Dimethyl fumarate modulates neutrophil extracellular trap formation in a glutathione- and superoxide-dependent manner.

BACKGROUND: Neutrophil (polymorphonuclear) granulocytes (PMN) have been shown to contribute to the pathogenesis of psoriasis by releasing interleukin-17 and LL37-DNA complexes via neutrophil extracellular traps (NETs), webs of chromatin strands decorated with antimicrobial peptides, in psoriatic skin. Fumaderm® , a fumaric acid ester (FAE) formulation consisting of different FAE salts, has been successfully used to treat psoriasis for decades. Most recently, FAE treatment was reported to inhibit NET formation in murine epidermolysis bullosa acquisita. OBJECTIVES: To elucidate the effect of FAE treatment on human psoriasis and healthy donor NET formation. RESULTS: Among the compounds present in the FAE formulation, dimethyl fumarate (DMF) pretreatment of human psoriasis and healthy donor PMN resulted in a consistent inhibitory effect on NET formation in response to phorbol 12-myristate 13-acetate but not to platelet activating factor and ionomycin. This effect was l-glutathione (GSH) dependent and involved a decrease in reactive oxygen species (ROS) production, a key event in NET formation. In contrast, G-protein-coupled signalling and protein synthesis were not involved. Monomethyl fumarate (MMF) was found to slightly reduce ROS production without affecting NET formation. CONCLUSIONS: We report DMF as a potent, stimulus-specific, GSH- and ROS-dependent modulator of NET formation. Our results support the notion that modulation of NET formation contributes to the beneficial effects of FAEs in a variety of inflammatory conditions.

Altered density, composition and microanatomical distribution of infiltrating immune cells in cutaneous squamous cell carcinoma of organ transplant recipients.

BACKGROUND: The inflammatory tumour microenvironment is crucial for effective tumour control, and long-term immunosuppression has been identified as a major risk factor for skin carcinogenesis. In solid organ transplant recipients (OTRs) undergoing long-term pharmacological immunosuppression, an increased incidence of cutaneous squamous cell carcinoma (SCC) and more aggressive tumour growth compared with immunocompetent patients has been reported. OBJECTIVES: To determine the density and phenotype of immune cells infiltrating SCC and surrounding skin in OTRs, and to characterize the microanatomical distribution patterns in comparison with immunocompetent patients. METHODS: We analysed immune cell infiltrates within SCC and at defined regions of interest (ROIs) of tumour-surrounding skin in formalin-fixed paraffin-embedded tissue of 20 renal transplant patients and 18 carefully matched immunocompetent patients by high-resolution semiautomated microscopy on complete tissue sections stained for CD4, CD8, CD20 and CD68. RESULTS: The overall immune cell density of SCC arising in OTRs was significantly reduced compared with immunocompetent patients. Particularly CD4+ infiltrates at the directly invasive margin and tumour vicinity, intratumoral CD8+ T-cell densities and the overall density of CD20+ tumour-infiltrating B cells were significantly reduced in the tissue of OTRs. CONCLUSIONS: Immune cell infiltrates within SCC and at defined ROIs of tumour-surrounding skin in OTRs differ markedly in their composition and microanatomical distribution compared with tumours arising in immunocompetent patients. Our findings substantially broaden the understanding of how long-term systemic immunosuppression modulates the local inflammatory microenvironment in the skin and at the site of invasive SCC.

Some perspectives on the risks and benefits of biological control of invasive alien plants in the management of natural ecosystems.

Globally, invasions by alien plants are rapidly increasing in extent and severity, leading to large-scale ecosystem degradation. Weed biological control offers opportunities to arrest or even reverse these trends and, although it is not always effective or appropriate as a management strategy, this practice has an excellent record of safety and many notable successes over two centuries. In recent years, growing concerns about the potential for unintended, non-target damage by biological control agents, and fears about other unpredictable effects on ecosystems, have created an increasingly demanding risk-averse regulatory environment. This development may be counter-productive because it tends to overemphasize potential problems and ignores or underestimates the benefits of weed biological control; it offers no viable alternatives; and it overlooks the inherent risks of a decision not to use biological control. The restoration of badly degraded ecosystems to a former pristine condition is not a realistic objective, but the protection of un-invaded or partial restoration of invaded ecosystems can be achieved safely, at low cost and sustainably through the informed and responsible application of biological control. This practice should therefore be given due consideration when management of invasive alien plants is being planned. This discussion paper provides a perspective on the risks and benefits of classical weed biological control, and it is aimed at assisting environmental managers in their deliberations on whether or not to use this strategy in preference, or as a supplement to other alien invasive plant control practices.

Benefits of photosynthesis for insects in galls.

Insect-induced plant galls are predominantly reputed to act as strong carbon sinks, although many types of galls contain chlorophyll and have the potential to photosynthesize. We investigated whether the photosynthetic capacity of bud galls induced by a Pteromalid wasp, Trichilogaster acaciaelongifoliae, in Acacia longifolia subsidises carbon budgets or provides O(2) to the larvae while concurrently consuming CO(2) in the dense gall tissue, thereby maintaining (O(2)) and (CO(2)) within the range of larval tolerance. Low (O(2)) (<5 % v/v) were found within the internal tissues of galls, and these concentrations responded only marginally to light, suggesting that the photosynthetic activity within the gall is inconsequential in the provision of O(2) to the larvae. The metabolic response of larvae to reduced (O(2)) and elevated (CO(2)) indicated that larvae were tolerant of hypoxia/hypercarbia and also capable of reducing their respiratory rates to cope with hypercarbia. The low mortality of larvae in galls shaded with Al-foil for 20 days showed that photosynthesis was not vital for the survival of the larvae, although growth of shaded galls was substantially reduced. Gas exchange measurements confirmed that, while photosynthesis never fully compensated for the respiratory costs of galls, it contributed substantially to the maintenance and growth, especially of young galls, reducing their impact as carbon sinks on the host. We conclude that, although photosynthesis may contribute to O(2) provision, its main role is to reduce the dependence of the insect-induced gall on the host plant for photosynthates, thereby reducing intra-plant, inter-gall competition and enhancing the probability that each gall will reach maturity.

Autoantibodies in psoriasis as predictors for loss of response and anti-infliximab antibody induction.

BACKGROUND: Infliximab is successfully used to treat psoriasis and psoriatic arthritis. However, some patients lose therapeutic response after several cycles. Antibodies to infliximab (infliximab-Abs) are induced during treatment in a subgroup of patients and are thought to be associated with loss of response (LOR). Routine screening for infliximab-Abs is expensive and not regularly performed. A reliable and affordable method for identifying patients who are at risk for LOR to infliximab is desirable. OBJECTIVES: To analyse the development of antinuclear antibodies (ANA)/antidouble-stranded DNA antibodies (anti-dsDNA) over time in patients with psoriasis receiving infliximab. To analyse if there is an association between ANA titres/anti-dsDNA concentrations, infliximab-Ab status and LOR. METHODS: A retrospective data analysis of 29 patients with psoriasis receiving infliximab was carried out. ANA titres and anti-dsDNA concentrations were regularly monitored in these patients and sera were tested for infliximab-Abs by enzyme-linked immunosorbent assay. RESULTS: Median ANA titres increased from 1 : 80 [interquartile range (IQR) 0 to 1 : 320, n = 29] pretreatment, to 1 : 1280 (IQR 1 : 640 to 1 : 1920, n = 15) after infusion 10, and 1 : 1920 (IQR 1 : 1280 to 1 : 2560, n = 10) after infusion 20. Infliximab-Abs were found in 21% of patients. Infliximab-Ab-positive patients and patients with LOR had significantly higher pretreatment anti-dsDNA concentrations and higher pretreatment ANA titres than infliximab-Ab-negative and responsive patients, respectively. CONCLUSIONS: The results of this study suggest a role for autoantibodies in the identification of patients with psoriasis at higher risk of developing infliximab-Abs and of LOR under treatment with infliximab.

Hybridization between Dactylopius tomentosus (Hemiptera: Dactylopiidae) biotypes and its effects on host specificity.

Dactylopius tomentosus is composed of biotypes adapted to different Cylindropuntia species. One biotype is an important biological control agent of C. imbricata in South Africa while another has the potential for the control of C. fulgida var. fulgida. These two weed species occur in sympatry in some areas of South Africa, so the introduction of the second biotype could result in hybridization, which, in turn, could impact on the biological control programs through altered host specificity and fitness of the hybrids. To anticipate what might happen, reciprocal crosses were made between the two biotypes, and the biological performance of the resultant hybrids was compared with that of each parental lineage on C. imbricata and C. f. var. fulgida. The biotypes interbred freely and reciprocally in the laboratory. Comparisons of crawler and adult female traits showed differences in performance that were dependent on the origin of the maternal and paternal genomes. However, when all traits were combined into a 'fitness index', both hybrids clearly outperformed the parental lineages. The increase in fitness shown by the hybrids over their maternal lineage was greater on the alternative host of the maternal parent than on the natural host of the maternal parent. Therefore, in areas where the two cacti occur in sympatry, hybridization between the biotypes is not expected to be detrimental to the biological control of either weed.

Determination of biotypes of Dactylopius tomentosus (Hemiptera: Dactylopiidae) and insights into the taxonomic relationships of their hosts, Cylindropuntia spp.

Host specialization to form biotypes is common among phytophagous insects, and it has been hypothesised that biotypes of Dactylopius tomentosus L. (Hemiptera: Dactylopiidae) occur. D. tomentosus is an important biological control agent for Cylindropuntia cacti when they occur as weeds. Additionally, there is uncertainty surrounding the taxonomic status of some species of Cylindropuntia. This study aimed to confirm the existence of D. tomentosus biotypes and to assess whether host specialization can help to resolve this systematic uncertainty. For this study, the host specificity and performance of ten provenances of D. tomentosus collected from C. cholla, C. fulgida var. fulgida, C. imbricata, C. f. var. mamillata, C. rosea and C. tunicata and reared on C. cholla, C. fulgida var. fulgida, C. imbricata and C. rosea were investigated. Five life-history parameters were measured including: crawler development time and survival, female development time, and the weight and number of eggs produced by females. Results revealed significant variation in host specificity with provenances either thriving, surviving or dying on the different hosts, thus demonstrating the existence of biotypes. Also, host specificity was related to host species and not to the geographic locality from which either the host or provenance was sourced. These findings suggest that the characteristics of Cylindropuntia species may differ sufficiently, there by presenting different selection pressures that induce and sustain distinct biotypes of D. tomentosus. The observed host use patterns of the biotypes separated the plant species into two groups that accorded with known phylogenetic relationships among Cylindropuntia species, suggesting that biotypes can be used to elucidate their taxonomic relatedness. Besides advancing our knowledge of the ecology and evolution of D. tomentosus, these novel findings have important implications for the biological control of Cylindropuntia species.

The biology of Dactylopius tomentosus (Hemiptera: Dactylopiidae).

Dactylopius tomentosus (Lamarck) (Hemiptera: Dactylopiidae) is a cochineal insect whose host range is restricted to Cylindropuntia species (Caryophyllales: Cactaceae). This insect has been utilized successfully for biological control of Cylindropuntia imbricata (Haw.) F.M. Knuth in Australia and South Africa. Despite this, its biology has not been studied previously, probably due to the widely held belief that the biology of all Dactylopius species is similar. This study investigated the life cycle and the morphological and reproductive characteristics of D. tomentosus. Results revealed some unique characteristics of D. tomentosus: (i) eggs undergo a much longer incubation period, an average of 17 days compared to <1 day in its congeners; (ii) eggs are laid singly but are retained as an egg mass secured in a mesh of waxy threads attached to the female; (iii) the developmental times of males and females are longer compared to other Dactylopius spp. due to a longer egg incubation period; (iv) D. tomentosus does not undergo parthenogenesis; (v) D. tomentosus is smaller in size than its congeners; and (vi) male mating capacity and reproductive potential were both high and variable between males. There was a significant, strong, positive relationship (r = 0.93) between female mass and fecundity, whereas the relationship between the number of females mated per male that became gravid and their fecundity was negative (r = -0.68). Besides contributing to our knowledge of this economically important species, the finding of unique characteristics of D. tomentosus biology underlines the need to study each species in this genus.

Distinguishing suitable biotypes of Dactylopius tomentosus (Hemiptera: Dactylopiidae) for biological control of Cylindropuntia fulgida var. fulgida (Caryophyllales: Cactaceae) in South Africa.

Cylindropuntia fulgida (Engelmann) F.M. Knuth var. fulgida (Engelmann) F.M. Knuth (Cff) (Caryophyllales: Cactaceae) is native to Mexico and Arizona and was introduced into South Africa for ornamental purposes. It subsequently became highly invasive, necessitating control. The cochineal insect, Dactylopius tomentosus (Lamarck) (Hemiptera: Dactylopiidae), was selected as a potential biological control agent based on its restricted host range among Cylindropuntia species and previous success in controlling C. imbricata (DC.) F. Knuth (Ci). Eight D. tomentosus provenances (Cholla, Cholla E, Fulgida, Mamillata, Imbricata, Tunicata U, Tunicata V and Rosea) from Cylindropuntia species in their native ranges were reared on Cff, whilst Cholla and Imbricata were also reared on Ci. Large differences were found in the development and survival of crawlers, and in the reproductive capacity of females. Three subjective categories of provenance interaction with host plants were identified based on a fitness index (FI) calculated from data relating to crawler survival, female development time and fecundity: (i) thriving (FI > or = 1) - insects had shorter developmental times, high crawler survival and highly fecund females (Cholla); (ii) surviving (FI<1 but >0) - insects had extended development times, low crawler survival and low fecundity (Imbricata, Fulgida and Mamillata); and (iii) dying (FI = 0) - insects died before or at the second instar (Rosea, Tunicata U and Tunicata V). Cholla, therefore, is highly suitable for biological control of Cff in South Africa. In addition, Cholla thrived on Cff but only survived on Ci whilst, in contrast, Imbricata thrived on Ci but only survived on Cff. This differential ability of provenances to thrive or survive on different host plants demonstrated that host adapted biotypes of D. tomentosus exist; therefore, biotypes should be taken into account when considering this species as a biological control agent of cactus weeds.

Activation of the redox-regulated molecular chaperone Hsp33--a two-step mechanism.

BACKGROUND: Hsp33 is a novel redox-regulated molecular chaperone. Hsp33 is present in the reducing environment of the cytosol and is, under normal conditions, inactive. The four highly conserved cysteines found in Hsp33 constitute a novel zinc binding motif. Upon exposure to oxidative stress, Hsp33's chaperone activity is turned on. This activation process is initiated by the formation of two intramolecular disulfide bonds. Recently, the 2.2 A crystal structure of Hsp33 has been solved, revealing that Hsp33 is present as a dimer in the structure (Vijayalakshmi et al., this issue, 367-375 [1]). RESULTS: We show here that oxidized, highly active Hsp33 is a dimer in solution. In contrast, reduced and inactive Hsp33 is monomeric. The incubation of reduced Hsp33 in H(2)O(2) leads to the simultaneous formation of two intramolecular disulfide bonds and the concomitant release of zinc. This concentration-independent step is followed by a concentration-dependent association reaction. The dimerization of Hsp33 requires highly temperature-sensitive structural rearrangements. This allows Hsp33's activation process to be greatly accelerated at heat shock temperatures. CONCLUSIONS: The regulation of Hsp33's chaperone function is highly sophisticated. On a transcriptional level, Hsp33 is under heat shock control. This increases the concentration of Hsp33 under heat and oxidative stress, a process that favors dimerization, a critical step in Hsp33's activation reaction. On a posttranslational level, Hsp33 is redox regulated. Dimerization of disulfide-bonded Hsp33 monomers leads to the formation of two extended, putative substrate binding sites. These sites might explain Hsp33's high and promiscuous affinity for unstructured protein folding intermediates.

Biocontrol of a perennial legume, Sesbania punicea, using a florivorous weevil, Trichapion lativentre: weed population dynamics with a scarcity of seeds.

The establishment in South Africa of a florivorous, apionid weevil, Trichapion lativentre, on Sesbania punicea, a leguminous weed of South American origin, has reduced seed production of the plants by >98%. Surveys of the age structure and density of plants in infestations of S. punicea throughout South Africa have shown that the rate of recruitment of seedlings has drastically declined within a few years in many areas, due to the weevils. However, there has unexpectedly not been a corresponding decline in the density of mature plants in extant infestations of S. punicea. In spite of this, T. lativentre has curtailed the rate of spread of the weed into uninvaded habitats and has impeded reinvasion into areas cleared of infestations by mechanical means or by another complimentary biocontrol agent.

The population dynamics of an introduced tree, Sesbania punicea, in South Africa, in response to long-term damage caused by different combinations of three species of biological control agents.

This paper contributes to the relatively sparse literature on the effects of insect herbivory on the population dynamics of plants and is probably unique in that it reports the long-term effects of combinations of three insect herbivore species on the population densities of a moderately long-lived tree species. The tree is Sesbania punicea, a leguminous perennial from South America that has been the target of a biological control programme in South Africa for almost 20 years. Sixteen infestations of the weed have been monitored for periods of up to 10 years to determine changes in the density of the mature, reproductive plants under the influence of different combinations of three biological control agents (i.e. with one, two or three of the agent species present in the weed infestation). The three biological control agents, all weevil species, include Trichapion lativentre, which primarily destroys the flower-buds, Rhyssomatus marginatus, which destroys the developing seeds, and Neodiplogrammus quadrivittatus, whose larvae bore into the trunk and stems of the plants. While T. lativentre occurs throughout the range of the weed in South Africa, the other two species are less mobile, more recent introductions and are largely confined to the vicinity of selected release sites. There has been a significant decline in the density of mature S. punicea in areas where two or more of the agents are established. The decline of the weed has been most evident where N. quadrivittatus is active and particularly so where both of the other two weevil species are also present.

[Magnetoneurographic registration of evoked summation action fields over lumbar vertebrae following transcutaneous tibial nerve stimulation]. / Magnetoneurographische Darstellung evozierter Summenaktionsfelder über der lumbalen Wirbelsäule nach transkutaner N. Tibialis Stimulation.

Goal of this study was the development of a protocol for the registration of evoked magnetic fields over the lumbar spine using off-the-shelf equipment. Three subjects in a sitting position with their torso bent slightly forward were stimulated at the tibial nerve with a commercially available stimulator. Neuromagnetic fields were registered over a circular, 800 cm2 area of the lumbosacral spine using a 61-channel 4D-Neuroimaging biomagnetometer. After appropriate signal processing, dipolar magnetic fields with a field strength 5-17 fT peak-to-peak amplitude were detected in three out of four registrations. Location and orientation of these fields concurred with the expected evoked compound action currents along the course of the nerve fibers.