Although malaria contributes to a significant public health burden, malaria diagnosis relies heavily on either non-specific clinical symptoms or blood smear microscopy methods developed in the 1930s. These approaches severely misrepresent the number of infected individuals and the reservoir of parasites in malaria-endemic communities and undermine efforts to control disease. Limitations of conventional microscopy-based diagnosis center on time required to examine slides, time required to attain expertise sufficient to diagnose infection accurately, and attrition from the limited number of existing malaria microscopy experts. Earlier studies described magnetic properties of Plasmodium falciparum but did not refine methods to diagnosis infection by all four human malaria parasite species. Here, following specific technical procedures, we show that it is possible to concentrate all four human malaria parasite species, at least 40-fold, on microscope slides using very inexpensive magnets through an approach termed magnetic deposition microscopy. This approach delivered greater sensitivity than a thick smear preparation while maintaining the clarity of a thin smear to simplify species-specific diagnosis. Because the magnetic force necessary to concentrate parasites on the slide is focused at a precise position relative to the magnet surface, it is possible to examine a specific region of the slide for parasitized cells and avoid the time-consuming process of scanning the entire slide surface. These results provide insight regarding new strategies for performing malaria blood smear microscopy.
Introduction: There is immense dispute as to whether perturbations in skeletal muscle mitochondrial function contribute to the onset and progression of type 2 diabetes (T2D). The purpose of this study was to examine differences in mitochondrial dynamics, structure, and energetic function in humans across the spectrum of insulin sensitivity. Methods: 58 sedentary adults (37±12 years) were enrolled into one of three groups based upon the following criteria: (1) healthy weight without T2D (HW; BMI: 22.6±1.8 kg/m2 and HbA1c: 5.0±1.1 %); (2) overweight/obesity without T2D (Ov/Ob; BMI: 32.6±3.3 kg/m2 and HbA1c: 5.6±0.4 %); or (3) overweight/obesity with T2D (T2D; BMI: 36.1±6.5 kg/m2 and HbA1c: 6.9±0.9 %). Participants underwent a 3-day inpatient stay consisting of body composition (DXA), aerobic capacity (VO2MAX), and insulin sensitivity (hyperinsulinemic euglycemic clamp). Prior to insulin sensitivity testing, a skeletal muscle biopsy was obtained for determination of mitochondrial dynamics (Western blot), DNA content (qPCR), ultrastructure (electron microscopy), and respiratory capacity (respirometry). Comparisons were made using a one-way ANOVA with contrasts. Results: Insulin sensitivity and aerobic capacity were lower in Ov/Ob and T2D compared to HW. Markers of mitochondrial fission were higher in T2D (Drp1Ser616, MiD49), and fusion lower in T2D and Ov/Ob (Mfn2). The mitophagy marker Parkin was higher in T2D only while Pink1 was lower in T2D relative to Ov/Ob but not HW. The autophagy marker LC3II was higher in Ov/Ob. Mitochondrial content was lower in Ov/Ob and T2D relative to HW. Mitochondrial respiratory capacity was similar between groups. Conclusions: T2D is associated with heightened expression of proteins required for mitochondrial quality control and reduced mitochondrial volume despite intact respiratory function. These data support the notion that mitochondria adapt to progressive insulin resistance by increasing fragmentation to maintain bioenergetic capacity. Disclosure C. L. Axelrod: None. C. Fealy: Employee; Mission Therapeutics. C. L. Hoppel: None. J. P. Kirwan: None. W. S. Dantas: None. E. R. M. Zunica: None. K. Belmont: None. E. C. Heintz: None. G. Davuluri: None. J. T. Mey: None. M. Erickson: None. H. Fujioka: None. Funding National Institutes of Health (DK108089, GM104940)
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