Sooad Al-Daihan

Measurement of selected ions related to oxidative stress and energy metabolism in Saudi autistic children

OBJECTIVES Autism is a developmental disorder characterized by social and emotional deficits, language impairments and stereotyped behaviors that manifest in early postnatal life. This study aims to clarify the role of selected ions related to energy metabolism as a consequence of oxidative stress in the deterioration accompanied autism. MATERIALS AND METHODS Malonaldehyde as measure of lipid peroxidation, Na(+)/K(+) ion pump (ATPase), together with the concentrations of Na(+), K(+), Mg(2+), Ca(2+) and Pb(2+) were determined in plasma of 30 Saudi autistic patients and compared to 30 age-matching control samples. RESULTS The obtained data recorded that Saudi autistic patients have a remarkable higher activities of Na(+)/K(+) ATPase and high levels of lipid peroxidation compared to control. In addition, they have significantly elevated levels of K(+) and Pb(2+) while Ca(2+) recorded a significantly lower level compared to age-matching control subjects. On the other hand both Mg(2+) and Na(+) were non-significantly changed in autistic patients. CONCLUSION Alteration of the selected measured ions confirms that oxidative stress and defective mitochondrial energy production could represent the primary causative factor in the pathogenesis of autism.

On the protective effect of omega-3 against propionic acid-induced neurotoxicity in rat pups

BackgroundsThe investigation of the environmental contribution for developmental neurotoxicity is very important. Many environmental chemical exposures are now thought to contribute to the development of neurological disorders, especially in children. Results from animal studies may guide investigations of human populations toward identifying environmental contaminants and drugs that produce or protect from neurotoxicity and may help in the treatment of neurodevelopmental disorders.ObjectiveTo study the protective effects of omega-3 polyunsaturated fatty acid on brain intoxication induced by propionic acid (PPA) in rats.Methods24 young male Western Albino rats were enrolled in the present study. They were grouped into three equal groups; oral buffered PPA-treated group given a nuerotoxic dose of 250 mg/Kg body weight/day for 3 days; omega-3 - protected group given a dose of 100 mg/kg body weight/day omega-3 orally daily for 5 days followed by PPA for 3 days, and a third group as control given only phosphate buffered saline. Tumor necrosis factor-α, caspase-3, interlukin-6, gamma amino-buteric acid (GABA), serotonin, dopamine and phospholipids were then assayed in the rats brains tissue of different groups.ResultsThe obtained data showed that PPA caused multiple signs of brain toxicity as measured by depletion of gamaaminobyteric acid (GABA), serotonin (5HT) and dopamine (DA) as three important neurotransmitters that reflect brain function. A high significant increase of interlukin-6 (Il-6), tumor necrosis factor-α (TNF-α) as excellent markers of proinflammation and caspase-3 as a proapotic marker were remarkably elevated in the intoxicated group of rats. Moreover, brain phospholipid profile was impaired in PPA-treated young rats recording lower levels of phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylcholine (PC).ConclusionsOmega-3 fatty acids showed a protective effects on PPA - induced changes in rats as there was a remarkable amelioration of most of the measured parameters (i.e. higher GABA, 5HT, DA, PE, PS and PC) and lower Il-6, TNF-α and caspase-3.

Antibacterial activities of extracts of leaf, fruit, seed and bark of Phoenix dactylifera

The antibacterial activities of different parts of local Phoenix dactylifera were investigated in vitro. Dried leaf, fruit, seed and tree bark were extracted with water, methanol and acetone. Antibacterial property of the extracts was evaluated against Staphylococcus aureus , Streptococcus pyogenes, Escherichia coli and Pseudomonas aeruginosa using the disc diffusion method. Overall analysis of the antibacterial activity of various extracts revealed that the highest inhibitory activity was produced by the fruit extract (18.2 ± 0.55 mm) as compared to the leaf, bark and seed extracts. All the extracts from the different parts of the plant showed antibacterial activity against most tested microorganisms. On the whole, aqueous extracts have the least antibacterial activity as compared to methanol and acetone extracts. The antibacterial activity against the Gram-positive strains was the highest in the acetone fruit extract against S. aureus (18.2 ± 0.55 mm). The most active extract against Gram-negative bacteria was methanol extract from the leaves with a 13.5 ± 0.33 mm inhibition zone for E. coli followed by 12.5 ± 0.88 mm for P. aeruginosa. Phytochemical analysis revealed the presence of carbohydrates and alkaloids in all parts, and flavonoids, steroids, saponins and tannins were present in some parts. Key words : Antibacterial activity, Phoenix dactylifera, disc diffusion assay, extracts, inhibition zone.

Phytochemical constituents and antibacterial activity of some green leafy vegetables

OBJECTIVE To investigate the antibacterial activity and photochemicals of five green leafy vegetables against a panel of five bacteria strains. METHODS Disc diffusion method was used to determine the antibacterial activity, while kanamycin was used as a reference antibiotic. The phytochemical screening of the extracts was performed using standard methods. RESULTS All methanol extracts were found active against all the test bacterial strains. Overall maximum extracts shows antibacterial activity which range from 6 to 15 mm. Proteins and carbohydrates was found in all the green leaves, whereas alkaloid, steroids, saponins, flavonoids, tannins were found in most of the test samples. CONCLUSIONS The obtain result suggests that green leafy vegetables have moderate antibacterial activity and contain various pharmacologically active compounds and thus provide the scientific basis for the traditional uses of the studied vegetables in the treatment of bacterial infections.

Purification, characterization and bactericidal activities of phospholipase A2 from the dromedary intestine

We purified a protein from the dromedary small intestine that displayed potent bactericidal activity against Gram-positive bacteria, and identified it as group-IIA phospholipase A₂ (DrPLA₂-IIA) by NH₂-terminal sequencing and enzymatic measurements. In fact, our findings revealed that the purified PLA₂-IIA was a monomeric protein with a molecular mass of about 14 kDa. Pure enzyme has a specific activity of 329 ± 25 U/mg at optimal conditions (pH 9.5 and 45 °C) in the presence of 6 mM NaDC and 7 mM CaCl₂ with egg yolk emulsion as substrate and binds with a higher affinity to PE than PS and PC. Furthermore, the DrPLA2-IIA activity was dependent on Ca(2+); other cations (Cd(2+), Co(2+), Fe(2+), Mg(2+), Mn(2+), and Zn(2+)) reduced the enzymatic activity notably, suggesting that the arrangement of the catalytic site presents an exclusive structure for Ca(2+). On the other hand, DrPLA2-IIA was highly bactericidal against Gram-positive bacteria with inhibition zones and IC₅₀ values in the range of 21-27 mm and 3.7-8 μg/ml, respectively, whereas Gram-negative bacteria exhibited a much higher resistance. These observations suggest that the main physiological role of DrPLA2-IIA could be the defense of the intestine against bacterial infections.

Activities of key glycolytic enzymes in the plasma of Saudi autistic patients

Objective: Measurement of plasma levels of lactate, lactate oxidase (LOX), pyruvate kinase (PK), and hexokinase (HK) as possible glycolytic parameters to assess brain damage in autistic patients. Design and methods: Plasmatic levels of lactate, LOX, PK, and HK were determined in 20 autistic children aged 3–15 years and 20 age-matching healthy control subjects. Results: Plasmatic levels of lactate and LOX were significantly higher in autistic patients compared to healthy subjects and that of PK and HK were significantly lower in these patients as compared to controls. This could reflect the impaired metabolism of astrocytes, the brain cells responsible for the production and provision of lactate, as the primary metabolic fuel for neurons. Conclusion: Remarkably different levels of plasma glycolytic parameters were recorded in Saudi autistic patients. This could be correlated to the impairment of energy metabolism, glutathione depletion, and lead intoxication previously detected in the same investigated samples. The identification of biochemical markers related to autism would be advantageous for earlier clinical diagnosis and intervention.

Potency of pre–post treatment of coenzyme Q10 and melatonin supplement in ameliorating the impaired fatty acid profile in rodent model of autism

Background Abnormalities in fatty acid metabolism and membrane fatty acid composition play a part in a wide range of neurodevelopmental and psychiatric disorders. Altered fatty acid homeostasis as a result of insufficient dietary supplementation, genetic defects, the function of enzymes involved in their metabolism, or mitochondrial dysfunction contributes to the development of autism. Objective This study evaluates the association of altered brain lipid composition and neurotoxicity related to autism spectrum disorders in propionic acid (PA)–treated rats. Design Forty-eight young male western albino rats were used in this study. They were grouped into six equal groups with eight rats in each. The first group received only phosphate buffered saline (control group). The second group received a neurotoxic dose of buffered PA (250 mg/kg body weight/day for 3 consecutive days). The third and fourth groups were intoxicated with PA as described above followed by treatment with either coenzyme Q (4.5 mg/kg body weight) or melatonin (10 mg/kg body weight) for 1 week (therapeutically treated groups). The fifth and sixth groups were administered both compounds for 1 week prior to PA (protected groups). Methyl esters of fatty acid were extracted with hexane, and the fatty acid composition of the extract was analyzed on a gas chromatography. Results The obtained data proved that fatty acids are altered in brain tissue of PA-treated rats. All saturated fatty acids were increased while all unsaturated fatty acids were significantly decreased in the PA-treated group and relatively ameliorated in the pre–post melatonin and coenzyme Q groups. Conclusions Melatonin and coenzyme Q were effective in restoring normal level of most of the impaired fatty acids in PA-intoxicated rats which could help suggest both as supplements to ameliorate the autistic features induced in rat pups.

Genetic polymorphism and expression of HSF1 gene is significantly associated with breast cancer in Saudi females

The transcription factor, heat shock factor 1 (HSF1), influences the expression of heat shock proteins as well as other activities like the induction of tumor suppressor genes, signal transduction pathway, and glucose metabolism. We hypothesized that single nucleotide polymorphisms (SNPs) in HSF1 gene might affect its expression or function which might have an influence on the development of breast cancer. The study group included 242 individuals (146 breast cancer patients and 96 healthy controls). From the cancer patients, genomic DNA was extracted from 96 blood samples and 50 Formalin-Fixed Paraffin Embedded (FFPE) tissues, while from the controls DNA were extracted from blood only. Genotype was carried out for four SNPs in the HSF1 gene (rs78202224, rs35253356, rs4977219 and rs34404564) using Taqman genotyping assay method. The HSF1 expression was investigated using immunohistochemistry on FFPE tissues (cancer tissue and adjacent normal tissue). The SNP rs78202224 (G>T) was significantly associated with increased risk of breast cancer. The combined TT + GT genotype (OR: 6.91; p: 0.035) and the T allele showed high risk (OR: 5.81; p:0.0085) for breast cancer development. The SNP rs34404564 (A>G) had a protective effect against the development of breast cancer. The genotype AG (OR: 0.41; p = 0.0059) and GG+AG (OR: 0.52; p: 0.026) occurred at a significantly lower frequency in the breast cancer patients compared to the frequency in healthy controls. No significant relationship was identified between either rs35253356 (A>G) or rs4977219 (A>C) and breast cancer in Saudi. The HSF1 protein expression was higher in all invasive and in situ breast carcinoma compared to the normal tissue. A stronger positive staining for HSF1 was found in the nucleus compared to the cytoplasm. Our results show that HSF1 gene expression is elevated in breast cancer tissue and two of the studied SNPs correlate significantly with cancer development.

The expression of telomere-related proteins and DNA damage response and their association with telomere length in colorectal cancer in Saudi patients

Background Colorectal cancer is the leading cause of cancer-related deaths in Saudi Arabia. Cancer has a multifactorial nature and can be described as a disease of altered gene expression. The profiling of gene expression has been used to identify cancer subtypes and to predict patients’ responsiveness. Telomere-associated proteins that regulate telomere biology are essential molecules in cancer development. Thus, the present study examined their contributions to colorectal cancer progression in Saudi patients. Methods The expression of hTERT, TRF1, TRF2, POT1, ATR, ATM, Chk1 and Chk2 were measured via real-time PCR in matched cancerous and adjacent tissues of CRC patients. The protein level of hTERT, TRF1, TRF2, ATR, ATM, Chk1 and Chk2 were measured using immunohistochemistry. A region of hTERT core promoter was sequenced via Sanger sequencing. Methylation of CTCF binding site was examined via methylation-specific PCR. Finally, the length of telomere was estimated using q-PCR. Results Our results showed that POT1, ATR, Chk1 and Chk2 show increased expression in CRC relative to the adjacent mucosa. The expression levels of each gene were associated with clinicopathological characteristics of patients with CRC. There was a positive correlation between the age of the patients and hTERT expression. Regarding tumor site, telomere length, ATR, ATM and Chk1 were shown to be altered. No somatic mutation was detected in hTERT core promoter, and no differences in methylation patterns at CTCF binding site in the promoter between normal and cancer tissues. Conclusion Analysis of targeted genes expression in colorectal cancer based on the clinical variables revealed that tumor location and age could have a role in gene expression and telomere length variations and this could be taken under consideration during CRC diagnosis and therapy. Other epigenetic mechanisms could influence hTERT expression in cancers. Our findings warrant further validation through experiments involving a larger number of patients.

High-fat diet stimulates the gut pathogenic microbiota and maintains hepatic injury in antibiotic-treated rats

The gut and the liver are closely linked to each other, as changes in the gut microbiota can play a significant role in the development of many liver diseases. Gut bacteria respond rapidly to changes in diet and thus can affect the liver through their metabolites. The impact of a high lipid diet on the liver in the presence of an altered gut flora modulated by ampicillin was investigated. The study was performed on 30 male Western albino rats randomly divided into 3 groups: control (phosphate buffered saline treated), group II (ampicillin 50 mg/kg for three weeks to induce microbiota alterations and fed on standard diet) and group III (same dose of ampicillin and fed on a lipid rich diet). Stool samples were collected for qualitative determination of bacteria. Serum hepato-specific markers, in addition to Glutathione (GSH), Lipid peroxidase (MDA), Glutathione-S- transferase(GST), and vitamin C in liver tissues, were measured. Altered gut microbiota significantly increased the level of the hepato-specific marker MDA and reduced the GST, GSH and vitamin C levels. However, animals fed a lipid rich diet displayed a more significant shift in hepatic markers and antioxidants. Moreover, a new switch in composition of the gut bacteria was observed by feeding the lipid rich diet. Our study showed that bacterial overgrowth in the gut can be associated with liver dysfunction and that a high lipid diet can promote the overgrowth of some liver damaging microflora during antibiotic treatment.