Ultrastructure abnormalities of collagen and elastin in Arab patients with arterial tortuosity syndrome.

Arterial tortuosity syndrome (ATS) is a rare autosomal recessive disease characterized by elongation and tortuosity of the large- and medium-sized arteries. ATS patients display features that are also found in Ehlers-Danlos syndrome (EDS) patients. ATS is caused by pathogenic mutations in the SLC2A10 gene, which encodes for the glucose transporter, GLUT10. This study aimed at examining the ultrastructure of skin for abnormalities that can explain the loose skin and arterial phenotypes of Arab patients with the p.S81R mutation in SLC2A10. Forty-eight patients with SLC2A10 mutation were recruited for this study. Skin biopsy specimens from three children with ATS and a healthy child were examined by electron microscopy to determine the ultrastructure of collagen and elastin. Histopathologic staining of sections from tissue biopsy specimens was also performed. Large spaces were observed among the collagen fibrils in the skin biopsy specimens obtained from ATS patients, suggesting disorganization of the collagen structures. Furthermore, elastin fiber contents and their thickness are reduced in the skin. In small muscular arteries in the skin from ATS patients, discontinuous internal elastic lamina, lack of myofilaments, and disorganized medial smooth muscle cells with vacuolated cytoplasm are present. The disorganization of collagen fibrils and reduced elastin contents in the skin may explain the loose skin phenotype of ATS patients similar to the EDS patients. The lack of elastin in small muscular arteries may have contributed to the development of arterial tortuosity in these patients.

Vitamin D Receptor FokI, ApaI, and TaqI Polymorphisms in Lead Exposed Subjects From Saudi Arabia.

Vitamin D receptor (VDR) gene polymorphisms were reported to influence blood lead levels (BLL) and the response of subjects to the symptoms of lead toxicity. However, no studies have been conducted in the Saudi Arabian population which has unique ethnicity and socio-demographic features. This study examined the polymorphisms in exon 2 (allele 1) and intron 8 (allele 2 and allele 3) of VDR gene and their relation to BLLs. As per the CDC guidelines, the recruited lead-exposed workers (N = 130) were categorized to two groups viz., low BLL group (<10 µg/dL) and high BLL group (>10 µg/dL). The low BLL group had a mean BLL of 4.37 µg/dL, while the high BLL group had levels of 18.12 µg/dL (p < 0.001). Overall, the genetic variants, TC and CC in the VDR FokI were significantly associated with a risk of lead toxicity and the allele "C" was a risk factor (p = 0.00026). Furthermore, the TT genotype of VDR ApaI significantly increased the risk of developing lead poisoning (p = 0.0006). The VDR TaqI SNP was not significantly associated with lead toxicity. The highest BLLs for VDR FokI-CC, VDR ApaI-GG, and VDR TaqI-TT genotypes from High BLL group were 18.42, 15.26, and 18.75 µg/dL, respectively. Older age (51-60 years) was found to be a significant confounding factor for BLLs (p = 0.012). Additional studies in larger sample sizes are needed to firmly establish the role of VDR genotypes and genetic susceptibility to lead poisoning.

Evaluation of cytotoxicity and genotoxicity of pesticide mixtures on lymphocytes.

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.

Spectral detection of thalassemia: a preliminary study.

BACKGROUND: Thalassemias (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. RESULTS: In 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%. CONCLUSION: This 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.

Phylogenetic analysis of cubilin (CUBN) gene.

Cubilin, (CUBN; also known as intrinsic factor-cobalamin receptor [Homo sapiens Entrez Pubmed ref NM_001081.3; NG_008967.1; GI: 119606627]), located in the epithelium of intestine and kidney acts as a receptor for intrinsic factor - vitamin B12 complexes. Mutations in CUBN may play a role in autosomal recessive megaloblastic anemia. The current study investigated the possible role of CUBN in evolution using phylogenetic testing. A total of 588 BLAST hits were found for the cubilin query sequence and these hits showed putative conserved domain, CUB superfamily (as on 27(th) Nov 2012). A first-pass phylogenetic tree was constructed to identify the taxa which most often contained the CUBN sequences. Following this, we narrowed down the search by manually deleting sequences which were not CUBN. A repeat phylogenetic analysis of 25 taxa was performed using PhyML, RAxML and TreeDyn softwares to confirm that CUBN is a conserved protein emphasizing its importance as an extracellular domain and being present in proteins mostly known to be involved in development in many chordate taxa but not found in prokaryotes, plants and yeast.. No horizontal gene transfers have been found between different taxa.

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.

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.

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 Gunther's 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 14(th) 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.

Prevalence of hemoglobinopathy disorders in adult patients sent for diagnosis of anemia in saudi arabia.

BACKGROUND: Hemoglobinopathies are a very heterogeneous group of congenital hemolytic anemia. AIM OF THE STUDY: Was to detect the prevalence of hemoglobinopathy disorders in adult patients sent for diagnosis of anemia by high-performance liquid chromatography techniques in Saudi Arabia. SUBJECTS: A total of 329 blood samples from suspected cases with an average age of 39±6.49 of both sexes (140 men and 189 women) were included in the study. MAIN OUTCOME MEASURE: A total of 118 (35.9%) patients were found to have a normal pattern of hemoglobin (Hb)-electrophoresis with a mean level of HbA (α(2)ß(2)) (97.0%±0.40%), HbA2 (α(2)δ(2)) (2.72%±0.55%), and HbF (α(2)γ(2)) (0.59%±0.25%). One hundred twenty (36.5%) subjects were normal with iron deficiency masking the thalassemic trait (Hb: 8.71±1.7 g/dL, iron level: 3.62±2.7 µg/dL, total iron-binding capacity: 86.9±21.5 µg/dL). RESULTS: The remaining 91 (27.66%) patients showed different abnormal hemoglobin variants; 5 (1.5%) subjects had persistence of fetal HbF (7.12%±11.1%). Thirty-three (10.0%) subjects had ß-thalassemia with a high level of HbA2 (>5.54%). Eleven (3.3%) had ß-thalassemic/sickle cell disease. Twenty-nine (8.8%) had sickle cell trait with a level of HbS α(2)ß(S)(2)<45%, and thirteen (4.0%) had sickle cell disease with a high level of HbS ≥45%. CONCLUSION: ß-thalassemia and sickle cell anemia are the most common monogenic disorders in Saudi Arabia. This is a serious health threat to our nation, if it is allowed to continue without taking preventive measures.

An analysis of hematological parameters to assess the prevalence of anemia in elderly subjects from Saudi Arabia.

BACKGROUND: Anemia is one of the most common problems encountered in general practice. Very little is known about the prevalence of anemia in the elderly. AIM OF THE STUDY: To estimate the prevalence of anemia in an elderly population from Saudi Arabia. SETTING: Razi polyclinic Umalhamam, Riyadh. SUBJECTS: A total of 1558 elder subjects were involved in these studies, and 150 control healthy subjects of similar age and gender were included in these studies. MAIN OUTCOME MEASURE: The prevalence of anemia in an elder population of both women and men of Saudi is 12.9%. RESULTS: Commonly used hematological indices were determined in an elder group of subjects over 60 years old. A total of 201 (12.9%) out of 1558 elders were found to have anemia: 154 (9.9%) had moderate anemia, and 47 (3.1%) severe anemia. A higher frequency of anemia was observed in women (18%) with 13.9% subjects having moderate anemia and 4.1% subjects having severe anemia. In men, anemia was observed in 5.06% of patients (moderate [3.87%] and severe [1.31%]). In an analysis comparing moderate and severe anemia, red blood cell indices were found to be significantly higher frequency with a p-value (p<0.001) for all red blood cell indices. CONCLUSION: It can be concluded that anemia in the elderly is an extremely common problem and found more often in women than in men and is associated with increased mortality and poorer health-related quality of life in this group of the population of Saudi Arabia.