Lesion Margins Research Articles

First Report of Pseudomonas amygdali Causing Leaf Spot on Hibiscus rosa-sinensis in a greenhouse in New York State, U.S.A.

Hibiscus rosa-sinensis, commonly known as the "Chinese hibiscus", is a widely cultivated shrub with ornamental and medicinal applications (Jadhav et al., 2009). However, it is known to be susceptible to a range of pathogens including bacteria (Chase, 1986). In March 2019, Hibiscus rosa-sinensis plants in production at a greenhouse in New York, but originally from Florida, developed widespread, conspicuous leaf spots and associated chlorosis for a month after arrival. "Hibiscus 35-1" was isolated from a suspension of symptomatic plant tissue pieces soaked in sterile deionized water and streaked on potato dextrose agar (PDA) (González-Tobón et al., 2023), resulting in numerous white bacterial colonies with a uniform appearance. Previously we reported the complete species identification for this bacterial isolate using whole genome sequencing via Oxford Nanopore and Illumina platforms (BioProject: PRJNA765326) as a Pseudomonas amygdali with an ANI ≥ 95% ± 0.5%. (González-Tobón et al., 2023). The hypersensitive response (HR) to P. amygdali 35-1 was first evaluated in three tobacco (Nicotiana tabacum) and three tomato (Solanum lycopersicum) plants. An overnight culture was washed, resuspended, adjusted to an OD600 of 0.2-0.3, and used to inoculate two leaves per plant via syringe infiltration (Chakravarthy et al., 2017). HR was observed on all leaves at the inoculation site as early as 48hpi for tobacco and 24hpi for tomato. No symptoms were observed for mock inoculations. The pathogenicity of P. amygdali 35-1 in hibiscus was then tested using six 'Social Butterfly' plants in 4.0 in pots inoculated by hand spraying. Inoculum was made from a 72 h culture on solid King's B medium resuspended in sterile deionized water with 2 drops/L of Tween 80 and sprayed at a concentration of 3.6 x 108 bacteria/mL, as determined by dilution plating. Inoculated plants and six matching controls were sealed inside plastic bags and placed in a randomized complete block in a growth chamber at 30 ºC with a 12h photoperiod for 40 days. No changes were observed on any of the control plants but spotting and chlorosis similar to the original symptoms gradually developed on five of the inoculated plants. After 40 days, seven 2-mm tissue squares were excised from the lesion margins or from the asymptomatic tissue of controls, surface-sterilized with 0.75% NaCl, and plated on King's B. No bacteria were recovered from the control plants but white colonies resembling those of the original inoculation were recovered from several diseased tissue pieces. Genomic DNA was extracted from this bacterium using the New England Biolabs Monarch Genomic DNA Extraction Kit. Polymerase chain reaction (PCR) tests were performed to amplify five genes (16S, cts, gapA, gyrB, and rpoD) typically used for multilocus sequence typing (MLST) (Sarkar & Guttman, 2004). The amplicons were Sanger sequenced and BLAST analyses confirmed 100% match to P. amygdali 35-1 (GenBank: CP084212). To our knowledge, our lab is the first to report P. amygdali causing disease on hibiscus in New York. Similar symptoms on hibiscus plants in Florida were reported in the past as caused by other Pseudomonas species of the P. syringae G group, mainly P. syringae and P. cichorii (Chase, 1986; Gardan et al., 1999: Girard et al., 2021; Jones et al., 1986). Given the lack of consistent reporting on bacterial leaf spot pathogens of hibiscus, as well as the economic importance of this ornamental, further research on the disease is needed.

The Role and Potential of Digital Breast Tomosynthesis in Neoadjuvant Systemic Therapy Evaluation for Optimising Breast Cancer Management: A Pictorial Essay.

Neoadjuvant Therapy (NT) has become the gold standard for treating locally advanced Breast Cancer (BC). The assessment of pathological response (pR) post-NT plays a crucial role in predicting long-term survival, with Contrast-Enhanced Magnetic Resonance Imaging (MRI) currently recognised as the preferred imaging modality for its evaluation. Traditional imaging techniques, such as Digital Mammography (DM) and Ultrasonography (US), encounter difficulties in post-NT assessments due to breast density, lesion changes, fibrosis, and molecular patterns. Digital Breast Tomosynthesis (DBT) offers solutions to prevalent challenges in DM, such as tissue overlap, and facilitates a comprehensive assessment of lesion morphology, dimensions, and margins. Studies suggest that DBT correlates more accurately with pathology than DM and US, showcasing its potential advantages. This pictorial essay demonstrates the potential of DBT as a complementary tool to DM for assessing pR after NT, including instances of true and false-positive assessments correlated with histopathological findings. In conclusion, DBT emerges as a valuable adjunct to DM, effectively addressing its limitations in post-NT assessment. The technology's potential to diminish tissue overlap, improve discrimination, and provide multi-dimensional perspectives demonstrates promising results, indicating its utility in scenarios where MRI is contraindicated or inaccessible.

First Report of Exserohilum rostratum Causing Leaf Blight on Calathea orbifolia in China.

Peacock plant (Calathea orbifolia (Linden) H.A.Kenn.) is cultivated as a valuable houseplant in China. In September 2024, leaf blight was found on this plant in a field located in Shenzhen (22°35'55"N, 113°59'21"E), Guangdong Province, China. The area investigated was ~900 m2. The disease incidence was ~70% of 100 plants. Initial symptoms of brown spots appeared on leaves, gradually becoming enlarged, elongated, 0.5 to 10 × 0.2 to 4.5 cm, irregular, dark brown, with a yellow halo, and leaves blighted and desiccated. Ten symptomatic leaves were collected for pathogen isolation. Leaf pieces (5×5 mm2) of lesion margins were surface disinfected with 1% NaClO for 1 min, 70% ethanol for 30 s, rinsed in sterile distilled water, placed on PDA, and incubated at 25°C. After 3 days, fungal colonies were purified by a single hyphal tip method on PDA. Three isolates (MBSZU 24-001 to MBSZU 24-003) with similar morphology were obtained. Colonies on PDA were 48 to 54 mm in diameter after 1 week at 25°C, then gray, flat with entire edges, aerial mycelia dense, and dark in the center and gray at the margin on the reverse side after 2 weeks. Conidiophores were olivaceous-brown, cylindrical, septate, unbranched, up to 230 × 3.4 to 5 µm. Conidiogenous cells were integrated, straight to curved, proliferating sympodial, 25 to 60 × 4.5 to 7 µm. Conidia were straight to slightly curved, ellipsoid, pale olivaceous brown to dark olivaceous brown, 2-6-distoseptate, with a distinct basal hilum, 26.9 to 70 × 11.4 to 21.6 µm (n = 50). Morphological characteristics of all isolates resembled the Exserohilum (Hernández-Restrepo et al. 2018). The internal transcribed spacer (ITS) region, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation elongation factor 1-alpha (tef1-α) genes were amplified using primer pairs ITS5/ITS4 (White et al. 1990), GPD1/GPD2 (Berbee et al. 1999), and EF1-983F/EF1-2218R (Rehner and Buckley 2005), respectively. Sequences were deposited in GenBank (ITS: PQ498876 to PQ498878; GAPDH: PQ509760 to PQ509762; tef1-α: PQ509763 to PQ509765). In the BLAST analysis, the ITS, GAPDH, and tef1-α sequences showed 99.50% (599 bp out of 602 bp), 98.89% (536 bp out of 542 bp), and 99.78% (926 bp out of 928 bp), respectively, to E. rostratum (CBS 325.87). Maximum likelihood and Bayesian inference phylogenetic analyses of the concatenated three genes identified all isolates as E. rostratum. To test pathogenicity, both wounded and unwounded healthy leaves were used in this experiment. All leaves were wiped with 0.1% NaClO and then rinsed three times with sterile water. Conidia suspensions (15 µl of 1 × 106 conidia/ml) of each isolate grown on PDA at 25°C for 3 weeks were placed on all samples using the attached leaf assay. Control leaves were mock-inoculated with sterile distilled water. Ten replications were conducted for each treatment and repeated twice. Plants were incubated at 25°C with 80 to 85% relative humidity. After 10 days, all inoculated leaves had brown lesions with yellow halos, while control leaves remained asymptomatic. E. rostratum was consistently reisolated from the lesions on PDA to complete Koch's postulates. Prior to this study, E. rostratum was identified as a causal agent of leaf blight on Calathea picturata in China (Taiwan, Chern et al. 2011). This is the first report of leaf blight caused by E. rostratum on C. orbifolia in China and worldwide. These findings will provide insights for epidemiological investigations and future approaches to managing this disease.

Feasibility/clinical utility of half-Fourier single-shot turbo spin echo imaging combined with deep learning reconstruction in gynecologic magnetic resonance imaging.

When antispasmodics are unavailable, the periodically rotated overlapping parallel lines with enhanced reconstruction (PROPELLER; called BLADE by Siemens Healthineers) or half Fourier single-shot turbo spin echo (HASTE) is clinically used in gynecologic MRI. However, their imaging qualities are limited compared to Turbo Spin Echo (TSE) with antispasmodics. Even with antispasmodics, TSE can be artifact-affected, necessitating a rapid backup sequence. This study aimed to investigate the utility of HASTE with deep learning reconstruction and variable flip angle evolution (iHASTE) compared to conventional sequences with and without antispasmodics. This retrospective study included MRI scans without antispasmodics for 79 patients who underwent iHASTE, HASTE, and BLADE and MRI scans with antispasmodics for 79 case-control matched patients who underwent TSE. Three radiologists qualitatively evaluated image quality, robustness to artifacts, tissue contrast, and uterine lesion margins. Tissue contrast was also quantitatively evaluated. Quantitative evaluations revealed that iHASTE exhibited significantly superior tissue contrast to HASTE and BLADE. Qualitative evaluations indicated that iHASTE outperformed HASTE in overall quality. Two of three radiologists judged iHASTE to be significantly superior to BLADE, while two of three judged TSE to be significantly superior to iHASTE. iHASTE demonstrated greater robustness to artifacts than both BLADE and TSE. Lesion margins in iHASTE had lower scores than BLADE and TSE. iHASTE is a viable clinical option in patients undergoing gynecologic MRI with anti-spasmodics. iHASTE may also be considered as a useful add-on sequence in patients undergoing MRI with antispasmodics.

Development of Deep Learning-Based Virtual Lugol Chromoendoscopy for Superficial Esophageal Squamous Cell Carcinoma.

Lugol chromoendoscopy has been shown to increase the sensitivity of detection of esophageal squamous cell carcinoma (ESCC). We aimed to develop a deep learning-based virtual lugol chromoendoscopy (V-LCE) method. We developed still V-LCE images for superficial ESCC using a cycle-consistent generative adversarial network (CycleGAN). Six endoscopists graded the detection and margins of ESCCs using white-light endoscopy (WLE), real lugol chromoendoscopy (R-LCE), and V-LCE on a five-point scale ranging from 1 (poor) to 5 (excellent). We also calculated and compared the color differences between cancerous and non-cancerous areas using WLE, R-LCE, and V-LCE. Scores for the detection and margins were significantly higher with R-LCE than V-LCE (detection, 4.7 vs. 3.8, respectively; p < 0.001; margins, 4.3 vs. 3.0, respectively; p < 0.001). There were nonsignificant trends towards higher scores with V-LCE than WLE (detection, 3.8 vs. 3.3, respectively; p = 0.089; margins, 3.0 vs. 2.7, respectively; p = 0.130). Color differences were significantly greater with V-LCE than WLE (p < 0.001) and with R-LCE than V-LCE (p < 0.001) (39.6 with R-LCE, 29.6 with V-LCE, and 18.3 with WLE). Our V-LCE has a middle performance between R-LCE and WLE in terms of lesion detection, margin, and color difference. It suggests that V-LCE potentially improves the endoscopic diagnosis of superficial ESCC.

Chondrosarcoma of the Flat Bones: Differential Survival Between High-Grade Lesions of the Pelvis and Scapula.

Flat bone chondrosarcomas have worse outcomes than extremity tumors, but there is no data directly comparing the different flat bones. The aim of this study was to examine differences in recurrence and survival between pelvic and scapular chondrosarcoma. One hundred and sixty-nine (42 scapula, 127 pelvic) patients with chondrosarcoma who underwent surgical resection were reviewed. High-grade tumors were defined as lesions that were Grade 3 or dedifferentiated on surgical pathology. Patients with low-intermediate grade lesions of the scapula were more likely to have positive margins during definitive surgical management (14% vs. 3%, OR 5, 95% CI [1.15, 22.6], p = 0.02), however, this did not translate to differences in recurrence or survival (p > 0.05). The presence of an associated soft tissue mass in low-intermediate grade lesions was not associated with increased recurrence. Among high-grade tumors, patients with scapular lesions had worse DSS (HR 2.99, 95% CI [1.05, 8.51], p = 0.04). One- and 2-year DSS for high-grade pelvic tumors was 75% and 57%, respectively, and 50% and 33% for scapular tumors. Survival for high-grade chondrosarcoma of the flat bones is poor, particularly for those of the scapula. Despite a higher rate of positive margins for low-intermediate grade lesions of the scapula, there was no significant difference in survival compared to low-intermediate grade lesions of the pelvis. Additional studies with larger sample sizes are needed to further elucidate differences between these locations.

Diagnostic Confidence of Contrast-Enhanced T1-Weighted MRI for the Detection of Brain Metastases: 3D FSE-vs. 3D GRE-Based Sequences.

This retrospective study evaluated the utility of contrast-enhanced (CE) T1-weighted 3D fast spinecho-based SPACE sequences for brain metastasis detection on 3T MRI compared to gradient recalled-echo-based 3D fast low-angle shot (FLASH) sequence. We identified all patients at a single institution who underwent SPACE and 3D FLASH sequences as part of a practice quality improvement project. Their medical records were retrospectively reviewed. Five certified neuroradiologists reviewed the images, with at least 2 weeks separation between scoring sequences for the same patient. The following parameters were evaluated: number of metastatic lesions, number of indeterminate lesions, lesion margin, contrast-to-noise ratio (CNR), extent of image artifacts, and overall image quality. CNR was also quantified for solidly enhancing lesions > 1 cm. We identified 220 patients who underwent SPACE and 3D FLASH sequences (the order of the sequences was equally distributed). Of these, 79 had brain metastases on imaging, and 7 were excluded; thus, 72 patients were included in the study. Twenty patients were scored by 2 radiologists. Out of the 92 evaluations, SPACE detected more lesions than did 3D FLASH in 35, while 3D FLASH detected more lesions in 10. More indeterminate lesions were seen on 3D FLASH (27) than on SPACE (9). For lesion margin, CNR, and overall image quality on a Likert scale, SPACE performed significantly better than did 3D FLASH, with less image artifacts (P < 0.00001). Higher quantitative CNRs were found on SPACE than on 3D FLASH images, although this result was not statistically significant (median = 22.9 vs. 15.5, respectively, P = 0.134). There was a high inter-reader lesion detection concordance with Krippendorf's alpha ordinals at 0.962 for SPACE, 0.870 for 3D FLASH, and 0.918 for the two sequences combined. Compared with 3D FLASH, the SPACE sequence detected more metastatic lesions and was rated higher for image quality, lesion margin, and CNR, with fewer artifacts. Importantly, the SPACE sequence resulted in increased reader confidence, with fewer indeterminate lesions detected. FLASH = fast low-angle shot; FSE = fast spin-echo; GRE = gradient-recalled echo; MP-RAGE = magnetization-prepared rapid gradient echo; SPACE = Sampling Perfection with Application-optimized Contrasts using different flip angle Evolution; VIBE = volumetric interpolated breath-hold examination.

Magnetic Resonance Imaging Assessment of Morphological Changes and Molecular Behavior to Evaluate Treatment Response of Brain Metastatic Lesions After Stereotactic Radiosurgery.

Brain metastases (BMs) are the most common type of intracranial tumors, frequently arising from primary cancers such as lung, breast, melanoma, and renal cell carcinoma. Magnetic resonance imaging (MRI) plays a crucial role in assessing both the morphological and molecular characteristics of BMs, particularly in evaluating treatment response following radiosurgery. However, the interpretation of these imaging changes remains complex, often influencing clinical decision-making. This study aims to evaluate the morphological changes and molecular behavior of BMs postradiosurgery using MRI to assess treatment response. A retrospective review was conducted at a high specialty medical center, including 41 patients with BMs treated with stereotactic radiosurgery (SRS) from 2018 to 2022. Patients had a baseline MRI (pre-SRS) prior to treatment and follow-ups at 2-3 months (MRI-2) and 5-6 months (MRI-3). The response assessment in neuro-oncology brain metastases (RANO-BM)criteria were used, and T1/T2 matching was analyzed for each follow-up. Logistic regression was performed relating the T1/T2 matching and susceptibility areas (susceptibility-weighted imaging (SWI)) for MRI-2 and MRI-3. Cross tables were created regarding treatment response and demographic characteristics according to Pearson's Chi-squared test. The mean age was 56.7 years; 53.7% (n= 22) were female. Primary tumors included lung (29.3%, n= 12), breast (19.5%, n= 8), colon (12.2%, n= 5), and melanoma and kidney tumors (7.3%, n= 3). Post-SRS changes included transitions from solid to cystic lesions, reduced perilesional edema, size reduction, and increased areas of magnetic susceptibility. A mixed pattern (areas of T1/T2 match + mismatch) was noted at lesion margins during follow-ups (MRI-2: 70.7% (n= 29), MRI-3: 68.3% (n= 28)). Most patients exhibited a partial response at MRI-2 (43.9%, n= 18), while at MRI-3, disease progression occurred (43.9%, n= 18) due to an increase in lesion number. Logistic regression linking T1/T2 matching and SWI demonstrated a significantly central-peripheral SWI distribution for T1/T2 match during both follow-ups (MRI-2:p = 0.005, R2: 0.52; MRI-3:p = 0.002, R2: 0.56). SWI distribution was higher when a mixed T1/T2 matching was present. Significant associations were found with systemic treatment and response type at MRI-2 (p =0.001), predominantly showing a partial response for those receiving chemotherapy + targeted therapy. SWI and T1/T2 mismatch are valuable tools reflecting changes in the tumor microenvironment postradiosurgery, aiding in treatment response monitoring. The appearance of susceptibility areas may precede changes in the enhancement of the lesion margin. Short-term follow-ups (2-3 months) are crucial due to prevalent progression, marked primarily by the appearance of new lesions in approximately 50% of patients.

First Report of Lelliottia amnigena Causing Soft Rot on Purple Stem Mustards in China.

The purple stem mustards (Brassica rapa subsp. chinensis var. purpuraria) (Govaerts R, 2003) are widely cultivated along Yangtze River Valley in China, which is famous for its flavorful and nutritious edible stalks (Wang et al., 2022). In February 2023, a disease of soft rot was observed in multiple purple stem mustards planting fields in Wuhan city, Hubei province, China (30.41°N, 114.22°E). Disease incidence rates were almost 20 to 30% in the planting area (5 ha in size), causing severe economic loss. Infected plants displayed water-soaked symptoms at the base of the petioles, emitting a foul soft rot odor. The severely infected petioles, stems and roots exhibit pus symptoms leading to plant death. To identify the causal agent, small pieces of soft rot symptomatic tissue were cut from the margin of necrotic lesions and surface disinfected with 75% (v/v) ethanol for 30 seconds, followed by three successive rinses with sterile distilled water. The exudates from the clipped tissues were serially diluted and then incubated onto nutrient agar (NA) plates to obtain purified strains at 28°C for 48 hours (Koike et al., 2002). After incubation, 15 strains were obtained and the colonies of all strains were Gram-negative, aerobic, small, round, convex, whitish to dull white, and had smooth slimy edges. Three single bacterial strains CT020801 - CT020803, which were individually isolated from three different diseased samples, were selected as representative strains for further study. Biochemical tests using the BIOLOG GENIII microplate system (Biolog, Hayward, CA, USA) revealed that these strains were positive for methyl red, pectin, dextrin, D-Cellobiose, β-galactosidase, citrate, and maltose, but negative for indole, arginine dihydrolase, urease, ornithine decarboxylase, and gelatinase tests. The 16S ribosomal RNA gene and the three housekeeping genes, atpD, rpoB, and recN were amplified using genomic DNA of Lelliottia amnigena NCTC12124T as the template, with specially designed primers. All amplified fragments were sequenced and deposited in GenBank with accession numbers OQ954706-OQ954707, OQ954713, and OQ953873-OQ953881. BLAST alignments of the 16S rRNA, atpD, rpoB and recN sequences revealed that the sequences of Strain CT020801-03 exhibited the highest identity (100%, ≥97.97%, ≥98.85% and ≥94.52%, respectively) with L. amnigena （Figure S2）. Phylogenetic tree analysis based on multilocus sequence joint 16S rRNA - atpD - rpoB - recN revealed that CT020801 - CT020803 and L. amnigena clustered together in the same clade (Carrie et al., 2013). These results were consistent with those reported for Lelliottia amnigena (Osei et al., 2022). To confirm pathogenicity, healthy base petioles of three-week-old purple stem mustards seedlings were stab inoculated with 20 μL bacterial suspensions (approximately 108 CFU/mL) and then incubated at 28°C and 80% relative humidity in a growth chamber. A sterile liquid NB medium served as the negative control. The test was repeated thrice with each test consisting of five seedlings per treatment. After three days, soft rot symptoms appeared on the stem bases of the inoculated plants, consistent with the initial symptoms observed in the field. Control plants showed no symptoms. The strains were successfully re-isolated from symptomatic plants and identified as L. amnigena, fulfilling Koch's postulates. L. amnigena, a member of the Enterobacter genus (Birlutiu et al., 2023; Brady et al., 2013; Izard et al., 1981), is commonly found in soil and has been identified as an opportunistic pathogen responsible for plant soft rot disease (Reyes-García et al., 2020; Schroeder et al., 2009; Wu et al., 2023). Previous studies have linked L. amnigena to soft rot disease in potato tubers in China. However, this study marks the first documented case of L. amnigena causing soft rot disease in purple stem mustards in China. This discovery expands the known host range of the pathogenic bacteria and will help to provide essential information for making effective measures to manage this disease.

Fluoroscopy-Guided Percutaneous Transthoracic Needle Lung Biopsy with the Aid of Planning Cone-Beam CT: Diagnostic Accuracy and Complications.

Background: Fluoroscopy-guided PTNB for fluoroscopy-identifiable lung lesions has been suggested as a useful method for the pathological diagnosis of lung lesions; however, it is lacking in accuracy and safety compared to CT-guided PTNB. Thus, we aimed to investigate the diagnostic accuracy and complications of fluoroscopy-guided percutaneous transthoracic needle biopsy (PTNB) with the aid of pre-procedural planning cone-beam computed tomography (CBCT) in order to take advantage of their respective strengths. Methods: A total of 255 fluoroscopy-guided PTNBs with the aid of planning CBCT were performed. Pre-procedural planning CBCT was conducted to calculate the shortest length from the skin puncture site to the margin of the target lesion for the needle trajectory. No intra-procedural CBCT was performed. The diagnostic performance of fluoroscopy-guided PTNB with the aid of planning CBCT was calculated. The prognostic factors for diagnostic failures and complications were evaluated using logistic regression analysis. Results: The accuracy, sensitivity, specificity, PPV, and NPV were 97.3%, 88.0%, 90.9%, 100%, and 62.5%, respectively. There were 29 diagnostic failures (11.8%), and the multivariable analysis showed that a longer lesion depth on CBCT and a shorter specimen length were each associated with diagnostic failure (p = 0.010 and 0.012, respectively). Complications occurred in 34 PTNBs (13.3%). The multivariable analysis showed that an increased total number of biopsies per lesion, a longer length of lung aeration via needle insertion, a smaller lesion size on CT imaging (≤20 mm), and the presence of an air bronchogram were associated with the occurrence of complications (p = 0.027, <0.001, 0.003, and 0.020, respectively). Conclusions: Excellent diagnostic accuracy was obtained by fluoroscopy-guided PTNB with the aid of planning CBCT. Compared to that of CT- or CBCT-guided PTNB, the procedure-related complication rate was acceptably low, but the radiation dose to patients could be potentially reduced.

A Multi-label Artificial Intelligence Approach for Improving Breast Cancer Detection With Mammographic Image Analysis.

Breast cancer remains a major global health concern. This study aimed to develop a deep-learning-based artificial intelligence (AI) model that predicts the malignancy of mammographic lesions and reduces unnecessary biopsies in patients with breast cancer. In this retrospective study, we used deep-learning-based AI to predict whether lesions in mammographic images are malignant. The AI model learned the malignancy as well as margins and shapes of mass lesions through multi-label training, similar to the diagnostic process of a radiologist. We used the Curated Breast Imaging Subset of Digital Database for Screening Mammography. This dataset includes annotations for mass lesions, and we developed an algorithm to determine the exact location of the lesions for accurate classification. A multi-label classification approach enabled the model to recognize malignancy and lesion attributes. Our multi-label classification model, trained on both lesion shape and margin, demonstrated superior performance compared with models trained solely on malignancy. Gradient-weighted class activation mapping analysis revealed that by considering the margin and shape, the model assigned higher importance to border areas and analyzed pixels more uniformly when classifying malignant lesions. This approach improved diagnostic accuracy, particularly in challenging cases, such as American College of Radiology Breast Imaging-Reporting and Data System categories 3 and 4, where the breast density exceeded 50%. This study highlights the potential of AI in improving the diagnosis of breast cancer. By integrating advanced techniques and modern neural network designs, we developed an AI model with enhanced accuracy for mammographic image analysis.

First Report of Alternaria alternata Causing Leaf Spot of Magnolia delavayi in China

Magnolia delavayi, known as Delavay's magnolia or Chinese evergreen magnolia, is an enchanting deciduous tree native to southwestern China (Xu et al. 2020). In March 2024, leaf spots were observed on M. delavayi plants at Qujing Normal University (25.527°N, 103.744°E), Qujing, Yunnan, China. About 80% of the trees (n=92) on campus were infected, with infected leaves accounting for about 10 to 30% of all leaves. The leaf spots exhibited a round or oval shape, with a yellowish-brown center and a brown outer ring. As the spots expanded, the leaves gradually dried up and fell off. Five symptomatic leaves were collected from the middle sections of five diseased plants. Tissue samples (5 × 5 mm) were excised from the lesion margins and subjected to surface sterilization in 75% ethanol for 30 seconds and 1% sodium hypochlorite for 60 seconds. They were then rinsed three times with sterile water and dried on sterilized paper. The samples were plated on potato dextrose agar (PDA) and incubated at 28 °C for 3 days. Developing colonies were retransferred to new PDA and purified by the hyphal tip technique (Senanayake et al. 2020). Two representative isolates (SYL1 and SYL2) were selected and confirmed to be the same species based on morphological characteristics and molecular identification. On PDA, the fungal colonies initially appeared grayish-white and turned dark gray over time with cotton-like aerial hyphae on the surface. The conidia are light brown, pear-shaped or rod-shaped, with 1 to 4 transverse septa and 0 to 1 oblique septa, and measured 5.4 to 12.7 × 10.4 to 25.7 μm (n = 50). Molecular identification was performed by partial sequencing of the internal transcribed spacer (ITS: ITS4/ITS5) region, glyceraldehyde 3-phosphate dehydrogenase (GAPDH:gpd1/gpd2), translation elongation factor 1-alpha (TEF1:EF1/EF2), and Alternaria major allergen (Alt a 1:Alt-for/Alt-rev) (Woudenberg et al. 2015). The resulting sequences of ITS (PP951882, PP951883), GAPDH (PP968942, PP968943), Alt a 1(PP968940, PP968941) and TEF1 (PP968938, PP968939) were deposited in GenBank. BLAST analyses revealed that the sequences from these two isolates showed 100% identity with those from Alternaria alternata strains for each gene. Additionally, a multigene phylogenetic analysis revealed a distinct group containing SYL1, SYL2 and other A. alternata strains with a posterior probability of 100%. The morphological and phylogenetic evidence collectively suggested that the two isolates belonged to A. alternata. Strain SYL1 was used for the pathogenicity test on three 5-year-old M. delavayi plants on campus. Three healthy leaves of each plant were punctured with a sterile needle and sprayed with 20 μl of spore suspension (1 × 106 conidia/ml). Similarly, another nine wounded leaves were sprayed with 20 μl of sterile water. All inoculated leaves were enclosed in plastic bags for 48 hours. Disease symptoms were observed on leaves inoculated with spore suspension after 7 days, while control plants remained healthy. This experiment was repeated three times with same results. To fulfill Koch’s postulates, A. alternata was reisolated from the inoculated leaves and identified based on morphological characteristics and ITS sequencing. To our knowledge, this is the first report of A. alternata causing leaf spot on M. delavayi in China. This research expands the known host range of this pathogen and provides relevant information to the design of disease management strategies.

Contrast-Enhanced Digital Mammography for the Diagnosis and Determination of Extent of Disease in Invasive Lobular Carcinoma: Our Experience and Literature Review.

Contrast-enhanced imaging, including magnetic resonance imaging and, more recently, contrast-enhanced digital mammography (CEM), is indicated for the precise diagnosis of invasive lobular carcinoma (ILC). The aim of our study was to further validate the use of CEM for evaluation of extent of disease in ILC cases, with digital breast tomosynthesis (DBT) as an adjunct. A retrospective, institutional review board approved study was conducted in a tertiary medical center. All CEM examinations performed on ILC patients between 2017 and 2023 were reread by 2 dedicated breast radiologists. Clinical data and pathology reports were retrieved from electronic medical records. The longest diameter of the enhancing lesion was correlated to pathology findings. In addition, for each case, the readers provided brief commentary on the added value of DBT. Twenty-four CEM examinations were evaluated. The subjects in the study cohort were on average older than expected for ILC patients (74 vs 63 years) and were unable to undergo breast magnetic resonance imaging due to advanced age and comorbidities. Three subjects were treated with neoadjuvant therapy and thus were excluded from the correlation to pathology analysis. Enhancing lesions, ranging from 4-75 mm, strongly correlated to pathology results, with statistical significance. This was demonstrated for mass and nonmass lesions (r = 0.94, P < 0.001 and r = 0.99, P = 0.002, respectively). For most lesions (17/24, 71%), readers remarked that the addition of DBT allowed for improved characterization of lesion margins, mainly detecting architectural distortion. When compared with the pathology findings, ILC was accurately diagnosed and assessed using CEM. The addition of DBT was reported by the interpreting radiologists as a valuable adjunct for margin analysis.

Navigation of video-assisted thoracoscopic surgery using electromagnetic versus CT-guided localization (NOVEL): a study protocol for comparing procedural success and complication rates in a prospective, multicenter, randomized controlled, non-inferiority phase III trial.

The rise of low-dose computed tomography (LDCT) has increased the detection of small pulmonary nodules, demanding more effective localization techniques for their resection. Minimally invasive resection utilizing video-assisted thoracoscopic surgery (VATS) is a critical method for treating these nodules. However, traditional computed tomography (CT)-guided localization has limitations such as invasiveness and patient discomfort. The current gap in knowledge relates to the potential advantages of electromagnetic navigation bronchoscopy (ENB) in reducing complications and improving procedural efficiency. The NOVEL trial evaluates the non-inferiority of ENB-guided labeling against CT-guided puncture for lung nodule localization. This multicenter, randomized, controlled, non-inferiority phase III trial includes 156 participants across four Chinese hospitals, randomized to undergo either ENB-guided or CT-guided localization prior to VATS sub-lobar resection. Randomization is performed using sealed opaque envelopes to ensure allocation concealment. Primary outcomes are the procedural success rates and complication rates of both techniques, with secondary outcomes including procedure times and lesion margins. The NOVEL trial aims to provide a detailed comparison of ENB-guided versus CT-guided localization for small pulmonary nodules. Establishing the safety and efficacy of the ENB method could significantly influence clinical practices and improve patient outcomes. This trial was registered with the Medical Research Registration Platform (https://www.medicalresearch.org.cn), registration number MR-31-24-018575.

Implantable Microneedle-Mediated Eradication of Postoperative Tumor Foci Mitigates Glioblastoma Relapse.

Glioblastoma multiforme (GBM) remains incurable despite multimodal treatments after surgical debulking. Almost all patients with GBM relapse within a narrow margin (2-3cm) of the initial resected lesion due to the unreachable residual cancerous cells. Here, a completely biodegradable microneedle for surgical cavity delivery glioblastoma-associated macrophages (GAMs)-activating immune nano-stimulator that mitigates glioblastoma relapse is reported. The residual tumor lesion-directed biocompatible microneedle releases the nano-stimulator and toll-like receptor 9 agonist in a controlled manner until the microneedles completely degrade over 1 week, efferently induce in situ phonotypic shifting of GAMs from anti- to pro-inflammatory and the tumor recurrence is obviously inhibited. The implantable microneedles offer a significant improvement over conventional transdermal ones, as they are 100% degradable, ensuring safe application within surgical cavities. It is also revealed that the T cells are recruited to the tumor niche as the GAMs initiate anti-tumor response and eradicate residual GBM cells. Taken together, this work provides a potential strategy for immunomodulating the postoperative tumor niche to mitigate tumor relapse in GBM patients, which may have broad applications in other malignancies with surgical intervention.

First Report of Leaf Spot Caused by Alternaria alternata on Chard (Beta vulgaris var. cicla L.) in China.

Chard (Beta vulgaris var. cicla L.) is popular vegetable in China. In June 2023, a leaf spot disease was observed on Chard plants in Hunan Province (27°46'10.99″N, 112°05'52.80″E), China. The disease incidence was 30% in a surveyed of about 500 plants. Symptoms began as many light brown round- to polygon-shaped spots on chard leaves, then developed and enlarged into grayish-white lesions, with the edge of the spots brown to dark brown. A total of 10 symptomatic samples were randomly collected. To identify the pathogen, symptomatic tissues (0.5 × 0.5 cm) from the lesion margin surface were sterilized with 75% ethanol for 30 s and 2% NaClO for 1 min, rinsed 3 times with sterile water, air dried. The sterile pieces were placed on potato dextrose agar (PDA) and incubated at 25°C. A total of nine isolates were obtained. Fungal colonies cultured on potato carrot agar (PCA) were almost the same as each other, and two representative isolates (TC0, TC10) were used for further identification. On PCA, the fungal hyphae were initially white and finally gray-brown with flocculent aerial mycelia. Conidia were solitary or in chains, with various shapes, mostly subglobose, the size was 13.2 to 28.0 μm long and 5.8 to 13.0 μm wide (n = 30). The cultural and morphological characteristics of isolates were similar to those of Alternaria sp (Simmons et al. 2007). For molecular identification, four loci, ITS (White et al. 1990), RPB2 (O'Donnell, 2022), H3 (Zheng et al. 2015), and GAPDH (Berbee et al. 1999), were sequenced from two representative isolates (TC0, TC10). Compared with a reference isolate, Alternaria alternata strain CBS 107.27, GenBank accession nos. KP124300.1 (ITS), KP124768.1 (RPB2), KP124157.1 (GAPDH). The ITS, RPB2, and GAPDH sequences of TC0 and TC10 showed 99% (502 of 504 bp ), 100% (753 of 753 bp), and 99% (560 of 561 bp) similarity, respectively. Compared with a reference isolate, A. alternata isolate 21-5, GenBank accession no. MN840996.1 (H3), H3 sequences of TC0 and TC10 showed 99% (399 of 401 bp) similarity. The sequences of two isolates (TC0, TC10) were deposited in GenBank with accession numbers PP837733.1, PP565404.1(ITS), PP839298.1, PP573905.1(RPB2), PP839299.1, PP573904.1 (GAPDH), and PP839297.1, PP573903.1(H3). Phylogenetic trees were constructed using the sequences and showed that isolates (TC0, TC10) were in the same clade with A. alternata strains. TC0 and TC10 were identified as A. alternata based on the morphological characteristics and molecular phylogeny. Pathogenicity testing was conducted on six-month-old healthy plants, (cv. Green Stalk), three plants were inoculated by spraying spore solution (1 × 106 conidia/mL), and three plants were sprayed with sterile water as a control. The pathogenicity test was performed 3 times. Plants were maintained at 28°C and >80% RH. Plants showed symptoms after 30 days, symptoms were observed similar to those of the original infected plants, control plants were asymptomatic. The fungus was reisolated, confirmed as A. alternata based on conidial characteristics, no pathogenic fungus was isolated from the control plants. A. alternata has previously been reported on beet (also Beta vulgaris) in China (Tai, F. L. 1979; Zhuang, W. Y. 2005). To our knowledge, this is the first report of leaf spot caused by A. alternata on chard in China. This result may expand the etiological study of A. alternata and the control strategy of Chard leaf spot.

Photobiological response of mast cells to green and red light-emitting diodes (LEDs) in cutaneous burns.

This study assessed the effects of red and green LEDs on mast cells (MCs) in third-degree burns in 75 Wistar rats, divided into control, red LED (RED), and green LED (GREEN) groups. Animals were irradiated daily with RED (630 nm, 300 mW, 0.779 W/cm2, 9 J/cm2, 30 s) and GREEN (520 nm, 180 mW, 0.467 W/cm2, 60 J/cm2, 30 s). Histological sections stained with toluidine blue were analyzed for total and subtype MCs. Standardized MC counting was performed across the viable lesion area, considering lesion margins, through intact connective tissue and the integrity of skin appendages. No statistically significant differences in MCs 2 (with released granules and intact cell border) were found between groups. Irradiated groups showed increased total MCs at 7, 14, and 21 days (p < 0.05), with a decrease in MCs 1 (intact MCs) at all time points compared to control (p < 0.05). Significant changes in MCs 3 (with massive degranulation and partial or complete disintegration of the cell border) degranulation were noted in RED at 7, 14, and 21 days (p < 0.009) and in GREEN at 14 (p < 0.009) and 32 days (p < 0.028). Results suggest red and green LEDs modulate MC recruitment and degranulation in third-degree burns.

First report of Erwinia papayae associated with papaya mushy canker disease in Guam.

Carica papaya (papaya) in Guam, USA may experience soft rot symptoms, often referred to as mushy canker disease. Disease symptoms first appear as expanding water-soaked dark-green stem lesions or leaf spotting with chlorotic halos. Defoliation at petiole-stem junctions and crown necrosis leads to plant death. Papaya diseases caused by Erwinia spp. are documented in nearby tropic regions such as the Northern Mariana Islands (Trujillo and Schroth 1982), the Philippines (Dela Cueva et al. 2017), Japan (Hanagasaki et al. 2020), and Indonesia (Suharjo et al., 2021). The pathogen was isolated from symptomatic papaya stem sections (cv. Red Lady) from a nursery at the University of Guam Agriculture and Life Sciences building in April 2023. Approximately 20% of seedlings collapsed from stem soft rot, with nearly all plants showing varying degrees of water-soaked lesions on leaves or stems. Stem tissue from lesion margins was excised, surface sterilized with 70% EtOH, and macerated in sterile water. Macerate was plated onto nutrient agar (NA) and incubated at 28°C, yielding colonies that were clear to white in color, smooth, circular and mucoid on NA plates for five suspect isolates (JGD231-235). Strains produced blue diffusible pigment on King's B (KB) media, were Gram-negative rods, and exhibited swimming motility on semi-solid (0.5% agar) NA plates. Crown stab inoculation of ten papaya plants (cv. Red Lady) with isolates resulted in mushy canker symptoms within seven days, while negative control plants stabbed with a sterile probe remained asymptomatic. Koch's postulates were fulfilled by drench-inoculating spontaneous rifampicin-resistant (100µg/ml) mutants, JGD233r and JGD235r, onto ten papaya plants (cv. Solo Sunrise). Nine days post-inoculation, bacterial strains were recovered from symptomatic stem tissue macerate plated on rifampicin (100µg/ml) NA and incubated at 28°C. No symptoms or bacterial cells were recovered from the tissue of negative control plants. Cell morphology, culture phenotypes, and disease symptoms suggested the isolates were Erwinia spp., and blue pigment production on KB further suggested E. papayae (Gardan et al. 2004). Partial 16S rDNA sequences of Guam strains JGD231-235 (sequenced using PCR forward primer 5' - AGAGTTTGATCMTGGCTCAG - 3' and reverse primer 5' - GGTTACCTTGTTACGACTT - 3', GENEWIZ (South Plainfield, NJ)) were deposited into GenBank (OR577627- 631). Highest NCBI BLAST results for all strains showed a 16S rDNA sequence identity of 98.17-98.91% with those of Erwinia sp. I-leaf (LC590218) and E. mallotivora BT-MARDI (HQ456230). A maximum likelihood phylogenetic tree based on concatenated partial atpD, infB, and rpoB sequences of strains JGD232 (PP669340, PP669346, PP669343), JGD233 (PP669341, PP669347, PP669344), and JGD235 (PP669342, PP669348, PP669345) (Brady et al. 2008) constructed using MEGA11 (Tamura et al. 2021) showed all strains formed a monophyletic group with Erwinia sp. I-leaf (Hanagasaki et al. 2020) and E. papayae NCPPB 4294T (Gardan et al. 2004), supported with 98% bootstrap. This note documents the first occurrence of E. papayae as a papaya pathogen in Guam. Papaya cultivation supports sustainable food security for Guam (Bevacqua and Sayama 2023), and Erwinia spp. pathogens threaten papaya on other Pacific islands like Hawaii. These findings convey the need for effective quarantine practices, local disease management, and further research on this pathogen.

Development and validation of a digital biopsy model to predict microvascular invasion in hepatocellular carcinoma.

Microvascular invasion is a major histopathological risk factor of postoperative recurrence in patients with hepatocellular carcinoma. This study aimed to develop and validate a digital biopsy model using imaging features to predict microvascular invasion before hepatectomy. A total of 217 consecutive patients who underwent hepatectomy for resectable hepatocellular carcinoma were enrolled at two tertiary-care reference centers. An imaging-based digital biopsy model was developed and internally validated using logistic regression analysis with adjustments for age, sex, etiology of disease, size and number of lesions. Three imaging features, i.e., non-smoothness of lesion margin (OR = 16.40), ill-defined pseudocapsula (OR = 4.93), and persistence of intratumoral internal artery (OR = 10.50), were independently associated with microvascular invasion and incorporated into a prediction model. A scoring system with 0 - 3 points was established for the prediction model. Internal validation confirmed an excellent calibration of the model. A cutoff of 2 points indicates a high risk of microvascular invasion (area under the curve 0.87). The overall survival and recurrence-free survival stratified by the risk model was significantly shorter in patients with high risk features of microvascular invasion compared to those patients with low risk of microvascular invasion (overall survival: median 35 vs. 75 months, P = 0.027; recurrence-free survival: median 17 vs. 38 months, P < 0.001)). A preoperative assessment of microvascular invasion by digital biopsy is reliable, easily applicable, and might facilitate personalized treatment strategies.

Conservative Management of Vulvar Cancer-Where Should We Draw the Line?

Vulvar cancer is a rare disease, and cure rates were low until the mid-20th century. The introduction of an en bloc radical vulvectomy and bilateral groin and pelvic lymph node dissection saw them rise from 15-20% to 60-70%. However, this very radical surgery was associated with high physical and psychological morbidity. Wounds were usually left open to granulate, and the average post-operative hospital stay was about 90 days. Many attempts have been made to decrease morbidity without compromising survival. Modifications that have proven to be successful are as follows: (i) the elimination of routine pelvic node dissection, (ii) the use of separate incisions for groin dissection, (iii) the use of unilateral groin dissection for lateral, unifocal lesions, (iv) and radical local excision with 1 cm surgical margins for unifocal lesions. Sentinel node biopsy with ultrasonic groin surveillance for patients with node-negative disease has been the most recent modification and is advocated for patients whose primary cancer is <4 cm in diameter. Controversy currently exists around the need for 1 cm surgical margins around all primary lesions and on the appropriate ultrasonic surveillance for patients with negative sentinel nodes.