Maturation and spawning induction in Hawaiian opihi Cellana spp. by hormone GnRH.

Gonadotropin releasing hormone (GnRH) is a neuropeptide hormone whic plays an important role in control of the reproduction among vertebrates and some work is showing that it is active in invertebrate species. We matured and spawned the Hawaiian opihi Cellana spp. by using salmon GnRH analogue. In the first experiment on maturation, twelve adult opihi (3.18+/-0.23cm, shell length) were injected weekly with salmon GnRHa at dose of 250ng.g-1 body weight (BW) for a 6-week period. Gonad development was assessed using gonadosomatic index (GSI) and histological techniques. GSI incorporated with histological analysis showed that the gonads reached full maturation after 4 weeks. GSI increased significantly (31%) in comparison to the control group of saline solution injection did not produce any maturity in opihi. For the spawning trial, eight ripe opihi were administered a single injection at dose of 1000ng.g-1 BW. The results showed that an average of 33% of the animal spawned in 4-6h after hormone injection. Fertilized eggs developed through embryo and to the com plete larval stage of veligers. Our preliminary findings in this study provide a new aspect of hormone controlling reproduction of opihi, which could be applied in opihi aquaculture.

Comparison of techniques to control the aggressive environmental invasive species Galenia pubescens in a degraded grassland reserve, Victoria, Australia.

Across many southern regions of Australia, native grasslands have become seriously threatened by human activity, with only a fraction of the original areas remaining undisturbed. In particular, the introduction and establishment of exotic invasive weeds has caused significant degradation to the ecosystems in these areas by contributing to a decrease in native plant density and diversity, and this has ultimately led to major changes to the ecosystem structure and function. One such example is Galenia pubescens. Our objective of this study was to assess the effectiveness of four different attempts to control G. pubescens: herbicide control with glyphosate; organic herbicide control with pine oil; the application of mulch; and the addition of seeds of native species to the seedbank. Results shows that any one single control strategy is insufficient to control G. pubescens, and, in addition, it has shown that regeneration of native vegetation is limited unless direct seeding is applied. There was a strong indication that a combined strategy employing more than two of the aforementioned techniques is likely to be the most effective approach, at least in the short term. Underscoring the complexity of this task, our analysis on foliage cover of G. pubescens shows that the interaction of pine oil and glyphosate treatments appeared to be very effective after six months, but were not so effective after 18 months. By contrast, seeding with native seeds was not particularly effective at six months, but its longer-term contribution appears to be effective at 18 months. Further, our results obtained from the seedbank abundance study indicate that time alone was not a significant factor in restoration of the grasslands (p = 0.165); however there were interactions with time, shown by time*glyphosate (p = 0.008) and time*seeding (p = 0.016). Both interactions indicated that the applications of glyphosate and seeding were more beneficial after 18 months compared to six months. However, full regeneration of invaded native grasslands may not be possible unless further restoration programs are re-implemented after the first cycle of G. pubescens' treatments have been completed.

A 50000-dalton anorexigenic protein from urine.

A potent, homogeneous, 50 kDa protein which transiently depresses food intake of rats up to 24 h after injection was isolated from rat urine. It was prepared from an ethanolic benzoic acid precipitate of urine. The material was subjected to DEAE-Sephacel chromatography, whereupon four fractions (DEAE pools a, b, c, and d) were obtained. The latter three contained anorexigenic activity. DEAE pools b and c were subjected to further purification on columns of Sephacryl S-300 eluted with buffer containing 8 M urea. One of the subfractions of DEAE pool b contained only one protein as judged by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. This protein had a molecular weight of 50,000 and was active in reducing 24 h food intake at a dose of 130 micrograms/100 g bodyweight.

Improved isolation of a 50 kDa anorexigenic protein from rat urine.

In prior work, a 50 kDa protein was purified to homogeneity from rat urine. This protein reduces food intake when injected into rats and is the only natural substance other than satietin known to be effective for long (24 hour) time periods and which does not make animals ill. However, when attempts were made to repeat the purification, contamination appeared in the 50 kDa fraction. The present contribution documents successful reisolation of the 50 kDa anorexigen by an improved method. Reisolation involved Cibacron blue-Sepharose, DEAE-Sephacel and Sephacryl S-200 chromatography, and SDS disc preparatory electrophoresis. The reisolated 50 kDa anorexigen contains no detectable carbohydrate. Partially purified preparations of the 50 kDa anorexigen were fragmented with trypsin and proteinase K without loss of anorexigenic activity. It is concluded that the 50 kDa anorexigen may be reproducibly purified to homogeneity and may contain within its amino acid sequence a peptide which is the basis of its anorexigenic activity.

Healthful new oil from macadamia nuts.

Fatty acid profiles were obtained for macadamia nut oil and several other cooking oils. Macadamia nut oil contained the highest level (79%) of monounsaturated fatty acids. Macadamia nut oil is low (4%) in the omega-6 fatty acid 18:2n-6 and saturated fatty acids. Fatty acid profiles for popcorn cooked in several oils were obtained for comparison because of public awareness and concern over high levels of saturated fat found in popcorn sold in theaters. The nutritional implications of using macadamia nut oil are discussed.

Bullous eruption of systemic lupus erythematosus.

A bullous eruption is now recognized as a specific complication of systemic lupus erythematosus (SLE). We described a 22-year-old woman with SLE whose vesiculobullous eruptions exacerbated with a flare of visceral manifestations of the disease and resolved when the systemic disease was controlled by oral prednisolone.

[Clinical evaluations of flomoxef in respiratory tract infections].

Flomoxef (FMOX, 6315-S), a new antibacterial drug, was administered to 9 cases with respiratory tract infections for a duration of 8 approximately 16 days at a daily dose of 2 g. Diagnosis of these patients were bronchopneumonia 5 cases, chronic bronchitis 3 cases and acute bronchitis 1 case. From transtracheal aspiration several organisms were isolated; Haemophilus influenzae was isolated in 3 cases, Streptococcus pneumoniae in 3 cases, H. influenzae plus Branhamella catarrhalis in 1 case, Streptococcus dysgalactiae plus Neisseria meningitidis in 1 case and Corynebacterium pseudodiphtheriticum in 1 case. The clinical efficacy was good in all 9 cases, the efficacy rate was 100%. All the bacteria were eliminated. Side effects were not observed. From these results, it appears that FMOX is a valuable drug in the treatment of respiratory tract infections.

[A case of severe juvenile pulmonary tuberculosis associated with malnutrition in special reference to nutritional assessment].

We reported a case who had suffered from severe pulmonary tuberculosis in association with severe malnutrition. A 19-year-old man was admitted complaining of high fever, productive cough and body weight loss. Before admission, he worked as a cook for long hours a day and had meals irregularly. On admission, he was cachexic. Coarse crackles were auscultated on the both lung field, and the liver was enlarged. Chest X-ray revealed multiple ringed and mottled shadows mainly in bilateral upper and middle lung field. Laboratory examination revealed inflammatory reaction defined by an increase of white blood cells, accelerated erythrocyte sedimentation rate and positive CRP, impairment of liver function and cell-mediated immunity, hypoxemia and restrictive lung dysfunction. Ziehl-Neelsen stain of sputum showed a large number of acid fast bacilli. The assessment of his nutritional status showed decrease in anthropometric measurements, visceral proteins and the Fischer ratio, which suggested that he was in the status of protein-calorie malnutrition in association with amino acid imbalance. He was treated with nutritional therapy in addition to antituberculous drugs, which improved his clinical symptoms, laboratory data and nutritional assessment. These results suggested the importance of nutritional assessment and nutritional therapy based on it.

[An autopsy case of atypical mycobacteriosis accompanied with chest wall deformity].

An autopsy case of 63-year old male of atypical mycobacteriosis with severe scoliosis was reported. He was diagnosed as atypical mycobacteriosis in 1981 and controlled by antimycobacterial treatment. But since 1989, dyspnea and sputum have been increasing, new abnormal shadow in company with progression of curvature of the scoliosis was revealed on the chest roentgenogram. In spite of various treatments for mycobacteriosis, he died of respiratory failure. The aggravation of this mycobacteriosis brought on advancement of malnutrition and impairment of cell mediated immunity. Findings of autopsy showed the remarkable chest wall deformity and tuberculous change of the lung parenchyma. In this case, chest deformity was thought as the major risk factor of onset of mycobacterial mycobacteriosis.

[Three cases of young adults advanced pulmonary tuberculosis due to patient's and doctor's delay].

Three young adults advanced pulmonary tuberculosis due to delay of therapy were reported. In case 1 and 3, their delays resulted from difficulty in diagnosis, in case 2, from neglecting medical counseling. Each cases revealed bilateral diffuse shadows on chest roentgenograms on admission, which were typical shadows of advanced pulmonary tuberculosis. Malnutrition might contribute to the development of the diseases, which were improved by anti-tuberculosis therapy and hyperalimentation therapy. These cases were suggested some clinical problems characteristic of pulmonary tuberculosis of young adults.

Omega-3 fatty acids in Hawaii seafood.

Omega-3 fatty acids, the most potent of which are found in seafood, are of interest because of their effects on cardiovascular and inflammatory diseases and their possible effects on cancer. However, consumers in Hawaii wishing to increase their dietary omega-3 to omega-6 fatty acid ratio are faced with the difficulty that several types of seafood popular in Hawaii and aquacultured seafood new in the marketplace have unknown omega-3 fatty acid levels. The purpose of this work is to determine omega-3 fatty acid levels of selected seafood and fish oil capsules. Several seafoods and some over-the-counter fish oil capsules were sampled and analyzed. Aku eggs, aquacultured hamachi (yellowtail jack from Japan), one sample of turbot, and EPA Plus, Promega, and Omega-3 Super EPA capsules were found to contain high levels of omega-3 fatty acids. Levels were comparable to those in the fatty fishes such as salmon and mackerel. Butterfish, mahimahi eggs; other fish oil capsules (ProEPA and Omega 3) contained moderate levels of omega-3 fatty acids. Fish cakes, seaweed, several lean fishes, and cod liver oil capsules had small quantities of omega-3 fatty acids. It appeared that the omega-3 fatty acid content of aquacultured species studied was significantly higher than in wild caught species. There was a substantial difference between claimed and actual omega-3 fatty acid levels in commercially available fish oil capsules. These findings can help consumers when selecting types of seafood for their diet that are good sources of omega-3 fatty acids.

[Expression of activation antigen, CD69, on human local eosinophils].

The activation antigen, CD69, has been shown to be expressed on activated lymphocytes. CD69 has also been shown to be associated with the signal transduction process. Recently, we demonstrated the expression of CD69 on lung eosinophils obtained from the bronchoalveolar lavage fluid of patients with eosinophilic pneumonia. In this study, we performed flowcytometric analysis of CD69 expression on eosinophils obtained from the bronchoalveolar lavage fluid, pleural effusion or peripheral blood of various patients. Whatever the disease, local eosinophils expressed significant levels of CD69, whereas most peripheral blood eosinophils did not express CD69. These findings suggest that CD69 is a marker for local eosinophils and that local eosinophils may differ functionally from peripheral blood eosinophils.

[Expression of activation antigen CD69 on eosinophils].

The antigen, CD69, has been shown to be expressed on activated lymphocytes. CD69 has also been shown to be associated with signal transduction process. Recently, we demonstrated the expression of CD69 on eosinophils at inflammatory sites. Whatever the disease, lung eosinophils obtained from the bronchoalveolar lavage fluid or pleural effusion expressed significant levels of CD69, whereas most peripheral blood (PB) eosinophils did not express CD69. Further, in vitro stimulation of PB eosinophils by IL-3, IL-5, GM-CSF and PMA induced CD69 expression on PB eosinophils. These findings suggest that CD69 expression on eosinophils is restricted to local inflammatory sites and CD69 may be a useful marker for activated eosinophils at inflammatory sites.

Isolation of a fibrinolysis enzyme activator from commercial bromelain.

A fibrinolysis enzyme activator was isolated from commercial bromelain. This enzyme preparation is used medicinally. A by-product of bromelain production, the acetone still residue, was used as a starting material to develop an isolation scheme which began with reversed phase chromatography, was followed by ion exchange chromatography, and ended with ultrafiltration. It had polarity and ion exchange properties similar to those of a neutral amino acid. The active substance has no intrinsic fibrinolytic activity but appears to enhance the activity of one of the fibrinolytic enzymes. The presence of the fibrinolysis enzyme activator in commercial bromelain may explain some of the physiological and clinical effects observed following the oral administration of the enzyme preparation.

gamma-Carboxyglutamic acid, a component in human pediatric bladder stones containing calcium salts.

The amino acid gamma-carboxyglutamic acid (Gla) has been previously detected in the vitamin K-requiring blood clotting factors, proteins of calcified vertebrate tissue, renal tissue, plasma protein C, ectopic calcifications, and calcium-containing renal calculi. This paper reports the presence of Gla in the EDTA-soluble, non-dialyzable material recovered from human pediatric bladder stones containing calcium salts. In bladder stones composed of calcium oxalate, uric acid and ammonium acid urate, 73 Gla residues per 1,000 amino acid residues were detected. Bladder stones composed of calcium oxalate, uric acid, ammonium acid urate, and hydroxyapatite contained 48 Gla residues per 1,000 amino acid residues present. No Gla was detected in the predominantly magnesium ammonium phosphate (struvite) bladder stones. These results with human bladder stones from children under 10 years of age are consistent with the observations from adult patients in which Gla was detected in the calcium-containing renal calculi but not in the non-calcium-containing renal calculi. The present study adds to the growing body of information concerning the possible role of Gla in normal and abnormal calcium metabolism.

Isolation of a protease from sea urchin eggs before and after fertilization.

Upon fertilization, sea urchin eggs (Stronglyocentrotus pupuratus) release a protease into the surrounding sea water. This protease is in a particulate form which can be solubilized. The soluble form was purified by affinity chromatography on columns of immobilized soybean trypsin inhibitor. The purified enzyme is similar to bovine trypsin both in molecular weight (22500) and in susceptibility to inhibitors such as diisopropyl phosphofluoridate and soybean trypsin inhibitor. In contrast, extracts of unfertilized eggs appear to contain an inactive form of the enzyme which can be activated by dialysis at pH 4.6. The enzyme, as purified from extracts activated in this manner, was similar in its properties to that from fertilized eggs.

Nutrient, fatty acid, amino acid and mineral analysis of natural prey of the Hawaiian monk seal, Monachus schauinslandi.

Proximate nutrients, gross energy content, mineral, amino acid and fatty acid composition were determined for teleost, cephalopod and crustacean prey of the Hawaiian monk seal. Crude protein was highest in the octopus, Octopus cyanea (80.0%), crude fat was highest in the Muraenid teleost, Gymnothorax eurostus (14.1%), whereas crude ash was highest in the lobster, Panulirus marginatus (11.6%). Gross energies ranged from 4.0 +/- 0.01 kcal g-1 in the Labrid teleost Bodianus bilulunatus to 6.0 +/- 0.12 kcal g-1 in the moray eel, Gymnothorax undulatus. Essential amino acids occurred in lower concentrations as a percentage of the total amino acids (35.8 +/- 2.6%) than non-essential amino acids (64.2 +/- 2.6%), but the ratio of individual amino acids to total amino acid concentrations were similar to those required by some monogastric terrestrial species and fingerling salmon. The fatty acid concentrations varied widely among species (range = 1.2-16.5 mg 100 mg-1); however, the teleosts had higher total fatty acids than the non-teleosts. This study indicates that, from a nutritional standpoint, some prey may be more beneficial to the Hawaiian monk seal; however, these prey are not necessarily the most abundant or available to some populations of the monk seal.

[Effect of short-term hypoxia on blood platelet function tests of normal subjects].

We studied how smoking and hypoxia affected blood platelet functions. Eighteen normal subjects (ten nonsmokers and eight smokers) inspired 13% oxygen gas and underwent platelet function tests (blood platelet count, MPV, maximum platelet aggregation rate, plasma beta-TG, plasma PF-4) measured just before inspiration and 30 minutes and 60 minutes after inspiration of the gas. Four principal results were obtained. First, breathing room air, blood platelet count and maximum platelet aggregation rate of smokers were significantly increased compared to those of non-smokers. Second, platelet counts and MPV were not affected by hypoxia in either group. Third, the maximum platelet aggregation rate of smokers was significantly decreased by hypoxia. Fourth, in both groups, the plasma levels of beta TG and PF-4 were decreased 30 minutes after inspiration of the 13% oxygen gas. In non-smokers, 60 minutes after inspiration, these plasma levels recovered to the primary levels. Our study showed that blood platelet functions of normal subjects were affected by hypoxia, and the effects of hypoxia on these functions in these two groups were markedly different.