Sami A. Alyahya

Proteomic Analysis of the Protein Expression Profile in the Mature Nigella sativa (Black Seed)

Nigella sativa (N. sativa) seed has been used as an important nutritional flavoring agent and in traditional medicine for treating many illnesses since ancient times. Understanding the proteomic component of the seed may lead to enhance the understanding of its structural and biological functional complexity. In this study, we have analyzed its proteome profile based on gel-based proteome mapping technique that includes one-dimensional gel electrophoresis followed by liquid chromatography and tandem mass spectrometry strategy. We have not come across any such studies that have been performed in N. sativa seeds up to date. A total of 277 proteins were identified, and their functional, metabolic, and location-wise annotations were carried out using the UniProt database. The majority of proteins identified in the proteome dataset based on their function were those involved in enzyme catalytic activity, nucleotide binding, and protein binding while the major cellular processes included regulation of biological process followed by regulation of secondary biological process, cell organization and biogenesis, protein metabolism, and transport. The identified proteome was localized mainly to the nucleus then to the cytoplasm, plasma membrane, mitochondria, plastid, and others. A majority of the proteins were involved in biochemical pathways involving carbohydrate metabolism, amino acid and shikimate pathway, lipid metabolism, nucleotide, cell organization and biogenesis, transport, and defense processes. The identified proteins in the dataset help to improve our understanding of the pathways involved in N. sativa seed metabolism and its biochemical features and detail out useful information that may help to utilize these proteins. This study could thus pave a way for future further high-throughput studies using a more targeted proteomic approach.

Histopathological studies of experimental Aeromonas hydrophila infection in blue tilapia, Oreochromis aureus

Blue tilapia, Oreochromis aureus, was experimentally infected with Aeromonas hydrophila, a bacterium that damages the gills, liver, and intestine, resulting in histopathological changes in the infected organs. Our histopathological study showed an aggregation of hemocytes with cell necrosis in gills; a massive aggregation of hemocytes and pyknotic nuclei in the hepatopancreas; and a lower rate of hemocyte aggregation in the digestive system of the infected fish.

Development of self-repair nano-rod scaffold materials for implantation of osteosarcoma affected bone tissue

Osteosarcoma is the most widely recognized fatal bone disease in children and young adults. The osteosarcoma affected places of bone implant materials lose their activity after a period of time due to the possibility of regenerating sarcoma cells. Hence, the complete recovery of this disease is very challenging. Subsequently, new helpful methodologies, including natural antioxidant loaded bone implant materials, are effectively used to treat osteosarcoma cells. In this regard, nano-hydroxyapatite reinforced with a xylitol based poly(xylitol sebacate) PXS co-polymer together with a capsaicin loaded scaffold was investigated on osteosarcoma cells. The physicochemical properties of the scaffold were evaluated by FT-IR (Fourier transform infrared spectroscopy), SEM (scanning electron microscopy), TEM (transmission electron microscopy), and XRD (X-ray diffraction). The in vitro release and antioxidant activity of the capsaicin loaded nHAP/PXS/CAP scaffold were evaluated by UV-Visible spectroscopy. Cytotoxicity against the Saos-2 cancer line and cell viability in the osteoblast cell MG63 are reported. Eventually, the composite enlarges the creation of reactive oxygen species (ROS) in Saos-2 cells.

Proteomic Profiling Comparing the Effects of Different Heat Treatments on Camel (Camelus dromedarius) Milk Whey Proteins

Camel milk is consumed in the Middle East because of its high nutritional value. Traditional heating methods and the duration of heating affect the protein content and nutritional quality of the milk. We examined the denaturation of whey proteins in camel milk by assessing the effects of temperature on the whey protein profile at room temperature (RT), moderate heating at 63 °C, and at 98 °C, for 1 h. The qualitative and quantitative variations in the whey proteins before and after heat treatments were determined using quantitative 2D-difference in gel electrophoresis (DIGE)-mass spectrometry. Qualitative gel image analysis revealed a similar spot distribution between samples at RT and those heated at 63 °C, while the spot distribution between RT and samples heated at 98 °C differed. One hundred sixteen protein spots were determined to be significantly different (p < 0.05 and a fold change of ≥1.2) between the non-heated and heated milk samples. Eighty protein spots were decreased in common in both the heat-treated samples and an additional 25 spots were further decreased in the 98 °C sample. The proteins with decreased abundance included serum albumin, lactadherin, fibrinogen β and γ chain, lactotransferrin, active receptor type-2A, arginase-1, glutathione peroxidase-1 and, thiopurine S, etc. Eight protein spots were increased in common to both the samples when compared to RT and included α-lactalbumin, a glycosylation-dependent cell adhesion molecule. Whey proteins present in camel milk were less affected by heating at 63 °C than at 98 °C. This experimental study showed that denaturation increased significantly as the temperature increased from 63 to 98 °C.

Swift fabrication of Ag nanostructures using a colloidal solution of Holostemma ada-kodien (Apocynaceae) – Antibiofilm potential, insecticidal activity against mosquitoes and non-target impact on water bugs

Recent research in entomology and parasitology focused on the efficacy of green fabricated nanomaterials as novel insecticides. In this study, we synthesized poly-dispersed and stable silver nanoparticles (AgNPs) using the leaf extract of Holostemma ada-kodien. The nanostructures were characterized by ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, energy dispersive X-ray, and X-ray diffraction analysis. The efficacy of H. ada-kodien leaf extract and AgNPs in vector control was evaluated against the mosquitoes Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus, which act as major vectors of important parasitic and arboviral diseases. AgNPs showed higher toxicity if compared to the H. ada-kodien leaf aqueous extract, LC50 towards larvae of A. stephensi, A. aegypti, and C. quinquefasciatus were 12.18, 13.30, and 14.70 μg/mL, respectively. When the AgNPs were tested on non-target water bugs, Diplonychus indicus, the LC50 value was 623.48 μg/mL. Furthermore, 100 μl/mL of AgNPs achieved significant antimicrobial activity against Bacillus pumilus, Enterococcus faecalis, Pseudomonas aeruginosa, Proteus vulgaris, and Candida albicans. Light and confocal laser scanning microscopy highlighted a major impact of the H. ada-kodien-synthesized AgNPs on the external topography and architecture of microbial biofilms, both on Gram-positive and Gram-negative bacteria. Overall, this study sheds light on the insecticidal and antibiofilm potential of H. ada-kodien-synthesized AgNPs, a potential green resource for the rapid synthesis of polydispersed and highly stable AgNPs.

Computational systems biology analysis of biomarkers in lung cancer; unravelling genomic regions which frequently encode biomarkers, enriched pathways, and new candidates

Exponentially growing scientific knowledge in scientific publications has resulted in the emergence of a new interdisciplinary science of literature mining. In text mining, the machine reads the published literature and transfers the discovered knowledge to mathematical-like formulas. In an integrative approach in this study, we used text mining in combination with network discovery, pathway analysis, and enrichment analysis of genomic regions for better understanding of biomarkers in lung cancer. Particular attention was paid to non-coding biomarkers. In total, 60 MicroRNA biomarkers were reported for lung cancer, including some prognostic biomarkers. MIR21, MIR155, MALAT1, and MIR31 were the top non-coding RNA biomarkers of lung cancer. Text mining identified 447 proteins which have been studied as biomarkers in lung cancer. EGFR (receptor), TP53 (transcription factor), KRAS, CDKN2A, ENO2, KRT19, RASSF1, GRP (ligand), SHOX2 (transcription factor), and ERBB2 (receptor) were the most studied proteins. Within small molecules, thymosin-a1, oestrogen, and 8-OHdG have received more attention. We found some chromosomal bands, such as 7q32.2, 18q12.1, 6p12, 11p15.5, and 3p21.3 that are highly involved in deriving lung cancer biomarkers.

Insecticidal activity of camphene, zerumbone and α-humulene from Cheilocostus speciosus rhizome essential oil against the Old-World bollworm, Helicoverpa armigera

The fast-growing resistance development to several synthetic and microbial insecticides currently marketed highlighted the pressing need to develop novel and eco-friendly pesticides. Among the latter, botanical ones are attracting high research interest due to their multiple mechanisms of action and reduced toxicity on non-target vertebrates. Helicoverpa armigera (Lepidoptera: Noctuidae) is a key polyphagous insect pest showing insecticide resistance to several synthetic molecules used for its control. Therefore, here we focused on the rhizome essential oil extracted from an overlooked Asian plant species, Cheilocostus speciosus (J. Konig) C. Specht (Costaceae), as a source of compounds showing ingestion toxicity against H. armigera third instar larvae, as well as ovicidal toxicity. In acute larvicidal assays conducted after 24h, the C. speciosus essential oil achieved a LC50 value of 207.45µg/ml. GC and GC-MS analyses highlighted the presence of zerumbone (38.6%), α-humulene (14.5%) and camphene (9.3%) as the major compounds of the oil. Ingestion toxicity tests carried out testing these pure molecules showed LC50 values of 10.64, 17.16 and 20.86µg/ml, for camphene, zerumbone and α-humulene, respectively. Moreover, EC50 values calculated on H. armigera eggs were 35.39, 59.51 and 77.10µg/ml for camphene, zerumbone and α-humulene, respectively. Overall, this study represents the first report on the toxicity of C. speciosus essential oil against insect pests of agricultural and medical veterinary importance, highlighting that camphene, zerumbone and α-humulene have a promising potential as eco-friendly botanical insecticides.

Antibiotic-resistant Staphylococcus epidermidis isolated from patients and healthy students comparing with antibiotic-resistant bacteria isolated from pasteurized milk

Antibiotic-resistant Staphylococci are a global issue affecting humans, animals, and numerous natural environments. Antibiotic-resistant Staphylococcus epidermidis is an opportunistic pathogen frequently isolated from patients and healthy individuals. This study aimed to examine the antibiotic resistance of S. epidermidis isolated from patients, healthy students and compare the results with antibiotic-resistant bacteria isolated from pasteurized milk. Clinical strain isolation was performed in several hospitals in the Riyadh. Skin swabs from 100 healthy undergraduate candidate students were obtained at King Saud University. The pasteurized milk samples were obtained from local market (company, X). After isolation, identification and susceptibility tests were performed using an automated system. A multiplex tuf gene-based PCR assay was used to confirm identification. Biofilm production and biofilm-related gene expression were studied. S. epidermidis represented 17% of clinical bacterial isolates, and 1.7% of isolates obtained from healthy students were multiantibiotic-resistant. All patient strains were teicoplanin- and vancomycin-susceptible, while all student strains were gentamicin-, levofloxacin-, moxifloxacin-, and trimethoprim/sulfamethoxazole-susceptible. All the bacteria isolated from pasteurized milk were benzylpenicillin and oxacillin-resistant strains. Of the S. epidermidis strains, 91% could produce biofilms, and mecA, icaADBR, ica-ADB, ica-AD, ica-A only, and ica-C only were expressed in 83, 17.1, 25.7, 37.1, 20, and 0% of the strains, respectively. This work demonstrates that S. epidermidis can be accurately identified using a multiplex tuf-based assay, and that multiantibiotic-resistant S. epidermidis strains are widespread amongst patients and healthy students.

Nanosilver crystals capped with Bauhinia acuminata phytochemicals as new antimicrobials and mosquito larvicides

To develop novel nanoformulated insecticides and antimicrobials, herein we produced Ag nanoparticles (AgNPs) using the Bauhinia acuminata leaf extract. This unexpensive aqueous extract acted as a capping and reducing agent for the formation of AgNPs. We characterized B. acuminata-synthesized AgNPs by UV-vis and FTIR spectroscopy, XRD and TEM analyses. UV-vis spectroscopy analysis of B. acuminata-synthesized AgNPs showed a peak at 441.5 nm. FTIR shed light on functional groups from the phytoconstituents involved in nanosynthesis. XRD of B. acuminata-synthesized AgNPs suggested a face-centered cubic structure, with a highly crystalline nature. TEM of B. acuminata-synthesized AgNPs revealed mean size of 25 nm, with round shape. AgNPs tested at 60 μg/mL inhibited the growth of 5 bacteria and 3 fungal pathogens. In the insecticidal assays on important mosquito species, LC50 of the aqueous extract of B. acuminata leaves on the larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus were 204.07, 226.02, and 249.24 μg/mL, respectively. The B. acuminata-synthesized AgNPs exhibited higher larvicidal efficacy, with LC50 values of 24.59, 27.19, and 30.19 μg/mL, respectively. Therefore, herein we developed a single-step, reliable, inexpensive, and environmentally non-toxic synthesis process to obtain AgNPs with high bioactivity against pathogens and vectors. Given the effective antimicrobial and larvicidal activity, nanoparticles fabricated using plant extracts and extremely low concentrations of trace elements, such as silver, can be exploited for multipurpose activities. Our results pointed out that B. acuminata-synthesized AgNPs have a promising potential in antimicrobial food packaging, as well as a foliar spray to control plant pathogens in the field, and to synergize the efficacy of fungicidal and larvicidal formulations.

Biological Evaluation of Different Extracts of Aerial Parts of Nepeta deflersiana and Standardization of Active Extracts Using 8-Epi-7-Deoxyloganic Acid and Ursolic Acid by Validated HPTLC Method

Nepeta deflersiana (Lamiaceae) is a well-known medicinal plant that grows in Saudi Arabia. This plant is used in Saudi and Yemeni folk medicine as an anti-inflammatory, carminative, and antirheumatic agent. In order to prove its use in folk medicine, four different extracts from the aerial parts of the plant: petroleum ether, chloroform, ethyl acetate, and n-butanol extracts were subjected to biological assays to screen PPARα and PPARϒ agnostic, antioxidant, anti-inflammatory, and cytotoxic activities. Ethyl acetate and n-butanol extracts of N. deflersiana NDEE and NDBE, respectively, showed a decrease in oxidative stress and inhibition of both NF-kB and iNOS activities with no cytotoxic effects on four human cancer cell lines. Both active extracts were standardized using two bioactive metabolites which were isolated from the aerial parts of the same plant [8-epi-7-deoxyloganic acid (compound 1) and Ursolic acid (compound 2)] by developing a validated HPTLC method. It was found to provide a sharp and compact band of compound 1 at Rf = 0.07 and Rf = 0.57 for compound 2, using chloroform, methanol, and formic acid (8.9:0.8:0.3, v/v/v) as mobile phase at 550 nm. Compounds 1 and 2 were found in NDEE by 9.59 %, w/w, and 84.63 %, w/w, respectively, and by 11.97 %, w/w, and 21.26 %, w/w, respectively, in NDBE.