Publications by authors named "Said Hassan"

34 Publications

Phylogenetic analysis of the 5' untranslated region of HCV from cirrhotic patients in Khyber Pakhtunkhwa, Pakistan.

Sci Rep 2021 07 22;11(1):15023. Epub 2021 Jul 22.

Department of Microbiology and Biotechnology, Abasyn University Peshawar, Khyber Pakhtunkhwa, Pakistan.

Hepatitis C virus (HCV), a small, single-stranded RNA virus with a 9.6 kb genome, is one of the most common causes of liver diseases. Sequencing of the 5' untranslated region (UTR) is usually used for HCV genotyping, but it is less important in numerous subtypes due to its scarce sequence variations. This study aimed to identify genotypes using the 5' UTR of HCV from cirrhotic patients of Khyber Pakhtunkhwa (KP). Serum RNA samples (44) were screened by real time PCR to determine the HCV viral load. Nested PCR was performed to identify cDNA and the 5' UTR. The HCV 5' UTR was sequenced using the Sanger method. MEGA-7 software was used to analyze evolutionary relatedness. After 5' UTR sequencing, 26 samples (59%) were identified as genotype 3, and 2 samples (6%) were identified as genotypes 1, 2 and 4. The most predominant genotype was 3a, and genotype 4 was rarely reported in the phylogenetic tree. Analysis of the HCV 5' UTR is an efficient alternative method for confirmation of various genotypes. Phylogenetic analysis showed that genotype 3 was dominant in the area of KP, Pakistan.
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http://dx.doi.org/10.1038/s41598-021-94063-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8298656PMC
July 2021

Microfabricated potentiometric sensor for personalized methacholine challenge tests during the COVID-19 pandemic.

Biosens Bioelectron 2021 Oct 12;190:113439. Epub 2021 Jun 12.

Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr-El Aini Street, Cairo, 11562, Egypt.

The methacholine challenge test is considered to be the gold standard bronchoprovocation test used to diagnose asthma, and this test is always performed in pulmonary function labs or doctors' offices. Methacholine (MCH) acts by inducing airway tightening/bronchoconstriction, and more importantly, MCH is hydrolyzed by cholinesterase enzyme (ChE). Recently, the American Thoracic Society raised concerns about pulmonary function testing during the COVID-19 pandemic due to recently reported correlation between cholinesterase and COVID-19 pneumonia severity/mortality, and it was shown that cholinesterase levels are reduced in the acute phase of severe COVID-19 pneumonia. This work describes the microfabrication of potentiometric sensors using copper as the substrate and chemically polymerized graphene nanocomposites as the transducing layer for tracking the kinetics of MCH enzymatic degradation in real blood samples. The in-vitro estimation of the characteristic parameters of the MCH metabolism [Michaelis-Menten constant (K) and reaction velocity (V)] were found to be 241.041 μM and 56.8 μM/min, respectively. The proposed sensor is designed to be used as a companion diagnostic device that can (i) answer questions about patient eligibility to perform methacholine challenge tests, (ii) individualize/personalize medical dosing of methacholine, (iii) provide portable and inexpensive devices allowing automated readouts without the need for operator intervention (iv) recommend therapeutic interventions including intensive care during early stages and reflecting the disease state of COVID-19 pneumonia. We hope that this methacholine electrochemical sensor will help in assaying ChE activity in a "timely" manner and predict the severity and prognosis of COVID-19 to improve treatment outcomes and decrease mortality.
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http://dx.doi.org/10.1016/j.bios.2021.113439DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8197613PMC
October 2021

Quality control of dietary supplements: An economic green spectrofluorimetric assay of Raspberry ketone and its application to weight variation testing.

Spectrochim Acta A Mol Biomol Spectrosc 2021 Nov 29;261:120032. Epub 2021 May 29.

Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.

Although Dietary supplements are readily accessible and extensively used worldwide, they are inadequately regulated and consumers are victims of manufacturers' fraud. Thus, quality regulations are required to ensure safety of products available to the public. We propose the first native spectrofluorimetric quality control assay of raspberry ketone, a popular dietary supplement ingredient for weight loss. This work relies on the constant wavelength synchronous scan of the Raspberry Ketone native fluorescence, overcoming the demerits of conventional excitation/ emission spectra. For the best measurement conditions, several parameters were optimized including Δλ value, diluting solvent, medium pH and the effect of surfactants/ macromolecules. In aqueous medium (Δλ = 110 nm), a linear relationship exists between synchronous fluorescence intensity at peak maximum 405.6 nm and solution concentration in the range 300-1500 ng/mL. Method sensitivity was recorded with LOD and LOQ values 60.63 and 183.72 ng/mL; respectively. Validation was done in accordance to International Conference on Harmonization (ICH) guidelines. This simple procedure was successfully applied to the analysis of Raspberry Ketone in commercially available dietary supplement capsules with average recovery 98.67% ± 1.74 and further extended to weight variation testing following the official United States Pharmacopeial (USP) guidelines. Finally, green assessment was done using the ''Analytical Eco-scale'' tool. The total score was 89/100 points revealing excellent greenness of our proposal. Our proposal is simple, eco-friendly and cheap. It can be conveniently adopted for routine quality control practices especially in developing countries.
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http://dx.doi.org/10.1016/j.saa.2021.120032DOI Listing
November 2021

Optimization of localized surface plasmon resonance hot spots in surface-enhanced infrared absorption spectroscopy aluminum substrate as an optical sensor coupled to chemometric tools for the purity assay of quinary mixtures.

Mikrochim Acta 2021 05 22;188(6):195. Epub 2021 May 22.

Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, El-Kasr El-Aini Street, Cairo, ET-11562, Egypt.

Surface-enhanced infrared absorption spectroscopy offers an alternative to conventional IR spectroscopy and utilizes the signal enhancement exerted by the plasmon resonance of nanostructured metal thin films. Citrate-capped silver nanoparticles were prepared in a single-step method, and their morphology was identified using transmission electron microscopy, scanning electron microscopy, ultraviolet/visible spectrophotometry, and Zetasizer. The nanoparticles generated were deposited on the surface of cheap aluminum slides for different durations aiming for the selection of the best time producing a thin film, suitable to act as a lab-on-a-chip SEIRA substrate. These substrates were coupled to partial least squares regression tools for simultaneous resolving of the quinary mixture in commercial dosage forms of bisoprolol, perindopril, bisoprolol acid degradation product, bisoprolol alkali degradation product, and perindoprilat in concentration ranges of 15-75, 60-300, 15-55, 12-60, and 20-80 μg/mL with limits of detection values of 0.69, 3.43, 0.97, 1.25, and 1.09 μg/mL, respectively. Overall, we could demostrate that the localized surface plasmon resonance sensor coupled to chemometrics provides cheap, simple, selective, multiplex, rapid, and molecular specific procedures for impurity detection, which would be beneficial in many applications for quality control and quality accuracy of active pharmaceutical ingredients.
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http://dx.doi.org/10.1007/s00604-021-04845-7DOI Listing
May 2021

Effect of the Prenatal Exposure of Khat on the Skeleton of Developing Rabbit Embryo: Morphometric and Gross Anatomical Study.

Fetal Pediatr Pathol 2020 Oct 7:1-15. Epub 2020 Oct 7.

Department of Anatomy and Embryology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia Egypt.

Background: Khat leaves are chewed by many people worldwide, mainly in Africa and the southwest Arabian peninsula. 27 apparently healthy adult New Zealand rabbits (3 males & 24 females) of 2.5 ± 0.5 kg body weight were used in this work. The animals were divided into 4 groups (control, low, medium and high doses groups). The khat extract was administered orally during (8th -18th) day of gestation. The mothers were sacrificed on the 29th day of gestation. The skeletons of the fetuses were stained with Alizarin Red S and Alcian Blue. Skeletal examination of the fetuses of treated dams showed several types of malformations and variations in all treated groups such as unossified phalanges, deformed sternum, completely unossified vertebral lamina and opened anterior and posterior fontanelles, and reduced length of limb long bones. These findings support the teratogenic effect of the khat on the developing rabbit fetus.
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http://dx.doi.org/10.1080/15513815.2020.1827319DOI Listing
October 2020

A comparative study of two analytical techniques for the simultaneous determination of amprolium HCl and ethopabate from combined dosage form and in presence of their alkaline degradation.

Spectrochim Acta A Mol Biomol Spectrosc 2020 Dec 1;243:118756. Epub 2020 Aug 1.

Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Egypt.

Chemometric and separation-based techniques (HPLC) are the most applicable and versatile analytical techniques for the analysis of multicomponent mixtures, in the present contribution, a comparison was highlighted between the two analytical techniques of utmost importance as stability indicating assays: UV-spectrophotometry and HPLC-UV focusing on the greenness of each for the simultaneous determination of amprolium HCl (AMP) and ethopabate (ETHOP) in the presence of their alkaline degradation products. The first method was chemometric methods applied were PLS-1, GA-PLS and GA-ANN. To compare the prediction ability of the models, a 4-factor 5-level experimental design was used to establish a calibration set of 25 mixtures containing different ratios of the drugs and their degradation products. The validity of the proposed methods was assessed using an independent validation set of 5 mixtures. The comparison between the different models showed the superiority of ANN model in solving the highly overlapped spectra of the quaternary mixture, yet using inexpensive and easy to handle instruments like the UV-VIS spectrophotometer. The ANN method was used for the quantitative analysis of the drugs in pharmaceutical dosage form via handling the UV spectral data. The second method was based on liquid chromatographic HPLC determination of AMP and ETHOP using C18 column (250 × 4.6 mm)-PRONTOSIL 5 μm, a mobile phase consisting of methanol: Hexane sulphonic acid sodium salt at (pH = 3.4 ± 0.2) adjusted by orthophosphoric acid (55: 45 v/v). Quantitation was achieved with UV detection at 270 nm at temperature 24 °C. Linearity, accuracy and precision were found to be acceptable over the concentration range of 10.0-70.0 and 1.0-25.0 μg·mL for AMP and ETHOP, respectively. The proposed methods could be successfully applied for the routine analysis of the studied drugs either in their pure bulk powders or in their pharmaceutical preparations without any preliminary separation step. The results obtained were statistically compared with those obtained by applying the reported method.
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http://dx.doi.org/10.1016/j.saa.2020.118756DOI Listing
December 2020

Point-of-care diagnostics for drugs of abuse in biological fluids: application of a microfabricated disposable copper potentiometric sensor.

Mikrochim Acta 2020 08 7;187(9):491. Epub 2020 Aug 7.

Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr-El-Aini, Cairo, 11562, Egypt.

The major objective of this work was to develop a portable, disposable, cost-effective, and reliable POC solid-state electrochemical sensor based on potentiometric transduction to detect benzodiazepine abuse, mainly diazepam (DZP), in biological fluids. To achieve that, microfabricated Cu electrodes on a printed circuit board modified with the conducting polymer poly(3-octylthiophene) (POT) have been employed as a substrate. This polymer was introduced to enhance the stability of the potential drift (0.9 mV/h) and improve the limit of detection (0.126 nmol mL). Nernstian potentiometric response was achieved for DZP over the concentration range 1.0 × 10 to 5.0 × 10 mol L with a slope of 55.0 ± 0.4 mV/decade and E ~ 478.9 ± 0.9. Intrinsic merits of the proposed sensor include rapid response time (11 ± 2 s) and long life time (3 months). In order to enhance the selectivity of the potentiometric sensor towards the target drug and minimize any false positive results, calix[4]arene (CX4) was impregnated as an ionophore within the PVC plastic ion-sensing membrane. The performance of the POC sensors was assessed using electrochemical methods of analysis and electrochemical impedance spectroscopy as a surface characterization tool. The studied sensors were applied to the potentiometric determination of DZP in different biological fluids (plasma, urine, saliva, and human milk) in the presence of its metabolite with an average recovery of 100.9 ± 1.3%, 99.4 ± 1.0%, 101.8 ± 1.2%, and 99.0 ± 2.0%, respectively. Graphical abstract.
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http://dx.doi.org/10.1007/s00604-020-04445-xDOI Listing
August 2020

Stability-Indicating RP-HPLC and CE Methods for Simultaneous Determination of Bisoprolol and Perindopril in Pharmaceutical Formulation: A Comparative Study.

J Chromatogr Sci 2020 Aug;58(8):747-758

Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo 11562, Egypt.

Two fast, accurate and selective stability-indicating methods were developed and validated for the simultaneous determination of bisoprolol, perindopril and three of their possible degradation products. The first proposed method was a gradient reversed phase-high-performance liquid chromatography (HPLC) method, whereas the second was a capillary electrophoresis method. The structures of the obtained degradation products were elucidated using infrared and mass spectrometry. They were also confirmed to be either a drug impurity in the British Pharmacopoeia or a precursor to such impurity. The linearity for bisoprolol and perindopril was achieved in the range of 1-20 μg mL-1 and 5-30 μg mL-1 for HPLC and capillary electrophoresis methods, respectively. The proposed methods were validated according to the International Conference on Harmonisation guidelines. The HPLC method proved to be more sensitive and succeeded in the quantitative determination of the obtained degradation products. Also, it was able to quantify perindopril impurity up to three times lower than the desired limit set by the British Pharmacopoeia. They were successfully employed in the determination of bisoprolol and perindopril in their combined pharmaceutical formulation.
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http://dx.doi.org/10.1093/chromsci/bmaa031DOI Listing
August 2020

Spectrofluorimetric study on fluorescence quenching of tyrosine and l-tryptophan by the aniracetam cognition enhancer drug: quenching mechanism using Stern-Volmer and double-log plots.

Luminescence 2020 Aug 28;35(5):728-737. Epub 2020 Jan 28.

Pharmaceutical Chemistry Department, National Organization for Drug Control and Research (NODCAR), Giza, Egypt.

A novel approach on fluorescence quenching of tyrosine and l-tryptophan is presented for spectrofluorimetric determination of aniracetam in drug substances and products. The quenching mechanism was investigated using Stern-Volmer plots and ultraviolet spectra figures of quencher-fluorophore mixtures. Binding constant and stoichiometry were calculated using double-log plots. The spectrofluorimetric method was optimized for the experimental conditions affecting fluorescence quenching including fluorophore concentration, diluent, and reaction time. Moreover, the pH-rate profile of aniracetam was studied using simple kinetics and found to be stable within the pH range 5-8. Fluorescence quenching of tyrosine and l-tryptophan were observed on addition of aniracetam in aqueous medium at pH 5.5-6.5. Aniracetam quenched the fluorescence of tyrosine and l-tryptophan in the concentration range 1-20 μg/ml and 0.3-20 μg/ml, respectively, with binomial relationships between quenching values (ΔF) and aniracetam concentration. Limits of detection were found to be 0.10 μg/ml for tyrosine-aniracetam and 0.14 μg/ml for l-tryptophan-aniracetam. Method validation was performed as per ICH guidelines and demonstrated that the developed spectrofluorimetric method was accurate, precise, specific, and suitable for analysis of aniracetam in routine quality control laboratories. All experimental materials and solvents used are eco-friendly, indicating that the cited spectrofluorimetric procedure is an excellent green method.
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http://dx.doi.org/10.1002/bio.3778DOI Listing
August 2020

pharmacological investigation of and using animal models.

Saudi J Biol Sci 2019 Nov 9;26(7):1602-1606. Epub 2018 Oct 9.

Department of Biological Sciences Gomal University Dera Ismail Khan KP, Pakistan.

Experimental based evidence suggests that most of the medicinal plants possess wide-ranging pharmacological and biological activities that may possibly use in treatment of inflammation-related diseases. The current study was aimed to explore the acute toxicity, analgesic, sedative and antipyretic activities of and in mices. In experimental models were used in this study. Acute toxicity was evaluated for 24 h' interval at concentration of 500, 1000, 1500 and 2000 mg/kg. The analgesic activity was estimated by acetic acid induced writhing test. White wood apparatus enclosed in stainless steel was used for sedative experiment and antipyretic activity was evaluated in brewer's yeast induced hyperthermic at 50, 100 and 150 mg/kg i.p. Both plants were found safe at all tested doses. and dose-dependently reduced abdominal constrictions in mice. Both plants exhibited significant ( < 0.0001) sedative effects in dose of 50, 150 and 150 mg/kg. Both plants markedly ( < 0.0001) reduced yeast induced hyperthermia. The inhibitions were dose-dependent and remained significant up to five hours of administration. These investigational results have linked a pharmacological indication for the traditional claim of the drugs to be used as an anti-inflammatory, analgesics and antipyretic agents.
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http://dx.doi.org/10.1016/j.sjbs.2018.10.008DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6864368PMC
November 2019

Strategies for stabilizing formulation and QbD assisted development of robust stability indicating method of azilsartan medoxomil/chlorthalidone.

J Pharm Biomed Anal 2020 Jan 4;178:112910. Epub 2019 Oct 4.

Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, Cairo 11562, Egypt.

Reasons for formulation instability were investigated either encountered during production or analytical processes of azilsartan medoxomil (AZM)/chlorthalidone hydrochloride (CLT) tablets. Through the identification of the most feasible degradation pathways, several strategies were proposed to enhance the stability of AZM/CLT formulation. Furthermore, a robust HPLC-UV method was developed and validated for the determination of AZM, CLT in the presence of their possible degradation products. For chromatographic method development, typical quality by design (QbD) approach was implemented. In order to optimize fourteen chromatographic responses, we have used a central composite design with four factors (pH, temperature, flow rate, and acetonitrile %). However, the developed method provides a design space, but optimum parameters were Inertsil C8 column (150 x 4.6 mm, 5 μm), mobile phase composed of 0.025 M phosphate buffer pH 2.7 and acetonitrile (52.5: 47.5%), with flow rate of 1.5 mL.min and detection wavelength 225 nm at 33 °C. The method was then validated according to ICH guidelines and applied to quantitate AZM and CLT in the pharmaceutical formulation. To the best of our knowledge, this manuscript is the first attempt to discuss such instability issues, to propose strategies that enhance the stability of AZM/CLT tablet formulation, to develop robust stability-indicating method taking into consideration the realistic degradation products in addition to minor ones.
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http://dx.doi.org/10.1016/j.jpba.2019.112910DOI Listing
January 2020

Gross morphological features of the air sacs of the hooded crow (Corvus cornix).

Anat Histol Embryol 2020 Mar 15;49(2):159-166. Epub 2019 Oct 15.

Department of Anatomy and Embryology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt.

Air sacs are considered to be one of the controlling factors of bird behaviour and habits in addition to their roles in ventilation, regulating body temperature, swimming and flight. As a scavenger and an omnivorous flight bird, air sacs of the hooded crow were the focus of this study. Eight healthy, adult hooded crows were used to examine the morphological characteristics of the air sacs, which were examined grossly and with latex and cast preparations. In general, the morphological overview of the hooded crow air sacs is similar to other avian species. We observed nine air sacs; four paired sacs (cervical, cranial thoracic, caudal thoracic and abdominal air sacs) and one unpaired sac; the clavicular air sac. The cervical air sac communicated to the lung through the medioventral bronchus and had three diverticula; intermuscular, subscapular and subcutaneous. The clavicular air sac communicated with lung through the medioventral bronchus and had subscapular, axillary, humeral, subpectoral and sternal diverticula. The cranial and caudal thoracic air sacs were communicated with lung through the lateroventral bronchi and the both sacs did not have any diverticula. The abdominal air sacs were posterior to the caudal thoracic air sacs. The left abdominal sac was the largest air sac. The right and left abdominal sacs gave off branches to diverticula that pneumatized synsacrum. The abdominal air sacs gave off femoral diverticula behind the hip joint as well as perirenal diverticula.
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http://dx.doi.org/10.1111/ahe.12504DOI Listing
March 2020

Schwannoma masquerading as adrenocortical tumor: A case report and review of literature.

Urol Case Rep 2019 Sep 30;26:100926. Epub 2019 May 30.

Johns Hopkins Aramco Healthcare-Urology Department, Saudi Arabia.

Schwannomas arises from retroperitoneal space are rare tumors. Adrenal Schwannomas are often misdiagnosed due to deficient of distinctive radiological findings. We report a case of adrenal schwannoma presented with vague abdominal pain. Initially, the patient was diagnosed as adrenocortical tumor that was treated with robotic adrenalectomy. Histopathological and immuno-histochemical examination revealed schwannoma. We will report the case and review the literature regarding this rare tumor.
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http://dx.doi.org/10.1016/j.eucr.2019.100926DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6597528PMC
September 2019

Protective role of Monotheca buxifolia and Bosea amherstiana against H2O2 -induced DNA damage in human lymphocytes and its effect on oxidative enzymes.

Pak J Pharm Sci 2019 Mar;32(2):601-606

Reproductive Physiology Lab, Department of Animal Sciences, Quaid-i-Azam University, Islamabad, Pakistan.

Experimental based evidence suggests that most of the medicinal plants possess a wide-ranging pharmacological and biological activity that may possibly protect tissues against O-induced damages. The objectives of the current study are: first, to investigate the effects of Monotheca buxifolia and Bosea amherstiana on HO induced DNA damage in human lymphocytes and second, to determine its effect on oxidative enzymes. Cells were treated at concentration of 100μg/mL with both plants. Alkaline Single Cell Gel Electrophoresis/comet assay were used for DNA damage analysis. Activities of antioxidant enzymes TBARS, SOD, CAT and POD were assayed on treatment with the extracts. Both plants species possess the protective role against H2O2-induced lymphocytes DNA. Dichloromethane (DCM) fraction of Monotheca buxifolia (H DNA 94.79±0.29%) and methanolic fraction of Bosea amherstiana (H DNA 93.63±2.23%) possess high protection Significantly decrease occur in status of antioxidant enzymes. This study indicates that both plants have potential in preventing oxidative damages/stress related diseases and would be suitably used as supplements in combination with conventional drug for the treatment of cancer like diseases.
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March 2019

MiR302c, Sp1, and NFATc2 regulate interleukin-21 expression in human CD4+CD45RO+ T lymphocytes.

J Cell Physiol 2019 05 21;234(5):5998-6011. Epub 2018 Oct 21.

Department of Biology, Laboratory of Cancer Biology and Molecular Immunology, Faculty of Sciences-I, Lebanese University, Beirut, Lebanon.

Interleukin-21 (IL-21) is a cytokine with potent regulatory effects on different immune cells. Recently, IL-21 has been contemplated for use in the treatment of cancers. However, the molecular mechanisms regulating human IL-21 gene expression has not yet been described. In this study, we initially studied the promoter region and identified the transcription start site. We thereafter described the essential region upstream of the transcription start site and showed the in vivo binding of NFATc2 and SP1 transcription factors to this region, in addition to their positive role in IL-21 expression. We also studied the role of microRNAs (miRNAs) in regulating IL-21 expression. We, thus, established the miRNA profile of CD4+CD45RO+ versus CD4+CD45RA+ isolated from healthy volunteers and identified a signature composed of 12 differentially expressed miRNAs. We showed that miR-302c is able to negatively regulate IL-21 expression by binding directly to its target site in the 3'-untranslated region. Moreover, after using fresh human CD4-positive T cells, we observed the high acetylation level of histone H4, an observation well in line with the already described high expression of IL-21 in CD4+CD45RO+ versus CD4+CD45RA+ T cells. Altogether, our data identified different molecular mechanisms regulating IL-21 expression.
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http://dx.doi.org/10.1002/jcp.27151DOI Listing
May 2019

Clonidine as a Sole Epidural Adjuvant in Combined Spinal-epidural: Clinical Study.

Anesth Essays Res 2018 Apr-Jun;12(2):309-312

Department of Anaesthesiology, Bangalore Medical College and Research Institute, Bengaluru, Karnataka, India.

Background And Aims: Clonidine has been used as an epidural adjuvant along with local anesthetics; however, its use as a sole epidural adjuvant in combined spinal-epidural (CSE) anesthesia has not been explored; thus, this study aimed to assess the effects of clonidine as a sole epidural adjuvant in CSE on sensory and motor characteristics of 0.5% hyperbaric bupivacaine given by subarachnoid route.

Methodology: A total of 60 patients belonging to the American Society of Anesthesiologists Classes I and II aged 18-60 years were randomized in two groups; group G300 and group GNS. G300 group received 300 mg clonidine and GNS received normal saline through epidural route followed by 15 mg of 0.5% hyperbaric bupivacaine as subarachnoid block. Onset of sensory block (time to T10) and motor block (time to Bromage 3), level of sedation (using Modified Ramsay Sedation Score), and hemodynamic changes were recorded. Two-segment regression, duration of analgesia (time for 1 rescue analgesia), and motor block (time to Bromage 0) were recorded. Student's -test (two-tailed, independent) and Chi-square/Fisher's exact probability test were used for statistical analysis.

Results: The demographic data were comparable between the groups. The onset of sensory and motor block was significantly faster in G300 (sensory-71.63 ± 4.51 s, motor-55.63 ± 2.54 s) as compared to GNS (sensory-90.13 ± 4.88 s, motor-118.43 ± 9.50 s) ( < 0.001 and < 0.001, respectively). The two-segment regression was 199.33 ± 19.11 min and 79 ± 9.77 min in G300 and GNS, respectively, ( < 0.001). Duration of analgesia was 317.90 ± 15.32 min and 207 ± 20.66 min for G300 and GNS, respectively ( < 0.001), and duration of motor block was 409.9 ± 34.87 min and 204 ± 22.79 min for G300 and GNS, respectively ( < 0.001). The side effects such as hypotension and bradycardia were statistically and clinically not significant.

Conclusion: Clonidine used as a sole epidural adjuvant in dose of 300 mg, for infraumbilical surgeries, has significantly faster onset of sensory and motor block with prolonged duration of analgesia and motor blockade and no significant side effects on a conventional subarachnoid block performed with 0.5% hyperbaric bupivacaine.
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http://dx.doi.org/10.4103/aer.AER_18_17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6020595PMC
July 2018

Corrigendum to "Application of wavelet and Fourier transforms as powerful signal processing tools for determination of naltrexone and bupropion in their binary mixture" [Spectrochim. Acta A Mol. Biomol. Spectrosc. 191 (2018) 365-371].

Spectrochim Acta A Mol Biomol Spectrosc 2018 05;196:366

Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo, ET 11562, Egypt; Pharmaceutical Chemistry Department, College of Pharmacy, Prince Sattam Bin-Abdul Aziz University, Al-Kharj 11942, Kingdom of Saudi Arabia.

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http://dx.doi.org/10.1016/j.saa.2018.02.045DOI Listing
May 2018

Application of wavelet and Fuorier transforms as powerful alternatives for derivative spectrophotometry in analysis of binary mixtures: A comparative study.

Spectrochim Acta A Mol Biomol Spectrosc 2018 02 14;191:365-371. Epub 2017 Aug 14.

Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo ET-11562, Egypt; Pharmaceutical Chemistry Department, College of Pharmacy, Prince Sattam Bin-Abdul Aziz University, Al-Kharj, 11942, Saudi Arabia.

Two signal processing methods, namely, Continuous Wavelet Transform (CWT) and the second was Discrete Fourier Transform (DFT) were introduced as alternatives to the classical Derivative Spectrophotometry (DS) in analysis of binary mixtures. To show the advantages of these methods, a comparative study was performed on a binary mixture of Naltrexone (NTX) and Bupropion (BUP). The methods were compared by analyzing laboratory prepared mixtures of the two drugs. By comparing performance of the three methods, it was proved that CWT and DFT methods are more efficient and advantageous in analysis of mixtures with overlapped spectra than DS. The three signal processing methods were adopted for the quantification of NTX and BUP in pure and tablet forms. The adopted methods were validated according to the ICH guideline where accuracy, precision and specificity were found to be within appropriate limits.
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http://dx.doi.org/10.1016/j.saa.2017.08.039DOI Listing
February 2018

Development and validation of LC-MS/MS assay for the simultaneous determination of methotrexate, 6-mercaptopurine and its active metabolite 6-thioguanine in plasma of children with acute lymphoblastic leukemia: Correlation with genetic polymorphism.

J Chromatogr B Analyt Technol Biomed Life Sci 2016 Dec 28;1038:88-94. Epub 2016 Oct 28.

Department of Pediatrics, Hematology-Oncology Division, Faculty of Medicine, Ain-Shams University, Cairo, Egypt.

Individualized therapy is a recent approach aiming to specify dosage regimen for each patient according to its genetic state. Cancer chemotherapy requires continuous monitoring of the plasma concentration levels of active forms of cytotoxic drugs and subsequent dose adjustment. In order to attain optimum therapeutic efficacy, correlation to pharmacogenetics data is crucial. In this study, a specific, accurate and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed for determination of methotrexate (MTX), 6-mercaptopurine (MP) and its metabolite 6-thioguanine nucleotide (TG) in human plasma. Based on the basic character of the studied compounds, solid phase extraction using a strong cation exchanger was found the optimum approach to achieve good extraction recovery. Chromatographic separation was carried out using RP-HPLC and isocratic elution by acetonitrile: 0.1% aqueous formic acid (85:15v/v) with a flow rate of 0.8mL/min at 40°C. The detection was performed by tandem mass spectrometry in MRM mode via electrospray ionization source in positive ionization mode. Analysis was carried out within 1.0min over a concentration range of 6.25-200.00ng/mL for the studied analytes. Validation was carried out according to FDA guidelines for bioanalytical method validation and satisfactory results were obtained. The applicability of the assay for the monitoring of the MTX, MP and TG and subsequent application to personalized therapy was demonstrated in a clinical study on children with acute lymphoblastic leukemia (ALL). Results confirmed the need for implementation of reliable analysis tools for therapeutic dose adjustment.
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http://dx.doi.org/10.1016/j.jchromb.2016.10.035DOI Listing
December 2016

Transmission electron microscopic evaluation of neuronal changes in methylmercury-exposed zebrafish embryos (Danio rerio).

Ultrastruct Pathol 2016 Nov-Dec;40(6):333-341. Epub 2016 Oct 27.

c Department of Veterinary Integrative Biosciences , College of Veterinary Medicine and Biomedical Sciences, Texas A&M University , College Station , TX, USA.

Our work aimed to elucidate the ultrastructural changes associated with brain neurons in wild-type zebrafish embryos exposed to different concentrations of methylmercury. Zebrafish embryos were exposed to one of five concentrations of methylmercury (0 [negative control], 5, 10, 50, and 80 parts per billion) starting at six hours post fertilization (hpf). At 96 hpf, cells in the zebrafish embryo brains were examined using transmission electron microscopy. The developing neurons of the control embryos sowed normal cellular ultrastructure. Few alterations were observed among the neurons of zebrafish embryos exposed to 5 ppb methylmercury. The cells of the embryos exposed to 10 ppb methylmercury showed slight cellular degeneration as demonstrated by the accumulation of electron dens bodies which were presumably lysosomes in different stages of formation. In embryos exposed to 50 ppb methylmercury, the neuronal cytoplasm conained large electron dense lysosomes and the rough endoplasmic reticulum appeared to be reduced and irregular in shape. Furthermore, the embryonic brain neurons exposed to 80 ppb methylmercury showed the most severe ultrastructural changes, including some that were consistent with different stages of the cell death process. Obvious cellular changes were observed in this highest exposure group included: disrupted or degenerating nuclei; fragmentation or vacuolization of mitochondrial cristae; and loss of mitochondrial matrix density. Based on these observations, we conclude that these different morphological patterns of cellular changes may reflect either different stages of the cell death process or different types of cell death due to 24 hours of exposure to 80 ppb methylmercury.
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http://dx.doi.org/10.1080/01913123.2016.1234529DOI Listing
April 2018

Development and validation of HPLC and CE methods for simultaneous determination of amlodipine and atorvastatin in the presence of their acidic degradation products in tablets.

Acta Pharm 2016 Dec;66(4):479-490

Two methods were developed for separation and quantitation of amlodipine (AML) and atorvastatin (ATV) in the presence of their acidic degradation products. The first method was a simple isocratic RP-HPLC method while the second was capillary electrophoresis (CE). Degradation products were obtained by acidic hydrolysis of the two drugs and their structures were elucidated for the first time by IR and MS spectra. Degradation products did not interfere with the determination of either drug and the assays were therefore stability-indicating. The linearity of the proposed methods was established over the ranges 1-50 μg mL-1 for AML and ATV in the HPLC method and in the range of 3-50 and 4-50 μg mL-1 for AML and ATV, respectively, in the CE method. The proposed methods were validated according to ICH guidelines. The methods were successfully applied to estimation of AML and ATV in combined tablets.
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http://dx.doi.org/10.1515/acph-2016-0040DOI Listing
December 2016

Phosphoramidate Ligation of Oligonucleotides in Nanoscale Structures.

Chembiochem 2016 06 25;17(12):1150-5. Epub 2016 May 25.

Institut für Organische Chemie, Universität Stuttgart, 70569, Stuttgart, Germany.

The folding of long DNA strands into designed nanostructures has evolved into an art. Being based on linear chains only, the resulting nanostructures cannot readily be transformed into covalently linked frameworks. Covalently linking strands in the context of folded DNA structures requires a robust method that avoids sterically demanding reagents or enzymes. Here we report chemical ligation of the 3'-amino termini of oligonucleotides and 5'-phosphorylated partner strands in templated reactions that produce phosphoramidate linkages. These reactions produce inter-nucleotide linkages that are isoelectronic and largely isosteric to phosphodiesters. Ligations were performed at three levels of complexity, including the extension of branched DNA hybrids and the ligation of six scaffold strands in a small origami.
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http://dx.doi.org/10.1002/cbic.201600061DOI Listing
June 2016

Incidence of Active HCV infection amongst Blood Donors of Mardan District, Pakistan.

Asian Pac J Cancer Prev 2016 ;17(1):235-8

Department of Biotechnology, Bacha Khan University, Charsadda, KPK-Pakistan E-mail :

Hepatitis C is an ailment of liver caused by hepatitis C virus (HCV) infection. About 3% of the world population is infected by this virus. HCV infection is a leading reason for liver cirrhosis and therefore a major source of hepatocellular carcinoma. The study focused on the incidence of active HCV infection in blood donors of Mardan district of KPK, Pakistan. A total of 5318 blood donors were inspected for the presence of anti-HCV antibodies and HCV-RNA using ICT (immune-chromatographic test), ELISA and RT-PCR at Mardan Medical Complex (MMC), Mardan. Out of these, 157 (2.95%) were positive by ICT, 60 (1.12%) by ELISA and 56 (1.05%) for HCV-RNA. The frequency of active HCV infectivity amongst the blood donors from district Mardan, KPK Pakistan was 1.05 %. Application of strict measures during blood donor selection and use of proper screening assays such as ELISA in place of ICT devices can give a more accurate picture so that the incidence of this viral infection in HCV negative blood recipients can be reduced.
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http://dx.doi.org/10.7314/apjcp.2016.17.1.235DOI Listing
December 2016

Level of Heavy Metals in Two Highly Consumed Fish Species at District Lower Dir, Khyber Pakhtunkhwa, Pakistan.

Pak J Biol Sci 2016 ;19(3):115-121

Division of Pathology, ICAR-Indian Veterinary Research Institute, Izatnagar 243122, Bareilly, Uttar Pradesh, India.

The current study was designed to assess heavy metals' concentration in muscle tissues of two Chinese carps, common carp (Cyprinus carpio) and silver carp (Hypophthalmichthys molitrix), available to consumers in markets at district Lower Dir, Khyber Pakhtunkhwa, Pakistan. Fish specimens were collected from three main markets in the study area namely; Chakdara, Timergara and Khall. Heavy metals including; manganese (Mn), lead (Pb), iron (Fe), copper (Cu), zinc (Zn), cadmium (Cd) and cobalt (Co) were investigated using atomic absorption spectrophotometer. Cobalt was not detected in any of the fish specimens while the rest of the metals were lying within the permissible limits suggested by FAO/WHO and ITS for food/fish consumption. The results showed a statistically significant (p<0.05) difference between both species with respect to the concentration of the accumulated heavy metals. In common carp, the heavy metal accumulation was in order of Fe>Mn>Zn>Pb>Cu>Cd, while in silver carp the order was Fe>Mn>Zn>Pb>Cd>Cu. Higher concentration of Pb, Mn, Zn, Cu and Cd was recorded in muscle of common carp while the concentration of Fe was higher in silver carp, indicating higher potential of accumulation of heavy metals in common carp. Statistically significant (p<0.05) correlation was observed between Pb and Zn in common carp while between Cu and Cd in silver carp. The concentration of heavy metals was in the suggested permissible limits and poses no threat if consumed. In order to maintain the heavy metals level within permissible limits, proper care should be taken along with regular assessment.
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http://dx.doi.org/10.3923/pjbs.2016.115.121DOI Listing
January 2016

Advanced stability indicating chemometric methods for quantitation of amlodipine and atorvastatin in their quinary mixture with acidic degradation products.

Spectrochim Acta A Mol Biomol Spectrosc 2016 Feb 8;154:58-66. Epub 2015 Oct 8.

Department of Analytical Chemistry, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562 Cairo, Egypt.

Two advanced, accurate and precise chemometric methods are developed for the simultaneous determination of amlodipine besylate (AML) and atorvastatin calcium (ATV) in the presence of their acidic degradation products in tablet dosage forms. The first method was Partial Least Squares (PLS-1) and the second was Artificial Neural Networks (ANN). PLS was compared to ANN models with and without variable selection procedure (genetic algorithm (GA)). For proper analysis, a 5-factor 5-level experimental design was established resulting in 25 mixtures containing different ratios of the interfering species. Fifteen mixtures were used as calibration set and the other ten mixtures were used as validation set to validate the prediction ability of the suggested models. The proposed methods were successfully applied to the analysis of pharmaceutical tablets containing AML and ATV. The methods indicated the ability of the mentioned models to solve the highly overlapped spectra of the quinary mixture, yet using inexpensive and easy to handle instruments like the UV-VIS spectrophotometer.
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http://dx.doi.org/10.1016/j.saa.2015.10.007DOI Listing
February 2016

Mean centering of double divisor ratio spectra, a novel spectrophotometric method for analysis of ternary mixtures.

Spectrochim Acta A Mol Biomol Spectrosc 2016 Jan 10;153:132-42. Epub 2015 Aug 10.

Department of Analytical Chemistry, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562 Cairo, Egypt.

A novel spectrophotometric method was developed for determination of ternary mixtures without previous separation, showing significant advantages over conventional methods. The new method is based on mean centering of double divisor ratio spectra. The mathematical explanation of the procedure is illustrated. The method was evaluated by determination of model ternary mixture and by the determination of Amlodipine (AML), Aliskiren (ALI) and Hydrochlorothiazide (HCT) in laboratory prepared mixtures and in a commercial pharmaceutical preparation. For proper presentation of the advantages and applicability of the new method, a comparative study was established between the new mean centering of double divisor ratio spectra (MCDD) and two similar methods used for analysis of ternary mixtures, namely mean centering (MC) and double divisor of ratio spectra-derivative spectrophotometry (DDRS-DS). The method was also compared with a reported one for analysis of the pharmaceutical preparation. The method was validated according to the ICH guidelines and accuracy, precision, repeatability and robustness were found to be within the acceptable limits.
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http://dx.doi.org/10.1016/j.saa.2015.07.109DOI Listing
January 2016

Topical pyruvic acid (70%) versus topical salicylic acid (16.7%) compound in treatment of plantar warts: A randomized controlled trial.

Adv Biomed Res 2015 29;4:113. Epub 2015 May 29.

Department of Dermatology, Skin Diseases and Leishmaniasis Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.

Background: Recurrence rate is considerable with current topical treatments of plantar warts. We compared the efficacy of topical pyruvic acid (70%) with salicylic acid (16.7%) compound in treating multiple plantar warts.

Materials And Methods: In this randomized controlled trial, 60 patients with multiple plantar warts were equally randomized to receive either pyruvic acid 70% or compound salicylic acid solution (salicylic acid 16.7%, lactic acid 16.7%, and collodion 100%) that was topically applied twice a day for 4 weeks. Patients were visited every 2 weeks for 1 month after starting treatment and then every 1 month for another 2 months. The number and size of warts, treatment complications (pain, burning, scar, pigmentation, and crust), and recurrence were evaluated.

Results: Warts' number was decreased by -13.12 ± 25.6% with pyruvic acid and by -23.0 ± 28.0% with compound salicylic acid (P = 0.159) after treatment. Warts' size was decreased by -43.47 ± 57.0% with pyruvic acid and by -37.40 ± 32.76% with compound salicylic acid (P = 0.615) after treatment. There was no difference between the two groups in cumulative incidence of treatment complications (P > 0.05). Also, there was no difference between the two groups in recurrence rate at 2 months (10 vs. 16.7%, P = 0.500) or at 3 months after treatment (3.3 vs. 6.7%, P = 0.335).

Conclusion: Topical pyruvic acid and compound salicylic acid had the same efficacy and complications in treating plantar warts. Decision for choosing the treatment can be made based on the costs and individual factors as well as patients preferences.
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http://dx.doi.org/10.4103/2277-9175.157833DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4513322PMC
August 2015

Continuous Wavelet Transform, a powerful alternative to Derivative Spectrophotometry in analysis of binary and ternary mixtures: A comparative study.

Spectrochim Acta A Mol Biomol Spectrosc 2015 Dec 30;151:945-55. Epub 2015 Jun 30.

Department of Analytical Chemistry, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562 Cairo, Egypt.

A comparative study was established between two signal processing techniques showing the theoretical algorithm for each method and making a comparison between them to indicate the advantages and limitations. The methods under study are Numerical Differentiation (ND) and Continuous Wavelet Transform (CWT). These methods were studied as spectrophotometric resolution tools for simultaneous analysis of binary and ternary mixtures. To present the comparison, the two methods were applied for the resolution of Bisoprolol (BIS) and Hydrochlorothiazide (HCT) in their binary mixture and for the analysis of Amlodipine (AML), Aliskiren (ALI) and Hydrochlorothiazide (HCT) as an example for ternary mixtures. By comparing the results in laboratory prepared mixtures, it was proven that CWT technique is more efficient and advantageous in analysis of mixtures with severe overlapped spectra than ND. The CWT was applied for quantitative determination of the drugs in their pharmaceutical formulations and validated according to the ICH guidelines where accuracy, precision, repeatability and robustness were found to be within the acceptable limit.
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http://dx.doi.org/10.1016/j.saa.2015.06.100DOI Listing
December 2015

Different signal processing techniques of ratio spectra for spectrophotometric resolution of binary mixture of bisoprolol and hydrochlorothiazide; a comparative study.

Spectrochim Acta A Mol Biomol Spectrosc 2015 Apr 3;140:334-43. Epub 2015 Jan 3.

Department of Analytical Chemistry, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562 Cairo, Egypt.

Five signal processing techniques were applied to ratio spectra for quantitative determination of bisoprolol (BIS) and hydrochlorothiazide (HCT) in their binary mixture. The proposed techniques are Numerical Differentiation of Ratio Spectra (ND-RS), Savitsky-Golay of Ratio Spectra (SG-RS), Continuous Wavelet Transform of Ratio Spectra (CWT-RS), Mean Centering of Ratio Spectra (MC-RS) and Discrete Fourier Transform of Ratio Spectra (DFT-RS). The linearity of the proposed methods was investigated in the range of 2-40 and 1-22 μg/mL for BIS and HCT, respectively. The proposed methods were applied successfully for the determination of the drugs in laboratory prepared mixtures and in commercial pharmaceutical preparations and standard deviation was less than 1.5. The five signal processing techniques were compared to each other and validated according to the ICH guidelines and accuracy, precision, repeatability and robustness were found to be within the acceptable limit.
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http://dx.doi.org/10.1016/j.saa.2014.12.107DOI Listing
April 2015

Different approaches in Partial Least Squares and Artificial Neural Network models applied for the analysis of a ternary mixture of Amlodipine, Valsartan and Hydrochlorothiazide.

Spectrochim Acta A Mol Biomol Spectrosc 2014 Mar 20;122:744-50. Epub 2013 Nov 20.

Department of Analytical Chemistry, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562 Cairo, Egypt.

Different chemometric models were applied for the quantitative analysis of Amlodipine (AML), Valsartan (VAL) and Hydrochlorothiazide (HCT) in ternary mixture, namely, Partial Least Squares (PLS) as traditional chemometric model and Artificial Neural Networks (ANN) as advanced model. PLS and ANN were applied with and without variable selection procedure (Genetic Algorithm GA) and data compression procedure (Principal Component Analysis PCA). The chemometric methods applied are PLS-1, GA-PLS, ANN, GA-ANN and PCA-ANN. The methods were used for the quantitative analysis of the drugs in raw materials and pharmaceutical dosage form via handling the UV spectral data. A 3-factor 5-level experimental design was established resulting in 25 mixtures containing different ratios of the drugs. Fifteen mixtures were used as a calibration set and the other ten mixtures were used as validation set to validate the prediction ability of the suggested methods. The validity of the proposed methods was assessed using the standard addition technique.
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http://dx.doi.org/10.1016/j.saa.2013.11.045DOI Listing
March 2014
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