Abnormality of vascular elastic fibers in the macular mouse and a patient with Menkes'' disease: ultrastructural and immunohistochemical study

The macular mouse is a mutant mouse with the same gene abnormality as that of Menkes' disease, and it exhibits symptoms and abnormalities similar to those of Menkes' disease. In an electron microscopic study, we examined morphological changes in the internal elastic lamina (IEL) of the elastic arteries (EA) and the muscular arteries (MA) in a patient with Menkes' disease and in the macular mouse, an animal model of this disease. The IEL of the EA was significantly thinner in the macular mouse than that in controls, but the IEL of the MA in the macular mouse was significantly thicker than that of the controls. These contrary results for the thickness of the IEL in the MA and the EA in this animal model of Menkes' disease may reflect differences in the anatomical and pathophysiological properties of the two types of vessels.     

Diagnostic strategy for inherited hypomagnesemia

Background

Hereditary hypomagnesemia is difficult to diagnose accurately because of its rarity and the variety of causative genes. We established a flowchart for identifying responsible genes for hypomagnesemia, and we confirmed its diagnostic efficacy in patients with suspected inherited hypomagnesemia.

Methods

We established a flowchart and applied it to five index cases with suspected inherited hypomagnesemia. Direct sequence analysis was used to detect the causative gene variants in four cases, and targeted sequencing analysis using next-generation sequencing (NGS) of all causative genes for hypomagnesemia was used in one.

Results

Expected pathogenic variants were detected in the HNF1B, TRPM6, CLDN16, CASR, or SLC12A3 gene in all five cases. The results of all genetic analyses were consistent with the clinical diagnostic results using the flowchart.

Conclusions

Accurate genetic diagnosis is crucial for estimating the prognosis, detecting complications in organs other than the kidneys, and for directing genetic counseling. The developed flowchart for identifying responsible genes for hypomagnesemia was useful for diagnosing inherited hypomagnesemia. In addition, NGS analysis will help to resolve clinical difficulties in making an accurate diagnosis and thus improve the diagnostic strategy for inherited hypomagnesemia.

     

Responsiveness of the Self-Administered Foot Evaluation Questionnaire (SAFE-Q) in patients with hallux valgus

Background

In this study, we investigated the responsiveness of the Self-Administered Foot Evaluation Questionnaire (SAFE-Q) for patient's assessment before and after hallux valgus surgery.

Correlation between parametric imaging using contrast ultrasound and the histological differentiation of hepatocellular carcinoma.

Aim: To determine whether parametric imaging correlates with the degree of histological differentiation of hepatocellular carcinoma (HCC). Methods: The samples comprised 49 nodules diagnosed histologically as HCC: 19 well differentiated (w-HCC), 22 moderately differentiated (m-HCC), and eight poorly differentiated (p-HCC). The ultrasound (US) equipment used was SSA-770 A (Toshiba Medical Systems, Otawara, Japan) and the contrast agent was SonoVue (Bracco, Milan, Italy). After 1.5 mL of SonoVue was injected intravenously and staining of the tumors and parenchyma was confirmed, microbubbles in the scanned volume were eliminated using high mechanical index (MI) scanning frames. The "arrival time (T(A)) images," reflecting beta-values, were displayed with color codes at the phase after reperfusion. Images at the phase when the staining reached a plateau (90-180 s) were used as "A images," reflecting A values. These images were compared between each histological grade of differentiation. Results: Analysis of T(A) images indicated that beta-values in m-HCC were higher than those in the adjacent non-tumor parenchyma in all 22 samples and also were significantly higher than in the other HCCs (P < 0.001 for w-HCC; P < 0.05 for p-HCC). Furthermore, beta-values in p-HCC samples had significantly larger variations in terms of time and space than in the other HCCs (P < 0.001 for w-HCC; P < 0.01 for m-HCC). Analysis of A images indicated that the A value for w-HCC was significantly higher than those for either m-HCC or p-HCC (P < 0.001). Conclusion: Both T(A) and A images were useful for diagnosing the histological differentiation of HCC.     

Association of WT1 IgG antibody against WT1 peptide with prolonged survival in glioblastoma multiforme patients vaccinated with WT1 peptide

We previously evaluated Wilms’ tumor gene 1 (WT1) peptide vaccination in a large number of patients with leukemia or solid tumors and have reported that HLA‐A*24:02 restricted, 9‐mer WT1‐235 peptide (CYTWNQMNL) vaccine induces cellular immune responses and elicits WT1‐235‐specific cytotoxic T lymphocytes (CTLs). However, whether this vaccine induces humoral immune responses to produce WT1 antibody remains unknown. Thus, we measured IgG antibody levels against the WT1‐235 peptide (WT1‐235 IgG antibody) in patients with glioblastoma multiforme (GBM) receiving the WT1 peptide vaccine. The WT1‐235 IgG antibody, which was undetectable before vaccination, became detectable in 30 (50.8%) of a total of 59 patients during 3 months of WT1 peptide vaccination. The dominant WT1‐235 IgG antibody subclass was Th1‐type, IgG₁ and IgG₃. WT1‐235 IgG antibody production was significantly and positively correlated with both progression‐free survival (PFS) and overall survival (OS). Importantly, the combination of WT1‐235 IgG antibody production and positive delayed type‐hypersensitivity (DTH) to the WT1‐235 peptide was a better prognostic marker for long‐term OS than either parameter alone. These results suggested that WT1‐235 peptide vaccination induces not only WT1‐235‐specific CTLs as previously described but also WT1‐235‐specific humoral immune responses associated with antitumor cellular immune response. Our results indicate that the WT1 IgG antibody against the WT1 peptide may be a useful predictive marker, with better predictive performance in combination with DTH to WT1 peptide, and provide a new insight into the antitumor immune response induction in WT1 peptide vaccine‐treated patients.     

Stability for long-term storage and reproducibility of positivity in the panel test slide prepared with the polyacrylamide-based artificial sputum

OBJECTIVE: A novel artificial sputum has been developed using polyacrylamide, cultured THP-1 cell and BCG-Pasteur. Smears prepared with this artificial sputum are similar to actual sputum and has feasibility to set any positivity grades. Long-term storage and reproducibility of the positivity was examined to support further availability. METHOD: The artificial sputa were stored for up to 9 months at room temperature, 4 degrees C and -20 degrees C. Then, smears were prepared and their macroscopic and microscopic appearance were examined compared with smears from freshly prepared artificial sputum. Furthermore, smears with different positivities (+/-, 1 +, 2 + and 3 +) were prepared and examined by several trained technicians, and the reproducibility of the original sputum positivity was determined. RESULTS: Macroscopic and microscopic appearance of smears prepared from long-term stored artificial sputum showed little changes compared with smears of freshly prepared artificial sputum. The positivity of these smears fell in their original grade. A total of 91 smears were prepared from artificial sputum with different positivity and examined by trained technicians. Although 3 out of 36 +/- smears were determined as negative, all of the remaining smears were evaluated correctly. DISCUSSION: This study confirmed that the artificial sputum and the smears have long-term storage stability and reproducibility in the positivity. These results suggest that the artificial sputum can be widely used to perform external quality assessment in many countries, including high prevalence countries.