Assessment and comparison of neural morphology through metrical feature extraction and analysis in neuron and neuron–glia cultures

The morphology of dissociated single cerebellar Purkinje cells obtained from wild-type P1 CD1 mice was assessed in the absence and in the presence of glia. A dedicated noninvasive technique based on optical microscopy was developed. Image processing algorithms were implemented to extract metrical features characterizing cell structure and dendritic arborization. The morphological features were analyzed in order to identify quantitative differences in Purkinje cell morphology due to interactions with astrocytes.     

Expression sites and developmental regulation of genes encoding (1→3,1→4)-β-glucanases in germinated barley

Expression sites of genes encoding (13,14)--glucan 4-glucanohydrolase (EC 3.2.1.73) have been mapped in germinated barley grains (Hordeum vulgare L.) by hybridization histochemistry. A³²P-labelled cDNA (copy DNA) probe was hybridized to cryosections of intact barley grains to localize complementary mRNAs. No mRNA encoding (13,14)--glucanase is detected in ungerminated grain. Expression of (13,14)--glucanase genes is first detected in the scutellum after 1 d and is confined to the epithelial layer. At this stage, no expression is apparent in the aleurone. After 2 d, levels of (13,14)--glucanase mRNA decrease in the scutellar epithelium but increase in the aleurone. In the aleurone layer, induction of (13,14)--glucanase gene expression, as measured by mRNA accumulation, progresses from the proximal to distal end of the grain as a front moving away from, and parallel to, the face of the scutellum.Abbreviations cDNA copy DNA - RNase ribonuclease     

Nutritional status and age at secondary sterility in rural Bangladesh

In a prospective study of 2324 women in Matlab, Bangladesh, the occurrence of primary and 2ndary sterility by age groups was examined. The results were related to the nutritional status of the women, as assessed by measurements of height, weight, arm circumference and ponderal index. Approximately 98% of the women who were in the age group 15-19 were found to be fertile. This proportion decreases gradually up to the age group 30-34 years and thereafter declines sharply, reaching only 31% in the age group 45-49. The height data suggest no significant difference in the age pattern of sterility among the 3 groups of women, although there is a slight tendency that women who were less tall reached menopause earlier than the other 2 groups. Variations in weight are more conspicuous than in height. There is the suggestion that thinner women may experience an earlier menopause. Women having an arm circumference less than 21 cm, between 21-22 cm, and 23 cm and above, and currently aged 17 years, have an expected fertile life estimated at 25.0, 25.8, and 26.6 years respectively. The median ages at sterility were 42.8, 44.0, and 44.3 years respectively with a difference of about 1 year between the 1st 2 groups. This suggests that sterility occurs earlier among the thinner women. Since detailed investigation of nutritional status was not possible, it was assessed by anthropometry. There was strong evidence that nutritional status is an important factor in the estimated age at sterility, with thinner women experiencing an earlier menopause. Although it is impossible to measure the onset of sterility, one can obtain a minimum estimate of it from the age-specific distribution of the proportion of women who have not produced a child for 5 years of being at risk.     

Phytoplankton primary production in a mesotrophic pond in sub-tropical Bangladesh

Annual gross primary productivity in mesotrophic Shahidullah Hall pond (Dhaka, Bangladesh) was 1383.35 g C m⁻² y⁻¹ (arithmetic mean). Daily primary productivity (between 1.6 and 6.8 g C m⁻² d⁻¹ was correlated with chlorophylla, day length and dissolved silica. Chlorophylla related significantly withk, incident light, SRP, alkalinity and conductivity. A negative correlation existed between biomass and rainfall. Productivity, biomass, conductivity, alkalinity, and SRP increased after mid-winter.k, I _k andZ _eu varied according seasonally.P _max related directly with temperature. Seasonal variation of ∝ ^B was 0.0049–0.0258 mg C (mg chla mmol PAR)⁻¹ m⁻². Q₁₀ was 2.12, community respiration 1334.99 g C m⁻² y⁻¹, and the underwater light climate 186.43μE m⁻² s⁻¹.     

Immunochemical analysis of the membrane proteins of rat liver and Zajdela hepatoma mitochondria

The contents of mitochondrial inner membrane protein complexes were compared in normal liver and in Zajdela hepatoma mitochondria by the immunotransfer technique. Antibodies against core proteins 1 and 2, cytochrome c1, the iron-sulfur protein of Complex III, subunits I and II of cytochrome oxidase, and the alpha and beta subunits of the F1-ATPase were used. In addition, antibodies against a primary dehydrogenase, beta-hydroxybutyrate dehydrogenase, as well as the outer membrane pore protein were used. The results indicate that the components of the cytochrome chain and porin are greatly enriched in hepatoma mitochondria compared to normal rat liver mitochondria. This enrichment was also reflected in the rates of respiration in tumor mitochondria using a variety of substrates. Enrichment of porin may partially account for increased hexokinase binding to tumor mitochondria. In contrast to the respiratory chain components, the F1-ATPase and F0 (measured by DCCD binding) were not increased in tumor mitochondria. Thus, Zajdela hepatoma mitochondria components are nonstoichiometric, being enriched in oxidative capacity but relatively deficient in ATP synthesizing capacity. Finally, beta-hydroxybutyrate dehydrogenase, which is often decreased in hepatoma mitochondria, was shown here by immunological methods to be decreased by only 40%, whereas enzyme activity was less than 5% of that in normal rat liver.     

Altered metabolic states do not change the intracellular distribution of hexokinase in Zajdela hepatoma ascites cells.

The distribution of hexokinase between bound and soluble forms was studied by digitonin fractionation of Zajdela hepatoma ascites cells maintained under various metabolic conditions. Addition of glucose to Zajdela cells respiring on endogenous substrates induces an immediate inhibition of respiration by 50-60% ( Crabtree effect), and a production of acid due to glycolysis. Acid production decreases abruptly after 60s to 50% of the initial rate. The ATP/ADP ratio is not altered by the addition of glucose or by different rates of glycolysis. The uncoupling agent carbonyl cyanide m-chlorophenylhydrazone decreases the ATP/ADP ratio by 10-fold in cells respiring on endogenous substrate, but has little effect on cells oxidizing glucose. Rapid fractionation of the cells under these various metabolic conditions revealed no change in the distribution of hexokinase. Approx. 75% of hexokinase is bound in all cases, in contrast with lactate dehydrogenase, 95% of which was in the soluble form. Longer-term incubations (to 20 min) revealed only slight (10-15%) increases in soluble hexokinase in cells incubated with glucose. Various metabolic inhibitors had little additional affect on the subcellular distribution of hexokinase. Thus a rapid release of hexokinase from mitochondrial membrane is not a mechanism by which glycolysis is regulated in rapidly growing Zajdela hepatoma.     

Aberrant,ectopic expression of VEGF and VEGF receptors 1 and 2 in malignant colonic epithelial cells. Implications for these cells growth via an autocrine mechanism

Vascular endothelial growth factor A (referred to as VEGF) is implicated in colon cancer growth. Currently, the main accepted mechanism by which VEGF promotes colon cancer growth is via the stimulation of angiogenesis, which was originally postulated by late Judah Folkman. However, the cellular source of VEGF in colon cancer tissue; and, the expression of VEGF and its receptors VEGF-R1 and VEGF-R2 in colon cancer cells are not fully known and are subjects of controversy.     

Modular bioreactor for primary human hepatocyte culture: medium flow stimulates expression and activity of detoxification genes

Down-regulation of detoxification genes, notably cytochrome P450 (CYPs), in primary hepatocyte cultures is a long-standing and major concern. We evaluated the influence of medium flow in this model. Hepatocytes isolated from 12 different liver donors were cultured either in a multichamber modular bioreactor (MCmB, flow rate 250-500 μL/min) or under standard/static conditions, and the expression of 32 genes, enzyme activities and biological parameters were measured 7-21 days later. mRNA expression of genes involved in xenobiotic/drug metabolism and transport, including CYP1A1, 1A2, 2B6, 2C9, 3A4 (and activities for some of them), UDP-glucuronosyltransferase (UGT) 1A1, UGT2B4, UGT2B7, glutathione S-transferase (GSTα), and multidrug resistance protein 1 (MDR1) and MRP2, were specifically up-regulated by medium flow as compared with static controls in all cultures tested. In 2-week-old cultures, expression of detoxification genes reached levels close to or higher than those measured in freshly isolated hepatocytes. In contrast, CYP2D6 and most of other tested genes were not affected by medium flow. We conclude that medium flow specifically interferes with, and up-regulates, the activity of xenosensors and/or the expression of detoxification genes in primary human hepatocytes. Down-regulation of detoxification genes in conventional (static) cultures is therefore partly a consequence of the absence of medium circulation.     

A Review of the Potency of Plant Extracts and Compounds from Key Families as an Alternative to Synthetic Nematicides: History,Efficacy, and Current Developments

The global nematicides market is expected to continue growing. With an increasing demand for synthetic chemical-free organic foods, botanical nematicides are taking the lead as replacements. Consequently, in the recent years, there have been vigorous efforts towards identification of the active secondary metabolites from various plants. These include mostly glucosinolates and their hydrolysis products such as isothiocyanates; flavonoids, alkaloids, limonoids, quassinoids, saponins, and the more recently probed essential oils, among others. And despite their overwhelming potential, variabilities in quality, efficacy, potency and composition continue to persist, and commercialization of new botanical nematicides is still lagging. Herein, we have reviewed the history of botanical nematicides and regional progresses, the potency of the identified phytochemicals from the key important plant families, and deciphered some of the impediments involved in standardization of the active compounds in addition to the concerns over the safety of the purified compounds to non-target microbial communities.