Swedish traveller with Plasmodium knowlesi malaria after visiting Malaysian Borneo

Plasmodium knowlesi is typically found in nature in macaques and has recently been recognized as the fifth species of Plasmodium causing malaria in human populations in south-east Asia. A case of knowlesi malaria is described in a Swedish man, who became ill after returning from a short visit to Malaysian Borneo in October 2006. His P. knowlesi infection was not detected using a rapid diagnostic test for malaria, but was confirmed by PCR and molecular characterization. He responded rapidly to treatment with mefloquine. Evaluation of rapid diagnostic kits with further samples from knowlesi malaria patients are necessary, since early identification and appropriate anti-malarial treatment of suspected cases are essential due to the rapid growth and potentially life-threatening nature of P. knowlesi. Physicians should be aware that knowlesi infection is an important differential diagnosis in febrile travellers, with a recent travel history to forested areas in south-east Asia, including short-term travellers who tested negative with rapid diagnostic tests

A TaqMan real-time PCR assay for the detection and quantitation of Plasmodium knowlesi

<p>Abstract</p> <p>Background</p> <p>The misdiagnosis of <it>Plasmodium knowlesi </it>by microscopy has prompted a re-evaluation of the geographic distribution, prevalence and pathogenesis of this species using molecular diagnostic tools. In this report, a specific probe for <it>P. knowlesi</it>, that can be used in a previously described TaqMan real-time PCR assay for detection of <it>Plasmodium </it>spp., and <it>Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae </it>and <it>Plasmodium ovale</it>, was designed and validated against clinical samples.</p> <p>Methods</p> <p>A hydrolysis probe for a real-time PCR assay was designed to recognize a specific DNA sequence within the <it>P. knowlesi </it>small subunit ribosomal RNA gene. The sensitivity, linearity and specificity of the assay were determined using plasmids containing <it>P. knowlesi </it>DNA and genomic DNA of <it>P. falciparum, P. knowlesi, P. malariae, P. ovale </it>and <it>P. vivax </it>isolated from clinical samples. DNA samples of the simian malaria parasites <it>Plasmodium cynomolgi </it>and <it>Plasmodium inui </it>that can infect humans under experimental conditions were also examined together with human DNA samples.</p> <p>Results</p> <p>Analytical sensitivity of the <it>P. knowlesi</it>-specific assay was 10 copies/μL and quantitation was linear over a range of 10-10⁶copies. The sensitivity of the assay is equivalent to nested PCR and <it>P. knowlesi </it>DNA was detected from all 40 clinical <it>P. knowlesi </it>specimens, including one from a patient with a parasitaemia of three parasites/μL of blood. No cross-reactivity was observed with 67 <it>Plasmodium </it>DNA samples (31 <it>P. falciparum</it>, 23 <it>P. vivax</it>, six <it>P. ovale</it>, three <it>P. malariae</it>, one <it>P. malariae/P. ovale</it>, one <it>P. falciparum/P. malariae, one P. inui and one P. cynomolgi) </it>and four samples of human DNA.</p> <p>Conclusions</p> <p>This test demonstrated excellent sensitivity and specificity, and adds <it>P. knowlesi </it>to the repertoire of <it>Plasmodium </it>targets for the clinical diagnosis of malaria by real-time PCR assays. Furthermore, quantitation of DNA copy number provides a useful advantage over other molecular assays to investigate the correlation between levels of infection and the spectrum of disease.</p

Clinical and parasitological response to oral chloroquine and primaquine in uncomplicated human Plasmodium knowlesi infections

BACKGROUND: Plasmodium knowlesi is a cause of symptomatic and potentially fatal infections in humans. There are no studies assessing the detailed parasitological response to treatment of knowlesi malaria infections in man and whether antimalarial resistance occurs. METHODS: A prospective observational study of oral chloroquine and primaquine therapy was conducted in consecutive patients admitted to Kapit Hospital, Sarawak, Malaysian Borneo with PCR-confirmed single P. knowlesi infections. These patients were given oral chloroquine for three days, and at 24 hours oral primaquine was administered for two consecutive days, primarily as a gametocidal agent. Clinical and parasitological responses were recorded at 6-hourly intervals during the first 24 hours, daily until discharge and then weekly to day 28. Vivax malaria patients were studied as a comparator group. RESULTS: Of 96 knowlesi malaria patients who met the study criteria, 73 were recruited to an assessment of the acute response to treatment and 60 completed follow-up over 28 days. On admission, the mean parasite stage distributions were 49.5%, 41.5%, 4.0% and 5.6% for early trophozoites, late trophozoites, schizonts and gametocytes respectively. The median fever clearance time was 26.5 [inter-quartile range 16-34] hours. The mean times to 50% (PCT50) and 90% (PCT90) parasite clearance were 3.1 (95% confidence intervals [CI] 2.8-3.4) hours and 10.3 (9.4-11.4) hours. These were more rapid than in a group of 23 patients with vivax malaria 6.3 (5.3-7.8) hours and 20.9 (17.6-25.9) hours; P = 0.02). It was difficult to assess the effect of primaquine on P. knowlesi parasites, due to the rapid anti-malarial properties of chloroquine and since primaquine was administered 24 hours after chloroquine. No P. knowlesi recrudescences or re-infections were detected by PCR. CONCLUSIONS: Chloroquine plus primaqine is an inexpensive and highly effective treatment for uncomplicated knowlesi malaria infections in humans and there is no evidence of drug resistance. Further studies using alternative anti-malarial drugs, including artemisinin derivatives, would be desirable to define optimal management strategies for P. knowlesi

Molecular epidemiology and population genomics of Plasmodium knowlesi.

Molecular epidemiology has been central to uncovering P. knowlesi as an important cause of human malaria in Southeast Asia, and to understanding the complex nature of this zoonosis. Species-specific parasite detection and characterization of sequences were vital to show that P. knowlesi was distinct from the human parasite species that had been presumed to cause all malaria. With established sensitive and specific molecular detection tools, surveys subsequently indicated the distribution of P. knowlesi infections in humans, wild primate reservoir host species, and mosquito vector species. The importance of studying P. knowlesi genetic polymorphism was indicated initially by analysing a few nuclear gene loci as well as the mitochondrial genome, and subsequently by multi-locus microsatellite analyses and whole-genome sequencing. Different human infections generally have unrelated P. knowlesi genotypes, acquired from the diverse local parasite reservoirs in macaques. However, individual human infections are usually less genetically complex than those of wild macaques which experience more frequent superinfection with different P. knowlesi genotypes. Multi-locus analyses have revealed deep population subdivisions within P. knowlesi, which are structured both geographically and in relation to different macaque reservoir host species. Simplified genotypic discrimination assays now enable efficient large-scale surveillance of the sympatric P. knowlesi subpopulations within Malaysian Borneo. The whole-genome sequence analyses have also identified loci under recent positive natural selection in the P. knowlesi genome, with evidence that different loci are affected in different populations. These provide a foundation to understand recent adaptation of the zoonotic parasite populations, and to track and interpret future changes as they emerge

Admixture in Humans of Two Divergent Plasmodium knowlesi Populations Associated with Different Macaque Host Species.

Human malaria parasite species were originally acquired from other primate hosts and subsequently became endemic, then spread throughout large parts of the world. A major zoonosis is now occurring with Plasmodium knowlesi from macaques in Southeast Asia, with a recent acceleration in numbers of reported cases particularly in Malaysia. To investigate the parasite population genetics, we developed sensitive and species-specific microsatellite genotyping protocols and applied these to analysis of samples from 10 sites covering a range of >1,600 km within which most cases have occurred. Genotypic analyses of 599 P. knowlesi infections (552 in humans and 47 in wild macaques) at 10 highly polymorphic loci provide radical new insights on the emergence. Parasites from sympatric long-tailed macaques (Macaca fascicularis) and pig-tailed macaques (M. nemestrina) were very highly differentiated (FST = 0.22, and K-means clustering confirmed two host-associated subpopulations). Approximately two thirds of human P. knowlesi infections were of the long-tailed macaque type (Cluster 1), and one third were of the pig-tailed-macaque type (Cluster 2), with relative proportions varying across the different sites. Among the samples from humans, there was significant indication of genetic isolation by geographical distance overall and within Cluster 1 alone. Across the different sites, the level of multi-locus linkage disequilibrium correlated with the degree of local admixture of the two different clusters. The widespread occurrence of both types of P. knowlesi in humans enhances the potential for parasite adaptation in this zoonotic system

Single Step Solution Processed GaAs Thin Films from GaMe 3 and BuAsH 2 under Ambient Pressure

This article reports on the possibility of low-cost GaAs formed under ambient pressure via a single step solution processed route from only readily available precursors, tBuAsH2 and GaMe3. The thin films of GaAs on glass substrates were found to have good crystallinity with crystallites as large as 150 nm and low contamination with experimental results matching well with theoretical density of states calculations. These results open up a route to efficient and cost-effective scale up of GaAs thin films with high material properties for widespread industrial use. Confirmation of film quality was determined using XRD, Raman, EDX mapping, SEM, HRTEM, XPS, and SIMS

Forehead EEG in Support of Future Feasible Personal Healthcare Solutions: Sleep Management, Headache Prevention, and Depression Treatment

© 2013 IEEE. There are current limitations in the recording technologies for measuring EEG activity in clinical and experimental applications. Acquisition systems involving wet electrodes are time-consuming and uncomfortable for the user. Furthermore, dehydration of the gel affects the quality of the acquired data and reliability of long-term monitoring. As a result, dry electrodes may be used to facilitate the transition from neuroscience research or clinical practice to real-life applications. EEG signals can be easily obtained using dry electrodes on the forehead, which provides extensive information concerning various cognitive dysfunctions and disorders. This paper presents the usefulness of the forehead EEG with advanced sensing technology and signal processing algorithms to support people with healthcare needs, such as monitoring sleep, predicting headaches, and treating depression. The proposed system for evaluating sleep quality is capable of identifying five sleep stages to track nightly sleep patterns. Additionally, people with episodic migraines can be notified of an imminent migraine headache hours in advance through monitoring forehead EEG dynamics. The depression treatment screening system can predict the efficacy of rapid antidepressant agents. It is evident that frontal EEG activity is critically involved in sleep management, headache prevention, and depression treatment. The use of dry electrodes on the forehead allows for easy and rapid monitoring on an everyday basis. The advances in EEG recording and analysis ensure a promising future in support of personal healthcare solutions

Efficient Surveillance of Plasmodium knowlesi Genetic Subpopulations, Malaysian Borneo, 2000-2018.

Population genetic analysis revealed that Plasmodium knowlesi infections in Malaysian Borneo are caused by 2 divergent parasites associated with long-tailed (cluster 1) and pig-tailed (cluster 2) macaques. Because the transmission ecology is likely to differ for each macaque species, we developed a simple genotyping PCR to efficiently distinguish between and survey the 2 parasite subpopulations. This assay confirmed differences in the relative proportions in areas of Kapit division of Sarawak state, consistent with multilocus microsatellite analyses. Analyses of 1,204 human infections at Kapit Hospital showed that cluster 1 caused approximately two thirds of cases with no significant temporal changes from 2000 to 2018. We observed an apparent increase in overall numbers in the most recent 2 years studied, driven mainly by increased cluster 1 parasite infections. Continued monitoring of the frequency of different parasite subpopulations and correlation with environmental alterations are necessary to determine whether the epidemiology will change substantially

Clear cell odontogenic carcinoma: a diagnostic and therapeutic dilemma

BACKGROUND: Clear cell odontogenic carcinoma is a rare odontogenic tumor occurring in the anterior region of the mandible in 5(th)–7(th )decades and shows a female preponderance. It is potentially aggressive, capable of frequent recurrences and loco-regional and distant metastases. CASE PRESENTATION: A 45- year- old woman presented with a radiolucent left mandibular swelling associated with loss of teeth. Left cervical lymph nodes were enlarged on palpation. The patient underwent resection of the tumor but consequent to resected margins being positive for tumor cells underwent left hemimandibulectomy with ipsilateral functional neck dissection and was free of recurrence at 8 months follow-up. CONCLUSION: Clear cell odontogenic carcinoma should be considered in the differential diagnosis of jaw tumors with conspicuous clear cell component. Curettage or conservative resection inevitably results in recurrences and/or metastasis and more radical resection is warranted in these tumors, especially when they are large and show soft tissue invasion