Cloning, Expression of Hirudin Gene from Leech (Hirudo orientalis) in BL21(DE3) Strain

The use of leeches in bloodletting therapy is one of the most important techniques in ancient medicine from Greek time up to now. Leeches saliva contain anticoagulant hirudin which facilitate blood flow, this is one reason for using leeches to save thousands of severed fingers, noses and ears in recent years. In this study, hirudin gene successfully cloning and expressed in BL21(DE3) strain. Leech type was detected as H. orientalis by using two types of genes 18SrDNA and CO-I genes, Hirudin gene was amplified by specific primers from H. orientalis cDNA. Furthermore, Hirudin gene was expressed in BL21(DE3) strain under the control of T7 promoter in pET-16b vector, constructed vector pET-16-HR vector was extracted and used to amplify hirudin gene by specific primers, then hirudin gene band was appeared after digestion of extracted plasmid with Hind III and NdeI restriction enzyme. Hirudin expression was established by Real-time PCR. Production of hirudin established in LB medium and purified by IMAC column, DEAE Sepharose and SP Sepharose. Concentration of produced hirudin within its solution was measured by ELISA kit which reached to 1.35ng, thrombin titration method was used to determine hirudin activity which showed Hirudin protein required 360µl from thrombin for clot formation

Construction of Synthetic Intact Human Parathyroid Hormone Gene and Testing the Transformation Efficiency and Expression of it in E. coli strains

 human parathyroid hormone gene (hPTH gene) is responsible for preproparathyroid hormone production (biologically inactive hormone) which further modified to form the biologically active hormone, a synthetic hPTH gene with 289bp which cod for active intact hPTH was constructed by Five long primers by splicing overlap extension PCR (SOE), the gene contain a de generated codon, and have two recognition site of restriction enzymes in both ends of the Gene, and a stop codon. The synthetic hPTH gene was transformed in two E. coli strains DH5? and BL21 (DE3) with transformation efficiency6.6* 106 and 3.4* 106 for both isolatesrespectively. And the transformation of gene was detected by extraction of constructed plasmid and amplification of cloning gene on it, the expression of the cloning gene in the BL21 (DE3) was detected by performing Real Time PCR which gave a Ct value of about (23.46). That makes the synthetic gene suitable for direct expression of human active protein inside the bacterial cells

Biotechnology and the developing world

The life sciences offer opportunities for revolutionizing human welfare activities. Enriched by inputs from genomic research, biotechnology is a major force for development in all countries. Entwined with culture and socio-ethical values, biotechnology contributes to solving problems like food and water insecurity that impede national development and threaten peace in the developing world. The lack of facilities and professional skills in biotechnology limits R & D initiatives in the developing and the least developed countries (LDCs); and, restricts their full participation in take-off activities in national and self-reliant regional ventures in sustainable development. The practice of biotechnology different in many developing countries is nevertheless impressive. The establishment of biotechnology parks and medicinal plant farms in several developing countries is indicative of biotechnology being accorded high policy status in national development; of its significance in the eradication of poverty; and of its use in the empowerment of women in applying the technology for human and social welfare. This review provides several examples of different types of biotech activities that are being employed for development in the developing world

First report of grapevine dieback caused by Lasiodiplodia theobromae and Neoscytalidium dimidiatum in Basrah, Southern Iraq

In Basrah, grapevines suffer from dieback. Lasiodiplodia theobromae and Neoscytalidium dimidiatum were isolated from diseased grapevines, Vitis vinifera L. and identified based on morphological characteristics and DNA sequence data of the rDNA internal transcribed spacer (ITS) region. The results of the pathogenicity test conducted under greenhouse conditions for L. theobromae and N. dimidiatum reveal that both species were the causal agents of grapevines diebacks in Basrah, Southern Iraq. A brief description is provided for the isolated species.Key words: grapevine, dieback, Lasiodiplodia theobromae, Neoscytalidium dimidiatum, internal transcribed spacer (ITS), rDNA, Iraq

Marjoram Relaxes Rat Thoracic Aorta Via a PI3-K/eNOS/cGMP Pathway.

Despite pharmacotherapeutic advances, cardiovascular disease (CVD) remains the primary cause of global mortality. Alternative approaches, such as herbal medicine, continue to be sought to reduce this burden. is recognized for many medicinal values, yet its vasculoprotective effects remain poorly investigated. Here, we subjected rat thoracic aortae to increasing doses of an ethanolic extract of (OME). OME induced relaxation in a dose-dependent manner in endothelium-intact rings. This relaxation was significantly blunted in denuded rings. N(ω)-nitro-l-arginine methyl ester (L-NAME) or 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ) significantly reduced the OME-induced vasorelaxation. Cyclic guanosine monophosphate (cGMP) levels were also increased by OME. Moreover, wortmannin or LY294002 significantly reduced OME-induced vasorelaxation. Blockers of ATP-sensitive or Ca2+-activated potassium channels such as glibenclamide or tetraethylamonium (TEA), respectively, did not significantly affect OME-induced relaxation. Similarly, verapamil, a Ca channel blocker, indomethacin, a non-selective cyclooxygenase inhibitor, and pyrilamine, a H1 histamine receptor blocker, did not significantly modulate the observed relaxation. Taken together, our results show that OME induces vasorelaxation via an endothelium-dependent mechanism involving the phosphoinositide 3-kinase (PI3-K)/ endothelial nitric oxide (NO) synthase (eNOS)/cGMP pathway. Our findings further support the medicinal value of marjoram and provide a basis for its beneficial intake. Although consuming marjoram may have an antihypertensive effect, further studies are needed to better determine its effects in different vascular beds

Ethnopharmacology and therapeutic potential of Anchusa strigosa: a comprehensive review

Anchusa strigosa Banks and Sol. is a rough flowering plant of the Boraginaceae family native to Eastern Mediterranean region that is widely used in traditional herbal medicine, mainly for the treatment of wounds, abdominal pain, and arthritis, to name a few. This article aims to gather knowledge related to the medicinal properties of A. strigosa. Specifically, it summarizes its traditional uses and pharmacological activities in the treatment of various diseases. Moreover, its botanical, ecological, and phytochemical characteristics are also discussed. Research showed that this plant is rich in pyrrolizidine alkaloids, particularly in the leaves. Other bioactive metabolites identified in this plant include flavonoids, phenolic acids, triterpenes, organic acids, and volatile organic compounds. These phytochemicals are responsible for the reported pharmacological properties of A. strigosa, including antimicrobial, antioxidant, anticancer, anti-inflammatory, antiarthritic, gastric protective, antidiabetic, and pro-wound healing. This warrants further investigation into the molecular mechanism of action behind the observed effects to elucidate its therapeutic potential. Nevertheless, more research on this plant is needed to ensure its efficacy and safety

Assessment of genetic diversity in Iraqi camel breeds using Cytochrome b

Joudi breed were collected from Basrah, Al-Muthanna, Najaf, Babylon, and Wasit (10 samples each). As well as 15 samples of the Khawar breed from Anbar and 15 samples of the Hurra breed from DhiQar. The analysis of molecular variance (AMOVA) was employed This study aimed to investigate the genetic diversity, population structure, and demographic history of Iraq camel breeds. Eighty blood samples were randomly collected from unrelated animals from different parts of Iraq. Fifty samples of the Joudi breed were collected from Basrah, Al-Muthanna, Najaf, Babylon, and Wasit (10 samples each). As well as 15 samples of the Khawar breed from Anbar and 15 samples of the Hurra breed from DhiQar. The analysis of molecular variance (AMOVA) was employed in order to determine the genetic variability within and among populations of this camel breeds. The neutrality tests and mismatch distribution analysis were also applied to assess the neutrality and demographic expansion of the populations, respectively. The results revealed a total of 16 different haplotypes with high haplotype diversity (0.648) and low-nucleotide diversity (0.00109). The haplotypes of the Cytb gene were 15 haplotypes, 10 of which were in the Joudi breed, 6 in the Hurra breed and one haplotype in the Khwar breed. The haplotype H-2 was common to the three breeds. The variations within and among populations accounted for 94.36 and 5.64% of the total variation, respectively. The results of the neutrality test for Cytb showed that the Joudi and Hurra breeds had negative values for both Tajima's D and Fu's Fs. The highest values were -1.00737 and -1.98591 for the Hurra breed and the lowest values were for the Joudi breed (-2.14737 and -6.59079 respectively). In the Khwar breed, the values were zero for Tajima's D and Fu's Fs and the obtained result conforms to the model of population expansion (t > 0 and θ1> θ0) for Joudi and Hurra breeds

The Therapeutic Wound Healing Bioactivities of Various Medicinal Plants

The skin serves as the body’s first line of defense, guarding against mechanical, chemical, and thermal damage to the interior organs. It includes a highly developed immune response that serves as a barrier against pathogenic infections. Wound healing is a dynamic process underpinned by numerous cellular activities, including homeostasis, inflammation, proliferation, and remodeling, that require proper harmonious integration to effectively repair the damaged tissue. Following cutaneous damage, microorganisms can quickly enter the tissues beneath the skin, which can result in chronic wounds and fatal infections. Natural phytomedicines that possess considerable pharmacological properties have been widely and effectively employed forwound treatment and infection prevention. Since ancient times, phytotherapy has been able to efficiently treat cutaneous wounds, reduce the onset of infections, and minimize the usage of antibiotics that cause critical antibiotic resistance. There are a remarkable number of wound-healing botanicals that have been widely used in the Northern Hemisphere, including Achiella millefolium, Aloe vera, Althaea officinalis, Calendula officinalis, Matricaria chamomilla, Curcuma longa, Eucalyptus, Jojoba, plantain, pine, green tea, pomegranate, and Inula. This review addresses the most often used medicinal plants from the Northern Hemisphere that facilitate the treatment of wounds, and also suggests viable natural alternatives that can be used in the field of wound care

Ziziphus nummularia attenuates the malignant phenotype of human pancreatic cancer cells: Role of ros

Pancreatic cancer (PC) is the fourth leading cause of all cancer-related deaths. Despite major improvements in treating PC, low survival rate remains a major challenge, indicating the need for alternative approaches, including herbal medicine. Among medicinal plants is Ziziphus nummu-laria (family Rhamnaceae), which is a thorny shrub rich in bioactive molecules. Leaves of Ziziphus nummularia have been used to treat many pathological conditions, including cancer. However, their effects on human PC are still unknown. Here, we show that the treatment of human pancreatic ductal adenocarcinoma cells (Capan-2) with Ziziphus nummularia ethanolic extract (ZNE) (100–300 µg/mL) attenuated cell proliferation in a time-and concentration-dependent manner. Pretreatment with N-acetylcysteine, an ROS scavenger, attenuated the anti-proliferative effect of ZNE. In addition, ZNE significantly decreased the migratory and invasive capacity of Capan-2 with a concomitant downregulation of integrin α2 and increased cell–cell aggregation. In addition, ZNE inhibited in ovo angiogenesis as well as reduced VEGF and nitric oxide levels. Furthermore, ZNE downregulated the ERK1/2 and NF-κB signaling pathways, which are known to drive tumorigenic and metastatic events. Taken together, our results suggest that ZNE can attenuate the malignant phenotype of Capan-2 by inhibiting hallmarks of PC. Our data also provide evidence for the potential anticancer effect of Ziziphus nummularia, which may represent a new resource of novel anticancer compounds, especially ones that can be utilized for the management of PC

Origanum syriacum L. Attenuates the Malignant Phenotype of MDA-MB231 Breast Cancer Cells

Breast cancer is the leading cause of cancer-related deaths among women. Among breast cancer types, triple negative breast cancer (TNBC) is the most aggressive, and is resistant to hormonal and chemotherapeutic treatments. As such, alternative approaches that may provide some benefit in fighting this debilitating pathology are critically needed; hence the utilization of herbal medicine. Origanum syriacum L., one of the most regularly consumed plants in the Mediterranean region, exhibits antiproliferative effect on several cancer cell lines. However, whether this herb modulates the malignant phenotype of TNBC remains poorly investigated. Here, we show that in MDA-MB-231, a TNBC cell line, Origanum syriacum L. aqueous extract (OSE) inhibited cellular viability, induced autophagy determined by the accumulation of lipidized LC3 II, and triggered apoptosis. We also show that OSE significantly promoted homotypic cell-cell adhesion while it decreased cellular migration, adhesion to fibronectin, and invasion of MDA-MB-231 cells. This was supported by decreased activity of focal adhesion kinase (FAK), reduced α2 integrin expression, and downregulation of secreted PgE2, MMP2 and MMP-9, in OSE-treated cells. Finally, we also show that OSE significantly inhibited angiogenesis and downregulated the level of nitric oxide (NO) production. Our findings demonstrate the ability of OSE to attenuate the malignant phenotype of the MDA-MB-231 cells, thus presenting Origanum syriacum L. as a promising potential source for therapeutic compounds for TNBC