Humoral responses to Cytomegalovirus glycoprotein B vaccine with MF59 adjuvant

Solid organ transplant (SOT) patients are at risk of end-organ diseases such as pneumonitis, hepatitis or enteritis caused by HCMV. HCMV infection can occur via primary infection of a seronegative recipient or upon reinfection or reactivation in a seropositive transplant recipient. Seronegative recipients have the greatest risk of viraemia and disease, showing that pre-existing natural immunity provides substantial protection. This, in turn, underpins vaccination as a viable strategy to control HCMV in the transplant setting. To test this, a clinical trial with a vaccine based on HCMV glycoprotein B (gB) antigen plus MF59 adjuvant was performed in SOT awaiting transplantation. The study showed that antibody titres against the gB antigen were significantly increased in both seropositive and seronegative recipients of the vaccine in comparison to the patients who received placebo, and importantly, higher titres correlated directly with reduced viraemia post-transplant. The aim of my thesis was to identify the component of the specific humoral response responsible for this effect. In comparison with placebo recipients, I could find no evidence for the protection being due to induction of antibodies that mediate neutralisation, antibody dependent cellular cytotoxicity, or prevent cell to cell spread of virus in culture. In contrast, analysis of antigenic domains of gB bound by the antibodies revealed that vaccination of seropositive individuals enhanced antibody responses against antigenic domain 2 and that these correlated with reduced viraemia post-transplant. Antibodies against three other antigenic domains were induced by the vaccine, but did not correlate with protection. These results suggest that antigenic domain 2 should be an important component of future HCMV vaccines to boost pre-existing immune responses that protect from HCMV infection. The protection afforded to seronegatives remains unidentified, but could be explained if another antigenic domain on gB remains to be discovered

Characterization of two new CTX-M-25-group extended-spectrum β-lactamase variants identified in Escherichia coli isolates from Israel.

OBJECTIVES: We characterized two new CTX-M-type extended-spectrum β-lactamase (ESBL) variants in Escherichia coli isolates from stool samples of two elderly patients admitted at the Tel Aviv Sourasky Medical Center, Israel. Both patients underwent treatment with cephalosporins prior to isolation of the E. coli strains. METHODS: ESBLs were detected by the double-disk synergy test and PCR-sequencing of β-lactamase genes. The bla(CTX-M) genes were cloned into the pCR-BluntII-TOPO vector in E. coli TOP10. The role of amino-acid substitutions V77A and D240G was analyzed by site-directed mutagenesis of the bla(CTX-M-94) and bla(CTX-M-100) genes and comparative characterization of the resulting E. coli recombinants. MICs of β-lactams were determined by Etest. Plasmid profiling, mating experiments, replicon typing and sequencing of bla(CTX-M) flanking regions were performed to identify the genetic background of the new CTX-M variants. RESULTS: The novel CTX-M β-lactamases, CTX-M-94 and -100, belonged to the CTX-M-25-group. Both variants differed from CTX-M-25 by the substitution V77A, and from CTX-M-39 by D240G. CTX-M-94 differed from all CTX-M-25-group enzymes by the substitution F119L. Glycine-240 was associated with reduced susceptibility to ceftazidime and leucine-119 with increased resistance to ceftriaxone. bla(CTX-M-94) and bla(CTX-M-100) were located within ISEcp1 transposition units inserted into ∼93 kb non-conjugative IncFI and ∼130 kb conjugative IncA/C plasmids, respectively. The plasmids carried also different class 1 integrons. CONCLUSIONS: This is the first report on CTX-M-94 and -100 ESBLs, novel members of the CTX-M-25-group

A global multinational survey of cefotaxime-resistant coliforms in urban wastewater treatment plants

The World Health Organization Global Action Plan recommends integrated surveillance programs as crucial strategies for monitoring antibiotic resistance. Although several national surveillance programs are in place for clinical and veterinary settings, no such schemes exist for monitoring antibiotic-resistant bacteria in the environment. In this transnational study, we developed, validated, and tested a low-cost surveillance and easy to implement approach to evaluate antibiotic resistance in wastewater treatment plants (WWTPs) by targeting cefotaxime-resistant (CTX-R) coliforms as indicators. The rationale for this approach was: i) coliform quantification methods are internationally accepted as indicators of fecal contamination in recreational waters and are therefore routinely applied in analytical labs; ii) CTX-R coliforms are clinically relevant, associated with extended-spectrum ?-lactamases (ESBLs), and are rare in pristine environments. We analyzed 57 WWTPs in 22 countries across Europe, Asia, Africa, Australia, and North America. CTX-R coliforms were ubiquitous in raw sewage and their relative abundance varied significantly (< 0.1% to 38.3%), being positively correlated (p < 0.001) with regional atmospheric temperatures. Although most WWTPs removed large proportions of CTX-R coliforms, loads over 103 colony-forming units per mL were occasionally observed in final effluents. We demonstrate that CTX-R coliform monitoring is a feasible and affordable approach to assess wastewater antibiotic resistance status

Antioxidant, Anti-Inflammatory, and Postulated Cytotoxic Activity of Phenolic and Anthocyanin-Rich Fractions from Polana Raspberry (Rubus idaeus L.) Fruit and Juice—In Vitro Study

In this study, the antioxidative and anti-inflammatory potential of crude extracts (CE), anthocyanin-rich fractions (ARF), and phenolic fractions (PF) from raspberry (R) and raspberry juice (J) were evaluated. The antioxidant properties were evaluated with three complementary assays: DPPH radical scavenging activity, chelating Fe(II) power, and ferric reducing power. The highest antioxidant activity was determined for the crude extract from raspberry pulp (RCE) in the case of all methods used. The anti-inflammatory activity was demonstrated by inhibitory effect on lipoxygenase (LOX) and cyclooxygenase-2 (COX-2) activity in vitro. The highest efficiency in inhibiting the activity of both enzymes was exhibited by RCE, 0.79 and 0.59 mg FW/mL, respectively. In turn, JARF had the lowest ability to inhibit LOX (EC50 = 4.5 mg FW/mL) and JPF caused the lowest COX-2 inhibition (1.75 mg FW/mL). Additionally, we have performed a pilot study of in vitro cytotoxic activity using two human leukemia cell lines: J45 and HL60. All examined extracts inhibited the viability of J45 cells more effectively than HL60. The highest cytotoxic effect was observed in the J45.01 cell line after exposure to RCE (EC50 = 0.0375 mg FW/mL)

Etnolog na rynku pracy

Prezentowana publikacja powstała w oparciu o materiał, jaki powstał podczas zajęć warsztatowych pt. „Etnolog na rynku pracy” prowadzonych w roku akademickim 2011/2012 przez doktor Annę Weronikę Brzezińską w Instytucie Etnologii i Antropologii Kulturowej Uniwersytetu im. Adama Mickiewicza w Poznaniu. Do jej napisania zaprosiliśmy uczestników warsztatów – studentów I oraz II roku studiów II stopnia (studia magisterskie) na kierunku etnologia . Założeniem zajęć było rozpoznanie rynku pracy pod kątem studiów etnologicznych – jego specyfiki, potrzeb pracodawców, najbardziej przydatnych umiejętności oraz wiedzy, niezbędnej w późniejszym rozwoju kariery zawodowej. Dzięki przeprowadzonym przez nas badaniom udało nam się cel ten nie tylko spełnić, ale i przekroczyć. Niniejszą książkę potraktować można nie tylko jako zbiór informacji o stanie rynku pracy w kontekście antropologii kulturowej, ale również jako poradnik dla studentów, chcących świadomie zaplanować swoją ścieżkę zawodową. Z drugiej strony może ona pełnić rolę informatora dla potencjalnych pracodawców, mających wątpliwości co do kwalifikacji zawodowych humanistów. Czytelnik dowie się o słabościach i silnych stronach absolwentów etnologii, a przede wszystkim o zmianach jakie zaszły w procesie dydaktycznym

Two Different Extended-Spectrum β-Lactamases (ESBLs) in One of the First ESBL-Producing Salmonella Isolates in Poland

Two extended-spectrum β-lactamase (ESBL)-producing salmonella isolates, Salmonella enterica serovar Enteritidis and Salmonella enterica serovar Typhimurium, were analyzed. Both isolates produced the CTX-M-3 ESBL; however, their bla(CTX-M-3) genes were located on different plasmids. The serovar Typhimurium isolate also expressed another ESBL, SHV-2a, and probably the two ESBL genes had been acquired independently by the strain

Impact of the Nucleic Acid Extraction Method and the RT-qPCR Assay on SARS-CoV-2 Detection in Low-Viral Samples

COVID-19 was initially reported in China at the end of 2019 and soon thereafter, in March 2020, the WHO declared it a pandemic. Until October 2021, over 240 million COVID-19 cases were recorded, with 4.9 mln deaths. In order to stop the spread of this disease, it is crucial to monitor and detect any infected person. The etiologic agent of COVID-19 is a novel coronavirus called SARS-CoV-2. The gold standard for the detection of the virus is the RT-qPCR method. This study evaluated two RNA extraction methods and four commercial RT-qPCR assays routinely used in diagnostic laboratories for detecting SARS-CoV-2 in human specimens from the upper respiratory tract. We analyzed a panel of 70 clinical samples with varying RNA loads. Our study demonstrated the significant impact of the diagnostic methods selected by the laboratory on the SARS-CoV-2 detection in clinical specimens with low viral loads