Establishment of an Invasive Prostate Cancer Model in Transgenic Rats by Intermittent Testosterone Administration

We have established a transgenic rat for adenocarcinoma of the prostate (TRAP) model that features uniform adenocarcinoma development in prostatic lobes at high incidence within a short experimental period. However, no invasive carcinomas with reactive stroma characteristics similar to those in man were observed. We therefore have focused on a new model for invasive carcinoma of the prostate using TRAP rats. In experiment 1, male TRAP rats in groups 1 and 2 were treated with orchiectomy at day 0 of the experiment. Rats in groups 1–3 underwent testosterone propionate (TP) implantation from weeks 1 to 4 and from weeks 6 to 16. Rats in groups 1 and 3 were given 3,2’-dimethyl-4-aminobiphenyl (DMAB) after TP implantation. The rats of group 4 served as controls. In experiment 2, the rats were divided into three groups, none of which received DMAB or orchiectomy, treated with TP continuously or with the treatment withdrawn once or twice. In experiment 1, invasive adenocarcinomas with abundant collagenous stroma were found in the dorsolateral and anterior prostate, some of which showed perineural space invasion at week 16. The number of invasive carcinoma foci was most frequent in group 3. In experiment 2, invasive adenocarcinoma development in the lateral prostates was correlated with the number of TP administration/withdrawal cycles. In conclusion, our newly established rat model for invasive adenocarcinoma of the prostate could serve as a useful preclinical model for evaluating the in vivo efficacy of preventive and therapeutic agents targeting of the tumor microenvironment.     

Effects of mosapride citrate, metoclopramide hydrochloride, lidocaine hydrochloride, and cisapride citrate on equine gastric emptying, small intestinal and caecal motility

Objective

Although extensive work has been done to elucidate the beneficial and unfavorable effects of gastrointestinal prokinetic agents in humans, little is known on the effects of these agents in horses. In this study, we compared the effects of mosapride, metoclopramide, cisapride, and lidocaine on equine gastric emptying, jejunal and caecal motility and evaluated these agents’ adverse drug reactions (ADRs).

Animals

Seven healthy adult Thoroughbreds.

Procedure

Mosapride 1.0 mg/kg and 2.0 mg/kg, metoclopramide 0.2 mg/kg, and cisapride 1.0 mg/kg were dissolved in 100 mL distilled water for oral administration. Lidocaine 1.3 mg/kg was mixed with 500 mL saline for a 30-min intravenous infusion. Oral administration of 100 mL distilled water was used as control. Gastric emptying was evaluated using ¹³CO₂ breath test, and jejunal and caecal motility was assessed by electrointestinography.

Results

The present study demonstrates that mosapride at doses of 1.0 mg/kg and 2.0 mg/kg facilitates gastric emptying in horses. Improved jejunal motility was observed following administration of mosapride (1.0 mg/kg and 2.0 mg/kg), metoclopramide (0.2 mg/kg), and cisapride (1.0 mg/kg). Similarly, improved caecal motility was observed following administration of mosapride (2.0 mg/kg).

Conclusions and clinical relevance

This study shows that among the prokinetic agents studied here, only mosapride (2.0 mg/kg) promotes jejunal and caecal motility in horses. Considering mosapride ADRs profile, it is believed that this compound is useful in the treatment of diseases associated with decreased GI motility, including postoperative ileus.     

Evaluation of total intravenous anesthesia with propofol or ketamine-medetomidine-propofol combination in horses

Objective-To compare the anesthetic and cardiorespiratory effects of total IV anesthesia with propofol (P-TIVA) or a ketamine-medetomidine-propofol combination (KMP-TIVA) in horses. Design-Randomized experimental trial. Animals-12 horses. Procedure-Horses received medetomidine (0.005 mg/kg [0.002 mg/lb], IV). Anesthesia was induced with midazolam (0.04 mg/kg [0.018 mg/lb], IV) and ketamine (2.5 mg/kg [1.14 mg/lb], IV). All horses received a loading dose of propofol (0.5 mg/kg [0.23 mg/lb], IV), and 6 horses underwent P-TIVA (propofol infusion). Six horses underwent KMP-TIVA (ketamine [1 mg/kg/h {0.45 mg/lb/h}] and medetomidine [0.00125 mg/kg/h {0.0006 mg/lb/h}] infusion; the rate of propofol infusion was adjusted to maintain anesthesia). Arterial blood pressure and heart rate were monitored. Qualities of anesthetic induction, transition to TIVA, and maintenance of and recovery from anesthesia were evaluated. Results-Administration of KMP IV provided satisfactory anesthesia in horses. Compared with the P-TIVA group, the propofol infusion rate was significantly less in horses undergoing KMP-TIVA (0.14 +/- 0.02 mg/kg/min [0.064 +/- 0.009 mg/lb/min] vs 0.22 +/- 0.03 mg/kg/min [0.1 +/- 0.014 mg/lb/min]). In the KMP-TIVA and P-TIVA groups, anesthesia time was 115 +/- 17 minutes and 112 +/- 11 minutes, respectively, and heart rate and arterial blood pressure were maintained within acceptable limits. There was no significant difference in time to standing after cessation of anesthesia between groups. Recovery from KMP-TIVA and P-TIVA was considered good and satisfactory, respectively. Conclusions and Clinical Relevance-In horses, KMP-TIVA and P-TIVA provided clinically useful anesthesia; the ketamine-medetomidine infusion provided a sparing effect on propofol requirement for maintaining anesthesia.     

A Preliminary Study of 13C-Phenylalanine and 13C-Dipeptide Breath Tests in Horses

This study aimed to establish a standard dose and sample collection time for ¹³C phenylalanine and ¹³C-Dipeptide breath test in horses. To evaluate dose-dependent effects, healthy horses received 2.5 mg/kg, 5 mg/kg, and 10 mg/kg ¹³C phenylalanine dissolved in 1 ml/kg distilled water and 1.25 mg/kg, 2.5 mg/kg, and 5 mg/kg ¹³C dipeptide dissolved in 2 ml/ kg distilled water. Tmax was observed during the sample collection time. For ¹³C phenylalanine, the standard deviation of Cmax at 5 mg/kg was lower than that of 10 mg/kg. For ¹³C dipeptide, the standard deviation of Tmax was the lowest at 5 mg/kg. This study revealed that an optimal dose for breath tests with ¹³C phenylalanine and ¹³C dipeptide may be 5 mg/kg in horses.     

Modes of inheritance of two apomixis components,diplospory and parthenogenesis,in Chinese chive (Allium ramosum) revealed by analysis of the segregating population generated by back-crossing between amphimictic and apomictic diploids

To investigate the mode of inheritance of apomixis in Chinese chive, the degrees of diplospory and parthenogenesis were evaluated in F₁ and BC₁ progenies derived from crosses between amphimictic and apomictic diploids (2n = 16, 2x). The F₁ population was generated by crossing three amphimictic diploids 94Mo13, 94Mo49 and 94Mo50 with an apomictic diploid KaD2 and comprised 110 diploids and 773 triploids. All the diploid F₁ plants examined were completely or highly eusporous and completely syngamic. All the triploid F₁ plants examined were highly diplosporous and highly parthenogenetic. KaD2 could not transmit its high level of apomixis via monoploid pollen grains. The BC₁ population, generated by crossing 94Mo49 with apomictic triploids found in the F₁ offspring, exhibited heteroploidy; it comprised haploid, diploid, triploid, tetraploid and various aneuploid individuals. In this generation, clear segregation was observed between diplospory and parthenogenesis. Analysis of the BC₁ population suggests that diplospory and parthenogenesis are each controlled by single dominant genes, D and P, respectively. However, all the BC₁ plants characterized as parthenogenetic were diplosporous. The absence of phenotypically eusporous parthenogenetic plants can be explained by assuming that the presence of diplospory gene is a prerequisite for the parthenogenesis gene expression in Chinese chive.     

Surface plasmon resonance analysis on interactions of food components with a taste epithelial cell model

A new device for evaluating the continuity of taste was developed with the use of surface plasmon resonance (SPR). The model of lingual cells was constructed with liposomes immobilized onto an L1 sensor chip for SPR. Using this device, we classified food components into three categories according to the sensorgram pattern and residual ratio on lipid bilayer. Samples in group A strongly interacted with lipid bilayer, those in group B poorly interacted, and those in group C belong to neither group A nor group B. Sweet proteins and gymnemic acids that prolonged sweet perception were categorized in group A. Almost all the carbohydrates investigated and aspartame, of which the taste perception does not continue, belonged to group B. This device made it possible to detect the interaction with lipid bilayer and dissected the mechanism of taste continuity.     

Stress-induced apoptosis by heat shock, UV and γ-ray irradiation in zebrafish embryos detected by increased caspase activity and whole-mount TUNEL staining

SUMMARY: Environmental stress-induced apoptosis in zebrafish Danio rerio embryos was characterized by assaying caspase-3-like activity and whole-mount terminal deoxynucleotidyl nick-end labeling (TUNEL) staining. Severe stress conditions, such as heat shock at 39°C for 1 h, ultraviolet light at 10–100 mJ/cm² and γ-ray irradiation at 5–20 Gy induced extensive apoptosis in embryos. Apoptotic cells were observed after the bud and 1-somite stages in normal embryos by TUNEL staining, and after stress treatment many TUNEL-positive cells were found in the enveloping and deep cell layers and the larval fin. The caspase-3-like activity increased severalfold during stress-induced apoptosis in a dose-dependent manner. These findings indicate that apoptotic pathways, mediated by caspase-3-like activity, play a major role in zebrafish embryogenesis under stress conditions.     

Inhibition of post-mortem muscle softening following in situ perfusion of protease inhibitors in tilapia

ABSTRACT:     A method of introducing protease inhibitors into fish muscle through the bulbus arteriosus was developed using an in situ perfusion technique. Perfusion efficiency was initially tested using eosin and [³⁵S]-methionine. Visible fluorescence was observed in the gill, liver, intestine and dorsal muscle of the eosin-treated tilapia, and the occurrence of eosin in the blood vessels of the dorsal muscle was confirmed under a fluorescence stereoscopic microscope with ultraviolet light. The radioactivity of [³⁵S]-methionine was taken into the dorsal muscle and liver at a concentration of 7.8 Bq/g and 70.2 Bq/g, respectively, after perfusion with 1000 Bq/mL solution. Using the perfusion technique with four kinds of protease inhibitors dissolved in physiological saline, the type of proteases implicated in the post-mortem muscle softening in tilapia (867 ± 195 g, n  = 10/protease inhibitor) was investigated. After the perfusion of leupeptin (serine and cysteine protease inhibitor), benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-fmk; caspase inhibitor), chymostatin (serine protease inhibitor) and ο -phenanthroline (metalloprotease inhibitor), the breaking strength of the perfused muscle was measured as a parameter of the meat toughness and compared with that of the control fish, which were perfused with physiological saline only. The reduction of breaking strength during storage was inhibited by the perfusion of leupeptin and Z-VAD-fmk.     

Postnatal Development of the Stomach in the Japanese Field Vole, Microtus montebelli

The postnatal development of the stomach in the Japanese field vole, Microtus montebelli , were studied with the light and scanning electron microscopes.
Three kinds of cells constituting the fundic glands, that is, mucous neck cells, parietal cells and chief cells were distinguished clearly by light microscopy to have been differentiated around 14 days old. At 4 days old, the structure of pyloric glands was similar to that of adult stage. The investigation on the development of esophageal sac, fundic stomach and pyloric stomach was made by measuring each area of the vertical section. The fundic stomach grew more rapidly than the esophageal sac and pyloric stomach. The ratio of esophageal sac to fundic stomach and pyloric stomach in the area was about 1.5: 2: 1 at 20 days old.
In the fundic glands, 5-hydroxytriptamine (5-HT)- and somatostatin-immunoreactive cells appeared first at 6 days old and at birth, respectively and in the pyloric glands, gastrin-, 5-HT- and somatostatin-immunoreactive cells appeared at 19th day fetus. Gastrin-immunoreactive cells showed a rapid increase in the pyloric glands after birth. The distribution and frequency of endocrine cells were similar to that of adult at about 20 days old.
The fimbria showed bank-like appearance at the beginning and thereafter they showed the process-like structure. The fimbria in both the appearance and the length at 60 days old was similar to that of adult stage.
As a whole, the stomach of the field vole seemed to became mature at 20 days old in the aspects of gastric endocrine cells and morphology of gastric wall, although the fimbria formation was not accomplished at the same period.     

An alternative menaquinone biosynthetic pathway operating in microorganisms

In microorganisms, menaquinone is an obligatory component of the electron-transfer pathway. It is derived from chorismate by seven enzymes in Escherichia coli. However, a bioinformatic analysis of whole genome sequences has suggested that some microorganisms, including pathogenic species such as Helicobacter pylori and Campylobacter jejuni, do not have orthologs of the men genes, even though they synthesize menaquinone. We deduced the outline of this alternative pathway in a nonpathogenic strain of Streptomyces by bioinformatic screening, gene knockouts, shotgun cloning with isolated mutants, and in vitro studies with recombinant enzymes. As humans and commensal intestinal bacteria, including lactobacilli, lack this pathway, it represents an attractive target for the development of chemotherapeutics.