Chronic convection-enhanced delivery of topotecan for patients with recurrent glioblastoma: a first-in-patient, single-centre, single-arm, phase 1b trial.

BACKGROUND: Topotecan is cytotoxic to glioma cells but is clinically ineffective because of drug delivery limitations. Systemic delivery is limited by toxicity and insufficient brain penetrance, and, to date, convection-enhanced delivery (CED) has been restricted to a single treatment of restricted duration. To address this problem, we engineered a subcutaneously implanted catheter-pump system capable of repeated, chronic (prolonged, pulsatile) CED of topotecan into the brain and tested its safety and biological effects in patients with recurrent glioblastoma. METHODS: We did a single-centre, open-label, single-arm, phase 1b clinical trial at Columbia University Irving Medical Center (New York, NY, USA). Eligible patients were at least 18 years of age with solitary, histologically confirmed recurrent glioblastoma showing radiographic progression after surgery, radiotherapy, and chemotherapy, and a Karnofsky Performance Status of at least 70. Five patients had catheters stereotactically implanted into the glioma-infiltrated peritumoural brain and connected to subcutaneously implanted pumps that infused 146 µM topotecan 200 µL/h for 48 h, followed by a 5-7-day washout period before the next infusion, with four total infusions. After the fourth infusion, the pump was removed and the tumour was resected. The primary endpoint of the study was safety of the treatment regimen as defined by presence of serious adverse events. Analyses were done in all treated patients. The trial is closed, and is registered with ClinicalTrials.gov, NCT03154996. FINDINGS: Between Jan 22, 2018, and July 8, 2019, chronic CED of topotecan was successfully completed safely in all five patients, and was well tolerated without substantial complications. The only grade 3 adverse event related to treatment was intraoperative supplemental motor area syndrome (one [20%] of five patients in the treatment group), and there were no grade 4 adverse events. Other serious adverse events were related to surgical resection and not the study treatment. Median follow-up was 12 months (IQR 10-17) from pump explant. Post-treatment tissue analysis showed that topotecan significantly reduced proliferating tumour cells in all five patients. INTERPRETATION: In this small patient cohort, we showed that chronic CED of topotecan is a potentially safe and active therapy for recurrent glioblastoma. Our analysis provided a unique tissue-based assessment of treatment response without the need for large patient numbers. This novel delivery of topotecan overcomes limitations in delivery and treatment response assessment for patients with glioblastoma and could be applicable for other anti-glioma drugs or other CNS diseases. Further studies are warranted to determine the effect of this drug delivery approach on clinical outcomes. FUNDING: US National Institutes of Health, The William Rhodes and Louise Tilzer Rhodes Center for Glioblastoma, the Michael Weiner Glioblastoma Research Into Treatment Fund, the Gary and Yael Fegel Foundation, and The Khatib Foundation.

Interventions to prevent spontaneous preterm birth in women with singleton pregnancy who are at high risk: systematic review and network meta-analysis.

OBJECTIVES: To compare the efficacy of bed rest, cervical cerclage (McDonald, Shirodkar, or unspecified type of cerclage), cervical pessary, fish oils or omega fatty acids, nutritional supplements (zinc), progesterone (intramuscular, oral, or vaginal), prophylactic antibiotics, prophylactic tocolytics, combinations of interventions, placebo or no treatment (control) to prevent spontaneous preterm birth in women with a singleton pregnancy and a history of spontaneous preterm birth or short cervical length. DESIGN: Systematic review with bayesian network meta-analysis. DATA SOURCES: The Cochrane Pregnancy and Childbirth Group's Database of Trials, the Cochrane Central Register of Controlled Trials, Medline, Embase, CINAHL, relevant journals, conference proceedings, and registries of ongoing trials. ELIGIBILITY CRITERIA FOR SELECTING STUDIES: Randomised controlled trials of pregnant women who are at high risk of spontaneous preterm birth because of a history of spontaneous preterm birth or short cervical length. No language or date restrictions were applied. OUTCOMES: Seven maternal outcomes and 11 fetal outcomes were analysed in line with published core outcomes for preterm birth research. Relative treatment effects (odds ratios and 95% credible intervals) and certainty of evidence are presented for outcomes of preterm birth <34 weeks and perinatal death. RESULTS: Sixty one trials (17 273 pregnant women) contributed data for the analysis of at least one outcome. For preterm birth <34 weeks (40 trials, 13 310 pregnant women) and with placebo or no treatment as the comparator, vaginal progesterone was associated with fewer women with preterm birth <34 weeks (odds ratio 0.50, 95% credible interval 0.34 to 0.70, high certainty of evidence). Shirodkar cerclage showed the largest effect size (0.06, 0.00 to 0.84), but the certainty of evidence was low. 17OHPC (17α-hydroxyprogesterone caproate; 0.68, 0.43 to 1.02, moderate certainty), vaginal pessary (0.65, 0.39 to 1.08, moderate certainty), and fish oil or omega 3 (0.30, 0.06 to 1.23, moderate certainty) might also reduce preterm birth <34 weeks compared with placebo or no treatment. For the fetal outcome of perinatal death (30 trials, 12 119 pregnant women) and with placebo or no treatment as the comparator, vaginal progesterone was the only treatment that showed clear evidence of benefit for this outcome (0.66, 0.44 to 0.97, moderate certainty). 17OHPC (0.78, 0.50 to 1.21, moderate certainty), McDonald cerclage (0.59, 0.33 to 1.03, moderate certainty), and unspecified cerclage (0.77, 0.53 to 1.11, moderate certainty) might reduce perinatal death rates, but credible intervals could not exclude the possibility of harm. Only progesterone treatments are associated with reduction in neonatal respiratory distress syndrome, neonatal sepsis, necrotising enterocolitis, and admission to neonatal intensive care unit compared with controls. CONCLUSION: Vaginal progesterone should be considered the preventative treatment of choice for women with singleton pregnancy identified to be at risk of spontaneous preterm birth because of a history of spontaneous preterm birth or short cervical length. Future randomised controlled trials should use vaginal progesterone as a comparator to identify better treatments or combination treatments. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42020169006.

Fall Tillage Reduced Nutrient Loads from Liquid Manure Application during the Freezing Season.

Reducing agricultural runoff is important year round, particularly on landscapes that receive wintertime applications of manure. No-tillage systems are typically associated with reduced runoff loads during the growing season, but surface roughness from fall tillage may aid infiltration on frozen soils by providing surface depressional storage. The timing of winter manure applications may also affect runoff, depending on snow and soil frost conditions. Therefore, the objective of this study was to evaluate runoff and nutrient loads during the freezing season from combinations of tillage and manure application timings. Six management treatments were tested in south-central Wisconsin during the winters of 2015-2016 and 2016-2017 with a complete factorial design: two tillage treatments (fall chisel plow vs. no-tillage) and three manure application timings (early December, late January, and unmanured). Nutrient loads from winter manure application were lower on chisel-plowed versus untilled soils during both monitoring years. Loads were also lower from manure applied to soils with less frost development. Wintertime manure applications pose a risk of surface nutrient losses, but fall tillage and timing applications to thawed soils can help reduce loads.

Dynamics of Measured and Simulated Dissolved Phosphorus in Runoff from Winter-Applied Dairy Manure.

Agricultural P loss from fields is an issue due to water quality degradation. Better information is needed on the P loss in runoff from dairy manure applied in winter and the ability to reliably simulate P loss by computer models. We monitored P in runoff during two winters from chisel-tilled and no-till field plots that had liquid dairy manure applied in December or January. Runoff total P was dominated by nondissolved forms when soils were bare and unfrozen. Runoff from snow-covered, frozen soils had much less sediment and sediment-related P, and much more dissolved P. Transport of manure solids was greatest when manure was applied on top of snow and runoff shortly after application was caused by snowmelt. Dissolved P concentrations in runoff were greater when manure was applied on top of snow because manure liquid remained in the snowpack and allowed more P to be available for loss. Dissolved runoff P also increased as the amount of rain or snowmelt that became runoff (runoff ratio) increased. The SurPhos manure P runoff model reliably simulated these processes to provide realistic predictions of dissolved P in runoff from surface manure. Overall, for liquid dairy manure applied in winter, dissolved P concentrations in runoff can be decreased if manure is applied onto bare, unfrozen soil, or if runoff ratio can be reduced, perhaps through greater soil surface roughness from fall tillage. Both management approaches will allow more manure P to infiltrate into soil and less move in runoff. SurPhos is a tool that can reliably evaluate P loss for different management and policy scenarios for winter manure application.

Seasonal Variation in Sediment and Phosphorus Yields in Four Wisconsin Agricultural Watersheds.

Agricultural water quality projects in two distinct topographic regions in Wisconsin collected 5 to 10 yr of continuous stream discharge, suspended sediment (SS), total P (TP), and total dissolved P (TDP) in four watersheds (2100-5000 ha) from 2006 to 2016. Previous agricultural nonpoint SS and TP reduction efforts in two of these watersheds documented cold versus warm season differences in water quality response. The goal of this study was to identify seasonal partitioning of SS, TP, and TDP in storm event loads to inform stream water quality protection efforts. We used National Weather Service Coop Observer frost depth reports to identify dates when watershed soils were frozen. By comparing daily mean event discharge for dates relative to frost, we identified a 32-d post-frost high-discharge "thaw" period. Combined, the frozen and thaw periods contributed about half of the annual SS and TP runoff event loads, ranging from 47 to 63% for SS and from 45 to 51% for TP. The proportion of runoff event TDP during this time was even higher, 62 to 79%, with the majority during thaw. Watershed average volumetric runoff coefficients (event flow/precipitation and snowmelt) were two to four times higher during the freeze and the thaw compared with the rest of the year. To reduce total stream loads in regions with similar climates to Wisconsin, this study indicates that using management practices that curb sediment and P delivery to streams in the winter and early spring may be as important as those designed for nonfrozen conditions.

Temperature and Manure Placement in a Snowpack Affect Nutrient Release from Dairy Manure during Snowmelt.

Agricultural nutrient management is an issue due to N and P losses from fields and water quality degradation. Better information is needed on the risk of nutrient loss in runoff from dairy manure applied in winter. We investigated the effect of temperature on nutrient release from liquid and semisolid manure to water, and of manure quantity and placement within a snowpack on nutrient release to melting snow. Temperature did not affect manure P and NH-N release during water extraction. Manure P release, but not NH-N release, was significantly influenced by the water/manure solids extraction ratio. During snowmelt, manure P release was not significantly affected by manure placement in the snowpack, and the rate of P release decreased as application rate increased. Water extraction data can reliably estimate P release from manure during snowmelt; however, snowmelt water interaction with manure of greater solids content and subsequent P release appears incomplete compared with liquid manures. Manure NH-N released during snowmelt was statistically the same regardless of application rate. For the semisolid manure, NH-N released during snowmelt increased with the depth of snow covering it, most likely due to reduced NH volatilization. For the liquid manure, there was no effect of manure placement within the snowpack on NH-N released during snowmelt. Water extraction data can also reliably estimate manure NH-N release during snowmelt as long as NH volatilization is accounted for with liquid manures for all placements in a snowpack and semisolid manures applied on top of snow.

Spatially explicit methodology for coordinated manure management in shared watersheds.

Increased clustering and consolidation of livestock production systems has been linked to adverse impacts on water quality. This study presents a methodology to optimize manure management within a hydrologic region to minimize agricultural phosphorus (P) loss associated with winter manure application. Spatial and non-spatial data representing livestock, crop, soil, terrain and hydrography were compiled to determine manure P production rates, crop P uptake, existing manure storage capabilities, and transportation distances. Field slope, hydrologic soil group (HSG), and proximity to waterbodies were used to classify crop fields according to their runoff risk for winter-applied manure. We use these data to construct a comprehensive optimization model that identifies optimal location, size, and transportation strategy to achieve environmental and economic goals. The environmental goal was the minimization of daily hauling of manure to environmentally sensitive crop fields, i.e., those classified as high P-loss fields, whereas the economic goal was the minimization of the transportation costs across the entire study area. A case study encompassing two contiguous 10-digit hydrologic unit subwatersheds (HUC-10) in South Central Wisconsin, USA was developed to demonstrate the proposed methodology. Additionally, scenarios representing different management decisions (storage facility maximum volume, and project capital) and production conditions (increased milk production and 20-year future projection) were analyzed to determine their impact on optimal decisions.

Quantifying the Impact of Seasonal and Short-term Manure Application Decisions on Phosphorus Loss in Surface Runoff.

Agricultural phosphorus (P) management is a research and policy issue due to P loss from fields and water quality degradation. Better information is needed on the risk of P loss from dairy manure applied in winter or when runoff is imminent. We used the SurPhos computer model and 108 site-years of weather and runoff data to assess the impact of these two practices on dissolved P loss. Model results showed that winter manure application can increase P loss by 2.5 to 3.6 times compared with non-winter applications, with the amount increasing as the average runoff from a field increases. Increased P loss is true for manure applied any time from late November through early March, with a maximum P loss from application in late January and early February. Shifting manure application to fields with less runoff can reduce P loss by 3.4 to 7.5 times. Delaying manure application when runoff is imminent can reduce P loss any time of the year, and sometimes quite significantly, but the number of times that application delays will reduce P loss is limited to only 3 to 9% of possible spreading days, and average P loss may be reduced by only 15% for winter-applied manure and 6% for non-winter-applied manure. Overall, long-term strategies of shifting manure applications to low runoff seasons and fields can potentially reduce dissolved P loss in runoff much more compared with near-term, tactical application decisions of avoiding manure application when runoff is imminent.

OUTCOME OF IMPLEMENTATION OF A MULTIDISCIPLINARY TEAM APPROACH TO THE CARE OF PATIENTS AFTER TRANSSPHENOIDAL SURGERY.

OBJECTIVE: Transsphenoidal surgery (TS) for sellar lesions is an established and safe procedure, but complications can occur, particularly involving the neuroendocrine system. We hypothesized that postoperative care of TS patients would be optimized when performed by a coordinated team including a pituitary neurosurgeon, endocrinologists, and a specialty nurse. METHODS: We implemented a formalized, multidisciplinary team approach and standardized postoperative protocols for the care of adult patients undergoing TS by a single surgeon (J.N.B.) at our institution beginning in July 2009. We retrospectively compared the outcomes of 214 consecutive TS-treated cases: 113 cases prior to and 101 following the initiation of the team approach and protocol implementation. Outcomes assessed included the incidence of neurosurgical and endocrine complications, length of stay (LOS), and rates of hospital readmission and unscheduled clinical visits. RESULTS: The median LOS decreased from 3 days preteam to 2 days postteam (P<.01). Discharge occurred on postoperative day 2 in 46% of the preteam group patients compared to 69% of the postteam group (P<.01). Rates of early postoperative diabetes insipidus (DI) and readmissions within 30 days for syndrome of inappropriate antidiuretic hormone (SIADH) or other complications did not differ between groups. CONCLUSION: Implementation of a multidisciplinary team approach was associated with a reduction of LOS. Despite earlier discharge, postoperative outcomes were not compromised. The endocrinologist is central to the success of this team approach, which could be successfully applied to care of patients undergoing TS, as well as other types of endocrine surgery at other centers.

Phosphorus indices: why we need to take stock of how we are doing.

Many states have invested significant resources to identify components of their Phosphorus (P) Index that reliably estimate the relative risk of P loss and incentivize conservation management. However, differences in management recommendations and manure application guidelines for similar field conditions among state P Indices, coupled with minimal reductions in the extent of P-impaired surface waters and soil test P (STP) levels, led the U.S. Natural Resources Conservation Service (NRCS) to revise the 590 Nutrient Management Standard. In preparation for this revision, NRCS requested that a review of the scientific underpinnings and accuracy of current P Indices be undertaken. They also sought to standardize the interpretation and management implications of P Indices, including establishment of ratings above which P applications should be curtailed. Although some states have initiated STP thresholds above which no application of P is allowed, STP alone cannot define a site's risk of P loss. Phosphorus Indices are intended to account for all of the major factors leading to P loss. A rigorous evaluation of P Indices is needed to determine if they are directionally and magnitudinally correct. Although use of observed P loss data under various management scenarios is ideal, such data are spatially and temporally limited. Alternatively, the use of a locally validated water quality model that has been shown to provide accurate estimates of P loss may be the most expedient option to conduct Index assessments in the short time required by the newly revised 590 Standard.

Testing the Wisconsin Phosphorus Index with year-round, field-scale runoff monitoring.

The Wisconsin Phosphorus Index (WPI) is one of several P indices in the United States that use equations to describe actual P loss processes. Although for nutrient management planning the WPI is reported as a dimensionless whole number, it is calculated as average annual dissolved P (DP) and particulate P (PP) mass delivered per unit area. The WPI calculations use soil P concentration, applied manure and fertilizer P, and estimates of average annual erosion and average annual runoff. We compared WPI estimated P losses to annual P loads measured in surface runoff from 86 field-years on crop fields and pastures. As the erosion and runoff generated by the weather in the monitoring years varied substantially from the average annual estimates used in the WPI, the WPI and measured loads were not well correlated. However, when measured runoff and erosion were used in the WPI field loss calculations, the WPI accurately estimated annual total P loads with a Nash-Sutcliffe Model Efficiency (NSE) of 0.87. The DP loss estimates were not as close to measured values (NSE = 0.40) as the PP loss estimates (NSE = 0.89). Some errors in estimating DP losses may be unavoidable due to uncertainties in estimating on-farm manure P application rates. The WPI is sensitive to field management that affects its erosion and runoff estimates. Provided that the WPI methods for estimating average annual erosion and runoff are accurately reflecting the effects of management, the WPI is an accurate field-level assessment tool for managing runoff P losses.

Growing leaders in a professional membership organization.

AIMS: This study describes the background, development and delivery of an innovative approach to prepare elected state association leaders for their new leadership roles in their respective state organizations. BACKGROUND: State-based professional associations face unprecedented threats to scope of practice in the current volatile healthcare environment. In 2009 the American Association of Nurse Anesthetists (AANA) began offering a Leadership Development 'Boot Camp' for all nurse anesthetist state association presidents-elect designed with the intent of providing the participants with information, resources and tools they need in their new role. METHODS: An organizational assessment examined the philosophy of and need for leadership development across the entire organizational structure. RESULTS: Recommendations from the assessment included the need for intensive leadership development at the state level. A 3-day intensive workshop was designed and implemented. Adult learning theory was used for its development. CONCLUSIONS: The success of this programme has lead to its implementation as part of leadership development of the AANA. IMPLICATIONS FOR NURSING MANAGEMENT: Nurse managers and elected volunteer leaders in state-based professional associations face similar challenges requiring sound leadership ability. Principles applied to the development of the AANA programme can be applied to leadership development for new nurse managers.

A population-based study of parkinsonism in an Amish community.

BACKGROUND: Parkinson's disease (PD) is a neurodegenerative disorder with unknown cause. Genetic mutations account for a minority of cases but the role of environmental factors is unclear. METHODS: We performed a population-based screening for PD in subjects in an Amish community over age 60. PD was diagnosed using standard clinical criteria and the Unified Parkinson Disease Rating Scale motor subsection 3 (UPDRS3). Community prevalence was calculated. We constructed a community pedigree and calculated kinship coefficients, a measure of relatedness between 2 subjects, for every pair of subjects in diagnostic categories: clinically definite PD, UPDRS3 score >9, Mini-Mental State Exam (MMSE) score <25, and normal. RESULTS: Of 262 eligible subjects, 213 agreed to participate, 15 had PD, 43 had MMSE <25, 73 had UPDRS3 >9. The prevalence of PD was 5,703/100,000 with increasing prevalence in every decade of age. Excluding first-degree relatives, normal subjects were more related to each other (0.0102, SD = 0.0266) than subjects with clinically definite PD (0.0054, SD = 0.0100; p = 0.00003), subjects with UPDRS >9 (0.0076, SD = 0.0155; p = 0.00001), and subjects with MMSE <25 (0.0090, SD = 0.0180; p = 0.00003). CONCLUSIONS: PD and parkinsonian signs are common in this population and the prevalence increases with age. The finding that subjects with PD were not more related than normal subjects suggests that environmental factors may contribute to the parkinsonian phenotype in this community.

The acute-phase protein serum amyloid A3 is expressed in the bovine mammary gland and plays a role in host defence.

The serum amyloid A protein is one of the major reactants in the acute-phase response. Using representational difference analysis comparing RNA from normal and involuting quarters of a dairy cow mammary gland, we found an mRNA encoding the SAA3 protein (M-SAA3). The M-SAA3 mRNA was localized to restricted populations of bovine mammary epithelial cells (MECs). It was expressed at a moderate level in late pregnancy, at a low level through lactation, was induced early in milk stasis, and expressed at high levels in most MECs during mid to late involution and inflammation/mastitis. The mature M-SAA3 peptide was expressed in Escherichia coli, antibodies made, and shown to have antibacterial activity against E. coli, Streptococcus uberis and Pseudomonas aeruginosa. These results suggest that the mammary SAA3 may have a role in protection of the mammary gland during remodelling and infection and possibly in the neonate gastrointestinal tract.

Validity and reliability of an occupational exposure questionnaire for parkinsonism in welders.

This study assessed the validity and test-retest reliability of a medical and occupational history questionnaire for workers performing welding in the shipyard industry. This self-report questionnaire was developed for an epidemiologic study of the risk of parkinsonism in welders. Validity participants recruited from three similar shipyards were asked to give consent for access to personnel files and complete the questionnaire. Responses on the questionnaire were compared with information extracted from personnel records. Reliability participants were recruited from the same shipyards and were asked to complete the questionnaire at two different times approximately 4 weeks apart. Percent agreement, kappa, intraclass correlation coefficient (ICC), and sensitivity and specificity were used as measures of validity and/or reliability. Personnel files were obtained for 101 of 143 participants (70%) in the validity study, and 56 of the 95 (58.9%) participants in the reliability study completed the retest of the questionnaire. Validity scores for items extracted from personnel files were high. Percent agreement for employment dates and job titles ranged from 83-100%, while ICC for start and stop dates ranged from 0.93-0.99. Sensitivity and specificity for current job title ranged from 0.5-1.0. Reliability scores for demographic, medical and health behavior items were mainly moderate or high, but ranged from 0.19 to 1.0. Most recent job/title items such as title, types of welding performed, and material used showed substantial to perfect agreement. Certain determinants of exposure such as days and hours per week exposed to welding fumes demonstrated mainly moderate agreement (kappa= 0.42-0.47, percent agreement 63-77%); however, mean days and hours reported did not differ between test and retest. The results of this study suggest that participants' self-report for job title and dates employed are valid compared with employer records. While kappa scores were low for some medical conditions and for caffeine consumption, high kappa scores for job title, dates worked, types of welding, and materials welded suggest participants generated reproducible answers important for occupational exposure assessment.

Characterization of directly transformed weedy Brassica rapa and introgressed B. rapa with Bt cry1Ac and gfp genes.

Crop to weed transgene flow, which could result in more competitive weed populations, is an agricultural biosafety concern. Crop Brassica napus to weedy Brassica rapa hybridization has been extensively characterized to better understand the transgene flow and its consequences. In this study, weedy accessions of B. rapa were transformed with Bacillus thuringiensis (Bt) cry1Ac- and green fluorescence protein (gfp)-coding transgenes using Agrobacterium to assess ecological performance of the wild biotype relative to introgressed hybrids in which the transgenic parent was the crop. Regenerated transgenic B. rapa events were characterized by progeny analysis, Bt protein enzyme-linked immunosorbent assay (ELISA), Southern blot analysis, and GFP expression assay. GFP expression level and Bt protein concentration were significantly different between independent transgenic B. rapa events. Similar reproductive productivity was observed in comparison between transgenic B. rapa events and B. rapa x B. napus introgressed hybrids in greenhouse and field experiments. In the greenhouse, Bt transgenic plants experienced significantly less herbivory damage from the diamondback moth (Plutella xylostella). No differences were found in the field experiment under ambient, low, herbivore pressure. Directly transformed transgenic B. rapa plants should be a helpful experimental control to better understand crop genetic load in introgressed transgenic weeds.

Transformation and segregation of GFP fluorescence and glyphosate resistance in horseweed (Conyza canadensis) hybrids.

The goal of this research was to generate a breeding population of horseweed segregating for glyphosate resistance. In order to generate a marker to select between hybrids of glyphosate resistant (GR) and glyphosate susceptible (GS) horseweed, a GR horseweed accession from western Tennessee was transformed with a green fluorescent protein (GFP) transgene. The GFP marker allowed for the simple and accurate determination of GR hybrid plants by visual observation. GR plants were shown to be transgenic via the green fluorescence under UV light, and resistant to glyphosate when sprayed with the field-use-rate 0.84 kg acid equivalent ha(-1) of glyphosate (i.e. Roundup) herbicide. An in vitro screen for glyphosate resistance in seedlings was developed, and a 5 microM glyphosate concentration was found to reduce dry weight in GS seedlings but not in GR seedlings. The GR plants containing GFP were then hand-crossed with GS plants from eastern Tennessee under greenhouse conditions, with GS plants acting as the pollen acceptor. Resulting seed was collected and germinated for GFP fluorescence screening. Seedlings that exhibited the transgenic GFP phenotype were selected as F(1) hybrids between GR and GS horseweed. Thirty GSxGR hybrids were produced on the basis of a green-fluorescent GFP phenotype of GR plants. GSxGFP/GR F(1) hybrids produced F(2) seeds, and F(2) plants were shown to segregate for GFP fluorescence and glyphosate resistance independently. Both traits segregated at a Mendelian 3:1 ratio, indicating a single gene is responsible for each phenotype.

[18F]FDOPA PET as an endophenotype for Parkinson's Disease linkage studies.

Parkinson disease (PD) is a late onset disorder with age-dependent penetrance that may confound genetic studies, since affected individuals may not demonstrate clinical manifestations at the time of evaluation. The use of endophenotypes, biologic surrogates for clinical disease diagnoses, may permit more accurate classification of at-risk subjects. Positron emission tomography (PET) measurements of 6-[18F]fluorodopa ([18F]FDOPA) uptake indicate nigrostriatal neuronal integrity and may provide a useful endophenotype for PD linkage studies. We performed [18F]FDOPA PET in 11 members of a large, multi-incident Amish family with PD, 24 normals and 48 people with clinically definite idiopathic PD (PD controls). Clinical diagnoses in the Amish were clinically definite PD in four, clinically probable in one, clinically possible in five, and normal in one. Abnormal [18F]FDOPA posterior putamen uptake was defined as less than 3 standard deviations below the normal mean. The criteria were applied to the Amish sample to determine a PET endophenotype for each. We performed genetic simulations using SLINK to model the effect phenoconversion with the PET endophenotype had on logarithm of odds (LOD) scores. PET endophenotype confirmed the status of two clinically definite subjects. Two clinically definite Amish PD subjects had normal PETs. Two possible PD were converted to "PET definite PD." The remainder had normal PETs. The average maximum LOD score with the pre-PET was 6.14 +/- 0.84. Simulating phenoconversion of subjects with unknown phenotypes increased the LOD score to 7.36 +/- 1.23. The [18F]FDOPA PET endophenotype permits phenoconversion in multi-incident PD families and may increase LOD score accuracy and power of an informative pedigree.

A rapid method for mass screening for parkinsonism.

Epidemiology studies of parkinsonism employ a variety of techniques for unbiased sampling of populations. No current method permits mass screening of all subjects in a population for parkinsonism by movement disorders specialists. We developed and piloted a new approach to facilitate accurate and efficient screening of large populations for diagnosis of parkinsonism and provide data on sensitivity and specificity. We evaluated 2081 welders referred for medical-legal screening. Subjects were video taped using a standardized protocol, and videos were rated on the Unified Parkinson's Disease Rating Scale motor subsection 3 (UPDRS3). A "video rater" viewed video tapes and entered ratings through a web-based database. An "in-person" examiner performed a UPDRS3 examination in a randomly selected subgroup of 48 workers drawn from the 2081. We developed quantitative diagnostic criteria for parkinsonism that established minimum diagnostic thresholds based upon UPDRS3 scores and compared these criteria with diagnosis by an in-person examiner. Specificity of these criteria compared to in-person examination was 91-100% but sensitivity was 56%. A threshold UPDRS3 score greater than nine provided 100% sensitivity and 81% specificity. Liberal criteria identified 266 (13.1%) subjects with probable parkinsonism and 220 (10.8%) subjects with definite parkinsonism. Conservative criteria identified 260 (12.8%) with probable parkinsonism and 122 (6%) with definite parkinsonism. Our screening method permits rapid assessment of parkinsonian signs. An absolute UPDRS3 score greater than nine provided the best combination of sensitivity and specificity for the diagnosis of parkinsonism, while quantitative exam-based criteria for cardinal parkinsonian signs maximized specificity. Parkinsonism as diagnosed by our criteria was common in this group of welders.

Expression of a beta-defensin mRNA, lingual antimicrobial peptide, in bovine mammary epithelial tissue is induced by mastitis.

The expression of a beta-defensin, the lingual antimicrobial peptide (LAP), in response to mastitis was investigated by real-time PCR of RNA from mastitic and control udder quarters. There was a positive relationship between somatic cell count in milk and LAP expression. In situ hybridization showed that LAP mRNA was expressed in epithelial cells of mastitic tissue. These results suggest that LAP plays a role in the innate immune response to mastitis.