Search

Culture-based microbial natural product discovery strategies fail to realize the extraordinary biosynthetic potential detected across earth's microbiomes. Here we introduce Small Molecule In situ Resin Capture (SMIRC), a culture-independent method to obtain natural products directly from the environments in which they are produced. We use SMIRC to capture numerous compounds including two new carbon skeletons that were characterized using NMR and contain structural features that are, to the best of our knowledge, unprecedented among natural products. Applications across diverse marine habitats reveal biome-specific metabolomic signatures and levels of chemical diversity in concordance with sequence-based predictions. Expanded deployments, in situ cultivation, and metagenomics facilitate compound discovery, enhance yields, and link compounds to candidate producing organisms, although microbial community complexity creates challenges for the later. This compound-first approach to natural product discovery provides access to poorly explored chemical space and has implications for drug discovery and the detection of chemically mediated biotic interactions. Environmental analyses predict extensive, yet to be realized natural product diversity. Herein, the authors report an approach that directly captures natural products from the environment, circumventing previous challenges and yielding compounds with unusual structures and activities. [ABSTRACT FROM AUTHOR]

Outcomes of PWH with unhealthy alcohol use, such as alcohol use reduction or progression to AUD, are not well-known and may differ by baseline patterns of unhealthy alcohol use. Among 1299 PWH screening positive for NIAAA-defined unhealthy alcohol use in Kaiser Permanente Northern California, 2013–2017, we compared 2-year probabilities of reduction to low-risk/no alcohol use and rates of new AUD diagnoses by baseline use patterns, categorized as exceeding: only daily limits (72% of included PWH), only weekly limits (17%), or both (11%), based on NIAAA recommendations. Overall, 73.2% (95% CI 70.5–75.9%) of re-screened PWH reduced to low-risk/no alcohol use over 2 years, and there were 3.1 (95% CI 2.5–3.8%) new AUD diagnoses per 100 person-years. Compared with PWH only exceeding daily limits at baseline, those only exceeding weekly limits and those exceeding both limits were less likely to reduce and likelier to be diagnosed with AUD during follow-up. PWH exceeding weekly drinking limits, with or without exceeding daily limits, may have a potential need for targeted interventions to address unhealthy alcohol use. [ABSTRACT FROM AUTHOR]

Philosophers of criminal punishment disagree about whether infliction of punishment for negligence can be morally justified. One contending view holds that it cannot be because punishment requires culpability and culpability requires, at a minimum, advertence to the facts that make one's conduct wrongful. Larry Alexander and Kim Ferzan are prominent champions of this position. This essay challenges that view and their arguments for it. Invoking a conceptual distinction between an agent's being blameworthy for an act and their deserving punishment (or suffering) for that act, it explains that an agent can be blameworthy for negligent conduct, and thus liable to reasonable blaming practices, even if negligence is not culpable, hence not sufficient to ground negative desert. Turning from conceptual inquiry to substantive questions of political morality, it then argues that a faulty actor's lack of culpability does not render them immune from just punishment, but does significantly limit the severity of punishment that may be inflicted, for punishment should not be disproportionately severe relative to an agent's culpability in relation to wrongdoing. [ABSTRACT FROM AUTHOR]

Neuronal ceroid lipofuscinoses (NCL; Batten disease) are a group of inherited neurodegenerative diseases with a common set of symptoms including cognitive and motor decline and vision loss. Naturally occurring sheep models of CLN5 and CLN6 disease display the key clinical features of NCL, including a progressive loss of vision. We assessed retinal histology, astrogliosis, and lysosomal storage accumulation in CLN5 affected (CLN5−/−) and CLN6 affected (CLN6−/−) sheep eyes and age-matched controls at 3, 6, 12, and 18 months of age to determine the onset and progression of retinal pathology in NCL sheep. The retina of CLN5−/− sheep shows progressive atrophy of the outer retinal layers, widespread gliosis, and accumulation of lysosomal storage in retinal ganglion cells late in disease. In contrast, CLN6−/− retina shows significant atrophy of all retinal layers, progressive gliosis, and earlier accumulation of lysosomal storage. This study has highlighted the differential vulnerability of retinal layers and the time course of retinal atrophy in two distinct models of NCL disease. This data will be valuable in determining potential targets for ocular therapies and the optimal timing of these therapies for protection from retinal dysfunction and degeneration in NCL. [ABSTRACT FROM AUTHOR]

Philosophers of criminal punishment widely agree that criminal punishment should be "proportional" to the "seriousness" of the offense. But this apparent consensus is only superficial, masking significant dissensus below the surface. Proposed proportionality principles differ on several distinct dimensions, including: (1) regarding which offense or offender properties determine offense "seriousness" and thus constitute a proportionality relatum; (2) regarding whether punishment is objectionably disproportionate only when excessively severe, or also when excessively lenient; and (3) regarding whether the principle can deliver absolute ("cardinal") judgments, or only comparative ("ordinal") ones. This essay proposes that these differences cannot be successfully adjudicated, and one candidate proportionality principle preferred over its rivals, in the abstract; a proportionality principle only makes sense as an integrated part of a more complete justificatory theory of criminal punishment. It then sketches a proportionality principle that best fits the responsibility-constrained pluralist theories of criminal punishment that currently predominate. The proportionality principle it favors provides that punishments should not be disproportionately severe, in noncomparative terms, relative to an agent's culpability in relation to their wrongdoing. [ABSTRACT FROM AUTHOR]

Age-related macular degeneration (AMD) is the primary cause of blindness in adults over 60 years of age, and clinical trials are currently assessing the therapeutic potential of retinal pigmented epithelial (RPE) cell monolayers on implantable scaffolds to treat this disease. However, challenges related to the culture, long-term storage, and long-distance transport of such implants currently limit the widespread use of adherent RPE cells as therapeutics. Here we report a xeno-free protocol to cryopreserve a confluent monolayer of clinical-grade, human embryonic stem cell-derived RPE cells on a parylene scaffold (REPS) that yields viable, polarized, and functional RPE cells post-thaw. Thawed cells exhibit ≥ 95% viability, have morphology, pigmentation, and gene expression characteristic of mature RPE cells, and secrete the neuroprotective protein, pigment epithelium-derived factor (PEDF). Stability under liquid nitrogen (LN2) storage has been confirmed through one year. REPS were administered immediately post-thaw into the subretinal space of a mammalian model, the Royal College of Surgeons (RCS)/nude rat. Implanted REPS were assessed at 30, 60, and 90 days post-implantation, and thawed cells demonstrate survival as an intact monolayer on the parylene scaffold. Furthermore, immunoreactivity for the maturation marker, RPE65, significantly increased over the post-implantation period in vivo, and cells demonstrated functional attributes similar to non-cryopreserved controls. The capacity to cryopreserve adherent cellular therapeutics permits extended storage and stable transport to surgical sites, enabling broad distribution for the treatment of prevalent diseases such as AMD. [ABSTRACT FROM AUTHOR]

Objective:To investigate whether sex differences exist in the pattern of change in C-reactive protein (CRP) levels during weight loss, and whether the associations between weight change and CRP change differ by the types of anthropometric variables.Design:Longitudinal, prospective analysis of subjects participating in an intentional weight loss trial (the Lose It For Ever: LIFE Study) followed-up for 30 months.Subjects:A total of 212 healthy, obese men and women (age: 23-77 years, body mass index (BMI): 30-39 kg m−2) took part in this study.Measurements:BMI, waist and hip circumferences, and waist-to-hip ratio, CRP and lifestyle variables repeatedly measured at baseline, 6, 12, 18 and 30-month follow-up.Results:Weight change was J shaped with a nadir at 12 months in both men and women (P for month2 <0.0001). CRP level was consistently higher in women than in men, but the differences were less prominent and were not statistically significant at 12- and 18-month follow-up. CRP changes between any two consecutive visits were significantly associated with changes in BMI during the same period in women. However, the associations between CRP changes and changes in waist or hip circumference were not as consistent, especially between 18- and 30-month follow-up when CRP significantly increased. The associations in men were generally similar among the different anthropometric measures. The association between changes in BMI and CRP was stronger in men than in women.Conclusion:BMI change generally correlated well with CRP changes in both men and women in the course of follow-up. Significant sex difference in CRP level at baseline diminished at 12- and 18-month follow-up, when both sexes had maintained the lost weight. [ABSTRACT FROM AUTHOR]

The article presents the author's comments on the arguments of Prof. Larry Alexander on the issue defense and justification. Prof. Alexander demonstrated that the necessity defense raises a host of the issues that have gone unnoticed and with respect to many, he aimed not merely to diagnose difficulties but also to prescribe the solutions. Prof. Alexander did not say merely that the necessity defense is an interesting defense, rather he justified a closer look at issues internal to the necessity defense by reference to the status of that defense within an overarching classification of criminal law defenses. The relationship between the status of necessity as a justificatory defense and the correct contours of the necessity defense, is not an explicit subject of this article. If the relationship is important, it is doing its work offstage. Prof. Alexander asserted that the mistake regarding lesser evils should deprive defendant of the status of justified and render him excused instead because necessity is a justificatory defense.

The stereological method of cross-counting based on the Cavalieri principle is widely used in neuroimaging to estimate the volume of cerebral structures. Although superficially simple, the stereological technique is validated by arcane mathematical proofs, so the cross size is determined by most investigators on a pragmatic basis with the assumption that the volume calculated is independent of the cross size used. We used three cross sizes (8×8, 5×5 and 3×3 voxels) to estimate the brain volume of six healthy control subjects. The volume estimate using a cross size of 3×3 was 9% larger than with a cross size of 5×5 and the latter was 15% larger than with a cross size of 8×8. We conclude that cross size significantly affects whole brain volume estimates and this result is presumably applicable to other structures whose cross-sectional areas form complex shapes. Investigators should be aware of this fact, especially when trying to make direct comparisons between volume estimates derived from the stereological method using different cross sizes. [ABSTRACT FROM AUTHOR]

Drug development is a risky business. Success or failure often depends on selecting one or two molecules for development from many choices offered by the engines of high-throughput discovery. A lead candidate needs to possess adequate bioactivity, appropriate physical-chemical properties to enable formulation development, the ability to cross crucial membranes, reasonable metabolic stability and appropriate safety and efficacy in humans. Predicting how a drug will behave in humans before clinical testing requires a battery of sophisticated in vitro tests that complement traditional in vivo animal safety assessments. This review discusses how to strategically identify which non-clinical studies should be performed to provide the required guidance and comfort to stakeholders involved in clinical drug testing. [ABSTRACT FROM AUTHOR]

The ITER magnet system consists of structurally linked sets of toroidal (TF) and poloidal (PF) field coils, central solenoid (CS), and various support structures. The coils are superconducting, force flow Helium cooled with a Kapton-Glass-Epoxy multilayer insulation system. The stored magnetic energy is about 100GJ in the TF system and 20GJ in the PF-CS. Coils and structure are maintained at 4 K by enclosing them in a vacuum cryostat. The cryostat, comprising an outer envelope to the magnets, forms most of the second radioactivity confinement barrier. The inner primary barrier is formed by the vacuum vessel, its ports and their extensions. To keep the machine size within acceptable bounds, it is essential that the magnets are in close proximity to both of the nuclear confinement barriers. The objective of the magnet design is that, although local damage to one of the barriers may occur in very exceptional circumstances, large scale magnet structural or thermal failure leading to simultaneous breaching of both barriers is not credible. Magnet accidents fall into three categories: thermal (which includes arcing arising from insulation failure and local overheating due to discharge failure in the event of a superconductor quench), structural (which includes component mechanical failure arising from material inadequacies, design errors and exceptional force patterns arising from coil shorts or control failures), and fluid (Helium release due to cooling line failure). After a preliminary survey to select initial faults conceivable within the present design, these faults are systematically analyzed to provide an assessment of the damage potential. The results of this damage assessment together with an assessment of the reliability of the monitoring and protective systems, shows that the magnets can operate with the required safety condition. [ABSTRACT FROM AUTHOR]

A newly diagnosed patient with chronic lymphocytic leukemia who presented with ascites is described. Paracentesis revealed the presence of massive chylous ascites. Further work-up disclosed leukemic infiltration of the liver and bone marrow. Complications leading to his death included renal failure, gastrointestinal hemorrhage, gastrointestinal hemorrhitis, and Staphylococcus aureus bacteremia. Chylous ascites is a rarely reported manifestation of chronic lymphocytic leukemia. The case is presented, and a review of the literature is discussed. [ABSTRACT FROM AUTHOR]

The wavelength dependence of the fragmentation of gallstones was investigated using pulsed excimer, ruby and Nd-YAG lasers. Fracturing is due to the shock waves produced by the plasma formed at the irradiated surface. The fluence threshold for plasma production and the associated acoustic signal was found to depend on the absorption coefficient at the laser wavelength. Using a simple model based on a thermal mechanism for plasma production the fluence threshold at different wavelengths can be predicted and the effect of the duration of the irradiating pulse can be qualitatively explained. [ABSTRACT FROM AUTHOR]

This study reports a method whereby glycogen is identified in the chondrocytes of the secondary center of ossification prior to mineralization. The use of new fuchsin rather than basic fuchsin on one micron Spurr sections of femoral head cartilage fixed with potassium ferrocyanide-reduced osmium produced excellent identification of glycogen and when followed by p phenylenediamine, intensified cellular detail. [ABSTRACT FROM AUTHOR]

The sulphur content as a measure of glycosaminoglycan content of growth plate cartilage was determined by energy dispersive x-ray analysis on fresh freeze dried unstained, unfixed ultra thin sections of rat growth plate. In the resting and proliferative zones, quantities of sulphur were found in the nuclei equal to that of the matrix. Less sulphur was present in the cytoplasm. In areas of cell degeneration, nuclear and cytoplasmic content of sulphur fell to levels a fraction of that seen in the matrix. It was presumed that most of the sulphur was in glycosaminoglycans. Although glycosaminoglycans have been reported in small amounts in the nuclei of cells, no study of the glycosaminoglycan content of chondrocyte nuclei has been reported. The use of freeze dried unstained, unfixed sections presumably prevented the migration of sulphur and glycosaminoglycans from compartment to compartment. [ABSTRACT FROM AUTHOR]

This study indicated that acridine orange, when combined with the initial fixative stabilized soluble matrix glycosaminoglycan in situ in areas where considerable glycosaminoglycan extraction is known to occur. Acridine orange was able to diffuse through bone into areas of undecalcified mineralizing cartilage and to bind with the glycosaminoglycans in these areas equally well as in growth plate cartilage matrix. Matrix staining was visible by light microscopy without further staining and was seen to vary territorially in intensity; although cellular definition was poor. This deficiency was overcome by the additional application of p phenylenediamine, which stained the cells intensely. At the ultrastructure level, glycosaminoglycan was present as electron dense structures in the cartilage matrix. Preliminary X-ray microanalysis studies confirmed that the acridine orange stained structures contain sulphur; this finding extends the use of acridine orange further to quantitative analysis of glycosaminoglycan. [ABSTRACT FROM AUTHOR]

Toluidine blue O, a cationic dye has been used for visualizing proteoglycan by light and electron microscopy. Toluidine blue O has now been brominated and incorporated into epiphyseal cartilage during its fixation. Using energy dispersive X-ray microanalysis, bromine has been measured from area to area and compared to measurements of sulphur, a naturally occurring element of proteoglycan. Since bromine has been combined mole for mole with toluidine blue O and it is already known that toluidine blue O combines approximately mole for mole with the glycosaminoglycans of proteoglycan, the measurement of bromine, presents an indirect measure of proteoglycan concentration. The X-ray spectral localization of sulphur presents problems in X-ray analysis which are overcome with bromine which makes this technique attractive for workers in this field. [ABSTRACT FROM AUTHOR]

A 2°×2° map of spreading centres and fracture zones surrounding the Indian Ocean RRR triple junction, at 25.5°S, 70°E, is described from a data set of GLORIA side-scan sonar images, bathymetry, magnetic and gravity anomalies. The GLORIA images show a pervasive fabric due to linear abyssal hills oriented parallel to the two medium-spreading ridges (the Central Indian Ridge (CIR) and Southeast Indian Ridge (SEIR)). A cuvature of the fabric occurs along fracture zones, which are also located by lows in the bathymetry and gravity data and by offsets between magnetic anomalies. The magnetic anomalies also record periods of asymmetric spreading marking the development of the fracture zones, including the birth, at anomaly 2A, of a short fracture zone 50 km north of the triple junction on the CIR, and its death near the time of the Jaramillo anomaly. In some localities, a fine-scale fabric corresponds to a coarser fabric on the opposite flank of the CIR, possibly indicating a persistent asymmetry in the faulting at the median valley walls if the fabric has a tectonic and not a volcanic origin. A plate velocity analysis of the triple junction shows that both the CIR and Southwest Indian Ridge (SWIR) are propagating obliquely; the CIR appears to form an oblique trend by segmenting into a series of almost normally-oriented segments separated by short-offset fracture zones. For the last 4 m.y., the abyssal hill lineations indicate that the CIR segment immediately north of the triple junction has been spreading with an average 10° obliquity. The present small 5 km offset of the centres of the CIR and SEIR median valleys (Munschy and Schlich, 1989) is shown to be the result of this obliquity and a 30% spreading asymmetry between anomaly 2 and the Jaramillo on the CIR segment immediately north of the triple junction. [ABSTRACT FROM AUTHOR]

This chapter presents a summary of the image-processing techniques being used at present in the Institute of Oceanographic Sciences Deacon Laboratory's GLORIA long-range sidescan sonar system. It begins with a brief review of the development of GLORIA, and then describes in outline the present shipboard data acquisition, recording and replay system, including simple image-processing techniques that can be used on-board ship. Next, a detailed form of the sonar equation is developed, and this is evaluated factor-by-factor, to demonstrate the effects of beam directivity, refraction and water depth on the form of intensity variation to be expected in the final image. Finally, we discuss recent developments in shore-based image-processing. These include the development of improved radiometric corrections to normalize range-dependent intensity variations, recovery of true backscattering levels and estimation of backscattering coefficients, and combination of GLORIA with other data sets into single, colour digital images. As an example of the last process we show a digital mosaic of sonar data from the Southwest Indian Ridge, coloured as a function of depth derived from Sea Beam data in the same area. [ABSTRACT FROM AUTHOR]

Massive osteolysis is a rare condition and is very uncommon in the spine. The MRI appearance of Gorham's disease of the spine has not previously been reported. We present here a case of this condition with imaging details. [ABSTRACT FROM AUTHOR]