Cellular and transcriptomic response to pathogenic and non-pathogenic Vibrio parahaemolyticus strains causing acute hepatopancreatic necrosis disease (AHPND) in Litopenaeus vannamei.

The shrimp industry has historically been affected by viral and bacterial diseases. One of the most recent emerging diseases is Acute Hepatopancreatic Necrosis Disease (AHPND), which causes severe mortality. Despite its significance to sanitation and economics, little is known about the molecular response of shrimp to this disease. Here, we present the cellular and transcriptomic responses of Litopenaeus vannamei exposed to two Vibrio parahaemolyticus strains for 98 h, wherein one is non-pathogenic (VpN) and the other causes AHPND (VpP). Exposure to the VpN strain resulted in minor alterations in hepatopancreas morphology, including reductions in the size of R and B cells and detachments of small epithelial cells from 72 h onwards. On the other hand, exposure to the VpP strain is characterized by acute detachment of epithelial cells from the hepatopancreatic tubules and infiltration of hemocytes in the inter-tubular spaces. At the end of exposure, RNA-Seq analysis revealed functional enrichment in biological processes, such as the toll3 receptor signaling pathway, apoptotic processes, and production of molecular mediators involved in the inflammatory response of shrimp exposed to VpN treatment. The biological processes identified in the VpP treatment include superoxide anion metabolism, innate immune response, antimicrobial humoral response, and toll3 receptor signaling pathway. Furthermore, KEGG enrichment analysis revealed metabolic pathways associated with survival, cell adhesion, and reactive oxygen species, among others, for shrimp exposed to VpP. Our study proves the differential immune responses to two strains of V. parahaemolyticus, one pathogenic and the other nonpathogenic, enlarges our knowledge on the evolution of AHPND in L. vannamei, and uncovers unique perspectives on establishing genomic resources that may function as a groundwork for detecting probable molecular markers linked to the immune system in shrimp.

Lytic activity of phages against bacterial pathogens infecting diabetic foot ulcers.

Complications of diabetes, such as diabetic foot ulcers (DFUs), are common, multifactorial in origin, and costly to treat. DFUs are the cause of nearly 90% of limb amputations among persons with diabetes. In most chronic infections such as DFU, biofilms are involved. Bacteria in biofilms are 100-1000 times more resistant to antibiotics than their planktonic counterparts. Multidrug-resistant (MDR) Staphylococcus aureus and Pseudomonas aeruginosa infections in DFUs may require alternative therapeutic agents such as bacteriophages ("phages"). This study describes the lytic activity of phage cocktails AB-SA01 (3-phage cocktail) and AB-PA01 (4-phage cocktail), which target S. aureus and P. aeruginosa, respectively. The host range and lytic effect of AB-SA01 and AB-PA01 on a planktonic culture, single-species biofilm, and mixed-species biofilm were evaluated. In vitro testing showed that 88.7% of S. aureus and 92.7% of P. aeruginosa isolates were susceptible to AB-SA01 and AB-PA01, respectively, in the planktonic state. The component phages of AB-SA01 and AB-PA01 infected 66% to 94.3% of the bacterial isolates tested. Furthermore, AB-SA01 and AB-PA01 treatment significantly (p < 0.05) reduced the biofilm biomass of their hosts, regardless of the antibiotic-resistant characteristics of the isolates and the presence of a non-susceptible host. In conclusion, the strong lytic activity, broad host range, and significant biofilm biomass reduction of AB-SA01 and AB-PA01 suggest the considerable potential of phages in treating antibiotic-resistant S. aureus and P. aeruginosa infections alone or as coinfections in DFUs.

Uninformed Teacher-Student for hard-samples distillation in weakly supervised mitosis localization.

BACKGROUND AND OBJECTIVE: Mitotic activity is a crucial biomarker for diagnosing and predicting outcomes for different types of cancers, particularly breast cancer. However, manual mitosis counting is challenging and time-consuming for pathologists, with moderate reproducibility due to biopsy slide size, low mitotic cell density, and pattern heterogeneity. In recent years, deep learning methods based on convolutional neural networks (CNNs) have been proposed to address these limitations. Nonetheless, these methods have been hampered by the available data labels, which usually consist only of the centroids of mitosis, and by the incoming noise from annotated hard negatives. As a result, complex algorithms with multiple stages are often required to refine the labels at the pixel level and reduce the number of false positives. METHODS: This article presents a novel weakly supervised approach for mitosis detection that utilizes only image-level labels on histological hematoxylin and eosin (H&E) images, avoiding the need for complex labeling scenarios. Also, an Uninformed Teacher-Student (UTS) pipeline is introduced to detect and distill hard samples by comparing weakly supervised localizations and the annotated centroids, using strong augmentations to enhance uncertainty. Additionally, an automatic proliferation score is proposed that mimicks the pathologist-annotated mitotic activity index (MAI). The proposed approach is evaluated on three publicly available datasets for mitosis detection on breast histology samples, and two datasets for mitotic activity counting in whole-slide images. RESULTS: The proposed framework achieves competitive performance with relevant prior literature in all the datasets used for evaluation without explicitly using the mitosis location information during training. This approach challenges previous methods that rely on strong mitosis location information and multiple stages to refine false positives. Furthermore, the proposed pipeline for hard-sample distillation demonstrates promising dataset-specific improvements. Concretely, when the annotation has not been thoroughly refined by multiple pathologists, the UTS model offers improvements of up to ∼4% in mitosis localization, thanks to the detection and distillation of uncertain cases. Concerning the mitosis counting task, the proposed automatic proliferation score shows a moderate positive correlation with the MAI annotated by pathologists at the biopsy level on two external datasets. CONCLUSIONS: The proposed Uninformed Teacher-Student pipeline leverages strong augmentations to distill uncertain samples and measure dissimilarities between predicted and annotated mitosis. Results demonstrate the feasibility of the weakly supervised approach and highlight its potential as an objective evaluation tool for tumor proliferation.

Optimizing in vitro phage-ciprofloxacin combination formulation for respiratory therapy of multi-drug resistant Pseudomonas aeruginosa infections.

Respiratory infection caused by multi-drug resistant (MDR) Pseudomonas aeruginosa is challenging to treat. In this study, we investigate the optimal dose of anti-pseudomonas phage PEV31 (103, 105, and 108 PFU/mL) combined with ciprofloxacin (ranging from 1/8× MIC to 8× MIC) to treat the MDR P. aeruginosa strain FADD1-PA001 using time-kill studies. We determined the impact of phage growth kinetics in the presence of ciprofloxacin through one-step growth analysis. Single treatments with either phage PEV31 or ciprofloxacin (except at 8× MIC) showed limited bactericidal efficiency, with bacterial regrowth observed at 48 h. The most effective treatments were PEV31 at multiplicity of infection (MOI) of 0.1 and 100 combined with ciprofloxacin at concentrations above 1× MIC, resulting in a >4 log10 reduction in bacterial counts. While the burst size of phage PEV31 was decreased with increasing ciprofloxacin concentration, robust antimicrobial effects were still maintained in the combination treatment. Aerosol samples collected from vibrating mesh nebulization of the combination formulation at phage MOI of 100 with 2× MIC effectively inhibited bacterial density. In summary, our combination treatments eradicated in vitro bacterial growth and sustained antimicrobial effects for 48 h. These results indicated the potential application of nebulization-based strategies for the combination treatment against MDR lung infections.

Effectiveness of a specific strength program of the gluteus maximus muscle to improve dynamic postural control in female basketball players. A randomized controlled trial.

BACKGROUND: Basketball is a team sport in which players perform multidirectional movements, jumps and landings, experiencing abrupt accelerations and decelerations and numerous changes of rhythm. In this sport, speed and intensity are two key factors that are associated with an increased risk of injury. The aim of this randomized controlled trial was to determine the effectiveness of a specific gluteus maximus strength programme as preventive work for young female basketball players, to improve dynamic postural stability and to observe its impact in the rate of lower limb injuries, vertical jump, dynamic knee valgus and pain. RESEARCH QUESTION: Is effective a strength programme to improve dynamic postural stability, vertical jump and dynamic valgus in female basketball players? METHODS: A hundred and thirteen female basketball players that play in professional clubs were recruited, reaching the final stage 92 (46 per group). One group (CG) received conventional injury prevention training while the experimental group (EG) added to the conventional team prevention program, a gluteus maximus strength programme of 5 months composed of 4 exercises/2 days per week/2 sets of 10 repetitions per leg. RESULTS: The total injury incidence decreased from 0.33 to 0.16 cases (control group pre=0.43 to post=0.14 cases, EG pre=022 to post=0.19). The EG improved overall (p = 0.000), posterior (p = 0.001), posteromedial (p = 0.001) and posterolateral (p = 0.000) dynamic stability of the right leg; anterior (p = 0.024), medial (p = 0.07) and posteromedial (p = 0.01) of the left leg. Both groups improved vertical jump (GC: p = 0.045 and GE: p = 0.000). There was no significant improvement in pain or valgus. SIGNIFICANCE: This strength programme is effective in improving dynamic stability especially of the dominant leg and jump height.

Emotional distress and self-care during the COVID-19 pandemic in women from an indigenous migrant cultural collective in Mexico City / Malestares emocionales y su autoatención durante la pandemia de COVID-19 en mujeres de un colectivo cultural indígena migrante residente en la Ciudad de México

Abstract Introduction During the coronavirus (SARS-Cov-2) pandemic, restrictive measures were implemented to reduce contagion. However, severely decreasing social interaction also negatively impacted the economy, particularly that of indigenous groups. Objective This article seeks to understand the emotional distress identified by a group of indigenous women residents, as well as their self-care practices, during the COVID-19 pandemic in Mexico City. Method A digital qualitative study was undertaken since the fieldwork was conducted in person and online, using various Internet platforms, which served as a field scenario, data collection tool and a means of continuous connection with subjects. Results Anecdotal records were obtained from the subjects, who identified categorizations in the collective organization of the indigenous group, which became a support network for mobilizing official material resources. Information was also obtained on the way the women engaged in the self-care of their emotional distress in a range of ways with a sense of immediacy, through physical, spiritual, herbal, and psychological resources. They observed how women managed to cope with their situation and continue caring for and supporting their families to enable them to get by, distinguishing between those who were providers and those who were dependent on another provider. Discussion and conclusion The pandemic, together with social restrictions, created stressful situations, causing various emotional problems among the indigenous collective. Nevertheless, their capacity for self-management and self-care enabled them to cope with these conditions in the midst of structural contexts of violence, poverty, and social exclusion.

Resumen Introducción Durante la pandemia del nuevo coronavirus (SARS-Cov-2) se instauraron diferentes medidas restrictivas con la finalidad de disminuir los contagios. Sin embargo, al reducir severamente las interacciones sociales también se produjo un impacto negativo en la economía, especialmente en los grupos indígenas. Objetivo Este artículo busca conocer los malestares emocionales identificados por un colectivo de mujeres indígenas residentes, así como sus prácticas de auto-atención, durante la pandemia por COVID-19 en la CDMX. Método Se desarrolló una investigación cualitativa digital ya que el trabajo de campo fue presencial y en línea, así como en diferentes plataformas de la red de internet, las cuales fungieron como escenario de campo, herramienta de recopilación de datos y un dispositivo de conexión constante con los informantes Resultados Se obtuvieron registros anecdóticos de las participantes, que identificaron: categorizaciones en la organización colectiva del grupo indígena, convirtiéndose en red de apoyo que movilizó recursos materiales oficiales; cómo las mujeres practicaron la auto-atención de sus malestares emocionales de manera variada y con un sentido de inmediatez, mediante recursos físicos, espirituales, herbolarios, psicológicos y el saber aguantarse para sobrellevar su situación, y continuar cuidando y apoyando a sus familias a salir adelante, diferenciando entre mujeres proveedoras y las dependientes de otro proveedor. Discusión y conclusión La pandemia junto con las restricciones sociales, generaron situaciones estresantes, desencadenando diversas problemáticas emocionales en el colectivo indígena, pero su capacidad de autogestión y autocuidado les permitió sobrellevar tales condiciones en medio de contextos estructurales de violencia, pobreza y exclusión social.

Annotation protocol and crowdsourcing multiple instance learning classification of skin histological images: The CR-AI4SkIN dataset.

Digital Pathology (DP) has experienced a significant growth in recent years and has become an essential tool for diagnosing and prognosis of tumors. The availability of Whole Slide Images (WSIs) and the implementation of Deep Learning (DL) algorithms have paved the way for the appearance of Artificial Intelligence (AI) systems that support the diagnosis process. These systems require extensive and varied data for their training to be successful. However, creating labeled datasets in histopathology is laborious and time-consuming. We have developed a crowdsourcing-multiple instance labeling/learning protocol that is applied to the creation and use of the CR-AI4SkIN dataset.2 CR-AI4SkIN contains 271 WSIs of 7 Cutaneous Spindle Cell (CSC) neoplasms with expert and non-expert labels at region and WSI levels. It is the first dataset of these types of neoplasms made available. The regions selected by the experts are used to learn an automatic extractor of Regions of Interest (ROIs) from WSIs. To produce the embedding of each WSI, the representations of patches within the ROIs are obtained using a contrastive learning method, and then combined. Finally, they are fed to a Gaussian process-based crowdsourcing classifier, which utilizes the noisy non-expert WSI labels. We validate our crowdsourcing-multiple instance learning method in the CR-AI4SkIN dataset, addressing a binary classification problem (malign vs. benign). The proposed method obtains an F1 score of 0.7911 on the test set, outperforming three widely used aggregation methods for crowdsourcing tasks. Furthermore, our crowdsourcing method also outperforms the supervised model with expert labels on the test set (F1-score = 0.6035). The promising results support the proposed crowdsourcing multiple instance learning annotation protocol. It also validates the automatic extraction of interest regions and the use of contrastive embedding and Gaussian process classification to perform crowdsourcing classification tasks.

Stability of bacteriophages in organic solvents for formulations.

Bacteriophages or phages used as an alternative therapy for treating multi-drug resistant infections require formulation consideration. Current strategies to produce phage formulations involving organic solvents are based on empirical practices without a good understanding of phage stability during formulation development. In this study, we investigated the effect of common formulation organic solvents (ethanol, isopropyl alcohol, tetrahydrofuran (THF) and dimethyl sulfoxide (DMSO)) on the stability of Pseudomonas aeruginosa-specific myovirus (PEV1, PEV20) and podovirus (PEV31) phages using biological assay, transmission electron microscopy (TEM) and scattering near field optical microscopy (SNOM). The three phages were mixed with the solvents at different concentrations (25%, 50%, and 75% (v/v)) for 20 min. All phages were fully viable in the organic solvents at 25% (v/v) showing negligible titre changes. At the higher solvent concentration of 50% (v/v), the myoviruses PEV1 and PEV20 remained relatively stable (titre loss 0.4-1.3 log10), whereas the podovirus PEV31 became less stable (titre loss 0.25-3.8 log10), depending on the solvent used. Increasing the solvent level to 75% (v/v) caused increased morphological changes in TEM and decreased viability as indicated by the titre loss (0.32-7.4 log10), with DMSO being the most phage-destabilising solvent. SNOM spectra showed differences in the signal intensity and peak positions in the amide I and amide II regions, revealing altered phage proteins by the solvents. In conclusion, the choice of the solvents for phage formulation depends on both the phages and solvent types. Our results showed (1) the phages are more stable in the alcohols than DMSO and THF, and (2) the myoviruses tend to be more stable than the podovirus in the solvents. Overall, a low to moderate (25-50 % v/v) level of organic solvents (except 50% THF) can be used in formulation of the phages without a substantial titre loss.

Engaging fathers to strengthen the impact of early home visitation on physical child abuse risk: Findings from the dads matter-HV randomized controlled trial.

BACKGROUND: Despite growing recognition of the importance of fathers in child abuse risk, the field of perinatal home visitation has only begun to consider fathers' roles in the implementation of such services. OBJECTIVES: This study examines the effectiveness of Dads Matter-HV ("DM-HV"), a father-inclusion enhancement to home visitation, and hypothesized mediators of impact. METHODS: A multisite cluster randomized controlled trial was conducted with 17 home visiting program teams serving 204 families across study conditions. Program supervisors and their teams were randomized to deliver home visiting services plus DM-HV enhanced services (intervention) or home visiting services alone (control). Data were collected at three time points: baseline, 4 months post-baseline immediately following the intervention, and 12 months post-baseline. We employed structural equation modeling to estimate the effect of the intervention on physical child abuse risk and to trace hypothesized mediators, including the quality of the father-worker relationship, parents' partner support and abuse, and the timing of service initiation. RESULTS: Results indicated that the DM-HV enhancement improved home visitor relationships with fathers, but only for families receiving services initiated postnatally. For these families, the improved quality of the father-worker relationship predicted improved parents' support of one another and reduced bidirectional mother-father partner abuse at 4-month follow-up, which in turn lowered maternal physical child abuse risk and paternal physical child abuse risk at 12-month follow-up. CONCLUSIONS: DM-HV can strengthen the impact of home visitation services on physical child abuse risk for families when services are initiated postnatally.

Lytic Bacteriophage Is a Promising Adjunct to Common Antibiotics across Cystic Fibrosis Clinical Strains and Culture Models of Pseudomonas aeruginosa Infection.

Bacteriophages (phages) are antimicrobials with resurgent interest that are being investigated for the treatment of antibiotic refractory infection, including for Pseudomonas aeruginosa (Pa) lung infection in cystic fibrosis (CF). In vitro work supports the use of this therapy in planktonic and biofilm culture models; however, consistent data are lacking for efficacy across different clinical Pa strains, culture models, and in combination with antibiotics in clinical use. We first examined the efficacy of a 4-phage cocktail as an adjunct to our CF centre's first-line systemic combination antibiotic therapy (ceftazidime + tobramycin) for 16 different clinical Pa strains and then determined subinhibitory interactions for a subset of these strains with each antibiotic in planktonic and biofilm culture. When a 4-phage cocktail (4 × 108 PFU/mL) was added to a ceftazidime-tobramycin combination (ceftazidime 16 mg/mL + tobramycin 8 mg/mL), we observed a 1.7-fold and 1.3-fold reduction in biofilm biomass and cell viability, respectively. The four most antibiotic resistant strains in biofilm were very susceptible to phage treatment. When subinhibitory concentrations of antibiotics and phages were investigated, we observed additivity/synergy as well as antagonism/inhibition of effect that varied across the clinical strains and culture model. In general, more additivity was seen with the phage-ceftazidime combination than with phage-tobramycin, particularly in biofilm culture, where no instances of additivity were seen when phages were combined with tobramycin. The fact that different bacterial strains were susceptible to phage treatment when compared to standard antibiotics is promising and these results may be relevant to ongoing clinical trials exploring the use of phages, in particular in the selection of subjects for clinical trials.

The Effect of Dads Matter-HV on Father Engagement in Home Visiting Services.

Engaging fathers early in child and family services has the potential to promote positive father contributions towards positive child development, improve family well-being, and enhance service outcomes over time. However, low father engagement in child and family services remains a persistent problem, and few interventions designed to improve father engagement in these services have been rigorously tested. The current study assesses the effect of a service enhancement intervention called Dads Matter-Home Visiting (Dads Matter-HV) on biological father engagement in home visiting services when compared to home visiting services delivered as usual. To assess the efficacy of the Dads Matter-HV intervention, the research team used a stratified cluster randomized clinical trial design with five agencies delivering early home visiting service programs. Seventeen teams across the five agencies were randomly assigned to either the control group condition (i.e., standard home visiting services as usual) or the intervention condition (i.e., Dads Matter-HV). Data were collected from a total of 204 families at baseline, 4 months postbaseline (92% retention rate), and 12 months postbaseline (84% retention rate). The results suggest that Dads Matter-HV increases biological father engagement for fathers who begin services in the postnatal period, but reduces engagement when services are initiated prenatally. Findings suggest some pathways through which the intervention effects engagement.

Evaluating the effect of curcumin on the metacestode of Taenia crassiceps.

Curcumin, a curcuminoid present in the rhizome of the plant Curcuma longa has multiple pharmacological effects including anticarcinogenic and anti-inflammatory properties. This work evaluates the anthelmintic effect of the curcumin molecule (98% pure) on Taenia crassiceps cysticerci viability in vitro. Cysticerci incubated in the presence of increasing concentrations of curcumin showed a dose-dependent mortality correlated with a significant increase in the production of reactive oxygen species and a partial inhibition of thioredoxin-glutathione reductase, the only disulfide reductase present in these parasites. At 500 µM curcumin, a 100% of cysticerci lethality was obtained after 2 h of treatment. These results suggest the curcumin-induced oxidative stress could be in the origin of the anthelminthic effect of curcumin. Mice with cysticerci were injected intraperitoneally with 20, 40, or 60 mM curcumin daily for 30 days. A decrease in the burden of cysticerci (46%) was observed with a 60 mM dose of curcumin, supporting this compound as a potential anthelmintic drug.

Topical liquid formulation of bacteriophages for metered-dose spray delivery.

Bacteriophage (phage) therapy is a promising treatment strategy to combat antibiotic-resistant bacteria. Clinical reports from a century ago, as well as recent reports have revealed safety and efficacy of phage therapy for bacterial wound infections. However, the conventional liquid phage formulation and delivery platforms reported lack of dose control as it easily runs off from the infection site and it is impossible to determine total volume transfer. The aim of this study was to formulate phage liquids for topical delivery using a metered-dose spray. Two types of anti-Pseudomonas phages, PEV1 (myovirus) and PEV31 (podovirus) were formulated in 35% ethanol in water containing non-ionic polymers. The formulations were evaluated for physical properties, ease of spray, dripping upon spraying, drying time, in vitro release profiles, antibacterial activity, and storage stability. The optimized phage-polymer spray formulations were easily sprayable with minimal dripping and fast drying time. Phages were rapidly released from the formulation and inhibited the growth of Pseudomonas aeruginosa. Both PEV1 and PEV31 remained biologically stable in the optimized formulations during storage at 4 °C for eight weeks. This study showed the topical spray formulations containing non-ionic polymers in ethanol/water could be a promising and innovative therapeutic system for delivering phages.

The effects of different doses of inhaled bacteriophage therapy for Pseudomonas aeruginosa pulmonary infections in mice.

OBJECTIVES: Inhaled phage therapy has been revisited as a potential treatment option for respiratory infections caused by multidrug-resistant Pseudomonas aeruginosa; however, there is a distinct gap in understanding the dose-response effect. The aim of this study was to investigate the dose-response effect of Pseudomonas-targeting phage PEV31 delivered by the pulmonary route in a mouse lung infection model. METHODS: Neutropenic BALB/c mice were infected with multidrug-resistant P. aeruginosa (2 × 104 colony-forming units) through the intratracheal route and then treated with PEV31 at three different doses of 7.5 × 104 (Group A), 5 × 106 (Group B), and 5 × 108 (Group C) plaque-forming units, or phosphate-buffered saline at 2 hours postinoculation. Mice (n = 5-7) were euthanized at 2 hours and 24 hours postinfection, and lungs, kidneys, spleen, liver, bronchoalveolar lavage fluid, and blood were collected for bacteria and phage enumeration. RESULTS: At 24 hours postinfection, all phage-treated groups exhibited a significant reduction in pulmonary bacterial load by 1.3-1.9 log10, independent of the delivered phage dose. The extent of phage replication was negatively correlated with the dose administered, with log10 titre increases of 6.2, 2.7, and 9 for Groups A, B, and C, respectively. Phage-resistant bacterial subpopulations in the lung homogenate samples harvested at 24 hours postinfection increased with the treatment dose (i.e. 30%, 74%, and 91% in respective Groups A-C). However, the mutants showed increased susceptibility to ciprofloxacin, impaired twitching motility, and reduced blue-green pigment production. The expression of the inflammatory cytokines (IL-1ß and IL-6, and TNF-α) was suppressed with increasing PEV31 treatment dose. DISCUSSION: This study provides the dose-response effect of inhaled phage therapy that may guide dose selection for treating P. aeruginosa respiratory infections in humans.

Enteric-coated bacteriophage tablets for oral administration against gastrointestinal infections.

Intestinal Pseudomonas aeruginosa is highly problematic in immunocompromised patients such as those in intensive care units in hospitals. Phage therapy is an attractive alternative or supplementary therapy to antibiotics as it not only kills multidrug-resistant bacteria, but also minimises the disruption of gut microflora. Solid oral dosage forms (i.e., tablets) have the potential to effectively deliver viable phages to the gastrointestinal tract, but formulation studies have been scarce. In this study, Pseudomonas-targeting phage PEV20 was used as a model to produce tablets suitable for oral delivery by utilising industry-scale tablet compression and tablet coating machines. Phage tablets were produced by (i) spray drying of phages, (ii) direct compression of the phage powders into tablets, and then (iii) tablet coating. The resulting phage tablets had negligible phage titre reduction throughout the process and passed the British Pharmacopeia tests, including friability, weight variation, disintegration and dissolution of the tablets as well as weight gain and disintegration (in 0.1 M HCl and pH 7.4 phosphate buffer) of coated tablets. The developed formulation method can be utilised to produce tablets containing other phages and phage cocktails that are effective against enteric bacterial infections.

The complete mitochondrial genome of the fine flounder Paralichthys adspersus revealed by next-generation sequencing.

The complete mitochondrial genome of the fine flounder Paralichthys adspersus, was determined for the first time through Next Generation Sequencing (NGS) approach. The mitogenome (GenBank accession no. MW288827) has 17,060 bp in length and consisted of the well-known 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and the control region. The overall nucleotide composition of the whole mitogenome was A: 27.5%, C: 29.5%, G: 17.1%, and T: 25.9%. Phylogenetic analyses based on 12 protein-coding genes clustered P. adspersus in the monophyletic Paralichthyidae clade, showing the closest phylogenetic relationship with its congeneric species P. olivaceus.

Hydrogel formulations containing non-ionic polymers for topical delivery of bacteriophages.

Hydrogel is an attractive delivery vehicle for phages as it keeps the wound moist, acts as a protective barrier and facilitates wound healing process. The aim of this study was to formulate biologically stable phage hydrogels that enable controlled release of infective phages. Pseudomonas-targeting phages, PEV1 (myovirus) and PEV31 (podovirus) were formulated in hydrogels (109 PFU/g) consisting of non-ionic polymers, including hydroxyethyl cellulose (HEC), hydroxypropyl methylcellulose (HPMC), polyethylene oxide (PEO), polyvinyl alcohol (PVA), hydroxypropyl cellulose (HPC) and polyvinylpyrrolidone (PVP). The formulations were evaluated for physical properties, in vitro release profiles, antibacterial activity, and storage stability. Controlled release of phages was observed in 7.5% PEO, 20% PVA and 75% PVP hydrogels with >108 PFU release within 8 h. Poor phage release (7 × 105-4 × 107 PFU) was observed in 5% HPMC, 5% HEC and 30% HPC gels. The biostability of the optimized hydrogels was phage-specific with less titer loss observed for PEV1 (0-0.8 log) than for PEV31 (0.3-1.4 log). Both phages remained stable in PEO, PVA and HPMC hydrogels with <1 log titer reductions when stored at 5 °C. This study showed that 7.5% PEO and 20% PVA hydrogel formulations could be promising therapeutic systems for delivering phages for the treatment of wound infections.

Overcoming bacteriophage insensitivity in Staphylococcus aureus using clindamycin and azithromycinat subinhibitory concentrations.

BACKGROUND: Staphylococcus aureus is a pathogen of major concern in both acute infections and chronic conditions such as chronic rhinosinusitis (CRS). Bacteriophage (phage) therapy has recently regained interest for its potential to treat infections caused by antibiotic resistant strains including Methicillin Resistant Staphylococcus aureus (MRSA). However, bacteria can adapt and become resistant to phages. The aim of this study is to determine the potential for antibiotics to overcome phage resistance. METHODS: The susceptibility of S. aureus clinical isolates (CIs) to phages J-Sa36, Sa83 and Sa87 alone or in combination with protein synthesis inhibitor (PSI) antibiotics clindamycin, azithromycin and erythromycin was assessed using plaque spot assays, minimum inhibitory concentration (MIC) assays, double layer spot assays and resazurin assays. The safety and efficacy of subinhibitory PSI antibiotics in combination with phage was tested in a Sprague Dawley rat model of sinusitis infected with a phage resistant S. aureus CI. RESULTS: All three antibiotics at subinhibitory concentrations showed synergy when combined with all 3 phages against S. aureus CIs in planktonic and biofilm form and could sensitize phage-resistant S. aureus to promote phage infection. The combination of topical subinhibitory clindamycin or azithromycin and phage was safe and could eradicate S. aureus sinonasal biofilms in vivo. CONCLUSION: Subinhibitory concentrations of PSI antibiotics could sensitize phage-resistant S. aureus and MRSA strains to phages in vitro and in vivo. This data supports the potential use of phage-PSI antibiotic combination therapies, in particular for difficult-to-treat infections with phage-resistant S. aureus and MRSA strains.

Contribution of Gray Matter Atrophy and White Matter Damage to Cognitive Impairment in Mildly Disabled Relapsing-Remitting Multiple Sclerosis Patients.

Cognitive impairment (CI) is frequently present in multiple sclerosis patients. Despite ongoing research, the neurological substrates have not been fully elucidated. In this study we investigated the contribution of gray and white matter in the CI observed in mildly disabled relapsing-remitting multiple sclerosis (RRMS) patients. For that purpose, 30 patients with RRMS (median EDSS = 2), and 30 age- and sex-matched healthy controls were studied. CI was assessed using the symbol digit modalities test (SDMT) and the memory alteration test. Brain magnetic resonance imaging, diffusion tensor imaging (DTI), voxel-based morphometry (VBM), brain segmentation, thalamic vertex analysis, and connectivity-based thalamic parcellation analyses were performed. RRMS patients scored significantly lower in both cognitive tests. In the patient group, significant atrophy in the thalami was observed. Multiple regression analyses revealed associations between SDMT scores and GM volume in both hemispheres in the temporal, parietal, frontal, and occipital lobes. The DTI results pointed to white matter damage in all thalamocortical connections, the corpus callosum, and several fasciculi. Multiple regression and correlation analyses suggested that in RRMS patients with mild disease, thalamic atrophy and thalamocortical connection damage may lead to slower cognitive processing. Furthermore, white matter damage at specific fasciculi may be related to episodic memory impairment.

Variability in Bacteriophage and Antibiotic Sensitivity in Serial Pseudomonas aeruginosa Isolates from Cystic Fibrosis Airway Cultures over 12 Months.

Antibiotic treatment for Pseudomonas aeruginosa (Pa) in cystic fibrosis is limited in efficacy and may lead to multi-drug resistance (MDR). Alternatives such as bacteriophages are being explored but well designed, and controlled trials are crucial. The rational selection of patients with bacteriophage susceptible infections is required for both safety and efficacy monitoring. We questioned whether bacteriophage susceptibility profiles were constant or variable over time, variability having been reported with antibiotics. Serial Pa isolates (n = 102) from 24 chronically infected cystic fibrosis (CF) patients over one year were investigated with plaque and antibiotic disc diffusion assays. Variable number tandem repeat (VNTR) analysis identified those patients with >1 isolate. A median (range) of 4 (3-6) isolates/patient were studied. Twenty-one (87.5%) individuals had a single VNTR type; three (12.5%) had two VNTR types at different times. Seventy-five percent of isolates were sensitive to bacteriophage at ≥ 1 concentration; 50% of isolates were antibiotic multidrug resistant. Serial isolates, even when representing a single VNTR type, varied in sensitivity to both bacteriophages and antibiotics. The rates of sensitivity to bacteriophage supports the development of this therapy; however, the variability in response has implications for the selection of patients in future trials which must be on the basis of current, not past, isolate testing.