Heterologous expression of NoxA confers aerotolerance in Clostridium sporogenes.

Clostridium is a genus of gram-positive obligate anaerobic bacteria. Some species of Clostridium, including Clostridium sporogenes, may be of use in bacteria-mediated cancer therapy. Spores of Clostridium are inert in healthy normoxic tissue but germinate when in the hypoxic regions of solid tumors, causing tumor regression. However, such treatments fail to completely eradicate tumors partly because of higher oxygen levels at the tumor's outer rim. In this study, we demonstrate that a degree of aerotolerance can be introduced to C. sporogenes by transfer of the noxA gene from Clostridium aminovalericum. NoxA is a water-forming NADH oxidase enzyme, and so has no detrimental effect on cell viability. In addition to its potential in cancer treatment, the noxA-expressing strain described here could be used to alleviate challenges related to oxygen sensitivity of C. sporogenes in biomanufacturing.

Hemodialysis-Associated Immune Dysregulation in SARS-CoV-2-Infected End-Stage Renal Disease Patients.

Patients on hemodialysis show dysregulated immunity, basal hyperinflammation and a marked vulnerability to COVID-19. We evaluated the immune profile in COVID-19 hemodialysis patients and the changes associated with clinical deterioration after the hemodialysis session. Recruited patients included eight hemodialysis subjects with active, PCR-confirmed SARS-CoV-2 infection, five uninfected hemodialysis patients and five healthy controls. In SARS-CoV-2-infected hemodialysis patients TNF-α, IL-6 and IL-8 were particularly increased. Lymphopenia was mostly due to reduction in CD4+ T, B and central memory CD8+ T cells. There was a predominance of classical and intermediate monocytes with reduced HLA-DR expression and enhanced production of pro-inflammatory molecules. Immune parameters were analysed pre- and post-hemodialysis in three patients with COVID-19 symptoms worsening after the hemodialysis session. There was a higher than 2.5-fold increase in GM-CSF, IFN-γ, IL-1ß, IL-2, IL-6, IL-17A and IL-21 in serum, and augmentation of monocytes-derived TNF-α, IL-1ß and IL-8 and CXCL10 (p < 0.05). In conclusion, COVID-19 in hemodialysis patients associates with alteration of lymphocyte subsets, increasing of pro-inflammatory cytokines and monocyte activation. The observed worsening during the hemodialysis session in some patients was accompanied by augmentation of particular inflammatory cytokines, which might suggest biomarkers and therapeutic targets to prevent or mitigate the hemodialysis-related deterioration during SARS-CoV-2 infection.

Haemolytic uraemic syndrome associated with pancreatitis: report of four cases and review of the literature.

Background: The incidence of acute kidney injury (AKI) in patients with acute pancreatitis ranges from 15% to 40% and is associated with poor prognosis. Haemolytic uraemic syndrome (HUS) in the setting of acute pancreatitis is an uncommon association with fewer than 30 cases reported in the literature. Methods: A retrospective review of the clinical records at our institution between January 1981 and December 2019 was carried out to identify patients with acute pancreatitis and HUS. Additionally, a literature review was conducted on this topic. The aims of the study were to describe the clinical course and outcomes of patients affected by this condition. Results: Four cases of HUS following an acute pancreatitis were identified. The mean (±SD) age of the study group was 30 ± 6 years, all of which were males. Excessive alcohol consumption was the main cause of acute pancreatitis in all four patients. HUS with progressive AKI developed in a median interval of 2 days from the onset of pancreatitis (range 1-3 days). All patients required kidney replacement therapy during the course of follow-up. A kidney biopsy was performed in two patients, showing typical thrombotic microangiopathic features. One case was treated with eculizumab, whereas the rest were treated with supportive care and/or plasma exchange. A normalization of haematological parameters and complete recovery of kidney function were observed in all patients at last follow-up, although this improvement was significantly faster in the patient treated with eculizumab. Conclusions: HUS may infrequently develop in patients with acute pancreatitis. An early identification of this complication is mandatory, and complement blockade with eculizumab may be associated with a faster kidney function recovery.

Erratum: Haemolytic uraemic syndrome associated with pancreatitis: report of four cases and review of the literature.

[This corrects the article DOI: 10.1093/ckj/sfaa245.][This corrects the article DOI: 10.1093/ckj/sfaa245.].

Pericarditis secundaria a infección por COVID-19 en un paciente trasplantado renal / Pericarditis secondary to COVID-19 infection in a kidney transplant recipient

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Complete Genome Sequence of Enterococcus thailandicus Strain a523 Isolated from Urban Raw Sewage.

We report here the first complete circularly closed genome sequence of Enterococcus thailandicus strain a523 isolated from raw urban sewage. This genome contains 2,646,250 bp with a G+C content of 36.8%, 2,499 genes, 2,370 protein-coding sequences, 6 rRNA operons, 65 tRNAs, and 6 clustered regularly interspaced short palindromic repeat arrays.

False Negative Results in Clostridium difficile Testing.

BACKGROUND: Accurate diagnosis of Clostridium difficile infection (CDI) is paramount for patient management. The wrong diagnosis places patients at risk, delays treatment, and/ or contributes to transmission of infection in the healthcare setting. Although amplification of the toxin B gene by polymerase chain reaction (PCR) is a sensitive method for detecting toxigenic C. difficile, false negative results still occur and could impact the diagnosis and treatment of this infection. METHODS: This study investigated 48 patients that tested negative for toxigenic C. difficile via GeneXpert C. difficile epi test, while simultaneously testing positive for toxigenic C. difficile via stool culture. Fifty discrepant samples were collected over a 15-month period and all C. difficile isolates were characterized by ribotype. Patient charts were reviewed to assess whether discrepant results impacted the treatment course or clinical outcome of affected patients. RESULTS: Fifty samples of a total of 2308 samples tested in an acute healthcare facility over a 15-month period had negative PCR and positive stool culture for toxigenic C. difficile. C. difficile isolated from the discrepant samples resulted in diverse ribotyping patterns suggesting they were derived from different strains. The samples belonged to patients who were distributed evenly between age groups and wards in the hospital. In the majority of cases, the false negative C. difficile test results did not seem to impact the clinical outcome in these patients. CONCLUSIONS: The PCR limit of detection may impact the results of molecular methods for C. difficile detection. Both clinical and analytical sensitivity of C. difficile tests should be considered when deciding which diagnostic assay to use, and clinical correlates should be examined carefully before excluding CDI as a cause of disease.

Impact of polymerase chain reaction testing on Clostridium difficile infection rates in an acute health care facility.

Two rapid methods of Clostridium difficile infection (CDI) diagnosis were compared between June 2012 and March 2013: a GeneXpert (Cepheid, Sunnyvale, Calif) polymerase chain reaction (PCR) test and an enzyme immunoassay (EIA). The influence of these methods on the detection of hospital-acquired CDI and identification of CDI outbreaks was evaluated. We tested 1,592 stool samples for C difficile. The GeneXpert PCR test identified 211 positive samples (68 determined to be hospital-acquired infection), whereas EIA identified 105 positive samples (36 determined to be hospital-acquired infection). The GeneXpert PCR method in contrast to the EIA method increased the detection rates of nosocomial CDI cases and contributed to the declaration of CDI outbreaks.

Non-Markovian near-infrared Q branch of HCl diluted in liquid Ar.

By using a non-Markovian spectral theory based in the Kubo cumulant expansion technique, we have qualitatively studied the infrared Q branch observed in the fundamental absorption band of HCl diluted in liquid Ar. The statistical parameters of the anisotropic interaction present in this spectral theory were calculated by means of molecular dynamics techniques, and found that the values of the anisotropic correlation times are significantly greater (by a factor of two) than those previously obtained by fitting procedures or microscopic cell models. This fact is decisive for the observation in the theoretical spectral band of a central Q resonance which is absent in the abundant previous researches carried out with the usual theories based in Kubo cumulant expansion techniques. Although the theory used in this work only allows a qualitative study of the Q branch, we can employ it to study the unknown characteristics of the Q resonance which are difficult to obtain with the quantum simulation techniques recently developed. For example, in this study we have found that the Q branch is basically a non-Markovian (or memory) effect produced by the spectral line interferences, where the PR interferential profile basically determines the Q branch spectral shape. Furthermore, we have found that the Q resonance is principally generated by the first rotational states of the first two vibrational levels, those more affected by the action of the dissolvent.

Clospore: a liquid medium for producing high titers of semi-purified spores of Clostridium difficile.

Clostridium difficile continues to cause infections in healthcare and other settings. Its spores survive well indoors and require sporicidal chemicals for infection control. However, proper testing of disinfectants is impeded due to difficulties in obtaining viable spores of high enough quality and titers to meet current regulations for sporicidal claims. A new liquid medium (Clospore) has been developed, based on a systematic review of the compositions of 20 other available media. C. difficile spores grown in the new medium and treated with a mixture of lysozyme and trypsin yielded final suspensions with > 10(9) CFU/mL of viable spores, with a purity of > 91% as tested by spore-staining and phase-contrast microscopy. The spores showed a biological decay rate of about 0.1 log10/month when dried on metal disks and stored indoors (air temperature 23 +/- 2 degrees C; relative humidity 52.76 +/- 15.08%). Heating the purified spore suspensions to 70 degrees C for 10 min to inactivate any vegetative cells showed no spore activation or inactivation. The spores could be stored for at least 14 months either refrigerated (4 degrees C) or frozen (-20 or -80 degrees C) in 50% (v/v) ethanol with virtually no loss in viability. The resistance of the enzyme-treated spores to three levels of sodium hypochlorite (1000, 3000, and 5000 ppm), using a standardized quantitative carrier test, was almost identical to that of the spores concentrated by centrifugation alone. The described procedure has been successfully applied to four standard (ATCC) and six clinical strains of C. difficile.

Diffuse reflectance spectroscopy characterization of hemoglobin and intralipid solutions: in vitro measurements with continuous variation of absorption and scattering.

Absorption and scattering processes in biological tissues are studied through reflectance spectroscopy in tissue-like phantoms. For this aim, an experimental setup is designed to independently control both processes in hemoglobin and intralipid solutions. From the analysis of the obtained spectra, a simple empirical power law equation is found that relates absorbance with scattering and absorption coefficients. This relationship includes three wavelength independent parameters, which can be determined geometry from in vitro measurements for each particular optical optode. The dependence of the optical path length on the absorption and scattering coefficients is also analyzed, and estimations of this parameter for physiological conditions are presented. This study is useful to better understand the scattering phenomena in biological tissue, and to obtain absolute concentration of absorber particles when a homogeneous medium can be assumed.

Activity of selected oxidizing microbicides against the spores of Clostridium difficile: relevance to environmental control.

BACKGROUND: Clostridium difficile is an increasingly common nosocomial pathogen, and its spores are resistant to common environmental surface disinfectants. Many high-level disinfectants (eg, aldehydes) are unsuitable for environmental decontamination because they need several hours of contact to be sporicidal. This study tested the potential of selected oxidative microbicides to inactivate C. difficile spores on hard surfaces in relatively short contact times at room temperature. METHODS: The spores of a clinical isolate of C. difficile were tested using disks (1 cm diameter) of brushed stainless steel in a quantitative carrier test. The spores of C. sporogenes and Bacillus subtilis, common surrogates for evaluating sporicides, were included for comparison. The clostridia were grown separately in Columbia broth (CB), and B. subtilis was grown in a 1:10 dilution of CB. Each disk received 10 microL test spores with an added soil load, and the inoculum was dried. One disk each was placed in a glass vial and overlaid with 50 microL test formulation; controls received an equivalent volume of normal saline with 0.1% Tween 80. At the end of the contact time the microbicide was neutralized, the inoculum recovered from the disks by vortexing, the eluates were membrane filtered, and the filters placed on plates of recovery medium. The colony-forming units (CFU) on the plates were recorded after 5 days of incubation. The performance criterion was > or = 6 log(10) (> or = 99.9999%) reduction in the viability titer of the spores. The microbicides tested were domestic bleach with free-chlorine (FC) levels of 1000, 3000, and 5000 mg/L; an accelerated hydrogen peroxide (AHP)-based product with 70,000 mg/L H2O2 (Virox STF); chlorine dioxide (600 mg/L FC); and acidified domestic bleach (5000 mg/L FC). RESULTS: Acidified bleach and the highest concentration of regular bleach tested could inactivate all the spores in < or = 10 minutes; Virox STF could do the same in < or = 13 minutes. Regular bleach with 3000 mg/L FC required up to 20 minutes to reduce the viability of the all the spores tested to undetectable levels; chlorine dioxide and the lowest concentration of regular bleach tested needed approximately 30 minutes for the same level of activity. CONCLUSIONS: Acidified bleach, Virox STF, and regular bleach (3000-5000 mg/L FC) could inactivate C. difficile spores on hard environmental surfaces in approximately 10 to 15 minutes under ambient conditions. All of these products are strong oxidizers and should be handled with care for protection of staff, but acidified and regular bleach with high levels of FC also release chlorine gas, which can be hazardous if inhaled by staff or patients.