Adaptation of a transmitted/founder simian-human immunodeficiency virus for enhanced replication in rhesus macaques.

Transmitted/founder (TF) simian-human immunodeficiency viruses (SHIVs) express HIV-1 envelopes modified at position 375 to efficiently infect rhesus macaques while preserving authentic HIV-1 Env biology. SHIV.C.CH505 is an extensively characterized virus encoding the TF HIV-1 Env CH505 mutated at position 375 shown to recapitulate key features of HIV-1 immunobiology, including CCR5-tropism, a tier 2 neutralization profile, reproducible early viral kinetics, and authentic immune responses. SHIV.C.CH505 is used frequently in nonhuman primate studies of HIV, but viral loads after months of infection are variable and typically lower than those in people living with HIV. We hypothesized that additional mutations besides Δ375 might further enhance virus fitness without compromising essential components of CH505 Env biology. From sequence analysis of SHIV.C.CH505-infected macaques across multiple experiments, we identified a signature of envelope mutations associated with higher viremia. We then used short-term in vivo mutational selection and competition to identify a minimally adapted SHIV.C.CH505 with just five amino acid changes that substantially improve virus replication fitness in macaques. Next, we validated the performance of the adapted SHIV in vitro and in vivo and identified the mechanistic contributions of selected mutations. In vitro, the adapted SHIV shows improved virus entry, enhanced replication on primary rhesus cells, and preserved neutralization profiles. In vivo, the minimally adapted virus rapidly outcompetes the parental SHIV with an estimated growth advantage of 0.14 days-1 and persists through suppressive antiretroviral therapy to rebound at treatment interruption. Here, we report the successful generation of a well-characterized, minimally adapted virus, termed SHIV.C.CH505.v2, with enhanced replication fitness and preserved native Env properties that can serve as a new reagent for NHP studies of HIV-1 transmission, pathogenesis, and cure.

Measurement of Penrose Superradiance in a Photon Superfluid.

The superradiant amplification in the scattering from a rotating medium was first elucidated by Sir Roger Penrose over 50 years ago as a means by which particles could gain energy from rotating black holes. Despite this fundamental process being ubiquitous also in wave physics, it has only been observed once experimentally, in a water tank. Here, we measure this amplification for a nonlinear optics experiment in the superfluid regime. In particular, by focusing a weak optical beam carrying orbital angular momentum onto the core of a strong pump vortex beam, negative norm modes are generated and trapped inside the vortex core, allowing for amplification of a reflected beam. Our experiment demonstrates amplified reflection due to a novel form of nonlinear optical four-wave mixing, whose phase-relation coincides with the Zel'dovich-Misner condition for Penrose superradiance in our photon superfluid, and unveil the role played by negative frequency modes in the process.

Collective excitability in highly diluted random networks of oscillators.

We report on collective excitable events in a highly diluted random network of non-excitable nodes. Excitability arises thanks to a self-sustained local adaptation mechanism that drives the system on a slow timescale across a hysteretic phase transition involving states with different degrees of synchronization. These phenomena have been investigated for the Kuramoto model with bimodal distribution of the natural frequencies and for the Kuramoto model with inertia and a unimodal frequency distribution. We consider global and partial stimulation protocols and characterize the system response for different levels of dilution. We compare the results with those obtained in the fully coupled case showing that such collective phenomena are remarkably robust against network diluteness.

Development of a new, simple, rapid ultra-high-performance liquid chromatography (UHPLC) method for the quantification of 2-phenoxyethanol in vaccines for human use.

A new, simple and rapid method for the quantitative determination of the antimicrobial preservative 2-phenoxyethanol, based on reverse phase ultra-high-performance liquid chromatography has been developed. The validation was performed according the ICH Q2 guideline "Validation of Analytical Procedures". The desired chromatographic separation was achieved on a Waters Symmetry C18 (150 × 4.6 mm, 5 µm) column using an isocratic elution, with detection at 270 nm wavelength. The mobile phase consisted of acetonitrile/water (55:45, v/v), pumped at a flow rate of 1 mL/min. The calibration curve and the analytical procedure are linear (r2 = 0.999) from the concentration of 0.07 mg/mL to 1.1 mg/mL. The percent relative standard deviation for intra- and inter-day precision was <1%. The recovery of 2-phenoxyethanol in vaccines ranged between 96.5 and 100.60%. The limits of detection and quantitation were 1.3 × 10-4 and 2.7 × 10-4 mg/mL, respectively. The method was found to be robust by changing the column working temperature, the percentage of acetonitrile of the mobile phase and the flow rate. The validated method can be successfully and reliably used to quantify as well as to exclude presence of 2-phenoxyethanol preservative in marketed vaccines.

Association between antithrombin and mortality in patients with COVID-19. A possible link with obesity.

BACKGROUND AND AIMS: Despite anticoagulation, usually with heparin, mortality for thromboembolic events in COVID-19 remains high. Clinical efficacy of heparin is due to its interaction with antithrombin (AT) that may be decreased in COVID-19. Therefore, we correlated AT levels with outcomes of COVID-19. METHODS AND RESULTS: We recruited 49 consecutive patients hospitalized for COVID-19. AT levels were significantly lower in 16 non-survivors than in 33 survivors (72.2 ± 23.4 versus 94.6 ± 19.5%; p = 0.0010). A multivariate Cox regression analysis showed that low AT (levels below 80%) was a predictor of mortality (HR:3.97; 95%CI:1.38 to 11.43; p = 0.0103). BMI was the only variable that showed a significant difference between patients with low and those with normal AT levels (32.9 ± 7.9 versus 27.5 ± 5.9%; p = 0.0104). AT levels were significantly lower in obese patients than in subjects with normal weight or overweight (77.9 ± 26.9 versus 91.4 ± 26.9 versus 91.4 ± 17.1%; p = 0.025). An inverse correlation between AT levels and BMI was documented (r:-0.33; p = 0.0179). CONCLUSIONS: Our data first suggest that AT is strongly associated with mortality in COVID-19. In addition, AT may be the link between obesity and a poorer prognosis in patients with COVID-19. Other studies should confirm whether AT may become a prognostic marker and a therapeutic target in COVID-19.

Excitable Wave Patterns in Temporal Systems with Two Long Delays and their Observation in a Semiconductor Laser Experiment.

Excitable waves arise in many spatially extended systems of either a biological, chemical, or physical nature due to the interplay between local reaction and diffusion processes. Here we demonstrate that similar phenomena are encoded in the time dynamics of an excitable system with two, hierarchically long delays. The transition from 1D localized structures to curved wave segments is experimentally observed in an excitable semiconductor laser with two feedback loops and reproduced by numerical simulations of a prototypical model. While closely related to those found in 2D excitable media, wave patterns in delayed systems exhibit unobserved features originating from causality related constraints. An appropriate dynamical representation of the data uncovers these phenomena and permits us to interpret them as the result of an effective 2D advection-reaction-diffusion process.

Evidence of a Critical Phase Transition in Purely Temporal Dynamics with Long-Delayed Feedback.

Experimental evidence of an absorbing phase transition, so far associated with spatiotemporal dynamics, is provided in a purely temporal optical system. A bistable semiconductor laser, with long-delayed optoelectronic feedback and multiplicative noise, shows the peculiar features of a critical phenomenon belonging to the directed percolation universality class. The numerical study of a simple, effective model provides accurate estimates of the transition critical exponents, in agreement with both theory and our experiment. This result pushes forward a hard equivalence of nontrivial stochastic, long-delayed systems with spatiotemporal ones and opens a new avenue for studying out-of-equilibrium universality classes in purely temporal dynamics.

Role of activin C in normal ovaries and granulosa cell tumours of mice and humans.

Activins and inhibins play important roles in the development, growth and function of the ovary. Mice lacking inhibin develop granulosa cell tumours in their ovaries that secrete activin A, and these tumours are modulated by increased activin C expression. The aim of the present study was to identify where activin C is expressed in mouse and human ovaries and whether overexpression of activin C modulates normal follicular development in mice. Immunohistochemical staining for the activin ßC subunit was performed on sections from mouse and human ovaries and human adult granulosa cell tumours. Stereology techniques were used to quantify oocyte and follicular diameters, and the percentage of different follicular types in ovaries from wild-type mice and those underexpressing inhibin α and/or overexpressing activin C. Staining for activin ßC was observed in the oocytes, granulosa cells, thecal cells and surface epithelium of mouse and human ovaries, and in the granulosa-like cells of adult granulosa cell tumours. Overexpression of activin C in mice did not alter follicular development compared with wild-type mice, but it did modulate the development of abnormal early stage follicles in inhibin α-null mice. These results provide further evidence of a role for activin C in the ovary.

Sedative effects of intramuscular alfaxalone in pet guinea pigs (Cavia porcellus).

OBJECTIVE: To evaluate the efficacy and side effects of alfaxalone administered intramuscularly (IM) as a sedative agent in guinea pigs undergoing survey radiographs. STUDY DESIGN: Prospective clinical trial. ANIMALS: A total of 30 client-owned guinea pigs. METHODS: Following baseline assessments, 5 mg kg-1 alfaxalone was administered IM. Heart rate, arterial haemoglobin oxygen saturation, respiratory rate, rectal body temperature, palpebral reflex, response to toe and ear pinch, righting reflex, posture, jaw tone and reaction to manipulation were assessed before and after sedation at 5-minute intervals. The time elapsed from onset of sedation to return of locomotion and coordinated limb movements, the quality of recovery and the occurrence of undesired effects were observed and recorded. RESULTS: The mean ± standard deviation onset of sedation was 2.7 ± 0.6 minutes. The physiological variables remained within normal ranges until completion of the procedure. Palpebral reflex and responsiveness to both ear and toe pinch were maintained during sedation. Neither hypoxaemia nor hypothermia was observed. The duration of sedation was 29.3 ± 3.2 minutes. Sedation and recovery were uneventful, and adverse effects were not observed. CONCLUSIONS AND CLINICAL RELEVANCE: In conclusion, 5 mg kg-1 of IM alfaxalone represents a valuable sedation protocol for healthy guinea pigs undergoing minor noninvasive procedures. Further trials are required to investigate its cardiovascular effects, clinical usefulness in unhealthy patients and its combined use with analgesics for procedures associated with nociception.

Exogenous GDF11 induces cardiac and skeletal muscle dysfunction and wasting.

Growth differentiation factor 11 (GDF11), a TGF-beta superfamily member, is highly homologous to myostatin and essential for embryonic patterning and organogenesis. Reports of GDF11 effects on adult tissues are conflicting, with some describing anti-aging and pro-regenerative activities on the heart and skeletal muscle while others opposite or no effects. Herein, we sought to determine the in vivo cardiac and skeletal muscle effects of excess GDF11. Mice were injected with GDF11 secreting cells, an identical model to that used to initially identify the in vivo effects of myostatin. GDF11 exposure in mice induced whole body wasting and profound loss of function in cardiac and skeletal muscle over a 14-day period. Loss of cardiac mass preceded skeletal muscle loss. Cardiac histologic and echocardiographic evaluation demonstrated loss of ventricular muscle wall thickness, decreased cardiomyocyte size, and decreased cardiac function 10 days following initiation of GDF11 exposure. Changes in skeletal muscle after GDF11 exposure were manifest at day 13 and were associated with wasting, decreased fiber size, and reduced strength. Changes in cardiomyocytes and skeletal muscle fibers were associated with activation of SMAD2, the ubiquitin-proteasome pathway and autophagy. Thus, GDF11 over administration in vivo results in cardiac and skeletal muscle loss, dysfunction, and death. Here, serum levels of GDF11 by Western blotting were 1.5-fold increased over controls. Although GDF11 effects in vivo are likely dose, route, and duration dependent, its physiologic changes are similar to myostatin and other Activin receptors ligands. These data support that GDF11, like its other closely related TGF-beta family members, induces loss of cardiac and skeletal muscle mass and function.

Pseudo-spatial coherence resonance in an excitable laser with long delayed feedback.

The effect of noise in an excitable semiconductor laser with feedback is studied in the framework of the spatio-temporal representation of long delayed systems. Propagation, noise-induced creation, and destruction of excitable pulses in the pseudo time are observed. The addition of a variable quantity of noise leads to the occurrence of a phenomenon that we term "pseudo-spatial coherence resonance." A phenomenological model well describes the system and allows for a comparison with the experimental observations. A simple Monte Carlo approach is also introduced and permits to explain the features observed in terms of the key dynamical ingredients of the physical system.

An Intelligent Cooperative Visual Sensor Network for Urban Mobility.

Smart cities are demanding solutions for improved traffic efficiency, in order to guarantee optimal access to mobility resources available in urban areas. Intelligent video analytics deployed directly on board embedded sensors offers great opportunities to gather highly informative data about traffic and transport, allowing reconstruction of a real-time neat picture of urban mobility patterns. In this paper, we present a visual sensor network in which each node embeds computer vision logics for analyzing in real time urban traffic. The nodes in the network share their perceptions and build a global and comprehensive interpretation of the analyzed scenes in a cooperative and adaptive fashion. This is possible thanks to an especially designed Internet of Things (IoT) compliant middleware which encompasses in-network event composition as well as full support of Machine-2-Machine (M2M) communication mechanism. The potential of the proposed cooperative visual sensor network is shown with two sample applications in urban mobility connected to the estimation of vehicular flows and parking management. Besides providing detailed results of each key component of the proposed solution, the validity of the approach is demonstrated by extensive field tests that proved the suitability of the system in providing a scalable, adaptable and extensible data collection layer for managing and understanding mobility in smart cities.

Subclinical pulmonary congestion is prevalent in nephrotic syndrome.

In patients with nephrotic syndrome (NS), the lung is considered an organ protected from the risk of edema. However, data on objectively measured lung water in NS patients is lacking. Here we measured lung water by an ultrasound (US) technique as well as by transthoracic impedance in 42 asymptomatic patients with active NS, in 14 stage G5D CKD patients on chronic hemodialysis, and in 21 healthy individuals. In patients with active NS, the median number of US-B lines (a metric of lung water) after 5 min in a supine position was significantly higher (12; interquartile range: 7-25) compared with that in healthy individuals (4; 2-9) but similar to that in hemodialysis patients (23; 10-39). The difference between NS patients and healthy individuals was significantly amplified (16; 10-35 vs. 4; 2-9) after 60 min of supine resting and significantly attenuated after 5 min of standing (10; 7-25 vs. 3; 1-6). Posture-dependent changes in lung water in patients with active NS were significantly accentuated compared with both hemodialysis patients and healthy individuals. After NS remission, the number of US-B lines was significantly reduced to 5 (4-18) at 5 min and to 6 (5-22) at 60 min approaching the normal range. Lung congestion in patients with active NS was confirmed by transthoracic impedance. Thus, asymptomatic pulmonary congestion is pervasive in patients with NS. A clinical trial is needed to assess the utility of lung US for the management of patients with NS.

Testing Quantum Gravity Induced Nonlocality via Optomechanical Quantum Oscillators.

Several quantum gravity scenarios lead to physics below the Planck scale characterized by nonlocal, Lorentz invariant equations of motion. We show that such nonlocal effective field theories lead to a modified Schrödinger evolution in the nonrelativistic limit. In particular, the nonlocal evolution of optomechanical quantum oscillators is characterized by a spontaneous periodic squeezing that cannot be generated by environmental effects. We discuss constraints on the nonlocality obtained by past experiments, and show how future experiments (already under construction) will either see such effects or otherwise cast severe bounds on the nonlocality scale (well beyond the current limits set by the Large Hadron Collider). This paves the way for table top, high precision experiments on massive quantum objects as a promising new avenue for testing some quantum gravity phenomenology.

Transnasal administration of a combination of dexmedetomidine, midazolam and butorphanol produces deep sedation in New Zealand White rabbits.

OBJECTIVE: To study the sedative and cardiorespiratory effects of transnasal (TN) administration of a combination of dexmedetomidine (DEX), midazolam (MID) and butorphanol (BUT) administered through a nasal catheter to rabbits undergoing diagnostic procedures. STUDY DESIGN: Descriptive cross-sectional experimental study. ANIMALS: Eight healthy New Zealand White rabbit does (12 ± 1 months old, 3.5 ± 0.3 kg). METHODS: DEX (0.1 mg kg(-1)), MID (2 mg kg(-1)) and BUT (0.4 mg kg(-1)) were mixed (DMB) in a syringe and applied to the rabbits' nasopharyngeal mucosa after the accurate catheterization of one nostril. The onset, duration and quality of effects including analgesia were scored using a numeric rating scale of sedation for rabbits. Continuous monitoring of vital parameters was performed via clinical and multiparametric recording. Physiological variables were explored using repeated measures anova for parametric data or Friedman's test for non-parametric data. Tukey's or Dunn's post hoc multiple comparisons test was used depending on normality. The statistical significance was set at p < 0.05. RESULTS: Loss of the righting reflex, deep sedation and profound analgesia ensued simultaneously at 1.4 ± 1.1 minutes after DMB administration. These effects lasted 45 minutes before subsiding into moderate sedation, which lasted for an additional 25 minutes. Residual central nervous system impairment persisted up to 100 minutes. Blood pressure dropped progressively over time by 50%, whereas respiratory frequency decreased by 70%, consistent with moderate hypoxemia and hypercarbia. CONCLUSION AND CLINICAL RELEVANCE: The TN route is a reliable and effective means for administration of DEX, MID and BUT to rabbits. The overall profound sedative effects and analgesic proprieties of the DMB combination can be selectively reversed depending on the needs of the procedure. Oxygen supplementation and careful monitoring are mandatory even in healthy subjects. The DMB protocol should be cautiously used in rabbits with cardiovascular or respiratory deficiencies.

Experimental Robotic Pulmonary Lobectomy With the TELELAP/ALFX System in the Ovine Model.

A new robotic telesurgical device (TELELAP/ALFX) is used for the first time to execute an anatomical pulmonary resection (lobectomy) plus mediastinal lymph node dissection in the ovine model. This integrated operative system has 2 innovative peculiarities: (a) tactile perception (engineered to give the operator a tactile feedback similar to that experienced when handling thoracoscopy instruments) and (b) eye-tracking (immediate synchronization of the surgeon's eyes movements with that of the robotic camera). Herein, we report a lower right pulmonary lobectomy under complete robotic assistance (TELELAP/ALFX). Standard endoscopic staplers were used in all the major maneuvers (bronchial as well as vascular resections and fissural completion) introduced through a utility 4-cm-sized incision. The specimen was placed in an endoscopic retrieval bag and removed through a service minithoracotomy. With the limitations because of interspecies differences in anatomical landmarks, a mediastinal lymph nodal dissection was also completed. The operative time was acceptable (~180 minutes) with blood loss of 100 mL. In conclusion, according to this first all experimental experience we may deem the TELELAP/ALFX system completely apt to perform major anatomic pulmonary resections and the regulatory process to run trials in humans are under way.

The inhibin/activin signalling pathway in human gonadal and adrenal cancers.

The biological function of the inhibin-α subunit (INHA) in gonadal tumorigenesis is different in humans compared with mouse. The INHA subunit is up-regulated in most human ovarian and testicular cancers but knock-out studies in mice showed the INHA subunit is a tumour suppressor with gonadal and adrenal specificity. The INHA subunit is a component of the inhibin/activin signalling pathway, which includes activin receptors ActRIIA/IIB and intracellular Smads-2/3. To resolve the incongruity in function in humans versus mouse, we re-evaluated the inhibin/activin pathway in human gonadal and adrenal cancers using contemporary protein and mRNA expression data for multiple pathway components rather than INHA alone. We used an INHA antibody raised against the N-terminal domain to compare immunoreactivity with the more commonly used antibody raised against the C-terminal domain. This study also described, for the first time, a comprehensive protein expression profile of activin-ßC in reproductive and adrenal cancers, and its effect on a human granulosa cell line, providing evidence for a role in ovarian, testis and adrenal tumour biology. Our data show reduced INHA expression at both protein and mRNA levels, and increased activin signalling in human testicular, ovarian and malignant versus benign forms of adrenal cancer. We also found that activin-C acts as an activin-A antagonist by binding to activin receptor subunits IIA and IIB and modulating the canonical Smad pathway. In conclusion, analysis of the inhibin/activin signalling pathway helps to explain discrepancies arising from studies of only one hormone or subunit and suggests that altered expression of the inhibin and activin subunits is associated with reproductive and adrenal cancer biology.

Front pinning and localized states analogues in long-delayed bistable systems.

Localized structures have been observed in many spatially extended systems of either biological, chemical, or physical nature. Here, we study experimentally front pinning and dissipative localized structures in a delayed optical system based on a bistable semiconductor laser with optoelectronic feedback. We observe that many of the concepts known to apply to spatially localized structures also apply in this context, with specificities related to the lack of reversibility symmetry. Numerical simulations based on purely prototypical modeling reproduce very well the experimental findings, which indicates that the results do not depend on the specific physical system under consideration, but are, on the contrary, very generic features of time delayed systems.

Activin-ß(c) reduces reproductive tumour progression and abolishes cancer-associated cachexia in inhibin-deficient mice.

Activins are involved in the regulation of a diverse range of physiological processes including development, reproduction, and fertility, and have been implicated in the progression of cancers. Bioactivity is regulated by the inhibin α-subunit and by an activin-binding protein, follistatin. The activin-ß(C) subunit was not considered functionally significant in this regard due to an absence of phenotype in knockout mice. However, activin-ß(C) forms heterodimers with activin-ß(A) and activin-C antagonizes activin-A in vitro. Thus, it is proposed that overexpression, rather than loss of activin-ß(C) , regulates activin-A bioactivity. In order to prove biological efficacy, inhibin α-subunit knockout mice (α-KO) were crossed with mice overexpressing activin-ß(C) (ActC++). Deletion of inhibin leads to Sertoli and granulosa cell tumours, increased activin-A, and cancer-associated cachexia. Therefore, cachexia and reproductive tumour development should be modulated in α-KO/ActC++ mice, where excessive activin-A is the underlying cause. Accordingly, a reduction in activin-A, no significant weight loss, and reduced incidence of reproductive tumours were evident in α-KO/ActC++ mice. Overexpression of activin-ß(C) antagonized the activin signalling cascade; thus, the tumourigenic effects of activin-A were abrogated. This study provides proof of the biological relevance of activin-ß(C) . Being a regulator of activin-A, it is able to abolish cachexia and modulate reproductive tumour development in α-KO mice.

Suppurative thyroiditis, a sign of branchiogenic fistula? Lesson based on a case report.

Branchiogenic fistulas are congenital alterations that affect the cervical compartments. Those of the fourth branchial cleft are rarest and can begin late with very serious complications. The suppurative thyroiditis can be a complication of these alterations. We describe a case of 3-year-old girl with high fever, left cervical swelling and increased inflammation indices. The neck ultrasound showed an abscess of the left thyroid lobe and a fluid mass with aerial content in laterocervical region. On MRI, the fluid mass extended from the left piriform sinus to the mediastinum. Fluoroscopy also highlighted a fistolose trait that extended from the left side wall of the esophagus, anteriorly towards the trachea. Treatment of these pathologies must be early and a late diagnosis can put patients' lives at risk.