Toward a molecular microbial blood test for tuberculosis infection.

The World Health Organization's aim to end the global tuberculosis (TB) epidemic by 2050 cannot be achieved without taking measures to identify people with asymptomatic Mycobacterium tuberculosis (Mtb) infection and offer them an intervention to reduce the risk of disease progression, such as preventive antimicrobial therapy. Implementation of this strategy is limited by the fact that existing tests for Mtb infection, which use immunosensitization to Mtb-specific antigens as a proxy for infection, have low positive predictive value for progression to TB. A blood test that detects Mtb deoxyribonucleic acid (DNA) could allow preventive therapy to be targeted at individuals with microbiological evidence of persistent infection. In this review, we summarize recent advances in the development of molecular microbial blood tests for Mtb infection and discuss potential explanations for discordance between their results and those of immunodiagnostic tests in adults with recent exposure to an infectious index case. We also present a roadmap for further development of molecular microbial blood tests for Mtb infection, and highlight the potential for research in this area to provide novel insights into the biology of Mtb infection and yield new tools to support efforts to control the global TB epidemic.

First report of Pectobacterium brasiliense causing banana soft rot in Ecuador.

Banana (Musa spp.) is the most economically important crop in Ecuador, with exports representing 35% of the agricultural GDP of the country. It covers 230,000 hectares, mostly concentrated in three coastal provinces, Guayas, Los Ríos, and El Oro. Between July and September 2022, disease symptomatic banana cv. Williams plants were observed in commercial plantations located in two parishes in the province of Guayas (Naranjito and Lorenzo de Garaicoa) and one parish in the province of Santo Domingo de los Tsáchilas (La Concordia), with an incidence that ranged from 5% to 15%. Symptoms included soft rot of the pseudostem and rhizome decay, characterized by a fetid odor. Three symptomatic pseudostems from each location were collected, washed with running water to remove any debris, and dried with absorbent paper. From the lesion of each pseudostem, seven pieces of 2 cm² were taken, surface-sterilized, and macerated in 9 ml of sterile peptone water (0.1% w/v). The macerate was diluted three fold in sterile water, plated on nutrient agar, and incubated at 30°C for 24 h. Eight randomly picked colonies, with convex elevation and creamy white color, were isolated on nutrient agar. Each of the bacterial isolates was biochemically profiled by the Biolog system (Biolog Inc., USA) and identified as Pectobacterium. Three isolates, one from each parish (FP220416, FP220694, and FP220904), were selected for testing Koch's postulates and further identification. Sequences from fragments of the 16S, dnaA, gapA and gyrB genes were obtained from these isolates, following the protocols used by Dobhal et al. (2020) and Boluk et al. (2020), showing 98.1-99.0%, 98.2%, 99.7-99.8%, and 98.4-98.9% identitity, respectively, with sequences from the P. brasiliense type strain LMG_21371 (Acc. number JQOE00000000). The obtained sequences were deposited in GenBank with the following accession numbers: OR392417, OR371545,OR371546, OR727281, OR727282, and OR739074-OR739080. Using BEAST v.1.10.4 (Suchard et al.,2018), a bayesian multilocus phylogenetic tree was built with multiple sequence alignments of dnaA, gapA, ang gyrB from 22 P. brasiliense isolates and 2 P. aquaticum isolates used as outgroup. The phylogenetic analysis showed that the Ecuadorian isolates cluster with P. brasiliense BF20, isolated from Opuntia ficus-indica in México and are closely related with the type strain. Pathogenicity tests were conducted through syringe infiltration with 1 ml of 1 × 10^8 CFU ml-1 bacterial suspensions. Each of the three characterized isolates were inoculated into the pseudostems of five healthy 4-month-old banana plants of the Williams cultivar. Negative control plants were infiltrated with sterile distilled water. The plants were incubated at 25°C and 74% relative humidity. Black lesions started to appear 11 days after inoculation and 5 weeks after inoculation plants showed clear symptoms of soft rot of the pseudostem, including fetid odor associated with plant tissue decomposition. Control plants remained symptom-free. Bacteria were re-isolated only from symptomatic pseudostems and identified as P. brasiliense with specific primers Pb1F and Pb1R. Soft rot of banana caused by different enterobacteria including Dickeya zeae, Erwinia carotovora, and Erwinia chrysanthemi hasve been previously reported (Jingxin et al. 2022, Arun et al. 2012, Loganathan, et al. 2019). This is the first report of P. brasiliense causing soft rot of banana in Ecuador, the biggest exporter of the fruit in the world.

Percepción de las fuentes de apoyo que facilitan la adherencia a los tratamientos / Perception of sources of support that facilitate adherence to the treatments

Introducción: La falta de adherencia a los tratamientos aumenta la probabilidad de fracaso terapéutico y complicaciones innecesarias. Objetivo: Explorar las fuentes de apoyo percibidas por los pacientes con enfermedades crónicas que les facilitan la adherencia a los tratamientos. Método: Estudio cualitativo descriptivo con enfoque fenomenológico, en el que se reclutaron ocho pacientes crónicos complejos, sin distinción de género, ni edad, residentes en Barcelona, y que tras evaluar la adherencia farmacológica con los test de Morisky-Green y el test de Batalla diera como resultado ser adherente al régimen terapéutico. Tras lo cual se efectuaron dos preguntas abiertas que fueron grabadas en un dispositivo de audio y transcritas textualmente. Para el análisis de los datos, se utilizó el método Colaizzi. Resultados: Surgieron tres grupos temáticos: 1) Tener el apoyo familiar; 2) Comprensión de la enfermedad y estrategias que facilitan la adhesión a los tratamientos; y 3) Relación con los profesionales de referencia. Conclusiones: La fuente más importante de apoyo percibida por los participantes, es la familia, coincidiendo con otros autores. El hallazgo de este estudio, es que se evidencia a medicina y enfermería como instigadores que fomentan su empoderamiento y como fuente de influencia reconocida que ha permitido su asimilación y capacitación para adoptar medidas que les ayudan a seguir las pautas farmacoterapéuticas. (AU)

Introduction: Non-adherence to treatments increases the probability of therapeutic failure and unnecessary complications. Objective: To explore the sources of support perceived by patients with chronic diseases that facilitate adherence to treatment. Method: Qualitative descriptive study with a phenomenological approach, in which eight complex chronic patients were recruited, without distinction of gender or age, residents of Barcelona, and who, after evaluating pharmacological adherence with the Morisky-Green test and the test of Batalla resulted in being adherent to the therapeutic regimen.After which, two open questions were asked that were recorded on an audio device and transcribed verbatim. For data analysis, the method Colaizzi was used. Results: Three thematic groups emerged: 1) Having family support; 2) Understanding of the disease and strategies that facilitate adherence to treatments; and 3) Relationship with reference professionals. Conclusions: The most important source of support perceived by the participants is the family, coinciding with other authors. The finding of this study is that shows medicine and nursing are evidenced as instigators that promote their empowerment and as a recognized source of influence that has allowed their assimilation and training to adopt measures that help them follow the pharmacotherapeutic guidelines. (AU)

Correction: The Meaning of Lymphadenopathies During Adjuvant Durvalumab After Chemoradiotherapy for Lung Cancer: Thinking Beyond Disease Progression.

[This corrects the article DOI: 10.7759/cureus.26729.].

MicroRNA-27a-3p targets FoxO signalling to induce tumour-like phenotypes in bile duct cells.

BACKGROUND & AIMS: Cholangiocarcinoma (CCA) is a heterogeneous and lethal malignancy, the molecular origins of which remain poorly understood. MicroRNAs (miRs) target diverse signalling pathways, functioning as potent epigenetic regulators of transcriptional output. We aimed to characterise miRNome dysregulation in CCA, including its impact on transcriptome homeostasis and cell behaviour. METHODS: Small RNA sequencing was performed on 119 resected CCAs, 63 surrounding liver tissues, and 22 normal livers. High-throughput miR mimic screens were performed in three primary human cholangiocyte cultures. Integration of patient transcriptomes and miRseq together with miR screening data identified an oncogenic miR for characterization. MiR-mRNA interactions were investigated by a luciferase assay. MiR-CRISPR knockout cells were generated and phenotypically characterized in vitro (proliferation, migration, colony, mitochondrial function, glycolysis) and in vivo using subcutaneous xenografts. RESULTS: In total, 13% (140/1,049) of detected miRs were differentially expressed between CCA and surrounding liver tissues, including 135 that were upregulated in tumours. CCA tissues were characterised by higher miRNome heterogeneity and miR biogenesis pathway expression. Unsupervised hierarchical clustering of tumour miRNomes identified three subgroups, including distal CCA-enriched and IDH1 mutant-enriched subgroups. High-throughput screening of miR mimics uncovered 71 miRs that consistently increased proliferation of three primary cholangiocyte models and were upregulated in CCA tissues regardless of anatomical location, among which only miR-27a-3p had consistently increased expression and activity in several cohorts. FoxO signalling was predominantly downregulated by miR-27a-3p in CCA, partially through targeting of FOXO1. MiR-27a knockout increased FOXO1 levels in vitro and in vivo, impeding tumour behaviour and growth. CONCLUSIONS: The miRNomes of CCA tissues are highly remodelled, impacting transcriptome homeostasis in part through regulation of transcription factors like FOXO1. MiR-27a-3p arises as an oncogenic vulnerability in CCA. IMPACT AND IMPLICATIONS: Cholangiocarcinogenesis entails extensive cellular reprogramming driven by genetic and non-genetic alterations, but the functional roles of these non-genetic events remain poorly understood. By unveiling global miRNA upregulation in patient tumours and their functional ability to increase proliferation of cholangiocytes, these small non-coding RNAs are implicated as critical non-genetic alterations promoting biliary tumour initiation. These findings identify possible mechanisms for transcriptome rewiring during transformation, with potential implications for patient stratification.

Identification and Characterization of Pythium, Globisporangium, and Phytopythium Species Present in Floricultural Crops from Long Island, New York.

Several Pythium, Globisporangium, and Phytopythium species cause Pythium diseases in greenhouse floricultural crops, resulting in significant seasonal losses. Four hundred and eighteen Pythium, Globisporangium, and Phytopythium isolates from flowering crops, growing media, or bench and floor debris were collected from Long Island greenhouses or clinic samples between 2002 and 2013. Isolates were identified to species based on morphology and internal transcribed spacer barcoding. Twenty-two species of Pythium, Phytopythium, and Globisporangium were identified, with Globisporangium irregulare sensu lato (s.l.) being the most common. To determine the origin of inoculum during the 2011 cropping season, 11 microsatellite loci were analyzed in 124 G. irregulare s.l. isolates collected in four greenhouses and six previously collected from clinic samples. Cluster analyses grouped G. irregulare s.l. isolates into four groups: G. irregulare sensu stricto, plus three G. cryptoirregulare clusters. The population structure defined by greenhouse and host was found in two clades. Additionally, the population dynamics of G. irregulare s.l. isolates associated with Pelargonium spp. from 2011 to 2013 were examined using 85 isolates and nine informative microsatellite loci to assess inoculum survival over multiple cropping seasons. Although most isolates had unique genotypes, closely related genotypes were found in the same locations over different years. Our results indicate that G. irregulare s.l. inocula have local as well as remote origins. Isolates may be initially brought into ornamental operations from common sources, such as infected plant materials or infested potting mixes. Our results support the hypothesis that established strains can serve as inocula and survive in greenhouse facilities over multiple seasons.

The Meaning of Lymphadenopathies During Adjuvant Durvalumab After Chemoradiotherapy for Lung Cancer: Thinking Beyond Disease Progression.

Immune-checkpoint inhibitors (ICIs) have become the mainstay of treatment for many malignancies. With this new strategy, relevant immune-related adverse events (irAEs) have been reported, some of which can be mistaken for disease progression. To better illustrate the current challenges in diagnosing and managing a patient under adjuvant ICI treatment, we present the case of a 67-year-old female patient with stage IIIB unresectable, epidermal growth factor receptor (EGFR)-mutated, non-small-cell lung cancer who was initially treated with chemoradiotherapy, followed by immunotherapy with durvalumab. During the course of immunotherapy, the patient presented with madarosis and erythematous and endured skin lesions, in addition to lymphadenopathies and pulmonary infiltrates. She was started on first-line palliative treatment with an EGFR tyrosine kinase inhibitor. After reviewing the case, a multidisciplinary team meeting suggested diagnostic procedures, including a transbronchial needle aspiration from mediastinal lymph nodes. The histologic examination showed chronic systemic inflammation and non-caseating granulomas of the sarcoid type. In this case, palliative treatment was suspended and systemic therapy with prednisolone was initiated. The patient became asymptomatic and the previously observed radiologic abnormalities resolved. This case highlights the importance of early recognition and appropriate treatment of irAEs, mainly because these conditions remain poorly understood and are probably underdiagnosed. Considering differential diagnosis is paramount to guide clinical management, despite curative or palliative treatment intent.

Whole blood microRNAs capture systemic reprogramming and have diagnostic potential in patients with biliary tract cancer.

BACKGROUND & AIMS: Late diagnosis is a critical factor undermining clinical management of patients with biliary tract cancer (BTC). While biliary tumours display extensive inter-patient heterogeneity, the host immune response may be comparatively homogenous, providing diagnostic opportunities. Herein, we investigated whether cancer-associated systemic reprogramming could be detected non-invasively to improve diagnosis of BTC. METHODS: In this prospective Danish study, whole blood (WB) microRNA (miRNA) profiling was performed in samples from 218 patients with BTC, 99 healthy participants, and 69 patients with differential diagnoses split into discovery (small RNA-sequencing) and validation (RT-qPCR) cohorts. miRNA expression and activity were further investigated in 119 and 660 BTC tissues, respectively. RESULTS: Four WB miRNAs (let-7a-3p, miR-92b-5p, miR-145-3p, miR-582-3p) were identified and validated as diagnostic of BTC on univariable analysis. Two diagnostic miRNA indexes were subsequently identified that were elevated in patients with BTC and in patients with differential diagnoses, compared to healthy participants. The combination of these miRNA indexes with serum CA 19-9 significantly improved the diagnostic performance of CA 19-9 alone, consistently achieving superior AUC values irrespective of clinical setting (minimum AUC >0.84) or tumour location (minimum AUC >0.87). The diagnostic information captured by miRNA indexes was not recapitulated by routine clinical measurements. Index miRNA expression in BTC tissues was associated with distinct pathobiological and immune features. CONCLUSIONS: WB miRNA profiles are altered in patients with BTC. Quantification of miRNA indexes in combination with serum CA 19-9 has the potential to improve early diagnosis of BTC, pending further validation. LAY SUMMARY: Surgery is currently the only curative intervention for patients with biliary tract cancer (BTC). However, resection is not possible for most patients who are diagnosed with late-stage disease. With the aim of identifying new early diagnostic opportunities, we analysed circulating microRNAs (small non-coding RNAs whose role in cancer is being increasingly recognised) in whole blood samples. We identified a microRNA signature that could distinguish patients with BTC from healthy participants. These miRNAs significantly improved the diagnostic potential of the routinely measured biomarker, CA 19-9, and were implicated in distinct immune processes in tumour tissues.

The protease-inhibitor SerpinB3 as a critical modulator of the stem-like subset in human cholangiocarcinoma.

BACKGROUND AND AIMS: Cholangiocarcinoma (CCA) is a form of primary liver cancer with limited therapeutic options. Recently, cancer stem cells (CSCs) have been proposed as a driving force of tumour initiation and dissemination, thus representing a crucial therapeutic target. The protease inhibitor SerpinB3 (SB3) has been identified in several malignancies including hepatocellular carcinoma. SB3 has been involved in the early events of hepatocarcinogenesis and is highly expressed in hepatic progenitor cells and in a mouse model of liver progenitor cell activation. However, only limited information on the possible role of SB3 in CCA stem-like compartment is available. METHODS: Enrichment of CCA stem-like subset was performed by sphere culture (SPH) in CCA cell lines (CCLP1, HUCCT1, MTCHC01 and SG231). Quantitative RT-PCR and Western blotting were used to detect SB3 in both SPH and parental monolayer (MON) cells. Acquired CSC-like features were analysed using an endogenous and a paracrine in vitro model, with transfection of SB3 gene or addition of recombinant SB3 to cell medium respectively. SB3 tumorigenic role was explored in an in vivo mouse model of CCA by subcutaneous injection of SB3-transfected MON (MONSB3+ ) cells in immune-deficient NOD-SCID/IL2Rgnull  (NSG) mice. SB3 expression in human CCA sections was investigated by immunohistochemistry. Overall survival (OS) and time to recurrence (TTR) analyses were carried out from a transcriptome database of 104 CCA patients. RESULTS: SB3, barely detected in parental MON cells, was overexpressed in the same CCA cells grown as 3D SPH. Notably, MONSB3+ showed significant overexpression of genes associated with stemness (CD24, CD44, CD133), pluripotency (c-MYC, NOTCH1, STAT3, YAP, NANOG, BMI1, KLF4, OCT4, SOX2), epithelial mesenchymal transition (ß-catenin, SLUG) and extracellular matrix remodelling (MMP1, MMP7, MMP9, ADAM9, ADAM10, ADAM17, ITGB3). SB3-overexpressing cells showed superior spherogenic capacity and invasion ability compared to control. Importantly, MONSB3+ exhibited activation of MAP kinases (ERK1/2, p38, JNK) as well as phosphorylation of NFκB (p65) in addition to up-regulation of the proto-oncogene ß-catenin. All these effects were reversed after transient silencing of SB3. According to the in vitro finding, MONSB3+ cells retained high tumorigenic potential in NSG mice. SB3 overexpression was observed in human CCA tissues and analysis of OS as well as TTR indicated a worse prognosis in SB3+ CCA patients. CONCLUSION: These findings indicate a SB3 role in mediating malignant phenotype of CCA and identify a new therapeutic target.

Linfangiomatosis pulmonar difusa con compromiso pleural y pericárdico. Reporte de un caso pediátrico / Diffuse pulmonary lymphangiomatosis with pleural and pericardial involvement. Pediatric case report

La linfangiomatosis pulmonar difusa es una enfermedad rara caracterizada por una marcada proliferación y dilatación de los vasos linfáticos en los pulmones, la pleura y el mediastino. Se desconoce la prevalencia, y la etiología no se comprende completamente.Una niña de 22 meses ingresó por poliserositis, con derrame pericárdico y pleural. Requirió pericardiocentesis y avenamiento pleural, y presentó drenaje de quilo (1,5-4 litros/día) sin respuesta al tratamiento médico (ayuno, nutrición parenteral y octreotide). Se realizó biopsia pulmonar. La anatomía patológica mostró hallazgos compatibles con linfangiomatosis difusa pulmonar. Comenzó tratamiento con sirolimus y propanolol, que disminuyeron las pérdidas por el drenaje pleural a la semana. Presentó buena evolución; suspendió aporte de oxígeno y se retiró el drenaje pleural. Se externó al cuarto mes de internación. El diagnóstico temprano de la linfangiomatosis pulmonar difusa es difícil de lograr, pero permite aplicar terapéuticas que evitan la progresión de enfermedad y disminuir la morbimortalida

Diffuse pulmonary lymphangiomatosis is a rare disease characterized by marked proliferation and dilation of lymphatic vessels in the lungs, pleura, and mediastinum. The prevalence is unknown and the etiology is not fully understood.A 22-month-old girl was admitted for polyserositis, with pericardial and pleural effusion. She required pericardiocentesis and pleural drainage, presenting chyle drainage (1.5-4 liters/day) without response to medical treatment (fasting, parenteral nutrition and octreotide). A lung biopsy was performed. The pathological anatomy showed findings compatible with diffuse pulmonary lymphangiomatosis. Treatment with sirolimus and propanolol began, decreasing losses due to pleural drainage one week after treatment. She progressed well, discontinued oxygen supply and pleural drainage was removed, leaving the patient after the fourth month of hospitalization.Early diagnosis of diffuse pulmonary lymphangiomatosis is difficult to achieve, but it allows the application of therapies that prevent disease progression, reducing morbidity and mortality.

First Report of Pantoea ananatis causing leaf spot disease of maize in Ecuador.

Maize (Zea mays) is the second most cultivated grain crop in Ecuador, with growing significance as a source of fodder and food. During the rainy season (November and December) of 2018 and 2019, a disease of maize that was not previously observed in Ecuador was found at commercial fields of Misqui Sara variety, at four parishes of canton Quito (Tumbaco, Pifo, Puembo, and Checa), province of Pichincha. Infected plants, at tassel initiation, displayed symptoms of localized chlorotic streaks on leaves that expanded with time, and around a month later turned necrotic. Severely affected plants wilted and died. Symptoms appeared in lower leaves first and were later observed in upper leaves as the disease progressed. Disease incidence was between 20 and 30% in the affected plantations, with around 30% of infected plants wilting and dying, resulting in 20-25% of yield losses. Upper leaves from ten symptomatic plants, five from Puembo and five from Checa, were collected randomly. Two 0.5 cm2 pieces of leaf from each plant were excised from the margins of the necrotic lesions, surface sterilized and macerated in 9 mL of sterile peptone water. The 10-3 dilutions were plated onto nutrient agar and incubated at 28°C for 24 hours. Yellow, mucoid colonies were isolated on nutrient agar. Three isolates from Puembo and two from Checa were selected for testing Koch´s postulates and further biochemical and molecular characterization. Isolates were Gram-negative rods, oxidase negative, catalase, indol and citrate positive. Fragments of the 16S, gyrB, and rpoB loci were amplified and sequenced using the 27F/1492R (Lane, D. J., 1991), UP-1/UP-2r (Yamamoto & Harayama, 1995), and rpoBCM81-F/rpoBCM32b-R (Brady, C., et al., 2008) primer pairs, respectively. All isolates presented identical sequences for the different loci, therefore only sequences from isolate FP191505 were deposited in GenBank (GenBank accession no. MW528428-MW528430). A search of homologous sequences using BLAST resulted in identities of 99.3, 99.7, and 100 % for 16S, gyrB, and rpoB, respectively, with sequences from Pantoea ananatis type specimen LGM 2665 (Brady, C., et al., 2008; Hauben, L., et al., 1998; GenBank accession nos NR_119362.1, EF988824.1 EF988996.1), indicating that our isolates belong to this species. Pathogenicity tests were performed by syringe infiltration of bacterial suspensions. Each one of the five characterized P. ananatis isolates was inoculated in four leaves (500 ul of 1 x 108 CFU mL-1 per leave) of three healthy maize plants. Negative control plants were infiltrated with sterile distilled water. Plants were incubated at 28-30°C and 60% relative humidity for 24 hours. Later, plants were maintained in a greenhouse with 27°C/21°C day/night temperatures and observed daily. After six weeks all bacteria-inoculated plants developed symptoms of chlorosis and necrosis while the control was symptomless. Bacteria were re-isolated from symptomatic leaves and identified as P. ananatis following the same methodologies used for the initial identification. To our knowledge, this is the first report of P. ananatis causing leaf spot of maize in Ecuador.

[Diffuse pulmonary lymphangiomatosis with pleural and pericardial involvement. Pediatric case report]. / Linfangiomatosis pulmonar difusa con compromiso pleural y pericárdico. Reporte de un caso pediátrico.

Diffuse pulmonary lymphangiomatosis is a rare disease characterized by marked proliferation and dilation of lymphatic vessels in the lungs, pleura, and mediastinum. The prevalence is unknown and the etiology is not fully understood. A 22-month-old girl was admitted for polyserositis, with pericardial and pleural effusion. She required pericardiocentesis and pleural drainage, presenting chyle drainage (1.5-4 liters/ day) without response to medical treatment (fasting, parenteral nutrition and octreotide). A lung biopsy was performed. The pathological anatomy showed findings compatible with diffuse pulmonary lymphangiomatosis. Treatment with sirolimus and propanolol began, decreasing losses due to pleural drainage one week after treatment. She progressed well, discontinued oxygen supply and pleural drainage was removed, leaving the patient after the fourth month of hospitalization. Early diagnosis of diffuse pulmonary lymphangiomatosis is difficult to achieve, but it allows the application of therapies that prevent disease progression, reducing morbidity and mortality.

La linfangiomatosis pulmonar difusa es una enfermedad rara caracterizada por una marcada proliferación y dilatación de los vasos linfáticos en los pulmones, la pleura y el mediastino. Se desconoce la prevalencia, y la etiología no se comprende completamente. Una niña de 22 meses ingresó por poliserositis, con derrame pericárdico y pleural. Requirió pericardiocentesis y avenamiento pleural, y presentó drenaje de quilo (1,5- 4 litros/día) sin respuesta al tratamiento médico (ayuno, nutrición parenteral y octreotide). Se realizó biopsia pulmonar. La anatomía patológica mostró hallazgos compatibles con linfangiomatosis difusa pulmonar. Comenzó tratamiento con sirolimus y propanolol, que disminuyeron las pérdidas por el drenaje pleural a la semana. Presentó buena evolución; suspendió aporte de oxígeno y se retiró el drenaje pleural. Se externó al cuarto mes de internación. El diagnóstico temprano de la linfangiomatosis pulmonar difusa es difícil de lograr, pero permite aplicar terapéuticas que evitan la progresión de enfermedad y disminuir la morbimortalidad.

Molecular Targets in Cholangiocarcinoma.

Cholangiocarcinoma (CCA) encompasses a heterogeneous collection of malignancies for which diagnostic biomarkers are lacking and population screening is infeasible because of its status as a rare disease. Coupled with high postsurgical recurrence rates among the minority of patients diagnosed at resectable stages, systemic clinical management will inevitably be required for the majority of patients with CCA with recurrent and advanced disease. In this review, we discuss the therapeutic potential of different classes of molecular targets at various stages of development in CCA, including those targeted to the tumor epithelia (oncogenic, developmental, metabolic, epigenomic) and tumor microenvironment (angiogenesis, checkpoint regulation). Furthermore, we discuss the successes and failures of CCA-targeted therapies, emphasizing key lessons learned that should pave the way for future molecular target evaluation in this uncommon yet bona fide target-rich disease.

TREM-2 defends the liver against hepatocellular carcinoma through multifactorial protective mechanisms.

OBJECTIVE: Hepatocellular carcinoma (HCC) is a prevalent and aggressive cancer usually arising on a background of chronic liver injury involving inflammatory and hepatic regenerative processes. The triggering receptor expressed on myeloid cells 2 (TREM-2) is predominantly expressed in hepatic non-parenchymal cells and inhibits Toll-like receptor signalling, protecting the liver from various hepatotoxic injuries, yet its role in liver cancer is poorly defined. Here, we investigated the impact of TREM-2 on liver regeneration and hepatocarcinogenesis. DESIGN: TREM-2 expression was analysed in liver tissues of two independent cohorts of patients with HCC and compared with control liver samples. Experimental HCC and liver regeneration models in wild type and Trem-2-/- mice, and in vitro studies with hepatic stellate cells (HSCs) and HCC spheroids were conducted. RESULTS: TREM-2 expression was upregulated in human HCC tissue, in mouse models of liver regeneration and HCC. Trem-2-/- mice developed more liver tumours irrespective of size after diethylnitrosamine (DEN) administration, displayed exacerbated liver damage, inflammation, oxidative stress and hepatocyte proliferation. Administering an antioxidant diet blocked DEN-induced hepatocarcinogenesis in both genotypes. Similarly, Trem-2-/- animals developed more and larger tumours in fibrosis-associated HCC models. Trem-2-/- livers showed increased hepatocyte proliferation and inflammation after partial hepatectomy. Conditioned media from human HSCs overexpressing TREM-2 inhibited human HCC spheroid growth in vitro through attenuated Wnt ligand secretion. CONCLUSION: TREM-2 plays a protective role in hepatocarcinogenesis via different pleiotropic effects, suggesting that TREM-2 agonism should be investigated as it might beneficially impact HCC pathogenesis in a multifactorial manner.

Synthetic Conjugates of Ursodeoxycholic Acid Inhibit Cystogenesis in Experimental Models of Polycystic Liver Disease.

BACKGROUND AND AIMS: Polycystic liver diseases (PLDs) are genetic disorders characterized by progressive development of symptomatic biliary cysts. Current surgical and pharmacological approaches are ineffective, and liver transplantation represents the only curative option. Ursodeoxycholic acid (UDCA) and histone deacetylase 6 inhibitors (HDAC6is) have arisen as promising therapeutic strategies, but with partial benefits. APPROACH AND RESULTS: Here, we tested an approach based on the design, synthesis, and validation of a family of UDCA synthetic conjugates with selective HDAC6i capacity (UDCA-HDAC6i). Four UDCA-HDAC6i conjugates presented selective HDAC6i activity, UDCA-HDAC6i #1 being the most promising candidate. UDCA orientation within the UDCA-HDAC6i structure was determinant for HDAC6i activity and selectivity. Treatment of polycystic rats with UDCA-HDAC6i #1 reduced their hepatomegaly and cystogenesis, increased UDCA concentration, and inhibited HDAC6 activity in liver. In cystic cholangiocytes UDCA-HDAC6i #1 restored primary cilium length and exhibited potent antiproliferative activity. UDCA-HDAC6i #1 was actively transported into cells through BA and organic cation transporters. CONCLUSIONS: These UDCA-HDAC6i conjugates open a therapeutic avenue for PLDs.

Current and novel therapeutic opportunities for systemic therapy in biliary cancer.

Biliary tract cancers (BTCs) are a group of rare and aggressive malignancies that arise in the biliary tree within and outside the liver. Beyond surgical resection, which is beneficial for only a small proportion of patients, current strategies for treating patients with BTCs include chemotherapy, as a single agent or combination regimens, in the adjuvant and palliative setting. Increased characterisation of the molecular landscape of these tumours has facilitated the identification of molecular vulnerabilities, such as IDH mutations and FGFR fusions, that can be exploited for the treatment of BTC patients. Beyond targeted therapies, active research avenues explore the development of novel therapeutics that target the crosstalk between cancer and stroma, the cellular pathways involved in the regulation of cell death, the chemoresistance phenotype and the dysregulation of RNA. In this review, we discuss the therapeutic opportunities currently available in the management of BTC patients, and explore the strategies that can support the implementation of precision oncology in BTCs, including novel molecular targets, liquid biopsies and patient-derived predictive tools.

Proteostasis disturbances and endoplasmic reticulum stress contribute to polycystic liver disease: New therapeutic targets.

BACKGROUND & AIMS: Polycystic liver diseases (PLDs) are genetic disorders characterized by progressive development of multiple biliary cysts. Recently, novel PLD-causative genes, encoding for endoplasmic reticulum (ER)-resident proteins involved in protein biogenesis and transport, were identified. We hypothesized that aberrant proteostasis contributes to PLD pathogenesis, representing a potential therapeutic target. METHODS: ER stress was analysed at transcriptional (qPCR), proteomic (mass spectrometry), morphological (transmission electron microscopy, TEM) and functional (proteasome activity) levels in different PLD models. The effect of ER stress inhibitors [4-phenylbutyric acid (4-PBA)] and/or activators [tunicamycin (TM)] was tested in polycystic (PCK) rats and cystic cholangiocytes in vitro. RESULTS: The expression levels of unfolded protein response (UPR) components were upregulated in liver tissue from PLD patients and PCK rats, as well as in primary cultures of human and rat cystic cholangiocytes, compared to normal controls. Cystic cholangiocytes showed altered proteomic profiles, mainly related to proteostasis (ie synthesis, folding, trafficking and degradation of proteins), marked enlargement of the ER lumen (by TEM) and hyperactivation of the proteasome. Notably, chronic treatment of PCK rats with 4-PBA decreased liver weight, as well as both liver and cystic volumes, of animals under baseline conditions or after TM administration compared to controls. In vitro, 4-PBA downregulated the expression (mRNA) of UPR effectors, normalized proteomic profiles related to protein synthesis, folding, trafficking and degradation and reduced the proteasome hyperactivity in cystic cholangiocytes, reducing their hyperproliferation and apoptosis. CONCLUSIONS: Restoration of proteostasis in cystic cholangiocytes with 4-PBA halts hepatic cystogenesis, emerging as a novel therapeutic strategy.

Survey of clenbuterol in bovine muscle and liver in Ecuador.

Clenbuterol is a steroid-type drug used in respiratory treatments in both humans and animals. However, it has a secondary effect related to the hypertrophy process in muscle and fat reduction. The illegal or bad use of clenbuterol has been reported in several countries, but there is scarce information in South America, where the production and consumption of meat are considerable. In this sense, the present study aimed at evaluating the occurrence of clenbuterol in bovine muscle and liver samples from a high cattle production area of Ecuador in 2015 and 2018. For this purpose, 57-58 samples were evaluated in 2015 and 20 samples in 2018 using the Enzyme-Linked Inmuno Sorbent Assay and ultrahigh-performance liquid chromatography-tandem mass spectrometry. The results showed complained results for clenbuterol in meat samples from both years and 23% (2015) and 85% (2018) of the samples of meat complied the maximum residue level defined by CODEX.

Correction to: Molecular characterization and complete genome of alstroemeria mosaic virus (AlMV).

The original version of this article unfortunately contained an error in the length of AIMV genome sequence.