Electroencephalographic Evidence for Individual Neural Inertia in Mice That Decreases With Time

Previous studies have demonstrated that the brain has an intrinsic resistance to changes in arousal state. This resistance is most easily measured at the population level in the setting of general anesthesia and has been termed neural inertia. To date, no study has attempted to determine neural inertia in individuals. We hypothesize that individuals with markedly increased or decreased neural inertia might be at increased risk for complications related to state transitions, from awareness under anesthesia, to delayed emergence or confusion/impairment after emergence. Hence, an improved theoretical and practical understanding of neural inertia may have the potential to identify individuals at increased risk for these complications. This study was designed to explicitly measure neural inertia in individuals and empirically test the stochastic model of neural inertia using spectral analysis of the murine EEG. EEG was measured after induction of and emergence from isoflurane administered near the EC50 dose for loss of righting in genetically inbred mice on a timescale that minimizes pharmacokinetic confounds. Neural inertia was assessed by employing classifiers constructed using linear discriminant or supervised machine learning methods to determine if features of EEG spectra reliably demonstrate path dependence at steady-state anesthesia. We also report the existence of neural inertia at the individual level, as well as the population level, and that neural inertia decreases over time, providing direct empirical evidence supporting the predictions of the stochastic model of neural inertia.

BXD Recombinant Inbred Mice as a Model to Study Neurotoxicity

BXD recombinant inbred (RI) lines represent a genetic reference population derived from a cross between C57BL/6J mice (B6) and DBA/2J mice (D2), which through meiotic recombination events possesses recombinant chromosomes containing B6 or D2 haplotype segments. The quantitative trait loci (QTLs) are the locations of segregating genetic polymorphisms and are fundamental to understanding genetic diversity in human disease susceptibility and severity. QTL mapping represents the typical approach for identifying naturally occurring polymorphisms that influence complex phenotypes. In this process, genotypic values at markers of known genomic locations are associated with phenotypic values measured in a segregating population. Indeed, BXD RI strains provide a powerful tool to study neurotoxicity induced by different substances. In this review, we describe the use of BXD RI lines to understand the underlying mechanisms of neurotoxicity in response to ethanol and cocaine, as well as metals and pesticide exposures.

Individualized VDJ recombination predisposes the available Ig sequence space

The process of recombination between variable (V), diversity (D), and joining (J) immunoglobulin (Ig) gene segments determines an individual's naive Ig repertoire and, consequently, (auto)antigen recognition. VDJ recombination follows probabilistic rules that can be modeled statistically. So far, it remains unknown whether VDJ recombination rules differ between individuals. If these rules differed, identical (auto)antigen-specific Ig sequences would be generated with individual-specific probabilities, signifying that the available Ig sequence space is individual specific. We devised a sensitivity-tested distance measure that enables inter-individual comparison of VDJ recombination models. We discovered, accounting for several sources of noise as well as allelic variation in Ig sequencing data, that not only unrelated individuals but also human monozygotic twins and even inbred mice possess statistically distinguishable immunoglobulin recombination models. This suggests that, in addition to genetic, there is also nongenetic modulation of VDJ recombination. We demonstrate that population-wide individualized VDJ recombination can result in orders of magnitude of difference in the probability to generate (auto)antigen-specific Ig sequences. Our findings have implications for immune receptor–based individualized medicine approaches relevant to vaccination, infection, and autoimmunity.

Cocaine-induced locomotor activation differs across six sets of inbred mouse substrains

Cocaine use disorders (CUD) are devastating for affected individuals and impose a significant burden on society, but there are currently no FDA-approved therapies. The development of novel and effective treatments has been hindered by substantial gaps in our knowledge about the etiology of these disorders. The risk for developing a CUD is influenced by genetics, the environment and complex interactions between the two. Identifying specific genes and environmental risk factors that increase CUD risk would provide an avenue for the development of novel treatments. Rodent models of addiction-relevant behaviors have been a valuable tool for studying the genetics of response to drugs of abuse. Traditional genetic mapping using genetically and phenotypically divergent inbred mice has been successful in identifying numerous chromosomal regions that influence addiction-relevant behaviors, but these strategies rarely result in identification of the causal gene or genetic variant. To overcome this challenge, reduced complexity crosses (RCC) between closely related inbred mouse substrains have been proposed as a method for rapidly identifying and validating functional variants. The RCC approach is dependent on identifying phenotypic differences between substrains. To date, however, the study of addiction-relevant behaviors has been limited to very few sets of substrains, mostly comprising the C57BL/6 lineage. The present study expands upon the current literature to assess cocaine-induced locomotor activation in 20 inbred mouse substrains representing six inbred strain lineages (A/J, BALB/c, FVB/N, C3H/He, DBA/2 and NOD) that were either bred in-house or supplied directly by a commercial vendor. To our knowledge, we are the first to identify significant differences in cocaine-induced locomotor response in several of these inbred substrains. The identification of substrain differences allows for the initiation of RCC populations to more rapidly identify specific genetic variants associated with acute cocaine response. The observation of behavioral profiles that differ between mice generated in-house and those that are vendor-supplied also presents an opportunity to investigate the influence of environmental factors on cocaine-induced locomotor activity.

The new attenuated strain of Mycobacterium tuberculosis BN. Characteristics and vaccine properties

The objective of the study: to obtain a live attenuated strain and investigate its properties by multiple cultures of the virulent strain of Mycobacterium tuberculosis H37Rv.Subjects and Methods. The original virulent strain H37Rv was subcultured 70 times in 7H9 liquid medium. Genetic properties of the new strain, degree of avirulence, and vaccine properties were studied.Results. Mycobacteria of the new strain MtbBN lost their virulence to inbred mice. Eight mutations were identified by whole genome sequencing: single nucleotide insertions and deletions (in/del) distinguishing the MtbBN and H37Rv strains. The MtbBN strain demonstrated vaccine potential at the BCG level. Additionally, in some genetic models, the attenuated strain was highly effective in protecting inbred mice when infected with Mtb H37Rv as opposed to BCG.

Distinct Morphological and Behavioural Alterations in ENU-Induced Heterozygous Trpc7K810Stop Mutant Mice

Trpc7 (transient receptor potential cation channel, subfamily C, member 7; 862 amino acids) knockout mice are described showing no clear phenotypic alterations, therefore, the functional relevance of the gene remains unclear. A complementary approach for the functional analysis of a given gene is the examination of individuals harbouring a mutant allele of the gene. In the phenotype-driven Munich ENU mouse mutagenesis project, a high number of phenotypic parameters was used for establishing novel mouse models on the genetic background of C3H inbred mice. The phenotypically dominant mutant line SMA002 was established and further examined. Analysis of the causative mutation as well as the phenotypic characterization of the mutant line were carried out. The causative mutation was detected in the gene Trpc7 which leads to the production of a truncated protein due to the novel stop codon at amino acid position 810 thereby affecting the highly conserved cytoplasmic C terminus of the protein. Trpc7 heterozygous mutant mice of both sexes were viable and fertile, but showed distinct morphological and behavioural alterations which is in contrast to the published phenotype of Trpc7 knockout mice. Thus, the Trpc7K810Stop mutation leads to a dominant negative effect of the mutant protein.

A Diversity Outbred F1 mouse model identifies host-intrinsic genetic regulators of response to immune checkpoint inhibitors

: Immune checkpoint inhibitors (ICI) have improved outcomes for a variety of malignancies; however, many patients fail to benefit. While tumor-intrinsic mechanisms are likely involved in therapy resistance, it is unclear to what extent host genetic background influences response. To investigate this, we utilized the Diversity Outbred (DO) and Collaborative Cross (CC) mouse models. DO mice are an outbred stock generated by crossbreeding 8 inbred founder strains, and CC mice are recombinant inbred mice generated from the same 8 founders. We generated 207 DOB6F1 mice representing 48 DO Dams and demonstrated that these mice reliably accept the C57BL/6 syngeneic B16F0 tumor and that host genetic background influences response to ICI. Genetic linkage analysis from 142 mice identified multiple regions including one within chromosome 13 that associated with therapeutic response. We utilized 6 CC strains bearing the positive (NZO) or negative (C57BL/6) driver genotype in this locus. We found that 2/3 of predicted responder CCB6F1 crosses show reproducible ICI response. The chromosome 13 locus contains the murine prolactin family, which is a known immunomodulating cytokine associated with various autoimmune disorders. To directly test whether prolactin influences ICI response rates, we implanted inbred C57BL/6 mice with subcutaneous slow-release prolactin pellets to induce mild hyperprolactinemia. Prolactin augmented ICI response against B16F0, with 5/8 mice exhibiting slowed tumor growth relative to controls. This study highlights the role of host genetics in ICI response and supports the use of F1 crosses in the DO and CC mouse populations as powerful cancer immunotherapy models.

Terc Gene Cluster Variants Predict Liver Telomere Length in Mice

Variants in a gene cluster upstream-adjacent to TERC on human chromosome 3, which includes genes APRM, LRRC31, LRRC34 and MYNN, have been associated with telomere length in several human populations. Currently, the mechanism by which variants in the TERC gene cluster influence telomere length in humans is unknown. Given the proximity between the TERC gene cluster and TERC (~0.05 Mb) in humans, it is speculated that cluster variants are in linkage disequilibrium with a TERC causal variant. In mice, the Terc gene/Terc gene cluster are also located on chromosome 3; however, the Terc gene cluster is located distantly downstream of Terc (~60 Mb). Here, we initially aim to investigate the interactions between genotype and nicotine exposure on absolute liver telomere length (aTL) in a panel of eight inbred mouse strains. Although we found no significant impact of nicotine on liver aTL, this first experiment identified candidate single nucleotide polymorphisms (SNPs) in the murine Terc gene cluster (within genes Lrrc31, Lrriq4 and Mynn) co-varying with aTL in our panel. In a second experiment, we tested the association of these Terc gene cluster variants with liver aTL in an independent panel of eight inbred mice selected based on candidate SNP genotype. This supported our initial finding that Terc gene cluster polymorphisms impact aTL in mice, consistent with data in human populations. This provides support for mice as a model for telomere dynamics, especially for studying mechanisms underlying the association between Terc cluster variants and telomere length. Finally, these data suggest that mechanisms independent of linkage disequilibrium between the Terc/TERC gene cluster and the Terc/TERC gene mediate the cluster’s regulation of telomere length.

Comparative analysis of the phytochemical and antibacterial activity of the root extracts of Euphorbia heterophylla and Vitellaria paradoxa

Background: Over time, herbal plants and their various components have been major sources of therapeutic medicine for man. A comparative study was carried out to determine the phytochemical components and antibacterial activities of the different crude extracts of Euphorbia heterophylla and Vitellaria paradoxa roots on four enteric bacteria; Salmonella typhi, Shigella flexneri, Escherichia coli and Proteus vulgaris.Methodology: Root samples of E. heterophylla and V. paradoxa were collected, washed, air dried and processed to fine powder in the microbiology laboratory of Federal University of Technology, Minna, Nigeria. Crude extract of the root samples was done by the cold maceration technique using four solvents (chloroform, methanol, petroleum ether and water). Phytochemical analysis of the extracts was done using previously described technique, and in vitro antibacterial activities of different concentrations of the extracts (50-200 mg/ml) and a standard antibiotic (ciprofloxacin) were tested on four enteric bacteria (S. typhi, S. flexneri, E. coli, P. vulgaris) by the agar diffusion test. In vivo antibacterial activities of the two plants were also tested by daily oral administration of 2000 mg/kg bodyweight (for 7 days) of each extract on inbred mice infected through intraperitoneal inoculation of an infective dose of each of the four enteric bacteria. Data were computed as mean ± standard error and analysed by the Statistical Analysis System (SAS) version 9.4. Associations between variables were determined using analysis of variance (ANOVA), with p < 0.05 considered as significant value.Results: Phytochemical analysis of the crude extracts of both plants revealed the presence of cardiac glycosides, saponins, alkaloids, flavonoids, and tannins but V. paradoxa contain more carbohydrates and starch, and less phlobatannins, compared to E. heterophylla. In vitro assay showed dose-dependent antibacterial activity of the methanol, aqueous and chloroform (but not petroleum ether) extracts of the two plant roots. The in vitro antibacterial activities of the different extracts of V. paradoxica extracts were significantly higher (higher mean diameters of inhibition zones) than those of E. heterophylla (p<0.05), and methanol extracts gave the highest antibacterial effects. However, the root extract of E. heterophylla gave a higher antibacterial activity with the in vivo assay on inbred mice than V. paradoxa, and methanol extracts of the two plant extracts gave the highest in vivo activity, followed by aqueous extract and least activity was obtained with the chloroform extract.Conclusion: Crude extracts of E. heterophylla and V. paradoxa roots produce antibacterial activity against enteric Gram-negative bacteria pathogens involved in diarrhoea illnesses. Further researches should be directed towards isolation and characterization of the active compounds in the crude extracts. French title: Analyse comparative de l'activité phytochimique et antibactérienne des extraits de racines d'Euphorbia heterophylla et de Vitellaria paradoxa Contexte: Au fil du temps, les plantes médicinales et leurs divers composants ont été une source majeure de médecine thérapeutique pour l'homme. Une étude comparative a été réalisée pour déterminer les composants phytochimiques et les activités antibactériennes des différents extraits bruts de racines d'Euphorbia heterophylla et de Vitellaria paradoxa sur quatre bactéries entériques; Salmonella typhi, Shigella flexneri, Escherichia coli et Proteus vulgaris. Méthodologie: Des échantillons de racines d'E. heterophylla et de V. paradoxa ont été collectés, lavés, séchés à l'air et transformés en poudre fine dans le laboratoire de microbiologie de l'Université fédérale de technologie, Minna, Nigéria. L'extraction brute des échantillons de racines a été réalisée par la technique de macération à froid en utilisant quatre solvants (chloroforme, méthanol, éther de pétrole et eau). L'analyse phytochimique des extraits a été effectuée en utilisant la technique décrite précédemment, et les activités antibactériennes in vitro de différentes concentrations des extraits (50-200 mg/ml) et d'un antibiotique standard (ciprofloxacine) ont été testées sur quatre bactéries entériques (S. typhi, S. flexneri, E. coli, P. vulgaris) par le test de diffusion sur gélose. Les activités antibactériennes in vivo des deux plantes ont également été testées par administration orale quotidienne de 2000 mg/kg de poids corporel (pendant 7 jours) de chaque extrait sur des souris consanguines infectées par inoculation intrapéritonéale d'une dose infectieuse de chacune des quatre bactéries entériques. Les données ont été calculées en tant que moyenne ± erreur standard et analysées par le système d'analyse statistique (SAS) version 9.4. Les associations entre les variables ont été déterminées à l'aide d'une analyse de variance (ANOVA), avec p < 0,05 considéré comme une valeur significative. Résultats: L'analyse phytochimique des extraits bruts des deux plantes a révélé la présence de glycosides cardiaques, de saponines, d'alcaloïdes, de flavonoïdes et de tanins mais V. paradoxa contient plus de glucides et d'amidon, et moins de phlobatannins, par rapport à E. heterophylla. Un essai in vitro a montré une activité antibactérienne dose-dépendante des extraits au méthanol, aqueux et au chloroforme (mais pas à l'éther de pétrole) des deux racines des plantes. Les activités antibactériennes in vitro des différents extraits d'extraits de V. paradoxica étaient significativement plus élevées (diamètres moyens des zones d'inhibition plus élevés) que celles d'E. heterophylla (p<0,05), et les extraits au méthanol ont donné les effets antibactériens les plus élevés. Cependant, l'extrait de racine d'E. heterophylla a donné une activité antibactérienne plus élevée avec le test in vivo sur des souris consanguines que V. paradoxa, et les extraits au méthanol des deux extraits de plantes ont donné l'activité in vivo la plus élevée, suivie par l'extrait aqueux et l'activité la plus faible a été obtenu avec l'extrait chloroformique. Conclusion: Des extraits bruts de racines d'E. heterophylla et de V. paradoxa produisent une activité antibactérienne contre les bactéries pathogènes entériques à Gram négatif impliquées dans les maladies diarrhéiques. D'autres recherches devraient être dirigées vers l'isolement et la caractérisation des composés actifs dans les extraits bruts.