Effects of Alendronate on A Disintegrin and Metalloproteinase with Thrombospondin Motifs Expression in the Developing Epiphyseal Cartilage in Rats

A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) have been reported to play a role in the degradation of aggrecan, a major component of cartilage. This study was performed to examine the effects of alendronate on the expression of ADAMTS in developing femoral epiphyseal cartilage. Primary cultured chondrocytes from this cartilage were treated with alendronate in vitro and postnatal day 1 rats were injected subcutaneously with alendronate (1 mg/kg) every second day in vivo . The number of cultured chondrocytes and their aggrecan mRNA levels were unaffected by the alendronate treatment at 10⁻⁶ to 10⁻⁴  m concentrations. The mRNA levels of ADAMTS-1, -2 and -9 in chondrocytes were also unaffected. However, the levels of ADAMTS-5 and -4 were reduced significantly by the same treatment. The thickness of the proliferating chondrocyte layers and the aggrecan mRNA levels in the epiphysis were unaffected by the alendronate treatment in vivo . However, the hypertrophied chondrocyte layers became significantly thicker, and the size of the secondary ossification centre was reduced significantly by the same treatment ( P  < 0.05). Both ADAMTS-4 and -5 mRNA expressions were also reduced significantly in vivo . The immunoreactivity against ADAMTS-4 was seen in hypertrophied chondrocytes and reduced significantly by the alendronate treatment. These results suggested that alendronate can inhibit the degradation of aggrecan in the articular cartilage by downregulating the expression of matrix enzymes such as ADAMTS-4 and -5.     

Measurement of transepidermal water loss from clipped and unclipped anatomical sites on the dog

Objectives   To establish a standardised clipping method for the measurement of transepidermal water loss (TEWL) with a VapoMeter® in Beagle dogs and to identify the optimal anatomical site for TEWL measurement.
Procedure   TEWL values obtained from skin sites on five healthy Beagles clipped using two different blade angles (standard vs non-standard) were compared. TEWL values for 48 h were also obtained from seven different anatomical sites that had differing hair density. The hair was clipped in the intensively haired anatomical sites (head, lower and upper back and tail), but not clipped in the sparsely haired sites (ear, inguinal region, footpad).
Results   The TEWL values for the standard and non-standard clipping sites were 6.3 ± 1.31 and 27.2 ± 1.11 g/h/m², respectively. We found the upper back among the clipped sites was the most appropriate site for TEWL measurement over 48 h after clipping, whereas among the unclipped sites the ear was the most appropriate, because the TEWL values from those anatomical sites had the least fluctuation and were less affected by movement.
Conclusion   The clipping method and anatomical site should be standardised in order to minimise the experimental variation in TEWL measurement in dogs.     

Identification of isolate‐specific resistance QTLs to phytophthora root rot using an intraspecific recombinant inbred line population of pepper (Capsicum annuum)

Quantitative trait loci (QTL) for resistance to phytophthora root rot caused by Phytophthora capsici were investigated using two Korean P. capsici isolates and 126 F₈ recombinant inbred lines derived from a cross of Capsicum annuum line YCM334 (resistant parent) and local cv. Tean (susceptible parent). The experimental design was a split plot with two replications. Highly significant effects of pathogen isolate, plant genotype, and genotype × isolate were detected. QTL mapping was performed using a genetic linkage map covering 1486·6 cM of the pepper genome, and consisted of 249 markers including 136 AFLPs (Amplified Fragment Length Polymorphisms), 112 SSRs (Simple Sequence Repeats) and one CAPS (Cleaved Amplified Polymorphic Sequence). Fifteen QTLs were detected on chromosomes 5 (P5), 10 (P10), 11 (P11), Pb and Pc using two data processing methods: percentage of wilted plants (PWP) and relative area under the disease progress curves (RAUDPC). The phenotypic variation explained by each QTL (R²) ranged from 6·0% to 48·2%. Seven QTLs were common to resistance for the two isolates on chromosome 5 (P5); six were isolate‐specific for isolate 09‐051 on chromosomes 10 (P10) and Pc, and two for isolate 07‐127 on chromosomes 11 (P11) and Pb. The QTLs in common with the major effect on the resistance for two isolates explained 20·0–48·2% of phenotypic variation. The isolate‐specific QTLs explained 6·0–17·4% of phenotypic variation. The result confirms a gene‐for‐gene relationship between C. annuum and P. capsici for root rot resistance.     

Comparative enzymatic hydrolysis of phytate in various animal feedstuff with two different phytases

Bacillus amyloliquefaciens DS11 phytase (DS11 phytase) and Aspergillus ficuum phytase (AF phytase) activities were investigated by measuring the release of phosphate from phytate in animal feedstuff such as wheat bran, corn meal, soybean meal and rice flour at pH 5 and 7. In all the tested feedstuff, the enzymatic activity of DS11 phytase was more active at pH 7, but that of AF phytase was more active at pH 5. From these results, the phytate in the gastrointestinal tract could be degraded in the small intestine or stomach by DS11 or AF phytase, respectively. In conclusion, the results presented in this paper indicated that different combination ratios of DS11 and AF phytase, depending on the kind of feedstuff, might effectively induce more enzymatic activity both in the stomach and small intestine in terms of the pH of the gastrointestinal tract.     

The presence of megamitochondria in the ellipsoid of photoreceptor inner segment of the zebrafish retina

Although the megamitochondria (MM) were localized in various pathological conditions, normal retina of some mammalian species was reported to include MM for various physiological roles. However, it was not clearly confirmed whether the MM is present in the retina of lower vertebrate as well. In this study, we tried to show the presence of the MM in the zebrafish retina using electron microscopic technique. In all the photoreceptors including rods, cones and double cones of the zebrafish retina, MM were observed in the ellipsoid of inner segment. In the photoreceptor epllipsoid of the zebrafish retina, the mitochondria located in the central portion of the ellipsoid had a highly electron-dense matrix, which were accompanied by the mitochondria with electron-lucent matrix in the apical portion of the ellipsoid. The presence of MM was more clearly discernable in the rods, which were localized under the double cones. This finding is somewhat different from those observed in the previous studies because MM were localized in the inner segment of cones, but were not in those of rods in the case of mammalian retina. Although the exact physiological meaning for the presence of MM in some vertebrate species should be further studied, the present study could show that the MM in the ellipsoid of the retinal photoreceptors was not only restricted in some mammalian species.     

Galanin-immunoreactive cells and their relation to calcitonin gene-related peptide-, substance P- and somatostatin-immunoreactive cells in rat lumbar dorsal root ganglia

We report upon the distribution of galanin-immunoreactive (GAL-IR) cells in the lumbar dorsal root ganglia (DRG) of the rat, and upon the distribution of GAL-IR cells, which also contain calcitonin gene-related peptide (CGRP)-, substance P (SP)- and somatostatin (SOM)-immunoreactivity. Neuropeptide-immunoreactive lumbar DRG cells were 55.8% for CGRP, 12.7% for SP, and 6.5% for GAL in lumbar DRG cells. There was no significant difference between the right and left DRGs (L1-L6) for any neuropeptide-immunoreactive cell (P < 0.01). In terms of size distribution, CGRP-immunoreactive cells were identified below 1500 microm2, and SP-, and GAL-IR cells below 600 microm2. Neuropeptide immunoreactive cells showed various immunoreactivities in the cytoplasm according to each neuropeptide. CGRP and SP immunoreactive cells were colocalized with GAL immunoreactive cells in the serial sections about 83.3 and 60% respectively, but SOM colocalizing with GAL-IR cells were not in evidence. The current results confirm and extend previous results, and show that neuropeptides can coexist in single sensory neurones of the rat DRG. In addition, our results demonstrate that the normal distribution of some neurotransmitters modulating sensory action in Wistar Kyoto rat, make this model more prone to develop neuropathic pain than Sprague-Dawley rat.     

Morphological characteristics of C1 and C2 adrenergic neurone groups in marmoset monkey brainstem by using antibody against phenylethanolamine-N-methyltransferase

This work describes a mapping study of phenylethanolamine-N-methyltransferase (PNMT) immunoreactive neurones and fibres in the medulla oblongata of the marmoset monkey, Callithrix jacchus. Two groups of PNMT-immunoreactive neurones were found in the marmoset monkey medulla oblongata: a ventrolateral (C1 group) and a dorsomedial PNMT-immunoreactive cells group (C2 group). The PNMT-immunoreactive cells in the ventrolateral group C1 were found to be located around the lateral reticular nucleus. The PNMT-immunoreactive somata within the ventrolateral medulla are round to oval, and mostly multipolar with branched processes. In the dorsomedial group C2, PNMT-immunoreactive cell bodies appeared near the obex. The majority of the dorsomedial PNMT-immunoreactive neurones were observed in the nucleus tractus solitarius; although some were present in the dorsal motor nucleus of the vagus. The PNMT-immunoreactive somata in the dorsomedial medulla were small and round or ovoid. These results provide information upon the adrenergic system in the medulla oblongata of a species that presents a useful model of a small primate brain, the marmoset monkey.     

The activity of avian Rhodanese

1. A survey of rhodanese activity (thiosulphate : cyanide sulphur transferase) in the tissues of the domestic fowl revealed that the highest activity occurred in the kidney, approximately twice that in the liver, 316 and 141 μmol SCN formed/min g protein, respectively.

2. In sparrows, pigeons and ducks, liver and kidney activities tended to be similar and increased with body size.

3. Relationships were found between enzyme activity and body weight to a fractional power, indicating relatively higher activity in smaller birds.     

Evaluation of Major Dietary Ingredients in Diverse Oats (Avena sativa L.) Germplasm

Journal of Crop Science and Biotechnology - Oat (Avena sativa L.) is one of the world’s most important cereal crops, owing to its use as an important source of essential nutrients for both...