Matrix metalloproteinas-9 functional promoter polymorphism 1562C>T increased risk of early-onset coronary artery disease

The Matrix metalloproteinas-9 functional promoter polymorphism 1562C>T may be considered an important genetic determinant of early-onset coronary artery disease (ECAD). In this study, association between MMP-9 1562C>T allele with plasma MMP-9 activity, homocysteine and lipid–lipoproteins level and ECAD in Iranian subjects was investigated. This case–control study consisted of 53 ECAD patients (age < 55 years) and unrelated late-onsets CAD (age > 70 years) who angiographically had at least 50% stenosis. MMP-9 1562C>T polymorphism was detected by PCRRFLP, plasma MMP-9 activity, serum lipid and homocysteine levels were determined by gelatin gel zymography, enzyme assay and by HPLC, respectively. The presence of MMP-9 1562C>T allele was found to be associated with ECAD (OR = 3.2, P = 0.001). The ECAD patients with MMP-9 1562C>T allele had higher MMP-9 activity (P = 0.001), LDL-C (P = 0.045), TC (P = 0.02) and homocysteine (P = 0.01) levels than the LCAD subjects. MMP-9 1562C>T allele is a risk factor for ECAD. The carriers of this allele have high levels of MMP-9 activity, LDL-C, TC and homocysteine (P = 0.01), thus, are more likely to develop myocardial infarction and CAD at young age (less than 55 years).     

Assignment of ring size in isopropylidene acetals by carbon-13 n.m.r. spectroscopy

A series of 1-O-acylaldose derivatives was prepared in good yield through the reaction of 1,3-dialkyl-O-glycosylpseudoureas with carboxylic acids.     

Purification and characterization of genetic variants of 6-phosphogluconate dehydrogenase

This paper describes the physicochemical characteristics in partially purified enzyme on subjects with the Pd A, Pd AB, and Pd B variants of 6-phosphogluconic dehydrogenase (6PGD). For these studies, whole blood was purified about 225-fold using ion exchange chromatography on DEAE cellulose column and fractionation with ammonium sulfate. 6PGD emerges as a single peak between 0.01 m and 0.1 m phosphate buffer on the column and is precipitated in the 55–80% fraction of ammonium sulfate. This purified enzyme can be stored frozen for several months without appreciable loss of activity and contains no detectable activity of glucose 6-phosphate dehydrogenase and glutathione reductase. The three variants of partially purified 6PGD varied from each other in two respects. The transitional temperature is 47.8 C for Pd A, 45.4 C for Pd AB, and 41.1 C for Pd B. The K _m for 6PGA is 30 m for Pd A, 21 m for Pd AB, and 15 mfor Pd B. These observations add further strength to the concept that the polymorphism in 6PGD represents alterations in either the configuration or structure of the protein molecule itself.Supported by grants from the Chicago Community Trust and the U.S. Public Health Service (Tl-AM-5186).     

Unusual susceptibility of heme proteins to damage by glucose during non-enzymatic glycation

Glucose modifies the amino groups of proteins by a process of non-enzymatic glycation, leading to potentially deleterious effects on structure and function that have been implicated in the pathogenesis of diabetic complications. These changes are extremely complex and occur very slowly. We demonstrate here that hemoglobin and myoglobin are extremely susceptible to damage by glucose in vitro through a process that leads to complete destruction of the essential heme group. This process appears in addition to the expected formation of so-called advanced glycation end products (AGEs) on lysine and other side-chains. AGE formation is enhanced by the iron released. In contrast, the heme group is not destroyed during glycation of cytochrome c, where the sixth coordination position of the heme iron is not accessible to solvent ligands. Glycation leads to reduction of ferricytochrome c in this case. Since hydrogen peroxide is known to destroy heme, and the destruction observed during glycation of hemoglobin and myoglobin is sensitive to catalase, we propose that the degradation process is initiated by hydrogen peroxide formation. Damage may then occur through reaction with superoxide generated (a reductant of ferricytochrome c), or hydroxyl radicals, or with both.     

Phytochemical and morphological characterization of Satureja khuzistanica Jamzad populations from Iran

Satureja khuzistanica is an endemic herb growing wild in Iran with interesting pharmacological and biological properties. Here, as an initial step of the domestication process, the variability of phytochemical and morphological traits among 69 individuals of eight natural populations of the plant was studied. The investigated characteristics were the essential oil content and composition, the rosmarinic acid (RA) content, and the leaf and flower morphologies. The Abdanan and Kaver populations showed the highest oil contents. The characterization by GC-FID and GC/MS analyses of the oils revealed that all 69 sampled individuals had carvacrol as the main component with very high contents (89.59-95.41%). The content of RA of the MeOH extracts of S. khuzistanica showed a high level of variability (coefficient of variation (CV) 50.0%) ranging from 0.59% (w/w) in the Paalam population to 1.81% (w/w) in the Abdanan population. The peduncle length and the leaf surface area (CVs of 47.39 and 47.21%, resp.) were the most variable morphological characteristics among the examined populations. The high level of phytochemical and morphological variability among the studied populations suggests a breeding approach during the domestication, to gain new, promising, and homogenous cultivars, attractive for the industry and agriculture.     

BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models

BioSig3D is a computational platform for high-content screening of three-dimensional (3D) cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony. BioSig3D also enables visualization of raw and processed 3D volumetric data for quality control, and integrates advanced bioinformatics analysis. The system consists of multiple computational and annotation modules that are coupled together with a strong use of controlled vocabularies to reduce ambiguities between different users. It is a web-based system that allows users to: design an experiment by defining experimental variables, upload a large set of volumetric images into the system, analyze and visualize the dataset, and either display computed indices as a heatmap, or phenotypic subtypes for heterogeneity analysis, or download computed indices for statistical analysis or integrative biology. BioSig3D has been used to profile baseline colony formations with two experiments: (i) morphogenesis of a panel of human mammary epithelial cell lines (HMEC), and (ii) heterogeneity in colony formation using an immortalized non-transformed cell line. These experiments reveal intrinsic growth properties of well-characterized cell lines that are routinely used for biological studies. BioSig3D is being released with seed datasets and video-based documentation.     

Buckyballs conjugated with nucleic acid sequences identifies microorganisms in live cell assays

Background

Rapid identification of bacteria can play an important role at the point of care, evaluating the health of the ecosystem, and discovering spatiotemporal distributions of a bacterial community. We introduce a method for rapid identification of bacteria in live cell assays based on cargo delivery of a nucleic acid sequence and demonstrate how a mixed culture can be differentiated using a simple microfluidic system.

Methods

C60 Buckyballs are functionalized with nucleic acid sequences and a fluorescent reporter to show that a diversity of microorganisms can be detected and identified in live cell assays. The nucleic acid complexes include an RNA detector, targeting a species-specific sequence in the 16S rRNA, and a complementary DNA with an attached fluorescent reporter. As a result, each bacterium can be detected and visualized at a specific emission frequency through fluorescence microscopy.

Results

The C60 probe complexes can detect and identify a diversity of microorganisms that include gram-position and negative bacteria, yeast, and fungi. More specifically, nucleic-acid probes are designed to identify mixed cultures of Bacillus subtilis and Streptococcus sanguinis, or Bacillus subtilis and Pseudomonas aeruginosa. The efficiency, cross talk, and accuracy for the C60 probe complexes are reported. Finally, to demonstrate that mixed cultures can be separated, a microfluidic system is designed that connects a single source-well to multiple sinks wells, where chemo-attractants are placed in the sink wells. The microfluidic system allows for differentiating a mixed culture.

Conclusions

The technology allows profiling of bacteria composition, at a very low cost, for field studies and point of care.

     

Neuraminidase-associated plasminogen recruitment enables systemic spread of natural avian Influenza viruses H3N1

Repeated outbreaks due to H3N1 low pathogenicity avian influenza viruses (LPAIV) in Belgium were associated with unusually high mortality in chicken in 2019. Those events caused considerable economic losses and prompted restriction measures normally implemented for eradicating high pathogenicity avian influenza viruses (HPAIV). Initial pathology investigations and infection studies suggested this virus to be able to replicate systemically, being very atypical for H3 LPAIV. Here, we investigate the pathogenesis of this H3N1 virus and propose a mechanism explaining its unusual systemic replication capability. By intravenous and intracerebral inoculation in chicken, we demonstrate systemic spread of this virus, extending to the central nervous system. Endoproteolytic viral hemagglutinin (HA) protein activation by either tissue-restricted serine peptidases or ubiquitous subtilisin-like proteases is the functional hallmark distinguishing (H5 or H7) LPAIV from HPAIV. However, luciferase reporter assays show that HA cleavage in case of the H3N1 strain in contrast to the HPAIV is not processed by intracellular proteases. Yet the H3N1 virus replicates efficiently in cell culture without trypsin, unlike LPAIVs. Moreover, this trypsin-independent virus replication is inhibited by 6-aminohexanoic acid, a plasmin inhibitor. Correspondingly, in silico analysis indicates that plasminogen is recruitable by the viral neuraminidase for proteolytic activation due to the loss of a strongly conserved N-glycosylation site at position 130. This mutation was shown responsible for plasminogen recruitment and neurovirulence of the mouse brain-passaged laboratory strain A/WSN/33 (H1N1). In conclusion, our findings provide good evidence in natural chicken strains for N1 neuraminidase-operated recruitment of plasminogen, enabling systemic replication leading to an unusual high pathogenicity phenotype. Such a gain of function in naturally occurring AIVs representing an established human influenza HA-subtype raises concerns over potential zoonotic threats.     

Evaluation of expression CXCL8 chemokine and its relationship with oocyte maturation and embryo quality in the intracytoplasmic sperm injection method

Molecular Biology Reports - The present study aimed to evaluate the expression of the chemokine CXCL8 in both mRNA and protein levels in the serum, follicular fluid (FF), and cumulus cells (CCs)...