Error-suppression mechanism of PCR by blocker strands.

The polymerase chain reaction (PCR) is a central technique in biotechnology. Its ability to amplify a specific target region of a DNA sequence has led to prominent applications, including virus tests, DNA sequencing, genotyping, and genome cloning. These applications rely on the specificity of the primer hybridization and therefore require effective suppression of hybridization errors. A simple and effective method to achieve that is to add blocker strands, also called clamps, to the PCR mixture. These strands bind to the unwanted target sequence, thereby blocking the primer mishybridization. Because of its simplicity, this method is applicable to a broad nucleic-acid-based biotechnology. However, the precise mechanism by which blocker strands suppress PCR errors remains to be understood, limiting the applicability of this technique. Here, we combine experiments and theoretical modeling to reveal this mechanism. We find that the blocker strands both energetically destabilize the mishybridized complex and sculpt a kinetic barrier to suppress mishybridization. This combination of energetic and kinetic biasing extends the viable range of annealing temperatures, which reduces design constraint of the primer sequence and extends the applicability of PCR.

Influence of Liquid on the Tooth Surface on the Accuracy of Intraoral Scanners: An In Vitro Study.

PURPOSE: To assess the influence of liquid attached on the tooth surfaces on the accuracy (trueness and precision) of intraoral scanners and the effectiveness of the drying method (using compression air) to exclude the influence of liquid on the scanning results. MATERIALS AND METHODS: A mandibular jaw model was scanned using an industrial computed tomography scanner to obtain a reference model. A scanning platform was designed to simulate three specific tooth surface states (dry, wet, blow-dry). Two kinds of liquids (ultra-pure water and artificial saliva) were used for the test. Two intraoral scanners (Trios 3 and Primescan) were used to scan the mandibular jaw model 10 times under each condition. All scanning data were processed and analyzed using dedicated software (Geomagic Control 2015). Trueness and precision comparison were conducted within the 12 groups of 3D models divided based on different intraoral scanners and liquids used under each condition. The root mean square (RMS) value was used to indicate the difference between the aligned virtual models. The color maps were used to evaluate and observe the deviation distribution patterns. The 3-way ANOVA (condition, intraoral scanner, liquid) followed by the Tukey test were used to assess precision and trueness. The level of significance was set at 0.05. RESULTS: The mean RMS values obtained from wet condition were significantly higher than those of the dry and blow-dry condition (p < 0.001, F = 64.033 for trueness and F = 54.866 for precision), which indicates less accurate trueness and precision for wet condition. For two different types of liquids, the mean RMS value was not significantly different on trueness and precision. The deviations caused by liquid were positive and mainly distributed in the pits and fissures of the occlusal surface of posterior teeth, the interproximal area of the teeth, and the margin of the abutments. CONCLUSIONS: Liquid on the tooth surface could affect intraoral scanning accuracy. Blow-drying with a three-way syringe can reduce scanning errors.

Genetic and epigenetic analyses of panic disorder in the post-GWAS era.

Panic disorder (PD) is a common and debilitating neuropsychiatric disorder characterized by panic attacks coupled with excessive anxiety. Both genetic factors and environmental factors play an important role in PD pathogenesis and response to treatment. However, PD is clinically heterogeneous and genetically complex, and the exact genetic or environmental causes of this disorder remain unclear. Various approaches for detecting disease-causing genes have recently been made available. In particular, genome-wide association studies (GWAS) have attracted attention for the identification of disease-associated loci of multifactorial disorders. This review introduces GWAS of PD, followed by a discussion about the limitations of GWAS and the major challenges facing geneticists in the post-GWAS era. Alternative strategies to address these challenges are then proposed, such as epigenome-wide association studies (EWAS) and rare variant association studies (RVAS) using next-generation sequencing. To date, however, few reports have described these analyses, and the evidence remains insufficient to confidently identify or exclude rare variants or epigenetic changes in PD. Further analyses are therefore required, using sample sizes in the tens of thousands, extensive functional annotations, and highly targeted hypothesis testing.

Genetic and environmental factors of schizophrenia and autism spectrum disorder: insights from twin studies.

Twin studies of psychiatric disorders such as schizophrenia and autism spectrum disorder have employed epidemiological approaches that determine heritability by comparing the concordance rate between monozygotic twins (MZs) and dizygotic twins. The basis for these studies is that MZs share 100% of their genetic information. Recently, biological studies based on molecular methods are now being increasingly applied to examine the differences between MZs discordance for psychiatric disorders to unravel their possible causes. Although recent advances in next-generation sequencing have increased the accuracy of this line of research, there has been greater emphasis placed on epigenetic changes versus DNA sequence changes as the probable cause of discordant psychiatric disorders in MZs. Since the epigenetic status differs in each tissue type, in addition to the DNA from the peripheral blood, studies using DNA from nerve cells induced from postmortem brains or induced pluripotent stem cells are being carried out. Although it was originally thought that epigenetic changes occurred as a result of environmental factors, and thus were not transmittable, it is now known that such changes might possibly be transmitted between generations. Therefore, the potential possible effects of intestinal flora inside the body are currently being investigated as a cause of discordance in MZs. As a result, twin studies of psychiatric disorders are greatly contributing to the elucidation of genetic and environmental factors in the etiology of psychiatric conditions.

New Class of Amphiphiles Designed for Use in Water-in-Supercritical CO2 Microemulsions.

Water-in-supercritical CO2 microemulsions formed using the hybrid F-H surfactant sodium 1-oxo-1-[4-(perfluorohexyl)phenyl]hexane-2-sulfonate, FC6-HC4, have recently been shown to have the highest water-solubilizing power ever reported. FC6-HC4 demonstrated the ability to outperform not only other surfactants but also other FCm-HCn analogues containing different fluorocarbon and hydrocarbon chain lengths (Sagisaka, M. et al. Langmuir 2015, 31, 7479-7487). With the aim of clarifying the key structural features of this surfactant, this study examined the phase behavior and water/supercritical CO2 aggregate formation of 1-oxo-1-[4-(perfluorohexyl)phenyl]hexane (Nohead FC6-HC4), which is an FC6-HC4 analogue but now, interestingly, without the sulfonate headgroup. Surprisingly, Nohead FC6-HC4, which would not normally be identified as a classic surfactant, yielded transparent single-phase W/CO2 microemulsions with polar cores able to solubilize a water-soluble dye, even at pressures and temperatures so low as to approach the critical point of CO2 (e.g., ∼100 bar at 35 °C). High-pressure small-angle scattering (SANS) measurements revealed the transparent phases to consist of ellipsoidal nanodroplets of water. The morphology of these droplets was shown to be dependent on the pressure, Nohead FC6-HC4 concentration, and water-to-surfactant molar ratio. Despite having almost the same structure as Nohead FC6-HC4, analogues containing both shorter and longer hydrocarbons were unable to form W/CO2 microemulsion droplets. This shows the importance of the role of the hydrocarbon chain in the stabilization of W/CO2 microemulsions. A detailed examination of the mechanism of Nohead FC6-HC4 adsorption onto the water surface suggests that the hexanoyl group protrudes into the aqueous core, allowing for association between the carbonyl group and water.

Vasodilatory mechanisms of unoprostone isopropyl in isolated porcine retinal arterioles.

PURPOSE: To clarify the vasodilatory mechanism of unoprostone isopropyl (UI), we examined its effects on the retinal microvascular diameter to determine the dependence on the endothelium and/or smooth muscle to reveal the signaling mechanisms involved in this vasomotor activity. METHODS: Porcine retinal arterioles were isolated, cannulated, and pressurized without flow in vitro. Video microscopic techniques recorded the diametric responses to UI. RESULTS: The retinal arterioles dilated in response to UI in a dose-dependent (100 pM-10 µM) manner. The nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) inhibited UI-induced vasodilation. The large-conductance Ca2+-activated K channel (BKCa channel) blocker iberiotoxin also inhibited UI-induced vasodilation. The residual vasodilation after L-NAME was eliminated with co-administration of iberiotoxin. CONCLUSIONS: UI elicits dilation of the retinal arterioles mediated by NO release and BKCa channel activation.

Effect of Fluorocarbon and Hydrocarbon Chain Lengths in Hybrid Surfactants for Supercritical CO2.

Hybrid surfactants containing both fluorocarbon (FC) and hydrocarbon (HC) chains have recently been shown to solubilize water and form elongated reversed micelles in supercritical CO2. To clarify the most effective FC and HC chain lengths, the aggregation behavior and interfacial properties of hybrid surfactants FCm-HCn (FC length m/HC length n = 4/2, 4/4, 6/2, 6/4, 6/5, 6/6, and 6/8) were examined in W/CO2 mixtures as functions of pressure, temperature, and water-to-surfactant molar ratio (W0). The solubilizing power of hybrid surfactants for W/CO2 microemulsions was strongly affected by not only the FC length but also by that of the HC. Although the surfactants having short FC and/or HC tails (namely, m/n = 4/2, 4/4, and 6/2) did not dissolve in supercritical CO2 (even at ∼17 mM, ≤400 bar, temperature ≤ 75 °C, and W0 = 0-40), the other hybrid surfactants were able to yield transparent single-phase W/CO2 mixtures identified as microemulsions. The solubilizing power of FC6-HCm surfactants reached a maximum (W0 ∼ 80 at 45 °C and 350 bar) with a hydrocarbon length, m, of 4. The W0 value of 80 is the highest for a HC-FC hybrid surfactant, matching the highest value reported for a FC surfactant which contained more FC groups. High-pressure small-angle neutron scattering measurements from FCm-HCn/D2O/CO2 microemulsions were consistent with growth of the microemulsion droplets with increasing W0. In addition, not only spherical reversed micelles but also nonspherical assemblies (rodlike or ellipsoidal) were found for the systems with FC6-HCn (n = 4-6). At fixed surfactant concentration and W0 (17 mM and W0 = 20), the longest reversed micelles were obtained for FC6-HC6 where a mean aspect ratio of 6.3 was calculated for the aqueous cores.

Genomic structural variation in affective, anxiety, and stress-related disorders.

Anxiety and stress-related disorders are common and debilitating mental diseases. Genetic factors as well as environmental factors and life events contribute to their pathogenesis and partly mediate treatment response. However, these disorders are clinically heterogeneous, genetically complex, and the exact genetic causes are still unclear. Recently, some evidence has emerged for structural variation including copy number variants and small deletions/duplications to be associated with mental disorders. Here, the current state of knowledge on the role of genomic structural variation in affective, anxiety and stress-related disorders is reviewed, followed by a critical discussion of present methods to detect structural changes, future directions, and clinical implications including a potential role in personalized medicine.

Role of Ca2+ -dependent and Ca2+ -sensitive mechanisms in sphingosine 1-phosphate-induced constriction of isolated porcine retinal arterioles in vitro.

Although sphingosine 1-phosphate (S1P), a bioactive lipid derived from activated platelets, has a variety of physiologic effects on vessels, no reports have described the effect of S1P on the retinal circulation. We examined the effect and underlying mechanism of the vasomotor action of S1P on porcine retinal arterioles. The porcine retinal arterioles were isolated, cannulated, and pressurized without flow for in vitro study. S1P-induced diameter changes were recorded using videomicroscopic techniques. S1P elicited concentration-dependent (1 nM-10 µM) vasoconstriction of the retinal arterioles that was abolished by the S1P receptor 2 (S1PR2) antagonist JTE-013. S1P-induced vasoconstriction was abolished by the Rho kinase (ROCK) inhibitor H-1152 and was inhibited partly by the protein kinase C (PKC) inhibitor Gö-6983. The inhibition of phospholipase C by U73122 and L-type voltage-operated calcium channels (L-VOCCs) by nifedipine inhibited S1P-induced vasoconstriction; a combination of both inhibitors abolished S1P-induced vasoconstriction. Furthermore, inhibition of myosin light chain kinase (MLCK) by ML-9 significantly blocked S1P-induced vasoconstriction; further coadministration of ML-9 with H-1152 or Gö-6983 abolished S1P-induced vasoconstriction. The current data suggest that S1P elicits vasoconstriction of the retinal arterioles via S1PR2 in vascular smooth muscle cells and this vasoconstriction may be mediated by the Ca2+ -sensitive pathway via activation of PKC leading to activation of ROCK and the Ca2+ -dependent pathway via activation of L-VOCCs resulting in activation of MLCK.

Abrogation of ER stress-induced apoptosis of alveolar epithelial cells by angiotensin 1-7.

Earlier work showed that apoptosis of alveolar epithelial cells (AECs) in response to endogenous or xenobiotic factors is regulated by autocrine generation of angiotensin (ANG) II and its counterregulatory peptide ANG1-7. Mutations in surfactant protein C (SP-C) induce endoplasmic reticulum (ER) stress and apoptosis in AECs and cause lung fibrosis. This study tested the hypothesis that ER stress-induced apoptosis of AECs might also be regulated by the autocrine ANGII/ANG1-7 system of AECs. ER stress was induced in A549 cells or primary cultures of human AECs with the proteasome inhibitor MG132 or the SP-C BRICHOS domain mutant G100S. ER stress activated the ANGII-generating enzyme cathepsin D and simultaneously decreased the ANGII-degrading enzyme ACE-2, which normally generates the antiapoptotic peptide ANG1-7. TAPI-2, an inhibitor of ADAM17/TACE, significantly reduced both the activation of cathepsin D and the loss of ACE-2. Apoptosis of AECs induced by ER stress was measured by assays of mitochondrial function, JNK activation, caspase activation, and nuclear fragmentation. Apoptosis induced by either MG132 or the SP-C BRICHOS mutant G100S was significantly inhibited by the ANG receptor blocker saralasin and was completely abrogated by ANG1-7. Inhibition by ANG1-7 was blocked by the specific mas antagonist A779. These data show that ER stress-induced apoptosis is mediated by the autocrine ANGII/ANG1-7 system in human AECs and demonstrate effective blockade of SP-C mutation-induced apoptosis by ANG1-7. They also suggest that therapeutic strategies aimed at administering ANG1-7 or stimulating ACE-2 may hold potential for the management of ER stress-induced fibrotic lung disorders.

Low shear stress up-regulation of proinflammatory gene expression in human retinal microvascular endothelial cells.

The vascular endothelium responds to shear stress generated by blood flow and changes functions to regulate blood flow and maintain tissue homeostasis. Recently, we found that arteriolar high shear stress leads to increased expression of vasodilatory and antithrombotic genes in human retinal microvascular endothelial cells (HRMECs). However, it is unknown whether low shear stress, which is induced by hypoperfusion particularly in the retinal venules where leukocyte-endothelial interactions mainly occur, affects the retinal endothelial function. We studied the effect of low shear stress on proinflammatory gene expression in HRMECs. The cells were cultured on glass plates and exposed to laminar shear stresses of 0 (static), 1.5 (relatively low flow), and 15 dyne/cm(2) (relatively high flow) for 24 h using parallel plate-type flow-loading devices. The mRNA expressions of adhesion molecules, cytokines and chemokines, and procoagulant factors were evaluated using real-time reverse-transcription polymerase chain reaction. HRMECs exposed to 1.5 dyne/cm(2) significantly up-regulated the mRNA expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin. The cells exposed to 1.5 dyne/cm(2) of stress also had increased cytokine/chemokine mRNA expression, i.e., interleukin (IL)-6, IL-8, platelet-derived growth factor-B, and monocyte chemotactic protein-1. Procoagulant factors, i.e., tissue factor and plasminogen activator inhibitor-1 mRNA, increased significantly with exposure to 1.5 dyne/cm(2) of stress. Our results showed that relatively low shear stress causes up-regulation of proinflammatory genes in HRMECs, suggesting that decreased shear stress due to vascular hypoperfusion might change the phenotypic characterization of the retinal vascular endothelium and be associated with leukocyte-endothelial interactions.

Nanostructures in water-in-CO2 microemulsions stabilized by double-chain fluorocarbon solubilizers.

High-pressure small-angle neutron scattering (HP-SANS) studies were conducted to investigate nanostructures and interfacial properties of water-in-supercritical CO2 (W/CO2) microemulsions with double-fluorocarbon-tail anionic surfactants, having different fluorocarbon chain lengths and linking groups (glutarate or succinate). At constant pressure and temperature, the microemulsion aqueous cores were found to swell with an increase in water-to-surfactant ratio, W0, until their solubilizing capacities were reached. Surfactants with fluorocarbon chain lengths of n = 4, 6, and 8 formed spherical reversed micelles in supercritical CO2 even at W0 over the solubilizing powers as determined by phase behavior studies, suggesting formation of Winsor-IV W/CO2 microemulsions and then Winsor-II W/CO2 microemulsions. On the other hand, a short C2 chain fluorocarbon surfactant analogue displayed a transition from Winsor-IV microemulsions to lamellar liquid crystals at W0 = 25. Critical packing parameters and aggregation numbers were calculated by using area per headgroup, shell thickness, the core/shell radii determined from SANS data analysis: these parameters were used to help understand differences in aggregation behavior and solubilizing power in CO2. Increasing the microemulsion water loading led the critical packing parameter to decrease to ~1.3 and the aggregation number to increase to >90. Although these parameters were comparable between glutarate and succinate surfactants with the same fluorocarbon chain, decreasing the fluorocarbon chain length n reduced the critical packing parameter. At the same time, reducing chain length to 2 reduced negative interfacial curvature, favoring planar structures, as demonstrated by generation of lamellar liquid crystal phases.

Optical impression method to measure three-dimensional position and orientation of dental implants using an optical tracker.

OBJECTIVES: The aim of this study was to devise an optical impression method that could make impressions of dental implants accurately and rapidly. MATERIALS AND METHODS: Four paper markers (4 × 3 mm, 8 × 6 mm, 16 × 12 mm, and 24 × 18 mm) and one titanium marker (8 × 6 mm) were prepared to determine the measuring accuracy of the three-dimensional optical tracker. For a proposed and conventional impression taking method, we compared the reproduction accuracies of the positions and orientations of dental implants and the times to obtain impressions. Finally, we fabricated computer-aided designing (CAD)/computer-aided manufacturing (CAM) superstructure frameworks to determine the adaptation accuracy. RESULTS: The 8 × 6-mm titanium marker was optimal among the prepared markers. Dental implants made by the proposed and conventional impression taking methods had measurement errors of 71 ± 31 µm and 32 ± 18 µm, respectively. The proposed method took a significantly shorter time to obtain an impression than did the conventional method. The connection between the CAD/CAM superstructure frameworks and four implant analogs had uplifts of 55 ± 10 µm, 94 ± 35 µm, 2 ± 1 µm, and 66 ± 3 µm. CONCLUSION: Our proposed method and fabricated titanium markers enabled us to measure the positions and orientations of dental implants both accurately and rapidly. We then used the reproducible measurement results for the positions and orientations of the dental implants to fabricate CAD/CAM superstructure frameworks within an acceptable accuracy range.

Glutathione depletion results in S-nitrosylation of protein disulfide isomerase in neuroblastoma cells.

AIMS: Protein disulfide isomerase (PDI) is an essential enzyme involved in oxidative protein folding. PDI is S-nitrosylated in the brains of Alzheimer's disease patients, and S-nitrosylated PDI is considered one of main causes of Alzheimer's disease. However, the mechanisms underlying PDI S-nitrosylation have not yet been elucidated. Because glutathione (GSH) depletion is a pathological feature of Alzheimer's disease, we investigated the effect of GSH depletion on the S-nitrosylation level of PDI. MAIN METHODS: SH-SY5Y cells, which is a human derived neuroblastoma cells, were used in this study. Glutamate and buthionine sulfoximine (BSO) were used as GSH depletors. S-nitrosylated PDI was detected by biotin-switch assay. KEY FINDINGS: GSH depletion by glutamate, a cystine/glutamate antiporter xCT inhibitor, increased S-nitrosylated PDI at C343 in SH-SY5Y cells, and induced IRE1α phosphorylation. BSO, a γ-glutamylcysteine synthetase inhibitor, also increased S-nitrosylated PDI and phosphorylated IRE1α upon GSH depletion. Because S-nitrosylated PDI at C343 is stable in neuro cells, S-nitrosylated PDI by GSH depletion progresses to neurodegeneration by the induction of endoplasmic reticulum stress via phosphorylated IRE1α signaling from the early to late stage. Furthermore, treatment with neohesperidin, but not N-acetylcysteine (NAC), improved PDI S-nitrosylation level in GSH-depleted SH-SY5Y cells because nitrosylated compound of NAC induces PDI S-nitrosylation. SIGNIFICANCE: The results of our study provide a new insight into the mechanisms of neurodegeneration, and may be useful for the development of drugs for Alzheimer's diseases.

Mutations in PRRT2 responsible for paroxysmal kinesigenic dyskinesias also cause benign familial infantile convulsions.

Paroxysmal kinesigenic dyskinesia (PKD (MIM128000)) is a neurological disorder characterized by recurrent attacks of involuntary movements. Benign familial infantile convulsion (BFIC) is also one of a neurological disorder characterized by clusters of epileptic seizures. The BFIC1 (MIM601764), BFIC2 (MIM605751) and BFIC4 (MIM612627) loci have been mapped to chromosome 19q, 16p and 1p, respectively, while BFIC3 (MIM607745) is caused by mutations in SCN2A on chromosome 2q24. Furthermore, patients with BFIC have been observed in a family concurrently with PKD. Both PKD and BFIC2 are heritable paroxysmal disorders and map to the same region on chromosome 16. Recently, the causative gene of PKD, the protein-rich transmembrane protein 2 (PRRT2), has been detected using whole-exome sequencing. We performed mutation analysis of PRRT2 by direct sequencing in 81 members of 17 families containing 15 PKD families and two BFIC families. Direct sequencing revealed that two mutations, c.649dupC and c.748C>T, were detected in all members of the PKD and BFIC families. Our results suggest that BFIC2 is caused by a truncated mutation that also causes PKD. Thus, PKD and BFIC2 are genetically identical and may cause convulsions and involuntary movements via a similar mechanism.

Correction: Sato-Akushichi et al. Choroidal Volume Evaluation after Photodynamic Therapy Using New Optical Coherence Tomography Imaging Algorithm. Pharmaceuticals 2021, 14, 1140.

Error in Figure [...].

One-Year Outcome of Combination Therapy with Full or Reduced Photodynamic Therapy and One Anti-Vascular Endothelial Growth Factor in Pachychoroid Neovasculopathy.

This paper evaluates a one-year treatment outcome after full or reduced photodynamic therapy (PDT) and anti-vascular endothelial growth factor (VEGF) combination therapy for pachychoroid neovasculopathy (PNV). After the initial combination therapy, a total of 29 eyes from 29 patients (16 for full treatment and 13 for reduced treatment), exhibited reduced, central retinal thickness and central choroidal thickness, and the improvements were maintained for 1 year after the initial combination therapy. Twenty-two eyes (75.9%) required no additional treatments for 1 year. The recurrence rate was 31.3% in the full treatment and 15.4% in the reduced treatment, with no significant differences between them. One shot of anti-VEGF and full or reduced PDT combination therapy had similar efficacy in treating PNV. Further prospective, large-scale, and long-term studies are required to determine a better treatment for PNV.

Fracture Resistance of Zirconia Abutments with or without a Titanium Base: An In Vitro Study for Tapered Conical Connection Implants.

Dental implants with tapered conical connections are often combined with zirconia abutments for esthetics; however, the effect of the titanium base on the implant components remains unclear. This study evaluated the effects of a titanium base on the fracture resistance of zirconia abutments and damage to the tapered conical connection implants. Zirconia (Z) and titanium base zirconia (ZT) abutments were fastened to Nobel Biocare (NB) implants and Straumann (ST) implants and subjected to static load testing according to ISO 14801:2016. The experiments were performed with 3 mm of the platform exposed (P3) and no platform exposed (P0). The fracture loads were statistically greater in the titanium base abutments than the zirconia abutments for the NB and ST specimens in the P0 condition. In the P3 condition of the ST specimens, the deformation volume of the ZT group was significantly greater than the Z group. The titanium base increased the fracture resistance of the zirconia abutments. Additionally, the titanium base caused more deformation in the P3 condition. The implant joint design may also affect the amount of damage to the implants when under a load. The mechanical properties of the abutment should be considered when selecting a clinical design.

Immunoglobulin G4-related disease accompanying a small intestinal ulcer: A case.

Immunoglobulin (Ig)G4-related disease (IgG4-RD) is a systemic condition associated with fibroinflammatory lesions and is characterized by elevated serum IgG4 levels and IgG4-positive cell infiltration into the affected tissues. It has been reported that IgG4-RD affects a variety of organs but uncommonly affects the gastrointestinal tract. In particular, there are few cases of lesions in the small intestine, except for sclerosing mesenteritis, which were mostly diagnosed from surgical specimens. Herein, we describe the case of a 70-year-old man who initially presented with abdominal pain, headache, later cognitive decline, and gait disturbance caused by IgG4-RD. Colonoscopy revealed irregular ulcers in the terminal ileum, and computed tomography of the head showed hypertrophic pachymeningitis. Numerous IgG4-positive cells were detected in the ileal and dural biopsies. We diagnosed the patient with IgG4-RD and started steroid pulse therapy. After initiation of treatment, the symptoms quickly improved. The patient was discharged from the hospital after starting oral prednisolone treatment (30 mg). The dosage was gradually reduced to 10 mg. A follow-up colonoscopy revealed scarring of the ileal ulcers. This case may provide valuable information regarding the endoscopic findings of small intestinal lesions in IgG4-RD.

Clinical improvement of membranous nephropathy after endoscopic resection of double early gastrointestinal cancers.

A-52-year-old man presented to our hospital with nephrotic syndrome caused by membranous nephropathy. Early gastric adenocarcinoma confined to the submucosa, and well-differentiated adenocarcinoma in a sigmoid adenoma were detected by screening endoscopy. Two years after complete endoscopic resection of these tumors, the estimated 24-h urinary protein excretion decreased, and serum total protein and albumin returned to their normal levels although he had no immunosuppressive therapy. Thus, this case was considered to be a case of secondary membranous nephropathy to cancer, although whether the pathogenesis was due to circulating or in situ immune complexes is unknown. The suspected antigen component of this immune complex can include carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9). In our patient, however, serum CEA and CA19-9 were within normal values during the clinical course, and detected cancers were early stage and very small. Recently, the existence of anti-mucin 1 (MUC1) antibodies before carcinogenesis and their usefulness for early detection of cancer were reported. We tried to stain tumors and glomeruli for MUC1 but, although we had positive findings in both tumors but not in glomeruli, the role of MUC1 in the pathogenesis of membranous nephropathy is unknown. To the best of our knowledge, paraneoplastic nephrosis caused by double early cancers expressing MUC1 in the gastrointestinal tract has not been previously reported.