Single cell biology beyond the era of antibodies: relevance,challenges, and promises in biomedical research

Research of the past two decades has proved the relevance of single cell biology in basic research and translational medicine. Successful detection and isolation of specific subsets is the key to understand their functional heterogeneity. Antibodies are conventionally used for this purpose, but their relevance in certain contexts is limited. In this review, we discuss some of these contexts, posing bottle neck for different fields of biology including biomedical research. With the advancement of chemistry, several methods have been introduced to overcome these problems. Even though microfluidics and microraft array are newer techniques exploited for single cell biology, fluorescence-activated cell sorting (FACS) remains the gold standard technique for isolation of cells for many biomedical applications, like stem cell therapy. Here, we present a comprehensive and comparative account of some of the probes that are useful in FACS. Further, we illustrate how these techniques could be applied in biomedical research. It is postulated that intracellular molecular markers like nucleostemin (GNL3), alkaline phosphatase (ALPL) and HIRA can be used for improving the outcome of cardiac as well as bone regeneration. Another field that could utilize intracellular markers is diagnostics, and we propose the use of specific peptide nucleic acid probes (PNPs) against certain miRNAs for cancer surgical margin prediction. The newer techniques for single cell biology, based on intracellular molecules, will immensely enhance the repertoire of possible markers for the isolation of cell types useful in biomedical research.     

Why history matters to philosophy of physics

Naturalized metaphysics remains the default presupposition of much contemporary philosophy of physics. As metaphysics is supposed to concern the general structure of reality, so scientific naturalism draws upon our best physical theories to attempt to answer the foundational question “par excellence ” viz., “how could the world possibly be the way this theory says it is?” A particular case study, Hilbert's attempt to analyze and explain a seeming “pre-established harmony” between mind and nature, is offered as a salutary reminder that naturalism's ready inference from physical theory to ontology may be too quick.     

Ablation of cyclase-associated protein 2 (CAP2) leads to cardiomyopathy

Cyclase-associated proteins are highly conserved proteins that have a role in the regulation of actin dynamics. Higher eukaryotes have two isoforms, CAP1 and CAP2. To study the in vivo function of CAP2, we generated mice in which the CAP2 gene was inactivated by a gene-trap approach. Mutant mice showed a decrease in body weight and had a decreased survival rate. Further, they developed a severe cardiac defect marked by dilated cardiomyopathy (DCM) associated with drastic reduction in basal heart rate and prolongations in atrial and ventricular conduction times. Moreover, CAP2-deficient myofibrils exhibited reduced cooperativity of calcium-regulated force development. At the microscopic level, we observed disarrayed sarcomeres with development of fibrosis. We analyzed CAP2’s role in actin assembly and found that it sequesters G-actin and efficiently fragments filaments. This activity resides completely in its WASP homology domain. Thus CAP2 is an essential component of the myocardial sarcomere and is essential for physiological functioning of the cardiac system, and a deficiency leads to DCM and various cardiac defects.     

Dam-forming cacti and nitrogen enrichment in a pi?on-juniper woodland in northwestern Arizona

In a pi?on-juniper woodland in northwestern Arizona, connected basal cladodes of a prickly pear cactus ( Opuntia littoralis var. martiniana ) form check dams that cause deposition of N-rich detritus interspaces otherwise lacking litter. Seventy-eight percent of connected basal cladodes measured in transects grew at an angle (with respect to the slope contour) ≤ 45° -an orientation facilitating deposition of flood-borne debris. Soil total N was significantly greater ( P P = 0.0001) compared to adjacent interspaces. Soil total N and organic C above cactus dams were equal to areas beneath canopies (tree and shrub combined). Net NO 3 - (0-5 cm depth) above cactus dams was significantly greater ( P = 0.0001) than below cactus dams, at interspaces, and beneath canopies. Net NH 4 + (0-5 cm soil depth) above cactus dams was significantly greater ( P s ￣x = 0.625, P = 0.0001).     

Variation in Lewsia kelloggii (Portulacaceae) with description of a new species endemic to Idaho

Lewisia kelloggii has been understood as a rare plant with a disjunct range in California and Idaho. Examination of herbarium specimens and analysis of isozymes in 6 Idaho and 7 California populations revealed consistent differences between plants of the 2 states. Fixed differences in alleles at 2 loci (AAT2 and PGI1) distinguished Idaho from California plants. Genetic identities based on isozymes between Idaho and California populations averaged 0.58, lower than the average for congeneric plant species. Idaho plants were smaller than most California plants, but California plants were variable. The most consistent morphological difference between Idaho and California specimens was the difference in the number of glands on the margins of bracts and sepals. Idaho plants had 0 (-5) pink glands on each margin of these organs, all on teeth near the tips. In California plants these organs had 12-25 glands on each margin, the distal ones elevated on teeth and the proximal ones sessile. We recognize the Idaho plants as a new species, L. sacajaweana , and retain the name L. kelloggii for the California populations.     

Fall diet of blue grouse in Oregon

The early fall diet of Oregon blue grouse ( Dendragapus obscurus pallidus ) from Wallowa County, Oregon, was determined from 145 crops obtained during 1981 and 1982. Of more than 50 plant and animal foods in the diet, short-horned grasshoppers ( Acrididae ), prickly lettuce ( Lactuca serriola ), yellow salsify ( Tragopogon dubius ), wild buck-wheat ( Eriogonum spp.), and snowberry ( Symphoricarpos albus ) occurred in 30% or more of the crops and collectively amounted to 68% of the diet by weight. Seven of the 12 most common foods were consumed differentially by the four sex and age classes of birds. Results indicated that blue grouse foraged in forest and grassland habitats.     

A small carnivore survey technique

Thirty-nine previously unpublished reports of the endangered black-footed ferret from Wyoming are listed with dates, locations, number of animals, sources, and comments.     

Influences of sex and weather on migration of mule deer in California

I examined differences by sex and influences of weather on timing and patterns of migration of Rocky Mountain mule deer ( Odocoileus h. hemionus ) in the eastern Sierra Nevada, California, during 1984-87. Deer initiated spring migration from the winter range at about the same time in all years and made extensive use of holding areas at intermediate elevations. Radio-telemetered deer showed strong fidelity to summer ranges over as many as four years. Fall weather produced different patterns of fall migration. Storms during October produced a pulsed migration, in which most animals mmiiggiraatteedd to the winter range during or soon after the storm; in a year without a storm, fall migration was gradual. Despite the influence of storms on the pattern of fall migration, the median date of fall migration by females did not vary over years; however, among males it was later in a year without fall storms.