A benign approach to the preparation of freshwater bryozoan statoblasts for scanning electron microscopy (SEM) imaging

Abstract

Several different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanella and Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals.     

Acetoacetate reduces growth and ATP concentration in cancer cell lines which over-express uncoupling protein 2

Background  

Recent evidence suggests that several human cancers are capable of uncoupling of mitochondrial ATP generation in the presence of intact tricarboxylic acid (TCA) enzymes. The goal of the current study was to test the hypothesis that ketone bodies can inhibit cell growth in aggressive cancers and that expression of uncoupling protein 2 is a contributing factor. The proposed mechanism involves inhibition of glycolytic ATP production via a Randle-like cycle while increased uncoupling renders cancers unable to produce compensatory ATP from respiration.     

The use of adjunctive GPIIb/IIIa inhibitors in patients with unstable angina/non-Q-wave MI undergoing percutaneous coronary intervention

Glycoprotein IIb/IIIa receptor inhibitors represent a relatively new therapeutic approach in the field of antiplatelet therapy. Following the development of abciximab a number of small molecule GPIIb/IIIa inhibitors have been introduced such as tirofiban and eptifibatide. In this fast-moving field the interventional cardiologist needs a framework to guide decision-making for the individual patient. This review covers the efficacy and safety data from the clinical trials of GPIIb/IIIa inhibitors in the context of patients undergoing percutaneous coronary intervention for unstable angina/non-Q-wave myocardial infarction. There is an increasing body of evidence to support the efficacy of GPIIb/IIIa inhibitors in reducing the risk of adverse ischemic events in high and low risk patients undergoing percutaneous coronary intervention. A number of unresolved efficacy and safety issues remain, including the duration of treatment before and after intervention; whether a reduction in the heparin dose would further decrease the risk of hemorrhage without affecting the periprocedural thrombotic rate in patients undergoing PTCA with adjunctive GPIIb/IIIa inhibitors; and the cost-effectiveness of this therapy. When a thorough analysis of cost-effectiveness has been made, it will be easier to advocate the widespread use of these agents in all patients undergoing coronary intervention.     

Motherhood and the hormones of pregnancy modify concentrations of hippocampal neuronal dendritic spines

Short-term fluctuations in steroid hormones such as estradiol (E2) and progesterone (P) can affect the concentration of hippocampal dendritic spines in adult, cycling nulliparous female rats. Pregnancy is characterized by a significantly longer duration of substantially elevated E2 and P compared to the estrous cycle. Thus, even greater changes than those reported during estrus may be evident. In two experiments, we examined the extent to which reproductive and hormonal state altered the concentration of apical neuronal dendritic spines of the CA1 region of the hippocampus in the following age-matched groups (N's = 7-10/group) of rats: in Exp. 1., CA1 dendritic spine density was examined in nulliparous diestrus (DES), proestrus (PRO), and estrus (ES) females, and late-pregnant (LP) (day 21) and lactating (day 5-6; LACT) females. In Exp. 2, the effects on spine density of a regimen mimicking pregnancy (and that stimulates maternal behavior) were examined, using ovariectomized, no hormone-exposed (OVX-minus) vs. sequential P&E(2)-treated (OVX + P&E2) groups. For both experiments, brains were removed, Golgi-Cox-stained and the most lateral tertiary branches of the apical dendrite of completely-stained hippocampal CA1 pyramidal neurons were traced with oil-immersion at x 1600 and dendritic spine density (# spines/10 micro dendritic segment) recorded. In Exp. 1, spine density was increased in LP and LACT females (which were not different) compared to the other virgin groups, including PRO females, who had more spines than DES and ES. In Exp. 2, OVX + P&E2 displayed significantly more dendritic spines per 10 micro than OVX-minus females (and had numbers that were similar to those of LP and LACT from Exp. 1). Pregnancy and its attendant hormonal fluctuations, therefore, may alter hippocampal neurons that regulate some non-pup-directed components of maternal behavior (e.g., nest building) or behaviors that support maternal behavior (e.g., foraging, associative memory).     

Impact of the changing ecology on intertidal polychaetes in an anthropogenically stressed tropical creek,India

Polychaete assemblages and associated environment of 12 strategically selected intertidal stations along the extremely polluted Thane creek on the west coast of India were studied monthly for a year and compared with past available data to investigate changes in the creek ecology due to various anthropogenic activities like industrial, domestic, and solid waste disposal along with land reclamation. Shannon’s index (H′) varied spatially from 0.4 to 1.5, Margalef richness index (d) from 0.4 to 1.1, and evenness index (J) from 0.3 to 0.7 indicating poor polychaete diversity. Ceratonereis burmensis and Lycastis indica were the most abundant and omnipresent polychaetes in the creek indicating their tolerance and adaptability to various degrees of pollution. Hydro-sedimentological investigations revealed enhancement of total nitrogen (TN) and organic carbon (C_org) load and hypoxic levels of dissolved oxygen (DO) over the years. Silt component of sediment was increasing, with proportionate decrease in clay due to various anthropogenic disturbances. Species richness was correlated positively with clay and negatively with silt. The BIO-ENV analyses indicated the strong influence of NO₃ ⁻-N, clay and TN on the distribution patterns of polychaetes. Pollution-tolerant polychaetes like Lycastis ounaryensis and Polydora tentaculata were getting replaced by more pollution-resistant species like C. burmensis in the creek due to changing sediment texture, reduced oxygen levels, and increased C_org and TN.     

Dendrochronological analyses and climatic signals of Alchornea triplinervia in subtropical forest of southern Brazil

The predominance of secondary forest‐species in Brazilian subtropical forests highlights the importance of understanding the ecology of these taxa, and dendrochronology provides valuable information about the growth and climate response of tree species. The wide distribution of Alchornea triplinervia (Spreng.) Mull. Arg. (Euphorbiaceae), and the presence of growth rings in its wood, leds to its selection for this study. Samples were collected from 34 trees growing in rainy dense forest fragments in the cities of Blumenau and Brusque, in southern Brazil, and subjected to standard dendrochronological techniques. The local chronology spanned from 1889 to 2013 and the age of the trees ranged from 32 to 125 years. The mean age was 73 years and annual increment was 1.13 mm year^?1; the diameter and age showed a weak correlation. The results suggest that A. triplinervia growth responded negatively to hot summers and increased rainfall in the previous spring and winter. The high perdiodicity trend characteristic of ENSO activity was reflected in wavelet power spectra of A. triplinervia chronology, affecting positively its growth after strong ENSO events (1970–2009). Negative influences occurred before 1970 during normal ENSO events. Also, there is evidence of the existence of a connection between Pacific and Atlantic sea surface temperatures influencing on tree growth.