Genome-wide association QTL mapping for teat number in a purebred population of Duroc pigs.

Because of increasing litter size in Western pig breeds, additional teats are desirable to increase the capacity for nursing offspring. We applied genome-wide SNP markers to detect QTL regions that affect teat number in a Duroc population. We phenotyped 1024 animals for total teat number. A total of 36 588 SNPs on autosomes were used in the analysis. The estimated heritability for teat number was 0.34 ± 0.05 on the basis of a genomic relationship matrix constructed from all SNP markers. Using a BayesC method, we identified a total of 18 QTL regions that affected teat number in Duroc pigs; 9 of the 18 regions were newly detected.

A new expression of pharmacological activity, ET63.

We studied the response time of drugs in relation to the feedback mechanism in animals. The time from drug administration to onset of loss and recovery of righting reflex or death by hexobarbital in mice were measured at several dose levels and it was demonstrate that in every case the time (t)- response rate (R) function could be expressed as: (formula: see text) These parameters could be obtain by use of only a small number of animals at every dose level. The result suggests that the pharmacological activity of not only pure drugs but also of commercial chemicals with fixed standard of purity and concentrations may be expressed by the parameter t63 transposed as ET63. t63: time when cummulative response rate reaches 63%. t1: latency.

[Pharmacological actions of iprazochrome on the vascular system].

The pharmacological actions of iprazochrome (IC) on the vascular system were studied, and the following results were obtained: No death nor abnormal behaviors were observed in acute toxicity tests conducted on male and female mice and rats despite the administration of large doses of IC (10,000 mg/kg, p.o. and 80 mg/kg, i.v., respectively). IC inhibited dose-dependently platelet aggregation in vitro induced by arachidonate and ADP, whereas no effect was observed on ADP-induced respiratory depression in mice, which is closely related to platelet aggregation in vivo. The antiserotonergic actions of IC on the isolated external carotid arteries and femoral arteries in dogs observed in a noncompetitive manner were found to be 1/24 to 1/65 that of methysergide. On the other hand, IC showed no inhibitory effect on the paw edema of rats in vivo induced by serotonin. The inhibitory effect of IC on peritoneal dye leakage in mice was less than half that of phenylbutazone. IC prevented apoplexy in stroke-prone SHR (SHRSP) without lowering the blood pressure. Histological changes in the cerebrum of SHRSP were ischemic changes such as swelling of the neurons and shrinkage of the nuclei, mainly in the cerebral cortex and corpus striatum area.

[Studies on anti-inflammatory agents. (5). Specific characteristic of bone changes in adjuvant arthritic rats with passage of time].

Adjuvant arthritic rats develop marked changes in the distal limb joints. These changes were observed by soft X-ray photography at appropriate intervals. It was confirmed that the "cotton wool appearance" surrounding the bones, which was readily discernible from the radiographs, was a good indicator for following the progression of bone damages. Standard radiographs were selected to represent the various bone changes based on the "cotton wool appearance". When the incidence rates of bone deformation confirmed on the basis of the above method were plotted against Weibull Probability Paper, the plots conformed with the Weibull distribution function. Thus, it was possible to evaluate bone changes in adjuvant arthritic rats by observing the "cotton wool appearance" surrounding the bones.

[Experimental studies on gastric ulcer (6). Endoscopical evaluation of healing processes of acetic acid ulcer in rats (2): Healing, recurrence and relapse of ulcer].

The progress of acetic acid-induced gastric ulcers in rats were sequentially observed with an endoscope for 365 days, and the influences of the ulcer-induced site, age and sex on the healing, recurrence and relapse of these ulcers were investigated. Submucosal injections of 20% acetic acid at the region between the fundus and the pylorus on the anterior wall of the stomach (site A) in 7 weeks old male rats produced active round-shaped ulcers with blood coagulation and debris on the 3rd day after ulcer induction. The ulcers diminished in size with the progress of time (10--50 days), and 50% of the ulcerated rats healed with convergences of mucosal folds (35--154 days). On and after the 50th day, some diminished or healed ulcers showed signs of relapse (37% of used rats) or recurrence (40% of healed rats, 21% of used rats), with the cumulative relapse and recurrence percentage (CR%) reaching 59% on the 365th day. Ulcers in the glandular portion on the greater curvature (site B) healed significantly faster than those in site A. All of the ulcers in site B healed within 133 days and did not recur or relapse. The cumulative healing percentage (CH%) and the CR% of the ulcers in site A in 25 weeks old male rats were 33% and 67%, respectively, and those of the ulcers in site A in 7 weeks old female rats were 60% and 60%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

[Experimental studies on gastric ulcer (5). Endoscopical evaluation of healing processes of acetic acid ulcer in rats (1). Ulcer reducing process].

The following results were obtained as the result of the sequential observations with an endoscope of the gastric ulcer produced by submucosal injection of acetic acid in rats. 1) The size of the ulcer produced by acetic acid injection observed on the 3rd day after ulcer induction depended on the concentration and volume of acetic acid. 2) The natural reducing rates of the size of ulcers in the region between the fundus and pylorus on the anterior wall (A) and in the glandular region on the greater curvature (B) were compared. The reducing rate of B ulcer was significantly faster than that of A ulcer. 3) No significant difference was observed in the reducing processes of untreated ulcers in 7 weeks old rats and 25 weeks old rats. 4) The ulcer reducing rate of female rats was found to be slightly faster than that of male rats. 5) The ulcer reducing rates of the aldioxa (ALD) and sucralfate (SUC) treated group were significantly faster than that of the control group. However, no difference in the ulcer reducing process between the cimetidine (CIM) treated group and the control group was observed. 6) The ulcer indices (UI) of both the combinations of ALD and SUC and of ALD and CIM were found to be significantly less than that of the control group on the 20th day. Treatment with drug combinations shortened the healing period of ulcers more than treatment with one drug alone. The percent healing by the combination of ALD and SUC was the highest among the groups treated. 7) An approximately linear relationship exists between the logarithm of the days after ulcer induction (x) and logarithm of UI (y), and the following equation was obtained: log y = a log x + b. Slope (a) indicates the ulcer reducing rate in evaluating the ulcer reducing process.

Experimental studies on gastric ulcer. (4). Sequential observation and evaluation of gastric ulcers by endoscope in the rat.

In the present study, a sequential observation of the gastric lesions or ulcers in rats weighing from 100 g was made with an endoscope. It was found that bleeding, edema, hyperemia, fur of ulcer and convergences of the mucosal folds in the stomach could be observed in rats with a SES-1717S (phi = 1.7 mm) or SES-2217S (phi = 2.2 mm) endoscope. Furthermore, we have devised a method which we named the "sohmen method" for measuring ulcer-sizes with an endoscope. The ulcer-size was compared with 5 mm lengths of sohmen (Japanease vermicelli) which were inserted through the sheath of the endoscope into the stomach. Therefore, the ulcer-size could be accurately measured by this method. By this observation method, it is possible to observe repeatedly over a long term the changes in gastric lesions or ulcers, to measure changes in ulcer-sizes and to evaluate the relapse or recurrence of ulcers in the same rats.

[Experimental studies on antitumor effects of lysozyme-II. Effect of lysozyme on immunopotentiation].

Effects of lysozyme (LY) on immuno-responses were examined. Tumor-bearing mice were used on most of experiments and normal mice on several cases of experiments. Cytostatic activity on the Peyer's patch cells in Meth-A bearing Balb/c mice was enhanced at 6 to 12 hrs after oral administration of LY. Furthermore, LY showed to recover from the decline of blastogenic activity, which was observed on the lymph node cells and the spleen cells of tumor-bearing mice, at 8 and 14 days after tumor inoculation. In the normal ICR mice, it was not observed that LY increased the foot-pad reaction against the sheep red blood cells (SRBC). On the other hand, the decrease of the foot-pad reaction, which was observed in the tumor bearing mice at 20 days after Sarcoma-180 (S-180) inoculation, was recovered by the administration of LY. A decrease of the number of plaque-forming cells (PFC) against SRBC which was induced with tumor inoculation in the spleen cells was not observed at 20 days after S-180 inoculation, but LY caused a significant increase of PFC response in spleens on both of normal and tumor bearing mice. From these results, it was suggested that potentiation of systemic immuno-response derived from the activation of Peyer's patches brought about antitumor effects of LY.

Inhibitory mechanism of dipyridamole on platelet aggregation ex vivo.

The effects of dipyridamole on platelet aggregation ex vivo and in vitro and on platelet cyclic AMP phosphodiesterase (PDE) were studied, and the mechanism of the ex vivo effects was assessed. Both the ADP- and collagen-induced aggregations ex vivo were inhibited dose-responsively by oral administration of dipyridamole. Maximum dipyridamole levels in the plasma were reached at 30 min after the administration. The inhibitory effects of dipyridamole on platelet aggregation ex vivo reached a maximum at between 1 and 2 hrs. On the other hand, the ADP-induced aggregation in vitro and cyclic AMP PDE activity were not inhibited until after 10 min of incubation at a low concentration of dipyridamole. This mode of inhibition of platelet aggregation in vitro and of cyclic AMP PDE activity agreed with the mode of inhibition in the case of platelet aggregation ex vivo. It is suggested therefore that the ex vivo effects, observed with only a low dipyridamole concentration in the plasma, may be due primarily to inhibition by dipyridamole of the cyclic AMP PDE in platelets.

[Experimental studies on antitumor effects of lysozyme].

Effects of lysozyme(LY) alone and combination with various antitumor agents were examined using syngeneic tumors mainly Meth-A and MH 134 transplanted into Balb/c and C3H/He mice, respectively. The effects of LY on pulmonary metastases were also examined in BDF1 mice bearing Lewis lung carcinoma (3LL). LY inhibited the growth of Meth-A tumor and enhanced antitumor effects of mitomycin C (MMC). bleomycin (BLM) and 5-fluorouracil (5-FU), LY activated antitumor effects of MMC, BLM and 5-FU on MH 134 tumor, however, LY alone did not show any significant antitumor effect on it. The combination treatments using LY with MMC or 5-FU showed a marked inhibition on pulmonary metastases of 3LL. The pre-treatment of LY inhibited the tumor growth of Sarcoma-180 in ICR mice.

[Effects of dihydroergotamine mesylate (DEM) on the cardiovascular and nervous systems (author's transl)].

Effects of DEM on the cardiovascular, autonomic, and central nervous systems were studied using dogs, rats and rabbits. DEM showed relatively weak effects on the EEG in curarized rabbits and on ambulatory and drinking activities in rats. DEM suppressed dose-dependently the pressor responses induced by norepinephrine, serotonin and bilateral carotid occlusion, but had no effect on the depressor responses induced by acetylcholine and electrical stimulation to the postganglionic vagus nerve in anesthetized dogs. DEM produced a dose-dependent increase in blood pressure, a dose-dependent decrease in heart rate, a transient decrease in respiration rate, and little change in the ECG of anesthetized dogs. The pressor responses induced by DEM were inhibited by phentolamine, methysergide, and indomethacin in anesthetized dogs. With canine femoral arteries, DEM-induced contractions were blocked 10% by phentolamine, 60% by methysergide, and enhanced 10% by indomethacin. On the other hand, DEM-induced contractions of the femoral veins were blocked 30% by phentolamine, 60% by indomethacin, and 10% by methysergide. DEM provoked a significant increase in the level of prostaglandin E in the bathing fluid of the veins but not in that of the arteries. The results suggest that the arterioconstrictor responses induced by DEM are mediated mainly through the serotonin receptors, and the venoconstrictor responses induced by DEM mainly through the enhanced synthesis of prostaglandin E.

Studies on an anti-inflammatory agent. III. Pharmacological investigations of a new non-steroidal anti-inflammatory agent: 2-oxo-3-[4-(1-oxo-2-isoindolinyl)-phenyl]-butanamide (GP 650).

The anti-inflammatory, analgesic, antipyretic and ulcerogenic activities and acute toxicity of 2-oxo-3-[4-(1-oxo-2-isoindolinyl)-phenyl]-butanamide (GP 650) were investigated in laboratory animals and compared with those of phenylbutazone, acetylsalicylic acid (ASA), ibuprofen, tiaramide-HCl and tinoridine-HCl. The anti-inflammatory and analgesic activities of GP 650 were found to be almost equivalent to those of phenylbutazone on the basis of various anti-inflammatory and analgesic tests while its antipyretic activity was less marked, and its ulcerogenic action and acute toxicity were much lower than those of the other agents. It is interesting to note that GP 650 possesses marked anti-inflammatory activity similar to acidic anti-inflammatory agents in chronic inflammatory models along with mild antipyretic and ulcerogenic activities found in basic anti-inflammatory agents. Therefore, GP 650 appears to be a promising anti-inflammatory and analgesic agent.

[Comparative responsiveness of isolated saphenous, femoral and external carotid arteries and veins of dogs to dihydroergotamine mesylate (DEM) (author's transl)].

Changes in tension of spiral strips of saphenous, femoral and external carotid arteries and veins were measured isometrically. DEM stimulated 6 preparations in almost the same concentration ranges (pD2 values were 8.64 to 8.40). However, the dose-response curves for DEM indicted variations in responsiveness among the different arteries and veins. Compared with norepinephrine (NE) (1)( the intrinsic activity of DEM was 0.002 on saphenous arteries, 0.05 on saphenous veins, 0.03 on femoral arteries, 0.18 on femoral veins, 0.13 on external carotid arteries and 0.13 on external carotid jugular veins. Thus DEM contracted more potently the venous strips from the hind limbs. IN femoral and external carotid arteries, antagonism of serotonin by DEM or methysergide was investigated. DEM displaced the dose-contractile response curves for serotonin in a noncompetitive manner, and the antagonistic response of DEM to serotonin was about 8 times more effective in external carotid arteries (pD'2 value=6.96) than in femoral arteries (pD'2 value=6.05). Methysergide, unlike DEM, antagonized the response to serotonin in a competitive manner at low doses but in a noncompetitive manner in high doses, and was fairly equal in antagonizing response to serotonin in external carotid arteries (pA2 value=that the therapeutic value of DEM in the treatment of orthostatic hypotension is due to its selective anti-serotonin activity on the external carotid arteries.

Studies on the synthesis of chemotherapeutics. 10. Synthesis and antitumor activity of N-acyl- and N-(alkoxycarbonyl)-5-fluorouracil derivatives.

A number of N-acyl and N-(alkoxycarbonyl)-5-fluorouracil derivatives possessing, for example, benzoyl, o-toluyl, acetyl, propionyl, heptanoyl, ethoxycarbonyl, phenoxycarbonyl, and benzyloxycarbonyl groups as N1 and/or N3 substituents were synthesized, and their antitumor activities were evaluated. The synthesis was achieved by a direct and two-step acylation of 5-fluorouracil and by selective N1-deacetylation of N1-acetyl-N3-substituted-5-fluorouracil under appropriate reaction conditions. Several N3-benzoyl- and N3-o-toluyl-5-fluorouracil derivates and showed significant activity against experimental tumor, and N1-acetyl-N3-o-toluyl-5-fluorouracil was found to be most promising among them. Further investigation revealed 12 to retain higher activity toward various tumors, with lower toxicity and good blood level, than either 1 or FT-207, even for oral administration.

A method for evaluating analgesic agents in rats.

A method is described for evaluating antinociceptive activity in rats using writhing responses induced by 4% sodium chloride solution. Narcotic and nonnarcotic analgesics, narcotic antagonists except naloxone, and antipyretic and nonsteroidal antiinflammatory drugs were effectively evaluated at relatively low doses using this method. Parenteral ED50 values of these compounds ran parallel to their clinical doses. The method detected not only the analgesic action of the above drugs but also that of methamphetamine. In spite of its high sensitivity, the method was specific in the sense that CNS-depressant, muscle relaxant, anticholinergic, and antihistamine drugs and anesthetics were inactive even at toxic doses. On the other hand, the decrease in the analgesic activity of morphine upon chronic injection was also observed with this method. The writhing test with 4% sodium chloride in rats was concluded to be a sensitive and specific test for analgesic drugs and useful for examining changes in analgesic activity of drugs during chronic administration.

Influence of adjuvant arthritis on main urinary metabolites of prostaglandin F and E in rats.

The influence of adjuvant arthritis in rats on main urinary metabolites (MUM) of prostaglandin (PG) F and E type was studied. 1) In our model rats, urinary excretion of PGE-MUM increased on day 11 prior to the appearance of secondary lesions, but that of PGF-MUM did not change significantly during the observed period (21 days). The excretion of PGE-MUM was not proportional to the increase in both hind paws volume. 2) Indomethacin, which diminished primary lesions but did not suppress secondary lesions, reduced the increase in urinary excretion of PGE-MUM. 3) Prednisolone and azathiopurine, which suppressed both primary and secondary lesions, increased the excretion of PGE-MUM over adjuvant control values on days 4 and 8. The facts described above suggest that the increase of endogenous PGE during days 4 to 8 may be important in the suppression of secondary lesions in adjuvant arthritis in rats.