Clinical features of double seronegative ocular myasthenia gravis.

PURPOSE: To clarify the clinical features of patients with Double seronegative (DS) ocular myasthenia gravis (OMG). METHODS: Sixty-one patients diagnosed with DS OMG at the Department of Ophthalmology, Hyogo Medical University Hospital over a 5-year period from 2017 were included. Patients were classified into three groups based on the initial examination findings: group P (ptosis alone), group M (ocular motility disorder alone), and group PM (combination of both). We retrospectively reviewed the patients and clarified their clinical features. RESULTS: There were 32 males and 29 females, with a mean age of 49.8 ± 20.9:1-82 years. Twenty-one patients (34.4%) were in group P, 23 (37.7%) in group M, and 17 (27.8%) in group PM. The proportion of males (73.9%) was significantly higher in group M compared with the other two groups. The diagnosis was proven by detection of neuromuscular junction (NMJ) disorder in 73.8%, oral pyridostigmine trial test in 13.1%, and eight patients (13.1%) in group M were diagnosed after surgical treatment. The clinical symptoms were resolved by oral pyridostigmine treatment in 54.1% of cases. CONCLUSION: About 30% of patients with DS OMG had no obvious NMJ disorder, and an oral pyridostigmine trial test was necessary to diagnose these patients. Although DS OMG is often considered as the mildest form of MG, its prognosis is not optimistic and it requires aggressive therapeutic intervention. TRIAL REGISTRATION: Trial registration number: 202104-750, "2016/4/18," retrospectively registered.

Yearly changes in cases of acute acquired comitant esotropia during a 12-year period.

PURPOSE: The number of patients with acute acquired comitant esotropia (AACE) has been increasing in Japan. The purpose of this study was to investigate the changes in the number and characteristics of patients with AACE examined in our institution during a 12-year period. METHODS: We retrospectively reviewed the medical records of patients with AACE aged < 30 years who suddenly developed diplopia or esotropia and were examined in Hyogo College of Medicine Hospital from January 2008 to December 2021. We investigated the association of the yearly changes in the number of patients with the age category, refractive error category, AACE type, esotropia type, and use or nonuse of smartphones. RESULTS: The total number of patients with AACE was 171, and this number significantly increased each year (Pearson correlation coefficient, 0.9450; p < 0.0001). Significant increases were found among students in junior high school and beyond, patients with myopia, patients with Bielschowsky type AACE, and patients with basic esotropia (p < 0.0001 for all). We compared two age groups, elementary school students and below versus junior high school students and above, and found that the rate of increase was significantly higher in the junior high school students and above (estimate, 1.951; p < 0.0001), and the non-myopia group and myopia group and found that the rate of increase was significantly higher in the myopia group (estimate, 1.891; p < 0.0001). Excessive use of smartphones was confirmed in 82 of 133 patients, and the rate of the increase in the number of patients with AACE was significantly greater among patients with than without excessive use of smartphones (estimate, 1.098; p = 0.0009). CONCLUSION: This study confirmed a significant increase in the number of patients with AACE in recent years. The excessive use of smartphones may be associated with the increase in AACE.

Identification of the roasting reaction products of ferulic and sinapic acids and their xanthine oxidase inhibitory activity.

The roasting reaction products of ferulic and sinapic acids were analyzed using high-performance liquid chromatography (HPLC) and their constituent compounds were isolated. Structural analysis revealed that the major compounds were oligomers of the corresponding 4-vinylphenols. The xanthine oxidase (XO) inhibitory activity of the isolated compounds was also measured. Moderate XO inhibitory activity of some oligomers afforded from ferulic acid was observed.

Structure and activity of the RNA-targeting Type III-B CRISPR-Cas complex of Thermus thermophilus.

The CRISPR-Cas system is a prokaryotic host defense system against genetic elements. The Type III-B CRISPR-Cas system of the bacterium Thermus thermophilus, the TtCmr complex, is composed of six different protein subunits (Cmr1-6) and one crRNA with a stoichiometry of Cmr112131445361:crRNA1. The TtCmr complex copurifies with crRNA species of 40 and 46 nt, originating from a distinct subset of CRISPR loci and spacers. The TtCmr complex cleaves the target RNA at multiple sites with 6 nt intervals via a 5' ruler mechanism. Electron microscopy revealed that the structure of TtCmr resembles a "sea worm" and is composed of a Cmr2-3 heterodimer "tail," a helical backbone of Cmr4 subunits capped by Cmr5 subunits, and a curled "head" containing Cmr1 and Cmr6. Despite having a backbone of only four Cmr4 subunits and being both longer and narrower, the overall architecture of TtCmr resembles that of Type I Cascade complexes.

Identification of a polyphenol and its antioxidant properties from the roasting reaction of alginic acid.

The radical scavenging activity of marine polysaccharides was enhanced by their high-temperature treatment (roasting reaction model). The product obtained from alginic acid exhibited maximum activity, and a radical scavenger, alginetin, was identified in the product. Its antioxidant activities were examined by chemical methods, which confirmed that it possessed a stoichiometrically greater antioxidant capacity than that of Trolox.

Novel Xanthine Oxidase (XO) inhibitory phenylindanes produced by thermal reaction of caffeic acid.

The products from the thermal reaction of chlorogenic and caffeic acids, which is a model process of roasting coffee beans, exhibited xanthine oxidase (XO) inhibitory activity. From caffeic acid, six inhibitory phenylindanes were identified, and a new phenylindane displayed the highest inhibitory activity among them. The activity of these phenylindanes may contribute to XO inhibition-related functions of roasted coffee beverages.

[The Significance of Genetic Testing for Acute Promyelocytic Leukemia].

The majority of patients with acute promyelocytic leukemia (APL) harbor the t (15;17) (q22;q12) transloca- tion, which results in the expression of PML-RARA mRNA. All-trans retinoic acid (ATRA) is a representa- tive molecular-targeted drug and is directed against PML-RARA. Therefore, the detection of PML-RARA mRNA has become indispensable for the diagnosis of APL and the decision regarding the treatment policy. Once the diagnosis is confirmed by genetic testing, ATRA-based induction therapy can be initiated. This is also applicable in atypical cases such as the M3 variant. Furthermore, after ATRA-based induction therapy, the curative effect is evaluated by quantitative PCR analysis. Thus, genetic testing is important in the follow-up of patients with APL. [Review].

[Discussion of Routine Laboratory Data of a Patient Complaining of Back Pain and General Fatigue].

Conventional reversed clinicopathological conference (RCPC) is an educational method to interpret labora- tory data. In this RCPC, physicians and several specialists in laboratory medicine discussed laboratory data of a patient with tuberculous spondylitis who complained of back pain and general fatigue. Then, they and the moderators held a question-and-answer session with an audience in a hall, and they tried to understand the detailed state of the patient. This discussion revealed the usefulness of RCPC to elucidate the clinical state of patient. At the same time, we can understand the limits of laboratory data analysis. [Review].

[Evaluation of Immature Platelet Fraction Measurements Using an Automated Hematology Analyzer, Sysmex XN].

The immature platelet fraction (IPF%) is a parameter for the automatic quantification of reticulated plate- lets and is considered to reflect platelet production. We evaluated IPF% measurements using an automated analyzer, the Sysmex XN. We measured the platelet counts and the IPF% values in 35 healthy subjects and 275 patients with various diseases using both the XN analyzer and a conventional analyzer, the Sysmex XE. Significant correlations in the platelet count and the IPF% were observed between the XN results and the XE results. In the same samples, significant inverse correlations were observed between the platelet count and the IPF% in both the XN and XE results. The coefficient of variation values for the platelet count and the IPF% measurements obtained using the XN analyzer were lower than those obtained using the XE ana- lyzer. In patients who had undergone hematopoietic stem cell transplantation, the IPF% measurements obtained using the XN analyzer increased several days before platelet recovery. IPF% measurements performed using the XN analyzer are adequate for clinical use. This parameter may be a useful marker for the prediction of platelet recovery. [Original].

[IgM-Lambda Type Paraproteinemia Interferes with Haptoglobin Measurements].

Paraproteinemia is a condition induced by an increase in paraprotein. Paraprotein is a monoclonal immu- noglobulin produced by plasma cells as a result of aging or malignancy. Paraprotein often induces a variety of laboratory test abnormalities by interfering with laboratory test reagents. The haptoglobin (Hp) measurement in a 55-year-old woman with IgM-lambda type paraproteinemia associated with lymphoplasmacytic lymphoma was found to be falsely low because of white-turbidity caused by an abnormal reaction. An in- creased absorbance was observed at 596 nm after the addition of the buffer reagent and was reduced by dilution with saline. This result indicates the existence of interfering substances in the patient's sample. To identify the inhibitors, we obtained the white-turbidity pellet by centrifugation of a mixture of the patient's serum and the Hp buffer reagent. A high IgM concentration was observed in the white-turbidity pellet. Moreover, a correlation was observed in a time series between the IgM concentration in sera and the extent of the turbidity during the Hp measurement. These results indicated that IgM was the main component of the white-turbidity. Next, we showed that the addition of the white-turbidity pellet to normal serum caused an increase in absorbance and a false low Hp value. A correlation was also observed in a time series be- tween the IgM concentration in sera and the rate of the Hp reduction. These results suggest that the false low Hp measurements were due to IgM present in the white-turbidity. In conclusion, false low values of Hp may occur in patients with IgM-lambda type paraproteinemia. Therefore, the presence or absence of an increase in absorbance after the addition of the buffer reagent in a time-course reaction may be required.

The shortest isoform of C/EBPß, liver inhibitory protein (LIP), collaborates with Evi1 to induce AML in a mouse BMT model.

Ecotropic viral integration site 1 (Evi1) is one of the master regulators in the development of acute myeloid leukemia (AML) and myelodysplastic syndrome. High expression of Evi1 is found in 10% of patients with AML and indicates a poor outcome. Several recent studies have indicated that Evi1 requires collaborative factors to induce AML. Therefore, the search for candidate factors that collaborate with Evi1 in leukemogenesis is one of the key issues in uncovering the mechanism of Evi1-related leukemia. Previously, we succeeded in making a mouse model of Evi1-related leukemia using a bone marrow transplantation (BMT) system. In the Evi1-induced leukemic cells, we identified frequent retroviral integrations near the CCAAT/enhancer-binding protein ß (C/EBPß) gene and overexpression of its protein. These findings imply that C/EBPß is a candidate gene that collaborates with Evi1 in leukemogenesis. Cotransduction of Evi1 and the shortest isoform of C/EBPß, liver inhibitory protein (LIP), induced AML with short latencies in a mouse BMT model. Overexpression of LIP alone also induced AML with longer latencies. However, excision of all 3 isoforms of C/EBPß (LAP*/LAP/LIP) did not inhibit the development of Evi1-induced leukemia. Therefore, isoform-specific intervention that targets LIP is required when we consider C/EBPß as a therapeutic target.

An oxidative coupling product of luteolin with cysteine ester and its enhanced inhibitory activity for xanthine oxidase.

Oxidative coupling reactions of several flavonoids with a cysteine ester (a radicalic and nucleophilic biochemical) were carried out and the abilities of the coupling products against xanthine oxidase (XO) were screened. One of the products, derived from luteolin, showed a notable inhibitory effect. A potent XO inhibitory compound was isolated from the complex mixture of the product of the coupling of luteolin and cysteine ethyl ester, and its structure was determined by NMR and MS analysis. The compound has a unique 1,4-thiazine ring unit on the luteolin B-ring and is inhibited XO 4.5 times more strongly than it did luteolin.

[Idiopathic dysplasia of undetermined/uncertain significance (IDUS) initially presenting erythroblastosis and hyperferritinemia].

A 64-year-old man was referred to our hospital because of mild anemia, erythroblasts in peripheral blood, and hyperferritinemia. In our hospital, no evidence of cytopenia was found. The peripheral blood smear showed pseudo-Pelger-Huët anomaly and giant platelets. The serum ferritin level was extremely high (1,405 ng/ml). A bone marrow examination was performed for further evaluation and revealed trilineage dysplasia. Since the patient did not fulfill the diagnostic criteria for cytopenias associated with myelodysplastic syndromes, he was diagnosed as having idiopathic dysplasia of undetermined/uncertain significance (IDUS). The SF3B1 mutation was identified in this patient, suggesting that he might be at a stage prior to myelodysplastic syndrome. Some IDUS patients reportedly progress to myeloid neoplasms, making careful observation essential. If morphological dysplasia in peripheral blood and hyperferritinemia are present, these findings suggest bone marrow failure syndromes. In such cases, further evaluation including a bone marrow examination may be required.

Lysyl 5-hydroxylation, a novel histone modification, by Jumonji domain containing 6 (JMJD6).

JMJD6 is reported to hydroxylate lysyl residues of a splicing factor, U2AF65. In this study, we found that JMJD6 hydroxylates histone lysyl residues. In vitro experiments showed that JMJD6 has a binding affinity to histone proteins and hydroxylates multiple lysyl residues of histone H3 and H4 tails. Using JMJD6 knock-out mouse embryos, we revealed that JMJD6 hydroxylates lysyl residues of histones H2A/H2B and H3/H4 in vivo by amino acid composition analysis. 5-Hydroxylysine was detected at the highest level in histones purified from murine testis, which expressed JMJD6 at a significantly high level among various tissues examined, and JMJD6 overexpression increased the amount of 5-hydroxylysine in histones in human embryonic kidney 293 cells. These results indicate that histones are additional substrates of JMJD6 in vivo. Because 5-hydroxylation of lysyl residues inhibited N-acetylation and N-methylation by an acetyltransferase and a methyltransferase, respectively, in vitro, histone 5-hydroxylation may have important roles in epigenetic regulation of gene transcription or chromosomal rearrangement.

Identification of crypto- and neochlorogenic lactones as potent xanthine oxidase inhibitors in roasted coffee beans.

Xanthine oxidase (XO) inhibitory activity has been found in boiling water extracts from roasted coffee beans. Therefore, assay-guided purification of the extracts was performed using size-exclusion column chromatography, and subsequently with reversed phase HPLC to afford lactone derivatives of chlorogenic acids. Among the tested lactones, crypto- and neochlorogenic lactones showed potent XO inhibitory activities compared with three major chlorogenic acids found in coffee beans. These XO inhibitory lactones may ameliorate gout and hyperuricemia in humans who drink coffee.

Antioxidant activities of cysteine derivatives against lipid oxidation in anhydrous media.

This study investigated antioxidant activities of cysteine derivatives of amino and carboxylic acid moieties against lipid oxidation in anhydrous acetonitrile. Only cysteine derivatives bearing free amino or carboxylate ion were found to exert potent antioxidant activities. Sequential proton loss and electron transfer-like proton shift and subsequent electron transfer (PS-ET) mechanism may facilitate the antioxidant activities of cysteine derivatives against lipid oxidation in anhydrous media.

Simplification of the genetic code: restricted diversity of genetically encoded amino acids.

At earlier stages in the evolution of the universal genetic code, fewer than 20 amino acids were considered to be used. Although this notion is supported by a wide range of data, the actual existence and function of the genetic codes with a limited set of canonical amino acids have not been addressed experimentally, in contrast to the successful development of the expanded codes. Here, we constructed artificial genetic codes involving a reduced alphabet. In one of the codes, a tRNAAla variant with the Trp anticodon reassigns alanine to an unassigned UGG codon in the Escherichia coli S30 cell-free translation system lacking tryptophan. We confirmed that the efficiency and accuracy of protein synthesis by this Trp-lacking code were comparable to those by the universal genetic code, by an amino acid composition analysis, green fluorescent protein fluorescence measurements and the crystal structure determination. We also showed that another code, in which UGU/UGC codons are assigned to Ser, synthesizes an active enzyme. This method will provide not only new insights into primordial genetic codes, but also an essential protein engineering tool for the assessment of the early stages of protein evolution and for the improvement of pharmaceuticals.

[The current status of genetic testing for leukemia].

The pathogenic chromosome translocations present in various hematological malignancies result in the formation of fusion genes, which are detected by a reverse transcription-polymerase chain reaction method. Furthermore, with this method, it is possible to sensitively detect minimal residual disease (MRD), which is difficult by a morphological testing. It has now been established that the detection of MRD is important for the diagnosis, treatment policy, evaluation of the prognosis, and monitoring of leukemia. In particular, quantitative analysis of MRD is important for evaluation of the curative effect and prediction of recurrence. In addition, mutation analysis is valuable to decide on the therapeutic protocol for imatinib-resistant patients, and the stratification of treatment for acute myeloid leukemia. At present, however, there is no standard laboratory procedure for genetic testing for leukemia. Here, the problems related to external precision management and analytical error are discussed.

[Difference in measurements of erythrocyte sedimentation rate obtained using two devices of the same model in a patient with marked leukocytosis].

The erythrocyte sedimentation rate (ESR) has been used as an index for inflammatory conditions, such as infectious diseases, autoimmune diseases, and malignancies. The ESR values of a 37-year-old male with marked leukocytosis due to chronic myeloid leukemia showed remarkable differences between two devices of the same model (Ves-Matic 30, DIESSE Diagnostica Senese). From the appearance of the tested tube after the ESR measurement, the values obtained using one device might have been falsely low, whereas the values obtained using the other device were likely to have been accurate. The difference of the ESR values between the two devices might have occurred by the false detection of transmitted light during the transition from the erythrocyte layer to the leukocyte layer. These findings suggest that in cases with marked leukocytosis the accuracy of ESR should be confirmed with the appearance of the test tube.

Localization of S100A2, S100A4, S100A6, S100A7, and S100P in the human hair follicle.

The hair follicle is a highly differentiated structure. In this study, we examined immunohistological localization of S100A2, S100A4, S100A6, S100A7, and S100P using specific monoclonal antibodies. S100A2 was strongly expressed in the entire outer-root sheath (ORS), but more weakly in cuticle and medulla in the bulb. S100A6, S100A7, and S100P were expressed in the innermost cells of ORS. The cuticular area was weakly positive for S100A2, S100A6, S100A7, and S100P. S100A4 was expressed in dendritic Langerhans cells and melanocytes. Sebaceous cells were variably immunopositive for S100A2, S100A6, and S100A7. A subset of dermal papilla cells expressed S100A4 and S100A6. None of the antibodies labeled the inner-root sheath. The distinct spatiostructural distributions of the S100 family proteins suggest that each protein is differentially involved in the physiological function of normal hair follicles.