Moving towards a better understanding of chemotaxis

Eukaryotic cells are thought to move across supporting surfaces through a combination of coordinated processes: polarisation; extension of dynamic protrusions from a leading edge; adhesion-associated stabilisation of some protrusions; centripetal pulling against those leading adhesions; and de-adhesion at the rear. Gradients of extracellular ligands can be detected by cells and then used to guide them either towards the source (in the case of a chemoattractant) or away from the source (in the case of a chemorepellent)--such migration is termed chemotaxis. Recent work suggests that chemotaxis probably emerges from the ability of cells to spatially encode extracellular gradients of ligands, a process for which phosphoinositide 3'-kinase (PI3K) signals alone are insufficient, and to use that vectorial information to bias movement by enhancing the survival, and not the formation, of the protrusions that experience the greatest stimulation.     

Glycolipid-cholesterol monolayers: towards a better understanding of the interaction between the membrane components

In this work, the interaction between a synthetic analog of archaeal lipids and cholesterol was studied using Langmuir technique. The lipid, β-Mal(3)O(C(16+4))(2), contained phytanyl chains attached via two ether bonds to the sn-2 carbon of the glycerol backbone. The preliminary studies showed that monolayers formed with the pure lipid have a liquid-like character; here, a hypothesis that admixing cholesterol to β-Mal(3)O(C(16+4))(2) could confer a higher rigidity on the films was tested. To check this proposal, two-dimensional miscibility of cholesterol and β-Mal(3)O(C(16+4))(2) in monomolecular films was studied using surface pressure and surface potential measurements, as well as Brewster angle microscopy and polarization-modulation infrared reflection absorption spectroscopy. The stability of the monomolecular films was evaluated based on thermodynamics of mixing of cholesterol and β-Mal(3)O(C(16+4))(2). Atomic level information concerning the orientation of molecules and the degree of hydration of polar headgroups was obtained from molecular dynamics simulations.     

Mitosis: towards a molecular understanding of chromosome behavior.

The past year has seen important contributions made to resolving how chromosomes attach to and move on the mitotic spindle of animal cells. These include the findings that: kinetochore microtubules are derived from the asters (i.e. centrosomes); poleward chromosome motion need not be coupled to kinetochore microtubules disassembly; the motor for poleward chromosome motion is associated with the kinetochore; and immunological evidence that this motor is cytoplasmic dynein.     

Metallomics and chemical speciation: towards a better understanding of metal-induced stress in plants

Most metal ions are toxic to plants, even at low concentrations, despite the fact that some are essential for growth and play key roles in metabolism. The majority of metals induce the formation of reactive oxygen species, which require the synthesis of additional antoxidant compounds and enzymes for their removal. New techniques that have greatly improved the identification, localisation and quantification of metals within plant tissues have led to the science of metallomics. This advancement in knowledge should eventually allow the characterisation of plants used in the process of phytoremediation of soils contaminated with toxic metals.     

The dilute locus and Griscelli syndrome: gateways towards a better understanding of melanosome transport

In this review an overview of recent advances in the understanding of melanosome movement within epidermal melanocytes is given. Exploration of the molecular events involved in and determining the process of melanosome transport, as an essential part of human pigmentation, could lead to the identification of agents that augment, or down-regulate the transfer of melanosomes to surrounding keratinocytes. This would present a major breakthrough in the possibilities to influence pigmentation and related disorders, of great concern to some patients. Moreover, melanosome transport offers a good model to study mammalian organelle trafficking and its key players in general.     

Nutritional sub-fertility in the dairy cow: towards improved reproductive management through a better biological understanding

There has been a significant decline in the reproductive performance of dairy cattle in recent decades. Cows, take longer time to return to the oestrus after calving, have poorer conception rates, and show fewer signs of oestrus. Achieving good reproductive performance is an increasing challenge for the dairy producer. In this study we focus on understanding the overall biological phenomena associated with nutritional sub-fertility rather than the underlying multiplicity of physiological interactions (already described in a number of recent studies). These phenomena are important because they represent the natural adaptations of the animal for dealing with variations in the nutritional environment. They can also be used to monitor and modulate reproductive performance on-farm. There is an underlying trade-off between two aspects of reproduction: investment in the viability of the current calf and investment in future offspring. As the investment in, and viability of, the current calf is related to maternal milk production, we can expect that level of milk production per se has effects on subsequent reproductive performance (investment in future offspring). Lactating cows have a lower proportion of viable embryos, which are of poorer quality, than do non-lactating cows. The same applies to high- compared to medium-genetic merit cows. Another important biological property is the adaptive use of body reserves in support of reproduction. Orchestrated endocrine changes in pregnancy and lactation facilitate the deposition of body lipid during pregnancy and mobilisation in early lactation. When the cow fails to accumulate the reserves she needs to safeguard reproduction she delays committing to further reproductive investment. But how does the cow ‘know’ that she is failing in energy terms? We argue that the cow does this by ‘monitoring’ both the body fat mobilisation and body fatness. Excessive body fat mobilisation indicates that current conditions are worse than expected. Body fatness indicates the future ability of the cow to safeguard her reproductive investment is compromised. Both delay further reproductive commitment. The relationship between reproductive performance and; milk production as an index of maternal investment, body fatness as an index of ability to safeguard reproductive investment, and body fat mobilisation as an index of the current nutritional environment – are examined. Nutritional strategies that seek to modulate body mobilisation and the endocrine environment by use of glucogenic and lipogenic diets, and the use of in-line progesterone profiles to monitor reproductive status are then discussed in this biological context.     

Perspectives on the therapeutic potential of short-chain fatty acid receptors

There is rapidly growing interest in the human microbiome because of its implication in metabolic disorders and inflammatory diseases. Consequently, understanding the biology of short chain fatty acids and their receptors has become very important for identifying novel therapeutic avenues. GPR41 and GPR43 have been recognized as the cognate receptors for SCFAs and their roles in metabolism and inflammation have drawn much attention in recent years. GPR43 is highly expressed on immune cells and has been suggested to play a role in inflammatory diseases such as inflammatory bowel disease. Both GPR41 and GPR43 have been implicated in diabetes and obesity via the regulation of adipose tissue and gastrointestinal hormones. So far, many studies have provided contradictory results, and therefore further research is required to validate these receptors as drug targets. We will also discuss the synthetic modulators of GPR41 and GPR43 that are critical to understanding the functions of these receptors. [BMB Reports 2014; 47(3): 173-178]     

How molecular modelling can better broaden the understanding of glycosylations

Glycosylations are among the most ubiquitous post-translational modifications (PTMs) in proteins, and the effects of their perturbations are seen in various diseases such as cancers, diabetes and arthritis to name a few. Yet they remain one of the most enigmatic aspects of protein structure and function. On the other hand, molecular modelling techniques have been rapidly bridging this knowledge gap since the last decade. In this review, we discuss how these techniques have proven to be indispensable for a better understanding of the role of glycosylations in glycoprotein structure and function.     

ER quality control: towards an understanding at the molecular level.

The process of 'quality control' in the endoplasmic reticulum (ER) involves a variety of mechanisms that collectively ensure that only correctly folded, assembled and modified proteins are transported along the secretory pathway. In contrast, non-native proteins are retained and eventually targeted for degradation. Recent work provides the first structural insights into the process of glycoprotein folding in the ER involving the lectin chaperones calnexin and calreticulin. Underlying principles governing the choice of chaperone system engaged by different proteins have also been discovered.     

New approaches to a better understanding of nematode phylogeny: molecular and developmental studies*

Recent studies making use of ribosomal DNA sequences have led to a wealth of new information that can be used to construct phylogenetic trees and compare them with those derived from more traditional approaches. Here, some examples are presented where the molecular data have stimulated alternative views. To determine whether differences in developmental processes can be used as markers for intraphyletic relationships, comparative studies of nematode embryogenesis have been performed. These results indicate that considerable deviations from the standard pattern found in the model system Caenorhabditis elegans exist which appear to be characteristic for certain subgroups within the taxon Nematoda.     

Molecular systematics and population genetics of biological invasions: towards a better understanding of invasive species management

The study of population genetics of invasive species offers opportunities to investigate rapid evolutionary processes at work, and while the ecology of biological invasions has enjoyed extensive attention in the past, the recentness of molecular techniques makes their application in invasion ecology a fairly new approach. Despite this, molecular biology has already proved powerful in inferring aspects not only relevant to the evolutionary biologist but also to those concerned with invasive species management. Here, we review the different molecular markers routinely used in such studies and their application(s) in addressing different questions in invasion ecology. We then review the current literature on molecular genetic studies aimed at improving management and the understanding of invasive species by resolving of taxonomic issues, elucidating geographical sources of invaders, detecting hybridisation and introgression, tracking dispersal and spread and assessing the importance of genetic diversity in invasion success. Finally, we make some suggestions for future research efforts in molecular ecology of biological invasions.     

The short-chain fatty acid butyrate is a substrate of breast cancer resistance protein

Colorectal cancer is one of the most common cancers worldwide. Butyrate (BT) plays a key role in colonic epithelium homeostasis. The aim of this work was to investigate the possibility of BT being transported by P-glycoprotein (MDR1), multidrug resistance proteins (MRPs), or breast cancer resistance protein (BCRP). Uptake and efflux of (14)C-BT and (3)H-folic acid were measured in Caco-2, IEC-6, and MDA-MB-231 cell lines. mRNA expression of BCRP was detected by RT-PCR. Cell viability, proliferation, and differentiation were quantified with the lactate dehydrogenase, sulforhodamine B, and alkaline phosphatase activity assays, respectively. In both IEC-6 cells and Caco-2 cells, no evidence was found for the involvement of either MDR1 or MRPs in (14)C-BT efflux from the cells. In contrast, several lines of evidence support the conclusion that BT is a substrate of both rat and human BCRP. Indeed, BCRP inhibitors reduced (14)C-BT efflux in IEC-6 cells, both BT and BCRP inhibitors significantly decreased the efflux of the known BCRP substrate (3)H-folic acid in IEC-6 cells, and BCRP inhibitors reduced (14)C-BT efflux in the BCRP-expressing MDA-MB-231 cell line. In IEC-6 cells, combination of BT with a BCRP inhibitor significantly potentiated the effect of BT on cell proliferation. The results of this study, showing for the first time that BT is a BCRP substrate, are very important in the context of the high levels of BCRP expression in the human colon and the anticarcinogenic and anti-inflammatory role of BT at that level. So, interaction of BT with BCRP and with other BCRP substrates/inhibitors is clearly of major importance.     

BUdR, probability and cell variants: towards a molecular understanding of the decision to differentiate

The mechanism(s) by which the thymidine analogue 5-bromodeoxyuridine (BUdR) specifically inhibits the expression of differentiated functions is poorly understood, as are the ways in which cells regulate processes exhibiting probabilistic aspects. I have developed a theoretical model for the regulation of the decision of myogenic cells to differentiate that can explain both of the above phenomena. This model provided a strategy for isolating myoblast variants that had amplified the expression of the factors regulating the decision to differentiate. These myoblasts served as the source of mRNA for making and screening a cDNA library in order to isolate these factors. The successful cloning of these genes should represent a major advance in our understanding of the molecular basis for the major coordinated changes in gene expression that accompany cell differentiation.     

Surface plasmon resonance: towards an understanding of the mechanisms of biological molecular recognition

With the introduction of new instruments and improved sensor chip chemistries, surface plasmon resonance (SPR) is finding new applications for molecular interaction studies. Easy access to high-quality kinetic and thermodynamic data for macromolecular binding events is providing insights into the fundamental mechanisms of molecular recognition. Progress is being made to allow larger-scale interaction studies. In addition, combining SPR with other analytical methods is enabling SPR-based analysis of interaction proteomics.     

Effect of short-chain fatty acids on the expression of genes involved in short-chain fatty acid transporters and inflammatory response in goat jejunum epithelial cells

Short-chain fatty acids (SCFAs) produced by microbial fermentation of dietary fibers are utilized by intestinal epithelial cells to provide an energy source for the ruminant. Although the regulation of mRNA expression and inflammatory response involved in SCFAs is established in other animals and tissues, the underlying mechanisms of the inflammatory response by SCFAs in goat jejunum epithelial cells (GJECs) have not been understood. Therefore, the objective of the study is to investigate the underlying mechanisms of the effects of SCFAs on SCFA transporters and inflammatory response in GJECs. These results showed that the acetate, butyrate, and SCFA concentration were markedly reduced in GJECs (p?<?0.01). In addition, the propionate concentration was significantly decreased in GJECs (p?<?0.05). The mRNA abundance of monocarboxylate transporter 1 (MCT1), MCT4, NHE1, and putative anion transporter 1 (PAT1) was elevated (p?<?0.05) by 20 mM SCFAs at pH 7.4 compared with exposure to the pH group. The anion exchanger 2 (AE2) was increased (p?<?0.05) by 20 mM SCFAs at pH 6.2. The mRNA abundance of vH⁺ ATPase B subunit (vH⁺ ATPase) was attenuated by SCFAs. For inflammatory responses, IL-1β and TNF-α were increased with SCFAs (p?<?0.05). In addition, IκBα involved in NF-κB signaling pathways was disrupted by SCFAs. Consistently, p-p65 signaling molecule was enhanced by adding SCFAs. However, IL-6 was attenuated by adding SCFAs (p?<?0.05). Furthermore, p-ErK1/2 mitogen-activated protein kinase (MAPK) signaling pathway was downregulated by adding SCFAs. In conclusion, these novel findings demonstrated that mRNA abundance involved in SCFA absorption is probably associated to SCFAs and pH value, and mechanism of the inflammatory response by SCFAs may be involved in NF-κB and p-ErK1/2 MAPK signaling pathways in GJECs. These pathways may mediate protective inflammation response in GJECs.     

Biosynthesis of triacylglycerols containing short-chain fatty acids in lactating cow mammary gland. Activity of diacylglycerol acyltransferase towards short-chain acyl-CoA esters

1. Microsomal 1,2-diacylglycerol acyltransferase from lactating cow mammary gland incorporated equal molar amounts of microsomal-bound 1,2-dipalmitoyl [2-3H]glycerol and [1-14C]-butyrate, [1-14C]hexanoate or [1-14C]palmitate from their CoA esters into triacylglycerol. The enzyme could also utilize exogenous 1,2-diacylglycerols in the presence of ethanol. 2. The pH optimum of the enzyme was 6.1 and 6.4 with butyryl-CoA and hexanoyl-CoA respectively. Values of V were approximately the same (2.7 and 2.4 nmol-min-1-mg-1, respectively), but values of Km were different (34 and 10 muM, respectively) with these two substrates. Mg2+ was not required as cofactor. 3. The presence ofa Mg2+-dependent phosphatidate phosphatase in the microsomal fraction was demonstrated. 4. It is proposed that triacylglycerols containing butyric and hexanoic acid are biosynthesized in cow mammary gland by the glycerolphosphate pathway, in which long-chain 1,2-diacylglycerols derived from phosphatidic acid are acylated at the sn-3 position by short-chain acyl-CoA esters.