Epiescleritis y edema periorbitario secundario a síndrome VEXAS (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic) / Episcleritis and periorbital edema secondary to VEXAS syndrome

El síndrome vacuolas, E1 enzyme, X-linked, autoinflammatory, somatic (VEXAS) es una nueva entidad autoinflamatoria descrita recientemente, producida por una mutación del gen UBA-1. Entre los síntomas más frecuentes están la fiebre, las citopenias, la policondritis, los infiltrados pulmonares y hasta en un 40% afectación ocular en forma de edema periorbitario, uveítis, epiescleritis, escleritis y vasculitis retiniana. Los pacientes responden a altas dosis de corticoterapia, sin embargo muchos terminan siendo refractarios a las mismas y a los inmunosupresores clásicos. Se describe el caso de un paciente varón de 77 años con afectación ocular en forma de epiescleritis y edema periorbitario que posteriormente fue diagnosticado de síndrome VEXAS. El paciente, tras fracasar al tratamiento con inmunosupresores, en la actualidad está en tratamiento con esteroides orales y tocilizumab. Los especialistas en oftalmología deben estar al corriente de la afectación oftalmológica de las enfermedades autoinflamatorias, y en especial de esta nueva entidad descrita, como es el síndrome VEXAS (AU)

VEXAS (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic) syndrome is a newly described autoinflammatory entity caused by a UBA-1 gene mutation. Among the most frequent symptoms it produces fever, cytopenias, polychondritis, pulmonary infiltrates and up to 40% ocular involvement such as periorbital edema, uveitis, episcleritis, scleritis and retinal vasculitis. Patients respond to high doses of corticosteroids, however, many end up being refractory to them and to the classic immunosuppressants. We described the case of a 77-year-old male patient with ocular involvement in the form of episcleritis and periorbital edema who was later diagnosed with VEXAS syndrome. The patient, after failing treatment with immunosuppressants, is currently receiving treatment with oral steroids and tocilizumab. Ophthalmologist must be aware of the ophthalmological affectation of autoinflammatory diseases and especially of this new entity described as the VEXAS syndrome (AU)

Episcleritis and periorbital edema secondary to VEXAS syndrome.

VEXAS (vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic) syndrome is a newly described autoinflammatory entity caused by a UBA-1 gene mutation. Among the most frequent symptoms it produces fever, cytopenias, polychondritis, pulmonary infiltrates and up to 40% ocular involvement such as periorbital edema, uveitis, episcleritis, scleritis and retinal vasculitis. Patients respond to high doses of corticosteroids, however, many end up being refractory to them and to the classic immunosuppressants. We described the case of a 77-year-old male patient with ocular involvement in the form of episcleritis and periorbital edema who was later diagnosed with VEXAS Syndrome. The patient, after failing treatment with immunosuppressants, is currently receiving treatment with oral steroids and tocilizumab. Ophthalmologist must be aware of the ophthalmological affectation of autoinflammatory diseases and especially of this new entity described as the VEXAS Syndrome.

Phylogenetic variations in a novel family of hyperstable apple snail egg proteins: insights into structural stability and functional trends.

The relationship between protein stability and function evolution has not been explored in proteins from natural sources. Here, we investigate the phylogenetic differences of Perivitellin-1 (PV1) a novel family of hyperstable egg carotenoproteins crucial to the reproductive success of Pomacea snails, as they have evolved clade-specific protective functions. We studied P. patula PV1 (PpaPV1) from Flagellata clade eggs, the most basal of Pomacea and compared it with PV1s orthologs from derived clades. PpaPV1 stands as the most stable, with longer unfolding half-life, resistance to detergent unfolding, and therefore higher kinetic stability than PV1s from derived clades. In fact, PpaPV1 is among the most hyperstable proteins described in nature. In addition, its spectral characteristics providing a pale egg coloration, mild lectin activity and glycan specificity are narrower than derived clades. Our results provide evidence indicating large structural and functional changes along the evolution of the genus. Notably, the lectin binding of PpaPV1 is less pronounced, and its glycan specificity is narrower compared to PV1s in the sister Bridgesii clade. Our findings underscore the phylogenetic disparities in terms of structural and kinetic stability, as well as defensive traits like a potent lectin activity affecting the gut morphology of potential predators within the Bridgesii clade or a conspicuous, likely warning coloration, within the Canaliculata clade. This work provides a comprehensive comparison of the structural attributes, stability profiles, and functional roles of apple snail egg PV1s from multiple species within a phylogenetic context. Furthermore, it proposes an evolutionary hypothesis suggesting a trade-off between structural stability and the functional aspects of apple snail's major egg defense protein.

Resultados del programa de cribado de enfermedad de Fabry en pacientes con córnea verticillata en el Hospital Universitario de Navarra / Results of the screening program for Fabry disease in patients with cornea verticillata at the University Hospital of Navarre

Introducción La cornea verticillata (CV) o queratopatía vortex se caracteriza por la presencia de depósitos en forma de espiral en el epitelio corneal. Las causas más frecuentes son los fármacos antipalúdicos y la amiodarona, y dentro de las causas sistémicas la enfermedad de Fabry (EF). Material y métodos Se realiza un estudio descriptivo prospectivo multidisciplinar en un centro de tercer nivel y de referencia en la Comunidad Foral de Navarra, tras la implementación de un programa de despistaje de la EF en los pacientes atendidos en consultas del servicio de oftalmología que eran diagnosticados de CV y posteriormente fueron remitidos a la consulta de enfermedades minoritarias del servicio de Medicina Interna del Hospital Universitario de Navarra para la realización del estudio de la EF. Resultados Dos mujeres y 4 varones, con una edad media de 76,8 años, se diagnosticaron de CV, de un total de 17.280 pacientes valorados en consultas externas por 3 especialistas de oftalmología durante el periodo de abril de 2018 a abril de 2020. Una paciente falleció antes de realizar el estudio de despistaje y en ningún paciente se diagnosticó la EF. Conclusiones A pesar de que el programa de despistaje de la EF en pacientes con CV en el Hospital Universitario de Navarra no confirmó ningún caso con EF, los especialistas en oftalmología deben tener en cuenta el posible diagnóstico de la EF en aquellos pacientes con CV en sus consultas rutinarias (AU)

Introduction Cornea verticillata (CV) or vortex keratopathy is characterized by the presence of spiral-shaped deposits in the corneal epithelium. The most frequent causes are antimalarial drugs and amiodarone and, among systemic causes, Fabry disease (FD). Material and methods A multidisciplinary, prospective, descriptive study was conducted in a tertiary reference center in Spain's Autonomous Community of Navarre after the implementation of a FD screening program for patients attended to in the Ophthalmology Department. The study analyzed those diagnosed with CV, who were subsequently referred to the rare disease clinic of the University Hospital of Navarre's Internal Medicine Department for an FD study. Results Two women and four men with a mean age of 76.8 years were diagnosed with CV out of a total of 17,280 patients evaluated in outpatient consultations by three ophthalmology specialists during the period from April 2018 to April 2020. One patient died before performing the screening study and no patients were diagnosed with FD. Conclusions Despite the fact that the University Hospital of Navarre's FD screening program for patients with CV did not confirm any cases of FD, ophthalmology specialists should consider the possible diagnosis of FD in patients with CV in their routine consultations (AU)

Results of the screening program for Fabry disease in patients with cornea verticillata at the University Hospital of Navarre.

INTRODUCTION: Cornea verticillata (CV) or vortex keratopathy is characterized by the presence of spiral-shaped deposits in the corneal epithelium. The most frequent causes are antimalarial drugs and amiodarone and, among systemic causes, Fabry disease (FD). MATERIAL AND METHODS: A multidisciplinary, prospective, descriptive study was conducted in a tertiary reference center in Spain's Autonomous Community of Navarre after the implementation of a FD screening program for patients attended to in the Ophthalmology Department. The study analyzed those diagnosed with CV, who were subsequently referred to the rare disease clinic of the University Hospital of Navarre's Internal Medicine Department for an FD study. RESULTS: Two women and four men with a mean age of 76.8 years were diagnosed with CV out of a total of 17,280 patients evaluated in outpatient consultations by three ophthalmology specialists during the period from April 2018 to April 2020. One patient died before performing the screening study and no patients were diagnosed with FD. CONCLUSIONS: Despite the fact that the University Hospital of Navarre's FD screening program for patients with CV did not confirm any cases of FD, ophthalmology specialists should consider the possible diagnosis of FD in patients with CV in their routine consultations.

[Impact of novel coronavirus infection in patients with uveitis associated with an autoimmune disease: result of the COVID-19-GEAS patient survey]. / Impacto de la infección por el nuevo coronavirus en los pacientes con uveítis asociada a una enfermedad autoinmune: resultado de la encuesta COVID-19-GEAS pacientes.

INTRODUCTION: The objetive of these study is to know the characteristics of COVID-19 in patients with uveitis associated with Systemic Autoimmune Disease (SAD) through telematic survey. MATERIAL AND METHODS: Internal Medicine Society and Group of Systemic Autoimmune disease conducted a telematic survey of patients with SAD to learn about the characteristics of COVID-19 in this population. RESULTS: A total of 2,789 patients answered the survey, of which 28 had a diagnosis of uveitis associated with SAE. The majority (82%) were female and caucasian (82%), with a mean age of 48 years. The most frequent SAEs were Behçet's disease followed by sarcoidosis and systemic lupus erythematosus. 46% of the patients were receiving corticosteroid treatment at a mean prednisone dose of 11 mg/day. Regarding infection, 14 (50%) patients reported symptoms compatible with SARS-CoV-2 infection. RT-PCR was performed on the nasopharyngeal smear in two patients and in one of them (4%) it was positive. CONCLUSIONS: Both asymptomatic and symptomatic COVID-19 patients with ASD-associated UNI had received similar immunosuppressive treatment.

Impact of novel coronavirus infection in patients with uveitis associated with an autoimmune disease: Result of the COVID-19-GEAS patient survey.

INTRODUCTION: The objective of these study is to know the characteristics of COVID-19 in patients with uveitis associated with Systemic Autoimmune Disease (SAD) through telematic survey. MATERIAL AND METHODS: Internal Medicine Society and Group of Systemic Autoimmune disease conducted a telematic survey of patients with SAD to learn about the characteristics of COVID-19 in this population. RESULTS: A total of 2,789 patients answered the survey, of which 28 had a diagnosis of uveitis associated with SAE. The majority (82%) were female and caucasian (82%), with a mean age of 48 years. The most frequent SAEs were Behçet's disease followed by sarcoidosis and systemic lupus erythematosus. 46% of the patients were receiving corticosteroid treatment at a mean prednisone dose of 11 mg/day. Regarding infection, 14 (50%) patients reported symptoms compatible with SARS-CoV-2 infection. RT-PCR was performed on the nasopharyngeal smear in two patients and in one of them (4%) it was positive. CONCLUSIONS: Both asymptomatic and symptomatic COVID-19 patients with ASD-associated UNI had received similar immunosuppressive treatment.

Impacto de la infección por el nuevo coronavirus en los pacientes con uveítis asociada a una enfermedad autoinmune: resultado de la encuesta COVID-19-GEAS pacientes / Impact of novel coronavirus infection in patients with uveitis associated with an autoimmune disease: result of the COVID-19-GEAS patient survey

Introducción El objetivo de este estudio es conocer las características de la COVID-19 en pacientes con uveítis asociada a enfermedades autoinmunes sistémicas (EAS) mediante una encuesta telemática. Material y métodos La Sociedad Española de Medicina Interna por medio del Grupo de Trabajo de Enfermedades Autoinmunes realizó una encuesta telemática a pacientes con EAS para conocer las características de la COVID-19 en esta población. Resultados Un total de 2.789 pacientes contestaron la encuesta, de los que 28 tenían un diagnóstico de uveítis asociada a una EAS. La mayoría (82%) eran mujeres y caucásicas (82%), con una media de 48 años. Las EAS más frecuentes fueron la enfermedad de Behçet seguida de la sarcoidosis y del lupus eritematoso sistémico. El 46% de los pacientes estaban recibiendo tratamiento con corticoides a una dosis media de prednisona de 11mg/día. Respecto a la infección, 14 (50%) pacientes referían síntomas compatibles con infección por SARS-CoV-2. Se realizó RT-PCR en el frotis nasofaríngeo en dos pacientes y en uno de ellos (4%) fue positivo. Conclusiones Los pacientes con UNI asociada a EAS tanto los asintomáticos como los sintomáticos de COVID-19 habían recibido de forma similar tratamiento inmunosupresor (AU)

Introduction The objetive of these study is to know the characteristics of COVID-19 in patients with uveitis associated with Systemic Autoimmune Disease (SAD) through telematic survey. Material and methods Internal Medicine Society and Group of Systemic Autoimmune disease conducted a telematic survey of patients with SAD to learn about the characteristics of COVID-19 in this population. Result a total of 2,789 patients answered the survey, of which 28 had a diagnosis of uveitis associated with SAE. The majority (82%) were female and caucasian (82%), with a mean age of 48 years. The most frequent SAEs were Behçet's disease followed by sarcoidosis and systemic lupus erythematosus. 46% of the patients were receiving corticosteroid treatment at a mean prednisone dose of 11mg/day. Regarding infection, 14 (50%) patients reported symptoms compatible with SARS-CoV-2 infection. RT-PCR was performed on the nasopharyngeal smear in two patients and in one of them (4%) it was positive. Conclusions Both asymptomatic and symptomatic COVID-19 patients with ASD-associated UNI had received similar immunosuppressive treatment (AU)

Exaptation of two ancient immune proteins into a new dimeric pore-forming toxin in snails.

The Membrane Attack Complex-Perforin (MACPF) family is ubiquitously found in all kingdoms. They have diverse cellular roles, however MACPFs with pore-forming toxic function in venoms and poisons are very rare in animals. Here we present the structure of PmPV2, a MACPF toxin from the poisonous apple snail eggs, that can affect the digestive and nervous systems of potential predators. We report the three-dimensional structure of PmPV2, at 17.2 Å resolution determined by negative-stain electron microscopy and its solution structure by small angle X-ray scattering (SAXS). We found that PV2s differ from nearly all MACPFs in two respects: it is a dimer in solution and protomers combine two immune proteins into an AB toxin. The MACPF chain is linked by a single disulfide bond to a tachylectin chain, and two heterodimers are arranged head-to-tail by non-covalent forces in the native protein. MACPF domain is fused with a putative new Ct-accessory domain exclusive to invertebrates. The tachylectin is a six-bladed ß-propeller, similar to animal tectonins. We experimentally validated the predicted functions of both subunits and demonstrated for the first time that PV2s are true pore-forming toxins. The tachylectin "B" delivery subunit would bind to target membranes, and then the MACPF "A" toxic subunit would disrupt lipid bilayers forming large pores altering the plasma membrane conductance. These results indicate that PV2s toxicity evolved by linking two immune proteins where their combined preexisting functions gave rise to a new toxic entity with a novel role in defense against predation. This structure is an unparalleled example of protein exaptation.

Complicaciones y agudeza visual en los pacientes con uveítis en una unidad multidisciplinar de referencia del norte de España / Complications and visual acuity of patients with uveitis: Epidemiological study in a reference unit in northern Spain

Objetivo: Describir las complicaciones y la agudeza visual en una cohorte de 500 pacientes diagnosticados en una unidad multidisciplinar de referencia en el norte de España. Material y métodos: Se realizó un estudio retrospectivo-prospectivo de las complicaciones y de la agudeza visual de 500 pacientes adultos con uveítis evaluados en la Unidad Multidisciplinar del Complejo Hospitalario de Navarra desde enero de 2010 hasta marzo de 2015. Se analizó el tipo de complicación, la agudeza visual y el grado de pérdida de agudeza visual al año de seguimiento y se realizó un estudio comparativo de las complicaciones con otros 2 estudios previos publicados en Madrid y en Holanda. Resultados. La pérdida de agudeza visual moderada-severa fue del 13,5% en el ojo derecho y del 13% en el ojo izquierdo. El grupo de edad de 65 años o superior se asoció a mayor pérdida de agudeza visual. El 35% de los pacientes presentaron alguna complicación al año de seguimiento, y las complicaciones más frecuentes fueron la catarata (10%), las sinequias (8%) y el edema macular (5%). En comparación con las otras series, en Navarra la catarata fue la complicación más frecuente. Conclusiones: La pérdida de agudeza visual se asoció con la mayor edad de los pacientes y la catarata fue la complicación más frecuente en nuestro estudio

Objective: To describe uveitis complications and visual acuity in a cohort of 500 patients in a multidisciplinary unit in northern Spain. Material and methods: Retrospective-prospective study of complications and visual acuity of 500 adult patients evaluated in the Multidisciplinary Unit of the Navarra Hospital Complex from the period January 2010 until March 2015. An analysis was made of the complications, visual acuity and visual loss, with a follow-up of one-year. A comparative study was also made of the complications with 2 previous series published in Madrid and Holland. Results: Moderate-severe visual loss was 13.5% in the right eye, and 13% in the left eye. Visual loss was associated with an age of 65 years or above. Complications were observed in 35% of patients, and cataract was the most frequent complication (10%), followed by synechiae (8%), and macular oedema (5%). Compared with the 2 other series, the present cohort showed a higher proportion of cataracts. Conclusions: Visual loss was associated with older age and cataract was the most common complication in our study

Complications and visual acuity of patients with uveitis: Epidemiological study in a reference unit in northern Spain. / Complicaciones y agudeza visual en los pacientes con uveítis en una unidad multidisciplinar de referencia del norte de España.

OBJECTIVE: To describe uveitis complications and visual acuity in a cohort of 500 patients in a multidisciplinary unit in northern Spain. MATERIAL AND METHODS: Retrospective-prospective study of complications and visual acuity of 500 adult patients evaluated in the Multidisciplinary Unit of the Navarra Hospital Complex from the period January 2010 until March 2015. An analysis was made of the complications, visual acuity and visual loss, with a follow-up of one-year. A comparative study was also made of the complications with 2 previous series published in Madrid and Holland. RESULTS: Moderate-severe visual loss was 13.5% in the right eye, and 13% in the left eye. Visual loss was associated with an age of 65 years or above. Complications were observed in 35% of patients, and cataract was the most frequent complication (10%), followed by synechiae (8%), and macular oedema (5%). Compared with the 2 other series, the present cohort showed a higher proportion of cataracts. CONCLUSIONS: Visual loss was associated with older age and cataract was the most common complication in our study.

Biosynthesis in the Albumen Gland-Capsule Gland Complex Limits Reproductive Effort in the Invasive Apple Snail Pomacea canaliculata.

High fecundity often contributes to successful invasives. In molluscs, this may be facilitated by the albumen gland-capsule gland complex, which in gastropods secretes the egg perivitelline fluid that nourishes and protects embryos. The biochemistry of the albumen gland-capsule gland complex and its relationship with fecundity remain largely unknown. We addressed these issues in Pomacea canaliculata (Lamarck, 1822), a highly invasive gastropod whose fecundity and reproductive effort exceed those of ecologically similar gastropods. We evaluated the dynamics of its major secretion compounds (calcium, polysaccharides, and total proteins) as well as the gene expression and stored levels of perivitellins during key moments of the reproductive cycle, that is, before and after first copulation and at low, medium, and high reproductive output. Copulation and first oviposition do not trigger the onset of albumen gland-capsule gland complex biosynthesis. On the contrary, soon after an intermediate reproductive effort, genes encoding perivitellins overexpressed. A high reproductive effort caused a decrease in all albumen gland-capsule gland complex secretion components. Right after a high reproductive output, the albumen gland-capsule gland complex restored the main secretion components, and calcium recovered baseline reserves; but proteins and polysaccharides did not. These metabolic changes in the albumen gland-capsule gland complex after multiple ovipositions were reflected in a reduction in egg mass but did not compromise egg quality. At the end of the cycle, egg dry weight almost doubled the initial albumen gland-capsule gland complex weight. Results indicate that albumen gland-capsule gland complex biosynthesis limits a constantly high reproductive output. Therefore, lowering fecundity by targeting biosynthesis could effectively reduce the rate of this species' spread.

Caracterización de los pacientes con uveítis remitidos a una unidad multidisciplinar de referencia en el norte de España / Profile of patients with uveitis referred to a multidisciplinary unit in northern Spain

OBJETIVO: Describir las características de los pacientes con uveítis valorados en una unidad multidisciplinar de referencia del norte de España. MATERIAL Y MÉTODOS: Análisis retrospectivo de los pacientes con uveítis valorados en la Unidad Multidisciplinar del Complejo Hospitalario de Navarra desde enero del 2010 hasta marzo del 2015. Se analizaron las características demográficas, procedencia, tipos de uveítis, lateralidad, etiología y se compararon estas características con las de 2 series de pacientes de Castilla y León y de Barcelona. RESULTADOS: Se analizó a un total de 500 pacientes, 50% mujeres y con una edad media de 47,9±16,4 años. El 65,4% de las uveítis fueron anteriores; el 17,6%, posteriores; el 15,2%, panuveítis y el 1,8%, de localización intermedia. El 31,2% fueron no clasificables, seguidas de la enfermedad sistémica no infecciosa con el 29,2%. La espondilitis anquilopoyética (10,8%), la infección por virus herpes (9,2%) y la toxoplasmosis (7,8%) fueron las 3causas más frecuentes. En comparación con las otras 2series, en la de este estudio se encontró una mayor proporción de uveítis anteriores unilaterales. Además, en Navarra los pacientes presentaron mayor prevalencia de uveítis anterior unilateral y de uveítis idiopáticas en relación con la serie de Barcelona. CONCLUSIONES: Los datos de nuestro estudio demuestran características similares con las uveítis de otras regiones de nuestro país. Las uveítis anteriores unilaterales y las idiopáticas fueron las más frecuentes en nuestra serie

OBJECTIVE: To describe the main characteristics of a cohort of patients with uveitis referred to a multidisciplinary unit in northern Spain. MATERIAL AND METHODS: Retrospective analysis of clinical records of patients evaluated in the Multidisciplinary Unit of the Navarra Hospital Complex from the period January 2010 until March 2015. An analysis was performed on the demographic characteristics, origin, types of uveitis, laterality, and aetiology. The present series was also compared with 2 previous series from Castilla y León and Barcelona. RESULTS: A total of 500 patients were identified, with a mean age of 47.9±16.4 years, with 50% women. The most frequent type of uveitis was anterior uveitis (65.4%), followed by posterior uveitis (17.6%), panuveitis (15.2%), and intermediate uveitis (1.8%). The origin was unclassifiable in 31.2%, followed by non-infectious systemic disease in 29.2%. Ankylosing spondylitis was the most frequent cause in 10.8% of patients, followed by herpes infection in 9.2%, and toxoplasmosis in 7.8%, respectively. Compared with the 2other cohorts, the present cohort showed a higher proportion of unilateral anterior uveitis. Furthermore, the patients from the Navarra series had a higher prevalence of unilateral and idiopathic uveitis compared to the series from Barcelona. CONCLUSIONS: The main characteristics of the present cohort of patients with uveitis are similar to those of patients from other regions of our country. Unilateral anterior uveitis and idiopathic uveitis were the most frequent in our series

Profile of patients with uveitis referred to a multidisciplinary unit in northern Spain. / Caracterización de los pacientes con uveítis remitidos a una unidad multidisciplinar de referencia en el norte de España.

OBJECTIVE: To describe the main characteristics of a cohort of patients with uveitis referred to a multidisciplinary unit in northern Spain. MATERIAL AND METHODS: Retrospective analysis of clinical records of patients evaluated in the Multidisciplinary Unit of the Navarra Hospital Complex from the period January 2010 until March 2015. An analysis was performed on the demographic characteristics, origin, types of uveitis, laterality, and aetiology. The present series was also compared with 2 previous series from Castilla y León and Barcelona. RESULTS: A total of 500 patients were identified, with a mean age of 47.9±16.4 years, with 50% women. The most frequent type of uveitis was anterior uveitis (65.4%), followed by posterior uveitis (17.6%), panuveitis (15.2%), and intermediate uveitis (1.8%). The origin was unclassifiable in 31.2%, followed by non-infectious systemic disease in 29.2%. Ankylosing spondylitis was the most frequent cause in 10.8% of patients, followed by herpes infection in 9.2%, and toxoplasmosis in 7.8%, respectively. Compared with the 2other cohorts, the present cohort showed a higher proportion of unilateral anterior uveitis. Furthermore, the patients from the Navarra series had a higher prevalence of unilateral and idiopathic uveitis compared to the series from Barcelona. CONCLUSIONS: The main characteristics of the present cohort of patients with uveitis are similar to those of patients from other regions of our country. Unilateral anterior uveitis and idiopathic uveitis were the most frequent in our series.

The major egg reserve protein from the invasive apple snail Pomacea maculata is a complex carotenoprotein related to those of Pomacea canaliculata and Pomacea scalaris.

Snails from the genus Pomacea lay conspicuous masses of brightly colored eggs above the water. Coloration is given by carotenoproteins that also which play important roles in protection against sun radiation, stabilizing and transporting antioxidant molecules and helping to protect embryos from desiccation and predators. They seem a key acquisition, but have been little studied. Here we report the characteristics of the major carotenoprotein from Pomacea maculata and the first comparison among these egg proteins. This particle, hereafter PmPV1, represents ~52% of perivitellin fluid protein. It is a glyco-lipo-carotenoprotein responsible for the bright reddish egg coloration. With VHDL characteristics, PmPV1 apparent molecular mass is 294kDa, composed of five non-covalently bound subunits of pI 4.7-9.8 and masses between 26 and 36kDa whose N-terminal sequences were obtained. It is a glyco-lipo-carotenoprotein scarcely lipidated (<1%) but highly glycosilated (13% by wt). Lipids include phospholipids, free fatty acids and carotenoids; mannose and galactose predominate over other monosaccharides. Main carotenoids are esterified and non-esterified astaxanthin (71 and 25%, respectively). Carotenoid removal does not seem to affect the structural characteristics of the oligomer, while deglycosilation reduces subunit number from five to a single one. The carotenoid-protein association protected the former against oxidation. PmPV1 cross reacts with polyclonal antibodies against the PcOvo, the major carotenoprotein from Pomacea canaliculata. The characterization of PmPV1 allows the first comparisons among snail carotenoproteins and further highlights the importance of these perivitellins in the reproductive strategy of Pomacea.

Study of biochemical biomarkers in freshwater prawn Macrobrachium borellii (Crustacea: Palaemonidae) exposed to organophosphate fenitrothion.

Several agrochemicals like organophosphates are extensively used to control pests in agricultural practices but they also adversely affect non-target fauna. The effect of organophosphorous fenitrothion on the prawn Macrobrachium borellii was evaluated. The 96-h LC50 was determined. Activity of superoxide dismutase, catalase, glutathione-S-transferase and lipid oxidation levels, were evaluated in the hepatopancreas from adults exposed to sublethal fenitrothion concentrations for 1, 2, 4 and 7 days. In addition, superoxide dismutase mRNA expression, acetylcholinesterase inhibition and haemocyte DNA damage were determined. The 96-h LC50 was 4.24µg/l of fenitrothion. Prawn exposed to sublethal FS concentrations showed an increase of both catalase and superoxide dismutase activities, mainly after 2 and 4 days exposure and an increase of glutathione-S-transferase activity from day 2 to day 7 while lipid oxidation levels increased mainly on day 1. Superoxide dismutase transcripts were significantly higher in fenitrothion -treated prawns, indicating an induction mechanism. Hemolymph analysis showed that while acetylcholinesterase activity decreased after 2 days, haemocytes displayed most DNA damage after 7-day exposure to fenitrothion. These results indicate that prawn enzymes are highly sensitive to fenitrothion exposure, and these biological responses in M. borellii could be valuable biomarkers to monitor organophosphorous contamination in estuarine environments.

Effect of crude oil petroleum hydrocarbons on protein expression of the prawn Macrobrachium borellii.

Hydrocarbon pollution is a major environmental threat to ecosystems in marine and freshwater environments, but its toxicological effect on aquatic organisms remains little studied. A proteomic approach was used to analyze the effect of a freshwater oil spill on the prawn Macrobrachium borellii. To this aim, proteins were extracted from midgut gland (hepatopancreas) of male and female prawns exposed 7 days to a sublethal concentration (0.6 ppm) of water-soluble fraction of crude oil (WSF). Exposure to WSF induced responses at the protein expression level. Two-dimensional gel electrophoresis (2-DE) revealed 10 protein spots that were differentially expressed by WSF exposure. Seven proteins were identified using MS/MS and de novo sequencing. Nm23 oncoprotein, arginine methyltransferase, fatty aldehyde dehydrogenase and glutathione S-transferase were down-regulated, whereas two glyceraldehyde-3-phosphate dehydrogenase isoforms and a lipocalin-like crustacyanin (CTC) were up-regulated after WSF exposure. CTC mRNA levels were further analyzed by quantitative real-time PCR showing an increased expression after WSF exposure. The proteins identified are involved in carbohydrate and amino acid metabolism, detoxification, transport of hydrophobic molecules and cellular homeostasis among others. These results provide evidence for better understanding the toxic mechanisms of hydrocarbons. Moreover, some of these differentially expressed proteins would be employed as potential novel biomarkers.

Triacylglycerol catabolism in the prawn Macrobrachium borellii (Crustacea: Palaemoniade).

While invertebrates store neutral lipids as their major energy source, little is known about triacylglycerol (TAG) class composition and their differential catabolism in aquatic arthropods. This study focuses on the composition of the main energy source and its catabolism by lipase from the midgut gland (hepatopancreas) of the crustacean Macrobrachium borellii. Silver-ion thin-layer chromatography of prawn large TAG deposit (80% of total lipids) and its subsequent fatty acid analysis by gas chromatography allowed the identification of 4 major fractions. These are composed of fatty acids of decreasing unsaturation and carbon chain length, the predominant being 18:1n-9. Fraction I, the most unsaturated one, contained mainly 20:5n-3; fraction II 18:2n-6; fraction III 18:1n-9 while the most saturated fraction contained mostly 16:0. Hepatopancreas main lipase (Mr 72 kDa) cross-reacted with polyclonal antibodies against insect lipase, was not dependent on the presence of Ca(2+) and had an optimum activity at 40°C and pH 8.0. Kinetic analysis showed a Michaelis-Menten behavior. A substrate competition assay evidenced lipase specificity following the order: 18:1n-9-TAG>PUFA-enriched-TAG>16:0-TAG different from that in vertebrates. These data indicate there is a reasonable correspondence between the fatty acid composition of TAG and the substrate specificity of lipase, which may be an important factor in determining which fatty acids are mobilized during lipolysis for oxidation in crustaceans.

Antioxidant response and oxidative stress levels in Macrobrachium borellii (Crustacea: Palaemonidae) exposed to the water-soluble fraction of petroleum.

The aim of the present work was to evaluate the effect of the water soluble fraction of hydrocarbons (WSF) on the antioxidant status of the freshwater prawn Macrobrachium borellii. First, seasonal variations were studied in a non-polluted area. Hepatopancreas and gills showed season-related fluctuations in catalase (CAT), glutathione-S-transferase (GST) activities and in lipid peroxidation levels (LPO), but not in superoxide dismutase (SOD). Then, adults were exposed semi-statically to sublethal doses for 7days. CAT, SOD, GST, and glutathione peroxidase (GPx) activities and LPO, reduced glutathione (GSH) and protein oxidation (PO) levels were determined. Exposed individuals showed significant increases in CAT, SOD, and GST activities in hepatopancreas and CAT activity in gills. GPx activity did not vary in either tissues. While LPO levels increased, GSH levels decreased significantly in hepatopancreas of exposed animals, but PO levels showed no variation. Induction of SOD was also assessed by Real-time PCR mRNA expression in hepatopancreas. The non-enzymatic antioxidant activity was also tested; ABTS 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid) was higher in hemolymph of treated-prawns compared to controls, but ferric reducing activity of plasma assay (FRAP) values did not change. Taken together, the present results indicated that the antioxidant defenses of M. borellii, mainly in hepatopancreas, were significantly affected by aquatic hydrocarbon contamination, regardless of the season.

Carbohydrates and glycoforms of the major egg perivitellins from Pomacea apple snails (Architaenioglossa: Ampullariidae).

To better understand how glycans contribute to the multiple roles of perivitellins in embryo development, the carbohydrate moieties and glycoforms of the carotenoglycoproteins ovorubin and scalarin from the eggs of Pomaceacanaliculata (Lamarck, 1822) and Pomaceascalaris (d'Orbigny, 1835) were studied. All subunits of both proteins are glycosylated and appear to be glycoforms with isoelectric points ranging from approximately 5.3 to approximately 9.1. Complete deglycosylation reduced subunit heterogeneity to spots of similar molecular weight (approximately 27 and approximately 25 kDa in scalarin and ovorubin, respectively) but with varying IP. Serine phosphorylation, present in both perivitellins, explains part of the isoforms. Glycosylation patterns of scalarin were determined using biotinylated lectins, PNGaseF treatment and selective chemical deglycosylation, which revealed the presence of hybrid and oligomannose N-linked glycans in all subunits. Scalarin has terminal sialic acid residues possibly resistant to neuraminidase and O-linked residues derived from the T- and Tn antigens. Ovorubin displayed predominantly the same glycans, though in different amounts. Capillary gas chromatography (GC) showed galactose and mannose as the major monosaccharides followed by GlcNAc and fucose. An interesting feature was the important amount of sialylated and fucosylated structures found in both perivitellins determined by GC, spectroscopy and lectins. This is the first report of these structures in gastropods other than heterobranchs.