Golgi'S Dichromate-Silver Method 3. Chromating Fluids

Slices of cat brains that had been fixed in 10% aqueous formalin for various periods from 2 days to over a year were subjected to different types of chromation prior to impregnation with silver nitrate. Acid solutions of Al, Ba, Ca, Cd, Ce, Co, Cu, Fe, K, Ni, Sr and Zn chromates were tested for usefulness as chromating agents. The chromates of Cd, Co, K, Sr and Zn were found to be best; Ca, Ce and Ni gave positive results, but Al, Ba, Cu and Fe were quite unsatisfactory. Acetic acid was somewhat preferable to formic as the acidifying agent. A formula consisting of potassium chromate, 5% aq., 100 ml. and glacial acetic acid 6-8 ml. was found to be similar in action to comparable mixtures that contained the chromate of Cd, Co, Sr or Zn. Brain slices chromated 24-48 hours in these acidified chromates and silvered in 0.75-1.0% silver nitrate for 12-24 hours at 37-40° C. gave at least three times as many good preparations as similar specimens chromated with plain potassium dichromate solution.     

Topsy-turvy: turning the counter-current heat exchange of leatherback turtles upside down

Counter-current heat exchangers associated with appendages of endotherms feature bundles of closely applied arteriovenous vessels. The accepted paradigm is that heat from warm arterial blood travelling into the appendage crosses into cool venous blood returning to the body. High core temperature is maintained, but the appendage functions at low temperature. Leatherback turtles have elevated core temperatures in cold seawater and arteriovenous plexuses at the roots of all four limbs. We demonstrate that plexuses of the hindlimbs are situated wholly within the hip musculature, and that, at the distal ends of the plexuses, most blood vessels supply or drain the hip muscles, with little distal vascular supply to, or drainage from the limb blades. Venous blood entering a plexus will therefore be drained from active locomotory muscles that are overlaid by thick blubber when the adults are foraging in cold temperate waters. Plexuses maintain high limb muscle temperature and avoid excessive loss of heat to the core, the reverse of the accepted paradigm. Plexuses protect the core from overheating generated by muscular thermogenesis during nesting.     

Year of Birth Effects in the Historical Decline of Tuberculosis Mortality: A Reconsideration

Birth cohort patterns in mortality are often used to infer long-lasting impacts of early life conditions. One of the most widely accepted examples of a birth cohort effect is that of tuberculosis mortality before the late 1940s. However the evidential basis for claims of cohort-specific declines in tuberculosis mortality is very slight. Reanalysis of original or enhanced versions of datasets used previously to support claims of cohort effects in tuberculosis mortality indicated that: 1. where the initial decline in tuberculosis mortality occurred within the period of observation, onset of decline occurred simultaneously in many age groups, in a pattern indicative of ‘period’ not cohort-dependent effects. 2. there was little evidence of ‘proportional hazard’-type cohort patterns in tuberculosis mortality for any female population studied. Therefore any mechanisms proposed to underlie this type of cohort pattern in male mortality must be sex-specific. 3. sex ratios of tuberculosis mortality at older ages peaked in cohorts born around 1900, and resembled cohort sex ratios of lung cancer mortality. This analysis indicates that age-specific patterns in the decline in tuberculosis mortality before 1950 are unlikely to reflect improvements in early life conditions. The patterns observed are generally more consistent with the influence of factors that reduced mortality simultaneously in most age groups. Additional influences, possibly smoking habits, impeded the decline of tuberculosis in older adult males, and produced the sex-specific shifts in age distributions of mortality that were previously interpreted as evidence of cohort-dependent mortality decline.     

Biocontrol of Damping-off Diseases Caused by Rhizoctonia solani and Pythium ultimum with Alginate Prills of Gliocladium virens, Trichoderma hamatum and Various Food Bases

Alginate prills were formulated with the biomass of isolates of Gliocladium virens and Trichoderma spp. and various food bases (wheat bran, corn cobs, peanut hulls, soy fiber, castor pomace, cocoa hulls and chitin). Alginate prills with G. virens (Gl-21) biomass and all food bases except cocoa hull meal significantly reduced the damping-off of zinnia in a soil-less mix caused by Rhizoctonia solani and Pythium ultimum. The prills with bran, soy fiber, castor pomace or chitin resulted in stands similar to those in the non-infested control. In soil, prills with all the food bases and Thrichoderma hamatum (TRI-4) biomass controlled the damping-off of cotton caused by R. solani and gave stands comparable to, or better than, those in the non-infested control soil. Prills with all the food bases resulted in a proliferation of Gl-21 in a soil-less mix and of Gl-21 and TRI-4 in soil. Prills with food bases and TRI-4 biomass reduced the survival of R. solani in infested beet seed to less than 30%, with bran and chitin being the most effective food bases; prills with Gl-21 biomass and all food bases also reduced the survival of R. solani in beet seed, but not as much as did prills with TRI-4 biomass. In prills containing wheat bran, soy fiber or chitin, the biocontrol isolate Th-58 (T. harzianum) was almost as effective as TRI-4, but isolate Gl-3 (G. virens) was less effective. There was no significant interaction between the biocontrol fungus and the food base. The results suggest that the intrinsic properties of a selected fungus isolate are more important than some formulation variables in biocontrol.     

In vitro cytotoxicity evaluation of some substituted isatin derivatives

A range of substituted 1H-indole-2,3-diones (isatins) were synthesized using standard procedures and their cytotoxicity evaluated against the human monocyte-like histiocytic lymphoma (U937) cell line in vitro. SAR studies identified C(5), C(6), and C(7) substitution greatly enhanced activity with some di- and tri-halogenated isatins giving IC(50) values <10 microM. Of the 23 compounds tested, four were selected for further screening against a panel of five human cancer cell lines. These compounds, in general, showed greater selectivity toward leukemia and lymphoma cells over breast, prostate, and colorectal carcinoma cell lines. The most active compound, 5,6,7-tribromoisatin (2p), was found to be antiproliferative at low micromolar concentrations and also activated the effector caspases 3 and 7 in a dose-dependent manner. These results indicate that di- and tri-substituted isatins may be useful leads for anticancer drug development in the future.     

Community structure of Diplostomum spp. (Digenea: Diplostomidae) in eyes of fish: Main determinants and potential interspecific interactions

Parasite communities in freshwater fish vary annually and seasonally and can be influenced by the length, age, sex and phylogeny of hosts, and by associations among parasite species. We assessed the influence of these factors in species of Diplostomum in the eyes of 828 fish in 20 different species collected in a single lake in early summer or autumn over a 5 year interval. Strong negative associations were found between five pairs of Diplostomum spp. and associations were strongest among abundant species. Most interspecific associations occurred among species inhabiting the lens, suggesting competitive interactions. However, the strongest association occurred between two Diplostomum spp. that inhabit different tissues (i.e., the vitreous humour and lens), indicating processes other than direct competition. Seasonal variation was small compared with inter-annual variation. Infection intensities were high in 2006 and decreased dramatically in 2010 and 2011. Infracommunity composition and structure showed no clear correspondence to the ecology or phylogeny of host species. Host length and age, but not sex, had significant effects on the structure of Diplostomum infracommunities in lenses. However, a significant amount of variance in lens infracommunities could not be explained, indicating the potential importance of other factors such as resistance or exposure in determining infracommunity structure.     

Genome sequence of the phage-gene rich marine Phaeobacter arcticus type strain DSM 23566T

Phaeobacter arcticus Zhang et al. 2008 belongs to the marine Roseobacter clade whose members are phylogenetically and physiologically diverse. In contrast to the type species of this genus, Phaeobacter gallaeciensis, which is well characterized, relatively little is known about the characteristics of P. arcticus. Here, we describe the features of this organism including the annotated high-quality draft genome sequence and highlight some particular traits. The 5,049,232 bp long genome with its 4,828 protein-coding and 81 RNA genes consists of one chromosome and five extrachromosomal elements. Prophage sequences identified via PHAST constitute nearly 5% of the bacterial chromosome and included a potential Mu-like phage as well as a gene-transfer agent (GTA). In addition, the genome of strain DSM 23566^T encodes all of the genes necessary for assimilatory nitrate reduction. Phylogenetic analysis and intergenomic distances indicate that the classification of the species might need to be reconsidered.     

Characterization of rat cellular retinol-binding protein II expressed in Escherichia coli

Rat cellular retinol-binding protein II (CRBP II) is a small (15.6 kDa) intracellular protein that binds all-trans-retinol. In the adult rat, expression of the CRBP II gene is essentially limited to the small intestinal lining cells (enterocytes), suggesting that CRBP II may be uniquely adapted for intestinal metabolism of newly absorbed retinol. Functional and structural analysis of this protein has been hampered by difficulties in freeing rat intestinal CRBP II from its ligand without denaturation. To circumvent this problem, we have obtained efficient expression of rat apoCRBP II in Escherichia coli. The purified E. coli-derived apoprotein, when complexed with all-trans-retinol, demonstrates fluorescence excitation-emission spectra and absorption spectra indistinguishable from that of CRBP II-retinol isolated from rat intestine. Quantitative ligand binding studies were performed by monitoring either the fluorescence of bound retinol or the quenching of protein fluorescence. They revealed that E. coli-derived CRBP II binds retinol tightly (the apparent dissociation constant is estimated to be 10(-7)-10(-8) M), with a stoichiometry of 1:1. Fluorescence quenching studies used acrylamide as a probe for the exposure of the 4 tryptophan residues to solvent. The results indicate that although there is heterogeneity in the exposure of these 4 tryptophan residues to solvent, they are situated in a relatively nonpolar environment. These studies suggest that E. coli-derived apoCRBP II will serve as a useful model for studying retinol-protein interactions.