PIVL,a new serine protease inhibitor from Macrovipera lebetina transmediterranea venom,impairs motility of human glioblastoma cells

A novel Kunitz-type serine proteinase inhibitor, termed PIVL, was purified to homogeneity from the venom of the Tunisian snake Macrovipera lebetina transmediterranea. It is a monomeric polypeptide chain cross-linked by three disulfide linkages with an isotope-averaged molecular mass of 7691.7 Da. The 67-residue full-length PIVL sequence was deduced from a venom gland cDNA clone. Structurally, PIVL is built by a single Kunitz/BPTI-like domain. Functionally, it is able to specifically inhibit trypsin activity. Interestingly, PIVL exhibits an anti-tumor effect and displays integrin inhibitory activity without being cytotoxic. Here we show that PIVL is able to dose-dependently inhibit the adhesion, migration and invasion of human glioblastoma U87 cells. Our results also show that PIVL impairs the function of αvβ3 and to a lesser extent, the activity of αvβ6, αvβ5, α1β1 and α5β1 integrins. Interestingly, we demonstrate that the ⁴¹RGN⁴³ motif of PIVL is likely responsible for its anti-cancer effect. By using time lapse videomicroscopy, we found that PIVL significantly reduced U87 cells motility and affected cell directionality persistence by 68%. These findings reveal novel pharmacological effects for a Kunitz-type serine proteinase inhibitor.     

Genome-wide regression models considering general and specific combining ability predict hybrid performance in oilseed rape with similar accuracy regardless of trait architecture

Key Message

Genomic prediction using the Brassica 60 k genotyping array is efficient in oilseed rape hybrids. Prediction accuracy is more dependent on trait complexity than on the prediction model.

Abstract

In oilseed rape breeding programs, performance prediction of parental combinations is of fundamental importance. Due to the phenomenon of heterosis, per se performance is not a reliable indicator for F₁-hybrid performance, and selection of well-paired parents requires the testing of large quantities of hybrid combinations in extensive field trials. However, the number of potential hybrids, in general, dramatically exceeds breeding capacity and budget. Integration of genomic selection (GS) could substantially increase the number of potential combinations that can be evaluated. GS models can be used to predict the performance of untested individuals based only on their genotypic profiles, using marker effects previously predicted in a training population. This allows for a preselection of promising genotypes, enabling a more efficient allocation of resources. In this study, we evaluated the usefulness of the Illumina Brassica 60 k SNP array for genomic prediction and compared three alternative approaches based on a homoscedastic ridge regression BLUP and three Bayesian prediction models that considered general and specific combining ability (GCA and SCA, respectively). A total of 448 hybrids were produced in a commercial breeding program from unbalanced crosses between 220 paternal doubled haploid lines and five male-sterile testers. Predictive ability was evaluated for seven agronomic traits. We demonstrate that the Brassica 60 k genotyping array is an adequate and highly valuable platform to implement genomic prediction of hybrid performance in oilseed rape. Furthermore, we present first insights into the application of established statistical models for prediction of important agronomical traits with contrasting patterns of polygenic control.

     

Molecular Genotyping of <Emphasis Type="Italic">Candida parapsilosis</Emphasis> Species Complex

Background

The Candida parapsilosis complex species has emerged as an important cause of human disease. The molecular identification of C. parapsilosis isolates at the species level can be helpful for epidemiological studies and then for the establishment of appropriate therapies and prophylactic measures.

Methods

The present study was undertaken to analyze 13 short tandem repeat (STR) markers (7 minisatellites and 6 microsatellites) in a global set of 182 C. parapsilosis complex isolates from different origins including invasive and superficial clinical sites.

Results

Upon the analysis of 182 strains of C. parapsilosis complex species, 10–17 haplotypes were detected for each minisatellite marker. The combination of 7 minisatellite markers yielded 121 different genotypes with a 0.995 D value. Upon the analysis of 114 isolates (68 from invasive infections and 46 from superficial infections), 21–32 genotypes were detected for each microsatellite marker. The combination of all 13 markers yielded 96 different genotypes among 114 isolates with a high degree of discrimination (0.997 D value).The same multilocus genotype was shared by isolates recovered from some patients and from the hand of theirs correspondent healthcare worker. For another patient, the same multilocus genotype of C. metapsilosis was detected in blood and skin confirming that candidemia usually arises as an endogenous infection following prior colonization.

Conclusions

These STR markers are a valuable tool for the differentiation of C. parapsilosis complex strains, to support epidemiological investigations especially studies of strain relatedness and pathways of transmission.

     

Properties of Allyl Alcohol-Resistant Mutants of Clostridium butyricum Grown on Glycerol

Mutants of Clostridium butyricum E5 exhibiting resistance to allyl alcohol which produced the same quantities of 1,3-propanediol as the wild-type strain but more acetate than butyrate were isolated. The acetate-butyrate formation plays a major function in the regulation of the internal redox balance. Allyl alcohol resistance can be attributed not to the loss of 1,3-propanediol dehydrogenase but to a shift in the reductive properties of the enzyme. The data support the view that cellular regulation is modified to avoid intracellular accumulation of 3-hydroxypropionaldehyde.     

Insights into the Na+ Storage Mechanism of Phosphorus‐Functionalized Hard Carbon as Ultrahigh Capacity Anodes

Hard carbon as a typical anode material for sodium ion batteries has received much attention in terms of its low cost and renewability. Herein, phosphorus‐functionalized hard carbon with a specific “honeycomb briquette” shaped morphology is synthesized via electrospinning technology. When applied as an anode material for Na⁺ storage, it exhibits an impressively high reversible capacity of 393.4 mA h g^?1 with the capacity retention up to 98.2% after 100 cycles. According to first‐principle calculation, the ultrahigh capacity of the as‐prepared anode is ascribed to the enhancement of Na‐absorption through formation of P?O and P? C bonds in graphitic layers when doped with phosphorus. Moreover, the increase of electron density around the Fermi level is found to be mainly caused by O atoms instead of P atoms.     

Physiological relevance of plant 2‐Cys peroxiredoxin overoxidation level and oligomerization status

Peroxiredoxins are ubiquitous thioredoxin‐dependent peroxidases presumed to display, upon environmental constraints, a chaperone function resulting from a redox‐dependent conformational switch. In this work, using biochemical and genetic approaches, we aimed to unravel the factors regulating the redox status and the conformation of the plastidial 2‐Cys peroxiredoxin (2‐Cys PRX) in plants. In Arabidopsis, we show that in optimal growth conditions, the overoxidation level mainly depends on the availability of thioredoxin‐related electron donors, but not on sulfiredoxin, the enzyme reducing the 2‐Cys PRX overoxidized form. We also observed that upon various physiological temperature, osmotic and light stress conditions, the overoxidation level and oligomerization status of 2‐Cys PRX can moderately vary depending on the constraint type. Further, no major change was noticed regarding protein conformation in water‐stressed Arabidopsis, barley and potato plants, whereas species‐dependent up‐ and down‐variations in overoxidation were observed. In contrast, both 2‐Cys PRX overoxidation and oligomerization were strongly induced during a severe oxidative stress generated by methyl viologen. From these data, revealing that the oligomerization status of plant 2‐Cys PRX does not exhibit important variation and is not tightly linked to the protein redox status upon physiologically relevant environmental constraints, the possible in planta functions of 2‐Cys PRX are discussed.     

Zinner’s syndrome: report of two cases and review of the literature

Background

Congenital malformations of the seminal vesicle are uncommon, and most of them are cystic malformations. If an insult occurs between the 4th and the 13 h gestational week, the embryogenesis of the kidney, ureter, seminal vesicle, and vas deferens could be altered. Cysts of the seminal vesicle may appear with a mass effect, dysuria, epididymitis, or obstruction of the gastrointestinal and genitourinary tracts. Approximately two thirds of them are associated with ipsilateral renal agenesis, because both the ureteral buds and seminal vesicles originate from the mesonephric (Wolffian) duct. They were first described by Zinner in 1914, and 200 cases of seminal vesicle cysts associated with ipsilateral renal agenesis have been reported in the literature. Most patients with this anomaly are asymptomatic until the third or fourth decade of life. Some cases have nonspecific symptoms such as prostatism, urinary urgency, dysuria, painful ejaculation, and perineal discomfort. Transrectal ultrasonography provides good visualization of the pelvic structures and allows guidance for aspiration of the cysts.

Case presentation

We present two cases of seminal vesicle cyst. The first patient had dysuria, increased frequency of urination, and haematuria. He was operated and benefited from a removal of the cyst with right ureterectomy and left ureteral reimplantation. The second patient had disorder of the digestive transit and he benefited from a laparoscopic removal of the cyst.

Conclusions

Seminal vesicle cysts combined with ipsilateral renal agenesis are rare urological anomalies. Usual symptoms that are caused by the seminal vesicle cysts are bladder irritation and obstruction as well as pain in the perineum and scrotum. Epididymitis is frequently found. Treatment consists to removing the seminal vesicle cyst.

     

Detection of African horse sickness virus in Culicoides imicola pools using RT‐qPCR

African horse sickness (AHS) is an infectious, non‐contagious arthropod‐borne disease of equids, caused by the African horse sickness virus (AHSV), an orbivirus of the Reoviridae family. It is endemic in sub‐Saharan Africa and thought to be the most lethal viral disease of horses. This study focused on detection of AHSV in Culicoides imicola (Diptera: Ceratopogonidae) pools by the application of a RT‐qPCR. Midges were fed on AHSV‐infected blood. A single blood‐engorged female was allocated to pools of unfed nulliparous female midges. Pool sizes varied from 1 to 200. RNA was extracted and prepared for RT‐qPCR. The virus was successfully detected and the optimal pool size for the limit of detection of the virus was determined at a range between 1 to 25. Results from this investigation highlight the need for a standardized protocol for AHSV investigation in Culicoides midges especially for comparison among different studies and for the determination of infection rate.