ProThera Biologics (United States)

Inter-alpha Inhibitor Proteins (IAIP) are serine protease inhibitors and a reliable inflammatory biomarker. We have demonstrated that IAIP levels decrease during sepsis in humans and animal models. Currently, enzyme-linked immunosorbent assay (ELISA) is the standard procedure to measure IAIP levels. We developed a lateral flow immunoassay (LFIA) that allows rapid, point of care detection. We compared IAIP levels in infants with sepsis and/or necrotizing enterocolitis (NEC) by ELISA and LFIA in a multi-center, cross-sectional study. Blood samples were collected from 47 infants with sepsis, 31 sepsis case controls, 52 gestational age (GA)-matched controls and 10 infants with culture-negative sepsis (“clinical sepsis”). We also collected samples from 17 infants with NEC, 7 NEC case controls and 15 GA controls. IAIP levels at presentation of acute events and over the next 72 hours were significantly reduced in infants with sepsis, NEC and culture negative sepsis when compared to controls. IAIP levels did not differ in patients with sepsis or culture negative sepsis. IAIP levels measured by ELISA and LFIA were highly correlated (R2 = 0.9326) and both showed reliable detection of neonatal sepsis, NEC and culture negative sepsis. IAIP levels were 80.0% sensitive and 92.3% specific using LFIA for the detection of neonatal sepsis. For detection of NEC, IAIP levels were 84.6% sensitive and 86.7% specific.

Mammalian DNA replication employs several RecQ DNA helicases to orchestrate the faithful duplication of genetic information. Helicase function is often coupled to the activity of specific nucleases, but how helicase and nuclease activities are co-directed is unclear. Here we identify the inactive ubiquitin-specific protease, USP50, as a ubiquitin-binding and chromatin-associated protein required for ongoing replication, fork restart, telomere maintenance and cellular survival during replicative stress. USP50 supports WRN:FEN1 at stalled replication forks, suppresses MUS81-dependent fork collapse and restricts double-strand DNA breaks at GC-rich sequences. Surprisingly we find that cells depleted for USP50 and recovering from a replication block exhibit increased DNA2 and RECQL4 foci and that the defects in ongoing replication, poor fork restart and increased fork collapse seen in these cells are mediated by DNA2, RECQL4 and RECQL5. These data define a novel ubiquitin-dependent pathway that promotes the balance of helicase: nuclease use at ongoing and stalled replication forks.

Plant introductions outside their native ranges by humans have led to substantial ecological consequences. While we have gained considerable knowledge about intercontinental introductions, the distribution and determinants of intracontinental aliens remain poorly understood. Here, we studied naturalized (i.e., self-sustaining) intracontinental aliens using native and alien floras of 243 mainland regions in North America, South America, Europe, and Australia. We revealed that 4510 plant species had intracontinental origins, accounting for 3.9% of all plant species and 56.7% of all naturalized species in these continents. In North America and Europe, the numbers of intracontinental aliens peaked at mid-latitudes, while the proportion peaked at high latitudes in Europe. Notably, we found predominant poleward naturalization, primarily due to larger native species pools in low-latitudes. Geographic and climatic distances constrained the naturalization of intracontinental aliens in Australia, Europe, and North America, but not in South America. These findings suggest that poleward naturalizations will accelerate, as high latitudes become suitable for more plant species due to climate change.

A taxonomic treatment is presented for the violets (Violaceae) confirmed in the range described in the second edition of Gleason and Cronquist's Manual of Vascular Plants of Northeastern United States and Adjacent Canada, in advance of the new treatment of this manual that is in preparation. Recent field, herbarium, and laboratory investigations of eastern North America's violet flora by the author and students have reexamined eastern North American violet taxa and utilized multiple sources of evidence filtered through modern species concepts to delimit taxa and assign appropriate taxonomic rank. Results have led to the recognition of 58 native and eight introduced species, one subspecies, one variety and one form, 10 informal variants, and 113 wild hybrids in our region, representing the genera Cubelium, Pombalia, and Viola. In Viola, two new species are described and three new combinations are made (one to species rank, one to subspecies, and one to forma), and eight names are typified. Viola taxa are distributed across five sections and nine subsections and are representative of most of the infrageneric taxonomic diversity in North America overall. Anomalous variants in Viola emarginata, Viola palmata, Viola septentrionalis, and Viola sororia currently lack sufficient information for taxonomic decisions but are presented to encourage study, collection, and documentation. Six potential taxa occur adjacent to or very near our region and may eventually be discovered in our area; they are included in the keys and briefly described with figures. The treatment includes introductory text, dichotomous keys, synonymy with some type information, detailed descriptions, and other information, as well as color figures of habit and plant structures and US county-level geographic distribution maps for nearly all included taxa.

Hypoxia–ischemia (HI)-related brain injury is an important cause of morbidity and long-standing disability in newborns. We have previously shown that human plasma-derived inter-alpha inhibitor proteins (hIAIPs) attenuate HI-related brain injury in neonatal rats. The optimal dose of hIAIPs for their neuroprotective effects and improvement in behavioral outcomes remains to be determined. We examined the efficacy of 30, 60, or 90 mg/kg of hIAIPs administered to neonatal rats after exposure to HI for 2 h. Postnatal day 7 (P7) Wistar rats were exposed to either sham-surgery or unilateral HI (right carotid artery ligation, 2 h of 8% O2) brain injury. A placebo, 30, 60, or 90 mg/kg of hIAIPs were injected intraperitoneally at 0, 24 and 48 h after HI (n = 9–10/sex). We carried out the following behavioral analyses: P8 (righting reflex), P9 (negative geotaxis) and P10 (open-field task). Rats were humanely killed on P10 and their brains were stained with cresyl violet. Male extension/contraction responses and female righting reflex times were higher in the HI placebo groups than the sham groups. Female open-field exploration was lower in the HI placebo group than the sham group. hIAIPs attenuated these behavioral deficits. However, the magnitude of the responses did not vary by hIAIP dose. hIAIPs reduced male brain infarct volumes in a manner that correlated with improved behavioral outcomes. Increasing the hIAIP dose from 30 to 90 mg/kg did not further accentuate the hIAIP-related decreases in infarct volumes. We conclude that larger doses of hIAIPs did not provide additional benefits over the 30 mg/kg dose for behavior tasks or reductions in infarct volumes in neonatal rats after exposure to severe HI.

Inter-alpha Inhibitor Proteins (IAIPs) are key immunomodulatory molecules. Endogenous IAIPs are present in human, rodent, and sheep brains, and are variably localized to the cytoplasm and nuclei at multiple developmental stages. We have previously reported that ischemia-reperfusion (I/R) reduces IAIP concentrations in the fetal sheep brain. In this study, we examined the effect of I/R on total, cytoplasmic, and nuclear expression of IAIPs in neurons (NeuN+), microglia (Iba1+), oligodendrocytes (Olig2+) and proliferating cells (Ki67+), and their co-localization with histones and the endoplasmic reticulum in fetal brain cells. At 128 days of gestation, fetal sheep were exposed to Sham (n = 6) or I/R induced by cerebral ischemia for 30 min with reperfusion for 7 days (n = 5). Although I/R did not change the total number of IAIP+ cells in the cerebral cortex or white matter, cells with IAIP+ cytoplasm decreased, whereas cells with IAIP+ nuclei increased in the cortex. I/R reduced total neuronal number but did not change the IAIP+ neuronal number. The proportion of cytoplasmic IAIP+ neurons was reduced, but there was no change in the number of nuclear IAIP+ neurons. I/R increased the number of microglia and decreased the total numbers of IAIP+ microglia and nuclear IAIP+ microglia, but not the number of cytoplasmic IAIP+ microglia. I/R was associated with reduced numbers of oligodendrocytes and increased proliferating cells, without changes in the subcellular IAIP localization. IAIPs co-localized with the endoplasmic reticulum and histones. In conclusion, I/R alters the subcellular localization of IAIPs in cortical neurons and microglia but not in oligodendrocytes or proliferating cells. Taken together with the known neuroprotective effects of exogenous IAIPs, we speculate that endogenous IAIPs may play a role during recovery from I/R.

Circulating levels of inter-alpha inhibitor proteins change dramatically in acute inflammatory disorders, which suggest an important contribution to the immunomodulatory system. Human blood-derived inter-alpha inhibitor proteins are neuroprotective and improve survival of neonatal mice exposed to lipopolysaccharide. Lipopolysaccharide augments inflammatory conditions and disrupts the blood–brain barrier. There is a paucity of therapeutic strategies to treat blood–brain barrier dysfunction, and the neuroprotective effects of human blood-derived inter-alpha inhibitor proteins are not fully understood. To examine the therapeutic potential of inter-alpha inhibitor proteins, we administered human blood-derived inter-alpha inhibitor proteins to male and female CD-1 mice after lipopolysaccharide exposure and quantified blood–brain barrier permeability of intravenously injected 14C-sucrose and 99mTc-albumin. We hypothesized that human blood-derived inter-alpha inhibitor protein treatment would attenuate lipopolysaccharide-induced blood–brain barrier disruption and associated inflammation. Lipopolysaccharide increased blood–brain barrier permeability to both 14C-sucrose and 99mTc-albumin, but human blood-derived inter-alpha inhibitor protein treatment only attenuated increases in 14C-sucrose blood–brain barrier permeability in male mice. Lipopolysaccharide stimulated a more robust elevation of male serum inter-alpha inhibitor protein concentration compared to the elevation measured in female serum. Lipopolysaccharide administration also increased multiple inflammatory factors in serum and brain tissue, including interleukin 6. Human blood-derived inter-alpha inhibitor protein treatment downregulated serum interleukin 6 levels, which were inversely correlated with serum inter-alpha inhibitor protein concentration. We conclude that inter-alpha inhibitor proteins may be neuroprotective through mechanisms of blood–brain barrier disruption associated with systemic inflammation.

Background: IAIPs are serine protease inhibitors that exert anti-inflammatory effects after HI brain injury. We have previously shown that IAIP administration after HI attenuates histopathological brain injury, apoptosis, and microglial activation and improves neurobehavioral outcomes in neonatal rats. Astrocytes are known to form glial scars, increase neuroinflammation, and exacerbate brain injury after stroke. Objective: To determine the effect of IAIPs on astrocytic GFAP expression in the neonatal rat brain after HI. Methods: HI was induced using the Rice-Vannucci method with right carotid ligation and exposure to 8% O 2 for 90 min. Post-natal day 7 rats were assigned to one of three groups: non-ischemic sham-control (Sham, n=12), HI exposed placebo-treated (HI-PL, n=13), or HI exposed IAIP-treated (HI-IAIP, n=14). Intraperitoneal injections of 30 mg/kg of IAIP or saline were given at 15 min, 24, and 48 h after HI. We recorded the sex of the rats. 72 h after HI, brains were collected, paraffin embedded, and sectioned. We performed immunohistochemical analysis by staining slides with GFAP, a marker of astrocyte activation. We performed stereological analysis by capturing images of the whole damaged hemisphere, cerebral cortex, hippocampus, and corpus callosum with the StereoInvestgator 10.0 Probe. We quantified GFAP expression levels using Image J software without knowledge of the group assignments. Results: Sham rats had significantly lower GFAP expression compared with the HI-PL group in the hemisphere (P<0.001), cortex (P<0.005), and hippocampus (P<0.005). IAIP treatment significantly reduced GFAP expression after HI in the hemisphere (P<0.05) compared with the HI-PL group. We also examined GFAP expression separately in male and female rats and found no differences in GFAP expression among the groups in females. However, we found that the Sham males had significantly lower GFAP expression compared to HI-PL in the hemisphere (P<0.005), cortex (P<0.005), and hippocampus (P<0.005), and that the IAIP treated HI exposed males had lower GFAP expression compared with the HI-PL group in the hemisphere (P<0.05) and cortex (P<0.05). Conclusion: IAIPs attenuate HI related astrogliosis in some brain regions particularly in male neonatal rats.