Reproducibility of Seasonal Effects in Antibody Formation and Host Resistance in the Outbred Mouse

The purpose of this study was to assess the reproducibility of any seasonal effects in the outbred CD1 mouse of antibody production to sheep red blood cells (SRBCs) and host resistance to the bacterium Listeria monocytogenes. A marked seasonal effect on antibody production was seen when 5- to 6-week-old female CD1 mice were studied on a weekly basis for a period of 2 years. Maintained on a 12:12 h lightdark schedule, animals were held for 12 days prior to experiment to insure physical condition and acclimatization to the lighting regimen. Beginning at 4 h after lights on (HALO) for day 1 and 2 HALO thereafter, groups of mice were (a) not treated, (b) administered a vehicle (corn oil, 1 % methylcellulose, or distilled water) by oral gavage for 5 days, or (c) not treated, but given an intraperitoneal injection of cyclophosphamide 24 h prior to assay. On the fifth day, mice were injected with SRBCs intravenously. Four days later, antibody formation against SRBCs was measured using spleen cells. Circannual and seasonal rhythms were displayed by each group of animals, with greatest antibody production, indicated by the number of plaque-forming cells (PFCs)/million viable cells, in the Spring (range of double amplitude = 36-108%). The timing of these rhythms was reproducible from one year to the next. In contrast, the magnitude of the response in year 1 was significantly different from year 2 for animals given vehicle or not treated. Cyclophosphamide-treated mice had consistently low numbers of plaque-forming cells. Host resistance was studied in separate mice treated with vehicles at the same time as the antibody assay. On the third day of dosing, mice were injected intravenously with Listeria monocytogenes (LM) and monitored for death for 10 days. When analyzed by Kruskal-Wallis life table analysis, there was no overall effect of vehicle on survival for 1987 but there was an effect for 1988 and when data from both years were combined. Distilled water-treated mice had lower survival rates than the other two vehicles. Mice treated with distilled water displayed a circannual rhythm for survival for 1988 and for both years combined, in contrast to the other two vehicles. When each vehicle was analyzed separately for seasonal effect, a significant effect of season occurred for corn oil- and distilled water-treated animals. The greatest survival rate and longest survival time generally occurred in the months between July and December.     

Evidence for Coexistence of Distinct Escherichia coli Populations in Various Aquatic Environments and Their Survival in Estuary Water

Escherichia coli, a commensal bacterium from the intestinal tracts of humans and vertebrate animals, has been used as one of two bacterial indicators of fecal contamination, along with intestinal enterococci, to monitor the microbiological quality of water. However, water environments are now recognized as a secondary habitat where some strains can survive. We investigated the survival of E. coli isolates collected from bodies of water in France exhibiting distinct profiles of contamination, defined according to the following criteria: vicinity of the point sources of contamination, land use, hydrology, and physicochemical characteristics of the receiving water. We selected 88 E. coli strains among a collection of 352 strains to carry out a microcosm experiment in filtered estuarine water for 14 days at 10°C. The relationship between the survival of E. coli strains and genotypic and phenotypic characteristics was analyzed. This work showed that distinct E. coli survival types, able to survive from between 7 and 14 days to less than 2 days, coexisted in the water. E. coli isolates that rapidly lost their culturability were more frequently isolated in water recently contaminated by fecal bacteria of human origin, and most were multiresistant to antibiotics and harbored several virulence factors. In contrast, persistent strains able to survive from 4 to 14 days were more often found in water with low levels of fecal bacteria, belonged mainly to the B1 phylogroup, often harbored only one virulence factor, kspE or ompT, and were able to grow at 7°C.     

The role of sugar signaling in plant defense responses against fungal pathogens

In most fungal pathogen–plant systems, a high level of sugars in plant tissues enhances plant resistance. Several hypotheses have been proposed to explain the mechanisms of “high-sugar resistance”. Sugars constitute the primary substrate providing energy and structural material for defense responses in plants, while they may also act as signal molecules interacting with the hormonal signaling network regulating the plant immune system. Sugars enhance oxidative burst at early stages of infection, increasing lignification of cell walls, stimulate the synthesis of flavonoids and induce certain PR proteins. Some sugars act as priming agents inducing higher plant resistance to pathogens.     

Evidence that the population of quiescent bone marrow‐residing very small embryonic/epiblast‐like stem cells (VSELs) expands in response to neurotoxic treatment

The concept that bone marrow (BM)-derived cells may participate in neural regeneration remains controversial, and the identity of the specific cell type(s) involved remains unknown. We recently reported that the adult murine BM contains a highly mobile population of Sca-1⁺Lin⁻CD45⁻ cells known as very small embryonic/epiblast-like stem cells (VSELs) that express several markers of pluripotency such as Oct-4. In the BM microenvironment, these cells are kept quiescent because of epigenetic modification of certain paternally imprinted genes. However, as reported, these cells can be mobilized in mice in an experimental model of stroke and express several genes involved in neurogenesis while circulating in peripheral blood (PB). In the current work, we employed a model of toxic brain damage, which is induced by administration of kainic acid, to see not only whether VSELs can be mobilized into PB in response to this neurotoxin, but, more importantly, whether they proliferate and expand in BM tissue. We report here for the first time that brain damage leads to activation and expansion of the BM pool of quiescent VSELs, which precedes their subsequent egress into PB. Harnessing these cells in neural tissue regeneration is currently one of the challenges in regenerative medicine.     

Regulation of storage lipid metabolism in developing and germinating lupin (<Emphasis Type="Italic">Lupinus</Emphasis> spp.) seeds

The main storage compound in lupin seeds is protein, whose content can reach up to 45–50 % of dry matter. However, seeds of some lupin species can also contain quite a large amount of storage lipid. The range of lipid content in lupin seeds is from about 6 to about 20 % of dry matter. Storage lipid in developing seeds is synthesized mainly from sugars delivered by mother plants. During seed germination, one of the main end-products of storage lipid breakdown is also sugars. Thus, the sugar level in tissues is considered an important regulatory agent, during both lipid accumulation and lipid breakdown. Generally, in developing legume seeds, there is a strong negative relation between accumulation of storage protein and storage lipid. Results obtained in developing lupin cotyledons cultured in vitro pointed to the possibility of a positive relation between protein and lipid accumulation. Such a positive effect could be caused by nitrate. During lupin seed germination and seedling development, the utilization of storage lipid is enhanced under sugar deficiency conditions in tissues and is controlled at the gene expression level. However, under sugar starvation conditions, autophagy is significantly enhanced, and it can cause disturbances in storage lipid breakdown. The hypothesis of pexophagy, i.e., autophagic degradation of peroxisomes under sugar starvation conditions during lupin seed germination, has been taken into consideration. The flow of lipid-derived carbon skeletons to amino acids was discovered in germinating lupin seeds, and this process is clearly more intense in sucrose-fed embryo axes. At least four alternative or mutually complementary pathways of carbon flow from storage lipid to amino acids in germinating lupin seeds are postulated. The different strategies of storage compound breakdown during lupin seed germination are also discussed.     

Transplantation of expanded bone marrow‐derived very small embryonic‐like stem cells (VSEL‐SCs) improves left ventricular function and remodelling after myocardial infarction

Adult bone marrow‐derived very small embryonic‐like stem cells (VSEL‐SCs) exhibit a Sca‐1⁺/Lin^–/CD45^– phenotype and can differentiate into various cell types, including cardiomyocytes and endothelial cells. We have previously reported that transplantation of a small number (1 × 10⁶) of freshly isolated, non‐expanded VSEL‐SCs into infarcted mouse hearts resulted in improved left ventricular (LV) function and anatomy. Clinical translation, however, will require large numbers of cells. Because the frequency of VSEL‐SCs in the marrow is very low, we examined whether VSEL‐SCs can be expanded in culture without loss of therapeutic efficacy. Mice underwent a 30 min. coronary occlusion followed by reperfusion and, 48 hrs later, received an intramyocardial injection of vehicle (group I, n= 11), 1 × 10⁵ enhanced green fluorescent protein (EGFP)‐labelled expanded untreated VSEL‐SCs (group II, n= 7), or 1 × 10⁵ EGFP‐labelled expanded VSEL‐SCs pre‐incubated in a cardiogenic medium (group III, n= 8). At 35 days after myocardial infarction (MI), mice treated with pre‐incubated VSEL‐SCs exhibited better global and regional LV systolic function and less LV hypertrophy compared with vehicle‐treated controls. In contrast, transplantation of expanded but untreated VSEL‐SCs did not produce appreciable reparative benefits. Scattered EGFP⁺ cells expressing α‐sarcomeric actin, platelet endothelial cell adhesion molecule (PECAM)‐1, or von Willebrand factor were present in VSEL‐SC‐treated mice, but their numbers were very small. No tumour formation was observed. We conclude that VSEL‐SCs expanded in culture retain the ability to alleviate LV dysfunction and remodelling after a reperfused MI provided that they are exposed to a combination of cardiomyogenic growth factors and cytokines prior to transplantation. Counter intuitively, the mechanism whereby such pre‐incubation confers therapeutic efficacy does not involve differentiation into new cardiac cells. These results support the potential therapeutic utility of VSEL‐SCs for cardiac repair.     

Cannabinoid-like anti-inflammatory compounds from flax fiber

Flax is a valuable source of fibers, linseed and oil. The compounds of the latter two products have already been widely examined and have been proven to possess many health-beneficial properties. In the course of analysis of fibers extract from previously generated transgenic plants overproducing phenylpropanoids a new terpenoid compound was discovered.The UV spectra and the retention time in UPLC analysis of this new compound reveal similarity to a cannabinoid-like compound, probably cannabidiol (CBD). This was confirmed by finding two ions at m/z 174.1 and 231.2 in mass spectra analysis. Further confirmation of the nature of the compound was based on a biological activity assay. It was found that the compound affects the expression of genes involved in inflammatory processes in mouse and human fibroblasts and likely the CBD from Cannabis sativa activates the specific peripheral cannabinoid receptor 2 (CB2) gene expression. Besides fibers, the compound was also found in all other flax tissues. It should be pointed out that the industrial process of fabric production does not affect CBD activity.The presented data suggest for the first time that flax products can be a source of biologically active cannabinoid-like compounds that are able to influence the cell immunological response. These findings might open up many new applications for medical flax products, especially for the fabric as a material for wound dressing with anti-inflammatory properties.     

Adipose tissue as a potential source of hematopoietic stem/progenitor cells

It has been more than 30 years since adipose tissue (AT) has been recognized as a central modulator orchestrating sophisticated process termed "immunometabolism". Nonetheless, despite its unique involvement in the regulation of immune and endocrine homeostasis, recent studies demonstrated that AT also contains significant number of hematopoietic stem/progenitor cells (HSPCs) that may be there "settling down" throughout life. In this article we will focus on presenting the current concepts regarding endocrine, immunological, and molecular mechanisms that may contribute to and regulate bone marrow (BM)-derived HSPCs homing into AT environment, as well as, highlight various structural and morphological similarities between BM and AT that might be involved in creating appropriate tissue niches for BM-derived HSPCs in AT. Finally, we will discuss how development of obesity or type 2 diabetes may influence balance of homing signals for HSPCs in AT environment.     

Population of Rh123dim human keratinocytes form holoclones

The aim of our studies was to develop an efficient strategy to isolate human early epidermal progenitors for experimental and potential clinical purposes. We employed fluorescence-activated cell sorting (FACS) to isolate cells that poorly accumulate metabolic Rhodamin123 (Rh123) dye. We noticed that similarly to a population of β1-integrin bright (β1^bright) cells, a population of Rh123 dull (Rh123^dim) cells is highly enriched for cells growing holoclones, colonies composed of the most primitive keratinocytes. Furthermore, Rh123^dim cells express several morphological features of primitive undifferentiated cells and are also highly motile. We postulate that these cells could become an important source of epidermal progenitors to expand keratinocytes for clinical purposes.     

The structure of barley alpha-amylase isozyme 1 reveals a novel role of domain C in substrate recognition and binding: a pair of sugar tongs

Though the three-dimensional structures of barley alpha-amylase isozymes AMY1 and AMY2 are very similar, they differ remarkably from each other in their affinity for Ca(2+) and when interacting with substrate analogs. A surface site recognizing maltooligosaccharides, not earlier reported for other alpha-amylases and probably associated with the different activity of AMY1 and AMY2 toward starch granules, has been identified. It is located in the C-terminal part of the enzyme and, thus, highlights a potential role of domain C. In order to scrutinize the possible biological significance of this domain in alpha-amylases, a thorough comparison of their three-dimensional structures was conducted. An additional role for an earlier-identified starch granule binding surface site is proposed, and a new calcium ion is reported.