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Background: The Glasgow prognostic score (GPS) reflects the host’s systemic inflammatory response and is a validated prognostic factor in lung cancer. However, little is known about the prognostic role in non-small cell lung cancer (NSCLC) patients treated with immunotherapy after platinum-based cytotoxic chemotherapy.Methods: This study used a lung cancer cohort of the Catholic Medical Center of Korea between January 2018 and September 2020. We included patients who were diagnosed with unresectable advanced stage NSCLC or recurrent disease after pulmonary resection and had received at least one regimen of platinum-based chemotherapy before the administration of immunotherapy. The prognostic value of the GPS was assessed in patients with NSCLC treated with anti-PD1 or anti-PD-L1 (pembrolizumab, nivolumab, or atezolizumab). The GPS was calculated using C-reactive protein and albumin concentrations within one week before starting anti-PD1 or anti-PD-L1 treatment. Results: A total of 78 patients with NSCLC treated with immunotherapy as second or further-line therapy after platinum-based chemotherapy were included in the study. Kaplan-Meier analysis revealed that higher GPS values were significant predictors of shorter immune-related progression-free survival (irPFS) (log-rank P < 0.001) and overall survival (OS) (log-rank P < 0.001). In the Cox regression multivariate analysis, the hazard ratios for irPFS were 0.249 (95% confidence interval [CI]: 0.084 – 0.739, P = 0.012) for PD-L1 expression ≥ 50% and 9.73 (95% CI: 2.931 – 32.298, P < 0.001) for a GPS of 2 relative to a GPS of 0. Older age (P = 0.033), lower PD-L1 expression (P = 0.036), and higher GPS values (P = 0.007) were independently associated with shorter OS.Conclusions: Higher GPS values were identified as a poor prognostic factor for OS and irPFS in NSCLC patients who received immunotherapy as second or further-line therapy after platinum-based chemotherapy.

BACKGROUND: Anemia is a frequent finding in cancer patients. Pre-treatment anemia is known to be associated with poor survival after surgery or stereotactic body radiation therapy of non-small cell lung cancer (NSCLC). However, little study was conducted in NSCLC with activating epidermal growth factor receptor (EGFR) mutations. METHODS: This was a multicenter retrospective study conducted in seven university teaching hospitals in the Republic of Korea from January 2009 to February 2016. A total of 290 patients were diagnosed with NSCLC harboring sensitizing EGFR mutations and treated with EGFR-tyrosine kinase inhibitor (TKI) as 1st line. Of these patients, 104 met the exclusion criteria. Pre-treatment anemia was defined according to World Health Organization criteria (Hb concentration

A higher platelet-to-lymphocyte ratio (PLR) has a clinical correlation with shorter survival in non-small cell lung cancer (NSCLC). The present study evaluated the association between the PLR and survival in patients with advanced NSCLC with malignant pleural effusion (MPE). Between January 2012 and July 2016, 237 patients with stage IV NSCLC were selected for evaluation. Receiver operating characteristic analysis was used to determine a cutoff for the PLR. Clinicopathological characteristics were compared between the high and low PLR groups, and the role of PLR as a predictive/prognostic maker was investigated. Among the 237 patients, 122 were assigned to the low PLR group and the other 115 to the high PLR group. According to multivariate analysis, male sex, not receiving active anticancer treatment, low hemoglobin level, low albumin level, high C-reactive protein level, and high PLR were identified as significant risk factors for shorter overall survival (OS) (p = 0.010, p = 0.047

Brassicaceae consists of important species that have significant amounts of metabolites, and many studies have been carried out in order to understand the mechanism that improves the content of these metabolites. In Brassicacea, Cardamine manshurica Nakai is one of the important edible plants and is rich in oil, fiber, and various nutrients. In this study, we constructed cDNA library using leaves from 4 week-old plants and analyzed the ESTs of C. manshurica Nakai. One thousand thirty-nine ESTs were discovered which assembled to form 468 unigenes. The latter contained 116 contigs and 352 singletons. Similarity search of these ESTs with BLASTX revealed similarities with Arabidopsis thaliana 285 (31.9%), Arabidopsis lyrata 172 (19.3%), Capsella rubella 162 (18.1%), and Eutrema salsugineum 137 (15.3%). ESTs were functionally categorized into molecular function, biological process, and cellular component, and each category took 10.6%, 58.5%, and 30.9%, respectively. The functional analysis also found that 94.9% of ESTs showed at least one GO ID. Microsatellite analysis of 468 unigene sequences revealed 225 structures of which Di-, Tri-, Tetra-, Penta-repeats were 35.6% (80/225), 63.1% (142/225), 0.9% (2/225), and 0.4% (1/225), respectively. The results from our study can be a valuable resource for Cardamine research.

The multifunctional enzyme transglutaminase 2 (TG2) primarily catalyzes cross-linking reactions of proteins via (γ-glutamyl) lysine bonds. Several recent findings indicate that altered regulation of intracellular TG2 levels affects renal cancer. Elevated TG2 expression is observed in renal cancer. However, the molecular mechanism underlying TG2 degradation is not completely understood. Carboxyl-terminus of Hsp70-interacting protein (CHIP) functions as an ubiquitin E3 ligase. Previous studies reveal that CHIP deficiency mice displayed a reduced life span with accelerated aging in kidney tissues. Here we show that CHIP promotes polyubiquitination of TG2 and its subsequent proteasomal degradation. In addition, TG2 upregulation contributes to enhanced kidney tumorigenesis. Furthermore, CHIP-mediated TG2 downregulation is critical for the suppression of kidney tumor growth and angiogenesis. Notably, our findings are further supported by decreased CHIP expression in human renal cancer tissues and renal cancer cells. The present work reveals that CHIP-mediated TG2 ubiquitination and proteasomal degradation represent a novel regulatory mechanism that controls intracellular TG2 levels. Alterations in this pathway result in TG2 hyperexpression and consequently contribute to renal cancer.

Histone acetylation has a central role in establishing an active chromatin environment. The functional contribution of histone acetylation to chromatin transcription is accomplished by a dominant action of histone acetyltransferases over repressive histone-modifying activities at gene promoters; misregulation of these dynamic events can lead to various diseases. Here, we describe the synthesis and characterization of transducible peptides derived from histone H4 N-terminal tail as a molecular tool to establish and maintain the active state of p53 target genes. Cellular experiments demonstrate a distinct increase in p53 transactivation by acetylated H4 tail peptides, but only a modest change by unmodified H4 tail peptides. The molecular basis underlying the observed effects involves the selective interaction of the tail peptides with G9a histone methyltransferase and histone deacetylase 1 (HDAC1) and the disruption of their occupancy at p53 target promoters. Furthermore, treatment of xenograft models and cancer cell lines with the tail peptides sharply decline tumor cell growth and enhances apoptosis in response to DNA damage. These results indicate that H4 tail peptide mimics upregulate p53 transcription pathway and may be used as a novel strategy for anticancer therapy.

The effect of different solvent-based (methanol, ethanol, and aqueous) extracts of Liriope platyphylla at 0, 0.1, 1.0, and 2.0 mg doses on hematology and innate immune response in olive flounder, Paralichthys olivaceus, against Flexibacter maritimus is reported. The red blood cells and white blood cells significantly increased with ethanol and aqueous extracts with any dose against F. maritimus while the hematocrit level significantly increased with methanol or ethanol extracts. However, the hemoglobin content significantly increased irrespective of the doses and type of extracts, whereas the mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration significantly decreased. The lymphocytes and monocytes significantly increased in all doses and extracts, whereas neutrophils increased in ethanol or aqueous extracts. However, the thrombocytes level neither the doses nor extracts did not significant changes to the control against pathogen. Total protein and globulin levels significantly increased with all the doses of ethanol extract while does not significant change in methanol extract. The phagocytic activity and complement activity were significantly enhanced with any dose of ethanol extract while with aqueous extract it was only at 1.0 and 2.0 mg doses. At all the doses of each extract significantly enhanced the leucocytes activity against F. maritimus. The results confirm that the ethanolic and aqueous extracts of L. platyphylla stimulate the immune response against F. maritimus in P. olivaceus.

The protective efficacy of liposomes-entrapped with whole cell bacterial vaccine (WCV) on immune response and protection against Vibrio harveyi infection in Epinephelus bruneus was investigated at weeks 1, 2, and 4. The cumulative mortality was 25% in WCV immunized fish whereas it was 35% and 40% with activated whole cell V. harveyi alone (WC) or liposome alone against pathogen; the mortality was 90% in non-immunization fish against pathogen. Total protein significantly increased in WCV and WC groups on weeks 2 and 4. The albumin, globulin, and albumin:globulin ratio significantly increased in WCV and WC groups on week 4. The antibody titres significantly increased in fish immunized with liposomes-entrapped WCV from weeks 1 to 4 where as in WC group only on week 4. The phagocytic activity significantly enhanced in liposomes-entrapped WCV group on weeks 1 and 2 and all other immunized groups on week 4. The complement activity was significantly enhanced in liposomes-entrapped WCV and WC immunized groups on weeks 2 and 4. However, the lysozyme activity significantly was enhanced in all immunization groups from weeks 1 to 4. These results suggest that fish immunized with liposomes-entrapped WCV and WC alone have the potential to modulate the immune system affording increased protection against V. harveyi infection in kelp grouper.

Philasterides dicentrarchi is a histophagous scuticociliate infecting Korean olive flounder farms in Jeju Island, South Korea, where it causes significant economic losses. However, the route of entry of these parasites in olive flounder is currently unknown. In the present study, we attempted experimental infection with different doses (concentrations) of P. dicentrarchi by intraperitoneal (IP) injection; maximum cumulative mortality of 95% was recorded on the administration of 2.1 × 108 ciliates/ml. In small size group (3 cm length, 24 ± 2 g wt) the ciliates were found in connective tissues of muscle, fins, and nervous tissue while in large size group (5 cm length, 47 ± 3 g wt), the ciliates were predominantly occurred with associated symptoms of liquefaction in the central nervous system. The major clinico-pathological manifestations were loss of scales, appearance of bleached spots that coalesced to form brownish patches, hemorrhagic, and severe dermal necrotic lesion. In addition there was degeneration of muscle fibers, necrosis of the hepatic parenchyma, and severe edema of the intestinal walls, extensive fouling, necrotic degeneration and hyperplasia in the gill bronchial epithelium. In this study, many ciliates were found in the blood vessel, brain, dorsal fins, muscle, kidney, and vertebra of infected flounder. The histopathological investigations are of significant importance in view of possible routes of entry into the host and doses of ciliates that trigger fast infection for potential utility of drugs as a strategy for the control of P. dicentrarchi in farmed olive flounder.

The present study determined the protection afforded by dietary administration with Pueraria thunbergiana extract in terms of mortality and innate immune response to Vibrio harveyi infection in kelp grouper, Epinephelus bruneus. The grouper was fed with 0% (control), 0.1%, 1.0%, and 2.0% supplementation diets at week 1, 2, and 4. All enriched diets significantly increased the weight gain (WG) from weeks 1 to 4 while specific growth rate (SGR), feed conversion ratio (FCR), and protein efficiency ratio (PER) did not change significantly when compared to the control. The white and red blood cells count, haemoglobin, haematocrite, lymphocytes, and monocytes significantly increased when fed with 1.0% and 2.0% supplementation diets against pathogen. Total protein content and albumin level significantly increased with 1.0% diet than when fed with 0.01% or 2.0% enriched diet. All the supplementation diets significantly enhanced phagocytic ratio and lysozyme activity from week 1 to 4 against pathogen. The phagocytic index, superoxide anion production, bactericidal activity, and anti-protease activity significantly increased in fish fed with 1.0% and 2.0% doses diets from weeks 1 to 4 as compared to the control, but not with 0.1% diet. The highest protection by means of lower cumulative mortality was recorded with 1.0% and 2.0% diets when compared to 0% and 0.1% diet. This study suggested that 1.0% and 2.0% supplementation diet significantly influence the growth, hematology, and enhance the innate immune system in kelp grouper, E. bruneus against V. harveyi.

The present study investigated the effect of Eriobotrya japonica extracts at 0%, 0.1%, 1.0%, and 2.0% doses supplementation with feed on non-specific immune response, hematological and biochemical profile, and disease resistance against Vibrio carchariae in kelp grouper Epinephelus bruneus at weeks 1, 2, and 4. The white blood cell (WBC) significantly increased in fish fed with 0.1%, 1.0%, and 2.0% diets on weeks 1 and 2 when compared to the control. However, the glucose always decreased from the control except on week 2 against pathogen. The serum total protein, albumin, and globulin significantly increased at week 2 but they did not changed significantly at weeks 1 and 4. The superoxide anion, lymphokines production index, and phogocytosis did not significantly increased in any diet on the first week whereas it was significantly enhanced in 1.0% and 2.0% supplementation diets on weeks 2 and 4 against V. carchariae when compared to control. All diets significantly enhanced the serum lysozyme activity, bactericidal activity, and haemolytic complement activity from weeks 1–4 as compared to control. The serum agglutinating antibody titre did not significantly enhance on the first week whereas it was significantly enhanced on weeks 2 and 4. Fish fed with 1.0% and 2.0% doses diets was found lower mortality than 0.1% diet. Thus, this study suggested that 1.0% and 2.0% doses supplementation diets could be advocated to enhance the immune response and production disease from V. carchariae in E. bruneus.

This study, reports the identification and analysis of ferritin M chain, from kelp grouper, Epinephelus bruneus (EbFerM); it comprises 1004 base pair (bp), including 528 bp open reading frame (ORF) which encodes 176 amino acid (aa) residues; the calculated molecular weight is 20 kDa. The 5′-untranslated region (UTR) possesses 476 bp proceeded by a putative Iron Regulatory Element (IRE). Pair wise alignments showed that EbFerM shared 94% identity with that of Larimichthys crocea and Sciaenops. It is expressed in abundance in liver, spleen, and kidney when challenged with Vibrio anguillarum, lipopolysaccharide (LPS), or poly I:C.

Feeding kelp grouper, Epinephelus bruneus (26.1 ± 1.4), with 0%, 0.1%, 1.0%, and 2.0% Kalopanax pictus extract-supplementation diets, for 30 days reduced mortality on being challenged intraperitoneally 100 μl with bacterium Vibrio alginolyticus (2.1 × 10 7 cfu ml −1 ) and ciliate parasite Philasterides dicentrarchi (2.3 × 10 7 ciliates ml −1 ). The red blood cells (RBC), white blood cells (WBC), haemoglobin, haematocrit, lymphocytes, and monocytes levels significantly increased in kelp grouper fed with all doses of K. pictus -supplementation diets and challenged with bacterium and parasite when compared to control. However, the levels of mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), neutrophils, and thrombocytes did not significantly. The phagocytic activity, complement activity, and antiprotease activity did not significantly change in kelp grouper fed with 0.1% K. pictus -supplementation diets and challenged with bacterium and parasite. The respiratory activity, lysozyme activity, bactericidal activity, total protein level, and myeloperoxidase levels significantly increased in kelp grouper fed with all the doses of K. pictus -supplementation diet and challenged with bacterium and parasite. However, α2-macroglobulin level significantly increased with 1.0% diet, but not with 0.1% and 2.0% diets. Therefore this study suggests that 1.0% and 2.0% K. pictus -supplementation diets positively protected and enhanced the immune system in kelp grouper E. bruneus against V. alginolyticus and P. dicentrarchi infection.

Kelp grouper, Epinephelus bruneus, fed for 30 days with 0% (control), 0.1%, 1.0%, and 2.0% of Styrax japonica supplementation diets, led to reductions in mortality after being challenged with a bacterium (Vibrio harveyi) and a ciliate protozoan (Uronema marinum). The enriched diets significantly increased the survival rate as compared to the controls. The phagocytic and respiratory activities were significantly increased in kelp groupers given 1.0% and 2.0% enriched diets. The complement activity, lysozyme activity, serum bactericidal activity, and total protein level significantly increased with any enriched diet against the pathogens; however antiprotease activity and myeloperoxidase levels significantly increased only with 1.0% and 2.0% enriched diets while the α2-macroglobulin level was significantly enhanced with 1.0% enriched diet. The study suggests that incorporation of S. japonica at 1.0% and 2.0% level in the diet significantly enhances the immune responses in the kelp grouper E. bruneus against V. harveyi and U. marinum.

The present study was reported the effect of probiotics and herbals mixture supplementation diet on growth, blood constituents, and nonspecific immune response in olive flounder Paralichthys olivaceus against Streptococcus parauberis on weeks 1, 2, 4, 6, 8, 10, and 12 after injected intraperitoneally (i.p.) with 50 μl of PBS (phosphate buffer saline) containing S. parauberis (2.1 × 10⁷ CFU ml⁻¹). The initial weight did not significantly increased in supplementation diet group from 1 to 4 weeks, whereas it was significantly increased from weeks 6 to 12 as compared to fish fed without supplementation diet. The serum aspartate aminotransferase (SGOT) and alanine aminotransferase (SGPT) activities significantly increased from weeks 4 to 12 in infected fish fed with supplementation diet compared to fish fed without supplementation diet. However, the total protein (TP) and glucose (GLU) levels were significantly increased in infected fish fed with supplementation diet after 6 weeks. The phagocytic, respiratory burst, complement, and lysozyme activities significantly enhanced in infected fish fed with supplementation diet from weeks 4 to 12 as compared to fish fed without supplementation diet. These results suggested that different probiotics and herbals mixture supplementation diet enhanced the growth, blood biochemical constituents, and nonspecific immunity in olive flounder against S. parauberis.

The present study evaluated the efficacy of dietary doses of Lactuca indica extract on immunological parameters and disease resistance against Streptococcus iniae infection in kelp grouper, Epinephelus bruneus . Fishes were fed with L. indica enriched diet at 1.0% and 2.0%; both the diets significantly enhanced NBT level and phagocytic activity on week 2 and 4 compared to control diet whereas the changes did not manifest on first week. The total immunoglobulin significantly enhanced in fish fed with 1.0% and 2.0% enriched diets whereas the lysozyme activity significantly enhanced all the diets from weeks 1 to 4. Further the enriched diets at 1% and 2% level resulted in lower mortality of 45% and 40% respectively indicating higher protection from S. iniae infection than 0.1% diet that resulted in 55% mortality. The results suggest that the dietary supplementation of L. indica extract stimulates the immunological parameters and increases disease resistance in E. bruneus against S. iniae infection.

The effect of traditional Prunella vulgaris extracts enriched diets at 0%, 0.01%, 0.1%, and 1.0% doses were investigated the non-specific immune response and disease resistance of olive flounder (Paralichthys olivaceus) against Uronema marinum on weeks 1, 2, and 4. The scuticocidal activity significantly increased in groups fed with 0.1% and 1.0% doses of P. vulgaris enriched diets against U. marinum on weeks 2 and 4. The effect of P. vulgaris extract enriched diets on respiratory burst activities significantly increased on weeks 2 and 4 in the 0.1% and 1.0% doses. On the other hand, the phagocytic activities significantly enhanced at 0.1% and 1.0% doses of P. vulgaris extract enriched diet from weeks 1 to 4. All diets a significantly increase in plasma lysozyme activity on weeks 2 and 4. However, complement activity was significant in the 0.1% and 1.0% doses of P. vulgaris on fourth week. Feeding with 0.1% and 1.0% diets was the most effective and reduced the mortality then 0.01% diet. These results indicate that 0.1% and 1.0% doses of P. vulgaris extracts enriched diets potentially enhanced the non-specific immune response and disease resistance of P. olivaceus against U. marinum.

Monkey head mushroom, Hericium erinaceum enriched diet fed to olive flounder Paralichthys olivaceus infected with Philasterides dicentrarchi afforded protection by enhancing the innate immune response. In the infected fish the scuticocidal activity, respiratory burst activity, and phagocytic activity did not significantly change at any time when fed with 0.01% H. erinaceum. However, on being fed with 0.1% and 1.0% enriched diets the chosen immune parameters significantly increased from week 1 to 4 when fish and challenged with parasites. The lysozyme activity significantly increased more from weeks 1 to 4 in the infected group than in the control. The cumulative mortality was 90% in control group whereas it was 45% and 30% when fed with 0.1% and 1.0% enriched diets respectively. The present study suggests that feeding with 0.1% and 1.0% enriched diets enhanced the immune parameters and affords protection from P. dicentrarchi infection in olive flounder.

The effect of five probiotics, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus sakei, Bacillus subtilis, and Saccharomyces cerevisiae as individual and mixed enriched diet on the seasonal prevalence, activity and intensity of Uronema marinum infection in olive flounder Paralichthys olivaceus is reported. The growth performance, feed efficiency, blood biochemistry, survival rate, and non-specific immune response of U. marinum infected olive flounder on week 0, 1, 2, 4, 6, and 8 were quantified. The prevalence and infection intensity reached a peak from June to December and then it declined from December to March. The scuticocidal activity in the serum was significantly higher when fed with L. plantarum, L. acidophilus, and S. cerevisiae diets on weeks 2-8. All enriched diets significantly enhanced the weight gain significantly between week 6 and 8; the feed efficiency registered a significantly increase from week 4 to 8 when compared to infected fish fed with control diet. Infected fish fed with L. plantarum-supplemented diet had higher survival rate than with other enriched diets. The serum aspartate aminotransferase (GOT) and alanine aminotransferase (GPT) levels significantly increased when fed with L. plantarum, L. acidophilus or S. cerevisiae-supplemented diet. Total protein (TP) and glucose (GLU) level significantly increased with any enriched diet from week 4 to 8. The superoxide anion production and serum lysozyme activity registered a significant increase when fed with L. plantarum, L. acidophilus, and S. cerevisiae-supplemented diet from week 4-8. The present study concludes that L. plantarum, L. acidophilus, and S. cerevisiae-supplemented diets act as immunostimulants enhancing the growth, feed efficiency, blood biochemistry, survival rate, and non-specific immune response in U. marinum infected olive flounder.

In olive flounder, Paralichthys olivaceus infected with lymphocystis disease virus (LDV) bath treatment with formalin, hydrogen peroxide or Jenoclean at 50, 100, and 200 ppm daily 10 min for 60 days enhanced the innate immune response and disease resistance. Jenoclean enhanced the immune parameters at the lowest concentration of 50 ppm; on the other hand, hydrogen peroxide bath treatment enhanced the immunity level at 100 ppm, while formalin was effective only at 200 ppm. A low cumulative mortality and high relative percent survival was noted in Jenoclean treated group followed by formalin and hydrogen peroxide treated groups. In olive flounder at low concentrations of 50 ppm or 100 ppm hydrogen peroxide and Jenoclean effectively prevent LDV infection.

We report the growth, innate immune response, and disease resistance in olive flounder (Paralichthys olivaceus) challenged with Streptococcus iniae after feeding with diet enriched with Zooshikella sp. strain JE-34 three different concentration i.e. Low (3.4 x 10(4), n = 50), medium (3.5 x 10(6), n = 50), and high (3.4 x 10(8), n = 50) cfu ml(-1) supplemented diets, the changes were monitored on weeks 1, 2, 4, 8, 12, and 16. With all diets the innate immune parameters, such as superoxide anion production, phagocytic and lysozyme activity were not enhanced on week 1 and 4. On the other hand, all tested immune parameters in the treated groups significantly enhanced after 8th week; the weight gain significantly increased after 4th week in fish fed with enriched diets. The mortality in olive flounder after administration with high concentration diet showed 25%. With low and medium enriched diets the mortality was 40% and 35%, respectively. In the infected untreated group mortality was 85% while there was no mortality in the control group. The results suggested that Zooshikella sp. strain JE-34 enriched diets could be used to enhance the innate immune response and disease resistance of P. olivaceus against S. iniae.

Etiological investigations have often implicated Aeromonas hydrophila, a heterotrophic Gram-negative pathogenic aquatic bacterium, in a variety of systemic and localized fish diseases. The aim of this study was to test the efficacy of a herbal concoction against A. hydrophila-infected goldfish (Carassius auratus). After the pathogen was intramuscularly injected, scales sloughed off on the site of administration, with the appearance of a muscular hemorrhagic protuberance that progressed into an extensive ulcerative dermatitis associated with focal hemorrhage, edema, and dermal necrosis exposing the underlying muscles. Progression of the disease affected the organs in the following order: muscle, gills, liver, and finally the heart tissue. The dip treatment with the concoction (1% herbal concoction dip treatment for 5 min/d daily at 1100 h) restored the histoarchitecture of the altered primary gill lamellae, liver, heart, and muscle. The recovery changes are reported.

Philasterides dicentrarchi is a histophagous scuticociliate causes fatal scuticociliatosis in farmed olive flounder Paralichthys olivaceus . The average monthly prevalence of scuticociliatosis due to P. dicentrarchi infections was increased from May to July (40 ± 3.1% to 79.4 ± 1.7%) and it decreased from August to November (63 ± 2.3% to 30 ± 2.6%) in olive flounder farms at Jeju Island, South Korea during 2000–2006. The prevalence of mixed infection along with Vibrio spp. bacterial infection was 49 ± 7.2% than that of other mixed infection. At present no effective control measure for P. dicentrarchi infection has been described and large production losses continue. In the present study, formalin, hydrogen peroxide and Jenoclean chemotheraputants were used for bath treatment. Among Jenoclean at a low concentration of 50 ppm proved effective. The results were confirmed with in vitro motility assessments and morphological changes scoring system in P. dicentrarchi . On the other hand, similar trend was noted following hydrogen peroxide treatment at this concentration, but formalin was only moderately effective. Either hydrogen peroxide or Jenoclean are the promising compounds effective at low concentrations with short application time for P. dicentrarchi . Therefore, these substances were evaluated on day 10, 20 and 30 for their ability to enhance innate immune response and disease resistance against P. dicentrarchi in olive flounder after chemotheraputants bath treatment with 100 ppm for 30 min per day. All the tested immune parameters were enhanced by treatment with Jenoclean, but not formalin and hydrogen peroxide. These findings suggest that Jenoclean bath treatment can be used for ensuring the heath of cultured marine fish against internal parasites such as P. dicentrarchi .

Goldfish ( Carassius auratus ) weighing 13 ± 2 g were administered intramuscularly a sublethal dose (1.8 × 10 3 cfu/ml) of Aeromonas hydrophila to induce ulcerative dermatitis. On day 3 and day 6 after infection the fish were dip-treated (for 5 min/day) with a tri-herbal concoction of Curcuma longa, Ocimum sanctum and Azadirachta indica (1%). The LD 50 value was recorded at a concentration of 2.3 × 10 4 between 30 and 36 days after infection. In the infected untreated group the cumulative mortality was higher, while in the early-treated group (day 3) there was no mortality. In the late-treated group (6th day) the mortality increased to 23.3% on day 36. In the infected group the size of ulcers progressively increased from 43.3% of the body length on day 18 to 86.7% on day 36. In the early-treated fish the size of ulcers was 23.3% of the body length on day 18; by day 36 after treatment the ulcer had completely healed. In contrast, there was only a moderate recovery in the late-treated group. From the results it can be concluded that early tri-herbal treatment ensures successful recovery from ulcerative dermatitis induced by A. hydrophila .

Recently, mass mortality due to histophagous scuticociliate, Philasterides dicentrarachi in olive flounder, Paralichthys olivaceus occurred in aquaculture farms of Jeju Island, South Korea. We have artificially induced infection through immersion on day 12, the density of ciliates slowly decreased in the experimental tanks; on day 28 to 2300cells/ml in 3cm group and 1800cells/ml in 5cm group. The survival rate of P. olivaceus on days 24 and 28 of infection in the 3cm group was decreased to 4 and 6, respectively; however, no mortality was recorded in the 5cm group. In the control group 100% survival in the chosen period of both groups occurred except in the 3cm group whereas on days 24 and 28 the death rate was 1 and 2, respectively. Many ciliates with ingested red blood cells were observed in the gills, skeletal muscle, skin, fins and brains of infected flounder. The parasites were also observed in the lamina propria of the digestive tract, pharynx, peritoneal fluid, cerebrospinal fluid and cornea. Histopathological and inflammatory changes were characterized by swelling and generalized erythema and muscle tissue necrosis. Affected fish showed varying levels of tissue damage including severe epidermal and dermal necrotic lesions. Loss of scales, appearance of bleached spots that coalesced to form brownish patches, haemorrhagic and severe dermal necrotic lesions were the major clinico-pathological manifestations. The parasite was also seen distributed extensively in the entire brain causing widespread nerve necrosis after a prolonged period of infection. These results appear to support the hypothesis that the P. dicentrarchi may be penetrating via the gills or the skin, and then travel via the bloodstream to other parts of the body, including the body cavity.

We have investigated the effects of mushroom mycelium (Phellinus linteus, Coriolus versicolor) mixed cultural extract on the immune responses of olive flounder, Paralichthys olivaceus. The mixed culture extracts were evaluated for the growth, hematology, lysozyme activity, leukocyte phagocytic activity, and disease resistance against Vibrio anguillarum. In the effect of the growth, the body weight and length gain in the group, which fed with mushroom mycelium mixed cultural extract, were 52 g and 3.4 cm higher than that in the control, respectively. For the hematology, the administration of mushroom mycelium mixed cultural extract resulted in increase of glucose. However, there was no distinct differences in GOT (glutamic oxaloacetic transaminase), GPT (glutamlc pyruvic transamlnase), TG, TP, and LDH (lactate dehydronase) among each group. The activities of lysozyme were 80% higher in the experimental groups than in the control. The activities of leucocyte were 66% higher in the experimental groups than in the control. Although lysozyme activity and leucocyte activity showed somewhat decrease after 12 weeks, these activities were still higher than in the control. The cumulative mortality (%) after an artificial challenge with of Vibrio anguillarum per fish was 25% higher in the experimental groups than the control.

Background: NADPH oxidase (NOX) plays an important role in inflammatory responses by producing reactive oxygen species. The inhibition of NOX has been shown to participate in anti-inflammatory action in some inflammatory models. The aim of this study was to investigate the effects of diphenyleneiodonium (DPI), a NOX inhibitor, on lipopolysaccharide (LPS)-induced acute lung injury (ALI) model. Methods: Sprague Dawley rats were intraperitoneally administered DPI (5 mg/kg) 30 minutes after intratracheal instillation with LPS (3 mg/kg). After 6 hours, bronchoalveolar lavage fluid (BALF) and lung tissue were collected. The numbers of inflammatory cells and cytokine release (TNF-α, IL-1β, and IL-6) in BALF were detected by cell counting and ELISA. In lung tissues, NF-κB, myeloperoxidase (MPO) activity and ALI score were measured. Inducible nitric oxide synthase (iNOS) was examined by immunohistochemistry. To identify the anti-inflammatory mechanism of DPI, activation of MAPK and NF-κB pathways was measured by western blot analysis. Results: DPI-treated rats showed significantly reduced numbers of inflammatory cells and production of inflammatory cytokines in BALF compared to LPS-treated rats. In lung tissues, NF-κB and MPO activity, ALI scores, and iNOS expression were significantly decreased in DPI-treated rats compared to LPS-treated rats. Western blot analysis demonstrated that DPI significantly suppressed LPS-induced activation of MAPK and NF-κB pathways. Conclusion: Our results suggest that DPI may have an anti-inflammatory effect on LPS-induced ALI via suppression of the activation of MAPK and NF-κB pathways. DPI could be a useful therapeutic agent of ALI treatment.

We report the effect of Broussonetia kazinoki as a feed additive on disease resistance in olive flounder Paralichthys olivaceus, against Streptococcus parauberis. The cumulative mortalities were lower in fish fed with diets supplemented with 1.0% and 2.0% than in those fed with 0% and 0.1% diets. Phagocytic, complement and lysozyme activities were significantly enhanced by 0.1%, 1.0% and 2.0% B. kazinoki diets. Respiratory burst activity was significantly enhanced by 1.0% and 2.0% diets. This study suggests that diets supplemented with 1.0% and 2.0% B. kazinoki enhance the immune parameters concerning resistance against S. parauberis infection in P. olivaceus.

We studied the effects of diet supplemented with 0% (control), 0.1%, 1.0% and 2.0% Zanthoxylum piperitum on the immune response and disease resistance in kelp grouper Epinephelus bruneus against Vibrio carchariae. The phagocytic and lysozyme activities were significantly enhanced in fish fed with diets supplemented with 1.0% and 2.0% Z. piperitum. The cumulative mortality of fish fed with 1.0% and 2.0% supplemented diets was lower than that of the controls. Therefore, it was suggested that diets supplemented with 1.0% and 2.0% Z. piperitum positively enhanced the innate immune system in E. bruneus against V. carchariae.

The effect of dietary Coriolus versicolor supplementation at 0%, 0.01%, 0.1%, and 1.0% on the hematology and innate immune response of olive flounder (Paralichthys olivaceus) against the ciliate Pseudocohnilembus persalinus was studied after one, two, and four weeks of feeding. White blood cells were significantly higher in fish fed the 0.1% and 1.0% diets than in those fed the control and 0.01% diets from week two onwards. Similarly, red blood cells were significantly higher in the 0.1% and 1.0% diets than the other diets at week 4. Hemoglobin was significantly highest in the 1.0% diet and hematocrit significantly highest in the 0.1% diet at weeks 2 and 4. Total protein, glucose, and calcium were significantly higher in fish fed the 0.1% and 1.0% supplementation diets from weeks 1 to 4. Scuticocidal and lysozyme activity were significantly enhanced in fish fed the 0.1% and 1.0% diets from week 1 to 4 while respiratory burst activity was significantly higher in fish fed the 0.1% and 1.0% diets from week 2 to 4. Cumulative mortality was lower in groups fed the 0.1% and 1.0% diets than in groups fed the control or 0.01% diets. The present study suggests that dietary supplementation with 0.1% or 1.0% C. versicolor protects hematological parameters, improves innate immune response, and affords protection from P. persalinus infection in olive flounder.

Background: The release of interferon-gamma (IFN-γ ) by T lymphocytes increases after rechallenge with Mycobacterium tuberculosis antigen, especially, at a localized site of tuberculosis (TB) infection. We aimed to compare the clincial efficacy of two commercial IFN-γ release assays from pleural fluid for the diagnosis in tuberculous pleurisy. Methods: We performed T-SPOT.TB and QuantiFERON-TB Gold tests simultaneously on pleural fluid and peripheral blood samples from patients with pleural effusion, in South Korea, an area with intermediate TB burden. Results: Thirty-six patients were enrolled prospectively, and tuberculous pleurisy was found in 21 patients. Both the numbers of IFN-γ secreting T cells and the concentration of IFN-γ were greater in the pleural tuberculous group, comparing with the non-tuberculous group. Moreover, in the tuberculous group, there was a significant difference in IFN-γ producing spot-forming cells using the T-SPOT.TB method between pleural fluid and peripheral blood. The receiver operating characteristic (ROC) curve, was the greatest for pleural fluid T-SPOT.TB test, followed by peripheral blood T-SPOT.TB test, peripheral blood QuantiFERON-TB Gold test, and pleural fluid QuantiFERONTB Gold test (area under the ROC curve of 0.956, 0.890, 0.743, and 0.721, respectively). The T-SPOT.TB assay produced less indeterminate results than did QuantiFERON-TB Gold assay in both pleural fluid and peripheral blood. Conclusion: These findings suggest that the pleural fluid T-SPOT.TB test could be the most useful test among the IFN-γ release assays for diagnosing tuberculous pleurisy in an area with an intermediate prevalence of TB infection.

The present study investigated the effect of 1.0% chitin and chitosan supplementation diets on haematology and immune response in Kelp grouper, Epinephelus bruneus against protozoan parasite, Philasterides dicentrarchi. The red blood cells (RBC), white blood cells (WBC), haemoglobin levels, lymphocytes, monocytes, and neutrophils significantly increased in kelp grouper fed with chitin or chitosan enriched diets against P. dicentrarchi. The mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), and thrombocytes did not significantly change against pathogen. The phagocytic activity, respiratory burst activity, complement activity, antiprotease activity, and α2-macroglobulin were significantly enhanced in fish fed with 1% chitin and chitosan diet on weeks 2 and 4. The lysozyme activity, total protein, and myeloperoxidase activity significantly increased in fish fed with chitin or chitosan supplementation diet from weeks 1 to 4 against pathogen. The cumulative mortality was found low in fish fed with chitin and chitosan enriched diets than those of control against pathogen. The present study suggests that supplementation of 1.0% chitin or chitosan in diets positively enhances immune response and affords disease resistance in kelp grouper, E. bruneus against P. dicentrarchi infection.

The effect of Coriolus versicolor extract supplemented diets on innate immune response and disease resistance in kelp grouper, Epinephelus bruneus against Listonella anguillarum , is reported. Kelp grouper were divided into four groups of 25 each and fed with C. versicolor enriched diets at 0% (control), 0.01%, 0.1%, and 1.0% level. After 30 days of feeding, all fish were injected interaperitoneally (i.p.) with 50 μl of L. anguillarum (4.7 × 10 7 CFU) to investigate the immune parameters at weeks 1, 2, and 4. The reactive oxygen species and reactive nitrogen species production were significantly enhanced in fish fed with 0.1% and 1.0% supplementation diets from weeks 1–4 when compared to the non enriched diet fed and infected control. The phagocytic activity significantly increased with 0.1% and 1.0% diets on weeks 2 and 4. The leucocyte myeloperoxidase content, lysozyme activity, and total protein level significantly increased when fed with 0.1% and 1.0% supplementation diets from weeks 1–4. The cumulative mortality was 35% and 45% in 1.0% and 0.1% enriched diet fed groups whereas it was 55% and 80% in 0.01% and 0% groups respectively. The present results suggest that diets enriched with C. versicolor at 0.1% or 1.0% level positively enhance the innate immune system and affords protection from L. anguillarum .

The kelp grouper, Epinephelus bruneus, is an economically important intensively cultured species in Southeast Asia. Despite the insatiable demand its large-scale production has been hindered by problems associated with water quality, nutrition, and diseases especially due to increased rearing density. It is generally accepted that in fish both innate and adaptive immune system provide protection from diseases. In the present study a cDNA library of Streptococcus iniae-challenged kelp grouper was constructed to identify the genes that reveal molecular mechanism, physiological functions, and gene expression in different tissues using expressed sequence tags (ESTs) and RT-PCR strategy. Of a total of 2170 ESTs examined 279 (12.9%) were identified as contig and 860 (39.6%) as singletons. A total of 190 important immune and enzyme related genes (16.7%) were identified in both contig and singletons. The key immune molecules identified comprise complement factors, chemotaxin, chemokine, Fas ligand, ferritins, hepcidin, lysozyme c, MHC, and TLR which are involved in the innate or adaptive immune system. Among the genes a full-length cDNA of leukocyte cell-derived chemotaxin-2 (EbLECT2) with 540 base pair (bp) was identified; it consists of a 5'-untranslated region (UTR) of 17 bp, a 3'-UTR of 76 bp, and a stop codon TAA in 3'-UTR. The EbLECT2 is an important molecule in the innate immunity. It is a multifunctional protein involved in cell growth, differentiation, and autoimmunity. The open reading frame (ORF) of the EbLECT2 encodes with 155 amino acid (aa) residues with a predicted molecular weight and isoelectric point (pI) of 17 kDa and 9, respectively. The close phylogenetic relationship of EbLECT2 shares the highest similarity with the already reported LECT2 from Epinephelus coioides (96%) and Epinephelus akaara (94%). EbLECT2 mRNA was expressed predominantly in liver, spleen, and kidney while the expression was moderate in gills, heart, and muscle in E. bruneus after being challenged with LPS from Escherichia coli and pathogenic bacterium Vibrio anguillarum both of which involve the immune defense system. Further, the recombinant mature EbLECT2 (rEbLECT2) was successfully expressed in E. coli BL21 (DE3), and the antiserum against EbLECT2 was obtained for further investigations. The significant number of ESTs genome results obtained constitutes a powerful resource for further investigation to establish the gene discovery, functional genomic research, molecular mechanisms, and development of microarrays for the gene expression studies in kelp grouper.

Olive flounder, Paralichythys olivaceus fed with pomegranate enriched diet and challenged with or without Philasterides dicentrarchi had a significantly higher white blood cell (WBC) count on weeks 2 and 4 than the infected group fed with non enriched diet (standard diet). Similarly the red blood cell (RBC) counts did not significantly change in control and treated fish on weeks 1 and 2. It was significantly increased in treated fish on week 2 when compared to the control. In both the groups the hemoglobin (Hb) and hematocrit (Ht) levels significantly increased on weeks 2 and 4. The mean corpuscular volume (MCV) did not significantly change at any time in both groups whereas mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) increased significantly on week 4 in the treated group. The leukocytes such as lymphocytes (Lym), monocytes (Mon), neutrophils (Neu), and biochemical parameters such as total protein (TP), glucose (GLU), and calcium (CAL) levels significantly increased in treated groups on week 2 or 4 as compared to the control. The scuticocidal activity and respiratory burst activity were significantly enhanced in treated groups with or without parasite on weeks 2–4. However, the serum lysozyme activity was significantly enhanced from weeks 1 to 4. The protective response in terms of cumulative mortality was low in groups fed with enriched diet against parasite when compared to control. Therefore, we suggest that pomegranate enriched diet following challenge with P. dicentrarchi restores the altered hematological and biochemical parameters, and improves the innate immune system in olive flounder against P. dicentrarchi.

The effect of Suaeda maritima enriched diet on blood physiology, innate immune response, and disease resistance in olive flounder Paralichythys olivaceus against Miamiensis avidus on weeks 1, 2, and 4 was investigated. Feeding with any enriched diet and then challenging with M. avidus significantly increased white blood cells (WBC) on weeks 2 and 4; the red blood cells (RBC) significantly increased with 0.1% and 1.0% enriched diets on week 4. The hemoglobin (Hb) and hematocrit (Ht) levels significantly increased when fed with 0.1% and 1.0% supplementation diets on weeks 2 and 4. The mean corpuscular volume (MCV) did not significantly vary with any diet and time; however the mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) significantly increased with 0.1% and 1.0% supplementation diets on weeks 2 and 4. The leukocytes such as lymphocytes (Lym), monocytes (Mon), neutrophils (Neu) and biochemical parameters such as total protein (TP), glucose (GLU), and calcium (CAL) levels significantly increased in 0.1% and 1.0% supplementation diet fed groups on weeks 2 and 4. The serum lysozyme activity was significantly enhanced in 0.1% and 1.0% supplementation diet fed groups from weeks 1 to 4 when compared to the control (0% herbal extract enriched diet). The scuticocidal activity and respiratory burst activity were significantly enhanced when fish were fed with 0.1% and 1.0% supplementation diets from weeks 2 and 4. The protective effect in terms of cumulative mortality (50% and 40%) was low in groups on being fed with 0.1% and 1.0% supplemented diet. Therefore the present study suggested that 0.1% and 1.0% S. maritime-supplemented diets protect the hematological and biochemical parameters, improving the innate immunity, affording protection disease from M. avidus infection in olive flounder.

We investigated the effect of a mixed herb-enriched diet obtained from pomegranate Punica granatum, Dalmatian chrysanthemum Chrysanthemum cinerariaefolium, and mastic-leaved prickly-ash Zanthoxylum schinifolium on innate immune mechanisms (e.g., phagocytosis activity, respiratory burst activity, alternative complement activity, lysozyme activity, and disease resistance) of olive flounder Paralichthys olivaceus against the scuticociliate Philasterides dicentrarchi. All experimental groups were challenged with P. dicentrarchi (1 x 10(5) ciliates/mL) through intraperitoneal administration of the pathogen (50 microL) on day 1. On day 7, the infected groups were fed 0, 5, 50, and 100 mg/kg of the enriched diets. The innate immune parameters, cumulative mortality, and relative percent survival (RPS) were assessed at weeks 1, 2, 3, and 4. Administration of 50 or 100 mg/kg of the herbal-enriched diet enhanced immunity throughout the experimental period. However, at the 5-mg/kg dosage, the enriched diet did not enhance the innate immune estimates at any time. At doses of 50 and 100 mg/kg, administration of the diet preceding the challenge with P. dicentrarchi decreased the percentage cumulative mortality in the experimental groups and thereby increased RPS values. This study reports that administration of 50 or 100 mg/kg mixed herbal-enriched diet could positively influence the innate immune response to P. dicentrarchi and enhance the health status of olive flounder with respect to this microbe.

Lysozymes are the key molecules in innate immune system and possess high bactericidal properties. In the present study, a full-length c-type lysozyme cDNA has been cloned and characterized by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) techniques from kelp grouper, Epinephelus bruneus (Eb-CTL). The cDNA consists of 753 base pairs (bp) with a 432 bp open reading frame (ORF) that encodes 144 amino acids (aa) residues and polyadenylation signal sequence AATAAA. The deduced aa sequence polypeptide had a predicted molecular weight of 16 kDa and theoretical isoelectric point of 7.3. The deduced aa sequence have a long lysozyme domain which contains all catalytic sites and other conserved residues required for lysozyme activity. Pair-wise alignments showed a higher identity (76.4%) with Psetta maxima lysozyme and low identity (38.9%) with lysozymes of insect Bombyx mori. Interestingly, phylogenetic analysis revealed that the kelp grouper lysozyme was more closely related to other fish and vertebrate lysozymes. Quantitative real-time reverse transcriptase-PCR analysis showed that Eb-CTL transcripts are constitutively expressed in hematopoietic organs such as heart, spleen, and kidney after stimulation with LPS or infection with Vibrio anguillarum, indicating the involvement of Eb-CTL in the innate immunity of kelp grouper. This study is a first step on the genetics and functions of the c-type lysozyme in kelp grouper, and their role in anti-bacterial activity with reference to immunological properties which might have biotechnological applications.

This study determines the effect of diet enriched with the herb Baical skullcap Scutellaria baicalensis, and/or probiotics Lactobacillus sakei BK19 in rock bream, Oplegnathus fasciatus (32 ± 3 g) against Edwardsiella tarda. The changes in haematological parameters, innate immune response, and disease resistance were investigated after 1, 3, and 6 weeks. The white blood cell count (WBC: 10(4) mm(-3)), red blood cell count (RBC: 10(6) mm(-3)), and haemoglobin (Hb: g dl(-1)) levels significant increased (P < 0.05) with mixed diet on 3rd and 6th week and probiotics enriched diet on 6th week. The haematocrit (Ht: %) level significantly increased (P < 0.05) when fed with mixed diet on weeks 1-6. Interestingly, in mixed diet group the lymphocytes (LYM), monocytes (MON), and neutrophils (NEU) significantly increased from week 1-6. The eosinophils (EOS) significantly increased in all the treated groups. In the probiotics or mixed diet groups the total protein (TP: g dl(-1)) increased significantly on weeks 3 and 6. The serum lysozyme activity significantly was enhanced in all the treated groups indicating an increase in the innate immunity level. Serum complement, antiprotease activities, reactive oxygen species (ROS) and reactive nitrogen species (RNS) production significantly increased from week 1-6 with mixed diet. The maximum protection against E. tarda was recorded in mixed diet group with a minimum cumulative mortality of 20% and a high relative percent survival (RPS) of 72.84. In the probiotics and herbal diet groups the cumulative mortality was 25% and 35% and RPS was 68.63 and 59.42, respectively. This study indicates that administration of probiotics or mixed diets can effectively minimize the mortality and restore the altered hematological parameters and enhancing the innate immunity in O. fasciatus against E. tarda.

The effect of diets containing sodium alginate at 0, 0.5, 1.0, and 2.0 g kg⁻¹ following challenge with Streptococcus iniae in kelp grouper Epinephelus bruneus were assessed with reference to survival rate and innate immune parameters such as alternative complement, lysozyme, natural haemagglutination, respiratory burst, superoxide dismutase, and phagocytic activities on week 1, 2, and 4. Fish fed with sodium alginate containing diet at 1.0 and 2.0 g kg⁻¹ after being challenged with S. iniae had higher survival rates of 75% and 60%, respectively than those fed with control diet (0 g kg⁻¹). With any enriched diet the percentage of macrophages significantly decreased from week 1-4, while the percentage of neutrophils and lymphocytes significantly increased. The alternate complement activity, natural haemagglutination, and phagocytic activities of infected fish fed with sodium alginate containing diet at 1.0 g kg⁻¹ on week 2 and 1.0 and 2.0 g kg⁻¹ diets on week 4 were significantly higher when compared to the control. The lysozyme, respiratory bursts, and superoxide dismutase activities of fish fed with enriched diets at 1.0 and 2.0 g kg⁻¹ were significantly increased on week 2 and 4. We therefore recommend that at 1.0 or 2.0 g kg⁻¹ dietary administration of sodium alginate can enhance innate immunity and disease resistance in kelp grouper against S. iniae.

We report the effect of aqueous, ethanol, and methanol solvent leaf extracts of Punica granatum on innate immune mechanisms, such as phagocytosis activity, respiratory burst activity, alternative complement activity, lysozyme activity and functional immunity in terms of percentage cumulative mortality and Relative Percent Survival (RPS) in olive flounder Paralichthys olivaceus naturally infected with lymphocystis disease virus (LDV) after 8 weeks. Infected fish were intraperitoneally administered with 0, 5, 50, and 100 mg kg(-1) body weight of solvent extracts. In groups treated with 50 and 100 mg kg(-1) body weight, the chosen innate immune parameters significantly increased after 8 weeks when compared to 0 mg kg(-1) dose, but not with 5 mg kg(-1). Administration of P. granatum solvent extracts for 8 weeks significantly reduced the percentage mortality with the consequent increase in RPS. The results suggest that intraperitoneal administration of the leaf extracts of P. granatum at 50 or 100 mg kg(-1) dose clearly enhance the innate immune responses and disease resistance after 8 weeks in P. olivaceus against natural LDV infection.

The chemical constituents of decoction (individual) and concoction (mixed) of ethanolic leaf extracts from Azadirachta indica (neem) and Ocimum sanctum (tulsi) were analyzed by gas chromatography-­mass spectro-­ photometery (GC-­MS). Decoctions of A. indica and O. sanctum had 24 and 33 constituents, respectively. Mixed together, 26 compounds were identified. Four major (high percentage) compounds were identified in A. indica: n-­hexadecanoic acid (14.34%), phytol (19.96%), 9,12,15-­octa-­ decatrienoic acid, (Z,Z,Z)-­ (18.57%), and vitamin E (11.37%). Three major compounds were identified in O. sanctum: phenol,2-­methoxy-­3-­(2-­ propenyl) (15.32%), 9,12,15-­octadecatrienoic acid,(Z,Z,Z)-­ (16.94%), and 9,12,15-­octadecatrienoic acid, methyl ester,(Z,Z,Z)-­ (22.05%). Three major compounds were identified in the mixed extract: n-­hexadecanoic acid (16.58%), phenol,2-­methoxy-­3-­(2-­propenyl) (20.62%), and 9,12,15-­ octadecatrienoic acid,(Z,Z,Z) (25.98%). Four of the compounds in the mixed extract were new: eudesma-­4(14),11-­diene (0.18%), 6-­ azabicyclo[3.2.1]octane (0.51%), cyclohexane,1-­ethyl-­1-­methyl-­2,4-­ bis(1-­methylenyl)-­,[1S-­␣␣␣␣␣␣␣␣␣␣␣-­␣-­Elemen (0.77%), and globulol (1.45%). The mixed extract had a high level of antimicrobial activity against fish pathogens as indicated by zone of inhibition, minimum inhibitory concentration, and minimum bactericidal concentration.

The innate immunity and expression profiles of cathepsins D were determined in the goldfish (Carassius auratus) tissues after challenge with a fish pathogen Aeromonas hydrophila. The innate immunity of reactive oxygen species (ROS) and reactive nitrogen species (RNS) were determined by peripheral blood leucocytes. Blood and tissue samples of the muscle, gills, liver, kidney, heart, spleen, and intestine were sampled at 1, 3, 6 and 12 h post-infection for cathepsin D expression by semi-quantitative RT-PCR. The ROS and RNS production did not significantly increase at 1 h post-challenged goldfish. However, the ROS and RNS production was significantly increased after 3 h post-challenged fish compared to the control. The cathepsin D expression was found very low in muscle and kidney of the control fish, other tissues was not found the expression. A similar pattern was found in goldfish at 1 h post-challenge with A. hydrophila. However, at 3 h post-challenge goldfish, the cathepsin D expression was high only in the heart. At 6 h post-challenge goldfish, the cathepsin D expression was seen high all the tissues, except in the spleen. However, the expression was decreased at 12 h post-infection samples. This result was suggested that the goldfish infected with A. hydrophila decreased the innate immunity level in peripheral blood and expressed the cathepsin D in tissues.

We report the effect of aqueous-, ethanol- and methanol-solvent-derived extracts of three traditional Korean herbs, Punica granatum, Chrysanthemum cinerariaefolium and Zanthoxylum schinifolium, by monitoring the innate immune mechanisms, such as phagocytosis activity, respiratory burst activity, alternative complement activity and lysozyme activity and the functional immunity in terms of percentage mortality and relative percent survival (RPS) in olive flounder (Paralichthys olivaceus) against Uronema marinum (1 x 10(5)ciliates ml(-1)) for 30 days. Fish were intraperitoneally administered with 5, 50 and 100 mg kg(-1) body weight of each traditional Korean medicinal (TKM) solvent extract except the control and infected untreated groups. In all the treated groups at concentrations of 50 and 100 mg kg(-1) body weight, the chosen innate immune parameters were found significantly enhanced when compared to 0 mg kg(-1) dose. However, at 5 mg kg(-1) the tested immune parameters did not vary. Administration of TKM solvent extracts preceding the challenge with U. marinum for 30 days significantly reduced the percentage mortality with the consequent increase in RPS. Administration of 50 and 100 mg kg(-1) TKM solvent extracts clearly enhanced the innate immune responses and disease resistance in P. olivaceus against U. marinum.

The histophagous scuticociliate Philasterides dicentrarchi causes fatal scuticociliatosis in farmed olive flounder Paralichthys olivaceus. The average monthly prevalence of scuticociliatosis with P. dicentrarchi infections was very high from May to July (40+/-3 to 79+/-2%) in olive flounder at farms of Jeju Island, South Korea, from 2000 to 2004. The prevalence of mixed infection along with Vibrio spp. infection was higher (49+/-8%) than that of scuticociliatosis alone. To date no effective control measure for P. dicentrarchi infection has been described and large economic losses continue. In the present study 3 chemotheraputants (formalin, hydrogen peroxide and Jenoclean) were used. Among these, bath treatment with Jenoclean at a low concentration of 50ppm proved effective; the results were confirmed with in vitro motility assessments and morphological changes in P. dicentrarchi. A similar trend was noted following hydrogen peroxide treatment. However, formalin was only moderately effective at this concentration. Therefore, hydrogen peroxide and Jenoclean are the promising compounds effective at low concentrations with short application time.

This study reports the effect of aqueous, ethanol and methanol triherbal solvent extract from Azadirachta indica, Ocimum sanctum and Curcuma longa on innate immune mechanisms such as phagocytosis activity, respiratory burst activity, alternative complement activity and lysozyme activity and disease resistance in goldfish (Carassius auratus) against Aeromonas hydrophila. Fish were intraperitoneally injected with different doses of 0, 5, 50 and 100 mg kg(-1) body weight of each triherbal solvent extracts. The functional immunity in terms of percentage mortality and Relative Percent Survival (RPS) and innate immune response was assessed on week 1, 2 and 4 by challenging with live A. hydrophila (1 x 10(7) cells ml(-1)). All the chosen innate immune parameters were enhanced in the ethanol and methanol triherbal solvent extract treatment after week 2. However, the aqueous triherbal extract was enhanced only after week 4. The ethanol and methanol triherbal solvent extracts administration preceding the challenge with live A. hydrophila decreased the percentage mortality in the experimental groups with the consequence increase in RPS values. The study indicates that all the doses of ethanol or methanol triberbal solvent extracts could be positively influence the immune response and protect the heath status of goldfish against A. hydrophila infection.

Background: The genetic background of atopic dermatitis (AD) is not clearly understood. Interleukin (IL)-10 is a powerful Th-2 cell cytokine produced by lymphoid cells that exerts its function by inhibiting macrophage/monocyte and T-cell lymphocyte replication and secretion of inflammatory cytokines [IL-1, tumour necrosis factor-α (TNFA), IL-6, IL-8 and IL-12]. Objective: In an effort to discover additional polymorphism(s) in genes whose variant(s) have been implicated in total immunoglobulin E (IgE) level in AD patients, we scrutinized the single nucleotide polymorphisms (SNPs) in the IL10 gene as a potent candidate for contributing to the level of IgE in serum. Methods: We recruited 334 AD patients and assayed their serum total IgE levels using the LIPA-200 system. Four SNPs in the IL10 gene were genotyped using the single-base extension (SBE) method. Logistic regression analyses were performed with single polymorphisms and haplotypes (ht) to determine their association with the level of serum total IgE. Results: Genetic association analysis of total serum IgE in AD patients revealed that one of the IL10 ht, IL10-ht2, was associated with decreased serum total IgE in gene dose-dependent manner (P = 0.02–0.001). Conclusions: It was predicted that the inhibition of innate immunity by increased IL-10 production in IL10-ht2-bearing individuals might be associated with decreased total serum IgE levels among AD patients. The greater effects of IL10 ht on decreased total serum IgE levels suggest that the effect of IL-10 polymorphism might be the result of a combined genotype (ht) rather than single polymorphisms.