Genomic Organization of the Human Arpp Gene

A novel ankyrin-repeated protein, Arpp, is specifically expressed in skeletal and cardiac muscles. Arpp protein is homologous, in its amino acid sequences (52.7% identity), to Carp protein which is proposed to be a putative genetic marker for cardiac hypertrophy. In this study, we isolated the human Arpp gene by screening a human genomic library and analyzed the genomic structure and its 5' flanking region. The Arpp gene was found to encompass a sequence of 11 kb and to consist of 9 exons. The translational initiation site and the stop codon were found to be located at exon 1 and exon 9, respectively. Each exon from 5 to 8 was found to encode 1 of the 4 ankyrin-like domains, respectively. The 2.7 kb upstream of exon 1 was sequenced. The TATA box was identified 29 bp upstream of the transcriptional start site, and multiple putative regulatory elements including the E box and upstream stimulating factor-1 were distributed within the proximal promoter regions. Since these elements were also found in the promoter region of the mouse Arpp gene, they may play an important role in the transcriptional regulation of both human and murine Arpp genes

Mechanisms of tissue degeneration mediated by periostin in spinal degenerative diseases and their implications for pathology and diagnosis: a review

Periostin (POSTN) serves a dual role as both a matricellular protein and an extracellular matrix (ECM) protein and is widely expressed in various tissues and cells. As an ECM protein, POSTN binds to integrin receptors, transduces signals to cells, enabling cell activation. POSTN has been linked with various diseases, including atopic dermatitis, asthma, and the progression of multiple cancers. Recently, its association with orthopedic diseases, such as osteoporosis, osteoarthritis resulting from cartilage destruction, degenerative diseases of the intervertebral disks, and ligament degenerative diseases, has also become apparent. Furthermore, POSTN has been shown to be a valuable biomarker for understanding the pathophysiology of orthopedic diseases. In addition to serum POSTN, synovial fluid POSTN in joints has been reported to be useful as a biomarker. Risk factors for spinal degenerative diseases include aging, mechanical stress, trauma, genetic predisposition, obesity, and metabolic syndrome, but the cause of spinal degenerative diseases (SDDs) remains unclear. Studies on the pathophysiological effects of POSTN may significantly contribute toward the diagnosis and treatment of spinal degenerative diseases. Therefore, in this review, we aim to examine the mechanisms of tissue degeneration caused by mechanical and inflammatory stresses in the bones, cartilage, intervertebral disks, and ligaments, which are crucial components of the spine, with a focus on POSTN

Downregulation of SAV1 plays a role in pathogenesis of high-grade clear cell renal cell carcinoma

<p>Abstract</p> <p>Background</p> <p>Clinical outcome of patients with high-grade ccRCC (clear cell renal cell carcinoma) remains still poor despite recent advances in treatment strategies. Molecular mechanism of pathogenesis in developing high-grade ccRCC must be clarified. In the present study, we found that SAV1 was significantly downregulated with copy number loss in high-grade ccRCCs. Therefore, we investigated the SAV1 function on cell proliferation and apoptosis in vitro. Furthermore, we attempted to clarify the downstream signaling which is regulated by SAV1.</p> <p>Methods</p> <p>We performed array CGH and gene expression analysis of 8 RCC cell lines (786-O, 769-P, KMRC-1, KMRC-2, KMRC-3, KMRC-20, TUHR4TKB, and Caki-2), and expression level of mRNA was confirmed by quantitative RT-PCR (qRT-PCR) analysis. We next re-expressed SAV1 in 786-O cells, and analyzed its colony-forming activity. Then, we transfected siRNAs of SAV1 into the kidney epithelial cell line HK2 and renal proximal tubule epithelial cells (RPTECs), and analyzed their proliferation and apoptosis. Furthermore, the activity of YAP1, which is a downstream molecule of SAV1, was evaluated by western blot analysis, reporter assay and immunohistochemical analysis.</p> <p>Results</p> <p>We found that SAV1, a component of the Hippo pathway, is frequently downregulated in high-grade ccRCC. SAV1 is located on chromosome 14q22.1, where copy number loss had been observed in 7 of 12 high-grade ccRCCs in our previous study, suggesting that gene copy number loss is responsible for the downregulation of SAV1. Colony-forming activity by 786-O cells, which show homozygous loss of SAV1, was significantly reduced when SAV1 was re-introduced exogenously. Knockdown of SAV1 promoted proliferation of HK2 and RPTEC. Although the phosphorylation level of YAP1 was low in 786-O cells, it was elevated in SAV1-transduced 786-O cells. Furthermore, the transcriptional activity of the YAP1 and TEAD3 complex was inhibited in SAV1-transduced 786-O cells. Immunohistochemistry frequently demonstrated nuclear localization of YAP1 in ccRCC cases with SAV1 downregulation, and it was preferentially detected in high-grade ccRCC.</p> <p>Conclusions</p> <p>Taken together, downregulation of SAV1 and the consequent YAP1 activation are involved in the pathogenesis of high-grade ccRCC. It is an attractive hypothesis that Hippo signaling could be candidates for new therapeutic target.</p

Helicobacter pylori infection and gastroduodenal diseases in Vietnam: a cross-sectional, hospital-based study

<p>Abstract</p> <p>Background</p> <p>The rate of <it>H. pylori </it>infection in Vietnam is reportedly high, but the spectrum of <it>H. pylori</it>-associated gastroduodenal diseases has not been systematically investigated. Moreover, despite the similarities of ethnicity and diet, the age-standardized incidence rate of gastric cancer in the northern city of Hanoi is higher than that in the southern city of Ho Chi Minh, but the reason for this phenomenon is unknown. The virulence of Vietnamese <it>H. pylori </it>has also not been investigated in detail.</p> <p>Methods</p> <p>Individuals undergoing esophagogastroduodenoscopy were randomly recruited. <it>H. pylori </it>infection status was determined based on the combined results of culture, histology, immunohistochemistry, rapid urine test and serum ELISA. Peptic ulcer (PU) and gastroesophageal reflux disease was diagnosed by endoscopy, and chronic gastritis was determined histologically. <it>H. pylori </it>virulence factors were investigated by PCR and sequencing.</p> <p>Results</p> <p>Among the examined patients, 65.6% were infected with <it>H. pylori</it>. The prevalence of infection was significantly higher in those over 40 years of age than in those aged ≤40. Chronic gastritis was present in all <it>H. pylori</it>-infected individuals, 83.1% of whom had active gastritis, and 85.3% and 14.7% had atrophy and intestinal metaplasia, respectively. PU was present in 21% of infected patients, whereas its incidence was very low in non-infected individuals. The prevalence of PU was significantly higher in Hanoi than in Ho Chi Minh. The prevalence of <it>vacA m1</it>, which has been identified as an independent risk factor for PU in Vietnam, was significantly higher among <it>H. pylori </it>isolates from Hanoi than among those from Ho Chi Minh.</p> <p>Conclusions</p> <p><it>H. pylori </it>infection is common in Vietnam and is strongly associated with PU, active gastritis, atrophy and intestinal metaplasia. <it>vacA m1 </it>is associated with an increased risk for PU and might contribute to the difference in the prevalence of PU and gastric cancer between Hanoi and Ho Chi Minh.</p

Genomic Profiling of Submucosal-Invasive Gastric Cancer by Array-Based Comparative Genomic Hybridization

Genomic copy number aberrations (CNAs) in gastric cancer have already been extensively characterized by array comparative genomic hybridization (array CGH) analysis. However, involvement of genomic CNAs in the process of submucosal invasion and lymph node metastasis in early gastric cancer is still poorly understood. In this study, to address this issue, we collected a total of 59 tumor samples from 27 patients with submucosal-invasive gastric cancers (SMGC), analyzed their genomic profiles by array CGH, and compared them between paired samples of mucosal (MU) and submucosal (SM) invasion (23 pairs), and SM invasion and lymph node (LN) metastasis (9 pairs). Initially, we hypothesized that acquisition of specific CNA(s) is important for these processes. However, we observed no significant difference in the number of genomic CNAs between paired MU and SM, and between paired SM and LN. Furthermore, we were unable to find any CNAs specifically associated with SM invasion or LN metastasis. Among the 23 cases analyzed, 15 had some similar pattern of genomic profiling between SM and MU. Interestingly, 13 of the 15 cases also showed some differences in genomic profiles. These results suggest that the majority of SMGCs are composed of heterogeneous subpopulations derived from the same clonal origin. Comparison of genomic CNAs between SMGCs with and without LN metastasis revealed that gain of 11q13, 11q14, 11q22, 14q32 and amplification of 17q21 were more frequent in metastatic SMGCs, suggesting that these CNAs are related to LN metastasis of early gastric cancer. In conclusion, our data suggest that generation of genetically distinct subclones, rather than acquisition of specific CNA at MU, is integral to the process of submucosal invasion, and that subclones that acquire gain of 11q13, 11q14, 11q22, 14q32 or amplification of 17q21 are likely to become metastatic

Upregulation of miR-30d in Persistent AF

Background: Atrial fibrillation (AF) begets AF in part due to atrial remodeling, the molecular mechanisms of which have not been completely elucidated. This study was conducted to identify microRNA(s) responsible for electrical remodeling in AF. Methods and Results: The expression profiles of 1205 microRNAs, in cardiomyocytes from patients with persistent AF and from age-, gender-, and cardiac function-matched control patients with normal sinus rhythm, were examined by use of a microRNA microarray platform. Thirty-nine microRNAs differentially expressed in AF patients’ atria were identified, including miR-30d, as a candidate responsible for ion channel remodeling by in silico analysis. MiR-30d was significantly upregulated in cardiomyocytes from AF patients, whereas the mRNA and protein levels of CACNA1C/ Cav1.2 and KCNJ3/Kir3.1, postulated targets of miR-30d, were markedly reduced. KCNJ3/Kir3.1 expression was downregulated by transfection of the miR-30 precursor, concomitant with a reduction of the acetylcholine-sensitive inward-rectifier K+ current (IK.ACh). KCNJ3/Kir3.1 (but not CACNA1C/Cav1.2) expression was enhanced by the knockdown of miR-30d. The Ca2+ ionophore, A23187, induced a dose-dependent upregulation of miR-30d, followed by the suppression of KCNJ3 mRNA expression. Blockade of protein kinase C signaling blunted the [Ca2+]i-dependent downregulation of Kir3.1 via miR-30d. Conclusions: The downward remodeling of IK.ACh is attributed, at least in part, to deranged Ca2+ handling, leading to the upregulation of miR-30d in human AF, revealing a novel post-transcriptional regulation of IK.ACh

Minimally Invasive Endoscopic Approach to the Cervicothoracic Junction for Vertebral Osteomyelitis

The selection of an anterior, lateral, or posterior approach to the cervicothoracic junction for surgical treatment of vertebral osteomyelitis is still a matter of debate. These ordinary approaches generally require an extensile exposure. This article describes a less invasive approach case of a vertebral osteomyelitis of T2/3 using a video-assisted operating technique of thoracic surgery (VATS). A 78-year-old female underwent anterior debridement and interbody fusion with bone graft at T2/3 using a lateral surgical approach through a right thoracotomy with VATS. The VATS through two small skin incisions in the axillary region provides a good view without requiring elevation of the scapula with extensile muscle dissection and rib resection. There was no complication without partial lobectomy due to pleural adhesion during the perioperative period. Currently, at 1 year after operation, the patient has no back pain with neurologically normal findings and no inflammation findings (CRP was 0.01 mg/dl). Although the operating field of the upper thoracic level in the lateral approach is generally deep and narrow, the VATS provides a good view and allows us to perform adequate debridement and bone fusion at the T2/3 level with a less invasive approach than those previously described anterior or laterally or posterior approach

First clinical experience with posterior lumbar interbody fusion using a thermal-sprayed silver-containing hydroxyapatite-coated cage

Abstract Background To investigate the possibility of silver (Ag)-induced adverse events and the degree of bone fusion in posterior lumbar interbody fusion surgery using an Ag-containing hydroxyapatite (HA) lumbar interbody cage. Methods An Ag-HA cage consisting of highly osteoconductive HA interfused with Ag was developed, and we applied it clinically at three university-affiliated hospitals from April 2020 to December 2020. During the 12-month postoperative observation period, Ag-related adverse events, neuropathy, and postoperative complications were investigated as indicators of safety, while clinical improvement and the fusion status were investigated as indicators of efficacy. Clinical improvement was defined as improvement beyond the minimum clinically important difference (MCID) in the numerical rating scale (NRS; 1.6) for low back and lower limb pain and the Oswestry Disability Index (ODI; 12.8). Results We performed lumbar interbody fusion using an Ag-HA cage for 48 patients (female, n = 25; mean age, 67.5 years). The mean preoperative NRS was 6.4 (standard deviation, 1.9), while the mean preoperative ODI was 44 [12]. No adverse effects (i.e., argyria) were identified during the 12-month observation period. Surgical site infection occurred in one case, although the implant was preserved via immediate debridement. In total, 39 (81%) participants showed clinical improvement beyond MCID for both NRS and ODI. Bone fusion was achieved at 45 levels (88%) at 6 months and 48 levels (91%) at 12 months postoperatively. Conclusions The results of this study suggest that Ag-HA cages can be safely used in spinal fusion procedures and have the potential to prevent postoperative infections, prevent deterioration of the quality of life, and result in favorable outcomes. Larger-scale and longer-term follow-up studies will be required to corroborate these conclusions. Trial registration UMIN 000039964 (date: April 01, 2020)