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The abundance and size of cellular organelles vary depending on the cell type and metabolic needs. Peroxisomes constitute a class of cellular organelles renowned for their ability to adapt to cellular and environmental conditions. Together with transcriptional regulators, two groups of peroxisomal proteins have a pronounced influence on peroxisome size and abundance. Pex11-type peroxisome proliferators are involved in the proliferation of peroxisomes, defined here as an increase in size and/or number of peroxisomes. Dynamin-related proteins have recently been suggested to be required for the scission of peroxisomal membranes. This review surveys the function of Pex11-type peroxisome proliferators and dynamin-related proteins in peroxisomal proliferation and division.

Purpose: To examine the effect of apolipoprotein E (APOE) alleles on age-related macular degeneration (AMD) risk and on age at diagnosis of AMD in a large patient cohort recruited from a single center., Methods: The frequency of APOE alleles was analyzed in 632 unrelated AMD patients and 206 unrelated controls, all of whom were of white ancestry. The presence or absence of disease symptoms in all patients and controls was based on clinical examination and/or ophthalmic records. The association with APOE was explored in the context of AMD subtypes, family history status, possible interaction with smoking, and distribution of age at diagnosis of AMD., Results: The frequency of the epsilon4 allele was significantly reduced in patients compared with controls (0.10 vs. 0.14, P < or = 0.02). Gender- and age-adjusted odds ratios indicated that epsilon4-carriers have significantly lower risk of developing AMD compared to epsilon3epsilon3 subjects (OR = 0.55, 95% CI: 0.37-0.82, P = 0.004). In the cohort, AMD patients with a positive family history exhibited a significant 3.5 years earlier age at diagnosis (P = 0.001); however, APOE alleles did not appear to modulate the age at diagnosis of AMD., Conclusions: The association between the APOE-epsilon4 allele and a reduced risk of AMD was established in a large cohort with sufficient statistical power. How distinct APOE alleles affect AMD susceptibility warrants further investigation.

The formation and sinking of biogenic particles mediate vertical mass fluxes and drive elemental cycling in the ocean. Whereas marine sciences have focused primarily on particle production by phytoplankton growth, particle formation by the assembly of organic macromolecules has almost been neglected. Here we show, by means of a combined experimental and modelling study, that the formation of polysaccharide particles is an important pathway to convert dissolved into particulate organic carbon during phytoplankton blooms, and can be described in terms of aggregation kinetics. Our findings suggest that aggregation processes in the ocean cascade from the molecular scale up to the size of fast-settling particles, and give new insights into the cycling and export of biogeochemical key elements such as carbon, iron and thorium.

A new model for catalysis of human carbonic anhydrase II is suggested. The model is based on the X-ray structure of the hydrogen bond network in the catalytic site. The outer part of the network is proposed to adjust the p K(a) of the catalytic site to the experimentally observed value of about 7. The inner part of the network is proposed to become a low-barrier hydrogen bond network in the transition state. The energy released in forming the low-barrier hydrogen bond network is used to catalyse the interconversion of CO(2) and HCO(3)(-). The suggested molecular mechanism is consistent with the generally accepted kinetic scheme for human carbonic anhydrase II., (Copyright 2002 Elsevier Science Ltd. All rights reserved.)

The OLE pathway of yeast regulates the abundance of the ER-bound enzyme Delta-9 fatty acid desaturase OLE1, thereby controlling unsaturated fatty acid pools and membrane fluidity. Previously, we showed that this pathway is exquisitely regulated by the ubiquitin/proteasome system. Activation of the pathway involves proteasomal processing of a membrane-bound transcription factor and the subsequent mobilization of the cleaved, ubiquitylated transcription factor from its partner molecule by CDC48(UFD1/NPL4), a ubiquitin-selective chaperone-like enzyme. Here we report that the OLE1 protein itself is naturally short-lived and is degraded by ubiquitin/proteasome-dependent ER-associated degradation (ERAD). We found that CDC48(UFD1/NPL4) plays a second role in the OLE pathway by mediating ERAD of OLE1. Intriguingly, other ERAD substrates also require CDC48(UFD1/NPL4) for degradation, indicating that this enzyme is a novel, constitutive component of the ERAD machinery. We propose that CDC48(UFD1/NPL4) functions as a segregase that liberates ubiquitylated proteins from non-modified partners.

A generic chloroplast-based model for the carbon concentrating mechanism (CCM) in eukaryotic algae is presented. The fine structure of chloroplasts is represented by separate compartments: marginal and bulk stroma, pyrenoid, girdle lamella, bulk thylakoids, and central lamella traversing the pyrenoid. The roles of the individual structural elements of the chloroplast with respect to the CCM and the effect of carbonic anhydrase activity in various compartments are analysed. Hypothetical HCO(-)(3)transport into the acidic thylakoid lumen is adjusted by imposing an optimization principle: a given [CO(2)] at the site of RuBisCO is achieved with minimum energy costs for the CCM. Our model is highly efficient in terms of saturation of RuBisCO carboxylase activity and the affinity of the chloroplast for CO(2), if either a girdle lamella or a pyrenoid is present. The highest efficiency is achieved with a pyrenoid. A eukaryotic CCM is not necessarily associated with accumulation of dissolved inorganic carbon (DIC) as in cyanobacteria. Chloroplasts are categorized into four types corresponding to morphological characteristics of all major algal classes with regard to the presence of pyrenoids, girdle lamellae, and the distribution of CA activity., (Copyright 2001 Academic Press.)

A 28-year old woman with ovarian choriocarcinoma developed a uveitis in her left eye. Subsequently, a mass was observed in the anterior segment of that eye together with a subconjunctival mass that appeared to extend from it. Biopsy of the subconjunctival nodule showed two different cell types consistent with the cytotrophoblastic and syncytiotrophoblastic elements typical and choriocarcinoma. No abnormalities of the posterior segment were found on careful examination. Treatment consisted of systemic chemotherapy with a variety of therapeutic agents, argon laser photocoagulation of tumor seedings in the anterior chamber angle, radiation to the eye, and finally, subconjunctival injections of methotrexate. The eye became blind and painful and was enucleated. Histopathologic examination revealed residual tumor cells in the anterior segment indicating treatment changes, but there were no abnormalities posteriorly. Choriocarcinoma metastatic to the eye has been reported infrequently, and this is the first case in which anterior segment metastases have been observed and the effects of treatment thoroughly documented.

The effects of retinoic acid on the regeneration of double anterior lower arms in the adult newt, Notophthalmus viridescens, were investigated. Normally, double anterior lower arms regenerate a hypomorphic, symmetrical pattern of structures, which are distally complete; and double anterior upper arms regenerate a hypomorphic, symmetrical but distally incomplete pattern of structures. In limbs with a normal anteroposterior axis, the major effect of retinoic acid is to alter the proximodistal (PD) positional value of cells local at the amputation level to a much more proximal value, thereby creating duplications in the regenerate of structures proximal to the amputation plane (Thoms and Stocum, '84). Therefore, we predicted that double anterior lower arms treated with retinoic acid would regenerate like double anterior upper arms. However, in a substantial number of cases, each half of these double anterior lower arms regenerated a limb that was complete in the anteroposterior (AP) axis, with asymmetry corresponding to the half of origin. In addition, these regenerates were serially duplicated in the PD axis. These results indicate that retinoic acid can posteriorize the positional value of midline cells, leading to restoration of normal AP pattern, when the set of posterior-half positional values is removed from the cross section of the limb.

Pattern regulation following axial misalignments in the stage-38+to stage-40 urodele limb bud was studied on one newt and two salamander species. Grafts of the distal tip of the limb bud were made to the stump of a host limb bud from which a similar piece had been removed. The grafts were positioned with either their anteroposterior, dorsoventral, or both of these axes reversed with respect to the host axes. Mirror-imaged duplications, positioned posteriorly or both anteriorly and posteriorly, occurred nearly all (96%) of the time when the anteroposterior axis was reversed. Dorsoventral axial misalignment rarely promoted the generation of mirror-imaged duplication (8%) but did affect the organization along the anteroposterior axis by causing a serial repetition of either digit 2 or digit 3. Regulation, therefore, does not always occur along each axis independently of the others. Consistent with the data derived from reversing individual axes, most of the duplications which occurred when both axes were reversed were in the anteroposterior plane. Some were in the dorsoventral plane, and a few had intermediate positions. Of these duplications a few were neither right not left hands, rather they were of mixed handedness with a change in the dorsoventral polarity from the anterior border to the posterior border. Whether extra parts which result from axial misalignments arise from the graft, the host, or both the graft and the host was investigated using heteroplastic grafts and grafts exchanged between triploid and diploid axolotls. Duplications were found to have cellular contributions from both the graft and the host. In some cases on source would dominate but usually both made a substantial contribution. The diploid-triploid material suggests that a considerable mixing of host and graft cells may occur in duplications. Additionally, some digits of the graft sequence of digits can be derived from host tissue. The extra digit in those hands displaying a serial repetition was derived from host tissue in some cases and graft tissue in other cases.

The effects of varying doses of retinoic acid on forelimb regeneration in larval Ambystoma mexicanum amputated through the wrist joint and in adult Notophthalmus viridescens amputated through the basal carpals were compared. In both species, the major effect of retinoic acid was to cause the proximodistal duplication, in the regenerate, of stump segments proximal to the amputation plane. Transverse axial duplications (anteroposterior and dorsoventral) occurred in a smaller percentage of cases; these consisted of cartilage spurs in axolotls, and extra digits in newts. The frequency and magnitude of the proximodistal and (in the newt) transverse duplications were dose dependent, and the regenerating limbs were maximally sensitive to the retinoid during the period of dedifferentiation and accumulation of blastema cells. The effect of retinoic acid is exerted on cells local to the amputation surface, as shown by the fact that retinoic acid caused the proximodistal duplication of stump segments in regenerates derived from amputated distal lower arm segments grafted to the eyesocket .

The effects of two vitamin A analogues, retinol palmitate and retinoic acid, on pattern formation during limb regeneration in larvae of two European newts, P. waltl and T. vulgaris are described. The response of the regenerating limb to retinoid treatment differed according to the larval stage of development for P. waltl. Young larval limbs, which were anterior limb buds at the time of amputation, duplicated transversely while limbs of older larvae duplicated proximodistally. Proximodistal duplications were usually limited to the production of supernumerary carpals or a second zeugopod. Complete limbs regenerating from a distal amputation plane were rarely seen. T. vulgaris larvae regenerated limbs with either one or the other type of duplication, but never both on the same limb, at all larval stages tested. When larval P. waltl were kept in normal laboratory light during the treatment with retinol palmitate suspended in the rearing water the percentage of limbs which duplicated was very small for young larvae and increased with the age of the larvae used. Keeping the animals in the dark during the treatment period greatly increased the percentage of duplicate limbs obtained on the young larvae but not on the older larvae. This result is discussed in terms of the photodegradation of the retinoid and the length of the sensitive period for the regenerating limb. A dose-response relationship between the dose of retinol palmitate and either the percentage of limbs duplicated or the degree of duplication was not found. Such a relationship, however, was observed when retinoic acid was injected intraperitoneally into stage-54+ P. waltl larvae. Additionally, this technique revealed a peak of sensitivity to retinoic acid on the 6th day after amputation. Limb regeneration in older larvae was temporarily blocked by retinoid treatment. The limbs showed little or no regression and began blastemal development shortly after the treatment ended. Limbs of young larvae, however, often regressed. Such regressions were followed by blastemal formation and middle- to late-bud blastemas were found at the end of 11 or 14 days treatments with retinol palmitate.

Cyclic AMP (cAMP) appears to play multiple roles in the development of the social ameba Dictyostelium discoideum, serving as the chemoattractant mediating aggregation, and perhaps also regulating gene transcription in both early and late stages of differentiation. Progress in understanding the mechanism of activation of the adenylate cyclase in D. discoideum has been frustrated by the inability to obtain its activation in vitro. Also, the lack of defined cAMP-defective mutants has prevented a causal relationship from being established between cAMP levels and gene expression. As an alternative approach to studying the role of cAMP in D. discoideum development, we have sought a compound which inhibits cAMP synthesis in a reasonably specific manner. Here we identify caffeine as a compound which rapidly and reversibly inhibits cAMP-dependent activation of the adenylate cyclase without affecting either cell viability or intracellular levels of ATP or GTP. Using this drug, we show that cAMP synthesis is not required for the cAMP-stimulated decrease in lightscattering, the increase in cyclic GMP synthesis, or for chemotaxis toward cAMP. Studies of the mechanism of action of caffeine show that the drug does not act by inhibiting a cAMP phosphodiesterase, by inhibiting binding of cAMP to its receptor, by itself binding to a physiological adenosine receptor, or by directly inhibiting the adenylate cyclase. Instead, caffeine blocks the cAMP-dependent activation of the adenylate cyclase. Since similar effects are obtained with the cation ionophore A23187, it is possible that caffeine exerts its effect by altering intracellular calcium distribution.