Molecular tumor board: molecularly adjusted therapy upon identification and functional validation of a novel ALK resistance mutation in a case of lung adenocarcinoma.

We report a case of a long-term surviving patient with EML4/ALK translocated non-small cell adenocarcinoma of the lung in UICC8 stage IVA. During recurrence under continuous crizotinib therapy, a hitherto insufficiently characterized missense mutation in the ALK gene (Arg1181His) was identified through targeted sequencing. The aforementioned EML4/ALK translocation could still be detected in this situation. Employing a 3D reconstruction of the ALK tertiary structure, considering its interaction with various ALK inhibitors at the molecular binding site, our analysis indicated the presence of a mutation associated with crizotinib resistance. To validate the biological relevance of this previously unknown mutation, we carried out an in vitro validation approach in cell culture in addition to the molecular diagnostics accompanied by the molecular tumor board. The tumor scenario was mimicked through retroviral transfection. Our comparative in vitro treatment regimen paired with the clinical trajectory of the patient, corroborated our initial clinical and biochemical suspicions. Our approach demonstrates preclinical, in silico, and clinical evidence of a novel crizotinib resistance mutation in ALK as well as sensitivity toward brigatinib and potentially lorlatinib. In future cases, this procedure represents an important contribution to functional diagnostics in the context of molecular tumor boards.

Late Onset of Primary Hemophagocytic Lymphohistiocytosis (HLH) with a Novel Constellation of Compound Heterozygosity Involving Two Missense Variants in the PRF1 Gene.

Hemophagocytic lymphohistiocytosis (HLH) is a rare but in most cases life-threatening immune-mediated disease of the hematopoietic system frequently associated with hematologic neoplasms. Here, we report on a case in which we detected a novel constellation of two missense variants affecting the PRF1 gene, leading to de novo primary HLH. Diagnostics included a comprehensive clinical work-up and standard methods of hematopathology as well as extended molecular genomics based on polymerase chain reaction (PCR) reactions and the calculation of three-dimensional molecule reconstructions of PRF1. Subsequently, a comprehensive review of the literature was performed, which showed that this compound heterozygosity has not been previously described. The patient was a 20-year-old female. Molecular diagnostics revealed two heterozygous missense variants in the PRF1 gene (A91V and R104C) on exon 2. Apart from the finding of two inconclusive genetic variants, all clinical criteria defined by the HLH study group of Histiocyte Society were met at initial presentation. The final diagnosis was made in cooperation with the Consortium of German HLH-reference centers. Here, chemotherapy did not lead to sufficient sustained disease control. Therefore, the decision for allogenic hematopoietic stem cell transplantation (alloHSCT) was made. Hitherto, the duration of response was 6 months. Due to severe and unmanageable hepatic graft-versus-host disease (GvHD), the patient died. We report on a novel constellation of a compound heterozygosity containing two missense variants on exon 2 of the PRF1 gene. To the authors' best knowledge, this is the first presentation of a primary HLH case harboring this genomic constellation with late-onset clinical manifestation.

Temporal changes in metal and arsenic concentrations in blood and feathers of tropical seabirds after one of the largest environmental disasters associated with mining.

Monitoring of contaminant levels in wildlife over time is a tool for assessing the presence and persistence of environmental impacts at ecosystem, community and population levels. Tropical seabirds breeding in the Abrolhos Archipelago, 70 km off the Brazilian coast, forage in areas under the influence of the Doce River discharge. In 2015, the Fundão Dam collapsed and released ca 60 million tons of iron ore tailings into the ocean. In the present study, red-billed tropicbirds Phaethon aethereus and brown boobies Sula leucogaster breeding in Abrolhos were monitored over four years (2019-2022) for metal (Fe, Mn, Zn, Cu, Cr, Hg, Pb, Cd) and metalloid (As) concentrations in blood and feathers. Over six sampling events, metal (loid) concentrations showed strong temporal variation in both tissues. Overall, feathers showed greater element concentrations than blood, with stronger correlations between elements, especially Mn and the nonessential As, Cd, Hg and Pb. Mn is one of the major chemical markers of the Fundão Dam tailings. Metal (loid) concentrations in the tropical seabirds evaluated were above suggested threshold levels for most nonessential elements (As, Cd and Pb), especially in February 2021, when metal (loid) concentrations peaked in feathers. In this case, values were orders of magnitude higher than those observed in other sampling events. This occurred one year after a major rainy season in the Doce River basin, which increased river discharge of contaminated mud into the ocean, where contaminants are further remobilized by winds and currents, resulting in transference through the marine food web. This finding is consistent to what has been observed for other ecosystem compartments monitored in the region under the influence of the Doce River. Our findings highlight the utility of using tropical seabirds as sentinels of marine pollution, revealing strong temporal patterns in metal (loid) concentrations associated to bottom-up climatic processes.

Assessment of Human Health Risk of Metal(Loid) Content in Brazilian Sardine Along the Southwestern Atlantic.

The Brazilian sardine (Sardinella aurita) is an important food resource found in the subtropical Southwestern Atlantic Continental Shelf (CSSWA), but limited information about its metal(loid) concentrations is available, restricting effective risk assessment by its consumption. On this research, we hypothesized that S. aurita presents different metal(loid) concentrations within a latitudinal gradient in the CSSWA (northern and southern sectors). We also assessed the S. aurita consumption contamination risk in both sectors of the CSSWA. The results indicated that S. aurita might present different chemical and contamination profiles between the observed sectors, with highlights to As, Cr, and Fe at greater levels than the safety limits established by regulatory agencies. Such finds could be explained by urbanization, industrialization, continental and oceanographic processes along the CSSWA, corroborating our hypothesis for most observed metals(loid). On the other hand, our risk assessment analysis of metal(loid) concentrations did not indicate hazards for human consumption.

The expression of the adenosine pathway markers CD39 and CD73 in salivary gland carcinomas harbors the potential for novel immune checkpoint inhibition.

BACKGROUND: In salivary gland carcinomas (SGC), there is only a small fraction of entities that appears to profit from immune checkpoint inhibition (ICI). Recent findings connected the activation of adenosine-signaling with a tolerogenic microenvironment. Therefore, the inhibition of adenosine pathway markers (CD39 and/or CD73) can augment ICI and/or display a novel immunotherapeutic strategy beyond ICI. Here, we assessed the immuno-histochemical expression of CD39 and CD73 across a wide spectrum of SGCs. METHODS: In total, 114 patients with SGCs consecutively diagnosed between 2001 and 2021 were assessed for clinicopathological baseline characteristics and underwent confirmatory histopathological review. Immunohistochemical expression levels of CD39 and CD73 were assessed by applying the tumor proportion score (TPS) and the immune proportional score (IPS) comparable to PD-L1 expression analysis in routine clinical practice. Additionally, findings were correlated with PD-L1 expression levels. RESULTS: The median age was 60.6 and 51.8% patients were female. The cohort covered a spectrum of eight distinct entities. Advanced-stage disease (UICC/AJCC III/IVA-IVC) at initial diagnosis was present in the majority of patients (64/114). Immunohistochemical staining revealed positivity for CD39 and CD73 in 48.2% and 21.1% on tumor cells (TPS ≥ 1%) as well as 46.4% and 42.9% within the immune cell infiltrate (IPS ≥ 1%), respectively. Further comparative analyses revealed immune-cold entities such adenoid cystic carcinoma (AdCC), immune-hot tumors such as adenocarcinoma, not otherwise specified (AC (NOS)) and entities with intermediate immunologic features such as acinic cell carcinoma (ACC). CONCLUSION: Current results indicate entity-specific adenosine signaling signatures. These findings suggest that the adenosine pathway plays a decisive role in tumor immunity among the major spectrum of SGCs. Targeting the adenosine pathway might pose a promising therapeutic option for selected entities.

The Glasgow Prognostic Score Predicts Survival Outcomes in Neuroendocrine Neoplasms of the Gastro-Entero-Pancreatic (GEP-NEN) System.

Background: Across a variety of solid tumors, prognostic implications of nutritional and inflammation-based risk scores have been identified as a complementary resource of risk stratification. Methods: In this retrospective study, we performed a comparative analysis of several established risk scores and ratios, such as the Glasgow Prognostic Score (GPS), in neuroendocrine neoplasms of the gastro−entero−pancreatic (GEP-NEN) system with respect to their prognostic capabilities. Clinicopathological and treatment-related data for 102 GEP-NEN patients administered to the participating institutions between 2011 and 2021 were collected. Scores/ratios significantly associated with overall or progression-free survival (OS, PFS) upon univariate analysis were subsequently included in a Cox-proportional hazard model for the multivariate analysis. Results: The median age was 62 years (range 18−95 years) and the median follow-up period spanned 51 months. Pancreatic or intestinal localization at the initial diagnosis were present in 41 (40.2%) and 44 (43.1%) cases, respectively. In 17 patients (16.7%), the primary manifestation could not be ascertained (NNUP; neuroendocrine neoplasms of unknown primary). Histological grading (HG) revealed 24/102 (23.5%) NET/NEC (poorly differentiated; high grade G3) and 78/102 (76.5%) NET (highly or moderately differentiated; low−high grade G1−G2). In total, 53/102 (51.9%) patients presented with metastatic disease (UICC IV), 11/102 (10.7%) patients presented with multifocal disease, and 56/102 (54.9%) patients underwent a primary surgical or endoscopic approach, whereas 28 (27.5%) patients received systemic cytoreductive treatment. The univariate analysis revealed the GPS and PI (prognostic index), as well as UICC-stage IV, HG, and the Charlson comorbidity index (CCI) to predict both the PFS and OS in GEP-NEN patients. However, the calculation of the survival did not separate GPS subgroups at lower risk (GPS 0 versus GPS 1). Upon the subsequent multivariate analysis, GPS was the only independent predictor of both OS (p < 0.0001; HR = 3.459, 95% CI = 1.263−6.322) and PFS (p < 0.003; HR = 2.119, 95% CI = 0.944−4.265). Conclusion: In line with previous results for other entities, the present study revealed the GPS at baseline to be the only independent predictor of survival across all stages of GEP-NEN, and thus supports its clinical utility for risk stratification in this group of patients.

Pendulum-Based River Current Energy Converter for Hydrometric Monitoring Systems.

Energy harvesting from flowing water is important for supplying hydrometric monitoring systems. Nevertheless, it is challenging due to the chaotic water flow in only one main direction and the relatively weak energy profile. In this paper, a novel energy harvester has been proposed, designed, and validated. The converter consists of a pendulum, a gearbox, two overrunning clutches, a spiral spring, and a generator. By coupling the kinetic energy via an oscillating mass equipped with a magnetic spring, it is possible to accommodate the power supply, electronics, and sensors with data transmission in a completely closed, encapsulated, stable housing without an interface to the outside. In addition, an energy management circuit and a battery charging circuit were developed that could be housed in the sealed enclosure. The pendulum transducer prototype was tested with the developed online hydrometric measurement station, which consists of a multi-channel data logger with a cellular modem and a tipping bucket rain gauge sensor. The overall system was successfully validated by experimental studies in a river.

Comparative analysis of international prognostic indices in gray-zone lymphoma.

Gray-zone lymphoma (GZL) reflects an aggressive B-cell neoplasm with features intermediate between diffuse large B-cell lymphoma (DLBCL) and classical Hodgkin lymphoma (cHL). The International Prognostic Index (IPI) and its derivatives (R-IPI, NCCN-IPI, and the Hasenclever IPS) have been established for DLBCL or cHL while the most suitable scoring system for GZL remains undetermined. In an exploratory multi-centric cohort of GZL (n = 61), we performed a comparative analysis of prognostic indices with regard to model fit and mutual concordance. The calculation of the corrected Akaike's information criterion (cAIC) and Harrel's concordance index (c-index) for each scoring system identified the NCCN-IPI to harbor the most convincing prognostic capabilities regarding both overall survival (OS) and progression-free survival (PFS) compared to its enhanced derivatives. The current results affirm the clinical utility of the NCCN-IPI and suggest its preferential use in clinical practice in GZL-patients.

Prognostic impact of nutritional and inflammation-based risk scores in follicular lymphoma in the era of anti-CD20 targeted treatment strategies.

BACKGROUND: The composition of the tumor microenvironment (TME) is conditioned by immunity and the inflammatory response. Nutritional and inflammation-based risk scores have emerged as relevant predictors of survival outcome across a variety of hematological malignancies. METHODS: In this retrospective multicenter trial, we ascertained the prognostic impact of established nutritional and inflammation-based risk scores [Glasgow Prognostic Score (GPS), C-reactive-protein/albumin ratio (CAR), neutrophil-lymphocyte ratio (NLR), prognostic nutritional index (PNI), and prognostic index (PI)] in 209 eligible patients with histologically confirmed CD20+ follicular lymphoma (FL) of WHO grade 1 (37.3%), 1-2 (16.3%), 2 (26.8%) or 3A (19.8%) admitted to the participating centers between January 2000 and December 2019. Characteristics significantly associated with overall or progression-free survival (OS, PFS) upon univariate analysis were subsequently included in a Cox proportional hazard model. RESULTS: In the study cohort, the median age was 63 (range 22-90 years). The median follow-up period covered 99 months. The GPS and the CAR were identified to predict survival in FL patients. The GPS was the only independent predictor of OS (p < 0.0001; HR 2.773; 95% CI 1.630-4.719) and PFS (p = 0.001; HR 1.995; 95% CI 1.352-2.944) upon multivariate analysis. Additionally, there was frequent occurrence of progression of disease within 24 months (POD24) in FL patients with a calculated GPS of 2. CONCLUSION: The current results indicate that the GPS predicts especially OS in FL patients. Moreover, GPS was found to display disease-specific effects in regard to FL progression. These findings and potential combinations with additional established prognosticators should be further validated within prospective clinical trials.

A new species of viviparous brotula genus Pseudogilbia (Ophidiiformes: Dinematichthyidae) from Brazilian reefs, with an updated diagnosis of the genus.

In this study, a new species of Pseudogilbia Møller, Schwarzhans & Nielsen 2004 is described based on two male specimens (40-44 mm LS ) from shallow reefs of Bahia, Brazil. Pseudogilbia australis sp. nov. is distinguished from its only congener, Pseudogilbia sanblasensis Møller, Schwarzhans & Nielsen 2004 from Caribbean Panama, by having: two lower preopercular pores (vs. one); dorsal-fin rays 65-67 (vs. 69); anal-fin rays 51-53 (vs. 56); pectoral-fin rays 18 (vs. 20); caudal vertebrae 27-28 (vs. 30); pectoral-fin length 15.0%-15.9% LS (vs. 14.3); pelvic-fin length 13.5% LS (vs. 16.4) and a different morphology of the male copulatory organ. Pseudogilbia australis sp. nov. is the only dinematichthyid so far recorded in the South Atlantic. An updated diagnosis for the genus is also provided.

Mercury Concentrations in Four Marine Fishery Resources from Rio de Janeiro Coast, SW Atlantic, and Potential Human Health Risk Via Fish Consumption.

Mercury (Hg) contamination has caused ecological and socioeconomic problems worldwide. One of the main Hg contamination routes by the human populations is through fish ingestion. Herein, we analyzed the total Hg concentrations (THg) in highly consumed marine fish species: Brazilian flathead Percophis brasiliensis, Atlantic bigeye Priacanthus arenatus, Stripped weakfish Cynoscion guatucupa, and Jamaica weakfish C. jamaicensis. The specimens were collected from fishing landings off the Rio de Janeiro, SW Atlantic. Additionally, we calculate the estimated weekly intakes (EWI) and the maximum amount of fish (MAF) that can be safely ingested, both based on the provisional tolerable weekly intake (PTWI). The highest THg concentrations were recorded in P. brasiliensis with a mean of 340.59 µg/kg (dry weight) and the lowest in P. arenatus (115.75 µg/kg). THg concentrations were positively related to the δ15N ratios indicating an increase in contamination with trophic level. All samples showed THg concentrations below the established limits by Brazilian and international regulation. Hg intake via human fish consumption does not exceed the PTWI. The EWI varied from 1.41% (P. arenatus size class I) to 11.52% (P. brasiliensis class II) of the PTWI, while the estimated EWI for "fish consumers" varied between 8.35 and 68.07% PTWI. The MAF estimated for an adult (70 kg) could safely consume between 1277.85 and 7075.50 g/week. This study is an important baseline for monitoring and future comparisons. Therefore, it is important to maintain monitoring of mercury levels in fish species in different marine regions, especially those species most consumed by humans.

The southernmost records of Brotula barbata (Bloch & Schneider 1801) (Ophidiiformes: Ophidiidae) in the western Atlantic Ocean.

The present study reports the occurrence of the bearded brotula, Brotula barbata, in four different localities off southern Brazil. These records extend the species' known distribution limit southwards, beyond Rio de Janeiro, to the state of Rio Grande do Sul (30° 50' S). Morphometric and meristic data for the specimens reported herein are provided and discussed.

Merging national forest and national forest health inventories to obtain an integrated forest resource inventory--experiences from Bavaria, Slovenia and Sweden.

BACKGROUNDS, MATERIAL AND METHODS: To meet the demands of sustainable forest management and international commitments, European nations have designed a variety of forest-monitoring systems for specific needs. While the majority of countries are committed to independent, single-purpose inventorying, a minority of countries have merged their single-purpose forest inventory systems into integrated forest resource inventories. The statistical efficiencies of the Bavarian, Slovene and Swedish integrated forest resource inventory designs are investigated with the various statistical parameters of the variables of growing stock volume, shares of damaged trees, and deadwood volume. The parameters are derived by using the estimators for the given inventory designs. The required sample sizes are derived via the general formula for non-stratified independent samples and via statistical power analyses. The cost effectiveness of the designs is compared via two simple cost effectiveness ratios. RESULTS: In terms of precision, the most illustrative parameters of the variables are relative standard errors; their values range between 1% and 3% if the variables' variations are low (s%<80%) and are higher in the case of higher variations. A comparison of the actual and required sample sizes shows that the actual sample sizes were deliberately set high to provide precise estimates for the majority of variables and strata. In turn, the successive inventories are statistically efficient, because they allow detecting the mean changes of variables with powers higher than 90%; the highest precision is attained for the changes of growing stock volume and the lowest for the changes of the shares of damaged trees. Two indicators of cost effectiveness also show that the time input spent for measuring one variable decreases with the complexity of inventories. CONCLUSION: There is an increasing need for credible information on forest resources to be used for decision making and national and international policy making. Such information can be cost-efficiently provided through integrated forest resource inventories.