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Abstract There is a need for effective wound treatments that retain the bioactivity of a cellular treatment, but without the high costs and complexities associated with manufacturing, storing, and applying living biological products. Previously, we developed an amnion membrane‐derived hydrogel and evaluated its wound healing properties using a mouse wound model. In this study, we used a full thickness porcine skin wound model to evaluate the wound‐healing efficacy of the amnion hydrogel and a less‐processed amnion product comprising a lyophilized amnion membrane powder. These products were compared with commercially available amnion and nonamnion wound healing products. We found that the amnion hydrogel and amnion powder treatments demonstrated significant and rapid wound healing, driven primarily by new epithelialization versus closure by contraction. Histological analysis demonstrated that these treatments promote the formation of a mature epidermis and dermis with similar composition to healthy skin. The positive skin regenerative outcomes using amnion hydrogel and amnion powder treatments in a large animal model further demonstrate their potential translational value for human wound treatments.

Background: Outcomes for children with high-grade gliomas (HGG) remain poor. This multicenter phase II trial evaluated whether concurrent use of vorinostat or bevacizumab with focal radiotherapy (RT) improved 1-year event-free survival (EFS) compared to temozolomide in children with newly diagnosed HGG who received maintenance temozolomide and bevacizumab., Methods: Patients ≥ 3 and < 22 years with localized, non-brainstem HGG were randomized to receive RT (dose 54-59.4Gy) with vorinostat, temozolomide, or bevacizumab followed by 12 cycles of bevacizumab and temozolomide maintenance therapy., Results: Among 90 patients randomized, the 1-year EFS for concurrent bevacizumab, vorinostat, or temozolomide with RT was 43.8% (±8.8%), 41.4% (±9.2%), and 59.3% (±9.5%), respectively, with no significant difference among treatment arms. Three- and five-year EFS for the entire cohort was 14.8% and 13.4%, respectively, with no significant EFS difference among the chemoradiotherapy arms. IDH mutations were associated with more favorable EFS ( P  = .03), whereas H3.3 K27M mutations ( P  = .0045) and alterations in PIK3CA or PTEN ( P  = .025) were associated with worse outcomes. Patients with telomerase- and alternative lengthening of telomeres (ALT)-negative tumors ( n  = 4) had an EFS of 100%, significantly greater than those with ALT or telomerase, or both ( P  = .002). While there was no difference in outcomes based on TERT expression, high TERC expression was associated with inferior survival independent of the telomere maintenance mechanism ( P  = .0012)., Conclusions: Chemoradiotherapy with vorinostat or bevacizumab is not superior to temozolomide in children with newly diagnosed HGG. Patients with telomerase- and ALT-negative tumors had higher EFS suggesting that, if reproduced, mechanism of telomere maintenance should be considered in molecular-risk stratification in future studies., Competing Interests: None., (© The Author(s) 2024. Published by Oxford University Press, the Society for Neuro-Oncology and the European Association of Neuro-Oncology.)

Warm-season grasses (WSG) incorporated into traditional cool-season rotational grazing systems to increase summer yields are typically established in monoculture in separate pasture areas. Few studies have evaluated alternative interseeded establishment of WSG, despite potential benefits for improving biodiversity and land-use efficiency. The objective of this study was to determine the impact of establishment method (monoculture vs. interseeded) on crabgrass pasture forage yield, nutritive value, and preference under equine grazing. Three adult standardbred mares grazed two main plots on two consecutive days (8 hr/d) for three grazing events in 2019: Jul 28/29 (GRAZE 1), Aug 20/30 (GRAZE 2), Oct 1/2 (GRAZE 3). Each main plot contained four replicates of three treatments: mixed cool-season grass (CSG); Quick-N-Big crabgrass (CRB) [ Digitaria sanguinalis (L.) Scop.] interseeded into existing cool-season grass (INT), and CRB established as a monoculture (MON). The cool-season grass mix included Inavale orchardgrass [ Dactylis glomerata (L.)], Tower tall fescue [ Lolium arundinaceum (Schreb.) Darbysh.], and Argyle Kentucky bluegrass [ Poa pratensis (L.)]. Herbage mass (HM) and sward height (SH) were measured prior to each grazing event and samples were collected (0800-1000 h) for chemical composition analysis. Observed grazing time (GT) in each sub-plot as determined by 5-min scan sampling was utilized as marker of horse preference. Forage HM was greater in MON (8043 ± 1220 kg/ha) than CSG (5001 ± 1308 kg/ha; P = 0.003), with a trend for greater total HM in MON vs. INT (6582 ± 1220 kg/ha: P = 0.06), but HM did not differ between INT and CSG. The SH was also greatest for MON (28 ± 1.11; INT: 23.6 ± 1.11; CSG: 19.7 ± 1.37 cm; P < 0.003). Forage nutrients (digestible energy and crude protein) were largely similar across treatments and met requirements of horses at maintenance. Horse GT was lower in MON (22.6 ± 3.77 min/sub-plot) than in INT (31.9 ± 3.79 min/sub-plot; P = 0.003) and there was a trend for lower GT in MON vs. CSG (29.9 ± 4.17 min/sub-plot: P = 0.07). These results indicate interseeding CRB would not effectively increase yields of traditional cool-season grass equine rotational grazing systems and would not supply similar levels of summer forage provided by monoculture establishment. Results of this study also suggest horses may prefer cool-season grass pasture forage over warm-season crabgrass., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Integration of warm-season grasses into traditional cool-season pastures can increase summer forage for grazing cattle. The aim of this study was to determine impacts of this practice on yield and nutrient composition of equine rotational pasture systems as well as horse body condition. Two 1.5 ha rotational systems (6 to 0.25 ha sections/system) were evaluated: a control system (CON) (all sections mixed cool-season grass [CSG-CON]) and an integrated rotational grazing system (IRS) (three CSG sections [CSG-IRS] and three Quick-N-Big crabgrass [ Digitaria sanguinalis (L.) Scop.; CRB-IRS]). Three horses per system grazed in three periods: EARLY (mid-May to mid-July), SLUMP (mid-July to mid-September), and LATE (mid-September to mid-November). Herbage mass (HM) was measured prior to each rotation and samples were collected (0800 to 1000 h) for nutrient analysis. Grazing days were tracked to calculate carrying capacity (CC). Horse condition measures were assessed monthly. Over the full grazing season, 9,125 kg of forage was available for grazing in IRS versus 6,335 kg in CON. The CC was 390 horse d for IRS, while only 276 horse d for CON. Total HM/section did not differ during EARLY when CRB was not available (CSG-IRS: 2,537 ± 605; CSG-CON: 3,783 ± 856 kg/ha), but CC was greater in CSG-IRS (220 ± 37 horse d/ha) than CSG-CON (92 ± 26 horse d/ha; P = 0.03). In SLUMP, both HM and CC were greater in CRB-IRS (HM: 4,758 ± 698 kg/ha; CC: 196 ± 31 horse d/ha) than CSG-IRS (HM: 1,086 ± 698 kg/ha; CC: 32 ± 31 horse d/ha) or CON (HM: 970 ± 493 kg/ha; CC: 46 ± 22 horse d/ha; P < 0.02). While HM did not differ by section type in LATE (1,284 ± 158 kg/ha), CC was greater in CSG-CON (84 ± 9 horse d/ha) versus CRB-IRS (32 ± 13 horse d/ha; P = 0.03) and CSG-IRS (40 ± 13 horse d/ha; P = 0.06). During SLUMP, water-soluble carbohydrates (WSC) were lower in CRB-IRS (4.46% ± 0.80%) than CSG-CON (7.92% ± 0.90%; P < 0.04), but not CSG-IRS (5.93% ± 1.04%); however, non-structural carbohydrates (NSC) did not differ (7.05% ± 0.62%). There were no differences in WSC (6.46% ± 0.54%) or NSC (7.65% ± 0.54%) by section type in LATE. Horses in IRS maintained a body condition score (BCS) of 5.78 ± 0.48, but BCS did not differ by system (CON: 6.11 ± 0.48). Thus, integrated grazing increased summer pasture yield and provided adequate nutrition to maintain horse condition, but further research is needed to improve late-season production. Integrated grazing may not, however, provide an advantage in limiting dietary NSC, as NSC remained low for all pasture sections., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Charcoal rot caused by Macrophomina phaseolinais an increasing economic problem in annualized strawberry production systems around the world. Currently there are no effective postfumigation chemical controls for managing charcoal rot, and no information is available on the genetic architecture of resistance to M. phaseolina in strawberry (Fragaria ×ananassa). In this study, three multiparental discovery populations and two validation populations were inoculated at planting and evaluated for mortality in three consecutive growing seasons. Genome-wide SNP genotyping and pedigree-based analysis with FlexQTL™ software were performed. Two large-effect quantitative trait loci (QTL) increasing charcoal rot resistance were discovered and validated in cultivated germplasm. FaRMp1 was located on linkage group 2A in the interval 20.4to 24.9 cM, while FaRMp2 was located on linkage group 4B in the interval 41.1to 61.2 cM. Together these QTLs explained 27% and 17% of the phenotypic variance in two discovery populations consisting of elite breeding germplasm. For both QTLs, the resistant allele showed some evidence of partial dominance, but no significant interaction was detected between the two loci. As the dosage of resistant alleles increased from 0 to 4 across the two QTLs, mortality decreased regardless of the combination of alleles.A third locus, FaRMp3 on 4D, was discovered in FVC 11-58, a reconstituted F.×ananassa originating from diverse F. virginiana and F. chiloensis accessions. This locus accounted for 44% of phenotypic variation in four segregating crosses. These findings will form the basis for DNA-informed breeding for resistance to charcoal rot in cultivated strawberry., (© The Author(s) 2021. Published by Oxford University Press on behalf of Genetics Society of America.)

Aims: Despite the effects of statins in reducing cardiovascular events and slowing progression of coronary atherosclerosis, significant cardiovascular (CV) risk remains. Icosapent ethyl (IPE), a highly purified eicosapentaenoic acid ethyl ester, added to a statin was shown to reduce initial CV events by 25% and total CV events by 32% in the REDUCE-IT trial, with the mechanisms of benefit not yet fully explained. The EVAPORATE trial sought to determine whether IPE 4 g/day, as an adjunct to diet and statin therapy, would result in a greater change from baseline in plaque volume, measured by serial multidetector computed tomography (MDCT), than placebo in statin-treated patients., Methods and Results: A total of 80 patients were enrolled in this randomized, double-blind, placebo-controlled trial. Patients had to have coronary atherosclerosis as documented by MDCT (one or more angiographic stenoses with ≥20% narrowing), be on statin therapy, and have persistently elevated triglyceride (TG) levels. Patients underwent an interim scan at 9 months and a final scan at 18 months with coronary computed tomographic angiography. The pre-specified primary endpoint was change in low-attenuation plaque (LAP) volume at 18 months between IPE and placebo groups. Baseline demographics, vitals, and laboratory results were not significantly different between the IPE and placebo groups; the median TG level was 259.1 ± 78.1 mg/dL. There was a significant reduction in the primary endpoint as IPE reduced LAP plaque volume by 17%, while in the placebo group LAP plaque volume more than doubled (+109%) (P = 0.0061). There were significant differences in rates of progression between IPE and placebo at study end involving other plaque volumes including fibrous, and fibrofatty (FF) plaque volumes which regressed in the IPE group and progressed in the placebo group (P < 0.01 for all). When further adjusted for age, sex, diabetes status, hypertension, and baseline TG, plaque volume changes between groups remained significantly different, P < 0.01. Only dense calcium did not show a significant difference between groups in multivariable modelling (P = 0.053)., Conclusions: Icosapent ethyl demonstrated significant regression of LAP volume on MDCT compared with placebo over 18 months. EVAPORATE provides important mechanistic data on plaque characteristics that may have relevance to the REDUCE-IT results and clinical use of IPE., (© The Author(s) 2020. Published by Oxford University Press on behalf of the European Society of Cardiology.)

There is a need for effective wound treatments that retain the bioactivity of a cellular treatment, but without the high costs and complexities associated with manufacturing, storing, and applying living biological products. Previously, we developed an amnion membrane-derived hydrogel and evaluated its wound healing properties using a mouse wound model. In this study, we used a full thickness porcine skin wound model to evaluate the wound-healing efficacy of the amnion hydrogel and a less-processed amnion product comprising a lyophilized amnion membrane powder. These products were compared with commercially available amnion and nonamnion wound healing products. We found that the amnion hydrogel and amnion powder treatments demonstrated significant and rapid wound healing, driven primarily by new epithelialization versus closure by contraction. Histological analysis demonstrated that these treatments promote the formation of a mature epidermis and dermis with similar composition to healthy skin. The positive skin regenerative outcomes using amnion hydrogel and amnion powder treatments in a large animal model further demonstrate their potential translational value for human wound treatments., (© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.)

Background: Dyslipidemia plays a significant role in the progression of cardiovascular disease. The apolipoprotein (apo) A1 remnant ratio (apo A1/VLDL3-C + IDL-C) has recently been shown to be a strong predictor of death/myocardial infarction risk among women >50 years undergoing angiography. However, whether this ratio is associated with coronary heart disease risk among other populations is unknown. We evaluated the apo A1 remnant ratio and its components for coronary heart disease incidence., Design: Observational., Methods: Participants (N = 4722) of the Jackson Heart Study were evaluated. Baseline clinical characteristics and lipoprotein subfractions (Vertical Auto Profile method) were collected. Cox hazard regression analysis, adjusted by standard cardiovascular risk factors, was utilized to determine associations of lipoproteins with coronary heart disease., Results: Those with new-onset coronary heart disease were older, diabetic, smokers, had less education, used more lipid-lowering medication, and had a more atherogenic lipoprotein profile. After adjustment, the apo A1 remnant ratio (hazard ratio = 0.67 per 1-SD, p = 0.002) was strongly associated with coronary heart disease incidence. This association appears to be driven by the IDL-C denominator (hazard ratio = 1.23 per 1-SD, p = 0.007). Remnants (hazard ratio = 1.21 per 1-SD, p = 0.017), but not apo A1 (hazard ratio = 0.85 per 1-SD, p = 0.121) or VLDL3-C (hazard ratio = 1.13 per 1-SD, p = 0.120) were associated with coronary heart disease. Standard lipids were not associated with coronary heart disease incidence., Conclusion: We found the apo A1 remnant ratio to be strongly associated with coronary heart disease. This ratio appears to better stratify risk than standard lipids, apo A1, and remnants among a primary prevention cohort of African Americans. Its utility requires further study as a lipoprotein management target for risk reduction., (© The European Society of Cardiology 2015.)

Between birth and late adolescence, the human brain undergoes exponential maturational changes. Using in vivo magnetic resonance spectroscopy, we determined the developmental profile for 6 metabolites in 5 distinct brain regions based on spectra from 309 children from 0 to 18 years of age. The concentrations of N-acetyl-aspartate (an indicator for adult-type neurons and axons), creatine (energy metabolite), and glutamate (excitatory neurotransmitter) increased rapidly between birth and 3 months, a period of rapid axonal growth and synapse formation. Myo-inositol, implicated in cell signaling and a precursor of membrane phospholipid, as well as an osmolyte and astrocyte marker, declined rapidly during this period. Choline, a membrane metabolite and indicator for de novo myelin and cell membrane synthesis, peaked from birth until approximately 3 months, and then declined gradually, reaching a plateau at early childhood. Similarly, taurine, involved in neuronal excitability, synaptic potentiation, and osmoregulation, was high until approximately 3 months and thereafter declined. These data indicate that the first 3 months of postnatal life are a critical period of rapid metabolic changes in the development of the human brain. This study of the developmental profiles of the major brain metabolites provides essential baseline information for future analyses of the pediatric health and disease.

Progress in the development of effective therapies for diffuse intrinsic brainstem gliomas (DIBSGs) is compromised by the unavailability of tissue samples and the lack of noninvasive markers that can characterize disease status. The purpose of this study was to compare the metabolic profile of DIBSGs with that of astrocytomas elsewhere in the CNS and to determine whether the measurement of metabolic features can improve the assessment of disease status. Forty in vivo MR spectroscopy (MRS) studies of 16 patients with DIBSG at baseline and after radiation therapy were retrospectively reviewed. Control data for baseline studies of DIBSGs were obtained from 14 untreated regular and anaplastic astrocytomas. All spectra were acquired with single-voxel, short echo-time (35 ms), point-resolved spectroscopy. Absolute metabolite concentrations (mmol/kg) and lipid intensities (arbitrary units) were determined. At baseline, creatine and total choline (tCho) were significantly lower in DIBSGs than in astrocytomas elsewhere in the CNS (4.3 +/- 1.1 vs. 7.5 +/- 1.9 mmol/kg, p < 0.001; 1.9 +/- 0.7 vs. 4.2 +/- 2.6, p < 0.001). Serial MRS in individual subjects revealed increasing levels of tCho (p < 0.05) and lipids (p < 0.05) and reduced ratios of N-acetylaspartate, creatine, and myoinositol relative to tCho (all p < 0.01). Metabolic progression defined by increased tCho concentration in serial MRS preceded clinical deterioration by 2.4 +/- 2.7 months (p < 0.04). Low tCho of DIBSG at baseline is consistent with low proliferative tumors. Subsequent metabolic changes that have been associated with malignant degeneration preceded clinical deterioration. MRS provides early surrogate markers for disease progression.

Linezolid, an oxazolidinone antibiotic, inhibits bacterial protein synthesis by binding to 23S ribosomal RNA (rRNA). We studied 3 patients who experienced lactic acidosis while receiving linezolid therapy. The toxicity may have been caused by linezolid binding to mitochondrial 16S rRNA. Genetic polymorphisms may have contributed to the toxicity in 2 patients.

A series of charge-modified, dye-labeled 2', 3'-dideoxynucleoside-5'-triphosphates have been synthesized and evaluated as reagents for dye-terminator DNA sequencing. Unlike the commonly used dye-labeled terminators, these terminators possess a net positive charge and migrate in the opposite direction to dye-labeled Sanger fragments during electrophoresis. Post-sequencing reaction purification is not required to remove unreacted nucleotide or associated breakdown products prior to electrophoresis. Thus, DNA sequencing reaction mixtures can be loaded directly onto a separating medium such as a sequencing gel. The charge-modified nucleotides have also been shown to be more efficiently incorporated by a number of DNA polymerases than regular dye-labeled dideoxynucleotide terminators or indeed normal dideoxynucleoside-5'-triphosphates.

A novel series of charge-modified, dye-labeled 2',3'-dideoxynucleoside-triphosphate terminators were synthesized and evaluated as reagents for DNA sequencing. These terminators possess an advantage over existing reagents in that no purification is required to remove unreacted nucleotide or associated breakdown products prior to electrophoretic separation of the sequencing fragments. This obviates the need for a time consuming post-reaction work up, allowing direct loading of DNA sequencing reaction mixtures onto a slab gel. Thermo Sequenase II DNA polymerase poorly incorporates the charge-modified terminators compared with regular dye-labeled terminators. However, extending the linker arm between dye and nucleotide and using a mutant form of a related DNA polymerase can in part mitigate the decrease in substrate efficiency. We also present evidence that these charge-modified terminators can relieve gel compression artefacts when used with dGTP in sequencing reactions.

Objective: To determine the effects of human immunodeficiency virus (HIV) infection on children's development by identifying neurological and environmental variables associated with neuropsychological measures of cognitive development in HIV-seronegative (HIV-) and HIV-seropositive (HIV+)children and adolescents with hemophilia., Methods: Participants (N = 298; 60% HIV+) were males ages 7-19 years enrolled in the Hemophilia Growth and Development Study (HGDS). Least squares modeling was used to determine whether there was a difference at baseline in mean neuropsychological test scores by HIV status, age, and neurological baseline findings, adjusting for selected environmental and medical history variables., Results: The participants were within age expectations for general intelligence. Variables associated with lowered neuropsychological performance included academic problems, coordination and/or gait abnormalities, parents' education, and previous head trauma., Conclusions: Hemophilia-related morbidity has a subtle adverse influence on cognitive performance. HIV infection was not associated with neuropsychological dysfunction in this group even when MRI abnormalities were present.

Natural suppressor (NS) cells, which exert nonspecific suppressive activity in an unprimed manner, have been found in mouse, rabbit and monkey bone marrow (BM). In the present study, we characterize NS cells in human BM. NS activity was found in a fraction of low density (1.055-1.065 g/ml) BM cells that had been depleted of T cells, B cells, and monocytes. The NS activity was significantly decreased by the depletion of CD34+ or CD33+ cells but not CD56+ cells. The NS activity was indeed detected in isolated CD34+ cells and further enriched in CD34+CD33+ cells. Hematopoietic progenitor cells committed to the myeloid lineage were also enriched in the CD34+CD33+ cells, which significantly correlated to the NS activity. From these findings, it is strongly suggested that NS activity in human BM is exerted by the myeloid hematopoietic progenitors. Since cell-to-cell contact was not necessary for the action, NS cells seemed to secrete soluble mediator(s). Transforming growth factor-beta 1 and leukemia inhibitory factor were, however, not the candidates, based on experiments using neutralizing antibodies.

Maternal antibodies against the envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1) have previously been suggested to be important in influencing the rate of vertical transmission. In this study, serum antibody responses in mothers who did or did not transmit HIV-1 infection to their children were measured against the carboxy region of the transmembrane envelope glycoprotein gp41. Results indicate significantly higher binding reactivity of nontransmitter mothers compared with transmitters to three peptides spanning amino acids 771-810 and 841-856. In addition, high neutralization titers in maternal sera against HIV-1(MN) were associated with a nontransmission status. This is the initial report demonstrating a correlation between maternal antibody binding to epitopes within the carboxy region of gp41 envelope glycoprotein and lack of vertical transmission. Immunodetection that identifies antibodies to these regions in gp41 could therefore be considered a strategy to assess the risk of vertical transmission of HIV-1.

The Suppressor-mutator (Spm) family of maize transposable elements consists of autonomous Spm elements and nonautonomous defective Spm (dSpm) elements. One characteristic of this family is that the insertion of dSpm elements into a structural gene often permits some level of structural gene expression in the absence of SpM activity, and this structural gene expression is suppressed in trans by Spm activity. The Spm's subterminal repetitive regions (SRRs) contain several iterations of a 12-bp repeat motif. It had been proposed that binding of an Spm-encoded protein to these repeat motifs blocks structural gene transcriptional readthrough, thus suppressing gene expression. The bz-m13 allele of the bronze 1 locus contains a 2.24-kb dSpm insertion in the second exon of a Bz allele. In the absence of Spm activity, bz-m13 displays substantial Bz expression, and this expression is fully suppressed by Spm. Four intra-dSpm deletion derivatives are described in which this Bz expression is only partially suppressed by Spm. Each of these derivatives retains at least 12 SRR repeat motifs. Thus the presence of these repeat motifs is not sufficient to guarantee complete suppression by Spm. Some other property such as secondary structure or element size must play a role.

The defective Suppressor-mutator (dSpm)-induced allele bronze1-mutable 13 (bz1-m13) and many of its derivative alleles are leaky mutants with measurable levels of flavonol O3-glucosyltransferase activity. This activity results from splicing at acceptor site-1, one of two cryptic 3' splice sites within the dSpm insertion in bz1-m13. In this study, splicing in bz1-m13 change-in-state (CS) alleles CS-3 and CS-64 was shown to be altered from bz1-m13; previous work found altered splicing in CS-9. CS-64 is a null allele and lacks the acceptor site-1-spliced transcript because this site is deleted. CS-3 and CS-9 had increased levels of the acceptor site-1 transcript relative to bz1-m13 and increased enzymic activities. A deletion in CS-9 altered splicing by eliminating acceptor site-2. Both acceptor sites were intact in CS-3, but a deletion removed most of a 275-bp GC-rich sequence in dSpm. This suggests that GC-rich sequences affect splicing and is consistent with models postulating a role for AU content in the splicing of plant introns. Splicing does not necessarily occur, however, at the junction of AU-rich intron sequences and GC-rich exon sequences.

Background: Malignancy of the breast is frequently diagnosed through fine needle aspiration. In the hands of a skilled aspirator and cytopathologist, this can be a highly accurate procedure., Purpose: This study was undertaken to evaluate whether sufficient residual cells in the bore of the needle could be harvested and analyzed efficiently by flow cytometry analysis. The goal was then to determine the value of routine flow cytometry as an adjunctive technology in the interpretation of breast fine needle aspirations., Methods: Cells were rinsed from the needles of 83 consecutive diagnostic fine needle aspirates after preliminary inspection had confirmed adequate material was obtained for cytopathology. Cells were washed, and nuclei prepared by detergent treatment. After ribonuclease treatment, DNA was stained with the fluorescent marker propidium iodide. DNA content per cell was determined by flow cytometry by measurement of right-angle fluorescence., Results: Less than 4% of the samples were rejected for inadequate cell numbers. Flow cytometry criteria for evidence of malignancy included the presence of a DNA aneuploid population or an elevated rate of proliferation (13% or higher) of a diploid population. Accuracy of flow cytometry was based on cytopathologic interpretation in all cases except two which were based on results of excisional biopsy. The sensitivity of the flow cytometry analysis was 76%; the specificity was 100%, with results from flow cytometry pivotal in the correct diagnoses for two patients whose cytopathologic results were equivocal. Analysis of histograms indicated acceptable coefficients of variation for all populations. Gating analysis indicated the suitability of the material for this type of study, with an average of 85% of the events selected, or "gated in." Low recoveries were associated with the presence of necrotic debris in the sample., Conclusion: Flow cytometry can be a valuable adjunctive technology, capable of providing the cytopathologist with additional information regarding the character of cells analyzed.

A mutant allele of the maize brittle-1 (bt1) locus, brittle-1-mutable (bt1-m), was shown genetically and molecularly to result from the insertion of a defective Suppressor-mutator (dSpm) transposable element. An Spm-hybridizing restriction enzyme fragment, which cosegregates with the bt1-m allele and is absent from wild-type revertants of bt1-m, was identified and cloned. Non-Spm portions of it were used as probes to identify wild-type (Bt1) cDNAs in an endosperm library. The 4.3-kb bt1-m genomic clone contains a 3.3-kb dSpm, which is inserted in an exon and is composed of Spm termini flanking non-Spm sequences. RNA gel blot analyses, using a cloned Bt1 cDNA probe, indicated that Bt1 mRNA is present in the endosperm of developing kernels and is absent from embryo or leaf tissues. Several transcripts are produced by bt1-m. The deduced translation product from a 1.7-kb Bt1 cDNA clone has an apparent plastid transit peptide at its amino terminus and sequence similarity to several mitochondrial inner-envelope translocator proteins, suggesting a possible role in amyloplast membrane transport.

A consecutive cohort of 87 infants (46 infants less than 37 weeks gestational age and 41 term infants greater than or equal to 37 weeks gestation) admitted to the Neonatal Intensive Care Unit (NICU) and a convenience cohort of 27 term well babies at the University of Nebraska Medical Center (Omaha, NE) were evaluated for plasma beta-endorphin (beta E) levels during the first 4 h after birth. Demographic data, maternal history, and respiratory status at the time of sampling as well as development of documented apneic episodes during the initial hospitalization were analyzed for all infants. All NICU infants had higher plasma beta E levels than the control infants. Premature infants had significantly higher neonatal plasma beta E levels than term infants in either the control or NICU groups, but the response was gender specific; premature males had higher plasma beta E than premature females (P = 0.008). Perinatal stress, including respiratory problems, was associated with the increase in plasma beta E, but prematurity and being male were significantly predictors of an elevated plasma beta E level. Immaturity in respiratory control, as evaluated by the development of documented apneic episodes during the infant's initial hospitalization, did not correlate with an elevated perinatal plasma beta E level.

The Dissociation (Ds) mutant, Bz-wm, of the maize bronze-1 (bz) locus conditions a leaky phenotype. Plants carrying this mutant allele synthesize a low amount of an altered Bz gene product, which leads to reduced anthocyanin pigmentation in the seed. The molecular analysis reported here shows that the Bz-wm mutant has a 406-bp Ds1 insertion located 63 bp 5' to the start of Bz transcription. Furthermore, the Bz-wm allele contains three additional base pairs within the second exon, relative to the wild-type Bz allele. These additional nucleotides are believed to be derived from the 8-bp target site duplication created by an Activator (Ac) element in a previous allele in the series. The biochemical and molecular analyses of Bz-wm and revertants of Bz-wm indicate that the three additional nucleotides are responsible for the altered enzyme stability, while the Ds1 element affects the steady-state level of Bz-specific protein and RNA. Since the two mutations present in the Bz-wm mutant were each caused by the action of the Ac-Ds transposable element system, these results provide new insights into the ways that transposable elements can modify maize gene expression.

The bz-m13 allele of the bronze-1 (bz) locus in maize contains a 2.2-kb defective Suppressor-mutator (dSpm) transposable element inserted in the second exon. We compared bz expression in bz-m13 and five derivatives in which the dSpm insertion had sustained deletions ranging from 2 to 1300 bp. Tissues homozygous for bz-m13 in the absence of Spm-s activity were found to contain from 5 to 13% of the enzymatic activity conditioned by a wild-type allele at the bz locus. Tissues homozygous for the deletion derivatives contained enzymatic activities ranging from less than 1% to 67%. These differences are closely correlated with the steady-state level of one of two alternatively spliced transcripts. In all alleles bz transcription proceeds through the dSpm insert. Subsequent RNA processing uses the donor site of the single bz intron and either one of two alternative acceptor splice sites (AS1 and AS2) located within the dSpm sequence. Use of the AS1 removes all but 2 bp of dSpm sequence and produces the 1.8-kb transcript whose level corresponds closely to the level of enzymatic activity. Use of AS2 produces a transcript which retains more than 600 bp of dSpm sequence. Those derivatives in which AS2 is either deleted or inactivated have substantially increased levels of both the 1.8-kb transcript and enzymatic activity. We therefore document one sequence of events which began with the insertion of a transposable element and resulted in novel and stable introns which retain element-derived sequence and which in certain cases permit substantial host-gene expression.

The relation between verticality perception and body balance as influenced by perceptual style, age, and sex was investigated in 100 subjects. Perceptual style was measured by the Rod and Frame Test. Body balance data were obtained by two electromechanical force plates. The results of the measurements of perceptual style indicated that the younger groups were more accurate in estimating verticality than the older groups and that men were more accurate than women. With regard to the body balance shift data, however, no significant differences relative to age, sex, or perceptual style were found. The rejection of the hypothesis concerning body balance shift was interpreted in terms of the dominant influence of proprioceptive cues over visual cues in maintaining body balance. In normal standing activities, as long as the neuromuscular system is intact, conflicting stimuli from the visual field appear to have little effect on modifying body balance.

The Ds-controlled allele, bz-m4 Derivative 6856 [bz-m4 D6856], is reported to have an altered temporal- and tissue-specific pattern of gene expression. We have cloned this allele and have characterized it at the molecular level. The mutation was caused by the insertion of a complex transposon-like structure 36 base pairs downstream from the Bz mRNA cap site. The insert is 6.7-kbp long. Ds elements, each approximately 2 kbp in length, are at both ends of the insert. The sequence between the Ds elements is a partial duplication of flanking sequences from the 3' end of the Bz gene. These data suggest that Ds initially inserted near the 3' end of the gene and mobilized adjacent sequences as it transposed.