RNA interference-mediated tolerance to whitefly (Bemisia tabaci) in genetically engineered tomato

Plant Cell, Tissue and Organ Culture (PCTOC) - Whitefly (Bemisia tabaci) is a polyphagous insect that causes huge damage in several horticultural crops, including tomato, by sucking nutrients from...     

Taste perception in honeybees: just a taste of honey?

The advent of the genomic era has opened new doors to understand the fundamental organization of living organisms and has therefore promoted a fertile field of comparative research that intends to identify similarities and differences between related and unrelated species at the genomic level. One of the organisms whose genome has been recently decoded is that of the honeybee Apis mellifera, enabling a direct comparison with another well-studied insect, the fruit fly Drosophila melanogaster. It was reported that the honeybee has only ten gustatory receptors and thus a very poor taste perception compared to Drosophila, which presents 68 gustatory receptors, and the mosquito Anopheles gambiae, which presents 76 gustatory receptors. In this forum article, we discuss the implications of these findings taking into account previous and new discoveries on honeybee gustation based on behavioral and neurobiological studies by several authors and us. We conclude that the world of taste of a honeybee might not be as poor as proposed and that further studies should integrate molecular, neurobiological, behavioral and ecological approaches to better characterize taste perception in bees. Handling editor: Lars Chittka     

The ternary Rab27a-Myrip-Myosin VIIa complex regulates melanosome motility in the retinal pigment epithelium

The retinal pigment epithelium (RPE) contains melanosomes similar to those found in the skin melanocytes, which undergo dramatic light-dependent movements in fish and amphibians. In mammals, those movements are more subtle and appear to be regulated by the Rab27a GTPase and the unconventional myosin, Myosin VIIa (MyoVIIa). Here we address the hypothesis that a recently identified Rab27a- and MyoVIIa-interacting protein, Myrip, promotes the formation of a functional tripartite complex. In heterologous cultured cells, all three proteins co-immunoprecipitated following overexpression. Rab27a and Myrip localize to the peripheral membrane of RPE melanosomes as observed by immunofluorescence and immunoelectron microscopy. Melanosome dynamics were studied using live-cell imaging of mouse RPE primary cultures. Wild-type RPE melanosomes exhibited either stationary or slow movement interrupted by bursts of fast movement, with a peripheral directionality trend. Nocodazole treatment led to melanosome paralysis, suggesting that movement requires microtubule motors. Significant and similar alterations in melanosome dynamics were observed when any one of the three components of the complex was missing, as studied in ashen- (Rab27a defective) and shaker-1 (MyoVIIa mutant)-derived RPE cells, and in wild-type RPE cells transduced with adenovirus carrying specific sequences to knockdown Myrip expression. We observed a significant increase in the number of motile melanosomes, exhibiting more frequent and prolonged bursts of fast movement, and inversion of directionality. Similar alterations were observed upon cytochalasin D treatment, suggesting that the Rab27a-Myrip-MyoVIIa complex regulates tethering of melanosomes onto actin filaments, a process that ensures melanosome movement towards the cell periphery.     

Comparison of resistance to drought of three bean cultivars

The aim of the present work was to evaluate oxidative stress and plant antioxidant system of three contrasting bean (Phaseolus vulgaris L.) genotypes in the response to drought. Drought was imposed 14 d after emergence, by withholding water, until leaf relative water content reached 65 %. Water stress increased lipid peroxidation (LPO), membrane injury index, H₂O₂ and OH^⋅ production in leaves of stressed plants. Activities of the antioxidative enzymes superoxide dismutase (SOD) and ascorbate peroxidase (APOX) increased significantly under water stress in all the studied cultivars, while catalase (CAT) increased in cvs. Plovdiv 10 and Prelom, but decreased in cv. Dobrudjanski ran. Furthermore cv. Plovdiv 10 which had the highest APOX and CAT activities also showed the lowest increase in H₂O₂ and OH^⋅ production and LPO while cv. Dobrudjanski ran showed the lowest increases (and often the lowest values) in the antioxidant enzyme activities and the highest increases of H₂O₂ and OH^⋅ production, and LPO. On the basis of the data obtained we could specify cv. Plovdiv 10 and cv. Prelom as drought tolerant and cv. Dobrudjanski ran as a drought sensitive.     

Induction of apoptosis in HuH-7 cancer cells by monoterpene and β-carboline indole alkaloids isolated from the leaves of Tabernaemontana elegans

Three known (1–3) and a novel (4) monoterpene indole alkaloids have been isolated from the methanol extract of leaves of Tabernaemontana elegans and their structures were elucidated by a series of spectroscopic experiments, involving NMR, MS, UV, and IR techniques. The isolated monoterpene indole alkaloids along with previously described β-carbolines (5–7) from the same specimen were studied for their apoptosis induction activity in human hepatoma HuH-7 cells. Methodology for apoptosis induction studies included cell viability assays, nuclear morphology assessments, and general caspase-3-like activity assays. The monoterpene indole alkaloids, tabernaemontanine (1) and vobasine (3) showed the most promising apoptosis induction profile in HuH-7 cells.     

Drought effect on photosynthetic activity,osmolyte accumulation and membrane integrity of two <Emphasis Type="Italic">Cicer arietinum</Emphasis> genotypes

Drought was induced in chickpea (Cicer arietinum L.) genotypes (ChK 3226 and ILC 3279) differing in yield capacity. Water stress (S1, RWC around 55–50%; S2, RWC ≤ 40%) drastically reduced stomatal conductance (g _s) and net photosynthetic rate (P _N) in both genotypes. ILC 3279 showed greater photosynthetic capacity (A _max) decreases. Maximum PSII photochemical efficiency (F_v/F_m), photochemical quenching (q_P), total chlorophylls (Chls) and carotenoids (Cars) content showed stability in both genotypes under stress, but in S2 ILC 3279 presented an increase in basal fluorescence (F₀) and a greater reduction in estimation of quantum yield of linear electron transport (Φ_e) than ChK 3226. Membrane damage evaluated by electrolyte leakage occurred earlier and was greater in ILC 3279. It also presented a decrease of total fatty acids (TFA) along drought, while in ChK 3226 greater amounts of TFA were observed in S1. In rehydration, P _N of S1 plants completely recovered (ILC 3279) or remained slightly below control (ChK 3226). As regards S2 plants, ILC 3279 showed stronger P _N and g _s reductions than ChK 3226, despite both genotypes totally recovered A _max and chlorophyll (Chl) a fluorescence. ChK 3226 recovered more efficiently from membrane damage. Under control conditions, greater amounts of most of the studied soluble metabolites occurred in ChK 3226 plants. Malate and citrate decreased with water stress (S2) in both genotypes. Sucrose and pinitol (that had a higher concentration than sucrose in both genotypes) increased in ILC 3279 (S1 and S2), and decreased in ChK 3226 (S2). In ILC 3279 proline and asparagine followed similar patterns. Genotypes showed a similar shoot dry mass (DM) in control plants, but root DM was higher in ChK 3226. Drought reduced root and shoot DM in ChK 3226 already under S1, while in ILC 3279 root DM was unaffected by drought and shoot biomass decreased only in S2. Root/shoot ratio was always higher in ChK 3226 but tended to decrease under stress, while the opposite was observed in ILC 3279. No pods were obtained from control plants of both genotypes, or droughted ILC 3279 plants. ChK 3226 produced pods under S1 (higher yield) and S2. Under stress conditions, ChK 3226 was less affected in photosynthetic activity and membrane integrity, showing a better tolerance to drought. This agrees with the better yield of this genotype under water stress. Distinct strategies seem to underlie the different physiological responses of the two genotypes to water deficit. In spite of its significant solutes accumulation, ILC 3279 was more affected in photosynthetic activity and membrane integrity during water stress than ChK 3226, which showed better yield under drought. A relation could not be established between solutes accumulation of ILC 3279 and yield.     

Surprising negative association between IgG1 allotype disparity and anti-adalimumab formation: a cohort study

Introduction

The human monoclonal antibody adalimumab is known to induce an anti-globulin response in some adalimumab-treated patients. Antibodies against adalimumab (AAA) are associated with non-response to treatment. Immunoglobulins, such as adalimumab, carry allotypes which represent slight differences in the amino acid sequences of the constant chains of an IgG molecule. Immunoglobulins with particular IgG (Gm) allotypes are racially distributed and could be immunogenic for individuals who do not express these allotypes. Therefore, we investigated whether a mismatch in IgG allotypes between adalimumab and IgG in adalimumab-treated patients is associated with the development of AAA.

Methods

This cohort study consisted of 250 adalimumab-treated rheumatoid arthritis (RA) patients. IgG allotypes were determined for adalimumab and for all patients. Anti-idiotype antibodies against adalimumab were measured with a regular radio immunoassay (RIA), and a newly developed bridging enzyme linked immunosorbent assay (ELISA) was used to measure anti-allotype antibodies against adalimumab. The association between AAA and the G1m3 and the G1m17 allotypes was determined. For differences between groups we used the independent or paired samples t-test, Mann-Whitney test or Chi square/Fisher's exact test as appropriate. To investigate the influence of confounders on the presence or absence of AAA a multiple logistic regression-analysis was used.

Results

Adalimumab carries the G1m17 allotype. No anti-allotype antibodies against adalimumab were detected. Thirty-nine out of 249 patients had anti-idiotype antibodies against adalimumab (16%). IgG allotypes of RA patients were associated with the frequency of AAA: patients homozygous for G1m17 had the highest frequency of AAA (41%), patients homozygous for G1m3 the lowest frequency (10%), and heterozygous patients' AAA frequency was 14% (P = 0.0001).

Conclusions

An allotype mismatch between adalimumab and IgG in adalimumab-treated patients did not lead to a higher frequency of AAA. On the contrary, patients who carried the same IgG allotype as present on the adalimumab IgG molecule, had the highest frequency of anti-adalimumab antibodies compared to patients whose IgG allotype differed from adalimumab. This suggests that the allotype of adalimumab may not be highly immunogenic. Furthermore, patients carrying the G1m17-allotype might be more prone to antibody responses.     

Accumulation of CD1a-positive Langerhans cells and mast cells in actinic cheilitis

LCs and MCs are known to be directly influenced by UV radiation. This study investigated the presence of Langerhans cells (LCs) and mast cells (MCs) in actinic cheilitis (AC) exhibiting epithelial dysplasia (ED). Using immunohistochemistry for CD1a and mast cell tryptase, LCs and MCs density was assessed in 35 cases of AC with different degrees of ED. LCs were found in 32 cases of AC whereas MCs were found in all cases. There was an increase in LCs density irrespective of degree of ED when the cases were compared to normal lip mucosa (P = 0.04343). No statistical difference in LCs density was observed regarding the different degrees of dysplasia (P > 0.05). Significant difference in MCs density between mild and moderate dysplasia and normal lip mucosa was found (P < 0.05). No significant correlation between LCs and MCs was seen (P = 0.1258). Although no correlation could be established between LCs and MCs and the different degrees of ED; it is possible that the accumulation of LCs plays an immunostimulatory and protective role in the defense against progression of dysplasia. Further studies are necessary to determine the role of MCs in the development of AC.     

Exploiting the full power of temporal gene expression profiling through a new statistical test: Application to the analysis of muscular dystrophy data

Background  

The identification of biologically interesting genes in a temporal expression profiling dataset is challenging and complicated by high levels of experimental noise. Most statistical methods used in the literature do not fully exploit the temporal ordering in the dataset and are not suited to the case where temporal profiles are measured for a number of different biological conditions. We present a statistical test that makes explicit use of the temporal order in the data by fitting polynomial functions to the temporal profile of each gene and for each biological condition. A Hotelling T ²-statistic is derived to detect the genes for which the parameters of these polynomials are significantly different from each other.     

Nitric oxide is involved in the lesions of the peripheral autonomic neurons observed in the acute phase of experimental Trypanosoma cruzi infection

Our aim was to investigate the possible involvement of nitric oxide (NO) in peripheral denervation during the acute phase of murine experimental Trypanosoma cruzi infection. Wistar male rats were infected with the Y strain of T. cruzi. One group of animals was also treated with the NO synthase inhibitor N-nitro-l-arginine. A group of uninfected animals was the control. At the 18th day of infection the animals were sacrificed. Quantification of neurons in the colon and heart and tissue parasitism in the heart was performed. Serum concentration of nitrate was measured and a histochemical technique for assessing NADPH-diaphorase activity in the colon was also performed. The infected animals presented a statistically significant decrease in the number of peripheral neurons in the colon and heart and a 2-fold increase in serum NO(3) concentration compared with controls. The animals treated with N-nitro-l-arginine showed almost an absence of NO(3) concentration in the serum and did not show loss of neurons compared with controls. These treated animals displayed a 15-fold increase in tissue parasitism compared with nontreated infected animals. The NADPH-diaphorase activity was much more intense in the muscle layers of the colon of the infected animals than in those of the controls. Taken together, these data suggest that NO is involved in the peripheral denervation observed in the acute phase of experimental T. cruzi infection.