Deployment and transcriptional evaluation of nitisinone, an FDA-approved drug, to control bed bugs.

Bed bugs are blood-feeders that rapidly proliferate into large indoor infestations. Their bites can cause allergies, secondary infections and psychological stress, among other problems. Although several tactics for their management have been used, bed bugs continue to spread worldwide wherever humans reside. This is mainly due to human-mediated transport and their high resistance to several classes of insecticides. New treatment options with novel modes of action are required for their control. In this study, we evaluated the use of nitisinone (NTBC), an FDA-approved drug, for bed bug control in an insecticide-susceptible (HH) and an insecticide-resistant (CIN) population. Although NTBC was lethal to both populations when administered orally or applied topically in very low doses, we observed a slight but significant resistance in the CIN population. Transcriptomic analysis in both populations indicated that NTBC treatment elicited a broad suppression of genes associated with RNA post-transcriptional modifications, translation, endomembrane system, protein post-translational modifications and protein folding. The CIN population exhibited higher ATP production and xenobiotic detoxification. Feeding studies on a mouse model highlight that NTBC could be used as a control method of bed bugs by host treatment. The results demonstrate that NTBC can be used as a new active ingredient for bed bug control by topical or oral treatment and shed light on the molecular mechanisms of suppressed tyrosine metabolism following NTBC treatment.

Importance of blood pressure monitoring in the acute phase of stroke. An update.

INTRODUCTION: The evaluation of blood pressure (BP) is essential in the acute phase of stroke. Although ambulatory blood pressure monitoring (ABPM) is a validated method for BP control, there are few studies assessing the usefulness of ABPM in the acute phase of stroke. DEVELOPMENT: A systematic review was carried out according to the PRISMA criteria in the PubMed/Medline and Scopus databases. Those articles that analysed the use of ABPM in the first days after suffering a stroke from 1992 to 2022 were selected. Those articles focused on the post-acute or sequelae phase of the stroke, with a sample size of less than 20 and those where the primary objective was different from the defined one. A total of 28 articles were included. CONCLUSIONS: The use of ABPM in patients with recent stroke demonstrates that the normal circadian profile of BP is altered in more than two-thirds of patients and that this will be fundamentally conditioned by the haemodynamic changes that occur on autoregulation of cerebral blood flow, the type of stroke or the response to treatment. Furthermore, these changes in BP have prognostic implications and are correlated with functional status, stroke recurrence and mortality, among others. However, although they continue to be a growing area of research, new studies are needed to clarify the real role of this technique in patients with acute stroke.

Mortality and cardiovascular risk in vasculitis ANCA. Importance of hypertension and renal function. Experience from southern Spain.

BACKGROUND: Cardiovascular disease (CVD) is one of the principal causes of death in antineutrophil cytoplasmic antibody-(ANCA)-associated vasculitis (AAV). OBJECTIVES: To evaluate the mortality and it's causes and CVD and its vascular risk factors (VRFs) in AAV patients in Andalusia. METHODS: A multicenter cohort of 220 AAV patients followed-up from 1979 until June 2020 was studied in Andalussia, south of Spain. The information, including socio-demographic and clinical data was recorded retrospectively through chart review. Data was analysed using Chi2, ANOVA and Cox proportional hazards regresion as uni and multivariate test with a 95% confidence interval (CI). RESULTS: During a mean ± standard deviation follow-up of 96.79 ± 75.83 months, 51 patients died and 30 presented at least one CVE. Independent prognostic factors of mortality were age (HR 1.083, p=0.001) and baseline creatinine (HR 4.41, p=0.01). Independent prognostic factors of CVE were age [hazard ratio (HR) 1.042, p=0.005] and the presence of hypertension (HTN) six months after diagnosis (HR 4.641, p=0.01). HTN, diabetes and renal failure, all of these important VRFs, are more prevalent in AAV patients than it is described in matched general population. CONCLUSIONS: Age and baseline renal function, but not CVEs, are predictors of mortality and age and early HTN are independent predictors for having a CVE. CVD screening in AAV patients is demanded.

[Importance of hypertension in patients after the first stroke: retrospective cohort study]. / Importancia de la hipertensión en pacientes con primer episodio de ictus isquémico: estudio retrospectivo de 20 años.

BACKGROUND AND OBJECTIVES: Hypertension (HT) is a key risk factor for stroke. We evaluated whether the importance of hypertension on the risk of ischemic stroke has increased in recent decades. METHODS: Retrospective study of patients with ischemic stroke discharged from 3 hospitals in Seville (Spain), during the periods: 1999-2001, 2014-2016 and 2019-2020. RESULTS: 1,379 patients were included, 42.6% women, mean age 69.1 (±11) years. HT was the most prevalent vascular risk factor in all periods, with a progressive increase in hypertensive patients (65.9% vs 69.6% vs 74%; P=.029). HT was especially frequent in patients≥80 years (73% vs 81.9% vs 85.2%; P=.029). At discharge, progressively more antihypertensive drugs were used (in 65% vs 85.1 vs 90.2% of patients; P=.0001), with a clear increase in the number of antihypertensive drugs used (mean 0.9±0.8 vs 1.5±1 vs 1.8±0.8 drugs, P=.0001). The use of diuretics (13.7%-39.3%-65.3%; p=0.0001), ACE inhibitors (35.5%-43.3%-53.4%; P=.0001) and angiotensin receptor blockers (12.2%-24%-32.4%; P=.0001) increased progressively. On the contrary, the use of calcium antagonists decreased (24%-19.9%-13.7%; P=.0001). CONCLUSIONS: In the last 2 decades there has been a greater role for HT among patients with their first ischemic cerebrovascular event. Greater and better control of HT is necessary to reduce the enormous burden of cerebrovascular disease.

Pharmacological therapy in the secondary prevention of ischemic stroke in the oldest-old patients: has it improved in recent decades?

INTRODUCTION: Population aging has caused an increase in strokes in very elderly patients (VEP). We assess how secondary prevention of ischemic stroke has changed in VEP in recent decades. METHOD: Retrospective study of discharges due to ischemic stroke in the Virgen Macarena, Virgen del Rocio and Valme hospitals in Seville (Spain), during the periods 1999-2001, 2014-16 and 2019-2020. VEP were considered those with ≥80 years. RESULT: We studied 1806 patients, 349 (19.3%) were VEP. Over the years, VEPs have doubled (13.5% vs. 25.9% and 28% p = 0.0001) and age has increased (83.3 ±â€¯3 vs. 84.1 ±â€¯3 vs. 85.2 ±â€¯4 p = 0.001). Comparing the periods, the VEPs have more hypertension (69.9% vs. 84.8% vs. 84.6%; p = 0.0001) and dyslipidemia (12% vs. 41.7% vs. 52.3%; p = 0.0001) and have prescribed more antihypertensives (69.1% vs. 86.7% vs. 92.3%; p = 0.0001), statins (5.3% vs. 78% vs. 81.5%; p = 0.0001) and anticoagulants (16.5% vs. 19.4% vs. 53.1%; p = 0.001), increasing the number of antihypertensives (1 ±â€¯0.9 vs. 1.6 ±â€¯0, 9 vs. 1.9 ±â€¯0.8 drugs p = 0.0001), and high-intensity statins (2.3% vs. 42.7 vs. 69.2% p = 0.0001). Comparing the VEPs with the younger ones, there were no differences in antihypertensive treatment in any period, there were differences in antithrombotic treatment in the first period, and with statins the differences were maintained until the end. CONCLUSIONS: In the last 20 years the number of VEPs has doubled, exceeding a quarter of the discharges. Although there is improvement in secondary stroke prevention in VEPs, there is room for improvement.

Fracture Load of Molars Restored with Bulk-fill, Flowable Bulk-fill, and Conventional Resin Composite After Simulated Chewing.

This study aimed to compare the fracture toughness of molars with wide mesio-occlusal-distal (MOD) cavities restored with regular and flowable bulk-fill resin composite and a conventional resin composite after 250,000 mechanical cycles of chewing simulation. Thirty-two extracted mandibular third molars were selected and class II MOD cavities involving 2/3 of the intercuspal width and 4 mm depth were prepared. Teeth were divided into four groups based on resin composite type and insertion technique (n=8): (1) CT, unprepared teeth (control); (2) CV, conventional resin composite (Tetric N-Ceram, Ivoclar Vivadent) with incremental technique; (3) R-BF, regular bulk-fill resin composite (Tetric N-Ceram Bulk Fill, Ivoclar Vivadent) with a single increment; and (4) F-BF, flowable bulk-fill resin composite (Tetric N-Flow Bulk Fill, Ivoclar Vivadent) with a single increment, except for a 1-mm-thick layer at the occlusal surface, restored with conventional resin composite (Tetric N-Ceram). All specimens were evaluated to detect the presence and propagation of enamel cracks using a LED transilluminator before and after 250,000 mechanical cycles (SD Mechatronic GmbH). After a chewing simulation, they were subjected to a compressive force in a universal testing machine (DL-2000, EMIC) until fracture. The maximum fracture load of the specimens was measured (N) and the fracture patterns were classified based on the fracture site (above or below the cementoenamel junction [CEJ]). Data were statistically analyzed with one-way ANOVA. All specimens survived after 250,000 mechanical cycles, and no statistically significant differences among groups were observed regarding the fracture toughness (p<0.05). The fracture analysis demonstrated that failures below the CEJ were more common in CV (75%), while CT, R-BF, and F-BF showed this type of failure in 38%, 63%, and 63% of the specimens, respectively. The results of the crack analysis showed that the occurrence of new cracks and crack propagation was also higher in CV (33.3%), followed by R-BF, F-BF, and CT (14%, 14%, and 11% of the specimens, respectively). Teeth restored with regular and flowable bulk-fill composites showed similar fracture toughness after the chewing simulation compared to those restored with the conventional resin composite and unprepared teeth. Furthermore, teeth restored with both regular and flowable bulk-fill composites showed a lower incidence of enamel cracks and fractures below the CEJ compared to those restored with the conventional resin composite.

Addressing high cervical cancer rates in the Rio Grande Valley along the Texas-Mexico border: a community-based initiative focused on education, patient navigation, and medical provider training/telementoring.

AIMS: Cervical cancer incidence and mortality rates are approximately 55% higher in the Rio Grande Valley (RGV) along the Texas-Mexico border compared with the average rates in the US. Our aim was to improve cervical cancer prevention efforts in the RGV through a comprehensive multilevel intervention initiative focused on community education, patient navigation, and training of local providers. METHODS: We initiated a program in the RGV which consisted of (1) community education, (2) patient navigation, and (3) a training/mentoring program for local medical providers including hands-on training courses coupled with telementoring using Project ECHO® (Extension for Community Health Outcomes). We assessed the number of women undergoing cervical cancer screening, diagnosis, and treatment at three participating clinics caring for underserved women in the region. RESULTS: From November 2014 to October 2018, 14,846 women underwent cervical cancer screening. A total of 2030 (13.7%) women underwent colposcopy for abnormal results (179% increase over baseline) and 453 women underwent loop electrosurgical excision procedures (LEEPs) for treatment of cervical dysplasia. Invasive cancer was diagnosed in 39 women who were navigated to a gynecologic oncologist for treatment. Seven local medical providers were trained to perform colposcopy and/or LEEP. Project ECHO telementoring videoconferences were held every 2 weeks for a total 101 sessions with an average of 22 participants per session and a total of 180 patient cases presented and discussed. CONCLUSIONS: Our program led to a large number of women undergoing diagnosis and treatment of cervical dysplasia in the RGV. If sustained, we anticipate these efforts will decrease cervical cancer rates in the region. The program is currently being expanded to additional underserved areas of Texas and globally to low- and middle-income countries.

Trends in the use of statins after ischaemic stroke: Have clinical practices changed?

INTRODUCTION: The role of statins after ischaemic stroke changed with the publication of the SPARCL study in 2006. We analyse how this has influenced the prescription of statins in this patient population. METHODS: We conducted a retrospective study of patients discharged with ischaemic stroke at the Virgen Macarena, Virgen del Rocío, and Valme hospitals in Seville (Spain) over two periods: 1999-2001 and 2014-2016. RESULTS: The study included 1575 patients: 661 (42%) were women and mean age (standard deviation) was 69 (10) years. Patients from the later period are older (68 [10] vs 71 [11]; P = .0001); include a higher proportion of women; and present higher rates of dyslipidaemia, hypertension, and diabetes. At discharge, statins were used in 18.7% of patients (vs 86.9% in the first period; P = .0001), with high-intensity statins prescribed in 11.1% of cases (vs 54.4%; P = .0001). In both periods, atorvastatin was the most commonly prescribed statin (80 mg: 6% vs 42.7%; 40 mg: 5.1% vs 11.1%). In the first period, the use of statins and high-intensity statins was correlated with hypercholesterolaemia, and inversely correlated with age. In the second period, statin use was correlated with hypertension and hypercholesterolaemia, and high-intensity statin use was correlated with ischaemic heart disease and inversely correlated with age. CONCLUSION: There has been a clear change in the prescription of statins to patients with ischaemic stroke at discharge. However, many patients remain undertreated and the use of these drugs needs to be optimised.

Antinuclear antibodies detected by Enzyme-Linked Immunosorbent Assay (ELISA) in severe COVID-19: clinical and laboratory associations.

OBJECTIVE: Antinuclear antibodies (ANA) are detected in approximately a quarter of COVID-19 patients when assessed by indirect immunofluorescence. Since there is no information, our study investigated the presence of ANA detected by Enzyme-Linked Immunosorbent Assay (ELISA) and its clinical and laboratory associations. PATIENTS AND METHODS: A longitudinal study was conducted on 92 patients with severe COVID-19, 20 patients with acute myocardial infarction, and 25 healthy subjects. Blood samples were obtained at hospital admission. Commercial ELISA was used to detect ANA, while flow cytometry was used to measure serum interferons. RESULTS: ANAs were positive in 8.6% of COVID-19 patients, 10% of myocardial infarction patients, and 4% in healthy individuals (p=0.676). COVID-19 patients with ANA+ had less ferritin, troponin, and neutrophils but more albumin and lymphocytes than ANA- patients. Serum levels of type I, II, and III interferons were similar between groups. At follow-up, all ANA+ patients survived, while mortality was significant in ANA- patients (0 vs. 36%; p=0.048). CONCLUSIONS: ANA detection is not increased in severe cases of COVID-19 when assessed by ELISA. However, its presence appears to be associated with a less aggressive disease phenotype, regardless of circulating levels of interferons.

Assessment of Noninvasive Markers of Steatosis and Liver Fibrosis in Human Immunodeficiency Virus-Monoinfected Patients on Stable Antiretroviral Regimens.

Background: Nonalcoholic fatty liver disease (NAFLD) is a major nonacquired immune deficiency syndrome-defining condition for persons with human immunodeficiency virus (PWH). We aimed to validate noninvasive tests for the diagnosis of NAFLD in PWH. Methods: This is a cross-sectional study of PWH on stable antiretroviral therapy with persistently elevated transaminases and no known liver disease. The area under the receiver operating characteristic curve (AUROC) was calculated to compare the diagnostic accuracy of liver biopsy with abdominal ultrasound, transient elastography (TE) (including controlled attenuation parameter [CAP]), and noninvasive markers of steatosis (triglyceride and glucose index [TyG], hepatic steatosis index [HSI], fatty liver index [FLI]) and fibrosis ([FIB]-4, aminotransferase-to-platelet ratio index [APRI], NAFLD fibrosis score). We developed a diagnostic algorithm with serial combinations of markers. Results: Of 146 patients with increased transaminase levels, 69 underwent liver biopsy (90% steatosis, 61% steatohepatitis, and 4% F ≥3). The AUROC for steatosis was as follows: ultrasound, 0.90 (0.75-1); CAP, 0.94 (0.88-1); FLI, 0.81 (0.58-1); HSI, 0.74 (0.62-0.87); and TyG, 0.75 (0.49-1). For liver fibrosis ≥F3, the AUROC for TE, APRI, FIB-4, and NAFLD fibrosis score was 0.92 (0.82-1), 0.96 (0.90-1), 0.97 (0.93-1), and 0.85 (0.68-1). Optimal diagnostic performance for liver steatosis was for 2 noninvasive combined models of tests with TyG and FLI/HSI as the first tests and ultrasound or CAP as the second tests: AUROC = 0.99 (0.97-1, P < .001) and 0.92 (0.77-1, P < .001). Conclusions: Ultrasound and CAP performed best in diagnosing liver steatosis, and FLI, TyG, and HSI performed well. We propose an easy-to-implement algorithm with TyG or FLI as the first test and ultrasound or CAP as the second test to accurately diagnose or exclude NAFLD.

Predictive factors for mortality and intensive care unit admission in patients with emphysematous pyelonephritis: 5-year experience in a tertiary care hospital.

INTRODUCTION AND OBJECTIVES: Emphysematous pyelonephritis is a life-threatening infection of the kidney and surrounding tissues associated with a high mortality rate. The aim of this study was to determine predictive factors for mortality and intensive care unit admission in patients with emphysematous pyelonephritis, and to propose a therapeutic algorithm based on current literature and our experience. METHODS: A retrospective study was done including patients with emphysematous pyelonephritis in a single center in the north of Mexico from 2011 to 2016. Demographic, clinical, microbiological and biochemical parameters, therapeutic management, and outcomes were assessed. Factors associated with admission to intensive care unit and mortality were determined. Comparison was assessed using X2 test for categorical variables, and T-test for numerical variables. Univariate and multivariate logistic regression analyses were performed. Statistical significance was set at P < .05. RESULTS: A total of 63 patients were included, of which 55 (87.3%) were females, with a mean age of 55.5 ±â€¯12.2 years. The most common comorbidities were diabetes and hypertension. Escherichia coli was the most common isolated microorganism (51.7%) and extended-spectrum beta-lactamase-producing agents were reported in 31.7%. Conservative therapy was provided to 38.7%, double J stent 42.9%, open/percutaneous drainage 12.7%, and nephrectomy 25.3%. Overall mortality and intensive care admission were 20.6% and 36.5%, respectively. In the multivariate analysis, hemodynamic instability (P = .005), qSOFA ≥ 2 (P = .003), hypoalbuminemia (P = .02), and early nephrectomy (P = .002) were associated with intensive care admission. Huang scale 4 (P = .006) and early nephrectomy (P = .001) were associated to mortality. CONCLUSIONS: Emphysematous pyelonephritis is a life-threatening disease and evidence of management is based in small case series due to the low incidence of this condition. Hemodynamic instability, hypoalbuminemia, qSOFA ≥ 2, Huang scale ≥3, and early nephrectomy are associated with poor prognosis.

[Morning blood pressure surge in patients with recent ischaemic stroke: the neglected factor]. / Incremento matutino de la presión arterial en pacientes con ictus isquémico reciente: el factor olvidado.

INTRODUCTION: High morning BP surge (MBPS) has been associated with an increased risk of cardiovascular events. We evaluated the presence of a high MBPS in patients with recent ischaemic stroke. MATERIAL AND METHODS: A case-control study was carried out. One hundred patients with an ischaemic stroke in the previous 6 months and fifty hypertensive patients without cardiovascular disease were included as controls. RESULTS: 61 lacunar (LAC) and 39 non-lacunar (NLAC) strokes were studied. The mean age was 65±11 years, and 60 (40%) patients were women. High MBPS was present in 9% of strokes (in 5 LAC and 4 NLAC) and in 8% of controls (p not significant [NS]), with a similar mean value of MBPS in both groups: 23.9±14mmHg and 24.9±15mmHg respectively (p=NS), although the control patients had a higher office BP (systolic [p=.008] and diastolic [p=.0001]), 24h systolic BP (p=.028) and daytime systolic BP (p=.022). Among the stroke patients, high MBPS was associated with previous coronary heart disease (p=.005), circadian BP pattern (p=.029), but not with the type of antihypertensive treatment prescribed. In multivariate analysis, elevated MBPS was only associated with previous coronary artery disease (p=.001). CONCLUSIONS: Approximately one in ten patients with recent ischaemic stroke has a high MBPS. Strategies to detect and treat high MBPS after a stroke are needed.

Resistant hypertension and Morgagni hernia: The importance of nocturnal hypoxaemia.

A 51-year-old woman consulted for resistant arterial hypertension despite adequate antihypertensive treatment. Physical examination and analytical study showed no relevant abnormalities, with pulse oximeter saturation of 95%. The study highlighted nocturnal respiratory polygraphy with data of mild intensity sleep apnoea syndrome, and severe nocturnal hypoxaemia (apnoea hypopnoea index per hours of sleep [AHI] 7.8; desaturation index per hour [ODI]: 12.6. Oxygen-medium saturation: 89%, minimum saturation: 72%. CT90: 34.2%). The chest X-ray showed elevation of the right hemidiaphragm, and the chest computed tomography (CT) revealed a Morgagni hernia with a maximum diameter of 20cm. After adjusting the antihypertensive treatment, the patient was referred to General Surgery for intervention. The onset of resistant hypertension secondary to severe nocturnal hypoxemia from a large Morgagni hernia has not been previously described in the literature.

Water dynamics over a Western Patagonian watershed: Land surface changes and human factors.

Warming trends in Patagonia and severe droughts in recent decades are still poorly understood in terms of their hydrological effects. The effects of climate change on water dynamics in addition to human water management could generate a future water scarcity scenario in one of the regions with the most abundant water resources of Chile. The aim of this work is to focus on assessing the impacts of warming trends on water dynamics in the Patagonian Simpson River watershed during the last two decades. We estimated anomalies in the main components of water balance such as precipitation (P), snow cover (SC), evapotranspiration (ET) and streamflows (Q) as well as surface variables and meteorological forcing (i.e. air temperature - Ta, solar radiation - RS, land surface temperature - LST). The processed data were obtained from remote sensing, reanalysis and in-situ data. We implemented a trend analysis for each variable in the period 2000-2019 at monthly, seasonal and annual scale. Results showed a warming trend in Ta and LST of about 1.2 °C and 2.1 °C, respectively, concentrated mainly in the autumn and winter seasons. Although P showed non-significant trends, Q diminished significantly at rates of more than 9.1 m3/s/decade, representing 36% of its historical mean. However, the decreases in Q are seen only in the maximum (spring) and minimum (summer) seasonal flows. These decreases are explained by significant increases in ET, led by a positive feedback of its drivers (LST, Ta and RS), which is directly linked to the impact of warming and an associated vegetation greenness in the watershed, as well as a decrease in SC during winter that feeds the Simpson River during spring and summer. The decrease in Q is reinforced by the intensification of water withdrawals in recent decades, as shown by an accelerated increase in water rights for agricultural and drinking uses. In a context of water scarcity and increasing and extreme droughts, this work contributes to further understanding water dynamics in western Patagonia, providing support for policy and decision-making when defining sustainable productive practices at watershed scale.

[Hyperlipoproteinaemia(a) and stroke. A case report of a family with early-onset severe atherosclerotic disease]. / Hiperlipoproteinemia(a) e ictus. A propósito de una familia con enfermedad ateroesclerótica grave precoz.

TITLE: Hiperlipoproteinemia(a) e ictus. A propósito de una familia con enfermedad ateroesclerótica grave precoz.

New ABO intron 1 variant alleles.

Unusual and discrepant ABO phenotypes are often due to genetic variants that lead to altered levels or activity of ABO transferases and consequently to altered expression of ABO antigens. This report describes eight genetic alterations found in 15 cases with reduced or undetectable expression of ABO antigens. Forward and reverse ABO grouping was performed by standard gel or tube methods. Adsorption-heat elution and saliva testing for H and A substances followed the AABB technical manual procedures. Genomic DNA extracted from whole blood was PCR-amplified to cover the entire ABO coding sequence, splice junctions, proximal promoter, and intron 1 enhancer. Amplification products were sequenced by next-generation or Sanger dideoxy methods, either directly or after cloning into a bacterial plasmid vector. Eight unreported alleles were found in the 15 cases analyzed. Alleles ABO*A(28+1C) and ABO*A(29-5G) harbor variants that alter the consensus sequence at the intron 1 donor and acceptor splice sites, respectively. The other alleles harbor variants that alter the consensus sequence at transcription factor-binding sites in the intron 1 enhancer: specifically, ABO*A(28+5792T), ABO*A(28+5859A), and ABO*A(28+5860G) at GATA-1 sites; ABO*B(28+5877T) and ABO*B(28+5878G) at a RUNX1 site; and ABO*A(28+5843A) at or near a C/EBP site. Molecular and serologic characterization of ABO alleles can help in their future identification and in the resolution of discrepancies.Unusual and discrepant ABO phenotypes are often due to genetic variants that lead to altered levels or activity of ABO transferases and consequently to altered expression of ABO antigens. This report describes eight genetic alterations found in 15 cases with reduced or undetectable expression of ABO antigens. Forward and reverse ABO grouping was performed by standard gel or tube methods. Adsorption-heat elution and saliva testing for H and A substances followed the AABB technical manual procedures. Genomic DNA extracted from whole blood was PCR-amplified to cover the entire ABO coding sequence, splice junctions, proximal promoter, and intron 1 enhancer. Amplification products were sequenced by next-generation or Sanger dideoxy methods, either directly or after cloning into a bacterial plasmid vector. Eight unreported alleles were found in the 15 cases analyzed. Alleles ABO*A(28+1C) and ABO*A(29­5G) harbor variants that alter the consensus sequence at the intron 1 donor and acceptor splice sites, respectively. The other alleles harbor variants that alter the consensus sequence at transcription factor­binding sites in the intron 1 enhancer: specifically, ABO*A(28+5792T), ABO*A(28+5859A), and ABO*A(28+5860G) at GATA-1 sites; ABO*B(28+5877T) and ABO*B(28+5878G) at a RUNX1 site; and ABO*A(28+5843A) at or near a C/EBP site. Molecular and serologic characterization of ABO alleles can help in their future identification and in the resolution of discrepancies.