Cutibacterium acnes
is an abundant skin commensal with several proposed mutualistic functions. A protein with strong antioxidant activity was recently identified from the
C
.
acnes
secretome. This protein, termed RoxP, facilitated aerobic bacterial growth
in vitro
and
ex vivo
. As reducing events naturally occurred outside of the bacterial cell, it was further hypothesized that RoxP could also serve to modulate redox status of human skin. The biological function of RoxP was here assessed
in vitro
and
in vivo
, through oxidatively stressed cell cultures and through protein quantification from skin affected by oxidative disease (actinic keratosis and basal cell carcinoma), respectively. 16S rDNA amplicon deep sequencing and single locus sequence typing was used to correlate bacterial prevalence to cutaneous RoxP abundances. We show that RoxP positively influence the viability of monocytes and keratinocytes exposed to oxidative stress, and that a congruent concentration decline of RoxP can be observed in skin affected by oxidative disease. Basal cell carcinoma was moreover associated with microbial dysbiosis, characterized by reduced
C
.
acnes
prevalence.
C
.
acnes
’s secretion of RoxP, an exogenous but naturally occurring antioxidant on human skin, is likely to positively influence the human host. Results furthermore attest to its prospective usability as a biopharmaceutical.
Bdellovibrio and like organisms (BALOs) are obligate predatory bacteria that selectively prey on a broad range of Gram-negative bacteria, including multidrug-resistant human pathogens. Due to their unique lifestyle, they have been long recognized as a potential therapeutic and biocontrol agent. Research on BALOs has rapidly grown over the recent decade, resulting in many publications concerning molecular details of bacterial predation as well as applications thereof in medicine and biotechnology. This review summarizes the current knowledge on biotechnological potential of obligate predatory bacteria and their secreted enzymes.
In
this study, a surface plasmon resonance (SPR) biosensor
was developed for the detection and quantification of a secreted bacterial
factor (RoxP) from skin. A molecular imprinting method was used for
the preparation of sensor chips and five different monomer-cross-linker
compositions were evaluated for sensitivity, selectivity, affinity,
and kinetic measurements. The most promising molecularly imprinted
polymer (MIP) was characterized by using scanning electron microscopy,
atomic force microscopy, and cyclic voltammetry. Limit of detection
(LOD) value was calculated as 0.23 nM with an affinity constant of
3.3 × 10–9 M for the promising MIP. Besides
being highly sensitive, the developed system was also very selective
for the template protein RoxP, proven by the calculated selectivity
coefficients. Finally, absolute concentrations of RoxP in several
skin swabs were analyzed by using the developed MIP-SPR biosensor
and compared to a competitive ELISA. Consequently, the developed system
offers a very efficient tool for the detection and quantification
of RoxP as an early indicator for some oxidative skin diseases especially
when they are present in low-abundance levels (e.g., skin samples).
Stapylococcus aureus is a common infectious agent in e.g. sepsis, associated with both high mortality rates and severe long-term effects. The cytolytic protein α-hemolysin has repeatedly been shown to enhance the virulence of S. aureus. Combined with an unhindered spread of multi drug-resistant strains, this has triggered research into novel anti virulence (i.e. anti α-hemolysin) drugs. Their functionality will depend on our ability to identify infections that might be alleviated by such. We therefore saw a need for detection methods that could identify individuals suffering from S. aureus infections where α-hemolysin was a major determinant. Molecular imprinted polymers were subsequently prepared on gold coated sensor chips. Used in combination with a surface plasmon resonance biosensor, α-hemolysin could therethrough be quantified from septic blood samples (n = 9), without pre-culturing of the infectious agent. The biosensor recognized α-hemolysin with high affinity (KD = 2.75 x 10-7 M) and demonstrated a statistically significant difference (p < 0.0001) between the α-hemolysin response and potential sample contaminants. The detection scheme proved equally good, or better, when compared to antibody-based detection methods. This novel detection scheme constitutes a more rapid, economical, and user-friendly alternative to many methods currently in use. Heightening both reproducibility and sensitivity, molecular imprinting in combination with surface plasmon resonance (SPR)-technology could be a versatile new tool in clinical- and research-settings alike.
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