The Role of Plasminogen Activator Inhibitor Type 1 (PAI-1) in Placenta-Mediated Pregnancy Complications: A Systematic Review

Plasminogen activator inhibitor type 1 (PAI-1) is a main inhibitor of fibrinolysis. The PAI-1 gene (SERPINE1) harbors genetic variants with the potential of modifying plasma levels of PAI-1. A delicate balance exists between the coagulation and fibrinolytic system, and changes in PAI-1 have been suggested to compromise establishment of a successful pregnancy. Therefore, this systematic review investigated the association between genetic variants and/or plasma levels of PAI-1 and placenta-mediated pregnancy complications. An extensive literature search was conducted in PubMed, Embase, and Web of Science on the 29th of April 2021. All studies underwent quality rating according to The Study Quality Assessment Tools checklist provided by National Heart, Lung and Blood Institute. A total of 71 studies were included, among which 60 studies investigated PAI-1 genotypes and 11 studies measured PAI-1 plasma levels. In 32 out of 59 studies, no association was found between the PAI-1 4G/5G polymorphism (rs1799768) and placenta-mediated pregnancy complications, which was stated as no significant difference in the genotype distribution comparing women with and without placenta-mediated pregnancy complications or no significantly increased odds of placenta-mediated pregnancy complications carrying the 4G/4G or 4G/5G genotype. Eight out of 11 studies reported significantly higher PAI-1 plasma levels in preeclamptic women than in women without preeclampsia. In conclusion, no clear evidence indicates that PAI-1 polymorphisms are associated with placenta-mediated pregnancy complications, and the possible association between high PAI-1 plasma levels and preeclampsia needs further investigations. Thus, investigation of PAI-1 genotypes and PAI-1 plasma levels does not currently seem to have a place in daily clinical practice managing placenta-mediated pregnancy complications.

Analytical Techniques for the Analysis of Lopinavir and Ritonavir in Pharmaceutical Dosage Form and Biological Matrices: A Review

Background: Lopinavir and Ritonavir are the protease inhibitor type of anti-retroviral drugs. Both are used for the treatment of HIV/AIDS. This paper reviews many analytical methods for the analysis of LPV and RTV in pharmaceutical formulations (tablet, capsule, syrup, and bulk) and biological fluids (human plasma, serum, cerebrospinal fluid, rat plasma, and human hair). Objective: The study aims to summarize various ana¬lytical techniques, such as Chromatography, Spectrophotometry; and also hyphenated techniques, such as LC-MS/MS, UPLC-MS for analysis of Lopinavir and Ritonavir. Method: The review deals with com¬prehensive details about the type of various analytical techniques, such as spectroscopy (UV), chromatography (RP-HPLC, HPTLC, UPLC), and hyphenated techniques, i.e., LC-MS/MS, UPLC-MS for the analysis of lopinavir and ritonavir. These techniques are either explored for the quantification, de¬tection of metabolite or for stability studies of the LPV & RTV. Conclusion: The present studies revealed that the HPLC technique along with the spectro-scopic, have been most widely used for the analysis. Out of the developed methods, hyphenated UPLC-MS and LC-MS are very sensitive and helps in the easy estimation of drugs compared to that of the other techniques. This review may provide comprehensive details to the researchers working in the area of analytical research of LPV & RTV.

Design and Development of Autotaxin Inhibitors

Autotaxin (ATX) is the only enzyme of the ecto-nucleotide pyrophosphatase/phosphodiesterase (ENPP2) family with lysophospholipase D (lysoPLD) activity, which is mainly responsible for the hydrolysis of extracellular lysophosphatidylcholine (LPC) into lysophosphatidic acid (LPA). LPA can induce various responses, such as cell proliferation, migration, and cytokine production, through six G protein-coupled receptors (LPA1-6). This signaling pathway is associated with metabolic and inflammatory disorder, and inhibiting this pathway has a positive effect on the treatment of related diseases, while ATX, as an important role in the production of LPA, has been shown to be associated with the occurrence and metastasis of tumors, fibrosis and cardiovascular diseases. From mimics of ATX natural lipid substrates to the rational design of small molecule inhibitors, ATX inhibitors have made rapid progress in structural diversity and design over the past 20 years, and three drugs, GLPG1690, BBT-877, and BLD-0409, have entered clinical trials. In this paper, we will review the structure of ATX inhibitors from the perspective of the transformation of design ideas, discuss the advantages and disadvantages of each inhibitor type, and put forward prospects for the development of ATX inhibitors in the future.

Demonstration of three distinct high molecular weight complexes between plasminogen activator inhibitor type 1 and tissue type plasminogen activator.

Background: Details of the molecular interaction between tissue type plasminogen activator (tPA) and plasminogen activator inhibitor type-1 (PAI-1) remain unknown. Methods and Results: Three distinct forms of high molecular weight complexes are demonstrated. Two of the forms were detected by mass spectrometry. The high molecular mass detected by MALDI-TOF MS spectrometry was 107,029 Da, which corresponds to the sum of molecular masses of the intact tPA (65,320 Da) and the intact PAI-1 (42,416 Da). The lower molecular mass was 104,367 Da and is proposed to lack the C-terminal bait peptide of PAI-1 (calculated mass, 3,804 Da) which was detected as a 3,808 Da fragment. When the complex was analyzed by SDS-PAGE, only a single band was observed. However, after treatment by SDS and Triton X-100, two distinct forms of the complex with different mobilities were shown by SDS-PAGE. The higher molecular weight band demonstrated specific tPA activity on fibrin autography, whereas the lower molecular weight band did not. Peptide sequence analysis of these two bands, however, unexpectedly revealed the existence of the C-terminal cleavage peptide in both bands and its amount was less in the upper band. In the upper band, the sequences corresponding to the regions at the interface between two molecules in its Michaelis intermediate were diminished. Thus, these two bands corresponded to distinct nonacyl-enzyme complexes, wherein only the upper band liberated free tPA under the conditions employed. Conclusion: These data suggest that under physiological conditions a fraction of the tPA-PAI-1 population exists as non-acylated-enzyme inhibitor complex.

Case Report: Placental Maternal Vascular Malperfusion Affecting Late Fetal Development and Multiorgan Infection Caused by SARS-CoV-2 in Patient With PAI-1 4G/5G Polymorphism

Background: Pregnant women are susceptible to the novel coronavirus (SARS-CoV-2), and the consequences for the fetus are still uncertain. Here, we present a case of a pregnant woman with subclinical hypothyroidism and a plasminogen activator inhibitor type 1 (PAI-1) 4G/5G polymorphism who was infected with SARS-CoV-2 at the end of the third trimester of pregnancy, with unexpected evolution of death of the newborn 4 days postpartum.Methods: Nested PCR was performed to detect the virus, followed by ssDNA sequencing.Results: Transplacental transmission of SARS-CoV-2 can cause placental inflammation, ischemia, and neonatal viremia, with complications such as preterm labor and damage to the placental barrier in patients with PAI-1 4G/5G polymorphism.Conclusion: We showed a newborn with several damages potentially caused due to the PAI-1 polymorphisms carried by the mother infected with SARS-CoV-2 during pregnancy.

Effect of Storage Temperature and Postharvest Tuber Treatment with Chemical and Biorational Inhibitors on Suppression of Sprouts During Potato Storage

Consumption of ware potatoes in the tropics may be enhanced by storage under conditions that inhibit sprouting, most important of which is the temperature. The effects of storage temperatures (10 °C and 23 °C) and two alternatives to the chlorpropham (CIPC): 1,4-dimethylnaphthalene (DMN) and peppermint oil (PMO), a biorational inhibitor, were evaluated for tuber sprout suppression at postharvest. Tubers of three cultivars – ‘Asante’, ‘Kenya Mpya’, and ‘Shangi’ – with different dormancy lengths were assessed. Storage temperatures and suppressant effects on tuber sprouting, sprout length and sprout numbers per tuber were investigated in replicated storage experiments. Significant differences in sprout inhibition were observed between the two temperatures with suppressive effects higher at 10 °C than in 23 °C. CIPC had the greatest suppressive effect on tuber sprout inhibition at both temperatures. After 24 weeks of storage, the suppressive effects of CIPC at 10 °C was 100% on the three cultivars, whereas all the nontreated tubers sprouted after six (‘Shangi’), twelve (‘Asante’), fourteen (‘Kenya Mpya’) weeks of storage. At 23 °C, the effectiveness of CIPC followed the dormancy period of the cultivars, with ‘Shangi’ recording significantly more sprouted tubers followed by ‘Asante’ and ‘Kenya Mpya’. The inhibition of sprouting by DMN and PMO varied with storage temperatures and cultivars, but were significantly greater than the nontreated tubers. At 10 °C, the effectiveness of DMN treatment was equal to that of CIPC for ‘Kenya Mpya’ resulting in 100% inhibition. In the PMO treatments, complete inhibition of sprouting was observed for 18 weeks on cultivars ‘Asante’, ‘Kenya Mpya’ and 14 weeks on ‘Shangi’. At 23 °C, PMO suppressed sprouts for 14 weeks on ‘Asante’ and ‘Kenya Mpya’ and on ‘Shangi’ for 8 weeks. Storage temperature effects on sprout length and numbers varied with inhibitor type and cultivar. These findings showed that in potato tuber storage, inhibitors can replace low temperature but to an extent depending on the cultivar dormancy character and storage length, thus enabling greater consumption of potatoes in tropical regions.