Standardizing Immunohistochemistry: A New Reference Control for Detecting Staining Problems

A new standardized immunohistochemistry (IHC) control for breast cancer testing comprises formalin-fixed human epidermal growth factor receptor 2, estrogen receptor, or progesterone receptor peptide antigens covalently attached to 8-µm glass beads. The antigen-coated beads are suspended in a liquid matrix that hardens upon pipetting onto a glass microscope slide. The antigen-coated beads remain in place through deparaffinization, antigen retrieval, and immunostaining. The intensity of the beads’ stain provides feedback regarding the efficacy of both antigen retrieval and immunostaining. As a first report, we tested the sensitivity and specificity of the new IHC controls (“IHControls”). To evaluate sensitivity, various staining problems were simulated. IHControls detected primary and secondary reagent degradation similarly to tissue controls. This first group of IHControls behaved similarly to tissue controls expressing high concentrations of the antigen. The IHControls were also able to detect aberrations in antigen retrieval, as simulated by sub-optimal times or temperatures. Specificity testing revealed that each antigen-coated bead was specific for its cognate IHC test antibody. The data support the conclusion that, like tissue controls, IHControls are capable of verifying the analytic components of an immunohistochemical stain. Unlike tissue controls, IHControls are prepared in large bulk lots, fostering day-to-day reproducibility that can be standardized across laboratories.     

Use of two formulations and two application techniques to deliver Bacillus thuringiensis var. israelensis for the control of Musca domestica (Diptera: Muscidae) larvae and adults in poultry houses

A field study was carried out for 6 wks to assess, from both an efficiency and economic perspective, the effect of individual and integrated success of feeding and topical applications of two formulations of Bacillus thuringiensis var. israelensis (Bti) in controlling house fly (Musca domestica L.) larvae and adults in poultry houses. There was no significant difference between the 1 g and 2 g L^?1 spray applications of Bti. In the absence of spray applications, no significant differences in larval mortalities were observed between the 250 mg and 500 mg kg^?1 feed applications. The percentage mortality of larvae accomplished as a result of using a combination of 250 mg kg^?1Bti feed and 2 g L^?1 spray applications was equivalent to that obtained as a result of combining 500 mg kg^?1Bti and 1g L^?1 spray applications. Treatment with Bti caused significant reductions in the emergence (up to 74%) of house fly adults compared to the control. The fact that the emergence of adult house flies was affected by Bti treatments implies that Bti has sublethal effects on house fly larvae. The cost–benefit analysis (expressed in terms of mortality of larvae growing) indicated that the most effective combination for house fly larvae and adult house fly emergence control was the 500 mg kg^?1 of feed and 2 g L^?1 spray application combination that resulted in 67% larval mortality and a 74% decrease in adult house fly emergence. This study presents commercial users with various alternatives for possible combinations of the two Bti formulations.     

Current levels of suppression of waterhyacinth in Florida USA by classical biological control agents

Waterhyacinth, Eichhornia crassipes (Mart.) Solms (Pontederiaceae), has been a global target for classical biological control efforts for decades. In Florida, herbicidal application is the primary control method employed, usually without regard for the activities of the three biological control agents introduced intentionally during the 1970s, namely Neochetina eichhorniae Warner, Neochetina bruchi, Hustache (Coleoptera: Curculionidae), and Niphograpta albiguttalis Warren (Lepidoptera: Crambidae). A series of field experiments from 2008 to 2010 was conducted at four Florida sites using an insecticide-check approach to quantify the current levels of suppression provided by these agents. In the field N. albiguttalis was rarely found while more than 99% of all Neochetina sp. adults were N. eichhorniae. Although it was not possible to disentangle the relative impacts of Neochetina sp. adults from larvae on individual plant variables, the larvae played a major role in reducing plant biomass and the number of inflorescences. Plots exposed to unrestricted herbivory contained 58.2% less biomass and produced 97.3% fewer inflorescences at the end of the experiments. Despite these large reductions, herbivory decreased waterhyacinth coverage by only 16.8% and most of this was attributed to a low-nutrient site where coverage was reduced disproportionately. Overall, coverage trended upwards during the course of the experiments and was always close to 100% when the plots were harvested. Although coverage is a somewhat arbitrary metric, especially for floating plants subject to compression and dispersion, it influences the perception of biological control efficacy which, in turn, directly influences herbicide management decisions in Florida. Despite waterhyacinth populations that now produce less than half as much biomass and up to 98% fewer seeds than before the deployment of biological control agents, the overall approach used to achieve maintenance control of the plant in Florida will probably not change unless new biological control agents, such as Megamelus scutellaris Berg (Hemiptera: Delphacidae), can reduce coverage significantly.     

Alginate embedding and subsequent sporulation of in vitro-produced Conidiobolus thromboides hyphae using a pressurised air-extrusion method

Conidiobolus thromboides is an entomophthoralean fungus with potential as a biological control agent of aphids. However, its application in biological control is limited due to its formulation requirements. The objective of this study was to develop and optimise a novel air-extrusion method to embed C. thromboides hyphae at high density in alginate pellets. An orthogonal experimental design was used to investigate selected combinations of parameters known to affect hyphal density within pellets. The diameter of pellets produced, and the calculated density of hyphae within them, ranged from 0.18 ± 0.09 to 3.17 ± 0.06 mm and from 0.02 to 350.56 mg/mm³ respectively. These data were used to predict the optimal parameter combination to deliver the greatest density of hyphae of C. thromboides per pellet: 1% sodium alginate, a 1:2 ratio of hyphae to sodium alginate, an orifice diameter of 0.232 mm and an air pressure of 0.05 MPa. Pellets made under the optimal conditions predicted produced a mean total of 4.3 ± 0.6 × 10⁵ conidia per pellet at 100% relative humidity which was significantly greater than the mean total number of conidia produced from infected aphid cadavers of comparable size (9.35 ± 0.85 × 10⁴) (p < 0.001). In conclusion, air-extrusion embedding appears to be a promising method for formulating in vitro-produced hyphae of C. thromboides for use in biological control.     

Serotonin transporter gene,childhood emotional abuse and cognitive vulnerability to depression

Meta‐analyses evaluating the association between the serotonin transporter polymorphism (5‐HTTLPR) with neuroticism and depression diagnosis as phenotypes have been inconclusive. We examined a gene–environment interaction on a cognitive vulnerability marker of depression, cognitive reactivity (CR) to sad mood. A total of 250 university students of European ancestry were genotyped for the 5‐HTTLPR, including SNP rs25531, a polymorphism of the long allele. Association analysis was performed for neuroticism, CR and depression diagnosis (using a self‐report measure). As an environmental pathogen, self‐reported history of childhood emotional abuse was measured because of its strong relationship with depression. Participants with the homozygous low expressing genotype had high CR if they had experienced childhood emotional maltreatment but low CR if they did not have such experience. This interaction was strongest on the Rumination subscale of the CR measure. The interaction was not significant with neuroticism or depression diagnosis as outcome measures. Our results show that 5‐HTTLPR is related to cognitive vulnerability to depression. Our findings provide evidence for a differential susceptibility genotype rather than a vulnerability genotype, possibly because of the relatively low levels of abuse in our sample. The selection of phenotype and environmental contributor is pivotal in investigating gene–environment interactions in psychiatric disorders.     

The effects of cell density and metabolite flux on cellular dynamics

Summary Density-dependent regulation of cell growth in tissue culture is a well-known phenomenon but the mechanism of regulation remains obscure. Here we explore the effects of cell density and metabolite flux on the collective dynamics of a cell population. The intracellular dynamics are modelled by positive feedback kinetic mechanisms of the kind known to apply to yeast cells. Several experimental observations related to glycolytic oscillations are predicted and it is suggested that the general conclusions may be applicable in a broader context.     

Performance of Two Fruit Fly (Diptera: Tephritidae) Pupal Parasitoids (Coptera haywardi [Hymenoptera: Diapriidae] and Pachycrepoideus vindemiae [Hymenoptera: Pteromalidae]) under Different Environmental Soil Conditions

We evaluated the performance of Coptera haywardi (Ogloblin) (Diapriidae) and Pachycrepoideus vindemiae (Rondani) (Pteromalidae), both hymenopteran pupal parasitoids of Anastrepha spp. (Diptera: Tephritidae). Performance was studied by manipulating the following environmental conditions in the laboratory: (1) soil type, (2) soil moisture content, (3) soil compaction, and (4) depth at which pupae were buried in the soil. There were two experiments: in the first, exposure time of pupae was held constant and in the second, it varied. In the first experiment, C. haywardi was significantly more effective than P. vindemiae in parasitizing fly pupae. With exposure time held constant (36 h), only soil type and pupal burial depth were significantly related to parasitism rates. While P. vindemiae only parasitized pupae located on the soil surface, C. haywardi attacked pupae that were buried up to 5 cm deep, performing better in clayey than in loamy soil. In the second experiment, exposure time (24, 36, 48, and 72 h) had no significant effect on parasitism rates, but soil type did. P. vindemiae again only attacked pupae on the soil surface while C. haywardi was also able to parasitize pupae that were buried up to 5 cm deep. We conclude that C. haywardi represents a viable candidate to replace the environmentally unfriendly P. vindemiae in augmentative biological control programs against fruit flies.     

In vivo control of Mycosphaerella pinodes on pea leaves by a crude bulb extract of Eucomis autumnalis

We previously reported on the in vitro antifungal activity of a crude whole plant extract from Eucomis autumnalis against seven economically important plant pathogenic fungi. A crude extract of the bulb showed similar in vitro mycelial growth inhibition of the same plant pathogenic fungi as well as that of an eighth fungus, Mycosphaerella pinodes, the cause of black spot or Ascochyta blight, in peas. Subsequently, fourth internode leaves were removed from 4 wk old pea plants, placed on moist filter paper in Petri dishes and inoculated with an M. pinodes spore suspension before and after treatment with the extract. The control of Ascochyta blight by different concentrations of the crude E. autumnalis extract was followed in vivo by leaf symptoms over a 6 day period at 20°C in a growth cabinet. The crude extract prevented M. pinodes spore infection of the leaves when the leaves were inoculated with spores both before or after treatment with the extract, confirming complete inhibition of spore germination. The crude E. autumnalis extract showed no phytotoxic reaction on the leaves even at the highest concentration applied.