scholarly journals Characterization of THSD7A-antibodies not binding to glomerular THSD7A in a patient with diabetes mellitus but no membranous nephropathy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Linda Reinhard ◽  
Cindy Thomas ◽  
Maya Machalitza ◽  
Erik Lattwein ◽  
Lothar S. Weiss ◽  
...  
Keyword(s):  
Western Blot ◽  
Membranous Nephropathy ◽  
Kidney Biopsy ◽  
Reducing Conditions ◽  
Igg3 Subclass ◽  
Antibody Levels ◽  
First Time

AbstractMembranous nephropathy (MN) is an autoimmune disease caused by autoantibodies against the podocyte antigens phospholipase A2 receptor 1 (PLA2R1) and thrombospondin type 1 domain containing protein 7A (THSD7A) in 80% and 2–3% of patients, respectively. THSD7A antibodies are considered to be pathogenic and highly specific for MN patients. Using an indirect immunofluorescence test (IIFT) we detected THSD7A-antibodies (titre 1:10) in the serum of a patient with high proteinuria who, however, in the kidney biopsy was diagnosed with diabetic nephropathy and MN was excluded as a possible cause of proteinuria. Different immunofluorescence assays and Western blot techniques using recombinant THSD7A (rTHSD7A) or THSD7A from different human tissues revealed that the circulating THSD7A-autoantibodies were only of the IgG3 subclass. The patient serum reacted exclusively with rTHSD7A and only when the antigen was present in reducing Western blot conditions, or on formaldehyde-fixed cells for the IIFT. Our findings show for the first time the existence of circulating THSD7A-antibodies recognizing denatured/reduced rTHSD7A, which do not react with glomerular THSD7A in vivo and are thus presumptively non-pathogenic. As a consequence, kidney biopsy or Western blot analyses of THSD7A under non-reducing conditions should be performed to confirm the diagnosis of THSD7A-associated MN, especially in cases with low THSD7A-antibody levels in the IIFT.

scholarly journals Synthesis and Characterization of Exopolysaccharide Encapsulated PCL/Gelatin Skin Substitute for Full-Thickness Wound Regeneration

Polymers ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 854
Author(s):  
Ahmad Hivechi ◽  
Peiman Brouki Milan ◽  
Khashayar Modabberi ◽  
Moein Amoupour ◽  
Kaveh Ebrahimzadeh ◽  
...  
Keyword(s):  
Cell Activity ◽  
Average Diameter ◽  
Skin Substitute ◽  
Full Thickness ◽  
Skin Integrity ◽  
Hematoxylin And Eosin ◽  
First Time ◽  
Full Thickness Wound

Loss of skin integrity can lead to serious problems and even death. In this study, for the first time, the effect of exopolysaccharide (EPS) produced by cold-adapted yeast R. mucilaginosa sp. GUMS16 on a full-thickness wound in rats was evaluated. The GUMS16 strain’s EPS was precipitated by adding cold ethanol and then lyophilized. Afterward, the EPS with polycaprolactone (PCL) and gelatin was fabricated into nanofibers with two single-needle and double-needle procedures. The rats’ full-thickness wounds were treated with nanofibers and Hematoxylin and eosin (H&E) and Masson’s Trichrome staining was done for studying the wound healing in rats. Obtained results from SEM, DLS, FTIR, and TGA showed that EPS has a carbohydrate chemical structure with an average diameter of 40 nm. Cell viability assessments showed that the 2% EPS loaded sample exhibits the highest cell activity. Moreover, in vivo implantation of nanofiber webs on the full-thickness wound on rat models displayed a faster healing rate when EPS was loaded into a nanofiber. These results suggest that the produced EPS can be used for skin tissue engineering applications.

P.3.d.011 In vitro and in vivo characterization of glycine transporter type-1 by two inhibitors, Org-24461 and NFPS

2006 ◽  
Vol 16 ◽  
pp. S435 ◽  
Author(s):  
B. Marko ◽  
G. Szabo ◽  
S. Udvari ◽  
M. Agoston ◽  
E. Szabo ◽  
...  
Keyword(s):  
Glycine Transporter ◽  
In Vivo Characterization

scholarly journals Identification and characterization of molecular interactions between mortalin/mtHsp70 and HSP60

2005 ◽  
Vol 391 (2) ◽  
pp. 185-190 ◽  
Author(s):  
Renu Wadhwa ◽  
Syuichi Takano ◽  
Kamaljit Kaur ◽  
Satoshi Aida ◽  
Tomoko Yaguchi ◽  
...  
Keyword(s):  
Heat Shock ◽  
Molecular Interactions ◽  
Heat Shock Protein 60 ◽  
Hsp60 Expression ◽  
Mitochondrial Hsp70 ◽  
Shock Proteins ◽  
First Time

Mortalin/mtHsp70 (mitochondrial Hsp70) and HSP60 (heat-shock protein 60) are heat-shock proteins that reside in multiple subcellular compartments, with mitochondria being the predominant one. In the present study, we demonstrate that the two proteins interact both in vivo and in vitro, and that the N-terminal region of mortalin is involved in these interactions. Suppression of HSP60 expression by shRNA (short hairpin RNA) plasmids caused the growth arrest of cancer cells similar to that obtained by suppression of mortalin expression by ribozymes. An overexpression of mortalin, but not of HSP60, extended the in vitro lifespan of normal fibroblasts (TIG-1). Taken together, this study for the first time delineates: (i) molecular interactions of HSP60 with mortalin; (ii) their co- and exclusive localizations in vivo; (iii) their involvement in tumorigenesis; and (iv) their functional distinction in pathways involved in senescence.

In Vivo Characterization of Corneal Changes in a Type 1 Diabetic Animal Model

2019 ◽  
Vol 45 (3) ◽  
pp. 823-832
Author(s):  
Miguel Caixinha ◽  
Pedro Oliveira ◽  
Inês D. Aires ◽  
António Francisco Ambrósio ◽  
Ana Raquel Santiago ◽  
...  
Keyword(s):  
Animal Model ◽  
Diabetic Animal ◽  
In Vivo Characterization

scholarly journals Characterization of the complete genomes of Camelus dromedarius papillomavirus types 1 and 2

2011 ◽  
Vol 92 (8) ◽  
pp. 1769-1777 ◽  
Author(s):  
A. E. Ure ◽  
A. K. Elfadl ◽  
A. I. Khalafalla ◽  
A. A. R. Gameel ◽  
J. Dillner ◽  
...  
Keyword(s):  
Phylogenetic Analyses ◽  
Diagnostic Methods ◽  
Camelus Dromedarius ◽  
Bovine Papillomavirus ◽  
Lower Jaw ◽  
The Family ◽  
Complete Genomes ◽  
First Time

Camel papillomatosis has been described previously, but the genome of the suspected papillomavirus (PV) has not been identified. An outbreak of papillomatosis occurred in a dromedary farm of 55 animals in Sudan during August 2009. The disease was only present in young animals aged about 3–7 months, of which 44 % (11/25) were affected with lesions, mainly on the lips and lower jaw. This study reports for the first time the complete genomes of Camelus dromedarius papillomavirus types 1 (CdPV1) and 2 (CdPV2), isolated from a cauliflower-like nodule and a round oval raised nodule, respectively. Pairwise comparisons of their L1 nucleotide sequences revealed 69.2 % identity, and phylogenetic analyses suggested that these two PV types are grouped within the genus Deltapapillomavirus. Both viruses were isolated from fibropapillomas, although no putative E5 proteins homologous to that of bovine papillomavirus type 1 were identified. The genetic information will be useful for evolutionary studies of the family Papillomaviridae, as well as for the development of diagnostic methods for surveillance of the disease in dromedaries.

scholarly journals Characterization of the Novel Mitochondrial Genome Segregation Factor TAP110 in Trypanosoma brucei

2020 ◽  
Author(s):  
Simona Amodeo ◽  
Ana Kalichava ◽  
Albert Fradera-Sola ◽  
Eloïse Bertiaux-Lequoy ◽  
Paul Guichard ◽  
...  
Keyword(s):  
Mitochondrial Genome ◽  
Trypanosoma Brucei ◽  
Protozoan Parasite ◽  
Eukaryotic Cell ◽  
The Novel ◽  
Detergent Treatment ◽  
Summary Statement ◽  
First Time

AbstractProper mitochondrial genome inheritance is key for eukaryotic cell survival, however little is known about the molecular mechanism controlling this process. Trypanosoma brucei, a protozoan parasite, contains a singular mitochondrial genome aka kinetoplast DNA (kDNA). kDNA segregation requires anchoring of the genome to the basal body via the tripartite attachment complex (TAC). Several components of the TAC as well as their assembly have been described, it however remains elusive how the TAC connects to the kDNA. Here, we characterize the TAC associated protein TAP110 and for the first time use ultrastructure expansion microscopy in trypanosomes to reveal that TAP110 is the currently most proximal kDNA segregation factor. The kDNA proximal positioning is also supported by RNAi depletion of TAC102, which leads to loss of TAP110 at the TAC. Overexpression of TAP110 leads to expression level changes of several mitochondrial proteins and a delay in the separation of the replicated kDNA networks. In contrast to other kDNA segregation factors TAP110 remains only partially attached to the flagellum after DNAse and detergent treatment and can only be solubilized in dyskinetoplastic cells, suggesting that interaction with the kDNA might be important for stability of the TAC association. Furthermore, we demonstrate that the TAC, but not the kDNA, is required for correct TAP110 localization in vivo and suggest that TAP110 might interact with other proteins to form a >669 kDa complex.Summary StatementTAP110 is a novel mitochondrial genome segregation factor in Trypanosoma brucei that associates with the previously described TAC component TAC102. Ultrastructure expansion microscopy reveals its proximal position to the kDNA.

scholarly journals Analysis of tissue PLA2R efficacy in evaluating the prognosis of idiopathic membranous nephropathy in the background of different serum anti-PLA2R levels

2020 ◽  
Author(s):  
Yuemeng Sun ◽  
Ping Lan ◽  
Jie Feng ◽  
Zhigang Wang ◽  
Chao Liu ◽  
...  
Keyword(s):  
Antibody Titer ◽  
Membranous Nephropathy ◽  
Kidney Biopsy ◽  
Clinical Presentation ◽  
Remission Rate ◽  
Antigen Expression ◽  
Predominant Role ◽  
Positive Correlation ◽  
Disease Prognosis ◽  
Antibody Levels

Objective To verify serum PLA2R antibody and glomerular PLA2R antigen expression in membranous nephropathy as well as to explore their relationship with clinical presentation and disease prognosis. Methods We retrospectively analyzed 155 patients clinical figures who were diagnosed with primary membranous nephropathy by kidney biopsy. Patients were divided into 6 groups according to their serum PLA2R antibody or glomerular PLA2R antigen positiveness and the level of serum PLA2R antibody titer. Both clinical features and pathological characteristics were recorded, and remission rate as well as time to response were compared among groups. Correlation between clinical figures and titer of PLA2R antibody or semi-quantity of PLA2R antigen were detected. Results Among patients with positive serum PLA2R antibody and tissue PLA2R antigen, higher baseline PLA2R antibody levels were associated with lower remission rate and longer remission time. A positive correlation between time to partial remission and serum PLA2R antibody titer was found. Among patients with serum PLA2R antibody titer <150U/L, there were shorter remission time in negative tissue PLA2R antigen group compared with positive tissue PLA2R antigen, and a positive correlation between time to complete remission and semi-quantity of tissue PLA2R antigen was found. Conclusion Both glomerular PLA2R antigen and serum PLA2R antibody play a role in disease presentation and prognosis in primary membranous nephropathy. Glomerular PLA2R antigen has a major role on disease prognosis when serum PLA2R antibody titer is less than 150U/L, while serum PLA2R antibody has predominant role in MN prognosis when serum PLA2R antibody titer is above 150 U/L.

scholarly journals Partial Sperm beta1 Integrin Subunit Deletion Proves Its Involvement in Mouse Gamete Adhesion/Fusion

2020 ◽  
Vol 21 (22) ◽  
pp. 8494
Author(s):  
Virginie Barraud-Lange ◽  
Côme Ialy-Radio ◽  
Céline Chalas ◽  
Isabelle Holtzmann ◽  
Jean-Philippe Wolf ◽  
...  
Keyword(s):  
Western Blot ◽  
Germ Cells ◽  
Blot Analysis ◽  
Integrin Subunit ◽  
Perivitelline Space ◽  
Male Germ Cells ◽  
Beta1 Integrin ◽  
First Time

We have previously shown, using antibodies, that the sperm alpha6beta1 integrin is involved in mouse gamete fusion in vitro. Here we report the conditional knockdown of the sperm Itgb1 gene. It induced a drastic failure of sperm fusogenic ability with sperm accumulation in the perivitelline space of in vitro inseminated oocytes deleted or not for the Itgb1 gene. These data demonstrate that sperm, but not oocyte, beta1 integrin subunit is involved in gamete adhesion/fusion. Curiously, knockdown males were fertile in vivo probably because of the incomplete Cre-mediated deletion of the sperm Itgb1 floxed gene. Indeed, this was shown by Western blot analysis and confirmed by both the viability and litter size of pups obtained by mating partially sperm Itgb1 deleted males with females producing completely deleted Itgb1 oocytes. Because of the total peri-implantation lethality of Itgb1 deletion in mice, we assume that sperm that escaped the Itgb1 excision seemed to be preferentially used to fertilize in vivo. Here, we showed for the first time that the deletion, even partial, of the sperm Itgb1 gene makes the sperm unable to normally fertilize oocytes. However, to elucidate the question of the essentiality of its role during fertilization, further investigations using a mouse expressing a recombinase more effective in male germ cells are necessary.

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