Comparison of Outcomes in Patients With Muscle-invasive Bladder Cancer Treated With Radical Cystectomy Versus Bladder Preservation

Purpose: Radical cystectomy currently remains the standard of care for muscle-invasive bladder cancer. However, surgery can be associated with considerable morbidity and mortality, including the removal of the bladder. An alternative strategy is to preserve the bladder through concurrent chemoradiation following a maximal transurethral resection of the tumor. National protocols using a bladder-preservation approach have demonstrated disease-specific outcomes comparable to radical cystectomy in selected patients, but these results have not been replicated in previously reported population-based series. Here, we describe an outcomes analysis of patients with muscle-invasive bladder cancer treated with either radical surgery or bladder-preserving chemoradiation (BPCRT) for those patients meeting BPCRT criterion using the National Cancer Database (NCDB). Materials and Methods: Using the NCDB, patients with American Joint Commission on Cancer clinical T2-3, N0, M0 urothelial carcinoma diagnosed between 2004 and 2013 were included for analysis. Only patients treated with definitive intent with either radical cystectomy or concurrent chemotherapy and radiation after a maximal transurethral tumor resection were included. Propensity-score matching was used. Results: Among 8454 eligible patients, 7276 (86%) underwent radical cystectomy, and 1178 (14%) underwent BPCRT. Patients undergoing BPCRT were significantly older (median age, 77 vs. 68 y; P

https://ift.tt/2K3cNka

Chondrogenic Differentiation Processes in Human Bone-Marrow Aspirates Seeded in Three-Dimensional-Woven Poly(ɛ-Caprolactone) Scaffolds Enhanced by Recombinant Adeno-Associated Virus–Mediated SOX9 Gene Transfer

Human Gene Therapy, Ahead of Print.


https://ift.tt/2t4sKwO

Airway Basal Cells Are the Key for Cystic Fibrosis Gene Therapy

Human Gene Therapy, Volume 29, Issue 6, Page 641-642, June 2018.


https://ift.tt/2M0d5pF

Immune Gene Therapy and the International Conference on Lymphocyte Engineering (ICLE 2018)

Human Gene Therapy, Volume 29, Issue 6, Page vii-ix, June 2018.


https://ift.tt/2JJtMcf

In-vitro effects of the FS50 protein from salivary glands of Xenopsylla cheopis on voltage-gated sodium channel activity and motility of MDA-MB-231 human breast cancer cells

Voltage-gated sodium channel activity enhances the motility and oncogene expression of metastasic cancer cells that express a neonatal alternatively spliced form of the NaV1.5 isoform. We reported previously that FS50, a salivary protein from Xenopsylla cheopis, showed inhibitory activity against the NaV1.5 channel when assayed in HEK 293T cells and antiarrhythmia effects on rats and monkeys after induction of arrhythmia by BaCl2. This study aims to identify the effect of FS50 on voltage-gated sodium channel activity and the motility of MDA-MB-231 human breast cancer cells in vitro. NaV1.5 was abnormally expressed in the highly metastatic breast cancer cell line MDA-MB-231, but not in the MCF-7 cell line. FS50 significantly inhibited sodium current, migration, and invasion in a dose-dependent manner, but had no effect on the proliferation of MDA-MB-231 cells at the working concentrations (1.5–12 μmol/l) after a long-term treatment for 48 h. Meanwhile, FS50 decreased NaV1.5 mRNA expression without altering the total protein level in MDA-MB-231 cells. Correspondingly, the results also showed that MMP-9 activity and the ratio of MMP-9 mRNA to TIMP-1 mRNA were markedly decreased by FS50. Taken together, our findings highlighted for the first time an inhibitory effect of a salivary protein from a blood-feeding arthropod on breast cancer cells through the NaV1.5 channel. Furthermore, this study provided a new candidate leading molecule against antitumor cells expressing NaV1.5. Correspondence to Xueqing Xu, PhD, Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, People's Republic of China Tel: +86 206 164 8537; fax: +86 206 164 8655; e-mail: xu2003@smu.edu.cn Correspondence to Jiguo Wu, PhD, Guangdong Provincial Key Laboratory of Tropical Medicine, Department of Environmental Health, School of Public Health, Southern Medical University, Guangzhou 510515, People's Republic of China Tel: +86 206 164 8327; e-mail: allanjw@126.com Received February 12, 2018 Accepted May 27, 2018 Copyright © 2018 Wolters Kluwer Health, Inc. All rights reserved.

https://ift.tt/2LZR3mW

The influence of in vitro pectin fermentation on the human fecal microbiome

Pectin is a complex dietary fiber and a prebiotic. To investigate pectin-induced changes in the gut microbiome and their effects on the short chain fatty acids (SCFAs) production, we performed in vitro pectin ...

https://ift.tt/2t1iO7A

Development of a species-specific TaqMan-MGB real-time PCR assay to quantify Olsenella scatoligenes in pigs offered a chicory root-based diet

Olsenella scatoligenes is the only skatole-producing bacterium isolated from the pig gut. Skatole, produced from microbial degradation of l-tryptophan, is the main contributor to boar taint, an off-odor and off-f...

https://ift.tt/2JSItFK

Adaptations to marine versus terrestrial low temperature environments as revealed by comparative genomic analyses of the genus Psychrobacter

ABSTRACT

While cold-adapted bacteria isolated from marine or terrestrial low temperature environments share many similarities, cold-adapted bacteria from terrestrial environments usually grow over a broader range of temperatures suggesting different constraints of these two low temperature environments. The diversity of habitats from which Psychrobacter have been isolated (e.g. cold marine environments, frozen soils, permafrost and humans) provides a unique opportunity to examine habitat specific adaptations while reducing phylogenetic effects. Here, comparative genomic analyses of 26 strains of Psychrobacter revealed several clusters with characteristics that correlated with habitat. Marine and terrestrial Psychrobacter have amino acid composition typical of psychrophiles (e.g. fewer proline and lysine, more acidic) when compared to Psychrobacter strains associated with warm hosts, and have many potentially cold-adapted core genes (e.g. ClpX, DsbC, GroEL/GroES and MutS2). Marine and terrestrial Psychrobacter share many genes (e.g. FadB) not found in warm host Psychrobacter, which had their own distinct gene content (e.g. collagenase-like protease). Furthermore, terrestrial Psychrobacter were differentiated from marine Psychrobacter by the use of different cold adaptations and more hydrophobic and aliphatic proteins. These data suggest that terrestrial and marine Psychrobacter evolved from a mesophilic ancestor and are accumulating adaptations for low temperatures as well as for their respective habitats.

https://ift.tt/2ygqIyN

Prokaryotic assemblages in suspended and subglacial sediments within a glacierized catchment on Qeqertarsuaq (Disko Island), west Greenland

ABSTRACT

Microbes transported by glacial meltwater streams are thought to be a product of passive dispersal from both supra- and subglacial sources, though studies investigating the origins of these assemblages are scarce. Here, we conducted a survey within a large catchment containing multiple glaciers on Qeqertarsuaq (Disko Island), west Greenland, to investigate whether meltwater-exported microbial assemblages in suspended sediments differ between glacial meltwater streams, and if they reflect corresponding bulk subglacial and extraglacial sediment communities. Using 16S rRNA gene amplicon sequencing, we found proglacial stream assemblages substantially differ from one another, despite their close spatial proximity. Furthermore, proglacial stream assemblages were composed of greater proportions of Cyanobacteria compared to bulk subglacial sediment communities, dominated by Betaproteobacteria, demonstrating large contributions of meltwater and microbial cells from supraglacial habitats. Corresponding physico-chemical characteristics of meltwater suggest that streams draining smaller glaciers had more equal contributions of both supra- and subglacial inputs compared with the main catchment outlet, aligning with observed changes in assemblage structure, such as the decreased proportion of Cyanobacteria. These results suggest that glacier size and hydrological drainage systems may influence the structure of exported microbial assemblages, and collectively provide insights into their formation and fate in thiscurrent age of deglaciation.

https://ift.tt/2MuUoeC

Regulatory effects of Shewanella putrefaciens isolated from shrimp Penaeus orientalis on the virulence factors of Vibrio parahaemolyticus and evaluation of the role of quorum sensing in virulence factors regulation

Abstract

As an aquatic pathogen widely present in aquatic food, Vibrio parahaemolyticus causes outbreaks of gastroenteritis across the globe. Virulence factors of V. parahaemolyticus increases with the amount of spoilage in aquatic organisms including shrimp, but mechanisms regulating its virulence factors are not well understood. In this study, five spoilage bacteria isolated from shrimp were investigated for their regulatory effects on the virulence factors including haemolysin and biofilm of V. parahaemolyticus. Among these isolates, Shewanella putrefaciens induced haemolytic activity in V. parahaemolyticus in a time-dose-temperature-dependent manner and we found the main component responsible for this effect to be the supernatant or cell-free extract of S. putrefaciens. Total haemolytic activity, expression of the thermostable direct haemolysin gene tdh and biofilm production of V. parahaemolyticus were significantly up-regulated by S. putrefaciens, but also by deletion of quorum-sensing luxM or luxS gene of V. parahaemolyticus. However, this regulation by S. putrefaciens was significantly impaired by deletion of the luxM gene, but not by deletion of the luxS gene. Further study showed that S. putrefaciens exhibited a strong degradation ability on the signalling molecule acylated homoserine lactone (AHL) synthesised by the LuxM enzyme. This study revealed a novel virulence regulatory mechanism that S. putrefaciens can significantly increase the virulence factors of V. parahaemolyticus via interfering with the luxM- type quorum-sensing signalling pathway through its AHL-degradation ability.

https://ift.tt/2ykzo7k

Fungal community composition and diversity vary with soil depth and landscape position in a no-till wheat-based cropping system

ABSTRACT

Soil edaphic characteristics are major drivers of fungal communities, but there is a lack of information on how communities vary with soil depth and landscape position in no-till cropping systems. Eastern Washington is dominated by dryland wheat grown on a highly variable landscape with steep, rolling hills. High-throughput sequencing of fungal ITS1 amplicons was used to characterize fungal communities across soil depth profiles (0 to 100 cm from the soil surface) among distinct landscape positions and aspects across a no-till wheat field. Fungal communities were highly stratified with soil depth, where deeper depths harbored distinct fungal taxa and more variable, less diverse fungal communities. Fungal communities from deep soils harbored a greater portion of taxa inferred to have pathotrophic or symbiotrophic in addition to saprotrophic lifestyles. Co-occurrence networks of fungal taxa became smaller and denser as soil depth increased. In contrast, differences between fungal communities from north-facing and south-facing slopes were relatively minor, suggesting that plant host, tillage, and fertilizer may be stronger drivers of fungal communities than landscape position.

https://ift.tt/2JI05bf

The causes and consequences of variation in human cytokine production in health

Melanie Schirmer | Vinod Kumar | Mihai G Netea | Ramnik J Xavier

https://ift.tt/2LWY0oN

Isolated Polycystic Pancreatic Disease: A Very Rare Cause of Pancreatic Insufficiency

A44-year-old woman presented with a 6-month history of 4 to 6 loose stools daily. She denied abdominal pain, weight loss, or fatigue. Laboratory tests including stool calprotectine were normal. Upper and lower endoscopy and biopsy specimens taken were unremarkable. Her serum carotene level was 1.7 mcg/dL (normal range, 2.3–65.0 mcg/dL), fecal elastase concentration was 120 mg/g (normal level, >200 mg/g), and a stool study was positive for qualitative fat. An abdominal ultrasound showed multiple pancreatic cysts.

https://ift.tt/2lgqtKW

Multi-component intrinsic brain activities as a safe, alternative to cortical stimulation for sensori-motor mapping in neurosurgery

Functional brain mapping for the precise identification of the primary sensori-motor area (S1-M1) prior to epilepsy surgery is key for reducing the risk of postoperative functional deficits. This particular functional brain map was traditionally introduced by several methodologies, such as functional magnetic resonance imaging (fMRI) (Chakraborty et al., 2008), somatosensory evoked potentials (Dinner et al., 1986), and electrical cortical stimulation (ECS) (Ojemann et al., 1989, Awad et al., 1991, Branco et al., 2003).

https://ift.tt/2MA76bQ

High expression of tumor necrosis factor receptor 2 in tissue is associated with progression and prognosis of esophageal squamous cell carcinoma

In recent years, the role of TNFR2 has attracted much attention for its promotion role in several types of tumors. However, in ESCC, the clinical relevance of TNFR2 is still unknown. In this study, we detected TNFR2 expression in ESCC tissues using IHC. The χ2 test showed that TNFR2 was positively correlated with invasion depth, advanced clinical stage and low differentiation; survival analysis showed that TNFR2 was positively correlated with poor OS; univariate Cox regression analysis showed that clinical stage, lymph node involvement and invasion depth were all correlated with OS; multivariate Cox regression analysis showed that lymph node involvement and invasion depth were independent prognostic factors.

https://ift.tt/2tfQ7mi

Gum disease may be a key initiator of rheumatoid arthritis related autoimmunity

The results of the study demonstrate increased levels of gum disease, and disease-causing bacteria, in individuals at risk of rheumatoid arthritis (RA).

https://ift.tt/2JPXVSV

Tumor shrinkage during chemoradiation in locally advanced cervical cancer patients: prognostic significance, and impact for image-guided adaptive brachytherapy

The prognostic value of GTV and its evolution after chemoradiotherapy, assessed in 247 patients were examined first on initial staging MRI and then at time of brachytherapy. In multivariate analysis, GTV optimal volume reduction was independently associated with improved overall survival and local control (p<0.001). Patients with optimal GTV reduction had no significant benefit from dose escalation D90 CTVHR ≥80Gy (p>0.3) while patients without optimal GTV reduction may have (p<0.05).

https://ift.tt/2JHpJgg

Integration of research into endoscopic practice

I work in an institution in which research is valued, but you do not need to do it to maintain a successful practice. Nor do you need to publish to move up the administrative food chain. Why then do members of our Digestive Disease Institute (DDI) have 138 active research projects as of this writing and routinely publish 50 to 100 peer-reviewed articles yearly? Admittedly, these are not all endoscopy-driven research projects. They include studies from other components of the DDI (GI surgeons, oncologists, pathologists, and interventional radiologists as well as, in our shop, an endocrinologist who is the director of our Nutritional Center of Excellence).

https://ift.tt/2JHVW7s

ESRD and Hospice Care in the United States: Are Dialysis Patients Welcome?

According to the National Hospice and Palliative Care Organization, hospice care is a "model for quality, compassionate care for people facing life-limiting illness" and is focused on caring, not curing, with a philosophy "to support the physical, psychosocial, and spiritual needs of patients and their loved ones so that dignity and quality of life may persevere through death and the period of familial bereavement."1 Kidney failure is unquestionably a life-limiting illness, with 5-year mortality for those receiving dialysis about twice that of adults with cancer, congestive heart failure, and stroke.

https://ift.tt/2JHAL5k

The Role of Big Data in the Development and Evaluation of US Dialysis Care

Rapid growth in electronic communications and digitalization, combined with advances in data management, analysis, and storage, have led to an era of "Big Data." The Social Security Amendments of 1972 turned end-stage renal disease (ESRD) care into a single-payer system for most patients requiring dialysis in the United States. As a result, there are few areas of medicine that have been as influenced by Big Data as dialysis care, for which Medicare's large administrative data sets have had a central role in the evaluation and development of public policy for several decades.

https://ift.tt/2LSZqAv

Childhood Kidney Disease: A Troubling Prognosis?

The 11th annual World Kidney Day was celebrated on March 10, 2016, across the globe in an effort to better inform the general public, policymakers, and the medical community about the importance of childhood kidney disease and the growing recognition that the antecedents of adult kidney disease can begin in childhood. A recent study by Calderon-Margalit et al1 in the New England Journal of Medicine provides further insight into the long-term outcomes of children with a history of kidney disorders that were assessed as having resolved by late adolescence.

https://ift.tt/2JIUL7L

High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize (Zea mays L.)

Here, we present a microscale protocol for processing grain samples and for incorporating this microscale approach into a high-throughput analytical pipeline. This is a higher throughput adaptation of currently available protocols.

https://ift.tt/2yjqY00

Protrusion Force Microscopy: A Method to Quantify Forces Developed by Cell Protrusions

Here, we detail the experimental techniques used to evaluate the protrusion forces that podosomes apply on a compliant film, from the preparation of the film to the automated analysis of topographical images.

https://ift.tt/2t2Cp7k

Evaluation of Commercial-Off-The-Shelf Wrist Wearables to Estimate Stress on Students

A protocol to evaluate solutions based on commercial-off-the-shelf (COTS) wrist wearables to estimate stress in students is proposed. The protocol is carried out in two phases, an initial laboratory-based stress induction test, and a monitoring stage taking place in the classroom while the student is performing academic activities.

https://ift.tt/2LXZBKK

High Throughput SiRNA Screening for Chloropicrin and Hydrogen Fluoride-Induced Cornea Epithelial Cell Injury

High throughput small inhibitory RNA screening is an important tool that could help to more rapidly elucidate the molecular mechanisms of chemical cornea epithelial injury. Herein, we present the development and validation of exposure models and methods for the high throughput screening of hydrogen fluoride- and chloropicrin-induced cornea epithelial injury.

https://ift.tt/2tehJbI

The Establishment of a Lung Colonization Assay for Circulating Tumor Cell Visualization in Lung Tissues

An animal model is needed to decipher the role of circulating tumor cells (CTCs) in promoting lung colonization during cancer metastasis. Here, we established and successfully performed an in vivo assay to specifically test the requirement of polymeric fibronectin (polyFN) assembly on CTCs for lung colonization.

https://ift.tt/2t2y3wW

Molecular heterogeneity and CXorf67 alterations in posterior fossa group A (PFA) ependymomas

Abstract

Of nine ependymoma molecular groups detected by DNA methylation profiling, the posterior fossa type A (PFA) is most prevalent. We used DNA methylation profiling to look for further molecular heterogeneity among 675 PFA ependymomas. Two major subgroups, PFA-1 and PFA-2, and nine minor subtypes were discovered. Transcriptome profiling suggested a distinct histogenesis for PFA-1 and PFA-2, but their clinical parameters were similar. In contrast, PFA subtypes differed with respect to age at diagnosis, gender ratio, outcome, and frequencies of genetic alterations. One subtype, PFA-1c, was enriched for 1q gain and had a relatively poor outcome, while patients with PFA-2c ependymomas showed an overall survival at 5 years of > 90%. Unlike other ependymomas, PFA-2c tumors express high levels of OTX2, a potential biomarker for this ependymoma subtype with a good prognosis. We also discovered recurrent mutations among PFA ependymomas. H3 K27M mutations were present in 4.2%, occurring only in PFA-1 tumors, and missense mutations in an uncharacterized gene, CXorf67, were found in 9.4% of PFA ependymomas, but not in other groups. We detected high levels of wildtype or mutant CXorf67 expression in all PFA subtypes except PFA-1f, which is enriched for H3 K27M mutations. PFA ependymomas are characterized by lack of H3 K27 trimethylation (H3 K27-me3), and we tested the hypothesis that CXorf67 binds to PRC2 and can modulate levels of H3 K27-me3. Immunoprecipitation/mass spectrometry detected EZH2, SUZ12, and EED, core components of the PRC2 complex, bound to CXorf67 in the Daoy cell line, which shows high levels of CXorf67 and no expression of H3 K27-me3. Enforced reduction of CXorf67 in Daoy cells restored H3 K27-me3 levels, while enforced expression of CXorf67 in HEK293T and neural stem cells reduced H3 K27-me3 levels. Our data suggest that heterogeneity among PFA ependymomas could have clinicopathologic utility and that CXorf67 may have a functional role in these tumors.

https://ift.tt/2tfh7lU

Timing of grip and goal activation during action perception: a priming study

Abstract

Several models of action recognition acknowledge the involvement of distinct grip and goal representations in the processing of others' actions. Yet, their functional role and temporal organization are still debated. The present priming study aimed at evaluating the relative timing of grip and goal activation during the processing of photographs of object-directed actions. Action could be correct or incorrect owing to grip and/or goal violations. Twenty-eight (Experiment 1) and 25 (Experiment 2) healthy adults judged the correctness of target actions according to object typical use. Target pictures were primed by action pictures sharing the same grip or same goal, both the same grip and same goal or none. Primes were presented for 66 or 300 ms in Experiment 1 and for 120 or 220 ms in Experiment 2. In Experiment 1, facilitative priming effects were observed for goal and grip similarity after 300 ms primes but only for goal after 66 ms primes. In Experiment 2, facilitative priming effects were found for both goal and grip similarity from 120 ms of prime processing. In addition, results from a control condition in Experiment 2 indicated that mere object priming could partially account for goal similarity priming effects, suggesting that object identity may help the observer to make predictions about possible action goals. Findings demonstrate an early and first activation of goal representations, as compared to grip representations, in action decoding, consistent with predictive accounts of action understanding. Future studies should determine to what extent the timing of grip and goal activation is context-sensitive.

https://ift.tt/2yhYxiR

Cancers, Vol. 10, Pages 206: Constitutive Activation of STAT3 in Myeloma Cells Cultured in a Three-Dimensional, Reconstructed Bone Marrow Model

Cancers, Vol. 10, Pages 206: Constitutive Activation of STAT3 in Myeloma Cells Cultured in a Three-Dimensional, Reconstructed Bone Marrow Model

Cancers doi: 10.3390/cancers10060206

Authors: Yung-Hsing Huang Ommoleila Molavi Abdulraheem Alshareef Moinul Haque Qian Wang Michael P. Chu Christopher P. Venner Irwindeep Sandhu Anthea C. Peters Afsaneh Lavasanifar Raymond Lai

Malignant cells cultured in three-dimensional (3D) models have been found to be phenotypically and biochemically different from their counterparts cultured conventionally. Since most of these studies employed solid tumor types, how 3D culture affects multiple myeloma (MM) cells is not well understood. Here, we compared MM cells (U266 and RPMI8226) in a 3D culture model with those in conventional culture. While the conventionally cultured cells were present in single cells or small clusters, MM-3D cells grew in large spheroids. We discovered that STAT3 was the pathway that was more activated in 3D in both cell lines. The active form of STAT3 (phospho-STAT3 or pSTAT3), which was absent in MM cells cultured conventionally, became detectable after 1&ndash;2 days in 3D culture. This elevated pSTAT3 level was dependent on the 3D environment, since it disappeared after transferring to conventional culture. STAT3 inhibition using a pharmacological agent, Stattic, significantly decreased the cell viability of MM cells and sensitized them to bortezomib in 3D culture. Using an oligonucleotide array, we found that 3D culture significantly increased the expression of several known STAT3 downstream genes implicated in oncogenesis. Since most primary MM tumors are naturally STAT3-active, studies of MM in 3D culture can generate results that are more representative of the disease.

https://ift.tt/2JQHFBb

Linear Chains of HER2 Receptors Found in the Plasma Membrane Using Liquid-Phase Electron Microscopy

The spatial distribution of the human epidermal growth factor 2 (HER2) receptor in the plasma membrane of SKBR3 and HCC1954 breast cancer cells was studied. The receptor was labeled with quantum dot (QD) nanoparticles, and fixed whole cells were imaged in their native liquid state with environmental scanning electron microscopy (ESEM) using scanning transmission electron microscopy (STEM) detection. The locations of individual HER2 positions were determined in a total plasma membrane area of 991 μm2 for several SKBR3 cells and 1062 μm2 for HCC1954 cells.

https://ift.tt/2LRs3hx

Maternity experiences of mothers with multiple disadvantages in England: A qualitative study

Publication date: Available online 15 June 2018
Source:Women and Birth
Author(s): Jenny McLeish, Maggie Redshaw
BackgroundDisadvantaged mothers and their babies are at increased risk of poor perinatal outcomes and have less positive experiences of maternity care.AimTo explore the maternity care experiences of mothers with multiple disadvantages.MethodsA qualitative descriptive study based on semi-structured interviews with 40 mothers with multiple disadvantages, using thematic analysis.FindingsFour themes emerged: 'A confusing and frightening time', 'Longing to be respected as an individual', 'The importance of choice and control', and 'Needing trust to feel safe'. Mothers brought feelings of powerlessness and low self-esteem to their encounters with maternity professionals, which could be significantly worsened by disrespectful care. They needed support to navigate the complex maternity system. Positive experiences were much more likely where the mother had received continuity of care from a specialist midwife or small team.Discussion and conclusionMothers with multiple disadvantages value being treated as an individual, making informed choices, and feeling safe, but they may lack the confidence to ask questions or challenge disrespectful treatment. Training and supervision should enable maternity professionals to understand how confusing maternity care can be to very disadvantaged mothers. It should emphasise the need to provide accessible and empowering information and guidance to enable all mothers to make choices and understand the system. Leaders of maternity services need to do more to challenge negative staff attitudes and ensure that that all mothers are treated at all times with kindness, respect and dignity. Specialist midwives can deliver a high quality service to mothers experiencing multiple disadvantages.



https://ift.tt/2tc4051

Scale-up of industrial microbial processes

Abstract

Scaling up industrial microbial processes for commercial production is a high-stakes endeavor, requiring time and investment often exceeding that for laboratory microbe and process development. Omissions, oversights and errors can be costly, even fatal to the program. Approached properly, scale-up can be executed successfully. Three guiding principles are provided as a basis: begin with the end in mind; be diligent in the details; prepare for the unexpected. A detailed roadmap builds on these principles. There is a special emphasis on the fermentation step, which is usually the costliest and also impacts downstream processing. Examples of common scale-up mistakes and the recommended approaches are given. It is advised that engineering resources skilled in integrated process development and scale-up be engaged from the very beginning of microbe and process development to guide ongoing R&D, thus ensuring a smooth and profitable path to the large-scale commercial end.

https://ift.tt/2tcVwLe

mRNA transcript distribution bias between Borrelia burgdorferi bacteria and their outer membrane vesicles

Abstract

Borrelia burgdorferi, a bacterium in the spirochete phylum, is the causative agent of Lyme disease. Borrelia burgdorferi has a linear chromosome with a number of circular and linear plasmids. Bacteria, including B. burgdorferi, release spherical outer membrane vesicles (OMVs) that are known to carry secretory products including metabolites, nucleic acids and proteins. Herein, we provide the first comparative transcriptomic analysis of the vesicles released from B. burgdorferi. We identified a total of ∼1200 unique transcripts with at least one mapped read from the bacterial cell and its OMVs. We compared the spectrum of transcripts between bacterial cell and its OMVs, and found a biased distribution based on the source of transcripts, i.e. plasmid-encoded transcripts are more likely to be enriched in the OMVs. We validated the distribution for some of the transcripts by qPCR. This analysis provides the first evidence that some of the B. burgdorferi transcripts are preferentially packaged in OMV, which further suggest that the bacteria might use its OMVs for bacteria–bacteria or bacteria–host communications. This report also suggests a possible involvement of Borrelia-derived OMVs in the development of Lyme disease in both early and post disease syndromes.

https://ift.tt/2Mvz2xC

Ciprofloxacin binding to GyrA causes global changes in the proteome of Pseudomonas aeruginosa

Abstract

Ciprofloxacin is one of the most widely-used antibiotics, and has proven especially effective at controlling infections associated with the opportunistic human pathogen, Pseudomonas aeruginosa. In this work, we show that sub-inhibitory concentrations of ciprofloxacin induce discrete changes in the intracellular proteome. Central metabolism and cell envelope-associated functions are particularly affected. In spite of the low magnitude of the intracellular proteomic changes, we found that sub-lethal concentrations of ciprofloxacin had substantial effects on motility and exoprotein secretion. Crucially, the proteomic and phenotypic modulations that we observed were absolutely dependent upon the presence of wild-type GyrA; an isogenic strain of P. aeruginosa carrying a ciprofloxacin-insensitive form of GyrA (a T83→I mutant) did not display ciprofloxacin-dependent changes unless complemented with wild-type gyrA in trans. These results show that the diverse effects of sub-inhibitory ciprofloxacin on the cell are routed through its primary target in the cell, DNA gyrase.

https://ift.tt/2JRGjq3

Antimicrobial activity of fusidic acid in Escherichia coli is dependent on the relative levels of ribosome recycling factor and elongation factor G

Abstract

During protein synthesis, elongation factor G (EFG) participates at the steps of translocation and ribosome recycling. Fusidic acid (FA) is a bacteriostatic antibiotic, which traps EFG on ribosomes, stalling them on mRNAs. How the bacterial susceptibility to FA is determined, and which of the two functions of EFG (translocation or ribosome recycling) is more vulnerable, has remained debatable. The in vivo studies addressing these aspects of FA mediated inhibition of protein synthesis are lacking. Here, we used a system of Escherichia coli strains and their complementation/supplementation with the plasmid borne copies of the inducible versions of EFG and ribosome recycling factor (RRF) genes. Additionally, we investigated FA sensitivity in a strain with increased proportion of stalled ribosomes. We show that the cells with high EFG/RRF (or low RRF/EFG) ratios are more susceptible to FA than those with low EFG/RRF (or high RRF/EFG) ratios. Our in vivo observations are consistent with the recent in vitro reports of dependence of FA susceptibility on EFG/RRF ratios, and the notion that an overriding target of FA is the translocation function of EFG. An applied outcome of our in vivo study is that FA mediated growth inhibition could be facilitated by depletion or inactivation of cellular RRF.

https://ift.tt/2tdYVJF

Antifungal activity of selected natural preservatives against the foodborne molds Penicillium verrucosum and Aspergillus westerdijkiae

Abstract

The present study examines the inhibitory effect of the essential oil (EO) of Origanum vulgare, its active components carvacrol and thymol, and a few active components of other EOs, namely, eugenol, trans-cinnamaldehyde and 1,8-cineole, against Penicillium verrucosum CBS 302.48, Aspergillus westerdijkiae CBS 112803 and Aspergillus westerdijkiae CBS 112804. Therefore, the minimum inhibitory concentration (MIC) was determined by broth macrodilution of each antifungal agent. Regarding their antifungal activity, the following ranking in order of decreasing inhibitory action is: trans-cinnamaldehyde > carvacrol = thymol > O. vulgare EO > eugenol > 1,8-cineole. The combined effects of the natural preservatives were studied by checkerboard assay. The results are based on the fractional inhibitory concentration indices (FICIs), which are interpreted as 'synergy', 'no interaction' or 'antagonism'. The indices ranging from 0.8 to 1.3 and thus show no interaction. In addition, the dependence of the natural inhibitors on different pH values (pH 7.0, 5.6, 4.5 and 3.5) and water activity (0.99, 0.92, 0.90 and 0.87 a_w) was investigated. All tested natural preservatives are tolerant to the different examined milieu conditions, with the lowest MICs recorded at pH 3.5 and 0.87 a_w.

https://ift.tt/2Mt4Raq

Photoirradiated caffeic acid as an antimicrobial treatment for fresh produce

Abstract

The antimicrobial efficacy of 400 nm photoirradiated caffeic acid (CA, 5 mM) was evaluated against Escherichia coli O157:H7 and Listeria innocua. A stronger antimicrobial effect was observed on E. coli than on L. innocua where the combined treatment resulted in 4 and 1 log(CFU/mL) reductions, respectively. The treatment's effects on cellular metabolism (resazurin assay), uptake of CA (fluorescence technique) and membrane damage (propidium iodide assay) were studied in both species. CA uptake increased in both species, but membrane damage was only observed in E. coli O157:H7. The treatment had minimal impact on metabolic activity in both species. The treatment applied to the surface of spinach leaves was found to be effective against E. coli O157:H7. The novel treatment proposed in this study has the potential to improve the microbial food safety of fresh produce.

https://ift.tt/2MwzeNm