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Chaetostoma chimu sp. nov. is described from 119 specimens collected in the Andean foothills of the Orinoco River in Colombia. The new species can be distinguished from congeners by a combination of characters, including shape of cheek odontodes, presence of a supraoccipital excrescence, sexual dimorphism characters and colour pattern. In addition, this species can be distinguished from the sympatric and geographically close congeners of the Orinoco that have curved cheek odontodes (i.e., Chaetostoma anale, Chaetostoma formosae and Chaetostoma joropo), by presenting a generally larger orbital diameter and interorbital distance, details of sexual dimorphism and colour pattern. The type locality of specimens collected by Kjell von Sneidern in the mid-twentieth century in the Colombian Amazon is also clarified. Chaetostoma alternifasciatum and Chaetostoma vagum are proposed as junior synonyms of C. anale. An identification key for the species of Chaetostoma from cis-Andean drainages in Colombia is provided. The new species is herein categorized as Vulnerable (Vu) B1a (i, iii), following the IUCN criteria.

Moenkhausia hemigrammoides Géry, 1965 was described from Suriname and recently found in the Ariari drainage, part of the Orinoco river basin. Here, we formally report the presence of this species in the Ariari and Inírida drainages of the Guaviare River and present morphological data that complement the original description. Our records of M. hemigrammoides represent the westernmost distribution for this Guianan species in the Amazonas–Orinoco transition zone.

A new species of Spatuloricaria is described from the Orinoco River basin, Colombia. The new species can be distinguished from its congeners by the following characters: a broad, dark brown stripe on the first pre-dorsal plate, which occupies the entire plate, reaching the preopercle, and sometimes reaching the second pre-dorsal plate; the possession of a small group of plates posterior to the urogenital pore; the abdominal surface with scattered, very small plates leaving naked areas; the possession of five transverse dark brown bands on the dorsal region; and four to five premaxillary teeth. The new species is the first valid species of Spatuloricaria distributed in the Orinoco River basin. Ecological notes of the species and comments regarding the taxonomy and phylogenetic relationships of the genus are offered.

Chrysobrycon mojicai sp. nov. is described from forest streams that are part of the Amazon River basin in Colombia. It is distinguished from all congeners by having most of the maxillary teeth with distal tips lateroventrally curved (v. teeth straight along their lengths) and a greater number of dentary teeth (20-27 v. 11-19, except in Chrysobrycon yoliae). The posterior margin of the ventral process of the quadrate does not reach the vertical through the posterior margin of the symplectic and these features differentiate C. mojicai from Chrysobrycon myersi and C. yoliae (v. posterior margin of ventral process of quadrate reaching vertical through posterior margin of symplectic). In species of Chrysobrycon, the frontals are extensively contacting each other along the midline, resulting in an absent frontal fontanel and a reduced parietal fontanel. This extensive contact between the frontals modifying the fontanels is a condition rarely found within the Stevardiinae and hence is proposed as an additional diagnostic characteristic for the genus. An updated identification key for all Chrysobrycon species is provided.

Hyphessobrycon klausanni sp. n. is described from small drainages of the upper Guaviare River (Orinoco River Basin) in Colombia. It differs from all congeners by having a wide, conspicuous, dark lateral stripe extending from the anterior margin of the eye across the body and continued through the middle caudal-fin rays, and that covers (vertically) three or four horizontal scale rows. It also differs by having an orange-yellow stripe extending from the anterosuperior margin of the eye to the caudal peduncle above the lateral line in life. It differs from all other species of Hyphessobrycon that have a similar dark lateral stripe: H. cyanotaenia, H. loretoensis, H. melanostichos, H. nigricinctus, H. herbertaxelrodi, H. eschwartzae, H. montogoi, H. psittacus, H. metae, H. margitae, H. vanzolinii, and H. peruvianus in having only three or four pored scales in the lateral line, 21 to 24 lateral scales and six teeth in the inner premaxillary row. Hyphessobrycon klausanni differs from H. loretoensis in having seven to eight maxillary teeth (vs. three to four) and in having a longer caudal peduncle (12.4–17.0% SL vs. 4.6–8.0% SL). Additionally Hyphessobrycon klausanni can be distinguished from the other species of Hyphessobrycon with a dark lateral stripe from the Orinoco River Basin (H. metae and H. acaciae) in having two teeth in the outer premaxillary row (vs. three to four) and 10 branched pectoral–fin rays (vs. 11 to 12). It further differs from H. metae by the length of the snout (17.6–22.8% HL vs. 9.9–15.2% HL) and by the length of the caudal peduncle (12.4–17.0% SL vs. 7.3–11.8% SL).

The present study describes ecological aspects of Lebiasina erythrinoides; 200 individuals were sampled throughout an annual hydrological cycle from October 2008 to September 2009 in La Calaboza stream, a Piedmont tributary of the Rio Cravo Sur. The studied population had more females than males (1.5:1) and preferred habitats dominated by shoreline vegetation and rocky substrates. Prior to reproductive events, individuals presented an increase in the condition factor (K) and the gonosomatic index (GSI). On average, 648.8 oocytes are discharged twice a year: at the beginning of the rainy season, and during falling water phase. The mean sizes at sexual maturity found for females were 73.5 mm standard length (SL) and for males 70 mm SL. Rounding up, an average minimum size of capture of 75 mm SL is proposed. Most specimens of the population (65 %) were found to be maturing or mature (stages II and III), 28.5 % immature (stage I), 5.0 % in post reproduction stage V, and 1.5 % in reproductive phase (IV). Results suggest this species is omnivorous with a preference for invertebrates (IRI = 41.2 %) and vegetal material (IRI = 27.8 %), but also includes a variety of other items. The values of the diet are correlated with hydrological cycle and size.

Hyphessobrycon klausanni sp. n. is described from small drainages of the upper Guaviare River (Orinoco River Basin) in Colombia. It differs from all congeners by having a wide, conspicuous, dark lateral stripe extending from the anterior margin of the eye across the body and continued through the middle caudal-fin rays, and that covers (vertically) three or four horizontal scale rows. It also differs by having an orange-yellow stripe extending from the anterosuperior margin of the eye to the caudal peduncle above the lateral line in life. It differs from all other species of Hyphessobrycon that have a similar dark lateral stripe: H. cyanotaenia, H. loretoensis, H. melanostichos, H. nigricinctus, H. herbertaxelrodi, H. eschwartzae, H. montogoi, H. psittacus, H. metae, H. margitae, H. vanzolinii, and H. peruvianus in having only three or four pored scales in the lateral line, 21 to 24 lateral scales and six teeth in the inner premaxillary row. Hyphessobrycon klausanni differs from H. loretoensis in having seven to eight maxillary teeth (vs. three to four) and in having a longer caudal peduncle (12.4–17.0% SL vs. 4.6–8.0% SL). Additionally Hyphessobrycon klausanni can be distinguished from the other species of Hyphessobrycon with a dark lateral stripe from the Orinoco River Basin (H. metae and H. acaciae) in having two teeth in the outer premaxillary row (vs. three to four) and 10 branched pectoral–fin rays (vs. 11 to 12). It further differs from H. metae by the length of the snout (17.6–22.8% HL vs. 9.9–15.2% HL) and by the length of the caudal peduncle (12.4–17.0% SL vs. 7.3–11.8% SL).

Chaetostoma joropo n. sp. is described from the piedmont of the Orinoco River drainage in Colombia. The new species has been long confused with Chaetostoma milesi, a species with similar overall morphology and color pattern that is restricted to the Magdalena-Cauca River Basin. We diagnose the new species on the basis of morphology as well as a precise description of the color pattern. Chaetostoma joropo n. sp. is also easily distinguished from C. formosae the most similar species and other species inhabiting the Orinoco River drainage in Colombia. Data on ontogenetic variation and sexual dimorphism are provided, as well as natural history notes and remarks on the usage of the name Chaetostoma milesi for specimens from both the Orinoco and Magdalena-Cauca drainages. A discussion on the usage of the name Chaetostoma platyrhynchus is also provided given its current instability in the literature.

Apoptosis-inducing factor (AIF) exhibits reactive oxygen species (ROS)-generating NADH oxidase activity of unknown significance, which is dispensable for apoptosis. We knocked out the aif gene in two human colon carcinoma cell lines that displayed lower mitochondrial complex I oxidoreductase activity and produced less ROS, but showed increased sensitivity to peroxide- or drug-induced apoptosis. AIF knockout cells failed to form tumors in athymic mice or grow in soft agar. Only AIF with intact NADH oxidase activity restored complex I activity and anchorage-independent growth of aif knockout cells, and induced aif-transfected mouse NIH3T3 cells to form foci. AIF knockdown in different carcinoma cell types resulted in lower superoxide levels, enhanced apoptosis sensitivity and loss of tumorigenicity. Antioxidants sensitized AIF-expressing cells to apoptosis, but had no effect on tumorigenicity. In summary, AIF-mediated resistance to chemical stress involves ROS and probably also mitochondrial complex I. AIF maintains the transformed state of colon cancer cells through its NADH oxidase activity, by mechanisms that involve complex I function. On both counts, AIF represents a novel type of cancer drug target.

The high-affinity human interleukin-7 (IL-7)R is a heterodimeric complex consisting of the IL-7Ralpha and common interleukin-2 receptor gamma (IL-2Rgamma(c)) chains. Activation of the IL-7R complex is associated with tyrosine and serine residue phosphorylation of a number of intracellular substrates leading to proliferation and induction of various cellular differentiation processes. In this study, we demonstrate, by S1 nuclease protection assay, immunoprecipitation and in vitro kinase assay that functional human (h) IL-7R is expressed in haematopoietic and nonhaematopoietic cell lines. The National Cancer Institute (NCI) tumour panel of 60 cell lines (NCI60) was screened for the expression of IL-7R mRNA by S1 nuclease protection assay, and IL-7R mRNA was detected in 9 of 12 leukemia, 3 of 7 lung, 4 of 6 CNS, 2 of 7 melanoma, 2 of 7 renal, 1 of 6 colon and 1 of 6 breast cancer cell lines. Immunoblot analysis of haematopoietic, lung cancer and brain tumour cell lines demonstrated expression of IL-7R, IL-2Rgamma(c) and p59 fyn, suggesting that the components of an IL-7R signalling network are present in nonhaematopoietic neoplastic cells. Immunoprecipitation of IL-7Ralpha followed by an in vitro kinase assay demonstrated functional receptor phosphorylation events in the lung cancer cells but not in the brain tumour cell lines. The expression of functional IL-7R on epithelial tumour cells may represent a potential target for receptor-directed therapy.

AG957 (NSC 654705) is a tyrphostin tyrosine kinase inhibitor that has been demonstrated previously to induce growth arrest in chronic myelogenous leukemia cells by inhibiting p210 bcr–abl kinase activity and by stabilizing the association of p210 bcr–abl kinase with its signaling adaptor molecules, Shc and Grb2. In previous studies, it has been demonstrated that AG957-associated down-regulation of bcr–abl activates the cytochrome c /Apaf-1/caspase-9 pathway and induces apoptosis in chronic myelogenous leukemia blasts and progenitor cells. While AG957 has been purported to have specificity for the p210 bcr–abl kinase, antiproliferative effects of AG957 in normal T-lymphocytes and bcr–abl negative leukemia cells suggest that other targets, such as c-CBL, may be substrates. In this study, we explored the mechanisms of AG957-mediated growth inhibition and apoptosis in the p210 bcr–abl negative leukemia cell lines Nalm-6 and Jurkat, and demonstrate that AG957-mediated apoptosis is associated with altered phosphorylation of Akt and BAD, which destabilizes the Bcl-xL/BAD complex and releases the block to apoptosis. We, therefore, propose that AG957 induces apoptosis in bcr–abl negative hematopoietic cells by affecting the phosphorylation state of phosphatidylinositol-3 kinase/Akt.

Chaetostoma joropo n. sp. is described from the piedmont of the Orinoco River drainage in Colombia. The new species has been long confused with Chaetostoma milesi, a species with similar overall morphology and color pattern that is restricted to the Magdalena-Cauca River Basin. We diagnose the new species on the basis of morphology as well as a precise description of the color pattern. Chaetostoma joropo n. sp. is also easily distinguished from C. formosae the most similar species and other species inhabiting the Orinoco River drainage in Colombia. Data on ontogenetic variation and sexual dimorphism are provided, as well as natural history notes and remarks on the usage of the name Chaetostoma milesi for specimens from both the Orinoco and Magdalena-Cauca drainages. A discussion on the usage of the name Chaetostoma platyrhynchus is also provided given its current instability in the literature.

HAMLET is the first member of a new family of tumoricidal protein-lipid complexes that kill cancer cells broadly, while sparing healthy, differentiated cells. Many and diverse tumor cell types are sensitive to the lethal effect, suggesting that HAMLET identifies and activates conserved death pathways in cancer cells. Here we investigated the molecular basis for the difference in sensitivity between cancer cells and healthy cells. Using a combination of small hairpin RNA inhibition, proteomic and metabolomic technology we identified the c-Myc oncogene as one essential determinant of HAMLET sensitivity. Increased c-Myc expression levels promoted the sensitivity to HAMLET and shRNA knockdown of c-Myc suppressed the lethal response, suggesting that oncogenic transformation with c-Myc creates a HAMLET-sensitive phenotype. Furthermore, the HAMLET sensitivity was modified by the glycolytic state of the tumor cells. Glucose deprivation sensitized tumor cells to HAMLET-induced cell death and in the shRNA screen Hexokinase 1, PFKFB1 and HIF1α modified HAMLET sensitivity. Hexokinase 1 was shown to bind HAMLET in a protein array containing approximately 8000 targets and Hexokinase activity decreased within 15 minutes of HAMLET treatment, prior to morphological signs of tumor cell death. In parallel, HAMLET triggered rapid metabolic paralysis in carcinoma cells. The glycolytic machinery was modified and glycolysis was shifted towards the pentose phosphate pathway. Tumor cells were also shown to contain large amounts of oleic acid and its derivatives already after 15 minutes. The results identify HAMLET as a novel anti-cancer agent that kills tumor cells by exploiting unifying features of cancer cells such as oncogene-addiction or the Warburg effect.

Cell adhesion is tightly regulated by specific molecular interactions and detachment from the extracellular matrix modifies proliferation and survival. HAMLET (Human Alpha-lactalbumin Made LEthal to Tumor cells) is a protein-lipid complex with tumoricidal activity that also triggers tumor cell detachment in vitro and in vivo, suggesting that molecular interactions defining detachment are perturbed in cancer cells. To identify such interactions, cell membrane extracts were used in Far-western blots and HAMLET was shown to bind α-actinins; major F-actin cross-linking proteins and focal adhesion constituents. Synthetic peptide mapping revealed that HAMLET binds to the N-terminal actin-binding domain as well as the integrin-binding domain of α-actinin-4. By co-immunoprecipitation of extracts from HAMLET-treated cancer cells, an interaction with α-actinin-1 and -4 was observed. Inhibition of α-actinin-1 and α-actinin-4 expression by siRNA transfection increased detachment, while α-actinin-4-GFP over-expression significantly delayed rounding up and detachment of tumor cells in response to HAMLET. In response to HAMLET, adherent tumor cells rounded up and detached, suggesting a loss of the actin cytoskeletal organization. These changes were accompanied by a reduction in β1 integrin staining and a decrease in FAK and ERK1/2 phosphorylation, consistent with a disruption of integrin-dependent cell adhesion signaling. Detachment per se did not increase cell death during the 22 hour experimental period, regardless of α-actinin-4 and α-actinin-1 expression levels but adherent cells with low α-actinin levels showed increased death in response to HAMLET. The results suggest that the interaction between HAMLET and α-actinins promotes tumor cell detachment. As α-actinins also associate with signaling molecules, cytoplasmic domains of transmembrane receptors and ion channels, additional α-actinin-dependent mechanisms are discussed.

The mucosal immune system identifies and fights invading pathogens, while allowing non-pathogenic organisms to persist. Mechanisms of pathogen/non-pathogen discrimination are poorly understood, as is the contribution of human genetic variation in disease susceptibility. We describe here a new, IRF3-dependent signaling pathway that is critical for distinguishing pathogens from normal flora at the mucosal barrier. Following uropathogenic E. coli infection, Irf3−/− mice showed a pathogen-specific increase in acute mortality, bacterial burden, abscess formation and renal damage compared to wild type mice. TLR4 signaling was initiated after ceramide release from glycosphingolipid receptors, through TRAM, CREB, Fos and Jun phosphorylation and p38 MAPK-dependent mechanisms, resulting in nuclear translocation of IRF3 and activation of IRF3/IFNβ-dependent antibacterial effector mechanisms. This TLR4/IRF3 pathway of pathogen discrimination was activated by ceramide and by P-fimbriated E. coli, which use ceramide-anchored glycosphingolipid receptors. Relevance of this pathway for human disease was supported by polymorphic IRF3 promoter sequences, differing between children with severe, symptomatic kidney infection and children who were asymptomatic bacterial carriers. IRF3 promoter activity was reduced by the disease-associated genotype, consistent with the pathology in Irf3−/− mice. Host susceptibility to common infections like UTI may thus be strongly influenced by single gene modifications affecting the innate immune response., Author Summary The host immune system must identify pathogens and defeat them through TLR-dependent signaling pathway activation, while distinguishing them from commensal flora. Contrary to current dogma, the host cannot solely use “pattern recognition” since the microbial molecules involved in such recognition are present on pathogens and commensals alike. We identify here a pathogen-specific mechanism of TLR4 activation and signaling intermediates in this pathway, leading to IRF3-dependent transcription of innate immune response genes. We show in knockout mice that Irf3 deficiency causes severe tissue pathology and that effector functions controlled by IFNβ are involved. Finally, in highly disease-prone pyelonephritis patients we found a high frequency of IRF3 promoter polymorphism compared to asymptomatic bacterial carriers or controls. The polymorphisms influenced promoter activity in reporter assays, suggesting that they are functionally important. Urinary tract infections are among the most common bacterial infections in man, and are a major cause of morbidity and mortality. A subset of disease-prone individuals is at risk for recurrent disease, severe renal dysfunction and end-stage renal disease. At present, there is no method to identify disease-prone infants and to prevent future morbidity and renal damage. The genetic and functional studies described here indicate that genetic variation in IRF3 influences individual susceptibility to kidney infection and might serve as a new tool for future risk assessment in this patient group.

Apoptosis-inducing factor (AIF) is expelled from mitochondria after some apoptotic stimuli and translocates to the nucleus, which may contribute to DNA and nuclear fragmentation in some non-physiological mammalian cell deaths. Conversely, the requirement for mitochondrial AIF in oxidative phosphorylation and energy generation provides a plausible explanation for the embryonic lethality or neurodegeneration that has been found in different AIF-deficient mouse models. These findings may help illuminate the ability of mitochondrial AIF to suppress cytoplasmic stress granule formation and to promote the tumorigenic growth of cancer cells. AIF is ideally located in the mitochondrion to perform a vital normal function in energy production. Once it translocates to the nucleus, however, the cell might die either of energy failure or nuclear fragmentation (or both). We propose that the main function of AIF is to support energy production in both normal and transformed cell physiology, whereas nuclear-translocated AIF might contribute to stress-induced or pathological cell death in certain scenarios.

DAB(389)IL-2 (ONTAK) is a fusion protein consisting of the ADP-ribosyltransferase and membrane translocating domains of native diphtheria toxin and the full-length sequence for interleukin-2 (IL-2) gene. In vitro data demonstrates that DAB(389)IL-2 is cytotoxic to cells expressing the high affinity IL-2 receptor (IL-2R). In Phases I and II clinical trials of patients whose tumor cells express a component of the IL-2R, the response rates were 18% for B-cell non-Hodgkin lymphoma (NHL) and 30% for cutaneous T-cell lymphoma (CTCL). In this study, we examined the effects of arginine butyrate on IL-2R expression and susceptibility of leukemia cells to intoxication by DAB(389)IL-2. We demonstrate that the p75 subunit of the IL-2R (IL-2Rbeta) is upregulated in the presence of low concentrations of arginine butyrate (0.06mM) which had no direct growth inhibitory effect on the cells. To explore mechanisms of this upregulation, we examined the effect of 0.06mM arginine butyrate on relevant transcriptional elements and on histone deacetylase and found activation of cAMP response element (CRE) but not NFAT or NFKB, as well as inhibition of histone deacetylase (HDAC). Our results suggest that the effects of physiologically achievable concentrations of butyrate on IL-2R expression could be exploited to enhance the susceptibility of intermediate and low-affinity IL-2R expressing leukemia cells to DAB(389)IL-2.

Endonucleolytic DNA fragmentation is the common end point and the prevailing indicator of apoptosis. We have identified a 70-kDa endonuclease (NUC70) that is activated in drug-induced apoptosis of human hematopoietic cells. We purified NUC70 to homogeneity and generated a rabbit polyclonal antibody to distinguish it from previously identified nucleases. Biochemical characterization of isolated NUC70 demonstrates that it is Ca2+/Mg2+-dependent and active over a pH range of 6-8. When incubated with isolated HeLa nuclei, NUC70 was capable of generating internucleosomal DNA fragmentation. This endonucleolytic activity was inhibited by Zn2+, aurintricarboxylic acid, N-ethylmaleimide, spermine, and iodoacetamide. Western immunoblots using the anti-NUC70 antibody and DNA-SDS-polyacrylamide gel electrophoresis assays indicate that NUC70 expression and activity is restricted to human hematopoietic cells. No such activity was detected in human epithelial cell lines or murine hematopoietic cells. We also observed no difference in levels of NUC70 expression between apoptotic and nonapoptotic cells, suggesting that activation of NUC70 may be by posttranslational modification. We demonstrate that NUC70 activity is diminished in cells pretreated with the caspase inhibitors z-DEVD-fmk, z-VAD-fmk, and Z-CH2-Asp-DCB. Time course studies of cytoplasmic and nuclear endonuclease activities during apoptosis show that NUC70 is a cytoplasmic endonuclease that is translocated to the nucleus after the initiation of apoptosis. We confirmed this with immunostaining studies using anti-NUC70 antibody. These results demonstrate that NUC70 is an endogenous cytoplasmic endonuclease that is activated during apoptosis in a caspase-dependent mechanism.

Currently, only 20 fish species have been recorded from the Casanare River, which is one of the main tributaries of the Meta River drainage. To characterize the fish assemblages of this river, samplings were carried out at 35 collecting stations in low and high water seasons of 2015. The results obtained, raised the number of fish species of the Casanare River drainage to 180, of which 41 are exclusive to the piedmont, 83 to the savannah and 56 are shared between both units; 19 species are endemic, 11 are migratory, and three are classified in one of the national threat categories. Three species were found to be undescribed and one represents a new record for the Orinoco River Basin in Colombia. This study fills the lack of information for the Casanare River drainage, highlights its importance as a potential area for biodiversity conservation and facilitates the decision-making process related to the management planning and conservation of the hydrographic basin.