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Various issues concerned with the assessment of substance use and abuse phenotypes are outlined.

• INRA96 extender supplemented with liposomes of egg yolk phospholipids improves significantly motility parameters Among biotechnologies of reproduction in the equine species, artificial insemination remains the most used technology especially for cooled transported sperm. Although the use of INRA96 extender has demonstrated its efficiency for long-term sperm storage at 4°C or 15°C, some stallions ("bad coolers") are excluded from such technology. Some years ago, we demonstrated that liposomes produced from egg yolk (EY) phospholipids could be an alternative to egg yolk plasma in stallion freezing extenders. To develop a new extender for sperm chilling, we evaluated the protective effect of liposomes produced from EY phospholipids on stallion sperm storage at 4°C. The sperm of stallions from two studs was diluted in INRA96 extender (as control) or an experimental extender (EE) composed of INRA96 supplemented with liposomes of EY phospholipids. After 24H (D1), 72H (D3), and 6 days (D6) or 7 days (D7), motility parameters were evaluated using Computer Assisted Semen Analyzer. Our results demonstrated that total and progressive motility decreased significantly after dilution and storage in INRA96 between D1 and D3 (P <.05) while no significant decrease was observed between D1 and D3 with EE. Regarding VAP parameter, no significant difference was observed between extenders except at D7 in stud 2. Moreover, total and progressive motility were maintained at a significantly higher level (D3, D6, D7) when sperm was stored in EE compared to INRA96. These promising results demonstrate that the supplementation of INRA96 extender with egg-yolk phospholipids liposomes allows a higher protection to stallion sperm cells. [ABSTRACT FROM AUTHOR]

Background: Political rhetoric from the former U.S. president influences Latinx adults' feelings of safety and their decisions to seek care in the emergency department (ED).Objective: Our aim was to examine the impact of political rhetoric on feelings of safety and health care access in the pediatric population.Methods: This was a cross-sectional study of undocumented Latinx families (ULF), Latinx U.S. citizen families (LCF), and non-Latinx U.S. citizen families (NLF) conducted from November 2018 through February 2020 by means of interviewing a convenience sample of parents and guardians who brought their child to a pediatric clinic and two EDs in California.Results: Of 705 parents approached, 449 (63.7%) agreed to participate: 138 ULF, 150 LCF, and 158 NLF. Most ULF (95%), LCF (88%), and NLF (78%) parents and guardians had heard anti-immigrant statements from the former U.S. president and most (94% ULF, 90% LCF, 86% NLF) believed these measures against immigrants were being enacted or will be enacted. More ULF (75%, 95% confidence interval [CI] 67-81%) reported that these statements made them concerned about their child's safety in the United States compared with 36% (95% CI 28-45%) and 34% (95% CI 26-43%) of LCF and NLF, respectively. More ULF 17% (95% CI 11-24%) said that these statements made them afraid to bring their child for medical care, compared with 5% (95% CI 2-10%) and 3% (95% CI 1-7%) of LCF and NLF, respectively.Conclusions: Most parents heard statements against undocumented immigrants by the former U.S. president and most believed measures were being enacted. This rhetoric had a substantial negative impact on ULF parents in terms of safety concerns for their child and fear of accessing health care. [ABSTRACT FROM AUTHOR]

• First study to mimic black soldier fly larva (BSFL) midgut digestion in vitro. • Digestion products can indicate biowaste treatment performance. • In vitro digestions can be a screening tool for treatment design. • First study to determine BSFL diet intake and midgut region residence times. • Biowaste protein and carbohydrates influence midgut residence time. Black soldier fly larvae treatment is an emerging technology for the conversion of biowaste into potentially more sustainable and marketable high-value products, according to circular economy principles. Unknown or variable performance for different biowastes is currently one challenge that prohibits the global technology up-scaling. This study describes simulated midgut digestion for black soldier fly larvae to estimate biowaste conversion performance. Before simulation, the unknown biowaste residence time in the three midgut regions was determined on three diets varying in protein and non-fiber carbohydrate content. For the static in vitro model, diet residence times of 15 min, 45 min, and 90 min were used for the anterior, middle, and posterior midgut region, respectively. The model was validated by comparing the ranking of diets based on in vitro digestion products to the ranking found in in vivo feeding experiments. Four artificial diets and five biowastes were digested using the model, and diet digestibility and supernatant nutrient contents were determined. This approach was able to distinguish broadly the worst and best performing rearing diets. However, for some of the diets, the performance estimated based on in vitro results did not match with the results of the feeding experiments. Future studies should try to establish a stronger correlation by considering fly larvae nutrient requirements, hemicellulose digestion, and the diet/gut microbiota. In vitro digestion models could be a powerful tool for academia and industry to increase conversion performance of biowastes with black soldier fly larvae. [ABSTRACT FROM AUTHOR]

Assisted reproductive technologies (ART) are widely used for both humans and domestic animals. In bovine species, in vitro embryo production is increasingly used and significant efforts are being made to optimize media and culture conditions. Phospholipase A2 (PLA2) are lipolytic enzymes that hydrolyze glycerophospholipids to produce free fatty acids and lysophospholipids that have been found to be critical for many biological processes. Mouse group X secreted PLA 2 (mGX) is abundant in the male reproductive tract and its use during sperm capacitation has been shown to improve in vitro production of viable embryos in a mouse model. Here, we examined its effect in the bovine species, testing the impact of mGX on the three steps involved in vitro production of preimplantation embryos: oocyte maturation, fertilization and preimplantation development. We found that incubating cumulus oocyte complexes (COC) or gametes with mGX resulted in increased blastocyst hatching and blastocyst production, respectively. The increases of embryo production induced by the phospholipase mGX were not observed for the catalytically inactive mutant H48Q-mGX, suggesting that these effects require the enzymatic activity of mGX. We also tested bGIB, a bovine homolog of mGX. bGIB failed to improve blastocyst production, underlining the high specificity of mGX. In conclusion, the results presented show that the effects of mGX are not restricted to the mouse model and that it is potent in the bovine species as well. This result strengthens the potential of mGX as a "pro-fertility drug" for mammalian reproduction. • secreted group X PLA2 applied during oocyte in vitro maturation improves embryo development. • secreted group X PLA2 applied during in vitro fertilization improves embryo development. • Effects of group X sPLA2 include increase of blastocyst formation, expansion and hatching. • Effects of group X sPLA2 is not due to LPA production. • Group IB sPLA2 does not mimic effects of group X sPLA2. [ABSTRACT FROM AUTHOR]

Abstract: Twenty semen samples taken from 5 dogs were frozen in liquid nitrogen at −196°C in four different extenders: one control extender based on 20% egg yolk, 6% LDL alone (low density lipoproteins: the active cryoprotective principle in chicken egg yolk), 6% LDL combined with 20mmol glutamine, and Equex® (a reference extender that we wish to compare with the LDL-glutamine combination). After thawing, spermatozoal motility was evaluated using a HAMILTON THORNE CERROS 12 image analyzer; the percentage of motile spermatozoa was 27.7% in the egg yolk extender (p <0.05), 49.9% with 6% LDL alone (p >0.05), 54.7% in the 6% LDL+20mmol glutamine extender, and 47.9% with Equex® (p >0.05). The motility parameters (VAP, VCL, VSL and ALH) were also superior in the 6% LDL+20mmol glutamine extender in comparison with the other extenders. Finally, the spermatozoa were generally better protected during freezing with the 6% LDL+20mmol glutamine association than with the egg yolk, 6% LDL, or Equex extenders in terms of the flagellar plasma membrane (HOS test), DNA (Acridine orange test), and acrosome integrity (Spermac® test: no significant difference). The Equex® extender obtained the best results for the acrosome, followed by 6% LDL+20mmol glutamine (FITC-PSA test: p <0.05 between each extender). [Copyright &y& Elsevier]

Abstract: Egg yolk is normally used as a protective agent to freeze semen of equine and other species. However, addition of egg yolk in extenders is not without disadvantages and the demand to find cryoprotective alternatives is strong. The objective of this study was to test the cryoprotective capacities of liposomes composed of egg yolk phospholipids. Two experiments were conducted: 1) the first to determine the optimal composition and concentration of liposomes to preserve post-thaw motility and membrane integrity of spermatozoa; 2) the second to assess in vivo the cryoprotective capacities of these liposomes. In Experiment 2, post-thaw motility and membrane integrity of spermatozoa were also analyzed. Experiment 1 demonstrated that liposomes composed of phospholipids E80 (commercial lecithins from egg yolk composed mainly of phosphatidylcholine and phosphatidylethanolamine) and of Hank''s salts-glucose-lactose solution (E80-liposomes) were the most efficient in preserving post-thaw motility. The optimal concentration was 4 % (v/v). In Experiment 2, fertility rate after artificial insemination of semen frozen with E80-liposomes was 55 % (22/40) compared with 68 % (27/40) with the control extender containing egg yolk (EY) (p = 0.23). Post-thaw motility parameters were higher with EY than with E80-liposomes (p < 0.0001). For post-thaw membrane integrity no difference was observed between the two extenders (p = 0.08). Liposomes composed of egg yolk phospholipids appeared to be a promising alternative to replace egg yolk in semen freezing extenders in equine species. [Copyright &y& Elsevier]

Abstract: Chicken egg yolk is held as an excellent cryoprotective agent for freezing canine semen. Recent advances have enabled the extraction of low density lipoproteins from egg yolk, which are responsible for the cryoprotective abilities of the latter. The objective of this article was to compare 3 semen extenders for freezing canine semen: 2 containing egg yolk (Tris egg yolk and Equex STAMP) and one containing 6% LDL. After freezing and thawing 20 ejaculates from 5 different dogs, the 6% LDL extender produced 50% mobile spermatozoa, compared with 48% with the Equex® extender and 27.7% with the extender containing egg yolk alone (EY). In vitro functional tests demonstrated that the integrity of the plasma membrane (hypoosmotic test) was respected in 65–66% of spermatozoa as a function of the extender; DNA integrity was respected in more than 97% of the spermatozoa. The Equex® extender provided superior acrosome integrity (FITC/PSA test): 68.4% compared with 55.1% with LDL and 53.3% with egg yolk. However, the 6% LDL extender resulted in fewer spermatozoal anomalies (Spermac® test), with 54.6% normal spermatozoa compared to 53.6% for Equex® and 53.3% with the egg yolk. All six of the bitches inseminated artificially via the intra-uterine route (Scandinavian technique) using semen frozen in the 6% LDL extender became pregnant. The LDL extender resulted in percentages of mobile spermatozoa and movement characteristics that were as good if not better than those obtained with the reference extenders following thawing. The 6% LDL extender appears to have the same cryoprotective qualities as the reference diluent, Equex® STAMP. [Copyright &y& Elsevier]

Abstract: Deoxyguanosine kinase (DGUOK) catalyzes the first step of the mitochondrial deoxypurine salvage pathway, the phosphorylation of purine deoxyribonucleosides. Mutations in the DGUOK gene have been linked to inherited mtDNA depletion syndromes, neonatal liver failure, nystagmus, and hypotonia. Previously, we reported the first case of a heterozygous unclassified c.592-4_c.592-3delTT alteration in a patient with DGUOK deficiency without the demonstration of its pathogenicity (). This alteration was predicted to cause aberrant splicing based upon two computer algorithms. We now report a homozygous c.592-4_c.592-3delTT mutation found in two affected siblings of asymptomatic consanguineous parents. The proband presented with symptoms of idiopathic hepatitis, liver dysfunction, nystagmus, and retinal blindness. This individual died at 6months of age due to liver failure. This individual’s affected sibling presented similarly and has remarkable elevations of tyrosine, methionine, and alanine. Many organic acids were elevated in urine, including lactic acid, Krebs cycle intermediates, and para-hydroxy compounds; ketone bodies were also present. RNA studies support aberrant splicing. Sequencing of cDNA detected exon 5 skipping in the two affected siblings, but not in the normal control. These results indicate that the homozygous c.592-4_c.592-3delTT is deleterious and responsible for the DGUOK deficiency. The parents were subsequently confirmed to be carriers of this mutation. In summary, we have demonstrated that c.592-4_c.592-3delTT is a pathogenic splice acceptor site mutation leading to DGUOK deficiency. [Copyright &y& Elsevier]

The purpose of this work was to investigate the contribution of thermodynamics and mobility to the physical stability of two pharmaceutical glasses with similar glass transition temperatures (Tg), by comparing configurational thermodynamic quantities and molecular relaxation time constants (τ) at temperatures below Tg. Ritonavir and nifedipine were chosen as model glasses because they show excellent and poor physical stability, respectively. Although ritonavir and nifedipine have similar Tg values (50 and 46°C, respectively), amorphous ritonavir is quite stable while nifedipine has been reported to crystallize at temperatures as low as 40°C below Tg. Modulated temperature differential scanning calorimetry (MTDSC) was used to characterize both crystalline phases and freshly prepared glasses. The glasses were then annealed at Tg-Ta = 25°C while monitoring the extent of relaxation and heat capacity change as a function of time via MTDSC. Configurational thermodynamic quantities (Gc, Hc, and Sc) and molecular relaxation time constants, τ, were calculated from the calorimetric data. Interestingly, the Gibbs free energy driving force for crystallization was nearly identical for the two compounds. The largest differences were found in the configurational entropy (Sc) values for the fresh glasses and in the Sc values over time. Configurational entropy values were approximately 50% higher for ritonavir. The τ values of freshly prepared glasses indicated that both materials had similar initial mobility at the annealing temperatures and the temperature dependence of τ was approximately Arrhenius, regardless of age. Although initial τ values were similar, the τ values after 3 days annealing were ∼sixfold greater for ritonavir. The relatively poor physical stability of nifedipine compared to ritonavir is attributed to both the lower entropic barrier to crystallization for fresh and annealed glass, and higher molecular mobility in aged glasses of nifedipine. These observations below Tg are consistent with the previous work on physical stability of amorphous pharmaceuticals performed above Tg. © 2006 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 96:71–83, 2007 [ABSTRACT FROM AUTHOR]

Most plant-pollinator networks are based on observations of contact between an insect and a flower in the field. Despite significant sampling efforts, some links are easier to report, while others remain unobserved. Therefore, visit-based networks represent a subsample of possible interactions in which the ignored part is variable. Pollen is a natural marker of insect visits to flowers. The identification of pollen found on insect bodies can be used as an alternative method to study plant-pollinator interactions, with a potentially lower risk of bias than the observation of visits, since it increases the number of interactions in the network. Here we compare plant-pollinator networks constructed (i) from direct observation of pollinator visits and (ii) from identification of pollen found on the same insects. We focused on three calcareous grasslands in France, with different plant and pollinator species diversities. Since pollen identification always yields richer, more connected networks, we focused our comparisons on sampling bias at equal network connectance. To do so, we first compared network structures with an analysis of latent blocks and motifs. We then compared species roles between both types of networks with an analysis of specialization and species positions within motifs. Our results suggest that the sampling from observations of insect visits does not lead to the construction of a network intrinsically different from the one obtained using pollen found on insect bodies, at least when field sampling strives to be exhaustive. Most of the significant differences are found at the species level, not at the network structure level, with singleton species accounting for a respectable fraction of these differences. Overall, this suggests that recording plant-pollinator interactions from pollinator visit observation does not provide a biased picture of the network structure, regardless of species richness; however, it provided less information on species roles than the pollen-based network. • Visit-based networks were compared to simulated pollen-based networks in three sites. • Network structures do not differ significantly between the two types of networks. • Some species roles and specialization changed significantly with pollen information. • Overall, observation of visits does not provide a biased picture of plant-pollinator networks. [ABSTRACT FROM AUTHOR]