Impact of amino acid 233 in Tax on bovine leukemia virus infection in Japanese Black cattle.

Bovine leukemia virus (BLV) is an economically important pathogen that both causes fatal enzootic bovine leukosis (EBL) and reduces lifetime milk production, reproductive efficiency, carcass weight, and longevity in dairy cows. The virus can be divided into two categories based on the amino acid at position 233 in Tax protein, which activates viral transcription and probably plays crucial roles in leukemogenesis. We recently reported that early-onset EBL in Japanese Black (JB) cattle was frequently caused by L233-Tax-carrying virus. This study examined the impact of BLV infection, the proviral load (PVL), and amino acid 233 in Tax on the outcomes of JB cattle. We measured PVL in cattle enrolled between February 2016 and December 2018, determined the Tax type of the isolates, and performed follow-up until March 2022. The results demonstrated that BLV infection increased the risk of involuntary culling and mortality in JB cattle in a PVL-dependent manner. Infection with L233-Tax-carrying virus increased the likelihood of mortality by 1.6-fold compared with the effects of P233-Tax-carrying virus infection. Intrauterine and perinatal infections were frequently caused by L233-Tax-carrying virus, and these infections were likely to influence the early onset of EBL in JB cattle. Conversely, breeding cows infected with P233-Tax-carrying virus were often eliminated by involuntary culling. These findings indicate that amino acid 233 in Tax has importance in terms of preventing economic loss attributable to EBL in JB cattle.

Characteristic endoscopic findings of gastrointestinal malignant lymphomas other than mucosa-associated lymphoid tissue lymphoma.

Background and study aims: The gastrointestinal (GI) tract is the most common site of extra-nodal involvement for non-Hodgkin's lymphoma (NHL). The features of GI NHLs remain unclear. The aim of this study was to clarify endoscopic characteristics of GI NHLs. Patients and methods: We retrospectively analyzed the morphological characteristics of 63 GI malignant lymphomas other than mucosa-associated lymphoid tissue lymphoma. Lesions were diagnosed between 2005 and 2020. Macroscopic findings were classified into five subtypes: superficial (S); protruding without ulcer (P); protruding with ulcer (PU); fungating (F); and multiple nodules (MN). Results: Thirty-one lesions in the stomach were classified as S type in 3 cases (9.6%), P type in 6 (19%), PU type in 13 (42%), and F type in 9 (29%). In the stomach, the ulcerated phenotype was more frequent for diffuse large B-cell lymphoma (DLBCL) (89.5%) than for other histological types (41.7%; P = 0.01). In the intestine, 23 tumors were classified as S type in 4 cases (17%), P type in 1 (4%), PU type in 6 (26%), F type in 1 (4%), and MN in 11 (48%). Eleven of the 14 cases (78.6%) of intestinal follicular lymphoma lesions showed MN type. In the colon, eight tumors were classified as S type in 2 cases (25%), P type in 2 (25%), PU type in 1 (13%), and F type in 3 (38%). Conclusion: We have clarified the endoscopic features of GI NHL using macroscopic classifications. The ulcerated phenotype was the most frequent endoscopic finding for DLBCL.

What are the risk factors for postoperative infections of third molar extraction surgery: A retrospective clinical study?

BACKGROUND: This study aimed to identify (1) the predilection site of postoperative infection after third molar extraction surgery, (2) risk factors associated with postoperative infection, and (3) the cause of the difference between delayed- and early-onset infections. MATERIALS AND METHODS: This retrospective study included 1010 patients (396 male, 614 female) who had ≥1 third molars extracted (2407; 812 maxilla, 1595 mandible). The risk factors were classified as attributes, general health, anatomic, and operative. Outcome variables were delayed- and early-onset infections. RESULTS: Postoperative infection was completely absent in the maxilla, and all infections occurred in the mandible, with a probability of 1.94% (31/1595). Bivariate analysis for postoperative infection showed depth of inclusion and intraoperative hemostatic treatment to be significantly associated with the development of infections. Bivariate analysis for delayed- and early-onset infections showed simultaneous extraction of the left and right mandibular third molars to be prominent risk factors. CONCLUSIONS: Postoperative infection occurs mainly in the mandible, and that in the maxilla is very rare. The risk of postoperative infection in the mandible was found to be related to the depth of inclusion and intraoperative hemostatic treatment. Simultaneous extraction of the left and right mandibular third molars appear to increase the risk of delayed-onset postoperative infection.

Author Correction: Self-organization of a human organizer by combined Wnt and Nodal signalling.

Ref. 7 from Benvenisty and colleagues was inadvertently omitted; this has now been cited in the text and added to the reference list, and subsequent references have been renumbered. The Letter has been corrected online.

Is the removal of osteosynthesis plates after orthognathic surgery necessary? Retrospective long-term follow-up study.

The removal of titanium miniplates is a controversial topic in oral and maxillofacial surgery. This retrospective study examined the timing of and reasons for titanium plate removal after orthognathic surgery. The study included 240 orthognathic surgery patients (71 male, 169 female; age range 16-55 years, mean 25.0±8.8 years) who had maxillofacial osteosynthesis plates inserted or inserted and then removed at the Division of Oral and Maxillofacial Surgery, Kagawa Prefectural Central Hospital, between April 2003 and March 2017. During the study period, a total of 717 miniplates were inserted in the 240 patients, and 71 of the patients (29.6%) had 236 plates (32.9%) removed. Ten patients (14.1%) had their plates removed within a year due to early complications. Although no patient had their plate removed due to complications at 1-5 years postoperative, a further 14 patients (19.7%) had their plates removed after more than 5 years of long-term follow-up due to plate-related complications. Complications requiring plate removal were evidently biphasic, occurring within 1 year after the operation and at ≥5 years after the operation. Therefore, after confirming postoperative bone healing, it is necessary to explain to patients the risks of plate removal and the importance of long-term follow-up.

Impact of prestorage leucoreduction of autologous whole blood on length of hospital stay with a subgroup analysis in bilateral hip arthroplasty.

BACKGROUND: Although prestorage leucoreduction (LR) of blood components for transfusion has gained favour around the world, evidence of its beneficial clinical effects is ambiguous. STUDY DESIGN AND METHODS: To reveal whether leucocytes and/or platelets in transfused blood are related to transfusion-related adverse effects, a prospective randomized crossover study was performed on patients who donated autologous blood prior to elective surgery. Among 1487 primary enrolees, a total of 192 patients undergoing two-stage, bilateral total hip arthroplasty were randomized to receive autologous blood that was either prestorage leucoreduced, or not, for the first procedure. For the second procedure, each patient was crossed over to receive alternatively processed autologous blood. Length of hospital stay served as a primary end-point, with perioperative infectious/thrombotic complications, pre- and postoperative laboratory values, and body temperature serving as secondary endpoints. RESULTS: No significant differences emerged between prestorage LR and non-LR cohorts in length of hospital stay, as well as perioperative infectious/thrombotic complications, postoperative body temperature and duration of fever. Postoperative laboratory values including white blood cell counts and C-reactive protein levels had no significant differences. CONCLUSION: This study could not prove any superiority of prestorage LR over non-LR for autologous whole blood among patients who underwent total hip arthroplasty.

Self-organization of a human organizer by combined Wnt and Nodal signalling.

In amniotes, the development of the primitive streak and its accompanying 'organizer' define the first stages of gastrulation. Although these structures have been characterized in detail in model organisms, the human primitive streak and organizer remain a mystery. When stimulated with BMP4, micropatterned colonies of human embryonic stem cells self-organize to generate early embryonic germ layers 1 . Here we show that, in the same type of colonies, Wnt signalling is sufficient to induce a primitive streak, and stimulation with Wnt and Activin is sufficient to induce an organizer, as characterized by embryo-like sharp boundary formation, markers of epithelial-to-mesenchymal transition and expression of the organizer-specific transcription factor GSC. Moreover, when grafted into chick embryos, human stem cell colonies treated with Wnt and Activin induce and contribute autonomously to a secondary axis while inducing a neural fate in the host. This fulfils the most stringent functional criteria for an organizer, and its discovery represents a milestone in human embryology.

Effect of low-temperature degradation on the mechanical and microstructural properties of tooth-colored 3Y-TZP ceramics.

The aim of the present study was to evaluate the effects of low-temperature degradation (LTD) induced by autoclaving on the mechanical and microstructural properties of tooth-colored 3 mol% yttria-stabilized tetragonal zirconia polycrystals (3Y-TZP). In total, 162 disc-shaped 3Y-TZP specimens were prepared. Two-thirds of the specimens were shaded by either the infiltration or powder mixing methods while the remaining specimens were used without coloring. The specimens were autoclaved at 134 °C for 0, 10, and 100 h to induce LTD (n=18 for each group). Chemical compositions were analyzed with X-ray fluorescence spectroscopy. Biaxial flexural strength was measured using a piston-on-three-ball test. The surface fraction and penetration depth of the monoclinic phase were examined using X-ray diffraction and scanning electron microscopy, respectively. The tooth-colored 3Y-TZP specimens contained Fe2O3 and Er2O3 (infiltration technique), and Fe2O3 (powder mixing method) at concentrations of<0.5 wt%. The tooth-colored 3Y-TZP had higher strength than the non-colored material after 100 h of autoclaving. In terms of surface fraction and penetration depth, the generation of monoclinic phase was significantly lower in the tooth-colored 3Y-TZP than in the non-colored material. The tooth-colored 3Y-TZP possessed equivalent biaxial flexural strength to that of the non-colored material and higher resistance to LTD regardless of the coloring technique (infiltration technique or powder mixing method) when the coloring pigments were contained at concentrations used in the present study.

Neurofilament Accumulation in Rabbit Retinas.

An eosinophilic body (EB) was observed in the inner nuclear layer and the outer plexiform layer of the anterior dorsal region of the retina in New Zealand White, Japanese White and Dutch rabbits. Immunohistochemistry confirmed that the EB was an accumulation of neurofilaments (NFs). Ultrastructurally, intermediate filaments of approximately 10 nm in diameter were observed in the EB, but there were no intracellular organelles. These results suggested that the NFs had accumulated in the neurites of the horizontal cells in the retina. This is the first description of a new pattern of NF accumulation in the mammalian retina. The prevalence of the EBs increased significantly in 44-56-week-old male Dutch rabbits (38.9 %) compared with 18-23-week-old (12.9 %) rabbits, suggesting that the formation of EBs in the rabbit retina could be an age-related change.

The influence of low-temperature degradation and cyclic loading on the fracture resistance of monolithic zirconia molar crowns.

The present study analyzed the kinetics of low-temperature degradation (LTD) in zirconia, and evaluated the influence of LTD and cyclic loading on the fracture resistance of monolithic zirconia molar crowns. Bar-shaped zirconia specimens were divided into nine groups and autoclaved at 134°C for 0-200h to induce LTD. The surface fraction and penetration depth of the monoclinic phase were examined using X-ray diffraction and scanning electron microscopy. Monolithic zirconia molar crowns were prepared for crown fracture testing. The crowns were autoclaved for 0-100h (n=6) and cemented to dies. Six crown-die samples that were not autoclaved and six samples that were autoclaved for 100h were subjected to cyclic loading with a load of 300N for 240,000 cycles. All samples were tested in a load-to-failure test. The monoclinic fraction on the surface increased with autoclaving time and reached a plateau after 50h. The depth of the monoclinic phase increased without reaching a plateau. The fracture load of the crowns significantly decreased from 5683N (SD: 342) to 3975N (SD: 194) after 100h of autoclaving. Cyclic loading did not significantly affect the fracture resistance of the crowns in all cases. Kinetic analysis showed no linear correlation between the surface fraction and depth of the monoclinic phase after 50h of autoclaving. Even though LTD increased the monoclinic phase, resulting in lower strength, the fracture resistance of the monolithic zirconia crowns was still sufficient to withstand the loading conditions in the molar regions.

Analysis of the essentiality of ROM2 genes in the pathogenic yeasts Candida glabrata and Candida albicans using temperature-sensitive mutants.

AIMS: To analyse the essentiality of the ROM2 genes originating from the pathogenic yeasts Candida glabrata and Candida albicans by using temperature-sensitive (ts) mutants. METHODS AND RESULTS: Based on the general concepts that ts mutations are generated by virtue of point mutation within essential genes, we have previously established a novel method (termed 'ETS system' for screening and identification of essential genes using ts mutants of C. glabrata). According to this ETS system, the present study successfully identified a putative C. glabrata ROM2 homologue as an essential gene that complements its point mutation (Cys-1275/Tyr substitution). The C. albicans ROM2 mutant (Cys-1281/Tyr), constructed patterned after this point mutation, also displayed ts phenotype. Both ts mutants recovered colony-forming ability, with concomitant suppression of lysis phenotype, at the elevated temperature in the presence of 1 mol l(-1) sorbitol as an osmotic stabilizer. Sequence alignment revealed that human genome possesses relatively low homology against Rom2 homologues, which are highly conserved among yeast species. CONCLUSIONS: ROM2 genes of C. glabrata and C. albicans are essential for viability, probably involved in cell wall integrity. SIGNIFICANCE AND IMPACT OF THE STUDY: ROM2 genes essential for both Candida species may be a potentially useful antifungal targets from chemotherapeutic viewpoint.

Effects of transforming growth factor beta 1 on the plasminogen activation system, collagen and integrin synthesis, and proliferation of rabbit mandibular condylar chondrocytes.

The objective of this study was to identify the mechanism by which mandibular condyle chondrocytes regulate the extracellular matrix. Primary rabbit condylar chondrocytes were isolated, cultured, and treated with transforming growth factor beta 1 (TGF-ß1). Cells were then assayed for the following: urokinase-type plasminogen activator (uPA) and its inhibitor (PAI-1), collagen types I and II, ß1 integrin expression, and proliferative activity. TGF-ß1 induced synthesis of collagen type II, αVß1 integrin, and PAI-1. TGF-ß1 induced the growth of chondrocytes and suppressed the synthesis of uPA. Chondrocyte regulation of the extracellular matrix is mediated by TGF-ß1. Synthesis of collagen type II, αVß1 integrin, and PAI-1 is induced, while uPA is suppressed. Also, TGF-ß1 induces cellular growth.

Mandibular fracture patterns consistent with posterior maxillary fractures involving the posterior maxillary sinus, pterygoid plate or both: CT characteristics.

OBJECTIVES: The aim of this study was to determine the incidence of posterior maxillary fractures involving the posterior maxillary sinus wall, pterygoid plate or both, unrelated to major midface fractures in patients with mandibular fractures, and to characterize associated fractures. METHODS: A CT study was performed in patients with mandibular fractures to identify posterior maxillary fractures. Patients aged under 16 years, those with mandibular fractures involving only dentoalveolar components and those with concurrent major midfacial fractures were excluded. RESULTS: 13 (6.7%) of 194 patients with mandibular fractures also had posterior maxillary fractures (case group). The injury pattern correlated with the external force directed to the lateral side of the mandible (p < 0.001), alcohol consumption (p = 0.049), the presence of multifocal fractures (p = 0.002) and the fracture regions in the symphysis/parasymphysis (p = 0.001) and the angle/ramus (p = 0.001). No significant difference between the case and non-case groups was seen for age, sex or cause of trauma. Non-displaced fractures in the ipsilateral posterior mandible occurred with significant frequency (p = 0.001) when the posterior maxillary fractures involved only the sinus. CONCLUSIONS: Mandibular fractures accompanied by posterior maxillary fractures are not rare. The finding of a unilateral posterior maxillary fracture on CT may aid the efficient radiological examination of the mandible based on possible patterns of associated fractures, as follows: in the ipsilateral posterior region as a direct fracture when the impact is a medially directed force, and in the symphysis/parasymphysis or contralateral condylar neck as an indirect fracture.

The retromandibular transparotid approach for reduction and rigid internal fixation using two locking miniplates in mandibular condylar neck fractures.

We evaluated the safety, efficacy, and morbidity associated with the treatment of displaced mandibular condylar neck fractures using a retromandibular transparotid approach to reduce and rigidly fix using two 2.0-mm locking miniplates. Our surgical inclusion criteria were: patient selection of open reduction and fixation, displaced unilateral condylar fractures with derangement of occlusion, and bilateral condylar fractures with an anterior open bite. The study group consisted of 19 patients who underwent surgery for 19 mandibular condylar neck fractures; patients were analyzed prospectively, with more than 6 months of follow-up, and were evaluated in terms of functional results, scar formation, postoperative complications, and stability of fixation. The results showed that functional occlusion identical to the preoperative condition and correct anatomical reduction of the condylar segments in centric occlusion, followed by immediate functional recovery, was achieved in all patients. No patient suffered from any major or permanent complication postoperatively, although there were two cases (11%) of temporary facial nerve palsy, which resolved completely within 3 months. Surgical scars were barely visible. The retromandibular transparotid approach with open reduction and rigid internal fixation for displaced condylar neck fractures of the mandible is a feasible and safe, minimally invasive surgical technique that provides reliable clinical results.

PI3 kinase directly phosphorylates Akt1/2 at Ser473/474 in the insulin signal transduction pathway.

Insulin stimulated translocation of the glucose transporter GLUT4 from the cytosol to the plasma membrane in a concentration (1  nM-1  µM)-dependent manner and increased glucose uptake in 3T3-L1 adipocytes. Insulin-induced GLUT4 translocation to the cell surface was prevented by the phosphoinositide 3 kinase (PI3K) inhibitor wortmannin, the 3-phosphoinositide-dependent protein kinase 1 (PDK1) inhibitor BX912 or the Akt1/2 inhibitor MK2206, and by knocking-down PI3K, PDK1 or Akt1/2. Insulin increased phosphorylation of Akt1/2 at Thr308/309 and Ser473/474, to activate Akt1/2, in the adipocytes. Insulin-induced phosphorylation of Akt1/2 was suppressed by wortmannin and knocking-down PI3K, while no significant inhibition of the phosphorylation was obtained with BX912 or knocking-down PDK1. In the cell-free Akt assay, PI3K phosphorylated Akt1 both at Thr308 and Ser473 and Akt2 at Ser474 alone. In contrast, PDK1 phosphorylates Akt1 at Thr308 and Akt2 at Thr309. The results of this study indicate that PI3K activates Akt1, independently of PDK1, and Akt2 by cooperating with PDK1 in the insulin signal transduction pathway linked to GLUT4 translocation.

Effect of ageing on healing of bilateral mandibular condyle fractures in a rat model.

To examine the hypothesis that conservative treatment is applicable to younger patients with bilateral mandibular condylar fractures, we studied the effect of ageing on the healing of bilateral mandibular condylar fractures in a rat model. Male Sprague-Dawley rats aged 3, 6, and 36 weeks (n=25/cohort, total n=75) were divided into a fracture group (n=12) and a sham control group (n=12); one rat from each cohort was used as a normal unoperated control. Cell proliferation was evaluated using the bromodeoxyuridine (BrdU) labelling index (LI). Osteochondrogenesis was assessed by the expression of Indian hedgehog (Ihh), type X collagen, and osteocalcin in the condylar head. Condylar fracture healing was found to be delayed by ageing. BrdU LI values in the fracture groups were higher in younger rats than in older rats at 8 weeks after fracture. The number of Ihh-positive cells in the fracture groups increased significantly up to 2 weeks after fracture, and then gradually decreased until 8 weeks after fracture. The findings of this study support the clinical concept of conservative treatment of bilateral condylar fractures in younger patients, but functional issues regarding ramus height and its consequences on occlusion have not been tested in this study.

Thyroid follicular adenoma with accumulation of collagen type IV in a common marmoset (Callithrix jacchus).

A thyroid tumour was identified in a 10-year-old male common marmoset (Callithrix jacchus). The tumour was encapsulated by fibrous connective tissue and compressed the adjacent normal thyroid. The tumour was composed of variably sized and irregularly shaped thyroid follicles lined by a single layer of columnar epithelial cells. Eosinophilic material at the base of the neoplastic cells stained black with periodic acid-methenamine silver and red with periodic acid-Schiff. Immunohistochemistry confirmed that this eosinophilic material was collagen type IV. Ultrastructurally, highly dense and amorphous material was observed at the base of the neoplastic cells. Small vesicles in the basolateral cytoplasm of the neoplastic cells contained similar material to that at the base of the cells. The tumour was diagnosed as a thyroid follicular adenoma with accumulation of collagen type IV. This is the first description of an endocrine tumour with accumulation of collagen type IV in animals.

A detailed description of an economical setup for electroporation of chick embryos in ovo.

One of the challenges of the postgenomic era is characterizing the function and regulation of specific genes. For various reasons, the early chick embryo can easily be adopted as an in vivo assay of gene function and regulation. The embryos are robust, accessible, easily manipulated, and maintained in the laboratory. Genomic resources centered on vertebrate organisms increase daily. As a consequence of optimization of gene transfer protocols by electroporation, the chick embryo will probably become increasingly popular for reverse genetic analysis. The challenge of establishing chick embryonic electroporation might seem insurmountable to those who are unfamiliar with experimental embryological methods. To minimize the cost, time, and effort required to establish a chick electroporation assay method, we describe and illustrate in great detail the procedures involved in building a low-cost electroporation setup and the basic steps of electroporation.

A detailed description of an economical setup for electroporation of chick embryos in ovo

One of the challenges of the postgenomic era is characterizing the function and regulation of specific genes. For various reasons, the early chick embryo can easily be adopted as an in vivo assay of gene function and regulation. The embryos are robust, accessible, easily manipulated, and maintained in the laboratory. Genomic resources centered on vertebrate organisms increase daily. As a consequence of optimization of gene transfer protocols by electroporation, the chick embryo will probably become increasingly popular for reverse genetic analysis. The challenge of establishing chick embryonic electroporation might seem insurmountable to those who are unfamiliar with experimental embryological methods. To minimize the cost, time, and effort required to establish a chick electroporation assay method, we describe and illustrate in great detail the procedures involved in building a low-cost electroporation setup and the basic steps of electroporation.

Barriers and motivators to blood donation among university students in Japan: development of a measurement tool.

BACKGROUND AND OBJECTIVES: Despite growing demand for transfusion, the number of voluntary young blood donors has steadily decreased over recent years in Japan. This study aimed to develop an easy-to-use survey tool to assess barriers and motivators to blood donation among Japanese university students. MATERIALS AND METHODS: We conducted cross-sectional studies at two universities in Fukushima Prefecture, Japan, in December 2011 (Stage 1) and February 2012 (Stage 2) using self-administered questionnaires. A short list of motivators and barriers to blood donation was developed from the open-ended questions asked of 50 students in Stage 1. In the Stage 2, we asked 105 students how important these items were when they decided whether or not to donate blood. Items showing a significant difference between donors and non-donors were kept in the final list. RESULTS: Overall, 56% of the 100 participants analysed in Stage 2 were men, and ages ranged from 19 to 24 with a median of 20 years. Comparison of motivators and barriers between donors and non-donors revealed that only barrier item 8 ('Frightened by blood donation') showed a significant difference (P = 0·0006) in an expected direction and with a consistency between two universities. CONCLUSIONS: This study identified fear as being the most significant barrier to blood donation among Japanese university students, which could be used as a single convenient indicator to assess their readiness to donate. More academic and clinical efforts are needed to understand and address students' fear towards blood donation in order to increase the donor pool in Japan.