Iron uptake in a bicarbonate-resistant cell line of tobacco

Abstract

In alkaline soils, plant growth is impaired mainly by high pH and high concentration of bicarbonates. The bicarbonate concentration increases the pH value, and causes deficiency of iron. A bicarbonate-resistant cell line (BR line) of tobacco (Nicotiana tabacum L. cv. Burley21) was selected by adding excess bicarbonate ions (20 mmol L^?1) to the culture medium. The pH of the medium was buffered 8.0 to 8.3. Under these conditions, about 80% of iron in the medium became insoluble. However, under such conditions, the BR line grew well. In this report, we examined some characteristics of the growth and iron uptake in the BR line under iron-deficient (i.e. high pH or no-iron) condition. At pH 5.8, the Fe³⁺ reduction activity was not significantly different between the non-selected line and the BR line. At pH 8.0, however, the Fe³⁺ reduction activity of the BR line was higher than that of the non-selected line. In no-iron condition, the growth of the non-selected line was markedly reduced after 2 weeks, while the BR line was not affected. The content of malic acid in both lines increased with the medium pH, and the content in the BR line was higher than that in the non-selected line. The BR line was able to adapt to the conditions, which restricted iron uptake, such as high bicarbonate concentration, high pH, and low iron conditions. The high ability of Fe³⁺ reduction was maintained at even high pH conditions. Further, the BR line may be able to improve the utilization of iron in the cells.     

Genetic relationships between the dominant genotypes of Phytophthora infestans in Hokkaido, Japan

The genetic characteristics of the dominant genotypes of Phytophthora infestans in Japan (US-1, JP-1, Japanese A1-A, A1-B) were compared. Differences were evident in the peptidase genotype, amplified fragment length polymorphism, and RG57 DNA fingerprints. Almost all of the fingerprint bands for the Japanese genotype A1-B were also present in JP-1 and Japanese A1-A, and few bands were unique to Japanese A1-B. These results suggest that the Japanese A1-B genotype was generated from sexual reproduction involving Japanese A1-A and JP-1 or related genotypes.     

Proportions of melanocyte stimulating hormone-immunoreactive cells in the adenohypophysis of Silky fowl and hyperpigmentation-free cockerels

Alpha‐melanocyte stimulating hormone (α‐MSH) is one of the main regulators for melanocytes, and the adenohypophysis is one of the major tissues for the synthesis of this hormone. The Silky fowl is a characteristic breed of chicken with hyperpigmentation throughout the body. The involvement of the adenohypophysis in the hyperpigmentation of this breed is not known. In the present study, the proportion of melanocyte stimulating hormone‐immunopositive cells (MSH cells) in the adenohypophysis was immunocytochemically compared between Silky, Red Cornish × New Hampshire (RN) crossbred and Japanese bantam cockerels at 15 weeks of age. After the body and gland were weighed, the adenohypophyseal cells were enzymatically dispersed and immunostained for α‐MSH, and the immunopositive MSH cells were counted. The weights of the body and adenohypophysis were heaviest in the RN crossbred, followed by the Silky and lightest in the bantam cockerels. In contrast, the ratio between adenohypophysis and bodyweight was much larger in the Silky and the bantam than in the RN crossbreed (P < 0.05). The population of MSH cells in the adenohypophysis was larger in the Silky (14.3%) than in the RN crossbreed (8.0%) and the bantam (8.1%) cockerels (P < 0.05). From these results, it was concluded that prepubertal Silky cockerel have numerous MSH cells in the adenohypophysis suggesting a relationship to hyperpigmentation.     

Spontaneous pinealoma in a male Crj:CD (SD) IGS rat

A pinealoma (benign) was found in a 61-week-old male Crj:CD (SD) IGS rat. The neoplasm was located between the cerebral hemispheres and the cerebellum. Histologically, the tumor cells consisted of two cell types: large, pale-staining cells and small dark-staining cells. A fibrovascular stroma divided the tumor cells into incomplete lobules or nest structures. Relatively numerous mitoses were noted in the tumor cells. Ultrastructurally, the tumor cells contained dense-cored vesicles, approximately 120 nm in diameter.     

Matrix-metalloproteinases-2 and -9 production in bovine endometrial cell culture

In vitro cell culture is a convenient tool for studying cellular mechanisms. In the present study, production of matrix-metalloproteinases (MMPs) in bovine endometrial (containing both epithelial and stromal cells) monolayer cells was examined. Blastocysts attached to the endometrial cells in a monolayer culture were examined for their effects on MMP-2 production. Initial attachment of blastocysts to the monolayer inhibited MMP-2 production by endometrial cells. But once trophoblast cells began to migrate into the endometrial cell layer, MMP-2 production increased, and at the same time MMP-9 production also became evident in the medium. In order to understand how blastocysts affected MMP-2 production, we examined the effect of progesterone, estradiol, insulin-like growth factors (IGFs), tumor necrosis factors (TNFs), and interferon-tau (IFN-tau) supplementation. It was IFN-tau that inhibited the production of MMP-2. In addition, progesterone at a lower dose appeared to inhibit MMP-2 production. Both TNF-alpha and TNF-beta strongly stimulated the production of MMP-2 and MMP-9, whereas IGFs had no effect. Based on these findings, it appears that conceptus has the capacity to inhibit MMP activity.     

Trophinin is expressed in the porcine endometrium during the estrous cycle

We investigated endometrial expression of trophinin mRNA and protein, homophilic cell adhesion molecules, during the estrous cycle of gilts. An immunopositive reaction for trophinin was observed in the luminal and glandular epithelia of the endometrium at all stages of the estrous cycle, but not in endometrial stromal cells or the myometrium. A partial coding sequence of porcine trophinin was similar to sequences in humans and mice, with homologies of 75% and 70%, respectively. As in humans and mice, the trophinin gene is expressed in the endometrium. Trophinin, however, is expressed in the endometrium of the pig throughout the estrous cycle, higher expression levels were observed at some points of the luteal phase, as in humans. These findings suggest that regulation of trophinin gene expression in the pig is different from that in mice, but similar to that in humans. Furthermore, the present results suggest that the pig might be a suitable model for studying the physiological importance of trophinin in early pregnancy in humans.     

Lactoferrin concentrations in milk from normal and subclinical mastitic cows

The concentrations of lactoferrin (Lf) in quarter milk from normal lactating cows and subclinical mastitic cows were measured to determine whether the Lf concentration in milk is influenced by the age of the cow, the stage of lactation, number of milk somatic cells and the presence of pathogens. Lf concentrations in 111 quarter milk samples from 28 normal lactating cows and 270 quarter milk samples from 198 subclinical mastitic cows were measured by means of a single radial immunodiffusion test. Lf concentrations (means +/- standard deviations; logarithmic form) in normal cows and subclinical mastitic cows were 2.23 +/- 0.39 and 2.70 +/- 0.39, respectively. The mean milk Lf concentration (log) in subclinical mastitic cows was significantly (p<0.01) higher than that in normal cows. The mean milk Lf concentration (log) in normal lactating cows aged 5 years was lower than those in normal lactating cows aged 2 years (p<0.01) and 3 years (p<0.05). The results showed that the milk Lf concentration (log) is associated with age of the dairy cow (one-way analysis of variance test, p<0.01). The mean milk Lf concentration (log) in the latter lactational period tended to be higher than those in the peak and middle periods. Milk Lf concentrations (log) tended to be proportional to the level of the somatic cell count (SCC) score. Mean milk Lf concentrations (log) in subclinical mastitic cows infected with Staphylococcus aureus and with other streptococci species were significantly (p<0.01) higher than those in cows infected with coagulase-negative staphylococci and with Corynebacterium bovis.     

Comparison of extracapillary and endocapillary blood flow oxygenators for open heart surgery in dogs: efficiency of gas exchange and platelet conservation

The goal of the current study was to compare the efficiency of gas exchange and platelet conservation of a new extracapillary blood flow oxygenator versus an endocapillary blood flow oxygenator during open heart surgery with extracorporeal circulation in dogs. Dilation and remodeling of the right ventricular outflow tract of dogs was performed using a patch graft technique to simulate pulmonary stenosis. Sequential pre- and post-operative blood analysis revealed that gas exchange efficiency and platelet conservation was significantly greater with the extracapillary blood flow oxygenator than with the endocapillary blood flow oxygenator. However, the priming volume of the extracapillary blood flow oxygenator was significantly greater, leading to hemodilution. We conclude that while the extracapillary blood flow oxygenator provided benefits in terms of gas exchange and platelet conservation, development of a smaller extracapillary blood flow type oxygenator to reduce hemodilution effects would be beneficial.     

Development of a PCR test for the identification of Staphylococcus intermedius based on the 16S rDNA sequence

The development of a PCR assay based on the 16S ribosomal RNA gene (rDNA) sequence was carried out for the identification of Staphylococcus intermedius. Sixty-six strains of S. intermedius, 70 of Staphylococcus aureus and 2 of Staphylococcus hyicus were examined for the assay. The 16S rDNA, of which the PCR target fragment makes up 901 bp corresponding to the sequence data of the gene, was detected in all strains of S. intermedius, but it was not detected in any strains of either S. aureus or S. hyicus. These results suggest that the PCR allows a simple and precise identification of S. intermedius.     

Experimental and clinical studies on plasma leptin in obese dogs

Leptin is a protein synthesized and secreted primarily by adipocytes, and the circulating leptin concentration is elevated in obese humans and rodents. Recently, we have established a sandwich enzyme-linked immunosorbent assay for canine leptin. In the present study, plasma leptin concentrations were measured in experimentally developed obese beagles and in clinically obese dogs. When 5 male beagles were given a high-energy diet for 3 months, all of them became obese and the plasma leptin concentration significantly increased from 2.4+/-1.2 to 4.9+/-0.9 ng/ml, positively correlating with body fat content estimated by the deuterium oxide dilution method (r=0.87). The leptin concentrations of plasma samples collected from 59 dogs in veterinary practices were compared with their body condition scores (BCS). The plasma leptin concentrations of obese dogs were 9.7+/-0.7 and 12.3+/-1.5 ng/ml at BCS=4 and BCS=5, respectively, which were significantly higher than those of optimal (BCS=3) dogs (2.7+/-0.3 ng/ml). There was no significant effect of sex and breed. A weak positive correlation (r=0.37) was found between the plasma leptin concentration and age, probably due to the lesser content of visceral fat in puppies younger than 1 year old. These results indicate that plasma leptin is a good index of adiposity in dogs regardless of breed, age and sex, and may be useful for quantitative assessment of obesity in small animal practice.