COVID-19 and “natural” experiments arising from physical distancing: a hypothetical case study from chronobiology

ABSTRACT

With countless “natural” experiments triggered by the COVID-19-associated physical distancing, one key question comes from chronobiology: “When confined to homes, how does the reduced exposure to natural daylight arising from the interruption of usual outdoor activities plus lost temporal organization ordinarily provided from workplaces and schools affect the circadian timing system (the internal 24 h clock) and, consequently, health of children and adults of all ages?” Herein, we discuss some ethical and scientific facets of exploring such natural experiments by offering a hypothetical case study of circadian biology.     

Behavioral response ofGraminella nigrifrons (Homoptera: Cicadellidae) to experimentally manipulated vibrational signals

Mate recognition for the leafhopper Graminella nigrifrons(Forbes) occurs when a male spontaneously emits a multisectional vibrational calling song to which females respond by emitting simple pulses. Significant differences were found among males in the duration, number of chirps, and chirp rate within sections of the song and the total song. Repeatability (proportion of total variation due to differences among males) of call features ranged from very low (0.04 for total chirps in song) to high (0.67 for section 3 chirp rate). However, song modification and playback experiments revealed that the variation in the measured song features was not important in determining whether a female will respond. Rather, female response depended only on the presence of two of the three types of pulses which comprise a chirp. These essential pulses were found within chirps of all call sections that contain chirps. Manipulation of chirp rates from 0.58 to 2.70 times the normal rate did not affect female response, nor did changing the period of silence between the essential pulse types from 0.25 to 1.75 times the normal period. These results suggest that components of the male calling song function in mate recognition but are not used by females to discriminate among conspecific males.     

Lipolytic and peroxidative enzyme expressions in cultured potato tuber cells

Potato cells (cv. Norchip) were cultured from tuber parenchymal tissue and subcultured to dissociate and habituate the despecialized cells. After several subculturings on a minimal nutrient media, this line of cells demonstrated repeatable physical growth profiles for dry weight (DW), fresh weight (FW) and protein. Two enzymes of plant lipid metabolism were investigated, lipolytic acyl hydrolase (LAH) and lipoxygenase (LOX), which respectively liberate and peroxidize fatty acids from lipid in cellular membranes. LAH, measured as p-nitrophenyl palmitate hydrolase, was present in this line of cells in easily detectable amounts (317 units g^-1 DW) albeit much lower than that found in mother tuber (9878 units g^-1 DW). The presence of LAH in this line is significant because LAH isozymes are often described as storage proteins, yet activity per gram fresh weight in these unorganized cells is reasonably constant until culture growth exits the linear phase. However, LOX, the most active free fatty acid metabolizing enzyme in potato tubers (89,800 units g^-1 DW), was not detectable in this line of callus or suspension cultured cells. The absence of LOX activity in this line of cells was verified by a number of assay approaches and was confirmed by activity staining of extracted enzymes separated in polyacrylamide gels. The absence of LOX in these cultured cells is especially important in determining the functions of this lipid peroxidation system and how it may be genetically regulated.Mention of company or trade name does not imply endorsement by the United States Department of Agriculture over others not named.A laboratory cooperatively operated by the Midwest Area, Agricultural Research Service, U.S. Department of Agriculture, The Minnesota Agricultural Experiment Station, the North Dakota Agrcultural Experiment Station, and the Red River Valley Potato Grower's Association.     

Identification of a developmentally regulated sialidase inEimeria tenella that is immunologically related to theTrypanosoma cruzi enzyme

Sporozoites and merozoites of three species ofEimeria, E. tenella, E. maxima, andE. necatrix, that cause diarrhea in chickens worldwide, were examined for their expression of sialidase (SA) activity. The enzyme was found in three species, and the activity of merozoites was 10–20 times higher than that of sporozoites. The enzyme was resistant to degradation by proteases that are normally present in the intestine, a site inhabited by theEimeria parasites, and it was relatively resistant to heat, with optimum activity being at 40°C, which is within the range of temperature in the chicken intestine (40–43°C).E. tenella SA was immuniprecipitated by monoclonal and polyclonal antibodies raised against theTrypanosoma cruzi SA (TCSA), and enzyme activity was neutralized by these antibodies.E. tenella SA was identified by immunoblots as a doublet of molecular weight 190 000 and 180 000 using, as a probe, anti-TCSA antibodies and antibodies against a synthetic peptide (TR) derived from the long tandem repeat domain of TCSA. Binding of the monoclonal and polyclonal antibodies toE. tenella was completely blocked by TR, but not by an irrelevant peptide (BR). Therefore,E. tenella expresses a developmentally regulated SA that is structurally related to theT. cruzi counterpart. Because of the high SA activity in merozoites, and by analogy with other SA-producing microbes that inhabit mucin-rich epithelia, we suggest that theEimeria SA plays a role in desialylating intestinal mucins to reduce viscosity of the local environment and thereby facilitate parasite migration. The enzyme could also play a role in host cell-parasite interaction.Abbreviations SA sialidase (neuraminidase) - Neu5Ac N-acetylneuraminic acid - 4-MU-Neu5Ac 2-(4-methylumbelliferyl)--N-acetyl-d-neuraminic acid - BSA bovine serum albumin - PBS phosphate buffered saline - PMSF phenylmethylsulfonyl fluoride - PNA peanut agglutinin - Ab antibody - TCN-2 monoclonal antibody toT. cruzi sialidase, anti-Ars, monoclonal antibody top-azophenylarsonate - TCSA Trypanosoma cruzi sialidase     

Ultrastructural and autoradiographic study of the intranuclear inclusions of Pinguicula lusitanica L.

Intranuclear inclusions are found in Pinguicula lusiaanica. Observations with an electron microscope equipped with a goniometric stage demonstrate that they are formed with numerous similarly oriented lamellae. Enzyme digestions of ultrathin sections show that the inclusions are composed of protein. An autoradiographic study, after incubation of pieces of leaves or roots in a physiological medium containing tritiated arginine, shows that the inclusions grow in tissue incubated in such an artificial mineral medium.     

Ultrastructural events and kinetics of microcyst germination in the myxomycete Didymium iridis

Microcysts of the myxomycete Didymium iridis were induced to excyst by transfer to 5mM potassium phosphate buffer. After 1 h in suspension, 90% of the microcysts had germinated into myxamoebae distinguishable by phase contrast microscopy and staining with Lugol's iodine. Both pH and osmolarity affected the kinetics of excystment. The rate and extent of excystment were decreased by cycloheximide but remained unaffected by actinomycin D, suggesting a requirement for protein synthesis but not RNA synthesis. Initially, the outer wall layers separated from the inner layer, which gradually expanded and loosened. The protoplast rehydrated and reverted to a vegetative morphology. Excysting cells were characterized by nucleolar inclusions, changes in the nuclear envelope and plasma membrane, appearance of ringed cisternal elements and microbodies in the cytoplasm, and formation of a densely fibrous zone adjacent to the site of emergence. Excysting populations have been classified into characteristic stages: mature, initiated, swollen, and pre-emergent microcysts.Florida Agricultural Experiment Station Journal Series No. 2407     

Crown gall transformation of lentil (Lens culinaris Medik.) with virulent strains of Agrobacterium tumefaciens

Summary Four diverse strains of Agrobacterium tumefaciens (C58, Ach5, GV3111, and A281) were capable of inducing tumors at a high frequency on inoculated stems of lentil (Lens culinaris Medik. cultivar Laird) in vivo, and on excised shoot apices in vitro. GV3111 and Ach5 produced the largest and heaviest tumors in vivo, while A281 produced the heaviest tumors in vitro. Tumor formation and opine production are indicative of plant cell transformation and tumors produced appropriate opines: nopaline (C58), octopine (Ach5 and GV3111), and agropine and mannopine (A281). Southern analysis of DNA from a tumor line produced by strain C58 showed that a T-DNA fragment had been transferred into the lentil genome.     

A substance released by metamorphosing larvae and young polyps ofHydractinia echinata induces metamorphosis in conspecific larvae

Summary A metamorphosis-inducing factor was isolated from medium conditioned by either metamorphosing larvae or 3-day postmetamorphic primary polyps. The factor has a molecular weight 8 kDa and is heatlabile. It does not induce metamorphosis of isolated posterior fragments and is therefore not identical to the internal signal described by Schwoerer-Böhning et al. (1990). The biological significance of the substance is currently unclear, therefore its inducing activity may be a side effect.     

Synthesis of the intranuclear crystals in Scolopendrium vulgare (L.) SM

Proteinaceous intranuclear crystals are found in the fern Scolopendrium vulgare. During mitosis these crystals are eliminated from the nucleus into the cytoplasm, where they are dissolved. New crystals appear in the nucleus. The site of synthesis of intranuclear crystal proteins was investigated using quantitative ultrastructural autoradiography after incubation with tritiated lysine. The results suggest a migration of cytoplasmic proteins to the nucleus, part of which would then be incorporated into the intranuclear crystals.