Oral administration of lactoferrin inhibits inflammation and nociception in rat adjuvant-induced arthritis

Lactoferrin (LF) is a ubiquitous protein which exists in milk, plasma, synovial fluids, cerebrospinal fluid and other biological fluids. LF is also well known as a natural immunomodulator. Recently, we found that bovine milk-derived LF (BLF) produced micro-opioid receptor-mediated analgesia. In this study, we examined whether oral administration of BLF causes anti-nociceptive and anti-inflammatory effects, and also whether it modulates LPS-induced TNF-alpha and IL-10 production in rat model of rheumatoid arthritis (RA), rat adjuvant arthritis. BLF was administrated once daily, starting 3 hr before (preventive experiment) or 19 days after (therapeutic experiment) adjuvant injection. In both experiments, BLF suppressed the development of arthritis and the hyperalgesia in the adjuvant-injected paw. The single-administered BLF produced a dose-dependent analgesia, which was reversed by naloxone, in the adjuvant arthritis rats. Both repeated and single administration of BLF suppressed TNF-alpha production and increased IL-10 production in the LPS-stimulated adjuvant arthritis rats. These results suggest that orally administered BLF has both preventive and therapeutic effects on the development of adjuvant-induced inflammation and pain. Moreover, the immunomodulatory properties of BLF, such as down-regulation of TNF-alpha and up-regulation of IL-10, could be beneficial in the treatment of RA. Thus, we concluded that LF can be safely used as a natural drug for RA patients suffering from joint pain.     

Effect of active immunization of pony mares against recombinant porcine inhibin alpha subunit on ovarian follicular development and plasma steroids and gonadotropins

Two pony mares were immunized against recombinant porcine inhibin alpha subunit three times with 39 day intervals. Clinical findings and endocrinological changes before immunization were taken as the control. The first significant rise in the anti-inhibin titre (P<0.05) in the circulation was found 27 days after the first injection. Maximum binding activity was reached by the 12th day after the second booster dose. The number of small, medium and large sized follicles had increased significantly compared to before immunization (11.75 +/- 4.30, 2.75 +/- 0.69 and 2.51 +/- 0.63 vs 6.50 +/- 1.43, 1.83 +/- 0.44 and 1.33 +/- 0.38, respectively), but the ovulation rate remained unchanged after immunization. The average plasma concentration of FSH and estradiol-17beta during the estrous cycle increased significantly (P<0.05) after immunization. These results suggest that immunization against inhibin is a useful tool to increase the number of ovarian follicles during the estrous cycle of pony mares. Moreover, the present study supported the concept that inhibin plays a major role in the control of follicular growth through its inhibitory effect on FSH secretion synergistically with steroid hormones.     

Isolation and expression of five-amino-acid-deleted variant of feline hepatocyte growth factor (HGF) cDNA

Hepatocyte growth factor (HGF) is a pleiotropic cytokine that stimulates a wide array of cellular targets, including hepatocytes and other epithelial cells, melanocytes, endothelial and hematopoietic cells. We have cloned a different form of cDNA, with a deletion of 15 base pairs predicted to result in the loss of 5 amino acids from the first kringle domain. To investigate the biological activity, original and deleted variant of feline HGF cDNAs were transiently expressed in COS-7 cells. Both recombinant feline HGFs showed almost the same dose-response curves in the stimulation of the growth of BNL CL.2 cells (a mouse hepatocyte cell line) and scatter activity of Madin-Darby canine kidney (MDCK) cells. The findings reported here suggest that the deleted variant of feline HGF has almost the same biological activity as the original in terms of the proliferation and scatter activity.     

Evaluation of four warming procedures to minimise heat loss induced by anaesthesia and surgery in dogs

OBJECTIVE: To evaluate the efficiency of four warming procedures, introduced after anaesthetic induction and continued during surgery, in minimising heat loss in anaesthetised dogs. DESIGN: Dogs were paired. One of each pair was a control; the other was subjected to one of four warming procedures. METHODS: Ninety-six dogs were involved in total. Pairs of dogs were matched for breed, hair length, and type of surgical procedure and placed adjacent to each other in a large temperature-controlled surgical theatre. One dog within each pair was assigned to one of four warming procedures that commenced immediately after anaesthetic induction. Group 1 (11 pairs) were placed on a purpose-designed prewarmed (41 degrees C) electrically heated pad. Group 2 (18 pairs) were placed on a prewarmed electric heat pad (41 degrees C), cocooned by four wrapped water bottles (initially 41 degrees C) and subjected to radiant heat (150 watt lamp placed 50 cm away from the head of the dog). Group 3 (11 pairs) were surrounded by a forced air warming mattress (set at 43 degrees C). Group 4 (8 pairs) were connected via the anaesthetic breathing circuit to a heater/humidifier (set at 41 degrees C). Rectal temperature measurements were recorded every 15 min for the first 3 h of anaesthesia. The fall in rectal temperature of the control dog was subtracted from the fall in temperature of the treatment dog and this measurement was used to assess the efficacy of the various warming procedures. RESULTS: The mean rectal temperature of unheated 'control' dogs decreased 1.9 +/- 0.6, 1.4 +/- 0.4 and 1.1 +/- 0.4 degrees C over the first, second and third hour respectively. After 3 h the temperature fall differential for all groups were 0.7 +/- 0.7 (Group 1), 3.1 +/- 1.1 (Group 2), 2.4 +/- 1.1 (Group 3) and 1.0 +/- 1.1 degrees C (Group 4). Thus the group 2 procedure was the most successful in preventing a drop of temperature followed by groups 3, 4 and 1. CONCLUSION: Large dogs undergo significant reduction in core body temperature especially during the first 2 h of anaesthesia and surgery. Supplementary warming substantially reduces this fall in body temperature, although certain warming procedures were found to be more effective than others.     

Mice cloned by nuclear transfer from somatic and ntES cells derived from the same individuals

The current success rate of cloned mice from adult somatic cell nuclei is very low, whereas it is relatively high for cloned mice from ES cell nuclei. In this experiment, we examined whether the success rate of cloning from somatic cells could be improved via nuclear transfer embryonic stem cells (ntES cells) established from somatic cell nuclei. We obtained 11 cloned mice and 68 ntES cell lines from the somatic cell nuclei of 7 mice, and cloned 41 mice were cloned from the ntES cell nuclei. Unexpectedly, the overall success rate of cloning from ntES cell nuclei in this series was no better than when using somatic cell nuclei. Interestingly, full-term cloned mice were produced only via ntES cells from two individuals, but not by direct nuclear transfer from the somatic cells, and vice versa. Ultimately, we were able to obtain clone mice from 6 out of 7 individuals using either somatic cells or ntES cells. Thus, although ntES cells as donor nuclei do not absolutely assure a better success rate for mouse cloning than somatic cells, to preserve and clone valuable individuals, we recommend that ntES cell lines be established. These can then be used as an unlimited source of donor nuclei for nuclear transfer, and thus complement conventional somatic cell nuclear transfer cloning approaches.     

Evaluation of hemodynamic measurements, including lithium dilution cardiac output, in anesthetized dogs undergoing ovariohysterectomy

OBJECTIVE: To measure cardiac output in healthy female anesthetized dogs by use of lithium dilution cardiac output and determine whether changes in mean arterial pressure were caused by changes in cardiac output or systemic vascular resistance. DESIGN: Prospective clinical study. ANIMALS: 20 healthy female dogs. PROCEDURE: Dogs were anesthetized for ovariohysterectomy. Ten dogs breathed spontaneously throughout anesthesia, and 10 dogs received intermittent positive-pressure ventilation. Cardiovascular and respiratory measurements, including lithium dilution cardiac output, were performed during anesthesia and surgery. RESULTS: Mean arterial pressure and systemic vascular resistance index were low after induction of anesthesia and just prior to surgery and increased significantly after surgery began. Cardiac index (cardiac output indexed to body surface area) did not change significantly throughout anesthesia and surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Results provide baseline data for cardiac output and cardiac index measurements during clinical anesthesia and surgery in dogs. Changes in mean arterial pressure do not necessarily reflect corresponding changes in cardiac index.     

Effects of different soybean proteins on lipid digestion and growth of yellowtail <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis>

Two experiments were conducted to determine whether the undigested high molecular fraction (HMF) of soybean protein reduces growth of yellowtail through impairment of lipid digestion and absorption. Soybean meal (SBM), soy protein isolate (SPI), digested SPI (DSPI), and digested and purified SPI (DPSPI) were used as experimental diets. In experiment 1 (exp. 1), yellowtail were fed 4 diets with aforementioned soybean products for 3 weeks. In experiment 2 (exp. 2), the fish were fed soybean protein or fish meal (FM) diets for 8 weeks. Growth performance of DPSPI-fed fish was better than that of fish fed other soybean proteins, and comparable to that of fish fed with FM. Plasma and tissue lipid levels of DPSPI-fed fish tended to be higher than those of fish fed other soybean proteins. Significantly lower intestinal lipid levels and higher bile acid levels were found in DPSPI-fed fish than in fish fed with SBM, SPI, and DSPI. These findings indicate that one of the factors contributing to growth depression in yellowtail fed with SBM-based diets is the negative effect of undigested HMF of soybean protein on bile acid levels and lipid digestion.     

Association between bacterial community structures and mortality of fish larvae in intensive rearing systems

Bacterial community structures were analyzed in water used for rearing fish larvae by fluorescence in situ hybridization. In Experiment 1, red sea bream Pagrus major larvae were reared in two commercial seed production tanks. The survival rate in Tank 1 was higher than in Tank 2, even though phytoplankton, Nannochloropsis sp., was added to both tanks. In Tank 2, γ-proteobacteria became dominant (∼70% of total bacteria) on day 13, there after heavy larval mortalities occurred. In Tank 1, however, α-proteobacteria and the Cytophaga-Flavobacterium cluster were predominant from day − 1 until day 13; no significant mortality was recorded. In Experiment 2, marble goby Oxyeleotris marmoratus larvae were cultured with or without Nannochloropsis sp. At the end of the experiment, larval survival rates in aquaria with Nannochloropsis sp. were significantly (P <0.05) higher than those without. In rearing water without Nannochloropsis sp., γ-proteobacteria increased during rearing. In rearing water with Nannochloropsis sp., α-prote obacteria and the Cytophaga-Flavobacterium cluster were predominant at the beginning of the experiments and the relative abundance of γ-proteobacteria was maintained at a lower level throughout the experiments. The predominance of α-proteobacteria and the Cytophaga-Flavobacterium cluster appears to be a good indicator of successful larval production.     

Purification and characterization of lipoxygenase from Pleurotus ostreatus

Lipoxygenase was purified homogeneously from cups of Pleurotus ostreatus by Sephacryl S-400 HR gel filtration, Dyematrex Green A affinity, and DEAE-Toyopearl 650M ion-exchange chromatographies. The molecular weight of the enzyme was estimated to be 67,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 66,000 by gel filtration; the isoelectric point was pH 5.1. The optimum pH and temperature of the enzymatic activity were 8.0 and 25 degrees C, respectively. The enzyme contained non-heme iron, and a thiol group seemed to be involved in its activity. The K(m), V(max), and k(cat) values of the enzyme for linoleic acid were 0.13 mM, 23.4 micromol.min(-1).mg(-1), and 25.7 s(-1), respectively. The enzyme showed high specificity toward linoleic acid. When linoleic acid was incubated with the enzyme, 13-hydroperoxy-9Z,11E-octadecadienoic acid was found to be the main oxidative product.