HPV vaccination coverage and factors among American Indians in Cherokee Nation.

PURPOSE: We estimated human papillomavirus (HPV) vaccine initiation coverage among American Indian adolescents and identified factors associated with HPV vaccination among parents of these adolescents. METHODS: We developed, tested, and disseminated a survey to a random sample of 2,000 parents of American Indian adolescents aged 9-17 years who had accessed Cherokee Nation Health Services from January 2019 to August 2020. We used log-binomial regression to estimate the unadjusted and adjusted weighted prevalence proportion ratios (PPR) and 95% confidence intervals (CI) for adolescent HPV vaccine initiation. RESULTS: HPV vaccine initiation coverage (≥ 1 dose) was 70.7% among adolescents aged 13-17 years. The prevalence of HPV vaccine initiation was higher among American Indian adolescents whose parents were aware of the HPV vaccine (adjusted weighted PPR 3.41; 95% CI 2.80, 4.15) and whose parents received a recommendation from their provider (adjusted weighted PPR 2.70; 95% CI 2.56, 2.84). The most common reasons reported by parents to vaccinate their children were to protect them against HPV-associated cancers (25.7%) and receiving a recommendation from a healthcare provider (25.0%). Parents cited vaccine safety concerns as the main reason for not getting their children vaccinated (33.2%). CONCLUSIONS: HPV vaccine initiation coverage among American Indian adolescents in Cherokee Nation was consistent with the national survey estimates. However, allaying parental concerns about vaccine safety and encouraging providers to recommend the HPV vaccine could improve coverage.

Sensitivity and specificity of the Amyotrophic Lateral Sclerosis Functional Rating Scale-Revised to detect dysarthria in individuals with amyotrophic lateral sclerosis.

INTRODUCTION/AIMS: Given the widespread use of the Amyotrophic Lateral Sclerosis Functional Rating Scale-Revised (ALSFRS-R) to measure disease progression in ALS and recent reports demonstrating its poor sensitivity, we aimed to determine the sensitivity and specificity of the ALSFRS-R bulbar subscale and speech item to detect validated clinical ratings of dysarthria in individuals with ALS. METHODS: Paired ALSFRS-R and validated Speech Intelligibility Test (SIT) data from individuals with ALS were analyzed. Trained raters completed duplicate, independent, and blinded ratings of audio recordings to obtain speech intelligibility (%) and speaking rate (words per minute, WPM). Binary dysarthria profiles were derived (dysarthria ≤96% intelligible and/or <150 WPM). Data were obtained using the Kruskal-Wallis test, receiver-operating characteristic (ROC) curve, area under the curve (AUC), sensitivity and specificity percentages, and positive/negative predictive values (PPV/NPV). RESULTS: A total of 250 paired SIT and ALSFRS-R data points were analyzed. Dysarthria was confirmed in 72.4% (n = 181). Dysarthric speakers demonstrated lower ALSFRS-R bulbar subscale (8.9 vs. 11.2) and speech item (2.7 vs. 3.7) scores (P < .0001). The ALSFRS-R bulbar subscale score had an AUC of 0.81 (95% confidence interval [CI] 0.75 to 0.86). A subscale score of ≤11 yielded a sensitivity of 86%, specificity of 57%, PPV of 84%, and NPV of 60% to correctly identify dysarthria status. The ALSFRS-R speech item score demonstrated an AUC of 0.81 to detect dysarthria (95% CI 0.76 to 0.85), with sensitivity of 79%, specificity of 75%, PPV of 89%, and NPV of 58% for a speech item cutpoint of ≤3. DISCUSSION: The ALSFRS-R bulbar and speech item subscale scores may be useful, inexpensive, and quick tools for monitoring dysarthria status in ALS.

SPR perspectives: Environmental influences on Child Health Outcomes (ECHO) Program: overcoming challenges to generate engaged, multidisciplinary science.

The US National Institutes of Health-funded Environmental influences on Child Health Outcomes (ECHO) Program brings together 69 cohorts and over 57,000 children from across the nation to address five key pediatric outcome areas with high public health impact: pre-, peri-, and postnatal outcomes; upper and lower airway health; obesity; neurodevelopment; and positive health. We describe (1) the ECHO Program infrastructure that was designed to facilitate collaboration across over 1200 investigators and support the development of a cohort-wide data collection protocol and (2) the many challenges that were overcome in rapidly launching this large-scale program. Guided by a commitment to transparency, team science, and end user stakeholder engagement, ECHO successfully launched a unified study protocol and is working across disciplines to generate high-impact, solution-oriented research to improve children's lives for generations to come. IMPACT: Many children in the United States experience chronic health conditions or do not reach their developmental potential. The Environmental influences on Child Health Outcomes (ECHO) Program brings together 69 existing cohort studies comprising over 57,000 children to identify modifiable aspects of the early environment associated with pediatric outcomes with high public health impact: pre-, peri-, and postnatal outcomes; upper and lower airway health; obesity; neurodevelopment; and positive health. We describe the collaborative, team science-informed approach by which over 1200 investigators convened to form the ECHO Program and foster solution-oriented research to improve the health of children for generations to come.

Swallowing Safety and Efficiency Impairment Profiles in Individuals with Amyotrophic Lateral Sclerosis.

Dysphagia is common in individuals with amyotrophic lateral sclerosis (ALS) and associated with reductions in quality of life and health-related outcomes. Despite the high prevalence of dysphagia in ALS, functional impairment profiles of swallowing safety and efficiency have not been comprehensively examined. We therefore aimed to determine the relative prevalence of unsafe and inefficient swallowing in a large cohort of individuals with ALS. We further sought to examine the impact of global and bulbar disease progression (Amyotrophic Lateral Sclerosis Functional Rating Scale-Revised "ALSFRS-R" total and bulbar scores, respectively), disease duration, and onset type on swallowing impairment profiles. One hundred individuals with ALS completed a standardized videofluoroscopic swallowing examination Two independent and blinded raters performed validated ratings of safety (Penetration-Aspiration Scale, PAS) and efficiency (Analysis of Swallowing Physiology: Events, Kinematics, and Timing, ASPEKT % residue). Binary classifications of safety (unsafe: PAS ≥ 3), efficiency (inefficient: total residue ≥ 3% of C2-42) and global swallowing status were derived. The ALSFRS-R was completed to derive ALSFRS-R total and bulbar subscale scores. Demographic data (disease duration and onset type) for each participant was also recorded.  Descriptives, 2 × 2 contingency tables with Fishers exact test, and independent samples t-tests were performed (α = 0.05). Prevalence of unsafe and inefficient swallowing was 48% and 73%, respectively. Global swallowing profiles were, in rank order: unsafe and inefficient (39%), inefficient but safe (34%), safe and efficient (18%), and unsafe but efficient (9%). There were no differences in global disease progression or disease duration across swallowing impairment profiles. ALSFRS-R bulbar subscale scores were significantly lower in unsafe versus safe swallowers, p < 0.05. Spinal onset patients had a greater proportion of safe swallowers as compared to bulbar onset patients (p = 0.000, Fisher's exact test). Both spinal and bulbar onset patients demonstrated a higher prevalence of inefficient swallowers as compared to efficient swallowers (p = 0.04, Fisher's exact test). Dysphagia was prevalent in this group of individuals with ALS. Approximately half demonstrated safety impairments and two-thirds had impairments in swallowing efficiency. Inefficient swallowing was approximately four times more likely to be the initial functional impairment in patients with one pathophysiologic functional impairment. A longitudinal study is needed to examine the temporal evolution of dysphagia in ALS.

Discriminant Ability of the 3-Ounce Water Swallow Test to Detect Aspiration in Acute Postoperative Cardiac Surgical Patients.

Dysphagia is a common complication of cardiac surgery (CS) contributing to morbidity and mortality. Although early dysphagia detection is important, no current screening guidelines or validated tools exist in the cardiac intensive care setting. We therefore aimed to examine the discriminant ability of the 3-ounce water swallow test (3 oz. WST) to detect aspiration in acute postoperative CS patients. 196 postoperative CS patients were enrolled in this prospective single-center study. Participants completed the 3 oz. WST and a standardized Flexible Endoscopic Evaluation of Swallowing. Independent duplicate ratings of the penetration aspiration scale (PAS) were performed in a blinded fashion (100% agreement criteria). Receiver operating characteristic curve and area under the curve (AUC) analyses were performed with sensitivity, specificity, positive, and negative predictive values (PPV, NPV) derived. Fifty-four CS patients (28%) were confirmed aspirators (PAS ≥ 6), of whom 48% (n = 26) were silent aspirators (PAS = 8). Both the sensitivity and specificity of the 3 oz. WST to identify instrumentally confirmed aspiration was 63% (AUC: 0.63, 95% CI: 0.54, 0.72), and PPV was 39% and NPV 82%. The 3 oz. WST demonstrated fair discriminant ability to detect aspiration in acute postoperative CS patients. The high rate of silent aspiration may explain, in part, these findings given that the screening fail criteria include an overt cough response. In isolation, the 3 oz. WST does not represent a sensitive screen of aspiration in postoperative CS patients with a need to identify alternative screening tools for this setting.

Linking the Cherokee Nation Cancer Registry With Electronic Medical Records: A Tribal-Academic Center Partnership.

The Cherokee Nation Cancer Registry (CNCR) is the only tribally operated Surveillance, Epidemiology, and End Results program registry. As registries, including the CNCR, lack detailed data characterizing health behavior or comorbidity, we aimed to enrich the CNCR by linking it with Cherokee Nation's electronic medical record (EMR). We describe the process of a tribal-academic partnership and linking records between the CNCR and the EMR for American Indian people diagnosed with cancer from 2015 to 2020. Prior to data linkage, our team worked with the Cherokee Nation Governance Board and Institutional Review Board to ensure tribal data sovereignty was maintained. While not all persons in the CNCR receive health care at Cherokee Nation, 63% linked with an EMR. We observed differences (P < .001) between cancer site, year at diagnosis, age at diagnosis, and gender by EMR linkage status. Once we further validate linkages and assess data completeness, we will evaluate relationships between behavioral risk factors, comorbidities, and cancer outcomes.

Inactivation of virginiamycin by Aureobasidium pullulans.

OBJECTIVE: To test the inactivation of the antibiotic, virginiamycin, by laccase-induced culture supernatants of Aureobasidium pullulans. RESULTS: Fourteen strains of A. pullulans from phylogenetic clade 7 were tested for laccase production. Three laccase-producing strains from this group and three previously identified strains from clade 5 were compared for inactivation of virginiamycin. Laccase-induced culture supernatants from clade 7 strains were more effective at inactivation of virginiamycin, particularly at 50 °C. Clade 7 strain NRRL Y-2567 inactivated 6 µg virginiamycin/ml within 24 h. HPLC analyses indicated that virginiamycin was degraded by A. pullulans. CONCLUSIONS: A. pullulans has the potential for the bioremediation of virginiamycin-contaminated materials, such as distiller's dry grains with solubles (DDGS) animal feed produced from corn-based fuel ethanol production.

Management Team Perceptions of Risks and Strategies for Preventing Falls Among Short-Stay Patients in Nursing Homes.

We sought to understand strategies reported by members of the nursing home management team used to prevent falls in short-stay nursing home patients. Using Donabedian's model of structure, process, and outcomes, we interviewed 16 managers from 4 nursing homes in central North Carolina. Nursing home managers identified specific barriers to fall prevention among short-stay patients including rapid changes in functional and cognitive status, staff unfamiliarity with short-stay patient needs and patterns, and policies impacting care. Few interventions for reducing falls among short-stay patients were used at the structure level (eg, specialized units, workload ratio, and staffing consistency); however, many process-level interventions were used (eg, patient education on problem solving, self-care/mobility, and safety). We described several barriers to fall prevention among short-stay patients in nursing homes. From these descriptions, we propose three interventions that might reduce falls for short-stay patients and could be tested in future research: (1) clustering short-stay patients within a physical location to permit higher staff-patient ratios and enhanced surveillance, (2) population-based prevention interventions to supplement existing individually tailored prevention strategies (eg, toileting schedules, medication review for all), and (3) transitional care interventions that transmit key information from hospitals to nursing homes.

A population based caregivers profile and training needs assessment in Oklahoma.

Background and Objectives: Studies indicate an expected population growth of almost fifty percent in Oklahomans aged 65 and older by 2030. According to the United Health Foundation, Oklahoma ranked 48th in overall senior health in 2017. Research Design and Methods: The Oklahoma Healthy Aging Initiative administered a Consumer Needs Assessment Survey by mail to a stratified random sample of the 475,518 registered voters aged 65 and older. The survey was anonymous and stratified by region. The survey contained six sections: introduction, health and health promotion, activities/recreation, information and assistance, caregiving and "about you." Results: Nearly one in three (32%) of respondents indicated that they directly or indirectly provide care to another, with another 9% responding they maybe provide care, and the remaining 59% responding no. Nearly 10% of people who say they are not caregivers reported that they participate at least one day a week in caring for a sick or invalid spouse, family member, or friend living with them, indicating current estimates of the number of caregivers is low. Discussion and Implications: Those who report they are or are maybe caregivers tend to be more interested in community events and more interested in caregiver respite. In addition, maybe caregivers appear to be more interested in health improvement topics and classes, such as health and wellness, mental health, chronic disease, and computers when compared to both caregivers and non-caregivers. Our survey results indicate a need for caregivers to receive respite services as well as training courses in Oklahoma communities.

Novel Feruloyl Esterase from Lactobacillus fermentum NRRL B-1932 and Analysis of the Recombinant Enzyme Produced in Escherichia coli.

UNLABELLED: A total of 33 Lactobacillus strains were screened for feruloyl esterase (FE) activity using agar plates containing ethyl ferulate as the sole carbon source, and Lactobacillus fermentum NRRL B-1932 demonstrated the strongest FE activity among a dozen species showing a clearing zone on the opaque plate containing ethyl ferulate. FE activities were monitored using high-performance liquid chromatography with an acetonitrile-trifluoroacetic acid gradient. To produce sufficient purified FE from L. fermentum strain NRRL B-1932 (LfFE), the cDNA encoding LfFE (Lffae) was amplified and cloned by using available closely related genome sequences and overexpressed in Escherichia coli A 29.6-kDa LfFE protein was detected from the protein extract of E. coli BL21(pLysS) carrying pET28bLffae upon IPTG (isopropyl-ß-d-thiogalactopyranoside) induction. The recombinant LfFE containing a polyhistidine tag was purified by nickel-nitrilotriacetic acid affinity resin. The purified LfFE showed strong activities against several artificial substrates, including p-nitrophenyl acetate and 4-methylumbelliferyl p-trimethylammoniocinnamate chloride. The optimum pH and temperature of the recombinant LfFE were around 6.5 and 37°C, respectively, as determined using either crude or purified recombinant LfFE. This study will be essential for the production of the LfFE in E. coli on a larger scale that could not be readily achieved by L. fermentum fermentation. IMPORTANCE: The production of feruloyl esterase (FE) from Lactobacillus fermentum NRRL B-1932 reported in this study will have immense potential commercial applications not only in biofuel production but also in pharmaceutical, polymer, oleo chemical, cosmetic additive, and detergent industries, as well as human health-related applications, including food flavoring, functional foods, probiotic agents, preventive medicine, and animal feed. Given the essential role FE plays in the production of hydroxycinnamic acids and ferulic acid, plus the generally regarded as safe status of lactobacilli, which therefore have less regulatory concerns, LfFE from the probiotic L. fermentum reported in this work can be directly used for increased production of high-value hydroxycinnamates and ferulic acid from natural or synthetic carbon sources.

Phylogenetic classification of Aureobasidium pullulans strains for production of feruloyl esterase.

OBJECTIVE: The objective was to phylogenetically classify diverse strains of Aureobasidium pullulans and determine their production of feruloyl esterase. RESULTS: Seventeen strains from the A. pullulans literature were phylogenetically classified. Phenotypic traits of color variation and endo-ß-1,4-xylanase overproduction were associated with phylogenetic clade 10 and particularly clade 8. Literature strains used for pullulan production all belonged to clade 7. These strains and 36 previously classified strains were tested for feruloyl esterase production, which was found to be associated with phylogenetic clades 4, 11, and particularly clade 8. Clade 8 strains NRRL 58552 and NRRL 62041 produced the highest levels of feruloyl esterase among strains tested. CONCLUSIONS: Production of both xylanase and feruloyl esterase are associated with A. pullulans strains in phylogenetic clade 8, which is thus a promising source of enzymes with potential biotechnological applications.

Racial, Ethnic, and Age Differences in the Incidence and Survival of Childhood Cancer in Oklahoma, 1997-2012.

While cancer is relatively rare in children under 20, it is the leading cause of disease-related death among children aged 5 to 14 years. We aimed to describe the incidence and survival of childhood cancer in Oklahoma from 1997-2012. We calculated age-adjusted incidence rates and five-year observed survival by cancer type using Oklahoma Central Cancer Registry and Surveillance, Epidemiology, and End Results program data among children diagnosed with cancer under the age of 20 from 1997-2012. The average annual age-adjusted incidence rate of childhood cancer was 168.9 per million for the US and 171.7 per million for Oklahoma. Overall, Oklahoma had lower survival from childhood cancer compared to the US (77.0% v. 80.6%). In recent years, research has been conducted on the epidemiology of childhood cancer. Little research has been done, however, on the incidence or survival of childhood cancer at state levels and none focused exclusively on Oklahoma.

Five-Year Cancer Survival Rates in Oklahoma from 1997 to 2008.

INTRODUCTION: This study evaluated the five-year observed survival rates of American Indians/Alaskan Native, African American, and white cancer patients among various demographic characteristics in Oklahoma focusing on lung and bronchus, colon and rectum, female breast, and prostate for the cancer patients diagnosed between 1997 and 2008. METHODS: The five-year observed survival rates were calculated for overall cancer and specific cancer sites, using Kaplan-Meier method with data from the Oklahoma Central Cancer Registry. RESULTS: Overall, 51.5% patients diagnosed with cancer survived for five years. For specific sites we found: 79.2% for female breast cancer survived; 77.5% for prostate cancer; 12.9% for lung and bronchus cancer; and 49.9% for colorectal cancer. CONCLUSIONS: The five-year observed survival rates in Oklahoma were consistent with national trends. Overall, cancer survival seems to be improving over time, but there remains disparity with the AA and AI/AN populations in contrast to whites in Oklahoma.

Circulating Tumor Cells After Neoadjuvant Chemotherapy in Stage I-III Triple-Negative Breast Cancer.

BACKGROUND: Triple-negative breast cancer (TNBC) is characterized by a lack of estrogen and progesterone receptor expression and HER-2 gene amplification. Circulating tumor cells (CTCs) can be identified in 25 % of nonmetastatic breast cancer patients, and the identification of ≥1 CTC predicts outcome. This study was designed to determine whether CTCs present after neoadjuvant chemotherapy (NACT) predicted worse outcome in nonmetastatic TNBC patients. METHODS: CTCs were assessed in 57 TNBC patients with nonmetastatic TNBC after the completion of NACT. CTCs (per 7.5 ml blood) were identified using the Cell Search(®) System (Janssen). Log-rank test and Cox regression analysis were applied to establish the association of CTCs with relapse-free (RFS) and overall survival (OS). RESULTS: Median follow-up was 30 months, and mean age was 53 years. Fifty-four patients (95 %) had >2-cm tumors, 42 (84 %) were nuclear grade 3, and 42 (74 %) had positive axillary lymph nodes. One or more CTC was identified in 30 % of patients. CTC presence was not associated with primary tumor size, high grade, or lymph node positivity. Multivariate analysis demonstrated that detection of ≥1 CTC predicted decreased RFS (log-rank P = 0.03, HR 5.25, 95 % CI 1.34-20.56) and OS (log-rank P = 0.03, HR 7.04, 95 % CI 1.26-39.35). CONCLUSIONS: One or more CTCs present after NACT predicted relapse and survival in nonmetastatic TNBC patients. This information would be helpful in future clinical trial design of adjuvant treatments for TNBC patients who are at risk for relapse after completing NACT.

Antibacterial activity of a cell wall hydrolase from Lactobacillus paracasei NRRL B-50314 produced by recombinant Bacillus megaterium.

The cell-free supernatant (CFS) from Lactobacillus paracasei NRRL B-50314 culture has been previously reported as containing antibacterial activity against a wide variety of Gram-positive bacteria. The CFS protein gel slice corresponding to antibacterial activities was subjected to trypsin digestion and ion trap MASS (Gel/LC-MS/MS) analysis. BlastP search of the resulted IQAVISIAEQQIGKP sequence led to a hypothetical cell-wall associated hydrolase (designated as CWH here) from Lactobacillus paracasei ATCC 25302. Further analyses of CWH revealed that the IQAVISIAEQQIGKP belongs to a highly conserved region of the NlpC/P60 superfamily. The L. paracasei NRRL B-50314 CWH gene, cloned in pStrepHIS1525CWH477, was introduced into Bacillus megaterium MS 941. The production of CWH477 protein was induced by xylose. The CWH477 protein was purified by using NiNTA column, and elution fraction E2 showed highest antibacterial activity. This study and bioinformatics analyses suggested that the antibacterial activity of CWH could originate from its cell wall degrading enzymatic function.

Circulating tumor cells in non-metastatic triple-negative breast cancer.

Circulating tumor cells (CTCs) can be identified in approximately 25 % of stage I-III breast cancer patients; CTCs presence is a predictor of poor outcome in metastatic breast cancer, but little is known regarding the prognostic significance of CTCs in non-metastatic triple-negative breast cancer (TNBC) patients. The aim of this study was to determine whether CTCs predict worse outcome in non-metastatic TNBC patients. We evaluated CTCs in 113 patients with stages I-III TNBC at the time of definitive surgery. CTCs were assessed using the CellSearch System®. Progression-free and overall survival were defined as time elapsed between date of diagnosis and either date of clinical disease progression, death, or last follow-up. Log-rank test and Cox regression analysis were used to determine associations of CTCs with progression-free and overall survival. The median follow-up was 40 months. CTCs were identified in 23/113 (20 %) of patients. No primary tumor characteristic or lymph node status predicted the presence of CTCs. The identification of ≥2 CTCs predicted shorter progression-free (log rank P ≤ 0.001; hazard ratio 8.30, 95 % CI 2.61-26.37) and overall survival (log rank P = 0.0004; hazard ratio 7.19, 95 % CI 1.98-26.06) versus survival for patients with <2 CTCs. Two or more CTCs predict shorter progression-free and overall survival in TNBC patients. Larger studies are needed to determine whether CTC assessment provides beneficial information that could be used in stratifying TNBC patients at increased risk for disease progression. Finally, CTCs characterization could facilitate the development of novel treatment approaches for TNBC.

Strategies for Managing Difficult Clinical Situations in Between Sessions.

Therapists often have to deal with situations that make them uneasy to leave their clients on their own. They can go home and wonder what their clients are doing, whether they are safe, have a place to stay, even whether or not they are fighting. These thoughts can be overwhelming for the therapist. They may think about their clients' safety, they may also be worried about their own professional way of dealing with these situations, and its implications for them as clinician. The aim of this paper is to assist therapists to manage their own anxiety and to help them help their clients in between sessions in difficult situations such as suicidality, child abuse and neglect, intimate partner violence, crisis and stress management.

Perception of Barriers to and Factors Associated with HPV Vaccination Among Parents of American Indian Adolescents in the Cherokee Nation.

The purpose of this study was to understand the perceptions of HPV vaccination barriers and factors among parents or guardians of American Indian adolescents in the Cherokee Nation. Fifty-four parents of American Indian adolescents in the Cherokee Nation participated in one of eleven focus group discussions from June to August 2019. Discussions were recorded, transcribed, coded, and analyzed for themes. Protection against cancer was the primary parent-reported reason for vaccinating their children against HPV. The lack of information and safety concerns about the HPV vaccine were the main reasons for non-vaccination. To increase HPV vaccine uptake, parents strongly supported offering vaccinations in school. Furthermore, increased healthcare provider-initiated discussion can ease parental concerns about HPV vaccine safety and improve coverage.

Combinatorial evaluation of laccase-mediator system in the oxidation of veratryl alcohol.

Laccases play an important role in the biological break down of lignin and have great potential in the deconstruction of lignocellulosic feedstocks. We examined 16 laccases, both commercially prepared and crude extracts, for their ability to oxidize veratryl alcohol in the presence of various solvents and mediators. Screening revealed complete conversion of veratryl alcohol to veratraldehyde catalyzed by a crude preparation of the laccase from Trametes versicolor ATCC 11235 and the mediator TEMPO in 20 % (v/v) tert-butanol.

Lipase production by diverse phylogenetic clades of Aureobasidium pullulans.

Thirty-nine strains representing 12 diverse phylogenetic clades of Aureobasidium pullulans were surveyed for lipase production using a quantitative assay. Strains in clades 4 and 10 produced 0.2-0.3 U lipase/ml, while color variant strain NRRL Y-2311-1 in clade 8 produced 0.54 U lipase/ml. Strains in clade 9, which exhibit a dark olivaceous pigment, produced the highest levels of lipase, with strain NRRL 62034 yielding 0.57 U lipase/ml. By comparison, Candida cylindracea strain NRRL Y-17506 produced 0.05 U lipase/ml under identical conditions. A. pullulans strain NRRL 62034 reached maximal lipase levels in 5 days on lipase induction medium, while A. pullulans strain NRRL Y-2311-1 and strains in clades 4 and 10 were highest after 6 days. A. pullulans strain NRRL Y-2311-1 and strains in clade 9 produced two extracellular proteins in common, at >50 and <37 kDa.