Therapeutic resistance to gemcitabine in pancreatic ductal adenocarcinoma (PDAC) is attributed to various cellular mechanisms and signaling molecules that influence as a single factor or in ...combination.
In this study, utilizing in vitro p21-activated kinase 1 (Pak1) overexpression and knockdown cell line models along with in vivo athymic mouse tumor xenograft models and clinical samples, we demonstrate that Pak1 is a crucial signaling kinase in gemcitabine resistance.
Pak1 kindles resistance via modulation of epithelial–mesenchymal transition and activation of pancreatic stellate cells. Our results from gemcitabine-resistant and -sensitive cell line models showed that elevated Pak1 kinase activity is required to confer gemcitabine resistance. This was substantiated by elevated levels of phosphorylated Pak1 and ribonucleotide reductase M1 levels in the majority of human PDAC tumors when compared with normal. Delineation of the signaling pathway revealed that Pak1 confers resistance to gemcitabine by preventing DNA damage, inhibiting apoptosis and regulating survival signals via NF-κB. Furthermore, we found that Pak1 is an upstream interacting substrate of transforming growth factor β-activated kinase 1—a molecule implicated in gemcitabine resistance. Molecular mechanistic studies revealed that gemcitabine docks with the active site of Pak1; furthermore, gemcitabine treatment induces Pak1 kinase activity both in vivo and in cell-free system. Finally, results from athymic mouse tumor models illustrated that Pak1 inhibition by IPA-3 enhances the cytotoxicity of gemcitabine and brings about pancreatic tumor regression.
To our knowledge, this is the first study illustrating the mechanistic role of Pak1 in causing gemcitabine resistance via multiple signaling crosstalks, and hence Pak1-specific inhibitors will prove to be a better adjuvant with existing chemotherapy modality for PDAC.
Foot-and-mouth disease (FMD) is a contagious viral disease affecting cloven hoofed livestock. Insect cell expressed virus like particles (VLPs) are potential alternative to overcome the limitations ...of inactivated vaccine. However, at pH < 6.5, virus particles disassociate into pentameric structure resulting in loss of antigenicity. Accordingly, we generated seven mutant VLPs containing mutations in the structural genes of FMDV vaccine strains (N17D and/or H145Y for serotypes O/IND/R2/75 and Asia1/IND/63/72; and H142D for serotype A/IND/40/00) by PCR based site directed mutagenesis. Acid resistant VLPs produced by baculovirus expression system were tested for acid stability at pH 7.5, 6.5, 6.0 and 5.5 followed by reactivity in sandwich-ELISA (s-ELISA), which revealed mutant-1 (N17D) of serotype O and Asia1 retained the antigenicity in s-ELISA even at pH 5.5 as compared to other VLPs and wild-types. Further, the 75S empty capsids obtained in sucrose density gradient, when tested in liquid phase blocking ELISA (LPBE) in comparison to cell culture antigen indicated that the VLPs were stable at acidic pH. Transmission electron microscopy of OM-1 confirmed the intact morphology of the empty VLPs. It is concluded that acid resistant VLPs could be useful for developing new generation vaccine or diagnostic for FMDV.
Summary
The goal of this study was to characterize the properties and duration of the foot‐and‐mouth disease (FMD) carrier state and associated serological responses subsequent to vaccination and ...naturally occurring infection at two farms in northern India. Despite previous vaccination of cattle in these herds, clinical signs of FMD occurred in October 2013 within a subset of animals at the farms containing juvenile‐yearling heifers and steers (Farm A) and adult dairy cattle (Farm B). Subsequent to the outbreak, FMD virus (FMDV) asymptomatic carriers were identified in both herds by seroreactivity to FMDV non‐structural proteins and detection of FMDV genomic RNA in oropharyngeal fluid. Carriers’ seroreactivity and FMDV genome detection status were subsequently monitored monthly for 23 months. The mean extinction time of the carrier state was 13.1 ± 0.2 months, with extinction having occurred significantly faster amongst adult dairy cattle at Farm B compared to younger animals at Farm A. The rate of decrease in the proportion of carrier animals was calculated to be 0.07 per month. Seroprevalence against FMDV non‐structural proteins decreased over the course of the study period, but was found to increase transiently following repeated vaccinations. These data provide novel insights into viral and host factors associated with the FMDV carrier state under natural conditions. The findings reported herein may be relevant to field veterinarians and governmental regulatory entities engaged in FMD response and control measures.
Summary
Foot‐and‐mouth disease (FMD) is an important transboundary disease with substantial economic impacts. Although between‐herd transmission of the disease has been well studied, studies focusing ...on within‐herd transmission using farm‐level outbreak data are rare. The aim of this study was to estimate parameters associated with within‐herd transmission, host physiological factors and FMD virus (FMDV) persistence using data collected from an outbreak that occurred at a large, organized dairy farm in India. Of 1,836 regularly vaccinated, adult dairy cattle, 222 had clinical signs of FMD over a 39‐day period. Assuming homogenous mixing, a frequency‐dependent compartmental model of disease transmission was built. The transmission coefficient and basic reproductive number were estimated to be between 16.2–18.4 and 67–88, respectively. Non‐pregnant animals were more likely to manifest clinical signs of FMD as compared to pregnant cattle. Based on oropharyngeal fluid (probang) sampling and FMDV‐specific RT‐PCR, four of 36 longitudinally sampled animals (14%) were persistently infected carriers 10.5 months post‐outbreak. There was no statistical difference between subclinical and clinically infected animals in the duration of the carrier state. However, prevalence of NSP‐ELISA antibodies differed significantly between subclinical and clinically infected animals 12 months after the outbreak with 83% seroprevalence amongst clinically infected cattle compared to 69% of subclinical animals. This study further elucidates within‐herd FMD transmission dynamics during the acute‐phase and characterizes duration of FMDV persistence and seroprevalence of FMD under natural conditions in an endemic setting.
Summary
A systematic vaccination programme is ongoing in India to control the three prevailing serotypes (A, O, Asia1) of foot‐and‐mouth disease (FMD) virus. Under the programme, more than ...120 million bovine (term bovine applicable to both cattle and buffalo in this study) population of 221 of the 666 districts in the country are being bi‐annually vaccinated with trivalent vaccine since 2010. Although clinical disease has reduced in these districts because of the systematic vaccinations, an abrupt increase in the number of FMD cases was recorded in 2013. Hence, a longitudinal field study was conducted in the year 2014 to estimate the serological herd immunity level in bovines, the impact of systematic vaccinations and field efficacy of the vaccines used. Serum samples (n = 115 963) collected from 295 districts of the 18 states of the country were analysed to estimate antibody titres against structural proteins of the three serotypes. The efficacy of the vaccine was demonstrated in the control group (group‐D) where animals of the group were identified by ear tags for the purpose of repeated sampling after vaccination. Progressive building of the herd immunity in the field after systematic vaccination was demonstrated. The mean antibody titre against the serotypes O, A and Asia1 was estimated as log10 1.93 (95% CI 1.92–1.93), 2.02 (2.02–2.02) and 2.02 (2.02–2.02), respectively, in the states covered under the control programme. However, in other states herd immunity was significantly low mean titre log10 1.68 (95% CI 1.67–1.69), 1.77 (1.76–1.78) and 1.85 (1.84–1.86) against the three serotypes. Inverse relationship between the herd immunity and FMD incidences was observed the states following different vaccination practices. The study helped in demarcation of FMD risk zones in the country with low herd immunity. Estimation of herd immunity kinetics in the field helped in refining the vaccination schedule under the control programme.
Taxonomic ambiguity exists in genus Systomus and recently many new species were described under this genus. Systomus sarana subnasutus is considered a valid subspecies of S. sarana sarana although ...revisions have been done by some researchers. We employed a combination of morpho-meristics and molecular tools (Cytochrome c oxidase I, 16S and Cytochrome b genes of mitochondrial genome) to resolve the two species. Three morpho-meristic characters, head length/maxillary barbel length (HL/MxBL), Lateral Line Scales (LLSs) as well as two truss-based characters, had discernible variation between the two taxa. The sequence analysis (2353 nucleotides) depicted a separate clad of S. sarana subnasutus with high bootstrap support. The findings from combined use of morphology, meristics and mitogenes were concordant. The corroborative results suggest the possibility of two different species. The results suggest to adopt suitable management measures, accordingly.
Fused deposition modeling (FDM), an advanced 3D printing method, is a rapidly growing technology for realizing geometrically complex parts. FDM utilizes a thermoplastic feedstock that is heated and ...extruded through a nozzle to build up layers of material following toolpaths prescribed by three‐dimensional (3D) data. However, the rapid thermal cycles that the material undergoes during this process limit the formation, growth, and entanglement of the molecular chains in the material. Herein, we have proposed a novel laser‐assisted fused deposition modeling (LAFDM) process for the improvement in printed part properties such as bond width and tensile strength. The LAFDM system was developed consisting of an infrared CO2 laser, coupled with an industrial FDM printer. Using an adapted tensile testing method, the bond strength between deposited layers of acrylonitrile butadiene styrene (ABS) in 3D‐printed structures was assessed. Thermogravimetric analysis was used to monitor the thermal stability of the polymer to detect alterations to the chemical composition of the material as a result of the processing conditions. A 9.5% increase in the average bond tensile strength for the LAFDM‐printed specimen was observed compared to that of the non‐laser‐assisted FDM‐printed specimen. The improvement to the bond strengths was achieved without compromising the thermal stability of the polymer.
Systomus sarana sarana, commonly known as olive barb, is an important food and ornamental fish species with wide distribution in South Asia. Here, the complete mitogenome was sequenced on HiSeq 2500. ...With 16,590 nucleotides, the base composition was 32.9% (A), 26% (C), 15.4% (G) and 25.7% (T), comparable with other carps. The clustering pattern depicted the monophyly of S. sarana sarana with sister cyprinids.
With the full data sample of 772×106 BB¯ pairs recorded by the Belle detector at the KEKB electron-positron collider, the decay B¯→D*τ−ν¯τ is studied with the hadronic τ decays τ−→π−ντ and τ−→ρ−ντ. ...The τ polarization Pτ(D*) in two-body hadronic τ decays is measured, as well as the ratio of the branching fractions R(D*)=B(B¯→D*τ−ν¯τ)/B(B¯→D*ℓ−ν¯ℓ), where ℓ− denotes an electron or a muon. Our results, Pτ(D*)=−0.38±0.51(stat)−0.16+0.21(syst) and R(D*)=0.270±0.035(stat)−0.025+0.028(syst), are consistent with the theoretical predictions of the standard model. The polarization values of Pτ(D*)>+0.5 are excluded at the 90% confidence level.
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