Rocuronium priming for tracheal intubation in COVID-19 patients.

Priming doses of non-depolarising neuromuscular blocking drugs given before administration of anaesthetic agents have been used to hasten the onset of neuromuscular blockade. In the settings of coronavirus disease 2019 (COVID-19), this could be used to reduce the apnoeic, and potentially aerosol-generating, window. To our knowledge, we report the first cases of tracheal intubation with rocuronium for COVID-19 using the priming principle. Both patients needed their tracheas intubated for severe hypoxia using a rapid sequence induction technique with a priming dose of rocuronium. Despite adequate pre-oxygenation a sudden, unexpected fall in arterial oxygen saturations was observed in both patients after administration of a priming dose of 2 mg of rocuronium. Clinicians should consider this possible risk associated with priming doses of neuromuscular blocking drugs in the management of patients with respiratory failure due to COVID-19.

Denture wearing during sleep doubles the risk of pneumonia in the very elderly.

Poor oral health and hygiene are increasingly recognized as major risk factors for pneumonia among the elderly. To identify modifiable oral health-related risk factors, we prospectively investigated associations between a constellation of oral health behaviors and incident pneumonia in the community-living very elderly (i.e., 85 years of age or older). At baseline, 524 randomly selected seniors (228 men and 296 women; mean age, 87.8 years) were examined for oral health status and oral hygiene behaviors as well as medical assessment, including blood chemistry analysis, and followed up annually until first hospitalization for or death from pneumonia. During a 3-year follow-up period, 48 events associated with pneumonia (20 deaths and 28 acute hospitalizations) were identified. Among 453 denture wearers, 186 (40.8%) who wore their dentures during sleep were at higher risk for pneumonia than those who removed their dentures at night (log rank P = 0.021). In a multivariate Cox model, both perceived swallowing difficulties and overnight denture wearing were independently associated with an approximately 2.3-fold higher risk of the incidence of pneumonia (for perceived swallowing difficulties, hazard ratio [HR], 2.31; and 95% confidence interval [CI], 1.11-4.82; and for denture wearing during sleep, HR, 2.38; and 95% CI, 1.25-4.56), which was comparable with the HR attributable to cognitive impairment (HR, 2.15; 95% CI, 1.06-4.34), history of stroke (HR, 2.46; 95% CI, 1.13-5.35), and respiratory disease (HR, 2.25; 95% CI, 1.20-4.23). In addition, those who wore dentures during sleep were more likely to have tongue and denture plaque, gum inflammation, positive culture for Candida albicans, and higher levels of circulating interleukin-6 as compared with their counterparts. This study provided empirical evidence that denture wearing during sleep is associated not only with oral inflammatory and microbial burden but also with incident pneumonia, suggesting potential implications of oral hygiene programs for pneumonia prevention in the community.

Distinct functional regions of the human polymeric immunoglobulin receptor.

The polymeric immunoglobulin receptor (pIgR) is a type I transmembrane protein that is expressed on the surfaces of glandular and intestinal epithelial cells. The extracellular portion of the pIgR is composed of six different domains. Domain 6 is involved in the enzymatic cleavage and release of the pIgR into the intestinal lumen as a free secretory component (fSC). A highly conserved 9-amino acid sequence is present in this region in various species. Although mutations in domain 6 are associated with particular diseases, such as IgA nephropathy and Epstein-Barr virus-related nasopharyngeal cancer, and the glutamic acid residues in the conserved 9-amino acid sequence are expected to be indispensable for the secretion of fSC, the importance of these residues has not been examined. In the present study, we attempted to examine the role of these residues in the enzymatic cleavage of the pIgR. The enzymatic cleavage of the pIgR was not affected by the presence of an alanine to valine substitution at position 580 or glutamine to alanine substitutions at positions 606 and/or 607, or the deletion of the whole 9-amino acid conserved sequence. Intriguingly, the 10 amino acid sequences flanking the N- and C-terminal ends of the conserved 9-amino acid sequence had opposite effects on pIgR cleavage. Namely, the N-terminal and C-terminal sequences enhanced and reduced pIgR cleavage efficiency, respectively. These results indicated that the pIgR can be divided into several functionally distinct regions.

The use of arthrography in the diagnosis of temporomandibular joint synovial chondromatosis.

Synovial chondromatosis (SC) involving the temporomandibular joint (TMJ) is very rare and can occur in either or both cavities. Differentiation of the affected cavity in SC is therefore as important as making the diagnosis. This report presents a case of SC in which both cavities were thought to be affected, but arthrography using cone beam CT (CBCT) allowed us to see that involvement was limited to the superior joint cavity. In addition, we describe the usefulness of arthrographic CBCT for diagnosis and treatment planning in SC of the TMJ.

Phase I/II trial of a biweekly combination of S-1 plus docetaxel in patients with previously treated non-small cell lung cancer (KRSG-0601).

BACKGROUND: Combination of S-1, an oral fluorouracil derivative, plus docetaxel against non-small cell lung cancer (NSCLC) showed promising efficacy but clinically problematic emesis. A phase I/II study utilising a new schedule for this combination was conducted. METHODS: A biweekly regimen of docetaxel on day 1 with oral S-1 on days 1-7 was administered to previously treated NSCLC patients. Doses of docetaxel/S-1 were escalated to 30/80, 35/80, and 40/80 mg m(-2), respectively, and its efficacy was investigated at the recommended dose below maximum tolerated dose (MTD). RESULTS: In phase I study employing 13 patients, dose-limiting toxicities were febrile neutropenia and treatment delay, with the respective MTDs for docetaxel 40 mg m(-2)/S-1 80 mg m(-2). In the phase II study, 34 patients were treated with docetaxel 35 mg m(-2)/S-1 80 mg m(-2) for a median cycle of 6. The response and disease control rates were 34.3% (95% confidence interval (CI), 18.6-50.0%) and 62.9% (95% CI, 46.8-72.9%), respectively. Median progression-free survival was 150.5 days. Haematologic grade 4 toxicities were observed in neutropenia (11.8%) and thrombocytopenia (2.9%). Regarding non-haematologic toxicities, including emesis, there were no grade 3/4 side effects. CONCLUSION: Combination of 1-week administration of S-1 with biweekly docetaxel is safe and active for NSCLC.

Nuclear factor kappa B plays a pivotal role in polyinosinic-polycytidylic acid-induced expression of human ß-defensin 2 in intestinal epithelial cells.

Intestinal epithelial cells (IECs) play an important role in protecting the intestinal surface from invading pathogens by producing effector molecules. IECs are one of the major sources of human beta-defensin 2 (hBD-2), and can produce it in response to a variety of stimuli. Although IECs express Toll-like receptor 3 (TLR-3) and can respond to its ligand, double-stranded RNA (dsRNA), hBD-2 expression in response to dsRNA has not been elucidated. In the present study, using an artificial analogue of dsRNA, polyinosinic-polycytidylic acid (poly I:C), we investigated whether the human IEC line, HT-29, can produce hBD-2 in response to poly I:C. HT-29 cells can express hBD-2 mRNA only when stimulated with poly I:C. The induction of hBD-2 mRNA expression was observed at 3 h after stimulation and peaked at 12 h of post-stimulation. Pre-incubation of the cells with nuclear factor kappa B (NF-κB)-specific inhibitor, l-1-4'-tosylamino-phenylethyl-chloromethyl ketone (TPCK) and isohelenine abolished the expression of hBD-2. Detection of the poly I:C signal by TLR-3 on the surface of HT-29 cells was revealed by pre-incubating the cells with anti-TLR-3 antibody. The 5'-regulatory region of the hBD-2 gene contains two NF-κB binding sites. A luciferase assay revealed the importance of the proximal NF-κB binding site for poly I:C-induced expression of hBD-2. Among NF-κB subunits, p65 and p50 were activated by poly I:C stimulation and accumulated in the nucleus. Activation of the p65 subunit was investigated further by determining its phosphorylation status, which revealed that poly I:C stimulation resulted in prolonged phosphorylation of p65. These results indicate clearly that NF-κB plays an indispensable role in poly I:C induced hBD-2 expression in HT-29 cells.

TGF-ß signaling in gingival fibroblast-epithelial interaction.

The underlying mechanism and the therapeutic regimen for the transition of reversible gingivitis to irreversible periodontitis are unclear. Since transforming growth factor (TGF)-ß has been implicated in differentially regulated gene expression in gingival fibroblasts, we hypothesized that TGF-ß signaling is activated in periodontitis-affected gingiva, along with enhanced collagen degradation, that is reversed by TGF-ß inhibition. A novel three-dimensional (3D) gel-culture system consisting of primary human gingival fibroblasts (GF) and gingival epithelial (GE) cells in collagen gels was applied. GF populations from patients with severe periodontitis degraded collagen gels, which was reduced by TGF-ß-receptor kinase inhibition. Up-regulation of TGF-ß-responsive genes was evident in GF/GE co-cultures. Furthermore, the TGF-ß downstream transducer Smad3C was highly phosphorylated in periodontitis-affected gingiva and 3D cultures. These results imply that TGF-ß signaling is involved in fibroblast-epithelial cell interaction in periodontitis, and suggest that the 3D culture system is a useful in vitro model for therapeutic drug screening for periodontitis.

Unusual radiographic appearance of ossifying fibroma in the left mandibular angle.

Ossifying fibroma is usually a unilocular lesion with a well-defined, thinly corticated margin radiographically, although various patterns have been noted. The patient was a 27-year-old woman with a painless radiolucent lesion demonstrated on panoramic radiography to involve the root-apex area of the left lower second and third molars. Radiographically, the lesion had some features of a benign tumour, such as an odontogenic myxoma. However, the deep invaginations towards the interalveolar septa suggested a simple bone cyst, whereas the irregular margin and lack of expansion or mandibular canal displacement were consistent with a malignant lesion. A hard tissue component was confirmed only by soft-tissue mode CT. Although this lesion was histopathologically diagnosed as ossifying fibroma, the conflicting imaging findings were challenging and very intriguing.

Three-dimensional reconstruction of a fibro-osseous lesion using binary images transformed from histopathological images.

The aim of this report was to introduce a new method of three-dimensional (3D) reconstruction for fibro-osseous lesions (FOLs) using binary images transformed from histopathological images and to describe its usefulness. A sample of multiconfluent FOL was used (one of the five types of FOL according to a radiographic classification) which was diagnosed histopathologically as ossifying fibroma. Approximately 30 pathological images were assembled into a composite image of the slide using Tiling Boutique software version 3 for Windows (Sanyo Electric, Osaka, Japan). The tiling images were transformed into 8-bit scale images and then into binary images using ImageJ software ver.1.37 (National Institutes of Health, Bethesda, MD). These images were used for 3D reconstruction using ImageJ software. Images were loaded at the same matrix size and were reconstructed into layers of two-dimensional image stacks, adjusted so that contiguous images were aligned based on their centre points, and arranged with long axes horizontal. 3D findings aided the visual understanding of morphological features in the lesion. The 3D reconstruction can be displayed with arbitrary rotation. In this case, the 3D reconstruction, using Real Image software version 4.01 for Windows (KGT, Tokyo, Japan), was created from an arbitrary section. This allowed us to determine the pattern of calcification between groups of connected osteoids and to compare the internal structure of such lesions that are not visible on histopathological findings. Differentiation of features was even more pronounced with a two colour display indicating fibrous connective tissue and osteoid tissue. A 3D reconstruction of a multiconfluent ossifying fibroma was created using binary images transformed from histopathological images. The quality of the images depends above all on the functionality of the image-processing software. Comparison of each pattern of FOL might allow more simple assessment of the morphological features of FOLs.

Central odontogenic fibroma with osteoid formation showing atypical radiographic appearance.

We recently encountered an interesting tumour, containing both a diffuse sclerotic border and calcified bodies, which was eventually diagnosed as a central odontogenic fibroma. The patient was a 40-year-old man with a painless radiolucent area in the crown area of the unerupted left lower third molar shown by panoramic radiography. Clinically, the lesion was thought to represent an odontogenic tumour involving a calcified body, i.e. calcifying cystic odontogenic tumour, calcifying epithelial odontogenic tumour or ossifying fibroma. Diagnosis by radiographic findings was extremely difficult.

Sero-epidemiology of sporadic acute hepatitis in Bangladesh: high prevalences of infection with type-B, type-E and multiple types of hepatitis virus.

In a recent investigation of hepatitis in Bangladesh, the sera from 74 adult patients (aged 15-67 years) who had been clinically diagnosed as cases of sporadic acute hepatitis were collected at various hospitals in and around Dhaka. Five cases were positive for IgM antibody against the hepatitis A virus and 30 were positive both for the surface antigen of the hepatitis B virus (HBV) and for IgM antibody against the HBV core (HBc). The six cases found positive for antibodies against the hepatitis D virus were all also positive for the HBV surface antigen but negative for anti-HBc IgM. Thirteen patients harboured hepatitis C virus RNA and 29 were positive for IgM antibody against the hepatitis E virus (HEV). There were 14 non-A-to-E subjects, whose illness was of unknown aetiology. Of the 83 infections with hepatitis viruses detected in the other 60 patients, 6%, 36%, 16%, 7% and 35% were of types A, B, C, D and E, respectively. Each of 28 of the patients (47% of those confirmed to have viral hepatitis) had concomitant infection with more than one type of hepatitis virus. The predominance of HBV and HEV infections and the high prevalence of multiple infection seen among these Bangladeshi cases have not been observed among hepatitis cases in developed countries.

Poly I:C-induced expression of intercellular adhesion molecule-1 in intestinal epithelial cells.

Intercellular adhesion molecul-1 (ICAM-1) is a transmembrane glycoprotein belonging to the immunoglobulin superfamily of adhesion molecules and plays perdominant roles in recruitment and trafficking of leucocytes to sites of inflammation. ICAM-1 expression in intestinal epithelial cells (IECs) is enhanced by several stimuli, such as proinflammatory cytokines, bacterial infections or pathogen-associated molecular patterns. One of these stimuli, double-stranded RNA (dsRNA), is a by-product of viral replication and can be recognized by its cognate receptor Toll-like receptor 3 (TLR-3). In spite of expression of both TLR-3 and ICAM-1 in IECs, correlation between TLR-3-signalling and ICAM-1 expression has never been examined in IECs. In the present study, we investigated whether poly I:C, an analogue of dsRNA, can stimulate the expression of ICAM-1 in IEC line, HT-29. Poly I:C-stimulation up-regulated the expression of ICAM-1 mRNA by real-time polymerase chain reaction. Enhanced expression of ICAM-1 was confirmed in protein level by immunofluoresense cell staining and enzyme-linked immunosorbent assay by measuring the released soluble ICAM-1 in culture supernatant. As the stimulation effect was reduced by pre-treatment of the cells with anti-TLR-3 antibody, poly I:C-binding signal was thought to be sensed by TLR-3 on the surface of HT-29. The results of luciferase assay and nuclear factor kappa-b (NF-kappaB) inhibitor treatment experiments indicated that the downstream signal was mainly transduced by transcription factor, NF-kappaB. All these results demonstrated the connection between TLR-3 signalling and ICAM-1 expression in HT-29 cells and indicated the importance of coordinated function of both innate and adaptive immunity against viral infections.

Correlation between histopathological image and radiographic image pattern in fibro-osseous lesions in relation to bone complexity and distribution.

OBJECTIVES: This study examined correlations between radiographic patterns and the shape of osteoid tissue formations, as determined histopathologically. METHODS: 20 cases of fibro-osseous lesions were investigated, comprising 5 radiographic patterns: focal (n = 3), target (n = 6), lucent (n = 4), calcification (n = 3) and multiconfluent (n = 4). Histopathological images in the central area of a full-section specimen were transformed into binary images and then into 8-bit scale images. Bone complexity and density of bone distribution were calculated and compared between patterns. RESULTS: Bone complexity score was 7384.64 for lucent, 2029.85 for focal, 2713.40 for multiconfluent, 8388.63 for calcification and 1364.27 for target pattern. The results could be broadly separated into two types: small (target, focal and multiconfluent patterns), and large (lucent and calcification patterns). Density of bone distribution was relatively low in all areas for lucent and calcification patterns, and high for focal, multiconfluent and target patterns. No significant differences in bone complexity or density of bone distribution were seen between individual patterns. CONCLUSIONS: Correlations appear to exist between image patterns from radiography and the shape of osteoid tissue on histopathology, but reorganization of the five patterns may be warranted.

Characterization of human dental pulp-derived cell lines.

AIM: To establish and characterize different types of fibroblastic cell lines derived from dental pulp tissue. METHODOLOGY: Human dental pulp tissue-derived cells were transfected with SV40 large T antigen by Lipofectamine transfection method. Geneticin (G418)-resistant cells were selected and different cell lines were established by a limiting dilution method. To characterize the lineages of cells, each clone was immunofluorescently stained by anti-fibroblast, anti-vimentin, anti-collagen type I and type III antibodies. Total RNA was extracted from each clone and subjected to a differential display experiment. RESULTS: By transfecting SV40 large T antigen, nine different cell clones were obtained. All these cell clones were positively stained by anti-fibroblast, anti-vimentin, anti-collagen type I and type III antibodies. With differential display experiment, eight different genes, the expression levels of these genes were varied amongst each cell clone, were detected. After sequencing and database search, one gene was revealed to be identical to T-cell marker, Thy-1. Thy-1 expression in dental pulp tissue was confirmed by immunohistochemical staining. CONCLUSION: Fibroblastic cell lines derived from human dental pulp tissue possessed different gene expression profiles suggesting the existence of subpopulations.

Tumour necrosis factor-alpha-mediated human polymeric immunoglobulin receptor expression is regulated by both mitogen-activated protein kinase and phosphatidylinositol-3-kinase in HT-29 cell line.

Human polymeric immunoglobulin receptor (pIgR) is present on the surface of glandular epithelium, and it plays a crucial role in the mucosal immune defence. pIgR expression in HT-29 cells is up-regulated by one of the proinflammatory cytokines, tumour necrosis factor (TNF)-alpha. However, the mechanism used by the TNF-alpha-mediated signalling pathway has not been examined exclusively. To elucidate this mechanism in detail, HT-29 cells were cotreated with TNF-alpha and mitogen-activated protein kinase kinase (MAPKK, also called MEK1) inhibitor, PD98059, and the amount of free secretory component (SC) secreted into the culture medium was measured. The amount of free SC stimulated by TNF-alpha was increased by addition of PD98059. This up-regulation occurred at the transcriptional level. The amount of SC was also up-regulated by addition of TNF-alpha with U0126, an inhibitor of MEK1 and MEK2. Nuclear factor (NF)-kappaB activity and NF-kappaB binding to the kappaB2 site localized upstream of the pIgR gene did not change after coincubation of HT-29 cells with TNF-alpha and PD98059. The expression level of pIgR by TNF-alpha was decreased by LY294002, an inhibitor of phosphatidylinositol-3-kinase (PI3K), at the transcriptional level. Extracellular signal-regulated kinase (ERK)1/2 phosphorylation and NF-kappaB binding to the kappaB2 site were not affected by LY294002 treatment. These data suggest that TNF-alpha-mediated pIgR expression is negatively regulated by ERK pathway, which is independent of NF-kappaB. In addition, decrease of SC production by Ly294002 suggests that the presence of PI3K mediated regulation of SC production.

Usefulness of cone beam computed tomography for odontogenic myxoma.

A case of odontogenic myxoma is reported as showing a cyst-like pattern with a partially thick but vague and unclear radiopaque border between the left mandibular second premolar and first molar on rotational panoramic radiography. Internal structure of the lesion displayed radiolucency with a sparse and coarse trabecular pattern. No expansion of bucco-lingual cortical bone was apparent. The radiographic pattern of odontogenic myxoma did not resemble a tennis racket-like or straight, curved and coarse septal appearance on panoramic radiography, but characteristic findings were noted on cone beam CT. Cone beam CT may prove extremely useful in clarifying detailed internal structure and the state of margins.

Co-production of vascular endothelial cadherin and inducible nitric oxide synthase by endothelial cells in periapical granuloma.

AIM: To clarify the mechanisms of inflammatory cell migration in human periapical granulomas by examining vascular endothelial (VE) cadherin and inducible nitric oxide synthase (iNOS)-producing cells. METHODOLOGY: Periapical tissues were obtained from patients during endodontic surgery and were divided into two portions. After fixing the tissues with acetone or 4% paraformaldehyde in phosphate-buffered saline, 5-microm-thick paraffin or cryostat sections were prepared, respectively. The paraffin sections of the inflamed tissues were evaluated histologically with haematoxylin-eosin stains. Cryostat sections of the tissue, diagnosed as periapical granulomas, were then examined by either immunohistochemistry using anti-human VE-cadherin or iNOS antibodies (Abs) for the characterization of infiltrating cells. In addition, co-localization of VE-cadherin and iNOS production was also analysed by two-colour immunofluorescence image analysis. RESULTS: Endothelial cells were strongly stained with iNOS Abs. Macrophages, lymphocytes, polymorphonuclear leucocytes and fibroblasts also exhibited iNOS production. These iNOS-positive cells accumulated around the blood vessels. On the other hand, VE-cadherin production was exhibited in only endothelial cells. Two-colour immunofluorescence image analysis using VE-cadherin and iNOS Abs demonstrated that iNOS-producing endothelial cells also showed VE-cadherin production. CONCLUSIONS: Vascular endothelial-cadherin produced by endothelial cells could be regulated by iNOS-producing cells in periapical granulomas and might play a pivotal role in vascular permeability.

Expression of type V secretory phospholipase A in myocardial remodelling after infarction.

AIMS: Secretory phospholipase A2 is associated with ischaemic injury in the human heart, but the distribution of type V secretory phospholipase A2 (sPLA2-V) remains unknown. The significance of sPLA2-V in myocardial infarction was investigated histopathologically. METHODS: Sequential changes in the localization of sPLA2-V and its mRNA in myocardial tissues obtained from 30 autopsied hearts were examined by immunohistochemistry and in situ hybridization and compared with those of fibronectin, vascular endothelial growth factor (VEGF), interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha, and cyclooxygenase-2 (COX-2). RESULTS: No expression of sPLA2-V was observed in normal heart, but it was promptly expressed in wavy myofibres positive for fibronectin just after the onset of infarction. sPLA2-V was subsequently expressed in ischaemic cardiomyocytes around the lesion. The expression decreased at the granulation tissue and disappeared at the chronic stage with scar formation. The distribution of the signal for sPLA2-V mRNA paralleled that of the protein. Ischaemic myocytes around the lesion expressed VEGF, IL-1beta, TNF-alpha and COX-2 at all stages. CONCLUSIONS: sPLA2-V production in myocardium is limited to the acute phase of infarction. sPLA2-V may play a dual role, acting both to remove degraded cell-membrane through cooperative activity with COX-2 in necrotic areas and to attack ischaemic myocytes around the lesion via degradation of membrane phospholipids.

New multiflorane-type triterpenoid acids from Sandoricum indicum.

Three new 12beta-hydroxymultiflorane triterpenoid acids, sandorinic acids A-C (1-3), were isolated from the stem bark of Sandoricum indicum together with five known triterpenes (4-8). The structures of 1-3 were elucidated by spectral data interpretation. Compounds 1-8 were evaluated for their inhibiting activity against several tumor cell lines and for their effects on lymphocyte proliferation.

Histopathological and immunohistological analyses in IgA deficient lymphoplasia (aly/aly) mouse.

The aly/aly mouse has a severe immunodeficiency, because it lacks peripheral lymph nodes as well as IgA and IgG immunoglobulin synthesis. In the present study, we performed histopathological and immunohistological examinations to clarify histological disorders of various immune organs in these mice. Carbon CH40 injections into the apex of the tongue confirmed the absence of submandibular lymph nodes in aly/aly mice. The thymus had a poorly constructed cortex and medulla, and the number of lymphoid follicles was clearly decreased in the spleen. No IgG- or IgA- producing cells were found in any immune organs, including the mucosal immune sites, though several IgM -producing cells were identified. Other characteristic findings included perivascular lymphocytes accumulation in the salivary glands, lungs, liver and pancreas, which caused tissues damage. These results demonstrated that the various lymphoid tissues disorders and organ-specific lymphocyte infiltration cause immuno-deficiency in the aly/aly mouse.