karami¹, Mohammad Modarresi², Mohammad Amin Kohanmoo², Fatemeh Zahabi Ahmadi² & Saeed Irian³
¹Department of Horticulture, Faculty of Agriculture and Natural Resources, Persian Gulf University, Bushehr, 7516913817, Iran; ²Department of Plant Breeding, Faculty of Agriculture and Natural Resources, Persian Gulf University, Bushehr, 7516913817, Iran;³Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
Correspondent author: Leila Karami, leila.karami@pgu.ac.ir

Abstract. Chamomile (Matricaria chamomilla L.) is a medicinal herb belonging to Asteraceae family. This study was conducted to investigate the effect of trifluralin on the induction of polyploidy in chamomile. Two independent full factorial experiments on seedlings and 2-leaf stage apical buds were performed. Following morphological, biochemical and cytogenetic analysis, chromosome numbers of 18 and 36 were detected for diploid and tetraploid types, respectively. Morphological and biochemical examinations revealed that an increase in the number of full chromosome set results in a reduction in stomata number per unit area as well as an increase in stomata size, chloroplast number, and chlorophyll content. Induction of ploidy level increments also reduced plant height and increased the number of lateral branches, leaf size, and diameters of stems, flowers and receptacles. It is concluded that a concentration of 22.5 µM trifluralin in both methods is optimum for the production of tetraploid chamomile with the highest rate of polyploidy induction and the lowest percentage of abnormality.

The distribution patterns of Astragalus sect. Hypoglottidei DC. (Fabaceae) in Iran

Mohammad Mahmoodi

The distribution patterns of taxa belonging to the section Hypoglottidei DC. were investigated using DIVA-GIS software. Based on Zohary classification, five global patterns, with the exception of widely distributed species, include 1. Centro-Asiatic province, 2. Irano-Anatolian province, 3. Pontic province, 4. East Mediterranean subregion and 5. West Mediterranean subregion, were identified. In addition, the Iranian species of this section were classified in three patterns: 1. Central Zagros 2. Khorasan-Kopet Dagh 3. Central and Eastern Alborz. The maps and distribution patterns of the species were also presented.

Stipa richteriana (Poaceae) and Galium songaricum (Rubiaceae): two new additions of the Central Asian species to the flora of Iran

Farshid Memariani

Stipa richteriana and Galium songaricum are newly recorded species for the flora of Iran, collected from Binalood and Hezar-Masjed Mountains in Razavi Khorassan Province. The geographical distribution of both species is mainly confined to the Middle Asia. Morphological characters of two newly recorded species are compared with their close relatives. Notes on taxonomy, ecology, phytogeography, and conservation status of both species are provided.

Scorzonera incisa (Asteraceae), as a new record from NW Iran

Seyed Reza Safavi

Scorzonera incisa, known as an endemic plant to Turkey, has been collected from West Azarbayejan Province, NW. Iran. The species is reported for the first time from the country; it is very similar to Scorzonera calyculata, which is distributed in the western half of Iran and east of Iraq. However, with regard to the particular shape of the achenes of Scorzonera incisa, this species could be separated from Scorzonera incisa. In this paper, the description, the image of the herbarium sample and the distribution map of this species are presented.

Nutlet micromorphology of the genus Marrubium L. and allies and its systematic implication (Lamiaceae: tribe Marrubieae)

Yasaman Salmaki

Nutlets of 20 taxa of the tribe Marrubieae were examined by scanning electron microscopy (SEM) and detailed descriptions of nutlet micromorphological features for all examined taxa are provided. The nutlets exhibited variation in size, shape, color and surface sculpturing. The nutlets shape of most species studied is ovate, but rounded, broad ovate, elliptic, lanceolate, triangular and oblong can also be found in a few species. Six basic types of the sculpturing pattern of nutlet surface can be distinguished: reticulate, foveolate, scalariform, ruminate, pusticulate and colliculate-granulate. The most common type of nutlet sculpturing among the studied species is reticulate, but the variation in size and shape of their composing cells provided useful diagnostic characters. Our investigation revealed that the type of sculpturing was more useful in taxon delimitation among Marrubium species and allies at the species rank.

A new record of Lepidium (Brassicaceae) for the flora of Iran

Yasamin Nasseh

In this paper, the new taxa Lepidium ferganense (Brassicaceae) was recorded for the flora of Iran for the first time. This taxon belongs to Irano-Turanian region. It was compared with the closest taxa in the present study. In addition, a morphological description, several Illustrations, and a distribution map were presented.

Taxonomic significance of achene morphology in the genus Centaurea L. (Asteraceae)

Farideh Attar

Achene morphology of 49 taxa of the genus Centaurea L. was studied in terms of 19 different characteristics. On the basis of the variation in these features, some sections, such as sect. Cyanus with hairy hilum, were separated. Despite various differences, C. leuzeoides and C. gilanica were categorized in the section Psephelloideae, a section with lots of character variations within its species. Also, C. albonitens has different characteristics in comparison with other taxa within section Phaeopappus, for example features such as pappus color and the ratio of pappus length to that of achene separated the species from other members of the section. The last but not the least, highly different characteristics were observed in the achene morphology of C. phlomoides as compared with other species in section Cynaroides. Differences such as short pappus and hilum length were found to separate C. phlomoides from other members of sect. Cynaroides. In conclusion, on the basis of the findings of this research, most achene morphological characteristics can be appropriately used as key features for the differentiation of sections in the genus Centaurea. An identification key based on the features of the achene, images related to the achene of the studied species were also presented