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The discovery of the resoruclins, new 3,5-dihydroxybenzoic acid containing macrolides for Steptomyces sp. MST-91080, and their putative biosynthetic pathway.

The discovery of a novel family of p-nitrobenzoylated piperazines from Aspergillus brevijanus revealed that the biosynthesis of p-nitrobenzoic acid in fungi is catalysed by a PABA synthase, an aminodeoxychorismate synthase and a cytochrome P450.

While the phenomenon of beat-deafness has been explored in clinical contexts, few studies have investigated how rhythmically challenged people experience the act of keeping time with music. Task-based semi-structured interviews were conducted with eight participants who self-identified as being unable to clap in time to a beat. Participants were asked to keep time with a gradated battery of musical stimuli using both claps and alternative gestures (e.g. head nodding, swaying) and to articulate their timekeeping experience and strategies. Analysis reveals three core themes: (a) an apparent disconnect between the act of identifying the beat and the physical act of clapping, (b) variation in strategies for keeping time depending on the type of musical example (i.e., whether the beat was explicitly played by a percussive instrument or not), and (c) variation in the ease of coordination and listening experience when a movement other than clapping was used to keep time. Despite being small in scale, this qualitative study sheds light on some of the underlying strategies and processes involved in beat abstraction and keeping time to music, informing options for the musical training of rhythmically challenged people.

Background Despite more than 2 years having elapsed since the onset of SARS-CoV-2 pandemic, a level of hesitation around increased SARS-CoV-2 vaccine toxicity in cancer patients receiving immunotherapy (IO) remains. Here, we explore serological responses to SARS-CoV-2 vaccination in patients treated with IO and we describe blood cytokines, autoantibody levels and immune-related adverse events (irAEs) post vaccination. Methods Serum anti-SARS-CoV-2 spike (S) protein receptor binding domain (RBD) antibodies, surrogate viral neutralization test (sVNT), Th1/Th2 cytokines and antibodies against self-antigens were quantified at baseline, between 1st and 2nd vaccine doses, at 1 week (1W), 1 month (1M), 4–6 months and 10–12 months after the 2nd dose. Grade 2 or higher (≥ gr2+) irAEs were captured prospectively. Results Fifty-one evaluable patients were enrolled in this longitudinal study, 35 on immunotherapy (IO) and 16 on non-immunotherapy (non-IO) treatment. Absolute levels and neutralization potential of anti-SARS-CoV-2 antibodies were not significantly different in the IO group compared to non-IO. Chemotherapy adversely affects seroconversion when compared to IO and/or targeted treatment with antibody levels of 67.6 U/mL vs 1441 U/mL (p = 0.006) and sVNT of 70.9% vs 94.5% (p = 0.009), at 1M after 2nd vaccine dose. Following vaccination, the prevalence of grade ≥ 2 irAEs in patients treated with IO was not higher than the usual reported IO toxicity. We report, for the first time, that post-vaccination, IgM autoantibodies against beta 2 glycoprotein (p = 0.02), myeloperoxidase (p = 0.03), nucleosome (p = 0.041), SPLUNC2 (p

The molecular underpinnings of organ dysfunction in acute COVID-19 and its potential long-term sequelae are under intense investigation. To shed light on these in the context of liver function, we performed single-nucleus RNA-seq and spatial transcriptomic profiling of livers from 17 COVID-19 decedents. We identified hepatocytes positive for SARS-CoV-2 RNA with an expression phenotype resembling infected lung epithelial cells. Integrated analysis and comparisons with healthy controls revealed extensive changes in the cellular composition and expression states in COVID-19 liver, reflecting hepatocellular injury, ductular reaction, pathologic vascular expansion, and fibrogenesis. We also observed Kupffer cell proliferation and erythrocyte progenitors for the first time in a human liver single-cell atlas, resembling similar responses in liver injury in mice and in sepsis, respectively. Despite the absence of a clinical acute liver injury phenotype, endothelial cell composition was dramatically impacted in COVID-19, concomitantly with extensive alterations and profibrogenic activation of reactive cholangiocytes and mesenchymal cells. Our atlas provides novel insights into liver physiology and pathology in COVID-19 and forms a foundational resource for its investigation and understanding.

Background Skin diseases in cattle pose significant animal welfare issues and financial burdens. Microbial communities inhabiting the skin have essential roles in maintaining host health. Microbiota manipulation can be an efficient strategy for improving the productivity and sustainability of skin disease management. However, a lack of information on the skin microbiota of healthy cattle and how environmental and host factors drive its normal variation has limited using skin microbes for diagnosing or treating skin disease and pest infestation in cattle. Here, we profiled the skin microbiota of 1,734 healthy cattle from 25 different herds located in a 187,000 km2 area in the northeast to east Australia using 16S rRNA gene amplicon sequencing. The impact of host and climatic conditions on the skin microbial populations was examined. Results Our results demonstrate a highly diverse microbiota on the skin of healthy cattle. While the structure and diversity of microbial communities varied between herds, several bacteria were present consistently despite the differences in environmental conditions. For example, bacterial families Moraxellaceae, Sphingomonadaceae, Bacillaceae and Burkholderiaceae were observed in most cattle, however, their relative abundance varied significantly between herds. Solar exposure and rainfall were key factors contributing to the observed variations in skin microbiota between herds, while temperature and cattle breed did not show any statistically significant impact on the composition of cattle skin microbiota. Conclusions This is the first report of the composition of the commensal skin microbiota of healthy cattle, specifically herds located in the northeast to east Australia and the impact of environmental and host variables on these microbial communities. Our study provides valuable insight into normal variation in cattle skin microbiota, an essential step for developing skin microbiota-based therapies for skin disease present in cattle.

Amycolatopsis sp. MST-135876 was isolated from soil collected from the riverbank of El Pont de Suert, Catalonia, Spain. Cultivation of MST-135876 on a range of media led to the discovery of a previously unreported dichlorinated cyclic hexapeptide, suertide A (d-Ser, 5-Cl-d-Trp, 6-Cl-d-Trp, l-Ile, d-Val, d-Glu), featuring an unprecedented pair of adjacent 5/6-chlorotryptophan residues. Supplementing the growth medium with KBr resulted in production of the mono- and dibrominated analogues suertides B and C, respectively. Suertides A–C displayed selective activity against Bacillus subtilis (MIC 1.6 µg ml−1) and Staphylococcus aureus (MIC 3.1, 6.3, and 12.5 µg ml−1, respectively), while suertides A and B showed appreciable activity against methicillin-resistant S. aureus (MIC 1.6 and 6.3 µg ml−1, respectively).

This paper will explore how a synesthete, RC, perceives colors, shapes, and textures in silence. RC has sound-to-color synesthesia, where at least one color is associated with each pitch on the Western music scale. Listening to silence strips all sound down to the bare minimum in terms of color or texture. As discovered through weekly meditation in a “Deep Listening” class, RC places a pitch to everything at least audible when plunged into silence, trying hard to capture the colors of every available sound. The silence also forces a more pronounced tactile sense that RC tries to grasp together with the colors. Every experience with the sound however, is present but muddled, and difficult to understand. Silence becomes an uncomfortable world of uncertainty and it becomes vital to grasp the visual and tactile nuances that are a part of it. This paper reflects RC’s progression through silence towards an understanding of her senses. What is silence? And how does the synesthete grapple with it?

Purpose: To report a case of local metastasis of conjunctival melanoma, which may occur via extravascular migratory metastasis (EVMM), and discuss its clinical relevance in conjunctival melanoma tumor staging and possible management implications. Methods: Retrospective chart review of a single clinical case with clinicopathologic correlation. Results: A 65-year-old male referred due to local recurrence of conjunctival melanoma at the caruncle was successfully treated after two excisional procedures with negative sentinel lymph node biopsies. Forty-eight months after initial presentation, the patient developed a nodular lesion representing local recurrence in the ipsilateral upper tarsal conjunctiva, distant from the primary tumor site. Histopathology showed nodules in the substantia propria in the absence of primary acquired melanosis. The tumor cells were found along the extravascular surface without intralymphatic or intravascular tumor cells consistent with local metastasis. One possible mechanism is angiotropic microsatellitosis leading to local EVMM. Additional neck CT imaging showed no lymphadenopathy. Conclusion: EVMM via angiotropic microsatellitosis is another possible mechanism of noncontiguous local recurrence of conjunctival melanoma. Angiotropic microsatellitosis may represent a high-risk finding possibly related to increased melanoma-related mortality.

Since CRISPR/Cas9 was harnessed to edit DNA, the field of gene therapy has witnessed great advances in gene editing. New avenues were created for the treatment of diseases such as Cystic Fibrosis (CF). CF is caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. Despite the success of gene editing with the CRISPR/Cas9 in vitro, challenges still exist when using CRISPR/Cas9 in vivo to cure CF lung disease. The delivery of CRISPR/Cas9 into lungs, as well as the difficulty to achieve the efficiency required for clinical efficacy, has brought forth new challenges. Viral and non-viral vectors have been shown to deliver DNA successfully in vivo, but the sustained expression of CFTR was not adequate. Before the introduction of Helper-Dependent Adenoviral vectors (HD-Ad), clinical trials of treating pulmonary genetic diseases with first-generation viral vectors have shown limited efficacy. With the advantages of larger capacity and lower immunogenicity of HD-Ad, together with the versatility of the CRISPR/Cas9 system, delivering CRISPR/Cas9 to the airway with HD-Ad for lung gene therapy shows great potential. In this review, we discuss the status of the application of CRISPR/Cas9 in CF gene therapy, the existing challenges in the field, as well as new hurdles introduced by the presence of CRISPR/Cas9 in the lungs. Through the analysis of these challenges, we present the potential of CRISPR/Cas9-mediated lung gene therapy using HD-Ad vectors with Cystic Fibrosis lung disease as a model of therapy.

Early efforts in cystic fibrosis (CF) gene therapy faced major challenges in delivery efficiency and sustained therapeutic gene expression. Recent advancements in engineered site-specific endonucle...

Purpose To quantify the complete dimensions of the retro-orbicularis oculi fat (ROOF) pad and to determine its relationship to other fat compartments of the forehead. Methods The entire forehead of 14 hemifaces of seven fresh frozen human cadavers (four female, three male) was dissected in the subcutaneous and submuscular planes. For each plane, a ruler was placed at the facial midline, and images of the dissection plane were taken at 90° and 45°. Images were analyzed for vertical height, horizontal length, the distance to midline from the point of maximal height, and area for each hemiface of the ROOF and for the entire fat compartment contiguous with the ROOF. A two-tailed t test was conducted between ROOF and ROOF plus the extended fat plane across all measurements. A Wilcoxon nonparametric signed rank test was performed to determine equivalent fat distribution of the extended fat plane over each cadaver's respective eye. Results The deep fat originating from the ROOF consistently extended laterally and superiorly in each specimen, distinctly separated via septae from the deep central, deep lateral, and the deep temporal fat compartments. The color, composition, and distribution of this contiguous deep fat did not differ phenotypically from the traditional ROOF. The extended deep fat plane possessed an average vertical height of 3.09 ± 0.68 cm, average distance to midline from point of maximal height of 3.56 ± 0.53 cm, an average horizontal length of 5.37 ± 0.82 cm, and an average area of 13.40 ± 2.69 cm. The extended deep fat demonstrated a statistically significant increase in maximal height, length, and total area compared with the ROOF. A Wilcoxon nonparametric signed rank test was nonsignificant (α = 0.01) across all measurements, demonstrating that the extended fat plane was similarly distributed over each eye. Conclusions A layer of deep fat originating from the traditionally defined ROOF extends superiorly and laterally beneath the frontalis muscle, separate from the deep lateral, deep central, and deep temporal fat pads. This is the first study to clearly demonstrate a contiguous superficial musculoaponeurotic system layer of fat extending under both the orbicularis oculi and frontalis muscles. This plane of fat is more appropriately described as the retro-orbicularis oculi and frontalis fat.

Background Asthma was identified as the most common comorbidity in hospitalized patients during the 2009 H1N1 influenza pandemic. We determined using a murine model of allergic asthma whether these mice experienced increased morbidity from pandemic H1N1 (pH1N1) viral infection and whether blockade of interleukin-4 receptor α (IL-4Rα), a critical mediator of Th2 signalling, improved their outcomes. Methods Male BALB/c mice were intranasally sensitized with house dust mite antigen (Der p 1) for 2 weeks; the mice were then inoculated intranasally with a single dose of pandemic H1N1 (pH1N1). The mice were administered intraperitoneally anti-IL-4Rα through either a prophylactic or a therapeutic treatment strategy. Results Infection with pH1N1 of mice sensitized to house dust mite (HDM) led to a 24% loss in weight by day 7 of infection (versus 14% in non-sensitized mice; p Conclusion Together, these data implicate allergic asthma as a significant risk factor for H1N1-related morbidity and reveal a potential therapeutic role for IL-4Rα signalling blockade in reducing the severity of influenza infection in those with allergic airway disease.

Additional file 1: Table E1. Primers used for qPCR assays. Figure E1. Lung Tissue Sectioning. The left lung was cut in a transverse manner to create a superior and an inferior segment. The inferior segment was embedded into paraffin, while the superior segment was further cut in a sagittal manner and embedded. Figure E2. House dust mite (HDM) sensitization delays viral clearance. Phosphate-buffered saline (PBS)-exposed or HDM-sensitized mice were infected with pandemic H1N1 (pH1N1) and sacrificed on days 0, 2, 4, 6, and 8 post-infection. (A) IFN-β protein expression was determined using ELISA on the supernatant of bronchoalveolar lavage fluid (BALF). (B) qPCR was performed on RNA from homogenized lung tissue to determine the number of viral RNA copies, and results were compared using a two-tailed Student’s t-test. Data (n=5 per group per day) were compared using a two-way ANOVA with Sidak’s multiple comparison’s test and are expressed as mean ± SEM. *P

Growth of mammalian cells in the production of biotherapeutics often require the benefits of chemically defined media (CDM). Storage, handling and stability advantages of CDM powders govern the preponderance of their use across the industry. Physico-chemical property lot-to-lot variation of these multicomponent powders, however, continues to be a challenge. Process imposed degradation of amino acids and vitamins, for example, can influence cell density, specific titer, and the quality profile of the molecule expressed due to the lack of process understanding and suitable mitigation controls. Such degradation can materialize in either their manufacture or in downstream media dissolution steps. Colorimetry, in lieu of visual appearance, can be an effective surveillance method for the direct assessment of CDM quality as color change is indicative of chemical-physical variations. This work describes a series of studies aimed to establish relationships between quantitative color change and physico-chemical attribute variation of glucose-free and glucose-based powders. The results illustrate color change is indicative of amino acid glycation, vitamin degradation and particle size shifts. These relationships enable a colorimetric control strategy for the sensitive and rapid detection of relevant CDM variation to drive additional targeted assessments to improve the productivity and robustness of cell culture processes.

Analysis of fire debris for triglyceride-based oils may be of interest to fire investigators depending on the circumstances of a particular fire. Such circumstances include accidental or intentionally set cooking oil fires, fires involving triglyceride-based "eco" fire log products, and spontaneous ignition fires that involve drying oils on rags. Many forensic laboratories utilize gas chromatography-mass spectrometry following fatty acid methyl esterification to identify triglyceride residues in fire debris extracts. This study explores an alternate approach, which involves the identification of intact triglycerides by liquid chromatography-mass spectrometry-mass spectrometry (LC-MS/MS). 52 triglyceride-based oils and fats (22 different types) were analyzed by LC-MS/MS using multiple reaction monitoring to investigate variation in triglyceride content between different brands and types of oil and fat. Selected oils were then degraded by exposure to air, typical cooking conditions and/or fire conditions to simulate samples that are typically encountered by Fire Debris Analysts in fire investigation cases. Triglycerides were identified in all pristine and degraded oil samples, and relative peak areas for degraded samples often resembled their pristine oil counterparts. In samples where relative peak area differences were noted, more predominant degradation was observed for triglycerides with a higher proportion of poly-unsaturated fatty acids. Variability in triglyceride content between different brands and types of oil are discussed, as well as factors affecting the identification of triglyceride peaks in commercial oil samples, as compared to the corresponding analytical standard.

Chinese hamster ovary (CHO) cells are currently the primary host cell lines used in biotherapeutic manufacturing of monoclonal antibodies (mAbs) and other biopharmaceuticals. Cellular energy metabolism and endoplasmic reticulum (ER) stress are known to greatly impact cell growth, viability, and specific productivity of a biotherapeutic; but the molecular mechanisms are not fully understood. The authors previously employed multi-omics profiling to investigate the impact of a reduction in cysteine (Cys) feed concentration in a fed-batch process and found that disruption of the redox balance led to a substantial decline in cell viability and titer. Here, the multi-omics findings are expanded, and the impact redox imbalance has on ER stress, mitochondrial homeostasis, and lipid metabolism is explored. The reduced Cys feed activates the amino acid response (AAR), increases mitochondrial stress, and initiates gluconeogenesis. Multi-omics analysis reveals that together, ER stress and AAR signaling shift the cellular energy metabolism to rely primarily on anaplerotic reactions, consuming amino acids and producing lactate, to maintain energy generation. Furthermore, the pathways are demonstrated in which this shift in metabolism leads to a substantial decline in specific productivity and altered mAb glycosylation. Through this work, meaningful bioprocess markers and targets for genetic engineering are identified.

2648 Background: Despite an exponential increase in the number of potential targets in the immuno-oncology (IO) field, lack of risk models to predict toxicities remains a challenge in early drug development. We proposed a risk stratification strategy and investigated whether different IO classes may be associated with incremental risk of toxicities. Methods: A systematic search for IO studies from 01/2014 to 10/2020 was conducted. Among the 12053 abstracts screened, 254 reporting phase I-IO trials were selected. Type of IO treatment (IO monotherapy [mono] vs IO combination [comb]), therapeutic class (e.g. IO, molecularly targeted agents [MTA]), and dose escalation method (rule-based, model-based and model-assisted) were collected. A risk scoring model was developed after expert consensus: treatment-related deaths (1:yes, 0:no), incidence of G3/G4 treatment related adverse events (TRAE) or treatment emergent adverse events (TEAE) (0 if 1-29%,1 if ≥30-49%, 2 if ≥50%), incidence of ≥G2 cytokine release syndrome (1:yes, 0:no), incidence of ≥G2 encephalopathy (1:yes 0:no) and incidence of dose-limiting toxicity (DLT) (0:no, 1:lab, 2:clinical). Risk categories were defined by summing all points (0 = low, 1-2 = intermediate, 3+ = high) and were correlated with type of IO treatment, therapeutic class and dose escalation method. Results: Of 254 P1-IO trials reviewed, 228 (90%) were scorable, 26 were not (25 due to lack of AE data). Up to 10/26 (38%) of non-scorable studies were cell therapies. Among the 228 scorable studies, 120 (53%) scored 0, 65 (28%) scored 1-2, 43 (19%) scored 3+; 24 (11%) and 125 (55%) did not provide no. of pts with G3/G4 TRAEs or TEAEs respectively. A significant association was observed between risk categories and therapeutic class (p