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BackgroundThe Cambodian National Influenza Center (NIC) monitored and characterized circulating influenza strains from 2009 to 2011.Methodology/principal findingsSentinel and study sites collected nasopharyngeal specimens for diagnostic detection, virus isolation, antigenic characterization, sequencing and antiviral susceptibility analysis from patients who fulfilled case definitions for influenza-like illness, acute lower respiratory infections and event-based surveillance. Each year in Cambodia, influenza viruses were detected mainly from June to November, during the rainy season. Antigenic analysis show that A/H1N1pdm09 isolates belonged to the A/California/7/2009-like group. Circulating A/H3N2 strains were A/Brisbane/10/2007-like in 2009 before drifting to A/Perth/16/2009-like in 2010 and 2011. The Cambodian influenza B isolates from 2009 to 2011 all belonged to the B/Victoria lineage represented by the vaccine strains B/Brisbane/60/2008 and B/Malaysia/2506/2004. Sequences of the M2 gene obtained from representative 2009-2011 A/H3N2 and A/H1N1pdm09 strains all contained the S31N mutation associated with adamantanes resistance except for one A/H1N1pdm09 strain isolated in 2011 that lacked this mutation. No reduction in the susceptibility to neuraminidase inhibitors was observed among the influenza viruses circulating from 2009 to 2011. Phylogenetic analysis revealed that A/H3N2 strains clustered each year to a distinct group while most A/H1N1pdm09 isolates belonged to the S203T clade.Conclusions/significanceIn Cambodia, from 2009 to 2011, influenza activity occurred throughout the year with peak seasonality during the rainy season from June to November. Seasonal influenza epidemics were due to multiple genetically distinct viruses, even though all of the isolates were antigenically similar to the reference vaccine strains. The drug susceptibility profile of Cambodian influenza strains revealed that neuraminidase inhibitors would be the drug of choice for influenza treatment and chemoprophylaxis in Cambodia, as adamantanes are no longer expected to be effective.

Background: Binaural hearing enables better speech comprehension in noisy environments and is necessary for acoustic spatial orientation. This study investigates speech discrimination in noise with separated signal sources and measures sound localization. The aim was to study characteristics and reproducibility of two selected measurement techniques which seem to be suitable for description of the aforementioned aspects of binaural hearing., Materials and Methods: Speech reception thresholds (SRT) in noise and test-retest reliability were collected from 55 normal-hearing adults for a spatial setup of loudspeakers with angles of ± 45° and ± 90° using the Oldenburg sentence test. The investigations of sound localization were conducted in a semicircle and fullcircle setup (7 and 12 equidistant loudspeakers)., Results: SRT (S -45 N 45 : -14.1 dB SNR; S 45 N -45 : -16.4 dB SNR; S 0 N 90 : -13.1 dB SNR; S 0 N -90 : -13.4 dB SNR) and test-retest reliability (4 to 6 dB SNR) were collected for speech intelligibility in noise with separated signals. The procedural learning effect for this setup could only be mitigated with 120 training sentences. Significantly smaller SRT values, resulting in better speech discrimination, were found for the test situation of the right compared to the left ear. RMS values could be gathered for sound localization in the semicircle (1,9°) as well as in the fullcircle setup (11,1°). Better results were obtained in the retest of the fullcircle setup., Conclusion: When using the Oldenburg sentence test in noise with spatially separated signals, it is mandatory to perform a training session of 120 sentences in order to minimize the procedural learning effect. Ear-specific SRT values for speech discrimination in noise with separated signal sources are required, which is probably due to the right-ear advantage. A training is recommended for sound localization in the fullcircle setup., (© 2024. The Author(s).)

Introduction: In spring 2020, the first nationwide lockdown in response to the spreading COVID-19 pandemic came into effect in Germany. From March to May, gambling venues, casinos, and betting offices were forced to close. This study explores how land-based gamblers respond to short-term closures of higher-risk forms of gambling. Which gamblers are particularly susceptible to switching to online gambling? Which are more likely to use the lockdown as an opportunity to quit or pause gambling? Potential parameters for these switching or cessation processes are identified using multivariate multinomial logistic regression analysis., Methods: The research questions are analyzed on the basis of quantitative data. For this purpose, a survey was conducted among members of a comparatively large German online access panel ("PAYBACK panel"). The sample of analysis consisted of 612 gamblers who had participated in at least one higher-risk form of gambling and had done so exclusively offline before the first lockdown (January and February 2020)., Results: A total of 37.1% of sports bettors ceased participation in higher-risk forms of gambling, compared to 64.1% of casino gamblers. Switching to online gambling, on the other hand, was a rather rare event, regardless of the form of gambling: the proportions differ between 7.7% (slot machines) and 10.9% (sports betting). In the multivariate model female gender, younger age, and a lower frequency of gambling before the first lockdown were found to be significant factors for quitting (instead of continuing) higher-risk offline gambling forms. Secondly, the analysis revealed that individuals with more pronounced cognitive distortions had an increased risk of switching to online gambling rather than staying offline., Discussion: A key finding of this study is that the temporary closure of offline venues does not result in a significant shift towards the online market. Instead, the results of this study show that these short, temporary closures of gambling venues were an appropriate opportunity to give individual groups of gamblers the opportunity to reflect, reduce or quit gambling. It is worth considering implementing such temporary closures as a preventive measure in the future - this should be investigated in advance in further evaluation studies., Competing Interests: All authors have received grants for gambling research from the Federal Ministry of Health, several federal states and lottery providers., (Copyright © 2022 Kalke, Schütze, Lahusen and Buth.)

Introduction: Due to their euphoric and sedative effects, opioid analgesics have high potential for abuse and dependence. In the last decade in the USA and many Western European Countries the prescription rates of opioid analgesics have steadily increased., Objective: This study describes 5-year trends in the prescription of opioid analgesics and risk indicators such as duration, dose and 'doctor shopping' in Northern Germany., Methods: The annual rates of opioid analgesic prescriptions have been analysed for patients with statutory health insurance processed by the North German Pharmacy Data Center for the years 2011-2015. A distinction was made between non-cancer patients and cancer patients, and different groups according to prescription duration and dose level., Results: Between 2011 and 2015, the annual number of patients with opioid analgesic prescriptions increased from 500,000 to 550,000. About half of non-cancer-patients (85% of the total sample) and cancer patients received opioid analgesics for 90 days or less. The rates for long-term prescriptions (⩾9 months) ranged between 6-7% for non-cancer patients and 7-8% for cancer patients. Between 1.2-1.8% received opioid analgesics in doses of more than two defined daily doses. The majority of non-cancer patients with opioid analgesic prescriptions were female. The average age of non-cancer patients was 66 years. About 80% of non-cancer patients with first opioid analgesic prescription received World Health Organization step II medication., Conclusion: For the first time, this study provides comprehensive patient-related analyses of opioid analgesic prescriptions in Germany over a 5-year period. Despite a slight increase in the overall number of opioid analgesic prescriptions, an epidemic spread of opioid analgesics cannot be observed.

Aims: To examine the effectiveness of a one-day skills training program for increasing trauma inquiry in routine substance use disorder treatment., Design: Cluster-randomized two-armed controlled trial, with 12 substance use disorder (SUD) organizations operating 25 counseling centers, randomly assigned to training in trauma inquiry (13 counseling centers of 8 SUD organizations) or no training (12 counseling centers of 4 SUD organizations)., Setting: SUD counseling centers in Northern Germany., Cases: N = 5204 SUD counseling services., Intervention: The professionals assigned to the intervention group received a one-day training in trauma inquiry plus a 1.5-hour refresher session 3 months later. Professionals in the control group received no training., Measures: Over a 12-month period, professionals documented for each counseling service whether they asked the client about four traumatic events: physical abuse, emotional abuse, sexual abuse and neglect., Analysis: Primary outcomes were rates of asking about physical abuse, sexual abuse, emotional abuse and neglect in the 6 months after training. These were compared across conditions, while adjusting for baseline probabilities in the 6 months before training, using mixed-effects logistic regression., Findings: In the 6 months after training, the rate of asking about physical abuse was 18% higher in the SUD counseling services of trained professionals, relative to services of untrained professionals (OR = 1.18, 95% CI = [1.01-1.37, p = .035]). No effect was found for asking about sexual abuse, emotional abuse and neglect., Conclusion: A one-day training program in trauma inquiry, combined with a brief refresher session, was effective in increasing inquiries about physical abuse in routine counseling practice. The training was ineffective in increasing inquiries about sexual abuse, emotional abuse and neglect. The effectiveness of a one-day training of trauma inquiry might be increased by a longer training, or by combining it with additional elements, such as ongoing supervision., (Copyright © 2019. Published by Elsevier Inc.)

Objective: The majority of medication-dependent persons uses sedatives and hypnotics for many years. In this study we describe trends of benzodiazepine and z-drug prescriptions over a 10-years period., Methods: Prescriptions from 2006 to 2015 (based on German statutory health insurances) from the North German Pharmacy Computing Centre (NARZ) were analysed for the Federal states Schleswig-Holstein, Hamburg, Lower Saxony and Bremen. Data were classified as appropriate (in accordance with the guideline) or inappropriate/risky prescription patterns., Results: In a 10-years period 1.64 Million patients received benzodiazepine and/or z-drug prescriptions. Two thirds were women (65.3 %). More than half were 60 years or older. The percentage of patients with appropriate prescriptions (less than 2 months) increased from 51.7 % in 2006 to 60.2 % in 2015. The rate of inappropriate or risky prescription patterns which can be indicative of misuse or dependence dropped from 34.8 % in 2006 to 27.1 % in 2015., Conclusion: Over the past years the amount of inappropriate or risky benzodiazepine and z-drug prescriptions (which are not in accordance with the guideline) among patients with statutory health insurance is steadily decreasing. However, two fifths of the patients still received prescriptions for long-term intake or in higher doses. It remains unclear to what extent private prescriptions are used for inappropriate or long-term prescriptions of benzodiazepines and z-drugs., Competing Interests: Uwe Verthein erhielt innerhalb der letzten 3 Jahre Vortrags- und Beratungshonorar von Mundipharma GmbH sowie Reisekostenerstattungen von Mundipharma GmbH und CAMURUS GmbH. Die anderen Autoren hatten keine Verbindungen i. o. g. Sinne., (© Georg Thieme Verlag KG Stuttgart · New York.)

Background: An estimated 1.4 to 2.6 million people in German suffer from drug dependence. Most of them are long-term users of benzodiazepines (BZD), Z drugs (ZD), or opioid analgesics (OA)., Methods: This analysis is based on prescription data from patients of the national statutory health insurance system in the German federal states of Schleswig-Holstein, Hamburg, Bremen, and Lower Saxony. Drug-taking trends, duration, dosage, and long-term use of BZD, ZD, and OA in the years 2006 to 2015 are analyzed; prevalences are estimated for the years 2006 to 2016., Results: In 2006, 7.7% of patients received at least one prescription for a BZD, ZD, or OA; in 2016, 7.0% did. Over the period of analysis, a marked drop was seen in prescriptions of BZD and a slight fall in prescriptions of ZD (2006: BZD 3.5%, ZS 1.1%; 2016: BZD 2.0%, ZS 0.8%), but there was also an increase in prescriptions of OA, from 4.2% to 4.9%. The number of defined daily doses (DDD) prescribed per year fell for both BZD and ZD. For OA, the number of DDD prescribed per year rose from 2006 to 2009 and decreased by a small amount in subsequent years. The proportions of BZD and ZD patients who had long-term prescriptions fell over time, while the corresponding percentage of OA patients rose., Conclusion: Nearly one-fifth of all prescriptions for BZD were long-term prescriptions for an entire year, in violation of the relevant guidelines. The rising prevalence of OA use was in the expected range in view of the aging population, but the number of prescriptions rose among younger patients as well. This trend toward more common treatment with opioid analgesics should be critically examined.

Background: Despite the high rate of traumatic events in clients with substance use disorders, trauma exposure often remains undetected in a majority of treatment-seeking clients. Improving health professionals' knowledge and skills in the inquiry of traumatic events is therefore of utmost importance for appropriately addressing trauma-related treatment needs. However, professionals in substance use disorder treatment settings frequently report barriers to the inquiry about traumatic events, e.g., the fear of offending or harming the client. Such barriers should be addressed by trainings that aim to improve the systematic inquiry of traumatic events., Methods: Using a cluster-randomized trial, we examined whether a one-day training in trauma inquiry ('Learning How to Ask') would reduce professionals' perceived barriers to trauma inquiry. One hundred forty-eight professionals working in outpatient substance use disorder treatment centers were randomized to an intervention (n = 72) or a control group (n = 76). The professionals in the intervention group received a one-day training plus a refresher session 3 months later, the professionals in the control group received no training. At baseline, and at 3-month and 6-month follow-up, professionals rated on a four-point Likert scale regarding how strongly they agreed with statements about six common barriers to trauma inquiry, namely 'Feeling uncomfortable when asking about traumatic events', 'Fear of offending the client', 'Fear of retraumatizing the client', 'Fear that client may terminate treatment', 'Unsure whether authorities have to be informed when perpetrator is known', and 'No trauma-specific treatment available in my local area'., Results: The trained group experienced significant greater decreases in five of the six perceived barriers to the inquiry of traumatic events from baseline to 6-month follow-up than the control group ('Feeling uncomfortable when asking about traumatic events': b = - 0.32, 95% CI [- 0.52, - 0.12]; 'Fear of offending the client': b = - 0.33, 95% CI [- 0.56, - 0.09]); 'Fear of retraumatizing the client': b = - 0.45, 95% CI [- 0.69, - 0.22]; 'Fear that client may terminate treatment': b = - 0.28, 95% CI [- 0.49, 0.07]; 'No trauma-specific treatment available in my local area': b = - 0.25, 95% CI [- 0.51, - 0.01])., Conclusions: Our findings provide first evidence that a one-day training in trauma inquiry is effective in reducing common barriers to trauma inquiry, which may in turn improve detection of traumatic events.

Despite of high rates of adverse childhood experiences (ACEs) in pathological gamblers, researchers have rarely studied which types of ACEs often co-occur and how these profiles of ACEs are related to current psychopathology. We aimed to identify profiles of ACEs in pathological gamblers and examined how these profiles were related to gambling-related characteristics and current general psychopathology. In 329 current or lifetime pathological gamblers, diagnosed with the Composite Diagnostic Interview for DSM-IV, 10 types of ACEs were measured using the Adverse Childhood Experiences Questionnaire. Global psychopathology was assessed using the Symptom Checklist SCL-27. ACE profiles were identified using latent class analysis. Differences between ACE profiles in gambling-related characteristics and global psychopathology were analyzed using MANOVA. We found that four out of five gamblers (n=257, 78.1%) reported at least one ACE. Four distinct ACE profiles were identified: 'Low ACE', 'High ACE', 'Physical and emotional abuse', and 'Neglect'. The number of the fulfilled pathological gambling criteria and the severity of current global psychopathology differed between the ACE profiles: Gamblers with a 'High ACE' profile fulfilled more pathological gambling criteria and showed a more severe current psychopathology than gamblers of the 'Low ACE' profile. Gamblers with a 'Physical and emotional abuse' or an 'Emotion neglect' profile showed an intermediate severity of psychopathology. Our findings indicate that four different ACE profiles can be distinguished in pathological gamblers that differed in their gambling-related characteristics and current psychopathology. Systematic assessment of profiles of ACEs in pathological gamblers may inform about the severity of current global psychopathology that might be important to be addressed in addition to gambling-specific treatment., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Context: Exposure to traumatic events should be systematically assessed in health care services so that trauma-related treatment can be offered when appropriate. However, professionals often lack expertise in trauma inquiry and response, and therefore require training in this field., Objective: We aimed to determine whether the "Learning how to ask" training for trauma inquiry and response (Read, Hammersley, & Rudegeair, 2007) is effective in increasing health care professionals' trauma inquiry behavior., Method: 148 professionals working in outpatient substance use disorder (SUD) services were cluster-randomized into an intervention or a waiting control group. The intervention group received a 1-day training and a refresher session 3 months later, while the control group received no training. The primary outcome was the change from baseline in the frequency of asking clients about traumatic events. Secondary outcomes were professionals' evaluation of the training, knowledge, attitudes toward and confidence in trauma inquiry and response., Results: Change from baseline in the frequency of asking clients about traumatic events was significantly greater at 3-month and 6-month follow-up in the intervention group compared with the control group (b = 0.43, 95% CI [0.27, 0.59], p < .001). The training was positively evaluated by the participants. Knowledge, positive attitudes toward and confidence in trauma inquiry and response all showed significantly greater increases in the intervention group than in the control group., Conclusions: These findings suggest that health care professionals can acquire skills in trauma inquiry and response from short trainings, which may enhance systematic assessment of traumatic events. (PsycINFO Database Record, ((c) 2018 APA, all rights reserved).)

Background: The risk of developing a problem gambling behavior is distributed unequally among the population. For example, individuals who report stressful life events, show impairments of mental health or belong to a socio-economically deprived group are affected more frequently by gambling problems. The aim of our study is to investigate whether these risk factors are equally relevant for all gambling groups (social = 0 DSM-5 criteria, at risk = 1 DSM-5 criterion, problem = 2-3 DSM-5 criteria, disordered = 4-9 DSM-5 criteria). Methods: Of a total of 10,000 participants in the representative gambling survey in Austria in 2015, 4,082 individuals reported gambling during the last 12 months and were allocated to the four gambling groups according to DSM-5. With social gamblers as the reference group, relevant risk factors for the other three groups were identified by means of bi- and multivariate multinomial logistic regression. Results: Significant risk factors for gambling disorder are at-risk alcohol use (OR = 4.9), poor mental health (OR = 5.9), young age (≤26 years, OR = 2.1), a low level of formal education (OR = 2.4), having grown up with a single parent (OR = 2.5), parents with addiction problems (OR = 2.3) and belonging to the working class (OR = 2.9). Risk factors for problem gambling are parents with addiction problems (OR = 3.8), poor mental health (OR = 2.6) and a young age (OR = 2.2). With regard to at-risk gambling, only growing up with a single parent was relevant (OR = 2.4). Conclusion: Overall, the results of this study suggest, that the number and the influence of the included risk factors differ between gambling problem groups. Apparently, the development of severe gambling problems is to a lesser extent facilitated by specific risk factors than by their cumulative presence. Therefore, future prevention and treatment measures should place a particular focus on individuals who have experienced growing up in a difficult family situation, have poor mental health, suffer from substance-related problems or have a low level of formal education.

Background: In Germany, about 1.4 to 2.6 million people are dependent on prescription drugs, mainly benzodiazepine derivatives and opioid analgesics. Despite this large number of affected people, studies on prescription behaviour and drug intake are scarce. There are particularly few empirical findings available with regard to elderly people age 65 and over who are disproportionally affected by prescription drug abuse., Methods: The North German Pharmacy Computing Centre (Norddeutsches Apothekenrechenzentrum, NARZ) collects the prescription data of about 11 million citizens and covers over 80% of drug pharmacies in North Germany. Based on the data from NARZ, we evaluate person-related prescriptions of benzodiazepines, Z‑drugs and opioid analgesics. By means of incremental calculations, we determine the prevalence of prescription drug use, the long-term intake, the average daily dose of these medications and present this information as a five-year trend (2006-2010)., Results: In 2006, 10.6% of the members of public healthcare system were prescribed at least one of the medications under study. This proportion hardly changed within the five-year span. The share of patients with benzodiazepine prescription steadily decreased from 2006 (5.5%) to 2010 (5.1%), especially among elderly people. While the prevalence of Z‑drug prescriptions remained the same at about 1%, there was a slight increase in prescriptions of opioid analgesics with 5.4% in 2006 to 5.7% in 2010. The proportion of patients with long-term prescriptions decreased with regard to benzodiazepines (from 17.0 to 12.8%) and Z‑drugs (from 24.3 to 21.2%), but increased for opioid analgesics (from 19.2 to 21.2%)., Conclusion: The analytical method used in this study is an innovative epidemiological approach to evaluate person-related register data over the course of several years. Establishing a monitoring system of prescription drugs with potential for dependence may allow for a quicker identification of trends and initiation of appropriate measures.

To monitor progress toward maternal and neonatal tetanus elimination (MNTE) in Cambodia, we conducted a nationwide serosurvey of tetanus immunity in 2012. Multistage cluster sampling was used to select 2,154 women aged 15 to 39 years. Tetanus toxoid antibodies in serum samples were measured by gold-standard double-antigen enzyme-linked immunosorbent assay (DAE) and a novel multiplex bead assay (MBA). Antibody concentrations of ≥0.01 IU/ml by DAE or the equivalent for MBA were considered seroprotective. Estimated tetanus seroprotection was 88% (95% confidence interval [CI], 86 to 89%); 64% (95% CI, 61 to 67%) of women had antibody levels of ≥1.0 IU/ml. Seroprotection was significantly lower (P < 0.001) among women aged 15 to 19 years (63%) and 20 to 24 years (87%) than among those aged ≥25 years (96%), among nulliparous women than among parous women (71 versus 97%), and among those living in the western region than among those living in other regions (82 versus 89%). The MBA showed high sensitivity (99% [95% CI, 98 to 99%]) and specificity (92% [95% CI, 88 to 95%]) compared with DAE. Findings were compatible with MNTE in Cambodia (≥80% protection). Tetanus immunity gaps should be addressed through strengthened routine immunization and targeted vaccination campaigns. Incorporating tetanus testing in national serosurveys using MBAs, which can measure immunity to multiple pathogens simultaneously, may be beneficial for monitoring MNTE., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Collection of surveillance data is essential for monitoring and evaluation of public health programs. Integrated collection of household-based health data, now routinely carried out in many countries through demographic health surveys and multiple indicator surveys, provides critical measures of progress in health delivery. In contrast, biomarker surveys typically focus on single or related measures of malaria infection, HIV status, vaccination coverage, or immunity status for vaccine-preventable diseases (VPD). Here we describe an integrated biomarker survey based on use of a multiplex bead assay (MBA) to simultaneously measure antibody responses to multiple parasitic diseases of public health importance as part of a VPD serological survey in Cambodia. A nationally-representative cluster-based survey was used to collect serum samples from women of child-bearing age. Samples were tested by MBA for immunoglobulin G antibodies recognizing recombinant antigens from Plasmodium falciparum and P. vivax, Wuchereria bancrofti, Toxoplasma gondii, Taenia solium, and Strongyloides stercoralis. Serologic IgG antibody results were useful both for generating national prevalence estimates for the parasitic diseases of interest and for confirming the highly focal distributions of some of these infections. Integrated surveys offer an opportunity to systematically assess the status of multiple public health programs and measure progress toward Millennium Development Goals.

We investigated the effects of ultrasound-facilitated fixation on oral mucosal morphology. Bovine dorsal tongue and porcine buccal (cheek) mucosa were sonicated for 0, 5, 10 or 15 min in a modified methacarn fixative, then incubated at 25° C for 4 h. Initial mass, volume and density of each specimen were measured before and after treatment and fixation. Morphometric analysis of the scanning electron micrographs was used to quantify changes in mucosal structure and microtexture. Statistical methods were used to describe the relation between sonication time, tissue density and relative change in tissue density. Our results indicate a linear correlation between sonication time and density of the dorsal tongue specimens. The treatment caused contraction of the tongue tissue and expansion of the buccal mucosa. Differences between initial and final tongue densities and the relative change in tissue density of the tongue vs. buccal mucosa were statistically significant. Differences observed between initial and final buccal mucosa densities were not statistically significant. Changes in buccal mucosa density correlated inversely with sonication time by contrast to the tongue density, which was correlated directly with this factor. Our study illustrates that while preservation of mucosal morphology and biopolymers can be achieved by ultrasound-facilitated fixation, its effects on tissue density are both time-dependent and specific to certain regions of the mouth.

The Leydig insulin-like gene (Ley I-L), a member of the insulin-related gene family, is specifically expressed in pre- and postnatal Leydig cells of the testis and in postnatal theca cells of the ovary. To determine the functional region of the mouse Ley I-L promoter and factors controlling the Ley I-L gene expression, we used 2.1 kb of the 5'-flanking region of the mouse Ley I-L gene to generate chimeric constructs with the chloramphenicol acetyltransferase gene (CAT). Transient transfections of MA10 Leydig cells, LTK- fibroblasts, and F9 embryonic cells by a series of 5'-deleted mouse Ley I-L promoter-CAT constructs revealed that the sequence between nucleotides -157 to +4 directs the transcription of the reporter gene in MA10 but not in LTK- and F9 cells, indicating that the determinants of Leydig cell-specific expression reside within this region. Deoxyribonuclease I (DNase I) footprint analysis revealed that the sequences designated SF-1/1, SF-1/2, and SF-1/3 within three DNase I-protected regions are homologous to the consensus binding site of the steroidogenic factor-1 (SF-1). Competition and antibody studies showed that the three SF-1-binding sites in the Ley I-L promoter have similar binding affinities for SF-1. Furthermore, transient transfections of MA10 cells with mutant reporter constructs, in which SF-1/1 or both SF-1/2 and SF-1/3 were deleted, demonstrated that all three SF-1-binding sites are required for SF-1-mediated stimulation of Ley I-L transcription. Cotransfection of an SF-1-containing expression vector together with a Ley I-L promoter-CAT construct into HeLa cells, which lack the endogenous SF-1 protein, resulted in CAT gene transcription, which indicated that SF-1 can transactivate the Ley I-L promoter. These data demonstrate an essential role of SF-1 in transcriptional activation of the Ley I-L promoter.