Recessive variants in the intergenic NOS1AP-C1orf226 locus cause monogenic kidney disease responsive to anti-proteinuric treatment.

In genetic disease, an accurate expression landscape of disease genes and faithful animal models will enable precise genetic diagnoses and therapeutic discoveries, respectively. We previously discovered that variants in NOS1AP , encoding nitric oxide synthase 1 (NOS1) adaptor protein, cause monogenic nephrotic syndrome (NS). Here, we determined that an intergenic splice product of N OS1AP / Nos1ap and neighboring C1orf226/Gm7694 , which precludes NOS1 binding, is the predominant isoform in mammalian kidney transcriptional and proteomic data. Gm7694 -/- mice, whose allele exclusively disrupts the intergenic product, developed NS phenotypes. In two human NS subjects, we identified causative NOS1AP splice variants, including one predicted to abrogate intergenic splicing but initially misclassified as benign based on the canonical transcript. Finally, by modifying genetic background, we generated a faithful mouse model of NOS1AP -associated NS, which responded to anti-proteinuric treatment. This study highlights the importance of intergenic splicing and a potential treatment avenue in a mendelian disorder.

Feeding frequency does not interact with BPA exposure to influence metabolism or behaviour in zebrafish (Danio rerio).

Resource limitation can constrain energy (ATP) production, and thereby affect locomotion and behaviour such as exploration of novel environments and boldness. Consequently, ecological processes such as dispersal and interactions within and between species may be influenced by food availability. Energy metabolism, and behaviour are regulated by endocrine signalling, and may therefore be impacted by endocrine disrupting compounds (EDCs) including bisphenol A (BPA) derived from plastic manufacture and pollution. It is important to determine the impacts of these novel environmental contexts to understand how human activity alters individual physiology and behaviour and thereby populations. Our aim was to determine whether BPA exposure interacts with feeding frequency to alter metabolism and behaviour. In a fully factorial experiment, we show that low feeding frequency reduced zebrafish (Danio rerio) mass, condition, resting metabolic rates, total distance moved and speed in a novel arena, as well as anxiety indicated by the number of times fish returned to a dark shelter. However, feeding frequency did not significantly affect maximal metabolic rates, aerobic scope, swimming performance, latency to leave a shelter, or metabolic enzyme activities (citrate synthase and lactate dehydrogenase). Natural or anthropogenic fluctuation in food resources can therefore impact energetics and movement of animals with repercussions for ecological processes such as dispersal. BPA exposure reduced LDH activity and body mass, but did not interact with feeding frequency. Hence, behaviour of adult fish is relatively insensitive to disruption by BPA. However, alteration of LDH activity by BPA could disrupt lactate metabolism and signalling and together with reduction in body mass could affect size-dependent reproductive output. BPA released by plastic manufacture and pollution can thereby impact conservation and management of natural resources.

Bisphenol S reduces locomotor performance and modifies muscle protein levels but not mitochondrial bioenergetics in adult zebrafish.

Human activity has now introduced novel chemicals into most aquatic ecosystems. Endocrine-disrupting compounds originating from plastic pollution and manufacture can have pronounced biological effects by disrupting hormone-mediated processes. Bisphenol A (BPA) is one of the most commonly produced endocrine-disrupting compounds, which interferes with signalling by a broad range of hormones. In recognition of its potentially harmful effects, BPA is being replaced by substitutes such as bisphenol S (BPS). However, toxicological studies revealed that BPS too can bind to hormone receptors and disrupt signalling, particularly of thyroid hormone. The aim of this study was to test whether BPS exposure impacts locomotor performance and muscle function in zebrafish (Danio rerio). Locomotor performance depends on thyroid hormone signalling, and it is closely related to fitness so that its disruption can have negative ecological and evolutionary consequences. BPS exposure of 15 µg l-1 [∼60 nM] and 30 µg l-1 (but not 60 µg l-1) decreased sustained swimming performance (Ucrit), but not sprint speed. In a fully factorial design, we show that living in flowing water increased Ucrit compared to a still water control, and that BPS reduced Ucrit under both conditions but did not eliminate the training effect. In a second factorial experiment, we show that BPS did not affect mitochondrial bioenergetics in skeletal muscle (state 3 and 4 rates, respiratory control ratios, ROS production), but that induced hypothyroidism decreased state 3 and 4 rates of respiration. However, both hypothyroidism and BPS exposure decreased activity of AMP-activated protein kinase (pAMPK:total AMPK) but increased protein levels of myocyte enhancer factor 2, and slow and fast myosin heavy chains. Our data indicate that BPS is not a safe alternative for BPA and that exposure to BPS can have ecological consequences, which are likely to be at least partly mediated via thyroid hormone disruption.

Bisphenols impact hormone levels in animals: A meta-analysis.

Bisphenols are used in the manufacture of plastics and are endocrine disrupting compounds detectable in free living organisms and environments globally. The original bisphenol, bisphenol A (BPA), is best known as a xenoestrogen, but it also disrupts other steroid hormones and other classes of hormones including thyroid and pituitary hormones. When its toxicity became better known, BPA was replaced by presumably less toxic alternatives, including bisphenols S, F, and AF. However, recent data suggest that all bisphenols can have endocrine disrupting effects, although their impacts remain unresolved particularly in non-human animals. Our aim was to establish the current state-of-knowledge of the effects of different bisphenols on circulating hormone levels in non-human animals. Our meta-analysis showed that a diverse range of hormones (including thyroid hormones, corticosterone, follicle stimulating hormone, luteinizing hormone, and estradiol) are strongly impacted by exposure to any bisphenol type, and that in laboratory rats (Rattus norvegicus) the effect was modified by life-stage. Although there were qualitative differences, BPA alternatives had as great or greater effects on hormone levels as BPA. However, data coverage across hormones was uneven, and most studies measured the effects of BPA on vertebrate reproductive hormones. Similarly, taxonomic coverage was poor. Over 80% of data originated from laboratory rats and zebrafish (Danio rerio) and there are no data for whole classes of invertebrates and vertebrates (e.g., amphibians). Our results show that all bisphenols alter circulating levels of a broad range of hormones. However, the current state-of-knowledge is incomplete so that the ecological impacts of bisphenols are difficult to gauge, although based on the available data bisphenols are likely to be detrimental to a broad range of taxa and ecosystems.

Postnatal expression of IGF2 is the norm in amniote vertebrates.

The insulin and insulin-like signalling (IIS) network plays an important role in mediating several life-history traits, including growth, reproduction and senescence. Although insulin-like growth factors (IGFs) 1 and 2 are both key hormones in the vertebrate IIS network, research on IGF2 in juveniles and adults has been largely neglected because early biomedical research on rodents found negligible IGF2 postnatal expression. Here, we challenge this assumption and ask to what degree IGF2 is expressed during postnatal life across amniotes by quantifying the relative gene expression of IGF1 and IGF2 using publicly available RNAseq data for 82 amniote species and quantitative polymerase chain reaction on liver cDNA at embryonic, juvenile and adult stages for two lizard, bird and mouse species. We found that (i) IGF2 is expressed postnatally across amniote species and life stages-often at a higher relative expression than IGF1, contradicting rodent models; (ii) the lack of rodent postnatal IGF2 expression is due to phylogenetic placement, not inbreeding or artificial selection; and (iii) adult IGF2 expression is sex-biased in some species. Our results demonstrate that IGF2 expression is typical for amniotes throughout life, suggesting that a comprehensive understanding of the mechanisms mediating variation in life-history traits will require studies that measure both IGFs.

Endocrine disruption from plastic pollution and warming interact to increase the energetic cost of growth in a fish.

Energetic cost of growth determines how much food-derived energy is needed to produce a given amount of new biomass and thereby influences energy transduction between trophic levels. Growth and development are regulated by hormones and are therefore sensitive to changes in temperature and environmental endocrine disruption. Here, we show that the endocrine disruptor bisphenol A (BPA) at an environmentally relevant concentration (10 µgl-1) decreased fish (Danio rerio) size at 30°C water temperature. Under the same conditions, it significantly increased metabolic rates and the energetic cost of growth across development. By contrast, BPA decreased the cost of growth at cooler temperatures (24°C). BPA-mediated changes in cost of growth were not associated with mitochondrial efficiency (P/O ratios (i.e. adenosine diphosphate (ADP) used/oxygen consumed) and respiratory control ratios) although BPA did increase mitochondrial proton leak. In females, BPA decreased age at maturity at 24°C but increased it at 30°C, and it decreased the gonadosomatic index suggesting reduced investment into reproduction. Our data reveal a potentially serious emerging problem: increasing water temperatures resulting from climate warming together with endocrine disruption from plastic pollution can impact animal growth efficiency, and hence the dynamics and resilience of animal populations and the services these provide.

Genetic contribution to high temperature tolerance in Cryptococcus neoformans.

The human fungal pathogen Cryptococcus neoformans relies on a complex signaling network for the adaptation and survival at the host temperature. Protein phosphatase calcineurin is central to proliferation at 37°C but its exact contributions remain ill-defined. To better define genetic contributions to the C. neoformans temperature tolerance, 4031 gene knockouts were screened for genes essential at 37°C and under conditions that keep calcineurin inactive. Identified 83 candidate strains, potentially sensitive to 37°C, were subsequently subject to technologically simple yet robust assay, in which cells are exposed to a temperature gradient. This has resulted in identification of 46 genes contributing to the maximum temperature at which C. neoformans can proliferate (Tmax). The 46 mutants, characterized by a range of Tmax on drug-free media, were further assessed for Tmax under conditions that inhibit calcineurin, which led to identification of several previously uncharacterized knockouts exhibiting synthetic interaction with the inhibition of calcineurin. A mutant that lacked septin Cdc11 was among those with the lowest Tmax and failed to proliferate in the absence of calcineurin activity. To further define connections with calcineurin and the role for septins in high temperature growth, the 46 mutants were tested for cell morphology at 37°C and growth in the presence of agents disrupting cell wall and cell membrane. Mutants sensitive to calcineurin inhibition were tested for synthetic lethal interaction with deletion of the septin-encoding CDC12 and the localization of the septin Cdc3-mCherry. The analysis described here pointed to previously uncharacterized genes that were missed in standard growth assays indicating that the temperature gradient assay is a valuable complementary tool for elucidating the genetic basis of temperature range at which microorganisms proliferate.

Periodic Cooling during Incubation Alters the Adrenocortical Response and Posthatch Growth in Zebra Finches.

AbstractIn birds, incubation temperature is critically deterministic for a range of traits. When parents leave the nest to forage, developing embryos can be exposed to cooling events that represent thermal stress. To investigate the consequences of periodic cooling on offspring development and physiology, we exposed zebra finch embryos to cooling events throughout the incubation period. We then compared embryonic survival, egg mass change, incubation duration, posthatch growth, and adrenocortical response of these individuals with embryos reared at a constant optimal temperature of 37.4°C and embryos reared at a constant suboptimal temperature of 36.4°C, the mean incubation temperature of periodically cooled embryos. There were no differences in embryonic survival or egg mass change during incubation, but individuals exposed to periodic cooling had longer incubation periods than those from the 37.4°C treatment and shorter incubation periods than those from the 36.4°C treatment. Periodically cooled individuals showed slower posthatch growth in comparison with both constant-temperature treatments, but this did not impact adult body size. Treatment groups did not differ in their adrenocortical response, but embryos exposed to periodic cooling and a constant temperature of 37.4°C were able to habituate to repeated capture and restraint stress, while individuals exposed to the constant temperature of 36.4°C were not. These results point to the differential impacts of cooling events versus constant low temperatures during incubation on posthatch growth and physiology and may represent a way for parents to devote less energy toward incubation while still ensuring offspring success.

Nonbacterial thrombotic endocarditis in a patient with primary antiphospholipid syndrome.

Nonbacterial thrombotic endocarditis (NBTE) is described in patients with mucin-producing cancers and connective tissue disorders (usually SLE). We report NBTE in the setting of primary antiphospholipid antibody syndrome (APS). A 65-year-old female with APS was incidentally found to have thickened mitral leaflets on transthoracic echocardiogram with no signs of infection. Transesophageal echocardiogram (TEE) showed a mobile mitral mass (1.4 × 0.7 cm) and moderate mitral regurgitation. Differential diagnoses included bacterial endocarditis, NBTE, thrombus or tumor. Given the history of primary APS, the absence of fever and negative blood cultures, NBTE was considered. Low-molecular-weight heparin, hydroxychloroquine and corticosteroid were initiated. Repeat TEE in a week revealed shrinkage of the mass (0.6 × 0.7 cm), indicating an inflammatory nature. Lifelong anticoagulation is indicated regardless of embolism occurrence. Hydroxychloroquine and corticosteroids may have roles in the treatment. Determining and treating the underlying etiology is important.

Endocarditis of the native aortic valve caused by Lactobacillus jensenii.

Lactobacilli are Gram-positive anaerobic rods or coccobacilli, commonly found as commensals in human mucosa. Rarely, they can cause serious infections such as infective endocarditis (IE), and the most frequently implicated species causing serious infections are L. casei and L. rhamnosus. IE caused by Lactobacillus jensenii is very rare, with only six reported cases so far, to the best of our knowledge. We present a case of native aortic valve endocarditis caused by L. jensenii, complicated by root abscess and complete heart block, and requiring emergent surgical intervention.

Thrombotic thrombocytopenic purpura and cardiac papillary fibroelastoma: a 'unique coexistence'.

Thrombotic thrombocytopenic purpura (TTP), a complex thrombotic microangiopathy, remains an evolving enigma. A 49-year-old African-American woman presented with acute left hemiplegia, an ischemic cerebrovascular accident involving the right middle cerebral artery. Sequential appearance of thrombocytopenia and evidence of microangiopathic haemolysis led to the diagnosis of acquired idiopathic autoimmune TTP. This was managed with plasma exchange (PEX) and steroids. Early haematologic relapse within a month was managed with the addition of rituximab attaining sustained remission. The patient presented 3 years later with acute confusion and expressive aphasia due to multiple infarcts involving the left parieto-occipital cortex. Transoesophageal echocardiography demonstrated a pedunculated 6 mm mitral valvular mass consistent with a papillary fibroelastoma. Anticoagulation was instituted and the patient was continued on therapeutic oral warfarin. A haematologic relapse of TTP eventually emerged and was managed with PEX, steroids and rituximab. This vignette demonstrates several dilemmas in the clinical presentation, diagnosis and management of TTP in current day practice. Rituximab has adjuvant benefits to PEX and is being investigated as potential first-line therapy. Monitoring ADAMTS13 activity and inhibitor titre, as in our case, prove to have prognostic significance. Cardiac fibroelastomas are rare benign cardiac tumours usually arising from valvular endocardium with thromboembolic potential. One of the proposed mechanisms of origin of these masses is organizing thrombi in the setting of endocardial injury and inflammation questioning a possible link to thrombotic microangiopathy. To the best of our knowledge, this is the first report of this unique coexistence.

Relationship of inferior vena cava filter usage in post-surgical patients by various surgical and medical subspecialists.

Venous thromboembolism is a common and often fatal problem in postsurgical patients. These patients are usually treated with either therapeutic anticoagulation or the placement of inferior vena cava (IVC) filters. Controversy surrounds the use of IVC filters, because no data exist proving survival benefit. In this study, 264 inpatient medical records of patients who underwent major surgical procedures and had the diagnosis of deep venous thrombosis or pulmonary embolism were examined. Among these patients, those who received IVC filters were identified, and the documented indications for filter placement were reviewed. Rates of IVC filter placement per venous thromboembolism event and specific indications were examined across surgical subspecialties and by type of medical consultant. Sixty percent of patients received IVC filters. IVC filter placement rates varied significantly across surgical subspecialties (p <0.0001), with the highest rate in the orthopedic surgery subgroup (80%). Rates of IVC filter use also differed significantly (p <0.0007) between medical consultants who specialized in antithrombotic medicine (46.8%) and those who did not (68.3%). Significant differences also existed in specific indications for filter placement between medical and surgical subspecialties. In conclusion, most of this study's population received IVC filters. Rates of IVC filter placement varied by the specialties of surgeons and medical consultants. The heterogeneity of treatment strategies coupled with the lack of data for this patient population highlights the need for future prospective studies to guide evidence-based treatment.