Altered B cell immunoglobulin signature exhibits potential diagnostic values in human colorectal cancer.

Antibody-secreting B cells have long been considered the central element of gut homeostasis; however, tumor-associated B cells in human colorectal cancer (CRC) have not been well characterized. Here, we show that the clonotype, phenotype, and immunoglobulin subclasses of tumor-infiltrating B cells have changed compared to adjacent normal tissue B cells. Remarkably, the tumor-associated B cell immunoglobulin signature alteration can also be detected in the plasma of patients with CRC, suggesting that a distinct B cell response was also evoked in CRC. We compared the altered plasma immunoglobulin signature with the existing method of CRC diagnosis. Our diagnostic model exhibits improved sensitivity compared to the traditional biomarkers, CEA and CA19-9. These findings disclose the altered B cell immunoglobulin signature in human CRC and highlight the potential of using the plasma immunoglobulin signature as a non-invasive method for the assessment of CRC.

Biologically Emulated Flexible Sensors With High Sensitivity and Low Hysteresis: Toward Electronic Skin to a Sense of Touch.

Recently, flexible pressure sensors (FPSs) have attracted intensive attention owing to their ability to mimic and function as electronic skin. Some sensors are exploited with a biological structure dielectric layer for high sensitivity and detection. However, traditional sensors with bionic structures usually suffer from a limited range for high-pressure scenes due to their high sensitivity and high hysteresis in the medium pressure range. Here, a reconfigurable flea bionic structure FPS based on 3D printing technology, which can meet the needs of different scenes via tailoring of the dedicated structural parameters, is proposed. FPS exhibits high sensitivity (1.005 kPa-1 in 0-1 kPa), wide detection range (200 kPa), high repeatability (6000 cycles in 10 kPa), low hysteresis (1.3%), fast response time (40 ms), and very low detection limit (0.5 Pa). Aiming at practical application implementation, FPS has been correspondingly placed on a finger, elbow, arm, neck, cheek, and manipulators to detect the actions of various body parts, suggestive of excellent applicability. It is also integrated to make a flexible 3 × 3 sensor array for detecting spatial pressure distribution. The results indicate that FPS exhibits a significant application potential in advanced biological wearable technologies, such as human motion monitoring.

Segmentation of Laser Marks of Diabetic Retinopathy in the Fundus Photographs Using Lightweight U-Net.

Diabetic retinopathy (DR) is a prevalent vision-threatening disease worldwide. Laser marks are the scars left after panretinal photocoagulation, a treatment to prevent patients with severe DR from losing vision. In this study, we develop a deep learning algorithm based on the lightweight U-Net to segment laser marks from the color fundus photos, which could help indicate a stage or providing valuable auxiliary information for the care of DR patients. We prepared our training and testing data, manually annotated by trained and experienced graders from Image Reading Center, Zhongshan Ophthalmic Center, publicly available to fill the vacancy of public image datasets dedicated to the segmentation of laser marks. The lightweight U-Net, along with two postprocessing procedures, achieved an AUC of 0.9824, an optimal sensitivity of 94.16%, and an optimal specificity of 92.82% on the segmentation of laser marks in fundus photographs. With accurate segmentation and high numeric metrics, the lightweight U-Net method showed its reliable performance in automatically segmenting laser marks in fundus photographs, which could help the AI assist the diagnosis of DR in the severe stage.

Heterogeneity of Visual Preferences for Biological and Repetitive Movements in Children With Autism Spectrum Disorder.

Previous studies have repeatedly reported atypical visual preferences to repetitive movements and deficient perception of biological movements in individuals with autism spectrum disorder (ASD). However, limited research has investigated the heterogeneity of the visual preferences in individuals with ASD. In the current study, we explored the visual preferences to different movement types (repetitive, biological, and random) in children with ASD using a paired preferential looking paradigm. Thirty-nine children with ASD and 37 typically developing (TD) children participated in our study, with their eye movements recorded as the index of visual preferences. We examined the differences of visual preferences between the ASD and TD group, and the heterogeneity of visual preferences within the ASD group. We found group differences between children with ASD and TD children: Overall, the ASD group preferred repetitive movements while the TD group preferred biological movements. We also detected heterogeneity of visual preferences within the ASD group: Although the majority of children with ASD preferred repetitive movements as previous studies reported, 9 out of 39 children with ASD preferred biological movements similarly as their TD peers. Moreover, the visual preference patterns were correlated with autistic symptoms, especially the socio-communicative impairments. Our study provided evidence of heterogeneity of visual attention and main visual preference to repetitive movements in children with ASD. The findings add to the body of literature of the heterogeneous behavioral symptoms and the atypical visual preferences in individuals with ASD. LAY SUMMARY: The current study examined visual preferences to biological, repetitive, and random movements in children with Autism Spectrum Disorder (ASD). We showed a pair of two videos representing two types of movements (random, repetitive, or biological movements) to children with ASD and typically developing children. We found the main visual preferences for repetitive movements and heterogeneity of visual attention within the ASD group. Our findings provide theoretical and methodological implications for future study of the heterogeneity in the ASD population.

Impact of Chemotherapy Regimens on Normal Tissue Complication Probability Models of Acute Hematologic Toxicity in Rectal Cancer Patients Receiving Intensity Modulated Radiation Therapy With Concurrent Chemotherapy From a Prospective Phase III Clinical Trial.

Purpose: To determine whether there are differences in bone marrow tolerance to chemoradiotherapy (CRT) between two chemotherapy regimens according to FOWARC protocol and how chemotherapy regimens affect radiation dose parameters and normal tissue complication probability (NTCP) modelings that correlate with acute hematologic toxicity (HT) in rectal cancer patients treated with intensity modulated radiation therapy (IMRT) and concurrent chemotherapy. Materials and Methods: One hundred and twenty-eight rectal cancer patients who received IMRT from a single institution were recruited from Chinese FOWARC multicenter, open-label, randomized phase III trial. We assessed HT in these patients who were separated into two groups: Oxaliplatin (L-OHP) + 5- fluorouracil (5FU) (FOLFOX, 70 of 128) and 5FU (58 of 128). The pelvic bone marrow (PBM) was divided into three subsites: lumbosacral spine (LSS), ilium (I), and lower pelvic (LP). The endpoint for HT was grade ≥3 (HT3+) and grade ≥2 (HT2+) leukopenia, neutropenia, anemia and thrombocytopenia. Logistic regression was used to analyze the association between HT2+/HT3+ and dosimetric parameters. Lyman-Kutcher-Burman (LKB) model was used to calculate NTCP. Results: Sixty-eight patients experienced HT2+: 22 of 58 (37.9%) 5FU and 46 of 70 (65.7%) FOLFOX (p = 0.008), while twenty-six patients experienced HT3+: 4 of 58 (6.9%) 5FU and 22 of 70 (31.4%) FOLFOX (p = 0.016). PBM and LP dosimetric parameters were correlated with HT2+ in the 5FU group but not in the FOLFOX group. No PBM dosimetric parameters were correlated with HT3+ in both groups. For PBM, NTCP at HT3+ was 0.32 in FOLFOX group relative to 0.10 in 5FU subset (p < 0.05). Conclusion: Patients receiving FOLFOX have lower BM tolerance to CRT than those receiving 5FU. Low-dose radiation to the PBM is predictive for HT2+ in patients who received 5FU. NTCP modeling in FOLFOX group predicts much higher risk of HT3+ than 5FU group.

Primary replication and invasion of the bovine gammaherpesvirus BoHV-4 in the genital mucosae.

Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus that is widespread in cattle. Ex vivo models with bovine genital tract mucosa explants were set up to study molecular/cellular BoHV-4-host interactions. Bovine posterior vagina, cervix and uterus body were collected from cows at two stages of the reproductive cycle for making mucosa explants. The BoHV-4 replication kinetics and characteristics within the three different mucosae of animals in the follicular and luteal phase were assessed by virus titration. The number of plaques, plaque latitude and number of infected cells were determined by immunofluorescence. BoHV-4 replicated in a productive way in all genital mucosal tissues. It infected single individual cells in both epithelium and lamina propria of the genital mucosae at 24 hours post-inoculation (hpi). Later, small BoHV-4 epithelial plaques were formed that did not spread through the basement membrane. 50% of the number of BoHV-4 infected cells were identified as cytokeratin+ and CD172a+ cells in the three parts of the genital tract at 24 hpi. Upon a direct injection of genital explants with BoHV-4, fibrocytes became infected, indicating that the unidentified 50% of the infected cells are most probably fibrocytes. In this study, in vivo-related in vitro genital tract models were successfully established and the early stage of the pathogenesis of a genital infection was clarified: BoHV-4 starts with a productive infection of epithelial cells in the reproductive tract, forming small foci followed by a non-productive infection of surveilling monocytic cells which help BoHV-4 to invade into deeper tissues.

Us3 and Us9 proteins contribute to the stromal invasion of bovine herpesvirus 1 in the respiratory mucosa.

Bovine herpesvirus 1 (BHV-1) infection may lead to conjunctivitis, upper respiratory tract problems, pneumonia, genital disorders and abortion. BHV-1 is able to spread quickly in a plaque-wise manner and invade by breaching the basement membrane (BM) barrier in the respiratory mucosa. BHV-1 Us3, a serine/threonine kinase, induces a dramatic cytoskeletal reorganization and BHV-1 Us9, a tail-anchored membrane protein, is required for axonal transport of viruses in neurons. In this study, we investigated the role of Us3 and Us9 during BHV-1 infection in the respiratory mucosa. First, we constructed and characterized BHV-1 Us3 null, Us9 null and revertant viruses. Then, we analysed the viral replication and plaque size (latitude) in Madin-Darby bovine kidney (MDBK) cells and the respiratory mucosa as well as viral penetration depth underneath the BM of the respiratory mucosa when inoculated with these recombinant viruses. Knockout of Us3 resulted in a 1 log10 reduction in viral titre and plaque size (latitude) in MDBK cells and the trachea mucosa. There were no defects in the cell-to-cell spread observed for BHV-1 Us9 null virus. Both BHV-1 Us3 null and Us9 null viruses showed a significant reduction of plaque penetration underneath the BM; however, penetration was not completely inhibited. In conclusion, the current findings demonstrated that Us3 and Us9 play an important role in the invasion of BHV-1 through the BM of the respiratory mucosa, which shows the way forward for research-based attenuation of viruses in order to make safer and better-performing vaccines.

Effects of biochars derived from chicken manure and rape straw on speciation and phytoavailability of Cd to maize in artificially contaminated loess soil.

While biochar can reduce the bioavailability of heavy metals in acidic soils and reduce their risk of entering the food chain, conditions for alkaline soils such as loess soils with high pH values, high carbonate content and low organic matter content remain unclear. Pot experiments were conducted to assess the effects of four rates (1%, 5%, 10%, and 15% w/w) of biochars prepared at 600 °C from chicken manure and rape straw (CBC and RBC) on soil properties, Cd speciation and phytoavailability, and plant growth in Cd contaminated (20 mg kg-1) light sierozem using maize (Zea mays L.) as an indicator plant. Biochar additions significantly (P < 0.05) increased soil pH values, cation exchange capacity (CEC) and soil organic matter (OM). The results showed that Cd speciation turned somewhat into stable state as biochar application increased. When CBC and RBC was applied at the rate of 15%, the content of acid-extractable Cd decreased only by 16.3% and 11.64%, respectively. The uptake of Cd by maize shoots scarcely decreased with CBC and RBC amendment at the rate of 1% and 5%, respectively. Although it seemed that additions of more than 5% CBC or RBC significantly (P < 0.05) reduced Cd contents in maize shoots, maize growth was largely inhibited due to the high value of soil pH. These results could provide different implications for immobilization remediation of loess soils (e.g., light sierozem) contaminated with Cd.

Productive replication of nephropathogenic infectious bronchitis virus in peripheral blood monocytic cells, a strategy for viral dissemination and kidney infection in chickens.

In the present study, the replication kinetics of nephropathogenic (B1648) and respiratory (Massachusetts-M41) IBV strains were compared in vitro in respiratory mucosa explants and blood monocytes (KUL01(+) cells), and in vivo in chickens to understand why some IBV strains have a kidney tropism. B1648 was replicating somewhat better than M41 in the epithelium of the respiratory mucosa explants and used more KUL01(+) cells to penetrate the deeper layers of the respiratory tract. B1648 was productively replicating in KUL01(+) monocytic cells in contrast with M41. In B1648 inoculated animals, 10(2.7-6.8) viral RNA copies/100 mg were detected in tracheal secretions at 2, 4, 6, 8, 10 and 12 days post inoculation (dpi), 10(2.4-4.5) viral RNA copies/mL in plasma at 2, 4, 6, 8, 10 and 12 dpi and 10(1.8-4.4) viral RNA copies/10(6) mononuclear cells in blood at 2, 4, 6 and 8 dpi. In M41 inoculated animals, 10(2.6-7.0) viral RNA copies/100 mg were detected in tracheal secretions at 2, 4, 6, 8 and 10 dpi, but viral RNA was not demonstrated in plasma and mononuclear cells (except in one chicken at 6 dpi). Infectious virus was detected only in plasma and mononuclear cells of the B1648 group. At euthanasia (12 dpi), viral RNA and antigen positive cells were detected in lungs, liver, spleen and kidneys of only the B1648 group and in tracheas of both the B1648 and M41 group. In conclusion, only B1648 can easily disseminate to internal organs via a cell-free and -associated viremia with KUL01(+) cells as important carrier cells.

Early events of canine herpesvirus 1 infections in canine respiratory and genital mucosae by the use of ex vivo models.

Canine herpesvirus 1 (CaHV-1) causes a systemic disease in newborn puppies, kennel cough at all ages and genital lesions in adult dogs. The aim of the present study was to elucidate the viral behavior during the early stage of infection in respiratory and genital mucosae, the portals of entry for CaHV-1 by the use of ex vivo explants. CaHV-1 infected and replicated in respiratory and vaginal mucosae in a plaque wise manner. CaHV-1 started to penetrate the basement membrane (BM) only after 48 h post inoculation (hpi) in respiratory mucosal explants, but already after 24 hpi in vaginal explants. The plaque latitude and penetration depth increased over time and both were larger in the vaginal explants compared to the respiratory mucosal explants. The canine respiratory and genital mucosal explants were suitable to study the early pathogenesis of CaHV-1. CaHV-1 showed a better capacity to replicate and invade vaginal mucosa compared to respiratory mucosa, based on the latitude and penetration depth of the plaques of viral antigen positive cells.

Immunity raised by recent European subtype 1 PRRSV strains allows better replication of East European subtype 3 PRRSV strain Lena than that raised by an older strain.

Stable spatial distribution of porcine reproductive and respiratory syndrome (PRRSV)-1 subtypes in Europe is accompanied by a strong population immunity induced by local PRRSV strains. In the present study, it was examined if the immunity induced by three West European subtype 1 PRRSV strains (2007 isolate 07V063 and 2013 isolates 13V091 and 13V117) offers protection against the highly virulent East European subtype 3 PRRSV strain Lena. The number of fever days was greater (p < 0.05) in the control group (7.6 ± 1.7 days) compared to the immune groups (07V063-immune: 4.0 ± 1.2 days, 13V091-immune: 4.6 ± 1.1 days, 13V117-immune: 4.0 ± 2.9 days). In all groups, protection was characterized by reduction (p < 0.05) of AUC values of nasal shedding (control: 14.6, 07V063-immune: 3.4, 13V091-immune: 8.9, 13V117-immune: 8.0) and viremia (control: 28.1, 07V063-immune: 5.4, 13V091-immune: 9.0, 13V117-immune: 8.3). Reduction of respiratory disease, nasal shedding (mean AUC and mean peak values) and viremia (mean AUC and mean peak values) was more pronounced in 07V063-immune (p < 0.05) than in 13V091-immune and 13V117-immune animals. Inoculation with subtype 1 PRRSV strains caused priming of the Lena-specific virus neutralization antibody response. Upon challenge with Lena, we observed a very strong serological booster effect for neutralizing antibodies against strains used for the first inoculation. Our results indicate that inoculation with subtype 1 PRRSV strains can partially protect against antigenically divergent subtype 3 strains. The lower protection level elicited by recently isolated subtype 1 PRRSV strains may impair the outcome of the spatial expansion of subtype 3 strains from East Europe to West Europe.

Replication characteristics of equine herpesvirus 1 and equine herpesvirus 3: comparative analysis using ex vivo tissue cultures.

Replication kinetics and invasion characteristics of equine herpesvirus-1 and -3 (EHV-1/-3) in nasal and vaginal mucosae were compared using explants. The explants were cultured during 96 h with little change in viability. The tissues were inoculated with EHV-1 03P37 (neuropathogenic), 97P70 (abortigenic) and EHV-3 04P57, collected at 0, 24, 48 and 72 h post inoculation (pi) and stained for viral antigens. Both EHV-1 and EHV-3 replicated in a plaquewise manner. The plaques were already observed at 24 h pi, their size increased over time and did not directly cross the basement membrane (BM). However, EHV-1 infected the monocytic cells (MC) and hijacked these cells to invade the lamina propria. In contrast, EHV-3 replication was fully restricted to epithelial cells; the virus did not breach the BM via a direct cell-to-cell spread nor used infected MC. EHV-1-induced plaques were larger in nasal mucosa compared to vaginal mucosa. The opposite was found for EHV-3-induced plaques. Both EHV-1 strains replicated with comparable kinetics in nasal mucosa. However, the extent of replication of the abortigenic strain in vaginal mucosa was significantly higher than that of the neuropathogenic strain. Two-to-five-fold lower numbers of EHV-1-infected MC underneath the BM were found in vaginal mucosa than in nasal mucosa. Our study has shown that (i) EHV-1 has developed in evolution a predisposition for respiratory mucosa and EHV-3 for vaginal mucosa, (ii) abortigenic EHV-1 replicates better in vaginal mucosa than neuropathogenic EHV-1 and (iii) EHV-3 demonstrated a strict epithelial tropism whereas EHV-1 in addition hijacked MC to invade the lamina propria.

Ex vivo modeling of feline herpesvirus replication in ocular and respiratory mucosae, the primary targets of infection.

Feline herpesvirus 1 (FeHV-1) is a major cause of rhinotracheitis and ocular diseases in cats. In the present study, the viral replication at the primary infection sites was studied using feline respiratory and ocular mucosa explants. The explants of three cats were maintained in an air-liquid culture up to 96 hours without loss of viability. After inoculation with FeHV-1 (C27), no evidence of infection was noted in corneal epithelium, while plaque-wise replication was observed in conjunctival and tracheal mucosae beginning from 24 h post inoculation (hpi). The viral plaque diameters increased over time in trachea and conjunctiva and were larger in tracheal explants than in conjunctival explants at 48 hpi. FeHV-1 penetrated the basement membrane in conjunctival and tracheal explants between 24 and 48 hpi. At 48 and 72 hpi, viral invasion was going deeper in tracheal explants than in conjunctival explants. Our study indicates that FeHV-1 has a better capacity to invade the respiratory mucosa than the conjunctival mucosa, and prefers the conjunctiva, but not the cornea as a portal of entry during ocular infection.

Different clinical, virological, serological and tissue tropism outcomes of two new and one old Belgian type 1 subtype 1 porcine reproductive and respiratory virus (PRRSV) isolates.

In this study, the pathogenic behavior of PRRSV 13V091 and 13V117, isolated in 2013 from two different Belgian farms with enzootic respiratory problems shortly after weaning in the nursery, were compared with the Belgian strain 07V063 isolated in 2007. Full-length genome sequencing was performed to identify their origin. Twelve weeks-old pigs were inoculated intranasally (IN) with 13V091, 13V117 or 07V063 (9 pigs/group). At 10 days post inoculation (dpi), 4 animals from each group were euthanized and tissues were collected for pathology, virological and serological analysis. 13V091 infection resulted in the highest respiratory disease scores and longest period of fever. Gross lung lesions were more pronounced for 13V091 (13%), than for 13V117 (7%) and 07V063 (11%). The nasal shedding and viremia was also most extensive with 13V091. The 13V091 group showed the highest virus replication in conchae, tonsils and retropharyngeal lymph nodes. 13V117 infection resulted in the lowest virus replication in lymphoid tissues. 13V091 showed higher numbers of sialoadhesin(-) infected cells/mm(2) in conchae, tonsils and spleen than 13V117 and 07V063. Neutralizing antibody response with 07V063 was stronger than with 13V091 and 13V117. It can be concluded that (i) 13V091 is a highly pathogenic type 1 subtype 1 PRRSV strain that replicates better than 07V063 and 13V117 and has a strong tropism for sialoadhesin(-) cells and (ii) despite the close genetic relationship between 13V117 and 07V063, 13V117 has an increased nasal replication and shedding, but a decreased replication in lymphoid tissues compared to 07V063.

Replication characteristics of infectious laryngotracheitis virus in the respiratory and conjunctival mucosa.

Avian infectious laryngotracheitis virus (ILTV) is an alphaherpesvirus of poultry that is spread worldwide. ILTV enters its host via the respiratory tract and the eyes. Although ILTV has been known for a long time, the replication characteristics of the virus in the respiratory and conjunctival mucosa are still poorly studied. To study these characteristics, two in vitro explant models were developed. Light microscopy and fluorescent terminal deoxynucleotidyl transferase dUTP nick end-labelling staining were used to evaluate the viability of mucosal explants, which were found to be viable up to the end of the experiment at 96 h of cultivation. The tracheal and conjunctival mucosal explants were inoculated with ILTV and collected at 0, 24, 48 and 72 h post inoculation (p.i.). ILTV spread in a plaque-wise manner in both mucosae. A reproducible quantitative analysis of this mucosal spread was evaluated by measuring plaque numbers, plaque latitude and invasion depth underneath the basement membrane. No major differences in plaque numbers were observed over time. Plaque latitude progressively increased to 70.4 ± 12.9 µm in the trachea and 97.8 ± 9.5 µm in the conjunctiva at 72 h p.i. The virus had difficulty crossing the basement membrane and was first observed only at 48 h p.i. The virus was observed at 72 h p.i. in 56% (trachea) and 74% (conjunctiva) of the plaques. Viability analysis of infected explants indicated that ILTV blocks apoptosis in infected cells of both mucosae but activates apoptosis in bystander cells.