Analysis of proteins in human gingival crevicular fluid by mass spectrometry

Kido J, Bando M, Hiroshima Y, Iwasaka H, Yamada K, Ohgami N, Nambu T, Kataoka M, Yamamoto T, Shinohara Y, Sagawa I, Nagata T. Analysis of proteins in human gingival crevicular fluid by mass spectrometry. J Periodont Res 2012; 47: 488–499. © 2012 John Wiley & Sons A/S Background and Objective: Gingival crevicular fluid is a bodily fluid transuded from periodontal tissues into the gingival crevice and periodontal pocket, and contains many species of components. Proteins in gingival crevicular fluid have been studied as markers for periodontal diseases. Mass spectrometric analysis is used for the analyses of proteins, lipids, saccharides and metals, and expected as an approach for disease diagnosis. For better analysis of the protein components in gingival crevicular fluid, we investigated proteins in gingival crevicular fluid samples from the healthy gingival crevice and periodontal pocket using mass spectrometry. Material and Methods: Gingival crevicular fluid samples were collected from subjects who gave their informed consent and were periodontally healthy or had diseased pockets. These samples were electrophoretically separated, and each fraction on the gels was analysed by nano liquid chromatography coupled with tandem mass spectrometry. Antimicrobial peptides detected in gingival crevicular fluid were confirmed by western blotting. Results: One hundred and four proteins were detected in gingival crevicular fluid samples from both healthy sites and sites of periodontitis; 64 proteins were contained only in gingival crevicular fluid from healthy sites and 63 proteins were observed only in gingival crevicular fluid from periodontitis sites. These proteins were blood‐, cytoskeleton‐, immunity‐, inflammation‐ and lipid‐related proteins and enzymes. Some proteins, including ceruloplasmin, glycogen phosphorylase, glutathione S‐transferase, phosphoglycerate mutase, psoriasin, S100A11 and resistin, were identified for the first time in gingival crevicular fluid. Antimicrobial peptides, such as lactoferrin, α1‐antitrypsin, lipocalin, S100A7, S100A8, S100A9 and cathelicidin, were observed by mass spectrometry and western blotting. Conclusion: Multiple protein components in gingival crevicular fluid were analysed at the same time using mass spectrometry, and this approach may be useful for the diagnosis of periodontal diseases.     

Clinoidal meningioma arising from falciforme process of optic canal: Case report and review

Abstract

A 56-year-old female with an initial symptom of unilateral progressive visual disturbance presented with a clinoidal meningioma arising from a falciforme process of the optic canal. Magnetic resonance images with gadolinium-diethylenetriaminepenta-acetic acid demonstrated marked enhancement of a small mass, approximately 1 cm in diameter, roofing the right optic nerve at the intracranial opening of the optic canal. The clinoidal meningioma is rarely reported and the precise originating location of the neoplasm has not been clarified. This is the first case of clinoidal meningioma, which is confirmed by the operative observation as the origin of the neoplasm is a falciforme process of the optic canal. Magnetic resonance images with enhancement are useful for the diagnosis of such a small neoplastic lesion adjacent to bone structure and total removal without a deterioration of visual function is achieved by careful microsurgical procedure. [Neurol Res 1996; 18: 237–240]     

Effect of VA-045, a novel apovincaminic acid derivative,on closed head injury-induced neurological dysfunction in aged rats

Abstract

Objective: The strength–duration time constant (SDTC) is a measure of axonal excitability and it can provide information about Na⁺ channel function. In this study, we sought to examine the changes in the SDTCs of motor and sensory fibers of the median nerve in patients taking colchicine, which affects axoplasmic flow and may result in axonal neuropathy.

Methods and results: The SDTCs of motor and sensory fibers of 29 patients who had been taking colchicine were measured following stimulation of the right median nerve at the wrist. The results were compared with ten healthy age-matched subjects. No significant differences were found between the groups.

Conclusions: The lack of any effect on the SDTC by colchicine might have been due to the fact that axonal degeneration caused by colchicine affects the Na⁺–K⁺ ATP pump or that it affects internodal channels other than nodal channels.     

Negative pressure wound therapy for a large skin ulcer following repair of huge myeloschisis with kyphosis in a newborn

Background

In surgery of repair for spina bifida, various skin plastic techniques are sometimes necessary due to large skin defect or subsequent ulcers in cases when approximation on the midline is difficult.

Case report

A baby was born with a large skin defect due to huge lumbar myeloschisis and kyphosis, which was repaired 2 days after birth using Limberg’s skin flap at the peak of kyphosis. Skin ischemia around the tip of the flap gradually enlarged and resulted in a large skin ulcer. We performed negative pressure wound therapy (NPWT) using a Vacuum Assisted Closure (V.A.C.®) therapy system for 4 weeks which shrank the ulcer remarkably. Subsequently, a pedicle skin flap without graft was performed to cover the rest of the ulcer, which adapted well without CSF leakage postoperatively.

Conclusion

A combination treatment of NPWT and skin plastic surgery was successfully performed for a very young infant with spina bifida. NPWT could be another useful option for the treatment of ulcer following spina bifida repair surgery, though surgeons should carefully confirm that there is no CSF leakage before and during the procedure.     

ITIH4 and Gpx3 are potential biomarkers for amyotrophic lateral sclerosis

The diagnosis of amyotrophic lateral sclerosis (ALS) is difficult due to lack of definitive biomarkers. Our aim was to identify characteristic serum protein patterns that could provide candidate biomarkers for ALS. We divided mutant superoxide dismutase-1 (SOD1)^H46R rats into three groups based on disease progression: pre-symptom (90 days), onset, and end-stage. After separation of serum proteins using two-dimensional electrophoresis, we selected clear protein spots and identified two candidate proteins—inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4) and glutathione peroxidase 3 (Gpx3). The 120 kDa ITIH4 increased at the onset of the disease and the 85 kDa ITIH4, a cleaved form, at the end-stage in the sera of the SOD1^H46R rats. Expression of the 85 kDa ITIH4 was substantial in ALS compared with controls or patients with muscular dystrophy, Alzheimer diseases, or Parkinson diseases. The Gpx3 protein levels in the sera of SOD1^H46R rats were upregulated pre-symptom and gradually decreased as the disease progressed. The Gpx3 protein levels were lower in the sera of the patients with ALS than in other diseases. These results indicate that ITIH4 and Gpx3 are potential biomarkers for ALS.