Anti-SARS-CoV-2 Activity of Rhamnan Sulfate from Monostroma nitidum

The COVID-19 pandemic is a major human health concern. The pathogen responsible for COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), invades its host through the interaction of its spike (S) protein with a host cell receptor, angiotensin-converting enzyme 2 (ACE2). In addition to ACE2, heparan sulfate (HS) on the surface of host cells also plays a significant role as a co-receptor. Our previous studies demonstrated that sulfated glycans, such as heparin and fucoidans, show anti-COVID-19 activities. In the current study, rhamnan sulfate (RS), a polysaccharide with a rhamnose backbone from a green seaweed, Monostroma nitidum, was evaluated for binding to the S-protein from SARS-CoV-2 and inhibition of viral infectivity in vitro. The structural characteristics of RS were investigated by determining its monosaccharide composition and performing two-dimensional nuclear magnetic resonance. RS inhibition of the interaction of heparin, a highly sulfated HS, with the SARS-CoV-2 spike protein (from wild type and different mutant variants) was studied using surface plasmon resonance (SPR). In competitive binding studies, the IC₅₀ of RS against the S-protein receptor binding domain (RBD) binding to immobilized heparin was 1.6 ng/mL, which is much lower than the IC₅₀ for heparin (~750 ng/mL). RS showed stronger inhibition than heparin on the S-protein RBD or pseudoviral particles binding to immobilized heparin. Finally, in an in vitro cell-based assay, RS showed strong antiviral activities against wild type SARS-CoV-2 and the delta variant.     

Postfire Succession in Big Sagebrush Steppe With Livestock Grazing

Prescribed fire in rangeland ecosystems is applied for a variety of management objectives, including enhancing productivity of forage species for domestic livestock. In the big sagebrush (Artemisia tridentata Nutt.) steppe of the western United States, fire has been a natural and prescribed disturbance, temporarily shifting vegetation from shrub–grass codominance to grass dominance. There is limited information on the impacts of grazing to community dynamics following fire in big sagebrush steppe. This study evaluated cattle grazing impacts over four growing seasons after prescribed fire on Wyoming big sagebrush (Artemisia tridentata subsp. Wyomingensis [Beetle & Young] Welsh) steppe in eastern Oregon. Treatments included no grazing on burned and unburned sagebrush steppe, two summer-grazing applications after fire, and two spring-grazing applications after fire. Treatment plots were burned in fall 2002. Grazing trials were applied from 2003 to 2005. Vegetation dynamics in the treatments were evaluated by quantifying herbaceous canopy cover, density, annual yield, and perennial grass seed yield. Seed production was greater in the ungrazed burn treatments than in all burn–grazed treatments; however, these differences did not affect community recovery after fire. Other herbaceous response variables (cover, density, composition, and annual yield), bare ground, and soil surface litter did not differ among grazed and ungrazed burn treatments. All burn treatments (grazed and ungrazed) had greater herbaceous cover, herbaceous standing crop, herbaceous annual yield, and grass seed production than the unburned treatment by the second or third year after fire. The results demonstrated that properly applied livestock grazing after low-severity, prescribed fire will not hinder the recovery of herbaceous plant communities in Wyoming big sagebrush steppe.     

Surgical resection of a dysgerminoma in a mare

A mare was referred for further evaluation of a mass found in the left caudal abdomen during a routine postpartum reproductive palpation. The mare was clinically normal with no history of health problems. Ultrasonographic examination of the mass confirmed its presence, but the origin of the mass could not be accurately determined. Routine haematology and serum biochemistry results were within normal limits. The mare was initially treated conservatively with antibiotics, but the mass continued to increase in size, so it was surgically excised. The mass involved the left ovary. The mare showed transient abdominal pain after surgery, but developed no other complications and was in foal 7 months later. On histology, the mass was diagnosed as a dysgerminoma, a rare ovarian tumour of germ cell origin.     

Equine amnionitis and fetal loss: The case definition for an unrecognised cause of abortion in mares

A series of abortions occurred in mares in New South Wales during 2004 that involved similar and unusual findings on post mortem examination of aborted fetuses and fetal membranes. The term Equine Amnionitis and Fetal Loss (EAFL) was developed to describe the condition. This form of abortion had not been previously recognised in Australia. The pathology alone is not specific for EAFL and diagnosis requires demonstration of a combination of certain pathological and bacteriological features. The purpose of this paper is to describe patterns considered consistent with EAFL cases as a working case definition for use by veterinarians and veterinary pathologists in identifying future cases of EAFL. More detailed papers are in preparation to fully describe the epidemiological, histopathological, and microbiological aspects of EAFL.     

Sage Grouse Groceries: Forb Response to Piñon-Juniper Treatments

Juniper and piñon coniferous woodlands have increased 2- to 10-fold in nine ecoregions spanning the Intermountain Region of the western United States. Control of piñon-juniper woodlands by mechanical treatments and prescribed fire are commonly applied to recover sagebrush steppe rangelands. Recently, the Sage Grouse Initiative has made conifer removal a major part of its program to reestablish sagebrush habitat for sage grouse (Centrocercus urophasianus) and other species. We analyzed data sets from previous and ongoing studies across the Great Basin characterizing cover response of perennial and annual forbs that are consumed by sage grouse to mechanical, prescribed fire, and low-disturbance fuel reduction treatments. There were 11 sites in western juniper (Juniperus occidentalis Hook.) woodlands, 3 sites in singleleaf piñon (Pinus monophylla Torr. & Frém.) and Utah juniper (Juniperus osteosperma [Torr.] Little), 2 sites in Utah juniper, and 2 sites in Utah juniper and Colorado piñon (Pinus edulis Engelm). Western juniper sites were located in mountain big sagebrush (A. tridentata ssp. vaseyana) steppe associations, and the other woodlands were located in Wyoming big sagebrush (A. tridentata ssp. wyomingensis) associations. Site potential appears to be a major determinant for increasing perennial forbs consumed by sage grouse following conifer control. The cover response of perennial forbs, whether increasing (1.5- to 6-fold) or exhibiting no change, was similar regardless of conifer treatment. Annual forbs favored by sage grouse benefitted most from prescribed fire treatments with smaller increases following mechanical and fuel reduction treatments. Though forb abundance may not consistently be enhanced, mechanical and fuel reduction conifer treatments remain good preventative measures, especially in phase 1 and 2 woodlands, which, at minimum, maintain forbs on the landscape. In addition, these two conifer control measures, in the short term, are superior to prescribed fire for maintaining the essential habitat characteristics of sagebrush steppe for sage grouse.     

Leukotrienes Affect Secretory Function of Ovarian Cells In Vitro*

The aim of this study was to determine which cells are the source of production and target for leukotriene (LTs) action within the bovine ovary. Luteal (CL, days 14–16 of the oestrous cycle), steroidogenic cells (LSC) and endothelial cells (LEC) of the bovine corpus luteum (CL), and granulosa cells (GC) were isolated enzymatically, cultured in a monolayer and incubated with LTC₄, LTB₄, Azelastine (an antagonist of LTC₄) or Dapsone (an antagonist of LTB₄). Then cells were collected for determination of mRNA expression for LT receptors (LTRs) and 5‐lipoxygenase (5‐LO) by real time RT‐PCR, and media were collected for determination of prostaglandin (PG)E₂, F_2α, progesterone (P4; LSC only), endothelin‐1 (ET‐1; LEC only) and 17‐β oestradiol (E2; GC only). The greatest mRNA expression for LTR‐II and 5‐LO were found in LEC, whereas LTR‐I mRNA expression did not differ among cell types. The level of PGE₂ increased after LTs treatment in each type of ovarian cell, excluding LTC₄ treatment in LEC. The secretion of PGF_2α was also increased by LTs, but decreased after LTB₄ treatment of LSC. In GC cultures, both LTs stimulated E2 secretion; in LEC cultures, LTB₄ stimulated whereas LTC₄ inhibited P4 secretion; in LEC cultures, LTC₄ stimulated but LTB₄ inhibited ET‐1 secretion. The results show that LTs are produced locally and are involved in PGs production/secretion in all examined cells (LSC, LEC and GC) of bovine ovary. Leukotriene treatment modulate secretion of E2, by GC, P4 by LSC and ET‐1 by LEC, which indicates that LTs are involved in regulation of ovarian secretory functions.     

Vegetation Characteristics of Mountain and Wyoming Big Sagebrush Plant Communities in the Northern Great Basin

Dominant plant species are often used as indicators of site potential in forest and rangelands. However, subspecies of dominant vegetation often indicate different site characteristics and, therefore, may be more useful indicators of plant community potential and provide more precise information for management. Big sagebrush (Artemisia tridentata Nutt.) occurs across large expanses of the western United States. Common subspecies of big sagebrush have considerable variation in the types of sites they occupy, but information that quantifies differences in their vegetation characteristics is lacking. Consequently, wildlife and land management guidelines frequently do not differentiate between subspecies of big sagebrush. To quantify vegetation characteristics between two common subspecies of big sagebrush, we sampled 106 intact big sagebrush plant communities. Half of the sampled plant communities were Wyoming big sagebrush (A. tridentata subsp. wyomingensis [Beetle & A. Young] S. L. Welsh) plant communities, and the other half were mountain big sagebrush (A. tridentata subsp. vaseyana [Rydb.] Beetle) plant communities. In general, mountain big sagebrush plant communities were more diverse and had greater vegetation cover, density, and biomass production than Wyoming big sagebrush plant communities. Sagebrush cover was, on average, 2.4-fold higher in mountain big sagebrush plant communities. Perennial forb density and cover were 3.8- and 5.6-fold greater in mountain compared to Wyoming big sagebrush plant communities. Total herbaceous biomass production was approximately twofold greater in mountain than Wyoming big sagebrush plant communities. The results of this study suggest that management guidelines for grazing, wildlife habitat, and other uses should recognize widespread subspecies as indicators of differences in site potentials.     

Expression of the Vascular Endothelial Growth Factor (VEGF‐A) and its Receptors in the Umbilical Cord in the Course of Pregnancy in the Pig

The umbilical cord (UC) and the placenta are important organs through which respiratory gases, nutrients, wastes and biologically active substances are exchanged between the maternal and the foetal system. A rapid placental vascularization observed in the second half of pig pregnancy is positively correlated with the mRNA expression of the vascular endothelial growth factor (VEGF). Based on these findings, we hypothesized that VEGF may have a stimulatory effect in the dynamically growing UC. To further understand the role of the VEGF–VEGFR system during UC development, mRNA and protein expression as well as the cellular localization of VEGF‐A, VEGFR‐1 and VEGFR‐2 in UC were examined on days 40, 60, 75 and 90 of pregnancy and after physiological delivery in the pig (day 114 of pregnancy). Real Time RT‐PCR analysis showed an increase in the mRNA levels of VEGF120 and VEGF164 from day 90 of pregnancy. VEGFR‐1 mRNA expression was significantly increased on day 75 of pregnancy. No significant changes in VEGFR‐2 mRNA expression were detected. In turn, western blot analysis revealed an increase in VEGF‐A protein expression on day 40, compared to the later days of pregnancy. A rapid increase in the VEGFR‐1 protein level was noted on day 75 and 90 of gestation. No significant changes in VEGFR‐2 protein expression were detected on any of the analysed days of pregnancy. Immunohistochemical staining enabled detection of VEGF–VEGFR system, in endothelial and tunica media cells of the umbilical vessels and in allantoic duct and amniotic epithelium on all analysed days of pregnancy. Positive reactions for VEGF‐A and VEGFR‐1, but not VEGFR‐2, were also observed in myofibroblasts. In conclusion, this data shows that members of the VEGF–VEGFR system are temporally and spatially well localized for playing key roles during umbilical cord formation and its intensive growth observed after day 75 of pregnancy.