COVID-19 and vertical transmission: assessing the expression of ACE2/TMPRSS2 in the human fetus and placenta to assess the risk of SARS-CoV-2 infection.

BACKGROUND: Pregnant women have been identified as a potentially at-risk group concerning COVID-19 infection, but little is known regarding the susceptibility of the fetus to infection. Co-expression of ACE2 and TMPRSS2 has been identified as a prerequisite for infection, and expression across different tissues is known to vary between children and adults. However, the expression of these proteins in the fetus is unknown. METHODS: We performed a retrospective analysis of a single cell data repository. The data were then validated at both gene and protein level by performing RT-qPCR and two-colour immunohistochemistry on a library of second-trimester human fetal tissues. FINDINGS: TMPRSS2 is present at both gene and protein level in the predominantly epithelial fetal tissues analysed. ACE2 is present at significant levels only in the fetal intestine and kidney, and is not expressed in the fetal lung. The placenta also does not co-express the two proteins across the second trimester or at term. INTERPRETATION: This dataset indicates that the lungs are unlikely to be a viable route of SARS-CoV2 fetal infection. The fetal kidney, despite presenting both the proteins required for the infection, is anatomically protected from the exposure to the virus. However, the gastrointestinal tract is likely to be susceptible to infection due to its high co-expression of both proteins, as well as its exposure to potentially infected amniotic fluid. TWEETABLE ABSTRACT: This work provides detailed mechanistic insight into the relative protection & vulnerabilities of the fetus & placenta to SARS-CoV-2 infection by scRNAseq & protein expression analysis for ACE2 & TMPRSS2. The findings help to explain the low rate of vertical transmission.

Mannose receptor (MR) and Toll-like receptor 2 (TLR2) influence phagosome maturation during Leishmania infection.

Leishmania enter macrophages through receptor-mediated phagocytosis and survive the harsh environment of a phagolysosome. Here, we investigated the interaction between mannose receptor (MR), Toll-like receptor 2 (TLR2), and Leishmania, and the subsequent impact on phagosome maturation. Leishmania parasites are able to delay phagosome maturation, not reaching full maturation until 5 hours post-engulfment. Here, maturation of Leishmania major- and Leishmania donovani-containing phagosomes proceeded as expected in the WT macrophages becoming LAMP1 positive by 6 hours. Interestingly, MR-/- macrophages become LAMP1 positive by ~2 hours and ~4 hours post-infection Leishmania-containing phagosomes lost LAMP1 expression and gained the early marker EEA1. LAMP1 expression was again observed by 6 hours. Leishmania LPG was essential for the delay in both WT and MR-/- macrophages but was not essential for the early maturation (2 hours) observed in MR-/- macrophages. Serum opsonization of Leishmania prior to infection induced identical phagosome maturation patterns in WT and MR-/- macrophages. In the absence of MyD88 or TLR2 on macrophages, Leishmania phagosomes matured significantly faster, becoming LAMP1 positive by ~1-2 hours. These studies add to the knowledge that phagosome maturation is influenced by multiple receptor-ligand interactions and signalling pathways.

Resource Allocation to Flight in an Outbreaking Forest Defoliator Malacosoma disstria.

Allocation of larval nutrients affects adult life history traits in insects. This study assessed the effect of moth age and wing loading on flight capacity in an outbreaking forest lepidopteran, Malacosoma disstria Hübner . Insects were collected from high and low density populations after larval feeding, and flight capacity was tested directly with flight mills and indirectly through the allometric relationship between wing area and body size. Insects from these same populations collected as eggs and fed with a synthetic diet in the laboratory were tested in a separate experiment. Male moth propensity to fly increased with wing loading only when moths were collected as pupae after feeding in the field at high population densities. Moth age and wing loading did not affect the distance flown by male moths in any of the population density-nutrient regime combinations tested. Energy use increased with flight distance in both experiments. The slope of the allometric relationship between wing area and body mass did not differ from isometry when moths were collected as pupae after feeding at low and high population densities in the field. The slope of this relationship was steeper for males collected from high than low population densities. There was no allometric relationship between wing area and body mass of moths collected from these same populations as eggs and fed ad libitum in the laboratory as larvae. The results suggest that male M. disstria can allocate resources to different life history traits in response to differences in population density.

NEDD9/Arf6-dependent endocytic trafficking of matrix metalloproteinase 14: a novel mechanism for blocking mesenchymal cell invasion and metastasis of breast cancer.

NEDD9 is an established marker of invasive and metastatic cancers. NEDD9 downregulation has been shown to dramatically reduce cell invasion and metastasis in multiple tumors. The mechanisms by which NEDD9 regulates invasion are largely unknown. In the current study, we have found that NEDD9 is required for matrix metalloproteinase 14 (MMP14) enzymatic recovery/recycling through the late endosomes to enable disengagement of tissue inhibitor of matrix metalloproteinase 2 (TIMP2) and tumor invasion. Depletion of NEDD9 decreases targeting of the MMP14/TIMP2 complex to late endosomes and increases trafficking of MMP14 from early/sorting endosomes back to the surface in a small GTPase ADP ribosylation factor-6 (Arf6)-dependent manner. NEDD9 directly binds to Arf6-GTPase-activating protein, ARAP3 and Arf6-effector GGA3, thereby facilitating the Arf6 inactivation required for MMP14/TIMP2 targeting to late endosomes. Re-expression of NEDD9 or a decrease in Arf6 activity is sufficient to restore MMP14 activity and the invasive properties of tumor cells. Importantly, NEDD9 inhibition by Vivo-Morpholinos, an antisense therapy, decreases primary tumor growth and metastasis in xenograft models of breast cancer. Collectively, our findings uncover a novel mechanism to control tumor-cell dissemination through NEDD9/Arf6-dependent regulation of MMP14/TIMP2 trafficking, and validate NEDD9 as a clinically relevant therapeutic target to treat metastatic cancer.

Diurnal variations in brain iron concentrations in BXD RI mice.

Under normal and dietary iron deficiency conditions, the BXD recombinant inbred (RI) strains of mice show large variations in regional brain iron concentration, particularly in the ventral midbrain (VMB). In a study utilizing just one of the BXD strains, diurnal changes in subregional brain iron concentration were found, which were dependent on the brain region and sex of the mice. The focus of this study was to determine if diurnal changes in VMB can be found across other BXD RI strains and whether a diurnal effect would be common to all strains or variable across strains similar to the large strain variability in iron concentrations determined during the first part of the light phase. Eight RI (BXD type) strains of mice of both sexes were selected for this study. Mice were sacrificed at postnatal day 120: half in the light phase (LP) and half in the dark phase (DP) of the light-dark cycle. Iron concentrations were determined in VMB, which was the primary region of interest, and five other brain regions. Exploratory analysis was also done on liver and spleen iron concentrations to assess for diurnal changes. Three strains showed clear diurnal variation in iron in the VMB and the others strains showed diurnal variations in other regions. These changes were not equally apparent in both sexes. Exploratory analysis also found strain×sex-dependent diurnal differences in spleen and liver iron. In conclusion, significant brain-regional-specific diurnal changes in total iron concentrations were found in a selection of BXD RI mice. Sex and strain are functional determinates of which regions will be affected and in what direction the affect will be. The study provides an animal model for future work into determining the biological and genetic basis of circadian influences on VMB iron homeostasis.

Iron deficiency alters expression of dopamine-related genes in the ventral midbrain in mice.

A clear link exists between iron deficiency (ID) and nigrostriatal dopamine malfunction. This link appears to play an important role in at least restless legs syndrome (RLS) if not several other neurological diseases. Yet, the underlying mechanisms remain unclear. The effects of ID on gene expression in the brain have not been studied extensively. Here, to better understand how exactly ID alters dopamine functioning, we investigated the effects of ID on gene expression in the brain, seeking to identify any potential transcription-based mechanisms. We used six strains of recombinant inbred mice (BXD type) known to differ in susceptibility to ID in the brain. Upon weaning, we subjected mice from each strain to either an iron-deficient or iron-adequate diet. After 100 days of dietary treatment, we measured the effects of ID on gene expression in the ventral midbrain, a region containing the substantia nigra. The substantia nigra is the base of the nigrostriatal dopamine pathway and a region particularly affected by iron loss in RLS. We screened for ID-induced changes in expression, including changes in that of both iron-regulating and dopamine-related genes. Results revealed a number of expression changes occurring in ID, with large strain-dependent differences in the genes involved and number of expression changes occurring. In terms of dopamine-related genes, results revealed ID-induced expression changes in three genes with direct ties to nigrostriatal dopamine functioning, two of which have never before been implicated in an iron-dopamine pathway. These were stromal cell-derived factor 1 (Cxcl12, or SDF-1), a ferritin regulator and potent dopamine neuromodulator, and hemoglobin, beta adult chain 1 (Hbb-b1), a gene recently shown to play a functional role in dopaminergic neurons. The extent of up-regulation of these genes varied by strain. This work not only demonstrates a wide genetic variation in the transcriptional response to ID in the brain, but also reveals two novel biochemical pathways by which iron may potentially alter dopamine function.

Effects of IV iron isomaltoside-1000 treatment on regional brain iron status in an iron-deficient animal.

BACKGROUND: Iron deficiency has been documented to affect human cognitive function and conditions with brain iron compromise such as the restless legs syndrome (RLS). Intravenous (IV) iron treatment is used to reduce iron deficiency but its effects on brain iron are not known. It is not known if IV iron is effective in correcting regional brain iron deficiencies nor if it poses a risk of producing iron overload in some brain regions. Preclinical study of IV iron in the iron-deficient (ID) murine model is needed to evaluate and develop IV iron treatments for brain iron deficiency. METHODS: Response to tail vein injections of iron (iron isomaltoside-1000, dose equivalent to 1000 mg for 75 kg adult) or vehicle were evaluated for ID mice by microdialysis assessing non-transferrin bound (NTB) iron in the ventral midbrain (VMB) and autopsy at 3 and 10 days post-injection assessing iron content in critical brain regions. RESULTS: The ID mice showed marked circadian variation in NTB extracellular iron. After iron injection, NTB iron was rapidly increased in the VMB and then decreased over 12h to the levels observed for vehicle. Regional brain iron content at 3 and 10 days post-injection in the iron- compared to vehicle-treated group showed significantly more iron for the VMB and nucleus accumbens but not for the other regions (i.e. prefrontal cortex, caudate-putamen, cerebellum, and pons), which also did not show decreased iron content with the ID diet. CONCLUSION: Iron isomaltoside-1000 given IV corrects the regional brain iron deficiency in these ID mice without producing iron overload in any of the brain regions studied. This is the first demonstration of effects of IV iron in the brain and it provides a useful preclinical model for this assessment, particularly relevant for developing iron treatments for conditions with problematic iron deficiency, e.g. RLS.

Kin recognition: evidence that humans can perceive both positive and negative relatedness.

The evolution of spite entails actors imposing costs on 'negative' relatives: those who are less likely than chance to share the actor's alleles and therefore more likely to bear rival alleles. Yet, despite a considerable body of research confirming that organisms can recognize positive relatives, little research has shown that organisms can recognize negative relatives. Here, we extend previous work on human phenotype matching by introducing a cue to negative relatedness: negative self-resembling faces, which differ from an average face in the opposite direction to the way an individual's own face differs from the average. Participants made trustworthiness and attractiveness judgements of pairs of opposite-sex positive and negative self-resembling faces. Analyses revealed opposing effects of positive and negative self-resembling faces on trustworthiness and attractiveness judgements. This is the first clear evidence that humans are sensitive to negative relatedness cues, and suggests the potential for the adaptive allocation of spiteful behaviour.

Polymorphism and haplotype structure in River Buffalo (Bubalus bubalis) toll-like receptor 5 (TLR5) coding sequence.

Most of the 160 million river buffalo in the world are in Asia where they are used extensively, both as a food source and for draught power. Only recently have investigations begun exploring the buffalo genome for variation that might influence health and productivity of these economically important animals. This paper describes the sequence variability of the toll-like receptor 5 (TLR5) gene, which recognizes bacterial flagellin and is a key player in the immune system. TLR5 is comprised of a single exon that is 2577 bp and codes 858 amino acids. We examined single-nucleotide polymorphisms (SNPs) located within the coding region. Overall, 17 SNPs were discovered, seven of which are non-synonymous. Our study population yielded four different haplotypes. We examined predicted protein domain structure and found that river buffalo, swamp buffalo, and African Forest buffalo shared the same protein domain structure and are more similar to each other than they are to cattle and American bison, which are similar to each other. PolyPhen 2 analysis revealed one amino acid substitution in the river buffalo population with potential functional significance.

Effects of ethanol and ecstasy on conditioned place preference in the rat.

The club drug ecstasy (3,4-methylenedioxymethylamphetamine or MDMA) is often taken recreationally with ethanol (EtOH). We have shown previously that EtOH potentiates the psychomotor effects of MDMA in rats. More recently, we demonstrated in striatal slices that MDMA produced preferential release of serotonin, but when combined with EtOH, the preferential release shifted to dopamine, raising the possibility that administration of EtOH may increase the reward effect of MDMA. To address this possibility, adult male Long-Evans rats were tested for conditioned place preference following treatment with saline, EtOH (0.75 g/kg), MDMA (6.6 mg/kg) or the combination. The only condition that produced a preference for the compartment associated with the drug was that of the drug combination. The current data are in line with anecdotal reports and one study in humans, indicating that EtOH alters the pharmacological effects of MDMA including self reports of enhanced or prolonged euphoria. Thus, administration of EtOH might increase the risk for compulsive use of MDMA, an issue that warrants further study.

Transient pupil constrictions to faces are sensitive to orientation and species.

Previous studies have reported transient pupil constrictions to basic visual attributes (e.g., color and movement) that are processed along the ventral and the dorsal pathways. Specific cortical areas are activated more for faces than most other types of stimuli, raising the possibility that stimulus-specific transient pupil constrictions might also occur for faces. Such pupil responses may be sensitive to stimulus orientation and species since these parameters have been found to affect electrophysiological and behavioral responses to faces. Here we show transient pupil constrictions to upright human faces that are greater than those to scrambled versions, inverted versions, or macaque monkey faces. Similar to findings from electrophysiological studies, the inversion effect occurred for human faces but not macaque faces. Collectively, our findings show that transient pupil constrictions to faces are sensitive to the same parameters that have been found to influence electrophysiological and behavioral measures of face processing (i.e., orientation and species) and thus reveal a novel, objective, and non-invasive method for studying face perception.

Of mice and men, periodic limb movements and iron: how the human genome informs the mouse genome.

The gene, BTBD9, was recently linked to restless legs syndrome, periodic limb movements and iron status in humans. In a homologous region in mouse, an area containing btbd9 was also identified as being related to iron homeostasis. This finding is important as iron status in brain has been implicated in restless legs syndrome.

Sex-contingent face aftereffects depend on perceptual category rather than structural encoding.

Many studies have used visual adaptation to investigate how recent experience with faces influences perception. While faces similar to those seen during adaptation phases are typically perceived as more 'normal' after adaptation, it is possible to induce aftereffects in one direction for one category (e.g. female) and simultaneously induce aftereffects in the opposite direction for another category (e.g. male). Such aftereffects could reflect 'category-contingent' adaptation of neurons selective for perceptual category (e.g. male or female) or 'structure-contingent' adaptation of lower-level neurons coding the physical characteristics of different face patterns. We compared these explanations by testing for simultaneous opposite after effects following adaptation to (a) two groups of faces from distinct sex categories (male and female) or (b) two groups of faces from the same sex category (female and hyper-female) where the structural differences between the female and hyper-female groups were mathematically identical to those between male and female groups. We were able to induce opposite aftereffects following adaptation between sex categories but not after adaptation within a sex category. These findings indicate the involvement of neurons coding perceptual category in sex-contingent face aftereffects and cannot be explained by neurons coding only the physical aspects of face patterns.

Evidence for adaptive design in human gaze preference.

Many studies have investigated the physical cues that influence face preferences. By contrast, relatively few studies have investigated the effects of facial cues to the direction and valence of others' social interest (i.e. gaze direction and facial expressions) on face preferences. Here we found that participants demonstrated stronger preferences for direct gaze when judging the attractiveness of happy faces than that of disgusted faces, and that this effect of expression on the strength of attraction to direct gaze was particularly pronounced for judgements of opposite-sex faces (study 1). By contrast, no such opposite-sex bias in preferences for direct gaze was observed when participants judged the same faces for likeability (study 2). Collectively, these findings for a context-sensitive opposite-sex bias in preferences for perceiver-directed smiles, but not perceiver-directed disgust, suggest gaze preference functions, at least in part, to facilitate efficient allocation of mating effort, and evince adaptive design in the perceptual mechanisms that underpin face preferences.

Raised salivary testosterone in women is associated with increased attraction to masculine faces.

Women's preferences for masculinity in men's faces, voices and behavioral displays change during the menstrual cycle and are strongest around ovulation. While previous findings suggest that change in progesterone level is an important hormonal mechanism for such variation, it is likely that changes in the levels of other hormones will also contribute to cyclic variation in masculinity preferences. Here we compared women's preferences for masculine faces at two points in the menstrual cycle where women differed in salivary testosterone, but not in salivary progesterone or estrogen. Preferences for masculinity were strongest when women's testosterone levels were relatively high. Our findings complement those from previous studies that show systematic variation in masculinity preferences during the menstrual cycle and suggest that change in testosterone level may play an important role in cyclic shifts in women's preferences for masculine traits.

Disparity in holoprotein/apoprotein ratios of different standards used for immunoquantification of hepatic cytochrome P450 enzymes.

An analysis of reported hepatic abundances of CYP3A4 and 3A5 indicated that values determined by immunoquantification using commercially available, unpurified recombinant enzymes as standards are significantly lower than those determined using purified enzymes or human liver microsomes characterized with lysosomal peptides (CYP3A4: mean 45 versus 121 pmol/mg protein, p < 0.01; CYP3A5: mean 28 versus 83 pmol/mg protein, p < 0.05). When immunoquantifying cytochromes P450 (P450s), it is assumed that the holoprotein (holo)/apoprotein ratio is the same in the samples and the standard. Estimates of holo/apoprotein ratios from data reported for a range of P450s purified from human liver and non-commercial recombinant systems indicated less than complete and variable heme coupling dependent on enzyme and system.

Salience of emotional displays of danger and contagion in faces is enhanced when progesterone levels are raised.

Findings from previous studies of hormone-mediated behavior in women suggest that raised progesterone level increases the probability of behaviors that will reduce the likelihood of disruption to fetal development during pregnancy (e.g. increased avoidance of sources of contagion). Here, we tested women's (N=52) sensitivity to potential cues to nearby sources of contagion (disgusted facial expressions with averted gaze) and nearby physical threat (fearful facial expressions with averted gaze) at two points in the menstrual cycle differing in progesterone level. Women demonstrated a greater tendency to perceive fearful and disgusted expressions with averted gaze as more intense than those with direct gaze when their progesterone level was relatively high. By contrast, change in progesterone level was not associated with any change in perceptions of happy expressions with direct and averted gaze, indicating that our findings for disgusted and fearful expressions were not due to a general response bias. Collectively, our findings suggest women are more sensitive to facial cues signalling nearby contagion and physical threat when raised progesterone level prepares the body for pregnancy.

Facial appearance is a cue to oestrogen levels in women.

Although many accounts of facial attractiveness propose that femininity in women's faces indicates high levels of oestrogen, there is little empirical evidence in support of this assumption. Here, we used assays for urinary metabolites of oestrogen (oestrone-3-glucuronide, E1G) and progesterone (pregnanediol-3-glucuronide, P3G) to investigate the relationship between circulating gonadal hormones and ratings of the femininity, attractiveness and apparent health of women's faces. Positive correlations were observed between late follicular oestrogen and ratings of femininity, attractiveness and health. Positive correlations of luteal progesterone and health and attractiveness ratings were marginally significant. Ratings of facial attributions did not relate to hormone levels for women wearing make-up when photographed. There was no effect of sex of rater on the relationships between oestrogen and ratings of facial appearance. These findings demonstrate that female facial appearance holds detectable cues to reproductive health that are considered attractive by other people.

Menstrual cycle, trait estrogen level, and masculinity preferences in the human voice.

Men with low testosterone (feminine men) invest in relationships and offspring more than men with high testosterone (masculine men). Women's attraction to testosterone dependent traits (e.g. masculine face shape) is enhanced during the late-follicular, fertile phase of the menstrual cycle. Attractive, feminine women have stronger preferences for masculine men as possible long-term partners than less attractive, masculine women. We manipulated 2 testosterone related vocal traits (voice pitch and apparent vocal-tract length) in voices to test if women prefer masculinized men's voices to feminized men's voices; masculinity preferences are enhanced at the fertile (late-follicular) menstrual cycle phase; the amount that masculinity preferences shift cyclically relates to average estrone-3-glucuronide concentration (the primary urinary metabolite of estrone, E3G). We found women displayed general masculinity preferences for men's voices; masculinity preferences were greater in the fertile (late-follicular) phase of the cycle than the non-fertile (early-follicular and luteal) phase; and this effect was most pronounced for women with low average E3G concentration. As feminine women (i.e. those with high average E3G levels) are most able to obtain investment even from masculine men, these women may not need to change their mating preference or strategy during the menstrual cycle as much as masculine women.