Estimation of phosphorylation level of amyloid-beta isolated from human blood plasma: Ultrahigh-resolution mass spectrometry

Recently, amyloid-beta (Aβ) phosphorylation at position 8 has been shown to be associated with pathogenesis of Alzheimer’s disease. Since the modification occurs in the key fragment of the metal-binding domain of Aβ and should seriously affect the interaction of pS8-Aβ with zinc ions, this isoform may be a potential precursor of pathogenic oligomer forms of Aβ. Hence the level of pS8-Aβ in human biological fluids (blood, urine, cerebrospinal fluid) may reflect various stages of pathogenesis of the Alzheimer’s disease. The aim of the work was to develop a prototype of an analytical method for quantitative determination of the level of pS8-Aβ isoform in binary mixtures with native Aβ in order to further use it to estimate the levels of phosphorylated amyloid-beta in blood plasma samples of patients diagnosed with Alzheimer’s disease.     

Genotype Distribution,Viral Load and Clinical Characteristics of Infants with Postnatal or Congenital Cytomegalovirus Infection

Background

Congenital cytomegalovirus infection is a leading cause of long-term sequelae. Cytomegalovirus is also frequently transmitted to preterm infants postnatally, but these infections are mostly asymptomatic. A correlation between cytomegalovirus genotypes and clinical manifestations has been reported previously in infants with congenital infection, but not in preterm infants with postnatal infection.

Objectives

The main objective of this study was to investigate cytomegalovirus genotype distribution in postnatal and congenital cytomegalovirus infection and its association with disease severity.

Methods

Infants admitted to the neonatal intensive care unit of the University Medical Center Utrecht, The Netherlands between 2003–2010 and diagnosed with postnatal or congenital cytomegalovirus infection were included. Classification of cytomegalovirus isolates in genotypes was performed upon amplification and sequencing of the cytomegalovirus UL55 (gB) and UL144 genes. Clinical data, cerebral abnormalities, neurodevelopmental outcome and viral load were studied in relation to genotype distribution.

Results

Genotyping results were obtained from 58 preterm infants with postnatal cytomegalovirus infection and 13 infants with congenital cytomegalovirus infection. Postnatal disease was mild in all preterm infants and all had favourable outcome. Infants with congenital infection were significantly more severely affected than infants with postnatal infection. Seventy-seven percent of these infants were symptomatic at birth, 2/13 died and 3/13 developed long-term sequelae (median follow-up 6 (range 2–8) years). The distribution of cytomegalovirus genotypes was comparable for postnatal and congenital infection. UL55 genotype 1 and UL144 genotype 3 were predominant genotypes in both groups.

Conclusions

Distribution of UL55 and UL144 genotypes was similar in asymptomatic postnatal and severe congenital CMV infection suggesting that other factors rather than cytomegalovirus UL55 and UL144 genotype are responsible for the development of severe disease.     

Mining the human autoantibody repertoire: Isolation of potent IL17A-neutralizing monoclonal antibodies from a patient with thymoma

Anti-cytokine autoantibodies have been widely reported to be present in human plasma, both in healthy subjects and in patients with underlying autoimmune conditions, such as autoimmune polyendocrinopathy candidiasis ectodermal dystrophy (APECED) or thymic epithelial neoplasms. While often asymptomatic, they can cause or facilitate a wide range of diseases including opportunistic infections. The potential therapeutic value of specific neutralizing anti-cytokine autoantibodies has not been thoroughly investigated. Here we used mammalian cell display to isolate IL17A-specific antibodies from a thymoma patient with proven high-titer autoantibodies against the same. We identified 3 distinct clonotypes that efficiently neutralized IL17A in a cell-based in vitro assay. Their potencies were comparable to those of known neutralizing antibodies, including 2, AIN457 (secukinumab) and ixekizumab that are currently in clinical development for the treatment of various inflammatory disorders. These data clearly demonstrate that the human autoantibody repertoire can be mined for antibodies with high therapeutic potential for clinical development.     

SARS-CoV-2 Nsp3 unique domain SUD interacts with guanine quadruplexes and G4-ligands inhibit this interaction

The multidomain non-structural protein 3 (Nsp3) is the largest protein encoded by coronavirus (CoV) genomes and several regions of this protein are essential for viral replication. Of note, SARS-CoV Nsp3 contains a SARS-Unique Domain (SUD), which can bind Guanine-rich non-canonical nucleic acid structures called G-quadruplexes (G4) and is essential for SARS-CoV replication. We show herein that the SARS-CoV-2 Nsp3 protein also contains a SUD domain that interacts with G4s. Indeed, interactions between SUD proteins and both DNA and RNA G4s were evidenced by G4 pull-down, Surface Plasmon Resonance and Homogenous Time Resolved Fluorescence. These interactions can be disrupted by mutations that prevent oligonucleotides from folding into G4 structures and, interestingly, by molecules known as specific ligands of these G4s. Structural models for these interactions are proposed and reveal significant differences with the crystallographic and modeled 3D structures of the SARS-CoV SUD-NM/G4 interaction. Altogether, our results pave the way for further studies on the role of SUD/G4 interactions during SARS-CoV-2 replication and the use of inhibitors of these interactions as potential antiviral compounds.     

HABITAT USE AND PREFERENCES OF INDO-PACIFIC HUMPBACK DOLPHINS SOUSA CHINENSIS IN ALGOA BAY, SOUTH AFRICA

This paper examines environmental and behavioral determinants of the habitat use and preferences of Indo-Pacific humpback dolphins inhabiting the Algoa Bay region on the south Eastern Cape coast of South Africa. In order to quantify the habitat use and preference, two indices were used, the Coefficient of Area Use (AU) and the Activity Index (AI). The dolphins inhabit a narrow strip of shallow, inshore waters of Algoa Bay and remain mostly within 400 m of the shore, in water less than 15 m deep, with no apparent preference for clear or turbid water. Water depth is probably the main factor limiting their inshore distribution, and the 25-m isobath seems to represent the critical depth. Within this confined, inshore distribution, dolphin activities concentrate in the vicinity of rocky reefs-their primary feeding grounds. Dolphin dependence on these shallow-water habitats is evident throughout the year and, consequently, the inshore shallow reefs are identified as the "key habitat" which is of primary importance for humpback dolphins in Eastern Cape waters. The dolphins' dependence on this restricted type of habitat within an already restricted inshore distribution makes them particularly vulnerable to alteration or loss of this habitat.     

Structural studies and physiological activity of lemnan, a pectin fromLemna minor L

A pectic polysaccharide, lemnan, was isolated from freshly collected duckweedLemna minor L. Its sugar chain was shown to be mainly composed of the residues ofD-galacturonic acid (64%), galactose, arabinose, xylose, andD-apiose, a branched chain sugar. The high content ofD-apiose (25%) indicated that lemnan is an apiogalacturonan type pectin similar to zosteran, a pectic polysaccharide from a sea phanerogam of the Zosteraceae family. The results of partial acidic hydrolysis, pectinase digestion, and NMR studies of lemnan demonstrated that its macromolecule contains regions of the linear α-1,4-D-galacturonan and branched apiogalacturonan. The side chains of apiogalacturonan were found to be formed of single and 1,5-linked residues ofD-apiofuranose attached to 2- and 3-positions of theD-galacturonic acid residues of the apiogalacturonan backbone. Lemnan was shown to exhibit an immunomodulatory effect activating the system of phagocytosis.     

Alterations in Nitric Oxide Activity and Sensitivity in Early Streptozotocin-Induced Diabetes Depend on Arteriolar Size

Changes in NO activity may play an important role in the early increase in microvascular flow that has been implicated in the pathogenesis of diabetic microangiopathy. We assessed, in the in situ spinotrapezius muscle preparation of 6 weeks'' streptozotocin-diabetic rats (n = 6) and of agematched controls (n = 8), basal inside diameters of A2–A4 arterioles and the reactivity to topically applied acetylcholine and nitroprusside, before and after N^G-nitro-L-arginine. In diabetic rats, cholinergic vasodilatation in A2–A4 arterioles was intact. Basal diameter in A3 and A4 arterioles was significantly higher in streptozotocin-diabetic rats. The increased basal diameter in A3 arterioles was partially due to an increased contribution of NO to basal diameter. The response to nitroprusside was impaired in streptozotocin-diabetic rats in A2, but not in A3 and A4 arterioles. Thus, this study shows that NO activity and sensitivity are altered after 6 weeks of streptozotocin-induced diabetes. These streptozotocin-induced changes are anatomically specific and, for arterioles, depend on their position within the vascular tree.