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Chemical composition, antioxidant and cytotoxicity activities of the essential oils of Myristica fragrans and Morinda citrifolia

Piaru SP, Mahmud R, Abdul Majid AM, Ismail S, Man CN

J Sci Food Agric, 2012 Feb;92(3):593-7.
PMID: 25520982

In this study the chemical composition, antioxidant activities and cytotoxic effect of the essential oils of Myristica fragrans (nutmeg) and Morinda citrifolia (mengkudu) were determined.
Fulltext Seroprevalence and sources of toxoplasmosis among Orang Asli (indigenous) communities in Peninsular Malaysia

Ngui R, Lim YA, Amir NF, Nissapatorn V, Mahmud R

Am J Trop Med Hyg, 2011 Oct;85(4):660-6.
PMID: 21976569 DOI: 10.4269/ajtmh.2011.11-0058

This study aims to evaluate the current seroprevalence of toxoplasmosis among indigenous communities in Peninsular Malaysia and relate its association with epidemiological data. Overall seroprevalence of Toxoplasma gondii was 37.0% with 31.0% immunoglobulin (Ig) G, 1.8% IgM, and 4.2% seropositivity for both anti-Toxoplasma antibodies. Multivariate analysis showed that age above 12 years (odds ratio [OR] = 2.70, 95% confidence interval [CI] = 1.75-4.04, P < 0.001), using untreated river and mountain water supplies (OR = 1.50, 95% CI = 1.01-2.40, P = 0.050), and close proximity with cats (OR = 1.40, 95% CI = 1.10-1.76, P = 0.010) were factors associated with toxoplasmosis. Given the high seroprevalence of toxoplasmosis among these communities who live in poor socioeconomic conditions, a comprehensive health surveillance program and screening should be initiated among women of childbearing age and pregnant women during the antenatal period for early diagnosis and treatment. The role of domestic cats and environmental contamination with oocyst in soil and water has to be highlighted and addressed in future prevention strategies for these communities.
Evaluation of the use of Cocos nucifera as antimalarial remedy in Malaysian folk medicine

Al-Adhroey AH, Nor ZM, Al-Mekhlafi HM, Amran AA, Mahmud R

J Ethnopharmacol, 2011 Apr 12;134(3):988-91.
PMID: 21277969 DOI: 10.1016/j.jep.2011.01.026

White flesh extract of Cocos nucifera (coconut) was studied to ascertain the ethnopharmacological standing of its antimalarial usage in Malaysian folk medicine.
Improvement of digital mammogram images using histogram equalization, histogram stretching and median filter

Langarizadeh M, Mahmud R, Ramli AR, Napis S, Beikzadeh MR, Rahman WE

J Med Eng Technol, 2011 Feb;35(2):103-8.
PMID: 21204610 DOI: 10.3109/03091902.2010.542271

Breast cancer is one of the most important diseases in females worldwide. According to the Malaysian Oncological Society, about 4% of women who are 40 years old and above are involved have breast cancer. Masses and microcalcifications are two important signs of breast cancer diagnosis on mammography. Enhancement techniques, i.e. histogram equalization, histogram stretching and median filters, were used to provide better visualization for radiologists in order to help early detection of breast abnormalities. In this research 60 digital mammogram images which includes 20 normal and 40 confirmed diagnosed cancerous cases were selected and manipulated using the mentioned techniques. The original and manipulated images were scored by three expert radiologists. Results showed that the selected methods have a positive significant effect on image quality.
Fulltext Plasmodium ovale infection in Malaysia: first imported case

Lim YA, Mahmud R, Chew CH, T T, Chua KH

Malar J, 2010;9:272.
PMID: 20929588 DOI: 10.1186/1475-2875-9-272

BACKGROUND:
Plasmodium ovale infection is rarely reported in Malaysia. This is the first imported case of P. ovale infection in Malaysia which was initially misdiagnosed as Plasmodium vivax.

METHODS:
Peripheral blood sample was first examined by Giemsa-stained microscopy examination and further confirmed using a patented in-house multiplex PCR followed by sequencing.

RESULTS AND DISCUSSION:
Initial results from peripheral blood smear examination diagnosed P. vivax infection. However further analysis using a patented in-house multiplex PCR followed by sequencing confirmed the presence of P. ovale. Given that Anopheles maculatus and Anopheles dirus, vectors of P. ovale are found in Malaysia, this finding has significant implication on Malaysia's public health sector.

CONCLUSIONS:
The current finding should serve as an alert to epidemiologists, clinicians and laboratory technicians in the possibility of finding P. ovale in Malaysia. P. ovale should be considered in the differential diagnosis of imported malaria cases in Malaysia due to the exponential increase in the number of visitors from P. ovale endemic regions and the long latent period of P. ovale. It is also timely that conventional diagnosis of malaria via microscopy should be coupled with more advanced molecular tools for effective diagnosis.
Zoonotic Brugia pahangi filariasis in a suburbia of Kuala Lumpur City, Malaysia

Tan LH, Fong MY, Mahmud R, Muslim A, Lau YL, Kamarulzaman A

Parasitol Int, 2011 Jan;60(1):111-3.
PMID: 20951228 DOI: 10.1016/j.parint.2010.09.010

Five local Malaysian patients with clinical manifestations consistent with lymphatic filariasis were referred to our medical centre between 2003 and 2006. Although no microfilariae (mf) were detected in their nocturnal blood samples, all were diagnosed to have lymphatic filariasis on the basis of clinical findings and positive serology results. PCR on their blood samples revealed that two of the patients were infected with Brugia pahangi, an animal filarial worm hitherto not known to cause human disease in the natural environment. All the patients were successfully treated with anti-filarial drugs: four patients were treated with a combination of diethylcarbamazine (DEC) and albendazole, and one with doxycycline. Four of them were residents of Petaling Jaya, a residential suburbia located 10 km southwest of Kuala Lumpur city, Malaysia. The fifth patient was a frequent visitor of the suburbia. This suburbia has no history or record of B. malayi infection. The most likely vector of the worm was Armigeres subalbatus as extensive entomological surveys within the suburbia revealed only adult females of this mosquito species were infected with B. pahangi larvae. Wild monkeys caught in the suburbia were free from B. pahangi mf, but domestic cats were mf positive. This suggests that infected cats might be the source of the zoonotic infection in the suburbia.
Multicentre evaluations of two new rapid IgG4 tests (WB rapid and panLF rapid) for detection of lymphatic filariasis

Noordin R, Itoh M, Kimura E, Abdul Rahman R, Ravindran B, Mahmud R, et al.

Filaria journal, 2007;6:9.
PMID: 17961262

In the global effort to eliminate lymphatic filariasis (LF), rapid field-applicable tests are useful tools that will allow on-site testing to be performed in remote places and the results to be obtained rapidly. Exclusive reliance on the few existing tests may jeopardize the progress of the LF elimination program, thus the introduction of other rapid tests would be useful to address this issue. Two new rapid immunochromatographic IgG4 cassette tests have been produced, namely WB rapid and panLF rapid, for detection of bancroftian filariasis and all three species of lymphatic filaria respectively. WB rapid was developed using BmSXP recombinant antigen, while PanLF rapid was developed using BmR1 and BmSXP recombinant antigens. A total of 165 WB rapid and 276 panLF rapid tests respectively were evaluated at USM and the rest were couriered to another university in Malaysia (98 WB rapid, 129 panLF rapid) and to universities in Indonesia (56 WB rapid, 62 panLF rapid), Japan (152 of each test) and India (18 of each test) where each of the tests underwent independent evaluations in a blinded manner. The average sensitivities of WB rapid and panLF rapid were found to be 97.6% (94%-100%) and 96.5% (94%-100%) respectively; while their average specificities were both 99.6% (99%-100%). Thus this study demonstrated that both the IgG4 rapid tests were highly sensitive and specific, and would be useful additional tests to facilitate the global drive to eliminate this disease.
Management of iatrogenic facial nerve palsy and labyrinthine fistula in mastoid surgery

Long YT, bin Sabir Husin Athar PP, Mahmud R, Saim L

Asian J Surg, 2004 Jul;27(3):176-9.
PMID: 15564156

A 6-year review of complications of mastoid surgery between June 1995 and June 2001 revealed five cases with serious iatrogenic complications from mastoid surgery, of which four were facial nerve palsy and two were labyrinthine fistula. One of these patients had concomitant facial nerve palsy and labyrinthine fistula. There were two cases of complete facial nerve palsy (House Brackmann grade VI) and two cases of incomplete palsy (House Brackmann grades IV and V). The second genu was the site of injury in three of the four cases. Of the four cases with facial nerve palsy, two patients had full recovery (House Brackmann grade I), one recovered only to House Brackmann grade III, and one was lost to follow-up. Both patients with labyrinthine fistula had postoperative vertigo and profound sensorineural hearing loss. The site of iatrogenic fenestration was the lateral semicircular canal in both cases.
Fulltext In vitro and in vivo effects of standardized extract and fractions of Phaleria macrocarpa fruits pericarp on lead carbohydrate digesting enzymes

Ali RB, Atangwho IJ, Kuar N, Ahmad M, Mahmud R, Asmawi MZ

BMC Complement Altern Med, 2013;13:39.
PMID: 23425283 DOI: 10.1186/1472-6882-13-39

One vital therapeutic approach for the treatment of type 2 diabetes mellitus is the use of agents that can decrease postprandial hyperglycaemia by inhibiting carbohydrate digesting enzymes. The present study investigated the effects of bioassay-guided extract and fractions of the dried fruit pericarp of Phaleria macrocarpa, a traditional anti-diabetic plant, on α-glucosidase and α-amylase, in a bid to understand their anti-diabetic mechanism, as well as their possible attenuation action on postprandial glucose increase.
Fulltext Nutrigenomic effects of edible bird's nest on insulin signaling in ovariectomized rats

Hou Z, Imam MU, Ismail M, Ooi DJ, Ideris A, Mahmud R

Drug Des Devel Ther, 2015;9:4115-25.
PMID: 26316695 DOI: 10.2147/DDDT.S80743

Estrogen deficiency alters quality of life during menopause. Hormone replacement therapy has been used to improve quality of life and prevent complications, but side effects limit its use. In this study, we evaluated the use of edible bird's nest (EBN) for prevention of cardiometabolic problems in rats with ovariectomy-induced menopause. Ovariectomized female rats were fed for 12 weeks with normal rat chow, EBN, or estrogen and compared with normal non-ovariectomized rats. Metabolic indices (insulin, estrogen, superoxide dismutase, malondialdehyde, oral glucose tolerance test, and lipid profile) were measured at the end of the experiment from serum and liver tissue homogenate, and transcriptional levels of hepatic insulin signaling genes were measured. The results showed that ovariectomy worsened metabolic indices and disrupted the normal transcriptional pattern of hepatic insulin signaling genes. EBN improved the metabolic indices and also produced transcriptional changes in hepatic insulin signaling genes that tended toward enhanced insulin sensitivity, and glucose and lipid homeostasis, even better than estrogen. The data suggest that EBN could meliorate estrogen deficiency-associated increase in risk of cardiometabolic disease in rats, and may in fact be useful as a functional food for the prevention of such a problem in humans. The clinical validity of these findings is worth studying further.
Fulltext Fatal case of amoebic liver abscess in a child

Ng KF, Tan KK, Ngui R, Lim YA, Amir A, Rajoo Y, et al.

Asian Pac J Trop Med, 2015 Oct;8(10):878-80.
PMID: 26522307 DOI: 10.1016/j.apjtm.2015.09.018

We reported a case of amoebic liver abscess (ALA) in a 6-year-old Malaysian boy who presented with fever, lethargy, diarrhoea and right hypochondriac pain. On admission he was diagnosed with perforated acute appendicitis and a laparotomy was done. After surgery he developed acute respiratory distress. Ultrasonography, chest X-Ray and CT scan revealed two ALAs in the posterior segment of right lobe of liver, pleural effusion and collapsed consolidation of lungs bilaterally. Percutaneous liver abscesses drainage was done and intravenous Metronidazole was started. PCR carried out on the pus from the abscess was positive for Entamoeba histolytica. Patient however succumbed to the infection one week after admission.
Fulltext First Case Report of Canthariasis in an Infant Caused by the Larvae of Lasioderma serricorne (Coleoptera: Anobiidae)

Mokhtar AS, Sridhar GS, Mahmud R, Jeffery J, Lau YL, Wilson JJ, et al.

J Med Entomol, 2016 Sep 01;53(5):1234-1237.
PMID: 27208008 DOI: 10.1093/jme/tjw071

We report an unusual cause of gastrointestinal infection occurring in a 1-year-old infant patient who was brought to a public hospital in Kuala Lumpur, Malaysia. Larvae passed out in the patient's feces were confirmed by DNA barcoding as belonging to the species, Lasioderma serricorne (F.), known as the cigarette beetle. We postulate that the larvae were acquired from contaminated food and were responsible for gastrointestinal symptoms in the patient. To our knowledge, this the first report of human canthariasis caused by larvae of L. serricorne.
Effects of edible bird's nest on hippocampal and cortical neurodegeneration in ovariectomized rats

Zhiping H, Imam MU, Ismail M, Ismail N, Yida Z, Ideris A, et al.

Food Funct, 2015 May;6(5):1701-11.
PMID: 25920003 DOI: 10.1039/c5fo00226e

The aim of this research is to investigate whether edible bird's nest (EBN) attenuates cortical and hippocampal neurodegeneration in ovariectomized rats. Ovariectomized rats were randomly divided into seven experimental groups (n = 6): the ovariectomy (OVX) group had their ovaries surgically removed; the sham group underwent surgical procedure similar to OVX group, but ovaries were left intact; estrogen group had OVX and received estrogen therapy (0.2 mg kg(-1) per day); EBN treatment groups received 6%, 3%, and 1.5% EBN, respectively. Control group was not ovariectomized. After 12 weeks of intervention, biochemical assays were performed for markers of neurodegeneration, and messenger ribonucleic acid (mRNA) levels of oxidative stress-related genes in the hippocampus and frontal cortex of the brain were analysed. Caspase 3 (cysteine-aspartic proteases 3) protein levels in the hippocampus and frontal cortex were also determined using western blotting. The results show that EBNs significantly decreased estrogen deficiency-associated serum elevation of advanced glycation end-products (AGEs), and they changed redox status as evidenced by oxidative damage (malondialdehyde content) and enzymatic antioxidant defense (superoxide dismutase and catalase) markers. Furthermore, genes associated with neurodegeneration and apoptosis were downregulated in the hippocampus and frontal cortex by EBN supplementation. Taken together, the results suggest that EBN has potential for neuroprotection against estrogen deficiency-associated senescence, at least in part via modification of the redox system and attenuation of AGEs.
Detection of human malaria using recombinant Plasmodium knowlesi merozoire surface protein-1 (MSP-1₁₉) expressed in Escherichia coli

Sonaimuthu P, Cheong FW, Chin LC, Mahmud R, Fong MY, Lau YL

Exp Parasitol, 2015 Jun;153:118-22.
PMID: 25812552 DOI: 10.1016/j.exppara.2015.03.010

Malaria remains one of the world's most important infectious diseases and is responsible for enormous mortality and morbidity. Human infection with Plasmodium knowlesi is widely distributed in Southeast Asia. Merozoite surface protein-1₁₉ (MSP-1₁₉), which plays an important role in protective immunity against asexual blood stage malaria parasites, appears as a leading immunogenic antigen of Plasmodium sp. We evaluated the sensitivity and specificity of recombinant P. knowlesi MSP-1₁₉ (rMSP-1₁₉) for detection of malarial infection. rMSP-1₁₉ was expressed in Escherichia coli expression system and the purified rMSP-1₁₉ was evaluated with malaria, non-malaria and healthy human serum samples (n = 215) in immunoblots. The sensitivity of rMSP-1₁₉ for detection of P. knowlesi, Plasmodium falciparum, Plasmodium vivax and Plasmodium ovale infection was 95.5%, 75.0%, 85.7% and 100%, respectively. rMSP-1₁₉ did not react with all the non-malaria and healthy donor sera, which represents 100% specificity. The rMSP-1₁₉ could be used as a potential antigen in serodiagnosis of malarial infection in humans.
Toxoplasmosis in HIV/AIDS patients: a current situation

Nissapatorn V, Lee C, Quek KF, Leong CL, Mahmud R, Abdullah KA

Jpn J Infect Dis, 2004 Aug;57(4):160-5.
PMID: 15329448

The seroprevalence of toxoplasmosis among 505 of human immunodeficiency virus (HIV)/AIDS patients was 226 (44.8%; 95% CI 42.64-51.76): 27 (47.4%) and 199 (44.4%) showed Toxoplasma seropositivity with and without toxoplasmic encephalitis (TE), respectively (P <0.05). The majority of these patients were in the 25-34 age group (44 versus 39%), male (86 versus 76%), and Chinese (49 versus 53%), though no statistical significance was found between the two. Significant differences between these two groups were noted, however, in terms of marital status, occupation, and present address. The heterosexual exhibited the most frequent behavior at risk for HIV infection, and accounted for 51 and 59% of patients with and without TE, respectively. Only 17/260 (6.5%) and 1/137 (0.7%) of them later acquired TE after receiving primary chemoprophylaxis (cotrimoxazole) and antiretroviral therapy including HAART (P <0.05). Fifty-seven (11.3%) out of those 505 patients were diagnosed with AIDS-related TE. The most common clinical manifestation was headache (56%). The computed tomography scan findings showed most lesions to be multiple (96.4%), hypodense (66.7%), and in the parietal region (39.3%). Twenty-seven (47.4%) patients had chronic (latent) Toxoplasma infection as evidenced by seropositivity for anti-Toxoplasma (IgG) antibody. At the time of diagnosis, the range of CD4 cell count was from 0-239 with a median of 25 cells/cumm. We also found that a CD4 count of less than 100 cells/cumm was significantly associated with development of TE (P <0.05). Clinical outcomes showed that among those who survived, 21 (36.8%), 16 (28.1%), and 2 (3.5%) of patients had completed treatment, transferred out, and were lost to follow up, respectively. Unfortunately, 18 (31.6%) of the cases were officially pronounced dead. Overall, 7 (12.3%) patients were detected as recurrent TE in this study.
Lactoferrin and ovotransferrin contribute toward antioxidative effects of Edible Bird's Nest against hydrogen peroxide-induced oxidative stress in human SH-SY5Y cells

Hou Z, Imam MU, Ismail M, Azmi NH, Ismail N, Ideris A, et al.

Biosci Biotechnol Biochem, 2015;79(10):1570-8.
PMID: 26057702 DOI: 10.1080/09168451.2015.1050989

There are reports of improved redox outcomes due to consumption of Edible Bird's Nest (EBN). Many of the functional effects of EBN can be linked to its high amounts of antioxidants. Interestingly, dietary components with high antioxidants have shown promise in the prevention of aging and its related diseases like Alzheimer's disease. In this study, the antioxidative potentials of EBN and its constituents, lactoferrin (LF) and ovotransferrin (OVF), were determined and protective effects against hydrogen peroxide (H2O2)- induced toxicity on SH-SY5Y cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and acridine orange and propidium iodide (AO/PI) staining with microscopy were examined. Results showed that EBN and its constituents attenuated H2O2-induced cytotoxicity, and decreased radical oxygen species (ROS) through increased scavenging activity. Furthermore, LF, OVF, and EBN produced transcriptional changes in antioxidant related genes that tended towards neuroprotection as compared to H2O2-treated group. Overall, the results suggest that LF and OVF may produce synergistic or all-or-none antioxidative effects in EBN.
Fulltext Evaluation of codon optimized recombinant Plasmodium knowlesi merozoite surface protein-119 (pkMSP-119) expressed in Pichia pastoris

Lau YL, Cheong FW, Chin LC, Mahmud R, Chen Y, Fong MY

Trop Biomed, 2014 Dec;31(4):749-59.
PMID: 25776601 MyJurnal

Malaria causes high global mortality and morbidity annually. Plasmodium knowlesi has been recognised as the fifth human Plasmodium sp. and its infection is widely distributed in Southeast Asia. Merozoite surface protein-119 (MSP-119) appears as a potential candidate for malaria blood stage vaccine as it could induce protective immunity. In this study, codon optimized P. knowlesi MSP-119 (pkMSP-119) was expressed and purified in yeast Pichia pastoris expression system. The purified recombinant protein was further evaluated using Western blot assay using knowlesi malaria, non-knowlesi human malaria, non-malarial parasitic infections and healthy serum samples (n = 50). The sensitivity of purified pkMSP-119 towards detection of knowlesi infection was as 28.6% (2/7). pkMSP-119 did not react with all non-malarial parasitic infections and healthy donor sera, yet reacted with some non-knowlesi human malaria sera, therefore lead to a specificity of 86.0% (37/43).
Fulltext Acanthamoeba genotype T4 detected in naturally-infected feline corneas found to be in homology with those causing human keratitis

Ithoi I, Mahmud R, Abdul Basher MH, Jali A, Abdulsalam AM, Ibrahim J, et al.

Trop Biomed, 2013 Mar;30(1):131-40.
PMID: 23665719 MyJurnal

A total of 10 out of 65 cornea swab samples from cats with eye symptoms showed Acanthamoeba-like morphology after cultivation. By PCR and DNA sequencing of Acanthamoeba diagnostic fragment 3 (DF3), all 10 isolates from the positive samples were categorized into two homologous groups of AfC1 (PM1, PM2, PM3, PF6, KM7, KF8, KMK9) and AfC2 (PM4, PM5, KFK10) due to the presence of bases A(354) and G(354), respectively. Furthermore, DF3 of AfC1 and AfC2 showed 100% similarity with Genbank reference isolates with the accession numbers DQ087314, EU146073 and U07401, GU808323, which were Acanthamoeba castellanii strains genotype T4 originating from human keratitis. This finding suggests that A. castellani strains have the capability to infect cats and human under favorable conditions.
Estimation of body height from spinal length measurements using post-mortem computed tomographic images

Zyoud TYT, Abdul Rashid SN, Suppiah S, Mahmud R, Kabeer A, Abd Manaf R, et al.

Malays J Pathol, 2020 Dec;42(3):423-431.
PMID: 33361724

INTRODUCTION: Post-mortem computed tomography (PMCT) provides information that helps in the determination of the cause of death and corpse identification of disaster victims. One of the methods for corpse identification includes assessment of the body stature. There is a lack of post-mortem imaging studies that focus on the anthropometric assessment of corpses. Our aim was to identify the relationship between cadaveric spine length and autopsy length (AL) among and autopsy length (AL) among a Malaysian population and derive a regression formula for the estimation of corpse body height using PMCT.
MATERIALS AND METHODS: We retrospectively assessed 107 cadavers that had undergone conventional autopsy and PMCT. We made 5 measurements from the PMCT that included cervical length (CL), thoracic length (TL), lumbosacral length (LS), total column length of the spine, excluding the sacrum and coccyx (TCL), and ellipse line measurement of the whole spine, excluding the sacrum and coccyx (EL). We compared these anthropometric PMCT measurements with AL and correlated them using linear regression analysis.
RESULTS: The results showed a significant linear relationship existed between TL and LS with AL, which was higher in comparison with the other parameters than the rest of the spine parameters. The linear regression formula derived was: 48.163 + 2.458 (TL) + 2.246 (LS).
CONCLUSIONS: The linear regression formula derived from PMCT spine length parameters particularly thoracic and lumbar spine gave a finer correlation with autopsy body length and can be used for accurate estimation of cadaveric height. To the best of our knowledge, this is the first ever linear regression formula for cadaveric height assessment using only post mortem CT spine length measurements.
Molecular markers associated with resistance to commonly used antimalarial drugs among Plasmodium falciparum isolates from a malaria-endemic area in Taiz governorate-Yemen during the transmission season

Alareqi LMQ, Mahdy MAK, Lau YL, Fong MY, Abdul-Ghani R, Mahmud R

Acta Trop, 2016 Oct;162:174-179.
PMID: 27343362 DOI: 10.1016/j.actatropica.2016.06.016

Since 2005, artesunate (AS) plus sulfadoxine/pyrimethamine (SP) combination has been adopted as the first-line treatment for uncomplicated malaria in Yemen in response to the high level of Plasmodium falciparum resistance to chloroquine (CQ). Therefore, the aim of the present study was to determine the frequency distribution of molecular markers associated with resistance to CQ and AS plus SP combination among P. falciparum isolates from a malaria-endemic area in Taiz governorate, Yemen. Fifty P. falciparum isolates were collected during a cross-sectional study in Mawza district, Taiz, in the period from October 2013 to April 2014. The isolates were investigated for drug resistance-associated molecular markers in five genes, including P. falciparum CQ resistance transporter (pfcrt) 76T and P. falciparum multidrug resistance 1 (pfmdr1) 86Y as markers of resistance to CQ, mutations in the Kelch 13 (K13) propeller domain for resistance to AS, and P. falciparum dihydrofolate reductase (pfdhfr) and P. falciparum dihydropteroate synthase (pfdhps) genes for resistance to SP. Nested polymerase chain reaction was used to amplify target genes in DNA extracts of the isolates followed by restriction fragment length polymorphism for detecting 76T and 86Y mutations in pfcrt and pfmdr1, respectively, and by DNA sequencing for detecting mutations in K13, pfdhfr and pfdhps. All the investigated isolates from Mawza district were harboring the pfcrt 76T mutant and the pfmdr1 N86 wild-type alleles. The pfdhfr 51I/108N double mutant allele was found in 2.2% (1/45) of the isolates; however, no mutations were detected at codons 436, 437, 540, 581 and 613 of pfdhps. All P. falciparum isolates that were successfully sequenced (n=47) showed the K13 Y493, R539, I543 and C580 wild-type alleles. In conclusion, the pfcrt 76T mutant allele is fixed in the study area about six years after the official withdrawal of CQ, possibly indicating its over-the-counter availability and continued use as a self-medication in the study area. However, the almost predominant wild-type alleles of the genes associated with resistance to AS and SP among P. falciparum isolates in the present study indicates the sustained efficacy of the currently adopted first-line treatment of AS plus SP in the study area.

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