An Optical Fiber-Based Nanomotion Sensor for Rapid Antibiotic and Antifungal Susceptibility Tests.

The emergence of antibiotic and antifungal resistant microorganisms represents nowadays a major public health issue that might push humanity into a post-antibiotic/antifungal era. One of the approaches to avoid such a catastrophe is to advance rapid antibiotic and antifungal susceptibility tests. In this study, we present a compact, optical fiber-based nanomotion sensor to achieve this goal by monitoring the dynamic nanoscale oscillation of a cantilever related to microorganism viability. High detection sensitivity was achieved that was attributed to the flexible two-photon polymerized cantilever with a spring constant of 0.3 N/m. This nanomotion device showed an excellent performance in the susceptibility tests of Escherichia coli and Candida albicans with a fast response in a time frame of minutes. As a proof-of-concept, with the simplicity of use and the potential of parallelization, our innovative sensor is anticipated to be an interesting candidate for future rapid antibiotic and antifungal susceptibility tests and other biomedical applications.

Deciphering the human antibody response against Burkholderia pseudomallei during melioidosis using a comprehensive immunoproteome approach.

Introduction: The environmental bacterium Burkholderia pseudomallei causes the often fatal and massively underreported infectious disease melioidosis. Antigens inducing protective immunity in experimental models have recently been identified and serodiagnostic tools have been improved. However, further elucidation of the antigenic repertoire of B. pseudomallei during human infection for diagnostic and vaccine purposes is required. The adaptation of B. pseudomallei to very different habitats is reflected by a huge genome and a selective transcriptional response to a variety of conditions. We, therefore, hypothesized that exposure of B. pseudomallei to culture conditions mimicking habitats encountered in the human host might unravel novel antigens that are recognized by melioidosis patients. Methods and results: In this study, B. pseudomallei was exposed to various stress and growth conditions, including anaerobiosis, acid stress, oxidative stress, iron starvation and osmotic stress. Immunogenic proteins were identified by probing two-dimensional Western blots of B. pseudomallei intracellular and extracellular protein extracts with sera from melioidosis patients and controls and subsequent MALDI-TOF MS. Among B. pseudomallei specific immunogenic signals, 90 % (55/61) of extracellular immunogenic proteins were identified by acid, osmotic or oxidative stress. A total of 84 % (44/52) of intracellular antigens originated from the stationary growth phase, acidic, oxidative and anaerobic conditions. The majority of the extracellular and intracellular protein antigens were identified in only one of the various stress conditions. Sixty-three immunoreactive proteins and an additional 38 candidates from a literature screening were heterologously expressed and subjected to dot blot analysis using melioidosis sera and controls. Our experiments confirmed melioidosis-specific signals in 58 of our immunoproteome candidates. These include 15 antigens with average signal ratios (melioidosis:controls) greater than 10 and another 26 with average ratios greater than 5, including new promising serodiagnostic candidates with a very high signal-to-noise ratio. Conclusion: Our study shows that a comprehensive B. pseudomallei immunoproteomics approach, using conditions which are likely to be encountered during infection, can identify novel antibody targets previously unrecognized in human melioidosis.

Microbial Sampling Using Interdental Brushes and Paper Points around Teeth and Implants: A Pilot Study for Comparison.

Inflammatory periodontal and peri-implant diseases follow dysbiotic shifts in a susceptible host. A well-established tool for microbial sample collection is the use of paper points. The purpose of this pilot study was to evaluate the use of interdental brushes compared to paper points. Biofilm samples were collected with paper points and later interdental brushes from ten patients. Five patients were represented with a community periodontal index of treatment needs (CPITN) of 0-2 around the teeth and an implant with PPD ≤ 5 mm and no radiological bone loss. The remaining five patients had a CPITN ≥ 3 and one implant with peri-implantitis. Microbial samples were analyzed with quantitative polymerase chain reaction (qPCR) and next-generation sequencing (NGS). The results showed higher amounts of DNA in samples taken by interdental brushes but also higher Ct values. Both methods detected Filifactor alocis, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, and Treponema denticola in the majority of samples, while Aggregatibacter actinomycetemcomitans was rarely found. A microbial dysbiosis index showed comparable or higher values in sites with no periodontitis/peri-implantitis with interdental brushes. The results of this pilot study indicate that interdental brushes might be a valid technique for microbial sampling and particularly advantageous in the early detection of dysbiotic shifts around teeth and implants. Larger studies with more participants are needed to validate the proposed microbial sampling method with interdental brushes.

Compilation of different data sets of the Late Neolithic wetland site of Pestenacker and of the adjacent valley depositions.

This document contains data sets of the valley depositions of the Loosbach valley and data of the Late Neolithic wetland site of Pestenacker. It consists of raw data and graphical figures of direct push-based electrical conductivity and colour logs and driving core recoveries as well as hand drilling recoveries presented by Köhler et al. [1]. We reviewed unpublished archaeological profiles to determine the incision levels of former stream phases at Pestenacker site. Here, we provide the new, reusable and accessible data set. The data sets and figures of the valley depositions can be used for further analyses, including statistical ones, to improve the methods of the direct-push sensing and to compare it with the sedimentological features recovered from driving core and hand drillings. In addition, the data set is useful for further issues in Pestenacker as well as in the whole central Europe. Especially in the circum-Alpine region, as a comparison with other pile dwellings or stilt houses built from the Neolithic to the Bronce Age.

Evaluating ACGME-accredited addiction psychiatry fellowship online content: A critical analysis of addiction psychiatry fellowship program websites in the US.

Introduction: There is an extreme shortage of addiction psychiatrists and a lack of representation of addiction psychiatry (ADP) fellows from racial/ethnic minoritized backgrounds. ADP fellowship websites are integral in engaging potential applicants. It is therefore critical to understand the quality of engagement that trainees are having with ADP fellowship websites. The aim of this study was to investigate the accessibility and content of ADP fellowship program websites in the U.S. Methods: A list of ADP Fellowship programs was obtained from the Accreditation Council for Graduate Medical Education. A critical textual analysis of 42 unique factors within four categories (accessibility, recruitment, education, and health equity) was performed for each ADP fellowship website. Results: Of 51 ADP fellowships, 47 (92.2%) had websites. Information about social media accounts was largely missing from ADP fellowship websites. For recruitment, program description (95.7%) and program director name (76.6%) were most readily available, while interview day (0.00%) and vacation details (10.6%) were least available. For education, a list of rotations (55.3%) and didactics/lectures (40.4%) were most readily available, while post fellowship placement (6.4%), call schedule (4.3%), and responsibility progression (2.1%) were least available. The most prevalent health equity factors were gender-inclusive language (100%) and an absence of stigmatizing addiction language (100%). The least listed were statements of commitment to health equity (0.0%), antiracism training (2.1%), and harm-reduction strategies (4.3%). Conclusions: There are considerable gaps in the amount and types of information provided by ADP fellowship websites. Many existing websites are poorly interfacing with potential leaders in the field. The development of ADP fellowship websites could serve as a low-cost recruitment tool to engage potential addiction specialists. Our findings underscore the need for ADP fellowships to optimize their websites to engage bourgeoning leaders in addiction and optimize access to more comprehensive information.

Perceptions on navigating ACGME-accredited addiction psychiatry fellowship program websites: A thematic analysis across a race- and gender-diverse pool of potential applicants.

Background: There is an alarming shortage of addiction psychiatrists in the United States. To promote interest in addiction psychiatry (ADP), it is essential to maximize resources available through ADP fellowship websites. The aim of this study was to investigate the perceived adequacy and accessibility of content on ADP fellowship websites and discover what further information is considered important among trainees interested in becoming addiction specialists. Methods: Three virtual focus groups were conducted between January and February 2021 among medical students and residents in diverse geographic regions. Participants were asked about the availability of information on ADP fellowship program websites and other material they would like to see available. Focus groups were recorded, with data transcribed and coded using NVivo 11 and Dedoose. A coding scheme was deductively developed based on the core research questions. Results: The majority of participants (N = 27) identified areas of dissatisfaction with the content currently available on ADP websites. The sample was highly representative of racial and ethnic minoritized trainees (n = 12) and genderqueer/non-binary participants (n = 3). Three major themes were identified and durable across all focus groups: lack of emphasis on diversity/health equity, lack of portrayal of everyday life and activities of fellows, and inadequate representation of curricula. Overwhelmingly, participants identified a dedication to health equity (for example, working with minoritized populations) as a key deciding factor in whether to apply to a particular ADP fellowship. Conclusions: ADP fellowship websites are perceived to have considerable variability in the amount and quality of information. Many do not appear to provide the full spectrum of content desired by diverse potential applicants, such as information regarding current fellows and community-centered initiatives. This is concerning, as it suggests ADP fellowships may be interfacing poorly with burgeoning leaders, especially those from race and gender minoritized backgrounds, neglecting potential opportunities to develop future addiction specialists.

Identifying proteins bound to native mitotic ESC chromosomes reveals chromatin repressors are important for compaction.

Epigenetic information is transmitted from mother to daughter cells through mitosis. Here, to identify factors that might play a role in conveying epigenetic memory through cell division, we report on the isolation of unfixed, native chromosomes from metaphase-arrested cells using flow cytometry and perform LC-MS/MS to identify chromosome-bound proteins. A quantitative proteomic comparison between metaphase-arrested cell lysates and chromosome-sorted samples reveals a cohort of proteins that were significantly enriched on mitotic ESC chromosomes. These include pluripotency-associated transcription factors, repressive chromatin-modifiers such as PRC2 and DNA methyl-transferases, and proteins governing chromosome architecture. Deletion of PRC2, Dnmt1/3a/3b or Mecp2 in ESCs leads to an increase in the size of individual mitotic chromosomes, consistent with de-condensation. Similar results were obtained by the experimental cleavage of cohesin. Thus, we identify chromosome-bound factors in pluripotent stem cells during mitosis and reveal that PRC2, DNA methylation and Mecp2 are required to maintain chromosome compaction.

Correction to: Nucleus replacement could get a new chance with annulus closure.

Unfortunately, Fig. 7 and last paragraph of the result section have been incorrectly published. The complete corrected Fig. 7 and last paragraph of the results part (IDP measurements) have been as follows.

Nucleus replacement could get a new chance with annulus closure.

PURPOSE: Disc herniations are usually treated by decompression of the spinal nerves via a partial nucleotomy. As a consequence of reduced disc height (DH), reduced intradiscal pressure (IDP) and increased range of motion (ROM), accelerated degeneration may occur. Nucleus replacement implants are intended to restore those values, but are associated with the risk of extrusion. METHODS: In six fresh frozen lumbar spinal segments (L2-3/L3-4/L4-5/L5-S1, age median 64.5 years (57-72), Pfirrmann grade 2-3), a prolapse was provoked through a box defect (6 × 10 mm) in the annulus. The herniated nucleus material was removed and replaced by a novel collagen-based nucleus implant. An annulus closure device sealed the defect. ROM, neutral zone (NZ) and IDP were measured in the (1) intact and (2) defect state, (3) postoperatively and (4) after cyclic loading (n = 100,000 cycles) applying pure moments (± 7.5 Nm) in flexion-extension, lateral bending and axial rotation. Additionally, the change in DH was determined. Extrusion of implants or nucleus material was evaluated macroscopically. RESULTS: In all specimens, a prolapse could be provoked which decreased DH. Subsequent nucleotomy changed ROM/NZ and IDP considerably. Initial values could be restored by the implantation. Macroscopically, none of the implants nor nucleus material did migrate after cyclic loading. CONCLUSIONS: In this study, a prolapse followed by a nucleotomy resulted in a biomechanical destabilisation. Implantation of the nucleus replacement combined with an annulus closure restored the intact condition without showing signs of extrusion nor migration after cyclic loading. Hence, nucleus replacements could have a new chance in combination with annulus closure devices.

A perspective view on the nanomotion detection of living organisms and its features.

The insurgence of newly arising, rapidly developing health threats, such as drug-resistant bacteria and cancers, is one of the most urgent public-health issues of modern times. This menace calls for the development of sensitive and reliable diagnostic tools to monitor the response of single cells to chemical or pharmaceutical stimuli. Recently, it has been demonstrated that all living organisms oscillate at a nanometric scale and that these oscillations stop as soon as the organisms die. These nanometric scale oscillations can be detected by depositing living cells onto a micro-fabricated cantilever and by monitoring its displacements with an atomic force microscope-based electronics. Such devices, named nanomotion sensors, have been employed to determine the resistance profiles of life-threatening bacteria within minutes, to evaluate, among others, the effect of chemicals on yeast, neurons, and cancer cells. The data obtained so far demonstrate the advantages of nanomotion sensing devices in rapidly characterizing microorganism susceptibility to pharmaceutical agents. Here, we review the key aspects of this technique, presenting its major applications. and detailing its working protocols.

Evaluation of disinfectant efficacy against multidrug-resistant bacteria: A comprehensive analysis of different methods.

BACKGROUND: Multidrug-resistant gram-negative bacteria (MDR-GNB) constitute a threat to health care worldwide. Disinfectants are used to prevent and control the spread of MDR-GNB in a hospital setting but their efficacy might be impaired by bacterial mechanisms that may act on both antimicrobials and disinfectants. Determination of minimum inhibitory concentrations is mainly used to determine bacterial susceptibility against disinfectants, but practical tests on surfaces might be more suitable to predict in-use conditions. Our objective was to compare and evaluate 4 different methods widely used to assess surface disinfectant efficacy. METHODS: The efficacy of benzalkonium chloride (BAC), peracetic acid (PAA), and ethanol (ETH) against multidrug-resistant Acinetobacter, Pseudomonas, and Klebsiella strains was assessed by minimum inhibitory concentration determinations, quantitative suspension tests, qualitative suspension tests, and carrier tests. Test results were compared to ascertain the most appropriate method. RESULTS: ETH, PAA, and BAC were highly effective against MDR-GNB, but we observed marked differences in efficacious concentrations (up to 100-fold) as a function of the test method applied. Minimum inhibitory concentration determination was not reliable for evaluating susceptibility or resistance to BAC. CONCLUSIONS: Surface tests should be used to determine bacterial susceptibility against disinfectants. Moreover, suitable guidelines are needed that allow for the standardization and comparison of bactericidal values obtained by different investigators.

Low Diagnostic Yield of Routine Cerebrospinal Fluid Analysis in Juvenile Stroke.

Background: The diagnostic value of cerebrospinal fluid (CSF) analysis in juvenile stroke, i.e., stroke in young adult patients, is not well studied. We sought to determine the therapeutic impact of routine CSF-analysis in young adults with acute ischemic stroke or transient ischemic attack (TIA). Methods: We abstracted data from patients with acute cerebral ischemia aged 18-45 years who were consecutively admitted to our stroke center between 01/2008 and 12/2015. We routinely performed CSF-analysis in patients with hitherto unknown stroke etiology after complete diagnostic work up. We assessed the frequency and underlying causes of abnormal CSF-findings and their impact on secondary stroke prevention therapy. Results: Among 379 patients (median [IQR:IQR3-IQR1] age 39 [10:43-33] years, 48% female) with acute ischemic stroke (n = 306) or TIA (n = 73), CSF analysis was performed in 201 patients (53%). Of these, 25 patients (12.4 %) had CSF pleocytosis (leucocyte cell count ≥ 5 Mpt/L), that was rated as non-specific (e.g., traumatic lumbar puncture, reactive pleocytosis) in 22 patients. Only 3 patients (1.5% of all patients who underwent CSF-analysis) with CSF-pleocytosis had specific CSF-findings that were related to stroke etiology and affected secondary stroke prevention therapy. Imaging findings had already suggested cerebral vasculitis in two of these patients. Conclusions: The diagnostic yield of routine CSF-analysis in juvenile stroke was remarkably low in our study. Our data suggest that CSF-analysis should only be performed if further findings raise the suspicion of cerebral vasculitis.

An Additive Manufacturing Technique for the Facile and Rapid Fabrication of Hydrogel-based Micromachines with Magnetically Responsive Components.

Polyethylene glycol (PEG)-based hydrogels are biocompatible hydrogels that have been approved for use in humans by the FDA. Typical PEG-based hydrogels have simple monolithic architectures and often function as scaffolding materials for tissue engineering applications. More sophisticated structures typically take a long time to fabricate and do not contain moving components. This protocol describes a photolithography method that allows for facile and rapid microfabrication of PEG structures and devices. This strategy involves an in-house developed fabrication stage that allows for the rapid fabrication of 3D structures by building upwards in a layer-by-layer fashion. Independent moving components can also be aligned and assembled onto support structures to form integrated devices. These independent components are doped with superparamagnetic iron oxide nanoparticles that are sensitive to magnetic actuation. In this manner, the fabricated devices can be actuated using external magnets to yield movement of the components within. Hence, this technique allows for the fabrication of sophisticated MEMS-like devices (micromachines) that are composed entirely out of a biocompatible hydrogel, able to function without an onboard power source, and respond to a contact-less method of actuation. This manuscript describes the fabrication of both the fabrication set-up as well as the step-by-step method for the microfabrication of these hydrogels-based MEMS-like devices.

Allele-specific analysis of cell fusion-mediated pluripotent reprograming reveals distinct and predictive susceptibilities of human X-linked genes to reactivation.

BACKGROUND: Inactivation of one X chromosome is established early in female mammalian development and can be reversed in vivo and in vitro when pluripotency factors are re-expressed. The extent of reactivation along the inactive X chromosome (Xi) and the determinants of locus susceptibility are, however, poorly understood. Here we use cell fusion-mediated pluripotent reprograming to study human Xi reactivation and allele-specific single nucleotide polymorphisms (SNPs) to identify reactivated loci. RESULTS: We show that a subset of human Xi genes is rapidly reactivated upon re-expression of the pluripotency network. These genes lie within the most evolutionary recent segments of the human X chromosome that are depleted of LINE1 and enriched for SINE elements, predicted to impair XIST spreading. Interestingly, this cadre of genes displays stochastic Xi expression in human fibroblasts ahead of reprograming. This stochastic variability is evident between clones, by RNA-sequencing, and at the single-cell level, by RNA-FISH, and is not attributable to differences in repressive histone H3K9me3 or H3K27me3 levels. Treatment with the DNA demethylating agent 5-deoxy-azacytidine does not increase Xi expression ahead of reprograming, but instead reveals a second cadre of genes that only become susceptible to reactivation upon induction of pluripotency. CONCLUSIONS: Collectively, these data not only underscore the multiple pathways that contribute to maintaining silencing along the human Xi chromosome but also suggest that transcriptional stochasticity among human cells could be useful for predicting and engineering epigenetic strategies to achieve locus-specific or domain-specific human Xi gene reactivation.

Additive manufacturing of hydrogel-based materials for next-generation implantable medical devices.

Implantable microdevices often have static components rather than moving parts, and exhibit limited biocompatibility. This paper demonstrates a fast manufacturing method which can produce features in biocompatible materials down to tens of microns in scale, with intricate and composite patterns in each layer. By exploiting unique mechanical properties of hydrogels, we developed a "locking mechanism" for precise actuation and movement of freely moving parts, which can provide functions such as valves, manifolds, rotors, pumps, and delivery of payloads. Hydrogel components could be tuned within a wide range of mechanical and diffusive properties, and can be controlled after implantation without a sustained power supply. In a mouse model of osteosarcoma, triggering of release of doxorubicin from the device over ten days showed high treatment efficacy and low toxicity, at one-tenth of a standard systemic chemotherapy dose. Overall, this platform, called "iMEMS", enables development of biocompatible implantable microdevices with a wide range of intricate moving components that can be wirelessly controlled on demand, in a manner that solves issues of device powering and biocompatibility.

Human isotype-dependent inhibitory antibody responses against Mycobacterium tuberculosis.

Accumulating evidence from experimental animal models suggests that antibodies play a protective role against tuberculosis (TB). However, little is known about the antibodies generated upon Mycobacterium tuberculosis (MTB) exposure in humans. Here, we performed a molecular and functional characterization of the human B-cell response to MTB by generating recombinant monoclonal antibodies from single isolated B cells of untreated adult patients with acute pulmonary TB and from MTB-exposed healthcare workers. The data suggest that the acute plasmablast response to MTB originates from reactivated memory B cells and indicates a mucosal origin. Through functional analyses, we identified MTB inhibitory antibodies against mycobacterial antigens including virulence factors that play important roles in host cell infection. The inhibitory activity of anti-MTB antibodies was directly linked to their isotype. Monoclonal as well as purified serum IgA antibodies showed MTB blocking activity independently of Fc alpha receptor expression, whereas IgG antibodies promoted the host cell infection. Together, the data provide molecular insights into the human antibody response to MTB and may thereby facilitate the design of protective vaccination strategies.

Rapid and Sensitive Multiplex Detection of Burkholderia pseudomallei-Specific Antibodies in Melioidosis Patients Based on a Protein Microarray Approach.

BACKGROUND: The environmental bacterium Burkholderia pseudomallei causes the infectious disease melioidosis with a high case-fatality rate in tropical and subtropical regions. Direct pathogen detection can be difficult, and therefore an indirect serological test which might aid early diagnosis is desirable. However, current tests for antibodies against B. pseudomallei, including the reference indirect haemagglutination assay (IHA), lack sensitivity, specificity and standardization. Consequently, serological tests currently do not play a role in the diagnosis of melioidosis in endemic areas. Recently, a number of promising diagnostic antigens have been identified, but a standardized, easy-to-perform clinical laboratory test for sensitive multiplex detection of antibodies against B. pseudomallei is still lacking. METHODS AND PRINCIPAL FINDINGS: In this study, we developed and validated a protein microarray which can be used in a standard 96-well format. Our array contains 20 recombinant and purified B. pseudomallei proteins, previously identified as serodiagnostic candidates in melioidosis. In total, we analyzed 196 sera and plasmas from melioidosis patients from northeast Thailand and 210 negative controls from melioidosis-endemic and non-endemic regions. Our protein array clearly discriminated between sera from melioidosis patients and controls with a specificity of 97%. Importantly, the array showed a higher sensitivity than did the IHA in melioidosis patients upon admission (cut-off IHA titer ≥1:160: IHA 57.3%, protein array: 86.7%; p = 0.0001). Testing of sera from single patients at 0, 12 and 52 weeks post-admission revealed that protein antigens induce either a short- or long-term antibody response. CONCLUSIONS: Our protein array provides a standardized, rapid, easy-to-perform test for the detection of B. pseudomallei-specific antibody patterns. Thus, this system has the potential to improve the serodiagnosis of melioidosis in clinical settings. Moreover, our high-throughput assay might be useful for the detection of anti-B. pseudomallei antibodies in epidemiological studies. Further studies are needed to elucidate the clinical and diagnostic significance of the different antibody kinetics observed during melioidosis.

Experimental fetal endoscopic tracheal occlusion in rhesus and cynomolgus monkeys: nonhuman primate models.

OBJECTIVE: The monkey model is the best model to investigate some physiological response to the fetal transitory tracheal occlusion but it has never been described in Macaca monkeys. The aim of this study was to evaluate the feasibility of fetal endoscopic tracheal occlusion (FETO) in a non-human primate model. METHODS: Pregnant rhesus monkeys and cynomolgus were tested as a potential experimental model for FETO in the third trimester of pregnancy, by performing fetal tracheoscopies with and without tracheal occlusion. RESULTS: A total of 22 pregnancies were followed in 16 monkeys and underwent fetal surgery. Percutaneous endoscopic access to the uterine cavity was possible in 20 cases (91%). Of these 20 pregnant monkeys, fetal tracheoscopy could be achieved in 15 cases (75%). In rhesus monkeys, the time between the onset of endoscopy and tracheal penetration decreases as operator experience increases. Neither maternal morbidity nor mortality was related to surgery. Two fetal losses were possibly due to the procedure. CONCLUSION: FETO is feasible in the non-human primate, which closely reflects procedures in humans. The non-human primate model for FETO, specially the rhesus monkeys, may be useful for future studies concerning the mechanisms related to the lung growth after transitory fetal tracheal occlusion.

Reactive oxygen species and excitation-contraction coupling in the context of cardiac pathology.

Reactive oxygen species (ROS) are highly reactive oxygen-derived chemical compounds that are by-products of aerobic cellular metabolism as well as crucial second messengers in numerous signaling pathways. In excitation-contraction-coupling (ECC), which links electrical signaling and coordinated cardiac contraction, ROS have a severe impact on several key ion handling proteins such as ion channels and transporters, but also on regulating proteins such as protein kinases (e.g. CaMKII, PKA or PKC), thereby pivotally influencing the delicate balance of this finely tuned system. While essential as second messengers, ROS may be deleterious when excessively produced due to a disturbed balance in Na(+) and Ca(2+) handling, resulting in Na(+) and Ca(2+) overload, SR Ca(2+) loss and contractile dysfunction. This may, in the end, result in systolic and diastolic dysfunction and arrhythmias. This review aims to provide an overview of the single targets of ROS in ECC and to outline the role of ROS in major cardiac pathologies, such as heart failure and arrhythmogenesis. This article is part of a Special Issue entitled "Redox Signalling in the Cardiovascular System"

Puf operon sequences and inferred structures of light-harvesting complexes of three closely related Chromatiaceae exhibiting different absorption characteristics.

Whole cells of the purple sulfur bacterium strain 970 exhibit an unusual absorption peak at 963 nm. Its closest relatives, Thiorhodovibrio (Trv.) winogradskyi DSM6702(T) and strain 06511 display a bacteriochlorophyll (BChl) a absorption peak at 867 nm that is characteristic for most light-harvesting complexes 1 (LHC1) of proteobacteria. The puf operons encoding the LHC1 and reaction center proteins were amplified, cloned, and sequenced, and for the Trv. winogradskyi, strains show the common pufBALMC gene arrangement, whereas strain 970 contains a second pufBA copy downstream of pufC. Only pufB(1)A(1) is transcribed, and the corresponding mRNA fragment had an increased stability. Alignments of the deduced protein sequences showed that the LHC1 polypeptides are closely related to those of Thermochromatium (Tch.) tepidum. A deletion between αHis(0) and αTrp(+11), thought to be responsible for the redshifted Q(y) absorption in Tch. tepidum, was also detected in strain 970 and Trv. winogradskyi, whereas αLys(+12) is replaced by histidine only in strain 970. Based on our structural modeling, the side chain of this αHis is predicted to be in close proximity to the BChl a, suggesting that it exerts a modulating effect on the spectral properties of the highly unusual LHC1 complex of strain 970.