Ultrastructural,cytochemical, and enzymatic studies on the adhesive “plaques” of the brown algaeLaminaria saccharina (L.) lamour. andNereocystis luetkeana (nert.) post. et rupr. 1

Summary Newly settled zoospores of bothLaminaria saccharina andNereocystis luetkeana are surrounded by adhesive plaques. At this stage cell wall deposition has not occurred and similar structures cannot be detected in the cytoplasm. In free swimming zoospores, however, plaques are observed exclusively in small vesicles within the cytoplasm. Enzymatic extractions and cytochemical tests indicate that plaques are glycoproteid in nature. Studies on the influence of several enzymes on the attachment of zoospores show that plaques are involved in the adhesion of zoospores to the substratum.Project supported by NRC grant #A2288. Nereocystis data are part of a Ph.D. dissertation presented by this author to the University of British Columbia.     

Interferon-induced tetherin restricts vesicular stomatitis virus release in neurons

Tetherin, a recently identified interferon (IFN)-inducible, type 2 transmembrane protein, has been shown to be a cellular antiviral restriction factor that retains newly formed virions in infected cells. Thus, tetherin plays an important role in the innate cell-autonomous immune response. The aim of this study was to examine the antiviral activities of tetherin in vesicular stomatitis virus infections of murine neuronal cells. Both IFN-β and IFN-γ induce the expression of tetherin mRNA and protein. Tetherin knockdown experiments were carried out by transfection of tethrin shRNA into murine neuroblastoma cells using a vector containing the pCMV-driven tGFP gene. The efficiency of transfection was monitored through GFP expression by the transfected cells. Selected transfected cells were used for further mRNA and protein analysis, fluorescent immunocytolocalization, and viral infection to study the impact of tetherin knockdown. Our research indicates that tetherin is expressed on the outer face of the plasma membrane of murine neuroblastoma cells, its expression can be induced with both IFN-γ and IFN-β, and tetherin restricts progeny virus release up to 100-fold in mammalian neurons, thus contributing to a potent antiviral state within the host cell.     

Comparison between blood hemoglobin concentration determined by point-of-care device and complete blood count in adult patients with dengue

BackgroundHematocrit measurement has been an indispensable tool for monitoring plasma leakage and bleeding in dengue patients. However, hematocrit measurement by automated methods is hampered by frequent venipunctures. Utility of point-of-care hemoglobin (POC-Hb) test for monitoring dengue patients has not been established. We evaluated the relationship between hemoglobin measured by POC-Hb testing and hematocrit measured by the automated method in adult dengue patients.Methodology and principal findingsAdult dengue patients were recruited at two university hospitals in Thailand from October 2019 to December 2020. POC-Hb test was performed using capillary blood simultaneously with venipuncture to obtain whole blood for an automated complete blood count (CBC) analysis. The correlation of hemoglobin and hematocrit measurement was evaluated. A total of 44 dengue patients were enrolled. Twenty-nine patients (65.9%) were female, with a median age of 31 years (interquartile range 22–41). Of the enrolled patients, 30 (68.2%), 11 (25.0%), and 3 (6.8%) were classified as dengue without warning signs, with warning signs, and severe dengue, respectively. Seven patients (15.9%) had hemoconcentration, and five patients (11.3%) had bleeding. A total of 216 pairs of POC-Hb and CBC were evaluated. A significant positive correlation was observed between hemoglobin measured by POC-Hb testing and hematocrit measured by an automated CBC (r = 0.869, p <0.001). Bland-Altman analysis between hemoglobin measured by POC-Hb testing and an automated CBC showed a bias of -0.43 (95% limit of agreement of -1.81 and 0.95). Using the cutoff of POC-Hb ≥20% as a criteria for hemoconcentration, the sensitivity and specificity of hemoconcentration detected by POC-Hb device were 71.4% and 100.0%, respectively.ConclusionsHemoglobin measurement by POC-Hb testing has a strong correlation with hematocrit in adult patients with dengue fever. However, the sensitivity in detecting hemoconcentration is fair. The adjunct use of capillary POC-Hb testing can decrease the frequency of venipuncture. Further study in children is encouraged.     

Simulations of DNA topoisomerase 1B bound to supercoiled DNA reveal changes in the flexibility pattern of the enzyme and a secondary protein–DNA binding site

Human topoisomerase 1B has been simulated covalently bound to a negatively supercoiled DNA minicircle, and its behavior compared to the enzyme bound to a simple linear DNA duplex. The presence of the more realistic supercoiled substrate facilitates the formation of larger number of protein–DNA interactions when compared to a simple linear duplex fragment. The number of protein–DNA hydrogen bonds doubles in proximity to the active site, affecting all of the residues in the catalytic pentad. The clamp over the DNA, characterized by the salt bridge between Lys369 and Glu497, undergoes reduced fluctuations when bound to the supercoiled minicircle. The linker domain of the enzyme, which is implicated in the controlled relaxation of superhelical stress, also displays an increased number of contacts with the minicircle compared to linear DNA. Finally, the more complex topology of the supercoiled DNA minicircle gives rise to a secondary DNA binding site involving four residues located on subdomain III. The simulation trajectories reveal significant changes in the interactions between the enzyme and the DNA for the more complex DNA topology, which are consistent with the experimental observation that the protein has a preference for binding to supercoiled DNA.     

The morphology of a harpacticoid copepod gut: A review and synthesis

The digestive tract of a harpacticoid copepod, Tigriopus californicus (Baker), was studied by using techniques of light and electron microscopy. Four cell types could be distinguished: type 1, an embryonic cell which will replace cells worn away or lost during secretion; type 2, a cell which synthesizes and secretes proteins and also plays a role in lipid absorption; and types 3 and 4, two cell types which absorb lipids. From the abundance of each cell type, the length of microvilli, the development of basal plasma membrane (PM), and luminal projections, the following conclusions were made. (1) The midgut caecum absorbs digested nutrients. (2) The anterior midgut absorbs nutrients and more importantly functions in merocrine and exocrine secretion. The presence of concretions in cell types 2 and 3 in the anterior midgut suggests that these tissues contribute in excretion, and in water and/or ion regulation. (3) The posterior midgut absorbs nutrients and contributes some holocrine secretion.     

Mitochondrial genome SNPs analysis of eight barley genotypes displaying hot spot regions,phylogenetic relationships and heteroplasmy

Abstract

Organellar genomes are small, circular entities that provide unique advantages as compared to the nuclear genome. The present study was aimed at evaluating the efficiency of utilizing mitochondrial single nucleotide polymorphisms (SNPs) approach in separating barley cultivars. Sequences generated via next-generation sequencing were further utilized to confirm the incidence of heteroplasmy in barley mitochondrial genome. The analysis involved seven cultivated barley (Hordeum vulgare subsp. vulgare) (VG) and one wild (H. vulgare subsp. spontaneum) (SP) genotypes. A total of 73 million paired-end reads per mitochondrial genomes across the eight barley genotypes were generated using Illumina HiSeq 2000 platform. Sequences of each genotype were separately aligned to the published barley mitochondrial reference genome, thus SNPs were detected. The overall results indicated the efficiency of using mitochondrial SNPs as a molecular marker in distinguishing among barley genotypes. Unique SNPs were determined in six out of the eight genotypes, where Giza131 and Giza129 had no specific mitochondrial SNPs, while Giza130 showed the largest number of unique mitochondrial SNPs. The phylogenetic tree indicated the close relationship between Giza129 and Giza130. Interestingly, SP was not clearly discriminated among genotypes.