The Marine-Derived Sipholenol A-4-O-3′,4′-Dichlorobenzoate Inhibits Breast Cancer Growth and Motility in Vitro and in Vivo through the Suppression of Brk and FAK Signaling

Sipholenol A is a natural sipholane triterpenoid isolated from the Red Sea sponge, Callyspongia siphonella. Previous studies showed the antimigratory and antiproliferative activities of the semisynthetic sipholenol A esters against breast cancer cell lines. This study investigated the effects of sipholenol A-4-O-3′,4′-dichlorobenzoate (SPA) on the growth, migration and invasion of diverse human breast cancer cells. Results showed that SPA inhibited the growth of the human breast cancer cells, MDA-MB-231, MCF-7, BT-474 and T-47D, in a dose-dependent manner. Immunofluorescent analysis showed that SPA significantly reduced Ki-67-positive cells in MDA-MB-231 cells. Flow cytometry and Western blot analyses revealed that SPA treatment suppressed MDA-MB-231 cell growth by inducing cell cycle arrest at the G1 phase. In addition, SPA suppressed breast cancer cell migration, invasion and decreased Brk and FAK activation in a dose-dependent manner. Molecular docking study suggested a perfect fitting at the FAK’s FERM domain, inhibiting the main autophosphorylation site, Y397, which was further confirmed by Western blot analysis. Most known small molecule FAK inhibitors target the kinase domain, creating several off-target side effects. The in vivo studies showed that SPA treatment suppressed breast tumor growth and Ki-67, CD31, p-Brk and p-FAK expression in orthotopic breast cancer in nude mice. In conclusion, SPA inhibited the growth, invasion and migration of breast cancer cells possibly via deactivating Brk and FAK signaling, suggesting good potential for therapeutic use to control invasive breast cancer.     

Physiological and Molecular Analysis of Applied Nitrogen in Rice Genotypes

Ten genotypes of rice (Oryza sativa L.) were grown for 30 d in complete nutrient solution with 1 mmol/L (N-insufficient),4 mmol/L (N-moderate) and 10 mmol/L (N-high) nitrogen levels,and nitrogen efficiency (NE) was analyzed.Growth performance,measured in terms of fresh weight,dry weight and lengths of root and shoot,was higher in N-efficient than in N-inefficient rice genotypes at low N level.Of these 10 genotypes,Suraksha was identified as the most N-efficient,while Vivek Dhan the most N-inefficient.To find out the physiological basis of this difference,the nitrate uptake rate of root and the activities of nitrate assimilatory enzymes in leaves of N-efficient and N-inefficient rice genotypes were studied.Uptake experiments revealed the presence of two separate nitrate transporter systems mediating high-and low-affinity nitrate uptake.Interestingly,the nitrate uptake by the roots of Suraksha is mediated by both high-and low-affinity nitrate transporter systems,while that of Vivek Dhan by only low-affinity nitrate transporter system.Study of the activities and expression levels of nitrate assimilatory enzymes in N-efficient and N-inefficient rice genotypes showed that nitrate reductase (NR) and glutamine synthetase (GS) play important roles in N assimilation under low-nitrogen conditions.     

Biological Activities and Chemical Composition of Methanolic Extracts of Selected Autochthonous Microalgae Strains from the Red Sea

Four lipid-rich microalgal species from the Red Sea belonging to three different genera (Nannochloris, Picochlorum and Desmochloris), previously isolated as novel biodiesel feedstocks, were bioprospected for high-value, bioactive molecules. Methanol extracts were thus prepared from freeze-dried biomass and screened for different biological activities. Nannochloris sp. SBL1 and Desmochloris sp. SBL3 had the highest radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl, and the best copper and iron chelating activities. All species had potent butyrylcholinesterase inhibitory activity (>50%) and mildly inhibited tyrosinase. Picochlorum sp. SBL2 and Nannochloris sp. SBL4 extracts significantly reduced the viability of tumoral (HepG2 and HeLa) cells with lower toxicity against the non-tumoral murine stromal (S17) cells. Nannochloris sp. SBL1 significantly reduced the viability of Leishmania infantum down to 62% (250 µg/mL). Picochlorum sp. SBL2 had the highest total phenolic content, the major phenolic compounds identified being salicylic, coumaric and gallic acids. Neoxanthin, violaxanthin, zeaxanthin, lutein and β-carotene were identified in the extracts of all strains, while canthaxanthin was only identified in Picochlorum sp. SBL2. Taken together, these results strongly suggest that the microalgae included in this work could be used as sources of added-value products that could be used to upgrade the final biomass value.     

Preparation of Chito-Oligomers by Hydrolysis of Chitosan in the Presence of Zeolite as Adsorbent

An increasing interest has recently been shown to use chitin/chitosan oligomers (chito-oligomers) in medicine and food fields because they are not only water-soluble, nontoxic, and biocompatible materials, but they also exhibit numerous biological properties, including antibacterial, antifungal, and antitumor activities, as well as immuno-enhancing effects on animals. Conventional depolymerization methods of chitosan to chito-oligomers are either chemical by acid-hydrolysis under harsh conditions or by enzymatic degradation. In this work, hydrolysis of chitosan to chito-oligomers has been achieved by applying adsorption-separation technique using diluted HCl in the presence of different types of zeolite as adsorbents. The chito-oligomers were retrieved from adsorbents and characterized by differential scanning calorimetry (DSC), liquid chromatography/mass spectroscopy (LC/MS), and ninhydrin test.     

Concurrent vaccination of goats with foot and mouth disease (FMD) and peste des petits ruminants (PPR) booster vaccines

Foot and mouth disease (FMD) remains subclinical and self-limiting in small ruminants, but risk of spread of infection to susceptible cohorts is of great epidemiological significance; therefore, small ruminants must be included in vaccination campaigns in FMD endemic regions. Three groups of goats already immunized against peste des petits ruminants (PPR) were vaccinated with FMD and PPR vaccines alone or concurrently. The specific antibody response against three FMD virus strains and PPR virus were evaluated by competitive enzyme-linked immunosorbent assay (cELISA). Goats concurrently vaccinated with PPR + FMD vaccines had significantly (p < 0.05) higher antibody titers to two serotypes of FMD virus at 28, 45, and 60 days post-immunization compared to goats vaccinated with FMD vaccine alone, while goats vaccinated with PPR vaccines alone or PPR + FMD vaccines concurrently showed similar antibody kinetics against PPR virus up till 60 days post-vaccination. Overall, antibody kinetic curves for all three tested strains of FMD virus and PPR virus were similar in vaccinated groups during the course of experiment.     

Bioactive Hydantoin Alkaloids from the Red Sea Marine Sponge Hemimycale arabica

In the course of our continuing efforts to identify bioactive secondary metabolites from Red Sea marine invertebrates, we have investigated the sponge Hemimycale arabica. The antimicrobial fraction of an organic extract of the sponge afforded two new hydantoin alkaloids, hemimycalins A and B (2 and 3), together with the previously reported compound (Z)-5-(4-hydroxybenzylidene)imidazolidine-2,4-dione (1). The structures of the compounds were determined by extensive 1D and 2D NMR (COSY, HSQC and HMBC) studies and high-resolution mass spectral determinations. Hemimycalins A (2) and B (3) represent the first examples of the natural N-alkylated hydantoins from the sponge Hemimycale arabica. Compounds 1–3 displayed variable antimicrobial activities against E. coli, S. aureus, and C. albicans. In addition, compound 1 displayed moderate antiproliferative activity against the human cervical carcinoma (HeLa) cell line. These findings provide further insight into the chemical diversity as well as the biological activity of this class of compounds.     

Cover Image,Volume 73, Issue 2

The cover image, by Eugênio E. Oliveira et al., is based on the Research Article Pyrethroid resistance is associated with a kdr‐type mutation (L1014F) in the potato tuber moth Tecia solanivora, DOI: 10.1002/ps.4414 . Photo Credit: Edison Torrado‐Leon.

     

Diversity of bacterial pathogens and their antimicrobial resistance profile among commensal rodents in Qatar

Rodents are sources of many zoonotic pathogens that are of public health concern. This study investigated bacterial pathogens and assessed their antimicrobial resistance (AMR) patterns in commensal rodents in Qatar. A total of 148 rodents were captured between August 2019 and February 2020, and blood, ectoparasites, and visceral samples were collected. Gram-negative bacteria were isolated from the intestines, and blood plasma samples were used to detect antibodies against Brucella spp., Chlamydophila abortus, and Coxiella burnetii. PCR assays were performed to detect C. burnetii, Leptospira spp., Rickettsia spp., and Yersinia pestis in rodent tissues and ectoparasite samples. Antimicrobial resistance by the isolated intestinal bacteria was performed using an automated VITEK analyzer. A total of 13 bacterial species were isolated from the intestine samples, namely Acinetobacter baumannii, Aeromonas salmonicida, Citrobacter freundii, Citrobacter koseri, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Hafnia alvei, Klebsiella pneumoniae, Providencia stuartii, Proteus mirabilis, Pseudomonas aeruginosa, and Salmonella enterica. The majority of them were E. coli (54.63%), followed by P. mirabilis (17.59%) and K. pneumoniae (8.33%). Most of the pathogens were isolated from rodents obtained from livestock farms (50.46%), followed by agricultural farms (26.61%) and other sources (22.94%). No antibodies (0/148) were detected against Brucella spp., C. abortus, or C. burnetii. In addition, 31.58% (6/19) of the flea pools and one (1/1) mite pool was positive for Rickettsia spp., and no sample was positive for C. burnetii, Leptospira spp., and Y. pestis by PCR. A total of 43 (38%) bacterial isolates were identified as multidrug resistant (MDR), whereas A. salmonicida (n?=?1) did not show resistance to any tested antimicrobials. Over 50% of bacterial MDR isolates were resistant to ampicillin, cefalotin, doxycycline, nitrofurantoin, and tetracycline. The presence of MDR pathogens was not correlated with rodent species or the location of rodent trapping. Seven (11.86%) E. coli and 2 (22.2%) K. pneumoniae were extended-spectrum beta-lactamases (ESBL) producers. These findings suggest that rodents can be a source of opportunistic bacteria for human and animal transmission in Qatar. Further studies are needed for the molecular characterization of the identified bacteria in this study.

     

In Vitro Evaluation of Liquid‐stored Buffalo Semen at 5°C Diluted in Soya Lecithin Based Extender (Bioxcell®), Tris‐Citric Egg Yolk,Skim Milk and Egg Yolk‐Citrate Extenders

This study was designed to compare the quality of liquid‐stored buffalo bull spermatozoa in soya lecithin based extender Bioxcell^® (BIOX), milk (MILK), tris‐citric egg yolk (TEY) and egg yolk‐citrate (EYC) extender at 5°C. Semen was collected from five Nili‐Ravi buffalo (Bubalus bubalis) bulls of 6–7 years of age with artificial vagina over a period of 3 weeks (two consecutive ejaculates once in a week). Semen ejaculates having more than 60% motility were pooled, split into four aliquots, diluted (37°C; 10 × 10⁶ motile spermatozoa/ml), cooled from 37 to 5°C in 2 h (0.275°C/min) and stored for 5 days. Sperm motility, viability, plasma membrane integrity (PMI) and normal acrosomal ridge were studied at first, third and fifth day of storage. Higher values of progressive sperm motility (%), sperm viability (%), sperm PMI (%) and normal apical ridge (%) were observed in BIOX, MILK and TEY extenders at first, third and fifth day of storage than EYC extender. Progressive sperm motility, sperm viability and sperm PMI in BIOX^® extender were not different from MILK and TEY extenders at 1st and third day storage period. However, at fifth day of storage, the values for these parameters remained significantly higher (p < 0.05) in BIOX^® compared with MILK, TEY and EYC extenders. At fifth day of storage, the semen quality parameters for Bioxcell^® were comparable to those with MILK and TEY extenders at third day of storage. In conclusion, motility, viability and PMI of buffalo bull spermatozoa remained similar in Bioxcell^®, milk and TEY extender at first and third days of storage at 5°C. Yet, the values for the aforementioned parameters in Bioxcell^® were higher compared with milk, TEY and EYC extender at fifth day of storage at 5°C.