Theligonum cynocrambe: Developmental morphology of a peculiar rubiaceous herb

The annual Mediterranean herbTheligonum cynocrambe shows a peculiar combination of morphological characters, e.g., switch from decussate to spiral phyllotaxis with 90–100° divergence, combined with a change from interpetiolar to lateral stipules, anemophily, lack of calyx, flowers often dimerous to trimerous, corolla fused in both male and female flowers, male flowers extra-axillary, with 2–19 stamens per flower, female flowers axillary, with inferior uniovulate ovary, basilateral style and perianth, nut-like fruits with elaiosome. In male flowers the androecium emerges as an (uneven) elliptical rim with a central depression. This common girdling primordium is divided up into several stamen primordia. In male flowers with low stamen number the stamen primordia may occupy the corners alternating with the corolla lobes. There are no epipetalous androecial primordia that secondarily divide into stamens. Male flowers occasionally show a hemispherical base that may be interpreted as remnant of the inferior ovary. In female flowers a ring primordium grows into a tube on which the petal lobes arise. The perianth and style become displaced adaxially by uneven growth of the inferior ovary. The ovary is basically bilocular. The lower region of the ovary is provided with a septum that is overtopped and hidden by the single curved ovule.Theligonum is referred to theRubiaceae-Rubioideae, with theAnthospermeae andPaederieae as most closely related tribes.     

Energy expenditure in relation to activity of lesser mouse deer (Tragulus javanicus)

Heat production (HP) of male and female mouse deer during eating, standing and sitting was determined using the open circuit respiration chamber (RC). The time taken for similar activities was also determined in an outdoor enclosure (OD). The animals were fed kangkong (Ipomoea aquatica), sweet potato (Ipomoea batatas) and rabbit pellet ad libitum. Male mouse deer consumed more dry matter (DM), organic matter (OM) and gross energy (GE) than female. The time for each activity of male and female mouse deer kept in RC and OD was similar. The average time spent in RC and OD for both male and female, respectively, for sitting (956 and 896 min/day) was significantly (P<0.01) longer than standing (463 and 520 min/day) and eating (21 and 24 min/day). Heat production for male and female mouse deer, respectively, during eating was the highest (0.44 and 0.43 kJ/kg W^0.75/min) followed by standing (0.37 and 0.33 kJ/kgW^0.75/min) and sitting (0.26 and 0.26 kJ/kg W^0.75/min). The difference in HP per min during standing between male and female was significant (P<0.05). The HP for 08.00–14.00 h and 14.00–20.00 h periods were higher than 20.00–02.00 h and 02.00–08.00 h periods. The overall HP for males during 08.00–14.00 h and 14.00–20.00 h periods were significantly (P<0.05) higher (114.8 and 119.2 kJ/kg W^0.75) than female (107.5 and 110.4 kJ/kg W^0.75), respectively.     

Effect of gibberellic acid on carnation flower senescence: evidence that the delay of carnation flower senescence by gibberellic acid depends on the stage of flower development

Gibberellic acid at concentrations of 10^–5 M and 10^–4 M delayed the senescence of cut carnation flowers, when applied continuously via the stem, to flowers between the closed brush and fully open stages of development. Older flowers with reflexed petals were unresponsive. Treatment with paclobutrazol, an inhibitor of GA biosynthesis, prevented tight buds from opening fully, reduced the longevity of partially open flowers, but was ineffective when applied continuously to fully open flowers. Gibberellic acid-treated flowers did not show simultaneous petal inrolling, a known indicator of senescence, and the time to complete petal drying was extended. Gibberellic acid modified the climacteric ethylene rise in a manner consistent with the extension of longevity. These results provide evidence for a correlative role of gibberellins in flower development.Abbreviations GA₃ gibberellin A₃ - GLC gas liquid chromatography     

Feeding the fairy shrimp Streptocephalus (Anostraca - Crustacea) with the rotifer Anuraeopsis

Laboratory-cultured Streptocephalus torvicornis were offered 8 concentrations (from 6 to 800 ind. ml^–1) of Anuraeopsis fissa for periods of 2 h 30 min. Two size classes, small (male: 14.7 mm± 1.6, female: 15.4 mm± 1.3) and large (male: 20.0 mm±2.0, female: 23.1 mm± 1.5), of S. torvicornis were used. Functional response for large S. torvicornis (both sexes) plateaued at 400 rotifers ml^–1, while in small specimens it did so at 200 prey ml^–1. Females consumed significantly more (30%) prey than males. Large males consumed maximum 4730 rotifers h^–1, females 6560 h^–1.     

Oxygen consumption and ammonia accumulation in the hypolimnion of Walker Lake, Nevada

Walker Lake (area = 140 km², Z _mean = 19.3 m) is a large, terminal lake in western Nevada. As a result of anthropogenic desiccation, the lake has decreased in volume by 75% since the 1880s. The hypolimnion of the lake, now too small to meet the oxygen demand exerted by decaying matter, rapidly goes anoxic after thermal stratification. Field and laboratory studies were conducted to examine the feasibility of using oxygenation to avoid hypolimnetic anoxia and subsequent accumulation of ammonia in the hypolimnion, and to estimate the required DO capacity of an oxygenation system for the lake. The accumulation of inorganic nitrogen in water overlaying sediment was measured in laboratory chambers under various DO levels. Rates of ammonia accumulation ranged from 16.8 to 23.5 mg-N m^–2 d^–1 in chambers with 0, 2.5 and 4.8 mg L^–1 DO, and ammonia release was not significantly different between treatments. Beggiatoa sp. on the sediment surface of the moderately aerated chambers (2.5 and 4.8 mg L^–1 DO) indicated that oxygen penetration into sediment was minimal. In contrast, ammonia accumulation was reversed in chambers with 10 mg L^–1 DO, where oxygen penetration into sediment stimulated nitrification and denitrification. Ammonia accumulation in anoxic chambers (18.1 and 20.6 mg-N m^–2 d^–1) was similar to ammonia accumulation in the hypolimnion from July through September of 1998 (16.5 mg-N m^–2 d^–1). Areal hypolimnetic oxygen demand averaged 1.2 g O₂ m^–2 d^–1 for 1994–1996 and 1998. Sediment oxygen demand (SOD) determined in experimental chambers averaged approximately 0.14 g O₂ m^–2 d^–1. Continuous water currents at the sediment-water interface of 5–6 cm s^–1 resulted in a substantial increase in SOD (0.38 g O₂ m^–2 d^–1). The recommended oxygen delivery capacity of an oxygenation system, taking into account increased SOD due to mixing in the hypolimnion after system start-up, is 215 Mg d^–1. Experimental results suggest that the system should maintain high levels of DO at the sediment-water interface (10 mg L^–1) to insure adequate oxygen penetration into the sediments, and a subsequent inhibition of ammonia accumulation in the hypolimnion of the lake.     

Periphyton as a potential phosphorus sink in the Everglades Nutrient Removal Project

Phosphorus uptake and release by periphyton mats were quantified in the Everglades Nutrient Removal Project (ENRP) to evaluate the potential for periphyton P removal. Short-term P uptake rates were determined by incubating cyanobacteria (Oscillatoria princeps and Shizothrix calcicola) and Chlorophycean (primarily Rhizoclonium spp.) algal mat samples for 0.5–2 h under ambient conditions in BOD bottles spiked with soluble reactive P (SRP). Cyanobacterial mats removed P more than twice as fast (80–164 μg P h⁻¹ g⁻¹ AFDM) as Chlorophycean mats (33–61 μg P h⁻¹ g⁻¹ AFDM) during these incubations. In a longer term study, fiberglass cylinders were used to enclose 1.8 m² plots within the wetland and were dosed weekly for 7 weeks with: (1) no nutrients; (2) SRP (0.25 g P m⁻² week⁻¹); or (3) SRP plus nitrate (0.42 g N m⁻² week⁻¹) and ammonium (0.83 g N m⁻² week⁻¹). Phosphorus uptake rates by this periphyton assemblage, which was dominated by the chlorophytes Stigeoclonium spp. and Oedogonium spp., were measured weekly and were similar among nutrient treatments on most dates, indicating that the algal storage compartment for P was not saturated despite repeated P additions. Decomposition rates and P loss by cyanobacteria and Chlorophycean mats were determined by measuring biomass loss and SRP release in darkened BOD bottles over 28–42 day periods under anaerobic and aerobic conditions. First-order aerobic and anaerobic decomposition rates for cyanobacterial mats (k = 0.1095 and 0.1408 day⁻¹, respectively) were 4–20-fold higher than rates for Chlorophycean mats (k = 0.0066 and 0.0250 day⁻¹, respectively) and cyanobacteria released considerably more P back to the water column. Our findings suggest that periphyton can be an important short-term sink for P in treatment wetlands and that retention is strongly affected by the taxonomic composition of the periphyton assemblage.     

Effects of aqueous hop (Humulus Lupulus L.) extract on vascular reactivity in rats: Mechanisms and influence of gender and hormonal status

Phytoestrogens, naturally occurring plant compounds having oestrogenic and/or anti-oestrogenic activity, are present in many human foodstuffs including hop. Moderate intakes of isoflavonoid phytoestrogens have been associated with a reduction in cardiovascular diseases incidence. So, it is possible that hop (Humulus Lupulus L.) might similarly contribute to the reported health-beneficial effects of moderate beer consumption. Thus, the purpose of this study was to investigate in vitro effects of aqueous hop extract on thoracic vascular reactivity in Sprague Dawley male and female rats. Endothelium-intact thoracic arterial rings from male rats (MALE, n=8), sham-ovariectomized (Sham OVX) female (n=8) and ovariectomized (OVX) female rats (n=8) were used. We assessed the relaxation induced by aqueous hop extract (10⁻⁹, 10⁻² g/l) in aortic rings precontracted with norepinephrine (10⁻⁷ M), in the absence or in the presence of l-NAME (10⁻⁴ M), indomethacin (10⁻⁵ M), thapsigargin (10⁻⁴ M), iberiotoxin (3.10⁻⁸ M), apamin (3.10⁻⁸ M) and TEA (3.10⁻⁴ M). Aqueous hop extract induced relaxation of endothelium-intact thoracic arterial rings in MALE and Sham OVX rats, whereas a weak effect was observed in OVX rats. This vasorelaxation was strongly inhibited in presence of l-NAME, indomethacin and thapsigargin. These data indicated that aqueous hop extract-induced vasodilation, in male and intact female rats, is mediated by NOS activation, cyclooxygenase products and Ca²⁺ pathways. Moreover, our results suggested that effect of hop in enhancing vascular reactivity was independent of gender but strongly related to hormonal status.     

Benthic oxygen flux in the highly productive subarctic Lake Myvatn, Iceland: In situ benthic flux chamber study

In situ paired light and dark-stirred benthic flux chambers were used to estimate dissolved oxygen flux across the sediment–water interface in Lake Mývatn, Iceland. Three sampling stations were selected, each station reflecting a specific sedimentary environment, benthic communities, and water depth. During this study the phytoplankton density was low. Spatial and seasonal variations of bottom DO concentration and DO flux have been observed during this study. The oxygen consumption rate at all study sites had a mean of –89 (±44) mmol m^–2 d^–1 while the oxygen production rate due to benthic algae had a mean of 131 (±103) mmol m^–2 d^–1. There was a strong correlation (r=0.91) between oxygen consumption rate and temperature. This was presumably because of the temperature influence on rates of microbial and macrobenthic processes. The mean benthic primary production rate at all study sites was 1216 (±957) mg C m^–2 d^–1 between June 2000 and February 2001. Annual gross benthic primary production was estimated from the gross mean daily benthic DO production (P) and Redfield's C:O₂ ratio of 106:138 to be 420 g C m^–2 y^–1 at station HO, 250 g C m^–2 y^–1 at B2 and 340 g C m^–2 y^–1 at station 95. Thus, the mean gross benthic primary production was estimated as 1151 mg C m^–2 d^–1 at station HO, 685 mg C m^–2 d^–1 at station B2, and 932 mg C m^–2 d^–1 at station 95.     

Time-activity budgets and energetics of Dipper Cinclus cinclus are dictated by temporal variability of river flow

The white-throated Dipper (Cinclus cinclus) is unique among passerine birds by its reliance on diving to achieve energy gain in fast-flowing waters. Consequently, it should have evolved behavioural adaptations allowing responding directly to runoff patterns (one of the assumptions of the Natural Flow Regime Paradigm—NRFP). In this study (October 1998–August 2001), we investigated how behavioural and energy use strategies in Dippers might vary under the natural flow regime of snowmelt-dominated streams in The Pyrénées (France) where natural flow regime is highly seasonal and predictable. We recorded time spent in each of 5 behavioural activities of ringed birds to estimate time–activity budgets and derive time–energy budgets enabling the modelling of daily energy expenditure (DEE). Annual pattern in ‘foraging’ and ‘resting’ matched perfectly the annual pattern of the natural regime flow and there was a subtle relationship between water stage and time spent ‘diving’ the later increasing with rising discharge up to a point where it fell back. Thus, time–activity budgets meet the main prediction of the NRFP. For males and females Dippers, estimates of feeding rates (ratio E_obs/E_req = observed rate of energy gain / required foraging rate) and energy stress (M = DEE / Basal Metabolic Rate) also partly matched the NFRP. Maximum value for the ratio E_obs/E_req was registered in May whilst M peaked in spring. These ratios indicated that Pyrenean Dippers could face high energy stress during winter but paradoxically none during high snowmelt spates when food is expected to be difficult to obtain in the channel and when individual birds were observed spending ca 75% of the day ‘resting’. Annual pattern in DEE did not match the NFRP ; two phases were clearly identified, the first between January to June (with oscillating values 240–280 kJ d^− 1 ind^− 1) and the second between July and December (200–220 kJ d^− 1 ind^− 1). As total energy expenditure was higher during the most constraining season or life cycle, we suggest that energy management by Dippers in Pyrenean mountain streams may fit the ‘peak total demand’ hypothesis. At this step of the study, it is not possible to tell whether Dippers use an ‘energy-minimisation’ or an ‘energy-maximisation' strategy.     

Controlled Environment Chambers for Investigating Tree Response to Elevated CO2 and Temperature Under Boreal Conditions

A closed CO₂ and temperature-controlled, long-term chamber system has been developed and set up in a typical boreal forest of Scots pine (Pinus sylvestris L.) near the Mekrijärvi Research Station (62°47N, 30°58E, 145 m above sea level) belonging to the University of Joensuu, Finland. The main objectives of the experiment were to provide a means of assessing the medium to long-term effects of elevated atmospheric CO₂ concentration (EC) and temperature (ET) on photosynthesis, respiration, growth, and biomass at the whole-tree level and to measure instantaneous whole-system CO₂ exchange. The system consists of 16 chambers with individual facilities for controlling CO₂ concentration, temperature, and the combination of the two. The chambers can provide a wide variety of climatic conditions that are similar to natural regimes. In this experiment the target CO₂ concentration in the EC chambers was set at a fixed constant of 700 µmol mol^–1 and the target air temperature in the ET chambers to track the ambient temperature but with a specified addition. Chamber performance was assessed on the base of recordings covering three consecutive years. The CO₂ and temperature control in these closed chambers was in general accurate and reliable. CO₂ concentration in the EC chambers was within 600–725 µmol mol^–1 for 90 % of the exposure time during the "growing-season" (15 April – 15 September) and 625–725 µmol mol^–1 for 88 % of the time in the "off-season" (16 September – 14 April), while temperatures in the chambers were within ±2.0 °C of the ambient or target temperature in the "growing season" and within ±3.0 °C in the "off season". There were still some significant chamber effects. Solar radiation in the chambers was reduced by 50–60 % for 82 % of the time in the "growing season" and 55–65 % for 78 % of the time in the "off season", and the relative humidity of the air was increased by 5–10 % for 72 % of the time in the "growing season" and 2–12 % for 91 % of the time in the "off season". The crown architecture and main phenophase of the trees were not modified significantly by enclosure in the chambers, but some physiological parameters changed significantly, e.g., the radiant energy-saturated photosynthesis rate, transpiration rate, maximum photochemical efficiency of photosystem 2, and chlorophyll content.     

A novel photobioreactor with transparent rectangular chambers for cultivation of microalgae

An open tank photobioreactor containing transparent rectangular chambers (TRCs) was developed to improve the photosynthetic efficiency of microalgal cultivation. The TRCs, made of transparent acrylic, conducted light deep into the photobioreactor, especially at high cell concentrations. The average irradiance, I_av, was calculated by Lambert–Beer's law, and was used to determine the light conditions in the cultivation system. The photobioreactor provided large areas of illumination that improved the effective utilization of light energy for microalgae growth and created a good artificial environment for a high rate of cell growth, even at low I_av. The biomass concentration of Chlorella sp. reached 3.745 g L⁻¹ on the 13th day, with biomass productivity of 0.340 g L⁻¹ d⁻¹. The total biomass obtained was 56% more than that of similar culture systems without TRCs.     

Revegetation and reclamation of soils using wild leguminous shrubs in cold semiarid Mediterranean conditions: Litterfall and carbon and nitrogen returns under two aridity regimes

This study was designed to evaluate the litter produced by Mediterranean shrub legumes subjected to two conditions of aridity. Seasonal litterfall patterns and litter chemistry showed no significant variation with soil aridity. The effects of aridity on the amount of litter produced were related to the plant species. A higher availability of water led to a 110% increase in litter production by Colutea arborescens (3191 vs. 1516 kg ha^–1) and to a 24% increase for Medicago strasseri (5288 vs. 4258 kg ha^–1). The litter provided by Colutea cilicica failed to significantly increase (1651 vs. 1825 kg ha^–1) in less arid conditions. In our experimental conditions, Dorycnium hirsutum showed high mortality and scarce persistence. In general, the litter supplied by these shrub legumes was low in lignin and showed high levels of easily degradable organic-C. Its N content, in the range 18 to 26.5 g kg^–1, was similar to that described by others for multipurpose tropical legumes. Under the semi-arid conditions of central Spain, C. arborescens and C. cilicica gave rise to potential yearly returns of 662 and 693 kg ha^–1 organic-C, and 35 and 44 kg ha^–1 N, respectively. M. strasseri provided a yearly organic-C return (1742 kg ha^–1) similar to that of a mature Mediterranean wood, and to a potential N return (78 kg ha^–1) that substantially exceeded this reference. Compared to the sclerophyllous species typical of the Mediterranean environment, shrub legumes show a much greater potential for enhancing N and organic-C levels, and consequently, for improving the biological activity of degraded soils. This feature is thought to be associated with the rapid and constant renewal of their leaves and their ability to provide the soil with other easily degraded materials.     

Sex differences in the biotransformation of 2-acetylaminofluorene in cultured rat hepatocytes

Sex-related differences in susceptability to 2-acetylaminofluorene (2-AAF) hepatocarcinogenicity and in vivo biotransformation of 2-AAF have been observed. In order to determine the contribution of hepatocytes to these differences, 2-AAF biotransformation was investigated in monolayer cultures of hepatocytes freshly isolated from male and female F-344 rats.In cultured hepatocytes from both sexes, ring and N-hydroxylated, deacetylated and conjugated metabolites were formed. The half-life of 2-AAF was similar at concentrations of 5×10^–6 and 10^–5 M; however, at 10^–4 M a slower rate was observed in cultures from males. Although the total formation of aqueous metabolites was similar, the ratio of sulfate to glucuronide conjugates of 2-AAF formed by hepatocytes from male and female rats differed. Sulfate conjugates predominated in hepatocytes from male rats, whereas in females, glucuronides predominated. The demonstration of sex-dependent variations in the rate of metabolism at a high concentration of 2-AAF and in conjugation provides evidence that in vivo differences are a function, at least in part, of the biotransformation characteristics of hepatocytes.Abbreviations 2-AAF 2-acetylaminofluorene - 2AF 2-aminofluorene - WME Williams Medium E.     

The effects of ghrelin on the in vitro spontaneous and sGnRH-A stimulated luteinizing hormone (LH) release from the pituitary cells of common carp (Cyprinus carpio L.)

In fish, like in mammals, ghrelin affects gonadotropin release acting at the level of the hypothalamus as well as directly on the pituitary gland. In the present study, enzymatically dispersed pituitary cells obtained from sexually mature male and female carp (Cyprinus carpio L.) were incubated in the presence of human ghrelin at the concentration of 10^− 7 or 10^− 6 M, salmon GnRH analogue (Des-Gly¹⁰, D-Arg⁶, Trp⁷, Leu⁸, Pro⁹)-LHRH (sGnRH-A) at the concentration of 10^− 8 M or the combination of ghrelin (both concentrations) and sGnRH-A. ELISA method was used for carp LH levels determination in the media collected after 10 or 24 h of incubation. Ghrelin at the concentration of 10^− 6 M caused the increase of the spontaneous LH secretion from female pituitary cells only. The combination of ghrelin (both concentrations) with sGnRH-A resulted in the significant elevation of LH levels in the incubations of both male and female pituitary cells in comparison with control incubations as well as with sGnRH-A alone treated cells. The results obtained in this study show that ghrelin functions as LH-stimulating hormone in common carp and that it acts directly on gonadotrophic cells, potentiating also the action of GnRH.     

ICChI, a glycosylated chitinase from the latex of Ipomoea carnea

A multi-functional enzyme ICChI with chitinase/lysozyme/exochitinase activity from the latex of Ipomoea carnea subsp. fistulosa was purified to homogeneity using ammonium sulphate precipitation, hydrophobic interaction and size exclusion chromatography. The enzyme is glycosylated (14–15%), has a molecular mass of 34.94 kDa (MALDI–TOF) and an isoelectric point of pH 5.3. The enzyme is stable in pH range 5.0–9.0, 80 °C and the optimal activity is observed at pH 6.0 and 60 °C. Using p-nitrophenyl-N-acetyl-β-d-glucosaminide, the kinetic parameters K_m, V_max, K_cat and specificity constant of the enzyme were calculated as 0.5 mM, 2.5 × 10⁻⁸ mol min⁻¹ μg enzyme⁻¹, 29.0 s⁻¹ and 58.0 mM⁻¹ s⁻¹ respectively. The extinction coefficient was estimated as 20.56 M⁻¹ cm⁻¹. The protein contains eight tryptophan, 20 tyrosine and six cysteine residues forming three disulfide bridges. The polyclonal antibodies raised and immunodiffusion suggests that the antigenic determinants of ICChI are unique. The first fifteen N-terminal residues G–E–I–A–I–Y–W–G–Q–N–G–G–E–G–S exhibited considerable similarity to other known chitinases. Owing to these unique properties the reported enzyme would find applications in agricultural, pharmaceutical, biomedical and biotechnological fields.     

Biology, life table and predation of Feltiella acarisuga (Diptera: Cecidomyiidae) feeding on Tetranychus cinnabarinus eggs (Acari: Tetranychidae)

Feltiella acarisuga (Vallot) is a common predatory gall midge, which feeds on many species of spider mites. All major life history and life table parameters of F. acarisuga were determined using the carmine spider mite, Tetranychus cinnabarinus (Boisduval) eggs as prey under laboratory conditions [26.7 ± 2 °C, 75 ± 5% RH, and a photoperiod of l4:10 (L:D) h]. Developmental times of F. acarisuga were 2.6, 7.1, and 6.7 d for eggs, larvae, and pupae, respectively, with an average of 16.4 d from egg to adult emergence. Female immatures development took ≈1 d longer than male immatures did. Adult F. acarisuga lived an average of 12.8 d, and the female adults (13.3 d) lived significantly longer than male adults did (11.9 d). After an average of 1.5 d preoviposition period, each female laid an average of 27.3 eggs in its life span with an average of 2.1 eggs per day and an average of 2.8 eggs on each of its oviposition day. The raw data were analyzed using an age-stage, two-sex life table method that takes into consideration of the variable developmental rates among individuals and between sexes. The intrinsic rate of natural population increase (r), net reproductive rates (R_o), gross reproductive rate (∑m_x), generation time (T), and doubling time (DT), and the finite rate of increase (λ) of F. acarisuga were estimated using the age-stage, two-sex (male and female) life table analysis as 0.122 d⁻¹, 16.19 eggs per female, 20.81 eggs per female, 22.81 d, 1.1298 d, and 5.7 d⁻¹, respectively. Each of the first, second, and third instar larvae of F. acarisuga consumed an average of 35.5, 54.0 and 86.9 T. cinnabarinus eggs per day, respectively. Larvae of F. acarisuga could consume an average of 175.4, T. cinnabarinus eggs, and female larvae consumed 14% more spider mite eggs (187.6 eggs) than male larvae (165.1 eggs). The significance of other life table parameters related to the population and the potential of using F. acarisuga as a biological control agent are discussed.     

Effect of prey stage on life-table attributes of a genetically manipulated strain ofMetaseiulus occidentalis (Acari: Phytoseiidae)

Metaseiulus occidentalis females from the carbaryl-organophosphate-sulfur resistant strain (COS) lived longer (25.3 days versus 19.7 days), had a higher total fecundity (43.8 eggs female^–1 versus 33.6 eggs female^–1) and a higher daily fecundity rate (2.4 eggs female^–1 day^–1 versus 2.0 eggs female^–1 day^–1), and exhibited a higher intrinsic rate of increase (0.243 individuals female^–1 day^–1 versus 0.182 individuals female^–1 day^–1) and shorter generation time (13.9 days versus 17.0 days), at 24–28°C, 47–56%rh under continuous fluorescent light, when reared on a diet of 0–48-h-old eggs rather than a diet of mixed actives ofTetranychus pacificus McGregor on bean leaf disks. The sexratio of the progeny was female-biased for both diets, 2.1 females to 1 male forM. occidentalis reared on eggs and 2.0:1 : forM. occidentalis reared on mixed actives, suggesting that diet influences sex-ratio in some unknown way.There was no significant difference in oviposition rates for repeatedly-mated and once-matedM. occidentalis females reared on a diet of younger (0–24-h old) eggs compared to a diet of older (72–96-h old) eggs ofT. pacificus.The COS strain ofM. occidentalis exhibited life-table parameters comparable to the other strains reported in the literature, suggesting that the reproductive attributes of this acarine predator were not reduced as a result of artificial laboratory selection. Diet, a biotic factor, produced substantial differences in life-table parameters, suggesting that this factor can influence conclusions regarding the potential efficacy of biological control agents.     

Gene expression in Mercurialis annun flowers: In vitro translation and sex genotype specificity. Male-specific cDNA cloning and hormonal dependence of a corresponding specific RNA

Summary Sequences specifically expressed in flowers of each sex type of Mercurialis annua have been demonstrated by a comparative analysis of the translation products of their poly(A)⁺ RNA populations (wheat germ system, two-dimensional electrophoresis). This method confirms previous results of hybridization kinetics: the staminate flowers of the normal fertile male (wild type) and restored fertile male strain (identical morphology) and those of the sterile male genotype contain specific poly(A)⁺ RNAs and sequences shared by sets of two of these males, as well as numerous common sequences. The pistillate flowers of the constructed female strain 19_–5 (carrying male sterility determinants) also contains specific poly(A)⁺ RNA compared to identical flowers of a normal female genotype. In vitro translation, however, showed a specificity (not revealed by hybridization kinetics) in the normal female genotype compared to a normal male genotype and to the female strain 19_–2 (female strain cDNAs hybridized 100% to poly(A)⁺ RNAs of male and female 19_–5 genotypes). Cloning certain specific sequences (cDNAs) of the fertile male wild type and using one cDNA as probe also confirms the previously described male specificity. Moreover, the hormonal dependence of RNA corresponding to a specific male probe is demonstrated: its kinetics of disappearance are a function of the action of the feminizing hormone (cytokinins). These results are in agreement with our hypothesis of sexual organogenesis: sexual hormones controlled by regulator genes of the sexual genotype induce, in definite cell lineages of bipotential meristems, the expression of genes specific for male or female expression.     

Surface-enhanced Raman spectroscopy combined with atomic force microscopy for ultrasensitive detection of thrombin

Indole-3-acetic acid (IAA) amide conjugates play an important role in balancing levels of free IAA in plant cells. The GH3 family of proteins conjugates free IAA with various amino acids. For example, auxin levels modulate expression of the Oryza sativa (rice) GH3-8 protein, which acts to prevent IAA accumulation by coupling the hormone to aspartate. To examine the kinetic properties of the enzyme, we developed a liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay system. Bacterially expressed OsGH3-8 was purified to homogeneity and used to establish the assay system. Monitoring of the reaction confirms the reaction product as IAA–Asp and demonstrates that production of the conjugate increases proportionally with both time and enzyme amount. Steady-state kinetic analysis using the LC–MS/MS-based assay yields the following parameters: V/E_t^IAA = 20.3 min⁻¹, K_m^IAA = 123 μM, V/E_t^ATP = 14.1 min⁻¹, K_m^ATP = 50 μM, V/E_t^Asp = 28.8 min⁻¹, K_m^Asp = 1580 μM. This is the first assignment of kinetic values for any IAA–amido synthetase from plants. Compared with previously described LC- and thin-layer chromatography (TLC)-based assays, this LC–MS/MS method provides a robust and sensitive means for performing direct kinetic studies on a range of IAA-conjugating enzymes.     

Measuring pad–pad pinch strength in a non-human primate: Macaca fascicularis

The primary purpose of this study was to establish a methodology for determining and perhaps predicting (via regression analysis of anthropometric measures) Macaca fascicularis isometric pinch strength for a specific task. The larger purpose of this work was to properly scale a pinching task for the monkeys in order to study dose-response relationships in a non-human primate model for carpal tunnel syndrome. Three female and one male macaque (n = 4) of varying size and age were trained to perform a left-handed pad–pad pinch. The task required 60° of wrist flexion at a static pinching distance of 3 cm between the thumb and fingers. Subjects were trained for a period of 20-weeks. After that time, an analysis of performance gradients found that they had each reached a plateau in their force output. Pinch strength for the four animals ranged from 29.4 to 59.8 N. Regression analysis revealed that body mass (kg) and wrist circumference (cm) were both predictive of pinch strength, exhibiting adjusted R² values of 0.93 (p = 0.024) and 0.96 (p = 0.015), respectively. Thus, the results suggest that maximal pinch strength could be acceptably estimated in future subjects using either the wrist circumference or the body mass measures, as both were strong predictors of pad–pad pinch strength.