Water sources and composition of dissolved gases and bubbles in a saline high Arctic spring.

We investigate the water sources for a perennial spring, "Little Black Pond," located at Expedition Fiord, Axel Heiberg Island in the Canadian High Arctic based on dissolved gases. We measured the dissolved O2 in the likely sources Phantom Lake and Astro Lake and the composition of noble gases (3He/4He, 4He, Ne,36Ar, 40Ar, Kr, Xe), N2, O2, CO2, H2S, CH4, and tritium dissolved in the outflow water and bubbles emanating from the spring. The spring is associated with gypsum-anhydrite piercement structures and occurs in a region of thick, continuous permafrost (400-600 m). The water columns in Phantom and Astro lakes are uniform and saturated with O2. The high salinity of the water emanating from the spring, about twice sea water, affects the gas solubility. Oxygen in the water and bubbles is below the detection limit. The N2/Ar ratio in the bubbles and the salty water is 89.9 and 40, respectively, and the relative ratios of the noble gases, with the exception of Neon, are consistent with air dissolved in lake water mixed with air trapped in glacier bubbles as the source of the gases. The Ne/Ar ratio is ~62% of the air value. Our results indicate that about half (0.47±0.1) of the spring water derives from the lakes and the other half from subglacial melt. The tritium and helium results indicate that the groundwater residence time is over 70 years and could be thousands of years.

Mission Architecture Using the SpaceX Starship Vehicle to Enable a Sustained Human Presence on Mars.

A main goal of human space exploration is to develop humanity into a multi-planet species where civilization extends beyond planet Earth. Establishing a self-sustaining human presence on Mars is key to achieving this goal. In situ resource utilization (ISRU) on Mars is a critical component to enabling humans on Mars to both establish long-term outposts and become self-reliant. This article focuses on a mission architecture using the SpaceX Starship as cargo and crew vehicles for the journey to Mars. The first Starships flown to Mars will be uncrewed and will provide unprecedented opportunities to deliver ∼100 metric tons of cargo to the martian surface per mission and conduct robotic precursor work to enable a sustained and self-reliant human presence on Mars. We propose that the highest priority activities for early uncrewed Starships include pre-placement of supplies, developing infrastructure, testing of key technologies, and conducting resource prospecting to map and characterize water ice for future ISRU purposes.

Content of Pb and Zn in Sediments and Hydrobionts as Ecological Markers for Pollution Assessment of Freshwater Objects in Bulgaria-A Review.

The purpose of this review is to describe the contents of Pb and Zn in sediments and hydrobionts as ecological markers for the pollution assessment of freshwater objects in Bulgaria, and the data are compared with other countries and regions. Symmetry was found regarding the levels of Zn in the sediment of the Ovcharitsa and Zhrebchevo dams, which were twice the MAC for arable land (Regulation № 3 of Bulgarian legislation). Symmetry was also observed between the results for Zn and Pb in the studied sediments, and the "favorites" in terms of content were the samples from Zhrebchevo Dam and, especially, from Ovcharitsa Dam. Asymmetry was established in the accumulation of Zn in the livers of carps inhabiting Topolnitsa Dam in comparison with these in Ovcharitsa Dam. A similar asymmetry was observed for lead. The analysis of the muscles and livers of the studied fish showed an asymmetry in the accumulation of zinc, and this process was more intense in the liver. Symmetry was found in the accumulation of Pb in the liver and muscle tissues of the carp from the studied water bodies.

Revision of the species of the genus Diorchis Clerc, 1903 (Cestoda, Hymenolepididae) from rallid birds: a redescription of Diorchis acuminata (Clerc, 1902).

The type-species of the genus Diorchis Clerc, 1903, D. acuminata (Clerc, 1902) Clerc, 1903, is re-described and illustrated on the basis of the type-material from Fulica atra L. from the Middle Ural, Russia, and new specimens from the same host species from Bulgaria. Since the type-series consists of specimens of two species, a syntype is designated as lectotype. The main differentiating characters of D. acuminata are the diorchoid rostellar hooks, 36-39 µm long, with a foliate epiphyseal thickening of the guard; cirrus-sac of variable length, usually reaching and often crossing the midline of proglottis, occasionally reaching antiporal osmoregulatory canals; evaginated cirrus with cylindrical basal part, bulbous middle part and pipette-like distal part; compact vitellarium situated dorsally to the ovary; copulatory part of vagina with muscular poral and middle portions and an antiporal sac-like reservoir; elongate eggs with polar filaments on their envelopes. The type-specimens of D. ransomi Johri, 1939 and D. longibursa Steelman, 1939 from Fulica americana Gmelin from USA are also re-examined and illustrated. Based on the present results, D. ransomi and D. longibursa are recognised as synonyms of D. acuminata. The previous records of the species are discussed. Diorchis acuminata is recognised as a specific parasite of Rallidae (mainly species of the genera Fulica and Gallinula) in the Holarctic.

Protective Mechanism of the Selective Vasopressin V1A Receptor Agonist Selepressin against Endothelial Barrier Dysfunction.

Sepsis and septic shock are among the most common causes of death in the intensive care unit; advanced therapeutic approaches are thus urgently needed. Vascular hyperpermeability represents a major manifestation of severe sepsis and is responsible for the ensuing organ dysfunction and failure. Vasopressin V1A receptor (V1AR) agonists have shown promise in the treatment of sepsis, increasing blood pressure, and reducing vascular hyperpermeability. The effects of the selective V1AR-selective agonist selepressin have been investigated in an in vitro model of thrombin-, vascular endothelial growth factor-, angiopoietin 2-, and lipopolysaccharide (LPS)-induced pulmonary microvascular endothelial hyperpermeability. Results suggest that selepressin counteracts the effects of all four endothelial barrier disruptors in a concentration-dependent manner, as reflected in real-time measurements of vascular permeability by means of transendothelial electrical resistance. Further, selepressin protected the barrier integrity against the LPS-mediated corruption of the endothelial monolayer integrity, as captured by VE-cadherin and actin staining. The protective effects of selepressin were abolished by silencing of the vasopressin V1AR, as well as by atosiban, an antagonist of the human V1AR. p53 appears to be involved in mediating these palliative effects, since selepressin strongly induced its expression levels, suppressed the inflammatory RhoA/myosin light chain2 pathway, and triggered the barrier-protective effects of the GTPase Rac1. We conclude that V1AR-selective agonists, such as selepressin, may prove useful in the improvement of endothelial barrier function in the management of severe sepsis. SIGNIFICANCE STATEMENT: A cardinal sign of sepsis, a serious disease with significant mortality and no specific treatment, is pulmonary endothelial barrier dysfunction that leads to pulmonary edema. Here, we present evidence that in cultured human lung microvascular endothelial cells, the synthetic, selective vasopressin V1A receptor agonist selepressin protects against endothelial barrier dysfunction caused by four different edemogenic agents, suggesting a potential role of selepressin in the clinical management of sepsis.

The HSP90 Inhibitor, AUY-922, Ameliorates the Development of Nitrogen Mustard-Induced Pulmonary Fibrosis and Lung Dysfunction in Mice.

Increased levels of heat shock protein 90 (HSP90) have been recently implicated in the pathogenesis of pulmonary fibrosis and the use of HSP90 inhibitors constitutes a potential therapeutic approach. Similarly, acute exposure to nitrogen mustard (NM) is related to the development of chronic lung injury driven by TNF-α, TGF-ß, ERK and HSP90. Thus, we developed a murine model of NM-induced pulmonary fibrosis by instilling C57BI/6J mice with 0.625 mg/kg mechlorethamine hydrochloride. After 24 h, mice began receiving AUY-922, a second generation HSP90 inhibitor, at 1 mg/kg 2 times per week or 2 mg/kg 3 times per week, for either 10 or 30 days. AUY-922 suppressed the NM-induced sustained inflammation, as reflected in the reduction of leukocyte and protein concentrations in bronchoalveolar lavage fluid (BALF), and inhibited the activation of pro-fibrotic biomarkers, ERK and HSP90. Furthermore, AUY-922 maintained normal lung function, decreased the overexpression and accumulation of extracellular matrix proteins, and dramatically reduced histologic evidence of fibrosis in the lungs of mice exposed to NM. The HSP90 inhibitor, AUY-922, successfully blocked the adverse effects associated with acute exposures to NM, representing a promising approach against NM-induced pulmonary fibrosis.

Post-treatment with a heat shock protein 90 inhibitor prevents chronic lung injury and pulmonary fibrosis, following acute exposure of mice to HCl.

Aim/Purpose: Exposure to high levels of hydrochloric acid (HCl) is associated with severe lung injury including both acute inflammation and chronic lung disease, which leads to the development of pulmonary fibrosis. Currently, there are no specific therapeutic agents for HCl-induced lung injury. Heat shock protein 90 (HSP90) has been implicated in the pathogenesis of pulmonary fibrosis. Thus, we have used a murine model of intra-tracheal acid instillation to investigate the antidotal effects of AUY-922, a small molecule HSP90 inhibitor, already in clinical trials for various types of cancer, against HCl-induced chronic lung injury and pulmonary fibrosis.Methods: HCl (0.1 N, 2 µl/g body weight) was instilled into male C57Bl/6J mice at day 0. After 24 h, mice began receiving 1 mg/kg AUY-922, 2x/week for 15 or 30 days.Results: AUY-922 suppressed the HCl-induced sustained inflammation, as reflected in the reduction of leukocyte and protein concentrations in bronchoalveolar lavage fluid, and inhibited the activation of pro-fibrotic biomarkers, ERK and HSP90. Furthermore, AUY-922 improved lung function, decreased the overexpression and accumulation of extracellular matrix proteins and dramatically reduced histologic evidence of fibrosis in the lungs of mice exposed to HCl.Conclusions: We conclude that AUY-922, and possibly other HSP90 inhibitors, successfully block the adverse effects associated with acute exposures to HCl and may represent an effective antidote against HCl-induced chronic lung injury and fibrosis.

Development of chronic lung injury and pulmonary fibrosis in mice following acute exposure to nitrogen mustard.

Objective: Sulfur mustards are toxic agents used as a chemical warfare in the twentieth century. Exposure to nitrogen mustards (NM), their more water-soluble analogs, is associated with respiratory, dermatological, neurological, and systemic symptoms whose severity depends on dose and length of contact. Long-term effects of acute inhalation have been related to the development of chronic lung injury and pulmonary fibrosis whose precise mechanisms and potential antidotes are yet to be discovered.Materials and methods: We have developed a model of NM-induced pulmonary fibrosis by intratracheally instilling mechlorethamine hydrochloride into C57Bl/6J male mice.Results and Discussion: Following mechlorethamine exposure, strong early and milder late inflammatory responses were observed. Initially, the number of white blood cells and levels of protein and pro-inflammatory cytokines in the bronchoalveolar lavage fluid (BALF) increased, followed by increases in the number of macrophages and the levels of transforming growth factor-ß (TGF-ß), a pro-fibrotic mediator. Analysis of lung homogenates revealed increased phosphorylation of pro-fibrotic biomarkers, serine/threonine-selective protein kinases (p-ERK), and heat shock protein 90 (P-HSP90) at 10 and 30 days after exposure. Total collagen expression and deposition of extracellular matrix proteins also increased. Lung function measurements demonstrated the presence of both obstructive and restrictive disease in agreement with evidence of increased lower airway peribronchial collagen deposition and parenchymal fibrosis.Conclusions: We conclude that the mouse represents a useful model of NM-induced acute lung injury and chronic pulmonary fibrosis, the latter driven by the overexpression of TGF-ß, p-ERK, and P-HSP90. This model may prove useful in the pre-clinical development of antidotes and other countermeasures.

Acute exposure of mice to hydrochloric acid leads to the development of chronic lung injury and pulmonary fibrosis.

Objective: Accidental exposure to hydrochloric acid (HCl) is associated with acute lung injury in humans, development of long-term chronic airway obstruction, and fibrosis. However, the mechanisms responsible for the progression to pulmonary fibrosis remain unclear. We utilized a mouse model of progressive lung injury from a single exposure to HCl to investigate the effects of HCl on the lower respiratory tract. Materials and methods: HCl (0.05-0.3 N) or saline was injected intratracheally into male C57Bl/6J mice. At 1, 4, 10 and 30 days post instillation, bronchoalveolar lavage fluid (BALF) and lung tissues were collected and examined for multiple outcomes. Results and discussion: We observed an early inflammatory response and a late mild inflammation present even at 30 d post HCl exposure. Mice treated with HCl exhibited higher total leukocyte and protein levels in the BALF compared to the vehicle group. This was characterized by increased number of neutrophils, monocytes, and lymphocytes as well as pro-inflammatory cytokines during the first 4 d of injury. The late inflammatory response exhibited a predominant presence of mononuclear cells, increased permeability to protein, and higher levels of the pro-fibrotic mediator TGFß. Pro-fibrotic protein biomarkers, phosphorylated ERK, and HSP90, were also overexpressed at 10 and 30 d following HCl exposure. In vivo lung function measurements demonstrated lung dysfunction and chronic lung injury associated with increased lung hydroxyproline content and increased expression of extracellular matrix (ECM) proteins. The acute inflammation and severity of fibrosis increased in HCl-concentration dependent manner. Conclusions: Our findings suggest that the initial inflammatory response and pro-fibrotic biomarker upregulation may be linked to the progression of pulmonary fibrosis and airway dysfunction and may represent valuable therapeutic targets.

Comparison of Purified ß-glucuronidases in Patient Urine Samples Indicates a Lack of Correlation Between Enzyme Activity and Drugs of Abuse Metabolite Hydrolysis Efficiencies Leading to Potential False Negatives.

Pain management laboratories analyze biological fluids (urine, saliva or blood) from patients treated for chronic pain to ensure compliance and to detect undisclosed drug use. The quantitation of multi-panel drugs in urine and tissues utilizes ß-glucuronidase to cleave the glucuronic acid and liberate the parent drug for mass spectrometry analysis. This work focuses on the comparison of three different, purified and commercially available ß-glucuronidases across 83 patient urine samples. One enzyme is genetically modified, expressed in bacteria and the other two enzymes are purified from abalone. The results indicate that the source of ß-glucuronidase plays an important role in substrate specificity which in turn dictates hydrolysis efficiency. Contaminants in the enzyme solutions also interfere with analyte detection. Altogether, these factors impact precision and accuracy of data interpretation, leading up to 13% positive/negative disagreement.

Increased activity of ß-glucuronidase variants produced by site-directed mutagenesis.

ß-glucuronidase (BGus) is an essential glycosyl hydrolase which has been widely used in biological and biomedical applications. In this paper, we report the construction and screening of nineteen Escherichia coli BGus (EBGus) mutants using site-directed mutagenesis. The mutants G559N, G559S and G559T showed a 3-5 fold increase in enzyme activity in comparison to wild type EBGus. In particular, G559S, with the highest activity, showed 2-6 fold enhanced activity compared to abalone and snail BGus extracts. Moreover, the glycine to serine mutagenesis for the same site in Staphylococcus sp. RLH1 BGus (StBGus) exhibited significantly enhanced activity, which indicated the importance of the G559→S mutation on BGus function. Based on this structural analysis, we postulate that the mutation at G559 plays an important role in the stabilization of the enzyme conformation, and thereby facilitates the effective binding of substrate.

Degradation of Opioids and Opiates During Acid Hydrolysis Leads to Reduced Recovery Compared to Enzymatic Hydrolysis.

Drug monitoring laboratories utilize a hydrolysis process to liberate the opiates from their glucuronide conjugates to facilitate their detection by tandem mass spectrometry (MS). Both acid and enzyme hydrolysis have been reported as viable methods, with the former as a more effective process for recovering codeine-6-glucuronide and morphine-6-glucuronide. Here, we report concerns with acid-catalyzed hydrolysis of opioids, including a significant loss of analytes and conversions of oxycodone to oxymorphone, hydrocodone to hydromorphone and codeine to morphine. The acid-catalyzed reaction was monitored in neat water and patient urine samples by liquid chromatography-time-of-flight and tandem MS. These side reactions with acid hydrolysis may limit accurate quantitation due to loss of analytes, possibly lead to false positives, and poorly correlate with pharmacogenetic profiles, as cytochrome P450 enzyme (CYP2D6) is often involved with oxycodone to oxymorphone, hydrocodone to hydromorphone and codeine to morphine conversions. Enzymatic hydrolysis process using the purified, genetically engineered ß-glucuronidase (IMCSzyme®) addresses many of these concerns and demonstrates accurate quantitation and high recoveries for oxycodone, hydrocodone, oxymorphone and hydromorphone.

Nearing the cold-arid limits of microbial life in permafrost of an upper dry valley, Antarctica.

Some of the coldest and driest permafrost soils on Earth are located in the high-elevation McMurdo Dry Valleys (MDVs) of Antarctica, but little is known about the permafrost microbial communities other than that microorganisms are present in these valleys. Here, we describe the microbiology and habitable conditions of highly unique dry and ice-cemented permafrost in University Valley, one of the coldest and driest regions in the MDVs (1700 m above sea level; mean temperature -23 °C; no degree days above freezing), where the ice in permafrost originates from vapour deposition rather than liquid water. We found that culturable and total microbial biomass in University Valley was extremely low, and microbial activity under ambient conditions was undetectable. Our results contrast with reports from the lower-elevation Dry Valleys and Arctic permafrost soils where active microbial populations are found, suggesting that the combination of severe cold, aridity, oligotrophy of University Valley permafrost soils severely limit microbial activity and survival.

Description of Diorchis thracica n. sp. (Cestoda, Hymenolepididae) from the ruddy shelduck Tadorna ferruginea (Pallas) (Anseriformes, Anatidae) in Bulgaria.

Diorchis thracica n. sp. (Cestoda, Cyclophyllidea, Hymenolepididae) is described from the ruddy shelduck Tadorna ferruginea (Pallas) (Aves, Anseriformes, Anatidae), collected in the vicinities of Radnevo, Stara Zagora Region, Bulgaria. The new species is differentiated from other members of Diorchis Clerc, 1903 by possessing rostellar hooks with length of 36 µm, a thick-walled cirrus-sac with strong longitudinal muscular fibres in its middle part and a copulatory vagina with two sphincters. Main morphological criteria for distinguishing species of the genus Diorchis are discussed.

The Icebreaker Life Mission to Mars: a search for biomolecular evidence for life.

The search for evidence of life on Mars is the primary motivation for the exploration of that planet. The results from previous missions, and the Phoenix mission in particular, indicate that the ice-cemented ground in the north polar plains is likely to be the most recently habitable place that is currently known on Mars. The near-surface ice likely provided adequate water activity during periods of high obliquity, ≈ 5 Myr ago. Carbon dioxide and nitrogen are present in the atmosphere, and nitrates may be present in the soil. Perchlorate in the soil together with iron in basaltic rock provides a possible energy source for life. Furthermore, the presence of organics must once again be considered, as the results of the Viking GCMS are now suspect given the discovery of the thermally reactive perchlorate. Ground ice may provide a way to preserve organic molecules for extended periods of time, especially organic biomarkers. The Mars Icebreaker Life mission focuses on the following science goals: (1) Search for specific biomolecules that would be conclusive evidence of life. (2) Perform a general search for organic molecules in the ground ice. (3) Determine the processes of ground ice formation and the role of liquid water. (4) Understand the mechanical properties of the martian polar ice-cemented soil. (5) Assess the recent habitability of the environment with respect to required elements to support life, energy sources, and possible toxic elements. (6) Compare the elemental composition of the northern plains with midlatitude sites. The Icebreaker Life payload has been designed around the Phoenix spacecraft and is targeted to a site near the Phoenix landing site. However, the Icebreaker payload could be supported on other Mars landing systems. Preliminary studies of the SpaceX Dragon lander show that it could support the Icebreaker payload for a landing either at the Phoenix site or at midlatitudes. Duplicate samples could be cached as a target for possible return by a Mars Sample Return mission. If the samples were shown to contain organic biomarkers, interest in returning them to Earth would be high.

Debittering of Protein Hydrolysates by Lactobacillus LBL-4 Aminopeptidase.

Yoghurt strain Lactobacillus LBL-4 cultivated for 8-10 h at pH ~6.0 was investigated as a considerable food-grade source of intracellular aminopeptidase. Cell-free extract manifesting >200 AP U/l was obtained from cells harvested from 1 L culture media. Subtilisin-induced hydrolysates of casein, soybean isolate, and Scenedesmus cell protein with degree of hydrolysis 20-22% incubated at 45°C for 10 h by 10 AP U/g peptides caused an enlarging of DH up to 40-42%, 46-48%, and 38-40% respectively. The DH increased rapidly during the first 4 h, but gel chromatography studies on BioGel P-2 showed significant changes occurred during 4-10 h of enzyme action when the DH increased gradually. After the digestion, the remained AP activity can be recovered by ultrafiltration (yield 40-50%). Scenedesmus protein hydrolysate with DH 20% was inoculated by Lactobacillus LBL-4 cells, and after 72 h cultivation the DH reached 32%. The protein hydrolysates (DH above 40%) obtained from casein and soybean isolate (high Q value) demonstrated a negligible bitterness while Scenedesmus protein hydrolysates (low Q value) after both treatments were free of bitterness.

Preparation of antioxidant enzymatic hydrolysates from honeybee-collected pollen using plant enzymes.

Enzymatic hydrolysates of honeybee-collected pollen were prepared using food-grade proteinase and aminopeptidases entirely of plant origin. Bromelain from pineapple stem was applied (8 mAU/g substrate) in the first hydrolysis stage. Aminopeptidase (0.05 U/g substrate) and proline iminopeptidase (0.03 U/g substrate) from cabbage leaves (Brassica oleracea var. capitata), and aminopeptidase (0.2 U/g substrate) from chick-pea cotyledons (Cicer arietinum L.) were involved in the additional hydrolysis of the peptide mixtures. The degree of hydrolysis (DH), total phenolic contents, and protein contents of these hydrolysates were as follows: DH (about 20-28%), total phenolics (15.3-27.2 µg/mg sample powder), and proteins (162.7-242.8 µg/mg sample powder), respectively. The hydrolysates possessed high antiradical scavenging activity determined with DPPH (42-46% inhibition). The prepared hydrolysates of bee-collected flower pollen may be regarded as effective natural and functional dietary food supplements due to their remarkable content of polyphenol substances and significant radical-scavenging capacity with special regard to their nutritional-physiological implications.

Heavy metals in Antarctic notothenioid fish from South Bay, Livingston Island, South Shetlands (Antarctica).

The Pb, Cd, Cu, Zn, and Mn contents of the liver, spleen, muscle, bones, scales, gills, and the whole body of 3- to 7-year-old notothenioid Antarctic cod (Notothenia coriiceps, Richardson, 1844) were measured. The highest heavy metal concentrations obtained are as follows: Cd in liver, the mean value was 1.36 ± 0.19 mg/kg dry weight (wt); Pb and Zn in spleen, the mean values were 3.33 ± 0.86 and 143.97 ± 16.17 mg/kg dry wt, respectively; Cu in gills, 3.76 ± 1.16 mg/kg dry wt; and Mn in scales, 14.80 ± 4.77 mg/kg dry wt. The comparison with the data reported up to now shows that the metal concentrations varied within relative wide ranges. These first data obtained could be used as a baseline to investigate further relationships among metal contents in fish, their diet, and habitat.

Purification and characterization of L-phenylalanine aminopeptidase from chick-pea cotyledons (Cicer arietinum L.).

Chick-pea (Cicer arietinum L.) cotyledons are unique source of aminopeptidase - 8-9 U/g cotyledons was observed using L-leucine-p-nitroanilide as substrate. The aminopeptidase was purified (65 kDa, pI 4.8 ) reaching a specific activity of 220 U/mg at pH 7.0-7.2 and 35-40 degrees C. The determined constant of specificity k(cat)/K(m) during hydrolysis of N-unsubstituted amino acid-p-nitroanilides showed a decrease order: Phe>Leu>Pro>Ile>Val>Ala. The enzyme was strongly inhibited by p-chloromercuribenzoic acid as well as in a competitive rate by the antihypertensive peptides Ile-Pro-Pro and Val-Pro-Pro.

Mega-impact formation of the Mars hemispheric dichotomy.

The Mars hemispheric dichotomy is expressed as a dramatic difference in elevation, crustal thickness and crater density between the southern highlands and northern lowlands (which cover approximately 42% of the surface). Despite the prominence of the dichotomy, its origin has remained enigmatic and models for its formation largely untested. Endogenic degree-1 convection models with north-south asymmetry are incomplete in that they are restricted to simulating only mantle dynamics and they neglect crustal evolution, whereas exogenic multiple impact events are statistically unlikely to concentrate in one hemisphere. A single mega-impact of the requisite size has not previously been modelled. However, it has been hypothesized that such an event could obliterate the evidence of its occurrence by completely covering the surface with melt or catastrophically disrupting the planet. Here we present a set of single-impact initial conditions by which a large impactor can produce features consistent with the observed dichotomy's crustal structure and persistence. Using three-dimensional hydrodynamic simulations, large variations are predicted in post-impact states depending on impact energy, velocity and, importantly, impact angle, with trends more pronounced or unseen in commonly studied smaller impacts. For impact energies of approximately (3-6) x 10(29) J, at low impact velocities (6-10 km s(-1)) and oblique impact angles (30-60 degrees ), the resulting crustal removal boundary is similar in size and ellipticity to the observed characteristics of the lowlands basin. Under these conditions, the melt distribution is largely contained within the area of impact and thus does not erase the evidence of the impact's occurrence. The antiquity of the dichotomy is consistent with the contemporaneous presence of impactors of diameter 1,600-2,700 km in Mars-crossing orbits, and the impact angle is consistent with the expected distribution.