Mesh-like vascular changes in copper deficiency-induced rat cardiomyopathy

The pathological effects of copper deficiency (COD) are well known. However, the pathogenesis of cardiomyopathy resulting from COD remains unclear. In this study, aimed to elucidate the pathogenesis of COD-induced cardiomyopathy by examining the morphology of the cardiovascular system in copper-deficient rats using histopathology, immunohistochemistry, and scanning and transmission electron microscopy. Changes detected in the myocardium and interstitium were consistent with those reported for COD. Morphological changes included mesh-like changes in the capillary endothelial cells that appear to be a novel finding in COD-induced cardiomyopathy. These changes are hypothesized to result from abnormal vascular remodeling following damage to the basement membrane and due to the mechanical effects of myocardial contractions. Although cardiomyopathy may be associated with microcirculatory disorders arising from these lesions, further investigations are necessary to demonstrate a causal relationship between the pathogenesis of cardiomyopathy and the contribution of these lesions to disease progression.     

Evaluation of truncated LipL32 expressed by Escherichia coli and Pichia pastoris for serodiagnosis of Leptospira infection in rodents

The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then, truncated LipL32 (tLipL32) was designed to contain the region (87–188th amino acid). The tLipL32 was compared between two recombinant expression hosts Escherichia coli and Pichia pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris (tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E. coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results from flaB gene PCR of kidney samples and WB with whole Leptospira cell. Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32 expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of background reaction.     

Assessment of rabbit spleen size using ultrasonography

Assessment of spleen size using the ultrasonography has become a standard practice in human. However, the assessment is not established method in experimental animals. To establish the index to assess the spleen size using ultrasonography, we measured the cross-section image of rabbit spleen during endotoxin shock. The image of the cross-section was appeared as triangle, and the height of the triangular image was defined as the spleen index. This spleen index showed strong correlation with the spleen weight. In conclusion, this method is suitable for observation of changes in rabbit spleen size and may reduce the number of rabbit in the longitudinal studies.     

Changes in photosynthesis,growth, and sugar content of commercial sugarcane cultivars and Erianthus under flood conditions

Sugarcane (Saccharum spp.) is an economical crop in the tropical and subtropical countries. However, because of global climate change, flooding has become problematic, particularly during the rainy season, in Thailand. We investigated the effects of floods on three commercial sugarcane cultivars, namely NiF8, U-thong 6 (UT6), and U-thong 9 (UT9), as well as Erianthus spp. Growth was assessed using a pot experiment in a glasshouse with two treatments: (1) control and (2) 60 d of flooding followed by 30 d of normal conditions. In comparison with control, during prolonged flooding, Erianthus showed greatly decreased CO₂ assimilation, whereas NiF8, UT6, and UT9 showed slightly declined CO₂ assimilation. Growth in plants subjected to 60 d of flooding was less influenced by floods while sucrose content was not affected except in UT6. During flooding, some roots died, resulting in plants compensating adventitious roots to offset the negative effects of root death and to assist them in maintaining their growth, which appeared from the submerged nodes, with different characteristics for each cultivar. However, 30 d after draining, roots remained damaged, while adventitious roots died, resulting in lesser growth as compared with the control, but it did not significantly affect sucrose content and sugar yield. This study suggests that sugarcane plants need to produce the adventitious roots to compensate their roots’ death during flooding and require time to recover their root system after flooding for obtaining the optimum yield and quality at harvest.     

Study of the Structure–Activity Relationship of an Anti-Dormant Mycobacterial Substance 3-(Phenethylamino)Demethyl(oxy)aaptamine to Create a Probe Molecule for Detecting Its Target Protein

The current tuberculosis treatment regimen is long and complex, and its failure leads to relapse and emergence of drug resistance. One of the major reasons underlying the extended chemotherapeutic regimen is the ability of Mycobacterium tuberculosis to attain a dormant state. Therefore, the identification of new lead compounds with chemical structures different from those of conventional anti-tuberculosis drugs is essential. The compound 3-(phenethylamino)demethyl(oxy)aaptamine (PDOA, 1), isolated from marine sponge of Aaptos sp., is known as an anti-dormant mycobacterial substance, and has been reported to be effective against the drug resistant strains of M. tuberculosis. However, its target protein still remains unclear. This study aims to clarify the structure–activity relationship of 1 using 15 synthetic analogues, in order to prepare a probe molecule for detecting the target protein of 1. We succeeded in creating the compound 15 with a photoaffinity group that retained antimicrobial activity, which proved to be a suitable probe molecule for identifying the target protein of 1.     

Species composition of Modellidae and Cerambycidae (Coleoptera) in a coppice woodland

A continuous sampling of canopy beetles was carried out to determine variations in the abundance, species diversity, richness, and composition of the Mordellidae and Cerambycidae in a coppice woodland. Changes in the abundance and the species richness were monitored at three heights in the forest throughout the season in 1999, using yellow and blue water pan traps. The results showed significant variations in the abundance of Mordellidae among the canopy layers, while little variation was found for Cerambycidae. The abundance, species diversity, and richness were generally greater in summer. The results showed distinct species compositions in both families among layers.     

Effects of acrosomal conditions of frozen-thawed spermatozoa on the results of artificial insemination in Japanese Black cattle

The purposes of this study were to examine the relationship between male artificial insemination (AI) fertility and sperm acrosomal conditions assessed by new and conventional staining techniques and to identify possible reproductive dysfunctions causing low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions in Japanese Black bulls. We investigated individual differences among bulls in the results concerning (1) acrosomal conditions of frozen-thawed spermatozoa as assessed by not merely peanut agglutinin-lectin staining (a conventional staining technique) but also immunostaining of acrosomal tyrosine-phosphorylated proteins (a new staining technique), (2) routine AI using frozen-thawed spermatozoa as assessed by pregnancy diagnosis, (3) in vivo fertilization of frozen-thawed spermatozoa and early development of fertilized eggs as assessed by superovulation/AI-embryo collection tests and (4) in vitro fertilization of frozen-thawed spermatozoa with oocytes. The percentages of frozen-thawed spermatozoa with normal acrosomal conditions assessed by the abovementioned staining techniques were significantly correlated with the conception rates of routine AI, rates of transferable embryos in superovulation/AI-embryo collection tests and in vitro fertilization rates. These results are consistent with new suggestions that the distribution of acrosomal tyrosine-phosphorylated proteins as well as the acrosomal morphology of frozen-thawed spermatozoa are AI fertility-associated markers that are valid for the prediction of AI results and that low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions result from reproductive dysfunctions in the processes between sperm insemination into females and early embryo development, probably failed fertilization of frozen-thawed spermatozoa with oocytes.     

Natriuretic peptide inhibition of intestinal salt absorption in the Japanese eel: physiological significance

Electrophysiological studies in vitro demonstrated the significant inhibition by natriuretic peptides (NP) of short-circuit current across the eel intestine, an important osmoregulatory organ. Inhibitory potencies of several members of the NP family were assessed by voltage-clamp determination of net transepithelial salt absorption measured as the short-circuit current I_sc across the intestine of the freshwater-adapted (FW) and seawater-adapted (SW) Japanese eel (Anguilla japonica); the order of potency of synthetic eel peptides was: amidated atrial natriuretic peptide (ANP-NH₂) > ventricular natriuretic peptide (VNP) > atrial natriuretic peptide (ANP) >> C-type natriuretic peptide (CNP). Neither the order of potency nor the absolute potencies were effected by salinity adaptation. The observed potency sequence suggests that inhibition of intestinal absorption is mediated by A-type guanylyl cyclase-coupled NP receptors. The relatively low sensitivity of the intestinal response to NP compared with circulating NP concentrations suggests a role for intestinal regulation by NP which is independent of systemic delivery from cardiac sources. A novel model, incorporating the known immunohistochemical localization of NP-ergic cells and processes in the epithelial layer of the intestine and the dissipation of the Na⁺ electrochemical gradient along the alimentary tract, is developed in which local secretion of NP (in response to a bolus of food) inhibits salt absorption across the intestine regionally in favor of increased nutrient absorption.