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1.
Parasitol Res ; 118(9): 2509-2521, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31377908

ABSTRACT

In this study, we characterize the HIP-55 protein in the mosquito Aedes aegypti for the first time. HIP-55 is a 55-kDa HPK1-interacting protein that is also called SH3P7. HIP-55 constitutively binds HPK1 'via' an HPK1 proline-rich motif 2(PR2) through its C-terminal SH3 domain. HIP-55 critically interacts with ZAP-70, and this interaction was induced by TCR signalling. ZAP-70 phosphorylated HIP-55 at Tyr-334 and Tyr-344 in vitro and in vivo. In our previous findings, AaZAP gene expression strongly proved that AaZAP-70 was involved in immunity-like functions in mosquito. Northern blot analysis of HIP-55 mRNA expression confirmed that it is only expressed in the abdomen and haemocyte tissues; this prediction correlates 100% and a polyclonal antibody also confirmed its localization in haemocytes and the abdomen. We prepared extracts to show the cytoplasmic expression (CE) of this protein. Previous results had proven that this protein is secreted from the cytoplasm; thus, we confirmed here that the protein is a cytoplasmic adaptor protein in mosquitoes and mammalian systems. Furthermore, our polyclonal antibody against HIP-55 also demonstrated that this protein is found in haemocytes and abdomen tissues, which assumes that the protein may be involved in phagocytic-like functions. RNAi (siRNA) silencing studies were used to degrade mosquito HIP-55; however, silencing only slightly affected the HIP-55 sequence and the gene transcriptional level. To characterize this protein, we cloned 609 bp from the 1.6-kb full-length cDNA using a pET28 vector for polyclonal antibody production. Graphical abstract.


Subject(s)
Aedes/immunology , Microfilament Proteins/metabolism , Protein Serine-Threonine Kinases/metabolism , src Homology Domains/physiology , Animals , Hemocytes/metabolism , Microfilament Proteins/genetics , Phosphorylation , Protein Binding , RNA Interference , RNA, Messenger/genetics , RNA, Small Interfering/genetics , Signal Transduction , ZAP-70 Protein-Tyrosine Kinase/metabolism , src Homology Domains/genetics
2.
Indian J Gastroenterol ; 33(2): 157-60, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24105667

ABSTRACT

UNLABELLED: High resolution esophageal manometry (HREM) has been interpreted all along by visual interpretation of color plots until the recent introduction of Chicago classification which categorises HREM using objective measurements. It compares HREM diagnosis of esophageal motor disorders by visual interpretation and Chicago classification. Using software Trace 1.2v, 77 consecutive tracings diagnosed by visual interpretation were re-analyzed by Chicago classification and findings compared for concordance between the two systems of interpretation. STATISTICAL ANALYSIS: Kappa agreement rate between the two observations was determined. There were 57 males (74 %) and cohort median age was 41 years (range: 14-83 years). Majority of the referrals were for gastroesophageal reflux disease, dysphagia and achalasia. By "intuitive" visual interpretation, the tracing were reported as normal in 45 (58.4 %), achalasia 14 (18.2 %), ineffective esophageal motility 3 (3.9 %), nutcracker esophagus 11 (14.3 %) and nonspecific motility changes 4 (5.2 %). By Chicago classification, there was 100 % agreement (Kappa 1) for achalasia (type 1: 9; type 2: 5) and ineffective esophageal motility ("failed peristalsis" on visual interpretation). Normal esophageal motility, nutcracker esophagus and nonspecific motility disorder on visual interpretation were reclassified as rapid contraction and esophagogastric junction (EGJ) outflow obstruction by Chicago classification. Chicago classification identified distinct clinical phenotypes including EGJ outflow obstruction not identified by visual interpretation. A significant number of unclassified HREM by visual interpretation were also classified by it.


Subject(s)
Esophageal Motility Disorders/classification , Esophageal Motility Disorders/diagnosis , Image Enhancement/methods , Image Interpretation, Computer-Assisted/methods , Manometry/methods , Adolescent , Adult , Aged , Female , Gastroesophageal Reflux/diagnosis , Humans , Male , Middle Aged , Software , Young Adult
3.
Hum Reprod ; 24(9): 2124-32, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19493874

ABSTRACT

BACKGROUND: Sono-automatic volume calculation (SonoAVC) automatically identifies and measures the dimensions of hypoechogenic areas within datasets acquired using three-dimensional ultrasound. The objective of this study was to evaluate the predictive value of automated antral follicle counts according to their relative sizes in women undergoing assisted reproduction treatment (ART). METHODS: A total of 156 subjects aged < or =40 years with a baseline FSH < or =15 IU that undergo their first cycle of ART were prospectively recruited. SonoAVC was used to measure the datasets and record the number of antral follicles measuring < or =9 mm in diameter. These follicles were then grouped into subsets according to their relative sizes: < or =2.0, 2.1-4.0, 4.1-6.0, 6.1-8.0 and 8.1-9.0 mm. The primary outcome was viable pregnancy confirmed on ultrasound 5 weeks following embryo transfer. RESULTS: A total of 142 subjects were included for analysis of primary end-point. Those subjects who conceived had significantly more antral follicles measuring < or =2 (P = 0.041) and 2.1-4.0 mm (P < 0.001) than those who had unsuccessful treatment. There were no significant differences between the groups in the number of antral follicles measuring 4.1-6.0 (P = 0.191), 6.1-8.0 (P = 0.203) and 8.1-9.0 mm (P = 0.601). Multiple logistic regression showed that antral follicles measuring 2.1-4.0 mm were an independent predictor of pregnancy [Exp(B) = 1.234, 95% CI = 1.092-1.491; P = 0.004; AUC = 0.693]. CONCLUSION: SonoAVC provides automated measures of antral follicle number and size. Using this technique, the number of antral follicles measuring 2.1-4.0 mm in diameter is an independent, significant predictor of pregnancy following in vitro fertilization treatment.


Subject(s)
Follicular Phase , Ovarian Follicle/diagnostic imaging , Reproductive Techniques, Assisted , Adult , Embryo Transfer , Female , Humans , Ovarian Follicle/ultrastructure , Predictive Value of Tests , Regression Analysis , Treatment Outcome , Ultrasonography
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