Regenerative Xylem in Inflorescence Stems of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

By inserting entomological needles into the lower parts of young inflorescence stems of three-month-old Arabidopsis thaliana (L.) Heynh var. Colombia plants, we studied the process of regenerative xylem production. Regenerative xylem was formed only in one- to two-day-old inflorescence stems but not in older ones. The regenerative vessels originated from re-differentiation of cortical parenchyma. To characterize the process of regenerative xylem formation, we conducted a histological study from the time of wounding to day 30 after wounding. In the first day after wounding the tissues showed no structural responses except for the wounding itself. After six days, regenerative vessel members were already differentiating in a basipetal pattern, forming a vascular bypass around the wound. Regenerative vessel member formation reached a maximal level on the twelfth day after wounding. Sixteen days after wounding the pith parenchyma started to become loose as if indicating tissue senescence. Altogether, vascular regeneration following wounding in inflorescence stems of Arabidopsis thaliana is similar to that in other dicotyledon plants. These findings provide the basis for the use of Arabidopsis thaliana as a model system to study the genetics, physiology and cell biology of wound healing and regenerative vascular tissue formation.     

Complexes of HLA-G protein on the cell surface are important for leukocyte Ig-like receptor-1 function

The nonclassical class I MHC molecule HLA-G is selectively expressed on extravillous cytotrophoblast cells at the maternal-fetal interface during pregnancy. HLA-G can inhibit the killing mediated by NK cells via interaction with the inhibitory NK cell receptor, leukocyte Ig-like receptor-1 (LIR-1). Comparison of the sequence of the HLA-G molecule to other class I MHC proteins revealed two unique cysteine residues located in positions 42 and 147. Mutating these cysteine residues resulted in a dramatic decrease in LIR-1 Ig binding. Accordingly, the mutated HLA-G transfectants were less effective in the inhibition of NK killing and RBL/LIR-1 induced serotonin release. Immunoprecipitation experiments demonstrated the involvement of the cysteine residues in the formation of HLA-G protein oligomers on the cell surface. The cysteine residue located at position 42 is shown to be critical for the expression of such complexes. These oligomers, unique among the class I MHC proteins, probably bind to LIR-1 with increased avidity, resulting in an enhanced inhibitory function of LIR-1 and an impaired killing function of NK cells.     

Effects of transgene expression of superoxide dismutase and glutathione peroxidase on pulmonary epithelial cell growth in hyperoxia

Prolonged exposure to supraphysiological oxygen concentrations results in the generation of reactive oxygen species, which can cause significant lung injury in critically ill patients. Supplementation with human recombinant antioxidant enzymes (AOE) may mitigate hyperoxic lung injury, but it is unclear which combination and concentration will optimally protect pulmonary epithelial cells. First, stable cell lines were generated in alveolar epithelial cells (MLE12) overexpressing one or more of the following AOE: Mn superoxide dismutase (MnSOD), CuZnSOD, or glutathione peroxidase 1. Next, A549 cells were transduced with 50-300 particles/cell of recombinant adenovirus containing either LacZ or each of the three AOE (alone or in combination). Cells were then exposed to 95% O(2) for up to 3 days, with cell number and viability determined daily. Overexpression of either MnSOD (primarily mitochondrial) or CuZnSOD (primarily cytosolic) reversed the growth inhibitory effects of hyperoxia within the first 48 h of exposure, resulting in a significant increase in viable cells (P < 0.05), with 1.5- to 3-fold increases in activity providing optimal protection. Protection from mitochondrial oxidation was confirmed by assessing aconitase activity, which was significantly improved in cells overexpressing MnSOD (P < 0.05). Data indicate that optimal protection from hyperoxic injury occurs in cells coexpressing MnSOD and glutathione peroxidase 1, with prevention of mitochondrial oxidation being a critical factor. This has important implications for clinical trials in preterm infants receiving SOD supplementation to prevent acute and chronic lung injury.     

Aposematic (warning) coloration associated with thorns in higher plants

Aposematic coloration, a well-known phenomenon in animals, has been given little attention in plants. Here I discuss two types of conspicuousness of thorns which are typical of many plant species: (1) colorful thorns, and (2) white spots, or white and colorful stripes, associated with thorns in leaves and stems. Both types of aposematic coloration predominate the spine system of taxa rich with spiny species-Cacti, the genera Agave, Aloe and Euphorbia. The phenomena have been recorded here in over a thousand species originating in several continents of both the Old and New World. I propose that this is a case of vegetal aposematic coloration analogous to such coloration of poisonous animals, and which communicates between plants and herbivores.     

Terminology for plant polarity: Progress and difficulties

The recent attempts to establish a “natural” terminology for plant polarity^1–3 are significant as they have identified various problems in the classic terminology of polarity,⁴ and arrived at a logical and simple terminology (shootward and rootward).³ The related issue of polarity changes was also addressed while assuming that cell polarity is conserved throughout the development.²Key words: auxin, axiality, development, differentiation, epiphylly, plant architectureWhile the shootward and rootward terminology is very appealing and seems to describe “normal” plant development precisely, like in the model plant Arabidopsis thaliana, there are many instances in vascular plants that only partly fit the suggested terminology. I demonstrate it by describing three types of common rootward-shootward polarity changes in mature Angiosperm plants (out of a longer list), which are not solved by the new terminology.

1. Epiphylly of plantlets with roots formed on the margins of young leaves in Bryophyllum and other taxa.⁵ These roots result in a mixed polarity. For instance, the lower stem parts that had for a while only one rootward polarity acquire two contrasting directions of rootward polarity following the formation of many plantlet roots on leaf margins. Similarly, for the original roots, the young shoot was initially purely shootward but become at least partly of rootward polarity because of the formation of new plantlet roots on the leaves. Thus, in such plants a situation of rootward and shootward polarities originating from the same direction is found.
2. Aerial roots of many trees, e.g., of many Ficus species.⁵ These roots, which sometimes become larger than the original root system, must influence polarity to a large extent as discussed in the above example of much smaller roots.
3. Shoots emerging from roots of Populus, Rubus and many other taxa.⁵ Such shoots, formed at the rootward position relative to the root parts positioned closer to the original shoot, greatly alter the polarity status of the plants. They result in shootward and rootward polarities arriving from the same direction. Since many such shoots may develop along the root system, they result in an increasing complication of a mosaic of rootward and shootward polarities.

For all three above cases there is no data about where these opposite polarities meet, how they interact, and if there are physiological or developmental consequences to their signal cross-talk.I conclude that while the new shootward and rootward terminology³ is perfect in many cases throughout a plant''s life cycle and for practically all young vascular plants. However, this terminology needs some modifications or additions to fit other developmental situation found in many thousands of species that naturally or even artificially form ectopic roots or shoots. I therefore propose to distinguish between primary, secondary, tertiary and so on, ranks of shootward and rootward polarities.     

Glucocorticosteroids in Nano-Sterically Stabilized Liposomes Are Efficacious for Elimination of the Acute Symptoms of Experimental Cerebral Malaria

Cerebral malaria is the most severe complication of Plasmodium falciparum infection, and a leading cause of death in children under the age of five in malaria-endemic areas. We report high therapeutic efficacy of a novel formulation of liposome-encapsulated water-soluble glucocorticoid prodrugs, and in particular β-methasone hemisuccinate (BMS), for treatment of experimental cerebral malaria (ECM), using the murine P. berghei ANKA model. BMS is a novel derivative of the potent steroid β-methasone, and was specially synthesized to enable remote loading into nano-sterically stabilized liposomes (nSSL), to form nSSL-BMS. The novel nano-drug, composed of nSSL remote loaded with BMS, dramatically improves drug efficacy and abolishes the high toxicity seen upon administration of free BMS. nSSL-BMS reduces ECM rates in a dose-dependent manner and creates a survival time-window, enabling administration of an antiplasmodial drug, such as artemisone. Administration of artemisone after treatment with the nSSL-BMS results in complete cure. Treatment with BMS leads to lower levels of cerebral inflammation, demonstrated by changes in cytokines, chemokines, and cell markers, as well as diminished hemorrhage and edema, correlating with reduced clinical score. Administration of the liposomal formulation results in accumulation of BMS in the brains of sick mice but not of healthy mice. This steroidal nano-drug effectively eliminates the adverse effects of the cerebral syndrome even when the treatment is started at late stages of disease, in which disruption of the blood-brain barrier has occurred and mice show clear signs of neurological impairment. Overall, sequential treatment with nSSL-BMS and artemisone may be an efficacious and well-tolerated therapy for prevention of CM, elimination of parasites, and prevention of long-term cognitive damage.     

ERβ Isoforms Have Differential Clinical Significance in Breast Cancer Subtypes and Subgroups

Simple SummaryERβ, an ER subtype first identified in 1996, is significantly expressed in ERα-negative breast cancer (BCa) and TNBC. Many studies investigated mostly ERβ1 protein expression in the entire cohort of BCa, and the results are inconsistent. In this study, we simultaneously investigated both ERβ mRNA and three ERβ 1, 2, and 5 protein isoforms in various subtypes and subgroups of BCa. Each ERβ isoform’s mRNA and protein expression seemingly plays a significant role in BCa subtypes and subgroups, and ERβ2 mRNA expression is risk factor for poor outcome. Studies in a large cohort of BCa are needed to explore the potential usefulness of ERβ as a prognostic and predictive marker and a therapeutic target in BCa. Furthermore, the standardization of a ERβ testing protocol may be required for ERβ testing to be utilized in a clinical setting.AbstractERβ, an ER subtype first identified in 1996, is highly expressed in different types of BCa including ERα-negative BCa and TNBC. Many studies on ERβ expression investigated mostly on ERβ1 protein expression in ERα-positive and ERα-negative BCa combined. The results are conflicting. This may be due to the complexity of ERβ isoforms, subject heterogeneity, and various study designs targeting different ERβ isoforms and either ERβ protein or mRNA expression, as well as to the lack of a standardized testing protocol. Herein, we simultaneously investigated both mRNA and protein expression of ERβ isoforms 1, 2, and 5 in different BCa subtypes and clinical characteristics. Patient samples (138) and breast cancer cell lines (BCC) reflecting different types of BCa were tested for ERα and ERβ mRNA expression using quantitative real-time PCR, as well as for protein expression of ERα, ERβ1, ERβ2, and ERβ5 isoforms, PR, HER2/neu, Ki-67, CK 5/6, and p53 using immunohistochemistry. Associations of ERβ isoform expression with clinical characteristics and overall survival (OS) were analyzed. ERβ1, 2, and 5 isoforms are differentially expressed in different BCa subtypes including ERα-negative and TNBC. Each ERβ isoform seemingly plays a distinct role and is associated with clinical tumor characteristics and patient outcomes. ERβ isoform expression is significantly associated with >15% Ki-67 positivity and poor prognostic markers, and it predicts poorer OS, mostly in the subgroups. High ERβ2 and 5 isoform expression in ERα-negative BCa and TNBC is predictive of poor OS. Further investigation of ERβ isoforms in a larger cohort of BCa subgroups is needed to evaluate the role of ERβ for the potential usefulness of ERβ as a prognostic and predictive marker and for therapeutic use. The inconsistent outcomes of ERβ isoform mRNA or protein expression in many studies suggest that the standardization of ERβ testing would facilitate the use of ERβ in a clinical setting.     

Endometrial NK cells are special immature cells that await pregnancy

NK cells populate the human endometrium before pregnancy. Unlike decidual NK cells that populate the decidua during pregnancy, the NK cells present in the human endometrium, before pregnancy, have not been fully characterized. In this study, we provide a detailed analysis of the origin, phenotype, and function of endometrial NK cells (eNK). We show that eNK cells have a unique receptor repertoire. In particular, they are negative for NKp30 and chemokine receptor expression, which distinguishes them from any other NK subset described so far. We further show that eNK cells lack NK-specific functional phenotype and activity such as cytokine secretion and cytotoxicity, before IL-15 stimulation. Following such stimulation, endometrial NK cells acquire phenotype and function that are similar to those of decidual NK cells. We therefore suggest that eNK cells are inactive cells (before IL-15 activation and in relation to the known NK activity) that are present in the endometrium before conception, waiting for pregnancy.