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.

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.

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.

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.

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.].

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.

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.

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.

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.

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.

Transtumoral targeting enabled by a novel neuropilin-binding peptide.

We have recently described a class of peptides that improve drug delivery by increasing penetration of drugs into solid tumors. These peptides contain a C-terminal C-end Rule (CendR) sequence motif (R/K)XX(R/K), which is responsible for cell internalization and tissue-penetration activity. Tumor-specific CendR peptides contain both a tumor-homing motif and a cryptic CendR motif that is proteolytically unmasked in tumor tissue. A previously described cyclic tumor-homing peptide, LyP-1 (sequence: CGNKRTRGC), contains a CendR element and is capable of tissue penetration. We use here the truncated form of LyP-1, in which the CendR motif is exposed (CGNKRTR; tLyP-1), and show that both LyP-1 and tLyP-1 internalize into cells through the neuropilin-1-dependent CendR internalization pathway. Moreover, we show that neuropilin-2 also binds tLyP-1 and that this binding equally activates the CendR pathway. Fluorescein-labeled tLyP-1 peptide and tLyP-1-conjugated nanoparticles show robust and selective homing to tumors, penetrating from the blood vessels into the tumor parenchyma. The truncated peptide is more potent in this regard than the parent peptide LyP-1. tLyP-1 furthermore improves extravasation of a co-injected nanoparticle into the tumor tissue. These properties make tLyP-1 a promising tool for targeted delivery of therapeutic and diagnostic agents to breast cancers and perhaps other types of tumors.

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.

Evaluation of a simple loop-mediated isothermal amplification test kit for the diagnosis of tuberculosis.

OBJECTIVE: A new loop-mediated isothermal amplification (LAMP) test kit, including a simple DNA extraction device for the detection of Mycobacterium tuberculosis complex, was developed for commercial use and evaluated for its usefulness in diagnosing tuberculosis (TB). DESIGN: The LAMP test was performed using untreated and N-acetyl-L-cysteine (NALC) NaOH-treated sputum specimen. The efficiency of the kit was compared with other conventional laboratory examinations, including other nucleic acid amplification (NAA) tests. RESULTS: The sensitivity of LAMP using raw sputum (direct LAMP) in smear- and culture-positive specimens was 98.2% (95%CI 94.9-99.4), while the sensitivity in smear-negative, culture-positive specimens was 55.6% (95%CI 43.4-68.0). The diagnostic sensitivity of direct LAMP for the diagnosis of individuals with TB was 88.2% (95%CI 81.4-92.7). The sensitivity values of direct LAMP were slightly, but not statistically significantly lower than those of Cobas Amplicor MTB and TRC Rapid MTB, while the sensitivity of the LAMP test using NALC-NaOH treated sputum was significantly lower than other NAA tests (P < 0.05) for smear-negative, culture-positive specimens. The new commercial version of the LAMP kit was easy to handle and yielded results within 1 h of receiving sputum specimens. CONCLUSIONS: This test is considered a promising diagnostic tool for TB, even for peripheral laboratories with limited equipment, such as those in developing countries.

Potent suppression of stretch reflex activity after systemic or spinal delivery of tizanidine in rats with spinal ischemia-induced chronic spastic paraplegia.

BACKGROUND: Spasticity and rigidity are serious complications associated with spinal traumatic or ischemic injury. Clinical studies show that tizanidine (Tiz) is an effective antispasticity agent; however, the mechanism of this effect is still not clear. Tiz binds not only to α2-adrenoreceptors (AR) but also to imidazoline (I) receptors. Both receptor systems (AR+I) are present in the spinal cord interneurons and α-motoneurons. The aim of the present study was to evaluate the therapeutic potency of systematically or spinally (intrathecally [IT]) delivered Tiz on stretch reflex activity (SRA) in animals with ischemic spasticity, and to delineate supraspinal or spinal sites of Tiz action. EXPERIMENTAL PROCEDURES: Animals were exposed to 10 min of spinal ischemia to induce an increase in SRA. Increase in SRA was identified by simultaneous increase in recorded electromyography (EMG) activity and ankle resistance measured during computer-controlled ankle dorsiflexion (40°/3 s) in fully awake animals. Animals with increased SRA were divided into several experimental subgroups and treated as follows: (i) Tiz administered systemically at the dose of 1 mg kg(-1), or IT at 10 µg or 50 µg delivered as a single dose; (ii) treatment with systemic Tiz was followed by the systemic injection of vehicle, or by nonselective AR antagonist without affinity for I receptors; yohimbine (Yoh), α2A AR antagonist; BRL44408 (BRL), α2B AR antagonist; ARC239 (ARC), nonselective AR and I(1) receptor antagonist; efaroxan (Efa), or nonselective AR and I(2) receptor antagonist; idazoxan (Ida); (iii) treatment with IT Tiz was followed by the IT injection of selective α2A AR antagonist; atipamezole (Ati). In a separate group of spastic animals the effect of systemic Tiz treatment (1 mg/kg) or isoflurane anesthesia on H-reflex activity was also studied. RESULTS: Systemic and/or IT treatment with Tiz significantly suppressed SRA. This Tiz-mediated anti-SRA effect was reversed by BRL (5 mg kg(-1)), Efa (1 mg kg(-1)), and Ida (1 mg kg(-1)). No reversal was seen after Yoh (3 mg kg(-1)) or ARC (5 mg kg(-1)) treatment. Anti-SRA induced by IT Tiz (50 µg) was reversed by IT injection of Ati (50 µg). Significant suppression of H-reflex was measured after systemic Tiz treatment (1 mg/kg) or isoflurane (2%) anesthesia, respectively. Immunofluorescence staining of spinal cord sections taken from animals with spasticity showed upregulation of α2A receptor in activated astrocytes. CONCLUSIONS: These data suggest that α2A AR and I receptors, but not α2B AR, primarily mediate the Tiz-induced antispasticity effect. This effect involves spinal and potentially supraspinal sites and likely targets α2A receptor present on spinal neurons, primary afferents, and activated astrocytes. Further studies using highly selective antagonists are needed to elucidate the involvement of specific subtypes of the AR and I receptors in the antispasticity effect seen after Tiz treatment.

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.

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.

Activation of p53 by Nutlin-3a, an antagonist of MDM2, induces apoptosis and cellular senescence in adult T-cell leukemia cells.

It has been reported that the induction of cellular senescence through p53 activation is an effective strategy in tumor regression. Unfortunately, however, tumors including adult T-cell leukemia/lymphoma (ATL) have disadvantages such as p53 mutations and a lack of p16(INK4a) and/or p14(ARF). In this study we characterized Nutlin-3a-induced cell death in 16 leukemia/lymphoma cell lines. Eight cell lines, including six ATL-related cell lines, had wild-type p53 and Nutlin-3a-activated p53, and the cell lines underwent apoptosis or cell-cycle arrest, whereas eight cell lines with mutated p53 were resistant. Interestingly, senescence-associated-beta-galactosidase (SA-beta-gal) staining revealed that only ATL-related cell lines with wild-type p53 showed cellular senescence, although they lack both p16(INK4a) and p14(ARF). These results indicate that cellular senescence is an important event in p53-dependent cell death in ATL cells and is inducible without p16(INK4a) and p14(ARF). Furthermore, knockdown of Tp53-induced glycolysis and apoptosis regulator (TIGAR), a novel target gene of p53, by small interfering RNA(siRNA) indicated its important role in the induction of cellular senescence. As many patients with ATL carry wild-type p53, our study suggests that p53 activation by Nutlin-3a is a promising strategy in ATL. We also found synergism with a combination of Nutlin-3a and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), suggesting the application of Nutlin-3a-based therapy to be broader than expected.

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).