[Molecular dissection of Mycobacterium tuberculosis-containing phagosomes].

Mycobacterium tuberculosis is an intracellular bacterium that can proliferate within phagocytosed macrophages. M. tuberculosis gains this ability by inhibiting phagolysosome biogenesis.

Performance evaluation of the digital cell imaging analyzer DI-60 integrated into the fully automated Sysmex XN hematology analyzer system.

Background: The XN-Series (Sysmex, Kobe, Japan) have been equipped with the automated digital cell imaging analyzer DI-60, which provides complete automation of the sample processing with automated complete blood counts (CBC), slide making/staining, and digital scanning with cell pre-classification. The aim of this study was to evaluate the efficacy of the XN-Series as an integrated blood cell analysis system.

Methods: White blood cell (WBC) morphological analysis by the DI-60 was evaluated using 232 blood samples from patients.

Performance comparison of second- and third-generation sequencers using a bacterial genome with two chromosomes.

Background: The availability of diverse second- and third-generation sequencing technologies enables the rapid determination of the sequences of bacterial genomes. However, identifying the sequencing technology most suitable for producing a finished genome with multiple chromosomes remains a challenge. We evaluated the abilities of the following three second-generation sequencers: Roche 454 GS Junior (GS Jr), Life Technologies Ion PGM (Ion PGM), and Illumina MiSeq (MiSeq) and a third-generation sequencer, the Pacific Biosciences RS sequencer (PacBio), by sequencing and assembling the genome of Vibrio parahaemolyticus, which consists of a 5-Mb genome comprising two circular chromosomes.

Optical coherence tomographic reflectivity of cystoid spaces is related to recurrent diabetic macular edema after triamcinolone.

Purpose: To determine the association between reflectivity levels in foveal cystoid spaces on optical coherence tomography images and the course of the logarithm of the minimum angle of resolution (logMAR) after triamcinolone acetonide treatment for diabetic macular edema.

Methods: We retrospectively reviewed 58 consecutive eyes treated with an intravitreal or sub-Tenon's injection of triamcinolone acetonide for diabetic macular edema. The foveal cystoid spaces within central 1 mm were circumscribed, followed by measurement of optical coherence tomography reflectivity.

Rapid formation of cell-particle complexes via dielectrophoretic manipulation for the detection of surface antigens.

A rapid and simple method for the fabrication of the island patterns with particles and cells was applied to detect the presence of specific antigens on the cell surface. An upper interdigitated microband array (IDA) electrode was mounted on a lower substrate with the same design to fabricate a microfluidic-channel device for dielectrophoretic manipulation. The electrode grid structure was fabricated by rotating the upper template IDA by 90° relative to the lower IDA.

Protective epitopes of the Plasmodium falciparum SERA5 malaria vaccine reside in intrinsically unstructured N-terminal repetitive sequences.

The malaria vaccine candidate antigen, SE36, is based on the N-terminal 47 kDa domain of Plasmodium falciparum serine repeat antigen 5 (SERA5). In epidemiological studies, we have previously shown the inhibitory effects of SE36 specific antibodies on in vitro parasite growth and the negative correlation between antibody level and malaria symptoms. A phase 1 b trial of the BK-SE36 vaccine in Uganda elicited 72% protective efficacy against symptomatic malaria in children aged 6-20 years during the follow-up period 130-365 days post-second vaccination.

Decisions for the future.

Music and science have always been my passion. Ultimately, though, the scale was tipped in favor of science. And I lived out my science dreams in trying to grasp as many facets of it as possible.

Olfactory plays a key role in spatiotemporal pathogenesis of cerebral malaria.

Cerebral malaria is a complication of Plasmodium falciparum infection characterized by sudden coma, death, or neurodisability. Studies using a mouse model of experimental cerebral malaria (ECM) have indicated that blood-brain barrier disruption and CD8 T cell recruitment contribute to disease, but the spatiotemporal mechanisms are poorly understood. We show by ultra-high-field MRI and multiphoton microscopy that the olfactory bulb is physically and functionally damaged (loss of smell) by Plasmodium parasites during ECM.

Z-100, an immunomodulatory extract of Mycobacterium tuberculosis strain Aoyama B, prevents spontaneous lymphatic metastasis of B16-BL6 melanoma.

Lymphatic metastasis is common in advanced-stage carcinoma and is associated with a poor prognosis. However, few effective treatments to inhibit it are available. Z-100 is an immunomodulatory extract of Mycobacterium tuberculosis strain Aoyama B that contains polysaccharides such as arabinomannan and mannan.

Parallelism for quantitative image analysis of photoreceptor-retinal pigment epithelium complex alterations in diabetic macular edema.

Purpose: To propose a new method to quantitatively and comprehensively evaluate photoreceptor-RPE complex alterations on spectral-domain optical coherence tomography (SD-OCT) imaging in eyes with diabetic macular edema (DME).

Methods: Spectral-domain OCT images from a consecutive series of 90 eyes in 79 patients with DME and 30 healthy eyes in 30 volunteers were analyzed retrospectively. The subfoveal area covering the photoreceptor layers was skeletonized and the orientation of the segmented lines was termed "parallelism," which reflects image complexity.

Hematological and biochemical data obtained in rural northern Uganda.

Reference intervals for common hematological and clinical chemistry parameters constitute an important basis for health care. Moreover, with increasing priority in drug and vaccine development for infectious diseases in Africa, the first priority is the safety evaluation and tolerability of the candidate interventions in healthy populations. To accurately assess health status and address adverse events, clinical reference intervals in the target population are necessary.

Validation of microinjection methods for generating knockout mice by CRISPR/Cas-mediated genome engineering.

The CRISPR/Cas system, in which the Cas9 endonuclease and a guide RNA complementary to the target are sufficient for RNA-guided cleavage of the target DNA, is a powerful new approach recently developed for targeted gene disruption in various animal models. However, there is little verification of microinjection methods for generating knockout mice using this approach. Here, we report the verification of microinjection methods of the CRISPR/Cas system.

Generation of rodent malaria parasites with a high mutation rate by destructing proofreading activity of DNA polymerase δ.

Plasmodium falciparum malaria imposes a serious public health concern throughout the tropics. Although genetic tools are principally important to fully investigate malaria parasites, currently available forward and reverse tools are fairly limited. It is expected that parasites with a high mutation rate can readily acquire novel phenotypes/traits; however, they remain an untapped tool for malaria biology.

Integrative understanding of macular morphologic patterns in diabetic retinopathy based on self-organizing map.

Purpose: To integrate parameters on spectral-domain optical coherence tomography (SD-OCT) in diabetic retinopathy (DR) based on the self-organizing map and objectively describe the macular morphologic patterns.

Methods: A total of 336 consecutive eyes of 216 patients with DR for whom clear SD-OCT images were available were retrospectively reviewed. Eleven OCT parameters and the logarithm of the minimal angle of resolution (logMAR) were measured.

Association between cystoid spaces on indocyanine green hyperfluorescence and optical coherence tomography after vitrectomy for diabetic macular oedema.

Purpose: To study retrospectively the characteristics of residual indocyanine green (ICG) fluorescence after ICG-assisted vitrectomy and the association with spectral-domain optical coherence tomography (SD-OCT) findings in diabetic macular oedema (DMO).

Methods: Thirteen consecutive eyes of 12 patients for whom fundus near-infrared fluorescence and 20° retinal sectional images were obtained using HRA2 and Spectralis OCT, respectively, 5 days after vitrectomy combined with ICG-assisted inner limiting membrane peeling for DMO. The relationship between the characteristics of the ICG hyperfluorescence and the cystoid spaces in the outer plexiform layer (OPL) on SD-OCT images was evaluated.

Qualitative and quantitative characteristics of near-infrared autofluorescence in diabetic macular edema.

Objective: To study the characteristics of near-infrared autofluorescence (NIR-AF) imaging and its association with spectral-domain optical coherence tomography (SD-OCT) findings and logarithm of the minimal angle of resolution (logMAR) visual acuity (VA) in diabetic macular edema (DME).

Design: Retrospective, observational, cross-sectional study.

Participants: One hundred twenty-one consecutive eyes of 87 patients with center-involved DME for whom NIR-AF and SD-OCT images of sufficient quality were obtained.

Paternal allele influences high fat diet-induced obesity.

C57BL/6J (B6) mice are susceptible to high-fat diet (HFD)-induced obesity and have been used in metabolism research for many decades. However, the genetic component of HFD-induced obesity has not yet been elucidated. This study reports evidence for a paternal transmission of HFD-induced obesity and a correlated expression of Igf2 and Peg3 (paternal expressed gene 3) imprinted genes.

Performance evaluation of the automated morphological analysis of erythrocytes by CellaVision DM96.

Background: Automated digital morphology systems are utilized for blood cell morphological examination. The aim of this study is to evaluate the accuracy and efficacy of RBC morphological anomaly screening using the CellaVision DM96 (DM96) automated image analysis system.

Methods: The automated analysis of RBC shape, size, and chromasia abnormalities was conducted on the DM96 using 478 blood samples.

Measurement of retinal nerve fiber layer thickness in eyes with optic disc swelling by using scanning laser polarimetry and optical coherence tomography.

Background: The retinal nerve fiber layer thickness (RNFLT) in patients with optic disc swelling of different etiologies was compared using scanning laser polarimetry (SLP) and spectral-domain optical coherence tomography (OCT).

Methods: Forty-seven patients with optic disc swelling participated in the cross-sectional study. Both GDx SLP (enhanced corneal compensation) and Spectralis spectral-domain OCT measurements of RNFLT were made in 19 eyes with papilledema (PE), ten eyes with optic neuritis (ON), and 18 eyes with nonarteritic anterior ischemic optic neuropathy (NAION) at the neuro-ophthalmology clinic at Kyoto University Hospital.

Autophagy adaptor protein p62/SQSTM1 and autophagy-related gene Atg5 mediate autophagosome formation in response to Mycobacterium tuberculosis infection in dendritic cells.

Mycobacterium tuberculosis is an intracellular pathogen that can survive within phagocytic cells by inhibiting phagolysosome biogenesis. However, host cells can control the intracellular M. tuberculosis burden by the induction of autophagy.

Rab39a interacts with phosphatidylinositol 3-kinase and negatively regulates autophagy induced by lipopolysaccharide stimulation in macrophages.

Rab39a has pleiotropic functions in phagosome maturation, inflammatory activation and neuritogenesis. Here, we characterized Rab39a function in membrane trafficking of phagocytosis and autophagy induction in macrophages. Rab39a localized to the periphery of LAMP2-positive vesicles and showed the similar kinetics on the phagosome to that of LAMP1.

Difficulty in locking head screw removal.

Background: Locking plates are an internal fixation material useful in the treatment of bone fractures, which provides effective stabilization between the plate and locking head screws (LHSs) via the locking mechanism. However, difficulty in removing LHSs is relatively common, and such cases can require long surgical procedures or use of special removal equipment. The purpose of this study was to report the incidence and risk factors for difficult removal of LHSs.