The Giardia ventrolateral flange is a lamellar membrane protrusion that supports attachment

Attachment to the intestinal epithelium is critical to the lifestyle of the ubiquitous parasite Giardia lamblia. The ventrolateral flange is a sheet-like membrane protrusion at the interface between parasites and attached surfaces. This structure has been implicated in attachment, but its role has been poorly defined. Here, we identified a novel actin associated protein with putative WH2-like actin binding domains we named Flangin. Flangin complexes with Giardia actin (GlActin) and is enriched in the ventrolateral flange making it a valuable marker for studying the flanges’ role in Giardia biology. Live imaging revealed that the flange grows to around 1 μm in width after cytokinesis, then remains uniform in size during interphase, grows in mitosis, and is resorbed during cytokinesis. A flangin truncation mutant stabilizes the flange and blocks cytokinesis, indicating that flange disassembly is necessary for rapid myosin-independent cytokinesis in Giardia. Rho family GTPases are important regulators of membrane protrusions and GlRac, the sole Rho family GTPase in Giardia, was localized to the flange. Knockdown of Flangin, GlActin, and GlRac result in flange formation defects. This indicates a conserved role for GlRac and GlActin in forming membrane protrusions, despite the absence of canonical actin binding proteins that link Rho GTPase signaling to lamellipodia formation. Flangin-depleted parasites had reduced surface contact and when challenged with fluid shear force in flow chambers they had a reduced ability to remain attached, confirming a role for the flange in attachment. This secondary attachment mechanism complements the microtubule based adhesive ventral disc, a feature that may be particularly important during mitosis when the parental ventral disc disassembles in preparation for cytokinesis. This work supports the emerging view that Giardia’s unconventional actin cytoskeleton has an important role in supporting parasite attachment.     

Synthesis of myelin glycosphingolipids (galactosylceramide and galactosyl(3-O-sulfate)ceramide (sulfatide)) by cloned cell lines derived from mouse neurotumors

Clonal cell lines derived from both spontaneous and chemically induced rat and mouse brain tumors were screened for their ability to incorporate H232SO4 into galactosyl(3-O-sulfate)ceramide (sulfatide). High levels of 35SO4 incorporation into sulfatide were found only in two of the mouse cell lines studied (G26-20 and -24). Tumors produced by subcutaneous injection of these cell lines into C57BL/6 mice were also unique in that they contained high levels of both sulfatide and galactosylceramide. The synthesis of large amounts of sulfatide and galactosylceramide by a clonal cell line of neurological origin suggests that the original tumor was of oligodendrocyte or Schwann cell origin. In common with a large number of mouse and rat astrocyte cell strains and their derived tumors, these glial cells lacked the ability to synthesize gangliosides such as monosialotetraglycosylceramide and disialotetraglycosylceramide (as judged by analytical and [3H]GlcNH2 incorporation studies). This appears to be a unique characteristic of neuroblastoma-derived cell strains such as N18, NB2a, and NB41A.     

Thermoregulation and heterothermy in some of the smaller flying foxes (Megachiroptera) of New Guinea

Summary Body temperature, heterothermy, oxygen consumption, heart rate, and evaporative water loss were studied in four species of flying foxes (Megachiroptera), Dobsonia minor, Nyctimene major, Nyctimene albiventer, and Paranyctimene raptor, from the vicinity of Madang on the north coast of New Guinea.The thermoregulatory response of D. minor resembled that of most other placental mammals weighing 80 to 100 g. Body temperatures were relatively stable at ambient temperatures between 5 and 34°. The mean oxygen consumption at rest between 30 and 35° was 1.26 cc O₂ (g·hr)^–1. At ambient temperatures between 5 and 35° evaporative water loss averaged 4.5 mg (g·hr)^–1 and increased sharply at higher temperatures. When subjected to heat stress the animals panted, salivated, and licked the wings, belly, and uropatagium. At temperatures above 38° the ratio of heat lost through evaporation to heat production exceeded 1. Minimal heart rates in resting animals near thermal neutrality were approximately 275/min.In those parameters measured, N. major which weighed about 80 g resembled D. minor. Nyctimene albiventer and P. raptor weigh less than 30 g and are among the smallest of the flying foxes. Each shows both homeothermic and heterothermic patterns of response. At an ambient temperature of 35° the minimal oxygen consumption of homeothermic N. albiventer and P. raptor were 1.43 and 1.38 cc O₂ (g·hr)^–1, respectively. Oxygen consumption of homeothermic N. albiventer at 25°, 2.59 cc O₂ (g.hr)^–1, was almost quadruple that of torpid animals at the same temperature. During the daytime both N. albiventer and P. raptor characteristically allowed their body temperatures to fall to near 25°. Both readily aroused from the hypothermic state through physiological means. Heart rates of homeothermic N. albiventer resting at 35° ranged from 312 to 326/min while those of animals torpid at 25° were 88 to 96/min.The capacity for heterothermy has not previously been demonstrated in any members of the Megachiroptera, but our data indicate that it can occur on a daily basis in N. albiventer and P. raptor. This capacity appears to be related to size since it occurs in none of the larger flying foxes so far studied.The data presently available indicate that the relation of body weight to standard metabolism in the Megachiroptera is similar to that of the other placental mammals. In the species we studied, thermal conductances were higher, and heart rates, lower than predicted for mammals of their sizes.These studies were carried out during the 1969 Alpha Helix Expedition to New Guinea and were supported in part by grants GB-5139, GB-3656, and GB-8445 from the U. S. National Science Foundation.     

Exploring movement patterns and changing distributions of baleen whales in the western North Atlantic using a decade of passive acoustic data

Six baleen whale species are found in the temperate western North Atlantic Ocean, with limited information existing on the distribution and movement patterns for most. There is mounting evidence of distributional shifts in many species, including marine mammals, likely because of climate‐driven changes in ocean temperature and circulation. Previous acoustic studies examined the occurrence of minke (Balaenoptera acutorostrata) and North Atlantic right whales (NARW; Eubalaena glacialis). This study assesses the acoustic presence of humpback (Megaptera novaeangliae), sei (B. borealis), fin (B. physalus), and blue whales (B. musculus) over a decade, based on daily detections of their vocalizations. Data collected from 2004 to 2014 on 281 bottom‐mounted recorders, totaling 35,033 days, were processed using automated detection software and screened for each species' presence. A published study on NARW acoustics revealed significant changes in occurrence patterns between the periods of 2004–2010 and 2011–2014; therefore, these same time periods were examined here. All four species were present from the Southeast United States to Greenland; humpback whales were also present in the Caribbean. All species occurred throughout all regions in the winter, suggesting that baleen whales are widely distributed during these months. Each of the species showed significant changes in acoustic occurrence after 2010. Similar to NARWs, sei whales had higher acoustic occurrence in mid‐Atlantic regions after 2010. Fin, blue, and sei whales were more frequently detected in the northern latitudes of the study area after 2010. Despite this general northward shift, all four species were detected less on the Scotian Shelf area after 2010, matching documented shifts in prey availability in this region. A decade of acoustic observations have shown important distributional changes over the range of baleen whales, mirroring known climatic shifts and identifying new habitats that will require further protection from anthropogenic threats like fixed fishing gear, shipping, and noise pollution.     

Parallels,differences and lessons: a comparison of the management of foot-and-mouth disease and COVID-19 using UK 2001/2020 as points of reference

Foot-and-mouth disease (FMD) is an extremely infectious viral infection of cloven-hoofed animals which is highly challenging to control and can give rise to national animal health crises, especially if there is a lack of pre-existing immunity due to the emergence of new strains or following incursions into disease-free regions. The 2001 FMD epidemic in the UK was on a scale that initially overwhelmed the national veterinary services and was eventually controlled by livestock lockdown and slaughter on an unprecedented scale. In 2020, the rapid emergence of COVID-19 has led to a human pandemic unparalleled in living memory. The enormous logistics of multi-agency control efforts for COVID-19 are reminiscent of the 2001 FMD epidemic in the UK, as are the use of movement restrictions, not normally a feature of human disease control. The UK experience is internationally relevant as few countries have experienced national epidemic crises for both diseases. In this review, we reflect on the experiences and lessons learnt from UK and international responses to FMD and COVID-19 with respect to their management, including the challenge of preclinical viral transmission, threat awareness, early detection, different interpretations of scientific information, lockdown, biosecurity behaviour change, shortage of testing capacity and the choices for eradication versus living with infection. A major lesson is that the similarity of issues and critical resources needed to manage large-scale outbreaks demonstrates that there is benefit to a ‘One Health’ approach to preparedness, with potential for greater cooperation in planning and the consideration of shared critical resources.     

Secondary (iso)BAs cooperate with endogenous ligands to activate FXR under physiological and pathological conditions

IsoBAs, stereoisomers of primary and secondary BAs, are found in feces and plasma of human individuals. BA signaling via the nuclear receptor FXR is crucial for regulation of hepatic and intestinal physiology/pathophysiology. Aim: Investigate the ability of BA-stereoisomers to bind and modulate FXR under physiological/pathological conditions. Methods: Expression-profiling, luciferase-assays, fluorescence-based coactivator-association assays, administration of (iso)-BAs to WT and cholestatic mice. Results: Compared to CDCA/isoCDCA, administration of DCA/isoDCA, UDCA/isoUDCA only slightly increased mRNA expression of FXR target genes; the induction was more evident looking at pre-mRNAs. Notably, almost 50% of isoBAs were metabolized to 3-oxo-BAs within 4 h in cell-based assays, making it difficult to study their actions. FRET-based real-time monitoring of FXR activity revealed that isoCDCA>CDCA stimulated FXR, and isoDCA and isoUDCA allowed fully activated FXR to be re-stimulated by a second dose of GW4064. In vivo co-administration of a single dose of isoBAs followed by GW4064 cooperatively activated FXR, as did feeding of UDCA in a background of endogenous FXR ligands. However, in animals with biliary obstruction and concomitant loss of intestinal BAs, UDCA was unable to increase intestinal Fgf15. In contrast, mice with an impaired enterohepatic circulation of BAs (Asbt?/?, Ostα?/?), administration of UDCA was still able to induce ileal Fgf15 and repress hepatic BA-synthesis, arguing that UDCA is only effective in the presence of endogenous FXR ligands. Conclusion: Secondary (iso)BAs cooperatively activate FXR in the presence of endogenous BAs, which is important to consider in diseases linked to disturbances in BA enterohepatic cycling.