Abbas H Alsaeed

Evaluation of cytotoxicity and genotoxicity of pesticide mixtures on lymphocytes

Abstract The cytotoxicity and genotoxicity of pesticide mixtures viz. endosulfan + chlorpyrifos, chlorpyrifos + profenofos, and endosulfan + profenofos were evaluated on cultured human peripheral blood lymphocytes using assays for cell viability, and genotoxicity using chromosomal aberrations test and comet assay. The LC50 values for cytotoxicity were 3.50 μM, 4.18 μM, and 10.5 μM for profenofos, endosulfan, and chlorpyrifos respectively. When combined in equimolar concentrations, the LC50 values for cytotoxicity were 1.4 μM, 1.8 μM, and 2.0 μM for endosulfan + chlorpyrifos, chlorpyrifos + profenofos, and endosulfan + profenofos, respectively. Higher concentrations of individual pesticides (0.5–4.0 μM) but very low concentrations of pesticide mixtures caused significant DNA damage. Additive index values indicated a synergistic effect of toxicity for endosulfan + chlorpyrifos combination (1.12 TTU). The binary mixture of chlorpyrifos + profenofos showed an additive toxicity (0.46 TTU) while an antagonistic effect was observed for endosulfan + profenofos combination. Synergism could be due to these complementary pesticides simultaneously acting in different ways, magnifying their efficacy, whereas an additive interaction would imply that the chemicals are acting by the same mechanism and at the same target. Analysis of toxicity of pesticide mixtures may serve as important biomarker for occupational and household exposure to pesticides, with different modes of action.

Effect of sodium fluoride on neuroimmunological parameters, oxidative stress and antioxidative defenses.

Abstract Aims: This study was designed to evaluate the effect of sodium fluoride (NaF) in inducing neuroimmunological, oxidative and antioxidative damage. Methods: Twenty-four male Wistar rats broadly grouped into four groups containing six rats in each were fed with drinking water containing 20 ppm, 60 ppm, 100 ppm and 0.8 ppm (control) NaF. After 90 days, rats were sacrificed to assess the level of fluoride content and various neurotransmitters in brain. The levels of CD4, natural killer (NK) cells and IgG1 were assessed in blood and spleen. In addition, lipid peroxidation coupled with the levels of various antioxidative enzymes was also recorded. Results: Increase in the NaF concentration resulted in increased fluoride deposition in brain tissue. This increased fluoride content led to increased levels of certain neurotransmitters such as epinephrine, histamine, serotonin and glutamate and decreased levels of norepinephrine, acetylcholine and dopamine in a dose-dependent manner. NaF exposure led to the decrease in the levels of CD4, NK cells and IgG1 coupled with marked increase in lipid peroxidation and impairment of the antioxidative defense system. Conclusion: The result of the study emphasizes the toxic role of high NaF doses on the neurological and immunological functions.

Spectral detection of thalassemia: a preliminary study

BackgroundThalassemias (Thal) are forms of inherited autosomal recessive blood disorders arising out of mutations in the chromosomes 11 or 16. These disorders lead to poor oxygen delivery to blood vessels and consequent splenomegaly, bone deformities, and shorter life spans. The most common detection methods for Thal are complete blood count (CBC) followed by electrophoresis and molecular diagnosis methods, such as high-performance liquid chromatography (HPLC) and polymerase chain reaction (PCR) genotyping. These methods involve sophisticated instrumentations and are cumbersome and expensive.ResultsIn this study an innovative spectral detection method, based on the fluorescence spectra of a set of biomolecules (tyrosine, tryptophan, nicotinamide adenine dinucleotide, and flavin adenine dinucleotide and porphyrins) found in blood components is presented. An algorithm based on the spectral features of such biomolecules of blood components of 20 Thal patients (10 female and 10 male) and 18 age adjusted normal controls (4 female and 14 male) demonstrate reasonable level of classification with sensitivity and specificity values exceeding 90%.ConclusionThis new technique could be of significant value for Thal detection, diagnosis, and subsequent genetic counselling and could be adapted for use in small primary health centres.

Fluorescence spectral diagnosis of malaria – a preliminary study

BackgroundMalaria is the most common disease transmitted by the bite by an infected female anopheles mosquito and caused by the plasmodium parasite. It is mostly prevalent in subtropical regions receiving abundant rain and supporting copious mosquito breeding. This disease is generally detected by the microscopic examination of blood films or antigen based rapid diagnostic test. Only occasionally the parasite DNA is detected using polymerase chain reaction in certain advanced, expensive laboratories.MethodsAn innovative spectral detection method based on the fluorescence spectra of a set of blood plasma biomolecules [tyrosine, tryptophan, nicotinamide adenine dinucleotide (NAD), and flavin adenine dinucleotide (FAD)] and red blood cell (RBC)-associated porphyrin is being evolved by our group.ResultsThe research so far has exhibited sensitivity and specificity values exceeding 90% based on the spectral features of blood components of 14 malaria patients and 20 numbers of age adjusted normal controls. The fluorescent biomolecules go out of proportion when the malarial parasite breaks down the hemoglobin of blood.ConclusionThis technique has the potential to be used as an alternative diagnostic procedure for malaria since the instrumentation involved is portable and inexpensive.Virtual SlidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_182

Reticulocyte Hemoglobin Content and Iron Deficiency: A Retrospective Study in Adults

AIM To evaluate the reticulocyte hemoglobin content (CHr) in a total of 260 adult patients having anemia of chronic disease (ACD), iron deficiency anemia, and chronic renal failure (CRF) who enrolled at the King Khalid University Hospital (Riyadh, Saudi Arabia). RESULTS Results from this study showed that there was a significant correlation between the CHr and hematological parameters, like hemoglobin, hematocrit, mean cell volume, mean cell hemoglobin, and red blood cell distribution. In addition, a significant correlation was seen between the CHr and biochemical parameters for iron status like serum iron, transferrin saturation, and total iron-binding capacity. CONCLUSIONS The CHr, together with a complete blood count, may provide an alternative to the traditional hematological or biochemical panels for the diagnosis of absolute iron deficiency and functional iron deficiency in the case of ACD and the anemia associated with CRF.

Phylogenetic analysis of uroporphyrinogen III synthase (UROS) gene.

The uroporphyrinogen III synthase (UROS) enzyme (also known as hydroxymethylbilane hydrolyase) catalyzes the cyclization of hydroxymethylbilane to uroporphyrinogen III during heme biosynthesis. A deficiency of this enzyme is associated with the very rare Gunthers disease or congenital erythropoietic porphyria, an autosomal recessive inborn error of metabolism. The current study investigated the possible role of UROS (Homo sapiens [EC: 4.2.1.75; 265 aa; 1371 bp mRNA; Entrez Pubmed ref NP_000366.1, NM_000375.2]) in evolution by studying the phylogenetic relationship and divergence of this gene using computational methods. The UROS protein sequences from various taxa were retrieved from GenBank database and were compared using Clustal-W (multiple sequence alignment) with defaults and a first-pass phylogenetic tree was built using neighbor-joining method as in DELTA BLAST 2.2.27+ version. A total of 163 BLAST hits were found for the uroporphyrinogen III synthase query sequence and these hits showed putative conserved domain, HemD superfamily (as on 14th Nov 2012). We then narrowed down the search by manually deleting the proteins which were not UROS sequences and sequences belonging to phyla other than Chordata were deleted. A repeat phylogenetic analysis of 39 taxa was performed using PhyML and TreeDyn software to confirm that UROS is a highly conserved protein with approximately 85% conserved sequences in almost all chordate taxons emphasizing its importance in heme synthesis.

A Phylogenetic analysis of Heparanase (HPSE) gene.

The Current Study aimed to investigate the possible role of Heparanase protein (HPSE-1, [Entrez Pubmed ref|NP_001092010.1|, heparanase isoform 1 preproprotein [Homo sapiens]) in evolution by studying the phylogenetic relationship and divergence of HPSE-1 gene using computational methods. The Human HPSE protein sequences from various species were retrieved from GenBank database and were compared using sequence alignment. Multiple sequence alignment was done using Clustal-W with defaults and phylogenetic trees for the gene were built using neighbor-joining method as in BLAST 2.2.26+ version. A total of 112 BLAST hits were found for the heparanase query sequence and these hits showed putative conserved domain, Glyco_hydro_79n superfamily. We then narrowed down the search by manually deleting the proteins which were not HPSE-1. These sequences were then subjected to phylogenetic analyses using the PhyML and TreeDyn software. Our study indicated that HPSE-1 is a conserved protein in classes Mammalia, Aves, Amphibia, Actinopterygii and Insecta emphasizing its importance in the physiology of cell membranes. Occurrence of this gene in evolution with conserved sites strengthens the role of HPSE-1 gene and helps in better understanding the biochemical processes that may lead to cancer.