El recorrido: evaluación de las cualidades sensoriales y formales del medio urbano como parte de los criterios de caminabilidad. La ciudad de Taipei como caso de estudio

La presente investigación aborda el análisis de las cualidades del entorno urbano que inciden en la experiencia perceptiva de su recorrido. Los intereses que la motivan tienen que ver con la recuperación de la ciudad para el viandante y se sitúa dentro de un marco de intereses más amplio como es el de la ciudad sostenible. En primer lugar, se contextualiza el tema de la experiencia del recorrido a través de disciplinas diferentes como son el cine, la literatura, la arquitectura, el urbanismo, la arquitectura del paisaje, la psicología y la estética ambiental. De este modo, se refuerza el argumento en torno al papel que desempeñan la forma y otros estímulos sensoriales en la experiencia del espacio, ya sea éste urbano, arquitectónico o natural. La experiencia que las personas viven al desplazarse por un entorno viene definida por infinitos factores que van desde la configuración física del entorno recorrido, hasta el aprendizaje cultural de la persona que camina; desde los estímulos sensoriales que se reciben, hasta las condiciones físicas que posibilitan la funcionalidad del recorrido; desde las sutilezas de índole secuencial que se desvelan a medida que avanzamos, hasta las condiciones higrotérmicas en el momento en que se lleva a cabo el desplazamiento. Sin embargo, factores como las motivaciones y atenciones personales del que anda, así como sus recuerdos de experiencias anteriores, deseos, imaginación y estado de ánimo, también definen nuestra experiencia al caminar que, indudablemente, se encuentra ligada a nuestra personalidad y, por lo tanto, no puede ser exacta a la de otra persona. Conscientes de que todos estos factores se funden en un fenómeno global, y que no se dan en la vida por separado, nos hemos esforzado por independizar las cualidades sensoriales y formales, únicamente a efectos de investigación. La importancia del tema a investigar reside en las conexiones que existen entre las cualidades del medio construido y la experiencia cotidiana de su recorrido, en la medida en que dicha vinculación establece relaciones con la calidad de vida, la salud de los ciudadanos y sus sentimientos de identidad. En este sentido, el problema clave encontrado es que si la definición de la experiencia del recorrido urbano es demasiado amplia, se vuelve nebulosa e inoperativa, pero si se precisa demasiado, se pueden excluir variables importantes. Hemos concentrado nuestros esfuerzos en buscar un medio que permita hacer operativo el estudio de aspectos que, por su propia naturaleza, son difíciles de controlar y aplicar en la práctica. En este sentido, el campo de estudio que aúna interés en el tema, producción científica y vocación práctica, es el de la caminabilidad de las ciudades. Por esta razón, hemos situado nuestro trabajo dentro del marco de criterios que este campo establece para valorar un entorno caminable. Sin embargo, hemos detectado que los trabajos sobre caminabilidad tienden a desarrollarse dentro de la disciplina del planeamiento de transporte y, con frecuencia, siguiendo las mismas pautas que la investigación sobre el transporte motorizado. Además, la tendencia a proporcionar datos numéricos producto de la medición controlada de variables, para respaldar iniciativas dentro de los ámbitos de toma de decisiones, lleva consigo un progresivo alejamiento de los aspectos más sutiles y próximos de la experiencia de las personas y la especificidad de los lugares. Por estas razones, hemos estimado necesario profundizar en la experiencia peatonal explorando otras líneas de trabajo como son la habitabilidad de las ciudades, políticas llevadas a cabo para mejorar la calidad del espacio público o las certificaciones de sostenibilidad en el ámbito del urbanismo. También se han estudiado las aproximaciones gráficas a la representación y simulación de recorridos urbanos por su importancia en la comunicación y promoción de caminar. Para detectar las problemáticas implicadas en la definición de una herramienta que permita valorar, de manera operativa, la calidad de la experiencia del recorrido urbano, la metodología de valoración propuesta se ha basado en la combinación de distintos métodos y en la conjugación de tres aproximaciones: valoración por parte del investigador en calidad de experto, valoración realizada por el investigador en el papel de usuario y valoración por el ciudadano. El desarrollo de esta investigación se ha visto condicionado por la intención de aplicar lo estudiado en un caso práctico. El marco de criterios que los trabajos sobre caminabilidad de las ciudades establecen para que se dé un entorno caminable, se puede resumir en que concurran los siguientes factores: mezcla de usos, densidad de población y edificación relativamente altas, destinos públicos accesibles a pie, un alto grado de seguridad con respecto al tráfico y actos delictivos, alta funcionalidad (dimensiones, pendientes, etc.) y atractivo. Una vez definido el modelo de ciudad en que es pertinente realizar una investigación sobre las cualidades del entorno urbano responsables de que éste se perciba como atractivo, se escogió la ciudad de Taipei, entre otras razones, por cumplir con los requisitos restantes. Con respecto al caso práctico, el objetivo es detectar fortalezas y debilidades del área central de la ciudad de Taipei en cuanto a la experiencia perceptiva que proporciona a los viandantes. ABSTRACT This research addresses the analysis of the qualities of the urban environment that affect the perceptual experience of walking. The interests lying behind it are related to the recovery of the city for pedestrians, and is framed within the sustainable city framework of interests. Firstly, the issue of the experience of walking is contextualized through different disciplines such as film, literature, architecture, urban planning, landscape architecture, environmental psychology and aesthetics. This way, the argument about the role that form and other sensory stimuli play in the experience of space, whether it is urban, architectural or natural, is strengthened. The walking experience of people is defined by factors ranging from the physical configuration of the environment to the cultural background of the person who walks, from the sensory stimuli that are perceived to the physical conditions that enable the functionality of the walk, from the subtleties of sequential nature that are revealed as we move around to hygrothermal conditions at the specific time of walking. Nevertheless, factors such as personal motivations and attentions of the walker, as well as memories of past experiences, desires, imagination and mood, also define our walking experience that is undoubtedly linked to our personality and, therefore, it cannot be exactly the same as other people's experience. Being aware that all these factors come together in a total phenomenon and that, in real life, they do not exist separately, we focused on separating sensory and formal qualities only for research purposes. The importance of the research topic lies in the connections between the qualities of the built environment and the quotidian experience of walking through it, to the extent that such link establishes relationships with the quality of life, health and feelings of identity of citizens. In this sense, the key problem encountered is that, if the definition of urban walking experience is too broad, it becomes nebulous and non‐operational, but if it is too precise, important variables can be excluded. We concentrated our efforts on finding a way to operationalize the study of questions that, by their very nature, are difficult to control and apply in practice. In this regard, the field of study that combines interest in the topic, scientific production and practical purpose, is the walkability of cities. For this reason, we placed our work within the framework of the set of criteria that this field establishes for assessing an environment as walkable. However, we found that works on walkability tend to be developed within the discipline of transportation planning and often following the same guidelines that research on motorized transport. Furthermore, the tendency to provide numerical data as a result of the controlled measurement of variables in order to support initiatives in decision making areas, involves a progressive distancing from the proximity and the most subtle aspects of the experience of people, and from the specificity of places. For these reasons, we considered it was necessary to deepen into the study of pedestrians experience, by exploring other lines of work such as the livability of cities, implemented policies to improve the quality of the public space or sustainability certifications in urbanism. Graphic representation and simulation of urban walks are also studied due to their important role in communication and promotion of walking. In order to find the issues involved in defining a tool to assess, in an operational way, the quality of urban walking experience, the proposed assessment methodology is based on the combination of different methods and the synthesis of three approaches: assessment by the researcher as expert, assessment by the researcher playing the role of user and assessment by the citizens. The development of this research has been conditioned by our intention of applying it in a case study. The framework of criteria that works on walkability of cities set to define a walkable environment, can be summarized in the following factors: mix of uses, population and building density rather high, public destinations accessible on foot, high levels of safety in terms of traffic and crime, high functionality (dimensions, slopes, etc.) and attractiveness. After defining the model of city in which it is relevant to conduct an investigation on the qualities that are responsible of the attractiveness of the environment; Taipei City was selected because it meets the remaining requirements, among other reasons. Regarding the case study, the goal is to identify strengths and weaknesses in the central area of Taipei City in terms of the perceptual experience of pedestrians.

Interaction of SP100 with HP1 proteins: A link between the promyelocytic leukemia-associated nuclear bodies and the chromatin compartment

The PML/SP100 nuclear bodies (NBs) were first described as discrete subnuclear structures containing the SP100 protein. Subsequently, they were shown to contain the PML protein which is part of the oncogenic PML-RARα hybrid produced by the t(15;17) chromosomal translocation characteristic of acute promyelocytic leukemia. Yet, the physiological role of these nuclear bodies remains unknown. Here, we show that SP100 binds to members of the heterochromatin protein 1 (HP1) families of non-histone chromosomal proteins. Further, we demonstrate that a naturally occurring splice variant of SP100, here called SP100-HMG, is a member of the high mobility group-1 (HMG-1) protein family and may thus possess DNA-binding potential. Both HP1 and SP100-HMG concentrate in the PML/SP100 NBs, and overexpression of SP100 leads to enhanced accumulation of endogenous HP1 in these structures. When bound to a promoter, SP100, SP100-HMG and HP1 behave as transcriptional repressors in transfected mammalian cells. These observations present molecular evidence for an association between the PML/SP100 NBs and the chromatin nuclear compartment. They support a model in which the NBs may play a role in certain aspects of chromatin dynamics.

A link density clustering algorithm based on automatically selecting density peaks for overlapping community detection

Peer reviewed

Thermus thermophilus: A link in evolution of the tRNA-dependent amino acid amidation pathways

Thermus thermophilus possesses an aspartyl-tRNA synthetase (AspRS2) able to aspartylate efficiently tRNAAsp and tRNAAsn. Aspartate mischarged on tRNAAsn then is converted into asparagine by an ω amidase that differs structurally from all known asparagine synthetases. However, aspartate is not misincorporated into proteins because the binding capacity of aminoacylated tRNAAsn to elongation factor Tu is only conferred by conversion of aspartate into asparagine. T. thermophilus additionally contains a second aspartyl-tRNA synthetase (AspRS1) able to aspartylate tRNAAsp and an asparaginyl-tRNA synthetase able to charge tRNAAsn with free asparagine, although the organism does not contain a tRNA-independent asparagine synthetase. In contrast to the duplicated pathway of tRNA asparaginylation, tRNA glutaminylation occurs in the thermophile via the usual pathway by using glutaminyl-tRNA synthetase and free glutamine synthesized by glutamine synthetase that is unique. T. thermophilus is able to ensure tRNA aminoacylation by alternative routes involving either the direct pathway or by conversion of amino acid mischarged on tRNA. These findings shed light on the interrelation between the tRNA-dependent and tRNA-independent pathways of amino acid amidation and on the processes involved in fidelity of the aminoacylation systems.

Link between light and fatty acid synthesis: Thioredoxin-linked reductive activation of plastidic acetyl-CoA carboxylase

Fatty acid synthesis in chloroplasts is regulated by light. The synthesis of malonyl-CoA, which is catalyzed by acetyl-CoA carboxylase (ACCase) and is the first committed step, is modulated by light/dark. Plants have ACCase in plastids and the cytosol. To determine the possible involvement of a redox cascade in light/dark modulation of ACCase, the effect of DTT, a known reductant of S-S bonds, was examined in vitro for the partially purified ACCase from pea plant. Only the plastidic ACCase was activated by DTT. This enzyme was activated in vitro more efficiently by reduced thioredoxin, which is a transducer of redox potential during illumination, than by DTT alone. Chloroplast thioredoxin-f activated the enzyme more efficiently than thioredoxin-m. The ACCase also was activated by thioredoxin reduced enzymatically with NADPH and NADP-thioredoxin reductase. These findings suggest that the reduction of ACCase is needed for activation of the enzyme, and a redox potential generated by photosynthesis is involved in its activation through thioredoxin as for enzymes of the reductive pentose phosphate cycle. The catalytic activity of ACCase was maximum at pH 8 and 2–5 mM Mg2+, indicating that light-produced changes in stromal pH and Mg2+ concentration modulate ACCase activity. These results suggest that light directly modulates a regulatory site of plastidic prokaryotic form of ACCase via a signal transduction pathway of a redox cascade and indirectly modulates its catalytic activity via stromal pH and Mg2+ concentration. A redox cascade is likely to link between light and fatty acid synthesis, resulting in coordination of fatty acid synthesis with photosynthesis.

A link between protein structure and enzyme catalyzed hydrogen tunneling

We present evidence that the size of an active site side chain may modulate the degree of hydrogen tunneling in an enzyme-catalyzed reaction. Primary and secondary kH/kT and kD/kT kinetic isotope effects have been measured for the oxidation of benzyl alcohol catalyzed by horse liver alcohol dehydrogenase at 25°C. As reported in earlier studies, the relationship between secondary kH/kT and kD/kT isotope effects provides a sensitive probe for deviations from classical behavior. In the present work, catalytic efficiency and the extent of hydrogen tunneling have been correlated for the alcohol dehydrogenase-catalyzed hydride transfer among a group of site-directed mutants at position 203. Val-203 interacts with the opposite face of the cofactor NAD+ from the alcohol substrate. The reduction in size of this residue is correlated with diminished tunneling and a two orders of magnitude decrease in catalytic efficiency. Comparison of the x-ray crystal structures of a ternary complex of a high-tunneling (Phe-93 → Trp) and a low-tunneling (Val-203 → Ala) mutant provides a structural basis for the observed effects, demonstrating an increase in the hydrogen transfer distance for the low-tunneling mutant. The Val-203 → Ala ternary complex crystal structure also shows a hyperclosed interdomain geometry relative to the wild-type and the Phe-93 → Trp mutant ternary complex structures. This demonstrates a flexibility in interdomain movement that could potentially narrow the distance between the donor and acceptor carbons in the native enzyme and may enhance the role of tunneling in the hydride transfer reaction.

The carbamate kinase-like carbamoyl phosphate synthetase of the hyperthermophilic archaeon Pyrococcus furiosus, a missing link in the evolution of carbamoyl phosphate biosynthesis

Microbial carbamoyl phosphate synthetases (CPS) use glutamine as nitrogen donor and are composed of two subunits (or domains), one exhibiting glutaminase activity, the other able to synthesize carbamoyl phosphate (CP) from bicarbonate, ATP, and ammonia. The pseudodimeric organization of this synthetase suggested that it has evolved by duplication of a smaller kinase, possibly a carbamate kinase (CK). In contrast to other prokaryotes the hyperthermophilic archaeon Pyrococcus furiosus was found to synthesize CP by using ammonia and not glutamine. We have purified the cognate enzyme and found it to be a dimer of two identical subunits of Mr 32,000. Its thermostability is considerable, 50% activity being retained after 1 h at 100°C or 3 h at 95°C. The corresponding gene was cloned by PCR and found to present about 50% amino acid identity with known CKs. The stoichiometry of the reaction (two ATP consumed per CP synthesized) and the ability of the enzyme to catalyze at high rate a bicarbonate-dependent ATPase reaction however clearly distinguish P. furiosus CPS from ordinary CKs. Thus the CPS of P. furiosus could represent a primeval step in the evolution of CPS from CK. Our results suggest that the first event in this evolution was the emergence of a primeval synthetase composed of subunits able to synthesize both carboxyphosphate and CP; this step would have preceded the duplication assumed to have generated the two subdomains of modern CPSs. The gene coding for this CK-like CPS was called cpkA.

Parkinson disease: A new link between monoamine oxidase and mitochondrial electron flow

Two factors that contribute to the progression of Parkinson disease are a brain defect in mitochondrial respiration and the generation of hydrogen peroxide (H2O2) by monoamine oxidase (MAO). Here we show that the two are linked. Metabolism of the neurotransmitter dopamine, or other monoamines (benzylamine, tyramine), by intact rat brain mitochondria suppresses pyruvate- and succinate-dependent electron transport. MAO inhibitors prevent this action. Mitochondrial damage is also reversed during electron flow. A probable explanation is that MAO-generated H2O2 oxidizes glutathione to glutathione disulfide (GSSG), which undergoes thiol-disulfide interchange to form protein mixed disulfides, thereby interfering reversibly with thiol-dependent enzymatic function. In agreement with this premise, direct addition of GSSG to mitochondria resulted in similar reversible inhibition of electron transport. In addition, the monoamines induced an elevation in protein mixed disulfides within mitochondria. These observations imply that (i) heightened activity and metabolism of neurotransmitter by monoamine neurons may affect neuronal function, and (ii) apparent defects in mitochondrial respiration associated with Parkinson disease may reflect, in part, an established increase in dopamine turnover. The experimental results also target mitochondrial repair mechanisms for further investigation and may, in time, lead to newer forms of therapy.

NF-κB activation provides the potential link between inflammation and hyperplasia in the arthritic joint

The transcription factor NF-κB is a pivotal regulator of inflammatory responses. While the activation of NF-κB in the arthritic joint has been associated with rheumatoid arthritis (RA), its significance is poorly understood. Here, we examine the role of NF-κB in animal models of RA. We demonstrate that in vitro, NF-κB controlled expression of numerous inflammatory molecules in synoviocytes and protected cells against tumor necrosis factor α (TNFα) and Fas ligand (FasL) cytotoxicity. Similar to that observed in human RA, NF-κB was found to be activated in the synovium of rats with streptococcal cell wall (SCW)-induced arthritis. In vivo suppression of NF-κB by either proteasomal inhibitors or intraarticular adenoviral gene transfer of super-repressor IκBα profoundly enhanced apoptosis in the synovium of rats with SCW- and pristane-induced arthritis. This indicated that the activation of NF-κB protected the cells in the synovium against apoptosis and thus provided the potential link between inflammation and hyperplasia. Intraarticular administration of NF-kB decoys prevented the recurrence of SCW arthritis in treated joints. Unexpectedly, the severity of arthritis also was inhibited significantly in the contralateral, untreated joints, indicating beneficial systemic effects of local suppression of NF-κB. These results establish a mechanism regulating apoptosis in the arthritic joint and indicate the feasibility of therapeutic approaches to RA based on the specific suppression of NF-κB.

Protein Interactions with the Glucose Transporter Binding Protein GLUT1CBP That Provide a Link between GLUT1 and the Cytoskeleton

Subcellular targeting and the activity of facilitative glucose transporters are likely to be regulated by interactions with cellular proteins. This report describes the identification and characterization of a protein, GLUT1 C-terminal binding protein (GLUT1CBP), that binds via a PDZ domain to the C terminus of GLUT1. The interaction requires the C-terminal four amino acids of GLUT1 and is isoform specific because GLUT1CBP does not interact with the C terminus of GLUT3 or GLUT4. Most rat tissues examined contain both GLUT1CBP and GLUT1 mRNA, whereas only small intestine lacked detectable GLUT1CBP protein. GLUT1CBP is also expressed in primary cultures of neurons and astrocytes, as well as in Chinese hamster ovary, 3T3-L1, Madin–Darby canine kidney, Caco-2, and pheochromocytoma-12 cell lines. GLUT1CBP is able to bind to native GLUT1 extracted from cell membranes, self-associate, or interact with the cytoskeletal proteins myosin VI, α-actinin-1, and the kinesin superfamily protein KIF-1B. The presence of a PDZ domain places GLUT1CBP among a growing family of structural and regulatory proteins, many of which are localized to areas of membrane specialization. This and its ability to interact with GLUT1 and cytoskeletal proteins implicate GLUT1CBP in cellular mechanisms for targeting GLUT1 to specific subcellular sites either by tethering the transporter to cytoskeletal motor proteins or by anchoring the transporter to the actin cytoskeleton.

Highly Stoichiometric, Stable, and Specific Association of Integrin α3β1 with CD151 Provides a Major Link to Phosphatidylinositol 4-Kinase, and May Regulate Cell Migration

Here we describe an association between α3β1 integrin and transmembrane-4 superfamily (TM4SF) protein CD151. This association is maintained in relatively stringent detergents and thus is remarkably stable in comparison with previously reported integrin–TM4SF protein associations. Also, the association is highly specific (i.e., observed in vitro in absence of any other cell surface proteins), and highly stoichiometric (nearly 90% of α3β1 associated with CD151). In addition, α3β1 and CD151 appeared in parallel on many cell lines and showed nearly identical skin staining patterns. Compared with other integrins, α3β1 exhibited a considerably higher level of associated phosphatidylinositol-4-kinase (PtdIns 4-kinase) activity, most of which was removed upon immunodepletion of CD151. Specificity for CD151 and PtdIns 4-kinase association resided in the extracellular domain of α3β1, thus establishing a novel paradigm for the specific recruitment of an intracellular signaling molecule. Finally, antibodies to either CD151 or α3β1 caused a ∼88–92% reduction in neutrophil motility in response to f-Met-Leu-Phe on fibronectin, suggesting an functionally important role of these complexes in cell migration.

Molecular basis for a link between complement and the vascular complications of diabetes

Activated terminal complement proteins C5b to C9 form the membrane attack complex (MAC) pore. Insertion of the MAC into endothelial cell membranes causes the release of growth factors that stimulate tissue growth and proliferation. The complement regulatory membrane protein CD59 restricts MAC formation. Because increased cell proliferation characterizes the major chronic vascular complications of human diabetes and because increased glucose levels in diabetes cause protein glycation and impairment of protein function, we investigated whether glycation could inhibit CD59. Glycation-inactivation of CD59 would cause increased MAC deposition and MAC-stimulated cell proliferation. Here, we report that (i) human CD59 is glycated in vivo, (ii) glycated human CD59 loses its MAC-inhibitory function, and (iii) inactivation of CD59 increases MAC-induced growth factor release from endothelial cells. We demonstrate by site-directed mutagenesis that residues K41 and H44 form a preferential glycation motif in human CD59. The presence of this glycation motif in human CD59, but not in CD59 of other species, may help explain the distinct propensity of humans to develop vascular proliferative complications of diabetes.

Calmodulin activates nuclear protein import: A link between signal transduction and nuclear transport

In addition to the well-characterized GTP-dependent nuclear transport observed in permeabilized cells, we detected a mode of nuclear transport that was GTP-independent at elevated cytoplasmic calcium concentrations. Nuclear transport under these conditions was blocked by calmodulin inhibitors. Recombinant calmodulin restored ATP-dependent nuclear transport in the absence of cytosol. Calmodulin-dependent transport was inhibited by wheat germ agglutinin consistent with transport proceeding through nuclear pores. We propose that release of intracellular calcium stores upon cell activation inhibits GTP-dependent nuclear transport; the elevated cytosolic calcium then acts through calmodulin to stimulate the novel GTP-independent mode of import.

A regenerative link in the ionic fluxes through the weaver potassium channel underlies the pathophysiology of the mutation

The homozygous weaver mouse displays neuronal degeneration in several brain regions. Previous experiments in heterologous expression systems showed that the G protein-gated inward rectifier K+ channel (GIRK2) bearing the weaver pore-region GYG-to-SYG mutation (i) is not activated by Gβγ subunits, but instead shows constitutive activation, and (ii) is no longer a K+-selective channel but conducts Na+ as well. The present experiments on weaverGIRK2 (wvGIRK2) expressed in Xenopus oocytes show that the level of constitutive activation depends on intracellular Na+ concentration. In particular, manipulations that decrease intracellular Na+ produce a component of Na+-permeable current activated via a G protein pathway. Therefore, constitutive activation may not arise because the weaver mutation directly alters the gating transitions of the channel protein. Instead, there may be a regenerative cycle of Na+ influx through the wvGIRK2 channel, leading to additional Na+ activation. We also show that the wvGIRK2 channel is permeable to Ca2+, providing an additional mechanism for the degeneration that characterizes the weaver phenotype. We further demonstrate that the GIRK4 channel bearing the analogous weaver mutation has properties similar to those of the wvGIRK2 channel, providing a glimpse of the selective pressures that have maintained the GYG sequence in nearly all known K+ channels.

Cryptocyanin, a crustacean molting protein: Evolutionary link with arthropod hemocyanins and insect hexamerins

Cryptocyanin, a copper-free hexameric protein in crab (Cancer magister) hemolymph, has been characterized and the amino acid sequence has been deduced from its cDNA. It is markedly similar in sequence, size, and structure to hemocyanin, the copper-containing oxygen-transport protein found in many arthropods. Cryptocyanin does not bind oxygen, however, and lacks three of the six highly conserved copper-binding histidine residues of hemocyanin. Cryptocyanin has no phenoloxidase activity, although a phenoloxidase is present in the hemolymph. The concentration of cryptocyanin in the hemolymph is closely coordinated with the molt cycle and reaches levels higher than hemocyanin during premolt. Cryptocyanin resembles insect hexamerins in the lack of copper, molt cycle patterns of biosynthesis, and potential contributions to the new exoskeleton. Phylogenetic analysis of sequence similarities between cryptocyanin and other members of the hemocyanin gene family shows that cryptocyanin is closely associated with crustacean hemocyanins and suggests that cryptocyanin arose as a result of a hemocyanin gene duplication. The presence of both hemocyanin and cryptocyanin in one animal provides an example of how insect hexamerins might have evolved from hemocyanin. Our results suggest that multiple members of the hemocyanin gene family—hemocyanin, cryptocyanin, phenoloxidase, and hexamerins—may participate in two vital functions of molting animals, oxygen binding and molting. Cryptocyanin may provide important molecular data to further investigate evolutionary relationships among all molting animals.