Seven quick tips for gene-focused computational pangenomic analysis.

Pangenomics is a relatively new scientific field which investigates the union of all the genomes of a clade. The word pan means everything in ancient Greek; the term pangenomics originally regarded genomes of bacteria and was later intended to refer to human genomes as well. Modern bioinformatics offers several tools to analyze pangenomics data, paving the way to an emerging field that we can call computational pangenomics. Current computational power available for the bioinformatics community has made computational pangenomic analyses easy to perform, but this higher accessibility to pangenomics analysis also increases the chances to make mistakes and to produce misleading or inflated results, especially by beginners. To handle this problem, we present here a few quick tips for efficient and correct computational pangenomic analyses with a focus on bacterial pangenomics, by describing common mistakes to avoid and experienced best practices to follow in this field. We believe our recommendations can help the readers perform more robust and sound pangenomic analyses and to generate more reliable results.

Excision of arteriovenous malformation in an emergency situation - A case report.

An arteriovenous malformation (AVM) is an infrequent congenital vascular anomaly that can affect the vasculature and involve the endothelium and neighboring cells of any anatomical structure. AVMs are characterized histologically by abnormal AV shunts with atypical interconnecting capillary beds. AVM can cause functional and esthetic issues like face asymmetry, pain, osteolytic changes, and unanticipated hemorrhage or squeeze and tear of the surrounding tissue without causing any symptoms. The literature search yielded limited case reports on AVMs in the facial region. Insufficient diagnosis, limited knowledge, and a lack of literature can lead to severe bleeding and potentially fatal hemorrhagic incidents following dental procedures like tooth extraction, surgery, puncture wounds, or blunt injuries in the affected area. In this manuscript, we report a case of AV malformation involving the left cheek and buccal mucosa region in a 37-year-old male patient who reported uncontrolled bleeding after trauma. This report highlights the management of AV malformation in an emergency by facial artery ligation and surgical excision.

Endoscopic clipping combined with cyanoacrylate injection vs. transjugular intrahepatic portosystemic shunt in the treatment of isolated gastric variceal bleeding: Randomized controlled trial.

OBJECTIVES: Although the incidence of isolated gastric varices type 1 (IGV1) bleeding is low, the condition is highly dangerous and associated with high mortality, making its treatment challenging. We aimed to compare the safety and efficacy of endoscopic clipping combined with cyanoacrylate injection (EC-CYA) vs. transjugular intrahepatic portosystemic shunt (TIPS) in treating IGV1. METHODS: In a single-center, randomized controlled trial, patients with IGV1 bleeding were randomly assigned to the EC-CYA group or TIPS group. The primary end-points were gastric variceal rebleeding rates and technical success. Secondary end-points included cumulative nonbleeding rates, mortality, and complications. RESULTS: A total of 156 patients between January 2019 and April 2023 were selected and randomly assigned to the EC-CYA group (n = 76) and TIPS group (n = 80). The technical success rate was 100% for both groups. The rebleeding rates were 14.5% in the EC-CYA group and 8.8% in the TIPS group, showing no significant difference (P = 0.263). Kaplan-Meier analysis revealed that the cumulative nonbleeding rates at 6, 12, 24, and 36 months for the two groups lacked statistical significance (P = 0.344). Similarly, cumulative survival rates at 12, 24, and 36 months for the two groups were not statistically significant (P = 0.916). The bleeding rates from other causes were 13.2% and 6.3% for the respective groups, showing no significant difference (P = 0.144). No instances of ectopic embolism were observed in either group. The incidence of hepatic encephalopathy (HE) in the TIPS group was statistically higher than that in the EC-CYA group (P = 0.001). CONCLUSION: Both groups are effective in controlling IGV1 bleeding. Notably, EC-CYA did not result in ectopic embolism, and the incidence of HE was lower than that observed with TIPS.

Double Tips for In-Plane Polarized Near-Field Microscopy and Spectroscopy.

Near-field optical microscopy and spectroscopy provide high-resolution imaging below the diffraction limit, crucial in physics, chemistry, and biology for studying molecules, nanoparticles, and viruses. These techniques use a sharp metallic tip of an atomic force microscope (AFM) to enhance incoming and scattered light by excited near-fields at the tip apex, leading to high sensitivity and a spatial resolution of a few nanometers. However, this restricts the near-field orientation to out-of-plane polarization, limiting optical polarization choices. We introduce double tips that offer in-plane polarization for enhanced imaging and spectroscopy. These double tips provide superior enhancement over single tips, although with a slightly lower spatial resolution (∼30 nm). They enable advanced studies of nanotubes, graphene defects, and transition metal dichalcogenides, benefiting from polarization control. The double tips allow varied polarization in tip-enhanced Raman scattering and selective excitation of transverse-electric and -magnetic polaritons, expanding the range of nanoscale samples that can be studied.

Transjugular antegrade transvenous obliteration, with and without portal decompression, for management of rectal variceal hemorrhage.

PURPOSE: To report antegrade transvenous obliteration, with or without concurrent portosystemic shunt creation, for the treatment of hemorrhagic rectal varices. MATERIALS AND METHODS: Eight patients, including five (62.5%) females and three (37.5%) males, with mean age of 55.8 ± 13.8 years (range: 30-70 years), underwent transjugular-approach antegrade transvenous obliteration of rectal varices, with or without portosystemic shunt creation. Demographic data, procedural details, technical success of variceal obliteration, clinical success, adverse events, and follow-up outcomes were retrospectively recorded. Clinical success was defined as resolution of rectal hemorrhage. RESULTS: Portal venous access was achieved via a transjugular intrahepatic approach in all patients. The inferior mesenteric vein was selected, and foamy sclerosant (1:2:3 mixture by volume of ethiodized oil: sodium tetradecyl sulfate: air) was injected into the rectal varices with antegrade balloon occlusion in seven (87.5%) and without balloon occlusion in one (12.5%). Five of eight (62.5%) patients underwent concomitant transjugular intrahepatic portosystemic shunt (TIPS) creation (mean diameter 8.4 ± 0.9-mm) immediately following transvenous obliteration. Technical success of variceal obliteration was achieved in all patients. There were no immediate post-procedural adverse events. There were no reported occurrences of rectal ischemia, perforation, or stricture following obliteration. Two (40%) of the patients who underwent concomitant TIPS creation developed hepatic encephalopathy within 30 days of the procedure, which was medically managed. Clinical resolution of hemorrhage was achieved in all patients with no recurrent rectal variceal hemorrhage during mean follow-up of 666 ± 396 days (range: 14 - 1,224 days). CONCLUSION: Transvenous obliteration, with or without concurrent TIPS creation, is feasible with promising results for the management of rectal variceal hemorrhage.

A Rare Case of Iatrogenic Urinary Bladder Injury During Inguinal Hernia Repair: Management Tips, Tricks and Pitfalls.

Iatrogenic injury of the bladder is a rare incidence during inguinal hernia repair in children, with serious consequences for such patients. Due to the scarcity of information on this matter, it is our goal to share our experience regarding the therapeutic approach to such a rare occurrence. Specifically, a 22-month-old male was admitted to our department with the complaint of vomiting, abdominal pain and anuria, two days after inguinal hernia repair. The child had distention of the inguinal hernia region and was lethargic. The diagnostic investigation did not reveal any significant findings. During surgical exploration, we discovered an injury to the bladder, while a large part of the dome of the bladder was ligated and subsequently became necrotic. After a reoperation and an enduring postoperative course, the patient finally recovered. Currently, the child is under observation. Therefore, it is of paramount importance for pediatric surgeons to be acquainted with the potential for bladder injury during inguinal hernia repair, ways to manage this complication, and various issues that may emerge during the therapeutic process.

A 27-Year-Old Female With JAK2 Mutation: A Case of Budd-Chiari Syndrome Secondary to Prolonged Oral Contraceptive Pill Use.

Individuals with Philadelphia chromosome-negative myeloproliferative neoplasms (MPNs) such as polycythemia vera and essential thrombocythemia (ET) demonstrate an increased thrombotic risk associated with JAK2 mutations. Physicians must take heed when treating these patients, to mitigate this pro-thrombotic state as much as possible. Failure to do so, or exacerbating the state, can lead to dire consequences. We present the case of a 27-year-old female with a history of ulcerative colitis (UC) and ET, currently taking estrogen-containing oral contraceptive pills (OCPs). She presented to the emergency department with rapid weight gain, jaundice, nausea, and diarrhea and was found to have obstructive jaundice and thrombotic burden that extended into the portal, mesenteric, splenic, and hepatic veins. On the second attempt, a successful transjugular intrahepatic portosystemic shunt procedure was performed, resulting in improved venous flow. This case underscores the importance of cautious medication use, especially OCPs, in patients with hypercoagulable states due to JAK2 mutations, for example, the V617F mutation in JAK2. It emphasizes the need for vigilant monitoring, individualized management, and a multidisciplinary approach to mitigate thrombotic complications. Increased awareness and continued research are crucial for optimizing treatment strategies for patients with MPNs and associated genetic mutations.

Tricks and tips for trips.

Finding out about sample preparation and transportation of structural biology samples in Acta Crystallographica F, Structural Biology Communications.

Discovering genes and microRNAs involved in human lung development unveils IGFBP3/miR-34a dynamics and their relevance for alveolar differentiation.

BACKGROUND: During pseudoglandular stage of the human lung development the primitive bronchial buds are initially conformed by simple tubules lined by endoderm-derived epithelium surrounded by mesenchyme, which will progressively branch into airways and start to form distal epithelial saculles. For first time alveolar type II (AT2) pneumocytes appears. This study aims to characterize the genes and microRNAs involved in this differentiation process and decipher its role in the starting alveolar differentiation. METHODS: Gene and microRNA profiling was performed in human embryonic lungs from 7 to 12 post conception weeks (pcw). Protein expression location of candidate genes were analyzed by immunofluorescense in embryonic lung tissue sections. mRNA/miRNA target pairs were identified using computational approaches and their expression was studied in purified epithelial/mesenchymal cell populations and in isolated tips and stalks from the bronchial tree. Additionally, silencing experiments in human embryonic lung mesenchymal cells and in human embryonic tip-derived lung organoids were performed, as well as organoid differentiation studies. AT2 cell markers were studied by qRT-PCR and by immunofluorescence. The TGFB-ß phosphorylated pathways was analyzed with membrane protein arrays. Lung explants were cultured in air/liquid interface with/without peptides. RESULTS: We identified 88 differentially expressed genes, including IGFBP3. Although IGFBP3 mRNA was detected in both epithelial and mesenchymal populations, the protein was restricted to the epithelium, indicating post-transcriptional regulation preventing IGFBP3 protein expression in the mesenchyme. MicroRNA profiling identified miR-34a as an IGFBP3 regulator. miR-34a was up-regulated in mesenchymal cells, and its silencing in human embryonic lung mesenchymal cells increased IGFBP3 levels. Additionally, IGFBP3 expression showed a marked downregulation from 7 to 12 pcw, suggesting its involvement in the differentiation process. The differentiation of human tip-derived lung embryonic organoids showed a drastic reduction in IGFBP3, supported by the scRNAseq data. IGFBP3 silencing in organoids activated an alveolar-like differentiation process characterized by stem cell markers downregulation and upregulation of AT2 markers. This process was mediated by TGFß signalling inhibition and BMP pathway activation. CONCLUSIONS: The IGFBP3/miR-34a axis restricts IGFBP3 expression in the embryonic undifferentiated lung epithelium, and the progressive downregulation of IGFBP3 during the pseudoglandular stage is required for alveolar differentiation.

Evaluation of clinical outcome and predictors of mortality in patients undergoing antegrade transvenous variceal embolisation in adjunct to salvage TIPS for active uncontrolled gastric variceal bleeding.

OBJECTIVES: Salvage TIPS is indicated in patients with active endoscopically uncontrollable variceal bleeding. TIPS alone is not effective in management of gastric varices and BRTO requires favourable variceal anatomy. Concomittant placement of a TIPS stent with antegrade variceal embolisation leads to control of gastric variceal bleeding with no significant increase in portal pressure. MATERIALS AND METHODS: A single centre retrospective observational study where patients with active uncontrollable gastric variceal bleeding were included in the study. Technical success of the procedure, 5-day rebleed, 6-weeks and 6-months survival as well as other additional outcomes were evaluated. RESULTS: A total of 18 patients were included in the study. Technical success was 100% and significant non-target embolisation was seen in 0% patients. 6 week and 6 month survival rates were 66.67% with an overall survival of 108.786 days (censored at 180 days). 5 day rebleed rate was 11.1%. A significant difference in CTP score (p = 0.03), MELD Na score (p = 0.022), requirement of intubation (p = 0.038), hemoglobin levels (p = 0.042), hematocrit value (p = 0.018), PRBC infusion required prior to and after the procedure (p = 0.045, 0.044) and presence of refractory shock (p = 0.013) was observed between the survival and the mortality group. Post variceal bleeding hemoglobin levels, MAP and MELD-Na scores were significant predictors of mortality. CONCLUSION: TIPS in adjunct to antegrade transvenous embolisation is a safe and effective modality for management of active uncontrolled gastric variceal bleeding in patients with variceal anatomy unfavourable for performing RTO.

Endoscopic Colorectal Stenting: Lessons Learned from a Two-Decade Journey.

Endoscopic colorectal stenting has gained momentum over the last two decades as a viable alternative to surgical intervention in a subgroup of patients with colorectal disease. Stenting can be used as a temporizing bridge to surgical intervention or as a definitive treatment measure. Patient selection and the technical expertise of the endoscopist are of paramount importance to optimize the clinical outcome. Technical skills in therapeutic endoscopy and the choice of proper equipment including the consumables are required for the conduct of a safe and successful procedure. In this article, we share the lessons learned from a two-decade journey of the senior author with therapeutic endoscopy.

Analysis of Microtubule Polymerization During Axon Outgrowth Using Fluorescently Labeled Microtubule Plus-End Tracking Proteins.

The study of microtubules arrangements and dynamics during axon outgrowth and pathfinding has gained scientific interest during the last decade, and numerous technical resources for its visualization and analysis have been implemented. In this chapter, we describe the cell culture protocols of embryonic cortical and retinal neurons, the methods for transfecting them with fluorescent reporters of microtubule polymerization, and the procedures for time-lapse imaging and quantification in order to study microtubule dynamics during axon morphogenesis.

Leaks after laparoscopic sleeve gastrectomy: 2024 update on risk factors.

PURPOSE: Leaks after sleeve gastrectomy remain a deadly complication significantly affecting outcomes and medical costs. The aim of the present review is to provide an updated decalogue on leak prevention. METHODS: Risk factors of leakage after LSG were examined based on an extensive review of literature (in period time 2016-2024) and summary of evidence was provided using Oxford levels of evidence scale. RESULTS: Pathogenesis of leakage after LSG still remain related to ischemic and mechanical factors and, therefore, no new evidence has been reported. Conversely, some technical aspect of the procedure has changed: bougie size, antrum resection, staple line reinforcement, and intraoperative leak testing. CONCLUSIONS: Bougie size 36 F is effective and safe achieving similar leakage rate compared to larger bougie sizes (EL:2) 2024 UPDATE; There is no significant difference in the leak rate between restrictive (< 6 cm) and conservative (6 cm) antrum resection (EL: 1) 2024 UPDATE; Surgical experience and case volume affect the leak rate more consistently than every kind of SLR (EL: 2) 2024 UPDATE; Intraoperative leak test after LSG represents a decision based on surgeon preference in absence of standardization (endoscopy, bubble test, methylene blue, indocyanine green.) and strong detection/prevention rate (EL: 3) 2024 UPDATE.

The bacterial displacement test: an in vitro microbiological test for the evaluation of intermittent catheters and urinary tract infection.

AIMS: Intermittent catheters (ICs) are commonly used in bladder management, but catheter-associated urinary tract infections (CAUTIs) remain challenging. Insertion tips may reduce the risk of CAUTIs by minimizing bacterial transfer along the urinary tract. However, there are few laboratory tests to evaluate such technologies. We describe the use of an adapted in vitro urethra agar model to assess bacterial displacement by ICs. METHODS AND RESULTS: Simulated urethra agar channels (UACs) were prepared with catheter-specific sized channels in selective media specific to the challenge organisms. UACs were inoculated with Escherichia coli and Enterococcus faecalis before insertion of ICs, and enumeration of UAC sections was performed following insertion. Four ICs were evaluated: Cure Catheter® Closed System (CCS), VaPro Plus Pocket™, Bard® Touchless® Plus, and SpeediCath® Flex Set. CCS demonstrated significantly reduced bacterial displacement along the UACs compared to the other ICs and was also the only IC with undetectable levels of bacteria toward the end of the UAC (representing the proximal urethra). CONCLUSION: The bacterial displacement test demonstrated significant differences in bacterial transfer between the test ICs with insertion tips, which may reflect their different designs. This method is useful for evaluating CAUTI prevention technology and may help guide future technology innovations.

Predictors of Duration of Transjugular Intrahepatic Portosystemic Shunt (TIPS) Procedure: A Retrospective Single-Center Study.

PURPOSE: To determine the relationship between clinical, procedural, hospital, and physician characteristics with the duration of the transjugular intrahepatic portosystemic shunt (TIPS) procedure. METHODS: This retrospective study included patients over 18 years of age who underwent an initial TIPS procedure between January 2005 and August 2020. Exclusion criteria were TIPS performed outside the institution and failed TIPS placement. A total of 154 records were included. Regression analyses were used to identify predictors of procedural duration. RESULTS: The mean age at TIPS placement was 57 years. Seventy percent of patients were male and non-Hispanic whites (80.5%). The mean duration of the TIPS procedure was 169 minutes (SD: 78). Procedural duration was shorter when the etiology of cirrhosis was viral (mean: 144 min, SD: 84, p=0.008); the reason for TIPS was ascites (152, SD: 66, p=0.01); and the procedure did not require additional access (153 min, SD: 67, p=<.0001). The main clinical predictor of procedural duration was baseline bilirubin (Beta coefficient (ß): 5.6 min, p=0.0007). In multivariable linear models, in those patients that did not require additional access, bilirubin (ß: 4.9 min, p=0.005), etiology of cirrhosis, and physician experience were the main predictors of TIPS procedure duration. The effect of baseline bilirubin on procedural duration increased in the ascites group (ß: 19.5 minutes, p=0.006), especially when additional access was not required. CONCLUSIONS:  The study demonstrates an association between baseline bilirubin, etiology of cirrhosis, and physician experience with the duration of the TIPS procedure. The mechanism underlying the positive association between baseline bilirubin and procedural time is possibly related to the degree of liver fibrosis.

EuroQol Toddler and Infant Populations (EQ-TIPS): Age-Related Differences in Performance.

OBJECTIVES: The EQ-TIPS was developed to measure the Health-Related Quality of Life in infants/toddlers. Considering the rapid development in this period, this study aimed to investigate age-related variations in EQ-TIPS performance. METHODS: Data from 551 infants/toddlers living with a health condition were analysed. Infants/toddlers were grouped by age: 0-6 months (n = 100), 6-12 months (n = 95), 12-24 months (n = 147), and 36-48 months (n = 97). Differences in item responses and item correlations across age groups were calculated by Kruskal-Wallis and Spearman's correlations, respectively. RESULTS: The report of problems was significantly higher for movement, play, and communication in the 36-48-month group compared to the 0-6-month group. There were strong correlations (r > 0.50) across all age groups between play and movement and communication and social interaction/play; neither pain nor eating showed a clear pattern of association. CONCLUSIONS: There is an age-related difference in the reporting of items linked to developmental milestones (movement, play, and communication) with most problems reported in the 36-48-month group when deviation from peers and continued dependence on caregivers is notable. Consideration should be given to including broader examples of play in the EQ-TIPS. Redefining the items to represent social communication and/or (social) emotion, rather than communication and social interaction, may be warranted. Future research should explore the psychometric performance of items to further inform item inclusion and/or revision.

Successful Treatment of Very Severe Sinusoidal Obstruction Syndrome After Gemtuzumab Ozogamicin With Transjugular Intrahepatic Portosystemic Shunt, Defibrotide, and High-Dose Corticosteroids: A Case Report.

Sinusoidal obstruction syndrome (SOS) is a rare but potentially life-threatening complication, usually described in the setting of hematopoietic stem cell transplantation (HSCT). The very severe forms have a high mortality rate (>80%) and need fast recognition and urgent treatment. In this case report, we describe a unique and successful treatment strategy. We present a 27-year-old patient with newly diagnosed CD33+ acute myeloid leukemia (AML). She was treated with induction chemotherapy (7+3 regimen) and gemtuzumab ozogamicin (GO). In the absence of other major risk factors, she developed a very severe SOS with multiple organ failure. She was successfully treated with the urgent insertion of a transjugular intrahepatic portosystemic shunt (TIPS), defibrotide, and high-dose corticosteroids. This case of successful treatment for very severe SOS supports a combination strategy involving the immediate mechanical reduction of portal hypertension through TIPS and drug-mediated inhibition of microvascular thrombosis. Furthermore, this case shows the need for an improved prevention strategy, including the identification of additional risk factors and biomarkers.

Psychometric validation of the Chinese versions of EQ-5D-Y-3L and the experimental EQ-TIPS in children and adolescents with COVID-19.

OBJECTIVES: Respiratory infectious diseases like COVID-19 profoundly impacts the health of children and adolescents, but validated instruments to measure their impacts on health-related quality of life (HRQoL) are lacking. The EQ-5D-Y-3L, widely used for youth HRQoL, now features a Chinese value set. The experimental EQ-TIPS addresses HRQoL assessment for toddlers and infants. This study tested the psychometric properties of both instruments in paediatric COVID-19 patients, and compared the performance of self-complete and proxy EQ-5D-Y-3L. METHODS: This longitudinal study recruited 861 COVID-19 patients aged 0-18 years and their parental caregivers, with 311 dyads completing the follow-up. Digital administration included the EQ-TIPS, the EQ-5D-Y-3L, and Overall Health Assessment (OHA). Controls comprised 231 healthy children. Analysis encompassed known-group validity, child-parent agreement, and responsiveness to change in disease severity and OHA. RESULTS: COVID-19 children exhibited lower HRQoL than non-infected peers. The EQ-TIPS and the EQ-5D-Y-3L distinguished groups by disease presence, severity and symptoms, showing moderate to good known-group validity (ESs: 0.45-1.39 for EQ-TIPS, 0.44-1.91 for self-complete EQ-5D-Y-3L, and 0.32-1.67 for proxy EQ-5D-Y-3L). Child-parent agreement was moderate to good for EQ-5D-Y-3L (ICC: 0.653-0.823; Gwet's AC1: 0.470-0.738), and responsiveness was good for both EQ-TIPS Level Sum Score (LSS) (ESs: 1.21-1.39) and EQ-5D-Y-3L index scores (ESs: 1.00-1.16). CONCLUSIONS: This study demonstrates the reliability, validity, and responsiveness of the experimental EQ-TIPS and the EQ-5D-Y-3L in paediatric COVID-19 patients. It is the first evidence of the EQ-TIPS' responsiveness, supporting its use in assessing the impact of COVID-19 on paediatric HRQoL.

Advancements in Interventional Radiology for Managing Hepatic Encephalopathy: A Comprehensive Review.

Hepatic encephalopathy (HE) is a complex neuropsychiatric syndrome resulting from liver dysfunction, leading to cognitive, behavioral, and motor impairments. The management of HE has traditionally relied on pharmacological treatments, dietary modifications, and liver transplantation. However, recent advancements in interventional radiology (IR) have introduced minimally invasive procedures that offer promising alternatives. This comprehensive review explores the latest IR techniques, including transjugular intrahepatic portosystemic shunt (TIPS), balloon-occluded retrograde transvenous obliteration (BRTO), portal vein embolization (PVE), and Yttrium-90 (Y90) radioembolization. The efficacy, clinical outcomes, and potential complications of these techniques are examined through an analysis of current studies and trials. The review highlights the benefits of IR in reducing portal hypertension and improving hepatic blood flow, ultimately alleviating HE symptoms. Additionally, it underscores the importance of multidisciplinary collaboration, ongoing research, and the development of clear patient selection criteria to optimize the use of IR in HE management. By integrating these advancements into clinical practice, healthcare providers can enhance the quality of care and improve outcomes for patients with HE.

The Long-Term Impact of the Tips From Former Smokers® Campaign on Calls to 1-800-QUIT-NOW, 2012-2023.

INTRODUCTION: There is substantial evidence that mass media campaigns increase calls to quitlines and smoking cessation. In 2012, the Centers for Disease Control and Prevention launched the first federally funded national tobacco education campaign, Tips From Former Smokers® (i.e. Tips), which has since aired television advertisements annually. To date, no studies have examined the long-term effect of a national smoking cessation campaign on quitline calls. This study examined the long-term impact of Tips television ads on calls to 1-800-QUIT-NOW from 2012 through 2023. METHODS: Exposure to the Tips campaign was measured using weekly television gross rating points (GRPs) in each U.S. designated market area. We obtained data on calls to 1-800-QUIT-NOW from the National Cancer Institute and used linear regression to model calls to 1-800-QUIT-NOW, from 2012 through 2023, as a function of weekly media market-level GRPs for Tips television ads. Using the regression model results, we calculated predicted values of calls to 1-800-QUIT-NOW across observed GRP values to determine the total additional calls to 1-800-QUIT-NOW that were attributable to the Tips campaign during 2012-2023. RESULTS: Tips GRPs were positively and significantly associated with calls to 1-800-QUIT-NOW across all years (b = 39.94, p < 0.001). Based on this association, we estimate the Tips campaign generated nearly 2.1 million additional calls to 1-800-QUIT-NOW during 2012-2023. CONCLUSIONS: Exposure to the Tips campaign has consistently and significantly increased calls to tobacco quitlines. IMPLICATIONS: Quitlines provide evidence-based support to help people quit smoking. They have been shown to increase the likelihood of successfully quitting. Mass media campaigns have promoted quitline services, and quitline calls have increased significantly with media promotion. The long-term effect of campaigns - like the Centers for Disease Control and Prevention's Tips From Former Smokers® (i.e. Tips) - on quitline calls has not been determined. From 2012 through 2023, exposure to the Tips campaign is estimated to have generated nearly 2.1 million additional calls to 1-800-QUIT-NOW. This study supports continued use of mass media to promote quitlines.