Genetics of phenological development and implications for seed yield in lentil.

Understanding phenology, its genetics and agronomic consequences, is critical for crop adaptation. Here we aim at (1) characterising lentil response to photoperiod with a focus on five loci: the lentil ELF3 ortholog Sn, two loci linked to clusters of lentil FT orthologs and two loci without candidates in chromosomes 2 and 5 (exp. 1: 36 lines, short and long day in phytotron); (2) establishing phenology-yield relationship (exp. 2: 25 lines, 11 field environments). A vintage perspective, where we quantify time trends in phenotype over three decades of breeding, links both experiments. Yield increased linearly from older to newer varieties at 29 kg ha-1 yr-1 or 1.5% yr-1, correlated negatively with flowering time in both winter- and summer-rainfall regimes, and decoupled from biomass in favourable environments. Time to flowering shortened from older to newer varieties at -0.56 % yr-1 in the field, and -0.42 % yr-1 (short day) and -0.99 % yr-1 (long day) in the phytotron. Early-flowering lines of diverse origin carried multiple early alleles for the five loci, indicating that at least some of these loci affect phenology additively. Current germplasm primarily features the early flowering haplotype for an FTb cluster region, hence the potential to increase phenological diversity with yield implications.

Child health, infant formula funding and South African health professionals: Eliminating conflict of interest.

Letters by Van Niekerk and Khan on article by Lake et al. (Lake L, Kroon M, Sanders D, et al. Child health, infant formula funding and South African health professionals: Eliminating conflict of interest. S Afr Med J 2019;109(12):902-906. https://doi.org/10.7196/SAMJ.2019.v109i12.14336); and response by Lake et al.

Child health, infant formula funding and South African health professionals: Eliminating conflict of interest.

Despite clear evidence of the benefits of exclusive and continued breastfeeding for children, women and society, far too few children in South Africa (SA) are breastfed. One of the major impediments to improving this situation is the continued and aggressive marketing of breastmilk substitutes (BMSs) and infiltration of the BMS industry into contexts with exposure to health professionals. In this article we, as academics, practitioners and child health advocates, describe contraventions of the regulations that protect breastfeeding in SA and argue that bold, proactive leadership to eliminate conflict of interest in respect of the BMS industry is urgently required, together with far greater investments in proven interventions to promote and support breastfeeding.

Application of a coproantigen ELISA as an indicator of efficacy against multiple life stages of Fasciola hepatica infections in sheep.

At present diagnosis of true resistance and determination of drug efficacy in Fasciola hepatica infection rely solely on terminal experiments. The coproantigen ELISA (cELISA) has been reported previously as a sensitive and specific tool appropriate to detect treatment failure, and potentially drug resistance. Two studies were conducted to determine whether the cELISA was appropriate for on-farm efficacy and resistance testing in Australian Merino sheep. In Study 1 sheep were infected orally with 50 F. hepatica metacercariae on three occasions, twelve, six and two weeks prior to a single flukicide treatment with triclabendazole, closantel or albendazole. Sheep were sampled weekly for a further seven weeks prior to necropsy. Following effective treatment, no faecal antigen was detected from 1 week. When immature stages (≤6 weeks) survived treatment, coproantigen reappeared from 6 weeks post-treatment. Therefore, cELISA conducted 1-4 weeks after treatment will demonstrate obvious treatment failure against adult F. hepatica, but is not sufficiently sensitive to detect survival of immature fluke until these reach maturity. In study 2, fluke burdens of sheep necropsied 13 weeks post single infection were compared to fecal worm egg counts (FWEC) and cELISA at necropsy. Regression analysis demonstrated that cELISA correlated strongly with fluke burden, whilst FWEC correlated weakly with cELISA. The correlation between FWEC and fluke burden was also weak, although stronger than that of FWEC with cELISA. The cELISA is an appropriate tool for monitoring effectiveness of treatments against Fasciola hepatica if an adult infection is present, however when immature stages of the parasite are present it is not as reliable. Where immature parasites are present it is recommended that initial cELISA be followed with a secondary cELISA at least 6 weeks after treatment to ensure resistance to immature stages is detected. Further testing is justified for monitoring the effectiveness of control programs by detecting adult populations that have survived a treatment regime.

Characterization of multiple life stages of two Australian Fasciola hepatica isolates in sheep.

Information on the susceptibility status of Fasciola hepatica isolates is lacking in the literature, even for those isolates considered to be laboratory reference strains. Four controlled efficacy studies were conducted on two Fasciola hepatica isolates from Australia, viz. 'Oberon' and 'Sunny Corner' with treatment at either 2, 6 or 10 weeks post-infection (wpi) as defined in each study. Fluke burdens and examination of livers occurred at necropsy in weeks 12 (Study 1) or 13 (Studies 2, 3 and 4) post-infection. The triclabendazole (TCBZ) resistance status of the Oberon isolate was confirmed in 6 and 10-week old F. hepatica, utilizing the drug alone (Fasinex; 71.5% and 31.1%, respectively) and in combination with oxfendazole (Flukazole C; 79.9% and 0%, respectively). The susceptibility of this isolate to albendazole, as well as salicylanilide and sulphonamide drugs was confirmed. The Sunny Corner isolate was confirmed as susceptible to TCBZ (>99% all stages) and closantel (>90% at ≥6 wpi).

Outbreak of tuberculosis in a homeless population involving multiple sites of transmission.

SETTING: During 2002-2003, a large outbreak of tuberculosis (TB) occurred among persons using multiple homeless facilities in King County, Washington. OBJECTIVE: To control the transmission of TB in multiple settings. DESIGN: In 2002, contacts exposed to patients in homeless facilities were screened using tuberculin skin tests (TSTs) and symptom review. Based on these screening results, sites of transmission were identified and prioritised, and exposed cohorts at these sites were offered intensive screening tests in 2003 (e.g., symptom review, TST, chest radiograph [CXR], sputum examination and culture). Mycobacterium tuberculosis isolates from patients were genotyped using PCR-based methods to identify outbreak-associated patients quickly. RESULTS: During 2002-2003, 48 (15%) of 313 patients diagnosed in King County were outbreak-associated; 47 culture-positive patients had isolates that matched the outbreak strain by genotyping. Three facilities visited by >12 patients in 2002 had a higher prevalence of TST positive results (approximately 30%) among clients compared with the background rate (7%) in the homeless community. Screening contacts with one sputum culture was as sensitive as CXR in detecting TB disease (77% vs. 62%, respectively). CONCLUSIONS: A comprehensive, resource-intensive approach likely helped to control transmission. This outbreak highlights the vulnerability of homeless populations and the need to maintain robust TB programs in urban settings.

Antileukemic properties of organoplatinum complexes.

The antitumor activity of 46 cis-amineplatinum congeners was evaluated against L1210 leukemia in (C57BL/L X DBA/2)F1 mice. Several compounds in this series significantly prolonged the life-spans of mice with the leukemia. During the selection of the compound that yielded optimal activity [dichloro(1,2-diaminocyclohexane)platinum], the chlorides were substituted with various organic and inorganic anions. The aqueous solubility was greatly increased with retention of significant antileukemic activity. Most of the active compounds were synergistic with cyclophosphamide, and cure rates up to 80% were obtained with certain combinations.

Toxicity and antitumor activity of hexamethylmelamine and its N-demethylated metabolites in mice with transplantable tumors.

N-Demethylated metabolites of the antineoplastic agent hexamethylmelamine were synthesized, and their toxicities and antitumor activities were determined in vivo. Determinations of the lethal dose for 10% of the male C57BL X DBA/2 F1 (hereafter called BD2F1) mice showed hexamethylmelamine toxicity to be decreased by N-demethylation; the metabolites showed a direct relationship between potency (mmoles/kg/day) and number of methyl groups present. In BD2F1 mice bearing Sarcoma 180 or Lewis lung carcinoma, the antitumor activities of the methylmelamines decreased with a reduction in number of methyl groups, but were similar at equitoxic levels. Results were similar in L1210 leukemic mice treated with lethal dose levels of the metabolites for 10% of the mice when mean survival times were measured. The therapeutic equality produced with equitoxic levels, together with the ineffectiveness of melamine, suggested that the presence of a methyl group, rather than the number, was the determining factor in the antitumor activity of the methylmelamines.

Effects of 17 beta-estradiol and progesterone on the metabolism of benz[a]pyrene by rabbit endometrial cells in primary culture.

The direct effects of estrogen (17 beta-estradiol) and progesterone on the metabolism of [3H]benzo[a]pyrene (BP) by primary cultures of rabbit endometrium epithelial cells were studied. 17 beta-Estradiol (10(-7) M) did not affect the metabolism of [3H]BP, expressed as the formation of water soluble metabolites of [3H]BP, during a 24-h incubation period. However, progesterone (10(-7) M) inhibited the formation of water-soluble metabolites of [3H]BP and the inhibitory effect was not prevented by 17 beta-estradiol. Progesterone did not appear to affect the relative distribution of the major conjugate types (sulfate esters and glutathiones) that comprised the water-soluble metabolites of [3H]BP. The results, which showed that progesterone acted to inhibit the metabolism of BP by endometrial cells, suggest additional evidence of a role for the ovarian hormone in the modulation of the carcinogenic efficacy of chemical carcinogens in a hormone-responsive tissue.

Regeneration of immobilized antibodies on fiber optic probes.

The regeneration of antibodies covalently immobilized to an optical fibre surface was investigated by dissociation of the antibody-antigen complex with three different solvents: (a) an acidic solution (0.1 M glycine hydrochloride in 50% (v/v) ethylene glycol, pH 1.75), (b) a basic solution (0.05 M tetraethylamine in 50% (v/v) ethylene glycol, pH 11.0) and (c) 50% (v/v) ethanol in PBS. The fibres coated with polyclonal rabbit anti-goat antibody against a large protein retained 70% and 65% of the original signal after five consecutive regenerations with acidic and basic solvent systems, respectively. The fibres coated with monoclonal mouse anti-trinitrobenzene antibody specific for a small organic molecule, retained over 90% of the original signal when regenerated with basic and ethanol solutions. This study evaluated regeneration and reuse of antibody-coated fibre optic biosensors as a means of reducing routine laboratory analysis costs and time.

Antibody immobilization using heterobifunctional crosslinkers.

Covalent attachment of functional proteins to a solid support is important for biosensors. One method employs thiol-terminal silanes and heterobifunctional crosslinkers such as N-succinimidyl 4-maleimidobutyrate (GMBS) to immobilize proteins through amino groups onto glass, silica, silicon or platinum surfaces. In this report, several heterobifunctional crosslinkers are compared to GMBS for their ability to immobilize active antibodies onto glass cover slips at a high density. Antibodies were immobilized at densities of 74-220 ng/cm2 with high levels of specific antigen binding. Carbohydrate-reactive crosslinkers were also compared to GMBS using a fiber optic biosensor to detect fluorescently-labeled antigen. At the concentrations tested, the antibodies immobilized with carbohydrate-reactive crosslinkers bound more antigen than GMBS immobilized antibodies as indicated by the fluorescence signal.

Effect of deoxyuridine coadministration on toxicity and antitumor activity of fluorouracil and floxuridine.

The addition of deoxyuridine (UDR) to fluorouracil (FU) or floxuridine (5-fluoro-2' deoxyuridine) (FUDR) produced a substantial increase in their toxicity in BDF1 mice. Antitumor assays using sarcoma 180 tumor-bearing mice showed a concomitant increase in tumor growth inhibition for the nucleoside-drug combination over identical doses of the single drug. However, no significant increase in antitumor activity with the combination treatment was demonstrated when equitoxic doses were given. Additional support for the therapeutic equality of the single and combination drug regimens was the similarity of the therapeutic indexes for each treatment regimen involving either fluorouracil or floxuridine. The results suggested that any therapeutic benefit achieved with the combination therapy could be duplicated with either fluorouracil or floxuridine at a higher dose.

Restrained eating: measuring an elusive construct.

As a measure of restrained eating, Herman's Restraint Scale (1978) reliably predicts laboratory food consumption in college students regardless of their weight. However, the generality and psychometric properties of the scale have not been established. In the present study, 136 male and female adults were cross-classified as obese and normal and as dieting or non-dieting. The subjects were administered a single questionnaire containing items of the Lie, Social Desirability, and Restraint scales presented in randomized order. Unlike previous reports by Herman, the three adult groups differed significantly on the Restraint Scale in the following order: Obese dieters greater than Obese non-dieters greater than normals. Also, alpha reliability coefficients varied across groups and corrected item-total correlations also displayed considerable variability with no uniformity apparent for individual item correlations. The factor analysis identified three factors within the ten item scale, and for the obese dieters, the scale was not independent of social desirability. These results indicate that the Restraint Scale has limited usefulness beyond laboratory settings with college students.

Explosives detection in soil using a field-portable continuous flow immunosensor.

A field method for quantitative analysis of explosives in contaminated soil samples is described. The method is based on a displacement immunoassay performed in a commercial instrument, the FAST 2000, engineered by Research International Inc. The method can be used on-site to measure 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) within 5min. For this study, replicate analyses were performed on soil extracts prepared from each field sample as well as appropriate controls, blanks, and laboratory standards. Statistical analyses were done to assess accuracy, bias, and predictability of the method. The results demonstrated that the immunosensor could be used effectively to screen environmental samples for the presence or absence of explosives. In most samples, the method also provided quantitative values that were in good agreement with standard laboratory analyses using HPLC. A limited number of sample matrices interfered with the immunoassay and produced results that varied significantly from the laboratory data. In each case, the compounds causing the problem have been identified and efforts are being made to minimize these matrix interferences in future field evaluations.

Establishing an audit system for Air Evac.

Because of JCAH requirements and nursing's progression toward self-evaluation, most fixed facilities have established audit systems to determine the effectiveness of nursing intervention. Due to the transient nature of the Aeromedical Evacuation System, there was no method of evaluating the patient's care inflight. Many obstacles had to be overcome in establishing a method of evaluation; and these problems are discussed by the researchers. A pilot project was formulated to determine the validity of our audit tool. Then, 99 patients' charts were audited over a four-month period. From this data, many deficiencies in the continuity of patient care and the patient's lack of understanding concerning our system became evident. The implications of this study are far reaching and will lead to further research in the future. By disseminating these results to various hospitals, we hope to see significant improvement in patient preparation in the near future.

Automated processing integrated with a microflow cytometer for pathogen detection in clinical matrices.

A spinning magnetic trap (MagTrap) for automated sample processing was integrated with a microflow cytometer capable of simultaneously detecting multiple targets to provide an automated sample-to-answer diagnosis in 40 min. After target capture on fluorescently coded magnetic microspheres, the magnetic trap automatically concentrated the fluorescently coded microspheres, separated the captured target from the sample matrix, and exposed the bound target sequentially to biotinylated tracer molecules and streptavidin-labeled phycoerythrin. The concentrated microspheres were then hydrodynamically focused in a microflow cytometer capable of 4-color analysis (two wavelengths for microsphere identification, one for light scatter to discriminate single microspheres and one for phycoerythrin bound to the target). A three-fold decrease in sample preparation time and an improved detection limit, independent of target preconcentration, was demonstrated for detection of Escherichia coli 0157:H7 using the MagTrap as compared to manual processing. Simultaneous analysis of positive and negative controls, along with the assay reagents specific for the target, was used to obtain dose-response curves, demonstrating the potential for quantification of pathogen load in buffer and serum.

Cellular and molecular toxicology of lead. III Effect of lead on heme synthesis.

The effect of lead nitrate on the biosynthesis of heme and the cellular uptake of (59)Fe was studied in an established rat liver cell line (RLC-GAI) grown in serum-free medium. Heme synthesis was shown to be increased in lead-treated cells when determined by measuring the amount of (59)Fe incorporated into the heme fraction of cell homogenates; concomitant with increased (59)Fe incorporation into heme, cellular uptake of (59)Fe was also increased. By contrast, heme synthesis was inhibited by lead in a dose-dependent manner when determined by measuring the conversion of ((14)C) aminolevulinic acid into heme under identical experimental conditions. Cellular degradation of heme was not found to be significantly affected by lead. It is proposed that lead inhibited the biosynthesis of heme and that the increased incorporation of (59)Fe into the heme fraction of the lead-treated cells was the result of the increased uptake of (59)Fe and hence labeling of the heme molecules. Evidence is presented that suggests that only (59)Fe present in solution as a low-molecular-weight complex was accumulated in increased amounts by lead-treated cells.