I'm Infected, Eat Me! Innate Immunity Mediated by Live, Infected Cells Signaling To Be Phagocytosed.

Innate immunity against pathogens is known to be mediated by barriers to pathogen invasion, activation of complement, recruitment of immune cells, immune cell phagocytosis of pathogens, death of infected cells, and activation of the adaptive immunity via antigen presentation. Here, we propose and review evidence for a novel mode of innate immunity whereby live, infected host cells induce phagocytes to phagocytose the infected cell, thereby potentially reducing infection. We discuss evidence that host cells, infected by virus, bacteria, or other intracellular pathogens (i) release nucleotides and chemokines as find-me signals, (ii) expose on their surface phosphatidylserine and calreticulin as eat-me signals, (iii) release and bind opsonins to induce phagocytosis, and (iv) downregulate don't-eat-me signals CD47, major histocompatibility complex class I (MHC1), and sialic acid. As long as the pathogens of the host cell are destroyed within the phagocyte, then infection can be curtailed; if antigens from the pathogens are cross-presented by the phagocyte, then an adaptive response would also be induced. Phagocytosis of live infected cells may thereby mediate innate immunity.

Automated 3D bioassembly of micro-tissues for biofabrication of hybrid tissue engineered constructs.

Bottom-up biofabrication approaches combining micro-tissue fabrication techniques with extrusion-based 3D printing of thermoplastic polymer scaffolds are emerging strategies in tissue engineering. These biofabrication strategies support native self-assembly mechanisms observed in developmental stages of tissue or organoid growth as well as promoting cell-cell interactions and cell differentiation capacity. Few technologies have been developed to automate the precise assembly of micro-tissues or tissue modules into structural scaffolds. We describe an automated 3D bioassembly platform capable of fabricating simple hybrid constructs via a two-step bottom-up bioassembly strategy, as well as complex hybrid hierarchical constructs via a multistep bottom-up bioassembly strategy. The bioassembly system consisted of a fluidic-based singularisation and injection module incorporated into a commercial 3D bioprinter. The singularisation module delivers individual micro-tissues to an injection module, for insertion into precise locations within a 3D plotted scaffold. To demonstrate applicability for cartilage tissue engineering, human chondrocytes were isolated and micro-tissues of 1 mm diameter were generated utilising a high throughput 96-well plate format. Micro-tissues were singularised with an efficiency of 96.0 ± 5.1%. There was no significant difference in size, shape or viability of micro-tissues before and after automated singularisation and injection. A layer-by-layer approach or aforementioned bottom-up bioassembly strategy was employed to fabricate a bilayered construct by alternatively 3D plotting a thermoplastic (PEGT/PBT) polymer scaffold and inserting pre-differentiated chondrogenic micro-tissues or cell-laden gelatin-based (GelMA) hydrogel micro-spheres, both formed via high-throughput fabrication techniques. No significant difference in viability between the construct assembled utilising the automated bioassembly system and manually assembled construct was observed. Bioassembly of pre-differentiated micro-tissues as well as chondrocyte-laden hydrogel micro-spheres demonstrated the flexibility of the platform while supporting tissue fusion, long-term cell viability, and deposition of cartilage-specific extracellular matrix proteins. This technology provides an automated and scalable pathway for bioassembly of both simple and complex 3D tissue constructs of clinically relevant shape and size, with demonstrated capability to facilitate direct spatial organisation and hierarchical 3D assembly of micro-tissue modules, ranging from biomaterial free cell pellets to cell-laden hydrogel formulations.

Phagoptosis - Cell Death By Phagocytosis - Plays Central Roles in Physiology, Host Defense and Pathology.

Cell death by phagocytosis - termed 'phagoptosis' for short - is a form of cell death caused by the cell being phagocytosed i.e. recognised, engulfed and digested by another cell. Phagocytes eat cells that: i) expose 'eat-me' signals, ii) lose 'don't-eat-me' signals, and/or iii) bind opsonins. Live cells may express such signals as a result of cell stress, damage, activation or senescence, which can result in phagoptosis. Phagoptosis may be the most abundant form of cell death physiologically as it mediates erythrocyte turnover. It also regulates: reproduction by phagocytosis of sperm, development by removal stem cells and excess cells, and immunity by removal of activated neutrophils and T cells. Phagoptosis mediates the recognition of non-self and host defence against pathogens and cancer cells. However, in inflammatory conditions, excessive phagoptosis may kill our cells, leading to conditions such as hemophagy and neuronal loss.

Vascular crossing patterns in patients with systemic arterial hypertension.

AIM: To identify the relationship of retinal arteries in a population with systemic arterial hypertension. METHODS: High resolution, dilated, digitised, fundus photographs of consecutive patients with a history of hypertension requiring pharmacologic therapy seen on the Wills Eye Hospital Retina Service were analysed. Included were photographs of the temporal retinal vascular arcades in which media clarity permitted good visualisation of third-order retinal vascular bifurcations. Each first- and second-order arteriovenous (AV) crossing was then examined to identify anatomic patterns at the sites where veins and arteries crossed. Eyes in patients without a history of hypertension were used as controls. RESULTS: Among the 71 patients (134 eyes), there were 430 first-order and second-order AV crossings, in which AV nicking was present at 126 sites. A retinal artery was located anterior to the retinal vein in 122 of the 126 sites (96.8%) at which AV nicking was noted, while nicking associated with the vein located anteriorly to the artery occurred in only 4 of 126 (3.2%) of AV crossings (p<0.001). An anatomical pattern of venous arching, or cascading of a retinal vein over a retinal artery, was noted predominantly when the vein was positioned anterior to the artery in both subjects and controls. Among the 43 venous arching sites in the study group, 41 (95.3%) demonstrated the retinal vein anterior to retinal artery (p<0.001). CONCLUSIONS: In patients with systemic arterial hypertension and hypertensive retinopathy, AV nicking of the retinal vein at the site of AV crossing is seen predominantly when the retinal artery lies anterior to the vein, but generally not when the vein lies anterior to the artery. The clinician should realise that when a retinal vein lies anterior to a retinal artery, the absence of AV nicking does not rule out more severe, chronic, retinopathic changes than observed with retinal arterial straightening only.

Spin density projection-assisted R2 magnetic resonance imaging of the liver in the management of body iron stores in patients receiving multiple red blood cell transfusions: an audit and retrospective study in South Australia.

AIM: To assess the impact of non-invasive monitoring of liver iron concentration (LIC) on management of body iron stores in patients receiving multiple blood transfusions. METHOD: A retrospective audit was conducted on clinical data from 40 consecutive subjects with haemolytic anaemias or ineffective haematopoiesis who had been monitored non-invasively for LIC over a period of at least 1 year. LIC was measured with spin density projection-assisted proton transverse relaxation rate-magnetic resonance imaging. RESULTS: Nineteen clinical decisions were explicitly documented in the case notes as being based on LIC results. Decisions comprised initiation of chelation therapy, increasing chelator dose, decreasing chelator dose and change of mode of delivery of deferioxamine from subcutaneous to intravenous. The geometrical mean LIC for the cohort dropped significantly (P= 0.008) from 6.8 mg Fe/g dry tissue at initial measurement to 4.8 mg Fe/g dry tissue at final measurement. The proportion of subjects with LIC in the range associated with greatly increased risk of cardiac disease and death (>15 mg Fe/g dry tissue) dropped significantly (P= 0.01) from 14 of 40 subjects at initial measurement to 5 of 40 subjects at final measurement. No significant changes in the geometrical mean of serum ferritin or the proportion of subjects with serum ferritin above 2500 or 1500 µg/L were observed. CONCLUSIONS: The data are consistent with previous observations that introduction of non-invasive monitoring of LIC can contribute to a decreased body iron burden through improved clinical decision making and improved feedback to patients and hence improved adherence to chelation therapy.

The influence of Lasius neoniger (Hymenoptera: Formicidae) on population growth and biomass of Aphis glycines (Hemiptera: Aphididae) in soybeans.

In the United States, the soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), are often tended by the aphid-tending ant, Lasius neoniger Emery (Hymenoptera: Formicidae). In this study, we examined the effects of tending by ants on the density and biomass of soybean aphids on soybeans in Kentucky. We performed cage studies that limited access by ants and/or natural enemies. We used a split-plot design with natural enemy access as the main plot and ant attendance as the sub plot. We found that natural enemy access negatively affected aphid population density in the presence of tending ants, seen as a three- to four-fold increase in aphid density when natural enemies were excluded. In addition, we found that ant tending positively affected aphid biomass, both when natural enemies were given access to aphids or when natural enemies were excluded, seen by a two-fold increase in aphid biomass when ants tended aphids, both in the presence or absence of natural enemies. Biomass accumulation is seen as an important measurement for assessing aphid performance, and we argue that aphid-tending by ants can have an influence on natural field populations of soybean aphids. Agronomic practices that affect ant abundance in soybeans may influence the performance and hence pest outbreaks for this economically important pest.

The NO donor DETA-NONOate reversibly activates an inward current in neurones and is not mediated by the released nitric oxide.

BACKGROUND AND PURPOSE: It has been previously shown that high levels of nitric oxide (NO), from NO donors, kill neurones, but the mechanisms are unclear. EXPERIMENTAL APPROACH: The effects of NO donors on the electrical properties of rat cultured cerebellar granule cells (CGC neurones) were investigated using the whole-cell patch-clamp technique. KEY RESULTS: The NO donor (Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate (DETA-NONOate or NOC-18) caused a rapid, persistent, but fully reversible inward current that was associated with an increase in baseline noise and was concentration dependent (100 microM-10 mM). The response to 3 mM DETA-NONOate was completely inhibited by 1 mM gadolinium, but not by NO scavengers (1 mM haemoglobin or 1 mM PTIO) or glutamate receptor antagonists (10 microM MK-801 or 60 microM CNQX). Application of decomposed 3 mM DETA-NONOate or 3 mM nitrite had no effect. In contrast, the NO donor S-nitrosoglutathione (GSNO) caused a rapid, persistent, but fully reversible outward current that was also concentration dependent (1-10 mM). The 3 mM GSNO response was unaltered by NO scavengers, glutamate antagonists or gadolinium, but was mimicked by decomposed 3 mM GSNO and 3 mM oxidized glutathione. CONCLUSIONS AND IMPLICATIONS: These results suggest that DETA-NONOate directly activates cation-selective channels, causing an inward current in CGCs. In contrast, GSNO causes an outward current in these cells. Some of the effects of these NO donors are independent of NO, and thus caution is required in interpreting results when using high concentrations of these compounds.

New state records for Lutzomyia shannoni and Lutzomyia vexator.

Two species of phlebotomine sand flies, Lutzomyia shannoni (Dyar) and Lutzomyia vexator (Coquillett), are reported for the first time from Kentucky and Ohio. L. vexator also is reported for the first time from Tennessee. These insects were found in a northeasterly band extending from southwestern Kentucky to southwestern Ohio. Both species were consistently captured from mid-July through September in 2006 and 2007 by using CO2-baited Center for Disease Control light traps. Weekly sampling revealed that these flies are more abundant in the southern part of this band than in the northern part, but increasing densities throughout this new range indicate that the flies are currently expanding their range. Although both species have been reported further north along the Atlantic coast, and L. vexator along the Pacific coast, neither of them had been reported this far north along the Mississippi Valley. Previous reports established L. shannoni as far north as west central Tennessee and L. vexator in a similar spatial pattern in the eastern part of its range, extending as far north as northern Alabama. Whether the new records reported herein represent a northerly expansion of the geographic range of these species or are reflective of sampling changes is inconclusive. However, the former scenario could presage an increased prevalence of the diseases associated with this group of insects.

The power of hope: being a doctor is more than relying solely on the numbers.

AIMS: To ascertain whether a physician's positive or negative attitude significantly impacts the quality of life of ophthalmic patients. METHODS: A standardised, validated, time trade-off, utility instrument was administered to consecutive vitreoretinal patients by interview to assess the quality of life associated with their current ocular health state (baseline scenario). Each was then given a scenario for the exact same health state with the same long-term prognosis in which their doctor emphasised the possible negative consequences (bad-news scenario) and one for the same health state in which their doctor emphasised the positive consequences (good-news scenario). RESULTS: Among the 247 patients enrolled were 140 women (57%) and 107 men (43%) with a mean age of 66 years and a mean educational level of 13.8 years after kindergarten. The mean baseline utility for 247 patients was 0.87 (SD = 0.19; 95% CI 0.84 to 0.89). The mean bad-news scenario utility was 0.80 (SD = 0.22, 95% CI 0.78 to 0.83), a 70% diminution in quality of life compared with the mean baseline utility (p = 0.0009). The mean good-news scenario utility was 0.89 (SD = 0.18, 95% CI 0.86 to 0.91), an insignificant difference compared with the mean baseline utility (p = 0.26). CONCLUSION: Ocular patients had a considerably poorer quality of life when their physician emphasised the possible negative consequences associated with their eye disease(s), as opposed to a more positive approach. While at times necessary, a negative emphasis approach can theoretically result in a considerable loss of life's value.

The effect of comorbidities upon ocular and systemic health-related quality of life.

AIM: The purpose of the study was to assess whether, and to what degree, comorbidities affect patient quality of life. METHODS: A cross-sectional, quality-of-life study of 170 consecutive vitreoretinal patients compared the utility associated with a participant's primary (most incapacitating) disease and the utility associated with a grouping of all of the participants' diseases. The ocular diseases present included diabetic retinopathy (44%), macular degeneration (30%), lattice degeneration/retinal tear (14%), retinal vascular obstruction (5%), uveitis, macular oedema, macular pucker (5%) and others (2%). Participants underwent interviewer-administered, time trade-off utility questions for each disease, then for a compilation of all diseases. Their primary disease was defined by the lowest utility reported for a single disease, while other health conditions were considered comorbidities. A two-tailed, paired t test was used to compare the means of the primary disease utilities and compilation utilities. The study was powered to have a 90% chance of detecting an 8% difference in mean utility between the two utility groups RESULTS: The mean lowest utility for the most disabling single health condition (primary disease) was 0.82 (SD 0.22; 95% CI 0.79 to 0.85. The mean utility for the grouping together of all diseases was 0.80 (SD 0.24, 95% CI 0.76 to 0.84). No significant difference was found between the mean utilities of the two groups (p = 0.56). CONCLUSIONS: The overall health-related quality of life of a patient in an ophthalmic population with serious diseases appears to be primarily determined by the single disease that most adversely affects the individual's quality of life. This conclusion has significant implications in clinical care and when considering the use of comorbidities in cost-utility analyses.

Mechanisms of inflammatory neurodegeneration: iNOS and NADPH oxidase.

Inflammation contributes to a wide variety of brain pathologies, apparently via glia killing neurons. A number of mechanisms by which inflammatory-activated microglia and astrocytes kill neurons have been identified in culture. These include iNOS (inducible nitric oxide synthase), which is expressed in glia only during inflammation, and PHOX (phagocytic NADPH oxidase) found in microglia and acutely activated by inflammation. High levels of iNOS expression in glia cause (i) NO (nitric oxide) inhibition of neuronal respiration, resulting in neuronal depolarization and glutamate release, followed by excitotoxicity, and (ii) glutamate release from astrocytes via calcium-dependent vesicular release. Hypoxia strongly synergizes with iNOS expression to induce neuronal death via mechanism (i), because NO inhibits cytochrome oxidase in competition with oxygen. Activation of PHOX (by cytokines, beta-amyloid, prion protein, ATP or arachidonate) causes microglial proliferation and inflammatory activation; thus PHOX is a key regulator of inflammation. Activation of PHOX alone causes no death, but when combined with expressed iNOS results in extensive neuronal death via peroxynitrite production.

Neurodegeneration in models of Gram-positive bacterial infections of the central nervous system.

Gram-positive bacterial infections of the central nervous system, such as meningitis, induce an extensive inflammatory response, which in turn may damage neurons. LTA (lipoteichoic acid) is a component of the Gram-positive bacterial cell wall that induces glial inflammatory activation in vitro and in vivo. It does so by binding to Toll-like receptor-2 on microglia and astrocytes, rapidly activating ERK1/2 (extracellular-signal-regulated kinase 1/2) and p38 MAPKs (mitogen-activated protein kinases), causing NF-kappaB (nuclear factor kappaB) activation and leading to the production of pro-inflammatory cytokines and expression of inducible nitric oxide synthase (in synergy with muramyl dipeptide). LTA-activated microglia kill co-cultured neurons apparently via nitric oxide, superoxide and peroxynitrite, which may induce apoptosis of neurons that are then phagocytosed by microglia.

Efficacy of two pyrethroid insecticides applied as barrier treatments for managing mosquito (Diptera: Culicidae) populations in suburban residential properties.

Increased threat of mosquito-borne disease coupled with decreased tolerance of nuisance mosquitoes has opened a market for pest management professionals to offer mosquito control services for homeowners. A pest management professional applied bifenthrin (0.08%) and lambda-cyhalothrin (0.1%) at their maximum label concentrations as barrier treatments. We tested treatments residual efficacy in reducing adult mosquito populations and compared these chemicals against a water control at 24 residential properties (eight replications by three treatments). Mosquito populations were measured on each property by using five methods: CO2-baited Centers for Disease Control (CDC) light traps (without a light), human landing rates, CDC gravid traps, ovitraps, and sweep nets. Populations were monitored weekly for 2 wk before treatment and 8 wk posttreatment. Additionally, to confirm residual efficacy of each insecticide, a randomly treated leaf underwent a no-choice bioassay with laboratory-reared Aedes albopictus (Skuse). Trap collections were dominantly Aedes albopictus and Culex pipiens L. Both insecticidal treatments significantly reduced Aedes spp. lambda-Cyhalothrin- and bifenthrin-treated sites had 89.5 and 85.1% fewer Ae. albopictus bites than the untreated control, respectively. Ae. albopictus bioassay results showed significant residual efficacy for both insecticides up to 6 wk posttreatment. There were no significant differences between properties treated with the two insecticides. In contrast, Culex spp. were not reduced by either insecticidal treatment. Our study indicated that barrier sprays applied to low-lying vegetation do not properly target adult daytime resting sites for Culex mosquitoes but that they can reduce Aedes mosquitoes. Perhaps by treating upper tree canopies Culex spp. abundance may be reduced.

Interactions between nitric oxide, oxygen, reactive oxygen species and reactive nitrogen species.

ROS (reactive oxygen species) and RNS (reactive nitrogen species) are central to the innate immunity that protects us from infection, but also contribute to degenerative diseases and possibly aging. However, ROS and RNS are increasingly recognized to contribute to physiological signalling. This review briefly describes the main interactions between ROS and RNS and shows how their origins, chemistry, metabolism and biological actions are intimately linked.

European corn borer (Lepidoptera: Crambidae) effects on qualitative traits of high oil content corn grown in Kentucky.

The European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), is one of the most important pests of corn, Zea mays L., because it consistently causes high loss of yield. A study was conducted in 2000-2002 at field sites in central and western Kentucky to investigate whether infestation by O. nubilalis differentially affects the production of high-oil corn compared with traditional field corn. Statistical differences in grain weight and percentage of oil content between the five infestation levels were significant at both locations and for all years. Average grain yield was reduced by 0.40% and average oil concentration by 0.011% for each 1% of damaged plants, and there was a strong correlation (0.76) between leaf damage ratings (i.e., Guthrie scale) and yield reduction. In general, corn planted at the early planting date tended to have a higher yield (grain weight) and oil content.

Neuropsychological and information processing performance and its relationship to white matter changes following moderate and severe traumatic brain injury: a preliminary study.

Reductions in information processing speed have frequently been reported following moderate and severe traumatic brain injuries (TBIs), consistent with the effects of diffuse white matter damage. Although the corpus callosum (CC) is a common site for diffuse damage following TBI, the effects of this damage on information processing speed have not been adequately examined. This study assessed a TBI group and a matched control group on tests of attention, memory, fluency, and set shifting ability, together with reaction time (RT) tasks requiring the inter- and intrahemispheric processing of visual and tactile information. The RT tasks were designed to target the cognitive functions that are likely to be affected by diffuse white matter damage, including damage to the CC. The TBI group demonstrated deficits in verbal and visual fluency and verbal memory. They were also slower on the visual and tactile RT tasks, were more affected by task complexity, and slower on RT tasks requiring the interhemispheric transfer of information. In fact, one of the interhemispheric tactile RT tasks proved to be the most discriminating of all the cognitive and RT measures. MRIs completed on a subset of TBI participants indicated that the mean CC measurements were 5% to 19% smaller than a normative control group, with the most atrophied areas being the isthmus and anterior midbody. Although white matter atrophy was moderately related to visual and tactile RT performance, and total hippocampal volume related to memory performance, CC area was not related to many of the tasks that were designed to tap interhemispheric processing. None of the standard cognitive tests correlated with outcome in the TBI group, but 1 of the tactile RT measures was significantly related to 2 measures of outcome.

Nitric oxide, hypoxia and brain inflammation.

NO (nitric oxide) acutely and potently inhibits mitochondrial cytochrome oxidase in competition with oxygen, thereby raising the apparent K(M) for oxygen of mitochondria and neurons into the physiological or pathological range. We find that NO from an NO donor or glial inducible NOS (nitric oxide synthase) highly sensitizes neurons to hypoxia-induced death, probably via the NO-oxygen competition at cytochrome oxidase. Thus the NO from neuronal NOS during excitotoxicity or the NO from inducible NOS during inflammation may sensitize the brain to hypoxic/ischaemic damage.

Bionomics of Scymnus (Pullus) louisianae J. Chapin (Coleoptera: Coccinellidae) as a predator of the soybean aphid, Aphis glycines Matsumura (Homoptera: Aphididae).

A field population of Scymnus louisianae Chapin (Coccinellidae) was found attacking soybean aphids, Apis glycines Matsumura (Aphidae), a pest recently introduced into Kentucky. This coccinellid had not previously been found in Kentucky. A greenhouse population of S. louisianae was established and its predation on A. glycines studied under laboratory conditions. Total time to develop from egg to adult was about 20 d. About 70% of immatures survived to adulthood and they consumed approximately 100 aphid nymphs per beetle larva during the beetle's four larval instars. Adults lived for an average of 47 d (mated males) and 63 d (mated females) and, during their total adult lifetime, mated males consumed an average of 665 nymphs and mated females consumed 1261 nymphs. All developmental times and predation rates were comparable to those reported for other aphidophagous Scymnus spp. which, in conjunction with reports that Scymnus spp. are effective predators of cotton aphids, Aphis gossypii Glover, suggests that S. louisianae is a potentially important predator of A. glycines in the southern United States.