Characterization of canine filaggrin: gene structure and protein expression in dog skin

Background – Filaggrin (FLG) is a key protein for skin barrier formation and hydration of the stratum corneum. In humans, a strong association between FLG gene mutations and atopic dermatitis has been reported. Although similar pathogenesis and clinical manifestation have been argued in canine atopic dermatitis, our understanding of canine FLG is limited. Hypothesis/Objectives – The aim of this study was to determine the structure of the canine FLG gene and to raise anti‐dog FLG antibodies, which will be useful to detect FLG protein in dog skin. Methods – The structure of the canine FLG gene was determined by analysing the publicly available canine genome DNA sequence. Polyclonal anti‐dog FLG antibodies were raised based on the canine FLG sequence analysis and used for defining the FLG expression pattern in dog skin by western blotting and immunohistochemistry. Results – Genomic DNA sequence analysis revealed that canine FLG contained four units of repeated sequences corresponding to FLG monomer protein. Western blots probed with anti‐dog FLG monomer detected two bands at 59 and 54 kDa, which were estimated sizes. The results of immunohistochemistry showed that canine FLG was expressed in the stratum granulosum of the epidermis as a granular staining pattern in the cytoplasmic region. Conclusions and clinical importance – This study revealed the unique gene structure of canine FLG that results in production of FLG monomers larger than those of humans or mice. The anti‐dog FLG antibodies raised in this study identified FLG in dog skin. These antibodies will enable us to screen FLG‐deficient dogs with canine atopic dermatitis or ichthyosis.     

Suitable inflow rate and furrow length for simplified surge flow irrigation

Paddy and Water Environment - In the arid lands covering Uzbekistan, furrow irrigation with low application efficiency is still widely used due to a lack of funds and shortage of labor. To save...     

A simple,low‐cost technique for in situ measurement of leaf P concentration in field‐grown rice

Simple tools for diagnosing crop nutritional status are in great demand by agricultural extension staff, particularly in low‐fertility environments. We developed the first practical method for in situ diagnosis of phosphorus (P) nutrition in field‐grown cereal crops by using a handheld colorimeter. The concentration of P in extracts from fresh leaves of lowland rice grown under various P availabilities was closely correlated with that of oven‐dried leaves measured with the standard molybdenum‐blue method in the laboratory, over a range of values from 0.319 to 1.420 mg P g ^?1 DW (r = 0.885^**, n = 14). It takes only 3–5 min to estimate the P concentration of plant samples using the new technique. Although not as accurate as the laboratory method, the new method can easily detect P deficiency of rice in the field (and possibly other cereal crops) without requiring costly, off‐site equipment. Thus, extension staff and agronomists can easily adopt this technique.     

Angiotensin I-converting enzyme-inhibitory peptides obtained from chicken collagen hydrolysate

In this study, collagen extracted from chicken legs (which are the yellow keratin parts containing a nail) was hydrolyzed with various enzymes, and the angiotensin I-converting enzyme (ACE)-inhibitory activity of each hydrolysate was determined. The hydrolysate by treatment with an Aspergillus species-derived enzyme had the highest activity (IC 50 = 260 microg/mL). The fraction of this hydrolysate obtained by ultrafiltration with a molecular-weight cutoff of 3000 Da (low fraction) had a stronger activity (IC 50 = 130 microg/mL) than the fractionated one. This fraction was further fractionated by HPLC, and the peptides in the fraction with high ACE-inhibitory activity were identified. The amino acid sequences of the four peptides were identified using a protein sequencer. These peptides were synthesized to confirm their ACE-inhibitory activities; this showed that peptides with a Gly-Ala-Hyp-Gly-Leu-Hyp-Gly-Pro sequence had the highest activity (IC 50 = 29 microM). When the low fraction was administered to spontaneous hypertensive rats, a decrease in their blood pressure was observed after 2 h of administration, and a significant decrease in blood pressure (-50 mmHg) was observed after 6 h. Moreover, long-term administration studies indicated that the low fraction showed a significant suppression of increased blood pressure.     

Molecular characterization of an H1N2 swine influenza virus isolated in Miyazaki, Japan, in 2006

Swine influenza virus (SIV) was isolated from a farm in Miyazaki Prefecture in Japan in July 2006. An isolate was genetically subtyped as H1N2 and was designated A/swine/Miyazaki/1/2006 (H1N2). The nucleotide sequences of all eight viral RNA segments were determined, and then phylogenetic analysis was performed using the neighbor-joining method. All segments were shown to be closely related to those of Japanese SIV H1N2 isolates, which have been circulating since the 1980s. The results indicate the persistence of the SIV H1N2 subtype in the Japanese pig population for more than two decades and emphasize the importance of continuous surveillance for SIV.     

Histopathological characteristics of hepatic lipogranulomas with portosystemic shunt in dogs

In the canine liver with portosystemic shunts (PSS), focal lesions consisting of cells with cytoplasmic brown pigments and lipid vacuoles are often observed in the hepatic parenchyma. Termed lipogranulomas, their histopathological characteristics have been little studied. In the present study, we examined liver biopsy samples from 144 dogs (age: 3 months-16 years; 65 PSS and 79 non-PSS cases), and investigated the histopathological characteristics, incidence, and density of lipogranulomas. Lipogranulomas were detected histopathologically in 55.4% of PSS dogs. The lesions were then grouped into 3 types according to the amount of cytoplasmic lipid vacuoles and brown pigments. The pigments were positive for Berlin blue, PAS, and Sudan black B, but negative with the Hall method. The majority of the cells were immunohistochemically positive for macrophage scavenger receptor, class A (MSR-A), while no cells were positive for hepatocyte-antigen and albumin. The cytoplasmic pigments were recognized as electron-dense microgranular materials by electron microscopy. The incidence of lipogranulomas was significantly higher in the PSS group than non-PSS group when dogs less than 1 year old were excluded. The lipogranuloma density in the liver was significantly higher in the PSS group. It is concluded that lipogranulomas are frequently observed in liver biopsies of canine PSS especially in dogs more than 1 year old. The lesions consisted of Kupffer cells and/or macrophages, and the cytoplasmic brown pigments are ceroid and hemosiderin. The pathogenesis of lipogranuloma in PSS needs to be clarified.     

Storage of plug seedlings of tomato under limited fertilisation,and growth,flowering and yield after planting

We stored plug seedlings of tomato (Solanum lycopersicum L. cv. Momotaro) under limited fertilisation (LF) for 12 weeks, and evaluated growth and development during storage and after planting. Seedlings in the LF treatment were grown in substrate containing starter fertiliser and irrigated with water. Controls (CT) were irrigated with nutrient solution. Stem growth, leaf growth, and biomass accumulation slowed or ceased during storage of LF seedlings, while CT seedlings showed increases in these parameters. At 2 weeks after planting, the stem length of LF seedlings was shorter than that of CT seedlings, but the two seedling types had similar numbers of leaves. At harvest, LF and CT plants had similar numbers of leaves under the fruit trusses, but the trusses formed at lower heights in LF than in CT seedlings. LF and CT plants produced similar numbers of fruits and similar yields from the first to third trusses. These results indicate that tomato plug seedlings can be stored for more than 2 months under LF.     

Characterisation of starch properties and physical characteristics in buckwheat (Fagopyrum esculentum Moench.) mutant lacking accumulation of ‘granule-bound starch synthase a’

The concentration of amylose, which is synthesised using granule-bound starch synthase, affects the physical properties of food. However, no studies have focused on starch properties and physical characteristics of low-amylose buckwheat. Here, we hypothesised that low-amylose buckwheat would be useful to produce new buckwheat products because low-amylose characteristics change the texture of buckwheat food. In this study, we bred relatively low-amylose buckwheat compared to wild type and investigated the causative genes of the traits, starch properties and physical properties of noodles. In the GBSSa mutant, the amylose concentration was lower than that in the wild type. Compared with the wild type, the mutant exhibited the following traits: Amylose concentration decreased by approximately 2%, setback in the Rapid Visco Analyzer decreased by 30 points and the physical characteristics of noodles in the sensory analysis were soft and sticky. These results suggest that this trait may be useful for changing the texture of foods. In addition, the mutant is promising for producing new foods with physical characteristics that are different from those of the wild type.