Motowo Nabeta

Identification of anti-α-enolase autoantibody as a novel serum marker for endometriosis

Diagnosis of endometriosis needs invasive maneuvers. New serum marker that possesses both high sensitivity and high specificity has long been desired. To establish novel serum marker for endometriosis, serum autoantibodies (autoAbs) were investigated using proteomic approach. AutoAbs in sera of endometriotic patients and healthy controls were analyzed using a mesothelial cell line, 2‐DE and Western blotting. Proteins in reacted spots were identified using MALDI TOF‐MS with MASCOT analysis. ELISAs were established using recombinant proteins and autoAb‐titers were estimated in sera of endometriotic patients, disease and healthy controls. Several autoAbs were identified. Anti‐α‐enolase (Eno1)‐autoAb levels in endometriotic patients were significantly elevated compared with both healthy and disease controls. Sensitivity and specificity of serum anti‐Eno1‐autoAb was nearly comparable to serum CA125. When anti‐Eno1‐autoAb and CA125 assays were combined, diagnostic sensitivity and accuracy improved. Serum anti‐Eno1‐autoAb can be a new serum endometriotic marker and it is useful as a supplement assay for CA125. This study validates further clinical evaluation of this novel marker.

Serum autoantibody to sideroflexin 3 as a novel tumor marker for oral squamous cell carcinoma

The purpose of this study is to establish a tumor marker that can be applied for the early detection and follow‐up of oral cancer patients. Employing the proteomic approach using MALDI TOF‐MS, 2‐DE, patients sera and culturing cell lines, the serum autoantibodies (autoAbs) were screened and the serum levels were estimated by ELISA. Targeting the tumor cell invasion into the surrounding stromal tissues, MRC‐5 human fibroblasts were employed as the target cells and a mitochondrial membrane protein, sideroflexin 3 (SFXN3), was identified. The serum anti‐SFXN3‐autoAb levels elevated in patients with the oral squamous cell carcinoma significantly: with 77% sensitivity and 89% specificity against control samples. The serum anti‐SFXN3‐autoAb levels were mildly correlated with the primary tumor sizes, however, the levels were slightly highly elevated in T1 early cancer. An immunohistochemical analysis revealed that the SFXN3 protein is expressed in the stromal fibroblasts between the caner nests and also in the basal layer of the squamous epithelium. Changes in the serum anti‐SFXN3‐autoAb levels after therapy correlated with the clinical tumor burden. These findings demonstrated that the serum anti‐SFXN3‐autoAb is worthy of clinical evaluation as a potentially of the novel tumor maker for the early detection of oral squamous cell carcinoma.

Identification of anti-syntaxin 5 autoantibody as a novel serum marker of endometriosis

The sensitivity and specificity of CA125, as a sole serum marker of endometriosis, are not high enough for routine clinical assessment. To explore new markers for the diagnosis of endometriosis, serum autoantibodies in endometriotic patients were investigated employing a fibroblast cell line, two-dimensional (2D) gel electrophoresis and Western blotting. Proteins reacting with serum autoantibodies by Western blotting were identified using MASCOT analysis. ELISAs were then prepared using recombinant proteins and titers of serum autoantibodies were determined in the endometriotic patients, disease controls, and healthy subjects. Among the autoantibodies identified, anti-syntaxin 5 (STX5) autoantibody levels were significantly elevated in endometriotic patients. Sensitivity (53.6%) and accuracy (72.2%) of the serum anti-STX5 autoantibody assay were better than those of serum CA125 levels (36.2% and 62.9%, respectively) for diagnosis. The sensitivity of anti-STX5 autoantibody was remarkably high in Stage II (80.0%) compared with that of CA125 (40.0%). A combination assay of anti-STX5 autoantibody with CA125 improved the overall sensitivity to 69.6%. We conclude that serum anti-STX5 autoantibody, which was discovered by a proteomic approach, is a potential new serum marker for the diagnosis of endometriosis. This initial study now requires validation by further clinical evaluation.

Serum anti-PDIK1L autoantibody as a novel marker for endometriosis

OBJECTIVE To establish a novel serum marker for endometriosis, serum autoantibodies (autoAbs) were investigated using a proteomic approach. DESIGN Retrospective study. SETTING Departments of Molecular Pathology and Obstetrics and Gynecology in Ehime University and University Hospital. PATIENT(S) Sixty-nine patients with endometriosis, 38 disease control patients without endometriosis, and 44 healthy volunteers. INTERVENTION(S) Autoantibodies in sera of endometriotic patients and healthy controls were investigated using a human fibroblast cell line, two-dimensional gel electrophoresis, and Western blotting. Proteins in reacted spots were identified using MALDI time of flight mass spectrometry with MASCOT analysis. ELISAs were established using recombinant proteins, and autoAb-titers were estimated in sera of endometriotic patients and controls. MAIN OUTCOME MEASURE(S) Identification of serum autoAb useful for diagnosis of endometriosis. RESULT(S) Several autoAbs were identified. ELISAs were established and serum autoAb titers were estimated. Among those identified, anti-PDIK1L-autoAb levels were significantly elevated in endometriotic patients. Sensitivity (59.4%) and accuracy (72.8%) of serum anti-PDIK1L-autoAb assay were better than those of serum CA125 levels (36.2% and 62.9%, respectively) in diagnosis of endometriosis. Additionally, anti-PDIK1L-autoAb could detect endometriotic patients in early stages. CONCLUSION(S) Serum anti-PDIK1L-autoAb can be a new serum marker for the diagnosis of endometriosis. This study validates further clinical evaluation of this novel marker.

Sugar expression in the mucosae of the canine uterus and vagina during the oestrous cycle and with pyometra.

The pathogenesis of canine pyometra is still unclear, but bacterial infection of the endometrium, mediated by bacterial lectins, is suspected to induce pyometra. The aim of this study was to investigate sugar expression in the mucosae of the uterus and vagina of healthy dogs with normal oestrous cycles and in dogs with pyometra, using a panel of lectins to investigate the pathogenesis of pyometra. In dogs with pyometra, the uterine and vaginal mucosae were positive for lectins that selectively bind to glucose or mannose, especially during days 7-10 and 30-40 of dioestrus. These results suggest that temporal changes in sugar expression in the uterus and vagina present an opportunity for pathogens to infect the endometrium, causing pyometra.