We investigated the effects of plant density on the welfare of African catfish, Clarias gariepinus, in coupled aquaponics over 85 days. The moderate density (mpd) of basil, Ocimum basilicum, was compared with the high density (hpd) and control (n = 0). The behavior was analyzed by visual and video observations, and after the application of induced stressors, skin injuries, blood glucose, lactate, and plasma cortisol responses were considered. The hpd fish showed the least activity (control: visual 77.8%, video 81.6%; mpd: 74.6%, 82.6%; hpd: 63.2% [p < 0.05], 78.8%). High agonistic behavior (control: 5, 131; mpd: 4, 57; hpd: 1, 45) and the highest number of injuries (control: 3.9; mpd: 2.9; hpd: 3.4) were observed in the control. Glucose and lactate levels did not differ significantly (control: 5.5, 2.6 mmol/L; mpd: 5.6, 2.7 mmol/L; hpd: 5.3, 2.6 mmol/L); however, cortisol levels did (control: 18.8 ng/mL, mpd: 19.9 ng/mL, hpd: 25.8 ng/mL). pH adjustment led to additional stress, resulting in temporal cortisol alterations. While in the control and mpd, low cortisol levels were followed by acute responses and downregulation, the hpd fish showed prior elevation and lagged an acute response. However, comparing injuries and behavioral patterns with control, aquaponics with high basil density influenced African catfish positively.
Background Next Generation Sequencing (NGS) is the fundament of various studies, providing insights into questions from biology and medicine. Nevertheless, integrating data from different experimental backgrounds can introduce strong biases. In order to methodically investigate the magnitude of systematic errors in single nucleotide variant calls, we performed a cross-sectional observational study on a genomic cohort of 99 subjects each sequenced via (i) Illumina HiSeq X, (ii) Illumina HiSeq, and (iii) Complete Genomics and processed with the respective bioinformatic pipeline. We also repeated variant calling for the Illumina cohorts with GATK, which allowed us to investigate the effect of the bioinformatics analysis strategy separately from the sequencing platform’s impact. Results The number of detected variants/variant classes per individual was highly dependent on the experimental setup. We observed a statistically significant overrepresentation of variants uniquely called by a single setup, indicating potential systematic biases. Insertion/deletion polymorphisms (indels) were associated with decreased concordance compared to single nucleotide polymorphisms (SNPs). The discrepancies in indel absolute numbers were particularly prominent in introns, Alu elements, simple repeats, and regions with medium GC content. Notably, reprocessing sequencing data following the best practice recommendations of GATK considerably improved concordance between the respective setups. Conclusion We provide empirical evidence of systematic heterogeneity in variant calls between alternative experimental and data analysis setups. Furthermore, our results demonstrate the benefit of reprocessing genomic data with harmonized pipelines when integrating data from different studies.
Voltage-gated calcium channels (VGCCs) represent key regulators of the calcium influx through the plasma membrane of excitable cells, like neurons. Activated by the depolarization of the membrane, the opening of VGCCs induces very transient and local changes in the intracellular calcium concentration, known as calcium nanodomains, that in turn trigger calcium-dependent signaling cascades and the release of chemical neurotransmitters. Based on their central importance as concierges of excitation-secretion coupling and therefore neuronal communication, VGCCs have been studied in multiple aspects of neuronal function and malfunction. However, studies on molecular interaction partners and recent progress in omics technologies have extended the actual concept of these molecules. With this review, we want to illustrate some new perspectives of VGCCs reaching beyond their function as calcium-permeable pores in the plasma membrane. Therefore, we will discuss the relevance of VGCCs as voltage sensors in functional complexes with ryanodine receptors, channel-independent actions of auxiliary VGCC subunits, and provide an insight into how VGCCs even directly participate in gene regulation. Furthermore, we will illustrate how structural changes in the intracellular C-terminus of VGCCs generated by alternative splicing events might not only affect the biophysical channel characteristics but rather determine their molecular environment and downstream signaling pathways.
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