The gaits of marsupials and the evolution of diagonal-sequence walking in primates.

OBJECTIVES: Documenting the variety of quadrupedal walking gaits in a variety of marsupials (arboreal vs. terrestrial, with and without grasping hind feet), to aid in developing and refining a general theory of gait evolution in primates. MATERIALS AND METHODS: Video records of koalas, ringtail possums, tree kangaroos, sugar gliders, squirrel gliders, wombats, numbats, quolls, a thylacine, and an opossum walking on a variety of substrates were made and analyzed to derive duty factors and diagonalities for symmetrical walking gaits. The resulting distributions of data points were compared with published data and theories. RESULTS: Terrestrial marsupials' gaits overwhelmingly plot slightly below the theoretical "horse line" (Cartmill et al., Zoological Journal of the Linnean Society. 2002;136:401-420) typical of terrestrial mammals; arboreal marsupials' gaits overwhelmingly plot more decisively above it. Both distributions are roughly parallel to the horse line, but arboreal animals exhibit increased diagonality, so that their higher-speed walking gaits overlap with those of typical primates on the Hildebrand diagram of diagonality against duty factor. CONCLUSIONS: Quadrupeds avoid gaits lying exactly on the (theoretically optimum) horse line, to avoid fore/hind limb interference ("forging"). This can be accomplished by either a slight reduction in diagonality ("downshifting") or a more decisive increase ("upshifting"). Tree-dwellers adopt the second option to eliminate unilateral bipods of support from the gait cycle. The upshifted horse line represents an early phase in the evolution of primate-like diagonal-sequence gaits.

Computer-assisted assessment of colonic polyp histopathology using probe-based confocal laser endomicroscopy.

INTRODUCTION: Probe-based confocal laser endomicroscopy (pCLE) is a promising modality for classifying polyp histology in vivo, but decision making in real-time is hampered by high-magnification targeting and by the learning curve for image interpretation. The aim of this study is to test the feasibility of a system combining the use of a low-magnification, wider field-of-view pCLE probe and a computer-assisted diagnosis (CAD) algorithm that automatically classifies colonic polyps. METHODS: This feasibility study utilized images of polyps from 26 patients who underwent colonoscopy with pCLE. The pCLE images were reviewed offline by two expert and five junior endoscopists blinded to index histopathology. A subset of images was used to train classification software based on the consensus of two GI histopathologists. Images were processed to extract image features as inputs to a linear support vector machine classifier. We compared the CAD algorithm's prediction accuracy against the classification accuracy of the endoscopists. RESULTS: We utilized 96 neoplastic and 93 non-neoplastic confocal images from 27 neoplastic and 20 non-neoplastic polyps. The CAD algorithm had sensitivity of 95%, specificity of 94%, and accuracy of 94%. The expert endoscopists had sensitivities of 98% and 95%, specificities of 98% and 96%, and accuracies of 98% and 96%, while the junior endoscopists had, on average, a sensitivity of 60%, specificity of 85%, and accuracy of 73%. CONCLUSION: The CAD algorithm showed comparable performance to offline review by expert endoscopists and improved performance when compared to junior endoscopists and may be useful for assisting clinical decision making in real time.

Expression of acquired macrolide resistance genes in Haemophilus influenzae.

OBJECTIVES: To investigate the phenotypic effect of expression of selected acquired macrolide resistance genes (AMRGs) in non-typeable Haemophilus influenzae (NTHi). METHODS: The AMRGs erm(A), erm(B) and erm(C) were cloned into Escherichia coli JM109 using the shuttle vector pLS88; constructed plasmids extracted from suitable clones were used to transform H. influenzae Rd by electroporation. Erythromycin and azithromycin MICs for suitable transformants were determined by broth microdilution. AMRG expression was determined using quantitative PCR on transformant cDNA with locked nucleic acid dual-labelled hydrolysis probes. RESULTS: Expression of all AMRGs was observed in the transformants. Some variation in expression between the AMRGs was apparent, but expression of all genes was associated with a notable increase in erythromycin and azithromycin MICs compared with untransformed H. influenzae Rd. CONCLUSIONS: While the establishment of erm genes within WT populations of NTHi remains contentious, H. influenzae is capable of expression of erm. Expression may be associated with a subsequent decreased susceptibility to macrolides in isolates and future monitoring of these genes in NTHi isolates is warranted.

Acquired macrolide resistance genes in Haemophilus influenzae?

OBJECTIVES: The objective of this study was to determine the prevalence of specific acquired macrolide resistance genes previously reported as present in clinical isolates of Haemophilus influenzae. METHODS: A collection of 172 clinical respiratory isolates of H. influenzae, including 59 isolates from cystic fibrosis patients and 27 from non-cystic fibrosis bronchiectasis patients with significant prior macrolide use, was established. This collection was tested for azithromycin susceptibility using Etest and screened for the presence of erm(A), erm(B), erm(C), erm(F), mef(A) and mef(E) using locked nucleic acid dual-labelled hydrolysis probes. RESULTS: The azithromycin MICs ranged from 0.09 to >256 mg/L, with 2 (1.2%) isolates susceptible, 163 (94.8%) intermediate and 7 (4%) resistant according to EUCAST breakpoints (susceptible, ≤0.12 mg/L; resistant, >4 mg/L). None of the acquired macrolide resistance genes erm(A), erm(B), erm(C), erm(F), mef(A) or mef(E) was detected in any of the isolates. CONCLUSIONS: The specific acquired macrolide resistance genes are not widespread in H. influenzae and the high prevalence of these genes previously reported might be unique to the specific circumstances of that study.

Contributions of roots and rootstocks to sustainable, intensified crop production.

Sustainable intensification is seen as the main route for meeting the world's increasing demands for food and fibre. As demands mount for greater efficiency in the use of resources to achieve this goal, so the focus on roots and rootstocks and their role in acquiring water and nutrients, and overcoming pests and pathogens, is increasing. The purpose of this review is to explore some of the ways in which understanding root systems and their interactions with soils could contribute to the development of more sustainable systems of intensive production. Physical interactions with soil particles limit root growth if soils are dense, but root-soil contact is essential for optimal growth and uptake of water and nutrients. X-ray microtomography demonstrated that maize roots elongated more rapidly with increasing root-soil contact, as long as mechanical impedance was not limiting root elongation, while lupin was less sensitive to changes in root-soil contact. In addition to selecting for root architecture and rhizosphere properties, the growth of many plants in cultivated systems is profoundly affected by selection of an appropriate rootstock. Several mechanisms for scion control by rootstocks have been suggested, but the causal signals are still uncertain and may differ between crop species. Linkage map locations for quantitative trait loci for disease resistance and other traits of interest in rootstock breeding are becoming available. Designing root systems and rootstocks for specific environments is becoming a feasible target.

Linking ascorbic acid production in Ribes nigrum with fruit development and changes in sources and sinks.

BACKGROUND AND AIMS: Understanding the synthesis of ascorbic acid (l-AsA) in green tissues in model species has advanced considerably; here we focus on its production and accumulation in fruit. In particular, our aim is to understand the links between organs which may be sources of l-AsA (leaves) and those which accumulate it (fruits). The work presented here tests the idea that changes in leaf and fruit number influence the accumulation of l-AsA. The aim was to understand the importance of leaf tissue in the production of l-AsA and to determine how this might provide routes for the manipulation of fruit tissue l-AsA. METHODS: The experiments used Ribes nigrum (blackcurrant), predominantly in field experiments, where the source-sink relationship was manipulated to alter potential leaf l-AsA production and fruit growth and accumulation of l-AsA. These manipulations included reductions in reproductive capacity, by raceme removal, and the availability of assimilates by leaf removal and branch phloem girdling. Natural variation in fruit growth and fruit abscission is also described as this influences subsequent experimental design and the interpretation of l-AsA data. KEY RESULTS: Results show that fruit l-AsA concentration is conserved but total yield of l-AsA per plant is dependent on a number of innate factors many of which relate to raceme attributes. Leaf removal and phloem girdling reduced fruit weight, and a combination of both reduced fruit yields further. It appears that around 50 % of assimilates utilized for fruit growth came from apical leaves, while between 20 and 30 % came from raceme leaves, with the remainder from 'storage'. CONCLUSIONS: Despite being able to manipulate leaf area and therefore assimilate availability and stored carbohydrates, along with fruit yields, rarely were effects on fruit l-AsA concentration seen, indicating fruit l-AsA production in Ribes was not directly coupled to assimilate supply. There was no supporting evidence that l-AsA production occurred predominantly in green leaf tissue followed by its transfer to developing fruits. It is concluded that l-AsA production occurs predominantly in the fruit of Ribes nigrum.

Hydraulic conductivity and PAT determine hierarchical resource partitioning and ramet development along Fragaria stolons.

Co-ordination of metabolic and physiological activity between plant parts is key to the control of growth and development. Here the movement of resources and their allocation between mother plants and daughter ramets along Fragaria stolons was quantified with respect to hierarchy. Gradients of internodal ramet leaf water potential (ψ) and stolon and ramet hydraulic conductivities (L) were measured together with apparent stolon IAA movement via the polar auxin transport pathway (PAT). These processes are linked with measurements of stolon vascular development. The pattern of tissue differentiation and lignification in sequential stele sections of stolons demonstrated the rapid acquisition of the capacity for water transport, with transpiration potentially varying systematically with stolon lignification and the acropetal decline in stolon xylem ψ. Stolon and ramet L declined acropetally, with L across older ramets being significantly lower than that of the connecting stolons. The capacity for polar IAA transport increased with stolon age; this was due to increased transport intensity in older tissues. The partitioning of dry matter was strongly hierarchical with younger ramets smaller than older ramets, while foliar concentrations of N, P, and K were greater for the younger ramets. The results show that stolon anatomy develops rapidly at the apical end, facilitating hierarchical ramet development, which is evident as a basipetal increase in L. The rapid development of transport tissue functionality enables young unrooted ramets to acquire water, in order to supply an expanding leaf area, as well as mineral ions disproportionally with respect to older ramets. This facilitates colonization and self-rooting of apical ramets. The unidirectional increase in basipetal PAT along stolons facilitates hierarchical ramet development.

Recombinant monoclonal antibody yield in transgenic tobacco plants is affected by the wounding response via an ethylene dependent mechanism.

Variability in recombinant IgG yield in transgenic tobacco plants has previously been observed in relation to leaf position, and is interpreted as a function of ageing and the senescence process, leading to increasing protein degradation. Here, similar findings are demonstrated in plants of different ages, expressing IgG but not IgG-HDEL, an antibody form that accumulates within the endoplasmic reticulum. Antibody yields declined following wounding in young transgenic plants expressing IgG but not in those expressing IgG-HDEL. However, in mature IgG plants, the opposite was demonstrated, with significant boosts in yield, while mature IgG-HDEL plants could not be boosted. The lack of response in IgG-HDEL plants suggests that the changes induced by wounding occur post-translationally, and the findings might be explained by wounding responses that differ in plants according to their developmental stages. Plant mechanisms involved in senescence and wounding overlap to a significant degree and compounds such as ethylene, jasmonic acid and salicylic acid are important for mediating downstream effects. Treatment of transgenic plants with ethylene also resulted in a decrease in recombinant IgG yield, which was consistent with the finding that wounded plants could induce lower IgG yields in neighbouring non-wounded plants. Treatment with 1-MCP, an ethylene antagonist, abrogated the IgG yield drop that resulted from wounding, but had no effect on the more gradual IgG yield loss associated with increasing plant age.

Optimisation of contained Nicotiana tabacum cultivation for the production of recombinant protein pharmaceuticals.

Nicotiana tabacum is emerging as a crop of choice for production of recombinant protein pharmaceuticals. Although there is significant commercial expertise in tobacco farming, different cultivation practices are likely to be needed when the objective is to optimise protein expression, yield and extraction, rather than the traditional focus on biomass and alkaloid production. Moreover, pharmaceutical transgenic tobacco plants are likely to be grown initially within a controlled environment, the parameters for which have yet to be established. Here, the growth characteristics and functional recombinant protein yields for two separate transgenic tobacco plant lines were investigated. The impacts of temperature, day-length, compost nitrogen content, radiation and plant density were examined. Temperature was the only environmental variable to affect IgG concentration in the plants, with higher yields observed in plants grown at lower temperature. In contrast, temperature, supplementary radiation and plant density all affected the total soluble protein yield in the same plants. Transgenic plants expressing a second recombinant protein (cyanovirin-N) responded differently to IgG transgenic plants to elevated temperature, with an increase in cyanovirin-N concentration, although the effect of the environmental variables on total soluble protein yields was the same as the IgG plants. Planting density and radiation levels were important factors affecting variability of the two recombinant protein yields in transgenic plants. Phenotypic differences were observed between the two transgenic plant lines and non-transformed N. tabacum, but the effect of different growing conditions was consistent between the three lines. Temperature, day length, radiation intensity and planting density all had a significant impact on biomass production. Taken together, the data suggest that recombinant protein yield is not affected substantially by environmental factors other than growth temperature. Overall productivity is therefore correlated to biomass production, although other factors such as purification burden, extractability protein stability and quality also need to be considered in the optimal design of cultivation conditions.

Root signals and stomatal closure in relation to photosynthesis, chlorophyll a fluorescence and adventitious rooting of flooded tomato plants.

BACKGROUND AND AIMS: An investigation was carried out to determine whether stomatal closure in flooded tomato plants (Solanum lycopersicum) results from decreased leaf water potentials (psi(L)), decreased photosynthetic capacity and attendant increases in internal CO(2) (C(i)) or from losses of root function such as cytokinin and gibberellin export. METHODS: Pot-grown plants were flooded when 1 month old. Leaf conductance was measured by diffusion porometry, the efficiency of photosystem II (PSII) was estimated by fluorimetry, and infrared gas analysis was used to determine C(i) and related parameters. KEY RESULTS: Flooding starting in the morning closed the stomata and increased psi(L) after a short-lived depression of psi(L). The pattern of closure remained unchanged when psi(;L) depression was avoided by starting flooding at the end rather than at the start of the photoperiod. Raising external CO(2) concentrations by 100 micromol mol(-1) also closed stomata rapidly. Five chlorophyll fluorescence parameters [F(q)'/F(m)', F(q)'/F(v)', F(v)'/F(m)', non-photochemical quenching (NPQ) and F(v)/F(m)] were affected by flooding within 12-36 h and changes were linked to decreased C(i). Closing stomata by applying abscisic acid or increasing external CO(2) substantially reproduced the effects of flooding on chlorophyll fluorescence. The presence of well-aerated adventitious roots partially inhibited stomatal closure of flooded plants. Allowing adventitious roots to form on plants flooded for >3 d promoted some stomatal re-opening. This effect of adventitious roots was not reproduced by foliar applications of benzyl adenine and gibberellic acid. CONCLUSIONS: Stomata of flooded plants did not close in response to short-lived decreases in psi(L) or to increased C(i) resulting from impaired PSII photochemistry. Instead, stomatal closure depressed C(i) and this in turn largely explained subsequent changes in chlorophyll fluorescence parameters. Stomatal opening was promoted by the presence of well-aerated adventitious roots, implying that loss of function of root signalling contributes to closing of stomata during flooding. The possibility that this involves inhibition of cytokinin or gibberellin export was not well supported.

Enhancement of artemisinin concentration and yield in response to optimization of nitrogen and potassium supply to Artemisia annua.

BACKGROUND AND AIMS: The resurgence of malaria, particularly in the developing world, is considerable and exacerbated by the development of single-gene multi-drug resistances to chemicals such as chloroquinone. Drug therapies, as recommended by the World Health Organization, now include the use of antimalarial compounds derived from Artemisia annua--in particular, the use of artemisinin-based ingredients. Despite our limited knowledge of its mode of action or biosynthesis there is a need to secure a supply and enhance yields of artemisinin. The present study aims to determine how plant biomass can be enhanced while maximizing artemisinin concentration by understanding the plant's nutritional requirements for nitrogen and potassium. METHODS: Experiments were carried out, the first with differing concentrations of nitrogen, at 6, 31, 56, 106, 206 or 306 mg L(-1) being applied, while the other differing in potassium concentration (51, 153 or 301 mg L(-1)). Nutrients were supplied in irrigation water to plants in pots and after a growth period biomass production and leaf artemisinin concentration were measured. These data were used to determine optimal nutrient requirements for artemisinin yield. KEY RESULTS: Nitrogen nutrition enhanced plant nitrogen concentration and biomass production successively up to 106 mg N L(-1) for biomass and 206 mg N L(-1) for leaf nitrogen; further increases in nitrogen had no influence. Artemisinin concentration in dried leaf material, measured by HPLC mass spectroscopy, was maximal at a nitrogen application of 106 mg L(-1), but declined at higher concentrations. Increasing potassium application from 51 to 153 mg L(-1) increased total plant biomass, but not at higher applications. Potassium application enhanced leaf potassium concentration, but there was no effect on leaf artemisinin concentration or leaf artemisinin yield. CONCLUSIONS: Artemisinin concentration declined beyond an optimal point with increasing plant nitrogen concentration. Maximization of artemisinin yield (amount per plant) requires optimization of plant biomass via control of nitrogen nutrition.

Cross-Sectional Imaging Techniques and Normal Anatomy of the Salivary Glands.

The salivary glands play an important role in digestion and oral hygiene, and give rise to a variety of benign and malignant pathologies. In suspected pathology, the goal of imaging is to confirm a lesion as being of salivary gland origin, narrow the list of differential considerations, define the extent of disease, and guide further management decisions. This review outlines the function, embryologic development, anatomy, and normal imaging features of the major salivary glands. The article also discusses imaging indications, the general approach to imaging the salivary glands, and the commonly used cross-sectional techniques used for evaluating the salivary gland.

Responses of seminal wheat seedling roots to soil water deficits.

The aims of this paper are to develop our understanding of the ways by which soil water deficits influence early wheat root growth responses, particularly how seminal roots respond to soil drying and the extent to which information on differences in soil water content are conveyed to the shoot and their impact on shoot behaviour. To achieve this, wheat seedlings have been grown, individually for around 25 days after germination in segmented soil columns within vertical plastic compartments. Roots were exposed to different soil volumetric moisture contents (SVMC) within the two compartments. Experiments where the soil in the lower compartment was allowed to dry to different extents, while the upper was maintained close to field capacity, showed that wheat seedlings allocated proportionally more root dry matter to the lower drier soil compartment. The total production of root, irrespective of the upper or lower SVMC, was similar and there were no detected effects on leaf growth rate or gas exchange. The response of seminal roots to proportionally increase their allocation of dry matter, to the drier soil was unexpected with such plasticity of roots system development traditionally linked to heterogeneous nutrient distribution than accessing soil water. In experiments where the upper soil compartment was allowed to dry, root growth slowed and leaf growth and gas exchange declined. Subsequent experiments used root growth rates to determine when seminal root tips first came into contact with drying soil, with the intentions of determining how the observed root growth rates were maintained as an explanation for the observed changes in root allocation. Measurements of seminal root ABA and ethylene from roots within the drying soil are interpreted with respect to what is known about the physiological control of root growth in drying soil.

Biochar Remediation Improves the Leaf Mineral Composition of Telfairia occidentalis Grown on Gas Flared Soil.

This study evaluates the effects of remediation of gas flared soil by biochar on the nutritional composition of cultivated Telfairia occidentalis leaves, relative to non-gas flared soil. Gas flared soils are degraded due to the presence of heavy metals, noxious gases, carbon soot and acidic rain. Biochar produced from oil palm fibre was applied at five different amounts: 0 t ha-1, 7.1 t ha-1, 13.9 t ha-1, 20.9 t ha-1 and 28.0 t ha-1 to containerized soils (both gas flared and control soil), inside a greenhouse, which were allowed to mineralize for two weeks. Two viable seeds of T. occidentalis per replicate were sown. After eight weeks of growth, leaves were harvested, dried and chemically analyzed. Application of biochar significantly increased leaf ash and crude fibre content of Telfairia occidentalis. Plants from soil treated with 13.9 t ha-1 of biochar had the highest concentrations of vitamins A, B1, B2, B6, C and E irrespective of soil type. Maximum increase in leaf vitamin and mineral content was obtained from leaves cultivated on gas flared soil treated with 13.9 t ha-1 and 7.1 t ha-1 of biochar respectively. The results show that biochar treatment can increase leaf mineral concentrations and that this effect is dependent on the amount of biochar application.

Sphenostylis stenocarpa (ex. A. Rich.) Harms., a Fading Genetic Resource in a Changing Climate: Prerequisite for Conservation and Sustainability.

The southeastern part of Nigeria is one of the major hotspots of useful plant genetic resources. These endemic species are associated with a rich indigenous knowledge and cultural diversity in relation to their use and conservation. Sphenostylis stenocarpa (ex. A. Rich.) Harms., (African Yam Bean (AYB)), is one such crop within the family of Fabaceae. Its nutritional and eco-friendly characteristics have value in ameliorating malnutrition, hidden hunger and environmental degradation inherent in resource-poor rural and semi-rural communities throughout Africa. However, lack of information from the custodians of this crop is limiting its sustainable development. Therefore, ethnobotanical surveys on the diversity, uses, and constraints limiting the cultivation and use of the crop in southeastern Nigeria were carried out. Five-hundred respondents were randomly selected and data collected through oral interviews and focused group discussion (FGD). Semi-structured questionnaires (SSQ) were also used to elicit information from a spectrum of AYB users comprising community leaders, farmers, market women and consumers in five States. Results showed that the majority of the respondents lacked formal education and were of the age group of 40-50 years, while the female gender dominated with limited access to land and extension officers. Seed coat colour largely determined utilization. Long cooking time, requirement for staking materials, aging of farmers and low market demand were among the major constraints limiting further cultivation and utilization of AYB. In-situ conservation was by hanging dried fruits by the fireside, beside the house, storing in earthenware, calabash gourds, cans and bottles. It is concluded that there is urgent need to scale up conservation through robust linkages between contemporary scientific domains and indigenous peoples in order to harness and incorporate the rich indigenous knowledge in local communities for enhanced scientific knowledge, biodiversity conservation and its sustainable utilization for food security.

Mild asymptomatic intrahepatic biliary dilation after cholecystectomy, a common incidental variant.

OBJECTIVE: The purpose of this study is to evaluate the prevalence of intra- and extrahepatic ductal dilatation in asymptomatic individuals after cholecystectomy. METHODS AND MATERIALS: This IRB-approved retrospective cohort study evaluated the prevalence of intra- and extrahepatic biliary dilation in 77 consecutive post cholecystectomy patients who had CT obtained in the portal venous phase. These were then compared to age and sex matched control patients. Two radiologists in consensus blinded to surgical history evaluated the intrahepatic ducts qualitatively for dilatation. A single radiologist using the best of three orthogonal planes measured the extrahepatic ducts. Extrahepatic ducts were considered dilated if >7 mm plus 1 mm/decade after 60 years. T tests and chi-squared tests were performed. RESULTS: Cholecystectomy patient duct patterns: normal ducts 26% (20/77); intra- and extrahepatic dilation 31.2% (24/77); intrahepatic dilation only 18.2% (14/77); extrahepatic dilation only 24.7% (19/77). Control patient duct patterns: normal ducts 88.3% (68/77); intra- and extrahepatic dilation 2.6% (2/77); intrahepatic dilation only 2.6% (2/77); extrahepatic dilation only 6.5% (5/77). All intrahepatic ductal dilatation was mild. Total intrahepatic dilation: 49.4% (cholecystectomy); 5.2% (control patients). The relative risk of intrahepatic ductal dilation in cholecystectomy patients was 9.5:1. Increased prevalence of intra- and extrahepatic dilation in cholecystectomy patients was statistically significant (p < 0.0001). Average extrahepatic duct was 7.8 mm (cholecystectomy) and 5.3 mm (control patients) (p < 0.001). CONCLUSION: Mild intrahepatic biliary dilation in the setting of cholecystectomy is very common, and if not associated with clinical or biochemical evidence of obstruction is likely of no clinical significance.