Sustained proliferation in cancer: mechanisms and novel therapeutic targets

Proliferation is an important part of cancer development and progression. This is manifest by altered expression and/or activity of cell cycle related proteins. Constitutive activation of many signal transduction pathways also stimulates cell growth. Early steps in tumor development are associated with a fibrogenic response and the development of a hypoxic environment which favors the survival and proliferation of cancer stem cells. Part of the survival strategy of cancer stem cells may manifested by alterations in cell metabolism. Once tumors appear, growth and metastasis may be supported by overproduction of appropriate hormones (in hormonally dependent cancers), by promoting angiogenesis, by undergoing epithelial to mesenchymal transition, by triggering autophagy, and by taking cues from surrounding stromal cells. A number of natural compounds (e.g., curcumin, resveratrol, indole-3-carbinol, brassinin, sulforaphane, epigallocatechin-3-gallate, genistein, ellagitannins, lycopene and quercetin) have been found to inhibit one or more pathways that contribute to proliferation (e.g., hypoxia inducible factor 1, nuclear factor kappa B, phosphoinositide 3 kinase/Akt, insulin-like growth factor receptor 1, Wnt, cell cycle associated proteins, as well as androgen and estrogen receptor signaling). These data, in combination with bioinformatics analyses, will be very important for identifying signaling pathways and molecular targets that may provide early diagnostic markers and/or critical targets for the development of new drugs or drug combinations that block tumor formation and progression

Characterization of aminopeptidase encoding gene anp-1 and its association with development in Caenorhabditis elegans

Background Aminopeptidases play important roles in various biological processes in nematodes including growth, development and reproduction. Although the aminopeptidases have been shown to regulate reproduction in Caenorhabditis elegans (C. elegans), the role of aminopeptidases in development and aging has not been reported. This study focused on the function of aminopeptidase AlaNyl aminopeptidase 1 (ANP-1) on development in C. elegans. Methods In the present study, we reported the identification of ANP-1 in C. elegans along with sequence analysis and its functional expression and characterization. The phenotype changes were observed when anp-1 mutated. Then, differential expression genes (DEGs) between wild type strain (N2) and anp-1 deletion strain (RB804) were identified using transcriptome sequencing method. Finally, DEGs were verified by qRT-PCR assay. Results Our observations suggested that anp-1 mutation induced small body size in the L4/young adult stage of C. elegans, however, there was no difference between N2 and RB804 in adult stage. Moreover, deletion of anp-1 resulted in shortening lifespan and laying fewer eggs. DEGs (184 genes) were observed between N2 groups and RB804 groups by transcriptome sequencing. According to GO annotations and KEGG enrichment analysis, these DEGs play vital roles in development regulation in C. elegans. These data demonstrate ANP-1 participates in development and aging of C. elegans and will considerably contribute to the existing knowledge of aminopeptidase function in C. elegans

Neuroligins facilitate the development of bone cancer pain via regulating synaptic transmission: an experimental study

Background: The underlying mechanism of chronic pain involves the plasticity in synaptic receptors and neurotransmitters. This study aimed to investigate potential roles of Neuroligins (NLs) within the spinal dorsal horn of rats in a newly established Bone Cancer Pain (BCP) model. The objective was to explore the mechanism of neuroligin involved in the occurrence and development of bone cancer pain. Methods: Using our rat BCP model, we assessed pain hypersensitivity over time. Quantitative real-time polymerase chain reaction and Western blot analysis were performed to investigate NL expression, and NLs were overexpressed in the rat spinal cord using lentiviral vectors. Immunofluorescence staining and whole-cell patch-clamp recordings were deployed to investigate the role of NLs in the development of BCP. Results: We observed reduced expression levels of NL1 and NL2, but not of NL3, within the rat spinal cord, which were found to be associated with and essential for the development of BCP in our model. Accordingly, NL1 or NL2 overexpression in the spinal cord alleviated mechanical hypersensitivity of rats. Electrophysiological experiments indicated that NL1 and NL2 are involved in BCP via regulating γ-aminobutyric acid-ergic interneuronal synapses and the activity of glutamatergic interneuronal synapses, respectively. Conclusions: Our observations unravel the role of NLs in cancer-related chronic pain and further suggest that inhibitory mechanisms are central features of BCP in the spinal dorsal horn. These results provide a new perspective and basis for subsequent studies elucidating the onset and progression of BCP

Thrombospondin 4, a mediator and candidate indicator of pain

Pain is the most common symptom for which patients seek medical attention. Existing treatments for pain control are largely ineffective due to the lack of an accurate way to objectively measure pain intensity and a poor understanding of the etiology of pain. Thrombospondin 4(TSP4), a member of the thrombospondin gene family, is expressed in neurons and astrocytes and induces pain by interacting with the calcium channel alpha-2-delta-1 subunit (Cavα2δ1). In the present study we show that TSP4 expression level correlates positively with pain intensity, suggesting that TSP4 could be a novel candidate of pain indicator. Using RNAi-lentivirus (RNAi-LV) to knock down TSP4 both in vivo and in vitro, together with electrophysiological experiments involving paired patch-clamp recordings of evoked action potentials and post-synaptic currents in cultured neurons, we found that TSP4 contributes to the development of bone cancer pain, neuropathic pain, and inflammatory pain. This effect is mediated by regulation of neuron excitability via inhibition of synapsin I (Syn I) and modulation of excitatory and inhibitory presynaptic transmission via regulation of vesicular glutamate transporter 2(Vglut2), vesicular GABA transporter (VGAT), and glutamate decarboxylase (GAD) expression. The present study provides a replicable, predictive, valid indicator of pain and demonstrated the underlying molecular and electrophysiological mechanisms by which TSP4 contributes to pain

Darkness increases the population growth rate of the poultry red mite Dermanyssus gallinae

Abstract Background The poultry red mite (PRM), Dermanyssus gallinae, is one of the most economically deleterious ectoparasites affecting egg-laying hens worldwide. It may be possible to control D. gallinae populations by manipulating lighting regimes within poultry units. However, no studies have clearly shown the effects of darkness on the population growth rate of D. gallinae. Methods The effect of darkness on the population growth rate of D. gallinae was investigated, together with the first description of the molecular identity of the mite from China. Mite variables under two lighting regimens (1:23 h L:D and 12:12 h L:D) were compared, including number of mites and eggs, survival and feeding rates, engorgement, oviposition, hatchability and the life-cycle of D. gallinae. Results The results showed that the number of mites (13,763 ± 956) and eggs (5424 ± 317) in the rearing system with prolonged darkness of 1:23 h L:D at 4th week were 2.4- and 3.6-fold higher than those under a conventional lighting regimen of 12:12 h L:D, respectively. The feeding rates of mites under prolonged darkness ranged from 36.7 ± 1.1% to 52.0 ± 7.0%, which were significantly higher than those under conventional lighting regimen (ranging from 22.6 ± 1.9% to 37.3 ± 1.6%). The mean weight of engorged females (0.26 ± 0.01 mg) and the mean number of eggs per female (on average 5.87 ± 0.36) under prolonged darkness were significantly higher than those under conventional lighting regimen (0.22 ± 0.01 mg and 3.62 ± 0.31, respectively). However, the survival rate ranging from 98.07 ± 0.10% to 98.93 ± 0.19%, hatchability of 97.93 ± 0.01% and the life-cycle of D. gallinae (9 days) was not affected by the lighting period. Conclusions Our findings demonstrated that prolonged darkness significantly promoted the proliferation levels of D. gallinae, resulting in increased number of mites and eggs in the rearing system. The promoted population growth of D. gallinae was found to be related to the increased feeding rate, engorgement level and oviposition level of mites under prolonged darkness. The egg hatchability, the survival rates and the duration of life-cycle of D. gallinae were not affected by the light regimes

Bactericidal Efficacy of the Combination of Maresin-like Proresolving Mediators and Carbenicillin Action on Biofilm-Forming Burn Trauma Infection-Related Bacteria

Biofilm-associated bacterial infections are the major reason for treatment failure in many diseases including burn trauma infections. Uncontrolled inflammation induced by bacteria leads to materiality, tissue damage, and chronic diseases. Specialized proresolving mediators (SPMs), including maresin-like lipid mediators (MarLs), are enzymatically biosynthesized from omega-3 essential long-chain polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), by macrophages and other leukocytes. SPMs exhibit strong inflammation-resolving activities, especially inflammation provoked by bacterial infection. In this study, we explored the potential direct inhibitory activities of three MarLs on Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa and Escherichia coli) bacteria in their biofilms that are leading bacteria in burn trauma-related infections. We also examined the effects of MarLs on the bactericidal activities of a typical broad-spectrum antibiotic, carbenicillin (carb), on these bacteria in their preformed biofilms. The results revealed that MarLs combined with carbenicillin can inhibit the survival of Gram-positive and Gram-negative bacteria in their biofilms although MarLs alone did not exhibit bactericidal activity. Thus, our findings suggest that the combination of MarLs and carbenicillin can lower the antibiotic requirements to kill the bacteria in preformed biofilms

Strategies of sRNA identification in <i>Y. pestis</i>.

<p>Strategies of sRNA identification in <i>Y. pestis</i>.</p

Examination of sRNA expression in <i>Y. pestis</i> WT, <i>hfq</i> and <i>crp</i> mutant strain.

<p>(A) Northern blot analysis of sRNA expression in <i>Y. pestis</i> WT (hfq⁺) and <i>hfq</i> mutant strain (hfq⁻). (B) Examination of sRNA expression in <i>Y. pestis</i> WT (crp⁺) and <i>crp</i> mutant strain (crp⁻). (C) Promoter analysis of Yp-sR084. TSS determination by primer extension assay is shown on the left panel. Lanes A, T, C and G represent the Sanger sequencing reactions. The transcription start sites were underlined. Organization and sequence features of sR084 are shown on the right panel. The start sites observed by primer extension are indicated by arrows (+1). Putative -10 and -35 regions of the promoters are boxed. The potential CRP binding sites that match CRP consensus [48] are indicated by dashed lines. The predicted <i>rho</i>-independent terminator is underlined.</p

sRNA detection of <i>Y. pestis</i> grown <i>in vitro</i> or during infection by RNA-seq.

<p>(A) Sequence read distribution of the four cDNA libraries. The read percentage of the cDNA libraries from <i>in </i><i>vitro</i> (TMH and BHI) and <i>in </i><i>vivo</i> (lungs and spleens) samples was indicated, respectively. (B) Sample distribution of sRNAs detected in our study. (C) Comparative analysis of our results with RNA-seq data on <i>Y. pseudotuberculosis</i> from Koo et al. [22] and <i>Y. pestis</i> from Beauregard et al. [23]. The numbers of sRNAs matching with the Rfam database are in brackets.</p

Characterization of sR035 in <i>Y. pestis</i> WT 201 strain.

<p>Transcription start site determination by primer extension assay. Total RNA mixtures from the culture of <i>Y. pestis</i> strain 201 grown in BHI at 26 °C and 37 °C were used to determine TSS by primer extension assays. Lanes T, A, C, and G represent Sanger sequencing reactions. The transcription start sites are underlined. (B) Organization of sequences of sR035. The start sites determined experimentally are indicated by bended arrows 1, 2 and 3. Predicted -10 and -35 regions of the promoters are underlined. The lowercase “a” is the TSS determined by RNA-seq. (C) Distribution of base counts of sR035 among four RNA samples determined by RNA-seq data. The bended arrows 1, 2, 3, and the corresponding vertically broken lines represent the position of TSSs determined by primer extension assays. (D) Measurement of <i>Y. pestis</i> sR035 stability at 26 °C and 37 °C. <i>Y. pestis</i> strain 201 was grown to middle exponential phase in BHI at 26 °C or 37 °C. Rifampicin was added to the cultures and total RNAs were extracted at the time indicated and analyzed by Northern blot analysis (upper panel). The graph shows the relative amount of sR035 remaining at each time point from Northern blotting results as determined by Quantity One software (lower panel). The percentage of the remaining sR035-1 was calculated based on the Northern blot result from total bacterial RNA of <i>Y. pestis</i> grown at 26 °C. The percentages of both sR035-2 and sR035-3 were calculated based on those results at 37 °C.</p