Homology among the autosomal chromosomes of \u3ci\u3eBoophilus annulatus\u3c/i\u3e (Say) and \u3ci\u3eB. microplus\u3c/i\u3e (Canestrini)

Autosomes of Boophilus annulatus and B. microplus were compared using C- and G-bands to establish the degree of homology. Karyotypes of both species consisted of 20 autosomes and an XX:XO sex determination system with the X being the largest chromosome. All chromosomes of B. annulatus were acrocentric with heterochromatin limited to centromeric regions. The B. microplus karyotype was acrocentric with one band of noncentromeric heterochromatin occurring in three chromosome pairs. Interspecific comparisons indicated seven pairs of G-band homologous autosomes and three G-band homologous pairs when added interstitial heterochromatin was considered. The number three chromosome of both species showed variation in length at an area of extreme constriction associated with the nucleolar organizer region

A Study of Deoxyribooligonucleotide Duplexes Containing an Unpaired Base or Mismatched Base Pairs Using: (I) Molecular Modeling, And, (II) Proton Exchange Measured With Nuclear Magnetic Resonance Spectroscopy.

Mismatched base pairs have been implicated in mutations and arise from transitions or transversions of the base pairs during replication. An oligonucleotide duplex containing adjacent G$\cdot$A and A$\cdot$A mismatches, d(GCGAAT)$\cdot$d(ATAAGC), has been investigated using molecular mechanics and dynamics. Experimental NMR data obtained for a related molecule was used to constrain the model which suggested an unusual hydrogen-bonding for both mismatches. Between the G$\cdot$A and A$\cdot$A mismatches, defined by the unusual base pairing, there is a large twist angle. The remainder of the base pairs are B-DNA conformations. The effect of an unpaired base on the local geometry of an oligonucleotide duplex containing an (oligo dA)$\cdot$(oligo dT) tract was investigated using molecular modeling. The following duplexes have been studied: \vbox{\halign{#\hfil&&\qquad#\hfil\cr &dGCGAA{\bf X}AAGCG &dGCGAA-AAGCG &dGCGAAAACGC\cr &\ CGCTT-TTCGCd &CGCTT{\bf Y}TTCGCd &CGCTTTTCGCd\cr}}where X is cytosine, thymine or guanine, and where Y is guanine, adenine or thymine. The structures are of interest because they contain an A-tract, which is implicated in DNA bending and a region that is a model for mutational hot spots. The models were built with the unpaired base in two possible orientations, extrahelical and intrahelical. The results indicate that in the intrahelical conformation the flanking base pairs form a wedge and in the extrahelical conformation the flanking base pairs remain in a B-DNA stack. The extrahelical pyrimidines are in the minor groove whereas the extrahelical purines have two possible conformations either in the major groove or in the minor groove. The effect of an unpaired base on the base-pair opening rate of an oligonucleotide duplex containing an (oligo dA)$\cdot$(oligo dT) tract has been investigated using proton exchange NMR techniques on: \vbox{\halign{#\hfil&&\qquad#\hfil\cr &dGCGAA{\bf \ G}AAGCG &dGCGAA-AAGCG &dGCGAAAACGC\cr &\ CGCTT-TTCGCd & CGCTT{\bf \ G}TTCGCd &CGCTTTTCGCd\cr}}For d(GCGAAGAAGCG)$\cdot$d(CGCTTTTCGC) the base pairs adjacent to the unpaired G have faster opening rates than do those base pairs further away. For d(GCGAAAAGCG)$\cdot$d(CGCTTGTTCGC) the base pairs adjacent to the unpaired base have very fast opening rates. The activation energies are similar for d(GCGAAAAGCG)$\cdot$d(CGCTTTTCGC), d(GCGAAGAAGCG)$\cdot$d(CGCTTTTCGC) and d(GCGAAAAGCG)$\cdot$d(CGCTTGTTCGC)

The FOXO1 inhibitor AS1842856 triggers apoptosis in glioblastoma multiforme and basal-like breast cancer cells

Basal-like breast cancer (BBC) and glioblastoma multiforme (GBM) are poor-prognosis cancers that lack effective targeted therapies and harbor embryonic stem gene expression signatures. Recently, our group and others found that forkhead box transcription factor FOXO1 promotes stem gene expression in BBC and GBM cell lines. Given the critical role of cancer stem cells in promoting cancer progression, we examined the impact of FOXO1 inhibition with AS1842856 (a cell-permeable small molecule that directly binds to unphosphorylated FOXO1 protein to block transcriptional regulation) on BBC and GBM cell viability. We treated a set of BBC and GBM cancer cell lines with increasing concentrations of AS1842856 and found reduced colony formation. Treatment of BBC and GBM cancer cells with AS1842856 led to increases in FAS (FAS cell surface death receptor) and BIM (BCL2L11) gene expression, as well as increased positivity for markers for apoptosis such as annexin V and propidium iodide. Treatment with another FOXO1 inhibitor AS1708727 or FOXO1 RNAi also led to FAS induction. This work is the first to show that targeting BBC and GBM with FOXO1 inhibition leads to apoptosis. These novel findings may ultimately expand the repertoire of therapies for poor-prognosis cancers

Effects of Concurrent Selection for Residual Feed Intake and Average Daily Gain on Fertility and Longevity in Black Angus Beef Females

Data from this analysis suggest that concurrent selection for both average daily gain and residual feed intake (RFI) may identify beef heifers that have improved fertility and longevity without impacting growth and maternal EPDs. As the beef industry continues to focus on sustainability, and thus efficiency, identification of commercial breeding stock that fit this mold will be imperative

PI3K Pathway Inhibition with NVP-BEZ235 Hinders Glycolytic Metabolism in Glioblastoma Multiforme Cells

Glioblastoma (GBM) is the most lethal primary brain cancer that lacks effective molecular targeted therapies. The PI3K/AKT/mTOR pathway is activated in 90% of all Glioblastoma multiforme (GBM) tumors. To gain insight into the impact of the PI3K pathway on GBM metabolism, we treated U87MG GBM cells with NVP-BEZ235 (PI3K and mTOR a dual inhibitor) and identified differentially expressed genes with RNA-seq analysis. RNA-seq identified 7803 differentially regulated genes in response to NVP-BEZ235. Gene Set Enrichment Analysis (GSEA) identified two glycolysis-related gene sets that were significantly enriched (p \u3c 0.05) in control samples compared to NVP-BEZ235-treated samples. We validated the inhibition of glycolytic genes by NVP-BEZ235 and examined the impact of the FOXO1 inhibitor (AS1842856) on these genes in a set of GBM cell lines. FOXO1 inhibition alone was associated with reduced LDHA expression, but not ENO1 or PKM2. Bioinformatics analyses revealed that PI3K-impacted glycolytic genes were over-expressed and co-expressed in GBM clinical samples. The elevated expression of PI3K-impacted glycolytic genes was associated with poor prognosis in GBM based on Kaplan–Meier survival analyses. Our results suggest novel insights into hallmark metabolic reprogramming associated with the PI3K-mTOR dual inhibition

FOXO1 promotes the expression of canonical WNT target genes in examined basal-like breast and glioblastoma multiforme cancer cells

Basal-like breast cancer (BBC) and glioblastoma multiforme (GBM) are aggressive cancers associated with poor prognosis. BBC and GBM have stem-cell-like gene expression signatures, which are in part driven by forkhead box O (FOXO) transcription factors. To gain further insight into the impact of FOXO1 in BBC, we treated BT549 cells with AS1842856 and performed RNA sequencing. AS1842856 binds to unphosphorylated FOXO1 and inhibits its ability to directly bind to DNA. Gene Set Enrichment Analysis (GSEA) indicated that a set of WNT pathway target genes, including lymphoid enhancer-binding factor 1 (LEF1) and transcription factor 7 (TCF7), were robustly induced after AS1842856 treatment. These same genes were also induced in GBM cell lines U87MG, LN18, LN229, A172 and DBTRG upon AS1842856 treatment. In contrast, follow-up RNA interference (RNAi) targeting of FOXO1 led to reduced LEF1 and TCF7 gene expression in BT549 and U87MG cells. In agreement with RNAi experiments, CRISPR Cas9-mediated FOXO1 disruption reduced the expression of canonical WNT genes LEF1 and TCF7 in U87MG cells. The loss of TCF7 gene expression in FOXO1 disruption mutants was restored by exogenous expression of the DNA-binding-deficient FOXO1-H215R. Therefore, FOXO1 induces TCF7 in a DNA-binding-independent manner, similar to other published FOXO1-activated genes such as TCF4 and hes family bHLH transcription factor 1 (HES1). Our work demonstrates that FOXO1 promotes canonical WNT gene expression in examined BBC and GBM cells, similar to results found in Drosophila melanogaster, T-cell development and murine acute myeloid leukemia (AML) models

Epidemic infectious gastrointestinal illness aboard U.S. Navy ships deployed to the Middle East during peacetime operations – 2000–2001

BACKGROUND: Infectious gastrointestinal illness (IGI) outbreaks have been reported in U.S. Navy ships and could potentially have an adverse mission impact. Studies to date have been anecdotal. METHODS: We conducted a retrospective analysis of weekly reported disease and non-battle injury health data collected in 2000 – 2001 from 44 U.S. Navy ships while sailing in the 5(th )Fleet (Persian Gulf and nearby seas). RESULTS: During this period, 11 possible IGI outbreaks were identified. Overall, we found 3.3 outbreaks per 100 ship-weeks, a mean outbreak duration of 4.4 weeks, and a mean cumulative ship population attack rate of 3.6%. Morbidity, represented by days lost due to personnel being placed on sick-in-quarters status, was higher during outbreak weeks compared to non-outbreak weeks (p = 0.002). No clear seasonal distribution was identified. CONCLUSION: Explosive outbreaks due to viruses and bacteria with the potential of incapacitating large proportions of the crew raise serious concerns of mission impact and military readiness

The Determination of Biological Activity and Biochemical Mode of Action for the Oxadiazole and Diacylhdrazine Insecticides

This report includes the determination of activity and possible mode of action for a group of potential new insecticides. The biological screening procedure was developed using Drosophila melanogaster as the test organism while Musca domestica was used for mode of action assays. The percent kill for each compound is based on the number of eggs placed on media containing the insecticide minus the number of adults enclosed as compared to the control media reared flies. Observations were made on all stages from eggs through adults to determine time of death and if any malformations were present. These observations aided in the mode of action studies as did preciously published work on diflurbenzuron. The mode of action studies encompassed chitin synthesis, chitin breakdown and DNA synthesis. Cuticle deposition was determined gravimetrically on pupal instar reared on media with and without DOWCO 416. Chitin synthesis and DNA synthesis were followed by measuring the incorporation of radiolabeled precursors by pupal instars reared on media with and without DOWCO 416. Chitin breakdown was followed through the measurement of chitinase activity spectrophotometrically on all stages of larvae which were reared on media with and without DOWCO 416. Direct inhibition of chitinase was investigated by incubation of the purified chitinase from Staphlycoccus griseus with varying concentrations of the test compound