Electrophoretic and kinetic studies of human erythrocytes deficient in pyrimidine 5′-nucleotidase

Summary A new case of a defect in red cell pyrimidine 5-nucleotidase (P5N) activity was found in a large family from Guadeloupe in the West Indies. The propositus presented a characteristic hemolytic anemia with red cell basophilic stippling, an increased GSH level, and a shift of the peak in absorbance of nucleotide. The enzyme activity of the deficient red cells was about 14% that of normal. The electrophoretic pattern of P5N activity from the deficient red cells differed from that of the normal. The P5N activity of the deficient red cells was distinct from that of the control in terms of its Km and of the effects of pH on its maximum activity and heat stability. The significance of such differences is discussed.     

Three-dimensional structure of the sugar symporter melibiose permease from cryo-electron microscopy

Melibiose permease (MelB) of Escherichia coli is a secondary transporter that couples the uptake of melibiose and various other galactosides to symport of cations that can be Na+, Li+ or H+. MelB belongs to the glycoside-pentoside-hexuronide: cation symporter family of porters and is suggested to have 12 transmembrane helices. We have determined the three-dimensional structure of MelB at 10A resolution in the membrane plane with cryo-electron microscopy from two-dimensional crystals. The three-dimensional map shows a heart-shaped molecule composed of two domains with a large central cavity between them. The structure is constricted at one side of the membrane while it is open to the other. The overall molecular shape resembles those of lactose permease and glycerol-3-phosphate transporter. However, organization of helices in MelB seems less symmetrical than in these two members of the major facilitator superfamily.     

Axonal transport of the cellular prion protein is increased during axon regeneration

The cellular prion protein, PrPc, is a glycosylphosphatidylinositol-anchored cell surface glycoprotein and a protease-resistant conformer of the protein may be the infectious agent in transmissible spongiform encephalopathies. PrPc is localized on growing axons in vitro and along fibre bundles that contain elongating axons in developing and adult brain. To determine whether the growth state of axons influenced the expression and axonal transport of PrPc, we examined changes in the protein following post-traumatic regeneration in the hamster sciatic nerve. Our results show (1) that PrPc in nerve is significantly increased during nerve regeneration; (2) that this increase involves an increase in axonally transported PrPc; and (3) that the PrPc preferentially targeted for the newly formed portions of the regenerating axons consists of higher molecular weight glycoforms. These results raise the possibility that PrPc may play a role in the growth of axons in vivo, perhaps as an adhesion molecule interacting with the extracellular environment through specialized glycosylation.     

Regulation of flower organogenesis: Phytohormone control of mRNA populations during sexual differentiation inMercurialis annua L.

Some general data on the genetic control and the possibilities of regulation of developmental paths inDrosophila are furnished. The insights to be gained from this insect will surely have implications that extend far beyond the fruit-fly. For example, in plants, developmental programs for floral organs, implying specific proteins are known. Developmental mutants in which mutate alleles control developmental programs for flowering were also selected in several species (Zea, Pisum, Sorghum, Cucumis, Mercurialis). Chemicals, especially phytohormones interfering with these programs are discussed. The case of sexual differentiation ofMercurialis is discussed in more detail. In this species, sex organs are controlled by sex determination genes and by auxins (male) and cytokinins (female). Flowers of each sex can be characterized by specific mRNA populations. They were evidenced by translationin vitro in a cell-free system of the various kinds of mRNAs [poly(A), non poly(A), polysomes]. The feminisation of genetic males by cytokinins induces the mRNA population of female type. Evidence concerning the implications of cytokinins in protein synthesis before translation level is presented. This is also probably true for auxins, although direct evidence is lacking.     

A novel symmetrical pyrano-3-deoxyanthocyanidin from a Sorghum species

Polyphenols are widespread components of higher plants. Sorghum is rich in polyphenols and known to contain a specific type, 3-deoxyanthocyanins, many of which remain uncharacterised. Here we report the structural determination of a new pigment isolated from red Sorghum bicolor var. bicolor (Moench) and characterised as 8-hydroxy-2-(4′-hydroxyphenyl)-5-(4″-hydroxyphenyl)-pyrano[4,3,2-de]1-benzopyrylium by MS, UV–vis and 2D NMR spectroscopy. This new symmetrical pyrano-3-deoxyanthocyanidin, containing apigeninidin as a base unit, displays structural features responsible for higher stability as compared to corresponding anthocyanins.     

Truncated PrP(c) in mammalian brain: interspecies variation and location in membrane rafts

A key molecular event in prion diseases is the conversion of cellular prion protein (PrP(c)) into an abnormal misfolded conformer (PrP(sc)). The PrP(c) N-terminal domain plays a central role in PrP(c) functions and in prion propagation. Because mammalian PrP(c) is found as a full-length and N-terminally truncated form, we examined the presence and amount of PrP(c) C-terminal fragment in the brain of different species. We found important variations between primates and rodents. In addition, our data show that the PrP(c) fragment is present in detergent-resistant raft domains, a membrane domain of critical importance for PrP(c) functions and its conversion into PrP(sc).     

Vegetation changes in Empakaai Crater, northern Tanzania, at 14,800–9300 cal yr BP

Vegetation changes are documented from a well-dated pollen record from Lake Emakat, Empakaai Crater, northern Tanzania. This pollen record includes the time interval covering the Pleistocene/Holocene transition, analysed at a resolution interval averaging 200 yr. Around the crater lake, an Hagenia-forest development starting at 14,500 cal yr BP lasted until 13,000 cal yr BP. A change in vegetation, indicated by an increased proportion of Nuxia congesta in the forest and Artemisia in the afro alpine grassland after 13,000 cal yr BP, corresponds in time to the Northern Hemisphere's Younger Dryas cooling. Grasses and sedges increased at  10,100 cal yr BP, indicating a significant increase in local pollen possibly attributed to lowered lake level, related to drier conditions. Although the Empakaai pollen record documents continuous forest conditions, from 14,500 to 10,100 cal yr BP, the variation in the proportion of forest components seem to respond to environmental changes at the millennium scale.     

Synergistic effect of hyaluronate fragments in retinaldehyde-induced skin hyperplasia which is a Cd44-dependent phenomenon

Background

CD44 is a polymorphic proteoglycan and functions as the principal cell-surface receptor for hyaluronate (HA). Heparin-binding epidermal growth factor (HB-EGF) activation of keratinocyte erbB receptors has been proposed to mediate retinoid-induced epidermal hyperplasia. We have recently shown that intermediate size HA fragments (HAFi) reverse skin atrophy by a CD44-dependent mechanism.

Methodology and Principal Findings

Treatment of primary mouse keratinocyte cultures with retinaldehyde (RAL) resulted in the most significant increase in keratinocyte proliferation when compared with other retinoids, retinoic acid, retinol or retinoyl palmitate. RAL and HAFi showed a more significant increase in keratinocyte proliferation than RAL or HAFi alone. No proliferation with RAL was observed in CD44^−/− keratinocytes. HA synthesis inhibitor, 4-methylumbelliferone inhibited the proliferative effect of RAL. HB-EGF, erbB1, and tissue inhibitor of MMP-3 blocking antibodies abrogated the RAL- or RAL- and HAFi-induced keratinocyte proliferation. Topical application of RAL or RAL and HAFi for 3 days caused a significant epidermal hyperplasia in the back skin of wild-type mice but not in CD44^−/− mice. Topical RAL and HAFi increased epidermal CD44 expression, and the epidermal and dermal HA. RAL induced the expression of active HB-EGF and erbB1. However, treatment with RAL and HAFi showed a more significant increase in pro-HB-EGF when compared to RAL or HAFi treatments alone. We then topically applied RAL and HAFi twice a day to the forearm skin of elderly dermatoporosis patients. After 1 month of treatment, we observed a significant clinical improvement.

Conclusions and Significance

Our results indicate that (i) RAL-induced in vitro and in vivo keratinocyte proliferation is a CD44-dependent phenomenon and requires the presence of HA, HB-EGF, erbB1 and MMPs, (ii) RAL and HAFi show a synergy in vitro and in vivo in mouse skin, and (iii) the combination of RAL and HAFi seems to have an important therapeutic effect in dermatoporosis.