Spatial proteo-transcriptomic profiling reveals the molecular landscape of borderline ovarian tumors and their invasive progression.

Serous borderline tumors (SBT) are epithelial neoplastic lesions of the ovaries that commonly have a good prognosis. In 10-15% of cases, however, SBT will recur as low-grade serous cancer (LGSC), which is deeply invasive and responds poorly to current standard chemotherapy1,2,3. While genetic alterations suggest a common origin, the transition from SBT to LGSC remains poorly understood4. Here, we integrate spatial proteomics5 with spatial transcriptomics to elucidate the evolution from SBT to LGSC and its corresponding metastasis at the molecular level in both the stroma and the tumor. We show that the transition of SBT to LGSC occurs in the epithelial compartment through an intermediary stage with micropapillary features (SBT-MP), which involves a gradual increase in MAPK signaling. A distinct subset of proteins and transcripts was associated with the transition to invasive tumor growth, including the neuronal splicing factor NOVA2, which was limited to expression in LGSC and its corresponding metastasis. An integrative pathway analysis exposed aberrant molecular signaling of tumor cells supported by alterations in angiogenesis and inflammation in the tumor microenvironment. Integration of spatial transcriptomics and proteomics followed by knockdown of the most altered genes or pharmaceutical inhibition of the most relevant targets confirmed their functional significance in regulating key features of invasiveness. Combining cell-type resolved spatial proteomics and transcriptomics allowed us to elucidate the sequence of tumorigenesis from SBT to LGSC. The approach presented here is a blueprint to systematically elucidate mechanisms of tumorigenesis and find novel treatment strategies.

Spatial single-cell mass spectrometry defines zonation of the hepatocyte proteome.

Single-cell proteomics by mass spectrometry is emerging as a powerful and unbiased method for the characterization of biological heterogeneity. So far, it has been limited to cultured cells, whereas an expansion of the method to complex tissues would greatly enhance biological insights. Here we describe single-cell Deep Visual Proteomics (scDVP), a technology that integrates high-content imaging, laser microdissection and multiplexed mass spectrometry. scDVP resolves the context-dependent, spatial proteome of murine hepatocytes at a current depth of 1,700 proteins from a cell slice. Half of the proteome was differentially regulated in a spatial manner, with protein levels changing dramatically in proximity to the central vein. We applied machine learning to proteome classes and images, which subsequently inferred the spatial proteome from imaging data alone. scDVP is applicable to healthy and diseased tissues and complements other spatial proteomics and spatial omics technologies.

Organic Compounds Responsible for the Fouling of Ultrafiltration Membrane Treating Algae-Laden Water.

Fouling comparisons of the organic fractions in surface and algae-laden waters make it possible to determine the main compounds responsible for the fouling of ultrafiltration (UF) membranes. This study examined the fouling of UF membranes and its relationship to the characteristics of the organic fractions found in drinking-water supply. Four types of water were prepared by combining natural organic matter (NOM) from lake water with algal organic matter (AOM) from four algae species commonly found in freshwater. Liquid chromatography-organic carbon detection (LC-OCD) and a fluorescence excitation-emission matrix (FEEM) were used to analyze the feed water and permeate to assess the interactions between and fouling behavior of the organic fractions. The results showed that the interaction of large-molecular-weight AOMs on the membrane surfaces and their transport through the membrane pores were the main fouling mechanisms. Polysaccharides followed by protein-like substances were the organic compounds responsible for the fouling of the UF membranes. The fouling affinity of these substances was attributed to two processes, the adsorption of their carboxyl, hydroxyl and cationic groups on the membrane surfaces, and the molecular complexation of their organic groups. The humic substances' retention was marginal and attributed to the synergetic effects of the polysaccharides and proteins.

A Standardized and Reproducible Workflow for Membrane Glass Slides in Routine Histology and Spatial Proteomics.

Defining the molecular phenotype of single cells in situ is key for understanding tissue architecture in health and disease. Advanced imaging platforms have recently been joined by spatial omics technologies, promising unparalleled insights into the molecular landscape of biological samples. Furthermore, high-precision laser microdissection (LMD) of tissue on membrane glass slides is a powerful method for spatial omics technologies and single-cell type spatial proteomics in particular. However, current histology protocols have not been compatible with glass membrane slides and LMD for automated staining platforms and routine histology procedures. This has prevented the combination of advanced staining procedures with LMD. In this study, we describe a novel method for handling glass membrane slides that enables automated eight-color multiplexed immunofluorescence staining and high-quality imaging followed by precise laser-guided extraction of single cells. The key advance is the glycerol-based modification of heat-induced epitope retrieval protocols, termed "G-HIER." We find that this altered antigen-retrieval solution prevents membrane distortion. Importantly, G-HIER is fully compatible with current antigen retrieval workflows and mass spectrometry-based proteomics and does not affect proteome depth or quality. To demonstrate the versatility of G-HIER for spatial proteomics, we apply the recently introduced deep visual proteomics technology to perform single-cell type analysis of adjacent suprabasal and basal keratinocytes of human skin. G-HIER overcomes previous incompatibility of standard and advanced staining protocols with membrane glass slides and enables robust integration with routine histology procedures, high-throughput multiplexed imaging, and sophisticated downstream spatial omics technologies.

Effects of Oral Cannabidiol on Health and Fitness in Healthy Adults: An 8-Week Randomized Trial.

BACKGROUND: There is a lack of research on the effects of cannabidiol (CBD) on health-related fitness, physical activity, cognitive health, psychological wellbeing, and concentrations of C-reactive protein (CRP) in healthy individuals. CBD has potential anti-inflammatory and neuroprotective effects. METHODS: This study aimed to investigate the effects of 8 weeks of CBD on the above-mentioned measures in healthy individuals. Forty-eight participants were randomized into two groups receiving either oral capsules of 50 mg of CBD or a calorie-matched placebo daily. Participants completed pre- and post-intervention assessments, including blood draws, body composition, fitness, physical activity, and self-reported surveys. RESULTS: There were no significant differences between groups regarding body composition, aerobic fitness, muscular strength, physical activity, cognitive health, psychological wellbeing, and resting CRP concentrations. However, the placebo group experienced a decline in mean peak power and relative peak power compared to the CBD group. CONCLUSIONS: The results suggest that 8 weeks of CBD supplementation may prevent declines in anaerobic fitness over time. However, long-term CBD supplementation may not be beneficial for altering measures of health-related fitness, mental health, and inflammation in healthy individuals.

Association Between Mortality Due to Nasopharyngeal Carcinoma and Race in the United States From 2007 to 2016.

BACKGROUND: Asians and Pacific Islanders (API) exhibit increased incidence of nasopharyngeal carcinoma (NPC). However, they are often excluded when the disease is studied. Risk-factors and incidence are well-researched while cancer-specific mortality trends remain unclear. We aimed to determine whether insurance status modifies the association between race and cancer-specific mortality in NPC patients. METHODS: This retrospective cohort study used secondary data analysis from the Surveillance, Epidemiology, and End Results Program database. Patients ≥18 years with histologically confirmed primary NPC from 2007 - 2016 were included. The main outcome assessed was 5-year survival and the main exposure variable was race (API, white, black). Insurance status was classified into uninsured, any Medicaid, and insured (with any insurance). Potential confounders included age, sex, marital status, stage at diagnosis, and surgical treatment. Adjusted Cox regression analysis was used to calculate hazard ratios (HR) and corresponding 95% confidence intervals (CI). RESULTS: 1610 patients were included (72.98% male, 27.02% female). 49.8% were API, 40.5% were Whites, and 9.8% Blacks. Maximum follow-up was 5-years. The adjusted hazards of 5-year cancer-specific death for API and Blacks compared with Whites were 0.77 (95% CI 0.62 - 0.96) and 0.92 (95% CI 0.65 - 1.31), respectively. Cases decreased with age in API and Blacks. 8.2% of cases had localized disease, 45.3% had local spread, and 44.6% had distant metastasis. Insurance status did not modify the association between race and mortality. CONCLUSION: Race is an important prognostic factor to account for in NPC patients. Investigating risk-factors and subtypes stratified by race may explain our findings.

Structure-Activity Relationship of N-Ethyl-Hexedrone Analogues: Role of the α-Carbon Side-Chain Length in the Mechanism of Action, Cytotoxicity, and Behavioral Effects in Mice.

Synthetic cathinones are ß-keto amphetamine derivatives whose appearance has increased dramatically in the past decades. N-Ethyl substituted cathinones have been proven to potently inhibit dopamine (DA) uptake and induce psychostimulant and rewarding effects in mice. However, little is known about the influence of the alpha-carbon side-chain length of N-ethyl cathinones on their pharmacological and toxicological effects. Thus, the aim of this study was to synthesize and investigate the in vitro and in vivo effects of five N-ethyl substituted cathinones: N-ethyl-cathinone (NEC), N-ethyl-buphedrone (NEB), N-ethyl-pentedrone, N-ethyl-hexedrone (NEH), and N-ethyl-heptedrone. HEK293 cells expressing the human DA or serotonin transporter (hDAT and hSERT) were used for uptake inhibition and binding assays. PC12 cells were used for the cytotoxicity assays. Swiss CD-1 mice were used to study the in vivo psychostimulant, anxiogenic, and rewarding properties. Our results show that all tested cathinones are able to inhibit DA uptake and are DAT-selective. The potency of DA uptake inhibitors increases with the elongation of the aliphatic side chain from methyl to propyl and decreases when increasing from butyl to pentyl, which correlates with an inverted U-shape psychostimulant response in mice at the medium dose tested. On the other hand, an increase in the α-carbon side-chain length correlates with an increase in the cytotoxic properties in PC12 cells, probably due to better membrane penetration. Moreover, all the cathinones tested have shown higher cytotoxicity than methamphetamine. Finally, our study not only demonstrated the rewarding properties of NEC and NEB but also the anxiety-like behavior induced at high doses by all the cathinones tested.

A comparison of resource use of insured and uninsured venezuelan migrants: evidence from the hospital setting.

BACKGROUND: There is no characterization of resource use in the hospital setting for immigrants in Colombia, we aimed to describe the resource use by Venezuelan immigrants, comparing those enrolled in the national health insurance system with those with and without the ability to pay. METHODS: Retrospective review in the billing data system of our Hospital from 2011 to 2020. We collected information for 6,837 hospital episodes associated with 1,022 Venezuelan patients, hospital's billing information for all services rendered was extracted. RESULTS: The mean cost per patient event were 4,595 USD for those without the ability to pay, costing 2.37 times more than a legal resident insured. Care in the ICU, inpatient days, surgery, and OB-GYN department consume most resources provided to vulnerable migrants. DISCUSSION: Enrolment in the national health insurance may allow better access to health services by vulnerable Venezuelan migrants and thus reduce resource use for the health system.

Progression From Patterns A to B in Wellens Syndrome: Electrocardiographic Progression in Wellens Syndrome.

We describe a clinical case series of 3 patients whose electrocardiogram evolved from type A Wellens syndrome to a type B. We emphasize that the diagnosis and treatment for both patterns is the same and that these findings suggest the evolution of the same disease.

The potential of PGPR and Trichoderma-based bioproducts and resistant cultivars as tools to manage clubroot disease in cruciferous crops.

The objective of this research was to determine the potential use of eco-friendly technologies to reduce the clubroot disease caused by Plasmodiophora brassicae, the main constraint of cruciferous crops worldwide. Two commercial bioproducts were evaluated in susceptible broccoli, one based on the PGPR consortium (Bacillus amyloliquefaciens, Bacillus pumilus, and Agrobacterium radiobacter K84) and the other one based on Trichoderma koningiopsis Th003 (Tricotec® WG). Additionally, the resistant broccoli cv. Monclano® was tested under two concentrations of resting spores (RS) of P. brassicae, 1 × 103 and 1 × 105 RS g-1 of soil. The first phase of evaluations with broccoli was carried out under a greenhouse, while susceptible broccoli, cauliflower, and red cabbage were included in a subsequent field phase. Tebuconazole + Trifloxystrobin mixture and Fluazinam were included as positive controls. The effectiveness of the bioproducts depended on the nature of the biocontrol agent, the concentration of P. brassicae, and the dose of treatment. Tricotec® showed consistent plant growth promotion but no biocontrol effect against clubroot, and the rhizobacteria-based bioproduct significantly reduced the disease in both greenhouse and field experiments. Higher disease severity was observed with the higher dose of Tricotec®. Under field conditions, the rhizobacteria reduced the incidence progress by 26%, 39%, and 57% under high, medium, and low pressure of the pathogen, respectively. However, no reduction of clubroot severity under high pressure of the pathogen was observed. Complete inhibition of club formation in roots was achieved via the fungicide, but a phytotoxic effect was observed under greenhouse conditions. Fungicides reduced the incidence progress of clubroot, but not the severity under high inoculum pressure in the field. The fungicides, the bacterial treatment, and the combination of bioproducts tended to delay the progress of the disease compared with the negative control and Tricotec alone. The resistant broccoli showed a low level of disease under high concentrations of P. brassicae (less than 10% incidence and up to 2% severity). These results suggested the overall potential of commercial tools based on the PGPR consortium and plant resistance to control P. brassicae. The integration of control measures, the role of Trichoderma spp. in P. brassicae-cruciferous pathosystems, and the need to recover highly infested soils will be discussed.

Spontaneous Pulmonary Embolism Leading to Sudden Cardiac Arrest and Perimortem C-Section in a 39-Week Parturient During Induction of Labor: A Case Report.

We report the successful salvage of mother and baby after a perimortem cesarean delivery (PMCD) complicated by a 21-minute asystolic maternal cardiac arrest (MCA) that was precipitated by a pulmonary embolism during the early stages of induction of labor. With rapid PMCD, recovery of maternal quality of life is possible even after prolonged resuscitation.

Síndrome antisintetasa: reporte de un caso / Antisynthetase syndrome: a case report

El síndrome antisintetasa es una miopatía inflamatoria idiopática (MII) de origen autoinmune, poco frecuente, que se caracteriza por la presencia de autoanticuerpos antisintetasa ARNt (generalmente anti-Jo1), asociado frecuentemente a miositis, enfermedad pulmonar intersticial, poliartritis, manos de mecánico y fenómeno de Raynaud. Se reporta el caso de una mujer de 45 años de edad que presenta este síndrome con características fenotípicas de dermatomiositis y responde de forma favorable luego de la administración del tratamiento con glucocorticoides asociado a metotrexato.

Anti-synthetase syndrome is a rare autoimmune inflammatory myopathy characterized by autoantibodies against tRNA synthetases (most commonly anti-Jo1) with clinical features that include myositis, interstitial lung disease, polyarthritis, mechanic's hands and Raynaud's phenomenon. We report a 45-year-old woman who presented with dermatomyositis phenotypical features and a significant improvement with corticosteroids and metotrexate treatment.

Síndrome antisintetasa: reporte de un caso / Antisynthetase syndrome: a case repor

El síndrome antisintetasa es una miopatía inflamatoria idiopática (MII) de origen autoinmune, poco frecuente, que se caracteriza por la presencia de autoanticuerpos antisintetasa ARNt (generalmente anti-Jo1), asociado frecuentemente a miositis, enfermedad pulmonar intersticial, poliartritis, manos de mecánico y fenómeno de Raynaud. Se reporta el caso de una mujer de 45 años de edad que presenta este síndrome con características fenotípicas de dermatomiositis y responde de forma favorable luego de la administración del tratamiento con glucocorticoides asociado a metotrexato.

Anti-synthetase syndrome is a rare autoimmune inflammatory myopathy characterized by autoantibodies against tRNA synthetases (most commonly anti-Jo1) with clinical features that include myositis, interstitial lung disease, polyarthritis, mechanic's hands and Raynaud's phenomenon. We report a 45-year-old woman who presented with dermatomyositis phenotypical features and a significant improvement with corticosteroids and metotrexate treatment.

Decreases in resource use after implementing an interdisciplinary management program for patients with placenta accreta spectrum.

OBJECTIVE: Placenta accreta spectrum (PAS) often causes severe morbidity and demands the availability of abundant health resources. Research has shown that the participation of experienced interdisciplinary groups in specialized centers improves clinical outcomes. Our objective is to measure resource use variation after implementing an interdisciplinary management program for this condition. METHODS: Using detailed billing information, hospital care resource use was measured at constant prices for women with PAS who were treated between 2011 and 2019. Cases were classified before (Group 1) and after (Group 2) the implementation of the program. A third group included women with intraoperative MAP findings (Group 3). Comparisons were made using descriptive statistics. RESULTS: The mean reduction in resource use after the program was 16.5% per patient. The program also reduced variability in resource use as measured by the standard deviation and the coefficient of variation, which decreased by 55.2% and 46.3%, respectively. CONCLUSION: The interdisciplinary management of patients affected by PAS in experienced hospitals is associated with a reduction in resource use and variability.

Assessment of Inoculation Methods of Thielaviopsis paradoxa (De Seynes) Höhn into Oil Palm Seedlings under Greenhouse Conditions.

Oil palm (Elaeis guineensis Jacq. and Elaeis Oleifera Cortes) is one of the most important oil crops in the world. Colombia is the fourth-largest oil palm producer worldwide. However, oil palm diseases are a significant factor affecting yield. Thielaviopsis paradoxa (De Seynes) Höhn is a pathogen that affects young palm trees, causing spear rot. Four disease establishment methods were studied to replicate, in a controlled environment, the symptoms of the disease found in the field. Young palm trees were inoculated with a suspension of endoconidia using either local infiltration, drip, scissor cut, or direct contact with agar blocks bearing mycelia and conidia. The effects of the inoculation methods were studied in dose-method-disease severity experiments conducted in a greenhouse under controlled conditions. All four methods resulted in T. paradoxa infections and the development of symptoms of the disease. The disease severity was correlated with the method and dose of inoculation. In trials to test Koch's postulates, T. paradoxa was isolated from areas of disease progression in the inoculated trees, but the teleomorph Ceratocystis paradoxa (Dade) Moreau was not observed. A photographic record of the infection process at different times post-infection was compiled. Given that establishing the disease through artificial inoculation is essential for assessing plant pathogenesis, this study determined that the local infiltration method (1 × 106 endoconidia mL-1) and a 3-7 day incubation period were critical for the development of symptoms as severe as those observed in natural infections in the field.

Multiple de novo gene variations in a progeroid phenotype case report: haploinsufficiency mechanisms.

We are presenting the case of a 6-year-old male patient with progeroid phenotype and severe developmental delay referred to Genetic clinic. Given the complex phenotype an extensive metabolic and genetic evaluation was performed including a whole exome sequencing analysis that showed genetic variants in TTR, RELN, MYH6, PHIP, and SYNE2 genes. Patients' mother and brother were analyzed for the genetic variants in MYH6, PHIP and RELN. Both had same variants on PHIP and RELN as our patient, with no apparent phenotypical consequences. Physical examination was remarkable for dysmorphism including plagiocephaly, low set and abnormally shaped ears, up slanted palpebral fissures, hypoplastic alae nasi, and a head circumference two standard deviations below the 3rd percentile (microcephaly). Other characteristics include wrinkled skin, a broad forehead, sparse eyelashes in lower eyelid, short palpebral fissures, upturned nares, thick lips, right occipital plagiocephaly, overfolded helix and prominent anti-helix, protuberant chest, scaphoid abdomen, digitalized thumbs, and kyphosis due to low muscle tone. The patient presented abnormal EEG with evidence of epileptic discharges. A temporal bone CT showed plagiocephaly with flattening of the right occipital bone. Brain MRI showed callosal agenesis with bilateral colpocephaly with temporal horn dilatation, parahippocampal atrophy, lissencephaly and midbrain hypoplasia. The combination of de novo gene variants mentioned above has never been reported nor correlated as the result of haploinsufficiency mechanisms. Thus, we propose haploinsufficiency and loss of heterozygosity as etiological reasons for this patient phenotype. Further proteomic studies are needed to allocate the extense of genetic influence within the clinical manifestations.

Post-severe Acute Respiratory Syndrome Coronavirus 2 Monoclonal Antibody Treatment Hospitalizations as a Sentinel for Emergence of Viral Variants in New York City.

We partnered with the US Department of Health and Human Services to treat high-risk, nonadmitted coronavirus disease 2019 (COVID-19) patients with bamlanivimab in the Bronx, New York per Emergency Use Authorization criteria. Increasing posttreatment hospitalizations were observed monthly between December 2020 and March 2021 in parallel to the emergence of severe acute respiratory syndrome coronavirus 2 variants in New York City.

Caso Clínico: Lipoblastoma en un Lactante / Clinical Case: Lipoblastoma in an Infant

Introducción: El lipoblastoma es una neoplasia benigna de presentación infrecuente originada en el tejido adiposo, de presentación casi exclusiva en pacientes pediátricos antes de los tres años de edad, con predominio en el sexo masculino. Se localiza principalmente en las extremidades y tronco, como una tumoración indolora de crecimiento progresivo. El tratamiento de elección es quirúrgico y tiene pronóstico favorable. Las recidivas se presentan en los casos en los que la resección no pudo ser completa. Caso clínico: Lactante mujer de 7 meses de edad. Desde los tres meses de edadpresentó aumento progresivo del volumen de la extremidad inferior izquierda.Al examen físico se evidenciógran tumoraciónde 12 x 7 centímetrosque compromete la cara posterior del muslo izquierdo: Masa indolora y bien delimitada. Sin compromiso de la movilidad, sin edema de miembro. Taller diagnóstico: La Resonancia Magnética reportó unatumoración sólida de muslo izquierdo hiper-intensa en T1 y T2, hipo-intensa en STIR con septos finos en su interior, se extiende desde el musculo abductor magno del bíceps femoral y mide 11.2 x 7.9 x 8.4 cm en sus ejes longitudinal, anteroposterior y transversal respectivamente, desplazando y comprimiendo a los músculos semitendinoso, semimembranoso y gracilis. La paciente fue sometida a escisión completa y amplia de la masa, mediante abordaje posterolateral. Desenlace:Patología reportóun tumor constituido por tejido adiposo con tejido mixoide,sin atipia nuclear; se evidencia infiltración parcial del musculo esquelético adyacente, sin afectación de piel, sin necrosisy bordes quirúrgicos negativos para neoplasia. Inmunohistoquímica con marcador de proliferación celular KI-67 resultado positivo de 1%, y estudio de MDM2 (inhibidor de la activación transcripcional de p53) negativo; hallazgos compatibles con lipoblastoma. El estudio citogenético no fue realizado. Evolución: La paciente fue dada de alta al cuarto día post-operatorio sin complicaciones. Con una recuperación completa, en el quinto mes de seguimiento se realizónuevo estudio de resonancia magnética en la cual no se visualizan imágenes que sugieran tumor residual ni recidiva tumoral. Conclusión: El lipoblastoma debe ser tomada en cuenta como diagnóstico diferencial en niños con tumores de partes blandas, su tratamiento es eminentemente quirúrgico con un buen pronóstico si la extirpación es completa

Introduction: Lipoblastoma is an infrequent benign neoplasm originating in adipose tissue, presenting almost exclusively in pediatric patients before the age of three years, predominantly in males. It is located mainly on the limbs and trunk, as a painless, progressively growing tumor. The treatment of choice is surgical and has a favorable prognosis. Recurrences occur in cases in which the resection could not be complete. Clinical case: 7-month-old female infant. From the age of three months, he presented a progressive increase in the volume of the left lower limb. Physical examination revealed a large mass measuring 12 x 7 centimeters that involves the posterior aspect of the left thigh: a painless and well-defined mass. No compromise of mobility, no limb edema. Diagnostic workshop: Magnetic resonance imaging reported a hyper-intense solid tumor of the left thigh in T1 and T2, hypo-intense in STIR with fine septa inside it, extending from the abductor magnus muscle of the biceps femoris and measuring 11.2 x 7.9 x 8.4 cm in its longitudinal, anteroposterior and transverse axes respectively, displacing and compressing the semitendinosus, semimembranosus and gracilis muscles. The patient underwent a complete and wide excision of the mass using a posterolateral approach. Outcome: Pathology reported a tumor made up of adipose tissue with myxoid tissue, without nuclear atypia; partial infiltration of the adjacent skeletal muscle is evidenced, without skin involvement, without necrosis and negative surgical margins for neoplasia. Immunohistochemistry with a cell proliferation marker KI-67, a positive result of 1%, and a negative MDM2 study (inhibitor of transcriptional activation of p53); findings consistent with lipoblastoma. The cytogenetic study was not performed. Evolution:The patient was discharged on the fourth postoperative day without complications. With a complete recovery, in the fifth month of follow-up a new magnetic resonance study was performed in which no images that suggest residual tumor or tumor recurrence are visualized Conclusion: Lipoblastoma should be taken into account as a differential diagnosis in children with soft tissue tumors, its treatment is eminently surgicalwith a good prognosis if the removal is complete

Putative Novel Effector Genes Revealed by the Genomic Analysis of the Phytopathogenic Fungus Fusarium oxysporum f. sp. physali (Foph) That Infects Cape Gooseberry Plants.

The vascular wilt disease caused by the fungus Fusarium oxysporum f. sp. physali (Foph) is one of the most limiting factors for the production and export of cape gooseberry (Physalis peruviana) in Colombia. A transcriptomic analysis of a highly virulent strain of F. oxysporum in cape gooseberry plants, revealed the presence of secreted in the xylem (SIX) effector genes, known to be involved in the pathogenicity of other formae speciales (ff. spp.) of F. oxysporum. This pathogenic strain was classified as a new f. sp. named Foph, due to its specificity for cape gooseberry hosts. Here, we sequenced and assembled the genome of five strains of F. oxysporum from a fungal collection associated to the cape gooseberry crop (including Foph), focusing on the validation of the presence of SIX homologous and on the identification of putative effectors unique to Foph. By comparative and phylogenomic analyses based on single-copy orthologous, we found that Foph is closely related to F. oxysporum ff. spp., associated with solanaceous hosts. We confirmed the presence of highly identical homologous genomic regions between Foph and Fol that contain effector genes and identified six new putative effector genes, specific to Foph pathogenic strains. We also conducted a molecular characterization using this set of putative novel effectors in a panel of 36 additional stains of F. oxysporum including two of the four sequenced strains, from the fungal collection mentioned above. These results suggest the polyphyletic origin of Foph and the putative independent acquisition of new candidate effectors in different clades of related strains. The novel effector candidates identified in this genomic analysis, represent new sources involved in the interaction between Foph and cape gooseberry, that could be implemented to develop appropriate management strategies of the wilt disease caused by Foph in the cape gooseberry crop.