Enrofloxacin enhances the effects of chemotherapy in canine osteosarcoma cells with mutant and wild-type p53.

Adjuvant chemotherapy improves survival time in dogs receiving adequate local control for appendicular osteosarcoma, but most dogs ultimately succumb to metastatic disease. The fluoroquinolone antibiotic enrofloxacin has been shown to inhibit survival and proliferation of canine osteosarcoma cells in vitro. Others have reported that fluoroquinolones may modulate cellular responses to DNA damaging agents and that these effects may be differentially mediated by p53 activity. We therefore determined p53 status and activity in three canine osteosarcoma cell lines and examined the effects of enrofloxacin when used alone or in combination with doxorubicin or carboplatin chemotherapy. Moresco and Abrams canine osteosarcoma cell lines contained mutations in p53, while no mutations were identified in the D17 cells or in a normal canine osteoblast cell line. The addition of enrofloxacin to either doxorubicin or carboplatin resulted in further reductions in osteosarcoma cell viability; this effect was apparent regardless of p53 mutational status or downstream activity.

Effect of preoperative autologous blood donation on patients undergoing bimaxillary orthognathic surgery: a retrospective analysis.

The efficacy of preoperative autologous blood donation (PABD) was evaluated according to preoperative haemoglobin (Hb) values. The records of 295 patients who underwent bimaxillary orthognathic surgery between July 2007 and August 2008 were reviewed. The records for autologous blood donation, intraoperative transfusion, and related laboratory studies were also evaluated. The transfusion trigger used during this period was Hb < 10 g/dl. A total of 189 patients (64.1%) made a PABD and 106 patients (35.9%) did not. The incidence of allogeneic blood transfusion was significantly lower in the PABD group than in the no PABD group (15.9% vs. 29.2%, P = 0.007). This difference was greater in patients with a preoperative Hb < 14 g/dl (20.3% vs. 62.5%, P < 0.0001), and no difference was found in patients with Hb ≥ 14 g/dl (13.3% vs. 14.9%, P = 0.83). PABD reduced the incidence of allogeneic blood transfusion in patients undergoing bimaxillary orthognathic surgery, particularly in patients with a preoperative Hb < 14 g/dl. PABD could be used to reduce the frequency of intraoperative allogeneic blood transfusion in these patients.

Hepatoprotective and neuroprotective tocopherol analogues isolated from the peels of Citrus unshiu Marcovich.

Three tocopherol analogues methoxytocopherol (1), α-tocopherol (2) and γ-tocopherol (3) were isolated from the peels of Citrus unshiu Marcovich. The protective effects of the isolated compounds against tert-butyl hydroperoxide-induced hepatotoxicity in human liver-derived HepG2 cells and glutamate-induced oxidative stress in HT22-immortalised hippocampal cells were evaluated. Compounds 1-3 were significantly protective in HepG2 cells with EC50 values of 21.22 ± 2.01, 25.21 ± 2.11 and 25.25 ± 1.21 µM, respectively, and in HT22 cells, compounds 1-3 had EC50 values of 20.62 ± 1.36, 6.44 ± 1.65 and 9.52 ± 1.54 µM, respectively.

In vitro and Lactation Responses in Mid-lactating Dairy Cows Fed Protected Amino Acids and Fat.

The objective of this study was to evaluate the effect of ruminally protected amino acids (RPAAs) and ruminally protected fat (RPF) supplementation on ruminal fermentation characteristics (in vitro) and milk yield and milk composition (in vivo). Fourteen mid-lactating Holstein dairy cows (mean weight 653±62.59 kg) were divided into two groups according to mean milk yield and number of days of postpartum. The cows were then fed a basal diet during adaptation (2 wk) and experimental diets during the treatment period (6 wk). Dietary treatments were i) a basal diet (control) and ii) basal diet containing 50 g of RPAAs (lysine and methionine, 3:1 ratio) and 50 g of RPF. In rumen fermentation trail (in vitro), RPAAs and RPF supplementation had no influence on the ruminal pH, dry matter digestibility, total volatile fatty acid production and ammonia-N concentration. In feeding trial (in vivo), milk yield (p<0.001), 4% fat corrected milk (p<0.05), milk fat (p<0.05), milk protein (p<0.001), and milk urea nitrogen (p<0.05) were greater in cows fed RPAAs and RPF than the corresponding values in the control group. With an index against as 0%, the rates of decrease in milk yield and milk protein were lower in RPAAs and RPF treated diet than those of basal diet group (p<0.05). In conclusion, diet supplemented with RPAAs and RPF can improve milk yield and milk composition without negatively affecting ruminal functions in Holstein dairy cows at mid-lactating.

Antinociceptive curcuminoid, KMS4034, effects on inflammatory and neuropathic pain likely via modulating TRPV1 in mice.

BACKGROUND: Curcumin, the active ingredient of turmeric (Curcuma longa), has a wide range of beneficial effects including anti-inflammation and analgesia. However, poor bioavailability of curcumin hinders its clinical application. To overcome this limitation, we modified the structure of curcumin and synthesized new derivatives with favourable pharmacokinetic profiles. Recently, curcumin has been shown to have an antagonizing effect on transient receptor potential vanilloid type 1 (TRPV1) ion channels. We investigated the antinociceptive activity of KMS4034 which had the most favourable pharmacokinetics among the tested curcumin derivatives. METHODS: To evaluate the mechanism of the antinociceptive effects of KMS4034, capsaicin (I(CAP))- and heat (I(heat))-induced currents in TRPV1 expressing HEK293 cells were observed after the application of KMS4034. Nociceptive behavioural measurement using the hot-plate test, formalin test, and chronic constriction injury (CCI) model were evaluated in mice. Also, calcitonin gene-related peptide (CGRP) was stained immunohistochemically in the L4/5 dorsal horns in mice with neuropathic pain. RESULTS: I(CAP) (P<0.01) and I(heat) (P<0.05) of TRPV1 were significantly blocked by 10 µM KMS4034. Behaviourally, noticeable antinociceptive effects after 10 mg kg(-1) of KMS4034 treatment were observed in the first (P<0.05) and second phases (P<0.05) of the formalin and hot-plate tests. The mechanical threshold of CCI mice treated with 10 mg kg(-1) KMS4034 was significantly increased compared with control. Immunohistochemical CGRP expression was decreased in the lamina I-II of the lumbar dorsal horns in KMS4034-treated CCI mice compared with the control (P<0.05). CONCLUSIONS: KMS4034 may be an effective analgesic for various pain conditions.

Optimization of ferrioxamine E concentration as effective supplementation for selective isolation of Salmonella enteritidis in egg white.

Utilization of ferrioxamine E (FE) as a sole source of iron distinguishes Salmonella from a number of related species, including Escherichia coli. FE is not able to serve as a source of iron for E. coli or the Proteus-Providencia-Morganella group. This confers a selective advantage on Salmonella Enteritidis in egg white supplemented with FE. The optimum concentration of FE that promoted a selective advantage for Salmonella in egg white was determined. Four supplementation concentrations were evaluated (25, 50, 200, and 500 microg/ml) in egg white artificially inoculated with proportionally mixed cultures of a rifampin-resistant strain of Salmonella Enteritidis (0.1 ml of 102 CFU/ml) and E. coli K-12 (0.1 ml of 10(1) through 10(8) CFU/ml). After a 24-h incubation at 37 degrees C, Salmonella and E. coli populations were enumerated. At higher concentrations of FE (>50 microg/ml), both Salmonella and E. coli were able to use the iron supplement (1 to 8.5 log CFU/ml and 1.8 to 8 log CFU/ml, respectively); however, lower FE concentrations (< or = 50 microg/ml) exclusively promoted Salmonella growth. Salmonella was unrecoverable without supplementation. This study indicates that optimum levels of FE supplementation in egg can improve the selective detection for Salmonella Enteritidis among other competitive organisms.

Synergistic effects of sildenafil on relaxation of rabbit and rat cavernosal smooth muscles when combined with various vasoactive agents.

OBJECTIVE: To evaluate which vasoactive agents have synergistic effects on the cavernosal smooth muscles of rabbits and rats when the agents are combined with sildenafil. MATERIALS AND METHODS: Relaxation responses of cavernosal smooth muscle to single agents (phentolamine, moxisylyte, sodium nitroprusside, forskolin, vasoactive intestinal peptide, VIP, papaverine and sildenafil) in the rabbit, and prostaglandin-E1 and sildenafil in the rat, and to combinations of each agent plus sildenafil, were assessed in vitro. The response to sildenafil of the rabbit strips with and without incubation with l-arginine (1 mmol/L) for 20 min was also evaluated. The effective concentrations for a half-maximal response of single agents and combination solutions were compared. RESULTS: All single agents induced concentration-dependent relaxation of the rabbit and rat cavernosal smooth muscles. There was significant synergism on rabbit cavernosal smooth muscle when the sildenafil was combined with forskolin, sodium nitroprusside, VIP or phentolamine. There was also significant synergism with sildenafil plus prostaglandin-E1 in rat cavernosal muscles. There were no synergistic effects of combinations of sildenafil plus moxisylyte, papaverine or l-arginine. CONCLUSIONS: These results suggest potentially effective combined therapies of sildenafil and intraurethral or intracavernosal prostaglandin-E1, intracavernosal forskolin or VIP, or oral phentolamine for patients with erectile dysfunction who have no success after monotherapy with these agents.

Central mu opioid receptor mechanisms modulate mustard oil-evoked jaw muscle activity.

The injection of the small-fibre excitant and inflammatory irritant mustard oil (MO) into the temporomandibular joint (TMJ) region of rats evokes a sustained and reversible increase in electromyographic (EMG) activity of jaw muscles. The 'rekindling' of this nociceptive reflex by intrathecal (i.t.) administration of the opiate antagonist naloxone and mu but not delta and kappa selective opioid antagonist, suggests that it may be modulated by endogenous opioid inhibitory mechanisms.

Antibacterial activity of S-methyl methanethiosulfinate and S-methyl 2-propene-1-thiosulfinate from Chinese chive toward Escherichia coli O157:H7.

S-Methyl methanethiosufinate (1) and S-methyl 2-propene-1-thiosulfinate (2) were easily seperated from Chinese chive (Allium tuberosum L.) using simple column chromatography. Both compounds showed significant antibacterial activities against E. coli O-157:H7 including spoilage microorganism in food. Structural assignment was based on Mass and NMR-spectroscopic methods.

Pathological findings in cumulative irritation induced by SLS and croton oil in hairless mice.

It is known that the pathological features of acute irritant contact dermatitis are specific according to the irritant. However, in chronic irritant contact dermatitis, it is not clear whether specific patterns exist. To investigate whether the specific pathology of acute irritant contact dermatitis is sustained in chronic irritant contact dermatitis, sodium lauryl sulfate (SLS) and croton oil were applied 3x a week for 2 weeks on the dorsal skin of hairless mice using Finn Chambers. The pathologic changes induced by irritants at various concentrations were evaluated using H&E and Luna's staining, as well as immunohistochemistry for 5-bromo-2-deoxyuridine (BrdU), keratin 6 and loricrin. Our results showed that epidermal hyperplasia and inflammatory infiltration were relatively marked in the groups treated with higher concentrations of irritants. These features were more prominent in the 1% croton oil treated group than in the 0.25% SLS treated group. However, lower concentrations of irritants resulted in very similar histological changes, characterized by epidermal hyperplasia with minimal inflammatory infiltration, irrespective of the chemical. Our results suggest that the histological responses to irritants vary with concentration in cumulative irritation, as in acute irritation, but repetitive mild irritation may evoke common histological changes, characterized by epidermal hyperplasia with minimal inflammatory infiltration, irrespective of the chemical used.

Involvement of GABA(A) receptor in modulation of jaw muscle activity evoked by mustard oil application to the rat temporomandibular joint.

The effect of intrathecal administration of the GABA(A) receptor antagonist bicuculline methylbromide on jaw muscle electromyographic (EMG) activity evoked by mustard oil injection into the rat temporomandibular joint was studied. Bicuculline given prior to mustard oil augmented the EMG activity evoked by mustard oil, and "rekindling" of EMG activity was induced by bicuculline given 30 min after mustard oil. These results suggest that central GABA(A) receptors modulate reflex responses to noxious craniofacial stimuli.

Elimination of early Salmonella enteritidis infection after treatment with competitive-exclusion culture and enrofloxacin in experimentally infected chicks.

The effect of normal avian gut flora (NAGF) and enrofloxacin administration on the early infection of young chicks by Salmonella enteritidis (SE) was determined using day-old White Leghorn chicks. Day- old chicks were divided into two groups, untreated control and NAGF-treated, and then infected with 10(6) cfu of SE per chick by oral gavage. The untreated, infected chicks were further divided into two groups and were either left untreated or medicated with a regimen of 10 mg/kg of enrofloxacin in drinking water daily for 10 d, followed by two doses of NAGF beginning at 10 and 8 wk of age in Trial 1 and Trial 2, respectively. Liver, spleen, and cecum samples were tested for the presence of SE, and immunological responsiveness was investigated up to 12 wk of age. Compared with the untreated group, the cecal colonization of SE was significantly (P < 0.05) decreased in the NAGF-treated group in Trials 1 and 2. No significant differences in organ infection were observed in the NAGF-treated vs. untreated birds. Although a significant effect of the combined treatment of enrofloxacin treatment and NAGF on the early infection was not shown in Trial 1, compared with enrofloxacin only or the untreated group, a significant reduction (P < 0.05) in the number of infected chickens and in the number of SE in the cecal contents was observed at 10 wk of age in Trial 2. The enrofloxacin treatment did not increase opportunistic colonization by SE due to the use of the antibiotic in either trial. The plasma and intestinal immunological responses were not significant at the early age (up to 12 wk) of the birds. The use of enrofloxacin, followed by NAGF, could aid the elimination of SE from young chicks persistently infected at an early age. The combined treatment, compared with enrofloxacin alone, protected chickens from reinfection by 40%.

Postnatal development of central nociceptive mechanisms modulating jaw muscle activity in the rat.

Postnatal changes in the electromyographic (EMG) activity of jaw muscles evoked by mustard oil (MO) application into the rat temporomandibular joint region and the recurrence of increased jaw muscle activities after intravenous injection of naloxone were compared among 4, 6 and 8-week-old rats. In all the groups, MO application increased EMG activity on the ipsilateral side, however, 4-week old rats showed only a small increase in EMG activity on the contralateral side. The EMG activity on the contralateral side increased in an age-dependent manner. The recurrence of increased jaw muscle activity was not induced in 4-week old rats. These results suggest that a neural circuit for generating contralateral responses and mechanisms for central excitation are not established until after 4 weeks postnatally.

Combined effect of antibiotic and competitive exclusion treatment on Salmonella enteritidis fecal shedding in molted laying hens.

Salmonella Enteritidis is an important pathogen for the layer industry, primarily because of its ability to infect hens and ultimately contaminate egg contents. Studies have shown that stress situations, such as flock recycling (induced molting), can increase Salmonella Enteritidis problems in the flock. The present study examined the effect of antibiotic treatment and competitive exclusion (CE) on Salmonella Enteritidis shedding in the period following molt and 14-day feed withdrawal. In two separate trials, 48 birds after molt and feed withdrawal were divided into one group that was treated for 10 days with enrofloxacin in water followed by administration of CE culture and a group that was left untreated. Salmonella Enteritidis shedding was significantly reduced in the antibiotic-CE group. The Salmonella Enteritidis shedding rate was 33 and 25% in untreated birds versus 4 and 0% in the enrofloxacin-CE group on the two test days. These results indicate that treatment of Salmonella Enteritidis-positive laying hens after molting with enrofloxacin and CE culture can substantially reduce Salmonella Enteritidis problems due to molting and would be a possible alternative to diverting eggs for pasteurization or slaughtering the infected flock. Possible development of bacterial resistance in conjunction with antibiotic use is also discussed.

Relation between lymphocyte subpopulations of peripheral blood and immune responses of modified live hog cholera virus vaccine in pigs treated with an ionized alkali mineral complex.

Thirty-nine healthy pigs (28-32 days old) were purchased from a commercial swine farm and housed at swine pens of the College. The animals were vaccinated intramuscularly (1 ml) with an attenuated live hog cholera virus (HCV, LOM strain) and then boostered at 5 weeks after the first vaccination. The animals were divided into 4 experimental groups: 0.05% (w/w) PowerFeel-supplemented diet (T-1, n = 10); 3% (w/w) SuperFeed-supplemented diet (T-2, n = 10); diluted PowerFeel solution (1 : 500, v/v) as drinking water (T-3, n=9); control (n=10). PowerFeel is an original form of ionized alkali mineral complex (IAMC) and SuperFeed is a commercial product of IAMC. The subpopulation of lymphocyte in blood was assayed by a flow cytometry and HCV-specific antibody was determined by an indirect immunofluorescence assay. In IMAC-treated groups, the proportions of subpopulation expressing MHC-class II, CD2+, CD4+, CD8+, and surface IgM+ B lymphocytes were significantly decreased at 5-weeks after the first vaccination. Significant decreases were also observed in the proportions of MHC-class II, CD2+ and CD8+ lymphocyte at 3-weeks after the booster injection. The humoral immune responses in T-1 and T-2 groups were greater than those in T-3 or control group. These results suggest that IAMC-supplemented diets may have an HCV-specific immunostimulatory effect in pigs.

[A case of recurrent squamous cell carcinoma of cervix successfully treated with BOMP].

Combination chemotherapy consisting of bleomycin, vincristine, mitomycin C and cisplatin (BOMP) was applied to a case (57 years old) with recurrence of squamous cell carcinoma of the cervix. The patient with FIGO stage IIIb cervical carcinoma had been treated with radiotherapy seven years ago. She was affected by malignant pericardial effusion, and was treated with direct intrapericardial administration of cisplatin (20 mg). After instillation, ultrasonography and computed tomography showed the wall thickening of colon and marked ascites. Barium enema showed the circumferential narrowing and serrations of colon, and it was diagnosed to be carcinomatous peritonitis. She was treated with BOMP consisting of BLM (5 mg, i.m., days 1 - 5), VCR (1 mg, i.v., day 5'), MMC (8 mg, i.v., day 5), and CDDP (70 mg, d.i.v., day 5). After three courses of BOMP therapy, she achieved a complete response with few toxic effects and did not require special care. She has been free of disease for six months and her performance status is 0. This encouraging result warrants the use of this combination chemotherapy regimen in other patients with recurrent squamous cell carcinoma of the cervix.

[Pharmacokinetic studies on continuous intraportal 5-fluorouracil infusion].

Pharmacokinetics of 5-FU administered by continuous intraportal infusion were studied by infusing 5 mg/kg, 10 mg/kg, 20 mg/kg, and 40 mg/kg of 5-FU, respectively, into the ileocecal vein for an hour. Blood samples were collected from portal vein, femoral vein and liver, and small intestine specimens were obtained at proper intervals. The rabbits were sacrificed at 120 minutes from the start of 5-FU infusion. The results were as follows: Extremely high concentrations of 5-FU in the portal vein, femoral vein and liver tissue were observed in the 5-FU group infused with 40 mg/kg. This phenomenon was suggested to rely on the effect by which the 5-FU catabolic enzyme, dihydrouracil dehydrogenase, was saturated with a large inflow of 5-FU. FdUMP concentration of the liver was lower than that of the small intestine in all groups. These results suggest that a large dose of 5-FU infusion is effective to increase the FdUMP concentration in the liver with continuous intraportal 5-FU infusion.

Selective effect of chronic lead ingestion. II: Effect on phenylethanolamine N-methyltransferase activity in brain regions of rats.

Selectivity of lead effect to phenylethanolamine N-methyltransferase (PNMT) activity in regions of brain from rats postnatally exposed to lead was tested. Three groups of animals were prepared; (1) Rats exposed to lead at a low dose (0.05% PbAcetate: PbAc); (2) Rats exposed to lead at a high dose (0.2% PbAc); (3) Age-matched normal control rats. At 2, 4, 6 and 8 weeks of age weight of whole brain and body in each group were measured. At the same ages activities of PNMT and Na+/K(+)-ATPase were examined on 4 brain regions of each animal. Exposure of rats to lead generally decreased activity of Na+/K(+)-ATPase and showed alternative change of those of PNMT. Brain regions where changes of PNMT activity were detected without concomitant changes of Na+/K(+)-ATPase activity, were telencephalon and pons/medulla at 2 weeks of age and telencephalon at 4 weeks of age in rats exposed to lead at a low dose, and those in rats exposed to lead at a high dose were pons/medulla at 8 weeks of age. These data imply that adrenergic nervous system in the brain regions described above could selectively be affected by lead.