NORMA-Gene: A simple and robust method for qPCR normalization based on target gene data

<p>Abstract</p> <p>Background</p> <p>Normalization of target gene expression, measured by real-time quantitative PCR (qPCR), is a requirement for reducing experimental bias and thereby improving data quality. The currently used normalization approach is based on using one or more reference genes. Yet, this approach extends the experimental work load and suffers from assumptions that may be difficult to meet and to validate.</p> <p>Results</p> <p>We developed a data driven normalization algorithm (NORMA-Gene). An analysis of the performance of NORMA-Gene compared to reference gene normalization on artificially generated data-sets showed that the NORMA-Gene normalization yielded more precise results under a large range of parameters tested. Furthermore, when tested on three very different real qPCR data-sets NORMA-Gene was shown to be best at reducing variance due to experimental bias in all three data-sets compared to normalization based on the use of reference gene(s).</p> <p>Conclusions</p> <p>Here we present the NORMA-Gene algorithm that is applicable to all biological and biomedical qPCR studies, especially those that are based on a limited number of assayed genes. The method is based on a data-driven normalization and is useful for as little as five target genes comprising the data-set. NORMA-Gene does not require the identification and validation of reference genes allowing researchers to focus their efforts on studying target genes of biological relevance.</p

Efficiency and scalability in producing feed from manure using the common housefly

Key-note (Insects to feed the world): Efficiency and scalability in producing feed from manure using the common housefl

Pilot scale production of Hermetia illucens (L.) larvae and frass using former foodstuffs

The food and feed sector requires new sustainable sources of protein and innovative solutions for upcycling of food waste (former foodstuffs), which today is downcycled into energy or even wasted. This study aimed at evaluating the use of former foodstuff waste streams as feed substrate for Hermetia illucens (L.) larvae (black soldier fly larvae, BSFL) under long-term and semi-industrial conditions. Different foodstuff-based mixtures and different stocking BSFL densities were used during 20 batches, and quality and safety assessments were performed on the main outputs, namely BSFL production performance, frass impurities, larval and frass nutrient profiles and heavy metal content. About 1400 kg of former foodstuffs (fresh weight) were used to produce 239 kg BSFL and 230 kg frass. The production of BSFL reared on former foodstuffs was highly efficient, with feed conversion rates (FCR) ranging between 2.3 and 5.5 (dry matter basis). The optimization experiment revealed that former foodstuffs-based mixture and high larval density (10 larvae/cm2) lead to highly efficient (FCR: 2.6) and heavy metal-free production of BSFL and frass. The quality of the derived BSFL meal was high in terms of protein and amino acids. Furthermore, the quality of the technical frass was high in terms of N, P, and K levels and minimal packaging material residuals (&lt;2.65%). This investigation suggests that nutrients in former foodstuffs can be successfully and safely recycled in production of BSFL

Effects of Insect Protein Supplementation during Resistance Training on Changes in Muscle Mass and Strength in Young Men

During prolonged resistance training, protein supplementation is known to promote morphological changes; however, no previous training studies have tested the effect of insect protein isolate in a human trial. The aim of this study was to investigate the potential effect of insect protein as a dietary supplement to increase muscle hypertrophy and strength gains during prolonged resistance training in young men. Eighteen healthy young men performed resistance training four day/week for eight weeks. Subjects were block randomized into two groups consuming either an insect protein isolate or isocaloric carbohydrate supplementation within 1 h after training and pre-sleep on training days. Strength and body composition were measured before and after intervention to detect adaptions to the resistance training. Three-day weighed dietary records were completed before and during intervention. Fat- and bone- free mass (FBFM) improved significantly in both groups (Mean (95% confidence interval (CI))), control group (Con): (2.5 kg (1.5, 3.5) p &lt; 0.01), protein group (Pro): (2.7 kg (1.6, 3.8) p &lt; 0.01) from pre- to post-. Leg and bench press one repetition maximum (1 RM) improved by Con: (42.0 kg (32.0, 52.0) p &lt; 0.01) and (13.8 kg (10.3, 17.2) p &lt; 0.01), Pro: (36.6 kg (27.3, 45.8) p &lt; 0.01) and (8.1 kg (4.5, 11.8) p &lt; 0.01), respectively. No significant differences in body composition and muscle strength improvements were found between groups. In young healthy men, insect protein supplementation did not improve adaptations to eight weeks of resistance training in comparison to carbohydrate supplementation. A high habitual protein intake in both Con and Pro may partly explain our observation of no superior effect of insect protein supplementation

RESEARCH ARTICLE Open Access

NORMA-Gene: A simple and robust method for qPCR normalization based on target gene dat