Novel COL2A1 variants in Japanese patients with spondyloepiphyseal dysplasia congenita.

Spondyloepiphyseal dysplasia congenita (SEDC) is a multisystemic skeletal disorder caused by pathogenic variants in COL2A1. Here, we report the genotype-phenotype correlations in five Japanese patients with SEDC based on their clinical and radiological findings. All five patients had novel missense variants resulting in glycine substitutions (G474V, G543E, G567S, G594R, and G1170R). Genetic testing is important for early intervention for the extraskeletal complications of SEDC. Spondyloepiphyseal dysplasia congenita (SEDC) (OMIM#183900) is an autosomal dominant chondrodysplasia characterized by disproportionate short stature, abnormal epiphyses, flattened vertebral bodies (skeletal abnormalities), and extraskeletal features, including myopia, retinal degeneration with retinal detachment, and cleft palate. SEDC is caused by a heterozygous variant in the collagen II alpha 1 (COL2A1) gene.

Divergent variant patterns among 19 patients with Rubinstein-Taybi syndrome uncovered by comprehensive genetic analysis including whole genome sequencing.

Rubinstein-Taybi syndrome (RSTS) is characterized by dysmorphic facial features, broad thumbs, and intellectual disability. CREB-binding protein (CREBBP) or E1A-binding protein P300 (EP300) are causative genes. To elucidate the underlying genetic and genomic architecture related to the RSTS phenotype, we performed comprehensive genetic analysis targeting CREBBP and/or EP300 in 22 clinically diagnosed patients. During the 11-year study period, we used several analysis methods including high-resolution melting, array-based comparative genomic hybridization, panel-based exome sequencing, whole exome sequencing, and whole genome sequencing (WGS). We identified the causative variants in 19 patients (86.3%), but they were variable and complex, so we must combine multiple analysis methods. Notably, we found genetic alterations in the non-coding regions of two patients (10.5%, 2/19): scattered deletions including a partial 5'-untranslated region of CREBBP in one patient (all coding exons were intact), and a deep 229-bp intronic deletion in another patient, resulting in a splicing error. Furthermore, we identified rare clinical findings: two patients with an EP300 variant showed abnormal development of the neural tube, and one patient with a CREBBP variant had anorectal atresia with a cloaca. Our findings expand the allelic heterogeneity of RSTS, underscore the utility of comprehensive genetic analysis, and suggest that WGS may be a practical diagnostic strategy.

A novel method for isolating lymphatic endothelial cells from lymphatic malformations and detecting PIK3CA somatic mutation in these isolated cells.

PURPOSE: Tissue disaggregation and the cell sorting technique by surface markers has played an important role in isolating lymphatic endothelial cells (LECs) from lymphatic malformation (LM). However, this technique may have the drawback of impurities or result in isolation failure because it is dependent on surface marker expressions, the heterogeneity of which has been found in the lymphatic system. We developed a novel method for isolating LM-LECs without using whole tissue disaggregation. METHODS: Seven LM surgical specimens were collected from seven patients with LMs. LM-LECs were detached from the LM cyst wall by "lumen digestion" and irrigating the cystic cavity with trypsin, and maintained in culture. RESULTS: The cells formed a monolayer with a cobblestone-like appearance. Immunohistochemistry and quantitative RT-PCR of these cells revealed high expression of lymphatic-specific genes, confirming their identity as LM-LECs. The whole-exome sequencing and PIK3CA sequencing of these cells revealed somatic mutations in PIK3CA in all cases. CONCLUSIONS: We established a novel technique for isolating LM-LECs from LM tissue by "lumen digestion" without whole-tissue disaggregation. The limited incorporation of non-LM LECs in the isolate in our method could make it an important tool for investigating the heterogeneity of gene expression as well as mutations in LM-LECs.

Update of the genotype and phenotype of KMT2D and KDM6A by genetic screening of 100 patients with clinically suspected Kabuki syndrome.

Kabuki syndrome is characterized by a variable degree of intellectual disability, characteristic facial features, and complications in various organs. Many variants have been identified in two causative genes, that is, lysine methyltransferase 2D (KMT2D) and lysine demethylase 6A (KDM6A). In this study, we present the results of genetic screening of 100 patients with a suspected diagnosis of Kabuki syndrome in our center from July 2010 to June 2018. We identified 76 variants (43 novel) in KMT2D and 4 variants (3 novel) in KDM6A as pathogenic or likely pathogenic. Rare variants included a deep splicing variant (c.14000-8C>G) confirmed by RNA sequencing and an 18% mosaicism level for a KMT2D mutation. We also characterized a case with a blended phenotype consisting of Kabuki syndrome, osteogenesis imperfecta, and 16p13.11 microdeletion. We summarized the clinical phenotypes of 44 patients including a patient who developed cervical cancer of unknown origin at 16 years of age. This study presents important details of patients with Kabuki syndrome including rare clinical cases and expands our genetic understanding of this syndrome, which will help clinicians and researchers better manage and understand patients with Kabuki syndrome they may encounter.

Acute Effects of Whole-Body Vibration on Peripheral Blood Flow, Vibrotactile Perception and Balance in Older Adults.

BACKGROUND: Non-invasive application of whole-body vibration (WBV) has the potential for inducing improvements in impaired peripheral circulation, cutaneous sensation and balance among older adults. However, relevant studies have frequently applied high magnitudes of vibration and show conflicting and inconclusive results. Therefore, we attempted to ascertain the acute responses in those parameters from exposure of thirty older subjects to WBV of three different magnitudes, defined according to ISO 2631-1 (1997). METHODS: Each subject randomly underwent four sessions of intervention (three bouts of 1 min exposure with 1 min between-bout rests): WBV at 15, 20, or 25 Hz with a peak-to-peak displacement of 4 mm, or control condition. RESULTS: Both during and after intervention, dorsal foot skin blood flow increased significantly under 20 and 25 Hz exposure conditions with greater responses under the latter condition, the magnitude of which slightly exceeded the recommended value. Plantar vibrotactile perception showed significant increases after WBV exposure with overall greater responses under higher frequencies of vibration. In contrast, no WBV-induced change in balance was observed. CONCLUSIONS: WBV at 20 Hz with a magnitude within the recommended limit can be effective in inducing enhancements in peripheral blood flow; however, the same magnitude of vibration seems insufficient in improving balance among older adults.

An efficient genetic test flow for multiple congenital anomalies and intellectual disability.

BACKGROUND: Genetic testing has enabled the diagnosis of multiple congenital anomalies and/or intellectual disabilities. However, because of the phenotypic variability in these disorders, selection of an appropriate genetic test can be difficult and complex. For clinical examination, particularly in clinical facilities, a simple and standardized system is needed. METHODS: We compared microarray comparative genomic hybridization and clinical exome sequencing with regard to diagnostic yield, cost, and time required to reach a definitive diagnosis. After first performing G-banding for 200 patients with multiple congenital anomalies and/or intellectual disability, as a subsequent genetic test, microarray and clinical exome sequencing were compared with regard to diagnostic yield, cost, and time required. RESULTS: There was no obvious difference in the diagnostic rate between the two methods; however, clinical exome sequencing was superior in terms of cost and time. In addition, clinical exome sequencing could sufficiently identify copy number variants, and even smaller copy number variants could be identified. CONCLUSIONS: Clinical exome sequencing should be implemented earlier as a genetic test for undiagnosed patients with multiple congenital anomalies and/or intellectual disabilities. Our results can be used to establish inspection methods in clinical facilities.

A systematic review of studies investigating the effects of controlled whole-body vibration intervention on peripheral circulation.

The findings of the published studies investigating the changes in peripheral circulation induced by exposure to whole-body vibration (WBV) are not consensual or conclusive. Also, those studies did not consider the role of vibration magnitude on the peripheral circulatory responses making the interpretation of the observed findings difficult. We aimed to review the published literature investigating the effects of controlled WBV intervention on peripheral circulation by characterizing the relevant exposure conditions including vibration magnitudes, and ascertain the specific patterns of responses in peripheral circulation of the lower extremity from such exposure. A computerized search was performed in PubMed and Scopus using selected key search terms, and the relevant data were extracted. The vibration magnitude in the included studies frequently exceeded the limit specified in the International Standard ISO 2631-1 (1997) for such exposure. As observed, exposure to WBV with frequencies ≤30 Hz caused an improvement in peripheral blood flow (BF) and/or skin temperature (ST) of the lower extremity. In contrast, the responses in BF and ST from exposure to WBV of higher frequencies (31 Hz-50 Hz) produced conflicting results. The findings of this review indicate that WBV exposure with relatively lower frequency and magnitude can be safely and effectively used to induce improvements in peripheral circulation. For this purpose, the limits recommended by the ISO 2631-1 (1997) should be considered till safe and effective vibration-related parameters are established.

Clinical Evaluation of the Immunochromatographic System Using Silver Amplification for the Rapid Detection of Mycoplasma pneumoniae.

Mycoplasma pneumoniae infection is conventionally diagnosed using serum antibody testing, microbial culture, and genetic testing. Recently, immunochromatography-based rapid mycoplasma antigen test kits have been developed and commercialised for rapid diagnosis of M. pneumoniae infection. However, as these kits do not provide sufficient sensitivity and specificity, a rapid test kit with improved accuracy is desired. The present prospective study evaluated a rapid M. pneumoniae diagnostic system utilizing a newly developed silver amplification immunochromatography (SAI) system. We performed dilution sensitivity test and the prospective clinical study evaluating the SAI system. The subjects of the clinical study included both children and adults. All patients suspected to have mycoplasma pneumonia (169 patients) were sequentially enrolled. Twelve patients did not agree to participate and 157 patients were enrolled in the study. The results demonstrate excellent performance of this system with 90.4% sensitivity and 100.0% specificity compared with real-time polymerase chain reaction. When compared with loop-mediated isothermal amplification (LAMP) methods, the results also demonstrate a high performance of this system with 93.0% sensitivity and 100.0% specificity. The SAI system uses a dedicated device for automatic analysis and reading, making it highly objective, and requires less human power, supporting its usefulness in clinical settings.

Novel SYNGAP1 variant in a patient with intellectual disability and distinctive dysmorphisms.

We describe a novel de novo heterozygous variant in SYNGAP1 (c.1741C>T, p.R581W), identified through targeted resequencing in an 8-year-old boy with intellectual disability, autism spectrum disorder, distinctive dysmorphic features, and no seizures. Our data strongly suggest that the SYNGAP1 variant is causative of intellectual disability in this patient.

Clinical significance of serum complement factor 3 in patients with type 2 diabetes mellitus.

AIMS: Although serum complement factor 3 (C3) is an acute phase reactant mainly synthesized in the liver, several recent studies have shown high C3 gene expression in adipose tissue (AT). However, the relationship between C3 and AT levels has not been fully clarified in type 2 diabetes mellitus (T2DM) patients. METHODS: A total of 164 T2DM patients (109men and 55 women) participated in this cross-sectional study. A computed tomography scan was performed to measure visceral, subcutaneous, and total AT. The correlation between these factors and C3 levels was examined using Pearson's correlation analysis. A multivariate regression model was used to assess an independent determinant associated with C3 levels after adjusting the explanatory variables (i.e., all ATs [visceral, subcutaneous, and total], and clinical features [sex, age, body mass index, waist circumference, glycated hemoglobin, duration of diabetes, systolic blood pressure, diastolic blood pressure, aspartate aminotransferase levels, alanine aminotransferase levels, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, log(triglyceride levels), estimated glomerular filtration rate, and log(high-sensitivity C-reactive protein levels)]). RESULTS: Serum C3 levels were correlated with visceral, subcutaneous, and total AT among both men (r=0.505, p<0.001; r=0.545, p<0.001; r=0.617, p<0.001, respectively) and women (r=0.396, p=0.003; r=0.517, p<0.001; r=0.548, p<0.001, respectively). In the multivariate regression model, the association between total AT and C3 levels remained significantly positive (ß=0.490, p<0.001). CONCLUSIONS: Serum C3 levels are associated with visceral, subcutaneous, and total AT in T2DM patients. Furthermore, C3 levels seem to be a marker for overall adiposity rather than regional adiposity.

A systematic review of diagnostic performance of quantitative tests to assess musculoskeletal disorders in hand-arm vibration syndrome.

The purpose was to systematically review the published reports for the clinical utility of quantitative objective tests commonly used for diagnosing musculoskeletal disorders in hand-arm vibration syndrome (HAVS). Two reviewers independently conducted a computerized literature search in PubMed and Scopus using predefined criteria, and relevant papers were identified. The articles were screened in several stages and considered for final inclusion. Quality of the selected papers was evaluated by a modified QUADAS tool. Relevant data were extracted as necessary. For this review, only 4 relevant studies could be identified for detailed examination. Grip strength, pinch strength, and Purdue pegboard tests were commonly used with their reported sensitivity and specificity ranging between 1.7 to 65.7% and 65.2 to 100%, 1.7 to 40% and 94 to 100%, and 44.8 to 85% and 78 to 95%, respectively. A considerable difference across the studies was observed with respect to patient and control populations, diagnostic performance and cut-off values of different tests. Overall, currently available English-language limited literature do not provide enough evidence in favour of the application of grip strength and pinch strength tests for diagnosing musculoskeletal injuries in HAVS; Purdue pegboard test seems to have some diagnostic value in evaluating impaired dexterity in HAVS.

Running suture versus interrupted suture for vesicourethral anastomosis in retropubic radical prostatectomy: a randomized study.

OBJECTIVES: To determine the feasibility of vesicourethral anastomosis using running suture during retropubic radical prostatectomy and to compare the surgical outcomes of vesicourethral anastomosis using running suture with those of the standard interrupted suture technique. METHODS: A total of 60 patients undergoing radical prostatectomy from 2010 to 2012 at the Yamaguchi University Hospital, Japan were included in the present study, and were randomly assigned to vesicourethral anastomosis using running suture (n = 30 patients) or a standard interrupted suture technique group (n = 30 patients). Vesicourethral anastomosis using running suture was carried out with 12-bite sutures using 3-0 poliglecaprone. The primary end-point was the time to catheter removal. Patients' health-related quality of life was assessed using the Expanded Prostate Cancer Index Composite in 56 patients (28 patients in each group). RESULTS: No significant difference was found in the median suturing time between the two study groups (both 19 min, P = 0.449). The time to catheter removal was significantly better in the vesicourethral anastomosis using running suture group (hazard ratio 5.23, 95% confidence interval 1.73-17.65, P = 0.003). The pad-free rate was significantly higher in the vesicourethral anastomosis using running suture group at 1 month after surgery (20.7% vs 3.3%, P = 0.0463); however, there was no significant difference at 3 months and beyond. The Expanded Prostate Cancer Index Composite urinary and bowel summary scores at 1 month were significantly better in the vesicourethral anastomosis using running suture patients (both P < 0.01), though no significant difference was observed thereafter. A vesicourethral anastomosis stricture was noted in three patients (10%) in the standard interrupted suture technique group, and none in the vesicourethral anastomosis using running suture group. CONCLUSION: Running suture for vesicourethral anastomosis is feasible during retropubic radical prostatectomy. Furthermore, it offers better outcomes than the conventional standard interrupted suture technique, with a higher likelihood of improvement in patients' health-related quality of life.

Analysis of long-term stored plasma samples for investigation into the pathophysiology of vibration-induced white finger: preliminary results.

Blood samples were collected 25 yr ago from hand-arm vibration syndrome patients with vibration-induced white finge/VWF (VWF+ group) and without it (VWF- group), and healthy controls (n=12 in each group), and stored at -80 °C. The subjects provided venous blood twice: at baseline, and after cold exposure at 7 °C for 25 min. Blood specimens were analyzed for plasma endothelin-1 (ET-1) by an enzyme-linked immunosorbent assay. Baseline concentration of plasma ET-1 was found to be significantly larger in the VWF- group than the control group, whereas no such difference was observed for the VWF+ group. However, the %change was larger in the VWF+ group (107.73 ± 30.49%) than the other two groups, and more subjects in the VWF+ group showed the maximum increase in ET-1 than the other two groups. In conclusion, ET-1 appears to have a role in the pathophysiology of VWF.

[Relatedness of occupational exposure to whole-body vibration and health, principally back symptoms].

OBJECTIVES: DISCUSSION is continuing about whether or not to update the Occupational Exposure Limit for Whole Body Vibration exposure (OELWBV) which was established in 1975 as a part of the Recommendation of Occupational Exposure Limits (OEL), issued by the Japan Society for Occupational Health. The objectives of this study were to clarify the necessity of the update and the adverse health effects of whole-body vibration by reviewing the literature and to help to compile a for detailed revisions. SUBJECTS AND METHODS: Mainly referring to our preceding review, the current OELWBV exposure was examined. The relatedness of adverse health effects, principally back symptoms and occupational exposure to whole-body vibration (WBV) was examined by reviewing review papers, the original English papers referred to in those papers, and original English and Japanese references from 2002 to 2010 retrieved through a MEDLINE search. RESULTS: The results show the necessity of updating the present OELWBV as soon as possible. The overwhelming majority of the literature consists of epidemiological studies showing the relatedness of WBV to back symptoms. Thirty epidemiological studies out of 28 literatures clearly show the presence of a relationship between WBV and back symptoms. Various indices of exposure to WBV and back symptoms show the dose response relationships. Among these, a few studies used A(sum)(8) (equivalent 8-hour r.m.s. acceleration value for the combined frequency-weighted vibration of three diagonal, that is x, y and z, axes) as the exposure index. All of these studies reported some dose-response relationship between WBV exposure and back symptoms. DISCUSSION: Despite the clear presence or absence of a relationship between WBV and back symptoms, 0.5 m/s(2) seems to be the unconfirmed threshold of risk. Focusing on data of A(sum)(8), we suggest that the risk increases over the magnitude of 0.30 m/s(2). To clarify this, further epidemiological studies and others should focus on A(sum)(8) around 0.30 m/s(2). CONCLUSIONS: The present OEL of WBV should be examined from the viewpoint of the risks of back symptoms referring to the epidemiological studies that show the larger risks of back symptoms in comparison with unexposed referent workers. We conclud that when updating the present OELWBV, A(sum)(8) should be taken into account for back symptoms, referring to the findings of epidemiological studies.

Review of different quantification methods for the diagnosis of digital vascular abnormalities in hand-arm vibration syndrome.

OBJECTIVE: This study was undertaken to review the diagnostic ability of different quantification methods in the assessment of vibration-induced white finger (VWF), the typical clinical manifestation of vascular injuries in Hand-arm Vibration Syndrome (HAVS). METHODS: A literature search of original and major review articles related to the quantification techniques for diagnosing vascular injuries in HAVS was performed. Relevant data from the publications were extracted and included in this study for reporting and discussion. RESULTS: Few studies were available to substantiate the diagnostic techniques using the nail press test and nailfold capillaroscopy. Also, few studies were found to conclusively demonstrate the diagnostic ability using thermometry and thermography incorporated with cold provocation. Some recent reports raised the question regarding the diagnostic ability of finger plethysmography; but by virtue of its comparable assessment parameters and better diagnostic performance, plethysmography appears to be a suitable diagnostic method. In noninvasive quantification of vascular injuries, diagnostic techniques like laser Doppler perfusion imaging and nailfold capillaroscopy require further evaluation in future studies. CONCLUSIONS: For a reliable objective diagnosis of VWF at present, quantification of vascular responses using a test battery including established methods like thermometry or thermography and strain gauge plethysmography appears to be useful.

Diagnostic performance of cold provocation test with hands immersion in water at 10°C for 5 min evaluated in vibration-induced white finger patients and matched controls.

PURPOSE: This study aimed to explore the diagnostic ability of the cold provocation test with hands immersion in water at 10°C for 5 min in diagnosing vibration-induced white finger (VWF). METHODS: Finger skin temperature (FST) was measured in 20 VWF patients and 20 matched healthy controls, at palmar side of the distal phalanges of fingers from both hands before, during, and after hands immersion in water at 10°C (for 5 min with waterproof coverings put on both hands). Data from 4 fingers (except thumb) were evaluated at five time points: just before immersion, last minute during immersion, and at 5th, 10th, and 15th min during the post-immersion or recovery period. RESULTS: A positive group difference between patients and controls was revealed during the recovery period. During recovery at 95 and 70% specificity, the sensitivity ranged from 20 to 30% and 50 to 70% for evaluation with average FST for 4 fingers and 15-35% and 60-65% for evaluation with minimum FST among 4 fingers, respectively. Overall, evaluation of absolute FST at 15th min of recovery offered better diagnostic ability. CONCLUSIONS: The cold provocation test with hands immersion in water at 10°C for 5 min could discriminate VWF patients from healthy controls; however, this test has a limited diagnostic value in diagnosing patients with VWF.

Vibrotactile perception at glabrous and nonglabrous skin of fingers: repeatability of measurements and changes induced by acute vibration exposure.

OBJECTIVES: We sought to determine the within-session and between-session repeatability of vibrotactile perception threshold (VPT) measurements and the response patterns in VPT induced by acute exposure to short-term vibration from grasping a vibrating handle, at both glabrous and nonglabrous skin of fingers. METHODS: Baseline VPT was recorded twice at glabrous and nonglabrous side of fingers in the right hand of eight healthy volunteers. Then, the subjects were exposed to three exposure conditions (vibration at 31.5 Hz and 250 Hz, and no vibration), from gripping a vertical handle by the right hand, conducted on 3 different days at an interval of 1-3 wk. After exposure, the subjects released the hand and further VPT measurements at each location were made. RESULTS: Compared to the nonglabrous side, VPT measurements at the glabrous side demonstrated better within-session and between-session repeatability with lower coefficient of repeatability and higher intraclass correlation coefficient. After exposure, a significant increase in VPT was noted under both 31.5 Hz and 250 Hz (p<0.05-0.001) exposure conditions in the glabrous finger. In the nonglabrous finger, a pronounced increase in VPT was revealed under 250 Hz exposure condition (p=0.05). CONCLUSIONS: While measuring VPT at glabrous and/or nonglabrous fingers, the importance of the site of measurement should be considered; the repeatability for such measurements appears to be better at the glabrous site. At high frequency, vibrotactile perception appears to be affected in both glabrous and nonglabrous skin from acute vibration exposure.

Quality of life of arsenicosis patients in an arsenic-affected rural area in Bangladesh.

The objective of this cross-sectional comparative study was to assess quality of life (QOL) of arsenicosis patients. A total of 104 arsenicosis patients, 104 arsenic-exposed individuals without arsenicosis (nonpatients), and 104 individuals not exposed to arsenic (controls) were interviewed based on a questionnaire constructed based on the WHOQOL-BREF instrument. The facets of life considered for assessing QOL were physical health domain, psychological domain, social relationships domain, and environment domain. All the domain-wise QOL scores of the arsenicosis patients were significantly smaller than those of the controls as well as of the nonpatients. The findings of the study assessing QOL of arsenicosis patients might be helpful to understand the condition and justify for the policy makers to adopt adequate measures to alleviate the sufferings through prevention, treatment, and rehabilitation.