Identification and Characterization of Cancer Stem Cells from Head and Neck Squamous Cell Carcinoma Cell Lines

Background/Aims: Head and neck squamous cell carcinoma (HNSCC) ranks sixth worldwide for tumor-related mortality. A subpopulation of tumor cells, termed cancer stem cells (CSCs), has the ability to support cancer growth. Therefore, profiling CSC-enriched populations could be a reliable tool to study cancer biology. Methods: We performed phenotypic characterization of 7 HNSCC cell lines and evaluated the presence of CSCs. CSCs from Hep-2 cell line and HNSCC primary cultures were enriched through sphere formation and sphere-forming cells have been characterized both in vitro and in vivo. In addition, we investigated the expression levels of Nicotinamide N-methyltransferase (NNMT), an enzyme overexpressed in several malignancies. Results: CSC markers were markedly expressed in Hep-2 cell line, which was found to be highly tumorigenic. CSC-enriched populations displayed increased expression of CSC markers and a strong capability to form tumors in vivo. We also found an overexpression of CSC markers in tumor formed by CSC-enriched populations. Interestingly, NNMT levels were significantly higher in CSC-enriched populations compared with parental cells. Conclusion: Our study provides an useful procedure for CSC identification and enrichment in HNSCC. Moreover, results obtained seem to suggest that CSCs may represent a promising target for an anticancer therapy

The golden native drone fly (Eristalinus punctulatus) is an effective hybrid carrot pollinator that lives within Australian crop agroecosystems

1. Native insect flower visitors can be important contributors to crop pollination, yet little is known of their pollination abilities and the resources (habitat) they need to be supported within crop agroecosystems. 2. Here, we compared the abundance and pollination abilities of the golden drone fly (Eristalinus punctulatus) to the European honey bee (Apis mellifera) in hybrid carrot crop fields known to produce variable seed yields in regional New South Wales, Australia. We further observed the egg-laying behaviours of female golden drone flies at a commercial berry orchard to provide insight into the habitat needs of this species. 3. In hybrid carrot crop fields, golden drone flies were far less abundant flower visitors than European honey bees" however, these flies deposited more carrot pollen grains on average (8.21±3.04SE) onto carrot flowers than European honey bees (3.45±1.06SE). Both insects also deposited pollen onto a similar number of carrot flowers (pollinated) per visit (about 2 out of 18). 4. Golden drone flies were observed laying eggs within masses of discarded red raspberry plant roots and soil (root balls) at a commercial berry orchard. The natural habitat utilised by these flies, as well as their egg-laying behaviours, were described for the first time. 5. Our results indicate that golden drone flies are effective pollinators of hybrid carrot crop plants. The habitat that these flies utilised to lay eggs (discarded plants and water) is cheap and commonly found in crop agroecosystems. Therefore, we recommend placing this low-cost habitat within, or nearby, crop fields as a potential management practice to support the lifecycle needs of golden drone flies and other non-bee pollinators

Bees visit less in the middle of polytunnels

Raspberry is a pollinator-dependent crop that relies on the transfer of pollen from insects to maximise fruit production and quality

The behaviour and movement of insect pollinators visiting blueberry plants

As most blueberry varieties benefit from Insect polllnation to provide higher yields and better fruit quality, it is important to understand how differences in insect communities and their behaviours in different regions influence the yield and quality of berries produced. Jeremy's honours project focused on pollinator communities in Mareeba, Far North Queensland (FNQ), to determine which were the main pollinators and how they respond to different tunnel types

Bee visitation and fruit quality in berries under protected cropping vary along the length of polytunnels

Wild and managed bees provide effective crop pollination services worldwide. Protected cropping conditions are thought to alter the ambient environmental conditions in which pollinators forage for flowers, yet few studies have compared conditions at the edges and center of growing tunnels. We measured environmental variables (temperature, relative humidity, wind speed, white light, and UV light) and surveyed activity of the managed honey bee, Apis mellifera L.; wild stingless bee, Tetragonula carbonaria Smith; and wild sweat bee, Homalictus urbanus Smith, along the length of 32 multiple open-ended polyethylene growing tunnels. These were spaced across 12 blocks at two commercial berry farms, in Coffs Harbour, New South Wales and Walkamin, North Queensland, Australia. Berry yield, fresh weight, and other quality metrics were recorded at discrete increments along the length of the tunnels. We found a higher abundance and greater number of flower visits by stingless bees and honey bees at the end of tunnels, and less frequent visits to flowers toward the middle of tunnels. The center of tunnels experienced higher temperatures and reduced wind speed. In raspberry, fruit shape was improved with greater pollinator abundance and was susceptible to higher temperatures. In blueberry, per plant yield and mean berry weight were positively associated with pollinator abundance and were lower at the center of tunnels than at the edge. Fruit quality (crumbliness) in raspberries was improved with a greater number of visits by sweat bees, who were not as susceptible to climatic conditions within tunnels. Understanding bee foraging behavior and changes to yield under protected cropping conditions is critical to inform the appropriate design of polytunnels, aid pollinator management within them, and increase economic gains in commercial berry crops

Observations of nectarivorous birds and potential biological control agents in berry orchards

Abstract We observed the brown honeyeater, Lichmera indistincta, visiting blueberry orchards in eastern Australia. L. indistincta is considered to be a nectarivorous bird, feeding almost entirely on nectar. However, we report observations of this species as both a nectarivore and insectivore in blueberry crops and as an insectivore in raspberry crops. Brown honeyeaters may be acting as biological control agents when feeding on leaf‐roller caterpillars (Lepidoptera: Tortricidae) in blueberry orchards and on brown blowflies (Calliphora stygia) in raspberry orchards. Although most blueberry growing regions use managed honey bees as the main pollinator, more studies on the complex tritrophic interactions occurring in these crops are needed to better understand the costs and benefits of different farm management practices upon alternative pollinators

The brown honeyeater (Lichmera indistincta) is an effective pollinator in Australian blueberry orchards

Abstract The brown honeyeater (Lichmera indistincta) is a common bird species in Australia and is a pollinator of many native plant species but there are no records demonstrating evidence of effective pollination by honeyeaters in commercial cropping systems. We observed L. indistincta visiting rabbiteye blueberry flowers at a commercial berry orchard on the Mid North Coast of New South Wales and evaluated their capacity to transfer pollen. We did this by bagging flowers to prevent visitation prior to anthesis; after anthesis, we unbagged the flowers and waited for L. indistincta visits. After a single visit, we collected the stigma and mounted it on a slide to evaluate stigmatic pollen load. We observed the birds visiting blueberry flowers and foraging on nectar. We also found that L. indistincta deposited similar amounts of pollen to other common insect pollinator species, such as honeybees and stingless bees. As the commercial berry orchard blocks are surrounded by diverse remnant vegetation, we hypothesize that the vegetation is likely providing habitat for these birds. Our results indicate that berry growers with management practices that protect and support remnant vegetation surrounding blueberry orchard blocks may benefit from higher blueberry pollination services by supporting diverse pollinators within fields

Protective nets reduce pollen flow in blueberry orchards

Protective covers are commonly employed in agricultural systems to reduce the impacts of extreme weather events, pest species and to control the environmental conditions in which crop plants are grown. As protected cropping systems are expanding rapidly, there is an urgent need to better understand how variations in netting practices might impact pollination service delivery by wild and managed insects to pollinator dependent crops. We used southern highbush blueberry (Vaccinium corymbosum L. interspecific hybrid) crops to investigate (i) how variations in protected cropping structures (fully netted, partially netted and unnetted blocks) influence the amount and composition of pollen deposited on crop stigmas; (ii) to what extent blueberry floral abundance and plant richness in remnant vegetation influence pollen composition on crop stigmas; and (iii) the difference between stigmatic pollen load composition in the middle and at the edge of crop blocks. We collected data from 15 field blocks of 6 different cultivars distributed on 10 farms. We collected blueberry stigmas to analyse the pollen load and measured blueberry floral abundance and richness of flowering plant taxa in remnant vegetation every two weeks. Our results indicate that blueberry pollen abundance on stigmas was reduced by up to 81% under full netting and 36% by partial netting. On blueberry stigmas, we identified a total of 31 morphospecies of non-blueberry pollen from 20 plant families. There was no relationship between blueberry stigmatic pollen loads and blueberry floral abundance. Moreover, the composition of non-blueberry pollen on stigmas differed between blueberry blocks under different netting categories. However, there was no relationship between plant taxa present in the surrounding remnant vegetation of each block and the pollen load on the stigmas of each block. Combining all netting treatments, stigmas located at the edge of the blocks received a greater amount of both conspecific (5% more) and heterospecific (40% more) pollen grains than those within the middle of blocks. Pollen flow in fields is reduced under netting structures as well as in the middle of blocks. Reduced blueberry pollen flow under nets may be detrimental to fruit yield and quality for some varieties of pollinator dependent crops, particularly those that are self-incompatible

Induced pluripotent stem cell-derived human macrophages as an infection model for Leishmania donovani.

The parasite Leishmania donovani is one of the species causing visceral leishmaniasis in humans, a deadly infection claiming up to 40,000 lives each year. The current drugs for leishmaniasis treatment have severe drawbacks and there is an urgent need to find new anti-leishmanial compounds. However, the search for drug candidates is complicated by the intracellular lifestyle of Leishmania. Here, we investigate the use of human induced pluripotent stem cell (iPS)-derived macrophages (iMACs) as host cells for L. donovani. iMACs obtained through embryoid body differentiation were infected with L. donovani promastigotes, and high-content imaging techniques were used to optimize the iMACs seeding density and multiplicity of infection, allowing us to reach infection rates up to 70% five days after infection. IC50 values obtained for miltefosine and amphotericin B using the infected iMACs or mouse peritoneal macrophages as host cells were comparable and in agreement with the literature, showing the potential of iMACs as an infection model for drug screening