Genetic Mouse Models of Liver Disease: Potential Roles of Zhx2 (Afr1) and Afr2 in Damage and Regeneration

The liver is the largest internal organ in mammals and responsible for carrying out various processes, including lipid and carbohydrate metabolism, detoxification of chemicals, and production of serum proteins. Liver damage, which can be caused by a variety of agents including viral infection, environmental toxins, alcohol and excessive dietary fats, can cause dysregulation of these critical functions, leading to worsening liver pathophysiology and impacting health. However, the liver has the remarkable ability to regenerate when damaged. Hepatocytes, which comprise a majority of liver cells, are relatively quiescent under healthy conditions. Upon injury, remaining hepatocytes can proliferate to recover from liver damage while performing their metabolic functions. Unfortunately, persistent injury can progressively lead to hepatitis, fibrosis, and ultimately end-stage liver disease, cirrhosis, and liver cancer. Overall, liver disease is the fifth leading cause of death worldwide and a growing healthcare problem in the United States and other western countries. A better understanding of liver regeneration could be used clinically to treat a wide range of liver diseases. Studies in different mouse strains have provided genetic models to understand aspects of liver gene regulation and liver disease. Two regulators of the liver-derived serum protein alpha-fetoprotein (AFP), alpha-fetoprotein regulator 1 and 2 (Afr1 and Afr2), have been implicated to play a role in liver disease and regeneration. AFP is normally expressed in the fetal liver, silenced at birth, and reactivated in liver regeneration and liver cancer. Based on the unusually high AFP expression in adult BALB/cJ mice, our lab identified Zinc Fingers and Homeoboxes 2 (Zhx2) as the gene responsible for the Afr1 trait. BALB/cJ mice contain a natural mutation in the Zhx2 gene. Additional studies have shown that Zhx2 is involved in several liver diseases, including diet-induced fatty liver disease and hepatocellular carcinoma. In addition to these traits, BALB/cJ mice have been shown to have increased liver fibrosis after chronic treatment with the hepatotoxin carbon tetrachloride (CCl4). The locus responsible for this trait, called Hepatic fibrosis 1 (Hfib1), was mapped to the same region of Chromosome 15 (Chr15) as Zhx2, but the Hfib1 gene has not been identified. During liver regeneration after acute treatment with CCl4, it was found that liver AFP mRNA levels were much higher in C3H/HeJ mice than in C57BL/6J mice. The locus that controls this strain-specific difference in AFP reactivation was called Afr2; C3H/HeJ and C57BL/6 mice are thought to contain the Afr2a and Afr2b alleles, respectively. The Afr2 locus has been mapped to Chr2, but the Afr2 gene has not been identified. This dissertation tested the hypothesis that Zhx2 is responsible for the Hfib1 trait in BALB/cJ mice. Using BALB/cJ mice and C57BL/6J mice with a targeted mutation in the Zhx2 gene, my data indicates that Zhx2 is not responsible for the Hfib1 trait. Mice with low Zhx2 expression did not have more significant inflammation, liver damage, or fibrosis than mice with wild-type Zhx2 levels. These data suggest that another gene, presumably within the same region as Zhx2, is responsible for the Hfib1 phenotype in BALB/cJ mice. I also analyzed the Afr2 trait across several different strains of mice. My studies indicate that the strains 129X1/SvJ, C3H/HeJ, and DBA/2J contain the Afr2a allele, whereas mice in the C57 lineage (C57BL/6J, C57BL/6N, C57BL/10) contain the Afr2b allele. I also demonstrate that F1 offspring of 129X1/SvJ mice and C57BL/6J mice display an intermediate AFP reactivation phenotype, suggesting that the Afr2a and Afr2b alleles are co-dominant. These data provide the framework for future studies to identify the Afr2 gene. Taken together, my results indicate that regulators of gene expression within the liver, as defined by differences within mouse strains, can provide insight into liver disease and regeneration

Negative Findings Report Cultural Resources Survey Fabens Lease 13031, University Lands El Paso County, Texas

On July 23, 2020, TAS Inc. conducted a cultural resource survey of a 10- acre tract intended to host a truck stop on the northwest corner of IH10 and FM 793 two miles northeast of the town of Fabens, in El Paso County, Texas. The project was sponsored by Ya Rehman, Ent, Inc. and was authorized by Texas Antiquities Permit 9518 with Jeff Turpin acting as Principal Investigator. The tract is a southwest-trending dune between two arroyos. The pavement and construction debris from an abandoned gasoline station occupy the southwestern corner. The current survey and shovel test regime found the remains of the gas station, but no new archeological evidence of any kind was discovered Consequently cultural resources present no obstacle to construction of the proposed truck stop, and justify a finding of “no effect”. However, if cultural material is encountered during the course of construction, work in that area should cease and the various regulatory agencies should be advised

Negative Findings Report Cultural Resource Survey Fabens Lease 1301, University Lands El Paso County, Texas

On July 23, 2020, TAS Inc. conducted a cultural resource survey of a 10- acre tract intended to host a truck stop on the northwest corner of IH10 and FM 793 two miles northeast of the town of Fabens, in El Paso County, Texas. The project was sponsored by Ya Rehman, Ent, Inc. and was authorized by Texas Antiquities Permit 9518 with Jeff Turpin acting as Principal Investigator. The tract is a southwest-trending dune between two arroyos. The pavement and construction debris from an abandoned gasoline station occupy the southwestern corner. The current survey and shovel test regime found the remains of the gas station, but no new archeological evidence of any kind was discovered Consequently cultural resources present no obstacle to construction of the proposed truck stop, and justify a finding of “no effect”. However, if cultural material is encountered during the course of construction, work in that area should cease and the various regulatory agencies should be advised

A successful Iowa shed roof poultry house

After putting it to practical test on the Iowa State College poultry farm, the Iowa Agricultural Experiment Station recommends the permanent shed roof poultry house described in this bulletin. This house was planned and built by the Poultry and Agricultural Engineering sections of the station for housing 75 or more laying hens kept under Iowa conditions. It has given good results in practical use at the poultry farm and on this basis it is recommended to those who have need of a poultry house of this type

Improving welfare and production in the peri-weaning period: Effects of co-mingling and intermittent suckling on the stress response, performance, behaviour, and gastrointestinal tract carbohydrate absorption in young pigs

We investigated the effect of different pre-weaning interventions on performance, aspects of behaviour, and selected neuroendocrine, inflammatory and immune indices in 593 weanling pigs (59 litters, weaning age 22 ± 1.7). Measurements were taken at various time points two weeks before and after weaning. Sugar absorption tests (20% mannitol and 20% galactose solutions) were used to assess gastrointestinal tract (GIT) absorptive capacity. One week before weaning, litters were either co-mingled (CoM) for 8 h daily with another litter or not co-mingled (NoCoM). Half of the litters were also subjected to intermittent suckling (IS) involving separation from their sow for 8 h daily and the other half remained with their sow (NoIS). Hence, four treatments were produced in a 2×2 factorial design; (1) CoM IS (n = 16 litters), (2) CoM NoIS (n = 14 litters), (3) NoCoM IS (n = 16 litters), (4) NoCoM NoIS (n = 13 litters). Measurements are compared within each of the main effects (CoM or IS) unless otherwise stated. Acute weaning stress was evidenced by increases in cortisol, haptoglobin and N: L ratios when data were combined (P < 0.001). However, there were tendencies (P < 0.1) for lower cortisol in IS pigs and higher N: L ratios in CoM pigs at weaning. While CoM did not affect performance before weaning, growth (P < 0.05), feed intake (P < 0.05) and body weight (P < 0.05) were reduced in CoM pigs 7-14 days after weaning. One week of IS before weaning improved feed intake before weaning (P < 0.01), resulting in better growth (P = 0.01) and a tendency (P < 0.1) for a higher feed intake 2-7 days after weaning. Co-mingled piglets had more scratches 4 days before weaning (P < 0.001), but tended to have fewer scratches 2 days after weaning (P < 0.1). Pigs exposed to either IS or CoM displayed more sleeping behaviour the day after weaning (P < 0.01 and P < 0.001). A higher mannitol absorption was evident in CoM NoIS pigs 3 days after weaning (P < 0.01), and galactose absorption was reduced in IS pigs 3 days before weaning (P < 0.05) and tended to be reduced 3 days after weaning (P < 0.1), likely reflecting a GIT adaptive response. Overall, despite improvements in mannitol absorption and behaviour, there was no beneficial effect of CoM on performance after weaning. Alternatively, IS improved behaviour and performance during the first week after weaning

Intermittent suckling with or without co-mingling of non-littermate piglets before weaning improves piglet performance in the immediate post-weaning period when compared with conventional weaning

Background: In this experiment, intermittent suckling (IS) with or without the co-mingling (CoM) of piglets was studied as a method to stimulate solid feed intake and reduce post-weaning stress. Methods: Three weaning regimes using 30 multiparous sows were compared: (1) conventional weaning (CW) (n = 10 litters), where piglets had continuous access to the sow until weaning (d 0, farrowing = d −25 relative to weaning); (2) intermittent suckling (IS) (n = 10 litters), where piglets were separated from the sow for 8 h/d starting at d −7 (relative to weaning); and (3) intermittent suckling with co-mingling (ISCo) (n = 10 litters) where IS started at d −7 and two litters were housed together during separation and then returned to their original sow. Ad libitum creep feed was available from d −17. At weaning pigs were housed in pens of 11 pigs, 27 pens in total. The ISCo treatment was divided in half to examine effects of different mixing strategies after weaning. Half of the ISCo litters were kept in familiar groups (ISCoF, familiar, n = 4) and the other half were mixed within treatment resulting in groups of unfamiliar pigs (ISCoNF, not familiar, n = 5), the same as IS (n = 9) and CW (n = 9) treatments. Results: The ISCo piglets ate more creep feed in the week before weaning (P < 0.01), but also showed more aggressive and manipulative behaviour on first day of CoM compared with CW piglets (P < 0.05). IS with or without CoM increased exploratory and play behaviour on the first day of treatment intervention (P < 0.001) and increased sleeping behaviour on the last day of treatment intervention compared with CW (P < 0.001). Mixing strategy at weaning had an effect on performance data with the highest growth and feed intake seen in ISCoF pigs 2 to 8 d after weaning (P <0.001). IS and ISCoNF pigs also grew faster and ate more than CW pigs 2 to 8 d after weaning (P < 0.001). Post-weaning injury scores suggested reduced aggression in ISCo as evidenced by reduced redness (skin irritation) (P < 0.05), and a tendency for ISCo to have less scratches than CW (P < 0.1). The IS pigs slept the most and displayed less manipulative behaviours on the day of weaning and plasma haptoglobin levels remained low in IS pigs after weaning (P ≤ 0.01). Conclusions: Both intermittent suckling techniques improved production indices in the immediate post-weaning period. However, the addition of co-mingling before weaning in combination with grouping familiar pigs together after weaning improved performance in an additive manner

Oligomers, organosulfates, and nitrooxy organosulfates in rainwater identified by ultra-high resolution electrospray ionization FT-ICR mass spectrometry

Wet deposition is an important removal mechanism for atmospheric organic matter, and a potentially important input for receiving ecosystems, yet less than 50% of rainwater organic matter is considered chemically characterized. Precipitation samples collected in New Jersey, USA, were analyzed by negative ion ultra-high resolution electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Elemental compositions of 552 unique molecular species were determined in the mass range 50–500 Da in the rainwater. Four main groups of organic compounds were identified: compounds containing carbon, hydrogen, and oxygen (CHO) only, sulfur (S) containing CHOS compounds, nitrogen (N) containing CHON compounds, and S- and N- containing CHONS compounds. Organic acids commonly identified in precipitation were detected in the rainwater. Within the four main groups of compounds detected in the rainwater, oligomers, organosulfates, and nitrooxy-organosulfates were assigned based on elemental formula comparisons. The majority of the compounds identified are products of atmospheric reactions and are known contributors to secondary organic aerosol (SOA) formed from gas phase, aerosol phase, and in-cloud reactions in the atmosphere. It is suggested that the large uncharacterized component of SOA is the main contributor to the large uncharacterized component of rainwater organic matter

Intermittent suckling in combination with an older weaning age improves growth, feed intake and aspects of gastrointestinal tract carbohydrate absorption in pigs after weaning

This study tested the hypothesis that intermittent suckling (IS) with or without an older weaning age would improve post-weaning gastrointestinal tract (GIT) carbohydrate absorptive capacity in pigs while reducing post-weaning stress and aspects of the inflammatory response. Three weaning regimes using primiparous sows were compared: (1) conventional weaning (CW28) (n = 22), where piglets were weaned conventionally at day 28; (2) IS28 (n = 21), where IS started at day 21 until weaning at day 28; and (3) IS35 (n = 21), where IS started at day 28 until weaning at day 35. Sugar absorption tests (10% mannitol or 10% galactose) were used to measure GIT absorptive capacity. All measured parameters were compared in relation to weaning across treatments (i.e., different physiological ages were compared). The IS35 pigs grew fastest in the 12 days after weaning (p < 0.01) and had the highest solid feed intake before and after weaning (p < 0.05). Irrespective of treatment, pre-weaning mannitol levels were higher than post-weaning levels (p < 0.01), whereas post-weaning galactose levels were highest in IS35 pigs (p < 0.01). Cytokine data did not show any treatment effects. In conclusion, these data suggest that IS in combination with an older weaning age (day 35) improved post-weaning adaptation as evidenced by improvements in performance measures and galactose absorption. However, IS28 did not improve post-weaning performance