Vol. 7 & Issue 3 ; Section B : Biological Sciences

Date : May 2017 to July 2017

 

Development of Techniques for Identification of Protein Composition in Mammal Sperm Fluid by Comparison with Other Biological Probes from Mammals, by Combining Different Assay Methods. A Pilot Study

*Iskra V Sainova, Ilina Valkova, Tzveta Markova, Elena Nikolova,Angel Baldzhiev, Angel Alishev, Marin Nenchev

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3; 701 -709

[DOI: https:/doi.org/10.24214/jcbps.B.7.3.70109]

  • Abstract

    Protein composition (types of proteins and eventual protein-protein interactions) in mammal sperm fluid as a biological material, for which is proposed to be supplemented with different molecules, responsible for the control in the life processes of the male germ cells, was isolated and investigated. The protein content was subsequently compared with that of other biological materials, containing previously low-differentiated normal adult stem/progenitor cell types, as well as with protein extracts from anatomic organs from adult mammals, containing normal cell types in different stages of maturation and differentiation. Combinations of various methods for assay were developed and tested. The strongest similarities in the mammal sperm fluid protein composition with this from biological probe, rich of different normal stem/progenitor cell types, were established. The developed methods for assay, in support of many literature findings, are usable about more precise identification of the composition of the biological structures, but also for identification proteins and protein-protein interactions. As one of the key molecules in these processes was characterized the tri-peptide Glutathione, in particular it’s reduced from (GSH). As a proof about that was accepted the established increased levels of this molecule in the process of myeloid cell differentiation. The label-free LC-MS/MS assay was characterized as a usable technique, possessing many advantages in comparison with other proteomics technologies. The combined technique of laser irradiation of biological objects with their 3D-visualization gives a possibility about more precise identification of the changes, occurring in different phases of living processes, on both cellular and organism levels. Future investigations and development of other methods for identification of concrete biological molecules, as well as direct and/or indirect interactions between them, should be performed.

Melamine content in some milk-based products

Semaghiul Birghila, Mihaela Mirela Bratu, Maria Roman

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3; 710 -717

[DOI: https:/doi.org/10.24214/jcbps.B.7.3.71017]

  • Abstract

    This work reports the analysis of melamine in forty samples (22 of powdered milk and 18 chocolate) purchased from local market in Romania, in order to examine the presence of melamine over baseline level in products. The European Union has a clear legislation establishing a standard limit of the melamine content in milk powder for children. An analytical method including liquid extraction followed by mixed mode ion exchange/reversed phase solid phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used. The method is sensitive (limit of detection and quantification are 0.05 mg/kg and 0.150mg/kg), precise (RSDs<6.5%) and accurate, with recoveries averaged between 82.68 - 100.55%, for all fortification levels and matrices. Melamine was detected in 32 out of 40 samples, at concentrations ranging from 0.064 to 0.326 mg/kg. The low levels of melamine in analyzed samples indicated that these do not represent a health risk.

Antibacterial activity of Herbal and Spice Extracts on Bacterial Clinical Isolates of Urinary Tract Infections

Mir Naiman Ali and Mohammed Mazharuddin Khan

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3; 718-732,

[DOI: https:/doi.org/10.24214/jcbps.B.7.3.71832]

  • Abstract

    In the present study 370 urine samples were collected from two diagnostic laboratories of Hyderabad, Telangana State, India, of which 244 urine samples were tested as UTI positive and 116 bacterial cultures were isolated belonging to 5 species: Escherichia coli (48); Klebsiella pneumoniae (30); Pseudomonas aeruginosa (24); Enterococcus faecalis (8) and Proteus mirabilis (6). Three selected plants (Cinnamomum cassia, Syzygium aromaticum and Azadirachta indica) and their parts (bark, flower, and leaf) were used to test their antibacterial activity by agar well diffusion assay and MIC was determined. Three extracts- aqueous, ethanolic and methanolic were tested; highest antibacterial activity was observed with ethanolic extracts of Cinnamomum cassia with strong effect on E. coli and lowest against K. pneumoniae with diameter of inhibition zones (DIZ) of 20.56 ± 0.40 and 15.66 ± 0.57 mm respectively. Preliminary phytochemical analysis of the plant parts revealed the presence of active compounds such as flavonoids, phenolics, tannins and alkaloids. The results obtained in this study revealed broad spectrum antibacterial activity of selected plant extracts on all five UTI bacterial isolates compared with standard antibiotics used for UTI treatment.

Dietary Use of Ethno Gynaecological Plants by Tribal Women of Melghat

Suruchi Naik and Rekha Sharma

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3; 733-741.

[DOI: https://doi.org/10.24214/jcbps.B.7.3.73341]

  • Abstract

    The tribal people are indigenous people who depend on the forests and the traditional healers for their day-to-day and medicinal needs. Melghat is a tribal belt of Maharashtra where more than 80% population belongs to different tribes. The objective of the present investigation was to study the dietary use of ethno gynaecological plants and confirm their medicinal properties from traditional medicinal systems of Ayurveda viz., Indian Pharmacopeia and Dravyagunvidnyan. For the investigation, eight villages located on the boundary of Melghat Tiger Reserve were selected. The local medical practitioner’s viz., Bhagat, Bhumkas and Daais, of seven different tribes were interviewed. The medicinal properties of ethno medicinal plants were studied under four categories viz., menstrual problems like menorrhagia, hypo-menorrhea, leucorrhoea, menorrhea, Amenorrhea and menstrual pain; contraception, abortion and delivery; fertility and pregnancy; post pregnancy and lactation. About 29 medicinal plants were found to be used by tribal as ethno gynaecological medicines. The study revealed the resemblance between the traditional nutrition knowledge and Ayurveda database. Documentation of this knowledge may have significant importance to explore further concepts of ethno gynaecology and dietary studies.

Molecular cloning of the α-Gliadin genes from Thinopyrum and evolution analysis with those from
Triticeae

Qing-Rui Wang, Da-Zhi Wang, Huai-Yu Yang, Wen-Jia Wang, Xi-Han Ding,Shan-Shan Zhai, Li-Ping Bai, Zhi-Fu Guo

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3; 742-756.

[DOI: https://doi.org/10.24214/jcbps.B.7.3.74256]

  • Abstract

    As all seed storage proteins, gliadins are digested to provide nutrients for the embryo during seed germination and seedling growth stages. In this study, eight coding sequences of α- liadin genes designed as TpGlia1, TsGlia1, TcGlia1, TcGlia2, TcGlia3, TcGlia4, TjGlia1 and TjGlia2 were isolated from the genomic DNA of Thinopyrum, which encoded the mature proteins with 206, 276, 296, 307, 319, 279, 290 and 304 amino acid residues, respectively. Comparative analysis showed that all the isolated genes had typical structural characters of α-Gliadin genes that have been reported in previous. Particularly, apart from TpGlia1 and TcGlia3, the rest of other gliadins were recognized as pseudogenes due to the in-frame stop codons. The number of Cys in TpGlia1 and TcGlia 4 are 2 and 5, respectively, which play a key role in deciding the structure and function of protein. Phylogenetic analysis suggested that TcGlia 4 was closely related to those from V genome of Dasypyrum breviaristatum, and TcGlia1 was clustered into a group of α-gliadin genes from S or H genome of Elymus sibiricus, A genome of Triticum aestivum and B genome of Triticum aestivum; TpGlia1, TjGlia1 and TcGlia3 seemed to be more homologous with the gene sequences from Ee genome.

LC/PDA/ESI-MS/MS Polyphenols Profiling in the In vitro Active Leaves Extracts of Combratum hartmannianum against human Pathogens with Special Emphasis to Madurella mycetomatis

Hiba A. Ali, Abdelhalim A. Hamza, Ayman E. Abass, Omima E. Ahmed

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3; 757-774,

[DOI: https://doi.org/10.24214/jcbps.B.7.3.75774.]

  • Abstract

    The present communication represent an attempt to investigate the antimicrobial and antimycetomal activity of the leaves of Combratum hartmannianum (Combretaceae) and to define the phytochemical profiles of the active agents. Decoctions of the leaves of C hartmannianum are commonly used in Sudanese traditional medicines against jaundice, external skin infections, malaria and similar febrile diseases. Air dried ground leaves of C. hartmannianum were extracted using 80% methanol. The methanolic extract was sequentially fractionated with petroleum ether, chloroform and ethyl acetate. The aforementioned extracts of C. hartmannianum were tested against two Gram positive and three Gram negative bacteria as well as two fungi. Additionally, the obtained extracts of C. hartmannianum were tested in vitro against Madurella mycetomatis the most common eumycetoma causative organisms employing a newly developed microtitre plate- based antibacterial assay incorporating resazurin as an indicator of cell growth.Following bioactivity guided fractionation the ethyl acetate phase at both concentrations (1mg/ml, 5mg/ml) was significantly active against Staphylococcus aureus (20mm, 20mm) and Escherichia coli (20mm, 20mm). Furthermore, this fraction at a concentration of 5mg/ml possessed activity against activity against Protues vulgaris (17 mm), Pseudomona. aeruginosa,(20 mm) Aspergillus niger (20 mm) and Candida albicans (20 mm). The leaves chloroform extract (1mg, 5mg) possessed high activity against Bacillus subtitles (30mm, 25mm) and S. aureus (30mm, 25mm). the petroleum ether extracts of the leaves at two different concentrations (1mg, 5mg) showed activity against Staphylococcus aureus (30 mm) Bacillus subtitles (18mm, 23 mm) and Escherichia coli (25 mm). A promising inhibitory activity emerged against Madurella mycetomatis ranging between 78 and > 39.1 μg / ml. Most active were the ethyl acetate and chloroform fractions with MIC > 39.1 μg / ml. polyphenols were mainly accumulated in the chloroform and ethyl acetate phase which showed very similar TLC and HPLC chromatograms. Reverse phase High Pressure Liquid Chromatography coupled to Tanden Mass Spectromentry performed on the ethyl acetate fraction of the leaves of C. hartmannianum led to the identification of sixteen flavonoids and a phenantherene which were believed to be responsible of the activities mentioned above. These results support the different traditional uses associated with the plant studied. Pharmacological merits reported on C. hartmannianum were also in agreement with the results obtained.

The interaction between liver oxidoreductases in experimental thermal injury

Soloveva Anna and Didenko Natalya

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3, 775-783,

[DOI: https://doi.org/10.24214/jcbps.B.7.3.77583]

  • Abstract

    In this work the interaction between lactate dehydrogenase (LDH) and aldehyde dehydrogenase (ALDH) was studied in normal and thermal injury because thermal trauma is one of the central places in surgery and traumatology. The experiment was carried out on white Wistar rats. All animals under anesthesia (Zoletil (60 mg/kg) + XylaVET (6 mg/kg)) received flame burns on 20% of the skin surface. The catalytic and kinetic properties of aldehyde dehydrogenase and lactate dehydrogenase were determined in subcellular fractions of rat liver. In vitro we investigated the effect of Na lactate and Na pyruvate on the ALDH activity of intact rats and rats with thermal injury. It was established that during the burn the activity of ALDH and LDH in direct and reverse reactions decreased in the mitochondria. There was a high degree of positive correlation between the activities of LDH in direct reaction and ALDH (r=0,995; p< 0.0001) in thermal injury. In vitro it was revealed that pyruvate had a negative effect and lactate had positive effect on the activity of ALDH in the liver with intact rats and rats with burn.

Natural dye ultrasound extraction from beetroot: role of extraction solvent pH on color and enzyme inactivation

Blanca Edith Esquivel-González, Olga Miriam Rutiaga-Quiñones, Nuria Elizabeth Rocha-Guzmán, Luis Medina-Torres, Elizabeth del Carmen Varela- Santos, Luz Araceli Ochoa-Martínez

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3; 784-796,

[DOI: 10.24214/jcbps.B.7.3.78496]

  • Abstract

    The presence of enzymes in agricultural commodities causes quality changes, including color, flavor, and nutritional losses. The use of non-thermal and innovative technologies for the production of quality extracts derived from plant products is a recent trend in food processing. The aim of this study was to evaluate the effect of ultrasound applying different pH in the extraction solvent for obtaining a natural dye from beetroot on the color and enzyme inactivation. The natural beetroot dye was obtained using the extraction solvent (Mc Ilvaine buffer) at pH of 3.5, 5.0 and 6.5, ultrasound amplitude of 40, 60 and 80% and time of processing of 2, 6 and 10 minutes. Physicochemical parameters, color and enzymes were determined on the dye extracted, additionally, the microstructure of beetroot mash was analyzed. It was found that the physicochemical parameters were minimally modified when the pH of the solvent extraction was changed during ultrasound application. Concerning to the color characteristics, hue angle showed intense red color for ultrasound treatments at 3.5 pH. In the same way, this pH leads to obtaining the complete inactivation of peroxidase (100%) and 98.35% of inactivation for polyphenol oxidase for treatments at 10 min and 40% amplitude, which coincides with the treatment selected as optimal. From the microstructural analysis, it was possible to observe the formation of microchannels on the tissue surface beetroot as a result of ultrasonic waves. Therefore, ultrasound extraction at acidic pH could be employed for extracting a natural dye from beetroot in an effective way.

Could Sodium Benzoate Enhance Broad Bean Salinity Tolerance?
III. Antimicrobial Potency, Antioxidant Activity and Phytochemical Constituents

Bardees M. Mickky, Muhammad A. Abbas, Esraa D. Elmahdy and Raneem H. Youssef

JCBPS; Section B; May 2017-July 2017; Vol.-7; No.-3, 797-806.

[DOI: 10.24214/jcbps.B.7.3.79706]

  • Abstract

    The objective of the present study was to investigate the impact of seed presoaking in sodium benzoate (SB) on seawater (SW)- stressed broad bean (Vicia faba L.) at early germination stage. Methanolic extracts of 7- day old seedlings were prepared to assess their antimicrobial efficacy, antioxidant criteria and phytochemical profile. The results obtained indicated that the extracts of all treatments has no effect on Streptococcus pyogenes or Proteus vulgaris. Meanwhile, Bacillus subtilis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli and Candida albicans were greatly inhibited by the extracts especially those of SB- treated and/ or SW- stressed seedlings. It was recorded also that SB could enhance the ability of some extracts to scavenge diphenyl-picrylhydrazyl (DPPH) and H2O2 with accompanied increase in their reducing power and total antioxidant activity. Through phytochemical analysis, proteins, alkaloids, saponins and calcones were found to be absent from the extracts of all the considered treatments. However, reducing sugars, carbohydrates, amino acids, phenols, flavonoids, tannins, phloba-tannins, steroids, terpenoids and anthraquinones were all present but to various levels. Based on the resulted data, seed presoaking in SB was recommended to enhance the antimicrobial and antioxidant activity of SW- stressed broad bean seedlings

Variation of Chlorophyll Amount in Some Landscape Plants: a case study of Rize

Ilknur Zeren, Alican Cesur, Elnaji A. Ahmaida Saleh, Mansour Mosii Mohammed Mossi

JCBPS; Section B; May 2017 – July 2017, Vol. 7, No. 3; 807-817,

[DOI: 10.24214/jcbps.B.7.3.80717]

  • Abstract

    Chlorophyll, the pigment that which gives green color to plants, and Provides for the realization of the photosynthesis phenomenon in which the oxygen and nutrients are necessary for the life of all creatures. Studies have shown that the amount of chlorophyll in plants varies according to many factors. Plant type and growing conditions are at the top of these factors. In this study, it was aimed to determine the variation of chlorophyll content in some plant species which are used extensively in landscape studies in Rize city center. Rize has the highest annual rainfall in Turkey and the average annual rainfall is over 1400 mm. Therefore, the study was carried out in Rize city center and measurements were made in 16 plant types used in landscape studies in city center as 5 repetitions. Study results were evaluated by Variance Analysis and Duncan Test, lowest and highest chlorophyll values and standard deviations of species were determined. According to the results of the variance analysis, there was a significant difference in 99,9% confidence level among the species, chlorophyll levels of the tested species were collected in 9 homogeneous groups according to Duncan test results. The lowest amount of chlorophyll was found in Prunus cerasifera (17,40 cci) and the highest amount of chlorophyll was found in Citrus reticulata (129,04 cci).