A sumoylation program is essential for maintaining the mitotic fidelity in proliferating mantle cell lymphoma cells.

BACKGROUND: Mantle cell lymphoma (MCL) is a rare, highly heterogeneous type of B-cell non-Hodgkin's lymphoma. The sumoylation pathway is known to be upregulated in many cancers including lymphoid malignancies. However, little is known about its oncogenic role in MCL. METHODS: Levels of sumoylation enzymes and sumoylated proteins were quantified in MCL cell lines and primary MCL patient samples by scRNA sequencing and immunoblotting. The sumoylation enzyme SAE2 was genetically and pharmacologically targeted with shRNA and TAK-981 (subasumstat). The effects of SAE2 inhibition on MCL proliferation and cell cycle were evaluated using confocal microscopy, live-cell microscopy, and flow cytometry. Immunoprecipitation and orbitrap mass spectrometry were used to identify proteins targeted by sumoylation in MCL cells. RESULTS: MCL cells have significant upregulation of the sumoylation pathway at the level of the enzymes SAE1 and SAE2 which correlated with poor prognosis and induction of mitosis associated genes. Selective inhibition of SAE2 with TAK-981 results in significant MCL cell death in vitro and in vivo with mitotic dysregulation being an important mechanism of action. We uncovered a sumoylation program in mitotic MCL cells comprised of multiple pathways which could be directly targeted with TAK-981. Centromeric localization of topoisomerase 2A, a gene highly upregulated in SAE1 and SAE2 overexpressing MCL cells, was lost with TAK-981 treatment likely contributing to the mitotic dysregulation seen in MCL cells. CONCLUSIONS: This study not only validates SAE2 as a therapeutic target in MCL but also opens the door to further mechanistic work to uncover how to best use desumoylation therapy to treat MCL and other lymphoid malignancies.

Whole-Genome Sequence of Pantoea americana Strain VS1, an Extended-Spectrum ß-Lactamase-Producing Epibiont Isolated from Magnolia grandiflora.

Pantoea americana strain VS1, an extended-spectrum ß-lactamase-producing epibiont, was isolated from Magnolia grandiflora in central Florida, USA. Here, we report the de novo whole-genome sequence of this strain, which consists of a total of 191 contigs spanning 5,412,831 bp, with a GC content of 57.3% and comprising 4,836 predicted coding sequences.

De Novo Whole-Genome Sequence of Pantoea latae Strain AS1, Isolated from Zamia floridana Rhizosphere in Central Florida, USA.

Pantoea latae strain AS1 was isolated from the rhizophere of a cycad, Zamia floridana, in central Florida, USA. Here, we report the de novo whole-genome sequence of this strain, which consists of a total of 83 contigs spanning 4,960,415 bp, with a G+C content of 59.6%, and comprising 4,527 predicted coding sequences.

Whole-Genome Sequence of Bacillus stratosphericus Strain 5Co, Isolated from Lichen Usnea florida in Central Florida, United States, with High Tolerance to Salt and Heavy Metal.

Bacillus stratosphericus strain 5Co was isolated from lichen Usnea florida in central Florida, United States. Here, we report a draft genome sequence of this strain, which consists of 159 contigs spanning 3,628,496 bp, with a G+C content of 41.3% and comprises 3,729 predicted coding sequences.

Draft Genome Sequences of Extended-Spectrum-Beta-Lactamase-Producing Morganella morganii Strains AA1 and AV1, Isolated from a Freshwater Lake and Eicchorniacrassipes Roots.

Two strains of Morganella morganii, AA1 and AV1, were isolated from freshwater and Eicchornia crassipes roots, respectively. Here, we report their draft genome sequences, which are ~3.6 Mb and have 51% G+C content. The predicted coding sequences (3,259 for strain AA1 and 3,345 for strain AV1) encode beta-lactamases, transpeptidases, and penicillin-binding proteins.

Deep Sequencing-Identified Kanamycin-Resistant Paenibacillus sp. Strain KS1 Isolated from Epiphyte Tillandsia usneoides (Spanish Moss) in Central Florida, USA.

Paenibacillus sp. strain KS1 was isolated from an epiphyte, Tillandsia usneoides (Spanish moss), in central Florida, USA. Here, we report a draft genome sequence of this strain, which consists of a total of 398 contigs spanning 6,508,195 bp, with a G+C content of 46.5% and comprising 5,401 predicted coding sequences.

Multiplex PCR based genotypic characterization of pathogenic vancomycin resistant Enterococcus faecalis recovered from an Indian river along a city landscape.

BACKGROUND: Enterococci are normal commensals of human gut, but vancomycin-resistant enterococci (VRE) are a severe threat to human health. Antimicrobial-resistant enterococci have been reported previously from Indian surface waters. However, the presence of antimicrobial resistance and virulence markers in Enterococcus faecalis, the most dominant enterococci is yet to be investigated. OBJECTIVES: The goal of this study was to analyse concentration of enterococci and distribution of antimicrobial resistance and virulence markers in E. faecalis isolates from river waters along an important north Indian city landscape. METHODS: We enumerated enterococci in river water samples (n = 60) collected from five sites across the Lucknow city landscape using the most probable number and membrane-filtration methods. The antimicrobial sensitivity profile of E. faecalis isolate was generated with the Kirby-Bauer antimicrobial disc diffusion assay. The multiplex PCR was used for genotypic characterization of vancomycin-resistance and virulence in E. faecalis isolates. RESULTS: Enterococci density (p < 0.0001) increased from up-to-down-stream sites. Multiplex PCR based genotypic characterization has shown a significant distribution of virulence-markers gelE, ace or efaA in the E. faecalis isolates (p < 0.05). The range of antimicrobial-resistance varied from 5 to 12 in the landscape with the frequency of vancomycin-resistant E. faecalis (VRE) ranging from 22 to 100 %. CONCLUSION: The occurrence of pathogenic VRE in river Gomti surface water is an important health concern. The observed high background pool of resistance and virulence in E. faecalis in river waters has the potential to disseminate more alarming antimicrobial resistance in the environment and poses serious health risk in developing countries like India as VRE infections could lead to increased cost of healthcare.

HO-3867, a safe STAT3 inhibitor, is selectively cytotoxic to ovarian cancer.

STAT3 is well corroborated preclinically as a cancer therapeutic target, but tractable translational strategies for its blockade by small molecule inhibitors have remained elusive. In this study, we report the development of a novel class of bifunctional STAT3 inhibitors, based on conjugation of a diarylidenyl-piperidone (DAP) backbone to an N-hydroxypyrroline (-NOH) group, which exhibits minimal toxicity against normal cells and good oral bioavailability. Molecular modeling studies of this class suggested direct interaction with the STAT3 DNA binding domain. In particular, the DAP compound HO-3867 selectively inhibited STAT3 phosphorylation, transcription, and DNA binding without affecting the expression of other active STATs. HO-3867 exhibited minimal toxicity toward noncancerous cells and tissues but induced apoptosis in ovarian cancer cells. Pharmacologic analysis revealed greater bioabsorption and bioavailability of the active (cytotoxic) metabolites in cancer cells compared with normal cells. The selective cytotoxicity of HO-3867 seemed to be multifaceted, eliciting differential activation of the Akt pathway in normal versus cancer cells. RNAi attenuation experiments confirmed the requirement of STAT3 for HO-3867-mediated apoptosis in ovarian cancer cells. In vivo testing showed that HO-3867 could block xenograft tumor growth without toxic side effects. Furthermore, in primary human ovarian cancer cells isolated from patient ascites, HO-3867 inhibited cell migration/invasion and survival. Our results offer preclinical proof-of-concept for HO-3867 as a selective STAT3 inhibitor to treat ovarian cancer and other solid tumors where STAT3 is widely upregulated.

Expression profiling of toxicity pathway genes by real-time PCR array in cypermethrin-exposed mouse brain.

Cypermethrin, a type II pyrethroid, has been shown to exert genotoxic effects in the central nervous system of non-target species such as mouse and Drosophila. To unravel the gene expression of toxicity-related pathways in cypermethrin-exposed Swiss albino mouse brain, transcriptional profiling was carried out through pathway-focused real-time PCR arrays (DNA damage signaling, oxidative stress/antioxidants, and stress/toxicity pathways). The real-time PCR array data revealed a significant (p < 0.05) modulation in transcript levels of 61 genes involved in DNA replication and repair, apoptosis, cell cycle, oxidative stress, and toxicity pathways. Cypermethrin also produced oxidative stress in brain, as was evident by a significant (p < 0.05) elevation (66%) in lipid peroxidation and reduction of glutathione (GSH) content (10.6%) as well as catalase activity (56.7%). The results demonstrate that cypermethrin alters the expression of stress- and toxicity-related genes as well as induces oxidative stress which may lead to DNA damage. These observations also point to complex metabolic networks involved in genotoxic manifestations by cypermethrin.

Enterococci in river Ganga surface waters: propensity of species distribution, dissemination of antimicrobial-resistance and virulence-markers among species along landscape.

BACKGROUND: Surface waters quality has declined in developing countries due to rapid industrialization and population growth. The microbiological quality of river Ganga, a life-sustaining surface water resource for large population of northern India, is adversely affected by several point and non-point sources of pollution. Further, untreated surface waters are consumed for drinking and various household tasks in India making the public vulnerable to water-borne diseases and outbreaks. Enterococci, the 'indicator' of water quality, correlates best with the incidence of gastrointestinal diseases as well as prevalence of other pathogenic microorganisms. Therefore, this study aims to determine the distribution of species diversity, dissemination of antimicrobial-resistance and virulence-markers in enterococci with respect to rural-urban landscape along river Ganga in northern India. RESULTS: Enterococci density (chi2: 1900, df: 1; p < 0.0001) increased from up-to-down gradient sites in the landscape. Species diversity exhibit significant (chi2: 100.4, df: 20; p < 0.0001) and progressive distribution of E. faecalis, E. faecium, E. durans and E. hirae down the gradient. Statistically discernible (p: 0.0156 - < 0.0001) background pool of resistance and virulence was observed among different Enterococcus spp. recovered from five sites in the up-to-down gradient landscape. A significant correlation was observed in the distribution of multiple-antimicrobial-resistance (viz., erythromycin-rifampicin-gentamicin-methicillin and vancomycin-gentamicin-streptomycin; rs: 0.9747; p: 0.0083) and multiple-virulence-markers (viz., gelE+esp+; rs: 0.9747; p: 0.0083; gelE+efaA+; rs: 0.8944; p: 0.0417) among different Enterococcus spp. CONCLUSION: Our observations show prevalence of multiple-antimicrobial-resistance as well as multiple-virulence traits among different Enterococcus spp. The observed high background pool of resistance and virulence in enterococci in river waters of populous countries has the potential to disseminate more alarming antimicrobial-resistant pathogenic bacteria of same or other lineage in the environment. Therefore, the presence of elevated levels of virulent enterococci with emerging vancomycin resistance in surface waters poses serious health risk in developing countries like India.

Real time PCR for the rapid detection of vanA gene in surface waters and aquatic macrophyte by molecular beacon probe.

Enterococci serve as an "indicator" of fecal contamination for recreational water quality. The vancomycin-resistant-enterococci (VRE) are emerging environmental contaminants in the surface waters. The aim ofthis study wasto develop a rapid and specific molecular beacon probe (MBP)-based real-time PCR assay for detection of vanA gene in surface waters and aquatic macrophyte. The limit of detection (LOD) of the MBP assay was 1 CFU/mL of VRE [r = 0.943; PCR efficiency = 99.7%] in 2-fold dilution format within 2.5 h and demonstrated high specificityfor environmental enterococci isolates exhibiting VanA phenotype (n=25). VRE were detected from downstream surface waters of the rivers impacted by point sources of pollution and recreational activities.The probe detected vanA gene in rootmat associated microbiota of E. crassipes (Mart) Solms. an aquatic nuisance weed, at eutrophic sites of the surface waters (ANOVA p < 0.001). In addition, the assay enabled detection of otherwise nondetectable vanA gene concentration in the upstream sites of two Indian rivers (Student's ttest p < 0.001). The MBP assay developed can be used for sensitive and rapid detection of VRE in surface waters and identification of nonpoint sources of pollution for implementation of preventive measures to protect human health.