Marginal Tissue Research Articles

First Report of Fusarium solani and Fusarium oxysporum Causing Basal Stem Rot on Celery in China.

Celery (Apium graveolens L.) is an economically important vegetable crop in China. In recent years, celery has been widely planted in Yuzhong county, Gansu province. From 11 April to 24 May, 2019-2021, basal stem rot of celery was observed with incidences up to 15% in the Yuzhong region (35°49'N, 104°16'E, 1865 m a.s.l.), which caused serious economic losses to local farmers. Typical symptoms of the disease included wilting and darkening of the basal stem, leading to plant death. To identify the cause of the disease, small pieces (5mm×5mm) of the margin of asymptomatic and rotting basal stem tissues were sterilized with 70% ethanol for 30 s and 3% NaClO for 5 min, then placed on potato dextrose agar (PDA) plates, and incubated at 25℃ (Zhao et al. 2021). Twenty-seven single-conidium isolates with morphological characteristics similar to Fusarium spp. were obtained (Ma et al. 2022), which displayed two kinds of colony morphology. On PDA, seven isolates developed white, fluffy aerial mycelium and twenty isolates developed light pink abundant aerial mycelium. Isolate F5 and F55 from each distinct morphological group were cultured on PDA and synthetic low nutrient agar (SNA) for pathogenicity tests, morphological and molecular identification. The macroconidia (18.3 to 29.6 × 3.6 to 5.3 μm, n=50) with 1 to 2 septa and microconidia (7.5 to 11.6 × 2.6 to 3.5 μm, n=50) with 0 to 1 septum were observed in F5. For F55, the size of macroconidia was 14.2 to 19.5 × 3.3 to 4.2 μm (n = 50) with 1 to 2 septa; Microconidia were mostly 0 to 1 septum and measured 7.3 to 12.8 × 2.2 to 4.2 μm (n = 50). To confirm the identity of the isolates, the internal transcribed spacer region (ITS) and the translation elongation factor-1 alpha (TEF-1α) gene were amplified using primers ITS1/ITS4 and EF-1/EF-2 (Uwaremwe et al. 2020), respectively. The sequence similarities of isolate F5 (GenBank No. OL616048 and OP186480) and F55 (GenBank No. OL616049 and OP186481) with the corresponding sequences of F. solani (MT447508 and MN650097) and F. oxysporum (MG461555 and OQ632904) ranged from 99.22% to 100.00%, with matching base pairs of 531/532, 416/416, 511/515 and 394/395, respectively. The voucher samples were deposited in the sample center of Northwest Institute of Ecological Environment and Resources, Chinese Academy of Sciences. Morphological and molecular results confirmed the species of F5 and F55 as F. solani and F. oxysporum, respectively. A pathogenicity test was conducted under greenhouse conditions (19-31°C, avg. 26°C). Conidial suspension (105 spores/mL) of isolates F5 and F55 were poured onto the basal stems of healthy celery seedlings at the age of 1 month and mock-inoculated control treatments with sterile water. Ten plants were inoculated for each treatment. After 21 days, all plants inoculated with both fungal isolates developed symptoms similar to those observed in the field, whereas the mock-inoculated plants remained healthy. The pathogen was successfully reisolated from the inoculated symptomatic plants onto PDA medium and had morphology as described above, confirming Koch's postulates. F. solani and F. oxysporum have been reported to infect many plant species, including carrot (Zhang et al. 2014) and Angelica sinensis (Liu et al. 2022). To our knowledge, this is the first report that F. solani and F. oxysporum cause basal stem rot on celery in China. The identification of pathogens of the observed basal stem rot on celery provides a clear target for the prevention and management of this disease.

Different patterns of mast cell distribution in B-cell non-Hodgkin lymphomas

Tumor growth, progression, and metastatic capability in non-Hodgkin lymphomas (NHLs) are influenced by different component of tumor microenvironment, including inflammatory cells. Among these latter, mast cells play a crucial role. The spatial distribution of mast cells inside the tumor stroma of different types of B-cell NHLs has not yet been investigated. The aim of this study is to analyze the pattern of distribution of mast cells in biopsy samples obtained from three different types of B-cell NHLs by utilizing an image analysis system and a mathematical model to allow a quantitative estimation to characterize their spatial distribution. As concerns the spatial distributions exhibited by mast cells in diffuse large B cell lymphoma (DLBCL), some clustering was detected in both activated B-like (ABC) and germinal center B-like (GBC) groups. In follicular lymphoma (FL), mast cell spatial distribution tends to uniformly fill the tissue space as far as the grade of the pathology increases. Finally, in marginal lymphoma tissue (MALT) lymphoma, mast cells maintain a significantly clustered spatial distribution, suggesting a lower tendency of the cells to fill the tissue space in this pathological condition. Overall, the data of this study confirm that the analysis of the spatial distribution of the tumor cells is of particular significance for the knowledge of the biological processes occurring in tumor stroma and for the development of parameters to characterize the morphologic organization of the cellular patterns in different types of tumors.

Dental implant and aesthetics: a systematic review

Introduction: Besides the aspects of oral physiological functions, visual appearance and aesthetics have been addressed as an important factors in this type of therapy. In restorative dentistry, a pleasant aesthetic has been described as a blending of natural dentition and prosthetic elements, in addition to the identification, adaptation, and correct conditioning of these elements with the marginal and peripheral tissues. Objective: The present study aimed to present the main factors supporting aesthetic excellence before and after the rehabilitation of former regions' use of dental implants. Methods: The systematic review rules of the PRISMA Platform were followed. The search was carried out from February to May 2023 in the Scopus, PubMed, Science Direct, Scielo, and Google Scholar databases, using articles from 1985 to 2022. The quality of the studies was based on the GRADE instrument and the risk of bias was analyzed according to the Cochrane instrument. Results and Conclusion: A total of 98 articles were found, 38 articles were evaluated in full and 31 were included and developed in this systematic review study. Considering the Cochrane tool for risk of bias, the overall assessment resulted in 10 studies with a high risk of bias and 30 studies that did not meet GRADE. It is important to know the biological structures of the soft and hard tissues around an osseointegrated dental implant (peri-implant referred to as peri-implant phenotype), in the context of peri-implant esthetic complications. Aesthetics has become a primary factor in the patient's expectations, and the duty of the surgeon's professional knowledge of the fundamental aspects in achieving this aspect of a correct treatment plan to meet the appropriate restorative and surgical protocols, thus being able to achieve satisfactory results is required. It notes that all efforts should be aimed at the final result.

Up-regulation of BOK-AS1, FAM215A and FEZF1-AS1 lncRNAs and their potency as moderate diagnostic biomarkers in gastric cancer

BackgroundGastric cancer is the fifth most frequent cancer worldwide and the fourth leading cause of death from cancer, a complex multifactorial neoplasm. LncRNAs are regulatory RNA molecules larger than 200 nucleotides, which can have profound effects on the oncogenic process of various types of cancer. Therefore, these molecules can be used as diagnostic and therapeutic biomarkers. This study aimed to determine the differences in BOK-AS1, FAM215A, and FEZF1-AS1 gene expression between tumor tissue and adjacent healthy non-tumor tissue of gastric cancer (GC) patients. MethodsIn this study one hundred pairs of cancerous and non-cancerous marginal tissues were gathered. Next, RNA extraction and cDNA synthesis were achieved for all of the samples. Then, the qRT-PCR was performed to measure the expression of BOK-AS1, FAM215A and FEZF1-AS1 genes. ResultsAll BOK-AS1, FAM215A and FEZF1-AS1 genes showed significantly increased expression in tumor tissues compared with non-tumor tissues. The outcome of the ROC analysis demonstrated that BOK-AS1, FAM215A, and FEZF1-AS1 may act as mean biomarkers with AUC of 0.7368, 0.7163 and 0.7115, specificity of 64%, 61% and 59%, and sensitivity of 74%, 70%, and 74% respectively. ConclusionBased on the increased expression of the BOK-AS1, FAM215A and FEZF1-AS1 genes in GC patients, this study suggests that these genes may function as oncogenic factors. Furthermore, the mentioned genes can be considered as intermediate biomarkers for diagnosis and treatment of gastric cancer. In addition, no association between these genes and clinicopathological features was observed.

Paired-agent imaging as a rapid en face margin screening method in Mohs micrographic surgery.

Mohs micrographic surgery is a procedure used for non-melanoma skin cancers that has 97-99% cure rates largely owing to 100% margin analysis enabled by en face sectioning with real-time, iterative histologic assessment. However, the technique is limited to small and aggressive tumors in high-risk areas because the histopathological preparation and assessment is very time intensive. To address this, paired-agent imaging (PAI) can be used to rapidly screen excised specimens and identify tumor positive margins for guided and more efficient microscopic evaluation. A mouse xenograft model of human squamous cell carcinoma (n = 8 mice, 13 tumors) underwent PAI. Targeted (ABY-029, anti-epidermal growth factor receptor (EGFR) affibody molecule) and untargeted (IRDye 680LT carboxylate) imaging agents were simultaneously injected 3-4h prior to surgical tumor resection. Fluorescence imaging was performed on main, unprocessed excised specimens and en face margins (tissue sections tangential to the deep margin surface). Binding potential (BP) - a quantity proportional to receptor concentration - and targeted fluorescence signal were measured for each, and respective mean and maximum values were analyzed to compare diagnostic ability and contrast. The BP and targeted fluorescence of the main specimen and margin samples were also correlated with EGFR immunohistochemistry (IHC). PAI consistently outperformed targeted fluorescence alone in terms of diagnostic ability and contrast-to-variance ratio (CVR). Mean and maximum measures of BP resulted in 100% accuracy, while mean and maximum targeted fluorescence signal offered 97% and 98% accuracy, respectively. Moreover, maximum BP had the greatest average CVR for both main specimen and margin samples (average 1.7 ± 0.4 times improvement over other measures). Fresh tissue margin imaging improved similarity with EGFR IHC volume estimates compared to main specimen imaging in line profile analysis; and margin BP specifically had the strongest concordance (average 3.6 ± 2.2 times improvement over other measures). PAI was able to reliably distinguish tumor from normal tissue in fresh en face margin samples using the single metric of maximum BP. This demonstrated the potential for PAI to act as a highly sensitive screening tool to eliminate the extra time wasted on real-time pathological assessment of low-risk margins.

Epipericardial fat necrosis in chest CT and MRI: a case report of an unusual cause of chest pain associated with the initial diagnosis of undifferentiated connective tissue disease

BackgroundEpipericardial fat necrosis (EFN) is a benign and self-limited condition of unknown cause with a good prognosis, usually affecting otherwise healthy patients. Clinically, it presents with severe acute left pleuritic chest pain, often leading the patient to the Emergency Room (ER).Case presentationA 23-year-old male, smoker (5 pack-years), was evaluated in the ER due to left pleuritic chest pain, worsening with deep breathing and Valsalva maneuver. It was not associated with trauma and did not present other symptoms.The physical examination was unremarkable. The arterial blood gases while breathing room air and the laboratory tests, including D-dimers and high-sensitivity cardiac Troponin T, were normal. The chest radiograph, electrocardiogram, and transthoracic echocardiogram showed no abnormalities.A computed tomography (CT) pulmonary angiogram showed no signs of pulmonary embolism but depicted at the left cardiophrenic angle a focal 3 cm ovoid-shaped fat lesion with stranding and thin soft tissue margins, consistent with necrosis of the epicardial fat, which was confirmed by magnetic resonance (MRI) of the chest.The patient was medicated with ibuprofen and pantoprazole, with clinical improvement in four weeks. At a two-month follow-up, he was asymptomatic and presented radiologic resolution of the inflammatory changes of the epicardial fat of the left cardiophrenic angle on chest CT. Laboratory tests revealed positive antinuclear antibodies, positive anti-RNP antibody, and positive lupus anticoagulant. The patient complained of biphasic Raynaud’s phenomenon initiated five years ago, and a diagnosis of undifferentiated connective tissue disease (UCTD) was made.ConclusionsThis case report highlights the diagnosis of EFN as a rare and frequently unknown clinical condition, which should be considered in the differential diagnosis of acute chest pain. It can mimic emergent conditions such as pulmonary embolism, acute coronary syndrome, or acute pericarditis. The diagnosis is confirmed by CT of the thorax or MRI. The treatment is supportive and usually includes non-steroidal anti-inflammatory drugs. The association of EFN with UCTD has not been previously described in the medical literature.

Does joint-sparing tumor resection jeopardize oncologic and functional outcomes in non-metastatic high-grade osteosarcoma around the knee?

BackgroundWe previously reported joint-sparing tumor resection for osteosarcoma with epiphyseal involvement in which transepiphyseal osteotomy went through the in situ ablated epiphysis. However, we do not know whether this is a safe approach when compared with joint-sacrificed tumor resection. Our objective was to compare oncologic and functional outcomes between patients who underwent joint preservation (JP) and joint replacement (JR) tumor resection. Furthermore, we identified the risk factors of local recurrence, metastasis and survival.MethodsEighty-nine patients with non-metastatic high-grade osteosarcoma around the knee were treated with limb-salvage surgery (JP in 47 and JR in 42). Age, gender, tumor location, pathologic fracture, plain radiographic pattern, limb diameter change, perivascular space alteration, surgical margin, local recurrence, metastasis, death, and the Musculoskeletal Tumor Society (MSTS)-93 scores were extracted from the records. Univariate analysis was performed to compare oncologic and functional outcomes. Binary logistic and cox regression models were used to identify predicted factors for local recurrence, metastasis, and survival.ResultsLocal recurrence, metastasis and overall survival were similar in the JP and JR group (p = 0.3; p = 0.211; p = 0.143). Major complications and limb survival were also similar in the JR and JP group (p = 0.14; p = 0.181). The MSTS score of 27.06 ± 1.77 in the JP group was higher than that of 25.88 ± 1.79 in the JR group (p = 0.005). The marginal margin of soft tissue compared with a wide margin was the only independent predictor of local recurrence (p = 0.006). Limb diameter increase and perivascular fat plane disappearance during neoadjuvant chemotherapy were independent predictors for metastasis (p = 0.002; p = 0.000) and worse survival (p = 0.000; p = 0.001).ConclusionsJoint-sparing tumor resection with the ablative bone margin offers advantage of native joint preservation with favorable functional outcomes while not jeopardizing oncologic outcomes compared with joint-sacrificed tumor resection. Surgeon should strive to obtain adequate soft tissue surgical margin decreasing risk of local recurrence. Novel drug regimens might be reasonable options for patients with obvious limb diameter increase and perivascular fat disappearance during chemotherapy.

Imaging guidance for cholesteatoma surgery using tissue autofluorescence.

Cholesteatoma is an expansile destructive lesion of the middle ear and mastoid, which can result in significant complications by eroding adjacent bony structures. Currently, there is an inability to accurately distinguish cholesteatoma tissue margins from middle ear mucosa tissue, causing a high recidivism rate. Accurately differentiating cholesteatoma and mucosa will enable a more complete removal of the tissue. Develop an imaging system to enhance the visibility of cholesteatoma tissue and margins during surgery. Cholesteatoma and mucosa tissue samples were excised from the inner ear of patients and illuminated with 405, 450, and 520nm narrowband lights. Measurements were made with a spectroradiometer equipped with a series of different longpass filters. Images were obtained using a red-green-blue (RGB) digital camera equipped with a long pass filter to block reflected light. Cholesteatoma tissue fluoresced under 405 and 450nm illumination. Middle ear mucosa tissue did not fluoresce under the same illumination and measurement conditions. All measurements were negligible under 520nm illumination conditions. All spectroradiometric measurements of cholesteatoma tissue fluorescence can be predicted by a linear combination of emissions from keratin and flavin adenine dinucleotide. We built a prototype of a fluorescence imaging system using a 495nm longpass filter in combination with an RGB camera. The system was used to capture calibrated digital camera images of cholesteatoma and mucosa tissue samples. The results confirm that cholesteatoma emits light when it is illuminated with 405 and 450nm, whereas mucosa tissue does not. We prototyped an imaging system that is capable of measuring cholesteatoma tissue autofluorescence.

B7H3 Role in Reshaping Immunosuppressive Landscape in MSI and MSS Colorectal Cancer Tumours.

The study aimed to assess the expression of B7H3 concerning clinicopathological and histological parameters, including MSI/MSS status, CD-8 cells, tumour-infiltrating lymphocytes (TILs), budding, TNM scale and grading. Moreover, we analyzed the B7H3-related pathways using available online datasets and the immunological context of B7H3 expression, through the 48-cytokine screening panel of cancer tissues homogenates, immunogenic features and immune composition. The study included 158 patients diagnosed with CRC. To assess B7H3 levels, we performed an immunohistochemistry method (IHC) and enzyme-linked immunosorbent assay (ELISA). To elucidate the immune composition of colorectal cancer, we performed the Bio-Plex Pro Human 48-cytokine panel. To study biological characteristics of B7H3, we used online databases. Expression of B7H3 was upregulated in CRC tumour tissues in comparison to adjacent noncancerous margin tissues. The concentrations of B7H3 in tumours were positively associated with T parameter of patients and negatively with tumour-infiltrating lymphocytes score. Additionally, Principal Component Analysis showed that B7H3 expression in tumours correlated positively with cytokines associated with M2-macrophages and protumour growth factors. The expression of B7H3 in tumours was independent of MSI/MSS status. These findings will improve our understanding of B7H3 role in colorectal cancer immunity. Our study suggests that B7-H3 is a promising potential target for cancer therapy. Further studies must clarify the mechanisms of B7H3 overexpression and its therapeutic importance in colorectal cancer.

First report of Neopestalotiopsis clavispora Causing Leaf Spots on Garcinia mangostana in China.

Garcinia mangostana L. is a famous tropical fruit in Asia. In April 2021, a leaf disease on G. mangostana cv. Huazhu was observed in Zhanjiang (21.17° N, 110.18° E), Guangdong province, China. Symptoms was on new leaves of 2 year old plants. The spots were circular to irregular, gray in the center, and brown on the lesion margin. The disease incidence was estimated 25% (n = 500 investigated plants from about 50-ha). Twenty diseased leaves were collected from the orchard. The margin of the diseased tissues was cut into 2 mm × 2 mm pieces; surface disinfected with 75% ethanol and 2% sodium hypochlorite for 30 and 60 s, respectively; and rinsed thrice with sterile water. The tissues were plated onto potato dextrose agar (PDA) medium and incubated at 28 ℃. Twenty-eight isolates were obtained (isolation frequency = 28/4×20 = 35%). Single-spore isolation method was used to recover pure cultures for three isolates (GMN-1, GMN-2, and GMN-3) (Liu et al. 2021). The colonies were initially white with cottony aerial mycelium at 7 days on PDA. Then, they developed black acervular conidiomata at 10 days. Conidia were clavate to fusiform, four-septate, straight or slightly curved, and measured 16.5 to 21.4 µm long (average 19.5 µm; n = 40) × 4.5 to 6.5 µm wide (average 5.2 µm; n = 40). The three median cells were versicolored, whereas the basal and apical cells were hyaline. Conidia had a single basal appendage (4.5 to 5.5 µm long; n = 40) and three apical appendages (19.2 to 24.5 µm long; n = 40). The morphological characteristics of the isolates are comparable with those of the genus Neopestalotiopsis(Sajeewa et al. 2012). Molecular identification was performed using the colony polymerase chain reaction method with MightyAmp DNA Polymerase (Takara-Bio, Dalian, China) (Lu et al. 2012). Sequences were generated from the isolates using primers for the rDNA ITS (ITS1/ITS4), TEF1-α (EF1-728F/EF1-986R), and β-tubulin (T1/βt2b) loci (Sajeewa et al. 2012). The sequences of the isolates were submitted to GenBank (ITS, MZ026535-MZ026537; TEF, MZ032203-MZ032205; β-tubulin, MZ032206-MZ032208). The sequences of the isolates were 100% identical to the type strain MFLUCC12-0281 (accession nos. JX398979, JX399014, and JX399045) through BLAST analysis. The isolates clustered with N. clavispora (MFLUCC12-0280 and MFLUCC12-0281). The pathogenicity was tested in vivo. Individual plants (cv. Huazhu) were grown (n = 2, 1-1.5 year old) in a greenhouse at 24℃-30℃ with 80% relative humidity. Wounded leaflets were inoculated with 5-mm-diameter mycelial plugs or agar plugs (as control). Besides, sterile cotton balls were immersed in the spore suspension (1 × 105 per mL) and sterile distilled water (control) for about 15 s before they were fixed on the leaves for 3 days. One plant employed for each isolate with nine leaves. The test was performed thrice. Disease symptoms were found on the leaflets after 10 days, whereas the controls remained healthy. The pathogen was re-isolated from infected leaves and phenotypically identical to the original isolates to fulfill Koch's postulates. Neopestalotiopsis clavispora and Pestalotiopsis clavispora are synonyms. The fungus appeared to have a wide host range and distribution including in Thailand, Malaysia, North Queensland, and Australia (Sajeewa et al. 2012;Shahriar et al. 2022). Thus, this is the first report of N. clavispora causing leaf spot on G. mangostana in China. This finding will help improve management strategies against the leaf spots on G. mangostana in China.

First Report of Leaf Spot and Stem Rot Caused by Diaporthe eres on Hydrangea macrophylla in Italy

HomePlant DiseaseAhead of PrintFirst Report of Leaf Spot and Stem Rot Caused by Diaporthe eres on Hydrangea macrophylla in Italy PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Leaf Spot and Stem Rot Caused by Diaporthe eres on Hydrangea macrophylla in ItalyA. Garibaldi, G. Tabone, I. Luongo, and M. L. GullinoA. GaribaldiCentre of Competence AGROINNOVA, University of Torino, 10095 Grugliasco, ItalySearch for more papers by this author, G. TaboneCentre of Competence AGROINNOVA, University of Torino, 10095 Grugliasco, ItalySearch for more papers by this author, I. LuongoCentre of Competence AGROINNOVA, University of Torino, 10095 Grugliasco, ItalySearch for more papers by this author, and M. L. Gullino†Corresponding author: M. L. Gullino; E-mail Address: [email protected]https://orcid.org/0000-0002-7706-1915Centre of Competence AGROINNOVA, University of Torino, 10095 Grugliasco, ItalySearch for more papers by this authorAffiliationsAuthors and Affiliations A. Garibaldi G. Tabone I. Luongo M. L. Gullino † Centre of Competence AGROINNOVA, University of Torino, 10095 Grugliasco, Italy Published Online:9 Jun 2023https://doi.org/10.1094/PDIS-08-22-1786-PDNAboutSectionsPDF ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Native to the temperate and subtropical regions of Japan and China, Hydrangea macrophylla (big leaf hydrangea) is a shrubby plant in the Hydrangeaceae used to embellish parks, gardens, private terraces, and balconies. Punctiform brown leaf spots and stem necrosis appeared in the summer of 2021 on 10 of approximately 50 H. macrophylla plants. The 20-year-old plants were cultivated in pots and in mixed borders in a private garden located in Piedmont (Biella Province 45°36′00″N, 8°03′00″E). Spots enlarged to encompass the entire surface; eventually affected leaves abscised. Affected leaves at the early stage of disease were randomly collected and fragments, obtained from the margins of necrotic tissues, were disinfected in 1% sodium hypochlorite, rinsed in sterilized water, and distributed on potato dextrose agar (PDA) amended with 25 mg/liter of streptomycin sulfate. Plates were maintained at room temperature (22 to 26°C) and colonies, characterized by white, aerial, fluffy mycelium with a dark pigmentation at the center, appeared after 5 days. Mycelium of one representative pure culture grown on PDA for 20 days became gray with age and developed dark brown pycnidia and black stromata at maturity. Alpha conidia were hyaline, aseptate, fusiform to ellipsoidal, often biguttulate, and 5.1 to 10.2 (average 7.2) × 1.9 to 3.7 (average 3.0) μm (n = 50). Beta conidia were hyaline, aseptate, linear to curved, and 14.5 to 35.7 (average 24.9) × 0.6 to 2.9 (average 1.6) μm (n = 50). These morphological characteristics permitted identification of the fungus as Diaporthe sp. (Gomes et al. 2013). Polymerase chain reaction (PCR) and Sanger sequencing was carried out for ITS, calmodulin (cal) (Cal228F/Cal2Rd), tef1 (EF1-728/EF1-986R), and tub2 (T1/Bt2b) (Udayanga et al. 2014) of the representative isolate 22/12-1 (GenBank accessions OP080713, OP700046, OP14610, and OP700047, respectively). Sequences analyzed with BLASTn (Altschul et al. 1997) showed 99.5% identity for ITS (567/570 bp; KC343097), 99.8% for cal (414/415 bp; KC343339), 100% for tef1 (347/347 bp; KC343823), and 100% for tub2 (690/690 bp; KC344065) with sequences from strain CBS 841.84 of Diaporthe eres. Pathogenicity of the isolate 22/12-1 was verified by performing a pathogenicity test on five healthy 20-year-old H. macrophylla plants grown in pots and in borders. A conidial suspension at a concentration of 1 × 105 conidia/ml, obtained from a pure culture of the isolate grown for 20 days on potato dextrose agar (PDA), was sprayed on the leaves of the H. macrophylla plants (5 ml/plant). Three noninoculated plants sprayed with sterile deionized water were used as a control. Plants were covered with a transparent plastic bag and maintained in the garden at a temperature of 24 to 36°C. Leaf spots and stem necrosis developed on the inoculated plants after 7 to 10 days. Colonies with the same morphological characteristics of the original isolate were isolated from the margins of the necrotic leaf tissue. The pathogenicity test was repeated once, obtaining the same results. D. eres was previously reported on several species of Hydrangea (Farr and Rossman 2022). To our knowledge this is the first report of D. eres on H. macrophylla in Italy. Diaporthe (syn. Phomopsis) is the causal agent of severe disease and could represent a serious problem for the cultivation of H. macrophylla in gardens and a reduction of the ornamental value.The author(s) declare no conflict of interest.

First report of leaf blight caused by Alternaria alternata on Platanus acerifolia in China.

Platanus acerifolia Willd. is widely planted in cities in China due to its strong adaptability to different environmental conditions. In August 2021, light brown, oval to circular, sunken spots were observed on leaves of P. acerifolia trees with 8-35% incidence, leading to severe necrosis and abscission of leaves on a street in Haidian district of Beijing (116°29'84''E, 39°95'93''N). Small pieces (5 mm×5 mm) were taken from the margin of diseased tissues, disinfected with 0.3% sodium hypochlorite for 2 min and 70% ethanol for 40 s, rinsed with sterile water, then plated on potato dextrose agar (PDA) and incubated at 28°C. After 4 days, representative isolates were transferred to new PDA plates. Four isolates with similar morphological characteristics were obtained and deposited in the culture collection (ID: DAA3, DAA5, DAA6 and DAA7) of our laboratory. Colonies on PDA were dense, fluffy, and light to dark gray, with a prominent white margin. Conidia formed in chains on the branched conidiophores, and were obpyriform to ellipsoid, 19.5-32.3×5.5-10.2 μm (average=26.4×7.1 µm, n=30) in size, with 3 to 5 transversal and 1 to 3 longitudinal septa. These morphological characteristics matched those of Alternaria spp. (Simmons 2007). Genomic DNA was extracted with modified CTAB method and the internal transcribed spacer (ITS) region, translation elongation factor 1-α (EF1-α), RNA polymerase II largest subunit (RPB2), glyceraldehyde 3 - dehydrogenase (GPD), endopolygalacturonase (EndoPG), as well as Alternaria major allergen (Alt a1) genes were amplified with primer pairs ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), RPB2-5F/RPB2-7cR (Liu et al. 1999), gpd1/gpd2 (Berbee et al. 1999), PG3/PG2b (Andrew et al. 2009), and Alt-for/Alt-rev (Hong et al. 2005), respectively. The obtained sequences were deposited in GenBank (accession numbers: OM228653-OM228656, OM221523-OM221542). In a BLAST search, the sequences were 100% identical with corresponding sequences of A. alternata. Phylogenetic analysis based on combined sequences using maximum parsimony method showed that the four isolates clustered together with the type strain CBS 916.96 of A. alternata. For pathogenicity test, three healthy leaves of three one-year-old P. acerifolia plants were wounded with a sterile needle and inoculated with 20 µl of spore suspension (106 conidia/ml). Plants inoculated with sterile water were treated as control. The pathogenicity test was also conducted on the unwounded leaves. After 8 days of inoculation at 25°C and 90% RH with a 12-h photoperiod, the symptoms on spore suspension-inoculated leaves were similar to those observed on trees in the street, whereas the control leaves remained symptomless. Lesions on wounded leaves were larger than those on unwounded leaves. The assay was repeated twice with consistent results. The pathogen was re-isolated from symptomatic leaf tissues and identified based on morphological and rDNA-ITS sequencing, thus, fulfilling Koch's postulates. Examples of other tree species where Alternaria alternata has been reported to cause leaf blight were Ophiopogon japonicas in China (Wang et al. 2021) and Pistacia terebinthus in Spain (López-Moral et al. 2018). To our knowledge, this is the first report of A. alternata causing leaf blight on P. acerifolia in China. The identification could provide information for developing effective disease management strategies.

Identifying the driver histology responsible for the inferior clinicopathologic features associated with plasmacytoid bladder cancer.

e16505 Background: Plasmacytoid urothelial cancer (PUC) is notorious for its spread along fascial planes which reportedly induces worse clinicopathologic. Usually, PUC is present with Conventional UC (CUC); however, the precise assessment of the PUC component in key pathological findings and its effect on clinical behavior has not been delineated. Methods: We performed a retrospective review of patients with PUC who underwent cystectomy at Moffitt Cancer Center between 2012-2022. Repeat histological review of all specimens by specialty trained genitourinary pathologists was performed (AV, ED). Positive margins and lymph nodes were examined and classified by histological subtype. We calculate the proportion of tissue sections with peri-vesical fat invasion in tumor tissue sections submitted from cystectomy specimen in pT3 cases as the microscopic surrogate for macroscopic pT3 sub staging (termed Peri-vesical Fat Invasion Ratio, PFIr). Pearson’s correlation coefficient was used to measure strength and direction of relationship between variables. Kaplan-Meier (KM) analysis was performed for overall survival (OS). Results: Population consists of 44 patients undergoing cystectomy for PUC. Median age was 70 (IQR 63-75) years old with 82% being male. Majority of patients (52.5%) had ≥cT3 disease and 55% received neoadjuvant chemotherapy (NAC). Median percent of plasmacytoid histology was 20% (IQR 5-80%) with 6 (14%), 2 (5%), 5 (11%), 16 (36%), and 15 (34%) being pT0, pT1, pT2, pT3, and pT4, respectively. Of the patients with pT3 disease, 6 were pT3a with a median 78% (IQR 39-82%) PFIr while 10 were pT3b with an associated median PFIr of 56% (42-63%). In total, 14 (32%) patients had positive soft tissue surgical margin, with 11 (79%) being of PUC histology and ten patients with a positive ureteral margin, 80% of which consisted of PUC histology. A total of 97 lymph nodes removed were positive for malignancy with 51.5% demonstrating PUC and 48.5% CUC histology. Univariate analysis confirmed an increasing percentage of PUC histology portended a worse OS (HR 3.27, 95% CI 1.22-8.73, p = 0.018). Additionally, we calculated Pearson’s correlation coefficient for percent of plasmacytoid histology with pathologic tumor stage (r = 0.468, p = < 0.001), soft tissue margin (r = 0.283, p = 0.031), and lymph node density (r = 0.264, p = 0.138). Finally, KM analysis for histologic subtype (PUC vs CUC) at positive margin or positive lymph did not affect OS (p > 0.05), likely a product of our small numbers. Conclusions: The plasmacytoid component of bladder cancer is more likely to spread through direct extension along tissue planes to the surrounding soft tissue and ureters rather than through lymphatics. Also, given the discohesive nature of PUC histology, gross peri-vesical extension may be an inadequate method of sub-stratifying stage pT3 disease.

Loss of Caveolin-1 Expression in Tumor Cells is Associated with Increased Aggressiveness and Cell Invasion in Oral Squamous Cell Carcinoma.

Changes in Caveolin-1 (CAV-1) expression are related to tumorigenesis. The aim of this study was to evaluate the role of CAV-1 in tumor progression in oral squamous cell carcinoma (SCC) tissue samples and the effect of CAV-1 silencing on two oral tongue SCC (OTSCC) cell lines (SCC-25, from a primary tumor, and HSC-3 from lymph node metastases). Mycroarray hybridization, mRNA expression, and immunohistochemistry were performed on OSCC tissue samples and corresponding non-tumoral margin tissues. The effects of CAV-1 silencing (siCAV-1) on cell viability, membrane fluidity, on the expression of epithelial to mesenchymal transition (EMT) markers and on cell migration and invasion capacity of OTSCC cell lines were evaluated. Microarray showed a greater CAV-1 expression (1.77-fold) in OSCC tumors than in non-tumoral tissues and 2.0-fold more in less aggressive OSCCs. However, significant differences in CAV-1 gene expression were not seen between tumors and non-tumoral margins nor CAV-1 with any clinicopathological parameters. CAV-1 protein was localized both in carcinoma and in spindle cells of the tumor microenvironment (TME), and CAV-1 positive TME cells were associated with smaller/more aggressive tumors, independent of the carcinoma cells' expression. Silencing of CAV-1 increased cell viability only in SCC-25 cells. It also stimulated the invasion of HSC-3 cells and increased ECAD and BCAT mRNA in these cells; however, the protein levels of the EMT markers were not affected. Decreased expression of CAV-1 by tumor cells in OSCC and an increase in the TME were associated with increased cell invasiveness and tumor aggressiveness.

The Synergistic Mechanism of Photosynthesis and Antioxidant Metabolism between the Green and White Tissues of Ananas comosus var. bracteatus Chimeric Leaves.

Ananas comosus var. bracteatus (Ac. bracteatus) is a typical leaf-chimeric ornamental plant. The chimeric leaves are composed of central green photosynthetic tissue (GT) and marginal albino tissue (AT). The mosaic existence of GT and AT makes the chimeric leaves an ideal material for the study of the synergistic mechanism of photosynthesis and antioxidant metabolism. The daily changes in net photosynthetic rate (NPR) and stomatal conductance (SCT) of the leaves indicated the typical crassulacean acid metabolism (CAM) characteristic of Ac. bracteatus. Both the GT and AT of chimeric leaves fixed CO2 during the night and released CO2 from malic acid for photosynthesis during the daytime. The malic acid content and NADPH-ME activity of the AT during the night was significantly higher than that of GT, which suggests that the AT may work as a CO2 pool to store CO2 during the night and supply CO2 for photosynthesis in the GT during the daytime. Furthermore, the soluble sugar content (SSC) in the AT was significantly lower than that of GT, while the starch content (SC) of the AT was apparently higher than that of GT, indicating that AT was inefficient in photosynthesis but may function as a photosynthate sink to help the GT maintain high photosynthesis activity. Additionally, the AT maintained peroxide balance by enhancing the non-enzymatic antioxidant system and antioxidant enzyme system to avoid antioxidant damage. The enzyme activities of reductive ascorbic acid (AsA) and the glutathione (GSH) cycle (except DHAR) and superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were enhanced, apparently to make the AT grow normally. This study indicates that, although the AT of the chimeric leaves was inefficient at photosynthesis because of the lack of chlorophyll, it can cooperate with the GT by working as a CO2 supplier and photosynthate store to enhance the photosynthetic ability of GT to help chimeric plants grow well. Additionally, the AT can avoid peroxide damage caused by the lack of chlorophyll by enhancing the activity of the antioxidant system. The AT plays an active role in the normal growth of the chimeric leaves.

HIF-1α gene/protein and oxidative stress in patients with colorectal cancer: A pilot study

BackgroundSeveral risk factors such as may enhance the development of colorectal cancer (CRC). These include hypoxia and oxidative stress. We investigated serum pro-oxidant antioxidant balance (PAB) and HIF-1α Gene/protein levels in CRC patients over 3 months. MethodFifty newly diagnosed cases of CRC and 50 healthy individuals were recruited. Tumor sections and marginal healthy tissue, for use as control tissue, were obtained on the day of surgery and HIF-1α mRNA gene expression was determined using Real-Time PCR. Blood from each patient was collected to measure serum PAB using a colorimetric assay, and serum HIF-1α protein concentrations were measured using ELISA method in the patient group before, 24 h after surgery, at the time of discharge, one month, and three months after surgery. The control group samples were obtained on the day of a routine check-up. ResultsThe expression of the HIF-1α gene in the tumor tissue was 7.31 times greater than in healthy tissue margins (p < 0.001) and the level of expression was significantly higher in men than woman (p = 0.006). The mean serum of HIF-1α protein levels and PAB values in patients significantly decreased after surgery up until 3 months (p < 0.001 and p < 0.001; respectively), and were not significantly different between the two genders during follow up (p > 0.05). ConclusionHIF-1α gene expression was significantly higher in tumor tissue than normal marginal tissue and serum HIF-1α protein and PAB levels in patients with CRC were higher preoperatively than the control subjects, and decreased over the 3 months of follow-up.

Comparison of the clinical and prognosis risk factors between endoscopic resection and radical gastrectomy for early-stage gastric cancer

AimThis study aimed to explore the efficacy and safety of endoscopic submucosal dissection/endoscopic mucosal resection (ESD/EMR), laparoscopic-assisted radical gastrectomy (LARG), and open radical gastrectomy (ORG) in early-stage gastric cancer.MethodsA total of 417 patients with early-stage gastric cancer who were admitted in two hospitals from January 1, 2014 to July 31, 2017 were selected; the patients were divided into the ESD/EMR group (139 cases), LARG group (108 cases), and ORG group (170 cases) according to the operation methods used. The baseline data, economic cost of health, oncologic characteristics, postoperative complications, 5-year overall survival and disease-free survival, and risk factors of death were compared and analyzed.ResultsNo significant difference was observed in the baseline data among the three patient groups (P > 0.05). The total hospitalization days, operation time, postoperative fluid intake time, hospitalization expenses, and proportion of antibiotic use rate in the ESD/EMR group were lesser than those in other groups (P < 0.05). The LARG group has a longer operation time and higher hospitalization expenses compared with the ORG group (P < 0.05), but the total hospitalization days, postoperative fluid intake time, proportion of antibiotic use, and lung infection status were consistent. The ESD/EMR group had a lower incidence of incision site infection and postoperative abdominal distension compared with that of the surgery groups (P < 0.05). Five patients required radical surgery after undergoing ESD/EMR (The patients had residual tissue margin cancer), while none of the patients had switched to ORG during LARG. Surgery had advantages over ESD/EMR in terms of lymph node dissection (P < 0.05). No significant differences were observed in the postoperative complications such as upper gastrointestinal bleeding, perforation, incision hernia, reoperation and recurrence (P > 0.05). The 5-year postoperative survival rates of patients in the three groups were 94.2% (ESD/EMR), 93.5% (LARG), and 94.7% (ORG), respectively, with no significant differences (P > 0.05). The binary logistics multivariate analysis showed that the tumor size, invasion depth, vascular invasion, and differentiated degree were risk factors for death in patients with gastric cancer.ConclusionsNo significant difference was observed between ESD/EMR and radical surgery. However, standardized criteria for excluding metastatic lymph nodes should be established to promote ESD/EMR.

Evaluation of the TXNIP gene expression in gastric adenocarcinoma and its relationship with overall survival of patients

Background. Gastric Cancer (GC) is a common gastrointestinal tumor, and its incidence is increasing. The pathogenesis of GC is very complex and remains unclear. Recent basic studies of GC have focused on gene expression dysregulation. Accumulating evidence has demonstrated that thioredoxin-interacting protein (TXNIP) is abnormally expressed in a variety of malignant tumors but it is obscure in GC. This study aimed to compare the expression of the TXNIP gene in cancerous and adjacent tissue of gastric cancer and evaluate its clinical significance in the prognosis and survival analysis of gastric cancer patients. Methods. A total of 50 tumor tissues and marginal non-tumor control tissues were obtained from GC patients. Also, TXNIP gene expression levels were evaluated by real-time PCR. Meanwhile, bioinformatic approaches were used to evaluate TXNIP expression in two different cohorts of GC patients. A data mining study was also performed to determine the prognostic role of TXNIP expression in the overall survival of GC. Furthermore, a pan-cancer analysis of TXNIP expression was performed using TCGA data. Results. The TXNIP gene was down-regulated in tumor samples with a fold change of 0.33, and the same results were repeated in bioinformatics analysis. Decreased expression of TXNIP was significantly associated with metastasis, poor differentiation, and drug abuse. Our results provided evidence that TXNIP gene expression level is positively correlated with the overall survival of GC patients. Pan-cancer analysis of TCGA data revealed down-regulated TXNIP in a variety of malignant tumors. Conclusion. This study established low TXNIP expression as a prognostic biomarker in GC. It also revealed that the decrease in TXNIP expression likely favors metastatic GC. Practical Implications. Evaluation of TXNIP expression is informative in the prognosis of GC patients.

NIRDye 812: A molecular platform tailored for multimodal bioimaging applications of targeted fluorescence- and photoacoustic-guided surgery.

The primary treatment for malignant tumors remains to be surgical removal of the diseased tissue. The presence or absence of residual diseased tissue at the tumor margin is the strongest predictor of postoperative prognosis and recurrence. Accordingly, reliance on the ability of surgeons to visually distinguish diseased tissue from healthy tissue unambiguously in real time is crucial. Near infrared-I (NIRI) fluorescence-emitting targeting biomolecular constructs such as anticancer antibody-fluorophore conjugates, namely cetuximab-IRDye® 800CW (CTB-IRDye® 800CW), are FDA-approved for clinical trial usage in the fluorescence-guided resection of diseased tissue due to affording improved direct visualization of tumor tissue when compared to the use of either the unaided eye under standard white light illumination (WLI) surgical techniques or non-targeting fluorophores. Unfortunately, though helpful, CTB-IRDye® 800CW affords limited (i) identification of diseased tissue and (ii) tumor margin delineation, because the immunoconjugate generates suboptimal tumor-to-background ratios (TBRs) as a result of its spectral/photophysical profiles poorly aligning with the fixed optical windows of pre-/clinical setups. As such, CTB-IRDye® 800CW is more prone to affording incomplete resection compared to if TBRs were higher due to otherwise. To aid in accurately identifying deep-seated diseased tissue, photoacoustic (PA) tomography has been implemented alongside CTB-IRDye® 800CW to achieve PA signals that could result in higher TBRs. However, in clinical trial practice, using IRDye® 800CW for PA imaging also yields subpar TBRs due to it affording low PA signals. To overcome such limitations, we developed NIRDye 812, a structurally-modified topological equivalent of IRDye® 800CW, to confer it the capability to yield both higher TBRs and superior PA signal than that of the equivalent CTB-conjugate and fluorophore IRDye® 800CW itself, respectively. To do so, we substituted the oxygen atom at its meso-position with a sulfur atom. CTB-NIRDye 812 demonstrated a red-shifted absorption wavelength at 796nm and a peak NIR-I fluorescence emission wavelength at 820nm, which better dovetails with the fixed windows of preinstalled fixed emission filters within commercial pre-/clinical NIR-I fluorescence imaging instruments. Overall, CTB-NIRDye 812 provided a∼2-fold increase in TBRs compared to those of CTB-IRDye® 800CW in vivo. Also, NIRDye 812 displayed an ∼60% higher PA signal than that of IRDye® 800CW. Collectively, we achieved our goal of improving upon the spectral/photophysical and PA properties of IRDye® 800CW via introducing a subtle modification to its electronic core such that its CTB immunoconjugate could potentially allow for fast track or breakthrough designation by the FDA due to its near-identical structure displaying considerably improved efficacy.

First Report of Anthracnose on Chili Pepper Caused by Colletotrichum sojae in Hebei Province, China.

China is the largest chili pepper producing country, and Hebei Province stands out as the forth with planting area at about 1500 km2 in China. Pepper (Capsicum annuum L.) is susceptible to Colletotrichum spp. infection during its growth, which seriously affects production yield and quality. In September 2020, widespread anthracnose was observed on pepper in Hebei (115.48° N, 38.77° E), China. Necrotic lesions on pepper fruits were suborbcular, sunken, with acervuli arranged in the middle of lesion (e-Xtra 1A). To perform fungal isolation, small tissue with 0.3 cm2 in size at the symptomatic tissue margin was surface disinfested with 75% ethanol for 10 s, and 0.1% HgCl2 for 40 s, then washed three times with sterile ddH2O. Fragments were placed on potato dextrose agar (PDA) amended with 100 mg·L-1 chloramphenicol and incubated at 28 ºC under darkness for 4 days. One of the strains of Colletotrichum spp., named HQY157, was purified by single-spore isolation, then used for morphological characterization, phylogenetic analysis, and pathogenicity tests. Colonies presented light grey aerial mycelium, occasionally mixed with gray-black strips, and the reverse was similar to the surface on PDA (e-Xtra 1B). Conidia were smooth-walled, aseptate, straight with obtuse to slightly rounded ends, 17.3-28.5 × 3.1-7.4 μm (n=50) (e-Xtra 1C). For molecular identification, the internal transcribed spacer (ITS) region, partial sequences of actin (ACT), β-tublin (TUB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and chitin synthase (CHS) were sequenced using the specific primers (Weir et al. 2012). Sequences were deposited in GenBank with the following accession numbers OM317600-OM317604. A Maximum-Likelihood phylogenetic tree was constructed, based on the concatenated sequences (ACT, CHS, GAPDH, TUB, and ITS) of HQY157 and other closely matching Colletotrichum species obtained from GenBank, by using MEGA-X. It showed that HQY157 was grouped with the C. sojae with bootstrap values of 100% (e-Xtra 2). To confirm the pathogenicity, surface-sterilized healthy pepper fruits and healthy fruits with wounds (deal with a sterile toothpick after surface-sterilized) were then inoculated with 2 μL of conidial suspension (106 conidia/mL). The fruits inoculated with 2 μL sterile distilled water were taken as negative controls. After inoculation, the fruits were kept in a plastic box with sterilized filter paper moistened with sterilized water, and maintained at 25°C in the dark. The experiment was repeated three times. Anthracnose symptoms were observed 7 days after inoculation on the wounded pepper fruits, whereas the unwounded and negative control fruits remained symptomless (e-Xtra 1D). Colletotrichum sojae was re-isolated from the infected pepper fruits and identified by morphological and molecular analysis, fulfilling Koch's postulates. Colletotrichum sojae occurs mainly on Fabaceae plants such as Glycine max, Medicago sativa, Phaseolus vulgaris, and Vigna unguiculata (Damm et al. 2019, Talhinhas and Baroncelli, 2021), and Panax quinquefolium (Guan et al. 2021). To our knowledge, this is the first report of C. sojae causing anthracnose on pepper in China. This study provided crucial information for epidemiologic studies and appropriate control strategies for this chili pepper disease.