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Background: Colon cancer is the most aggressive form of cancers, that causes 0.5 million deaths per year around the globe. Targeting colon cancer by conventional therapeutic options elicits toxicity. Traditional medicines take a lead to alleviate the existing clinical challenges. Objective: To investigate antibacterial activity against Helicobacter Pylori and in vitro anti-colon cancer activity by Acacia nilotica extract (ACE) and its active constituent pyrogallol. Methods: Pyrogallol isolated from A. nilotica by column chromatography and HPLC and structure was elucidated by spectral analysis. Antibacterial activity was done by flow cytometry. Cytotoxicity was measured by MTT assay. Apoptotic morphology and nuclear fragmentation were assessed with AO/ethidium bromide and DAPI staining. DNA fragmentation was done by electrophoresis. Western blot used to analyze the molecular mechanism of apoptosis. Cell cycle arrest was determined using flow cytometry of propidium iodide stained cells. Cell migration was determined by wound healing assay. Results: ACE (20 µg/ml) and pyrogallol (10 µg/ml) treatment reduced the survival of H.pylori at 61% and 62%, respectively. MTT results show that HT-29 cells are more sensitive to pyrogallol with an IC50 value of 35μg/ml compared to ACE. Pyrogallol treated HT-29 cells reached dead state i.e. late apoptotic state with severe nuclear fragmentation. Pyrogallol elicits dose dependent DNA fragmentation in HT-29 cells. Pyrogallol induced apoptosis by simultaneous down-regulation of Bcl-2 and up-regulation of BAX and cytochrome c. Pyrogallol arrested HT-29 cells in S and G2/M phase of cell cycle. Further pyrogallol exhibited marked antimetastatic potential by inhibiting the migration of HT-29 cells dose dependently. Conclusion: Both ACE and pyrogallol repressed the growth of H.pylori and as significant anti-colon cancer agent.

Objective To explore whether a DNA immunization approach targeting the major haemorrhage molecule of a prothrombin activator-like metalloproteinase from Echis ocellatus ( E. ocellatus ) venom could be conceived to inspire antibodies with more prominent specificity and equal adequacy to current conventional antivenoms systems. Methods The isolated DNA EoMP-6 was used as the template for PCR amplification using the EoDC-2-specific forward and reverse primers. A PCR product of approximately 700 bp was obtained and cloned into pSecTag-B expression vector where anti-EoDC-2 antibodies were generated and analysed for their efficacy to neutralise local haemorrhage in vitro and in vivo . Results Our results suggest that the generated anti-EoDC-2 showed a remarkable efficacy by (a) interfering with the interaction of the recombinant disintegrin “EoDC-2” isolated from the E. ocellatus as well as other viper species to the α 2 β 1 -integrins on platelets; (b) complete inhibition of the catalytic site of the metalloproteinase molecules in vitro using an adaptation antibody zymography assay. Furthermore, it has a polyspecific potential and constitutively expressed significant inhibition by cross-reaction and neutralised venom-induced local haemorrhage exerted by different viper species in vivo . The potential characteristic of EoDC-2 against one part (the non-catalytic domain) as opposed to the whole molecule to neutralise its haemorrhagic activity is of crucial importance as it represents a novel approach with greater immunological specificity and fewer hazards, if any, than conventional systems of antivenom production, by exposure large animals that usually being used for the current antivenom production to a less injurious than expression of the whole molecule containing the catalytic metalloprotease domain. Hence, we report for the first time that our preliminary results hold a promising future for antivenom development. Conclusions Antibodies generated against the E. ocellatus venom prothrombin activator-like metalloprotease and disintegrin-cysteine-rich domains modulated and inhibited the catalytic activity both in vitro and in vivo of venom metalloproteinase disintegrin cysteine rich molecules. Thus, generating of venom specific-toxin antibodies by DNA immunization offer a more rational treatment of snake envenoming than conventional antivenom.

Breast cancer is a global health concern and is a major cause of death among women. In Oman, it is the most common cancer in women, with an incidence rate of 15.6 per 100,000 Omani females. Various anticancer remedies have been discovered from natural products in the past and the search is continuing for additional examples. Cytotoxic natural compounds may have a major role in cancer therapy either in potentiating the effect of chemotherapy or reducing its harmful effects. Recently, a few studies have reported advantages of using crude camel milk in treating some forms of cancer. However, no adequate data are available on the lyophilised camel's milk responsibility for triggering apoptosis and oxidative stress associated with human breast cancer. The present study aimed to address the role of the lyophilised camel's milk in inducing proliferation repression of BT-474 and HEp-2 cells compared with the non-cancer HCC1937 BL cell line. Lyophilized camel's milk fundamentally repressed BT-474 cells growth and proliferation through the initiation of either the intrinsic and extrinsic apoptotic pathways as indicated by both caspase-3 mRNA and its action level, and induction of death receptors in BT-474 but not the HEp-2 cell line. In addition, lyophilised camel's milk enhanced the expression of oxidative stress markers, heme-oxygenase-1 and reactive oxygen species production in BT-474 cells. Increase in caspase-3 mRNA levels by the lyophilised camel's milk was completely prevented by the actinomycin D, a transcriptional inhibitor. This suggests that lyophilized camel's milk increased newly synthesized RNA. Interestingly,it significantly (p

Objective To investigate the levels of zinc-α-2-glycoprotein (ZAG) among Omani AIDS patients receiving combined antiretroviral therapy (cART). Methods A total of 80 Omani AIDS patients (45 males and 35 females), average age of 36 years, who were receiving cART at the Sultan Qaboos University Hospital (SQUH), Muscat, Oman, were tested for the levels of ZAG. In addition, 80 healthy blood donors (46 males and 34 females), average age of 26 years, attending the SQUH Blood Bank, were tested in parallel as a control group. Measurement of the ZAG levels was performed using a competitive enzyme-linked immunosorbent assay and in accordance with the manufacturer's instructions. Results The ZAG levels were found to be significantly higher among AIDS patients compared to the healthy individuals (P=0.033). A total of 56 (70%) of the AIDS patients were found to have higher levels of ZAG and 16 (20%) AIDS patients were found to have high ZAG levels, which are significantly (P>0.031) associated with weight loss. Conclusions ZAG levels are high among Omani AIDS patients on cART and this necessitates the measurement of ZAG on routine basis, as it is associated with weight loss.

C-C motif chemokine receptor-5 (CCR5) is a pro-inflammatory receptor that binds to chemokines and facilitates the entry of the R5 strain of HIV-1. A number of polymorphisms were identified within the promoter and coding regions of the CCR5 gene, some of which have been found to affect the protein expression and thus receptor function. Although several CCR5 polymorphisms were shown to vary widely in their distribution among different ethnic populations, there has been no study addressing the potential variants of the CCR5 gene in the Omani population. The aim of this study was to identify the polymorphic sites that exist within the CCR5 gene in Omanis. Blood samples were collected from 89 Omani adult individuals, and genomic DNA was amplified by polymerase chain reaction and sequenced to identify the polymorphic sites. The distribution of the detected variants was examined and compared with the previously published data. Four new indels were detected of 32 variable positions, -2973A/-, -2894A/-, -2827TA/- and -2769T/-, and all were located in the 5'UTR. Furthermore, two new mutations, -2248G/A and +658A/G, were observed for the first time; the -2248G/A was detected in the intron 1 region in one subject and +658A/G in the coding region of the CCR5 in another subject. In silico analysis showed that the novel variations in the 5'UTR may have effects on the transcription factor binding sites. Therefore, this study demonstrates the presence of two new SNPs and four novel indels in the CCR5 gene in the Omani population. Our findings support the wide spectrum of genetic diversity reported within the CCR5 gene region among different ethnic groups.

Laboratory testing is of great value when evaluating a patient with a suspected autoimmune disease. The results can confirm a diagnosis, estimate disease severity, aid in assessing prognosis and are useful to follow disease activity. Components of the laboratory exam include complete blood count with differential, comprehensive metabolic panel, inflammatory markers, autoantibodies, and flow cytometry. Currently, autoimmunity laboratories are very vibrant owing to the constant and increasing availability of new tests, mainly due to the detection of new autoantibodies. The main characteristic that differentiates the autoimmunity laboratory from other laboratories is the use of immunoassays such as enzyme-linked immunosorbent assay (ELISA), as basic techniques which determines antibodies (autoantibodies) and not antigens. For this reason, immunoassay techniques must employ antigens as reagents. However, over the last few years, a significant trend at autoimmunity laboratories has been the gradual replacement of immunofluorescence microscopy by immunoassay. Nowadays the revolution of new technology has taken place significantly, for examples; recombinant DNA technology has allowed the production of large quantities of antigens for autoantibody analysis. Flow cytometry for the analysis of microsphere-based immunoassays allows the simultaneous measurement of several autoantibodies. In the same way, autoantigen microarrays provide a practical means to analyse biological fluids in the search for a high number of autoantibodies. We are now at the beginning of an era of multiplexed analysis, with a high capacity of autoantibody specificities. The future tendency in this field will include immunoassays with greater analytical sensitivity, specificity, simultaneous multiplexed capability, the use of protein microarrays, and the use of other technologies such as microfluidics.

Objective To study in vitro antibacterial and antifungal activity of hot water and methanolic extracts of the three medicinal plants- Boswellia (Luban) species. Methods Three selected plants were collected from different localities of Soqotra (Republic of Yemen), Dohfar (Sultanate of Oman) and Republic of Somalia. The plants were dried and extracted with two different solvents (methanol and hot water) to yield six crude extracts. The obtained extracts were tested for their antibacterial activity against eleven different bacterial strains and two fungi using the standard well-diffusion and micro-dilution methods. The following microorganisms were used: methicillin-resistant Staphylococcus aureus (ATCC 6538), muti-drug resistant Pseudomonas aeruginosa (ATCC 27853), enterohemorrhagic Escherichia coli (0157 EHEC), Salmonella typhi, Proteus vulgaris, Klebsiella pneumoniae, Bacillus subtilus (ATCC 6059, reference strain), Streptococcus pneumoniae, Klebsella pneumonia, MRSA, Corynebacterium, Corynebacterium diphtheriae and two fungus: Candida maltosa and Candida albicans . Results The different extracts possessed different inhibitory activity against different types of bacterial species. The patterns of inhibition varied with the plant extract, the solvent used for extraction and the organisms tested. The antimicrobial activity exhibited by the methanolic extracts of Boswellia sacra from the Suqotra and Dhofar regions was greater than that of Boswellia frereana collected from Somalia. The methanolic extract of the oleo-gum-resin showed higher efficacy to inhibit all the tested bacterial strains than the methanolic extract of frankincense-resin . The Boswellia frereana collected from Somalia showed lower activity compared with the two other Boswellia species. The plant extracts showed bacteriostatic activity at lower concentrations and bactericidal activity at higher concentrations. Neither water nor methanolic extracts showed any activity against the fungi Candida maltosa and Candida albicans . Conclusions It can be concluded that the methanolic extracts of Boswellia (Luban) possess the highest antibacterial activity. Neither water nor methanolic extracts show any activity against Candida maltosa and Candida albicans.

Traditional medicine is still widely practiced in many countries due to its complexity and long term benefit. Among various medicinal plants found in Dhofar region of Oman, frankincense has a unique position due to its medicinal and economical importance. The gum-resin and essential oil produced by different species, most of these are related to family Burseraceae under the Boswellia genus. The family members of Boswellia are characterized by resin bearing ducts. Among the twenty-nine species of genus Boswellia, (Burseraceae), Boswellia sacra Flueck is known since decades for the extraction of aromatic gums and resins that are burned as incense. Boswellia resin holds about 60-80% alcohol-soluble resin, 15-20% water soluble gum and 5-7 % essential oil, along with polysaccharide fraction and polymeric substances are also present in limited extent. The physicians and nutritionists show interest in frankincense due to the therapeutic potential of its gum resin and essential oil. Essential oil and gum resin of various species of frankincense has been used to make remedies to treat different diseases. Various reports have described the antimicrobial, immuno-modulatory, anti-inflammatory, hypoglycemic, anticancer, anti-asthmatic, antidiarrheal, hypo-lipidemic, anti-diabetic, hepato-protective, and even antiviral effect of different Boswellia species. In this review, we have highlighted the works done so far on the use of Boswellia gum resin and health benefits along with some pilot clinical studies done on the Boswellia gum resin related products.

The traditional medicine still plays an important role in the primary health care in Yemen. The current study represents the investigation of 16 selected plants, which were collected from different localities of Yemen. The plants were dried and extracted with two different solvents (methanol and hot water) to yield 34 crude extracts. The obtained extracts were tested for their antimicrobial activity against three Gram-positive bacteria, two Gram-negative bacteria, one yeast species and three multiresistantStaphylococcusstrains using agar diffusion method, for their antioxidant activity using scavenging activity of DPPH radical method and for their cytotoxic activity using the neutral red uptake assay. In addition, a phytochemical screening of the methanolic extracts was done. Antibacterial activity was shown only against Gram-positive bacteria, among them multiresistant bacteria. The highest antimicrobial activity was exhibited by the methanolic extracts ofAcalypha fruticosa,Centaurea pseudosinaica,Dodonaea viscosa,Jatropha variegata,Lippia citriodora,Plectranthus hadiensis,Tragia pungensandVerbascum bottae.Six methanolic extracts especially those ofA. fruticosa,Actiniopteris semiflabellata,D. viscosa,P. hadiensis,T. pungensandV. bottaeshowed high free radical scavenging activity. Moreover, remarkable cytotoxic activity against FL-cells was found for the methanolic extracts ofA. fruticosa,Iris albicans,L. citriodoraandT. pungens. The phytochemical screening demonstrated the presence of different types of compounds like flavonoids, terpenoids and others, which could be responsible for the obtained activities.

The objective of the study is to investigate whether theHibiscus aethiopicusL. plant has neutralization activity against venoms of two clinically important snakes. TheH. aethiopicuswas dried and extracted with water. Different assays were performed to evaluate the plant's acute toxicity and its anti-snake venom activities. The results showed thatH. aethiopicusextract alone had no effect on the viability of C2C12muscle cells, but significantly (P<.05) protected muscle cells against the toxic effects ofE. ocellatusvenom at 55, 150, and 300 μg/mL. The maximum protective effect of the extract was exhibited at 75 μg/mL. The extract significantly (P<.001) inhibited the cytotoxic effects ofE. ocellatusvenom at 300 μg/mL. All rabbits (n=10) and guinea pigs (n=10) were alive after the two weeks of given the lethal dosage 16 g/Kg of theH. aethiopicusextract herbal solution. No abnormal behaviour was observed of both groups of animals. All guinea pigs (n=3) treated with venoms alone (5 mg/kg) died. However, all guinea pigs (n=21) treated with venom (5 mg/kg) and the extract (400 to 1000 mg/kg) survived. Guinea pigs (n=3) treated withNaja n. nigricollisvenom alone (2.5 mg/kg) and guinea pigs (n=21) venom with the extract (400 to 1000 mg/kg) died. TheH. aethiopicuscompletely (100%) blocked the haemorrhagic activity ofE. ocellatusin the egg embryo at 3.3 mg/mL of extract. These findings suggest thatH. aethiopicusmay contain an endogenous inhibitor of venom-induced haemorrhage.

Objective To estimate the rate of occult hepatitis B virus (HBV) among patients with chronic liver disease (CLD). Methods After an informed consent, sera samples were collected during April 2004 to April 2005 from 280 patients (200 male and 80 female). They were previously diagnosed with CLD based on history and ultrasound and were investigated for occult HBV infection. Sera were first screened for HBsAg and those which showed negative were tested for anti-HBc. The anti-HBc positive sera were further tested for anti-HBs to identify sera with isolated anti-HBc which in turn were subjected to HBV–DNA testing using PCR to determine the rate of occult HBV infection. Moreover, sera with occult HBV were tested for Anti-HCV and HCV-RNA using RT-PCR. Results HBsAg was detected in 44 of 280 (15.7%). Of 236 HBsAg negative sera anti-HBc was detected in 22 (9.3%). All anti-HBc positive sera were found to be anti-HBs negative. HBV–DNA was detected in 11 of 22 (50.0%) sera with isolated anti-HBc indicating occult HBV in 4.3% of all sera. None of the sera with occult HBV had anti-HCV or HCV-RNA. Conclusions Occult HBV infection does exist among CLD patients in Yemen and the mechanism of its occurrence merits further investigation.

The antimicrobial activity of the ethanol extract of the Auklandia (Saussurea lappa)root plant was investigated to verify its medicinal use in the treatment of microbial infections.The antimicrobial activity of the ethanol extract was tested against clinical isolates of some multidrug-resistant bacteria using the agar well diffusion method. Commercial antibiotics were used as positive reference standards to determine the sensitivity of the clinical isolates.The extracts showed significant inhibitory activity against clinical isolates of methicillin resistant Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumonia, Extended Spectrum Beta-Lactemase, Acinetobacter baumannii. The minimum inhibitory concentration values obtained using the agar dilution test ranged from 2.0 µg/µL-12.0 µg/µL. In the contrary the water extract showed no activity at all against the tested isolates. Furthermore, the results obtained by examining anti-resistant activity of the plant ethanolic extract showed that at higher concentration of the plant extract (12 µg) all tested bacteria isolates were inhibited with variable inhibition zones similar to those obtained when we applied lower extract concentration using the well diffusion assay.The results demonstrated that the crude ethanolic extract of the Auklandia (Saussurea lappa) root plant has a wide spectrum of activity suggesting that it may be useful in the treatment of infections caused by the above clinical isolates (human pathogens).

The objective of the study is to investigate the anti-snake venom activities of a local plant,Hibiscus aethiopicusL. TheH. aethiopicuswas dried and extracted with ethanol. Different assays were performed according to standard techniques, to evaluate the plant’s acute toxicity and its antivenom activities. The results of evaluating the systemic acute toxicity of theH. aethiopicusextract using “oral and intra-peritoneal” route were normal even at the highest dose (24 g/kg) tested. All guinea pigs (n=3) when treated with venomsE. c. sochureki(75 μg) alone induced acute skin haemorrhage. In contrast, all guinea pigs (n=18) treated with both venom and the plant extract at a concentration between 500 and 1000 mg/kg showed no signs of haemorrhage. Moreover, all guinea pigs (n=18) treated with venom and the plant extract below 400 mg/kg showed acute skin haemorrhage. All guinea pigs treated with venomE. c. sochureki(75 μg) alone induced acute skin haemorrhage after both 24 and 32 hours. In contrast, all guinea pigs treated with both venom and the plant extract (administered independently) at concentrations between 500 and 1000 mg/kg showed no signs of haemorrhage after 32 hours. However, after 24 hours all tested guinea pigs showed less inhibition (H. aethiopicusplant may contain an endogenous inhibitor of venom induced local haemorrhage.

To investigate the antibacterial activity of henna (Lawsonia inermis Linn) obtained from different regions of Oman against a wide array of micro-organisms.Fresh henna samples were obtained from different regions of Oman as leaves and seeds. 100 g fresh and dry leaves and 50 g of fresh and dry seeds were separately soaked in 500 mL of ethanol for three days, respectively, with frequent agitation. The mixture was filtered, and the crude extract was collected. The crude extract was then heated, at 48 °C in a water bath to evaporate its liquid content. The dry crude henna extract was then tested for its antibacterial activity using well-diffusion antibiotic susceptibility technique. Henna extracts were investigated for their antibacterial activity at different concentrations against a wide array of different micro-organisms including a laboratory standard bacterial strain of Pseudomonas aeruginosa (NCTC 10662) (P. aeruginosa) and eleven fresh clinical isolates of P. aeruginosa obtained from patients attending the Sultan Qaboos University Hospital (SQUH). 2-Hydroxy-p-Nathoqinone-Tech (2-HPNT, MW=174.16, C10H6O3) was included as control (at 50% concentration) along with the henna samples tested.Henna samples demonstrated antibacterial activity against all isolates but the highest susceptibility was against P. aeruginosa with henna samples obtained from Al-sharqyia region.Omani henna from Al-sharqyia region demonstrates high in vitro anti-P. aeruginosa activity compared with many henna samples from different regions of Oman.

Envenoming by Echis saw scaled vipers and Bitis arietans puff adders is the leading cause of death and morbidity in Africa due to snake bite. Despite their medical importance, the composition and constituent functionality of venoms from these vipers remains poorly understood. Here, we report the cloning of cDNA sequences encoding seven clusters or isoforms of the haemostasis-disruptive C-type lectin (CTL) proteins from the venom glands of Echis ocellatus, E. pyramidum leakeyi, E. carinatus sochureki and B. arietans. All these CTL sequences encoded the cysteine scaffold that defines the carbohydrate-recognition domain of mammalian CTLs. All but one of the Echis and Bitis CTL sequences showed greater sequence similarity to the beta than alpha CTL subunits in venoms of related Asian and American vipers. Four of the new CTL clusters showed marked inter-cluster sequence conservation across all four viper species which were significantly different from that of previously published viper CTLs. The other three Echis and Bitis CTL clusters showed varying degrees of sequence similarity to published viper venom CTLs. Because viper venom CTLs exhibit a high degree of sequence similarity and yet exert profoundly different effects on the mammalian haemostatic system, no attempt was made to assign functionality to the new Echis and Bitis CTLs on the basis of sequence alone. The extraordinary level of inter-specific and inter-generic sequence conservation exhibited by the Echis and Bitis CTLs leads us to speculate that antibodies to representative molecules should neutralise the biological function of this important group of venom toxins in vipers that are distributed throughout Africa, the Middle East and the Indian subcontinent. (C) 2003 Elsevier B.V. All rights reserved.

This review focuses on DNA vaccines, denoting the last two decades since the early substantiation of preclinical protection was published in Science in 1993 by Ulmer et al. In spite of being safely administered and easily engineered and manufactured DNA vaccine, it holds the future prospects of immunization by inducing potent cellular immune responses against infectious and non-infectious diseases. It is well documented that injection of DNA plasmid encoding a desired gene of interest can result in the subsequent expression of its products and lead to the induction of an immune response within a host. This is pertinent to prophylactic and therapeutic vaccination approach when the peculiar gene produces a protective epitope from a pathogen. The recent studies demonstrated by a number of research centers showed that these immune responses evoke protective immunity against several infectious diseases and cancers, which provides adequate support for the use of this approach. We attempt in this review to provide an informative and unbiased overview of the general principles and concept of DNA vaccines technology with a summary of a novel approach to the DNA vaccine, present investigations that describe the mechanism(s) of protective immunity provoked by DNA immunization and to highlight the advantages and disadvantages of DNA immunisation.

Objective: To investigate different types of dates and medical properties of influencing blood clotting and wound healing in an animal model. Methods: Three different cultivars of dates (Ajwa, Khalas, and Fardh) were examined in-vivo, for blood clotting and wound healing using CD1 mice of both sexes. Study of toxicity to animals was performed accordingly prior to further investigations. The ethanolic extracts were given orally to animals as a constituent in their daily water. Blood samples were obtained from the mice inferior vena cava to carry out the prothrombin time (PT) assay using the manual method and confirmed using a semi-automated machine. The bleeding time (BT) assay was performed using the cutting technique. In the wound healing analysis, a small cut (5-10 mm) in the skin overlying the thigh was conducted in all mice under anesthesia. The diameter of the cut and healing status were measured on a daily basis throughout the time of the experiment using a roller. Results: Ajwa was able to elevate both PT and BT (P