Isolation of two anti-inflammatory and one pro-inflammatory polyunsaturated fatty acids from the brown seaweed Undaria pinnatifida

Two anti-inflammatory omega-3 polyunsaturated fatty acids (PUFAs) of stearidonic acid (SA) and eicosapentaenoic acid (EPA) and one pro-inflammatory omega-6 PUFA of arachidonic acid (AA) were isolated from the edible brown seaweed Undaria pinnatifida. SA was active against mouse ear inflammation induced by phorbol myristate acetate, with IC50 values of 160, 314, and 235 microg per ear for edema, erythema, and blood flow, respectively. EPA was also active against edema, erythema, and blood flow, with IC50 values of 230, 462, and 236 microg per ear, respectively. Although AA at low concentrations showed anti-inflammatory activities when measured 10 h later, AA doses of more than 243 microg per ear induced inflammatory symptoms 1 h later. Mature thalli generally had larger amounts of PUFAs than young thalli. The algal blade contained more omega-3 PUFAs than were found in other parts, while the holdfast contained extremely high amounts of AA. Late-season thalli showed increased amounts of PUFAs, especially AA.     

MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.     

Migration of anadromous white-spotted charr <Emphasis Type="Italic">Salvelinus leucomaenis</Emphasis>, as determined by otolith strontium: calcium ratios

ABSTRACT:   The migratory history of anadromous white-spotted charr Salvelinus leucomaenis collected from Japanese coastal waters, was examined in terms of strontium (Sr) and calcium (Ca) uptake in the otolith, by means of wavelength dispersive X-ray spectrometry using an electron microprobe. Otolith Sr concentration or Sr : Ca ratios of anadromous S. leucomaenis , fluctuated strongly along the life history transect in accordance with the migration (habitat) pattern from sea to fresh water. The anadrmous S. leucomaenis showed phase L (low Sr : Ca ratio) from the core to the point 1000–2500 µm distant, averaging 1.3 × 10⁻³ to 2.7 × 10⁻³ and thereafter, the ratios increased sharply, being higher than 5.0 × 10⁻³ to 10.0 × 10⁻³. These findings indicated that otolith Sr : Ca ratios reflected individual life histories, enabling a sea habitat to be identified from a freshwater habitat in this species.     

Toxicity of the antifouling biocide Sea-Nine 211 to marine algae, crustacea, and a polychaete

We evaluated the acute toxicity of the antifouling biocide Sea-Nine 211 to the algae Chaetoceros calcitrans, Dunaliella tertiolecta, Tetraselmis tetrathele, and Skeletonema costatum, the crustacea Tigriopus japonicus and Portunus trituberculatus, and the polychaete Perinereis nuntia. The algae, and especially the diatoms C. calcitrans and S. costatum, were sensitive to Sea-Nine 211 toxicity, with the average acute toxicity values being 0.32, 3.9, 1.6, 0.22, 1.6, 12, and 27 μg/l for C. calcitrans, D. tertiolecta, T. tetrathele, S. costatum, T. japonicus, P. trituberculatus, and P. nuntia, respectively. A sediment toxicity test for Sea-Nine 211 using the polychaete P. nuntia revealed demonstrated that the 14-day median lethal concentration was 110 μg/kg dry-wt sediment and that growth was the most sensitive indicator. The chronic toxicity values of Sea-Nine 211 for the diatoms C. calcitrans and S. costatum were within the range of reported Sea-Nine 211 concentrations in seawater in coastal Japan, and the toxicity values for P. nuntia were within the reported concentrations in sediment. Based on these results, Sea-Nine 211 may have toxic effects on some sensitive species residing in the coastal areas of Japan, but the ecological risk posed by Sea-Nine 211 would appear to be confined to a limited area of Japanese coastal waters.     

Experimental dual infection of pigs with an H1N1 swine influenza virus (A/Sw/Hok/2/81) and Mycoplasma hyopneumoniae

Dual infection of pigs with swine influenza virus (SIV) and Mycoplasma hyopneumoniae was carried out to compare the clinical and pathological effects of dual infection in caesarian derived and colostrums deprived (CDCD) pigs, with that of a single infection with M. hyopneumoniae. In Experiment 1, 40-day-old CDCD pigs were inoculated only with SIV (A/Sw/Hok/2/81, H1N1). The virus was isolated from nasal swabs for 5-6 days. None of these pigs showed clinical signs of infection throughout the experimental period. These results suggested that this strain can infect pigs but is only slightly pathogenic when it is inoculated singly to a CDCD pig. In Experiment 2, 60-day-old CDCD pigs were inoculated with M. hyopneumoniae and then were inoculated with SIV (A/Sw/Hok/2/81) at 1 week (MHYO-7d-SIV-7d group) or 3 weeks (MHYO-21d-SIV-7d group) after M. hyopneumoniae inoculation. Macroscopically, dark red-to-purple lung lesions were observed in all of pigs at 14 or 28 days post-inoculation. Percentages of dark red-to-purple lung lesions in dual infection groups (MHYO-7d-SIV-7d group: 18.7 +/- 4.2%, MHYO-21d-SIV-7d group: 23.0 +/- 8.0%) were significantly (P < 0.05) increased compared to those of each control group in which pigs were inoculated only with M. hyopneumoniae (MHYO-14d group: 4.7 +/- 2.9%, MHYO-28 group: 3.3 +/- 2.4%). Microscopically, bronchial epithelial lesions (epithelial disruption, degeneration, hyperplasia and formation of microabscess) were frequently observed in dark red-to-purple lung lesions of only the dual infection groups. These results demonstrate that the lung lesion of pigs inoculated with M. hyopneumoniae and SIV is more severe than that of pigs inoculated only with M. hyopneumoniae.     

Intraocular vaccination with an inactivated highly pathogenic avian influenza virus induces protective antibody responses in chickens

Because it is expected to induce cross-reactive serum and mucosal antibody responses, mucosal vaccination against highly pathogenic avian influenza (HPAI) is potentially superior to conventional parenteral vaccination. Here, we tested whether intraocular vaccination with an inactivated AI virus induced protective antibody responses in chickens. Chickens were inoculated intraocularly twice with 10⁴ hemagglutination units of an inactivated H5N1 HPAI virus. Four weeks after the second vaccination, the chickens were challenged with a lethal dose of the homologous H5N1 HPAI virus. Results showed that most of the vaccinated chickens mounted positive antibody responses. The median serum hemagglutination inhibition titer was 1:80. Addition of CpG oligodeoxynucleotide 2006 or cholera toxin to the vaccine did not enhance serum antibody titers. Cross-reactive anti-hemagglutinin IgG, but not IgA, was detected in oropharyngeal secretions. In accordance with these antibody results, most vaccinated chickens survived a lethal challenge with the H5N1 HPAI virus and did not shed the challenge virus in respiratory or digestive tract secretions. Our results show that intraocular vaccination with an inactivated AI virus induces not only systemic but also mucosal antibody responses and confers protection against HPAI in chickens.     

Assessment of the accuracy of testicular dysfunction detection in male donkey (Equus asinus) with the aid of colour-spectral Doppler in relation to plasma testosterone and serum nitric oxide levels

This study aimed to determine the usefulness of colour and pulsed Doppler modes for the accurate diagnosis of donkeys suffering from subfertility to determine whether testicular vascularity assessment could be an indicator for sperm functionality. The study sample was composed of 10 male donkeys with normospermia (control group) and 10 donkeys with hypospermia. Animals underwent scrotal circumference measurement, testicular Doppler examination, seminal evaluation, blood sampling and hormonal assay. Semen volume and concentration were significantly (p ≤ .05) lower in the subfertile group (30.25 ± 1.22 ml and 89.44 ± 2.55 × 10⁶/ml) as compared with the control group (82.76 ± 1.65 ml and 452.78 ± 1.25 × 10⁶/ml), and total sperm/ejaculation was significantly (p ≤ .05) higher in the normal donkeys (28.30 ± 2.32 × 10⁹/total ejaculated) as compared with the subfertile group. Intratesticular coloured area showed a marked decline in the hypospermic males. There was no significant difference between the two groups in testosterone level, although the normal group showed an increase in nitric oxide metabolites. Both Doppler indices of the three branches of the testicular artery were elevated significantly (p ≤ .05) in abnormal donkeys, whereas Doppler peak systolic and end-diastolic velocities were increased in the normal group. Male donkeys with subfertility demonstrated lower arterial vascularity parameters in the form of intratesticular coloured area and blood flow rate; therefore, the most optimal parameters for differentiating subfertile hypospermic from normospermic donkeys were found to be the two Doppler indices, velocities parameters, testicular blood flow rate and nitric oxide levels.     

A canine case of skull base meningioma treated with hydroxyurea

An 11-year-old female miniature schnauzer was tentatively diagnosed with the skull base meningioma, based on several examinations. Because surgical treatment was difficult, and outpatient radiation therapy was not available in the local area, chemotherapy with hydroxyurea combined with dexamethasone was selected. The patient's clinical symptoms improved after one week of treatment, and the tumor size was obviously reduced on MRI performed 37 days after treatment began. The patient received hydroxyurea for 7 months, with symptoms remaining stable, and the tumor re-increased to almost the same size at 7 months as that at the initial examination. At that time, hydroxyurea was discontinued. The patient died from pulmonary edema 14 months after treatment began. Pathologically, the tumor was diagnosed as a meningioma.     

Sequence of the domestic duck (Anas platyrhynchos) growth hormone-encoding gene and genetic variation in the promoter region

The duck growth hormone encoding gene and its promoter region were amplified by polymerase chain reaction (PCR). A total of 5.25 kb were cloned and sequenced. Duck growth hormone (GH) consists of five exons and four introns and is structurally similar to mammalian and chicken GH gene. Although the distal region of duck GH promoter showed no similarity to chicken and turkey promoters, the proximal region of the promoter contained two putative Pit‐1 binding sequences, and showed similarity to chicken and turkey GH promoters. Genetic variation was detected at five positions of the promoter region. The results of this study indicate that the expression of duck GH is likely regulated in a similar manner to that of chicken GH via enhancer‐type cis‐acting elements and the presence of genetic variation in the duck GH gene may be applicable to marker‐assisted selection.