Cows as Bioreactors for the Production of Nutritionally and Biomedically Significant Proteins.

Dairy and beef cattle make a vital contribution to global nutrition, and since their domestication, they have been continuously exposed to natural and artificial selection to improve production characteristics. The technologies of transgenesis and gene editing used in cattle are responsible for generating news characteristics in bovine breeding, such as alteration of nutritional components of milk and meat enhancing human health benefits, disease resistance decreasing production costs and offering safe products for human food, as well as the recombinant protein production of biomedical significance. Different methodologies have been used to generate transgenic cattle as bioreactors. These methods include the microinjection of vectors in pronuclear, oocyte or zygote, sperm-mediate transgenesis, and somatic cell nuclear transfer. Gene editing has been applied to eliminate unwanted genes related to human and animal health, such as allergy, infection, or disease, and to insert transgenes into specific sites in the host genome. Methodologies for the generation of genetically modified cattle are laborious and not very efficient. However, in the last 30 years, transgenic animals were produced using many biotechnological tools. The result of these modifications includes (1) the change of nutritional components, including proteins, amino acids and lipids for human nutrition; (2) the removal allergic proteins milk; (3) the production of cows resistant to disease; or (4) the production of essential proteins used in biomedicine (biomedical proteins) in milk and blood plasma. The genetic modification of cattle is a powerful tool for biotechnology. It allows for the generation of new or modified products and functionality that are not currently available in this species.

Invasion of Wolbachia at the residential block level is associated with local abundance of Stegomyia aegypti, yellow fever mosquito, populations and property attributes.

Wolbachia can suppress dengue and control mosquito populations and this depends on the successful invasion of Wolbachia-infected mosquitoes into local populations. Ovitrap data collected during the recent invasion of wMel-infected Stegomyia aegypti (Diptera: Culicidae) (Linnaeus) into Gordonvale near Cairns, Australia, were used to identify variables that help predict the success of localized invasion. Based on the variance in Wolbachia frequencies across Gordonvale as well as at another release site at Yorkeys Knob in comparison to simulations, it was estimated that on average 2-4 females contributed eggs to an ovitrap. By collating ovitrap data from two collection periods at the start of the release from residential blocks, it was found that uninfected mosquitoes had a patchy distribution across the release site. Residential blocks with relatively high uninfected mosquito numbers were less easily invaded by Wolbachia than blocks with low numbers. The numbers of uninfected mosquitoes in ovitraps were negatively correlated with the proportion of brick houses in a residential block, whereas local Wolbachia frequencies were correlated positively with this variable as well as negatively with the amount of shading in a yard and availability of breeding sites. These findings point to proxy measures for predicting the ease of localized invasion of Wolbachia.

Evaluation of in vivo T cell kinetics: use of heavy isotope labelling in type 1 diabetes.

CD4(+) memory cell development is dependent upon T cell receptor (TCR) signal strength, antigen dose and the cytokine milieu, all of which are altered in type 1 diabetes (T1D). We hypothesized that CD4(+) T cell turnover would be greater in type 1 diabetes subjects compared to controls. In vitro studies of T cell function are unable to evaluate dynamic aspects of immune cell homoeostasis. Therefore, we used deuterium oxide ((2) H(2)O) to assess in vivo turnover of CD4(+) T cell subsets in T1D (n = 10) and control subjects (n = 10). Serial samples of naive, memory and regulatory (T(reg)) CD4(+) T cell subsets were collected and enrichment of deoxyribose was determined by gas chromatography-mass spectrometry (GC-MS). Quantification of T cell turnover was performed using mathematical models to estimate fractional enrichment (f, n = 20), turnover rate (k, n = 20), proliferation (p, n = 10) and disappearance (d*, n = 10). Although turnover of T(regs) was greater than memory and naive cells in both controls and T1D subjects, no differences were seen between T1D and controls in T(reg) or naive kinetics. However, turnover of CD4(+) memory T cells was faster in those with T1D compared to control subjects. Measurement and modelling of incorporated deuterium is useful for evaluating the in vivo kinetics of immune cells in T1D and could be incorporated into studies of the natural history of disease or clinical trials designed to alter the disease course. The enhanced CD4(+) memory T cell turnover in T1D may be important in understanding the pathophysiology and potential treatments of autoimmune diabetes.

An autoimmune-associated variant in PTPN2 reveals an impairment of IL-2R signaling in CD4(+) T cells.

The IL-2/IL-2R signaling pathway has an important role in autoimmunity. Several genes identified in genome-wide association (GWA) studies encode proteins in the IL-2/IL-2R signaling cascade that are associated with autoimmune diseases. One of these, PTPN2, encodes a protein tyrosine phosphatase that is highly expressed in T cells and regulates cytokine signaling. An intronic risk allele in PTPN2, rs1893217(C), correlated with decreased IL-2R signaling in CD4(+) T cells as measured by phosphorylation of STAT5 (phosphorylated STAT5 (pSTAT5)). We modeled an additive single nucleotide polymorphism (SNP) genotype, in which each copy of the risk allele conferred a decrease in IL-2R signaling (P=4.4 × 10(-8)). Decreased pSTAT5 impacted IL-2Rß chain signaling resulting in reduced FOXP3 expression in activated cells. This phenotype was not due to overt differences in expression of the IL-2R, molecules in the IL-2R signaling cascade or defects in STAT5. However, the rs1893217(C) risk variant did correlate with decreased PTPN2 expression in CD4(+)CD45RO T cells (P=0.0002). Thus, the PTPN2rs1893217(C) risk allele associated with reduced pSTAT5 in response to IL-2 and reduced PTPN2 expression. Together, these data suggest that decreased expression of PTPN2 may indirectly modulate IL-2 responsiveness. These findings, identified through genotype/phenotype relationships, may lead to identification of novel mechanisms underlying dysregulation of cytokine signaling in autoimmunity.

Australian action to reduce health risks from radon.

In Australia, worker exposure to radon in underground uranium mines has been a focus of policy makers and regulators, and has been well controlled in the industry sector. That cannot be said for public exposure to radon. Radon exposure studies in the late 1980s and early 1990s demonstrated that the levels of radon in Australian homes were some of the lowest in the world. The International Basic Safety Standards, published by the International Atomic Energy Agency, requires the government to establish and implement an action plan for controlling public exposure due to radon indoors. When considering different policy options, it is important to develop radon prevention and mitigation programmes reflecting elements that are unique to the region or country. The Australian Radon Action Plan is being considered at a national level, and presents a long-range strategy designed to reduce radon-induced lung cancer in Australia, as well as the individual risk for people living with high concentrations of radon. In Australia, workers who are not currently designated as occupationally exposed are also considered as members of the public. In the Australian context, there are only a limited set of scenarios that might give rise to sufficiently high radon concentrations that warrant mitigation. These include highly energy efficient buildings in areas of high radon potential, underground workplaces, workplaces with elevated radon concentrations (e.g. spas using natural spring waters), and enclosed workspaces with limited ventilation. The key elements for a successful plan will rely on collaboration between government sectors and other health promotion programmes, cooperative efforts involving technical and communication experts, and partnering with building professionals and other stakeholders involved in the implementation of radon prevention and mitigation.

A lethal ovitrap-based mass trapping scheme for dengue control in Australia: I. Public acceptability and performance of lethal ovitraps.

We report on the first field evaluation of the public acceptability and performance of two types of lethal ovitrap (LO) in three separate trials in Cairns, Australia. Health workers were able to set standard lethal ovitraps (SLOs) in 75 and 71% of premise yards in the wet and dry season, respectively, and biodegradable lethal ovitraps (BLOs) in 93% of yards. Public acceptance, measured as retention of traps by residents, was high for both trap types, with <9% of traps missing after 4 weeks. Traps retaining water after 4 weeks were 78 and 34% for the two SLO trials and 58% for the BLOs. The 'failure rate' in the 535 BLOs set in the field for 4 weeks was 47%, of which 19% were lost, 51% had holes from probable insect chewing, 23% were knocked over, 7% had dried by evaporation and 1% were split. There was no significant difference in the failure rate of BLOs set on porous (grass, soil and mulch) versus solid (tiles, concrete, wood and stone) substrates. The SLOs and the BLOs were readily acceptable to ovipositing Aedes aegypti L. (Diptera: Culicidae); the mean number of eggs/trap was 6 and 15, for the dry season and wet season SLO trial, respectively, and 15 for the BLO wet season trial. Indeed, 84-94% of premise yards had egg positive SLOs or BLOs. A high percentage of both wet and dry season SLOs (29 and 70%, respectively) and BLOs (62%) that were dry after 4 weeks were egg positive, indicating the traps had functioned. Lethal strips from SLOs and BLOs that had been exposed for 4 weeks killed 83 and 74%, respectively, of gravid Ae. aegypti in laboratory assays. These results indicate that mass trapping schemes using SLOs and BLOs are not rejected by the public and effectively target gravid Ae. aegypti. The impact of the interventions on mosquito populations is described in a companion paper.

A lethal ovitrap-based mass trapping scheme for dengue control in Australia: II. Impact on populations of the mosquito Aedes aegypti.

In Cairns, Australia, the impacts on Aedes aegypti L. (Diptera: Culicidae) populations of two types of 'lure & kill' (L&K) lethal ovitraps (LOs), the standard lethal ovitrap (SLO) and the biodegradable lethal ovitrap (BLO) were measured during three mass-trapping interventions. To assess the efficacy of the SLO, two interventions (one dry season and one wet season) were conducted in three discrete areas, each lasting 4 weeks, with the following treatments: (i) SLOs (>200 traps, approximately 4/premise), BG-sentinel traps (BGSs; approximately 15, 1/premise) and larval control (container reduction and methoprene treatment) and (ii) larval control alone, and (iii) untreated control. Female Ae. aegypti populations were monitored for 4 weeks pre- and post-treatment in all three areas using BGSs and sticky ovitraps (SOs) or non-lethal regular ovitraps (ROs). In the dry season, 206 SLOs and 15 BGSs set at 54 and 15 houses, respectively, caught and killed an estimated 419 and 73 female Ae. aegypti, respectively. No significant decrease in collection size of female Ae. aegypti could be attributed to the treatments. In the wet season, 243 SLOs and 15 BGSs killed approximately 993 and 119 female Ae. aegypti, respectively. The mean number of female Ae. aegypti collected after 4 weeks with SOs and BGSs was significantly less than the control (LSD post-hoc test). The third mass-trapping intervention was conducted using the BLO during the wet season in Cairns. For this trial, three treatment areas were each provided with BLOs (>500, approximately 4/premise) plus larval control, and an untreated control area was designated. Adult female Ae. aegypti were collected for 4 weeks pre- and post-treatment using 15 BGSs and 20 SOs. During this period, 53.2% of BLOs contained a total of 6654 Ae. aegypti eggs. Over the intervention period, collections of Ae. aegypti in the treatment areas were significantly less than in the control area for BGSs but not SOs. An influx of relatively large numbers of young females may have confounded the measurement of changes in populations of older females in these studies. This is an important issue, with implications for assessing delayed action control measures, such as LOs and parasites/pathogens that aim to change mosquito age structure. Finally, the high public acceptability of SLOs and BLOs, coupled with significant impacts on female Ae. aegypti populations in two of the three interventions reported here, suggest that mass trapping with SLOs and BLOs can be an effective component of a dengue control strategy.

The composition of the ceramides from human stratum corneum and from comedones.

Human epidermal surface lipids were collected by an ethanol wash and the ceramides were quantified by thin-layer chromatography-photodensitometry. Six ceramide fractions were isolated and the structural components of each were analyzed in detail. The most unusual of the epidermal ceramides contained a sphingosine base with amide-linked 30- and 32-carbon omega-hydroxyacids and an ester-linked nonhydroxyacid, 41% of which was linoleic acid. The proportion of linoleic acid in the analogous ceramide from comedones was 6%. This supports the hypothesis that a localized insufficiency of linoleic acid in the follicular epithelium is an etiologic factor in comedogenesis.

Potent and selective nonpeptide inhibitors of caspases 3 and 7.

5-Dialkylaminosulfonylisatins have been identified as potent, nonpeptide inhibitors of caspases 3 and 7. The most active compound within this series (34) inhibited caspases 3 and 7 in the 2-6 nM range and exhibited approximately 1000-fold selectivity for caspases 3 and 7 versus a panel of five other caspases (1, 2, 4, 6, and 8) and was at least 20-fold more selective versus caspase 9. Sequence alignments of the active site residues of the caspases strongly suggest that the basis of this selectivity is due to binding in the S2 subsite comprised of residues Tyr204, Trp206, and Phe256 which are unique to caspases 3 and 7. These compounds inhibit apoptosis in three cell-based models: human Jurkat T cells, human chondrocytes, and mouse bone marrow neutrophils.

Human stratum corneum polar lipids and desquamation.

The ceramides and steryl-sulfate components from desquamated and cohesive human stratum corneum were examined using a combination of chemical and chromatographic means. Six structurally distinct series of ceramides were identified, and the relative amounts of these species, as measured by quantitative thin-layer chromatography, did not differ in cohesive and desquamated stratum corneum. In contrast, the level of cholesteryl sulfate was significantly reduced in the desquamated material. The results are in accord with the hypothesis that cholesteryl sulfate serves in cell-to-cell cohesion within the stratum corneum, and its hydrolysis may be necessary to permit shedding of cells from the surface.

Acoustic neuroma volume: MRI-based calculations and clinical implications.

This study developed a technique for magnetic resonance imaging (MRI)-based volume calculation and defined the specific volume-diameter relationship in acoustic neuromas (AN). Computer-assisted measurement calculated the area of tumor in each MRI slice. Volume was determined by multiplying area by slice interval. The technique was validated by imaging known volumes of gadolinium. The precision was greater than 95%. Our volume calculations revealed a specific volume-diameter relationship in ANs. The following conclusions can be drawn: 1. our technique can accurately determine AN tumor volume; 2. the AN volume-diameter relationship is a specific function and cannot be predicted by assumptions of tumor shape; 3. the volume-diameter relationship should be combined with clinical data to determine the "break point" at which small increments of diameter produce significant clinical sequelae.

Factors related to quality of life and functional status in 50 patients with head and neck cancer.

Quality of life (QOL) and functional status (FS) have become important outcome measures in cancer therapy. Valid and reliable instruments recently have been developed for examining QOL and FS in patients with head and neck (HN) cancer. The present study evaluated the relationships of QOL and FS to physical and psychological variables assumed to affect QOL and FS. Fifty patients were evaluated up to 6 years after HN cancer surgery using one general QOL instrument and three HN-specific instruments. Analysis of variance showed physical variables such as tumor site to be related to HN-specific scores, while psychosocial variables such as marital status were related to general QOL scores (P < or = .05). Several relationships were seen between physical or psychosocial variables and FS or QOL measures; however the relationships were not as strong or direct as expected.

Relationship between quality of life and depression in patients with head and neck cancer.

OBJECTIVE: This study describes the relationship between health-related quality of life (HRQOL) and depression in patients following major surgery for head and neck cancer. DESIGN: Cross-sectional study using medical chart review, patient interview, and test administration. METHOD: Fifty patients were evaluated 6 months to 6 years following surgery using one global HRQOL measure (The Functional Assessment of Cancer Therapy-General [FACT-G]); three disease-specific measures of HRQOL (the HN module of the FACT [FACT-HNS], The University of Washington Quality of Life Scale [UWQOL], and The Performance Status Scale for Head and Neck Cancer); and one measure of depression (The Beck Depression Inventory [BDI]). RESULTS: The study population showed a high occurrence of depressive symptoms (22%). A negative correlation existed between the BDI and HRQOL as measured by the FACT-G (r = -0.49, P < .001) and the UWQOL (r = -0.44, P = .003). When somatic symptoms of depression were removed, the BDI remained correlated with HRQOL and was most highly correlated with the Emotional Well-Being (EWB) subscale of the FACT-G (r = -0.42, P = .003). There was no correlation between clinician judgments of EWB and any patient-rated measures of HRQOL or depression. CONCLUSIONS: Results demonstrate an inverse relationship between patient-reported HRQOL and depression. The lack of correlation between physician and patient ratings of HRQOL and EWB stresses the importance of obtaining patient ratings in addition to traditional clinician ratings when assessing outcomes. Finally, the multidimensional construction of the FACT with its specific subscales may make it a useful clinical tool for assessing patient status and augmenting patient interviews.

Quality of life and functional status measures in patients with head and neck cancer.

OBJECTIVE: To assess the relationship among three validated head and neck-specific measures of functional status and a general measure of quality of life in patients with head and neck cancer. DESIGNS: Cross-sectional study using medical chart review, patient interview, and test administration. SETTING: Academic tertiary referral center. PARTICIPANTS: Fifty adults patients 3 months to 6 years after major surgery for head and neck cancer. MAIN OUTCOME MEASURE: Scores from a general measure of quality of life (the Functional Assessment of Cancer Therapy), a subscale specific to head and neck cancer, the University of Washington Quality of Life Questionnaire, and the Performance Status Scale for Head and Neck Cancer Patients. RESULTS: The disease-specific measures of functional status correlate well with one another. However, there were low correlations between the Functional Assessment of Cancer Therapy and the disease-specific measures, indicating that general and disease-specific instruments contribute unique information about quality of life. CONCLUSION: A general measure of quality of life augments information obtained by disease-specific instruments by interpreting functional status in the broader scope of the patient's life.

Domoic acid: neurobehavioral and neurohistological effects of low-dose exposure in adult rats.

Adult rats treated IP with domoic acid at 0, 0.22, 0.65, or 1.32 mg/kg were tested for passive avoidance (PA), auditory startle (AS), or conditioned avoidance (CAR) behaviors. Clinical signs were observed only at the 1.32 mg/kg dose level. Within 24 h of dosing, rats surviving a dose of 1.32 mg/kg exhibited transient decreased body weight and exaggerated AS responding. Startle latency and habituation, PA, and CAR were not affected. Examination of brains from six rats per group revealed a subset (2/6) of animals receiving 1.32 mg/kg domoic acid with degenerating neurons in the hippocampal CA1/CA3 subregions and gliosis. The decreased body weight and increased startle suggest a hyperreactivity syndrome possibly related to neuronal degeneration in the hippocampus. In a separate experiment, domoic acid at an IP dose of 0.93 mg/kg was found to produce hypomotility in addition to a decrease in body weight. Both effects were reduced by pretreatment with scopolamine (2 mg/kg), but not with caffeine (30 mg/kg), indicating a possible cholinergic involvement in domoate's toxicity.

Effects of platelet activating factor on vascular permeability of the middle ear mucosa.

Platelet activating factor (PAF), a potent inflammatory mediator, seems to play a significant role in the pathogenesis of otitis media with effusion (OME), along with other inflammatory mediators such as leukotrienes and prostaglandins. The purpose of this study was to investigate the effect of PAF on the vascular permeability of middle ear mucosa, in an experimental OME model using chinchillas. We injected PAF in doses of 1, 4, 8, and 16 micrograms and normal saline as a control into the bullae of chinchillas. Vascular permeability was measured by the Evans blue vital dye technique. All the PAF-injected animals showed a significant increase in middle ear vascular permeability compared to the control group. This study demonstrated that PAF in the middle ear cavity contributes significantly to the development of OME by increasing the vascular permeability of the middle ear mucosa.

Efficacy of an aerosol surface spray against container-breeding Aedes.

The effectiveness of a residual insect surface spray against container-breeding mosquitoes was tested in Cairns, Australia. A formulation containing imiprothrin and cypermethrin (Mortein Plus Cockroach Lure 'n' Kill High Performance Surface Spray) was selected based on the label claim to "kill continuously for up to 6 months." A 1-sec spray was applied to the water and interior surface of partially flooded tires and terra-cotta pots. Treatments were paired with a control and replicated at 2 sites within 3 residential properties for a total of 6 replicates. All mosquito larvae were removed and counted weekly, and a representative sample was identified in the laboratory. Complete control of Aedes species was achieved for 4 and 5 months in all tires and pots, respectively.

Monte Carlo study of electron spectra produced in semi-infinite and finite water phantoms irradiated by photons of energies up to 2 MeV.

A Monte Carlo calculation for the initial electron energy spectra produced in semi-infinite and finite water phantoms irradiated by photons of energies up to 2 MeV and photons from 60Co are presented incorporating successive Compton scattering, photoelectric absorption and Auger effect. The backscattering of photons through the top of the phantom and transmission of photons out of the finite phantom through the sides also are considered. The results are compared with the earlier calculation for infinite water phantom of Turner et al. who assumed that all the photons remained in the water until they were absorbed. It is found that inclusion of backscattering in a semi-infinite phantom yields significantly different results from those of Turner et al. for the infinite phantom. It is also found that backscattering and transmission of the photons through the sides of the finite phantom further alter the initial energy spectra of electrons, as well as the average number of electrons per photon at different photon incident energies.