Kappa-opioid receptor antagonism in the nucleus accumbens shell distinguishes escalated alcohol consumption and negative affective-like behavior from physiological withdrawal in alcohol-dependence.

Alcohol use disorder (AUD) is a chronic relapsing disease that is deleterious at individual, familial, and societal levels. Although AUD is one of the highest preventable causes of death in the USA, therapies for the treatment of AUD are not sufficient given the heterogeneity of the disorder and the limited number of approved medications. To provide better pharmacological strategies, it is important to understand the neurological underpinnings of AUD. Evidence implicates the endogenous dynorphin (DYN)/κ-opioid receptor (KOR) system recruitment in dysphoric and negative emotional states in AUD to promote maladaptive behavioral regulation. The nucleus accumbens shell (AcbSh), mediating motivational and emotional processes that is a component of the mesolimbic dopamine system and the extended amygdala, is an important site related to alcohol's reinforcing actions (both positive and negative) and neuroadaptations in the AcbSh DYN/KOR system have been documented to induce maladaptive symptoms in AUD. We have previously shown that in other nodes of the extended amygdala, site-specific KOR antagonism can distinguish different symptoms of alcohol dependence and withdrawal. In the current study, we examined the role of the KOR signaling in the AcbSh of male Wistar rats in operant alcohol self-administration, measures of negative affective-like behavior, and physiological symptoms during acute alcohol withdrawal in alcohol-dependence. To induce alcohol dependence, rats were exposed to chronic intermittent ethanol vapor for 14 h/day for three months, during which stable escalation of alcohol self-administration was achieved and pharmacological AcbSh KOR antagonism ensued. The results showed that AcbSh KOR antagonism significantly reduced escalated alcohol intake and negative affective-like states but did not alter somatic symptoms of withdrawal. Understanding the relative contribution of these different drivers is important to understand and inform therapeutic efficacy approaches in alcohol dependence and further emphasis the importance of the KOR/DYN system as a target for AUD therapeutics.

An afferent nerve-like electronic device with somatic mechanical perception and sensation management.

Tactile and pain perception are essential for biological skin to interact with the external environment. This complex interplay of sensations allows for the detection of potential threats and appropriate responses to stimuli. However, the challenge is to enable flexible electronics to respond to mechanical stimuli such as biological skin, and researchers have not clearly reported the successful integration of somatic mechanical perception and sensation management functions into neuro-like electronics. In this work, an afferent nerve-like device with a pressure sensor and a perception management module is proposed. The pressure sensor comprises two conductive fabric layers and an ionic hydrogel, forming a capacitor structure that emulates the swift transition from tactile to pain perception under mechanical stimulation. Drawing inspiration from the neuronal "gate control" mechanism, the sensation management module adjusts signals in response to rubbing, accelerating the discharge process and reducing the perception duration, thereby replicating the inhibitory effect of biological neurons on pain following tactile interference. This integrated device, encompassing somatic mechanical perception and sensation management, holds promise for applications in soft robotics, prosthetics, and human-machine interaction.

Assessment of In Vivo Chemical Mutagenesis by Long-Read Sequencing.

Evaluating the mutagenic properties of chemicals is crucial for understanding their potential cancer risks. Recent Illumina-based error-corrected sequencing techniques have enabled the direct detection of mutations induced de novo by mutagens. However, as the Illumina platform lacks intrinsic error-correction capabilities, complex library preparations and bioinformatic processes are necessary to identify these rare mutations. In this study, we evaluated whether long-read PacBio-based HiFi sequencing (HiFi seq), which has integrated error-correction, can detect de novo mutations induced by mutagens in C57BL/6 mouse tissues. Using HiFi seq, dose-dependent increases in mutation frequencies were found in tissues from mice exposed to 7,12-dimethylbenz[a]anthracene, procarbazine, and N-propyl-N-nitrosourea. Furthermore, the mutational signatures derived from these exposures were consistent with those previously reported for these mutagens. This study demonstrates that HiFi seq can complement established mutation detection assays to facilitate the identification of hazardous compounds.

Of criminals and cancer: The importance of social bonds and innate morality on cellular societies.

The current dogma in cancer biology contends that cancer is an identity problem: mutations in a cell's DNA cause it to "go rogue" and proliferate out of control. However, this largely ignores the role of cell-cell interaction and fails to explain phenomena such as cancer reversion, the existence of cancers without mutations, and foreign-body carcinogenesis. In this proof-of-concept paper, we draw on criminology to propose that cancer may alternatively be conceptualized as a relational problem: Although a cell's genetics is essential, the influence of its interaction with other cells is equally important in determining its phenotype. We create a simple agent-based network model of interactions among normal and cancer cells to demonstrate this idea. We find that both high mutation rates and low levels of connectivity among cells can promote oncogenesis. Viewing cancer as a breakdown in communication networks among cells in a tissue complements the gene-centric paradigm nicely and provides a novel perspective for understanding and treating cancer.

A study of somatization symptoms and low-frequency amplitude fluctuations of emotional memory in adolescent depression.

Studies have revealed that somatization symptoms are associated with emotional memory in adolescents with depressive disorders. This study investigated somatization symptoms and emotional memory among adolescents with depressive disorders using low-frequency amplitude fluctuations (ALFF). Participants were categorized into the somatization symptoms (FSS) group, non-FSS group and healthy control group (HC). The correctness of negative picture re-recognition was higher in the FFS and HC group than in the non-FSS group. The right superior occipital gyrus and right inferior temporal gyrus were significantly larger in the FSS group than those in the non-FSS and HC groups. Additionally, the ALFF in the superior occipital and inferior temporal gyrus were positively correlated with CSI score. Furthermore, the ALFF values in the temporal region positively correlated with correct negative image re-recognition. The negative image re-recognition rate was positively correlated with the ALFF in the left and right middle occipital gyri. These findings indicated that somatization symptoms in adolescent depression are associated with the superior occipital gyrus and inferior temporal gyrus. Notably, somatization symptoms play a role in memory bias within depressive disorders, with middle occipital and inferior temporal gyri potentially serving as significant brain regions.

Research on Depression in Children with Severe Bronchial Asthma: The Impact of Alexithymia and Somatic Symptoms.

Objective: The prevalence of depression in children with severe bronchial asthma is a significant concern due to its potential effects on illness burden and quality of life. This cross-sectional study aims to explore the relationship between depression and severe bronchial asthma in children, focusing on the impact of alexithymia and somatic symptoms. Methods: The study includes a total of 186 children aged 6-14 years diagnosed with severe bronchial asthma between 2008 and 2022 in our institute. Alexithymia was assessed using the Toronto Alexithymia Scale-20 items (TAS-20). Somatization symptoms were measured using the children's somatization inventory (CSI). The Hamilton depression scale (HAMD) was used to evaluate depression. Spearman correlation analysis was used to describe the correlation between alexithymia, somatization symptoms, and depression. Results: Children with bronchial asthma are found to have a significantly higher prevalence of depression, estimated to be around 16.67%. Approximately 98.92% of children exhibit varying degrees of somatic symptoms. Approximately 3.23% of children have alexithymia. The Spearman correlation analysis revealed that somatic symptoms and alexithymia were positive correlated with the depression. The correlation coefficients were 0.986 and 0.981 (P < .01), respectively. moreover, according to the results of multiple linear regression analysis, somatization symptoms and alexithymia significantly affects depression in children with severe bronchitis asthma (P < .01). Conclusion: These findings suggest that children with severe bronchial asthma experience a higher prevalence of depression, impacting their overall quality of life. In addition, the presence of somatic symptoms is prevalent among these children, further contributing to the burden on their quality of life. Moreover, somatization symptoms and alexithymia have been identified as a significant factor positive affecting depression in this population. Addressing these factors in clinical interventions may be beneficial for improving the overall well-being in this population.

Application of cold plasma therapy for managing subclinical mastitis in cows induced by Streptococcus agalactiae, Streptococcus uberis and Escherichia coli.

The primary objective of this study was to assess the effectiveness of cold plasma therapy in managing subclinical mastitis in cows caused by Streptococcus agalactiae, Streptococcus uberis and Escherichia coli. After detection of mastitis pathogens, 38 cows were selected for cold plasma therapy for five days. On the fifth day of treatment, the mastitis agents were re-examined and no causative agents were identified. An additional evaluation conducted 28 days later confirmed the absence of mastitis. Cow productivity, milk composition and quality indicators were assessed at the beginning of the experiment and 32 days from the start (28 days after treatment cessation). After the mastitis treatment, the somatic cell count decreased significantly by between 2.89 and 7.09 times, and the milk yield of the cows at the end of the experiment increased from 0.63 kg per day to 2.82 kg per day (P < 0.01). These results highlight the potential of this innovative approach for managing a prevalent disease that causes substantial losses in the dairy industry. Furthermore, they lay the groundwork for expanded research involving larger sample sizes.

Embryonic temperature has long-term effects on muscle circRNA expression and somatic growth in Nile tilapia.

Embryonic temperature has a lasting impact on muscle phenotype in vertebrates, involving complex molecular mechanisms that encompass both protein-coding and non-coding genes. Circular RNAs (circRNAs) are a class of regulatory RNAs that play important roles in various biological processes, but the effect of variable thermal conditions on the circRNA transcriptome and its long-term impact on muscle growth plasticity remains largely unexplored. To fill this knowledge gap, we performed a transcriptomic analysis of circRNAs in fast muscle of Nile tilapia (Oreochromis niloticus) subjected to different embryonic temperatures (24°C, 28°C and 32°C) and then reared at a common temperature (28°C) for 4 months. Nile tilapia embryos exhibited faster development and subsequently higher long-term growth at 32°C compared to those reared at 28°C and 24°C. Next-generation sequencing data revealed a total of 5,141 unique circRNAs across all temperature groups, of which 1,604, 1,531, and 1,169 circRNAs were exclusively found in the 24°C, 28°C and 32°C groups, respectively. Among them, circNexn exhibited a 1.7-fold (log2) upregulation in the 24°C group and a 1.3-fold (log2) upregulation in the 32°C group when compared to the 28°C group. Conversely, circTTN and circTTN_b were downregulated in the 24°C groups compared to their 28°C and 32°C counterparts. Furthermore, these differentially expressed circRNAs were found to have multiple interactions with myomiRs, highlighting their potential as promising candidates for further investigation in the context of muscle growth plasticity. Taken together, our findings provide new insights into the molecular mechanisms that may underlie muscle growth plasticity in response to thermal variation in fish, with important implications in the context of climate change, fisheries and aquaculture.

Pelvic Palpatory Tests in Manual Therapy and Osteopathy: A Critical Review of the Literature and Suggestions for New Research.

Manual therapists apply physical interventions to the entire structure of the body to promote healing, prevent pathologies, and improve patient health. In osteopathic practice, palpatory evaluation is considered a fundamental clinical practice requirement for identifying somatic dysfunction. Most of the articles published in this area have failed to demonstrate a level of reproducibility that supports palpation in evidence-based clinical practice. When considering the poor reliability of the palpatory tests highlighted in the literature, a discrepancy is noted with what is known about the tactile sensitivity of human hands. For static touch, the minimum size that can be detected, in the absence of applied movement or vibration, is approximately 0.2 mm. Yet, it seems that this high level of precision is insufficient to ensure reliability in the tests used to evaluate osteopathic somatic dysfunction. The purpose that underscores this article is to determine how these two contradictory elements, high sensitivity and low reliability, can be present in palpatory tests. The article reports the literature findings regarding palpatory tests of pelvic, which is an important structure for clinical purposes. Additionally, a critical review of how these studies were conducted is provided to identify any elements that may justify the obtained results. Following recent accredited guidelines present in the literature, we propose suggestions on vision training methods, manual perception refinement training, the search for anatomical markers, and the position of the examiner in relation to the examinee that may be useful for future studies on the topic covered by the article.

Neuropsychological features of somatic symptom disorder and depression/anxiety in Taiwan: An analysis based on the comorbidity status.

Individuals with somatic symptom disorder (SSD) often have comorbid depression or anxiety, but whether SSD is associated with specific neuropsychological functions has yet to be fully examined. We analyzed which neuropsychological features are more closely associated with SSD, anxiety, and depression. In this case-control study, we recruited 140 individuals with SSD, 104 individuals with affective disorders without SSD, and 159 healthy controls in Taiwan. We collected DSM-5 diagnoses, questionnaire scores, and performance on eight tasks from the Cambridge Neuropsychological Test Automated Battery (CANTAB) for each participant. Several CANTAB tasks involving attention, executive function, and social cognition showed significant group differences. In the adjusted analysis, the tasks significantly associated with SSD were the Match to Sample Visual Search (MTS) and the Emotion Recognition Task (ERT). Among the questionnaires, the Cognitions about Body and Health Questionnaire showed the most significant associations with the tasks, specifically with Rapid Visual Information Processing, MTS, Paired Associates Learning, Spatial Working Memory, Intra-Extra Dimensional Set Shift, and ERT. We conclude that the MTS and ERT tasks show significant relationships with both SSD diagnosis and related questionnaires. These tasks primarily involve selective attention and negative emotion regulation.

The Influence of Selected Environmental Factors on the Number of Somatic Cells in Cistern and Alveolar Milk of Polish Holstein-Friesian Cows.

The aim of the study was to evaluate the influence of the milking phase on somatic cell count (SCC) in milk obtained from the cisternal and alveolar parts of udders of selected Polish Holstein-Friesian cows. The study also assessed the impact of other genetic and environmental factors on SCC variability in cisternal and alveolar milk, including: the individual cow, lactation stage, age of cow, production level, milking speed, fat-to-protein ratio, and milking type. The research included 15 cows of Polish Holstein-Friesian breed at different ages, lactation stages, and with varying daily milk yield. A total of 210 milk observations were conducted, including 105 for 1 min milking and 105 for 8 min milking. The results obtained in the study indicated that milk obtained during two different milking phases exhibited similar SCC levels (F for LOGSCC = 0.79). The average actual SCC in milk produced by 15 cows in 105 observations for 1 min milking was 219,000 cells/mL, while for 8 min milking it was 229,000 cells/mL. The results were inconclusive, suggesting that SCC in cisternal and alveolar milk must be influenced by factors other than the milking phase. The analysis of variance conducted for this purpose provided the basis for stating a highly statistically significant effect of the individual cow (F for LOGSCC = 147.9), lactation stage (F for LOGSCC = 54.64), age of cow (F for LOGSCC = 12.39), daily production level (F for LOGSCC = 34.49), milking speed (F for LOGSCC = 17.56), and fat-to-protein ratio (F for LOGSCC = 22.99) on the variability of characteristics defining SCC in milk. In summary, SCC is characterized by high variability, influenced by a range of environmental and genetic factors such as the individual cow, lactation stage, age of cow, milking speed, and dietary fat-to-protein ratio. The influence of milking phase (1 min or 8 min) and milking type (morning or evening) should be considered inconclusive based on the entire population studied. For half of the cows, SCC in cisternal milk was higher than in alveolar milk, while for the other half, the situation was reversed. Further observations are required to confirm the hypothesis regarding the extent to which cows' immunological response to bacterial infections is concentrated in the cisternal or alveolar part of the udder under national environmental conditions.

Genetic diversification of allohexaploid Brassica hybrids (AABBCC) using a fertile octoploid with excessive C genome set (AABBCCCC).

MAIN CONCLUSION: Using octoploid somatic hybrids with excessive C genome sets, AABBCCCC, a diverse allohexaploid, AABBCC, was produced by C genome reduction through subsequent crossing with various AABB cultivars. Even when somatic hybrids are produced, the plants that are produced are rarely in themselves an innovative crop. In this study, we used somatic hybrids of Brassica juncea (AABB) and B. oleracea (CC) as model cases for the genetic diversification of the somatic hybrids. One cell of 'Akaoba Takana' (B. juncea) and two cells of 'Snow Crown' (B. oleracea) were fused to create several somatic hybrids with excessive C genomes, AABBCCCC. Using AABBCCCC somatic hybrids as mother plants and crossing with 'Akaoba Takana', the AABBCC progenies were generated. When these AABBCC plants were self-fertilized, and flow cytometric (FCM) analysis was performed on the next generations, differences in the relative amount of genome size variation were observed, depending on the different AABBCCCC parents used for AABBCC creation. Further self-progeny was obtained for AABBCC plants with a theoretical allohexaploid DNA index by FCM. However, as the DNA indices of the progeny populations varied between plants used and aneuploid individuals still occurred in the progeny populations, it was difficult to say that the allohexaploid genome was fully stabilized. Next, to obtain genetic diversification of the allohexaploid, different cultivars of B. juncea were crossed with AABBCCCC, resulting in diverse AABBCC plants. Genetic diversity can be further expanded by crossbreeding plants with different AABBCC genome sets. Although genetic stability is necessary to ensure in the later generations, the results obtained in this study show that the use of somatic hybrids with excess genomes is an effective strategy for creating innovative crops.

Subjective cognitive concerns not related to objective impairment in patients with somatic symptom and related disorders.

OBJECTIVES: Patients with Somatic Symptom and Related Disorders (SSRD) report subjective cognitive concerns, and research indicates that they show objective cognitive impairment. This study explored the value of subjective concerns flagging objective impairment. Furthermore, we investigated whether coping moderated this relationship, and the role of depressive symptomatology. METHOD: In a cross-sectional design, objective impairment was measured with an extensive neuropsychological assessment; subjective concerns with the Cognitive Failure Questionnaire; coping styles with the Coping Inventory of Stressful Situations; and symptoms of depression with the Patient Health Questionnaire- 9. RESULTS: The results show that subjective concerns are of limited value in signaling objective impairment in patients with SSRD. Regression analyses performed on data from 225 patients showed that symptoms of depression (ß = .32) were the main predictor of subjective concerns, which were unrelated to objective impairment. Coping was not a moderator, but patients with emotion-oriented coping styles had more subjective concerns (ß=.40), and conversely, patients with avoidance- and/or task-oriented coping styles had less (respectively, ß=-.27 and ß=-.24). CONCLUSIONS: These results align with the Somatosensory Amplification Theory; patients with SSRD may amplify benign cognitive failures and experience them as intrusive, noxious, and more intense. In patients with SSRD, subjective cognitive concerns are more related to psychological constructs (symptoms of depression and coping styles) than to objective impairment.

Delusion of pregnancy: Case series.

Delusion of pregnancy is a relatively uncommon manifestation in psychotic patients. Differentiating it from closely related clinical entities like pseudocyesis/pseudopregnancy can be a challenge in clinical setting. Diagnosis of a psychotic illness with delusional pregnancy as the presenting feature in the absence of other florid symptoms of psychosis is a different challenge altogether. Currently, there is no consensus on the pharmacotherapy and psychotherapy of delusional pregnancy. We present a case series on delusional pregnancy with the intent to answer some of these gaps in the literature.

An investigation of antioxidative and anti-inflammatory effects of Taraxacum coreanum (white dandelion) in lactating Holstein dairy cows.

Objective: The aim of this investigation was to examine the impact of Taraxacum coreanum (known as dandelion) (TC) and TC mixtures with milk thistle (MT) or Aspergillus oryzae (AO) as feed additives on the immune response, milk quality, and milk production in Holstein cows over 6 weeks of administration. Materials and Methods: Thirty-two healthy Holstein dairy cows were provided 30 kg of total mixed ration (TMR) with no TC, 90 gm TC, 54 gm TC + 36 gm MT, or 54 gm TC + 36 gm AO 40% groups. The feed additives were supplied daily in two equal portions (per 45 gm) by topdressing the TMR for 6 weeks. Milk and blood samples were collected weekly. Results: In the TC-treated cows (TC, TC + MT, and TC + AO groups), significantly lower peripheral blood white blood cell (WBC) counts at 6 weeks and milk somatic cell counts (SCCs) at 4-6 weeks of administration were observed. Concentrations of serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) were notably elevated in cows treated with TC for 4-6 weeks, while levels of proinflammatory cytokines concentrations of tumor necrosis factor-alpha (TNF-α) and chemokine (IL-8) were significantly reduced in TC-treated cows after 3-6 weeks of administration. Conclusion: These results suggested that TC or a TC mixture with other medicinal herbs supplementations enhanced the serum antioxidative activities and, consequently, might suppress the adverse immune response due to lower serum TNF-α and IL-8 release supported by lower WBC and SCC counts.

A systematic cognitive behavioral therapy approach for pediatric disorders of gut-brain interaction.

OBJECTIVE: Cognitive Behavioral Therapy (CBT) for youth with Disorders of Gut-Brain Interaction (DGBIs) is effective; however, there are calls in the field to strengthen the evidence base and identify specific mechanisms of treatment that yield the most benefit for this patient population. A unique, systematic treatment approach of CBT with initial evidence for success for pediatric patients with DGBIs was evaluated to further demonstrate its clinical utility in this population. METHODS: This was a retrospective study of 42 pediatric patients aged 11-17 years with DGBIs, who were diagnosed and referred for CBT by pediatric gastroenterology providers. Providers also completed a survey rating acceptability and effectiveness of CBT. The systematic CBT approach included 10 sessions delivered by a psychologist at an integrated Pediatric GI Clinic. RESULTS: Review of 42 pediatric charts showed significant decreases in self-reported functional disability, abdominal pain, as well as depression and anxiety symptoms pre- to post-CBT completion. A moderation effect was observed where patients reporting higher levels of depressive symptoms and primary symptom of abdominal pain reported smaller reductions in functional impairment compared to those with lower levels of depression and primary symptom of nausea or vomiting. Pediatric Gastroenterology providers were satisfied with this psychological treatment approach. CONCLUSIONS: This study provides evidence for acceptability and effectiveness of implementation of a systematic CBT approach for pediatric DGBIs in an integrated GI clinic, as well as areas worthy of future research, including identifying the most important mechanisms of treatment and factors that influence treatment response.

Propagation path of a flowering cherry (Cerasus × yedoensis) cultivar 'Somei-Yoshino' traced by somatic mutations.

In the long history of human relations with flowering cherry trees in Japan, 'Somei-Yoshino' occupies an exceptional position among a variety of flowering trees: it is a self-incompatible interspecific hybrid but has been enthusiastically planted by grafting throughout Japan, due most likely to its flamboyant appearance upon full bloom. Thus, 'Somei-Yoshino' gives us a rare opportunity to trace and investigate the occurrence and distribution of somatic mutations within a single plant species through analysis of the genomes of the clonally propagated trees grown under a variety of geographical and artificial environments. In the studies presented here, a total of 46 samples of 'Somei-Yoshino' trees were collected and their genomes were analyzed. We identified 684 single nucleotide mutations, of which 71 were present in more than two samples. Clustering analysis of the mutations indicated that the 46 samples were classified into eight groups, four of which included 36 of the 46 samples analyzed. Interestingly, all the four tree samples collected in Ueno Park of Tokyo were members of the four groups mentioned above. Based on comparative analysis of their mutations, one of the four trees growing in Ueno Park was concluded to be the closest to the original ancestor. We propose that somatic mutations may be used as tracers to establish the ancestral relationship amongst clonally propagated individuals.

Generation of Human-Induced Pluripotent Stem Cells from Peripheral Blood Mononuclear Cells of C9ORF72-Associated Amyotrophic Lateral Sclerosis Patients.

Disease modeling of neuromuscular disorders, such as amyotrophic lateral sclerosis (ALS), is hindered by limited accessibility of affected cells. This problem can be overcome by generation of human induced pluripotent stem cells (hiPSC), which can be then differentiated into required cells. Here, we describe the detailed protocol of hiPSC establishment from peripheral blood mononuclear cells (PBMC) of two ALS patients with detected expansion of G4C2 (GGGGCC) repeats in the first intron of C9ORF72 gene, known to be linked with the most common form of familial ALS.Successful PBMC reprogramming with non-integrating Sendai vectors was confirmed by expression of pluripotency markers: OCT4, NANOG, SSEA4, and TRA-1-60 in obtained hiPSC and their ability to differentiate into cells of three germ layers.The generated ALS-patient-specific hiPSC create a possibility for deciphering molecular basis of this devastating neuromuscular disease.

Duodenal Fluid Analysis as a Rewarding Approach to Detect Low-Abundance Mutations in Biliopancreatic Cancers.

Diagnosis of biliopancreatic cancers by the available serum tumor markers, imaging, and histopathological tissue specimen examination remains a challenge. Circulating cell-free DNA derived from matched pairs of secretin-stimulated duodenal fluid (DF) and plasma from 10 patients with biliopancreatic diseases and 8 control subjects was analyzed using AmpliSeq™ HD technology for Ion Torrent Next-Generation Sequencing to evaluate the potential of liquid biopsy with DF in biliopancreatic cancers. The median cfDNA concentration was greater in DF-derived than in plasma-derived samples. A total of 13 variants were detected: 11 vs. 1 were exclusive for DF relative to the plasma source, and 1 was shared between the two body fluids. According to the four-tier systems, 10 clinical tier-I-II (76.9%), 1 tier-III (7.7%), and 2 tier-IV (15.4%) variants were identified. Notably, the 11 tier-I-III variants were exclusively found in DF-derived cfDNA from five patients with biliopancreatic cancers, and were detected in seven genes (KRAS, TP53, BRAF, CDKN2A, RNF43, GNAS, and PIK3CA); 82% of the tier-I-III variants had a low abundance, with a VAF < 6%. The mutational profiling of DF seems to be a reliable and promising tool for identifying cancer-associated alterations in malignant cancers of the biliopancreatic tract.