Explainability of three-dimensional convolutional neural networks for functional magnetic resonance imaging of Alzheimer's disease classification based on gradient-weighted class activation mapping.

Currently, numerous studies focus on employing fMRI-based deep neural networks to diagnose neurological disorders such as Alzheimer's Disease (AD), yet only a handful have provided results regarding explainability. We address this gap by applying several prevalent explainability methods such as gradient-weighted class activation mapping (Grad-CAM) to an fMRI-based 3D-VGG16 network for AD diagnosis to improve the model's explainability. The aim is to explore the specific Region of Interest (ROI) of brain the model primarily focuses on when making predictions, as well as whether there are differences in these ROIs between AD and normal controls (NCs). First, we utilized multiple resting-state functional activity maps including ALFF, fALFF, ReHo, and VMHC to reduce the complexity of fMRI data, which differed from many studies that utilized raw fMRI data. Compared to methods utilizing raw fMRI data, this manual feature extraction approach may potentially alleviate the model's burden. Subsequently, 3D-VGG16 were employed for AD classification, where the final fully connected layers were replaced with a Global Average Pooling (GAP) layer, aimed at mitigating overfitting while preserving spatial information within the feature maps. The model achieved a maximum of 96.4% accuracy on the test set. Finally, several 3D CAM methods were employed to interpret the models. In the explainability results of the models with relatively high accuracy, the highlighted ROIs were primarily located in the precuneus and the hippocampus for AD subjects, while the models focused on the entire brain for NC. This supports current research on ROIs involved in AD. We believe that explaining deep learning models would not only provide support for existing research on brain disorders, but also offer important referential recommendations for the study of currently unknown etiologies.

Brain-like illusion produced by Skye's Oblique Grating in deep neural networks.

The analogy between the brain and deep neural networks (DNNs) has sparked interest in neuroscience. Although DNNs have limitations, they remain valuable for modeling specific brain characteristics. This study used Skye's Oblique Grating illusion to assess DNNs' relevance to brain neural networks. We collected data on human perceptual responses to a series of visual illusions. This data was then used to assess how DNN responses to these illusions paralleled or differed from human behavior. We performed two analyses:(1) We trained DNNs to perform horizontal vs. non-horizontal classification on images with bars tilted different degrees (non-illusory images) and tested them on images with horizontal bars with different illusory strengths measured by human behavior (illusory images), finding that DNNs showed human-like illusions; (2) We performed representational similarity analysis to assess whether illusory representation existed in different layers within DNNs, finding that DNNs showed illusion-like responses to illusory images. The representational similarity between real tilted images and illusory images was calculated, which showed the highest values in the early layers and decreased layer-by-layer. Our findings suggest that DNNs could serve as potential models for explaining the mechanism of visual illusions in human brain, particularly those that may originate in early visual areas like the primary visual cortex (V1). While promising, further research is necessary to understand the nuanced differences between DNNs and human visual pathways.

Uniformly positive or negative correlation of cerebral gray matter regions with driving safety behaviors of healthy older drivers.

This study investigated the relationship between cerebral gray matter (GM) regions and driving safety behaviors (DSBs) of 98 older drivers without dementia (mean age, 77.72 ± 3.677 years). Their DSBs were evaluated on actual vehicles running on a closed-circuit course. The DSB was scored in six categories: DSB1, visual search behavior; DSB2, speeding; DSB3, signaling of the indicator; DSB4, vehicle stability; DSB5, positioning; and DSB6, steering. The scores were calculated by a single driving instructor; larger scores indicated safer driving performances. Regional GM volumes were measured with voxel-based morphometry by magnetic resonance imaging (MRI). Out of 56 GM regions, 18 were correlated with DSB categories except for DSB4. When a single GM region was correlated with multiple DSB categories, a positive or negative response was uniformly determined for the respective region despite clear differences in the DSB categories. This result suggests the possible existence of two contradictory mechanisms in the brain for DSB. The left postcentral gyrus may largely function in regulating DSBs because it was negatively correlated with five of six DSB categories. Thus, MRI's measurement of regional GM volumes may help deepen the understanding of the diversity and complexity inherent in brain functions for DSBs.

Protective role of stratum corneum in percutaneous Leptospira infection in a hamster model.

Leptospira enters humans and animals through injured skin or mucous membranes by direct or indirect contact with urine excreted from infected reservoirs. Individuals with cut or scratched skin are at high risk of infection and are recommended to be protected from contact with Leptospira, but the risk of infection via skin without apparent wounds is unknown. We hypothesized that the stratum corneum of the epidermis might prevent percutaneous invasion of leptospires. We established a stratum corneum deficient model of hamsters using the tape stripping method. The mortality rate of hamsters lacking stratum corneum that were exposed to Leptospira was higher than that of controls with shaved skin, and was not significantly different from an epidermal wound group. These results indicated that the stratum corneum plays a critical role in protecting the host against leptospiral entry. We also examined the migration of leptospires through the monolayer of HaCaT cells (human keratinocyte cell line) using Transwell. The number of pathogenic leptospires penetrating the HaCaT cell monolayers was higher than that of non-pathogenic leptospires. Furthermore, scanning and transmission electron microscopic observations revealed that the bacteria penetrated the cell monolayers through both intracellular and intercellular routes. This suggested that pathogenic Leptospira can migrate easily through keratinocyte layers and is associated with virulence. Our study further highlights the importance of the stratum corneum as a critical barrier against the invasion of Leptospira found in contaminated soil and water. Hence, preventative measures against contact infection should be taken, even without visible skin wounds.

Artificial Intelligence in Quantitative Chest Imaging Analysis for Occupational Lung Disease.

Occupational lung disease manifests complex radiologic findings which have long been a challenge for computer-assisted diagnosis (CAD). This journey started in the 1970s when texture analysis was developed and applied to diffuse lung disease. Pneumoconiosis appears on radiography as a combination of small opacities, large opacities, and pleural shadows. The International Labor Organization International Classification of Radiograph of Pneumoconioses has been the main tool used to describe pneumoconioses and is an ideal system that can be adapted for CAD using artificial intelligence (AI). AI includes machine learning which utilizes deep learning or an artificial neural network. This in turn includes a convolutional neural network. The tasks of CAD are systematically described as classification, detection, and segmentation of the target lesions. Alex-net, VGG16, and U-Net are among the most common algorithms used in the development of systems for the diagnosis of diffuse lung disease, including occupational lung disease. We describe the long journey in the pursuit of CAD of pneumoconioses including our recent proposal of a new expert system.

Primary grade 2 neuroendocrine tumor of the ileal mesentery: a case report.

BACKGROUND: The prevalence and incidence of neuroendocrine tumors (NETs) are increasing worldwide. Primary mesenteric NETs are extremely rare. Solid tumors that arise in the mesentery are typically metastatic. We present an extremely rare case of a primary grade 2 NET (NET G2) in the ileal mesentery. CASE PRESENTATION: A 54-year-old man was referred to our hospital for further examination of a previously diagnosed right mesenteric tumor. Mild tenderness was noted on the right side of the abdomen, but there were no palpable masses. Fluorodeoxyglucose-positron emission tomography (FDG-PET) revealed slight FDG uptake (maximum standardized uptake value, 2.0) in the right abdomen, and a benign or low-grade malignant tumor was suspected. We extracted the ileal mesenteric tumor with an ileal resection (90 cm). The cut surface of the 55 × 33 × 33 mm3 tumor was pale yellowish-white. Immunohistochemistry revealed diffuse staining for synaptophysin and chromogranin A, and focal staining for CD56. The Ki-67 index was 3%. The final pathological diagnosis was NET G2. The patient's postoperative course was uneventful, and he developed no recurrence 1.5 years after surgery. Postoperative antitumor therapy was not performed for this patient because the histological diagnosis was NET G2, and it was determined that the tumor could be completely resected by surgery. CONCLUSIONS: We report an extremely rare case of primary ileal mesenteric NET. Mesenteric tumors that show slight FDG uptake on FDG-PET examination should be considered well-differentiated NET.

Leptospira Is an Environmental Bacterium That Grows in Waterlogged Soil.

Leptospirosis is a zoonotic disease caused by infection with pathogenic leptospires. Consistent with recent studies by other groups, leptospires were isolated from 89 out of 110 (80.9%) soil or water samples from varied locations in the Philippines in our surveillance study, indicating that leptospires might have a life cycle that does not involve animal hosts. However, despite previous work, it has not been confirmed whether leptospires multiply in the soil environment under various experimental conditions. Given the fact that the case number of leptospirosis is increased after flood, we hypothesized that waterlogged soil, which mimics the postflooding environment, could be a suitable condition for growing leptospires. To verify this hypothesis, pathogenic and saprophytic leptospires were seeded in the bottles containing 2.5 times as much water as soil, and bacterial counts in the bottles were measured over time. Pathogenic and saprophytic leptospires were found to increase their number in waterlogged soil but not in water or soil alone. In addition, leptospires were reisolated from soil in closed tubes for as long as 379 days. These results indicate that leptospires are in a resting state in the soil and are able to proliferate with increased water content in the environment. This notion is strongly supported by observations that the case number of leptospirosis is significantly higher in rainy seasons and increased after flood. Therefore, we reached the following conclusion: environmental soil is a potential reservoir of leptospires. IMPORTANCE Since research on Leptospira has focused on pathogenic leptospires, which are supposed to multiply only in animal hosts, the life cycle of saprophytic leptospires has long been a mystery. This study demonstrates that both pathogenic and saprophytic leptospires multiply in the waterlogged soil, which mimics the postflooding environment. The present results potentially explain why leptospirosis frequently occurs after floods. Therefore, environmental soil is a potential reservoir of leptospires and leptospirosis is considered an environment-borne as well as a zoonotic disease. This is a significant report to reveal that leptospires multiply under environmental conditions, and this finding leads us to reconsider the ecology of leptospires.

Novel Virulent Bacteriophage ΦSG005, Which Infects Streptococcus gordonii, Forms a Distinct Clade among Streptococcus Viruses.

Bacteriophages are viruses that specifically infect bacteria and are classified as either virulent phages or temperate phages. Despite virulent phages being promising antimicrobial agents due to their bactericidal effects, the implementation of phage therapy depends on the availability of virulent phages against target bacteria. Notably, virulent phages of Streptococcus gordonii, which resides in the oral cavity and is an opportunistic pathogen that can cause periodontitis and endocarditis have previously never been found. We thus attempted to isolate virulent phages against S. gordonii. In the present study, we report for the first time a virulent bacteriophage against S. gordonii, ΦSG005, discovered from drainage water. ΦSG005 is composed of a short, non-contractile tail and a long head, revealing Podoviridae characteristics via electron microscopic analysis. In turbidity reduction assays, ΦSG005 showed efficient bactericidal effects on S. gordonii. Whole-genome sequencing showed that the virus has a DNA genome of 16,127 bp with 21 coding sequences. We identified no prophage-related elements such as integrase in the ΦSG005 genome, demonstrating that the virus is a virulent phage. Phylogenetic analysis indicated that ΦSG005 forms a distinct clade among the streptococcus viruses and is positioned next to streptococcus virus C1. Molecular characterization revealed the presence of an anti-CRISPR (Acr) IIA5-like protein in the ΦSG005 genome. These findings facilitate our understanding of streptococcus viruses and advance the development of phage therapy against S. gordonii infection.

3D MRI Reconstruction Based on 2D Generative Adversarial Network Super-Resolution.

The diagnosis of brain pathologies usually involves imaging to analyze the condition of the brain. Magnetic resonance imaging (MRI) technology is widely used in brain disorder diagnosis. The image quality of MRI depends on the magnetostatic field strength and scanning time. Scanners with lower field strengths have the disadvantages of a low resolution and high imaging cost, and scanning takes a long time. The traditional super-resolution reconstruction method based on MRI generally states an optimization problem in terms of prior information. It solves the problem using an iterative approach with a large time cost. Many methods based on deep learning have emerged to replace traditional methods. MRI super-resolution technology based on deep learning can effectively improve MRI resolution through a three-dimensional convolutional neural network; however, the training costs are relatively high. In this paper, we propose the use of two-dimensional super-resolution technology for the super-resolution reconstruction of MRI images. In the first reconstruction, we choose a scale factor of 2 and simulate half the volume of MRI slices as input. We utilize a receiving field block enhanced super-resolution generative adversarial network (RFB-ESRGAN), which is superior to other super-resolution technologies in terms of texture and frequency information. We then rebuild the super-resolution reconstructed slices in the MRI. In the second reconstruction, the image after the first reconstruction is composed of only half of the slices, and there are still missing values. In our previous work, we adopted the traditional interpolation method, and there was still a gap in the visual effect of the reconstructed images. Therefore, we propose a noise-based super-resolution network (nESRGAN). The noise addition to the network can provide additional texture restoration possibilities. We use nESRGAN to further restore MRI resolution and high-frequency information. Finally, we achieve the 3D reconstruction of brain MRI images through two super-resolution reconstructions. Our proposed method is superior to 3D super-resolution technology based on deep learning in terms of perception range and image quality evaluation standards.

Engineering of Aeromonas caviae Polyhydroxyalkanoate Synthase Through Site-Directed Mutagenesis for Enhanced Polymerization of the 3-Hydroxyhexanoate Unit.

Polyhydroxyalkanoate (PHA) synthase is an enzyme that polymerizes the acyl group of hydroxyacyl-coenzyme A (CoA) substrates. Aeromonas caviae PHA synthase (PhaCAc) is an important biocatalyst for the synthesis of a useful PHA copolymer, poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] [P(3HB-co-3HHx)]. Previously, a PhaCAc mutant with double mutations in asparagine 149 (replaced by serine [N149S]) and aspartate 171 (replaced by glycine [D171G]) was generated to synthesize a 3HHx-rich P(3HB-co-3HHx) and was named PhaCAc NSDG. In this study, to further increase the 3HHx fraction in biosynthesized PHA, PhaCAc was engineered based on the three-dimensional structural information of PHA synthases. First, a homology model of PhaCAc was built to target the residues for site-directed mutagenesis. Three residues, namely tyrosine 318 (Y318), serine 389 (S389), and leucine 436 (L436), were predicted to be involved in substrate recognition by PhaCAc. These PhaCAc NSDG residues were replaced with other amino acids, and the resulting triple mutants were expressed in the engineered strain of Ralstonia eutropha for application in PHA biosynthesis from palm kernel oil. The S389T mutation allowed the synthesis of P(3HB-co-3HHx) with an increased 3HHx fraction without a significant reduction in PHA yield. Thus, a new workhorse enzyme was successfully engineered for the biosynthesis of a higher 3HHx-fraction polymer.

L-Ascorbate Biosynthesis Involves Carbon Skeleton Rearrangement in the Nematode Caenorhabditis elegans.

Ascorbate (AsA) is required as a cofactor and is widely distributed in plants and animals. Recently, it has been suggested that the nematode Caenorhabditis elegans also synthesizes AsA. However, its biosynthetic pathway is still unknown. To further understand AsA biosynthesis in C. elegans, we analyzed the incorporation of the 13C atom into AsA using gas chromatography-mass spectrometry (GC-MS) in worms fed with D-Glc (1-13C)-labeled Escherichia coli. GC-MS analysis revealed that AsA biosynthesis in C. elegans, similarly to that in mammalian systems, involves carbon skeleton rearrangement. The addition of L-gulono-1,4-lactone, an AsA precursor in the mammalian pathway, significantly increased AsA level in C. elegans, whereas the addition of L-galactono-1,4-lactone, an AsA precursor in the plant and Euglena pathway, did not affect AsA level. The suppression of E03H4.3 (an ortholog of gluconolactonase) or the deficiency of F54D5.12 (an ortholog of L-gulono-1,4-lactone oxidase) significantly decreased AsA level in C. elegans. Although N2- and AsA-deficient F54D5.12 knockout mutant worm (tm6671) morphologies and the ratio of collagen to non-collagen protein did not show any significant differences, the mutant worms exhibited increased malondialdehyde levels and reduced lifespan compared with the N2 worms. In conclusion, our findings indicate that the AsA biosynthetic pathway is similar in C. elegans and mammals.

Molecular and phenotypic characterization of Leptospira johnsonii sp. nov., Leptospira ellinghausenii sp. nov. and Leptospira ryugenii sp. nov. isolated from soil and water in Japan.

In a previous study, 50 of 132 soil samples collected throughout Japan were found to be Leptospira-positive. In the present study, three strains identified in the collected specimens, three, E8, E18 and YH101, were found to be divergent from previously described Leptospira species according to 16S ribosomal RNA gene sequence analysis. These three strains have a helical shape similar to that of typical Leptospira and were not re-isolated from experimental mice inoculated with the cultured strains. Upon 16S ribosomal RNA gene sequence analysis, E8 was found to belong to the intermediate Leptospira species clade and E18 and YH101 to belong to the saprophytic Leptospira species clade. Based on analyses of genome-to-genome distances and average nucleotide identity in silico using whole genome sequences and DNA-DNA hybridization in vitro, these isolates were found to be distinct from previously described Leptospira species. Therefore, these three isolates represent novel species of the genus Leptospira for which the names Leptospira johnsonii sp. nov., (type strain E8 T , = JCM 32515 T = CIP111620 T ), Leptospira ellinghausenii sp. nov., (type strain E18 T , = JCM 32516 T = CIP111618 T ) and Leptospira ryugenii sp. nov., (type strain YH101 T , = JCM 32518 T = CIP111617 T ) are proposed.

Molecular analysis of Streptococcus pyogenes strains isolated from patients with recurrent pharyngitis after oral amoxicillin treatment.

PURPOSE: The most common illness caused by Streptococcus pyogenes (Group A streptococcus; GAS) is acute pharyngitis. It has been reported that a small percentage of patients experience recurrent GAS pharyngitis after 10 days of treatment with oral amoxicillin. The aim of this study was to clarify whether recurrent GAS pharyngitis is reactivation caused by the primary strain remaining at the infection site, or if the reinfection is caused by newly acquired strains. METHODOLOGY: A total of 135 GAS clinical strains were isolated from the tonsils of 116 pediatric patients with acute GAS pharyngitis between November, 2012 and April, 2014 in Saga, Japan. These strains were analysed by pulsed-field gel electrophoresis (PFGE)-typing methods. RESULTS: The isolates were grouped into 16 PFGE-types. The epidemic PFGE types that caused pharyngitis were found to change dynamically during 18 months. Eleven strains caused recurrent pharyngitis within 40 days after the last treatment, all of them showing the same PFGE-type as the primary strains. Eight of the strains caused recurrence more than 40 days after the treatment. Among them, six showed different PFGE-types from the primary strains. CONCLUSION: When recurrent pharyngitis emerges more than 40 days after the last treatment, penicillin can be prescribed again because reinfection is suspected. However, when recurrent pharyngitis takes place within 40 days after completing the treatment, alternative drugs should be considered for retreatment because the pharyngitis is likely to be due to reactivation.

Characterization of Leptospira species isolated from soil collected in Japan.

Leptospira were isolated from soil obtained from Hokkaido, the northernmost island, to Okinawa, the southernmost island, of Japan using sulfamethoxazole, trimethoprim, amphotericin B, fosfomycin, and 5- fluorouracil. Fifty of 132 soil samples (37.9%) were culture-positive. On the basis of 16S-rDNA sequences, 12 of the isolated Leptospira were classified into a pathogenic species clade that is closely associated with L. alstonii and L. kmetyi. Nine isolates were classified as intermediate species and were found to be similar to L. licerasiae. Twenty-seven isolates were classified as non-pathogenic species, of which 23 were found to be related to L. wolbachii. Non-pathogenic Leptospira are commonly distributed in environmental soil.

Adipose tissue is the first colonization site of Leptospira interrogans in subcutaneously infected hamsters.

Leptospirosis is one of the most widespread zoonoses in the world, and its most severe form in humans, "Weil's disease," may lead to jaundice, hemorrhage, renal failure, pulmonary hemorrhage syndrome, and sometimes,fatal multiple organ failure. Although the mechanisms underlying jaundice in leptospirosis have been gradually unraveled, the pathophysiology and distribution of leptospires during the early stage of infection are not well understood. Therefore, we investigated the hamster leptospirosis model, which is the accepted animal model of human Weil's disease, by using an in vivo imaging system to observe the whole bodies of animals infected with Leptospira interrogans and to identify the colonization and growth sites of the leptospires during the early phase of infection. Hamsters, infected subcutaneously with 104 bioluminescent leptospires, were analyzed by in vivo imaging, organ culture, and microscopy. The results showed that the luminescence from the leptospires spread through each hamster's body sequentially. The luminescence was first detected at the injection site only, and finally spread to the central abdomen, in the liver area. Additionally, the luminescence observed in the adipose tissue was the earliest detectable compared with the other organs, indicating that the leptospires colonized the adipose tissue at the early stage of leptospirosis. Adipose tissue cultures of the leptospires became positive earlier than the blood cultures. Microscopic analysis revealed that the leptospires colonized the inner walls of the blood vessels in the adipose tissue. In conclusion, this is the first study to report that adipose tissue is an important colonization site for leptospires, as demonstrated by microscopy and culture analyses of adipose tissue in the hamster model of Weil's disease.

Polymer-brush-afforded SPIO Nanoparticles Show a Unique Biodistribution and MR Imaging Contrast in Mouse Organs.

INTRODUCTION: To investigate the biodistribution and retention properties of the new super paramagnetic iron oxide (new SPIO: mean hydrodynamic diameter, 100 nm) nanoparticles, which have concentrated polymer brushes in the outer shell and are difficult for phagocytes to absorb, and to compare the new SPIO with clinically approved SPIO (Resovist: mean hydrodynamic diameter, 57 nm). MATERIALS AND METHODS: 16 male C57BL/6N mice were divided in two groups according to the administered SPIO (n = 8 for each group; intravenous injection does, 0.1 ml). In vivo magnetic resonance imaging (MRI) was performed before and one hour, one day, one week and four weeks after SPIO administration by two dimensional-the fast low angle shot (2D-FLASH) sequence at 11.7T. Ex vivo high-resolution images of fixed organs were also obtained by (2D-FLASH). After the ex vivo MRI, organs were sectioned and evaluated histologically to confirm the biodistribution of each particle precisely. RESULTS: The new SPIO was taken up in small amounts by liver Kupffer cells and showed a unique in vivo MRI contrast pattern in the kidneys, where the signal intensity decreased substantially in the boundaries between cortex and outer medulla and between outer and inner medulla. We found many round dark spots in the cortex by ex vivo MRI in both groups. Resovist could be detected almost in the cortex. The shapes of the dark spots were similar to those observed in the new SPIO group. Transmission electron microscopy revealed that Resovist and the new SPIO accumulated in different cells of glomeruli, that is, endothelial and mesangial cells, respectively. CONCLUSION: The new SPIO was taken up in small amounts by liver tissue and showed a unique MRI contrast pattern in the kidney. The SPIO were found in the mesangial cells of renal corpuscles. Our results indicate that the new SPIO may be potentially be used as a new contrast agent for evaluation of kidney function as well as immunune function.

Non-exponential growth of Mycobacterium leprae Thai-53 strain cultured in vitro.

In this study, attempts were made to culture this bacterium in media supplemented with a variety of biological materials to determine why cultivation of Mycobacterium leprae in vitro has not this far been successful. A slight increase in the number of cells in medium supplemented with human blood plasma and an extract of nude mouse tissue as observed after more than 3 months of cultivation at 30 °C. To ascertain whether this increase was real growth, the growth was analyzed by droplet digital PCR, which showed a slow increase in the copy number of cell-associated DNA and the release of a large amount of DNA into the culture medium from bacterial cells during cultivation. These results were supported by electron microscopic examination of M. leprae in infected mouse tissues, which showed that most of the replicated bacteria had degenerated and only a few cells survived. Based on these results, it was postulated that many of the replicated cells degenerate during M. leprae growth and that only a few cells remain to participate in the next growth stage. This means that, unlike other cultivable bacteria, the growth of M. leprae is not exponential and the number of cells therefore increase extremely slowly. Thus, accurate judging of the success of M. leprae cultivation requires observation of growth over a long period of time and careful measurement of the increase in number of viable cells.

A soluble form of Siglec-9 provides a resistance against Group B Streptococcus (GBS) infection in transgenic mice.

Group B Streptococcus (GBS) is a leading cause of invasive bacterial infections in human newborns. A key GBS virulence factor is its capsular polysaccharide (CPS), possessing terminal sialic acid residues that suppress host immune response and provide a survival advantage to the pathogen. CPS binds to Siglec-9 expressed on neutrophils, which is expected to down-regulate the immune responsiveness of neutrophils. We hypothesized that a soluble form of Siglec-9 (sSiglec-9) competitively inhibits a binding of CPS to Siglec-9 on immune cells, leading to provide antibacterial benefit against GBS infection in the transgenic mouse line expressing sSiglec-9 (sSiglec-9 Tg). The sSiglec-9 in the sera of sSiglec-9 Tg bound to the sialylated-GBS strains belonging to serotypes Ia, Ib, II, III, IV and V in whole GBS cell ELISA. When GBS cells of serotype III that is a common serotype in late-onset GBS disease (LOD) were intraperitoneally inoculated into sSiglec-9 Tg, sSiglec-9 Tg showed a significant resistance as compared with non-transgenic littermates. Furthermore, GBS serotype III organisms were not detected in cultures of the blood from surviving mice (<1 × 103 CFU/ml). These results indicated that sSiglec-9 Tg mice were more efficient in eliminating GBS and survived better after the intraperitoneal challenge with GBS serotype III bacteria.

Legionella dumoffii Tex-KL Mutated in an Operon Homologous to traC-traD is Defective in Epithelial Cell Invasion.

OBJECTIVE: To understand the mechanism of invasion by Legionella dumoffii. METHODS: The L. dumoffii strain Tex-KL was mutated using the Tn903 derivative, Tn903dIIlacZ. After screening 799 transposon insertion mutants, we isolated one defective mutant. We then constructed the gene-disrupted mutant, KL16, and studied its invasion of and intracellular growth in HeLa and A549 cells, and in A/J mice survival experiments. The structure of traC-traD operon was analyzed by RT-PCR. RESULTS: The transposon insertion was in a gene homologous to Salmonella typhi traC, which is required for the assembly of F pilin into the mature F pilus structure and for conjugal DNA transmission. Results from RT-PCR suggested that the traC-traD region formed an operon. We found that when the traC gene was disrupted, invasion and intracellular growth of L. dumoffii Tex-KL were impaired in human epithelial cells. When mice were infected by intranasal inoculation with a traC deficient mutant, their survival significantly increased when compared to mice infected with the wild-type strain.. CONCLUSION: Our results indicated that the traC-traD operon is required for the invasion and intracellular growth abilities of L. dumoffii Tex-KL in epithelial cells.

Colony types and virulence traits of Legionella feeleii determined by exopolysaccharide materials.

Legionella feeleii is a Gram-negative pathogenic bacterium that causes Pontiac fever and pneumonia in humans. When L. feeleii serogroup 1 (ATCC 35072) was cultured on BCYE agar plates, two types of colonies were observed and exhibited differences in color, opacity and morphology. Since the two colony types are white rugose and brown translucent, they were termed as white rugose L. feeleii (WRLf) and brown translucent L. feeleii (BTLf), respectively. They exhibited different growth capacities in BYE broth in vitro, and it was found that WRLf could transform to BTLf. Under the electron microscope, it was observed that WRLf secreted materials which could be stained with ruthenium red, which was absent in BTLf. When U937 macrophages and HeLa cells were infected with the bacteria, WRLf manifested stronger internalization ability than BTLf. Intracellular growth in murine macrophages and Acanthamoeba cells was affected by the level of initial phagocytosis. WRLf was more resistant to human serum bactericidal action than BTLf. After being inoculated to guinea pigs, both organisms caused fever in the animals. These results suggest that ruthenium red-stained materials secreted in the surroundings may play a crucial role in determining L. feeleii colony morphology and virulence traits.