Suppression of oxidative phosphorylation confers resistance against bevacizumab in experimental glioma

Although bevacizumab initially shows high response rates in gliomas and other tumours, therapy resistance usually develops later. Because anti‐angiogenic agents are supposed to induce hypoxia, we asked whether rendering glioma cells independent of oxidative phosphorylation modulates their sensitivity against hypoxia and bevacizumab. LNT‐229 glioma cells without functional mitochondria (rho⁰) and control (rho⁺) cells were generated. LNT‐229 rho⁰‐cells displayed reduced expression of oxidative phosphorylation‐related genes and diminished oxygen consumption. Conversely, glycolysis was up‐regulated in these cells, as shown by increased lactate production and stronger expression of glucose transporter‐1 and lactate dehydrogenase‐A. However, hypoxia‐induced cell death in vitro was nearly completely abolished in the LNT‐229 rho⁰‐cells, these cells were more sensitive towards glucose restriction and the treatment with the glycolysis inhibitor 2‐deoxy‐D‐glucose. In an orthotopic mouse xenograft experiment, bevacizumab induced hypoxia as reflected by elevated Hypoxia‐inducible factor 1‐alpha staining in both, rho⁺‐ and rho⁰‐tumours. However, it prolonged survival only in the mice bearing rho⁺‐tumours (74 days vs. 105 days, p  = 0.024 log‐rank test) and had no effect on survival in mice carrying LNT‐229 rho⁰‐tumours (75 days vs. 70 days, p  = 0.52 log‐rank test). Interestingly, inhibition of glycolysis in vivo with 2‐deoxy‐D‐glucose re‐established sensitivity of rho⁰‐tumours against bevacizumab (98 days vs. 80 days, p  = 0.0001). In summary, ablation of oxidative phosphorylation in glioma cells leads to a more glycolytic and hypoxia‐resistant phenotype and is sufficient to induce bevacizumab‐refractory tumours. These results add to increasing evidence that a switch towards glycolysis is one mechanism how tumour cells may evade anti‐angiogenic treatments and suggest anti‐glycolytic strategies as promising approaches to overcome bevacizumab resistance.

     

Effects of parental care on the accumulation and release of cryptic genetic variation: <Emphasis Type="Italic">review of mechanisms and a case study of dung beetles</Emphasis>

Cryptic genetic variation plays an important role in the emergence of disease and evolutionary responses to environmental change. Focusing on parental care behavior, we discuss three mechanisms by which behavior can affect the accumulation and release of cryptic genetic variation. We illustrate how these hypotheses might be tested with preliminary data from Onthophagus dung beetles, which provide indirect parental care by provisioning their offspring with dung and sheltering them underground. The environmental stress hypothesis states that parental care reduces selection intensity on novel mutations when increased parental care results in a less stressful offspring environment. A review of recent literature, coupled with an irradiation experiment in beetles, suggests this mechanism may operate in some situations, but depends on the types of mutations under consideration. The relaxed selection hypothesis states that genes expressed in low care environments should be under weakened selection because their phenotypic manifestations are exposed to selection less frequently, and thus are prone to mutation accumulation. If parental care is reduced, for instance due to population-wide environmental changes, such cryptic variation may exert phenotypic effects, becoming exposed to selection. There is substantial theory in support of this hypothesis, and comparisons between beetle populations that differ in parental care behavior further support this idea. Finally, the compensation hypothesis states that organisms with direct parental care may be able to respond to cues or signals from offspring and compensate for genetic variants. We highlight the extensive discussion of this hypothesis with respect to medical care and genetic load in humans and explore invertebrate systems that may constitute powerful models for further inquiry. In summary, several mechanisms exist by which care behavior may shape the accumulation and release of cryptic genetic variation, thereby affecting the potential emergence of diseases and the rate and direction of evolutionary responses to novel environments.     

A premature termination codon in either exon of minute virus of mice P4 promoter-generated pre-mRNA can inhibit nuclear splicing of the intervening intron in an open reading frame-dependent manner.

How premature translation termination codons (PTCs) mediate effects on nuclear RNA processing is unclear. Here we show that a PTC at nucleotide (nt) 385 in the NS1/2 shared exon of P4-generated pre-mRNAs of the autonomous parvovirus minute virus of mice caused a decrease in the accumulated levels of doubly spliced R2 relative to singly spliced R1, although the total accumulated levels of R1 plus R2 remained the same. The effect of this PTC was evident within nuclear RNA, was mediated by a PTC and not a missense transversion mutation at this position, and could be suppressed by improvement of the large intron splice sites and by mutation of the AUG that initiated translation of R1 and R2. In contrast to the PTC at nt 385, the reading frame-dependent effect of the PTC at nt 2018 depended neither on the initiating AUG nor the normal termination codon for NS2; however, it could be suppressed by a single nucleotide deletion mutation in the upstream NS1/2 common exon that shifted the 2018 PTC out of the NS2 open reading frame. This suggested that there was recognition and communication of reading frame between exons on a pre-mRNA in the nucleus prior to or concomitant with splicing.     

Mechanical stimulation of osteopontin mRNA expression and synthesis in bone cell cultures

We have shown earlier that mechanical stimulation by intermittent hydrostatic compression (IHC) promotes alkaline phosphatase and procollagen type I gene expression in calvarial bone cells. The bone matrix glycoprotein osteopontin (OPN) is considered to be important in bone matrix metabolism and cell-matrix interactions, but its role is unknown. Here we examined the effects of IHC (13 kPa) on OPN mRNA expression and synthesis in primary calvarial cell cultures and the osteoblast-like cell line MC3T3-E1. OPN mRNA expression declined during control culture of primary calvarial cells, but not MC3T3-E1 cells. IHC upregulated OPN mRNA expression in late released osteoblastic cell cultures, but not in early released osteoprogenitor-like cells. Also, in both proliferating and differentiating MC3T3-E1 cells, OPN mRNA expression and synthesis were enhanced by IHC, differentiating cells being more responsive than proliferating cells. These results suggest a role for OPN in the reaction of bone cells to mechanical stimuli. The severe loss of OPN expression in primary bone cells cultured without mechanical stimulation suggests that disuse conditions down-regulate the differentiated osteoblastic phenotype. J. Cell. Physiol. 170:174–181, 1997. © 1997 Wiley-Liss, Inc.     

THE GENETIC BASIS OF ZEBRA FINCH VOCALIZATIONS

Animal vocalizations play an important role in individual recognition, kin recognition, species recognition, and sexual selection. Despite much work in these fields done on birds virtually nothing is known about the heritability of vocal traits in birds. Here, we study a captive population of more than 800 zebra finches ( Taeniopygia guttata ) with regard to the quantitative genetics of call and song characteristics. We find very high heritabilities in nonlearned female call traits and considerably lower heritabilities in male call and song traits, which are learned from a tutor and hence show much greater environmental variance than innate vocalizations. In both sexes, we found significant heritabilities in several traits such as mean frequency and measures of timbre, which reflect morphological characteristics of the vocal tract. These traits also showed significant genetic correlations with body size, as well as positive genetic correlations between the sexes, supporting a scenario of honest signaling of body size through genetic pleiotropy ("index signal"). In contrast to such morphology-related voice characteristics, classical song features such as repertoire size or song length showed very low heritabilities. Hence, these traits that are often suspected to be sexually selected would hardly respond to current directional selection.     

Risk Assessment for Dermal Exposure of Organochlorine Pesticides for Local Fishermen in the Rangsit Agricultural Area,Central Thailand

Organochlorine pesticides (OCPs) used in agriculture and for public health purposes were banned in Thailand over the past decade; however, their persistent residues have been found in several agricultural areas of the country. This may result in adverse effects to human populations. This study investigated the concentration of organochlorine pesticides residues (OCPRs) in surface water and evaluated the potential cancer risk associated with dermal contact of the local fisherman fishing in the Khlong 7 canal, Rangsit agricultural area, central Thailand. Water samples were extracted using liquid-liquid extraction (LLE) and then analyzed by gas chromatography with microelectron capture detector (μ -ECD). The results show that low concentrations of OCPRs were detected in parts per billion (ppb or ng/ml) levels, that is, ∑ Endosulfan (α -, β -, and -sulfate) 0.082 ng/ml > DDT and derivatives 0.019 ng/ml > ∑ HCH (α -, γ -, β -, and δ -HCH) 0.014 ng/ml > aldrin and dieldrin 0.007 ng/ml > heptachlor and heptachlor epoxide 0.0068 ng/ml > endrin and endrin aldehyde 0.005 ng/ml > methoxychlor 0.001 ng/ml, respectively. Using the worst-case scenario defined as the reasonable maximum exposure (RME) to assess the potential cancer risk, five OCPs (dieldrin, 4,4′ -DDT, β -HCH, heptachlor, and heptachlor epoxide) may pose a risk of concern on a lifetime human carcinogenesis greater than one in a million.