Untreated Dishes Research Articles

Mitochondrial dysfunction significantly contributes to the sensitivity of tumor cells to anoikis and their metastatic potential

It is well-known that the survival of metastatic cells during their dissemination plays an important role in metastasis. However, does this mean that the final result of the metastatic cascade (the volume of metastatic damage to distant organs and tissues) depends on, or at least correlates with, the degree of resistance to anoikis (distinctive hallmarks of metastatic cells)? This question remains open.The aim of the work was to study in vitro the changes in the survival rates, proliferative activity, oxidative stress, and glycolysis intensity during three days of anchorage-dependent and anchorage-independent growth of two Lewis lung carcinoma cell lines (LLC and LLC/R9) and compare these changes with the status of mitochondria and metastatic potential of the cells in vivo. MethodsThe number and volume of lung metastases were estimated for each cell line after intramuscular inoculation of the cells in C57Bl/6 mice. For the in vitro study, the cells were seeded on Petri dishes pretreated with poly-HEMA or untreated dishes and then allowed to grow for 3 days. Cell viability, cell cycle progression, the level of reactive oxygen species (ROS), glucose consumption and lactate production rates were investigated daily in both growth conditions. An electron microscopy study of intracellular structures was carried out. ResultsThe study showed (as far as we know for the first time) a correlation between the metastatic potential of cells (determined in vivo) and their sensitivity to anoikis (assessed in vitro). The transition of LLC/R9 cells with an inherently defective mitochondrial system to the conditions of anchorage-independent growth was characterized by a decrease in survival, a slowdown in growth rates, an increase in both glucose consumption rate and intracellular ROS levels and manyfold lower metastatic potential, compared to highly metastatic LLC cells with the normal mitochondrial system.

Efficacy of Nets Coated with Different Concentrations of Alpha-Cypermethrin against Two Major Pests of Stored Tobacco

In the present study, we examined the insecticidal effect of Carifend® (BASF AG, Ludwigshafen, Germany; 163.2 mg m−2), an alpha-cypermethrin-coated polyester net, as well as Carifend-like nets containing different rates of alpha-cypermethrin, specifically 10, 30, 80, and 325 mg m−2, against two major stored tobacco insect pests, i.e., Lasioderma serricorne (Fabricius) (Coleoptera: Anobiidae) and Ephestia elutella (Hübner) (Lepidoptera: Pyralidae). The nets were glued at the bottom of plastic Petri dishes, and adults of L. serricorne and E. elutella were exposed to the nets for different exposure times (e.g., 1, 5, 15, 30, 60, 90, and 120 min, as well as 12 and 24 h). After each exposure interval, insect mortality was recorded, whereas after the final evaluation, alive individuals were transferred to untreated dishes, and delayed mortality was recorded after an additional period of 1, 3, 5, and 7 d. Based on our results, efficient control of both insect species was achieved with all nets, even the one containing a lower concentration of alpha-cypermethrin (10 mg m−2). High mortality rates were recorded for all treatments, indicating that even short exposure of adults of both tested species to the alpha-cypermethrin-coated nets tested can lead to mortality. For instance, at 30 mg m−2, mortality of E. elutella reached 40% 1 d after exposure for 30 min. Moreover, at the highest concentration (325 mg m−2), mortality reached 70% and 99% 7 d post exposure for 24 h of L. serricorne and E. elutella, respectively. Based on these data, Carifend®, as well as the rest of the Carifend-like nets tested, can be used as an alternative insecticide method for the control of two major stored tobacco insect species, i.e., L. serricorne and E. elutella. Insecticide treated nets can be used in areas in which fumigants and contact insecticides are not accessible.

Delayed responses of major stored-product beetle species after exposure to an alpha-cypermethrin-coated net

Carifend® is an alpha-cypermethrin-coated net that has been evaluated with success for the control of insects that infest stored tobacco. More recent data indicate that nets that contain insecticides can be further utilized against a wide range of stored-product insect species in different application scenarios. The aim of the current study was to evaluate Carifend® against adults of the rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), the granary weevil, Sitophilus granarius (L.) (Coleoptera: Curculionidae), the confused flour beetle, Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae), the saw-toothed grain beetle, Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae), the larger grain borer, Prostephanus truncatus (Horn) (Coleoptera: Bostrychidae), and the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae). Adults of these species were exposed to dishes containing Carifend® for 2, 8 and 24 h, and then they were transferred to untreated dishes, where mortality was recorded 7 d later. In most of the cases, the duration of the initial exposure increased mortality, but, in general, mortality was further increased after the termination of the post-exposure interval. Among the species tested, T. confusum was found to be the least susceptible to Carifend®. For instance, the mortality level was 5.5% for T. confusum even 7 d after exposure, while for P. truncatus it was 100%. The results of the present study clearly demonstrate that Carifend® can be an effective tool against a wide range of major stored-product beetle species, even at exposures that last for <24 h.

Ecophysiology of Alternaria alternata Causing Diseases in Litchi (Litchi chinensis)

Alternaria alternata (Fr.) Keissl. is an important pathogen of litchi (Litchi chinensis Sonn.) that causes blight of leaf, panicle and fruits. Ecophysiological determinants favouring growth and sporulation are keys to understand the pathogen behaviour. Therefore, studies were conducted to assess the effect of different medium, carbon source, pH, temperature, relative humidity and exposure to UV radiation on growth and sporulation of A. alternata under in vitro conditions. Results showed that the maximum mycelia growth rate was on Host Leaf Extract Dextrose Agar but sporulation was the maximum on Potato Dextrose Agar medium. The mycelia growth was more with dextrose as carbon source but the maximum dry weight was recorded in maltose amended medium. Optimum pH for mycelia growth was 6.0, and below or above this pH, mycelia growth was slow. It was evident that the maximum mycelia growth rate and germination of conidia were at 30 °C. The thermal death point of A. alternata was found to be 45 °C with 10 min. exposure. The maximum mean growth rate and germination percent was recorded at 100% relative humidity. The growth rate decreased with increasing exposure time to UV‑B rays up to 20 min, but at exposure time 25 and 30 min, colony changed to profuse mycelia growth with smaller sporulating zone in the centre compared to untreated dishes. The findings of the study will be helpful in understanding epidemiology and devising management strategies of this disease.

Aerosol concentration, deposition, particle size, and exposure interval as mortality factors Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae)

A series of trials was conducted for an experiment. In the first trial, adults of Tribolium confusum Jacquelin du Val, the confused flour beetle, were exposed on concrete arenas and treated with a combination aerosol of pyrethrin + methoprene dispensed for 5, 10, and 20 min at particle sizes of 4, 8, 12, and 16 μm, inside an aerosol exposure chamber. Nearly all adult T. confusum were knocked down when immediately removed from the chamber. Among all the exposure time and particle size combinations, recovery increased as the post-exposure holding period increased from one to seven days and when adults were transferred to untreated dishes with flour. A second experiment evaluated the residual effect of the aerosol on concrete arenas at 1, 3, and 6 weeks using 3-4-week-old larvae of T. confusum. Adult emergence of exposed larvae decreased with increasing particle size and exposure time. A biological index that assessed development of exposed larvae to the pupal and adult stages was also related to particle size and exposure interval, and this index was correlated with adult emergence. A third set of experiments investigated effects of particle size on adult fecundity for the 10-min exposure time. Male and female adults were cross-mated: exposed female with exposed male, exposed female with unexposed male, exposed male with unexposed female, and unexposed female and unexposed male. Progeny production was reduced as particle size increased, and there were indications that females were affected more than males by the aerosol treatment. This research could be used to improve insect pest management programs by adjusting application equipment to dispense aerosols at particle sizes that give optimum control of exposed adults and residual control of immatures and would also benefit pest management programs.

Insecticidal contact toxicity of several essential oils against stored product pests: Poster

Results of laboratory bioassays in Petri dishes on evaluation of contact toxicity of Illicium verum, Artemisia absinthium and Abies sibirica essential oils (EOs) against larvae of khapra beetle, Trogoderma granarium Ev., adults of grain weevil, Sitophilus granarius L., and rice weevil, Sitophilus oryzae L., and confused flour beetle, Tribolium confusum Duv., and larvae and adults of the lesser mealworm, Tenebrio molitor L., are presented. EOs commercial samples from retail pharmacy were tested at doses 0.01, 0.25, 0.50, 0.75 and 1.00 µl/cm2. A treated Petri dish surface treted with acetone was used as a control. The experiment was carried out in triplicate. Mortality of insects was assessed after 1, 3, 6 and 24 hours post exposure. After exposure insects were placed into untreated Petri dishes for 3 days. The main components of the A. absinthium EOs are thujil alcohol (19.65%), phellandrene (16.71%), borneol (12.1%) and thujone (11.55%) was found. The major component of I. verum EOs was anethole (98.64%). Isobornyl acetate (57.25%), a-pinene (13.55%) and limonene (10.62%) were found as the main components of A. sibirica EOs. S. oryzae and S. granarius were most sensitive to each EO. I. verum EOs was the most effective and caused 100% mortality of each insect at the dose 0.25 µl/cm2.

In Vitro Anthelmintic Activity of Four Plant-Derived Compounds against Sheep Gastrointestinal Nematodes

By using the egg hatch test (EHT), the larval development test (LDT) and the larval mortality/paralysis test (LMT), the in vitro anthelmintic activity on sheep gastrointestinal strongyles (GIS) of four plant-derived pure compounds, mangiferin (at 0.25%, 0.125% and 0.0625%), rutin (at 1%, 0.75%, 0.5%), quercetin (at 1%), and β-sitosterol (at 1%, 0.75%, 0.5%), was investigated. For comparison, untreated and treated (0.1% thiabendazole, 0.1% TBZ) controls were used. Six repetitions were made throughout the experiment. Data were statistically elaborated using the χ2 test. The concentration able to inhibit the development of the 50% of L1s to L3s and causing the mortality of the 50% of L3s (EC50) was also calculated. L3s recovered from untreated Petri dishes were identified at the genus level. In EHT, all tested compounds at all concentrations significantly (p < 0.01) inhibited the hatch of the eggs when compared to the untreated controls, but none of them was as effective as 0.1% TBZ. In LDT, rutin (at 1%, 0.75% and 0.5%), mangiferin (at 0.25% and 0.125%), β-sitosterol (at 1%) and 0.1% TBZ completely prevented the larval development from L1 to L3 in respect to the untreated controls (p < 0.01). In LMT, all tested compounds significantly (p < 0.01) increased the death of L3s compared to the untreated controls, except for β-sitosterol at 0.5%. However, only rutin at all concentrations and 0.25% and 0.125% mangiferin were as effective as 0.1% TBZ. Haemonchus, Trichostrongylus, Chabertia and Teladorsagia/Ostertagia GIS genera, were identified.

From lethality to mortality: exploring the “grey area” of knockdown as an efficacy indicator of different insecticides against major storage insects using a lethality index

In the present work, a lethality index was applied for the evaluation of three insecticides with different modes of action, i.e., thiamethoxam, chlorfenapyr and lambda-cyhalothrin, as surface treatment against adults of Sitophilus oryzae (L.), Oryzaephilus surinamensis (L.) and Tribolium confusum Jacquelin du Val, in laboratory bioassays. The insecticides were applied at their highest label dose, while insect knockdown and mortality were evaluated after 1, 3, 5 and 7 days of exposure to the treated surfaces according to the Standardized Lethality Index, by ranking each insect from “0” to “4,” with “0” corresponding to insects moving normally and “4” corresponding to dead individuals. After the end of the exposure, the surviving insects were transferred to clean, untreated dishes and delayed mortality was assessed after an additional period of 1, 3, 5 and 7 days. Based on the results, initial knockdown was high for S. oryzae after exposure to thiamethoxam and lambda-cyhalothrin even after 1 day of exposure, whereas for longer exposures high mortality levels were recorded. In contrast, in the case of chlorfenapyr knockdown was low after 1 day of exposure; however, high mortality levels were noted after 3 and 5 days of exposure. Similar results were noted for O. surinamensis, which was highly susceptible to all three insecticides tested. The most tolerant insect species to the tested insecticides was T. confusum, since a noticeable number of T. confusum adults were still active at the end of the exposure (7 days). Our results indicate that knockdown is most likely to lead to mortality than to recovery, but there were important differences among the three insecticides tested.

Evaluation of Carifend®, an alpha-cypermethrin-coated polyester net, for the control of Lasioderma serricorne and Ephestia elutella in stored tobacco

The use of insecticide-treated nets has lately attracted scientific interest for the management of mosquitoes and related species, as well as for crop pest control and the protection of stored agricultural commodities. In our study, the effect of Carifend®, an alpha-cypermethrin-coated polyester net, on adults of the two most important stored-tobacco insect pests, Lasioderma serricorne and Ephestia elutella, was evaluated in laboratory bioassays. Carifend® was applied to the bottom of plastic Petri dishes, whereas an additional series of dishes with net material without insecticide was also prepared and served as control. Twenty beetles or moths of mixed sex were placed in each dish. Insects were exposed to the substrates for 1, 5, 15, 30, 60, 90, 120 and 240 min and for 12 and 24 h. Immediate mortality and knockdown effect were measured after each exposure. Afterward, all surviving individuals were placed in untreated plastic dishes and delayed mortality, as well as percentage of knocked down insects, were counted after 1, 3, 5 and 7 days. Immediate mortality was very low for both species tested, even after 24 h of exposure. In contrast, immediate knockdown was high, especially for exposure intervals longer than 60 min. After insect removal from dishes with Carifend®, the high knockdown percentages were gradually transformed in most cases to delayed mortality. Based on our results, Carifend® can provide a satisfactory level of protection for stored tobacco against these two major tobacco insect pests, at least at the conditions tested here.

Contact Toxicity of Deltamethrin Against Tribolium castaneum (Coleoptera: Tenebrionidae), Sitophilus oryzae (Coleoptera: Curculionidae), and Rhyzopertha dominica (Coleoptera: Bostrichidae) Adults.

This study was conducted at Oklahoma State University, Stillwater, OK, to evaluate the response to deltamethrin concentrations for adults of three stored-product insects, Tribolium castaneum (Herbst), Sitophilus oryzae (L.), and Rhyzopertha dominica (F.). In insect toxicological studies, knockdown is the state of intoxication and partial paralysis as a result of exposure to an insecticide. Deltamethrin concentrations ranging from 0.48 to 140 mg/m(2) (1 to 3,000 ppm) were sprayed on glass Petri dishes. After the dishes dried, 20 adult insects of each species were placed on the treated dishes to determine the contact toxicity of deltamethrin. Assessments for knockdown were made at 15-min intervals for up to 8 h after initial exposure and then again after 24 or 48 h. Insects were then transferred to clean untreated Petri dishes with diet and observed from 0.5 to 72 h. Mortality was assessed 72 h after transfer to untreated dishes with food material. Deltamethrin was highly effective against all three species tested and achieved 99% knockdown of insects of all species within 4 h after exposure at concentrations ≥1.2 mg/m(2) Although some insects recovered from initial knockdown at concentrations ≤48 mg/m(2), nearly all the insects were killed at 140 mg/m(2) when exposed for 48 h. LC95 values for all species tested, for the 48-h exposure period, were <140 mg/m(2), the concentration of deltamethrin that could potentially be present in new ZeroFly Storage Bag fabric. ZeroFly bags are used for stored-product insect pest control.

Efficacy of two formulations of pirimiphos-methyl as surface treatment against Sitophilus granarius, Rhyzopertha dominica, and Tribolium confusum

In the present work, the insecticidal effect of two pirimiphos-methyl formulations [emulsifiable concentrate (EC) and capsule suspension (CS)] as surface treatment on concrete, plywood, galvanized metal, plastic, and ceramic tile was evaluated in laboratory bioassays against Sitophilus granarius (L.), Rhyzopertha dominica (F.), and Tribolium confusum Jacquelin du Val. Pirimiphos-methyl was applied at two-dose-rates, 0.01 and 0.1 mg/cm2, and insect mortality was assessed after 1, 2, 3, 4, 5, 6, and 7 days of exposure. After the final mortality count, alive individuals were transferred to untreated dishes, and delayed mortality was evaluated after an additional period of 7 days. In another series of laboratory bioassays, the residual efficacy of these two pirimiphos-methyl formulations on concrete was evaluated against S. granarius, R. dominica, and T. confusum. Pirimiphos-methyl was applied at two-dose-rates, 0.01 and 0.1 mg/cm2, on concrete dishes which were either kept in continuous darkness or exposed to light. A bioassay was initiated at the day of storage and every 4 weeks for 4 consecutive months (5 bioassays in total). Mortality of the exposed adult beetles was measured after 3 and 7 days of exposure. Both formulations were highly effective, with S. granarius being the most susceptible species. In general, mortality of the exposed beetles was not significantly affected by the surface type, with the exception of the significantly lower mortality counts of S. granarius adults on ceramic tile. CS appeared to be much more persistent on concrete than EC for all three species tested. Moreover, CS was not affected by light. In contrast, in some of the combinations tested, EC performance was poor in dishes that were exposed to light, in comparison with dishes that were in continuous darkness. We conclude that the surface type does not significantly and consistently affect S. granarius, R. dominica, and T. confusum mortality after surface application of the two pirimiphos-methyl formulations tested. Furthermore, CS can provide long-term protection when applied on concrete surfaces.

Effects of Natural Cartilaginous Extracellular Matrix on Chondrogenic Potential for Cartilage Cell Transplantation

Autologous chondrocyte transplantation (ACT) has been established to contribute cartilage regeneration over the past years; however, many obstacles need to be overcome. Recently, newer ACT technique involves cotransplantation of chondrocytes and biomaterial. Although various proposed intelligent biomaterials exist, many of them remain insufficient and controversial. In this study, we aimed to examine the effects of natural extracellular matrix (ECM) to the proliferation rate and differentiation on the chondrocytes. We first derived a natural ECM sheet from 10-μm-thick frozen sections of porcine knee cartilages. We then cultured the chondrocytes derived from a rabbit's knee on a dish precoated with the natural ECM. Then we assessed differentiation and chondrogenic potential of the cells compared with those grown in untreated culture dishes. We characterized the gene expression of chondrogenic markers, such as collagen type II, SOX-9, and aggrecan, as well as the level of ECM protein with the use of reverse-transcription polymerase chain reaction analysis. The cells cultured with the ECM sheet showed highest chondrogenic potential and differentiation. Therefore, we can induce good chondrogenesis by with the use of a natural ECM sheet on the culture dish. The readily available and easy-to-handle thin ECM sheets create an environment that promotes efficient cartilage regeneration. Our data suggest that this natural ECM scaffold improved the chondrogenic differentiation of the cells in vitro by providing a favorable microenvironment.

Adult stem cells: potential implications for perioperative medicine

Since their initial description by Friedenstein et al. over 40 years ago, mesenchymal stromal cells (MSCs) have been referred to by a number of terms, including mesenchymal stem cells and ‘‘adult stem cells’’. Friedenstein’s pioneering studies of MSCs described their adherence to untreated culture dishes, their spindle-shaped morphology, as well as their ability to differentiate into chondrocytes, adipocytes, and osteocytes under certain conditions. Although these basic criteria continue to define MSCs, our understanding of these cells has greatly evolved. Of importance, MSCs, unlike embryonic stem cells, are not a single cell type capable of differentiating into multiple cell types in vivo; instead, MSCs are a heterogeneous population of cells capable of a broad number of biological activities. Thus, it has been suggested that the term, ‘‘stromal cell’’, be applied to MSCs as they do not display prototypical stem cell characteristics. Initially, MSCs were isolated from adult bone marrow (where they are believed to support hematopoiesis) as well as adipose tissue and Wharton’s jelly from umbilical cords. These sources remain the best characterized; however, MSCs are now regarded to be ubiquitous cells found in virtually every tissue studied. Controversy exists as to the physiological role of these cells under native in vivo conditions; however, it has been suggested that MSCs may be perivascular cells (‘‘pericytes’’) that support vascular homeostasis and tissue repair. In preclinical studies, perivascular MSCs appear to be mobilized into the circulation under inflammatory conditions, such as traumatic lung injury or hypoxia. A similar mobilization of MSCs into the circulation has also been observed clinically in elderly patients following traumatic hip fracture.

Immobilization of Silk Fibroin as Scaffold for Cell Culture by Plasma Grafting Polymerization

According to the low efficiency of cell attachment and proliferation on commercial polystyrene (PS) dish, scaffold with porous structure on the polystyrene dish is required to improve the cell attachment and proliferation efficiency on the dish. The scaffold with porous structure was fabricated from a solution of powdered silk fibroin by plasma grafting polymerization technique. Argon plasma was utilized by a 13.56 MHz capacitively coupled discharge (CCP) reactor at working pressure of 100 mTorr. Rf power, plasma treatment time, and the cycle of grafting on the dish were varied. The proper treatment time and rf power set to 10 minutes and 100 W, respectively. The experimental results showed the uniformly and highly distributed of porosity of fibroin scaffold on the PS dish surface. The Ar-treated dish had lower UV-Vis absorbance than the untreated dish indicating the efficiency of grafting between fibroin structure and treated PS surface is better than the untreated surface. The absorbance spectra of phanyalanine at 280 nm affirmed the success of the fibroin amino acid grafting to PS aromatic structure.

Rapid prototyping of concave microwells for the formation of 3D multicellular cancer aggregates for drug screening.

Microwell technology has revolutionized many aspects of in vitro cellular studies from 2D traditional cultures to 3D in vivo-like functional assays. However, existing lithography-based approaches are often costly and time-consuming. This study presents a rapid, low-cost prototyping method of CO2 laser ablation of a conventional untreated culture dish to create concave microwells used for generating multicellular aggregates, which can be readily available for general laboratories. Polymethylmethacrylate (PMMA), polydimethylsiloxane (PDMS), and polystyrene (PS) microwells are investigated, and each produces distinctive microwell features. Among these three materials, PS cell culture dishes produce the optimal surface smoothness and roundness. A549 lung cancer cells are grown to form cancer aggregates of controllable size from ≈40 to ≈80 μm in PS microwells. Functional assays of spheroids are performed to study migration on 2D substrates and in 3D hydrogel conditions as a step towards recapitulating the dissemination of cancer cells. Preclinical anti-cancer drug screening is investigated and reveals considerable differences between 2D and 3D conditions, indicating the importance of assay type as well as the utility of the present approach.

Localized treatments using commercial dust and liquid formulations of fipronil against Coptotermes formosanus (Isoptera: Rhinotermitidae) in the laboratory

Use of proper application methods and formulations of termiticides are important to reduce their negative impact to the environment. In this study, we conducted laboratory experiments to determine the effect of localized treatments with commercial dust and liquid formulations of fipronil against Formosan subterranean termites, Coptotermes formosanus Shiraki. The test arena consisted of a specially designed 16-chambered structure with a center chamber connected to 5 foraging chambers that themselves were connected to 10 additional foraging chambers. One peripheral chamber received a liquid or dust treatment and termites were released in the center chamber. Results showed that >91% of the termites were dead within the 9-d test period despite the localized treatment of only 1 foraging chamber. Termites that were still alive after 9 d were transferred to an untreated dish and held for 10 more days. The majority of those termites were dead and the rest were moribund on day 19. Regardless of the specific dish treated, both formulations of fipronil were found to be highly efficacious. Termites did not exhibit repellency to either formulation. Our results suggest that localized (or spot) treatment with either commercially available dust or liquid formulations of fipronil can be a viable option for control of a termite infestation where complete soil drenching is not desirable.

Evaluation of biological control agents for Fusarium wilt in Hiemalis begonia

Five laboratory-isolated Bacillus subtilis strains: CB1, CGB, JHB1, SSB1 and WW4; and five commercial biocontrol agents: Actinovate® (Stretomyces lydicus strain WYEC108), Serenade® MAX (Bacillus subtilis strain QST713), Mycostop® (Streptomyces griseoviridis strain K61), RootShield® (Trichoderma harzianum strain T-22), and Prestop® WP (Gliocladium catenulatum strain J1446) were evaluated for their potential to inhibit Fusarium foetens both in vitro in Petri dishes and in the greenhouse using potted begonia plants. Bacillus subtilis strain JHB1 inhibited F. foetens mycelial growth by 76% compared with the untreated dishes after seven days of dual culturing; and also provided significant protection to potted begonia plants against F. foetens under greenhouse conditions (two trials) when applied as two soil drenches prior to pathogen inoculation. Compared with the pathogen-inoculated control, disease severity (DS) was reduced by 33% and 50% by JHB1 in trial one and two, respectively. All of the commercial biocontrol agents significantly reduced DS compared with the control when applied as soil drenches prior to pathogen inoculation. Most of the commercial products delayed plant disease symptom development by seven or more days. Moreover, Mycostop, Prestop, Actinovate, RootShield and Serenade all significantly reduced the DS, and increased the shoot dry weights and chlorophyll content index for over 28 days after inoculation compared with the control.

Efficacy of a Combination of Beta-Cyfluthrin and Imidacloprid and Beta-Cyfluthrin Alone for Control of Stored-Product Insects on Concrete

The insecticidal effect of Temprid, a formulation that contains beta-cyfluthrin and imidacloprid, was tested on concrete for control of seven stored-product insect species: the rusty grain beetle, Cryptolestes ferrugineus (Stephens); the sawtoothed grain beetle, Oryzaephilus surinamensis (L.); the red flour beetle, Tribolium castaneum (Herbst); the confused flour beetle, T. confusum Jacquelin du Val; the hide beetle, Dermestes maculatus (DeGeer); and the psocids Liposcelis bostrychophila Badonnel and L. paeta Pearman. Temprid, which contains 10.5% beta-cyfluthrin and 21% imidacloprid, was tested at the rate of 17.2 ml of formulation/4.1 liters of water/100 m2. Adults were exposed for 4, 8, 14, and 24 h, and then daily for 7 d, on untreated dishes or dishes treated with Temprid. In the untreated dishes, mortality of C. ferrugineus and O. surinamensis was lower when food was present, but food did not affect mortality of the other species. Presence of food did not affect mortality of any of the species tested in the treated dishes. C. ferrugineus, O. surinamensis, and the two psocid species were very susceptible to Temprid, with mortality of 97-100% after 7 d of exposure. In contrast, D. maculatus, T. castaneum, and T. confusum were tolerant to Temprid, as mortality did not exceed 57, 25, and 17%, respectively, at the 7-d exposure. A separate series of similar bioassays with Tempo, a formulation that contains 11.8% beta-cyfluthrin alone, which was applied at the same dose rate as Temprid, was conducted using O. surinamensis and T. castaneum as the target insect species, and results showed that Tempo was at least as effective as Temprid. Our results indicate that the simultaneous use of beta-cyfluthrin with imidacloprid is not more effective on concrete than beta-cyfluthrin alone, and efficacy of both formulations varies with the target species.

Evaluating the performance of fibrillar collagen films formed at polystyrene surfaces as cell culture substrates

While it is well-appreciated that the extracellular matrix plays a critical role in influencing cell responses, well-defined and reproducible presentation of extracellular matrix proteins poses a challenge for in vitro experiments. Films of type 1 collagen fibrils assembled on alkanethiolate monolayers formed at gold-coated surfaces have been shown to elicit a cellular response comparable to collagen gels, but with the advantages of excellent optical properties, and high reproducibility and robustness. To make this collagen matrix more accessible to laboratories that do not have access to gold film deposition the authors have examined the use of untreated polystyrene as a substrate for forming fibrillar collagen films. Direct comparison of films of fibrillar collagen fibrils formed at polystyrene with those formed at alkanethiolate monolayers indicates that films of collagen formed on these two surfaces compare very favorably to one another, both in their supramolecular structural characteristics as well as in the cell response that they elicit. Both substrates exhibit a dense covering of fibrils approximately 200 nm in diameter. The spreading of fibroblasts and activation of the tenascin-C gene promoter are statistically equivalent as determined by a metric derived from the D-statistic normally used in the Kolmogorov-Smirnov statistical test. The results of this study suggest that biologically relevant, robust thin films of collagen fibrils can be formed in any laboratory in untreated polystyrene dishes and multi-well polystyrene plates.

Neuregulin 1β/erbB signalling in cardiac myocytes enhances integrin based myocyte‐matrix interactions via src

The Neuregulin‐1β (Nrg)‐erbB2/ErbB4 (E2/E4) system is critical for the heart and it is needed for myocyte survival in stress conditions. Nrg activates Src and FAK, both part of integrin signaling. Hyp.: Nrg signaling regulates myocyte matrix interactions, critical for coupling force generation to the extracellular matrix.Immunostaining of mouse hearts showed E4 and E2 are at intercalated discs and have a t‐tubule like striated pattern. Isolated myocytes were incubated 30 min or 24h with an E2 monoclonal antibody (Ab) or an E2 tyrosine kinase inhibitor (TKI). Western‐blot showed E2 TKI blocks activation of FAK, Erk1/2, and Akt by Nrg; Ab is a weak inhibitor. ErbB activation in myocyte‐matrix coupling was tested by plating myocytes onto untreated culture dishes spot‐coated with laminin, with or without Nrg (10ng/ml). After 30 minutes unattached cells were stained and counted. Attachment was enhanced 4‐fold by Nrg vs. laminin alone. Cells did not attach to Nrg coated plates in absence of laminin. Pre‐treatment of myocytes with E2 inhibitors did not prevent Nrg‐enhanced attachment; pre‐treated with Src inhibitor PP2 (10 μM) Nrg effect was inhibited.Nrg‐1β treatment of cardiac myocytes enhances integrin mediated myocyte‐matrix coupling via Src dependent signalling. This appears to be E2 independent pathway, involving E4 signaling, supported by e4 localization to the intercalated disks in vivo.Support by SNSF, NIH.