Molecular detection and phylogenetic analysis of Leishmania major in stray dogs in Riyadh Province, Saudi Arabia.

Dogs can act as a reservoir of canine leishmaniasis disease, which is caused by Leishmania species. The study aimed to identify and document the genotype of cutaneous leishmaniasis (CL) in the stray dogs in Riyadh Province using kinetoplast DNA (kDNA) as a target gene by using nested polymerase chain reaction (nPCR). This cross-sectional investigation was conducted over the course of two years, from March 2016 to July 2018, in different districts of Riyadh Province, Saudi Arabia. A total of 237 dogs were examined, only 18 of the dogs were suspected clinically of cutaneous leishmaniasis due to the presence of cutaneous nodules and cutaneous lesion. Biopsy tissue collections were performed and DNA was extracted. CSB2XF and CSB1XR primers were used to amplify the Leishmania kDNA regions. The Leishmania species were detected by specific 13Z and LIR primers by applying nested PCR assay. Nine dogs were found to be positive for Leishmania major. The examined dogs were negative for other Leishmania spp. The phylogenetic analysis and blast results of kDNA showed that the 9 isolates L. major is closely related (99.9%) to the L. major isolate CMG_irfan5, accession number HQ727556.1 from human, Pakistan. This is the first molecular study on dog leishmaniasis from Saudi Arabia confirmed that dogs have a L. major infection. Further epidemiological and molecular investigations are required to study domestic and wild canine infections with L. major and other Leishmania spp in endemic and nonendemic areas of Saudi Arabia as part of leishmaniasis control.

Molecular detection and phylogenetic analysis of Leishmania major in stray dogs in Riyadh Province, Saudi Arabia

@#Dogs can act as a reservoir of canine leishmaniasis disease, which is caused by Leishmania species. The study aimed to identify and document the genotype of cutaneous leishmaniasis (CL) in the stray dogs in Riyadh Province using kinetoplast DNA (kDNA) as a target gene by using nested polymerase chain reaction (nPCR). This cross-sectional investigation was conducted over the course of two years, from March 2016 to July 2018, in different districts of Riyadh Province, Saudi Arabia. A total of 237 dogs were examined, only 18 of the dogs were suspected clinically of cutaneous leishmaniasis due to the presence of cutaneous nodules and cutaneous lesion. Biopsy tissue collections were performed and DNA was extracted. CSB2XF and CSB1XR primers were used to amplify the Leishmania kDNA regions. The Leishmania species were detected by specific 13Z and LIR primers by applying nested PCR assay. Nine dogs were found to be positive for Leishmania major. The examined dogs were negative for other Leishmania spp. The phylogenetic analysis and blast results of kDNA showed that the 9 isolates L. major is closely related (99.9%) to the L. major isolate CMG_irfan5, accession number HQ727556.1 from human, Pakistan. This is the first molecular study on dog leishmaniasis from Saudi Arabia confirmed that dogs have a L. major infection. Further epidemiological and molecular investigations are required to study domestic and wild canine infections with L. major and other Leishmania spp in endemic and nonendemic areas of Saudi Arabia as part of leishmaniasis control

A label-free, PCR-free and signal-on electrochemical DNA biosensor for Leishmania major based on gold nanoleaves.

Detection of leishmaniasis is important in clinical diagnoses. In the present study, identification of Leishmania parasites was performed by a label-free, PCR-free and signal-on ultrasensitive electrochemical DNA biosensor. Gold nanoleaves were firstly electrodeposited by an electrodeposition method using spermidine as a shape directing agent. The biosensor was fabricated by immobilization of a Leishmania major specific DNA probe onto gold nanoleaves, and methylene blue was employed as a marker. Hybridization of the complementary single stranded DNA sequence with the biosensor under the selected conditions was then investigated. The biosensor could detect a synthetic DNA target in a range of 1.0×10-10 to 1.0×10-19molL-1 with a limit of detection of 1.8×10-20molL-1, and genomic DNA in a range of 0.5-20ngµL-1 with a limit of detection of 0.07ngµL-1. The biosensor could distinguish Leishmania major from a non-complementary-sequence oligonucleotide and the tropica species with a high selectivity. The biosensor was applicable to detect Leishmania major in patient samples.

Synthesis and Applications of Nanoflowers.

BACKGROUND: Recent advances in nanotechnology make novel shapes of nanostructured materials with novel physicochemical properties. Different kinds of materials including carbon, metals, alloys, metal oxides, conducting polymers, metal chalcogenides, pnictides and fluorides have been synthesized with small size of <100 nm with shapes resembled to the natural flowers. The objective of this review is to provide a broad overview of the synthesis strategies, effects of different parameters on the morphology of nanoflowers, and their applications. METHODS: A comprehensive search to assess the current evidence for the synthesis routs of nanoflowers and applications was conducted. 487 studies became selected and the quality of papers were appraised and categorized according to type of nanomaterials. Within each section, the nanomaterials addressed specifically. In addition, recent patents were reviewed in a separate section. RESULTS: The nanoflowers exhibited unique properties which were utilized in the design of efficient applications. These nanostructures can be processed with different methods. The configuration of flower-like nanostructures can be controlled by altering experimental parameters, such as the precursor`s ratio, temperature and reaction time. Despite the huge efforts to control and understand the growth mechanism of these nanostructures, some fundamental phenomena are still not well understood. Investigation of the fundamental effects of size and morphology on their properties is required in order to better apply these nanostructured materials. CONCLUSION: The nanoflowers with interesting properties can be used in the design of future devices with various applications. The existence of different routes to synthesis nanoflowers and their unique properties confirm the importance promoted awareness of potential benefits of nanoflowers in different applications. The reviewed patents emphasized the importance of these nanostructures. Therefore, research efforts based on nanoflowers are dynamic and applicable in various fields.

An ultrasensitive electrochemical genosensor for Brucella based on palladium nanoparticles.

Palladium nanoparticles were potentiostatically electrodeposited on a gold surface at a highly negative potential. The nanostructure, as a transducer, was utilized to immobilize a Brucella-specific probe and the process of immobilization and hybridization was detected by electrochemical methods. The proposed method for detection of the complementary sequence and a non-complementary sequence was applied. The fabricated genosensor was evaluated for the assay of the bacteria in the cultured and human samples with and without PCR. The genosensor could detect the complementary sequence with a sensitivity of 0.02 µA dm(3) mol(-1), a linear concentration range of 1.0 × 10(-12) to 1.0 × 10(-19) mol dm(-3), and a detection limit of 2.7 × 10(-20) mol dm(-3).

Label-free electrochemical aptasensing of the human prostate-specific antigen using gold nanospears.

Gold nanospears were electrodeposited with the assistance of arginine as a soft template and precise selection of experimental parameters. The nanospears were then employed as a transducer to immobilize an aptamer of prostate-specific antigen (PSA) and fabrication of a label-free electrochemical aptasensor. The aptasensor was employed for the detection of PSA with a linear concentration range of 0.125-200ngmL(-1) and a limit of detection of 50pgmL(-1). The aptasensor was successfully applied to detect PSA in blood serum samples of healthy and patient persons.

Nanostructured materials in electroanalysis of pharmaceuticals.

Basic strategies and recent developments for the enhancement of the sensory performance of nanostructures in the electroanalysis of pharmaceuticals are reviewed. A discussion of the properties of nanostructures and their application as modified electrodes for drug assays is presented. The electrocatalytic effect of nanostructured materials and their application in determining low levels of drugs in pharmaceutical forms and biofluids are discussed.

Marked benefits in physical activity and well-being, but not in functioning domains, 2 years after successful epilepsy surgery in children.

In this first study comparing epilepsy-specific quality-of-life measures of children after epilepsy surgery (2.4 years after focal resection) with those of a matched comparison group of nonoperated patients, seizure severity, medication side effects, overall quality of life, general health, physical activity, and well-being were better in surgical patients (70.6% seizure free vs 8.3%). Cognitive, social, and behavioral functioning did not differ, suggesting that these may require additional interventions during postsurgical follow-up.

Risk factors for development of subclinical hypothyroidism during valproic acid therapy.

OBJECTIVE: To identify risk factors for subclinical hypothyroidism (SCH) (thyroid-stimulating hormone levels >5 mIU/mL) in patients receiving valproate (VPA) therapy. STUDY DESIGN: During a period of 2 years, consecutive patients with epilepsy receiving VPA and a control group of patients with diseases other than epilepsy attending a tertiary care neurology clinic were screened for SCH. The 2 groups were compared. The association between SCH and specific risk factors was investigated with bivariate and multivariate analyses. RESULTS: Thirty-six of 143 patients receiving VPA (25.2%, mean age +/- SD: 8.5 +/- 6.6 years) and none of the 35 control subjects had SCH (P < .001). Predictors of SCH were younger age (OR: 1.15, cutoff age 3.9 years); duration of treatment between 6 and 24 months versus <6 months (OR: 2.98) and >24 months (OR: 2.66); VPA polytherapy with enzyme-inducing agents (OR: 6.08), or polytherapy with non-enzyme-inducing agents (OR: 3.34) compared with VPA monotherapy. Most (88.2%) patients with duration of therapy >2 years were older than 3.9 years. CONCLUSION: Risk factors for SCH were young age, co-medication with antiepileptic drugs, and duration of therapy between 6 and 24 months. Screening patients with these risk factors may be warranted.

Comparison of carotid Doppler ultrasound and computerised tomographic angiography in the evaluation of carotid artery stenosis.

PURPOSE: To compare results of carotid Doppler ultrasound (CDUS) and spiral computerised tomographic angiography (CTA) in patients with suspected carotid artery stenosis and to evaluate their combined effect on decision making for carotid endarterectomy (CEA). METHODS: A total of 107 patients were studied. All of the patients had CDUS followed by CTA as a standard method of investigation. Data included the indications for investigation, stenosis degree measured in both modalities, in addition to difficulties and limitations faced while doing them. RESULTS: Out of the 214 carotid scans performed, 187 scans were included in the comparison, while 27 scans were excluded due to inadequate data or imaging difficulties. The overall concordance between both CDUS and CTA was 79.1% (148/187) (95% CI 0.72-0.83). CDUS under-estimated and over-estimated the degree of stenosis in 26/187 (14%, 95% CI 0.09-0.19) and 13/187 (7%, 95% CI 0.04-0.12), respectively. When CTA was considered in conjunction with CDUS, the decision regarding operative treatment was changed in 29/187 cases (16%) (95% CI 0.11-0.21). CONCLUSIONS: CDUS remains the first line non-invasive imaging for carotid artery stenosis. However, in cases where it is inconclusive, CTA is an excellent, reliable, minimally invasive, and outpatient alternative for patient selection for CEA.

Two randomized vitamin D trials in ambulatory patients on anticonvulsants: impact on bone.

OBJECTIVE: To investigate the effects of two doses of vitamin D given over 1 year on bone density in ambulatory patients on long-term antiepileptic drug (AED) therapy. METHODS: We conducted two parallel, randomized, controlled trials in 72 adults (18 to 54 years old) and 78 children and adolescents (10 to 18 years) on long-term AED therapy. They received either low-dose vitamin D 400 IU/day or high-dose vitamin D 4,000 IU/day (adults) and 2,000 IU/day (children/adolescents). Bone mineral density (BMD) was measured using dual-energy x-ray absorptiometry. RESULTS: In adults, baseline BMD was lower than that of age- and gender-matched controls vs either a Western or an ethnically identical population. After 1 year, there were significant increases in BMD at all skeletal sites compared to baseline in the high-, but not in the low-dose treatment group. However, BMD at 1 year remained below normal. In children, baseline BMD was normal vs age- and gender-matched controls and showed significant and comparable increases in both treatment groups. CONCLUSIONS: In ambulatory adults on antiepileptic drugs, high-dose vitamin D therapy substantially increased bone mineral density at several skeletal sites. In children, both doses resulted in comparable increases in bone mass.

Genotyping human platelet alloantigens (HPA 1-5) in Saudis from Eastern Province, Saudi Arabia.

In this study we report for the first time the distribution of human platelet alloantigens (HPA) in Saudis. These antigens are implicated in the pathophysiology of alloimmune thrombocytopenia. We collected blood samples from 84 healthy male Saudi blood donors. DNA isolated by salting-out and ethanol precipitation was amplified for genes HPA 1-5 using the polymerase chain reaction/sequence specific primer method. We found high HPA-1 polymorphism similar to Caucasians. HPA-4 polymorphism in Saudis was, however, greater than in Caucasians, and more similar to that of the Japanese. These results suggest that both these two HPA systems may be clinically important in Saudis.

Genotyping human platelet alloantigens [HPA 1-5] in Saudis from Eastern Province, Saudi Arabia

In this study we report for the first time the distribution of human platelet alloantigens [HPA] in Saudis. These antigens are implicated in the pathophysiology of alloimmune thrombocytopenia. We collected blood samples from 84 healthy male Saudi blood donors. DNA isolated by salting-out and ethanol precipitation was amplified for genes HPA 1-5 using the polymerase chain reaction/sequence specific primer method. We found high HPA-1 polymorphism similar to Caucasians. HPA-4 polymorphism in Saudis was, however, greater than in Caucasians, and more similar to that of the Japanese. These results suggest that both these two HPA systems may be clinically important in Saudis

Immunophenotypic pattern of childhood acute lymphoblastic leukemia.

Full text is available as a scanned copy of the original print version.

Primary antibody deficiency in Arabs: first report from eastern Saudi Arabia.

Epidemiological studies have shown wide geographical and racial variations in the prevalence and pattern of immunodeficiency diseases. As there is no national registry, very little is known of the prevalence and nature of humoral immunodeficiency in the Arabian peninsula. We report here for the first time the analysis of serum immunoglobulin (Ig) levels in 2000 consecutive patients (age, 1-80 years). They were seen over a period of 6 years and were referred to us from six district hospitals for suspected immunodeficiency, autoimmunity, allergy, or immunoglobulin dyscrasia. Forty-six were found to be immunodeficient, in whom at least one of the Ig class was low; 15 had secondary immunodeficiency. The remaining 31 cases, representing 1.5% of the population studied (giving a prevalence of 1550/100,000 hospital registered patients), were categorized into four primary humoral immunodeficiency groups: these included, in order of frequency, (i) selective IgA deficiency (45%; 700/100,000) (ii) common variable immunodeficiency (CVID) (29%; 450/100,000), (iii) agammaglobulinemia (16%; 250/100,000), and (iv) selective IgG deficiency (10%; 150/100,000). Compared with similar hospital-based surveys in the west the prevalence of humoral immunodeficiency seems to be higher in Arabs; this in part may be related to race and higher rate of consanguinity. Most patients with IgA deficiency had either infection, atopy or autoimmunity. Compared with some other races, agammaglobulinemia (X- and non-X-linked) seems to be more prevalent.

Common variable immunodeficiency with CD4+ T lymphocytopenia and overproduction of soluble IL-2 receptor associated with Turner's syndrome and dorsal kyphoscoliosis.

An unusual combination of common variable immunodeficiency (CVID) and Turner's syndrome in a Saudi woman aged 20 years is presented. In addition to panhypogammaglobulinaemia, the patient had CD4+ T lymphocytopenia; however, there was evidence of in vivo activation of T cells and overproduction of soluble interleukin 2 receptor in culture supernate. Mantoux test was positive, but lymphoblastic response to non-specific mitogen was impaired. Immunogenetically the patient was HLA-DR3 positive and karyotypically she was a mosaic (45XO/46XX) with ring X chromosome (46Xr(X)). The presence of severe kyphoscoliosis was possibly related to ring X chromosome. This case highlights the grave consequences of the delayed diagnosis of immunodeficiency and emphasises the heterogeneous nature of CVID.

Wessely-type immune ring following phototherapeutic keratectomy.

Immune rings following photorefractive keratectomy (PRK) have been reported but have not been described in detail. This case report describes an immune ring after phototherapeutic keratectomy (PTK) in a patient with long-standing superficial corneal scars. A dense white ring formed in the peripheral cornea on the fourth day following surgery. The patient was treated with antibiotics until negative cultures were reported 48 hours later. A biopsy was taken and examined by light microscopy using hematoxylin-eosin and Mason's trichrome staining. The stroma showed focal keratocyte depopulation with nuclear fragments, occasional polymorphonuclear leucocytes, and an active fibroblastic reaction. No lymphocytes or plasma cells were seen. Clinically, the immune ring faded slowly and was still apparent 9 months after the PTK. Studies of similar cases are required to clarify the mechanisms responsible for this phenomenon.