scholarly journals A Rapid and Simple UHPLC-MS/MS Method for Quantification of Plasma Globotriaosylsphingosine (lyso-Gb3)

Molecules ◽  
2021 ◽  
Vol 26 (23) ◽  
pp. 7358
Author(s):  
Alessandro Perrone ◽  
Susan Mohamed ◽  
Vincenzo Donadio ◽  
Rocco Liguori ◽  
Manuela Contin
Keyword(s):  
Plasma Concentrations ◽  
Control Group ◽  
P Value ◽  
Healthy Controls ◽  
Lysosomal Storage ◽  
Limit Of Quantification ◽  
Accuracy And Precision ◽  
Ionization Method ◽  
Pre Treatment ◽  
Gla Gene

Fabry disease (FD) is a rare X-linked lysosomal storage disorder caused by α-galactosidase A gene (GLA) mutations, resulting in loss of activity of the lysosomal hydrolase, α-galactosidase A (α-Gal A). As a result, the main glycosphingolipid substrates, globotriaosylceramide (Gb3) and globotriaosylsphingosine (lyso-Gb3), accumulate in plasma, urine, and tissues. Here, we propose a simple, fast, and sensitive method for plasma quantification of lyso-Gb3, the most promising secondary screening target for FD. Assisted protein precipitation with methanol using Phree cartridges was performed as sample pre-treatment and plasma concentrations were measured using UHPLC-MS/MS operating in MRM positive electrospray ionization. Method validation provided excellent results for the whole calibration range (0.25–100 ng/mL). Intra-assay and inter-assay accuracy and precision (CV%) were calculated as <10%. The method was successfully applied to 55 plasma samples obtained from 34 patients with FD, 5 individuals carrying non-relevant polymorphisms of the GLA gene, and 16 healthy controls. Plasma lyso-Gb3 concentrations were larger in both male and female FD groups compared to healthy subjects (p < 0.001). Normal levels of plasma lyso-Gb3 were observed for patients carrying non-relevant mutations of the GLA gene compared to the control group (p = 0.141). Dropping the lower limit of quantification (LLOQ) to 0.25 ng/mL allowed us to set the optimal plasma lyso-Gb3 cut-off value between FD patients and healthy controls at 0.6 ng/mL, with a sensitivity of 97.1%, specificity of 100%, and accuracy of 0.998 expressed by the area under the ROC curve (C.I. 0.992 to 1.000, p-value < 0.001). Based on the results obtained, this method can be a reliable tool for early phenotypic assignment, assessing diagnoses in patients with borderline GalA activity, and confirming non-relevant mutations of the GLA gene.

scholarly journals Outcome of vaginal sildinafil in assisted reproductive technology cycles

2021 ◽  
Vol 10 (2) ◽  
pp. 575
Author(s):  
Avnika Kapoor ◽  
Garima Sharma ◽  
Rita Bakshi
Keyword(s):  
Uterine Artery ◽  
Endometrial Thickness ◽  
Study Groups ◽  
Control Group ◽  
P Value ◽  
Surrogate Mothers ◽  
Endometrial Pattern ◽  
Study Population ◽  
Pre Treatment ◽  
And Control

Background: The present study assessed the role of sildenafil in endometrial blood flow and successful pregnancy in IVF done in surrogate mothers.Methods: In the present study surrogate mothers were included. Thirty patients were randomized to receive sildenafil 25 mg thrice a day vaginally in addition to standard drugs and technique and another 30 were not given sildenafil.Results: Mean age, anthropometry, duration of infertility and pre-treatment endometrial thickness was similar in the two study groups.  After treatment completion, it was observed that the endometrial pattern in ultrasound was similar in the two study groups (p value=0.58). Heterogenic endometrial pattern was observed in 6.7% of the Sildenafil patients and 3.3% in the control patients, while echogenic pattern was seen in 10% of the sildenafil patients and 6.7% of the control patients. Similarly, endometrial thickness was 10.2±1.7 and 9.7±1.8 mm in sildenafil and control group respectively, p value=0.62. Using doppler ultrasound, uterine artery PI was significantly lower in Sildenafil group patients as compared to control group patients. Similarly, we found uterine artery RI was also significantly lower in the Sildenafil group patients as compared to control group patients. We followed the patients and found that clinical pregnancy rate was significantly higher among Sildenafil group (60%) as compared to control group (26.6%), p value<0.05.Conclusions: Vaginal sildenafil resulted in significantly higher pregnancy rates in our study population. The uterine artery PI and RI were significantly lower in patients taking sildenafil.

Soluble programmed death ligand-1(sPD-L1) is elevated in aggressive prostate cancer disease among African men

2021 ◽  
Author(s):  
PAUL KATONGOLE ◽  
Obondo J. Sande ◽  
Steven J Reynolds ◽  
Moses Joloba ◽  
Henry Kajumbula ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Plasma Concentrations ◽  
Enzyme Linked Immunosorbent Assay ◽  
P Value ◽  
Healthy Controls ◽  
Cancer Disease ◽  
Programmed Death Ligand 1 ◽  
Programmed Death ◽  
Median Plasma ◽  
Programmed Death 1

Abstract Background The programmed death 1 (PD1)/programmed death-ligand 1 (PDL1) targeted immunotherapies have become a new mode of treatment for several tumours; however, there is limited evidence on the expression and prognostic value of PD1/PDL1 in prostate cancer, especially in African men. Methods The plasma concentrations of PD-L1/PD1 were assessed using Enzyme-Linked Immunosorbent Assay in patients with prostate cancer and normal healthy controls at the Uganda Cancer Institute. The association between plasma PD-L1/PD1 concentration levels and PSA levels, Gleason scores, age, and Body mass index were determined. Results We found significant differences in the median plasma concentrations of PD-L1 and PD-1 immune checkpoint molecules between Prostate cancer cases and normal healthy controls of (0.285 vs 0.035) p-value 0.001 and (0.596 vs 0.355) p-value 0.017, respectively. We found no significant association between age, plasma PSA levels, BMI and Gleason scores, and PD-1 among patients with prostate cancer and controls. However, elevated levels of PD-L1 were significantly associated with raised Gleason scores among patients with prostate cancer with a p-value of <0.001. Conclusions Elevated PD-L1 levels were statistically significantly linked to high Gleason scores. These results may guide clinicians in assessing the prognosis of patients individually and selecting suitable patients that will make favorable candidates for anti-PD-L1 immunotherapy.

MO008B LYMPHOCYTE SUBSET CHANGES IN PRIMARY MEMBRANEOUS NEPHROPATHY

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Nooshin Dalili ◽  
Fatemeh Pour-Reza Gholi ◽  
Katayoun Hasanzadeh ◽  
Sara Assadiasl
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Standard Treatment ◽  
Control Group ◽  
P Value ◽  
Healthy Controls ◽  
Memory B Cell ◽  
Wide Range ◽  
Significant Difference

Abstract Background and Aims Primary membranous nephropathy (PMN) is the most common cause of nephrotic syndrome (NS) in adults. Rituximab a chimeric monoclonal anti-CD20 antibody has been supposed to eliminate autoantibody-producing B cells via direct signaling, complement-mediated cytotoxicity (CMC), and antibody-dependent cellular cytotoxicity (ADCC). According to the fact that a wide range of B lymphocytes may carry this marker, we aimed to identify which subset is more (or less) frequent in PMN patients and which one is more affected by rituximab administration. Method Three groups were enrolled in the present study. They included a healthy control group and two patient groups having the clinical, laboratory, and pathological diagnostic criteria of PMN, comprising either patients on standard treatment or patients on standard treatment plus rituximab. The latter group was studied just before receiving rituximab (pre-rituximab) and two months later (post rituximab). Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-hypaque (inno-train, Germany) gradient. Afterward, cells were adjusted to a concentration of 1 _ 106 cells/mL and stained with mouse antihuman CD19-Cy5-conjugated antibody (Cytognos, Spain), mouse antihuman CD24-PE-conjugated antibody (Cytognos, Spain), and mouse antihuman CD38-FITC-conjugated antibody (Cytognos, Spain). Flow cytometry performed by FACS Calibur (BDFacs Calibur Becton Dickinson, USA). Results The total lymphocyte percentage was higher in PMN patients receiving either standard treatment or rituximab than in healthy controls (standard-treatment vs. control: P value = 0.001, pre-rituximab vs. control: P value =0.001). In post-rituximab analysis, CD19+ cell count showed notable reduction (P value = 0.003) B CD19+CD24+CD38- cells, representing the memory B cell population, did not show any significant difference between healthy controls and patients. Furthermore, the count of these cells did not decrease significantly two months after rituximab administration. The subset of CD19+CD24-CD38+ B lymphocytes, a class of naïve/mature lymphocytes with normal function, was significantly higher in the control group than in standard treatment patients (P value = 0.01). However, no statistically significant difference was found in CD19+CD24-CD38+B lymphocytes neither between the rituximab and control groups nor between pre-rituximab and post-rituximab patients. Conclusion The number of regulatory B cells decreased in both standard treatment and rituximab-receiving PMN patients and the proportion of naïve/mature B-lymphocytes was lower in the former group. Moreover, the memory B cells count did not reduce significantly two months after rituximab administration. Hence, it might be the best choice to target the memory B cell subset in immunosuppressive therapy while avoiding the Breg or naïve/mature B lymphocyte depletion to obtain more favorable sustained outcomes.

scholarly journals Plethysmographic and Biochemical Markers in the Diagnosis of Endothelial Dysfunction in Pediatric Acute Lymphoblastic Leukemia Survivors – New Applications

2018 ◽  
pp. 903-909 ◽  
Author(s):  
A. MASOPUSTOVÁ ◽  
P. JEHLIČKA ◽  
M. HUML ◽  
T. VOTAVA ◽  
L. TREFIL ◽  
...  
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Endothelial Dysfunction ◽  
Successful Treatment ◽  
Biochemical Markers ◽  
Lymphoblastic Leukemia ◽  
Reactive Hyperemia ◽  
Plasma Concentrations ◽  
Control Group ◽  
Healthy Controls ◽  
New Applications

Acute lymphoblastic leukemia (ALL) and its treatment are associated with endothelial dysfunction (ED) and increased cardiovascular risk in adulthood. There are no data on ED in children after successful treatment of ALL. We aimed to assess new ED in these children using the plethysmographic reactive hyperemia index (RHI) and biomarkers that are known to be related to ED. In all, 22 children (mean 15.6 years), after successful treatment of ALL, and 18 healthy subjects were included in this prospective study. RHI, plasma concentrations of asymmetric dimethyl arginine (ADMA), high-sensitive CRP (hsCRP) and E-selectin were measured in all children. RHI values were significantly lower in ALL patients when compared with healthy controls (p<0.05). hsCRP was significantly increased in ALL patients compared with the control group (p<0.001). E-selectin plasma levels were higher in ALL patients as compared to healthy controls (p=0.05). This is the first study that combines both plethysmographic and biochemical methods to assess ED in ALL survivors. Significantly decreased RHI with elevated plasma concentrations of biochemical markers imply a possible association with premature ED in ALL patients. The combined diagnostic approach seems to be a valuable tool for more accurate detection of ED and preventive cardiovascular management in these patients.

scholarly journals Annexin V Levels as an Indicator of Myocardial Cell Death after Myocardial Infarction

2020 ◽  
Vol 19 (2) ◽  
pp. 78-84
Author(s):  
Mohammed Karem Ahmed ◽  
◽  
Adil Hassan Mohammed ◽  
Amed Medb Athab
Keyword(s):  
Myocardial Infarction ◽  
Cell Death ◽  
Apoptotic Cell ◽  
Plasma Concentrations ◽  
Annexin V ◽  
Myocardial Cell ◽  
Early Morning ◽  
Control Group ◽  
P Value ◽  
The Mean

Background:Myocardial ischemia is associated with apoptosis of cardiomyocyte and because of apoptotic cell death is characterized by externalization of Phosphatidylserine on the cell membrane, so it is amenable to targeting by Annexin V. Objective: To compare plasma concentrations of Annexin V in patients who had an early infarct with patients without infarction. And to analyze the plasma concentration of Annexin V in relationship to cardiovascular risk factors. Patients and Methods: A Case-control study of 100 patients (case) who are diagnosed with Myocardial Infarction (MI) and admitted to the coronary care unit of Baqubah Teaching Hospital and another 100 patients homogenous in terms of age and gender and who attended the hospital for other cause than myocardial infarction is selected as the control group during a period between the first of April and the first of July 2019. A special questionnaire used to collect the required information, an early morning blood sample is taken to measure the level of Annexin V by ELISA, Student’s t-test, ANOVA test and Chi_square test to find an association and differences between variables. Results: The results showed that The mean Annexin V level is significantly higher in cases (10.48155ng/ml) than control (1.28803ng/ml) with p-value =0.001 and a sample taken within 24 hours after MI is significantly higher in the mean level of Annexin V then the sample taken after 24 hours of MI. Conclusion: Generally, the measurement of Annexin V level has provided a good diagnostic test to evaluate myocardial infarction. Keywords: Myocardial infarction, Annexin V, Phosphatidylserine, Apoptosis

FUNCTIONAL LIMITATIONS AND ADIPOKINES IN HIV-INFECTED OLDER ADULTS

2015 ◽  
pp. 1-6
Author(s):  
K.N. SHAH ◽  
Z. MAJEED ◽  
H. YANG ◽  
J.J. GUIDO ◽  
T.N. HILTON ◽  
...  
Keyword(s):  
Older Adults ◽  
Physical Function ◽  
Functional Limitations ◽  
Functional Decline ◽  
Plasma Concentrations ◽  
Community Dwelling ◽  
Control Group ◽  
Healthy Controls ◽  
Prospective Longitudinal ◽  
The Relationship

Background: There is a significant increase in the number of HIV-infected older adults (HOA). This population may experience functional decline at a much younger age. Little is known about the relationship between functional limitations and systemic adipokines in HOA. Objective: Our study aimed to evaluate the relationship between functional limitations and systemic adipokine levels in HOA population. Design: Cross-sectional. Setting: Academic hospital-based infectious disease clinic. Participants: The study investigated community-dwelling HIV-infected adults >50 years old and compared this group with age, gender and BMI comparable healthy controls. Measurements: We measured functional status, body composition and plasma concentrations of adipokines. Results: Fifty-four HOA were studied (mean: age 57 years, BMI 29 kg/m2, CD4 604, duration of HIV 17 years) and compared with thirty-two age, gender and BMI comparable healthy controls. The HOA group showed significantly higher functional limitations compared to the age, gender and BMI comparable controls (p<0.05). Levels of adipokines were significantly different between the two groups (p<0.05). Multiple regression analyses indicated that adiponectin and visfatin were significantly correlated with several physical function measures after controlling for age, sex, and metabolic comorbidities. Adiponectin was negatively correlated with functional limitations, and this relationship was stronger in the control group compared to the HOA group. Conversely, visfatin was positively correlated with functional limitations only in the HOA group. Conclusion: HOA have significant functional limitations and alteration in adipokine levels compared to controls. Adiponectin and visfatin were associated with functional limitations. Visfatin was a correlate of physical function only in the HOA group. Prospective longitudinal studies could provide further insight on the role of adipokines in HIV-related functional decline.

scholarly journals Evaluating the Neuroprotective Effect of Melatonin on Patients with Hemorrhagic Stroke Using Serum S100B Protein as a Prognostic Marker

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Hamidreza Sharifnia ◽  
Mojtaba Mojtahedzadeh ◽  
Mehrnoush Dianatkhah ◽  
Atabak Najafi ◽  
Arezoo Ahmadi ◽  
...  
Keyword(s):  
Mechanical Ventilation ◽  
Hemorrhagic Stroke ◽  
Neuroprotective Effect ◽  
Plasma Concentrations ◽  
Control Group ◽  
P Value ◽  
Neuroprotective Agents ◽  
Icu Stay ◽  
Baseline Values ◽  
Length Of Icu Stay

Background: Intracerebral hemorrhage (ICH) is one of the most debilitating kinds of stroke. Recent evidence shows that the proper initiation of neuroprotective agents might save at risk neurons and improve the outcome. Objectives: The focus of this study is to evaluate the neuroprotective effect of melatonin on patients with hemorrhagic stroke. Methods: Forty adult patients with confirmed nontraumatic ICH, who were admitted to the ICU within 24 hours of the stroke onset were enrolled in this study. Subjects in the melatonin group received 30 mg of melatonin every night for 5 consecutive nights. In order to evaluate the intensity of the neuronal injury, S100B was assessed once on day 1 and, day 5 post ICU admission. Additionally, the length of ICU stay, mortality, and the duration of mechanical ventilation were also recorded. Results: Forty patients completed the study. In both groups the plasma concentrations of S100B decreased after 5 days compared with their baseline values. However, this reduction was more significant in the melatonin compared to the control group (P-value < 0.05). The duration of mechanical ventilation and length of ICU stay was shorter in the melatonin group, and this difference was statistically significant for the length of ICU stay (P-value < 0.05), and marginally significant for the duration of mechanical ventilation (P-value = 0.065). The in-ICU mortality rate of the melatonin group was 15%, almost half of that of the control group (30%). However, this difference was not statistically significant. Conclusions: In conclusion, melatonin can be considered as a harmless and effective nueroprotective agent with some unique features which has made it an appropriate adjunctive medicine for critically ill intubated patients.

scholarly journals Association of 45-bp ins/del polymorphism of uncoupling protein 2 (UCP2) and susceptibility to nonalcoholic fatty liver and type 2 diabetes mellitus in North-west of Iran

2020 ◽  
Author(s):  
Salehe Rezapour ◽  
Shiva Ahdi Khosroshahi ◽  
Hadi Farajnia ◽  
Fatemeh Mohseni ◽  
Manouchehr Khoshbaten ◽  
...  
Keyword(s):  
Energy Homeostasis ◽  
Uncoupling Protein ◽  
White Blood Cells ◽  
Control Group ◽  
P Value ◽  
Uncoupling Protein 2 ◽  
Genotype Distribution ◽  
Healthy Controls ◽  
Nonalcoholic Fatty Liver ◽  
North West

Abstract Objective: Uncoupling protein 2 (UCP2) is a regulator of insulin secretion, free fatty acid (FFA) concentrations, and lipid metabolism that plays crucial roles in energy homeostasis. This study aimed to investigate the association of 45-bp ins/del polymorphism of UCP2 and susceptibility to NAFLD and T2DM. DNA was extracted from the white blood cells of the subjects and the gene polymorphism was determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In this study, 72 patients with NAFLD, 71 healthy individuals as control, 80 patients with T2DM, and 77 age/race matched healthy controls were enrolled in the study.Results: A higher prevalence of insertion/insertion genotype has been observed in T2DM patients compared with the control group (p value˂ 0.05). But, there was no difference in genotype distribution between NAFLD patients and control groups (p value> 0.05). NAFLD patients with D/D, D/I genotype had higher triglyceride, ALT and AST levels, and lower HDL level than healthy controls. Patients with T2DM with D/D or D/I genotype also had significantly higher fasting serum glucose (FSG) level. While we found an association between the 45bp I/D polymorphism in 3ʹUTR of UCP2 and T2DM, no any correlation between this polymorphism and NAFLD was identified.

Estimation Of Salivary Superoxide Dismutase Level In Oral Submucous Fibrosis: A Clinical And Biochemical Study

2019 ◽  
Vol 09 (02) ◽  
pp. 86-90
Author(s):  
Humera Akhlaq ◽  
Muhammad Owais Ismail ◽  
Muhammad Abdul Samad
Keyword(s):  
Superoxide Dismutase ◽  
Study Design ◽  
Oral Submucous Fibrosis ◽  
Mouth Opening ◽  
University Hospital ◽  
Ethical Review ◽  
Control Group ◽  
P Value ◽  
Strong Positive Correlation ◽  
Healthy Controls

Objective: The objective of the study was to compare the salivary Superoxide dismutase (mU/L) level among stage one patients of Oral sub mucous fibrosis and healthy controls. Study Design and Setting: It was a comparative cross sectional study design conducted at Outpatient clinic of Dental Department at Ziauddin University Hospital from January 2012 till December 2013. Methodology: Eighty histo-pathologically confirmed patients of clinical stage one Oral Sub mucosal fibrosis (OSMF) and eighty healthy controls were matched for age and gender to recruit in the study. Socio demographic information (i.e. age, gender, ethnicity, education and occupation), oral health status, measurement of mouth opening (mm) and Superoxide dismutase (mU/L) were recorded. The study was conducted after the approval granted from the ethical review committee of Ziauddin University Hospital. Data was entered and analyzed using SPSS version 21 (IBM). Result: The standard mean (+/-SD) of Superoxide dismutase (mU/L) in both groups was 4.99 (+/-2.18). The mean of Superoxide dismutase (mU/L) in confirmed cases of oral sub mucosal fibrosis was 3.02 (+/-0.44) which was significantly lower as compared to the control group which was 6.96(+/-1.24) and the calculated p value was 0.001.The strong positive correlation was identified in mouth opening (mm) and superoxide dismutase (0.842). Conclusion: It was concluded that the levels of salivary superoxide dismutase was significantly lower in the sample of OSMF and lower levels of superoxide dismutase (mU/L) was the indication of decrease in mouth opening among the cases of OSMF. Therefore, salivary superoxide dismutase can be used as a biomarker for the early detection as well as successful treatment of OSMF

Plasma D Dimer: A Useful Marker of Fibrin Breakdown in Renal Failure

1989 ◽  
Vol 61 (03) ◽  
pp. 522-525 ◽  
Author(s):  
M P Gordge ◽  
R W Faint ◽  
P B Rylance ◽  
H Ireland ◽  
D A Lane ◽  
...  
Keyword(s):  
Renal Failure ◽  
Renal Function ◽  
Acute Renal Failure ◽  
Monoclonal Antibodies ◽  
Diabetic Nephropathy ◽  
Renal Disease ◽  
Plasma Concentrations ◽  
Significant Negative Correlation ◽  
Healthy Controls ◽  
D Dimer

SummaryD dimer and other large fragments produced during the breakdown of crosslinked fibrin may be measured by enzyme immunoassay using monoclonal antibodies. In 91 patients with renal disease and varying degrees of renal dysfunction, plasma D dimer showed no correlation with renal function, whereas FgE antigen, a fibrinogen derivative which is known to be cleared in part by the kidney, showed a significant negative correlation with creatinine clearance. Plasma concentrations of D dimer were, however, increased in patients with chronic renal failure (244 ± 3l ng/ml) (mean ± SEM) and diabetic nephropathy (308 ± 74 ng/ml), when compared with healthy controls (96 ± 13 ng/ml), and grossly elevated in patients with acute renal failure (2,451 ± 1,007 ng/ml). The results indicate an increase in fibrin formation and lysis, and not simply reduced elimination of D dimer by the kidneys, and are further evidence of activated coagulation in renal disease. D dimer appears to be a useful marker of fibrin breakdown in renal failure.

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