Utility of Dried Blood Spots for the Diagnosis of Congenital Cytomegaloviruses within the First 21 Days of Life in a Single Center.

In this retrospective study, we aimed to evaluate the performance of dried-blood-spot (DBS) testing as a diagnostic method for the congenital cytomegalovirus (cCMV). We reviewed the medical records and DBS test results of 89 patients who had also undergone diagnostic cCMV testing within the first 21 days of life. The DBS test had a sensitivity of 83.9%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 73%. Patients with a true-positive DBS had a higher median level of CMV in blood according to PCR than those with a false-negative result. Additionally, all patients with cCMV and hearing loss had a positive DBS test, with higher median viremia levels observed in those with hearing loss compared to those without a CMV PCR blood test. These results suggest that DBS-based testing is useful in the diagnosis of cCMV, and its performance may be related to levels of CMV viremia. DBS testing accurately identified those patients with congenital/early onset hearing loss and those at risk of developing late-onset hearing loss.

The Impact of Asymptomatic Congenital Cytomegalovirus on Adult Quality of Life.

OBJECTIVE: Children born with asymptomatic congenital cytomegalovirus infection (AcCMV) have increased risk for hearing loss, which may affect their quality of life into adulthood. We aim to determine quality of life outcomes among adults who were identified at birth with AcCMV compared with controls, using the cohort of the Houston Congenital CMV Longitudinal Study. METHODS: Quality of life was determined using the self-reported Quality of Life Inventory (QOLI). Sixty-one of 109 AcCMV subjects and 23 of 51 controls completed QOLI. Percentile scores of subjects were compared with percentile scores of controls using Student t tests. QOLI percentile scores were compared among AcCMV subjects with (N = 14) and without hearing loss (N = 47). RESULTS: There was no difference in mean percentile scores on QOLI between AcCMV subjects (59.8 [SD = 27.6]) and controls (57.3 [SD = 35.3]; p = 0.754). Percentile scores indicate an average overall quality of life classification for AcCMV subjects and controls. There was no difference in mean percentile scores on the QOLI between AcCMV subjects with and without hearing loss (54.8 [SD = 25.2]) and 61.3 [SD = 28.3]; p = 0.440, respectively). CONCLUSION: Adults born with AcCMV do not seem to have lower ratings of quality of life compared with uninfected controls. Although our study had small sample size, hearing loss does not seem to be a significant predictor of QOLI percentile scores among AcCMV subjects. Quality of life in adulthood does not seem to be affected by an individual's awareness of screening positive for CMV, which supports the notion of "no harm" occurring from universal newborn screening for congenital CMV infection.

Maternal cytomegalovirus immune status and hearing loss outcomes in congenital cytomegalovirus-infected offspring.

OBJECTIVES: The purpose of this study is to determine the relationship between maternal primary and recurrent CMV infection during pregnancy, symptoms at birth in the newborn, and long term hearing loss through18 years of age. PATIENTS AND METHODS: 237 mother-infant pairs in the Houston, Texas area identified through maternal CMV IgG and IgM antibody serologic screening and newborn screening using urine CMV culture to identify congenital CMV infection were enrolled in the Houston Congenital CMV Longitudinal Study. Mothers were categorized as having primary or recurrent or unknown maternal CMV infections, and newborns were categorized at birth as having symptomatic or asymptomatic congenital CMV infection, or as uninfected controls. All three newborn groups were followed longitudinally with serial hearing evaluations up to 18 years of age. The relationship between type of maternal CMV infection, newborn classification, and the occurrence of hearing loss over time was determined through Kaplan-Meier survival analysis, life table analysis, and a simulated ascertainment of maternal infection type for the unknown categories. RESULTS: Of 77 newborns with symptomatic congenital CMV infection, 12 (16%) of mothers had a primary CMV infection during pregnancy; 4 (5%) had a non-primary infection, and the type of infection in 48 (79%) could not be determined and were classified as unknown type of maternal infection. Fifty Seven (74%) of the 77 symptomatic children had hearing loss by 18 years of age, including 9 of the 12 (75%) who were born to mothers with primary infection and 48 (79%) of the 61 with unknown type of maternal infection. Of the 109 newborns with asymptomatic congenital CMV infection, 51 (47%) were born to mothers with a primary CMV infection during pregnancy, 18 (17%) to mothers with a recurrent infection; and 40 (37%) had unknown type of infection. Of these 109 asymptomatic cases, 22 (20%) developed hearing loss, including 14 out of 51 (28%) of those born to mothers with primary infection, two out of the 18 (11%) born to mothers with recurrent infection, and 6 out of the 40 (15%) to mothers of unknown infection type. Of the 51 uninfected newborn controls, 10 (20%) of mothers had a primary CMV infection during pregnancy, 5 (10%) had a non-primary infection, 10 (20%) were never infected, and 26 (51%) were assigned unknown type of infection. Three controls (6%) developed hearing loss, with 1 being born to a mother with primary infection and 1 to a mother never infected with CMV. CONCLUSIONS: Both primary and non-primary maternal CMV infections during pregnancy resulted in symptomatic and asymptomatic congenital CMV infection. Symptomatic congenital CMV infection was more likely to occur after primary maternal CMV infection. Sensorineural hearing loss occurred in children born to mothers with both primary and non-primary CMV infections, and in both asymptomatic and symptomatic congenital CMV infection, but was more common after maternal primary infection. Most, but not all, hearing loss in children with cCMV associated hearing loss was first detected within the first year of life.

Inattention and Hyperactivity in Children with Symptomatic and Asymptomatic Congenital Cytomegalovirus.

OBJECTIVES: To explore the relationship between congenital cytomegalovirus (CMV) and inattention and hyperactivity among school-aged children. METHODS: The Behavior Assessment System for Children, Second Edition, parent- and self-report, was completed among children with symptomatic congenital CMV (ScCMV) (n = 36), asymptomatic congenital CMV (AcCMV) (n = 76), and controls (n = 29) enrolled in a longitudinal cohort. The proportions of children with ScCMV, AcCMV, and controls with Attention Problems or Hyperactivity T-scores ever ≥ 65 were compared. Mean T-scores in these domains were also compared and adjusted for IQ. RESULTS: Children with AcCMV did not differ from controls in the proportion of children with elevated Attention Problems or Hyperactivity T-scores or in mean Attention Problems or Hyperactivity T-scores. Children with ScCMV had a higher proportion of elevated Attention Problems T-scores compared with the AcCMV group but not controls. There were no differences in the proportions of children with elevated Hyperactivity T-scores between ScCMV and AcCMV or control groups. Children with ScCMV had higher mean Attention Problems T-scores versus those with AcCMV and controls and higher mean Hyperactivity T-scores versus those with AcCMV but not controls. After adjustment for IQ, differences in mean Attention Problems or Hyperactivity T-scores were no longer significant. CONCLUSION: Children with AcCMV are not at increased risk of inattention or hyperactivity compared with controls. However, our study suggests an increased prevalence of inattention and hyperactivity among children with ScCMV. Differences in IQ were confirmed to have a confounding effect. Evaluation for attention-deficit/hyperactivity disorder may be warranted in this population.

Cortical Visual Impairment in Congenital Cytomegalovirus Infection.

PURPOSE: To describe the presentation, evolution, and long-term outcome of cortical visual impairment (CVI) in patients with symptomatic congenital cytomegalovirus (CMV) infection, and to identify risk factors for the development of CVI in patients with symptomatic congenital CMV. METHODS: Retrospective subanalysis of a long-term prospective cohort study with data gathered from 1982 to 2013. RESULTS: Eleven of 77 (14.3%) patients with symptomatic CMV, 0 of 109 with asymptomatic CMV, and 51 control patients had CVI. Overall, patients with symptomatic CMV had worse vision than patients with asymptomatic CMV, who in turn had worse vision than control patients. Microcephaly, intracranial calcification, dilatation of ventricles, encephalomalacia, seizure at birth, optic atrophy, chorioretinitis/retinal scars, strabismus, and neonatal onset of sensorineural hearing loss were risk factors associated with CVI. CONCLUSIONS: CVI may result from symptomatic congenital CMV infection. The relationship of CVI and its risk factors in patients with CMV suggests the potential to predict the development of CVI through predictive modeling in future research. Early screening of CVI in children born with symptomatic congenital CMV can facilitate educational, social, and developmental interventions. [J Pediatr Ophthalmol Strabismus. 2019;56(3):194-202.].

Intelligence and Academic Achievement With Asymptomatic Congenital Cytomegalovirus Infection.

OBJECTIVES: To examine intelligence, language, and academic achievement through 18 years of age among children with congenital cytomegalovirus infection identified through hospital-based newborn screening who were asymptomatic at birth compared with uninfected infants. METHODS: We used growth curve modeling to analyze trends in IQ (full-scale, verbal, and nonverbal intelligence), receptive and expressive vocabulary, and academic achievement in math and reading. Separate models were fit for each outcome, modeling the change in overall scores with increasing age for patients with normal hearing (n = 78) or with sensorineural hearing loss (SNHL) diagnosed by 2 years of age (n = 11) and controls (n = 40). RESULTS: Patients with SNHL had full-scale intelligence and receptive vocabulary scores that were 7.0 and 13.1 points lower, respectively, compared with controls, but no significant differences were noted in these scores among patients with normal hearing and controls. No significant differences were noted in scores for verbal and nonverbal intelligence, expressive vocabulary, and academic achievement in math and reading among patients with normal hearing or with SNHL and controls. CONCLUSIONS: Infants with asymptomatic congenital cytomegalovirus infection identified through newborn screening with normal hearing by age 2 years do not appear to have differences in IQ, vocabulary or academic achievement scores during childhood, or adolescence compared with uninfected children.

Genomic analysis of melanoma evolution following a 30-year disease-free interval.

Ultra-late melanoma recurrence is infrequent, poorly understood and, in most cases, difficult to unambiguously distinguish from a new primary melanoma. We identified a patient with a second melanoma diagnosed after a 30-year disease-free interval, and sought to determine if this new lesion was a recurrence of the original melanoma. Here we report the genomic sequence analysis of the exomes of 2 melanoma lesions isolated from the same individual in 1985 and 2015, and their comparison to each other and to the germline DNA of the patient. Identification of many shared somatic mutations between these lesions proves a lineal relationship spanning 30 years. Unlike prior reports of ultra-late melanoma recurrence, the availability of the original tumor and the use of comprehensive genomic analysis allowed us to confirm that the second lesion is truly a recurrence. We demonstrate the acquisition of numerous additional mutations during the 3 decade asymptomatic period. These data highlight the low but very long-lasting risk of recurrence in this patient population.

Long-term Visual and Ocular Sequelae in Patients With Congenital Cytomegalovirus Infection.

BACKGROUND: Cytomegalovirus (CMV) is the most common congenital viral infection in the United States. Visual and ocular sequelae in adolescents and adults who are congenitally infected with CMV have not been well studied. Better understanding of the long-term visual and ocular sequelae can help with early detection, intervention and appropriate educational accommodations. METHODS: This study evaluated 237 patients (77 symptomatic, 109 asymptomatic and 51 control) who underwent a series of age-appropriate ophthalmologic, audiologic and neurodevelopmental examinations from 1982 to 2013. The frequency and etiology of visual impairment and other nonophthalmologic findings were recorded for each patient. Ophthalmologic findings were tabulated, and risk factors for abnormalities were analyzed. RESULTS: Fourteen of the 77 (18.2%) symptomatic and none of the asymptomatic and control subjects had severe visual impairments (P ≤ 0.006). Moderate visual impairment did not differ between symptomatic and asymptomatic subjects. Three asymptomatic subjects had retinal scars. The most common visual or ocular sequelae in the symptomatic group were strabismus (23.4%), chorioretinal scars (19.5%), cortical visual impairment (14.3%), nystagmus (14.3%) and optic nerve atrophy (11.7%). Three symptomatic patients had delayed visual deterioration because of later occurring retinal disorders: peripheral retinal scar, rhegmatogenous retinal detachment and Coats' disease. CONCLUSION: Symptomatic CMV patients experienced more ophthalmologic sequelae and significantly worse visual outcomes than asymptomatic CMV and control patients. Later occurring retinal disorders were found in symptomatic patients, and there is no clear evidence that CMV can reactivate in the retinas of children who were congenitally infected. Major risk factors for severe visual impairment included symptomatic status, optic nerve atrophy, chorioretinitis, cortical visual impairment and sensorineural hearing loss.

Hearing Loss in Children With Asymptomatic Congenital Cytomegalovirus Infection.

OBJECTIVES: To assess the prevalence, characteristics, and risk of sensorineural hearing loss (SNHL) in children with congenital cytomegalovirus infection identified through hospital-based newborn screening who were asymptomatic at birth compared with uninfected children. METHODS: We included 92 case-patients and 51 controls assessed by using auditory brainstem response and behavioral audiometry. We used Kaplan-Meier survival analysis to estimate the prevalence of SNHL, defined as ≥25 dB hearing level at any frequency and Cox proportional hazards regression analyses to compare SNHL risk between groups. RESULTS: At age 18 years, SNHL prevalence was 25% (95% confidence interval [CI]: 17%-36%) among case-patients and 8% (95% CI: 3%-22%) in controls (hazard ratio [HR]: 4.0; 95% CI: 1.2-14.5; P = .02). Among children without SNHL by age 5 years, the risk of delayed-onset SNHL was not significantly greater for case-patients than for controls (HR: 1.6; 95% CI: 0.4-6.1; P = .5). Among case-patients, the risk of delayed-onset SNHL was significantly greater among those with unilateral congenital/early-onset hearing loss than those without (HR: 6.9; 95% CI: 2.5-19.1; P < .01). The prevalence of severe to profound bilateral SNHL among case-patients was 2% (95% CI: 1%-9%). CONCLUSIONS: Delayed-onset and progression of SNHL among children with asymptomatic congenital cytomegalovirus infection continued to occur throughout adolescence. However, the risk of developing SNHL after age 5 years among case-patients was not different than in uninfected children. Overall, 2% of case-patients developed SNHL that was severe enough for them to be candidates for cochlear implantation.

Self-assessed emergency readiness and training needs of nurses in rural Texas.

OBJECTIVE: Nurses, particularly public health nurses, play a key role in emergency preparedness and response in rural areas. To prepare rural jurisdictions for unforeseen disastrous events it is imperative to assess the public health emergency readiness and training needs of nurses. The objective of this study was to assess the self-reported terrorism preparedness and training needs of a nurse workforce. DESIGN AND SAMPLE: Cross-sectional prevalence of practicing nurses in regions of North Texas. 3,508 rural nurses practicing in North Texas participated in the study. MEASUREMENTS: Data were collected through a mailed survey; analyses included multinominal logistic regression and descriptive statistics. RESULTS: A total of 941 (27%) nurses completed the survey. The majority of respondents reported limited bioterrorism-related training. Fewer than 10% were confident in their ability to diagnose or treat bioterrorism-related conditions. Although only 30% expressed a willingness to collaborate with state and local authorities during a bioterrorism event, more than 69% indicated interest in future training opportunities. Preferred training modalities included small group workshops with instructor-led training, and Internet-based training. CONCLUSIONS: Licensing agencies, professional organizations, and community constituencies may need to play a stronger role in improving the bioterrorism-related emergency preparedness of rural nurses.

Resistance among cultivated sunflower germplasm to stem-infesting pests in the central Great Plains.

A 7-yr field study evaluated 61 oilseed sunflower, Helianthus annuus L., accessions and 31 interspecific crosses for resistance to attack by naturally occurring populations of three stem-infesting pests, the sunflower stem weevil, Cylindrocopturus adspersus (LeConte) (Coleoptera: Curculionidae); a longhorned beetle, Dectes texanus LeConte (Coleoptera: Cerambycidae); and a root boring moth, Pelochrista womonana (Kearfott) (Lepidoptera: Tortricidae), at two locations in the central Great Plains. Germplasm with potential sources of resistance to attack from all three stem-infesting species were revealed. Accessions PI 650558, PI 386230, and PI 431516 were consistent in averaging low densities of stem weevil larvae per stalk among lines tested, and PI 497939 exceeded 25 weevil larvae per stalk in only 1 yr of 5 yr of trials. Several interspecific crosses also had consistently low densities of C. adspersus larvae per stalk. Populations of both D. texanus and P. womonana were variable over years, but differences among the lines tested were evident in many trials, revealing potential for developing resistant germplasm. Four accessions (PI 386230, PI 431542, PI 650497, and PI 650558) had low larval densities of C. adspersus and P. womonana in addition to reduced percentage infestation by D. texanus. Results showed potential for developing resistant genotypes for these pests. The prospect of adding host plant resistance as an integrated pest management (IPM) tactic would provide another tool for reducing economic losses from stem-infesting insect pests of sunflower in the central Great Plains.

A clinical analysis of shoulder and hip joint infections in children.

BACKGROUND: Septic arthritis of the shoulder is a rare infection in healthy children. This infection requires prompt surgical drainage and antibiotic treatment. A delay in surgical intervention can result in damage to the articular surface of the glenohumeral joint, adjacent osteomyelitis, and possible growth disturbance. The clinical course of septic arthritis of the shoulder was compared with that of septic arthritis of the hip, a more common disease in children. METHODS: We identified 9 children with infections of the glenohumeral joint who presented to our pediatric hospital between 2001 and 2007. The average age at presentation was 7 years (range: 7 mo to 12 y). These patients were compared with 14 selected patients treated for septic arthritis of the hip (mean age 7 y, range: 1 to 12 y). Surgical drainage was performed by open arthrotomy in each case. A retrospective review and analysis of the medical records, laboratory tests, and radiographs of these patients were performed. RESULTS: Children with shoulder infections differed significantly (P<0.05) from patients with hip infections with regard to temperature, white blood cell count, and erythrocyte sedimentation rate at the time of admission. The average time from the onset of symptoms to presentation was notably longer in the shoulder group compared with the hip group (P=0.012). Adjacent osteomyelitis was found in 67% of the shoulders and 36% of the hips (P=0.214). Children suffering from septic arthritis of the shoulder showed higher rates of repeat surgical drainage (P=0.056) and extended hospitalizations (P=0.028). The total duration of antibiotics was longer in the shoulder group (P=0.059). CONCLUSIONS: Septic arthritis of the shoulder in the pediatric population often has a delayed presentation with a more complicated disease course than an infection of the hip. Children with shoulder infections require a longer duration of treatment and may experience a higher likelihood of skeletal complications. LEVEL OF EVIDENCE: Level III, retrospective comparative study.

Assessing bioterrorism preparedness and response of rural veterinarians: experiences and training needs.

Veterinarians play a unique role in emergency preparedness and response, and federal agencies and academic institutions therefore allocate considerable resources to provide training to enhance their readiness. However, the level of preparedness of veterinarians in many rural regions is yet to be improved. This article reports an assessment of the bioterrorism preparedness, specifically the experience and training needs, of rural veterinarians in North Texas. The study employed a cross-sectional design with a study population that included all veterinarians (N = 352) in the 37 counties within Texas Department of State Health Services Regions 2 and 3. Data on veterinarians practicing or residing in the target region were obtained from the Texas State Board of Veterinary Medical Examiners. The response rate was 35% (n = 121). Results indicate that chemical exposure was the condition most frequently seen and treated, followed by botulism and anthrax. The majority (80%) of respondents indicated that they had not previously participated in training related to bioterrorism preparedness, and many (41%) also indicated a willingness to participate in a state health department-initiated bioterrorism response plan. However, only 18% were confident in their ability to diagnose and treat bioterrorism cases. These results suggest that many North Texas veterinarians practicing in rural regions could benefit from additional training in bioterrorism preparedness and response. An area in particular need of further training is the diagnosis and treatment of Category A agents. Federal, state, and local health agencies are urged to increase training opportunities and to make additional efforts to involve veterinarians in bioterrorism preparedness and response.

Reducing liver cancer disparities: a community-based hepatitis-B prevention program for Asian-American communities.

OBJECTIVES: Several Asian-American groups are at a higher risk of dying of liver diseases attributable to hepatitis-B infection. This culturally diverse community should be well informed of and protected against liver diseases. The present study assesses the knowledge of hepatitis B before and after a hepatitis-B educational program and determines the infection status of an Asian community. METHODS: Nine Asian communities of Montgomery County, MD, enrolled in the hepatitis-B prevention program between 2005 and 2006. They attended culturally tailored lectures on prevention, completed self-administered pre- and posttests, and received blood screening for the disease. RESULTS: More than 800 Asian Americans participated in the study. Knowledge of prevention was improved after educational delivery. The average infection rate was 4.5%, with Cambodian, Thai, Vietnamese, Chinese and Korean groups having higher infection rates. The age group of 36-45 had the highest percentage of carriers (9.1%). CONCLUSION: Many Asian groups, particularly those of a southeast Asian decent, were subject to a higher probability of hepatitis-B infection. At an increased risk are first-generation Asian immigrants, groups with low immunization rates and those aged 36-45. The findings provide potential directions for focusing preventive interventions on at-risk Asian communities to reduce liver cancer disparities.

Evaluation of particle dispersal from mining and milling operations using lead isotopic fingerprinting techniques, Rio Pilcomayo Basin, Bolivia.

Mining and milling of ores from the Cerro Rico de Potosí precious metal-polymetallic tin deposits of Bolivia have led to severe contamination of water and sediments of the Rio Pilcomayo drainage system. Lead (Pb) isotopic data were used in this study to first document downstream dispersal patterns of Pb contaminated sediment within the channel of the Rio Pilcomayo, and then to determine the relative contribution of Pb from Cerro Rico within alluvial terrace soils that are used for agriculture. The concentration and isotopic composition of Pb within channel bed sediments differed significantly between 2000, 2002, and 2004. These differences presumably reflect changes in the type of ore mined and milled at Cerro Rico, and alterations in dispersal and grain-size dilution mechanisms associated with interannual variations in rainfall and runoff. Within agricultural terrace soils, both Pb concentrations and the percentage of Pb from Cerro Rico: (1) semi-systematically decrease downstream, (2) were found to decrease with terrace height above the channel, and (3) reflect the use of contaminated irrigation water. In upstream reaches (within 30 km of the mills), Pb from mining represents the most significant Pb source, accounting for more than 80% of Pb in the examined agricultural fields. At Sotomayor, located approximately 170 km from the mills, the relative contribution of Pb from Cerro Rico is highly variable between fields, but can be significant, ranging from approximately 15% to 35%. The analysis demonstrates that Pb isotopic ratios can be used to effectively trace contaminated particles through river systems and into adjacent alluvial soils, even where multiple Pb sources exist and Pb concentrations are similar to background values.

A spatial-temporal approach to surveillance of prostate cancer disparities in population subgroups.

BACKGROUND: Prostate cancer mortality disparities exist among racial/ethnic groups in the United States, yet few studies have explored the spatiotemporal trend of the disease burden. To better understand mortality disparities by geographic regions over time, the present study analyzed the geographic variations of prostate cancer mortality by three Texas racial/ethnic groups over a 22-year period. METHODS: The Spatial Scan Statistic developed by Kulldorff et al was used. Excess mortality was detected using scan windows of 50% and 90% of the study period and a spatial cluster size of 50% of the population at risk. Time trend was analyzed to examine the potential temporal effects of clustering. Spatial queries were used to identify regions with multiple racial/ethnic groups having excess mortality. RESULTS: The most likely area of excess mortality for blacks occurred in Dallas-Metroplex and upper east Texas areas between 1990 and 1999; for Hispanics, in central Texas between 1992 and 1996: and for non-Hispanic whites, in the upper south and west to central Texas areas between 1990 and 1996. Excess mortality persisted among all racial/ethnic groups in the identified counties. The second scan revealed that three counties in west Texas presented an excess mortality for Hispanics from 1980-2001. Many counties bore an excess mortality burden for multiple groups. There is no time trend decline in prostate cancer mortality for blacks and non-Hispanic whites in Texas. CONCLUSION: Disparities in prostate cancer mortality among racial/ethnic groups existed in Texas. Central Texas counties with excess mortality in multiple subgroups warrant further investigation.

Surveillance of the colorectal cancer disparities among demographic subgroups: a spatial analysis.

OBJECTIVE: The literature suggests that colorectal cancer mortality in Texas is distributed inhomogeneously among specific demographic subgroups and in certain geographic regions over an extended period. To understand the extent of the demographic and geographic disparities, the present study examined colorectal cancer mortality in 15 demographic groups in Texas counties between 1990 and 2001. METHODS: The Spatial Scan Statistic was used to assess the standardized mortality ratio, duration and age-adjusted rates of excess mortality, and their respective p-values for testing the null hypothesis of homogeneity of geographic and temporal distribution. RESULTS: The study confirmed the excess mortality in some Texas counties found in the literature, identified 13 additional excess mortality regions, and found 4 health regions with persistent excess mortality involving several population subgroups. CONCLUSION: Health disparities of colorectal cancer mortality continue to exist in Texas demographic subpopulations. Health education and intervention programs should be directed to the at-risk subpopulations in the identified regions.

Three non-allelic epistatically interacting methyltransferase mutations produce novel tocopherol (vitamin E) profiles in sunflower.

Wildtype sunflower (Helianthus annuus L.) seeds are a rich source of alpha-tocopherol (vitamin E). The g = Tph(2) mutation disrupts the synthesis of alpha-tocopherol, enhances the synthesis of gamma-tocopherol, and was predicted to knock out a gamma-tocopherol methyltransferase (gamma-TMT) necessary for the synthesis of alpha-tocopherol in sunflower seeds--wildtype (g(+) g(+)) lines accumulated > 90% alpha-tocopherol, whereas mutant (g g) lines accumulated > 90% gamma-tocopherol. We identified and isolated two gamma-TMT paralogs (gamma-TMT-1 and gamma-TMT-2). Both mapped to linkage group 8, cosegregated with the g locus, and were transcribed in developing seeds of wildtype lines. The g mutation greatly decreased gamma-TMT-1 transcription, caused alternative splicing of gamma-TMT-1, disrupted gamma-TMT-2 transcription, and knocked out one of two transcription initiation sites identified in the wildtype; gamma-TMT transcription was 36 to 51-fold greater in developing seeds of wildtype (g(+) g(+)) than mutant (g g) lines. F(2) populations (B109 x LG24 and R112 x LG24) developed for mapping the g locus segregated for a previously unidentified locus (d). B109, R112, and LG24 were homozygous for a null mutation (m = Tph(1)) in MT-1, one of two 2-methyl-6-phytyl-1,4-benzoquinone/2-methyl-6-solanyl-1,4-benzoquinone methyltransferase (MPBQ/MSBQ-MT) paralogs identified in sunflower. The d mutations segregating in B109 x LG24 and R112 x LG24 were allelic to a cryptic mutation identified in the other MPBQ/MSBQ-MT paralog (MT-2) and disrupted the synthesis of alpha- and gamma-tocopherol in F(2) progeny carrying m or g mutations--m m g(+) g(+) d d homozygotes accumulated 41.5% alpha- and 58.5% beta-T, whereas m m g g d d homozygotes accumulated 58.1% gamma- and 41.9% delta-T. MT-2 cosegregated with d and mapped to linkage group 4. Hence, novel tocopherol profiles are produced in sunflower seed oil by three non-allelic epistatically interacting methyltransferase mutations.