Potential biomarker proteins for aspiration pneumonia detected by shotgun proteomics using buccal mucosa samples: a cross-sectional case-control study.

BACKGROUND: Aspiration pneumonia (AP), which is a major cause of death in the elderly, does present with typical symptoms in the early stages of onset, thus it is difficult to detect and treat at an early stage. In this study, we identified biomarkers that are useful for the detection of AP and focused on salivary proteins, which may be collected non-invasively. Because expectorating saliva is often difficult for elderly people, we collected salivary proteins from the buccal mucosa. METHODS: We collected samples from the buccal mucosa of six patients with AP and six control patients (no AP) in an acute-care hospital. Following protein precipitation using trichloroacetic acid and washing with acetone, the samples were analyzed by liquid chromatography and tandem mass spectrometry (LC-MS/MS). We also determined the levels of cytokines and chemokines in non-precipitated samples from buccal mucosa. RESULTS: Comparative quantitative analysis of LC-MS/MS spectra revealed 55 highly (P values < 0.10) abundant proteins with high FDR confidence (q values < 0.01) and high coverage (> 50%) in the AP group compared with the control group. Among the 55 proteins, the protein abundances of four proteins (protein S100-A7A, eukaryotic translation initiation factor 1, Serpin B4, and peptidoglycan recognition protein 1) in the AP group showed a negative correlation with the time post-onset; these proteins are promising AP biomarker candidates. In addition, the abundance of C-reactive protein (CRP) in oral samples was highly correlated with serum CRP levels, suggesting that oral CRP levels may be used as a surrogate to predict serum CRP in AP patients. A multiplex cytokine/chemokine assay revealed that MCP-1 tended to be low, indicating unresponsiveness of MCP-1 and its downstream immune pathways in AP. CONCLUSION: Our findings suggest that oral salivary proteins, which are obtained non-invasively, can be utilized for the detection of AP.

Japanese clinical practice guidelines for aspiration and pharyngeal residual assessment during eating and swallowing for nursing care.

AIM: This clinical practice guideline aims to provide and recommend methods of assessing aspiration and pharyngeal residue during eating and swallowing and methods of selecting and implementing nursing care for adults to prevent the development of aspiration pneumonia through early and appropriate management of oropharyngeal dysphagia. METHODS: In April 2018, the Japan Academy of Nursing Science established the Supervisory Committee in Nursing Care Development/Standardization Committee to develop clinical practice guidelines for aspiration and pharyngeal residual assessment during eating and swallowing for nursing care. This clinical practice guideline was developed according to the Minds Manual for Guideline Development 2017, with the aim of providing a specific pathway for nurses to determine the policy for selecting management for oropharyngeal dysphagia based on research evidence and multifaceted factors including the balance of benefits and harms and patients' values. RESULTS: Based on the 10 clinical questions related to assessment by physical assessment, the Repetitive Saliva Swallowing Test, Modified Water Swallowing Test, Food Test, cervical auscultation, observation using an ultrasound diagnostic device, and an endoscope, 10 recommendations have been developed. Eight recommendations have been evaluated as the Grading of Recommendations Assessment, Development and Evaluation (GRADE) 2C, and the other two have been evaluated as no GRADE. CONCLUSION: The first reliable clinical practice guideline has been produced from an academic nursing organization that focuses on assessment for nursing care and incorporates the latest findings.

Food bolus-forming ability predicts incidence of aspiration pneumonia in nursing home older adults: A prospective observational study.

BACKGROUND: Food boluses in the pharynx without enough bolus formation sometimes cause aspiration among older adults; however, the relationship between food bolus-forming ability and incidence of aspiration pneumonia is unclear. OBJECTIVE: To investigate the relationship between food bolus-forming ability and incidence of aspiration pneumonia by evaluating the condition of chew-swallow managing food transported into the pharynx. METHODS: A prospective observational study conducted in a nursing home for the elderly between April 2016 and February 2018. Seventy-three residents who swallowed thickened liquids and consistent boluses without aspiration were included. Food boluses were graded into three categories in the pharynx using videoendoscopic evaluation. Boluses that retained their original shape were defined as Grade 1. A mixture of large and small boluses was defined as Grade 2. Boluses that had completely transformed into a paste were defined as Grade 3. The relationship between the bolus formation grade and incidence of aspiration pneumonia over 6-month follow-up was investigated. RESULTS: Seventeen residents developed aspiration pneumonia. The incidence rate of aspiration pneumonia was highest among residents with Grade 1 boluses, at an incidence rate of .187 cases per person-month (95% confidence interval (CI): 0.097-0.359). Cox regression showed residents with Grade 1 boluses had a hazard ratio of 4.548 (1.393-14.85) for incidence of aspiration pneumonia compared with residents who had Grade 2 or 3 boluses. CONCLUSION: Insufficient food bolus-forming ability predicted high incidence of aspiration pneumonia. Healthcare professionals should evaluate the food bolus-forming ability, as well as swallowing function, to prevent aspiration pneumonia.

Towards a beyond 1 GHz solid-state nuclear magnetic resonance: external lock operation in an external current mode for a 500 MHz nuclear magnetic resonance.

Achieving a higher magnetic field is important for solid-state nuclear magnetic resonance (NMR). But a conventional low temperature superconducting (LTS) magnet cannot exceed 1 GHz (23.5 T) due to the critical magnetic field. Thus, we started a project to replace the Nb(3)Sn innermost coil of an existing 920 MHz NMR (21.6 T) with a Bi-2223 high temperature superconducting (HTS) innermost coil. Unfortunately, the HTS magnet cannot be operated in persistent current mode; an external dc power supply is required to operate the NMR magnet, causing magnetic field fluctuations. These fluctuations can be stabilized by a field-frequency lock system based on an external NMR detection coil. We demonstrate here such a field-frequency lock system in a 500 MHz LTS NMR magnet operated in an external current mode. The system uses a (7)Li sample in a microcoil as external NMR detection system. The required field compensation is calculated from the frequency of the FID as measured with a frequency counter. The system detects the FID signal, determining the FID frequency, and calculates the required compensation coil current to stabilize the sample magnetic field. The magnetic field was stabilized at 0.05 ppm∕3 h for magnetic field fluctuations of around 10 ppm. This method is especially effective for a magnet with large magnetic field fluctuations. The magnetic field of the compensation coil is relatively inhomogeneous in these cases and the inhomogeneity of the compensation coil can be taken into account.

Significance of stromal desmoplasia and myofibroblast appearance at the invasive front in squamous cell carcinoma of the oral cavity.

BACKGROUND: Tumor invasion involves complex interactions between tumor and stromal cells. We examined the extent of connective tissue in the tumor stroma and whether myofibroblasts play a role in assisting cancer invasion and metastasis. METHODS: Biopsy materials from 84 patients with oral squamous cell carcinoma (SCC) were used. We compared data from intrastromal collagen fibers using Azan staining, immunohistochemical identification of myofibroblasts by cytoskeletal markers, alpha-smooth muscle actin, vimentin, desmin, and clinicopathological parameters. Clinical outcome was compared by 5-year survival rate. RESULTS: There were high levels of stromal collagen fibers in invasive tumors. Myofibroblast appearance increased with increasing tumor invasiveness. Lymph node metastasis occurred more frequently in the myofibroblast-positive group, and the survival rate was significantly poorer in this group. CONCLUSIONS: Fibrous stroma in SCC appeared to have a desmoplastic response. However, an independent invasive mechanism may regulate the stroma, with tumor desmoplasia occurring in highly developed, invasive tumors.

Integrin expression levels correlate with invasion, metastasis and prognosis of oral squamous cell carcinoma.

The present study evaluated the relationship between alpha 3, alpha 6A, and beta 1 integrin expression in cancer cells at the invasive front of oral squamous cell carcinoma (OSCC) and survival rates, as well as the clinical and pathological characteristics. Sections of 100 specimens of primary OSCC were immunostained to assess alpha 3, alpha 6A, and beta 1 integrin expression in cancer cells at the invasive front. OSCC patients with higher expression levels of alpha 3, alpha 6A, and beta 1 integrin had significantly better prognosis than those with lower expression levels (median survival at low vs. high expression levels: alpha 3, 37.1 months vs. 55.7 months; alpha 6A , 38.3 months vs. 47.9 months; and beta 1, 26.1 months vs. 46.1 months) (P < 0.05). In addition, beta 1 integrin expression showed the highest correlation with clinical and pathological characteristics. This study concludes that alpha 3, alpha 6A, and beta 1 integrin expression in cancer cells at the invasive front are related to the mode of invasion and prognosis in OSCC.

Predictive value of measuring p53 labeling index at the invasive front of oral squamous cell carcinomas.

Many studies have revealed the frequency of p53 abnormalities in oral cancer. However, it reports only on the relation between clinicopathological findings and p53 expression, and there is no study to examine the relation to the p53 labeling index (p53-LI). The purposes of this study were to examine the correlation between p53 labeling index (p53-LI) at the invasive front of oral squamous cell carcinomas (OSCC) and clinicopathological findings by immunohistochemical staining, and to evaluate clinical significance of measuring p53-LI at the invasive front of OSCC. Sixty-six biopsy specimens of OSCC were randomly selected. Patient age, gender, primary sites, T category, N category, degree of differentiation and mode of cancer invasion were analyzed. p53 expression did not correlate significantly with the clinical findings. However, significant differences were found between p53-LI and the degree of cell differentiation (p < 0.05). The p53-LI of high-grade invasive tumors was significantly larger than that of low-grade invasive tumors (p < 0.05). The overall survival rate (OS) among low-scoring p53-LI cases was 75.5% whereas that for high-scoring p53-LI cases was 40.6%. The disease-free survival rate (DFS) among low-scoring p53-LI cases was 39.5% whereas that for high-scoring p53-LI cases was 76.1%. Patients with low-scoring p53-LI had a significantly worse prognosis than those with among high-scoring p53-LI (p < 0.05). Consequently, the measurement of p53-LI at the invasive front of OSCC is significant as one of the indicators of prognosis.

Expression of bone morphogenetic protein 2 and fibroblast growth factor 2 during bone regeneration using different implant materials as an onlay bone graft in rabbit mandibles.

OBJECTIVES: The purpose of this study was to histologically and immunohistochemically evaluate bone regeneration using 3 different implant materials in rabbit mandibles and to compare the bone regenerative capability of these materials in an animal model. STUDY DESIGN: Adult male Japanese white rabbits (n = 48; 12-16 wks old; 2.5-3.0 kg) were divided into 4 groups, consisting of 12 animals each. The implant materials were beta-tricalcium phosphate (beta-TCP), autologous bone derived from the radius, and recombinant human bone morphogenetic protein 2 (rhBMP-2) with polylactic acid/polyglycolic acid copolymer and gelatin sponge (PGS) complex. After incising along the inferior border of the mandible, the materials were implanted as only grafts and covered by titanium mesh with screws. No material was implanted into the control group. The rabbits were killed at 2, 4, 8, 12, and 24 wks postoperatively, and formalin-fixed specimens containing titanium mesh were embedded in acrylic resin. The specimens were stained with hematoxylin and eosin. For immunohistochemical analysis, the specimens were treated with BMP-2 and fibroblast growth factor 2 (FGF-2) antibodies. Finally, they were examined microscopically. RESULTS: The autologous bone induced substantially more new bone formation compared with beta-TCP at 4 wks postoperatively. However, rhBMP-2/PGS induced new bone formation at 8 wks postoperatively. No growth of bony tissue was observed in the control group at any period. In the autologous bone and rhBMP-2/PGS groups, both BMP-2 and FGF-2 were observed later in the beta-TCP group than in other groups. CONCLUSION: This study suggests that autologous bone as well as rhBMP-2/PGS implants induce expression of both BMP-2 and FGF-2 specifically at the operated sites, even at early stages.

Correlation of basic fibroblast growth factor expression with the invasion and the prognosis of oral squamous cell carcinoma.

BACKGROUND: The aim of this study was to evaluate the relationship between the expression of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor-1 (FGFR-1) in cancer cells and fibroblasts at the invasive front of oral squamous cell carcinoma (OSCC), and the pathologic and clinical characteristics. METHODS: Sections of 61 biopsy specimens of primary OSCC were immunostained to assess the expression of bFGF and FGFR-1 in cancer cells and fibroblasts at the invasive front. RESULTS: The bFGF and FGFR-1 expressions in the cancer cells were evident in all specimens, whilst, in fibroblasts, they were detected in 41 (67%) of 61 specimens. These expressions in the fibroblasts occurred notably more often in high-invasive OSCC specimens than low-invasive OSCC specimens. The prevalence of bFGF and FGFR-1 expressions in cases with lymph node metastasis was significantly higher (P < 0.05) than in cases without metastasis. Moreover, these expressions were well correlated with patient prognosis. CONCLUSION: This study concludes that bFGF and FGFR-1 expressions in fibroblasts at the invasive front are linked to the mode of invasion and the prognosis in OSCC.

Influences of angiogenesis and lymphangiogenesis on cancerous invasion in experimentally induced tongue carcinoma.

BACKGROUND: Although it is clear that dissemination via the blood system involves angiogenesis, it is uncertain whether tumors also induce lymphangiogenesis or simply invade existing peritumoral vessels. The purpose of this study was to elucidate changes in tumor blood and lymph vessels in cases involving the invasion of squamous cell carcinoma in the oral cavity, and its significance. Blood and lymph vessels densities in tongue carcinomas induced in hamsters were investigated. METHODS: Tongue cancer was induced by abrading the right margin of the tongue of each hamster with an endodontic barbed broach and subsequently applying 1.0% 9,10-dimenthl-1,2-benzanthracene (DMBA) dissolved in acetone, three times a week, at the same site. Fresh frozen sections were prepared and blood vessels stained blue by perfusion with Coomassie Brilliant Blue and lymph vessels stained brown for 5'-nucleotidase. The effects on the blood vessels and lymph vessels were observed. RESULTS: The results showed that blood and lymph vessel densities were greater in the advanced carcinoma tissues than in normal tissue. These were compared in terms of the mode of cancer invasion. As tumor invasion progressed, the blood vessel density decreased but lymph vessel density tended to be higher in high-degree tumor invasion than in low-degree tumor invasion. The expression of vascular endothelial growth factor-C was seen more frequently as tumor invasion progressed. CONCLUSIONS: The present findings indicated that angiogenesis and lymphangiogenesis are affected by cancerous invasion.