Semiochemical based integrated livestock pest control.

The role of arthropods as livestock pests has been well established. Besides their biting habits causing nuisance in animals; they are important vectors for transmission of economically important livestock diseases worldwide. Various pests and vector control managemental programs that also make use of chemicals have variable success rates. Consequently, insecticide/acaricide resistance has been reported against most of the commonly used chemicals along with increased concern for environment and demand for clean and green, residue-free animal products. This calls for an urgent need to develop novel, alternate, effective strategies/technologies. This lays the foundation for the use of semiochemicals as alternatives along with other biological control agents. Current knowledge on semiochemical use in livestock is refined and limited; however, it has been widely exploited in the agricultural sector to control plant and food crop pests, surveillance, and monitoring. Semiochemicals have an added advantage of being natural and safe; however, knowledge of extraction and quantification by using assays needs to be explicit. Expertise is required in behavioral and electrophysiological studies of arthropods and their interactions with the host and environment targeting specific semiochemicals for promising results. A thorough prior understanding on aspects such as mechanism of action, the stimulus for the release, the effecter/target species, response produced, application methods, dose and concentration is required to develop any successful pest/vector control program. The current review provides essential and frontline information on semiochemicals and their potential applications in the livestock sector along with future challenges and interventions.

Bis-triazole linked organosilane based sensing platform for Cu2+ ions and insilico tyrosinase inhibitor activity.

The development of a ligand for their selective and sensitive detection is required due to the widespread use of Cu2+ in many industrial processes and the potential threat to human health. Herein, we report a bis-triazole linked organosilane (5) derived from the Cu(I) catalyzed azide-alkyne cycloaddition reaction. The synthesized compound 5 was characterized by (1H and 13C) NMR spectroscopic and mass spectrometry techniques. The UV-Visible and Fluorescence experiments of the designed compound 5 were performed with various metal ions, revealing its high selectivity and sensitivity to Cu2+ ions in MeOH: H2O (8:2, v/v, pH = 7.0, PBS buffer) solution. The selective fluorescence quenching upon addition of Cu2+ to the compound 5 is due to Photo-induced electron transfer process (PET). The limit of detection of compound 5 to Cu2+ was calculated as 2.56 × 10-6 M and 4.36 × 10-7 M through UV-Visible and Fluorescence titration data, respectively. The possible mechanism of 1:1 binding of 5 with Cu2+ could be affirmed by the density functional theory (DFT). Further, it was found that compound 5 showed a reversible behavior towards Cu2+ ions by the accumulation of sodium salt of CH3COO- which can be used in the construction of molecular logic gate where Cu2+ and CH3COO- are considered as inputs and the absorbance at 260 nm as output. Moreover, the molecular docking studies provide useful information about compound 5's interaction with the tyrosinase enzyme (PDB ID- 2Y9X).

Correlation of age at seizure onset with GABAA receptor subunit and chloride Co-transporter configuration in Focal cortical dysplasia (FCD).

Focal cortical dysplasia (FCD) represents a group of malformations of cortical development, which are speculated to be related to early developmental defects in the cerebral cortex. According to dysmature cerebral development hypothesis of FCD altered GABAA receptor function is known to contribute to abnormal neuronal network. Here, we studied the possible association between age at seizure onset in FCD with the subunit configuration of GABAA receptors in resected brain specimens obtained from patients with FCD. We observed a significantly higher ratio of α4/α1 subunit-containing GABAA receptors in patients with early onset (EO) FCD as compared to those with late onset (LO) FCD as is seen during the course of development where α4-containing GABAA receptors expression is high as compared to α1-containing GABAA receptors expression. Likewise, the influx to efflux chloride co-transporter expression of NKCC1/KCC2 was also increased in patients with EO FCD as seen during brain development. In addition, we observed that the ratio of GABA/Glutamate neurotransmitters was lower in patients with EO FCD as compared to that in patients with LO FCD. Our findings suggest altered configuration of GABAA receptors in FCD which could be contributing to aberrant depolarizing GABAergic activity. In particular, we observed a correlation of age at seizure onset in FCD with subunit configuration of GABAA receptors, levels of NKCC1/KCC2 and the ratio of GABA/Glutamate neurotransmitters such that the patients with EO FCD exhibited a more critically modulated GABAergic network.

Identification and Manipulation of Defects in Black Phosphorus.

We identify and manipulate commonly occurring defects in black phosphorus, combining scanning tunneling microscopy experiments with density functional theory calculations. A ubiquitous defect, imaged at negative bias as a bright dumbbell extending over several nanometers, is shown to arise from a substitutional Sn impurity in the second sublayer. Another frequently observed defect type is identified as arising from an interstitial Sn atom; this defect can be switched to a more stable configuration consisting of a Sn substitutional defect + P adatom, by application of an electrical pulse via the STM tip. DFT calculations show that this pulse-induced structural transition switches the system from a non-magnetic configuration to a magnetic one. We introduce States Projected Onto Individual Layers (SPOIL) quantities which provide information about atom-wise and orbital-wise contributions to bias-dependent features observed in STM images.

Blue and black phosphorene on metal substrates: a density functional theory study.

We have performed density functional theory calculations to study blue phosphorene and black phosphorene on metal substrates. The substrates considered are the (111) and (110) surfaces of Al, Cu, Ag, Ir, Pd, Pt and Au and the (0001) and (101¯0) surfaces of Zr and Sc. The formation energyEFis negative (energetically favorable) for all 36 combinations of overlayer and substrate. By comparing values of ΔΩ, the change in free energy per unit area, as well as the overlayer-substrate binding energyEb, we identify that Ag(111), Al(110), Cu(111), Cu(110) and possibly Au(110) may be especially suitable substrates for the synthesis and subsequent exfoliation of blue phosphorene, and the Ag(110) and Al(111) substrates for the synthesis of black phosphorene. However, these conclusions are drawn assuming the source of P atoms is bulk phosphorus, and can alter upon changing synthesis conditions (chemical potential of phosphorus). Thus, when the source of phosphorus atoms is P4, blue phosphorene is favored only over Pt(111). We find that for all combinations of overlayer and substrate, the charge transfer per bond can be captured by the universal descriptorD=Δχ/ΔR, where ΔχandΔRare, respectively, the differences in electronegativity and atomic size between phosphorus and the substrate metal.

Transcriptomic profiling of high- and low-spiking regions reveals novel epileptogenic mechanisms in focal cortical dysplasia type II patients.

Focal cortical dysplasia (FCD) is a malformation of the cerebral cortex with poorly-defined epileptogenic zones (EZs), and poor surgical outcome in FCD is associated with inaccurate localization of the EZ. Hence, identifying novel epileptogenic markers to aid in the localization of EZ in patients with FCD is very much needed. High-throughput gene expression studies of FCD samples have the potential to uncover molecular changes underlying the epileptogenic process and identify novel markers for delineating the EZ. For this purpose, we, for the first time performed RNA sequencing of surgically resected paired tissue samples obtained from electrocorticographically graded high (MAX) and low spiking (MIN) regions of FCD type II patients and autopsy controls. We identified significant changes in the MAX samples of the FCD type II patients when compared to non-epileptic controls, but not in the case of MIN samples. We found significant enrichment for myelination, oligodendrocyte development and differentiation, neuronal and axon ensheathment, phospholipid metabolism, cell adhesion and cytoskeleton, semaphorins, and ion channels in the MAX region. Through the integration of both MAX vs non-epileptic control and MAX vs MIN RNA sequencing (RNA Seq) data, PLP1, PLLP, UGT8, KLK6, SOX10, MOG, MAG, MOBP, ANLN, ERMN, SPP1, CLDN11, TNC, GPR37, SLC12A2, ABCA2, ABCA8, ASPA, P2RX7, CERS2, MAP4K4, TF, CTGF, Semaphorins, Opalin, FGFs, CALB2, and TNC were identified as potential key regulators of multiple pathways related to FCD type II pathology. We have identified novel epileptogenic marker elements that may contribute to epileptogenicity in patients with FCD and could be possible markers for the localization of EZ.

Differential regulation of excitatory synaptic transmission in the hippocampus and anterior temporal lobe by cyclin dependent kinase 5 (Cdk5) in mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS).

Mesial temporal lobe epilepsy with hippocamapal sclerosis (MTLE-HS) is the most common form of drug resistant epilepsy (DRE). MTLE-HS is a distributed network disorder comprising of not only the hippocampus, but other anatomically related extrahippocampal regions. Excitatory synaptic transmission is differentially regulated in the hippocampal and extra-hippocampal regions of patients with MTLE-HS, but its mechanism not understood. Cyclin-dependent kinase 5 (Cdk5) is known to regulate synaptic transmission and plasticity through up-regulation of NMDA receptors by phosphorylating NR2Asubunits. The present study is designed to investigate whether Cdk5 differentially regulates the excitatory synaptic transmission in the hippocampus and anterior temporal lobe (ATL) samples obtained from patients of MTLE-HS. We have measured the Cdk5 kinase activity and the protein levels of Cdk5, p-Cdk5, p35/p25, NR2A, pNR2A in the hippocampal and ATL samples obtained from patients with MTLE-HS. We have also determined the effect of roscovitine, a Cdk5 antagonist, on spontaneous excitatory postsynaptic currents (EPSCs) recorded from the hippocampal and ATL using patch-clamp technique. We observed significant increase in the expression of Cdk5, p-Cdk5, p35/p25, NR2A, pNR2A in the ATL samples as compared to the hippocampal samples. Cdk5 activity was significantly higher in ATL samples as compared to the hippocampal samples. Magnitude of reduction in the frequency of EPSCs by roscovitine in the ATL samples was higher than that in the hippocampal samples. Our studies suggest that Cdk5 differentially regulates excitatory synaptic activity in the hippocampal and ATL region of patients with MTLE-HS.

Increased levels of α4-containing GABAA receptors in focal cortical dysplasia: A possible cause of benzodiazepine resistance.

Benzodiazepines are the first choice of anti-epileptic drugs used to treat seizures. However, it has been seen that their efficacy decreases with time leading to drug insensitivity, plausibly caused by an alteration in the expression of the benzodiazepine biding site on GABAA receptors. This study was designed to investigate if the differential expression of GABAA receptor subunits α1/α4/γ2/δ across the postsynaptic sites could contribute to benzodiazepine resistance in patients with focal cortical dysplasia (FCD), the most common cause of drug resistant epilepsy in pediatric population. Differential gene and cellular expression of GABAA receptor subunits α1, α4, γ2 and δ were evaluated and validated using qPCR and immunohistochemistry. Whole cell patch clamp studies were performed on pyramidal neurons of resected cortical FCD samples to measure the spontaneous GABAA receptor activity. Upregulation of α4-and γ2-subunits containing GABAA receptors were observed at both mRNA and protein level. α1-and δ-subunits containing GABAA receptors did not show any significant changes. Flumazenil treatment did not affect the kinetics of GABAergic events in FCD; however, it significantly reduced the frequency and amplitude of spontaneous GABAergic activity in non-seizure control samples. Our results demonstrate the enhanced expression of α4-containing GABAA receptors and GABAergic activity in pyramidal neurons which in turn may contribute to benzodiazepine resistance in FCD patients.

Integrated Genome-Wide DNA Methylation and RNAseq Analysis of Hippocampal Specimens Identifies Potential Candidate Genes and Aberrant Signalling Pathways in Patients with Hippocampal Sclerosis.

BACKGROUND AND AIMS: DNA methylation and demethylation play a crucial role in the regulation of gene expression, though their interplay during pathogenesis of hippocampal scelerosis (HS) remains elusive. The present study was designed to investigate the DNA methylation regulated changes in expression of HS patients. METHODS: We performed integrative analysis of genome-wide CpG-DNA methylation profiling and RNA sequencing to profile global changes in promoter methylation and gene expression in HS patients. Real time PCR was performed to validate the findings of methylation and RNA sequencing. RESULTS: A total of 16040 sites showed altered DNA methylation in all the CpG islands. Of these, 3185 sites were in the promoter regions, of which 66 genes showed an inverse correlation between methylation and expression. These genes are largely related to pathways predicted to participate in axon guidance by semaphorins, MAPK, ionotropic glutamate receptor pathway, notch signaling, regulatory activities related to TFAP2A and immune response, with the most distinct ones included TFAP2A, NRP1, SEMA3B, CACNG2, MAP3K11, and ADAM17. CONCLUSION: We performed integrated analysis of genomic methylation signature and differential gene expression patterns of hippocampal tissues resected from patients with HS for the first time. Collectively, our findings implicate DNA methylation as a critical regulator of the pathogenic mechanisms of epileptogenesis associated with HS.

Genome-wide DNA Methylation and RNAseq Analyses Identify Aberrant Signalling Pathways in Focal Cortical Dysplasia (FCD) Type II.

Focal cortical dysplasia (FCD) is one of the most common pathologies associated with drug-resistant epilepsy (DRE). The pharmacological targets remain obscured, as the molecular mechanisms underlying FCD are unclear. Implications of epigenetically modulated aberrant gene expression in disease progression are reported in various DRE pathologies except FCD. Here we performed genome-wide CpG-DNA methylation profiling by methylated DNA immunoprecipitation (MeDIP) microarray and RNA sequencing (RNAseq) on cortical tissues resected from FCD type II patients. A total of 19088 sites showed altered DNA methylation in all the CpG islands. Of these, 5725 sites were present in the promoter regions, of which 176 genes showed an inverse correlation between methylation and gene expression. Many of these 176 genes were found to belong to a cohesive network of physically interacting proteins linked to several cellular functions. Pathway analysis revealed significant enrichment of receptor tyrosine kinases (RTK), EGFR, PDGFRA, NTRK3, and mTOR signalling pathways. This is the first study that investigates the epigenetic signature associated with FCD type II pathology. The candidate genes and pathways identified in this study may play a crucial role in the regulation of the pathogenic mechanisms of epileptogenesis associated with FCD type II pathologies.

Discrimination, molecular characterisation and phylogenetic comparison of porcine Eimeria spp. in India.

Infections with Eimeria spp. are common in pigs worldwide, occasionally affecting animals clinically after weaning or during the fattening period when diarrhoea and weight loss can be observed upon infection with the more pathogenic species. Molecular characterization of pathogens is valuable to accurately delimit species and development novel diagnostics, although sequences which define Eimeria species that infect pigs are scarce. Only three of the eight common species are currently represented in GenBank. In this study we describe the occurrence of Eimeria species in pigs sampled in Punjab, India; going on to use the samples to generate new species-specific 18S rDNA sequences for all of the previously uncharacterised species. Using these data we report the first phylogenetic analyses to include the eight Eimeria species that commonly infect the domestic pig (Sus scrofa domesticus). Consideration of phylogenetic trees produced using Maximum Likelihood, Neighbour Joining, Maximum Parsimony and Unweighted Pair Group Method with Arithmetic Mean methods indicate that the 18S rDNA sequences present lower levels of genetic diversity than Eimeria which infect avian species and are insufficient to infer stable phylogenies.

Upregulation of breast cancer resistance protein and major vault protein in drug resistant epilepsy.

PURPOSE: Identifying factors involved in the development of drug resistant epilepsy (DRE) remains a challenge. Candidate gene studies have shown modulation of resistance to drugs by various multidrug resistance proteins in DRE. However the resistance to drugs in DRE could be more complex and multifactorial involving molecules in different pharmacokinetic processes. In this study for the first time we have analyzed the relative expression of four molecules with different drug resistance mechanisms in two most common DRE pathologies, mesial temporal lobe epilepsy (MTLE) and focal cortical dysplasia (FCD) with respect to each other and also with different non-epileptic controls. METHODS: Brain tissues resected from MTLE (n=16) and FCD type I and II (n=12) patients who had undergone surgery were analysed for mRNA levels of multidrug resistance-associated protein 1(MRP1), major vault protein (MVP), breast cancer resistance protein (BCRP), and one drug metabolising enzyme (UGT1A4) as compared to non-epileptic controls which were tissues resected from tumor periphery (n=6) and autopsy tissues (n=4) by quantitative PCR. RESULTS: We found significant upregulation of MVP and BCRP whereas MRP1 and UGT1A4 were unaltered in both pathologies. While upregulation of BCRP was significantly higher in MTLE (9.34±0.45; p<0.05), upregulation of MVP was significantly higher in FCD (2.94±0.65; p<0.01). CONCLUSION: We propose that upregulation of BCRP and MVP is associated with MTLE and FCD and these molecules not only may have the potential to predict pathology specific phenotypes but may also have therapeutic potential as adjunct treatment in these pathologies.

Prevalence of gastrointestinal parasites in dogs of Palampur, Himachal Pradesh.

A total of 246 faecal/scat samples of the dogs were screened by direct and floatation concentration technique to study the gastrointestinal (GI) tract parasitism in dogs of Palampur, Himachal Pradesh, India. Detailed coprological examination targeting different seasons, age groups and living styles of the dogs revealed an overall 28.04 % of GI parasitism with highest prevalence in summer season (37.87 %). Stray dogs harbored 47.29 % GI parasites in comparison to 19.19 % of pet dogs. Highest prevalence of GI parasitism was observed in the pups, below 3 months of age (39.13 %), followed by the dogs with the age ranging from 3 months to 1 year (26.38 %) and lowest in dogs of the age ranging from 1 to 3 years (6.77 %). Amongst all the parasites, Toxocara canis (44.93 %) infection was highest, followed by Dipylidium caninum (17.39 %) and hookworms (15.94 %).