Voluntary food intake variation in chickens on lysine-free diet is attributed to the plasma lysine concentration.

Growing chickens decrease their voluntary food intake when they receive a diet deficient in a single essential amino acid. Our previous studies suggest that the decreased food intake was associated with some metabolic changes. In order to reveal the involvement of plasma lysine fluctuations in the reduction of food intake, we examined whether maintaining the plasma lysine concentration in chickens on a lysine-free diet (the purified diet contained no lysine) restored the food intake to that of the control (lysine hydrochloride 11.9 g/kg) group. Male egg-type chickens at 21 d of age were injected with lysine at doses of 0.1 g/ml one hour after presenting the lysine-free diet. This injection increased the plasma lysine concentration one hour later and kept it similar to that of the control group for the following 2 h. Chickens ate the lysine-free diet as much as the control diet when their plasma lysine concentration was kept at a similar level to the control group. Injection of saline or alanine (0.12 g, isonitrogenous to lysine 0.1 g) into the crop of chickens on the lysine-free diet did not bring about the variations of food intake and plasma lysine concentrations as observed in those with lysine. 4. These findings show that the food intake variation was attributed to the plasma lysine concentration in the chickens on the lysine-free diet.

The use of warmed water treatment to induce protective immunity against the bacterial cold-water disease pathogen Flavobacterium psychrophilum in ayu (Plecoglossus altivelis).

We investigated the induction of protective immunity against bacterial cold-water disease (BCWD) caused by Flavobacterium psychrophilum by warmed water treatment in ayu (Plecoglossus altivelis). Fish were immersed in a live bacterial suspension (107 CFU mL⁻¹) for 30 min and placed in 700 L concrete tanks. The 28 °C warmed water treatment lasted 3 days and began 1, 6, and 24 h after immersion in the live bacterial suspension. A naïve control fish group was immersed in a sterilized modified Cytophaga (MCY) broth instead of the bacterial suspension. Fourteen days after the immersion, agglutination antibody titers against F. psychrophilum were measured by using micro-titer methods. Fish were then exposed to a bacterial bath to infect them with live F. psychrophilum, and cumulative mortality was monitored. Fish treated with warmed water at 1, 6, and 24 h after immersion in the live bacterial suspension had cumulative mortalities of 36%, 30%, and 18%, respectively, all of which were significantly lower than the cumulative mortality of the naïve control fish (90%). Treated fish also showed high antibody titers against F. psychrophilum in agglutination tests. These results demonstrate that warmed water treatment could not only cure BCWD but also immunize the fish against the causative agent F. psychrophilum.

A novel genotyping technique for distinguishing between Flavobacterium psychrophilum isolates virulent and avirulent to ayu, Plecoglossus altivelis altivelis (Temminck & Schlegel).

We developed a simple genotyping method for Flavobacterium psychrophilum for analysing two single nucleotide polymorphisms (SNPs) in the gyrA gene and to distinguish between isolates that are virulent and avirulent to ayu, Plecoglossus altivelis altivelis (Temminck & Schlegel). The genotyping method is an on/off switch assay and is based on the polymerase chain reaction technique with phosphorothioated primers. We classified 232 isolates from four families of fish (i.e. Plecoglossidae, Osmeridae, Cyprinidae and Salmonidae) into four genotypes (G-C, A-T, A-C and G-T). The G-C type isolates exhibited strong pathogenicity to ayu, whereas the A-T and G-T types did not show any pathogenicity to this species. The A-C type exhibited no or weak pathogenicity to ayu. These results indicate that genotyping F. psychrophilum isolates with two SNPs from gyrA can clearly distinguish between isolates potentially harmful to ayu (G-C type) and those that are potentially not harmful or less harmful (A-C, A-T and G-T type). The on/off switch assay provides a quick, simple, and very powerful DNA genotyping technique for F. psychrophilum isolates.

Oral administration of lysine restores food intake and ventromedial hypothalamic dopamine in chicken on a lysine-free diet.

1. This study was conducted to examine whether oral administration of lysine solution affect food intake and the ventromedial hypothalamic (VMH) monoamines in chickens fed on a lysine-free diet. 2. Chickens were assigned to four treatment groups. Two groups of chickens were given two different doses of lysine solution (0.1 g and 0.07 g in 1 ml of saline) exogenously (orally) while being fed on a lysine-free diet, and these results were compared with a control diet plus saline group. Another group of chickens was fed on a lysine-free diet without lysine supplementation, and their results were compared with the lysine treated groups. The extracellular dopamine (DA), norepinephrine (NE) and serotonin (5-HT) in the VMH of freely moving chicken were measured by in vivo microdialysis. 3. There was no significant difference in food intake between the control diet and the lysine supplemented groups during the time-course of the experiments. Food intake significantly decreased at 4, 5 and 6 h in the lysine-free diet plus saline group compared with the lysine supplemented groups. Of the VMH monoamines, the DA concentration remained close to the baseline in the lysine supplemented groups. This DA concentration was significantly lower than the baseline in the lysine-free diet plus saline group at 3.5 h and thereafter. 4. No significant difference from the baseline was observed for NE in the lysine-free diet plus saline group. The 5-HT concentrations were close to the baseline for all groups throughout the experiments. 5. The findings suggest that oral administration of lysine solution to chickens fed on a lysine-free diet restored food intake which was associated with the variations of VMH DA concentration.

The use of SPECT/CT to assess resorptive activity in mandibular condyles.

The aim of this study was to clarify the clinical significance of bone metabolism in the mandibular condyles in determining condylar resorptive changes. Twelve condyles of patients with idiopathic condylar resorption and degenerative joint disease were analysed using 99mTc HMDP SPECT/CT at baseline and subsequent computed tomography during the follow-up period. Twenty-two healthy condyles were enrolled as controls. After generating three-dimensional SPECT/CT images, two independent observers scored the degree of condylar uptake and measured the morphological changes in the condylar height and condylar volume. In the group with positive condylar uptake, the follow-up computed tomography showed significant decreases in condylar height (-1.69 ± 0.93 mm) and condylar volume (-12.51 ± 10.30%) when compared to healthy controls (condylar height, 0.09 ± 0.54 mm; condylar volume, -0.29 ± 4.22%) (P < 0.001). Moreover, the degree of uptake correlated with the changes in condylar height (observer 1, P = 0.012; observer 2, P = 0.039) and condylar volume (observer 1, P = 0.005; observer 2, P = 0.037). These results suggest that condylar bone metabolism is closely related to the resorptive activity. Thus, SPECT/CT would be useful in the prognostic evaluation or determination of treatment strategies for idiopathic condylar resorption and degenerative joint disease.

Optimization of iron oxide nanoparticle detection using ultrashort echo time pulse sequences: comparison of T1, T2*, and synergistic T1- T2* contrast mechanisms.

Iron oxide nanoparticles (IONPs) are used in various MRI applications as negative contrast agents. A major challenge is to distinguish regions of signal void due to IONPs from those due to low signal tissues or susceptibility artifacts. To overcome this limitation, several positive contrast strategies have been proposed. Relying on IONP T(1) shortening effects to generate positive contrast is a particularly appealing strategy because it should provide additional specificity when associated with the usual negative contrast from effective transverse relaxation time (T(2)*) effects. In this article, ultrashort echo time imaging is shown to be a powerful technique which can take full advantage of both contrast mechanisms. Methods of comparing T(1) and T(2)* contrast efficiency are described and general rules that allow optimizing IONP detection sensitivity are derived. Contrary to conventional wisdom, optimizing T(1) contrast is often a good strategy for imaging IONPs. Under certain conditions, subtraction of a later echo signal from the ultrashort echo time signal not only improves IONP specificity by providing long T(2)* background suppression but also increases detection sensitivity, as it enables a synergistic combination of usually antagonist T(1) and T(2)* contrasts. In vitro experiments support our theory, and a molecular imaging application is demonstrated using tumor-targeted IONPs in vivo.

Accuracy of Le Fort I osteotomy with combined computer-aided design/computer-aided manufacturing technology and mixed reality.

The aim of this study was to verify the reproducibility and accuracy of preoperative planning in maxilla repositioning surgery performed with the use of computer-aided design/manufacturing technologies and mixed reality surgical navigation, using new registration markers and the HoloLens headset. Eighteen patients with a mean age of 26.0 years were included. Postoperative evaluations were conducted by comparing the preoperative virtual operation three-dimensional image (Tv) with the 1-month postoperative computed tomography image (T1). The three-dimensional surface analysis errors ranged from 79.9% to 97.1%, with an average error of 90.3%. In the point-based analysis, the errors at each point on the XYZ axes were calculated for Tv and T1 in all cases. The median signed value deviation of all calculated points on the XYZ axes was -0.03mm (range -2.93mm to 3.93mm). The median absolute value deviation of all calculated points on the XYZ axes was 0.38mm (range 0mm to 3.93mm). There were no statistically significant differences between any of the points on any of the axes. These values indicate that the method used was able to reproduce the maxilla position with high accuracy.

Fas gene mutation in the progression of adult T cell leukemia.

Fas antigen (Apo-1/CD95) is an apoptosis-signaling cell surface receptor belonging to the tumor necrosis factor receptor superfamily. Adult T cell leukemia (ATL) cells express Fas antigen and show apoptosis after treatment with an anti-Fas monoclonal antibody. We established the ATL cell line KOB, which showed resistance to Fas-mediated apoptosis, and found that KOB expressed two forms of Fas mRNA, the normal form and a truncated form. The truncated transcript lacked 20 base pairs at exon 9, resulting in a frame shift and the generation of a premature stop codon at amino acid 239. The same mutation was detected in primary ascitic cells and peripheral blood cells. The mutation was not detected in lymph node cells, however, although all of the primary ATL cells were of the same clonal origin. A retroviral-mediated gene transfer of the truncated Fas to Jurkat cells rendered the cells resistant to Fas-mediated apoptosis, suggesting a dominant negative interference mechanism. These results indicate that an ATL subclone acquires a Fas mutation in the lymph nodes, enabling the subclone to escape from apoptosis mediated by the Fas/Fas ligand system and proliferate in the body. Mutation of the Fas gene may be one of the mechanisms underlying the progression of ATL.

Effect of surgery for cannula implantation into the hypothalamus on the behaviour and neural activity of broiler chicks.

1. The effects of a surgical operation, to implant a guide cannula in the chick hypothalamus for microdialysis, on behavioural responses and neural activity in broiler chicks are described. 2. General behavioural activities (feeding, preening, sitting, drinking, cage pecking and beak wiping), open field and locomotor activity tests were conducted to evaluate the effects related to surgery in the immediate 4 d following this procedure. Perfusion of Ringer solution with high K(+) after 4 d of guide cannula implantation was used to estimate the neural activity resulting from surgery through stimulation of monoamine release by in vivo brain dialysis. 3. The results of direct behavioral observations indicated that the stress provoked by surgical guide cannula insertion caused behavioural alterations that are particularly evident in the immediate days following this procedure. Open-field tests on day 4 after surgery showed that, compared to the intact control chickens, the treated chicks had a shorter latency to ambulate and defecate, with more vocalisation. Locomotor activity was less in the treated chicks than in the controls. 4. After 4 d of guide cannula implantation, the serotonin concentration started to increase 30 min after the onset of perfusing high-K(+) Ringer solution. It reached its highest value at one hour, suggesting that the 4 d after surgery is enough to alleviate some neurochemical dysfunction resulting from surgery. The results of behavioural observations, open-field and locomotor activity tests indicate that the surgical operation caused stress and fear in chicks which persisted up to 4 d.

Omani-type spondyloepiphyseal dysplasia with cardiac involvement caused by a missense mutation in CHST3.

We describe a family with progressive skeletal dysplasia and severe spinal involvement, short stature, premature arthrosis and joint contractures diagnosed as spondyloepiphyseal dysplasia Omani type. Mutation analysis in CHST3, the gene encoding for the chondroitin 6-O-sulfotransferase-1 (C6ST-1), revealed a homozygous missense mutation (T141M) in exon 3 in all three affected members of the family. Using recombinant C6ST-1, we showed that the identified missense mutation results in a reduction of C6ST-1 activity to 24-29% of the wild type protein. In addition to the previously noted skeletal features, affected members of this family also had cardiac involvement including mitral, tricuspid and/or aortic regurgitations and type E brachydactyly.

Novel oligosaccharide has suppressive activity against human leukemia cell proliferation.

Various oligosaccharides containing galactose(s) and one glucosamine (or N-acetylglucosamine) residues with beta1-4, alpha1-6 and beta1-6 glycosidic bond were synthesized; Galbeta1-4GlcNH(2), Galalpha1-6GlcNH(2), Galalpha1-6GlcNAc, Galbeta1-6GlcNH(2), Galbeta1-4Galbeta1-4GlcNH(2) and Galbeta1-4Galbeta1-4GlcNAc. Galalpha1-6GlcNH(2) (MelNH(2)) and glucosamine (GlcNH(2)) had a suppressive effect on the proliferation of K562 cells, but none of the other saccharides tested containing GlcNAc showed this effect. On the other hand, the proliferation of the human normal umbilical cord fibroblast was suppressed by none of the saccharides other than GlcNH(2). Adding Galalpha1-6GlcNH(2) or glucosamine to the culture of K562 cell, the cell number decreased strikingly after 72 h. Staining the remaining cells with Cellstain Hoechst 33258, chromatin aggregation was found in many cells, indicating the occurrence of cell death. Furthermore, all of the cells were stained with Galalpha1-6GlcNH-FITC (MelNH-FITC). Neither the control cells nor the cells incubated with glucosamine were stained. On the other hand, when GlcNH-FITC was also added to cell cultures, some of them incubated with Galalpha1-6GlcNH(2) were stained. The difference in the stainability of the K562 cells by Galalpha1-6GlcNH-FITC and GlcNH-FITC suggests that the intake of Galalpha1-6GlcNH(2) and the cell death induced by this saccharide is not same as those of glucosamine. The isolation of the Galalpha1-6GlcNH(2) binding protein was performed by affinity chromatography (melibiose-agarose) and LC-MS/MS, and we identified the human heterogeneous ribonucleoprotein (hnRNP) A1 (34.3 kDa) isoform protein (30.8 kDa). The hnRNP A1 protein was also detected from the eluate(s) of the MelNH-agarose column by the immunological method (anti-hnRNP-A1 and HRP-labeled anti-mouse IgG (gamma) antibodies).

Electrical currents flow out of domes formed by cultured epithelial cells.

Domes are localized areas of fluid accumulation between a cultured epithelial cell monolayer and the impermeable substratum on which the cells are cultured in vitro. Dome formation has been documented in a variety of epithelial cell lines that retain their transepithelial transport properties in vitro. However, it is not known whether domes are predominantly areas of specific active transport, or, alternatively, are predominantly areas of relative weak attachment to the culture surface. In the present study we adapted a vibrating microelectrode, which can detect small currents flowing in extracellular fluid, to determine if current was flowing into or out of domes and thereby to determine if domes were specialized areas of active transport. We used alveolar type II cells as the main epithelial cell type because they readily form domes in vitro and because they transport sodium from the apical to the basal surface. We found that electrical current flowed out of domes. The direction of the current was independent of the size of a dome, of the age of an individual dome, and of the number of days in primary culture for alveolar epithelial cells. This current was inhibited by amiloride and ouabain and was dependent on sodium in the medium. We made similar observations (outward current from domes which is blocked by amiloride and by sodium substitution) with domes formed by the Madin-Darby canine kidney cell line. The data support the hypothesis that sodium is transported across the entire monolayer and leaks back mainly through the domes. We conclude that domes in epithelial monolayers are not predominantly special sites of active transport but are more likely simply areas of weak attachment to the substratum.

Basic evaluation of a novel 4D target and human body phantom.

Stereotactic body radiation therapy (SBRT) is usually verified with a dynamic phantom or solid phantom, but there is a demand for phantoms that can accurately simulate tumor dynamics within an individual that would allow customized validation in every patient. We developed a new 4D dynamic target phantom (multi-cell 4D phantom) that allows simulation of tumor movement in patients. The basic quality and dynamic reproducibility of this new phantom was verified in this investigation. The newly developed multi-cell 4D phantom comprises four main components: soft tissue, bones, lungs, and tumor (target). The phantom structure was based on computed tomography (CT) data of a male. In this study, we investigated the basic performance of a multi-cell 4D phantom. All the CT numbers of the phantom were very close to those of human data. The geometric maximum amplitudes were 4.57 mm in the lateral direction, 4.59 mm in the ventrodorsal direction, and 3.68 mm in the cranio-caudal direction. Geometric errors were 0.84, 0.58, and 0.40 mm, respectively. Movements of the abdominal surface were stable for 60 s. Repeated measurements show no actual differences in target movements between multiple measurements and indicated high reproducibility (r > 0.97). End-to-end tests using Gafchromic film revealed a gamma pass rate of 98% or above (2 mm/3%). Although our phantom performed limited reproducibility in the movement of the patient tumor at present, a satisfactory level of precision was confirmed in general. This is a very promising device for use in the verification of radiation therapy for moving targets.

Autocrine and/or paracrine growth of aggressive ATLL cells caused by HGF and c-Met.

Adult T-cell leukemia/lymphoma (ATLL) is a neoplasia characterized by the massive invasion of various organs by tumor cells. Previously, we found that expression of the gene for c-Met, a receptor tyrosine kinase for hepatocyte growth factor (HGF), was specific to the acute type among 41 patients with ATLL by microarray. First in the present study, we analyzed the survival of the patients in relation to expression of c-Met and HGF in ATLL cells. Expression of the former but not the latter was associated with poor prognosis. Then, we analyzed the growth of ATLL cells caused by HGF and c-Met. c-Met was expressed in 0/7 chronic ATLLs, 12/14 acute ATLLs, 1/1 IL-2-independent ATLL cell line and 1/7 IL-2-dependent ATLL cell lines as assessed by flow cytometry. HGF induced the proliferation of primary cells from most acute cases examined as well as the c-Met-positive KK1 cell line in contrast to c-Met-negative cells. HGF induced autophosphorylation of c-Met in c-Met-positive cells from an acute case and KK1 cells. The plasma level of HGF was elevated in acute as compared to chronic cases. The levels of HGF and/or IL-6 which induces the production of HGF by stromal cells, were elevated in the supernatant of short-term cultured cells from certain patients with acute or chronic disease. Finally, infiltrated ATLL cells and adjacent stromal cells in liver were shown to be positive for c-Met/HGF and HGF, respectively, in acute cases. Autocrine and/or paracrine growth caused by HGF and c-Met was suggested in aggressive ATLL cells secreting HGF and/or IL-6, respectively.

Simple and low-cost tabletop NMR system for chemical-shift-resolution spectra measurements.

We have been working on developing a low-cost tabletop NMR system. We reported that a field homogeneity as high as 50 ppm was achieved with a simple NMR magnet by employing two facing ferrite magnets with iron disks in between (Chonlathep et al., 2017). In this paper, we report two improvements added to our previous system: (1) an FPGA based signal processing unit to improve the S/N ratio and (2) a simple shimming mechanism to improve the field homogeneity. We obtained as high as 1 ppm field homogeneity in the best case. The signals from hydrogen nuclear spins in a methyl and carboxy group in acetic acid were resolved in NMR spectra.

Recent advances in the study of the biosynthesis and functions of sulfated glycosaminoglycans.

Recent cDNA cloning of the glycosyltransferases involved in the synthesis of the sulfated glycosaminoglycan sidechains of proteoglycans has provided important clues to answering long-standing questions concerning the mechanisms of both chain polymerization and the biosynthetic sorting of glucosaminoglycans (heparin/heparan sulfate) and galactosaminoglycans (chondroitin/dermatan sulfate). These biosynthetic mechanisms are crucial to the expression and regulation of the biological functions of glycosaminoglycans in development and pathophysiology.

A simple and low-cost permanent magnet system for NMR.

We have developed a simple, easy to build, and low-cost magnet system for NMR, of which homogeneity is about 4×10-4 at 57mT, with a pair of two commercially available ferrite magnets. This homogeneity corresponds to about 90Hz spectral resolution at 2.45MHz of the hydrogen Larmor frequency. The material cost of this NMR magnet system is little more than $100. The components can be printed by a 3D printer.

Usefulness and capability of three-dimensional, full high-definition movies for surgical education.

BACKGROUND: Because of changing surgical procedures in the fields of oral and maxillofacial surgery, new methods for surgical education are needed and could include recent advances in digital technology. Many doctors have attempted to use digital technology as educational tools for surgical training, and movies have played an important role in these attempts. We have been using a 3D full high-definition (full-HD) camcorder to record movies of intra-oral surgeries. METHOD: The subjects were medical students and doctors receiving surgical training who did not have actual surgical experience (n = 67). Participants watched an 8-min, 2D movie of orthognathic surgery and subsequently watched the 3D version. After watching the 3D movie, participants were asked to complete a questionnaire. RESULT: A lot of participants (84%) felt a 3D movie excellent or good and answered that the advantages of a 3D movie were their appearance of solidity or realism. Almost all participants (99%) answered that 3D movies were quite useful or useful for medical practice. CONCLUSIONS: Three-dimensional full-HD movies have the potential to improve the quality of medical education and clinical practice in oral and maxillofacial surgery.

Erratum to: Usefulness and capability of three-dimensional, full high-definition movies for surgical education.

[This corrects the article DOI: 10.1186/s40902-017-0107-3.].