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Yousef M Al-saraireh,1 Fatemah OFO Alshammari,2 Ahmed MM youssef,3 Sameeh Al-Sarayreh,4 Ghadeer H Almuhaisen,5 Nedal Alnawaiseh,6 Jehad M Al-Shuneigat,4 Hamzeh M Alrawashdeh7 1Department of Pharmacology, Faculty of Medicine, Mutah University, Al-Karak, Jordan; 2Department of Medical Laboratory Technology, Faculty of Health Sciences, The Public Authority for Applied Education and Training, Shuwaikh, Kuwait; 3Department of Pharmacology, Faculty of Pharmacy, Mutah University, Al-Karak, Jordan; 4Department of Biochemistry and Molecular Biology, Faculty of Medicine, Mutah University, Al-Karak, Jordan; 5Department of Microbiology and Pathology, Faculty of Medicine, Mutah University, Al- Karak, Jordan; 6Department of Public Health, Faculty of Medicine, Mutah University, Al-Karak, Jordan; 7Department of Ophthalmology, Sharif Eye Centers, Irbid, JordanCorrespondence: Yousef M Al-sarairehDepartment of Pharmacology, Faculty of Medicine, Mutah University, PO. Box: 7, Al-Karak, 61710, JordanTel +962799172658Email Yousef.sar@mutah.edu.joPurpose: Colon cancer is a leading cause of mortality worldwide. It has a relatively poor prognosis; therefore, new therapies are needed. One of the tumour-related enzymes that has gained considerable interest is CYP4Z1. This enzyme has been expressed in many tumours and has been hypothesized as a potential biomarker or target for novel anticancer therapies.Patients and Methods: CYP4Z1 overexpression was immunohistochemically examined in a large panel of colon tissue types including normal, benign, primary and metastatic ones, and the enzyme’s relation to histopathological features and patient survival was evaluated.Results: A high CYP4Z1 expression was observed in benign, primary and metastatic colon tissues compared to a weak or lack of expression in normal tissues. Importantly, there was a significant differential in CYP4Z1 expression where it was stronger in metastatic, primary and benign, respectively (p < 0.05). A significantly high rate of CYP4Z1 expression was found in high histological grades and late stages of the disease, where its expression was more evident in patients with metastasis in the lymph nodes (p < 0.05). Interestingly, CYP4Z1 expression was identified an independent prognostic predictor of poor overall survival of colon cancer patients (p = 0.003).Conclusion: CYP4Z1 was distinctly overexpressed in benign, primary and metastatic colon tissues compared to corresponding normal tissues. This differential in CYP4Z1 expression across different types of colon tissues strongly supports CYP4Z1 as potential biomarker and target for novel anticancer therapy development.Keywords: cancer, colon cancer, cytochrome P450, cytochrome 4Z1, immunohistochemistry

In this study, the regulation of ascorbate peroxidase (APX) specific activity, anthocyanin, carotenoid, hydrogen peroxide, lipid peroxidation, and protein levels in cress leaves in response to different abiotic stresses were investigated. The total APX specific activity was significantly elevated after 9 days of drought treatment, short-term (2 h) exposure to 10, 100 and 370 µE of light, long-term exposure (at least 6 days) to 100 mM NaCl versus the specific APX activity in the controls. Furthermore, a significant change in total APX activity was detected in response to treatment with different temperatures; this change was an early response to 4 °C and 30 °C for a maximum of 4 h, while short-term exposure to 35 °C did not change total APX activity. The results of the present study revealed that plants have a wide range of mechanisms to cope with different stresses that possibly involve morphological changes. The results indicated that Lepidium sativum plants launch common protective pathways only under drought, salinity and high light stresses, while other protective mechanisms/strategies could be responsible for increasing the plants tolerance towards temperature and low light. Future studies will investigate changes in the photosynthetic quantum yield and specific target metabolites, proteins, and nonenzymatic antioxidants.

Background: There is a high variability in chemical composition of essential oil from Artemisia herba-alba grown in different countries and different localities in the same country. This has led to the characterization of many oil-dependent chemotypes assigned to the plant. Only one report was published on the essential oil composition of Artemisia herba-alba grown in Jordan. Aim: The current study aims to determines the essential oil composition of Artemisia herba-alba grown wild in south Jordan and test their activity against clinical isolate antibiotics resistant bacteria. Methodology: The essential oils were isolated by hydrodistillation and analysed by Gas Chromatography-Mass Spectrometry (GC-MS). The screening for essential oil activity was carried out using disc diffusion method on methicillin-resistant Staphylococcus aureus , methicillin-sensitive

Faculty of Medicine, Department of Biochemistry and Molecular Biology, Mutah University, Mutah, Jordan. Faculty of Medicine, Department of Pharmacology, Mutah University, Mutah, Jordan. Faculty of Science, Department of Chemistry, Albalqa' Applied university, Albalqa, Al-Salt, Jordan. Faculty of Science, Department of Chemistry, Yarmouk University, Irbid, Jordan. Faculty of Medicine, Department of Microbiology and Immunology, Mutah University, Mutah, Jordan.

Background: A biofilm is group of bacteria embedded in a self-produced matrix made of polysaccharide and protein. Bacterial biofilms cause chronic infections because they show increased tolerance to antibiotics and disinfectant chemicals as well as resisting phagocytosis. Biofilm bacteria show much greater resistance to antibiotics than their free-living counterparts. Essential oils are a cheap and effective alternative to antibiotics that could be used to combat antibiotics resistant microorganisms. The aim of the present study was to determine the effects of essential oil of wild Thymus vulgaris grown in Jordan on biofilm-forming clinical bacterial isolates. Results: All of the isolates included in this study were able to produce a stable biofilm that adhered to polystyrene microwells. Thymus vulgaris essential oil produced inhibitory effects against all isolates with considerable variation in susceptibility. The minimum inhibitory concentration (MIC) and biofilm inhibitory concentration (BIC) values varied between 0.0625 to 2% v/v. Isolate 6 (Pseudomonas aeruginosa) was the most resistant for both planktonic and biofilm growth while isolate 4 Escherichia coli was the most sensitive. Isolate 6 was the only isolate to exhibit a specific biofilm-related response to Thymus vulgaris essential oil i.e. the minimum Thymus vulgaris essential oil concentration needed to inhibit biofilm adherence was much lower than that required to inhibit planktonic growth. For the remaining isolates, BIC for the biofilm did not differ from the MIC for planktonic. Thymus vulgaris essential oil was able to inhibit Pseudomonas aeruginosa adherence to polystyrene. Conclusions The MIC and BIC values of Thymus vulgaris essential oil show that the essential oil has strong antibacterial and antibiofilm activity. Thymus vulgaris was able to inhibit biofilm formation in the most tolerant isolate at sub-inhibitory concentrations.

There is a high variability in chemical composition of essential oil from Thymus vulgaris grown in different countries and different localities in the same country. Only three reports were published on the essential oil composition of Thymus vulgaris grown in Jordan and non of them reported from South Jordan. The current study aims to determine the essential oil composition of Thymus vulgaris grown wild in South Jordan and to test their activity against clinical isolate antibiotics resistant bacteria. The composition of the essential oil from Thymus vulgaris was determined using GC-MS while the screening for essential oil activity was carried out using disc diffusion method. The results of GC-MS produced forty-eight components of the oil representing more than 97% of the oil contents, monoterpenes being most abundant (about 85%) with thymol (37.05%), cis- dihydrocarvone (9.34%), carvacrol (8.45%), hydroxy-3-(3-methyl-2-butenyl)-3-cyclopenten-1-one (8.41%), p- cymene (5.73%), cis-sabinene (4.42%), z-isoeugenol (3.342%), and aromadendrene (3.42%) as major constituents. The essential oil from Thymus vulgaris was found to be active against all the tested bacteria except in Pseudomonas aeruginosa. The high concentrations of thymol and carvacrol may account for its high activity against resistant bacteria.

Data regarding the levels of inflammatory mediators in patients suffering of the rare disease Alkaptonuria are limited. C-reactive protein (CRP), Interleukin-1 Beta (IL-1β) and Interleukin-6 (IL-6) are acute-phase markers associated with joint inflammation. The aim of the present study is to compare the serum levels of the pro-inflammatory mediators, CRP, IL-1β and IL-6 in alkaptonuria patients (n = 17) with those measured for age-matched healthy controls (n = 17). Moreover, we attempt to determine the association between cytokine levels with the disease severity score and age using the Spearman correlation and multiple linear regression. The results show that the serum concentrations of the IL-1 β, IL-6 and CRP are higher in AKU patients compared with healthy controls, with a significant difference in IL-6 (p = 0.02). Moreover, a positive correlation is found between the patients' serum IL-6 and patients' age and the AKU Severity Score ( r o = 0.73 and 0.7, respectively; p < 0.05). Thus, the patients' IL-6 serum levels can predict the disease severity score in alkaptonuria patients (p < 0.05). These findings suggest that the IL-6 might play a role in the pathogenesis of inflammation in AKU patients and thus targeting it may be one mode of treatment in future. However, these findings need to be supported by further studies, conducted on a larger sample of patients.

Calponin is an actin- and calmodulin-binding protein believed to regulate the function of actin. Low-resolution studies based on proteolysis established that the recombinant calponin fragment 131-228 contained actin and calmodulin recognition sites but failed to precisely identify the actin-binding determinants. In this study, we used NMR spectroscopy to investigate the structure of this functionally important region of calponin and map its interaction with actin and calmodulin at amino-acid resolution. Our data indicates that the free calponin peptide is largely unstructured in solution, although four short amino-acid stretches corresponding to residues 140-146, 159-165, 189-195, and 199-205 display the propensity to form α-helices. The presence of four sequential transient helices probably provides the conformational malleability needed for the promiscuous nature of this region of calponin. We identified all amino acids involved in actin binding and demonstrated for the first time, to our knowledge, that the N-terminal flanking region of Lys(137)-Tyr(144) is an integral part of the actin-binding site. We have also delineated the second actin-binding site to amino acids Thr(180)-Asp(190). Ca(2+)-calmodulin binding extends beyond the previously identified minimal sequence of 153-163 and includes most amino acids within the stretch 143-165. In addition, we found that calmodulin induces chemical shift perturbations of amino acids 188-190 demonstrating for the first time, to our knowledge, an effect of Ca(2+)-calmodulin on this region. The spatial relationship of the actin and calmodulin contacts as well as the transient α-helical structures within the regulatory region of calponin provides a structural framework for understanding the Ca(2+)-dependent regulation of the actin-calponin interaction by calmodulin.