Immunization of sea bream (Sparus aurata) juveniles against Photobacterium damselae subsp. piscicida by short bath: Effect on some pro-inflammatory molecules and the Mx gene expression.

Cytokines are a family of proteins derived from macrophages, lymphocytes, granulocytes, mast cells and epithelial cells and can be divided into interferons (IFNs), Interleukins (ILs) and Tumor Necrosis factors (TNFs) among others. The presence of cytokines in a wide number of fish species has been proved and several molecules types have been already cloned and sequenced. In this work some proinflamatory molecules and Mx gene were detected in the liver of vaccinated sea bream juveniles with an average body weight of 5 g. The method of immunization was by short bath and three different bacterins against the marine pathogen Photobacterium damselae subsp. piscicida were designed and used to immunize fish. Five genes encoding for five different molecules were analyzed by real time PCR: IL-1ß, IL Ir-2, Cox-2, Mx and TNFα. Gene expression was quantified along four days after fish immunization and results were compared among groups. Results show that the heat-inactivated vaccine stimulates the up-regulation of IL-1ß, IL Ir-2, Cox-2 and TNFα genes whereas the UV-light inactivated vaccine was the unique vaccine which stimulates the expression of Mx gene. The present is a novel study that shows by the first time the effect of the inactivation process of vaccines on the expression levels of genes involved in the defense against Photobacterium damselae subsp piscicida.

Síndrome de insulinorresistencia severa tipo A debido a mutación del gen del receptor de insulina / Severe type A insulin resistance syndrome due to a mutation in the insulin receptor gene

Los síndromes monogénicos de insulinorresistencia sin lipodistrofia constituyen un rupo de entidades infrecuentes que incluyen los síndromes de Donohue o leprechaunismo, Rabson-Mendenhall y resistencia a la insulina tipo A. Se caracterizan por un amplio espectro fenotípico que asocia insulinorresistencia extrema y alteraciones hidrocarbonadas de grado variable. Presentamos un caso de resistencia a la insulina tipo A, caracterizado por resistencia insulínica grave, acantosis nigricans e hiperandrogenismo, debido a una mutación en heterocigosis en el xón 19 del gen del receptor de insulina que codifica para el dominio tirosinquinasa. Se destaca la elevada morbilidad de dicha entidad, a pesar de incluirse dentro del espectro menos grave de los síndromes genéticos de resistencia insulínica, así como la ausencia de una terapia satisfactoria. El estudio molecular revela el diagnóstico e informa del pronóstico y la supervivencia, factores ligados a la función residual del receptor, además de contribuir al desarrollo de nuevas dianas terapéuticas

Insulin resistance syndromes without lipodystrophy are an infrequent and heterogeneous group of disorders with variable clinical phenotypes, associated with hyperglycemia and hyperinsulinemia. The three conditions related to mutations in the insulin receptor gene are leprechaunism or Donohue syndrome, Rabson-Mendenhall syndrome, and Type A syndrome. A case is presented on a patient diagnosed with type A insulin resistance, defined by the triad of extreme insulin resistance, acanthosis nigricans, and hyperandrogenism, carrying a heterozygous mutation in exon 19 of the insulin receptor gene coding for its tyrosine kinase domain that is crucial for the catalytic activity of the receptor. The molecular basis of the syndrome is reviewed, focusing on the structure-function relationships of the insulin receptor, knowing that the criteria for survival are linked to residual insulin receptor function. It is also pointed out that, although type A insulin resistance appears to represent a somewhat less severe condition, these patients have a high morbidity and their treatment is still unsatisfactory

[Severe type A insulin resistance syndrome due to a mutation in the insulin receptor gene]. / Síndrome de insulinorresistencia severa tipo A debido a mutación del gen del receptor de insulina.

Insulin resistance syndromes without lipodystrophy are an infrequent and heterogeneous group of disorders with variable clinical phenotypes, associated with hyperglycemia and hyperinsulinemia. The three conditions related to mutations in the insulin receptor gene are leprechaunism or Donohue syndrome, Rabson-Mendenhall syndrome, and Type A syndrome. A case is presented on a patient diagnosed with type A insulin resistance, defined by the triad of extreme insulin resistance, acanthosis nigricans, and hyperandrogenism, carrying a heterozygous mutation in exon 19 of the insulin receptor gene coding for its tyrosine kinase domain that is crucial for the catalytic activity of the receptor. The molecular basis of the syndrome is reviewed, focusing on the structure-function relationships of the insulin receptor, knowing that the criteria for survival are linked to residual insulin receptor function. It is also pointed out that, although type A insulin resistance appears to represent a somewhat less severe condition, these patients have a high morbidity and their treatment is still unsatisfactory.

Tyrosine kinase inhibitors: muco-cutaneous side effects at the microscope.

In the past recent years, treatments that target receptors with kinase activity, involved in the transmission of neoplastic proliferation signals, had revolutionized cancer therapy. Imatinib mesylate has been the first of this novel family of drugs approved for the treatment of hematologic malignancies. Afterwards, other second-generation kinase inhibitors, such as dasatinib and nilotinib, have been introduced to circumvent resistance to imatinib. These target therapies have a better tolerability profile than standard chemotherapy, but their range of activity is not simply directed at tumor cells, and a wide spectrum of systemic side effects is now recognized. In particular, muco-cutaneous side effects represent the most frequent non-hematological adverse events. Due to the need of a prompt recognition of these toxicities, diagnosis is usually made on clinical grounds, and an accurate histological characterization is generally lacking. The aim of this paper was to focus on the histopathological findings of cutaneous reactions related to tyrosine kinase inhibitors use. We propose a differentiation between specific and non-specific cutaneous side effects, through an analysis of the possible etiopathogenetic mechanisms of actions of the drug, clinical aspects and major histological features. A review of the literature has been integrated by our personal experience, highlighting the importance of clinico-histological correlation, necessary to make a proper diagnosis.

Immunogenetic factors in mycosis fungoides: can the HLA system influence the susceptibility and prognosis of the disease? Long-term follow-up study of 46 patients.

BACKGROUND: Mycosis fungoides (MF) is the most common and one of the least aggressive forms of cutaneous T-cell lymphoma. Several studies have demonstrated the influence of human leucocyte antigen (HLA) genes on the susceptibility of MF, highlighting the importance of certain alleles but, until today, no studies have evaluated the relationship between HLA alleles and the prognosis of patients with MF. OBJECTIVE: The aim of this retrospective cohort study was to evaluate the polymorphism of HLA class I and class II alleles in a group of 46 MF Caucasian patients, looking for their influence in susceptibility and prognosis of the disease. METHODS: Study population included a case-cohort sample of 46 Caucasian patients with MF that, between 1993 and 1997, underwent HLA class I and II genomic typing. All patients were diagnosed and followed up from 1977 to 2012 (mean follow-up of 11 years) and they were divided into three groups according to the evolution of the disease. RESULTS: Molecular typing at low-resolution level revealed that HLA-A*24, A*68, A*69, B*35 and DQB1*05:02 alleles were involved in susceptibility to MF. Correspondence analysis underlined that long-lasting remission was characterized by HLA-A*24 and HLA-A*25 alleles, frequent relapse by HLA-DRB1*01, DQA1*01:01, DQB1*05:01 alleles and death by HLA-A*68, HLA-B*08, HLA-B*35, HLA-C*03 alleles. CONCLUSION: This study suggests that the prognosis of MF patients is not only correlated with clinical/pathological/serological/immunological variables but it also relies on specific HLA alleles.

Cytokine expression in head-kidney leucocytes of European sea bass (Dicentrarchus labrax L.) after incubation with the probiotic Vagococcus fluvialis L-21.

The European sea bass (Dicentrarchus labrax L.) is one of the most extensively farmed marine fish in the Mediterranean sea. Under the high-density condition, common in aquaculture, the infectious diseases can cause significant economic losses. Probiotics are presented as an alternative to antibiotics for the control of aquaculture diseases. This study used real-time PCR to investigate in vitro the dynamic of expression of immune-related genes in sea bass after incubation with live and inactivated (heat and Uv-light) probiotic Vagoccus fluvialis L-21 at different times (T1, T12, T24, T48). The immune associated genes, interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin 10 (IL-10), TumourTumour necrosis factor- (TNF-), ciclo-oxigenase-2 (COX-2), caspase-3 (Casp-3) and Mx were studied in head-kidney (HK) leucocytes of sea bass after incubation with the probiotic strain. Transcript of proinflammatory cytokines (IL-1, TNF-, COX-2) was highly up-regulated after 1 h of incubation with the probiotic strain V. fluvialis L-21. We found statistically significant difference in pick of expression of TNF-, after 1 h of incubation with Uv-light inactivated probiotic strain. The COX-2 expression was highly up-regulated at all times studied, with the exception of 12 and 24 h post incubation for the Uv-light inactivated bacteria. Transcript of IL-10 and Casp-3 showed the higher statistically significant differences of expression after 48 h post incubation with live bacteria. In the contrast, sea bass HK leucocytes expressed Mx at 12 and 48 h without statistically differences among treatments. Our results suggest that V. fluvialis L-21 is able to stimulate in vitro some immune-related genes associated with the early inflammatory response. Future studies in vivo are necessary to clarify this process in sea bass.

Imatinib, dasatinib and nilotinib: a review of adverse cutaneous reactions with emphasis on our clinical experience.

In the last years, several tyrosine kinase inhibitors (TKIs) have been developed and approved for human cancer treatment. Imatinib mesylate was the first of this novel family of drugs that target cancer-specific molecules and signalling pathways. The appearance of imatinib resistances led to the introduction of second-generation TKIs with higher potency and selectivity, such as dasatinib and nilotinib. However, the range of activity of these agents is not simply directed at tumour cells. Patients and their clinicians are indeed frequently confronted with the cutaneous side-effects associated with the employ of these drugs, which represent the most common non-hematological adverse reactions. For this reason, a systematic dermatological survey of patients receiving these therapies is highly important, and an early and appropriate dermatological treatment is required. In this review, we analyse the clinical and pathological characteristics of the most commonly reported adverse skin events associated with first- and second-generation tyrosine kinase inhibitors, with a particular emphasis on our clinical experience.

Effect of lipopolysaccharides from Vibrio alginolyticus on the Mx gene expression and virus recovery from gilthead sea bream (Sparus aurata L.) experimentally infected with Nodavirus.

Infections with nodavirus affect a wild and farmed fish species throughout the world, mostly from the marine environment. The aim of this work was to determine the immune status of gilthead sea bream that comes as a result of a Nodavirus infection, induced by activation of the interferon response pathway by lipopolysaccharides from Vibrio alginolyticus and the expression of interferoninduced Mx protein in liver samples. The enhancement of Mx protein gene expression was detected in liver samples of experimentally nodavirus infected fish and, furthermore, the immunostimulant LPS of V. alginolyticus decreased almost three times the virus titration with respect to no-immunized or infected with nodavirus group of fish.

The in vitro effect of probiotic Vagococcus fluvialis on the innate immune parameters of Sparus aurata and Dicentrarchus labrax.

In this study we evaluated the effect of the probiotic Vagococcus fluvialis on the cellular immune unspecific system of two different fish species of great interest in aquaculture such as gilthead sea bream (Sparus aurata) and European sea bass (Dicentrarchus labrax). Leucocytes from head kidney of the two fish species were extracted and concentration adjusted to 10(7) cells ml(-1). Phagocytic and respiratory burst activity and the peroxidase content of leucocytes were observed 30 min after incubation with the probiotic Vagococcus fluvialis alive or inactivated with heat shock or UV-light at different concentrations of 10(7), 10(8), 10(9) cfu ml(-1) (final concentration 10(6), 10(7), 10(8) cfu ml(-1)). V. fluvialis produced dose-dependent increments in respiratory burst in sea bream leucocytes. The respiratory burst activity of sea bream head kidney leucocytes incubated with 10(6) cfu ml(-1) of live and inactivated bacteria was not stimulated. The highest values of peroxidase content were observed in sea bass cells with stimulation indexes higher than 1 in HK leucocytes incubated with 10(8) cfu ml(-1) of live and inactivated bacteria. Statistical analysis revealed that differences being only significant in sea bass leucocytes where 10(8) cfu ml(-1) bacteria denote statistically significant differences (P < 0.05) respect to other concentrations. Highest values of phagocytic activity were obtained in sea bass macrophages incubated with UV-light inactivated bacteria (27.33% ± 1.45), where significantly differences with sea bass HK leucocytes were detected. Our results suggest that the in vitro assays are a useful tool to optimize the effective dose of probiotic bacteria. Although in vivo studies are necessary to confirm the immunomodulatory effect of this strain.

Indolent systemic mastocytosis treated with narrow-band UVB phototherapy: study of five cases.

BACKGROUND: Mastocytoses represent a heterogeneous group of stem cell disorders marked by an abnormal hyperplasia and accumulation of mast cells in one or more tissues, including bone marrow, gastrointestinal tract, liver, spleen, lymph nodes and skin. Indolent systemic mastocytosis (ISM) is characterized by red-brownish and pruriginous maculopapular lesions, a bone marrow infiltration without functional impairment and an indolent clinical course with a good prognosis. In particular, the most common cutaneous symptoms are urticarial rash and mild-to-high pruritus. OBJECTIVES: This study analyses the clinical outcome of patients affected by ISM with prevalent pruriginous cutaneous symptoms and a scarce response to anti-histamines treated using narrowband ultraviolet B (NB-UVB) phototherapy. METHODS: Narrowband ultraviolet B phototherapy was administered in a UV-irradiation cabin equipped with fluorescent UVB lamps with a peak emission at 311-313 nm. The perception of pruritus severity was assessed using the Visual Analogue Scale (VAS) before starting the treatment and at each control. RESULTS: A complete remission of the cutaneous lesions and pruritus was documented in all patients after a median of 40.3 UV treatments and a median cumulative dose of 51.4 J/cm(2), with a lasting remission over a 6-month follow-up. The median VAS score at the beginning of the treatment was 86.6 (SD=6.64), whereas it decreased to 6.66 (SD=3.75) after 3 months of therapy. CONCLUSIONS: Our work provides evidence that NB-UVB phototherapy is useful for the treatment of the cutaneous symptoms and pruritus in ISM.

Minimal incidence of neonatal/infancy onset diabetes in Italy is 1:90,000 live births.

Until early 2000, permanent and transient neonatal diabetes mellitus (NDM), defined as diabetes with onset within 6 weeks from birth that requires insulin therapy for at least 2 weeks, were considered exceedingly rare conditions, with a global incidence of 1:500,000-1:400,000 live births. The new definition of NDM recently adopted, that includes patients with diabetes onset within 6 months of age, has prompted studies that have set the incidence of the permanent form alone between 1:210,000 and 1:260,000 live births. Aim of the present work was to ascertain the incidence of NDM (i.e. permanent + transient form) in Italy for years 2005-2010. Patients referred to the Italian reference laboratory for NDM between years 2005 and 2010 and screened for mutations in common NDM genes (KCNJ11, ABCC8, and INS) and for uniparental isodisomy of chromosome 6 (UDP6) were reviewed. A questionnaire aimed at identifying NDM cases investigated in other laboratories was sent to 54 Italian reference centers for pediatric diabetes. Twenty-seven patients with NDM born between 2005 and 2010 were referred to the reference laboratory. In this group, a mutation of either KCNJ11, ABCC8 or INS was found in 18 patients, and a case with UDP6 was identified. Questionnaires revealed 4 additional cases with transient neonatal diabetes due to UDP6. Incidence of NDM was calculated at 1:90,000 (CI: 1:63,000-1:132,000) live births. Thus, with the definition currently in use, about 6 new cases with NDM are expected to be born in Italy each year.

Characterization of the probiotic strain Vagococcus fluvialis in the protection of European sea bass (Dicentrarchus labrax) against vibriosis by Vibrio anguillarum.

Aquaculture is one of the main sources of income in many countries worldwide. Intensive farms are often affected by different infectious diseases that can decrease their final production. To control this situation, several antibiotics are frequently used with known environmental consequences. The aim of this study was to analyze different bacterial strains isolated from of gilthead sea bream, sea bass, sole and meagre guts, for use as probiotics in aquaculture. The strains were evaluated in vitro through various mechanisms of selection, such as the production of antagonistic effects against pathogens, production of antibacterial substance, adhesion to the intestinal mucus, competition for nutrients or binding site, and growth in intestinal mucus. A total of 50 bacterial strains were analyzed and only one showed excellent in vitro results for consideration as a candidate to be analyzed in vivo. The strain, identified as Vagococcus fluvialis, showed good protection against Vibrio anguillarum 975-1 in vivo in the experimental challenge, showing a relative percent survival of 42.3% higher than positive control group. Therefore, in conclusion we consider this strain to be a good candidate for use as a future probiotic in aquaculture.

Prevalence, severity and clinical features of psoriasis in fingernails and toenails in adult patients: Italian experience.

BACKGROUND: Psoriasis is a chronic inflammatory skin disease affecting 2.0-6.5% of the European population. Although the most striking clinical features of psoriasis involve the skin, other organs including nails and joints may be affected in a substantial proportion of patients. Literature reports nail involvement in 10-56% of psoriatic patients, with common physical and social impairment. However, the precise prevalence of specific clinical features of nail psoriasis is somewhat under-reported. OBJECTIVES: Our cross-sectional study aimed at describing the prevalence and the clinical features of nail involvement in adult psoriatic patients in a psoriasis referral centre in northern Italy. METHODS: A total of 178 (124 men, 54 women) consecutive adult patients (≥18 years old) with psoriasis were included. Psoriasis Area and Severity Index (PASI) and Nail Psoriasis Severity Index (NAPSI) scores were calculated for each patient. Relevant medical history was recorded. RESULTS: Nail involvement was present in 137 (99 men, 38 women) patients (76.9%). The most common nail abnormality was onycholysis, followed by crumbling, subungual hyperkeratosis, pitting and discoloration. Pitting and onycholysis were the most prevalent patterns observed in fingernails, whereas onycholysis and crumbling were the most frequent changes detected in toenails. The most frequently and severely affected nails were the fourth fingernail and the first toenail. The average PASI score was higher in individuals with nail involvement (12.0 vs. 8.7, P = 0.06). Nail changes were present in 85.7% of patients with psoriatic arthritis. CONCLUSIONS: Our study confirms that nail involvement may be overlooked in psoriasis patients. Different psoriatic patterns in the nail affect specific digits more frequently.

Permanent diabetes during the first year of life: multiple gene screening in 54 patients.

AIMS/HYPOTHESIS: The aim of this study was to investigate the genetic aetiology of permanent diabetes mellitus with onset in the first 12 months of age. METHODS: We studied 46 probands with permanent, insulin-requiring diabetes with onset within the first 6 months of life (permanent neonatal diabetes mellitus [PNDM]/monogenic diabetes of infancy [MDI]) (group 1) and eight participants with diabetes diagnosed between 7 and 12 months of age (group 2). KCNJ11, INS and ABCC8 genes were sequentially sequenced in all patients. For those who were negative in the initial screening, we examined ERN1, CHGA, CHGB and NKX6-1 genes and, in selected probands, CACNA1C, GCK, FOXP3, NEUROG3 and CDK4. The incidence rate for PNDM/MDI was calculated using a database of Italian patients collected from 1995 to 2009. RESULTS: In group 1 we found mutations in KCNJ11, INS and ABCC8 genes in 23 (50%), 9 (19.5%) and 4 (8.6%) patients respectively, and a single homozygous mutation in GCK (2.1%). In group 2, we identified one incidence of a KCNJ11 mutation. No genetic defects were detected in other loci. The incidence rate of PNDM/MDI in Italy is estimated to be 1:210,287. CONCLUSIONS/INTERPRETATION: Genetic mutations were identified in ~75% of non-consanguineous probands with PNDM/MDI, using sequential screening of KCNJ11, INS and ABCC8 genes in infants diagnosed within the first 6 months of age. This percentage decreased to 12% in those with diabetes diagnosed between 7 and 12 months. Patients belonging to the latter group may either carry mutations in genes different from those commonly found in PNDM/MDI or have developed an early-onset form of autoimmune diabetes.

Thrombotic thrombocytopenic purpura mimicking an acute meningoencephalitis.

Thrombotic thrombocytopenic purpura is a rare, threatening disease characterized by thrombocytopenia, microangiopathic haemolytic anaemia and organ dysfunction, e.g., neurological impairment and renal insufficiency. We describe a patient with neurological impairment mimicking a meningoencephalitis in whom a thorough clinical evaluation along with appropriate laboratory tests led us to identify an underlying thrombotic thrombocytopenic purpura. The successful outcome of this patient was based on plasma exchange and immunosuppressive treatment. Thrombotic thrombocytopenic purpura should be considered in the differential diagnosis of patients presenting with any neurological abnormalities, anaemia and unexplained thrombocytopenia.

Homocysteine, vitamin B12 and folic acid levels in psoriatic patients and correlation with disease severity.

Hyperhomocysteinaemia represents an independent risk factor for atherosclerotic cardiovascular disease, stroke, peripheral arterial occlusive disease and venous thrombosis. Psoriasis is a chronic inflammatory skin disease associated with increased atherothrombosis and cardiovascular risk profile. The aim of this study is to investigate homocysteine, folic acid and vitamin B12 levels in a cohort of psoriatic patients and its relationship with the severity of the disease. A retrospective observational study in 98 patients with chronic plaque psoriasis and 98 healthy controls was performed. Total plasma homocysteine level, folic acid, vitamin B12 and PASI index were assessed in every patient. Patients with psoriasis had plasma homocysteine levels higher than controls (57% of cases and 25% of controls; p<0.0001). Folic acid and vitamin B12 plasma levels were lower in psoriatic patients than in controls (p = NS), lower levels of vitamin B12 were found in patients with hyperhomocysteinaemia compared to patients with a normal value of homocysteine (p = 0.0009). The severity of psoriasis assessed according to PASI (19.51+/-16.26) did not directly correlate either with higher levels of homocysteine or with vitamin B12 and folic acid plasma levels. In conclusion, a significantly higher prevalence of hyperhomocysteinaemia was found in psoriatic patients compared to healthy controls. A significant correlation between hyperhomocysteinaemia and lower vitamin B12 levels, but not folic acid, was evidenced. On the contrary, our data do not correlate the high level of homocysteine with higher PASI scores or psoriasis type, suggesting that homocysteine level can be considered an independent risk factor in psoriatic patients.

Narrowband ultraviolet B phototherapy in the treatment of cutaneous graft-versus-host disease in oncohaematological paediatric patients.

BACKGROUND: Graft-versus-host disease (GVHD) represents an important complication following allogeneic bone marrow transplantation. In recent years, narrowband ultraviolet B (NB-UVB, 311-313 nm) has been found to be a beneficial adjuvant treatment in patients refractory to first-line immunosuppressive drugs. OBJECTIVES: The aim of this study is to analyse retrospectively the clinical outcome of 10 GVHD paediatric patients treated with NB-UVB therapy. PATIENTS AND METHODS: Ten paediatric patients (six girls and four boys: median age 12.5 years, range 4-20) with cutaneous GVHD were enrolled in the study: five patients with chronic GVHD and five patients with an overlap syndrome GVHD. All patients had already been shown to be resistant to first-choice immunosuppressive protocols, and were treated with NB-UVB phototherapy until a clinical remission of skin lesions occurred. RESULTS: A complete response (absence of lesions) was achieved in 80% of the cases (eight patients) after a median number of 29 treatments, corresponding to a median of 7.5 weeks (52 days) of treatment (range 3-13 weeks), with an average cumulative dose of 28.71 J cm(-2) (range 1.02-70.38 J cm(-2)). Only two patients reported a partial remission (< 18% of body surface area involved). During the follow-up period, a complete remission after 1 year was observed in 75% of patients and after 2 years in 71% of the evaluable patients. CONCLUSIONS: This study provides evidence that NB-UVB phototherapy represents a valid second-line treatment in paediatric patients affected by GVHD and refractory to immunosuppressive first-line treatment.

Invasion and survival of Photobacterium damselae subsp. piscicida in non-phagocytic cells of gilthead sea bream, Sparus aurata L.

Fluorescence microscopy and gentamicin protection assays were used to investigate the ability of four Photobacterium damselae subsp. pisicida (Phdp) strains to adhere to and to invade the fish epithelial cell line, SAF-1, derived from Sparus aurata. All strains tested were detected intracellularly using both techniques, although internalization levels varied among strains. Treatment with cytochalasin D and experiments carried out at 4 degrees C demonstrated that a functional host cell cytoskeleton and active cell metabolism are necessary for bacterial internalization. Intracellular bacteria were detected for up to 7 days with a round morphology and were stained with DAPI, indicating that some bacterial cells may remain viable inside SAF-1 cells. Our in vitro findings indicate that Phdp are capable of adhering, entering and surviving within the non-phagocytic epithelial cell line SAF-1, which may be important for persistence and establishment of a carrier state in S. aurata.