Hyaluronan- and RNA-binding deubiquitinating enzymes of USP17 family members associated with cell viability

BACKGROUND: Protein degradation by the ubiquitin system plays a crucial role in numerous cellular signaling pathways. Deubiquitination, a reversal of ubiquitination, has been recognized as an important regulatory step in the ubiquitin-dependent degradation pathway. RESULTS: While identifying putative ubiquitin specific protease (USP) enzymes that contain a conserved Asp (I) domain in humans, 4 USP17 subfamily members, highly homologous to DUB-3, have been found (USP17K, USP17L, USP17M, and USP17N), from human chorionic villi. Expression analysis showed that USP17 transcripts are highly expressed in the heart, liver, and pancreas and are expressed moderately in various human cancerous cell lines. Amino acid sequence analysis revealed that they contain the highly conserved Cys, His, and Asp domains which are responsible for the deubiquitinating activity. Biochemical enzyme assays indicated that they have deubiquitinating activity. Interestingly, the sequence analysis showed that these proteins, with exception of USP17N, contain the putative hyaluronan/RNA binding motifs, and cetylpyridinium chloride (CPC)-precipitation analysis confirmed the association between these proteins and intracellular hyaluronan and RNA. CONCLUSION: Here, we report that the overexpression of these proteins, with exception of USP17N, leads to apoptosis, suggesting that the hyaluronan and RNA binding motifs in these enzymes play an important role in regulating signal transduction involved in cell death

Actin Cytoskeleton and Golgi Involvement in Barley stripe mosaic virus Movement and Cell Wall Localization of Triple Gene Block Proteins.

Barley stripe mosaic virus (BSMV) induces massive actin filament thickening at the infection front of infected Nicotiana benthamiana leaves. To determine the mechanisms leading to actin remodeling, fluorescent protein fusions of the BSMV triple gene block (TGB) proteins were coexpressed in cells with the actin marker DsRed: Talin. TGB ectopic expression experiments revealed that TGB3 is a major elicitor of filament thickening, that TGB2 resulted in formation of intermediate DsRed:Talin filaments, and that TGB1 alone had no obvious effects on actin filament structure. Latrunculin B (LatB) treatments retarded BSMV cell-to-cell movement, disrupted actin filament organization, and dramatically decreased the proportion of paired TGB3 foci appearing at the cell wall (CW). BSMV infection of transgenic plants tagged with GFP-KDEL exhibited membrane proliferation and vesicle formation that were especially evident around the nucleus. Similar membrane proliferation occurred in plants expressing TGB2 and/or TGB3, and DsRed: Talin fluorescence in these plants colocalized with the ER vesicles. TGB3 also associated with the Golgi apparatus and overlapped with cortical vesicles appearing at the cell periphery. Brefeldin A treatments disrupted Golgi and also altered vesicles at the CW, but failed to interfere with TGB CW localization. Our results indicate that actin cytoskeleton interactions are important in BSMV cell-to-cell movement and for CW localization of TGB3

A genome-wide Asian genetic map and ethnic comparison: The GENDISCAN study

<p>Abstract</p> <p>Background</p> <p>Genetic maps provide specific positions of genetic markers, which are required for performing genetic studies. Linkage analyses of Asian families have been performed with Caucasian genetic maps, since appropriate genetic maps of Asians were not available. Different ethnic groups may have different recombination rates as a result of genomic variations, which would generate misspecification of the genetic map and reduce the power of linkage analyses.</p> <p>Results</p> <p>We constructed the genetic map of a Mongolian population in Asia with CRIMAP software. This new map, called the GENDISCAN map, is based on genotype data collected from 1026 individuals of 73 large Mongolian families, and includes 1790 total and 1500 observable meioses. The GENDISCAN map provides sex-averaged and sex-specific genetic positions of 1039 microsatellite markers in Kosambi centimorgans (cM) with physical positions. We also determined 95% confidence intervals of genetic distances of the adjacent marker intervals.</p> <p>Genetic lengths of the whole genome, chromosomes and adjacent marker intervals are compared with those of Rutgers Map v.2, which was constructed based on Caucasian populations (Centre d'Etudes du Polymorphisme Humain (CEPH) and Icelandic families) by mapping methods identical to those of the GENDISCAN map, CRIMAP software and the Kosambi map function. Mongolians showed approximately 1.9 fewer recombinations per meiosis than Caucasians. As a result, genetic lengths of the whole genome and chromosomes of the GENDISCAN map are shorter than those of Rutgers Map v.2. Thirty-eight marker intervals differed significantly between the Mongolian and Caucasian genetic maps.</p> <p>Conclusion</p> <p>The new GENDISCAN map is applicable to the genetic study of Asian populations. Differences in the genetic distances between the GENDISCAN and Caucasian maps could facilitate elucidation of genomic variations between different ethnic groups.</p

Double-layered TiO2 photoelectrode with particulate structure prepared by one-step soaking method

Nanostructured TiO2 films with double-layered structure are prepared by a facile one-step soaking method. We have investigated the morphology of nanostructured TiO2 films by the reaction time of the soaking method, which has an effect on the thickness and layered structure of the TiO2 films. The TiO2 films prepared by this method have a unique double-layered structure, which is composed of a dense TiO2 bottom layer and TiO2 particulates on the bottom layer. By manipulating the reaction time of the soaking method, control of TiO2 particulate formation on the surface of the dense TiO2 bottom layer is possible. The double-layered structure of nanostructured TiO2 films is effective for achieving sufficient adsorption of Sb2S3 sensitizer and light scattering effect of photoelectrodes for inorganic sensitized solar cells, which induces the enhancement of short circuit current of solar cell devices. Our solar cell device, using a doublelayered TiO2 film with particulate structure as a photoelectrode, exhibited JSC, VOC, FF, and η values of 12.94 mA/cm2, 498 mV, 57.0%, and 3.67%, respectively. © 2014 Optical Society of America.1

Enhanced cardiac expression of two isoforms of matrix metalloproteinase-2 in experimental diabetes mellitus.

BackgroundDiabetic cardiomyopathy (DM CMP) is defined as cardiomyocyte damage and ventricular dysfunction directly associated with diabetes independent of concomitant coronary artery disease or hypertension. Matrix metalloproteinases (MMPs), especially MMP-2, have been reported to underlie the pathogenesis of DM CMP by increasing extracellular collagen content.PurposeWe hypothesized that two discrete MMP-2 isoforms (full length MMP-2, FL-MMP-2; N-terminal truncated MMP-2, NTT-MMP-2) are induced by high glucose stimulation in vitro and in an experimental diabetic heart model.MethodsRat cardiomyoblasts (H9C2 cells) were examined to determine whether high glucose can induce the expression of the two isoforms of MMP-2. For the in vivo study, we used the streptozotocin-induced DM mouse heart model and age-matched controls. The changes of each MMP-2 isoform expression in the diabetic mice hearts were determined using quantitative real-time polymerase chain reaction (qRT-PCR). Immunohistochemical stains were conducted to identify the location and patterns of MMP-2 isoform expression. Echocardiography was performed to compare and analyze the changes in cardiac function induced by diabetes.ResultsQuantitative RT-PCR and immunofluorescence staining showed that the two MMP-2 isoforms were strongly induced by high glucose stimulation in H9C2 cells. Although no definite histologic features of diabetic cardiomyopathy were observed in diabetic mice hearts, left ventricular systolic dysfunction was determined by echocardiography. Quantitative RT-PCR and IHC staining showed this abnormal cardiac function was accompanied with the increases in the mRNA levels of the two isoforms of MMP-2 and related to intracellular localization.ConclusionTwo isoforms of MMP-2 were induced by high glucose stimulation in vitro and in a Type 1 DM mouse heart model. Further study is required to examine the role of these isoforms in DM CMP

Effect of pulmonary rehabilitation on patients with acute COVID-19: a single-center case series

The coronavirus disease 2019 (COVID-19) pandemic has been ongoing for more than 2 years. Many patients who recover from severe acute respiratory syndrome coronavirus 2 infection continue to have aftereffects such as dyspnea and fatigue, which may lead to functional decline. Therefore, the need for managing these symptoms using methods such as pulmonary rehabilitation (PR) has emerged. The purpose of this study was to report the effectiveness of PR in five patients with acute COVID-19. PR was performed in patients with persistent dyspnea and oxygen demand after COVID-19. All five patients were able to maintain an independent functional status before COVID-19. However, after acute COVID-19, they were unable to walk independently and needed assistance for activities of daily living due to dyspnea and fatigue. Therefore, they were referred to rehabilitation units, and PR was performed. The modified Medical Research Council dyspnea scale, maximal expiratory pressure (MEP), 6-minute walking test, forced vital capacity, and grip strength were assessed before and after PR, and the results were compared. After PR, the parameters improved, except for the MEP in one patient (patient 3) and the grip strength in another patient (patient 4). After PR, two out of five patients returned to work and the other three returned home. Therefore, we conclude that PR is necessary for patients with acute COVID-19 with activity limitations

N-Nitrosamine inhibition by strawberry, garlic, kale, and the effects of nitrite-scavenging and Nnitrosamine formation by functional compounds in strawberry and garlic, Food Control

Abstract High nitrate and amine-rich food intake may result in an increased risk of endogenous formation of carcinogenic N-nitroso compounds (NOCs). We studied the eVects of whole strawberries, garlic and kale on the formation of N-nitrosodimethylamine (NDMA) in a traditional Korean diet (amine and nitrate-rich) in simulated saliva and gastric conditions. The addition of whole strawberries, kale and garlic juices in the diet reduced NDMA formation by amine and nitrate. Strawberries and garlic juice were more eVective than kale. To identify active compound, isolated fractions were prepared from strawberries or garlic juice samples by preparative liquid chromatography. Inhibition of NDMA formation and nitrite-scavenging ability of each fraction (strawberries: S1-S4; garlic juice: A1-A5) was tested. Fraction A4 inhibited NDMA formation by 50.5 § 5.4% and increased the eVect of nitrite-scavenging by 72.3 § 8.7%. Fractions A3 and A4 were identiWed as 1,2-benzendicarboxy acid (A3) and S-oxodially disulWde (A4) compounds by mass spectrophotometry and 13 C nuclear magnetic resonance