Survival of lactococci during passage through mouse digestive tract

One of the important properties of probiotics is the ability to survive in the intestine. There have been few studies on the probiotic property of lactococci, since they are formally not considered to be natural inhabitants of the intestine. To evaluate lactococci as probiotic bacteria, we investigated their ability to survive during gastric transit by in vitro and in vivo tests. When exposed to an in vitro simulated gastrointestinal environment, such as low pH and bile, only Lactococcus lactis subsp. lactis bv. diacetylactis N7 showed a moderate survival rate among the four strains tested. The tested strains were orally administered to mice, and intestinal passage of the ingested strains was monitored by two methods: antibiotics and PCR. Viable cells of strain N7 were recovered from feces within 24-48 h after administration but not at 72 h. Lactococcus lactis subsp. cremoris ATCC 19257, which had a poor survival rate in vitro test, was also detected at 12 h but not at 24 h. These results indicate that lactococci can reach the mouse intestine alive, but not colonize it. If administered daily, viable strain N7 may exist continuously in the intestine. The effect of strain N7 on intestinal microbial balance and on animal health will be the subject of a further study.     

Establishment of Alb-DsRed2 transgenic rat for liver regeneration research

Because serum albumin is specifically produced by mature hepatocytes, detection system of albumin producing cells could be a valuable tool to visualize liver regeneration or development. We have developed here an albumin enhancer/promoter-driven Alb-DsRed2 Tg rat that expresses DsRed2, having liver-specific reporter gene expression of red fluorescent protein. To study the transdifferentiation of bone marrow cells (BMCs) into albumin producing cells, BMCs from the Alb-DsRed2 Tg rat were injected into rats having acute liver damage caused by 2-acetylaminofluorene plus carbon tetrachloride and chronic liver damage by repeated administration of CCl(4). DsRed2-positive cells were generated in the recipient liver after BMC injection. The number of transdifferentiated DsRed2-positive cells in chronic liver injury model was increased comparing with that in acute injury model. We propose that the Alb-DsRed2 Tg rat is well suited to studying in vivo liver regeneration.     

Electrocardiograms Corresponding to the Development of Myocardial Infarction in Anesthetized WHHLMI Rabbits (Oryctolagus cuniculus), an Animal Model for Familial Hypercholesterolemia

The aim of this study was to determine whether features indicative of myocardial ischemia occur in the electrocardiograms (ECG) in myocardial infarction-prone Watanabe heritable hyperlipidemic (WHHLMI) rabbits, an animal model for human familial hypercholesterolemia. ECG were recorded in 110 anesthetized WHHLMI rabbits (age, 10 to 39 mo) by using unipolar and bipolar limb leads with or without chest leads. We noted the following electrocardiographic changes: T wave inversion (37.4%), ST segment depression (31.8%), deep Q wave (16.3%), reduced R wave amplitude (7.3%), ST segment elevation (2.7%), and high T wave (1.8%). These ECG changes resembled those in human patients with coronary heart disease. Histopathologic examination revealed that the left ventricular wall showed acute myocardial lesions, including loss of cross-striations, vacuolar degeneration, coagulation necrosis of cardiac myocytes, and edema between myofibrils, in addition to chronic myocardial lesions such as myocardial fibrosis. The coronary arteries that caused these ECG changes were severely stenosed due to atherosclerotic lesions. Ischemic ECG changes corresponded to the locations of the myocardial lesions. Normal ECG waveforms were similar between WHHLMI rabbits and humans, in contrast to the large differences between rabbits and mice or rats. In conclusion, ischemic ECG changes in WHHLMI rabbits reflect the location of myocardial lesions, making this model useful for studying coronary heart disease.Abbreviations: CHD, coronary heart disease; ECG, electrocardiogram; WHHL, Watanabe heritable hyperlipidemic; WHHLMI, myocardial infarction-prone Watanabe heritable hyperlipidemicCoronary heart disease (CHD) is prevalent in developed countries, including the United States.^16,24 Although potent compounds (for example, statins to inhibit cholesterol synthesis) have been developed to reduce the public health burden of this disease, CHD remains a leading cause of death, and further efforts are needed to reduce associated morbidity and mortality.²⁵ In evaluating the therapeutic effects of CHD interventions, the electrocardiogram (ECG) is an essential tool for examining myocardial function.³⁹In humans, various ischemic ECG changes occur in association with myocardial ischemia and infarction, such as high T wave, ST segment elevation, emergence of the deep Q wave, reduction of R wave amplitude, resolution of ST segment elevation, and T wave inversion.^21,39 In addition, ST segment depression is a typical change observed with subendocardial ischemia.^2,7In the study of myocardial ischemia, animal models that show ECG waveforms comparable to those of human patients with CHD play an important role. This similarity is important not only for assessing the effects of agents for the treatment of CHD but also for assessing adverse effects of newly developed agents on cardiac function. Although ECG have been used to study myocardial ischemia in several species including pigs, dogs, rabbits, rats, and mice,^{3,9,10,14,18,23} most of these studies used coronary ligation models. These models do not fully reflect the events that occur during myocardial ischemia caused by atherosclerotic coronary stenosis, which is seen typically in patients with CHD.The Watanabe heritable hyperlipidemic (WHHL) rabbit⁴⁰ and the myocardial infarction-prone WHHL (WHHLMI) rabbit³³ are animal models for the study of human myocardial ischemia. WHHLMI rabbits spontaneously develop hypercholesterolemia due to a deficiency of receptors for low-density lipoproteins and manifest severe coronary atherosclerosis and myocardial infarction. Importantly, lipoprotein metabolism in WHHL and WHHLMI rabbits resembles that in humans.^28,30 Using these advantages of the WHHL and WHHLMI models, we and others have been studying the effects of hypocholesterolemic and antiatherosclerotic agents on coronary atherosclerosis.^29,32,34 However, ECG were not examined in these studies. Because the rabbit heart is electrophysiologically similar to the human heart,^27,38 using ECG to monitor myocardial function in the WHHLMI rabbit may be valuable.In the present study, we examined whether ECG changes observed in WHHLMI rabbits reflect myocardial ischemia and whether those changes correspond to ECG features in human patients with CHD.     

Oxidation of 3-C-(2-amino-2-deoxy-D-glucopyranosyl)-1-propene compounds and the structure of 3-C-(2-amino-2-deoxy-D-glucopyranosyl)-1,2-propanediol derivatives for a synthesis of 2,3-didehydro-2,7-dideoxy-N-acetylneuraminic acid

Oxidation of 5-acetamido-4,8-anhydro-1,2,3,5-tetradeoxy-D-glycero-D-ido-non-1-enitol [3-C-(2-amino-2-deoxy-beta-D-glucopyranosyl)-1-propene] was studied to search for preparative routes to aminodeoxy didehydro nonulosonic acid derivatives. Since only moderate chiral induction was observed with osmium tetroxide dihydroxylation as well as with peracid epoxidation, the catalytic asymmetric dihydroxylation conditions were applied to give the stereocontrolled formation of 1,2-propanediol derivatives. The structures of these diastereoisomeric 1,2-propanediol derivatives were determined by X-ray crystallographic analyses. The formation of diastereoisomeric 1,2-propanediols also varied with the nature of 2-substituent on the aminodoexy glycosyl moiety. Thus 5-acetamido-4,8-anhydro-3,5-dideoxy-D-erythro-L-ido-nonitol [(2S)-3-C-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-1,2-propanediol] was obtained predominantly up to 70% from 3-C-(2-acetamido-2-deoxyglycosyl)-1-propene by the use of ADmixbeta reagent. The (2S)-propanediol derivative was transformed in a five-step reaction sequence to 2,3-didehydro-2,7-dideoxy-N-acetylneuraminic acid.     

Detection of 3-hydroxy-3-methylglutaryl-coenzyme A reductase protein Cm-HMGR during fruit development in melon ( Cucumis melo L.)

The fruit size of melon (Cucumis melo L. reticulatus) is determined by the amount of cell proliferation in the pericarp during early fruit development. During this stage, expression and activity of the 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene is required for fruit growth. In this study, we performed a detailed analysis of the correlation between the expression of melon HMGR (Cm-HMGR) protein and cell division in the pericarp. Flow cytometric analysis revealed that the length of the cell division stage was correlated with the fruit size. Western gel blotting and tissue printing illustrated the temporal and spatial accumulation pattern of Cm-HMGR protein during fruit development. The accumulation of Cm-HMGR transiently increased at the beginning of the cell division stage in the pericarp, where active cell division occurred. The amount of Cm-HMGR was correlated with the length of the cell division period. These results strongly suggest that the expression of Cm-HMGR is involved in the determination of melon fruit size by regulating cell division during early fruit development.     

Reconstitution into liposomes of the Na+-dependent transport systems for cyclacillin,an aminopenicillin,from brush border membranes of rat small intestine

Triton X-100 extract from brush border membranes of rat small intestine was recombined with egg phosphatidylcholine liposomes by the freeze-thaw sonication method. The treated liposomes showed a Na⁺-dependent uptake of cyclacillin, which was inhibited by a low concentration of mercuric ions and L-phenylalanylglycine, but not by glycine. These are consistent with the absorption characteristics of the antibiotic in situ and indicate that reconstitution of the Na⁺-dependent active cyclacillin transport system of rat small intestine has been achieved.     

Effect of Flanking Sounds on the Auditory Continuity Illusion

Background

The auditory continuity illusion or the perceptual restoration of a target sound briefly interrupted by an extraneous sound has been shown to depend on masking. However, little is known about factors other than masking.

Methodology/Principal Findings

We examined whether a sequence of flanking transient sounds affects the apparent continuity of a target tone alternated with a bandpass noise at regular intervals. The flanking sounds significantly increased the limit of perceiving apparent continuity in terms of the maximum target level at a fixed noise level, irrespective of the frequency separation between the target and flanking sounds: the flanking sounds enhanced the continuity illusion. This effect was dependent on the temporal relationship between the flanking sounds and noise bursts.

Conclusions/Significance

The spectrotemporal characteristics of the enhancement effect suggest that a mechanism to compensate for exogenous attentional distraction may contribute to the continuity illusion.