No correlative evidence of costs of infection or immunity on leucocyte telomere length in a wild population of Soay sheep

Telomere length (TL) is a biomarker hypothesized to capture evolutionarily and ecologically important physiological costs of reproduction, infection and immunity. Few studies have estimated the relationships among infection status, immunity, TL and fitness in natural systems. The hypothesis that short telomeres predict reduced survival because they reflect costly consequences of infection and immune investment remains largely untested. Using longitudinal data from a free-living Soay sheep population, we tested whether leucocyte TL was predicted by infection with nematode parasites and antibody levels against those parasites. Helminth parasite burdens were positively associated with leucocyte TL in both lambs and adults, which is not consistent with TL reflecting infection costs. We found no association between TL and helminth-specific IgG levels in either young or old individuals which suggests TL does not reflect costs of an activated immune response or immunosenescence. Furthermore, we found no support for TL acting as a mediator of trade-offs between infection, immunity and subsequent survival in the wild. Our results suggest that while variation in TL could reflect short-term variation in resource investment or environmental conditions, it does not capture costs of infection and immunity, nor does it behave like a marker of an individual's helminth-specific antibody immune response

Radiation-induced biologic bystander effect elicited in vitro by targeted radiopharmaceuticals labeled with alpha-, beta-, and Auger electron-emitting radionuclides

Recent studies have shown that indirect effects of ionizing radiation may contribute significantly to the effectiveness of radiotherapy by sterilizing malignant cells that are not directly hit by the radiation. However, there have been few investigations of the importance of indirect effects in targeted radionuclide treatment. Our purpose was to compare the induction of bystander effects by external beam gamma-radiation with those resultant from exposure to 3 radiohaloanalogs of metaiodobenzylguanidine (MIBG): I-131-MIBG (low-linear-energy-transfer [LET] beta-emitter), I-123-MIBG (potentially high-LET Auger electron emitter), and meta-At-211-astatobenzylguanidine (At-211-MABG) (high-LET a-emitter). Methods: Two human tumor cell lines-UVW (glioma) and EJ138 (transitional cell carcinoma of bladder)-were transfected with the noradrenaline transporter (NAT) gene to enable active uptake of MIBG. Medium from cells that accumulated the radiopharmaceuticals or were treated with external beam radiation was transferred to cells that had not been exposed to radioactivity. and clonogenic survival was determined in donor and recipient cultures. Results: Over the dose range 0-9 Gy of external beam radiation of donor cells, 2 Gy caused 30%-40% clonogenic cell kill in recipient cultures. This potency was maintained but not increased by higher dosage. In contrast, no corresponding saturation of bystander cell kill was observed after treatment with a range of activity concentrations of 131 I-MIBG, which resulted in up to 97% death of donor cells. Cellular uptake of (123I)-MIBG and At-211-MABG induced increasing recipient cell kill up to levels that resulted in direct kill of 35%-70% of clonogens. Thereafter, the administration of higher activity concentrations of these high-LET emitters was inversely related to the kill of recipient cells. Over the range of activity concentrations examined, neither direct nor indirect kill was observed in cultures of cells not expressing the NAT and, thus, incapable of active uptake of MIBG. Conclusion: Potent toxins are generated specifically by cells that concentrate radiohalogenated MIBG. These may be LET dependent and distinct from those elicited by conventional radiotherapy

An efficient targeted radiotherapy/gene therapy strategy utilising human telomerase promoters and radioastatine and harnessing radiation-mediated bystander effects

Background<br/>Targeted radiotherapy achieves malignant cell-specific concentration of radiation dosage by tumour-affinic molecules conjugated to radioactive atoms. Combining gene therapy with targeted radiotherapy is attractive because the associated cross-fire irradiation of the latter induces biological bystander effects upon neighbouring cells overcoming low gene transfer efficiency.<br/> Methods<br/>We sought to maximise the tumour specificity and efficacy of noradrenaline transporter (NAT) gene transfer combined with treatment using the radiopharmaceutical meta-[<sup>131</sup>1] iodobenzylguanidine ([<sup>131</sup>1]MIBG). Cell-kill was achieved by treatment with the beta-decay particle emitter [<sup>131</sup>1]MIBG or the alpha-particle emitter [<sup>211</sup>At]MABG. We utilised our novel transfected mosaic spheroid model (TMS) to determine whether this treatment strategy could result in sterilisation of spheroids containing only a small proportion of NAT-expressing cells.<br/> Results<br/>The concentrations of [<sup>131</sup>1]MIBG and [<sup>211</sup>At]MABG required to reduce to 0.1% the survival of clonogens derived from the TMS composed of 100% of NAT gene-transfected cells were 1.5 and 0.004 MBq/ml (RSV promoter), 8.5 and 0.0075 MBq/ml (hTR promoter), and 9.0 and 0.008 MBq/ml (hTERT promoter), respectively. The concentrations of radiopharmaceutical required to reduce to 0.1% the survival of clonogens derived from 5% RSV/NAT and 5% hTERT/NAT TMS were 14 and 23 MBq/ml, respectively, for treatment with [<sup>131</sup>1]MIBG and 0.018 and 0.028 MBq/ml, respectively, for treatment with [<sup>211</sup>At]MABG.<br/> Conclusions<br/>These results indicate that the telomerase promoters have the capacity to drive the expression of the NAT. The potency of [<sup>211</sup>At]MABG is approximately three orders of magnitude greater than that of [<sup>131</sup>1]MIBG. Spheroids composed of only 5% of cells expressing NAT under the control of the RSV or hTERT promoter were sterilised by radiopharmaceutical treatment. This observation is indicative of bystander cell-kill

Efeito da oferta dietética de proteína sobre o ganho muscular, balanço nitrogenado e cinética da 15N-glicina de atletas em treinamento de musculação Effect of the dietary protein intake on the muscular gain, nitrogen balance and 15N-glycine kinetics of athletes in resistance training

O efeito da oferta crescente de proteína sobre o ganho muscular, balanço nitrogenado e cinética da 15N-glicina de atletas de musculação foi estudado em seis jovens saudáveis, praticantes de treinamento com pesos (> 2 anos), sem uso de anabolizantes e concordes com os princípios éticos da pesquisa. Todos receberam adequações dietéticas (0,88g de proteína/kg/dia) pré-experimento de 2 semanas (D1) após o que se ofereceu, por idêntico período, dieta contendo 1,5g de proteína/kg de peso corporal/dia com 30kcal/g de proteína (dieta D2). A seguir receberam, nas próximas 2 semanas, a dieta D3, contendo 2,5g de proteína/kg de peso corporal/dia e 30 kcal/g proteína. As avaliações antropométricas, alimentares, biquímicas, balanço nitrogenado (BN) e cinética com 15N-glicina foram realizadas no início do estudo, pós D1 (M0) e no último dia das dietas D2 (M1) e D3 (M2). Ao final do estudo (4 semanas) houve aumento significativo na massa muscular (1,63±0,9kg), sem diferença entre D2 e D3. O BN acompanhou o consumo protéico/energético (M0 = -7,8g/dia; M1 = 5,6g/dia e M2 = 16,6g/dia) e a síntese protéica acompanhou o BN, com significância estatística (p<0,05) em relação ao basal (M0) mas semelhante entre D2 e D3 (M1 = 49,8±12,2g N/dia e M2 = 52,5±14,0g N/dia) e sem alteração significativa do catabolismo. Assim, os dados de BN e cinética da 15N-glicina indicam que a ingestão protéica recomendável para esses atletas é superior ao preconizado para sedentários (0,88g/kg) e inferior a 2,5g/kg de peso, sendo no caso, 1,5g de proteína/kg de peso/dia com ajuste do consumo energético para 30 kcal/g de proteína.<br>The effect of increased protein intake on the muscle mass gain, nitrogen balance and 15N-glycine kinetics was studied in six young, healthy subjects practitioners of strength training (> 2 years), without use of anabolic steroids and in agreement with the ethical principles of the research. All athletes received adequate diet (0.88g protein/kg/day) during 2 weeks prior the study (D1), and thereafter with diet providing 1.5g of protein/kg/day and 30kcal/g of protein (D2 diet) for the subsequent 2 weeks. Later on, they all received diet with 2.5g of protein/kg/day (D3 diet) and 30 kcal/g protein for the last two weeks. Body composition, food intake, blood biochemistry, nitrogen balance (NB) and 15N-glycine kinetics were determined at the beginning, after D1 (M0) and in the last days of the D2 (M1) and D3 (M2). The results showed at the end of the study (4 weeks) significant increase in muscle mass (1.63±0.9kg), without difference between D2 and D3. The NB followed the protein/energy consumption (M0 = -7.8g/day; M1 = 5.6g/day and D3 = 16.6g/day), the protein synthesis followed the NB, with M0 < (M1=M2) (M1 = 49.8±12.2g N/day and M2 = 52.5±14.0g N/day). Protein catabolism rate was similarly kept among diets. Thus, the results of the NB and 15N-glycine kinetics indicate that the recommended protein intake for these athletes is higher than the one for sedentary adults (0.88g/kg) and lower than 2.5g/kg, around 1.5g of protein/kg/day, with adjustment of the energy consumption to 30 kcal/g of protein

Pro- and anti-apoptotic roles for IGF-I in TNF-α-induced apoptosis: a MAP kinase mediated mechanism

Objective. The concept of skeletal muscle homeostasis—often viewed as the net balance between two separate processes, namely protein degradation and protein synthesis—are not occurring independently of each other, but are finely co-ordinated by a web of intricate signalling networks. Materials and methods. Using rodent muscle cell lines we have investigated TNF-α/IGF-I interactions, in an attempt to mimic and understand mechanisms underlying the wasting process. Results and conclusion. When myoblast cells are incubated with TNF-α (10 ng ml− 1) maximal damage (21% ± 0.7 myoblast death, p < 0.05) was induced. Co-incubation of TNF-α (10 ng ml− 1) with IGF-I resulted in cell survival (50% reduction in myoblast death, p < 0.05), however, myotube formation was not evident. In contrast, a novel role of IGF-I has been identified whereby co-incubation of muscle cells with IGF-I (1.5 ng ml− 1) and a non-apoptotic dose of TNF-α (1.25 ng ml− 1; sufficient to block differentiation) unexpectedly were shown not to rescue a block on differentiation but to facilitate significant myoblast death (p < 0.05). Interestingly, pre-administration of PD98059, a MAPK signal-blocking agent followed by co-incubation of 1.25 ng ml− 1 TNF-α and 1.5 ng ml− 1 IGF-I, reduced death to baseline levels (p < 0.05). We show for the first time that IGF-I can be apoptotic in the absence of TNF-α-induced cell death