Behavioral and electrophysiological responses of the parasitic wasp Psyttalia concolor (Szépligeti) (Hymenoptera: Braconidae) to Ceratitis capitata-induced fruit volatiles

Psyttalia concolor (Szépligeti) is a koinobiont larval-pupal endoparasitoid of many Tephritidae of great economic importance, such as the medfly, Ceratitis capitata (Wiedemann). In several species of parasitoids it has been demonstrated that the mated females are strongly attracted by specific volatiles from insect-damaged plants. Yet the role of olfactory cues deriving from C. capitata-infested fruits on the female’s decision during the P. concolor host location was poorly investigated. In the present study, the responses of P. concolor females to either healthy or C. capitata-infested fruits was studied through behavioral assays. Volatiles emitted by healthy and infested fruits were SPME-sampled and analyzed by gas chromatography–mass spectrometry (GC–MS). The attractiveness of the identified volatiles was assessed and their electrophysiological activity was analyzed through gas-chromatography coupled with electroantennography (GC-EAD). P. concolor preferred infested peaches and apples over healthy ones, either when visual and olfactory or only olfactory cues were given. Nine compounds were found as exclusive of infested peaches, with respect to healthy ones, and seven of them evoked electrophysiological responses. In apples only quantitative changes in volatile emissions were observed after the medfly infestation. The emissions of 1-butyl butylate, 1-hexyl acetate and 1-butyl esanoate increased in infested apples, whereas 1-hexyl (E)-2-methyl butenoate decreased significantly. Among apple volatiles, 1-butyl butylate, 2-methyl-1-butyl acetate, 1-hexyl acetate, 2-methyl-1-butyl 2-methylbutanoate, 1-butyl hexanoate and 1-hexyl (E)-2-methyl butenoate elicited responses in female antennae. Synthetic blends reproducing the odors emitted by infested peaches and apples elicited strong attraction towards P. concolor females. For both fruits, the blend attractiveness was mainly due to some specific electrophysiological active chemicals: ethyl octanoate, decanal and 4-decanolide for peach, and 1-butyl butylate and 1-butyl hexanoate for apple. The responses induced by the identified fruit volatiles to P. concolor females allow us to suppose that they play a role as short-range attractants during host location.     

Effect of environmental conditions on decomposition in eight woody species of a dry tropical forest

Litter decay is a significant part of carbon budget. Due to strong environmental control, the changes in the environment may drastically influence the litter decay rates. Litter decomposition of eight dry tropical woody species, viz. Shorea robusta, Buchanania lanzan, Diospyros melanoxylon, Lagerstroemia parviflora, Lannea coromandelica, Terminalia tomentosa, Holarrhena antidysenterica and Lantana camara was studied to document the effect of intra-annual changes in the environment. Litter decomposition was monitored at monthly intervals at five sites using litter bag technique over an annual cycle in a dry tropical deciduous forest of Vindhyan highland, India. Weight loss differed among species and through months, and ranged from 15.38% in L. camara at Kotwa site in January to 30.72% in T. tomentosa at Hathinala site in August. Peak weight loss occurred in August and averaged 46.2% across species and sites. Nitrogen and phosphorus mineralization rates also varied significantly from species to species. T. tomentosa having higher nitrogen content and lower C/N ratio than other species exhibited faster weight loss. Nitrogen and phosphorus contents of litter showed significant positive correlation with weight loss. C/N ratio was negatively related to decay constant, and the weight loss was positively related to the soil surface temperature as well as soil moisture content.     

Chemoprevention of Skin Cancer with 1,1-Bis (3′-Indolyl)-1-(Aromatic) Methane Analog through Induction of the Orphan Nuclear Receptor,NR4A2 (Nurr1)

Background

The objective of this study was to demonstrate the anti-skin cancer and chemopreventive potential of 1,1-bis(3′-indolyl)-1-(p-chlorophenyl methane) (DIM-D) using an in vitro model.

Methods

In vitro cell cytotoxicity and viability assays were carried out in A431 human epidermoid carcinoma cell line and normal human epidermal keratinocytes (NHEK) respectively by crystal violet staining. Apoptosis induction in A431 cells (DIM-D treated) and NHEK cells pretreated with DIM-D (2 hr) prior to UVB irradiation, were assessed. The accumulation of reactive oxygen species (ROS) in DIM-D pretreated NHEK cells (2 hr) prior to UVB exposure was also determined. Immunocytochemistry and western blot analysis was performed to determine cleaved caspase 3 and DNA damage markers in DIM-D treated A431 cells and in DIM-D pretreated NHEK cells prior to UVB irradiation.

Results

The IC50 values of DIM-D were 68.7±7.3, 48.3±10.1 and 11.5±3.1 μM whilst for Epigallocatechin gallate (EGCG) were 419.1±8.3, 186.1±5.2 and 56.7±3.1 μM for 24, 48 and 72 hr treatments respectively. DIM-D exhibited a significantly (p<0.05) greater induction of DNA fragmentation in A431 cells compared to EGCG with percent cell death of 38.9. In addition, DIM-D induced higher expression in A431 cells compared to EGCG of cleaved caspase 3 (3.0-fold vs. 2.4-fold changes), Nurr1 (2.7-fold vs. 1.7-fold changes) and NFκB (1.3-fold vs. 1.1-fold changes). DIM-D also exhibited chemopreventive activity in UVB-irradiated NHEK cells by significantly (p<0.05) reducing UVB-induced ROS formation and apoptosis compared to EGCG. Additionally, DIM-D induced expression of Nurr1 but reduced expression of 8-OHdG significantly in UVB-irradiated NHEK cells compared to EGCG and UV only.

Conclusion

Our results suggest that DIM-D exhibits Nurr1-dependent transactivation in the induction of apoptosis in A431 cells and it protects NHEK cells against UVB-induced ROS formation and DNA damage.     

High-Resolution Mutational Profiling Suggests the Genetic Validity of Glioblastoma Patient-Derived Pre-Clinical Models

Recent advances in the ability to efficiently characterize tumor genomes is enabling targeted drug development, which requires rigorous biomarker-based patient selection to increase effectiveness. Consequently, representative DNA biomarkers become equally important in pre-clinical studies. However, it is still unclear how well these markers are maintained between the primary tumor and the patient-derived tumor models. Here, we report the comprehensive identification of somatic coding mutations and copy number aberrations in four glioblastoma (GBM) primary tumors and their matched pre-clinical models: serum-free neurospheres, adherent cell cultures, and mouse xenografts. We developed innovative methods to improve the data quality and allow a strict comparison of matched tumor samples. Our analysis identifies known GBM mutations altering PTEN and TP53 genes, and new actionable mutations such as the loss of PIK3R1, and reveals clear patient-to-patient differences. In contrast, for each patient, we do not observe any significant remodeling of the mutational profile between primary to model tumors and the few discrepancies can be attributed to stochastic errors or differences in sample purity. Similarly, we observe ∼96% primary-to-model concordance in copy number calls in the high-cellularity samples. In contrast to previous reports based on gene expression profiles, we do not observe significant differences at the DNA level between in vitro compared to in vivo models. This study suggests, at a remarkable resolution, the genome-wide conservation of a patient’s tumor genetics in various pre-clinical models, and therefore supports their use for the development and testing of personalized targeted therapies.     

Human Granuloma In Vitro Model,for TB Dormancy and Resuscitation

Tuberculosis (TB) is responsible for death of nearly two million people in the world annually. Upon infection, Mycobacterium tuberculosis (Mtb) causes formation of granuloma where the pathogen goes into dormant state and can live for decades before resuscitation to develop active disease when the immune system of the host is weakened and/or suppressed. In an attempt to better understand host-pathogen interactions, several groups have been developing in vitro models of human tuberculosis granuloma. However, to date, an in vitro granuloma model in which Mtb goes into dormancy and can subsequently resuscitate under conditions that mimic weakening of the immune system has not been reported. We describe the development of a biomimetic in vitro model of human tuberculosis granuloma using human primary leukocytes, in which the Mtb exhibited characteristics of dormant mycobacteria as demonstrated by (1) loss of acid-fastness, (2) accumulation of lipid bodies (3) development of rifampicin-tolerance and (4) gene expression changes. Further, when these micro granulomas were treated with immunosuppressant anti-tumor necrosis factor-alpha monoclonal antibodies (anti-TNFα mAbs), resuscitation of Mtb was observed as has been found in humans. In this human in vitro granuloma model triacylglycerol synthase 1deletion mutant (Δtgs1) with impaired ability to accumulate triacylglycerides (TG), but not the complemented mutant, could not go into dormancy. Deletion mutant of lipY, with compromised ability to mobilize the stored TG, but not the complemented mutant, was unable to come out of dormancy upon treatment with anti-TNFα mAbs. In conclusion, we have developed an in vitro human tuberculosis granuloma model that largely exhibits functional features of dormancy and resuscitation observed in human tuberculosis.     

The Effect of N-acetylation and N-methylation of Lysine Residue of Tat Peptide on its Interaction with HIV-1 TAR RNA

Post-translational modification (PTM) of RNA binding proteins (RBPs) play a very important role in determining their binding to cognate RNAs and therefore regulate the downstream effects. Lysine can undergo various PTMs and thereby contribute to the regulation of different cellular processes. It can be reversibly acetylated and methylated using a pool of respective enzymes, to act as a switch for controlling the binding efficiency of RBPs. Here we have delineated the thermodynamic and kinetic effects of N-acetylation and N-monomethylation of lysine on interaction between HIV-1 TAR RNA and its cognate binder Tat peptide ( a model system). Our results indicate that acetylation of lysine 50 (K50), leads to eight- fold reduction in binding affinity, originating exclusively from entropy changes whereas, lysine 51 (K51) acetylation resulted only in three fold decrease with large enthalpy-entropy compensation. The measurement of kinetic parameters indicated major change (4.5 fold) in dissociation rate in case of K50 acetylation however, K51 acetylation showed similar effect on both association and dissociation rates. In contrast, lysine methylation did not affect the binding affinity of Tat peptide to TAR RNA at K50, nonetheless three fold enhancement in binding affinity was observed at K51 position. In spite of large enthalpy-entropy compensation, lysine methylation seems to have more pronounced position specific effect on the kinetic parameters. In case of K50 methylation, simultaneous increase was observed in the rate of association and dissociation leaving binding affinity unaffected. The increased binding affinity for methylated Tat at K51 stems from faster association rate with slightly slower dissociation rate.     

Impaired male meiosis, morphology and distribution pattern of different cytotypes of Bupleurum lanceolatum Wall. (Apiaceae) from the Western Himalayas

Detailed male meiosis, critical morphological observations and distribution pattern of diploid as well as tetraploid cytotypes of the Western Himalayan species, Bupleurum lanceolatum have been evaluated at present. A diploid (n = 8) cytotype is reported from Kashmir, whereas, both diploid (n = 8) and tetraploid (n = 16) cytotypes are available from two districts Kangra and Sirmaur of Himachal Pradesh. Out of these, the tetraploid cytotype makes new addition for the species on a worldwide basis. As per behavior, a tetraploid cytotype is characterized by abnormal meiosis leading to high pollen sterility and size variation of the pollen grains. Morphologically, tetraploids are noted to be luxuriant in comparison to the diploids.     

Cytological studies of Brassicaceae Burn. (Cruciferae Juss.) from Western Himalayas

Cytological studies have been carried out on 12 species of Brassicaceae Burn. on population basis from different geographical areas of Kashmir and Himachal Pradesh in the Western Himalayas. Variable chromosome reports for Barbaraea intermedia (n = 16), Cardamine loxostemonoides (n = 8), Nasturtium officinale (n = 8), Sisymbrium orientale (n = 14) on world-wide basis have been added to the previous reports of these species. The chromosome numbers in seven species as Barbaraea intermedia (n = 8), B. vulgaris (n = 8), Capsella bursa-pastoris (n = 8), Descuriania Sophia (n = 10), Rorippa islandica (n = 8), Sisymbrium strictum (n = 7) and Thlaspi alpestre (n = 7) have been worked out for the first time from India. The meiotic course in the populations of seven species such as Barbaraea intermedia, Capsella bursa-pastoris, Coronopus didymus, Descuriania sophia, Nasturtium officinale, Sisymbrium orientale and S. strictum varies from normal to abnormal while all the populations of two species Barbaraea vulgaris and Sisymbrium irio show abnormal meiotic course. Meiotic abnormalities are in the form of cytomixis, chromosomal stickiness, unoriented bivalents, inter-bivalent connections, formation of laggards and bridges, all resulting into abnormal microsporogenesis. Heterogenous sized fertile pollen grains and reduced reproductive potentialities have invariably been observed in all the meiotically abnormal populations. However, the meiotic course in all the populations of Cardamine loxostemonoides, Rorippa islandica and Thalspi alpestre is found to be normal with high pollen fertility.