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Preliminary microbiological diagnosis usually relies on microscopic examination and, due to the routine culture and bacteriological examination, lasts up to 11 days. Hence, many deep learning methods based on microscopic images were recently introduced to replace the time-consuming bacteriological examination. They shorten the diagnosis by 1-2 days but still require iterative culture to obtain monoculture samples. In this work, we present a feasibility study for further shortening the diagnosis time by analyzing polyculture images. It is possible with multi-MIL, a novel multi-label classification method based on multiple instance learning. To evaluate our approach, we introduce a dataset containing microscopic images for all combinations of four considered bacteria species. We obtain ROC AUC above 0.9, proving the feasibility of the method and opening the path for future experiments with a larger number of species.

Industrial production of biogas offers a way to manage distillery leachate. The waste is usually subjected to anaerobic digestion for producing biogas. However, the effluent from anaerobic processes has high chemical oxygen demand (COD) and is harmful to the environment. An effective method of lowering COD is ozonation. Effluent from biogas plants after ozonation has the potential for use in breeding grounds for plants of the Lemnaceae family. Thus, they can provide a valuable additional source of biomass for the production of bioethanol. Lemna minor L. and Spirodela polyrhiza cultures were grown in media with the addition of 2.5% PFE, which had been treated by ozonation for between 6 and 50 min. Using ozonated effluent was an effective cultivation technique in all variants. The analyzed parameters were plant growth, chlorophyll index, fresh plant weight and photosynthetic traits (net photosynthesis, stomatal conductance, transpiration and concentration of intercellular CO2). The best growth of Lemna minor L. was observed in the media with PFE treated for 12 min. Similar effects were obtained for S. polyrhiza, with ozone treatment for 12 and 25 min. The results show the potential of using ozone-treated post-fermentation leachate as a supplement in culture media.

Background:In recent years, a dynamic increase in the incidence of oropharyngeal squamous cell carcinoma (OPSCC) has been observed, especially in highly developed countries. As in the case of anal and genital cancers, a significant proportion of OPSCC (about 60%) is due to sexually acquired HPV infection, and the increase in OPSCC is attributed to changes in sexual behavior in Western countries. EBV and CMV viruses are also characterized by carcinogenesis, but the literature data on their participation in the development of neoplasms of the middle throat are still very limited.Objective:The aim of this article is to review the latest scientific reports on the impact of infections with latent viruses (HPV, EBV, CMV) on the oncogenesis process in the middle pharynx and to summarize the prophylactic options using the available multivalent vaccines against HPV infections.Conclusions:HPV-positive oropharyngeal squamous cell carcinoma of the middle pharynx is a significant clinical problem due to the increase in the number of cases, especially in young people, who lack classic risk factors, such as exposure to tobacco and alcohol. HPV-dependent neoplasms are characterized by a better prognosis and less invasive oncological therapy (radiotherapy and chemotherapy). Of central importance in the prevention of HPV-positive OPSCC in both women and men is prophylaxis with multivalent vaccination. Due to the lack of sufficient data on the participation of EBV and CMV viruses in the development of OPSCC, as well as the lack of prophylaxis, intensive research in this area is necessary.

The lag phase is a temporary non-replicative period observed when a microbial population is introduced to a new nutrient-rich environment. Its duration can have a pronounced effect on population fitness, and it is often measured in laboratory conditions. However, calculating the lag phase length may be challenging and method and parameters dependent. Moreover, the details of these methods and parameters used throughout experimental studies are often under-reported. Here we discuss the most frequently used methods in experimental and theoretical studies, and we point out some inconsistencies between them. Using experimental and simulated data we study the performance of these methods depending on the frequency of population size measurements, and parameters determining the growth curve shape, such as growth rate. It turns out that the sensitivity to each of these parameters depends on the lag calculation methods. For example, lag duration calculation by parameter fitting to a logistic model is very robust to low frequency of measurements, but it may be highly biased for growth curves with low growth rate. On the contrary, the method based on finding the point where growth acceleration is the highest, is robust to low growth rate, but highly sensitive to low frequency of measurements and the level of noise in the data. Based on our results, we propose a decision tree to choose a method most suited to one’s data. Finally, we developed a web tool where the lag duration can be calculated based on the user-specified growth curve data, and for various explicitly specified methods, parameters, and data pre-processing techniques.GRAPHICAL ABSTRACT

Introduction: The coronavirus disease 2019 (COVID-19) is one of the greatest clinical challenges of the last decades. Clinical factors associated with severity of the disease remain unclear. The aim of the study was to characterize Polish patients hospitalized due to COVID-19 and to evaluate potential prognostic factors of severe course of the disease. Material and methods: An observational study was conducted from March to July 2020 in the Pulmonology and Allergology Department of the University Hospital in Kraków, Poland. Consecutive patients with confirmed SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) infection were enrolled, and data about past medical history, signs and symptoms, laboratory results, imaging studies results, in-hospital management and outcomes was prospectively gathered. Results: The study sample comprised 100 patients at the mean age of 59.2 (SD 16.1) years among whom 63 (63.0%) were male. Among them 10 (10.0%) died, 47 (47%) presented respiratory failure, 15 (15.0%) were transferred to the intensive care unit, 17 (17.0%) developed acute kidney injury, 7 (7.0%) had sepsis and 10 (10.0%) were diagnosed with pulmonary embolism. Multivariable analysis revealed age (OR 1.1; 95% CI 1.01–1.15), body mass index (BMI; OR 1.24; 95% CI 1.01–1.53), modified early warning score (MEWS; OR 3.95; 95% CI 1.48–12), the highest d-dimer value (OR 1.73; 95% CI 1.03–2.9) and lactate dehydrogenase (LDH; OR 1.16; 95% CI 1.03–1.3) to be associated with severe course of COVID-19. Conclusion: This observational study showed that almost half of hospitalized patients with COVID-19 developed respiratory failure in the course of the disease. Increasing age, BMI, MEWS, d-dimer value and LDH concentration were associated with the severity of COVID-19.

Hydrogels derived from TEMPO-oxidised cellulose nanofibrils (TOCNs) are not robust and inherently water unstable if the TOCNs are not crosslinked or coated with a water-swellable polymer. Furthermore, the manufacturing of self-standing TOCN films is still a challenge due to the small TOCN diameter and viscosifying effect. Here, we report the TEMPO-mediated oxidation of bacterial cellulose (BC) nanopaper as a route to produce robust and water stable TOCN hydrogel without the need for additional additives or crosslinking steps, as well as self-standing TOCN films without the need for vacuum filtration or slow-drying of TOCN suspension. Pristine BC pellicle was first press-dried into a dried and well-consolidated BC nanopaper, followed by TEMPO-oxidation at various NaClO concentrations. The oxidation reaction introduced carboxylate moieties onto exposed BC nanofibrils within the nanopaper network structure. This then led to the swelling of the nanopaper into a hydrogel. A swelling ratio of up to 100 times the original thickness of BC nanopaper was observed upon TEMPO-oxidation. The water retention value of the TEMPO-oxidised BC hydrogels was also found to increase with increasing carboxylate content. These TEMPO-oxidised BC hydrogels were found to be robust and water-stable, even under prolonged (>1 month) magnetic stirring in water. We further showed that high grammage self-standing TOCN films (100 g m-2) can be fabricated as simple as press-drying a water stable TEMPO-oxidised BC hydrogels without the need of vacuum-assisted filtration or slow-drying, which is typically the rate-limiting step in the manufacturing of self-standing TOCN films.

Most cells spend the majority of their life in the non-proliferating, quiescent state. Transition to this state is crucial for microorganisms to survive long starvation periods and restart divisions afterwards. Experimental evolution allowed us to identify several mutation in genes that are presumably important for such transition in yeast cells. Most of these candidate genes belong to the SPS amino acid sensing pathway or to the SIR complex. We assembled these mutations on the ancestral strain background. Analysis of the quiescent/non-quiescent cell ratio of the starved yeast populations confirmed the crucial role of SSY1, the primary receptor component of the SPS sensor, in transition to the Q state. The evolved SSY1 mutations increased yeast sensitivity to amino acid presence in the environment. This resulted in decreased quiescent cell fraction and a 5.14% increase of the total amino acid content in the starved populations. We discuss external amino acid sensing via the SPS pathway as one of the mechanisms influencing transition to quiescence.

Background Accidental awareness during general anaesthesia (AAGA) is a complex and rare outcome to investigate in surgical patient populations, particularly obstetric patients. We report the protocol of the Direct Reporting of Awareness in Maternity patients (DREAMY) study, illustrating how the research was designed to address practical and methodological challenges for investigating AAGA in an obstetric cohort. Methods This is the trial protocol of a prospective, multicentre cohort study of patients undergoing obstetric surgery under general anaesthesia. Accidental awareness during general anaesthesia will be detected using three repetitions of standardised direct questioning over 30 days, with responses indicating memories during general anaesthesia verified using structured interviews. Reports will be adjudicated, then classified, in accordance with pre-defined and pre-validated structures, including the Michigan Awareness Classification tool. Quantitative data will be collected on general anaesthesia conduct for all participants. This descriptive study is being conducted in England and aims to recruit a minimum of 2015 patients. Results The DREAMY study was prospectively registered (ClinicalTrials.gov Identifier: NCT03100396) and ethical approval granted. Participant recruitment began in May 2017 and one year follow up concluded in August 2019. Publication of the results is anticipated in 2020. Conclusions The DREAMY study will provide data on incidence, experience and implications of AAGA for obstetric patients, using a robust methodology that will reliably detect and translate subjective AAGA reports into objective outcomes. In addition, the study is expected to improve vigilance for AAGA in participating hospitals and encourage adoption of recommendations for support of patients experiencing AAGA.

Background Breast cancer (BC) risk assessment models are statistical estimates based on patient characteristics. We developed a gene expression assay to assess BC risk using benign breast biopsy tissue. Methods A NanoString-based malignancy risk (MR) gene signature was validated for formalin-fixed paraffin-embedded (FFPE) tissue. It was applied to FFPE benign and BC specimens obtained from women who underwent breast biopsy, some of whom developed BC during follow-up to evaluate diagnostic capability of the MR signature. BC risk was calculated with MR score, Gail risk score, and both tests combined. Logistic regression and receiver operating characteristic curves were used to evaluate these 3 models. Results NanoString MR demonstrated concordance between fresh frozen and FFPE malignant samples (r = 0.99). Within the validation set, 563 women with benign breast biopsies from 2007 to 2011 were identified and followed for at least 5 y; 50 women developed BC (affected) within 5 y from biopsy. Three groups were compared: benign tissue from unaffected and affected patients and malignant tissue from affected patients. Kruskal–Wallis test suggested difference between the groups (P = 0.09) with trend in higher predicted MR score for benign tissue from affected patients before development of BC. Neither the MR signature nor Gail risk score were statistically different between affected and unaffected patients; combining both tests demonstrated best predictive value (AUC = 0.71). Conclusions FFPE gene expression assays can be used to develop a predictive test for BC. Further investigation of the combined MR signature and Gail Model is required. Our assay was limited by scant cellularity of archived breast tissue.

Group B streptococcus (GBS) is one of the uropathogens that causes urinary tract infections (UTIs). The aims of this article were molecular characterization, an analysis of antimicrobial susceptibility profiles, adherence to bladder endothelial cells, and the detection of immunoreactive proteins of 94 clinical strains of GBS isolated from adult Polish patients with UTI. Antibiotic susceptibilities were determined by disk diffusion. Serotyping and Alp family genes detection were studied using multiplex PCR. Genetic profiles were determined by pulsed-field gel electrophoresis. The adherence ability of the studied strains was estimated by incubation on human bladder microvascular endothelial cell line. Immunoreactive proteins were studied by immunoblotting. Antibiotic susceptibility investigation revealed that 22% of GBS strains were resistant to erythromycin, whereas 18% demonstrated resistance to clindamycin. cMLSB was present in 76% of the resistant strains, M phenotype was detected in 14%, whereas iMLSB was present for 10%. The most common serotype was serotype III (31%), followed by serotype V (27%), and serotype Ia (17%). The genes that dominated among other Alp genes were: epsilon (29%), alp2 (27%), and rib (23%). The most common co-occurring serotypes and Alp genes were: Ia and epsilon, III and rib, III and alp2, V and alp2, and V and alp3 (p < 0.001). The PFGE method showed high clonality for serotype V and cMLSB (p < 001). The PFGE method showed high clonality for serotype V. Furthermore, this serotype was significantly associated with the cMLSB phenotype (p < 0.001). The most common immunoreactive proteins demonstrated masses of 50 kDa and 45–47 kDa. Although examined GBS isolates showed high genetic diversity, immunoreactive proteins were common for most of the studied GBS isolates, which may indicate their conservation, and allows to consider them as potential immunodiagnostic markers. Although the examined GBS isolates showed high genetic diversity, immunoreactive proteins were shared by most of the studied GBS isolates. It may indicate their conservation, thus allowing to consider them as potential immunodiagnostic markers.

This article applies four-wave mixing (FWM) spectroscopy, a third-order nonlinear optical spectroscopic technique which is not intrinsically surface- or interface-sensitive, to study silicone oil thin films, supplemented by second-order nonlinear-optical sum frequency generation (SFG) vibrational spectroscopy. Although studies of thin organic films using coherent antistokes Raman spectroscopy (CARS), a special case of FWM, have been reported previously, in this study we demonstrate the feasibility of using a more general FWM process which involves three independent excitation laser beams to investigate silicone oil thin films. The results show that the FWM method has the potential to detect and provide molecular-level information on ultrathin silicone oil layers, down to a film thickness of 1 nm. This developed FWM methodology is widely applicable and can be utilized to study important issues in the biopharmaceutical field, e.g., to examine the distribution of silicone oil on syringe glass surfaces with subnanometer sensitivity. It can also be used to study the potentially slow reactions between silicone oil and glass surfaces as proposed in the literature but without direct molecular-level information.

OBJECTIVE To assess the incidence of maternal group B Streptococcus (GBS) infection in England. DESIGN Population surveillance augmented through data linkage. SETTING England. POPULATION All pregnant women accessing the National Health Service (NHS) in England. METHODS Invasive GBS (iGBS) infections during pregnancy or within 6 weeks of childbirth were identified by linking Public Health England (PHE) national microbiology surveillance data for 2014 to NHS hospital admission records. Capsular serotypes of GBS were determined by reference laboratory typing of clinical isolates from women aged 15-44 years. Post-caesarean section surgical site infection (SSI) caused by GBS was identified in 21 hospitals participating in PHE SSI surveillance (2009-2015). MAIN OUTCOME MEASURES iGBS rate per 1000 maternities; risk of GBS SSI per 1000 caesarean sections. RESULTS Of 1601 patients diagnosed with iGBS infections in England in 2014, 185 (12%) were identified as maternal infections, a rate of 0.29 (95% CI 0.25-0.33) per 1000 maternities and representing 83% of all iGBS cases in women aged 18-44 years. Seven (3.8%) were associated with miscarriage. Fetal outcome identified excess rates of stillbirth (3.4 versus 0.5%) and extreme prematurity (

To persist in variable environments populations of microorganisms have to survive periods of starvation and be able to restart cell division in nutrient-rich conditions. Typically, starvation signals initiate a transition to a quiescent state in a fraction of individual cells, while the rest of the cells remain non-quiescent. It is widely believed that, while quiescent cells (Q) help the population to survive long starvation, the non-quiescent cells (NQ) are a side effect of imperfect transition. We analysed regrowth of starved monocultures of Q and NQ cells compared to mixed, heterogeneous cultures in simple and complex starvation environments. Our experiments, as well as mathematical modelling, demonstrate that Q monocultures benefit from better survival during long starvation, and from a shorter lag phase after resupply of rich medium. However, when the starvation period is very short, the NQ monocultures outperform Q and mixed cultures, due to their short lag phase. In addition, only NQ monocultures benefit from complex starvation environments, where nutrient recycling is possible. Our study suggests that phenotypic heterogeneity in starved populations could be a form of bet hedging, which is adaptive when environmental determinants, such as the length of the starvation period, the length of the regrowth phase, and the complexity of the starvation environment vary over time.ImportanceNon-genetic cell heterogeneity is present in glucose starved yeast populations in the form of quiescent (Q) and nonquiescent (NQ) phenotypes. There is evidence that Q cells help the population to survive long starvation. However, the role of the NQ cell type is not known, and it has been speculated that the NQ phenotype is just a side effect of imperfect transition to the Q phenotype. Here we show that, in contrast, there are ecological scenarios in which NQ cells perform better than monocultures of Q cells or naturally occuring mixed populations containing both Q and NQ. NQ cells benefit when the starvation period is very short and environmental conditions allow nutrient recycling during starvation. Our experimental and mathematical modeling results suggest a novel hypothesis: the presence of both Q and NQ phenotypes within starved yeast populations may reflect a form of bet hedging, where different phenotypes provide fitness advantages depending on environmental conditions.

Contains fulltext : 240254.pdf (Publisher’s version ) (Open Access) 22 november 2021

Currently, most cotton textile waste is sent to landfill. However, due to the use of synthetic additives and the chemical treatment of cotton fibers, cotton textile waste is difficult to biodegrade. Cotton textile waste can also be subjected to material recycling, or to incineration/gasification to produce energy. Here, we present the optimization of acid hydrolysis of cotton yarn fibers for glucose efficiency. The cotton yarn hydrolysates showed great potential for replacing simple sugar solutions in fermentation media. The highest glucose concentration was obtained in the hydrolysates of cotton yarn hydrolyzed in a 2% solution of sulfuric acid or phosphoric acid at 140–160 °C for 2 h. After 2 h of hydrolysis at 140 °C with 2% H3PO4, the concentration of glucose in the cotton yarn hydrolysate (13.19 g/L) increased fivefold compared with cotton yarn treated under the same conditions with H2SO4 (2.65 g/L). The structural modifications in the solid residues after acid hydrolysis were analyzed using a scanning electron microscope with energy dispersive spectroscopy (SEM-EDS), attenuated total reflectance Fourier-transform infrared spectroscopy (FTIR-ATR), and Raman spectroscopy. The SEM images, IR spectra, and Raman spectra revealed that the most significant changes in the morphology of the fibers occurred when the process was carried out at high temperatures (≥140 °C). Better growth of the yeast strains Saccharomyces cerevisiae Ethanol Red and Saccharomyces cerevisiae Tokay ŁOCK0204 was observed in the medium containing phosphoric acid hydrolysate. The maximum methane yield of 278 dm3/kgVS and the maximum hydrogen yield of 42 dm/kgVS were reported for cotton yarn waste after pretreatment with H3PO4. This might have been linked to the beneficial effect of phosphorus, which is a key nutrient for anaerobic digestion. The proposed hydrolysis method does not generate fermentation inhibitors.

Microorganisms are famous for adapting quickly to new environments. However, most evidence for rapid microbial adaptation comes from laboratory experiments or domesticated environments, and it is unclear how rates of adaptation scale from human‐influenced environments to the great diversity of wild microorganisms. We examined potential monthly‐scale selective pressures in the model forest yeast Saccharomyces paradoxus. Contrary to expectations of seasonal adaptation, the S. paradoxus population was stable over four seasons in the face of abiotic and biotic environmental changes. While the S. paradoxus population was diverse, including 41 unique genotypes among 192 sampled isolates, there was no correlation between S. paradoxus genotypes and seasonal environments. Consistent with observations from other S. paradoxus populations, the forest population was highly clonal and inbred. This lack of recombination, paired with population stability, implies that selection is not acting on the forest S. paradoxus population on a seasonal timescale. Saccharomyces paradoxus may instead have evolved generalism or phenotypic plasticity with regard to seasonal environmental changes long ago. Similarly, while the forest population included diversity among phenotypes related to intraspecific interference competition, there was no evidence for active coevolution among these phenotypes. At least ten percent of the forest S. paradoxus individuals produced “killer toxins,” which kill sensitive Saccharomyces cells, but the presence of a toxin‐producing isolate did not predict resistance to the toxin among nearby isolates. How forest yeasts acclimate to changing environments remains an open question, and future studies should investigate the physiological responses that allow microbial cells to cope with environmental fluctuations in their native habitats., We have sampled populations of the wild yeast species Saccharomyces paradoxus in a natural forest to study spatial and temporal variation. The distribution of genetic and phenotypic diversity was analyzed using microsatellite markers and in vitro assays. We show that yeast populations are characterized by stability, high dispersal potential, clonality, and inbreeding, while still including genetic and phenotypic diversity.

Highly collaborative scientists are often called on to extend their expertise to different types of projects and to expand the scope and scale of projects well beyond their previous experience. For a large-scale project involving "big data" to be successful, several different aspects of the research plan need to be developed and tested, which include but are not limited to the experimental design, sample collection, sample preparation, metadata recording, technical capability, data acquisition, approaches for data analysis, methods for integration of different data types, recruitment of additional expertise as needed to guide the project, and strategies for clear communication throughout the project. To capture this process, we describe an example project in proteogenomics that built on our collective expertise and experience. Key steps included definition of hypotheses, identification of an appropriate clinical cohort, pilot projects to assess feasibility, refinement of experimental designs, and extensive discussions involving the research team throughout the process. The goal of this chapter is to provide the reader with a set of guidelines to support development of other large-scale multiomics projects.

The evolutionary transition from single-celled to multicellular growth is a classic and intriguing problem in biology. Saccharomyces cerevisiae is a useful model to study questions regarding cell aggregation, heterogeneity and cooperation. In this review, we discuss scenarios of group formation and how this promotes facultative multicellularity in S. cerevisiae. We first describe proximate mechanisms leading to aggregation. These mechanisms include staying together and coming together, and can lead to group heterogeneity. Heterogeneity is promoted by nutrient limitation, structured environments and aging. We then characterize the evolutionary benefits and costs of facultative multicellularity in yeast. We summarize current knowledge and focus on the newest state-of-the-art discoveries that will fuel future research programmes aiming to understand facultative microbial multicellularity.

341 Background: Brain metastases from esophageal cancer are extremely rare with an estimated incidence of 1.7% but limited survival. Some reports highlight the rich microvascular density of such tumors and their distinctive degree of HER2, HIF1-a and EGFR expression, further emphasizing the potential role of angiogenesis in their neurotropic behavior. With this in mind, we aimed to illustrate the transcriptomic landscape of brain metastases from esophageal cancers and better understand the disease biology. Methods: Following IRB approval, we collected retrospective data on patients with a diagnosis of esophageal cancer with histology-proven brain metastases treated at our institution between 2008 and 2020. We identified 10 adequate, paired samples with available tissue for RNA extraction. Expression data was generated using NanoString TS 360 panels to characterize gene signatures of interest and analyzed using GSEA_4.1.0. Results: All 10 patients were Caucasian, 90% were male. Median age was 63 years. All had adenocarcinoma with G2-G3 histology. Median follow-up was 39.4 mos (95%CI, 27.4 to 51.4). At data cutoff, 6 patients had died. Median OS was 24.6 mos. Median time from curative surgery to CNS recurrence was 8.3 mos. Ninety percent of patients developed brain metastases within 24 mos of surgery. Median time from brain metastases to death was 4.7 mos. Almost a third of patients had HER2 positive disease. 48 gene signatures were analyzed; 5 being significantly enriched between metastatic and primary site accounting for cell cycle, DNA damage repair, MYC, mTOR signaling and immortality and stemness. Out of a pool of 760, significant overlap was seen between 12 genes across the 5 signatures of interest: POLE, LIG1, FEN1 (involved in DNA repair and replication); AURKA and PLK1 (cell proliferation and triggers for G2/M transition), BUB1 (establishment of the mitotic spindle checkpoint and chromosome congression), MCM2 and MCM4 (initiation of genome replication); BRIP1 and RAD51 (critical in homologous recombinational repair); CCNE1 (cell cycle G1/S transition) and WRAP53 (dual role as p53 regulator and protein involved in telomere elongation and DNA repair). ERBB2 was similarly enriched in primary and metastatic sites. Conclusions: These preliminary data demonstrate that the genomic basis of brain metastases in esophageal cancer goes beyond EGFR/ERBB2 signaling. A complex genomic interaction is present in brain metastases as compared to primary site that offers these cells the advantage for neurotropism. To our knowledge this is the first study attempting to illustrate genomic differences between primary and matched brain metastasis sites in esophageal cancer.

Research on symbiotic communities (microbiomes) of multicellular organisms seems to be changing our understanding of how species of plants and animals have evolved over millions of years. The quintessence of these discoveries is the emergence of the hologenome theory of evolution, founded on the concept that a holobiont (a host along with all of its associated symbiotic microorganisms) acts a single unit of selection in the process of evolution. Although the hologenome theory has become very popular among certain scientific circles, its principles are still being debated. In this paper, we argue, firstly, that only a very small number of symbiotic microorganisms are sufficiently integrated into multicellular organisms to act in concert with them as units of selection, thus rendering claims that holobionts are units of selection invalid. Secondly, even though holobionts are not units of selection, they can still constitute genuine units from an evolutionary perspective, provided we accept certain constraints: mainly, they should be considered units of co-operation. Thirdly, we propose a reconciliation of the role of symbiotic microorganisms with the theory of speciation through the use of a developed framework. Mainly, we will argue that, in order to understand the role of microorganisms in the speciation of multicellular organisms, it is not necessary to consider holobionts units of selection; it is sufficient to consider them units of co-operation.

We currently perform open-heart procedures using bloodless priming of cardiopulmonary bypass circuits regardless of a patient’s body weight. This study presents results of this blood-saving approach in neonates and infants with a body weight of up to 7 kg. It tests with multivariate analysis factors that affect perioperative transfusion. A total of 498 open-heart procedures were carried out in the period 2014–2016 and were analysed. Priming volume ranged from 73 ml for patients weighing up to 2.5 kg to 110 ml for those weighing over 5 kg. Transfusion threshold during cardiopulmonary bypass was 8 g/dl of haemoglobin concentration. Transfusion factors were first analysed individually. Variables with a p-value lower than 0.2 underwent logistic regression. Extracorporeal circulation was conducted without transfusion of blood in 335 procedures – that is, 67% of cases. Transfusion-free operation was achieved in 136 patients (27%) and was more frequently observed after arterial switch operation and ventricular septal defect repair (12/18=66.7%). It was never observed after Norwood procedure (0/33=0%). Lower mortality score (p=0.001), anaesthesia provided by a certain physician (p=0.006), first chest entry (p=0.013), and higher haemoglobin concentration before going on bypass (p=0.013) supported transfusion-free operation. Early postoperative mortality was 4.4% (22/498). It was lower than expected (6.4%: 32/498). In conclusion, by adjusting the circuit, cardiopulmonary bypass could be conducted without donor blood in majority of patients, regardless of body weight. Transfusion-free open-heart surgery in neonates and infants requires team cooperation. It was more often achieved in procedures with lower mortality score.

Division of labour between different specialized cell types is a central part of how we describe complexity in multicellular organisms. However, it is increasingly being recognized that division of labour also plays an important role in the lives of predominantly unicellular organisms. Saccharomyces cerevisiae displays several phenotypes that could be considered a division of labour, including quiescence, apoptosis and biofilm formation, but they have not been explicitly treated as such. We discuss each of these examples, using a definition of division of labour that involves phenotypic variation between cells within a population, cooperation between cells performing different tasks and maximization of the inclusive fitness of all cells involved. We then propose future research directions and possible experimental tests using S. cerevisiae as a model organism for understanding the genetic mechanisms and selective pressures that can lead to the evolution of the very first stages of a division of labour. Copyright © 2017 John Wiley & Sons, Ltd.

We present own results of the treatment of acute ischemic stroke by mechanical thrombectomy. Procedures were conducted in the Center for Acute Ischemic Stroke Treatment (CITO) at the University Hospital in Krakow that was established for this study. The Center works in the 24/7 system, and according to the protocol 6 professionals who participate in the procedure starts it within 45 minutes after the call of the Center coordinator. Since January the 1st 2013 till September the 30th 2016, 96 procedures were performed. Three months followup was performed in the consecutive 74 patients (39 men), mean age: 65,8±13,1 and here we present their data. Mean NIHSS score on admission was 15,4±4,2 points. In 46 patients (62.2%) mechanical thrombectomy was preceded by rt-PA-IV administration. Mean time from stroke onset to groin puncture was 265±88,5 minutes. After the procedure TICI=3 was obtained by 26 patients (35.1%), 2b – by 12 patients (16.2%), 2a – 19 patients (25.7%), 1 – 8 patients (10.8%) and 0 – by 9 patients (12.2%). Head CT scan done 24 hours after thrombectomy did not show hemorrhagic transformation in 37 patients. According to ECASS1 classification HI1 was found in 11 patients (14.9%), H12 – in 16 patients (21.6%), PH1 – in 4 patients (5.4%) and PH2 – in 6 patients (8.1%). Fourteen patients (18.9%) died within 90 days after stroke onset, and 38 – scored 80-100 points in Barthel Index (51.4%). Presented results indicate similar safety and efficacy profile of mechanical thrombectomy in the treatment of ischemic stroke performed in the system that was organized for this specific reason.

Este artigo científico investiga como os estudos do jurista português José Joaquim Gomes Canotilho e do jurista alemão Robert Alexy podem contribuir para a efetivação dos Direitos Fundamentais previstos na atual Constituição da República Federativa do Brasil. Apesar de positivados da Constituição, a efetividade dos Direitos Fundamentais depende da interpretação e aplicação dada pelos operadores jurídicos, daí exsurge a importância da contribuição de ambos os autores, já que enfrentam os mesmos problemas na Constituição de seus países. Na primeira parte caracterizou-se a positivação dos Direitos Fundamentais na atual Constituição da República Federativa do Brasil. Na segunda tratou-se da aproximação do tema com o princípio democrático desenvolvido por Canotilho. Na terceira parte abordou-se a teoria dos Direitos Fundamentais de Alexy. Concluiu-se com a adaptação das principais ideias de ambos à realidade brasileira de modo a contribuir para a efetivação dos Direitos Fundamentais no país. A pesquisa justifica-se pela importância da temática para a contemporaneidade, mormente pela necessidade de efetivação dos Direitos Fundamentais. O trabalho foi desenvolvido sob método indutivo. A pesquisa foi bibliográfica e documental.

Introduction In-hospital incomplete and terminated at discharge myocardial revascularization has significant impact on mortality after acute myocardial infarction (AMI), also in patients (pts) with reduced left ventricle ejection fraction (LVEF) ≤35%. However, subjects with LVEF >35%, who are not candidates for implantable cardioverter defibrillators, are still at risk. Authors hypothesized, that in those pts, the prognosis could be related to completeness of revascularization. Purpose To evaluate the risk of death and major adverse cardiovascular events (MACE) among pts with AMI and LVEF>35% in relation to myocardial revascularization status. Methods Single center prospective study encompassed 445-pts with AMI and LVEF>35%, who were treated with percutaneous coronary intervention and who survived in-hospital period. Study population was divided into two groups: group 1. – 73-pts with in-hospital incomplete and terminated revascularization at discharge; group 2. – 372-pts with complete or incomplete revascularization, in whom scheduled procedures were planned and performed (either percutaneous or surgical). The incidence of death and MACE was compared between groups during mean follow-up of 47.5 months after AMI. MACE was defined as a composite of death, recurrent AMI, non-scheduled revascularization, acute heart failure, stroke. Independent predictors for death were identified with multivariate Cox-regression models and expressed as hazard ratio (HR) with 95% confidence interval (CI). Results Patients in group 1. had higher mortality rate than in group 2. (26.4% vs. 9.1%; p Conclusions After invasive treatment of AMI, the prognosis in patients with LVEF>35% is related to revascularization status. In-hospital incomplete and terminated revascularization at discharge is an independent risk factor for death in this population.

Increasing demand for food production requires improvements as well as openness to new sources of protein. Single Cell Protein derived from microbes has been intensively studied as a supplement for traditional sources of proteins, both for animal feed and direct human consumption. Food grade yeast, including Saccharomyces cerevisiae, has already been successfully used in industry. Here, we describe an artificial selection experiment that resulted in isolating Saccharomyces cerevisiae lines with a heritable phenotype characterized by significantly higher total protein content. Compared to the ancestral population, the average increase in protein content for all the evolved lines containing multiple-evolved clones was 5.4 g per 100g of dry mass (~15% of the total protein content). However, we also obtained specific clones with a total protein content increase of 9.3 g/100g dry mass (increase by 24.6%). Whole genome sequence analysis of mutations acquired by these clones allowed us to hypothesize about the role of the amino acid signaling pathway (SPS) disorders as a genetic base of the increased amino acid content. The proposed method is based on gradient fractionation of starved haploid yeast cells of different density, which reflects their physiological state regarding quiescence. We found a positive correlation in the fraction of non-quiescent cells in the starved populations and their amino acid content. Our method does not require any genetic modification. We believe that it can be successfully applied to other Saccharomyces sp. in order to increase the amino acid content stored in their cells. Beyond its implications for applied science, knowledge of the quiescent state in yeast is of fundamental importance to our understanding of the genetic basis of the G0 state in eukaryotic cells.

Pulmonary rehabilitation (PR) has been recommended as an integral part of treatment for patients with chronic obstructive pulmonary disease (COPD). Climate therapy in salt mine chambers has been found of benefit in chronic respiratory diseases. The study compares long-term effects of underground PR in the Wieliczka Salt Mine with that conducted on the surface. There were 42 COPD patients enrolled in the study, with FEV

Background We routinely start cardiopulmonary bypass (CPB) for pediatric congenital heart surgery without homologous blood, due to circuit miniaturization, and blood-saving measures. Blood transfusion is applied if hemoglobin concentration falls under 8 g/dL, or it is postponed to after coming off bypass or after operation. How this strategy impacts on postoperative mortality and morbidity, in infants weighing ≤ 7 kg? Methods Six-hundred fifteen open-heart procedures performed from January 2014 to June 2018 were selected. One-hundred sixty-three patients (26.5%) were transfused on CPB (group 1), while 452 (73.5%) patients were not transfused on CPB (group 2). Operative risk and complexity were similar in both groups. Postoperative mortality and morbidity were compared. Multiple logistic regression was used to detect factors independently associated with outcome. Results Observed mortality in nontransfused group (0.7% = 3/452) was significantly lower than expected (4.2% = 19/452): p = 0.0007, and much lower than in transfused group (6.7% = 11/163): p Conclusion The strategy of transfusion-free CPB course, feasible in most patients ≤ 7kg, was associated with improved outcome. Asanguineous priming of CPB circuit should become standard, even in neonates and infants.

Pulmonary rehabilitation (PR) has been recommended as an integral part of treatment for patients with chronic obstructive pulmonary disease (COPD). Climate therapy in salt mine chambers has been found of benefit in chronic respiratory diseases. The study compares long-term effects of underground PR in the Wieliczka Salt Mine with that conducted on the surface. There were 42 COPD patients enrolled in the study, with FEV1/FVC

Coevolution between different biological entities is considered an important evolutionary mechanism at all levels of biological organization. Here, we provide evidence for coevolution of a yeast killer strain (K) carrying cytoplasmic dsRNA viruses coding for anti-competitor toxins and an isogenic toxin-sensitive strain (S) during 500 generations of laboratory propagation. Signatures of coevolution developed at two levels. One of them was coadaptation of K and S. Killing ability of K first increased quickly and was followed by the rapid invasion of toxin-resistant mutants derived from S, after which killing ability declined. High killing ability was shown to be advantageous when sensitive cells were present but costly when they were absent. Toxin resistance evolved via a two-step process, presumably involving the fitness-enhancing loss of one chromosome followed by selection of a recessive resistant mutation on the haploid chromosome. The other level of coevolution occurred between cell and killer virus. By swapping the killer viruses between ancestral and evolved strains, we could demonstrate that changes observed in both host and virus were beneficial only when combined, suggesting that they involved reciprocal changes. Together, our results show that the yeast killer system shows a remarkable potential for rapid multiple-level coevolution.

Developing materials from renewable resources to produce nanocomposites with satisfactory mechanical properties is a vital progress in today's society, and meanwhile, the widespread use of these nanocomposites needs to be enabled through large-scale manufacturing. Herein, ultra-strong nanocomposites with an aligned structure that consisted of bio-based, biodegradable poly(lactic acid) (PLA) and cellulose nanocrystals are demonstrated, and these nanocomposites can be manufactured at a large scale through surface modification of the nanocrystals, liquid-assisted extrusion, and solid-state drawing. An ultimate strength of 353 MPa and a toughness of 107 MJ/m3 can be achieved by these nanocomposites, which are superior compared to the values of many other thermoplastic materials. The mechanism that explains the high toughness of these nanocomposites has been extensively investigated, revealing that sliding of the PLA crystallites present in the materials is the main factor. Moreover, these nanocomposites possess relatively high glass transition temperature and exhibit strain-responsive birefringence behavior, which indicates their great potential in not only structural applications, but also optical strain sensing areas.

Objective To estimate the economic burden to the health service of surgical site infection following caesarean section and to identify potential savings achievable through implementation of a surveillance programme.\ud Design Economic model to evaluate the costs and benefits of surveillance from community and hospital healthcare providers’ perspective.\ud Setting England.\ud Participants Women undergoing caesarean section in National Health Service\ud hospitals.\ud Main outcome measure Costs attributable to treatment and management of surgical site infection following caesarean section.\ud Results The costs (2010) for a hospital carrying out 800 caesarean sections a year based on infection risk of 9.6% were estimated at £18,914 (95% CI 11,521 to 29,499) with 28% accounted for by community care (£5,370). With inflation to 2019 prices, this equates to an estimated cost of £5.0m for all caesarean sections performed annually in England 2018-19, approximately £1,866 and £93 per infection managed in hospital and community respectively. The cost of surveillance for a hospital for one calendar quarter was estimated as £3,747 (2010 costs).\ud Modelling a decrease in risk of infection of 30, 20 or 10% between successive surveillance periods indicated that a variable intermittent surveillance strategy achieved higher or similar net savings than continuous surveillance. Breakeven was reached sooner with the variable surveillance strategy than continuous surveillance\ud when the baseline risk of infection was 10 or 15% and smaller loses with a baseline risk of 5%.\ud Conclusion Surveillance of surgical site infections after caesarean section with feedback of data to surgical teams offers a potentially effective means to reduce infection risk, improve patient experience and save money for the health service.\ud Strengths and limitations\ud • The model estimated both community (28%) and hospital costs (72%), providing a more representative estimate of overall economic burden to the health service.\ud • Time-matching of patients with and without infection according to length of post-operative stay provided a more accurate assessment of excess bed-days attributable to surgical site infection (2.6 days) than average excess length of stay (median difference 5 days) comparison by disentangling the impact of prolonged length of stay on increased chance of detecting an infection.\ud • Through capture and assessment of the costs and impact of surveillance, our model demonstrated the potential for savings through reductions in incidence of surgical site infections.\ud • Costs were obtained from NHS National Schedule Reference Costs and other sources rather than observed expenditure and assumptions made about the number of extra midwife and general practitioner appointments resulting from infection.\ud • The study was based on healthcare utilisation and did not assess direct and indirect costs borne by the patients or their carers.

Little is known about the prognostic role of fasting glucose after mechanical thrombectomy (MT).We investigated whether fasting glucose on the next day after MT was associated with long-term outcome in acute ischemic stroke patients according to diabetes.We retrospectively analyzed 181 consecutive patients with acute anterior circulation ischemic stroke who underwent MT in 2 comprehensive stroke centers in Poland. Glucose levels were evaluated on admission and on the next day after MT. Fasting hyperglycemia (FHG) was defined as the glucose level above 5.5 mmol/L. Unfavorable outcome was defined as modified Rankin scale (mRS) of 3-6 at day 90 from stroke onset.Patients with FHG had higher mRS at 3-month follow-up compared with those without FHG (3.71 ± 2.56 versus 1.87 ± 2.22, P.001). In the subgroup analyses, FHG was associated with poor neurological outcome in the group without diabetes (3.74 ± 2.52 versus 1.81 ± 3.74, P.001) but not with diabetes (3.64 ± 2.67 versus 2.30 ± 3.74, P= .11). Patients without diabetes who had FHG were older, had higher glucose on admission, higher prevalence of atrial fibrillation, cardioembolic stroke etiology and bleeding brain complications compared with the group with normal fasting glucose. After adjustment for potential confounders, fasting glucose (odds ratio [OR] 1.46; 95% CI 1.19-1.79, P.001), age (OR 1.06; 95% CI 1.02-1.10, P = .001), successful reperfusion (OR 0.09; 95% CI 0.04-0.22, P.001) and baseline NIHSS score (OR 1.18; 95% CI 1.08-1.29, P.001) were predictors of mRS 3-6 at 3-month follow-up in the whole group. In the subgroup without diabetes, fasting glucose (OR 1.57; 95% CI 1.17-2.11, P = .002), age (OR 1.05; 95% CI 1.01-1.08, P = .008), successful reperfusion (OR 0.11; 95% CI 0.04-0.30, P.001) and baseline NIHSS score (OR 1.14; 95% CI 1.04-1.26, P = .011) were independent predictors of unfavorable 3-month outcome.Fasting glucose on the next day after MT in patients with acute ischemic stroke is an independent risk factor for worse 3-month outcome.

Summary Background Since the launch of the national Surgical Site Infection (SSI) Surveillance Service in 1997, successive expansions of the programme provided hospitals with increasing flexibility in procedures to target through surveillance. Ensuring that the programme continues to meet hospitals' needs remains essential. Aim As a means to inform the future direction of the service, a survey of all acute National Health Service trusts was undertaken to assess and understand priorities for surveillance. Methods A web-based survey was circulated to acute NHS trust infection control teams in England, asking them to identify and rank (i) reasons for undertaking current SSI surveillance, (ii) priority surgical categories for future SSI surveillance, and (iii) reasons for prioritizing these categories. Findings Of the 161 trusts surveyed, 84 (52%) responded. Assessment of quality of care was identified as the most common driver for SSI surveillance activity. Considerable heterogeneity in priority areas was observed, with 24 different surgical categories selected as top priority. Of the procedures undertaken by 15 or more trusts, caesarean section (2.7), hip replacement (2.8) and coronary artery bypass graft (2.9) were highest ranked. All 17 categories in the current surveillance programme were selected as a top priority by one or more trusts. Conclusion Whereas the majority of hospitals' priorities for SSI surveillance are included in the current programme, the top-ranked priority, caesarean section, is not included. Given the diversity of priority areas, maintaining a comprehensive spectrum of categories in the national programme is essential to assist hospitals in addressing local priorities.

The studies were designed to determine the polyphenolic composition and biological activity of extracts from fruits (SFE) and leaves (SLE) of Saskatoon (Amelanchier alnifolia Nutt.) in relation to erythrocyte membranes. A detailed quantitative and qualitative analysis of extracts was conducted, using the chro- matographic (UPLC-DAD, UPLC-ESI-MS) and spectrophotometric (Folin-Ciocalteu) methods. The biological activity of the extracts was investigated in relation to erythrocytes and isolated membranes of erythrocytes by using spectrophotometric, fluorimetric and microscopic methods and determined on the basis of hemolytic and antioxidant activity of the extracts and their impact on physical properties of the membrane such as: osmotic resistance, shape of erythrocytes, packing order of the polar head of lipids and fluidity of the membrane. The results showed that the tested extracts are rich sources of polyphenols, primarily from the group of flavonoids; in leaves dominating flavonols and anthocyanins in fruits. The SFE and SLE extracts to varying degree modify the physical properties of the erythrocyte membrane, causing formation of echinocytes, an increase in osmotic resistance and changes in the polar part of the membrane. Furthermore, the substances markedly protect erythrocytes and their membranes against oxidation induced by different physico-chemical factors. The findings indicate that the polyphenolic compounds contained in extracts of Saskatoon do not destroy biological membranes but effectively protect them against oxidation by way of interacting with the membrane surface. The extracts could effectively protect the organism and food products from the harmful effects of free radicals.

Background: ACT proved to be a reliable tool in assessment of asthma control. Psychopathological factors can influence asthma control and the result of ACT. Aim: Analysis of psychopathological and personality factors on ACT outcomes. Methods: Asthma control was assessed using ACT among 104 stable asthmatics. Quality of life was assessed by Asthma Quality of Life Questionnaire (AQLQ). Patients were evaluated for psychopathologies using 28-item General Health Questionnaire (GHQ28). Personality traits were assessed with Eysenck’s Personality Inventory (EPI). Acceptance of asthma was measured by Acceptance of Illness Scale (AIS). Results: Patients with uncontrolled asthma (ACT score Conclusions: Uncontrolled asthma patients have more frequent psychopathologies estimated by GHQ28 and lower acceptance of their disease. However, they do not differ from controlled asthmatics in terms of personality traits.