Normal Life Cycle Research Articles

Model-Driven Availability Assessment of the NFV-MANO With Software Rejuvenation

Network Function Virtualization enables network operators to modernize their networks with greater elasticity, network programmability, and scalability. Exploiting these advantages requires new and specialized designs for management, automation, and orchestration systems which are capable of reliably operating and handling new elements such as virtual functions, virtualized infrastructures, and a whole new set of relationships among them. Operations such as resource allocation, instantiation, monitoring, scaling, or termination of virtual functions are key lifecycle operations that NFV management and orchestration (NFV-MANO) frameworks need to correctly perform. Failures of the NFV-MANO prevent the network ability to respond to new service requests or events related to the normal lifecycle operation of network services. Thus, it is important to ensure robustness and high availability of the MANO framework. This paper adopts a model-driven approach to predict the availability of the NFV-MANO and assess the impact that different failure modes have. We propose different models, based on Stochastic Activity Networks (SANs), which abstract various MANO deployment configurations, inspired by current containerized open-source MANO implementations. Moreover, we integrate software rejuvenation and investigate the trade-off between its associated overhead and system availability increase. An extensive experimental campaign with fault-injection techniques on a real-life MANO implementation allows to derive a number of realistic recovery parameters. The case studies are used to quantitatively evaluate the steady-state availability and identify the most important parameters influencing system availability for the different deployment configurations.

UHPLC-IM-Q-ToFMS analysis of maradolipids, found exclusively in Caenorhabditis elegans dauer larvae

Lipid identification is one of the current bottlenecks in lipidomics and lipid profiling, especially for novel lipid classes, and requires multidimensional data for correct annotation. We used the combination of chromatographic and ion mobility separation together with data-independent acquisition (DIA) of tandem mass spectrometric data for the analysis of lipids in the biomedical model organism Caenorhabditis elegans. C. elegans reacts to harsh environmental conditions by interrupting its normal life cycle and entering an alternative developmental stage called dauer stage. Dauer larvae show distinct changes in metabolism and morphology to survive unfavorable environmental conditions and are able to survive for a long time without feeding. Only at this developmental stage, dauer larvae produce a specific class of glycolipids called maradolipids. We performed an analysis of maradolipids using ultrahigh performance liquid chromatography-ion mobility spectrometry-quadrupole-time of flight-mass spectrometry (UHPLC-IM-Q-ToFMS) using drift tube ion mobility to showcase how the integration of retention times, collisional cross sections, and DIA fragmentation data can be used for lipid identification. The obtained results show that combination of UHPLC and IM separation together with DIA represents a valuable tool for initial lipid identification. Using this analytical tool, a total of 45 marado- and lysomaradolipids have been putatively identified and 10 confirmed by authentic standards directly from C. elegans dauer larvae lipid extracts without the further need for further purification of glycolipids. Furthermore, we putatively identified two isomers of a lysomaradolipid not known so far.Graphical abstract

EXPERIENCE IN THE SUCCESSFUL MANAGEMENT OF EIGHT CASES OF CONGENITAL VAGINAL SEPTA

Background: Vaginal septum is a rare congenital abnormality of the female genital tract. It can be asymptomatic or may be present with a variety of symptoms like amenorrhea, dyspareunia, dysmenorrhea, infertility and abortions. Aims And Objectives: To study patients with vaginal septum presenting at our hospital in detail with clinical ndings , investigations and their surgical management. Materials And Methods: A prospective study of eight cases of vaginal septum diagnosed at ASRAM Medical College ,Eluru. Results: All cases of transverse and longitudinal vaginal septum were successfully operated, had normal sexual life and menstrual cycles. They conceived and had children varying from one to three.

CDNA-Derived RNA Phage Assembly Reveals Critical Residues in the Maturation Protein of the Pseudomonas aeruginosa Leviphage PP7.

PP7 is a leviphage, with a single-stranded RNA genome, that infects Pseudomonas aeruginosa PAO1. A reverse genetic system for PP7 was previously created by using reverse-transcribed cDNA (PP7O) from a virion-derived RNA genome. Here, we have found that the PP7O cDNA contained 20 nucleotide differences from the PP7 genome sequence deposited in the database. We created another reverse genetic system exploiting chemically synthesized cDNA (PP7S) based on the database sequence. Unlike PP7O, which yielded infectious PP7 virions, PP7S-derived particles were incapable of plaque formation on PAO1 cells, which was restored in the PAO1 cells expressing the maturation protein (MP) from PP7O Using this reverse genetic system, we revealed two amino acid residues involved in the known roles of MP (i.e., adsorption and genome replication), fortuitously providing a lesson that the viral RNA genome sequencing needs functional verification, possibly by a reverse genetic system.IMPORTANCE The biological significance of RNA phages has been largely ignored, ironically, because few studies have focused on RNA phages. As an initial attempt to properly represent RNA phages in the phageome, we previously created, by using reverse-transcribed cDNA, a reverse genetic system for the small RNA phage PP7, which infects the opportunistic human pathogen Pseudomonas aeruginosa We report another system by using chemically synthesized cDNA based on the database genome that has 20 nucleotide differences from the previous cDNA. Investigation of those cDNA-derived phage virions revealed that two amino acids of the maturation protein are crucial for the normal phage lifecycle at different steps. Our study provides insight into the molecular basis for the RNA phage lifecycle and a lesson that the RNA genome sequencing needs to be carefully validated by cDNA-based phage assembly systems.

Suppression of human papillomavirus type 16 E5 oncoprotein: A promising step in fostering the treatment of cervical cancer.

Cervical cancer is a growing global disease in developing countries. Persistent infection with human papillomaviruses (HPV) is an essential causative agent in this type of cancer. Several studies demonstrate HPV E5 oncoprotein can impress the normal life cycle of HPV-infected cells by targeting some pivotal cellular signaling pathways, such as the epidermal growth factor receptor (EGFR) signaling pathway. In this study, we used E5-siRNA to knockdown that essential oncogene and considered the effect of E5 silencing on proliferation, apoptosis, cell cycle, apoptosis-related gene expression, and the initiator of the EGFR signaling pathway in cervical cancer cells. The results demonstrate that E5 plays an essential role in the proliferation and inhibited apoptosis in cervical cancer. Furthermore, silencing E5 reduces proliferation, increases apoptosis, and elevates related-genes expression of these malignant cells. Overall, E5 suppression may be appropriate for ameliorating cervical cancer progression.

Membrane Processing and Steady-State Regulation of the Alternative Peroxisomal Import Receptor Pex9p.

Import of peroxisomal matrix proteins with a type 1 peroxisomal targeting signal (PTS1) in Saccharomyces cerevisiae is facilitated by cytosolic import receptors Pex5p and Pex9p. While Pex5p has a broad specificity for all PTS1 proteins independent of the growth conditions, Pex9p is only expressed in fatty-acid containing media to mediate peroxisomal import of the two malate synthases, Mls1p and Mls2p, as well as the glutathione transferase Gto1p. Pex5p-cargo complexes dock at the peroxisomal membrane, translocate their cargo-protein via a transient pore and are recycled into the cytosol for a further round of import. The processing of Pex5p has been shown to require a complex network of interactions with other membrane-bound peroxins, as well as decoration with ubiquitin as signal for its ATP-dependent release and recycling. Here, we show that the alternative receptor Pex9p requires the same set of interacting peroxins to mediate peroxisomal import of Mls1p. However, while Pex5p is rather stable, Pex9p is rapidly degraded during its normal life cycle. The steady-state regulation of Pex9p, combining oleate-induced expression with high turnover rates resembles that of Pex18p, one of the two co-receptors of the PTS2-dependent targeting pathway into peroxisomes. Both Pex9p- and Pex18p-dependent import routes serve the fast metabolic adaptation to changes of carbon sources in baker’s yeast. By sequence similarities, we identified another Pex9p homolog in the human pathogenic fungus Candida glabrata, in which similar metabolic reprogramming strategies are crucial for survival of the pathogen.

Sequencing [3+2]-cycloaddition and multicomponent reactions: A regioselective microwave-assisted synthesis of 1,4-disubstituted 1,2,3-triazoles using ionic liquid supported Cu(II) precatalysts in methanol

Heterocyclic compounds with two to three nitrogen atoms play a pivotal role in the normal life cycle of a cell. Further the design and synthesis of a quality heterocyclic compound library with N-atoms as new chemical probes active, is vital in drug discovery. In this context, an efficient one-pot multicomponent strategy for the synthesis of a mini library of 1,4-disubstituted 1,2,3-triazoles is described. This new multicomponent one-pot method features a combination of ionic liquid supported Cu(II) precatalysts in methanol catalyzed [3+2]-cycloaddition with microwave irradiation reactions. The synthetic manipulation involved the efficient reduction of ionic liquid supported Cu(II) catalyst by methanol followed by [3+2] cycloaddition with alkynes using in situ generated azides to obtain 1,2,3-triazoles regioselectively.

Central role of 70-kDa heat shock protein in adaptation of plants to drought stress

The 70-kDa heat shock proteins (HSP70s) are a conserved class of chaperones that play critical roles during the normal life cycle of plants. HSP70s are particularly involved in the regulation of biotic and abiotic stress responses. In this paper, the potential roles of this protein were investigated. A reverse genetic approach was employed for transient silencing of hsp70 gene in tomato (Solanum lycopersicum L.) to evaluate different growth and physiological parameters under normal conditions and during the response to drought stress. A combined ANOVA (analysis of variance) and HCA (hierarchical clustering analysis) showed that hsp70 silencing led to severe growth retardation and mortality, significant membrane damage and leakage, decline in relative water content, low rate of pigment accumulation, and reduced antioxidant enzyme activity under normal and drought stress conditions. Among the different parameters, proline was the only trait that was unaffected by gene silencing and accumulated by similar amounts to that of nonsilent plants. In conclusion, HSP70 played critical roles in maintaining the cellular homeostasis of plants during adaptation to drought and under normal plant life conditions. It was speculated that proline was, to some extent, involved in improving the loss of protein folding or function resulting from HSP70 deficiency, and played a crucial role in the adaptation of plants on exposure to stress.

GFP transient expression and silencing in Fragaria x ananassa

BACKGROUND: Stable transformation, transient expression, and post-transcriptional gene silencing (PTGS) are powerful methodologies that allow exploration of gene function. OBJECTIVE: We aimed to apply these methodologies to strawberry leaves. Methods: the binary vectors pBIN19-sgfp, pBICdsGFP and pBIN61-P19 were transferred into A. tumefaciens EHA105 supervirulent strain by electroporation. The sgfp gene silencing was carried out in stably transformed GFP (green fluorescent protein) F. x ananassa Duch. cultivar ‘Pájaro’ strawberry plants by agroinfiltration. GFP-fluorescence was observed using a stereomicroscope (507 nm). RESULTS: We attained a GFP transgenic F. x ananassa plant that expresses the functional protein in all the tissues during a complete and normal life cycle. In planta sgfp transient expression and silencing have also been achieved in F. x ananassa cv. ‘Pájaro’ leaves of wild type and GFP transgenic plants, respectively. Agrobacterium-mediated transient expression was visualized as high intensity green fluorescent spots as early as 7 days post-agroinfiltration (dpa), peaking between 10 and 14 dpa and persisting as long as 24 dpa. A knockdown GFP phenotype was achieved by silencing using a dsGFP hairpin. CONCLUSION: This work contributes significantly to the reverse genetics field in strawberry, might help to gain knowledge in the analysis of functional promoters and thereby allow protein expression and silencing of genes. This will help to develop resistant plants expressing plant defense elicitors or silencing pathogen receptors and/or negative regulators of plant defense.

Biomarker screening for the early detection of prostate cancer using an exosomal-enrichment RNA liquid biopsy test.

e15553 Background: Prostate cancer is one of the most common cancers in men, with approximately 10% of all new cancer cases and ~5% of all cancer deaths in 2019. The standard test for prostate cancer is the Prostate Serum Antigen (PSA) test. The PSA test suffers from low specificity (20-40%) in patients including ‘grey zone’ levels (4-10 ng/mL); moreover, the PSA test fails to identify patients with high-risk cancers. Previously we developed ExoDx Prostate Intelliscore (EPI), a urine-based exosome prostate cancer test optimized to rule out the need for a biopsy (risk stratification for high-grade prostate cancer). This study utilized a next generation exosome-based test that specifically enriches a subtype of prostate cancer exosomes from urine. Early detection of prostate cancer via a non-invasive method is desirable and the identification of patients with high-risk cancer is critical. Here we describe the development of a prostate-specific urinary exosome test for the identification of patients with prostate cancer. Methods: We have developed a prostate-specific enrichment method to isolate exosomes of prostate origin from urine samples. Using an affinity-based method against surface marker proteins found on prostate cells, we were able to selectively enrich for exosomes shed by the prostate gland with demonstrated specificity. Subsequent analysis of exosomal nucleic acids enables a promising panel of gene expression biomarkers capable of distinguishing patients with prostate cancer from healthy individuals. Results: RNA from prostate cancer enriched exosomes was compared to total exosomes from urine. Enrichment of prostate cancer specific exosomes significantly enhanced the RNA signature compared to total urine exosomes. Conclusions: Prostate cancer tests have recently been developed for RNA signatures in urine. Exosomes provide a source of nucleic acids as they are actively shed continuously from living cells as part of their normal life cycle. The urine exosomes can be used for total RNA transcriptome analysis and are therefore a very rich source of biomarkers for prostate cancer that can be tailored to different clinical indications. An affinity-based enrichment for tissue-specific exosomes allow for better resolution of the gene expression profile from the tissue of interest and reduces the RNA targets from non-relevant processes of the bladder and kidneys. The gene signature identified in this ongoing study could potentially provide a non-invasive molecular means for the early diagnosis of prostate cancer.

An assessment study of known pyrazolopyrimidines: Chemical methodology and cellular activity.

Heterocyclic compounds with nitrogen atom play a key role in the normal life cycle of a cell. Pyrazolopyrimidine is a privileged class of nitrogen containing fused heterocyclic compound contributing to a major portion of all lead molecules in medicinal chemistry. The thumbprint of pyrazolopyrimidine as a pharmacophore is always noticeable due to its analogy with the adenine base in DNA. Pyrazolopyrimidines are divided into five types [I, II, III, IV, V] based on the mechanism of action on the specific target conferring a wide scope of research which has accelerated the interest of researchers to investigate its biological profile. In 1956, the anti-cancer activity of pyrazolopyrimidine was evaluated for the first time with appreciable results. Since then, medicinal chemists centered their work on various methods of synthesis and evaluating the biological profile of pyrazolopyrimidine isomers. This report consists of novel methodologies followed to synthesize pyrazolopyrimidine isomers along with a note on their biological significance. To the best of our knowledge, this review article will be first of its kind to encompass different synthetic procedures along with anti-cancer, kinase inhibition, phosphodiesterase inhibition and receptor blocking activity of pyrazolopyrimidine moieties. IC50 values of potent compounds are added wherever necessary to understand the suitability of pyrazolopyrimidine skeletons for a specific biological activity.

HPV Oncoproteins and the Ubiquitin Proteasome System: A Signature of Malignancy?

Human papillomavirus (HPV) E6 and E7 oncoproteins are critical for development and maintenance of the malignant phenotype in HPV-induced cancers. These two viral oncoproteins interfere with a plethora of cellular pathways, including the regulation of cell cycle and the control of apoptosis, which are critical in maintaining normal cellular functions. E6 and E7 bind directly with certain components of the Ubiquitin Proteasome System (UPS), enabling them to manipulate a number of important cellular pathways. These activities are the means by which HPV establishes an environment supporting the normal viral life cycle, however in some instances they can also lead to the development of malignancy. In this review, we have discussed how E6 and E7 oncoproteins from alpha and beta HPV types interact with the components of the UPS, and how this interplay contributes to the development of cancer.

Effect of Lipid Structure and Material Properties on the Membrane Stability to Pore Formation

Biological membranes are natural barriers that preserve the integrity of the cell and its organelles. Even temporary damage of the cell membrane can break its normal homeostasis that will start the recovery mechanisms, apoptosis and inflammatory reactions. Formation of pores, through defects, in the lipid bilayer of biological membranes can lead to serious disruption of the normal lifecycle of the cells and the development of pathologies.A two-pronged approach was used to study the energetics of pore formation in lipid membranes assembled by isoprenoid lipids (e.g., DPhPC and Di-O-PhPC). The first prong studied the line tension of DPhPC, Di-O-PhPC, DOPC, and POPC using MD simulations and experiments with an electrical breakdown technique. Resistance to pore formation can be characterized by pore edge line tension, which describes the energy barrier height that should be overcome to irreversibly expand the pore, i.e. to rupture the membrane. A method of measuring pore line tension in MD, determining the lateral tension of the lipid membrane with the static pore, and experiments revealed that branched lipid significantly increased the pore energy and membrane resistance to pore formation. Additionally, work on GUVs and MD showed that low-weight solvent significantly lowers pore edge energy. The second prong used MD to quantify other key material properties that can affect pore formation (i.e., the compressibility modulus, bending modulus, per area free energy change with respect to curvature, and the intrinsic curvature). DPhPC's bending modulus is about half that of DPPC, but DPhPC's compressibility modulus is about two times larger than DPPC. Finally, DPhPC's effect on monolayer spontaneous curvature is similar to that of a PE lipid. Thus both structural peculiarities of lipid tails and solvents strongly influence membrane stability.

IPR CELEIRO: Common bean cultivar moderately resistant to bean golden mosaic virus

Abstract IPR Celeiro is a common bean (Phaseolus vulgaris L.) cultivar with a carioca-type seed and moderate resistance to Bean Golden Mosaic Virus (BGMV). IPR Celeiro has a normal life cycle and an upright architecture. This cultivar was released for the purpose of integrating its use with disease management systems in the cultivation areas affected by this virus.

The effects of Centella asiatica (L.) Urban on neural differentiation of human mesenchymal stem cells in vitro

BackgroundCentella asiatica (L.) Urban, known as Indian Pennywort, is a tropical medicinal plant from Apiaceae family native to Southeast Asian countries. It has been widely used as a nerve tonic in Ayuverdic medicine since ancient times. However, whether it can substitute for neurotrophic factors to induce human mesenchymal stem cell (hMSCs) differentiation into the neural lineage remains unknown. This study aimed to investigate the effect of a raw extract of C. asiatica (L.) (RECA) on the neural differentiation of hMSCs in vitro.MethodsThe hMSCs derived from human Wharton’s jelly umbilical cord (hWJMSCs; n = 6) were treated with RECA at different concentrations; 400, 800, 1200, 1600, 2000 and 2400 μg/ml. The cytotoxicity of RECA was evaluated via the MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) and cell proliferation assays. The hWJMSCs were then induced to neural lineage for 9 days either with RECA alone or RECA in combination with neurotrophic factors (NF). Cell morphological changes were observed under an inverted microscope, while the expression of the neural markers S100β, p75 NGFR, MBP, GFAP and MOG was analyzed by quantitative polymerase chain reaction and immunocytochemistry. The cell cycle profile of differentiated and undifferentiated hWJMSCs was investigated through cell cycle analysis.ResultsRECA exerted effects on both proliferation and neural differentiation of hWJMSCs in a dose-dependent manner. RECA reduced the proliferation of hWJMSCs and was cytotoxic to cells above 1600 μg/ml, with IC50 value, 1875 ± 55.67 μg/ml. In parallel with the reduction in cell viability, cell enlargement was also observed at the end of the induction. Cells treated with RECA alone had more obvious protein expression of the neural markers compared to the other groups. Meanwhile, gene expression of the aforementioned markers was detected at low levels across the experimental groups. The supplementation of hWJMSCs with RECA did not change the normal life cycle of the cells.ConclusionsAlthough RECA reduced the proliferation of hWJMSCs, a low dose of RECA (400 μg/ml), alone or in combination of neurotrophic factors (NF + RECA 400 μg/ml), has the potential to differentiate hWJMSCs into Schwann cells and other neural lineage cells.

Temporary Solubilizing Tags Method for the Chemical Synthesis of Hydrophobic Proteins

Hydrophobic proteins, as one of the cellular protein classifications, play an essential function in maintaining the normal life cycle of living cells. Researches on the structure and function of hydrophobic proteins promote the exploration of the causes of major diseases, and development of new therapeutic agents for disease treatment. However, the poor water solubility of hydrophobic proteins creates problems for their preparation, separation, characterization and functional studies. The temporary solubilizing tags are considered a practical strategy to effectively solve the poor water solubility problem of hydrophobic proteins. This strategy can significantly improve the water solubility of hydrophobic peptides/proteins, making them like water-soluble peptides/proteins easy to be purified, characterized. More importantly, the temporary solubilizing tags can be removed after protein synthesis, so thus the structure and function of the hydrophobic proteins are not affected. At present, temporary solubilizing tags have been successfully used to prepare many important hydrophobic proteins such as membrane proteins, lipoproteins and chaperones. In this review, we summarize the recent researches and applications of temporary solubilizing tags.

A novel triple amino acid substitution in the EPSPS found in a high-level glyphosate-resistant Amaranthus hybridus population from Argentina.

The evolution of herbicide-resistant weeds is one of the most important concerns of global agriculture. Amaranthus hybridus L. is a competitive weed for summer crops in South America. In this article, we intend to unravel the molecular mechanisms by which an A. hybridus population from Argentina has become resistant to extraordinarily high levels of glyphosate. The glyphosate-resistant population (A) exhibited particularly high parameters of resistance (GR50 = 20 900 g ai ha-1 , Rf = 314), with all plants completing a normal life cycle even after 32X dose application. No shikimic acid accumulation was detected in the resistant plants at any of the glyphosate concentrations tested. Molecular and genetic analyses revealed a novel triple substitution (TAP-IVS: T102I, A103V, and P106S) in the 5-enol-pyruvylshikimate-3-phosphate synthase (EPSPS) enzyme of population A and an incipient increase on the epsps relative copy number but without effects on the epsps transcription levels. The novel mechanism was prevalent, with 48% and 52% of the individuals being homozygous and heterozygous for the triple substitution, respectively. In silico conformational studies revealed that TAP-IVS triple substitution would generate an EPSPS with a functional active site but with an increased restriction to glyphosate binding. The prevalence of the TAP-IVS triple substitution as the sole mechanism detected in the highly glyphosate resistant population suggests the evolution of a new glyphosate resistance mechanism arising in A. hybridus. This is the first report of a naturally occurring EPSPS triple substitution and the first glyphosate target-site resistance mechanism described in A. hybridus. © 2018 Society of Chemical Industry.

A small cysteine-rich protein from two kingdoms of microbes is recognized as a novel pathogen-associated molecular pattern.

Pathogen-associated molecular patterns (PAMPs) are conserved molecules that are crucial for normal life cycle of microorganisms. However, the diversity of microbial PAMPs is little known. During screening of cell-death-inducing factors from the necrotrophic fungus Valsa mali, we identified a novel PAMP VmE02 that is widely spread in oomycetes and fungi. Agrobacterium tumefaciens-mediated transient expression or infiltration of recombinant protein produced by Escherichia coli was performed to assay elicitor activity of the proteins tested. Virus-induced gene silencing in Nicotiana benthamiana was used to determine the components involved in VmE02-triggered cell death. The role of VmE02 in virulence and conidiation of V.mali were characterized by gene deletion and complementation. We found that VmE02, together with some of its homologues from both oomycete and fungal species, exhibited cell-death-inducing activity in N.benthamiana. VmE02-triggered cell death was shown to be dependent on BRI1-ASSOCIATED KINASE-1, SUPPRESSOR OF BIR1-1, HSP90 and SGT1 in N.benthamiana. Deletion of VmE02 in V.mali greatly attenuated pathogen conidiation but not virulence, and treatment of N.benthamiana with VmE02 enhances plant resistance to Sclerotinia sclerotiorum and Phytophthora capsici. We conclude that VmE02 is a novel cross-kingdom PAMP produced by several fungi and oomycetes.

Inhibition of Epstein-Barr Virus Replication in Human Papillomavirus-Immortalized Keratinocytes

Epstein-Barr virus (EBV) is implicated in the pathogenesis of human papillomavirus (HPV)-associated oropharyngeal squamous cell carcinoma (OSCC). EBV-associated cancers harbor a latent EBV infection characterized by a lack of viral replication and the expression of viral oncogenes. Cellular changes promoted by HPV are comparable to those shown to facilitate EBV latency, though whether HPV-positive cells support a latent EBV infection has not been demonstrated. Using a model of direct EBV infection into HPV16-immortalized tonsillar cells grown in organotypic raft culture, we showed robust EBV replication in HPV-negative rafts but little to no replication in HPV-immortalized rafts. The reduced EBV replication was independent of immortalization, as human telomerase-immortalized normal oral keratinocytes supported robust EBV replication. Furthermore, we observed reduced EBV lytic gene expression and increased expression of EBER1, a noncoding RNA highly expressed in latently infected cells, in the presence of HPV. The use of human foreskin keratinocyte rafts expressing the HPV16 E6 and/or E7 oncogene(s) (HPV E6 and E7 rafts) showed that E7 was sufficient to reduce EBV replication. EBV replication is dependent upon epithelial differentiation and the differentiation-dependent expression of the transcription factors KLF4 and PRDM1. While KLF4 and PRDM1 levels were unaltered, the expression levels of KLF4 transcriptional targets, including late differentiation markers, were reduced in HPV E6 and E7 rafts compared to their levels in parental rafts. However, the HPV E7-mediated block in EBV replication correlated with delayed expression of early differentiation markers. Overall, this study reveals an HPV16-mediated block in EBV replication, through E7, that may facilitate EBV latency and long-term persistence in the tumor context.IMPORTANCE Using a model examining the establishment of EBV infection in HPV-immortalized tissues, we showed an HPV-induced interruption of the normal EBV life cycle reminiscent of a latent EBV infection. Our data support the notion that a persistent EBV epithelial infection depends upon preexisting cellular alterations and suggest the ability of HPV to promote such changes. More importantly, these findings introduce a model for how EBV coinfection may influence HPV-positive (HPV-pos) OSCC pathogenesis. Latently EBV-infected epithelial cells, as well as other EBV-associated head-and-neck carcinomas, exhibit oncogenic phenotypes commonly seen in HPV-pos OSCC. Therefore, an HPV-induced shift in the EBV life cycle toward latency would not only facilitate EBV persistence but also provide additional viral oncogene expression, which can contribute to the rapid progression of HPV-pos OSCC. These findings provide a step toward defining a role for EBV as a cofactor in HPV-positive oropharyngeal tumors.

Strongyloides stercoralis larvae in the urine of a patient with transitional cell carcinoma of the bladder: a case report.

Disseminated infection with Strongyloides stercoralis refers to the massive migration of infective larvae from the gastrointestinal tract to other organs that are not involved in the normal life cycle of the parasite. We describe the case of a Nigerian male with transitional cell carcinoma of the bladder in whom larvae of S. stercoralis was identified in the urine. This report involves a 60-year old male Nigerian presenting to the Urology clinic of the Jos University teaching hospital, Nigeria with disseminated S. stercoralis. The index patient presented with a 5month history of total haematuria, urinary frequency, urgency, nocturia, straining to pass urine, feeling of incomplete voiding and terminal dribbling. He also had episodes of suprapubic pain. Physical examination revealed a cachexic patient who had mild suprapubic tenderness. Urinary examination showed numerous red blood cells and rhabditiform larvae of S. stercoralis. Abdominal ultrasound revealed a heterogeneous mass in the urinary bladder measuring 4.0 × 3.3cm. Abdominal computed tomography also showed an irregular mass measuring 4.2 × 3.8cm with HU of 41 projecting into the bladder from the posterior wall towards the dome. Histology of the biopsy specimen revealed transitional cell carcinoma. The patient was treated with a single dose of oral ivermectin but died 1 week later. Physicians working in areas that are endemic for S. stercoralis should consider investigating immunocompromised patients for S. stercoralis infection given the poor prognosis of disseminated infection in this group of patients.