A New Technique Facilitating Studies of Scant Cell Specimens

A simple method is described by which multiple cytological and cytochemical studies can be done on a clinical sample that contains relatively few cells. The cells are concentrated by centrifugation. The cell pellet is fixed, frozen and embedded in plastic. Thin (2-μm) sections are cut from the plastic. Thus, each cell may appear in several sections and many slides can be made from a single specimen. The advantages of this method over cytospins and Millipore filter preparations of cell suspensions are optimal utilization of all cells, excellent morphological and immunological preservation and ease and reproducibility of this technique.     

Induction of somatopause in adult mice compromises bone morphology and exacerbates bone loss during aging

Somatopause refers to the gradual declines in growth hormone (GH) and insulin‐like growth factor‐1 throughout aging. To define how induced somatopause affects skeletal integrity, we used an inducible GH receptor knockout (iGHRKO) mouse model. Somatopause, induced globally at 6 months of age, resulted in significantly more slender bones in both male and female iGHRKO mice. In males, induced somatopause was associated with progressive expansion of the marrow cavity leading to significant thinning of the cortices, which compromised bone strength. We report progressive declines in osteocyte lacunar number, and increases in lacunar volume, in iGHRKO males, and reductions in lacunar number accompanied by ~20% loss of overall canalicular connectivity in iGHRKO females by 30 months of age. Induced somatopause did not affect mineral/matrix ratio assessed by Raman microspectroscopy. We found significant increases in bone marrow adiposity and high levels of sclerostin, a negative regulator of bone formation in iGHRKO mice. Surprisingly, however, despite compromised bone morphology, osteocyte senescence was reduced in the iGHRKO mice. In this study, we avoided the confounded effects of constitutive deficiency in the GH/IGF‐1 axis on the skeleton during growth, and specifically dissected its effects on the aging skeleton. We show here, for the first time, that induced somatopause compromises bone morphology and the bone marrow environment.     

Biomechanical consequences of rapid evolution in the polar bear lineage

The polar bear is the only living ursid with a fully carnivorous diet. Despite a number of well-documented craniodental adaptations for a diet of seal flesh and blubber, molecular and paleontological data indicate that this morphologically distinct species evolved less than a million years ago from the omnivorous brown bear. To better understand the evolution of this dietary specialization, we used phylogenetic tests to estimate the rate of morphological specialization in polar bears. We then used finite element analysis (FEA) to compare the limits of feeding performance in the polar bear skull to that of the phylogenetically and geographically close brown bear. Results indicate that extremely rapid evolution of semi-aquatic adaptations and dietary specialization in the polar bear lineage produced a cranial morphology that is weaker than that of brown bears and less suited to processing tough omnivorous or herbivorous diets. Our results suggest that continuation of current climate trends could affect polar bears by not only eliminating their primary food source, but also through competition with northward advancing, generalized brown populations for resources that they are ill-equipped to utilize.     

Regulation of manganese uptake in Synechocystis 6803 by RfrA,a member of a novel family of proteins containing a repeated five-residues domain

The rfrA gene was identified in a suppressor screen of a Synechocystis sp. PCC 6803 strain deficient in both mntC, encoding a component of an ABC transport system for manganese, and psbO, encoding the extrinsic manganese stabilizing protein of photosystem II (PSII). A spontaneous suppressor mutant (DeltaCDeltaO rfrA-Sup) has a point mutation in rfrA, which restores photosynthetic activity to the DeltamntCDeltapsbO double mutant. Manganese transport and photosynthesis are related in that manganese is essential to the function of PSII, and the state of cellular manganese availability influences the rate of oxygen evolution mediated by PSII. Oxygen evolution experiments with the DeltaCDeltaO rfrA-Sup mutant revealed that the mechanism of suppression is not through a direct modification of PSII. Instead, radioactive manganese uptake experiments indicated that RfrA is a regulator of a high affinity manganese transport system different from the more thoroughly characterized manganese ABC transport system in Synechocystis 6803. RfrA was named for the repeated five-residues domain in the amino terminus of the protein. The RFR domain defines a 16-member family in Synechocystis 6803. Predicted proteins with RFR domains have also been identified in other organisms, but RfrA is the first member of this family to be linked to a physiological process.     

Building a Beetle: How Larval Environment Leads to Adult Performance in a Horned Beetle

The link between the expression of the signals used by male animals in contests with the traits which determine success in those contests is poorly understood. This is particularly true in holometabolous insects such as horned beetles where signal expression is determined during metamorphosis and is fixed during adulthood, whereas performance is influenced by post-eclosion feeding. We used path analysis to investigate the relationships between larval and adult nutrition, horn and body size and fitness-related traits such as strength and testes mass in the horned beetle Euoniticellus intermedius. In males weight gain post-eclosion had a central role in determining both testes mass and strength. Weight gain was unaffected by adult nutrition but was strongly correlated with by horn length, itself determined by larval resource availability, indicating strong indirect effects of larval nutrition on the adult beetle’s ability to assimilate food and grow tissues. Female strength was predicted by a simple path diagram where strength was determined by eclosion weight, itself determined by larval nutrition: weight gain post-eclosion was not a predictor of strength in this sex. Based on earlier findings we discuss the insulin-like signalling pathway as a possible mechanism by which larval nutrition could affect adult weight gain and thence traits such as strength.