Porcine circovirus type 2 (PCV2) vaccination reduces PCV2 in a PCV2 and Salmonella enterica serovar Choleraesuis coinfection model.

We previously reported that prior porcine circovirus type 2 (PCV2) infection potentiates the severity of clinical signs, lung lesions, and fecal shedding and tissue dissemination of Salmonella enterica serovar Choleraesuis in infected pigs. Here, we evaluated whether PCV2 vaccination is effective in reducing fecal shedding and tissue dissemination of S. Choleraesuis and improving clinical signs associated with PCV2 and S. Choleraesuis infection in 15 Cesarean-derived, colostrum-deprived pigs randomly assigned to 3 groups (n=5/group). The vaccinated and co-infected (VAC-COINF) group received 2 ml of a commercial PCV2 vaccine at age 3 weeks. The VAC-COINF and co-infected (COINF) groups were inoculated intranasally with PCV2 and S. Choleraesuis at 5 and 7 weeks of age, respectively. The CONTROL group pigs received a similar volume of PBS for sham-vaccination and sham-inoculation. PCV2 vaccination clearly reduced PCV2 DNA load in the serum and postmortem tissue samples and decreased PCV2 antigen levels in tissue samples of the VAC-COINF group. After S. Choleraesuis infection, the incidence of several clinical signs increased in the VAC-COINF group compared to that in the COINF group. The microscopic lung lesions and weight gain, fecal shedding and tissue dissemination of S. Choleraesuis except in the spleen were not significantly different in the VAC-COINF and COINF groups. Thus, PCV2 vaccination reduced PCV2 in the S. Choleraesuis and PCV2 coinfection model and the effects on S. Choleraesuis were minimal.

Porcine circovirus type 2 potentiates morbidity of Salmonella enterica serovar Choleraesuis in Cesarean-derived, colostrum-deprived pigs.

PROBLEM ADDRESSED: Porcine circovirus type 2 (PCV2) and Salmonella enterica subspecies enterica serovar Choleraesuis (S. Choleraesuis) are two leading causes of economic loss in the swine industry. Although S. Choleraesuis infection occurs concurrently with PCV2-associated disease in many swine herds, the pathogenesis of concurrent infection with PCV2 and S. Choleraesuis remains largely undefined. OBJECTIVE: We investigated the interactions between PCV2 and S. Choleraesuis in 20 Cesarean-derived, colostrum-deprived (CDCD) pigs randomly assigned to 4 groups (n=5 per group). METHODS AND APPROACH: Pigs in the dual-infected and PCV2-infected groups were inoculated intranasally with PCV2 at 5 weeks of age, and pigs in the dual-infected and S. Choleraesuis-infected groups were inoculated intranasally with S. Choleraesuis at 7 weeks of age. Pigs in the control group served as uninfected controls. RESULTS: After S. Choleraesuis inoculation, severe clinical signs, reduction of weight gain, and severe microscopic lung lesions were observed in dual-infected pigs compared to those in other groups. In addition, the pigs in the dual-infected group shed significantly (P=0.002) higher quantities of S. Choleraesuis in feces 12 days after S. Choleraesuis inoculation, and S. Choleraesuis was recovered from more tissues in this group 14 days after S. Choleraesuis inoculation. CONCLUSIONS: These results indicate that prior PCV2 infection potentiates the severity of clinical signs, lung lesions, and fecal shedding and tissue dissemination of S. Choleraesuis in infected pigs. Therefore, dual infection of pigs with PCV2 and S. Choleraesuis may increase clinical effects of salmonellosis in the field.

Increases in discontinuous rib cartilage and fused carpal bone in rat fetuses exposed to the teratogens, busulfan, acetazolamide, vitamin A, and ketoconazole.

Skeletal changes induced by treatment of pregnant rats with four potent teratogens, busulfan, acetazolamide, vitamin A palmitate, and ketoconazole, were evaluated using Alizarin Red S and Alcian Blue double-staining to investigate the relationship between drug-induced skeletal malformations and cartilaginous changes in the fetuses. Pregnant rats (N = 8/group) were treated once or twice between gestation days (GDs) 10 to 13 with busulfan at doses of 3, 10, or 30 mg/kg; acetazolamide at 200, 400, or 800 mg/kg; vitamin A palmitate at 100,000, 300,000, or 1,000,000 IU/kg; or ketoconazole at doses of 10, 30, or 100 mg/kg. Uterine evaluations and fetal external and skeletal examinations were conducted on GD 20. Marked skeletal abnormalities in ribs and hand/forelimb bones such as absent/ short/bent ribs, fused rib cartilage, absent/fused forepaw phalanx, and misshapen carpal bones were induced at the mid- and high-doses of busulfan and acetazolamide and at the high-dose of vitamin A palmitate and ketoconazole. Increased incidences of discontinuous rib cartilage (DRC) and fused carpal bone (FCB) were observed from the low- or mid-dose in the busulfan and acetazolamide groups, and incidences of FCB were increased from the mid-dose in the vitamin A palmitate and ketoconazole groups. Therefore, DRC and FCB were detected at lower doses than those at which ribs and hand/forelimb malformations were observed in the four potent teratogens.

Shedding of porcine circovirus into colostrum of sows.

The purpose of the present study was to investigate porcine circovirus (PCV) shedding into the milk of sows. Colostrum was collected from 33 sows. PCV1 was isolated from four of 33 milk whey samples. PCV1 DNA was detected by polymerase chain reaction in three of these samples and in three of 10 milk cell samples. PCV2 was also isolated and detected from every single milk whey sample. These results showed that PCV1 and PCV2 were shed into the milk of sows and suggest that PCV can be transmitted to offspring by an oral route through milk.

Experimental reproduction of postweaning multisystemic wasting syndrome in cesarean-derived, colostrum-deprived piglets inoculated with porcine circovirus type 2 (PCV2): investigation of quantitative PCV2 distribution and antibody responses.

Sixteen cesarean-derived, colostrum-deprived piglets were inoculated intranasally with porcine circovirus type 2 (PCV2), originally isolated from a pig affected with postweaning multisystemic wasting syndrome (PMWS). At 1 day postinoculation (PI), 3 of the 5 piglets in the uninoculated control group were moved to the room of inoculated piglets for contact exposure. Porcine circovirus type 2 was detected by polymerase chain reaction (PCR) in swabs from inoculated piglets from 1 day PI and from contact piglets from 2 days after cohabitation. Porcine circovirus type 2 was also detected in all serum samples but not in control piglets 7 days PI. Until the end of study, PCV2 was detected in swabs and serum samples by PCR but not in the control piglets. One inoculated piglet died suddenly without clinical signs 19 days PI. Beginning at 14 days PI, 5 piglets, including 1 contact piglet, had clinical signs of depression, anorexia, and icterus, and 1 inoculated piglet died 21 days PI. Most of the piglets exhibiting the above clinical signs became moribund and were necropsied 21 and 28 days PI. In the piglets that showed clinical signs, gross lesions, including icterus of liver and hemorrhage in stomach, and typical histopathological lesions of PMWS, such as lymphoid depletion and basophilic intracytoplasmic inclusion bodies in lymph nodes and other tissues, were observed. Porcine circovirus type 2 was detected by PCR in all tissue samples except in those of the control piglets. Porcine circovirus type 2 was recovered from several tissue samples of the piglets necropsied until 35 days PI. In particular, PCV2 was recovered in high titer from most of the tissue samples of the piglets exhibiting clinical signs. Serum antibody against PCV2 was mostly detected in inoculated piglets and in contact piglets 14 and 21 days PI by an indirect fluorescence antibody test but was not detected in the piglets exhibiting clinical signs until 28 days PI. These results indicate that PCV2 was able to induce clinical PMWS in the absence of other swine pathogens and that there were significant differences in both the quantitative PCV2 distribution in tissues and the antibody response between the piglets that were infected and developed PMWS and those that were infected but remained healthy.

[The effects of intravenous anesthetics, propofol, fentanyl and ketamine on the excitability of spinal motoneuron in human: an F-wave study].

We have investigated the effects of various intravenous anesthetics, propofol, fentanyl and ketamine on the excitability of spinal motoneuron using an F-wave analysis in a total of 28 patients. All patients were divided randomly into three groups as follows; 2 mg.kg-1 intravenous bolus injection followed by 6 mg.kg-1.h-1 infusion of propofol (P group), 1 mg.kg-1 intravenous bolus injection followed by 1 mg.kg-1.h-1 infusion of ketamine (K group), and 5 micrograms.kg-1 injection of fentanyl (F group). The F-wave was determined after supramaximal electrostimulation of the median nerve in distal point. After establishing stable baseline values, intravenous injection of one of the three anesthetics was applied. The F-wave was recorded 3 minutes after the time of bolus administration. We found a significant (P = 0.018) reduction of the persistence from 77.5 +/- 15.2 to 40.9 +/- 16.8% in the propofol group. On the other hand, no significant changes in F-wave parameters were found in ketamine, or fentanyl group. These results suggested that motoneuron excitability in spinal cord could be inhibited by anesthetic dose of propofol, but not by ketamine or fentanyl.

Inhibitory effect of Epstein-Barr virus activation by Citrus fruits, a cancer chemopreventor.

To search useful compounds in Citrus fruit for cancer chemoprevention, we carried out a primary screening of extracts of fruit peels and seeds from 78 species of the genus Citrus and those from two Fortunella and one Poncirus species, which were closely related to the genus Citrus. These Citrus extracts inhibited the Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol 13-acetate (TPA) as a useful screening method for anti-tumor promoters. Our results indicated that Citrus containing substances may be inhibit susceptibility factors involved in the events leading to the development of cancer.

[Patient-controlled sedation in a patient with respiratory dysfunction for reconstructive surgery of submandibular fracture under local anesthesia].

A 70-year-old male with respiratory dysfunction, chronic obstructive pulmonary disease associated with asthmatic component, received patient-controlled sedation (PCS) using propofol for reconstructive surgery of submandibular fracture under local anesthesia. PCS produced sedation levels between eyelid closure with prompt response to verbal commands and drowsiness. Respiratory disturbance and hemodynamic change were not recognized. The patient was very cooperative, and felt comfortable during the operation. Propofol PCS is an appropriate method for a patient with respiratory dysfunction under local anesthesia.

Expandable metallic stent treatment for malignant colorectal strictures.

Four patients were treated by placement of an expandable metallic stent (two Gianturco Z-stents, two Ultraflex stents) for malignant colorectal strictures. All four patients were able to defecate after stent placement. Stent migration was recognized in one patient. Two patients suffered from tenesmus after stent placement.

[Repeated arterial infusion chemotherapy for inoperable hepatocellular carcinoma using implantable drug delivery system].

We performed a clinical evaluation of repeated arterial infusion chemotherapy using an implantable drug delivery system for 41 patients with inoperable hepatocellular carcinoma (HCC). About half of our patients could not undergo transcatheter arterial embolization (TAE) because of extreme tumor extension and/or accompanying advanced cirrhosis. In most patients we implanted a 5 Fr. catheter non-surgically and connected it to an implanted injection port through a subcutaneous tunnel. The treatment schedule was weekly or biweekly intrahepatic one-shot administration of mitomycin C, adriamycin, 5-fluorouracil and epirubicin. The response rate (CR + PR) was 24.4%. The median survival period was 401.1 days. The 6 month, 1-year and 2-year survival rates were 73%, 48% and 24%, respectively. There were no severe side effects nor complications. The implantable drug delivery system will contribute not only to improved therapeutic efficacy for inoperable HCC but also improve the quality of life for patients.

Ascorbic acid prevents the inhibition of DNA synthesis induced by high glucose concentration in cultured human endothelial cells.

Effects of glucose and ascorbic acid on in vitro collagen and DNA synthesis were evaluated in human umbilical vein endothelial cells. Ascorbic acid significantly potentiated collagen synthesis. Incubation of cells with elevated glucose concentration for 24h significantly reduced [3H]-thymidine uptake (2h). However, the addition of ascorbic acid (0.1 mM) dramatically prevented the inhibition of thymidine uptake. Based on these data, it is suggested that ascorbic acid supplementation in diabetics may prevent or ameliorate diabetic angiopathy.

Characterization of two polysaccharides having activity on the reticuloendothelial system from the root of Glycyrrhiza uralensis.

Two polysaccharides, called glycyrrhizans UA and UB, were isolated from the root of Glycyrrhiza uralensis Fischer. They were homogeneous on electrophoresis and gel chromatography, and showed reticuloendothelial system-potentiating activity in a carbon clearance test. Glycyrrhizan UA is composed of L-arabinose: D-galactose: L-rhamnose: D-galacturonic acid in the molar ratio of 20:14:1:3, and glycyrrhizan UB is composed of L-arabinose: D-galactose: D-glucose: L-rhamnose: D-galacturonic acid in the molar ratio of 12:10:1:10:20, in addition to small amounts of O-acetyl groups and peptide moiety, respectively. About 10% (glycyrrhizan UA) and 35% (glycyrrhizan UB) of the D-galacturonic acid residues exist as the methyl esters. Methylation analysis, carbon-13 nuclear magnetic resonance and periodate oxidation studies indicated their structural features.

Time course of ketone body production in the isolated perfused rat liver in response to various stimuli.

Using a non-recirculating perfusion system, we studied the time course of ketone body output from the isolated rat liver in response to various hormones and changes in pH and redox state. The release of 3-hydroxybutyrate (3-OHB) started to be suppressed within 1 min after the addition of insulin (50 mU/ml) and kept half of the basal level even 10 min after its cessation. The addition of glucagon (0.2 microM) caused an increase in both 3-OHB and acetoacetate (AcAc) outputs from fed livers within 5 min, which reached about 150% of the basal level 10 min after the infusion and maintained a constant level through out the experiment. Growth hormone (2 mu/ml) elicited a slight but significant increase in AcAc output soon after the infusion. Epinephrine (10 microM) also caused a slight increase in both AcAc and 3-OHB outputs 9 min after the infusion and maintained a significant increase even 10 min after stopping infusion. The decrease in pH of the perfusate or the addition of ascorbic acid abruptly suppressed the AcAc production. In summary, the present study clearly demonstrated the direct effects of various hormones on ketogenesis in the liver and the usefulness of a non-recirculating liver perfusion system as a tool for the study of ketogenesis.