Amyloidogenicity assessment of transthyretin gene variants.

OBJECTIVE: Hereditary transthyretin-mediated amyloidosis is a treatable condition caused by amyloidogenic variants in the transthyretin-gene resulting in severe peripheral neuropathy or cardiomyopathy. Only about a third of over 130 known variants are clearly pathogenic, most are classified as variants of uncertain significance. A clear delineation of these into pathogenic or non-pathogenic is highly desirable but hampered by low frequency and penetrance. We thus sought to characterize their amylogenic potential by an unbiased in vitro approach. METHODS: Thioflavin T and turbidity assays were used to compare the potential of mammalian cell expressed wt-transthyretin and 12 variant proteins (either variants of uncertain significance, benign, pathogenic) to aggregate and produce amyloid fibrils in vitro. As proof of principle, the assays were applied to transthyretin-Ala65Val, a variant that was newly detected in a family with peripheral neuropathy and amyloid deposits in biopsies. In silico analysis was performed to compare the position of the benign and pathogenic variants. RESULTS: Transthyretin-Ala65Val showed a significantly higher amyloidogenic potential than wt-transthyretin, in both turbidity- and Thioflavin T-assays, comparable to known pathogenic variants. The other eight tested variants did not show an increased amyloidogenic potential. In silico structural analysis further confirmed differences between pathogenic and benign variants in position and interactions. INTERPRETATION: We propose a biochemical approach to assess amyloidogenic potential of transthyretin variants. As exemplified by transthyretin-Ala65Val, data of three assays together with histopathology clearly demonstrates its amyloidogenicity.

Quantum error correction of a qubit encoded in grid states of an oscillator.

The accuracy of logical operations on quantum bits (qubits) must be improved for quantum computers to outperform classical ones in useful tasks. One method to achieve this is quantum error correction (QEC), which prevents noise in the underlying system from causing logical errors. This approach derives from the reasonable assumption that noise is local, that is, it does not act in a coordinated way on different parts of the physical system. Therefore, if a logical qubit is encoded non-locally, we can-for a limited time-detect and correct noise-induced evolution before it corrupts the encoded information1. In 2001, Gottesman, Kitaev and Preskill (GKP) proposed a hardware-efficient instance of such a non-local qubit: a superposition of position eigenstates that forms grid states of a single oscillator2. However, the implementation of measurements that reveal this noise-induced evolution of the oscillator while preserving the encoded information3-7 has proved to be experimentally challenging, and the only realization reported so far relied on post-selection8,9, which is incompatible with QEC. Here we experimentally prepare square and hexagonal GKP code states through a feedback protocol that incorporates non-destructive measurements that are implemented with a superconducting microwave cavity having the role of the oscillator. We demonstrate QEC of an encoded qubit with suppression of all logical errors, in quantitative agreement with a theoretical estimate based on the measured imperfections of the experiment. Our protocol is applicable to other continuous-variable systems and, in contrast to previous implementations of QEC10-14, can mitigate all logical errors generated by a wide variety of noise processes and facilitate fault-tolerant quantum computation.

To catch and reverse a quantum jump mid-flight.

In quantum physics, measurements can fundamentally yield discrete and random results. Emblematic of this feature is Bohr's 1913 proposal of quantum jumps between two discrete energy levels of an atom1. Experimentally, quantum jumps were first observed in an atomic ion driven by a weak deterministic force while under strong continuous energy measurement2-4. The times at which the discontinuous jump transitions occur are reputed to be fundamentally unpredictable. Despite the non-deterministic character of quantum physics, is it possible to know if a quantum jump is about to occur? Here we answer this question affirmatively: we experimentally demonstrate that the jump from the ground state to an excited state of a superconducting artificial three-level atom can be tracked as it follows a predictable 'flight', by monitoring the population of an auxiliary energy level coupled to the ground state. The experimental results demonstrate that the evolution of each completed jump is continuous, coherent and deterministic. We exploit these features, using real-time monitoring and feedback, to catch and reverse quantum jumps mid-flight-thus deterministically preventing their completion. Our findings, which agree with theoretical predictions essentially without adjustable parameters, support the modern quantum trajectory theory5-9 and should provide new ground for the exploration of real-time intervention techniques in the control of quantum systems, such as the early detection of error syndromes in quantum error correction.

Fault-tolerant detection of a quantum error.

A critical component of any quantum error-correcting scheme is detection of errors by using an ancilla system. However, errors occurring in the ancilla can propagate onto the logical qubit, irreversibly corrupting the encoded information. We demonstrate a fault-tolerant error-detection scheme that suppresses spreading of ancilla errors by a factor of 5, while maintaining the assignment fidelity. The same method is used to prevent propagation of ancilla excitations, increasing the logical qubit dephasing time by an order of magnitude. Our approach is hardware-efficient, as it uses a single multilevel transmon ancilla and a cavity-encoded logical qubit, whose interaction is engineered in situ by using an off-resonant sideband drive. The results demonstrate that hardware-efficient approaches that exploit system-specific error models can yield advances toward fault-tolerant quantum computation.

Deterministic Remote Entanglement of Superconducting Circuits through Microwave Two-Photon Transitions.

Large-scale quantum information processing networks will most probably require the entanglement of distant systems that do not interact directly. This can be done by performing entangling gates between standing information carriers, used as memories or local computational resources, and flying ones, acting as quantum buses. We report the deterministic entanglement of two remote transmon qubits by Raman stimulated emission and absorption of a traveling photon wave packet. We achieve a Bell state fidelity of 73%, well explained by losses in the transmission line and decoherence of each qubit.

A CNOT gate between multiphoton qubits encoded in two cavities.

Entangling gates between qubits are a crucial component for performing algorithms in quantum computers. However, any quantum algorithm must ultimately operate on error-protected logical qubits encoded in high-dimensional systems. Typically, logical qubits are encoded in multiple two-level systems, but entangling gates operating on such qubits are highly complex and have not yet been demonstrated. Here we realize a controlled NOT (CNOT) gate between two multiphoton qubits in two microwave cavities. In this approach, we encode a qubit in the high-dimensional space of a single cavity mode, rather than in multiple two-level systems. We couple two such encoded qubits together through a transmon, which is driven by an RF pump to apply the gate within 190 ns. This is two orders of magnitude shorter than the decoherence time of the transmon, enabling a high-fidelity gate operation. These results are an important step towards universal algorithms on error-corrected logical qubits.

Intramyocardial inflammation predicts adverse outcome in patients with cardiac AL amyloidosis.

AIMS: To evaluate the influence of endomyocardial biopsy (EMB)-proven intramyocardial inflammation on mortality in patients with cardiac transthyretin amyloid (ATTR) or amyloid light-chain (AL) amyloidosis. METHODS AND RESULTS: We included 54 consecutive patients (mean age 68.83 ± 9.59 years; 45 men) with EMB-proven cardiac amyloidosis. We followed up patients from first diagnostic biopsy to as long as 36 months (mean 11.5 ± 12 months) and compared their outcome with information on all-cause mortality with or without proof of inflammation on EMB. Intramyocardial inflammation was assessed by quantitative immunohistology. Patients suffering from amyloidosis revealed a significant poor prognosis with proof of intramyocardial inflammation in contrast to those without inflammation (log-rank P = 0.019). Re-grouping of patients indicated AL amyloidosis to have a significant impact on all-cause mortality (log-rank P = 0.012). The detailed subgroup analysis showed that patients suffering from AL amyloidosis with intramyocardial inflammation have a significantly worse prognosis compared with AL amyloidosis without inflammation and ATTR with or without inflammation, respectively (log-rank P = 0.014, contingency Fisher's exact test, P = 0.008). CONCLUSION: Our study reports for the first time a high incidence (48.1%) of intramyocardial inflammation in a series of patients with EMB-proven cardiac amyloidosis and could show that in patients with AL amyloidosis, intramyocardial inflammation correlated significantly with increased mortality. Our data have a direct clinical impact because one can hypothesize that additional immunomodulating/anti-inflammatory treatment regimens in patients with biopsy-proven inflammation of heart muscle tissue could be beneficial for patients suffering from cardiac AL amyloidosis.

Age-dependent physiological dynamics in acid-base balance, electrolytes, and blood metabolites in growing goats.

There is a paucity of published data reporting acid-base equilibrium in goats, and no information is available on how the acid-base complexity changes when suckling goat kids become ruminants. The aims of this study were to evaluate young healthy goats for age-related changes in serum proteins, metabolites, and electrolytes; differences in results when the Henderson-Hasselbalch equation or strong ion approaches were used were also assessed. To assess biological variability and reproducibility, two consecutive long-term studies, each lasting from the 6th to 56th week of life (wl), were performed in 15 (Study 1) and 10 (Study 2) animals. Blood gas analysis, serum biochemical analysis, and electrophoresis were performed on venous blood, and acid-base information was obtained using the traditional Henderson-Hasselbalch approach, Stewart's strong ion model, and Constable's simplified strong ion model. In all goats within the first 4-5 months, serum concentrations of glucose, l-lactate, and inorganic phosphate decreased significantly, while serum concentrations of total protein, albumin, and gamma globulin increased. Consequently, nonvolatile weak acids (Atot Alb and Atot TP) increased. At the end of this 'adaptation period', i.e. when milk was replaced by purely plant-based food, significantly lower bicarbonate and base excess values were accompanied by blood pH that shifted towards acidosis. Electrolytes (Na+, K+, Ca2+, and Cl-), anion gap, strong ion difference, and strong ion gap did not show age-dependent trends. In conclusion, somatic growth and development of gastro-intestinal fermentation in growing goats act as complex sources of physiological variability on acid-base equilibrium that was not reflected by the Henderson-Hasselbalch equation only.

Effect of Parachlamydia acanthamoebae on pulmonary function parameters in a bovine respiratory model.

The aim of this study was to evaluate pulmonary dysfunction induced by experimental infection with Parachlamydia acanthamoebae in calves. Intrabronchial inoculation with P. acanthamoebae was performed in 31 calves aged 2-3 months old at two different challenge doses of 10(8) and 10(10) inclusion-forming units (IFU) per animal. Control animals received heat inactivated bacteria. The effects on pulmonary gas exchange were determined by arterial blood gas analysis and haemoximetry during the 7 days post inoculation (DPI). For pulmonary function testing (PFT), impulse oscillometry, capnography, and measurement of O2 uptake were undertaken in spontaneously breathing animals 7 and 3 days before inoculation and were repeated until 10 DPI. In the early phase after challenge (1-3 DPI), mild hypoxaemia occurred, which was accompanied by a significant reduction in both tidal and alveolar volumes (each related to bodyweight, BW). In parallel, expiratory flow rate and specific ventilation (i.e. minute ventilation related to O2 uptake) were significantly increased. Minute and alveolar ventilations (each related to metabolic BW) increased significantly due to higher respiratory rates, lasting until 4 and 5 DPI, respectively. Oxygen uptake was slightly reduced during the first 2 days after challenge, but increased significantly during the recovery phase, from 4 to 8 DPI. No deterioration in respiratory mechanics or acid-base balance was observed. Respiratory infection with 10(10) IFU P. acanthamoebae per calf induced mild respiratory dysfunction, mainly characterised by hypoxaemia. The study's findings do not indicate severe pathophysiological consequences of P. acanthamoebae infection on pulmonary function in the bovine host.

Low levels of C-peptide have clinical significance for established Type 1 diabetes.

AIM: To determine whether the low C-peptide levels (< 50 pmol/l) produced by the pancreas for decades after onset of Type 1 diabetes have clinical significance. METHODS: We evaluated fasting C-peptide levels, duration of disease and age of onset in a large cross-sectional series (n = 1272) of people with Type 1 diabetes. We then expanded the scope of the study to include the relationship between C-peptide and HbA1c control (n = 1273), as well as diabetic complications (n = 324) and presence of hypoglycaemia (n = 323). The full range of C-peptide levels was also compared with 1,5-Anhydroglucitol, a glucose responsive marker. RESULTS: C-peptide levels declined for decades after diagnosis, and the rate of decline was significantly related to age of onset (P < 0.0001), after adjusting for disease duration. C-peptide levels > 10 pmol/l were associated with protection from complications (e.g. nephropathy, neuropathy, foot ulcers and retinopathy; P = 0.03). Low C-peptide levels were associated with poor metabolic control measured by HbA1c (P < 0.0001). Severe hypoglycaemia was associated with the lowest C-peptide levels compared with mild (P = 0.049) or moderate (P = 0.04) hypoglycaemia. All levels of measurable C-peptide were responsive to acute fluctuations in blood glucose levels as assessed by 1,5-Anhydroglucitol (P < 0.0001). CONCLUSIONS: Low C-peptide levels have clinical significance and appear helpful in characterizing groups at-risk for faster C-peptide decline, complications, poorer metabolic control and severe hypoglycaemia. Low C-peptide levels may be a biomarker for characterizing at-risk patients with Type 1 diabetes.

Relationship between clinical signs and results of impulse oscillometry in pigs originating from the field.

The aim of the present study was to assess the contribution of clinical data to the variability of impulse oscillometric test results observed previously by Püllen et al. (2014). Fifty-eight German hybrid pigs from 29 different herds with unknown respiratory status were examined in the context of routine diagnostics as part of herd health service. Routine clinical examination was extended to a total set of 29 parameters, representing detailed clinical signs of the respiratory system, and to lung function testing applying the impulse oscillometry system (IOS). The resulting linear relationship between clinical data and variables of pulmonary mechanics had a mean r(2) of 0.52. Clinical parameters predominantly representing the lower respiratory tract closely correlated with established impulse oscillometric indices reflecting peripheral airways. Because of a restricted relationship between pulmonary functional disorders and clinical data, additional diagnostic methods are required to reveal the proportion of variance undefined by clinical examination.

Gastrointestinal ALκ amyloidosis presenting as protein-losing enteropathy correctly diagnosed by immunohistochemistry using amyloid-type-specific antibodies.

A 54-year-old man presented with protein-losing enteropathy. Biopsies from the stomach, duodenum, ileum and colon showed deposits of amyloid. The bone marrow showed plasmacytosis. After an initial misdiagnosis of AA amyloid, a revised diagnosis of ALκ amyloidosis was made at an expert referral laboratory. Care must be taken in the use of antibodies and proper controls in the performance and interpretation of immunohistochemistry for amyloidosis. A wide panel of amyloid-type-specific antibodies must be used and interpreted in comparative mode to avoid misdiagnosis.

Laparoscopic placement of a tined lead electrode on the pudendal nerve with urodynamic monitoring of bladder function during electrical stimulation: an acute experimental study in healthy female pigs.

PURPOSE: The aim of this study was to develop a method for standard laparoscopic access to the pudendal nerve in pigs to implant an electrode for chronic neuromodulation studies. METHODS: Using routine laparoscopic surgical techniques, the pudendal nerve was located in 10 female pigs using standardized anatomical landmarks. A tined lead electrode was placed in parallel to the exposed pudendal nerve, and acute unilateral electrical stimulation was performed consecutively on both pudendal nerves. Bladder pressure and perineal skeletal muscle response was monitored during stimulation. RESULTS: Standard access to the pudendal nerve was successfully established in the pig model with surgical times of approximately 45 minutes for bilateral electrode placement. Acute unilateral stimulation did not evoke bladder responses but resulted in reliable stimulation-dependent activity of the perineal skeletal muscles. The structural integrity of the pudendal nerves was confirmed in all cases. CONCLUSIONS: These results illustrate the effectiveness of laparoscopy for standardised, safe nerve localisation and electrode implantation at the pudendal nerve in pigs. Laparoscopic implantation represents an alternative approach for performing electrode implantation under optical guidance versus the standard approach of percutaneous, neuro-physiological monitored implantation. In the future, pudendal neuromodulation may be used as a supplement to sacral neuromodulation or as a standalone therapeutic approach, depending on the underlying bladder dysfunction.

Evaluation of impulse oscillometry in pigs of unknown disease status originating from the field.

The aim of this study was to assess impulse oscillometry as a method to characterise lung function in 58 German hybrid pigs from 29 different herds of unknown respiratory status. The variability of repeated lung function measurements increased significantly after the sixth run and therefore the average of the first six runs was used for analysis. The presence of peripheral respiratory alterations in some pigs was indicated by the negative frequency dependence of the 95th percentile of respiratory resistance (Rrs), with highest values at 3 Hz and the sharp drop of respiratory reactance (Xrs) across the whole frequency range (3-15 Hz). Respiratory resistance and reactance were negatively correlated. Reactance area was correlated with (1) Rrs at 3, 5 and 10 Hz; (2) Xrs at 3, 5, 10 and 15 Hz; (3) the frequency dependence of resistance compared between 3 and 5 Hz (R3-R5), 5 and 10 Hz (R5-R10), and 5 and 15 Hz (R5-R15); and (4) tidal volume. High repeatability and low intra-individual variability of impulse oscillometry indicate that this method is a promising tool for advanced characterisation of the pulmonary system of pigs and has potential for use for herd health monitoring.

Serum amyloid A: expression throughout human ovarian folliculogenesis and levels in follicular fluid of women undergoing controlled ovarian stimulation.

BACKGROUND: Serum amyloid A (SAA) is an acute phase protein expressed primarily in the liver in response to various injuries and inflammatory stimuli and is recognized as a modulator of inflammation. Ovarian reproductive functions including folliculogenesis and ovulation use inflammatory processes; thus, studying SAA in this context is of interest. OBJECTIVES: We investigated the expression and localization of SAA in ovarian developing follicles and its levels in follicular fluids. METHODS AND PARTICIPANTS: Nonradioactive in situ hybridization and immunohistochemical staining were applied on ovarian paraffin tissue sections. ELISA and RT-PCR were applied on follicular aspirates and blood samples from women undergoing controlled ovarian stimulation for in vitro fertilization. RESULTS: Expression of SAA mRNA and protein was found in follicular cells at all stages of follicular development, from primordial and primary follicles through antral follicles and corpora lutea. Expression was observed in granulosa, theca and luteal cells, and oocytes. Expression of SAA was also found in granulosa cells recovered from follicular aspirates. The SAA protein was detected in follicular fluids. Its levels were somewhat lower than in peripheral blood with strong correlation between the two compartments and with significant correlation with patient's body mass index. High follicular fluid SAA levels were associated with reduced pregnancy rate. CONCLUSIONS: SAA is locally produced in ovarian developing follicles and is a constituent of follicular fluids, suggesting its role within the follicular environment. Elevated follicular SAA levels are associated with decreased pregnancy rate and may signify lower reproductive performance.