Metabolic shift underlies recovery in reversible infantile respiratory chain deficiency.

Reversible infantile respiratory chain deficiency (RIRCD) is a rare mitochondrial myopathy leading to severe metabolic disturbances in infants, which recover spontaneously after 6-months of age. RIRCD is associated with the homoplasmic m.14674T>C mitochondrial DNA mutation; however, only ~ 1/100 carriers develop the disease. We studied 27 affected and 15 unaffected individuals from 19 families and found additional heterozygous mutations in nuclear genes interacting with mt-tRNAGlu including EARS2 and TRMU in the majority of affected individuals, but not in healthy carriers of m.14674T>C, supporting a digenic inheritance. Our transcriptomic and proteomic analysis of patient muscle suggests a stepwise mechanism where first, the integrated stress response associated with increased FGF21 and GDF15 expression enhances the metabolism modulated by serine biosynthesis, one carbon metabolism, TCA lipid oxidation and amino acid availability, while in the second step mTOR activation leads to increased mitochondrial biogenesis. Our data suggest that the spontaneous recovery in infants with digenic mutations may be modulated by the above described changes. Similar mechanisms may explain the variable penetrance and tissue specificity of other mtDNA mutations and highlight the potential role of amino acids in improving mitochondrial disease.

Design and Preclinical Development of TransCon PTH, an Investigational Sustained-Release PTH Replacement Therapy for Hypoparathyroidism.

Hypoparathyroidism (HP) is a condition of parathyroid hormone (PTH) deficiency leading to abnormal calcium and phosphate metabolism. The mainstay of therapy consists of vitamin D and calcium supplements, as well as adjunct Natpara (PTH(1-84)). However, neither therapy optimally controls urinary calcium (uCa) or significantly reduces the incidence of hypercalcemia and hypocalcemia. TransCon PTH, a sustained-release prodrug of PTH(1-34) in development for the treatment of HP, was designed to overcome these limitations. To determine the pharmacokinetics and pharmacodynamics of TransCon PTH, single and repeat s.c. dose studies were performed in rats and monkeys. TransCon PTH demonstrated a half-life of 28 and 34 hours in rats and monkeys, respectively. After repeated dosing, an infusion-like profile of the released PTH, characterized by low peak-to-trough levels, was obtained in both species. In intact rats and monkeys, daily subcutaneous administration of TransCon PTH was associated with increases in serum calcium (sCa) levels and decreases in serum phosphate levels (sP). In monkeys, at a single dose of TransCon PTH that increased sCa levels within the normal range, a concurrent decrease in uCa excretion was observed. In 4-week repeat-dose studies in intact rats and monkeys, uCa excretion was comparable to controls across all dose levels despite increases in sCa levels. Further, in a rat model of HP, TransCon PTH normalized sCa and sP levels 24 hours per day. This was in contrast to only transient trends toward normalization of sCa and sP levels with an up to 6-fold higher molar dose of PTH(1-84). After repeated dosing to HP rats, uCa excretion transiently increased, corresponding to increases in sCa above normal range, but at the end of the treatment period, uCa excretion was generally comparable to sham controls. TransCon PTH was well tolerated and the observed pharmacokinetics and pharmacodynamics were in line with the expected action of physiological replacement of PTH. © 2019 American Society for Bone and Mineral Research.

A quantum-dot heat engine operating close to the thermodynamic efficiency limits.

Cyclical heat engines are a paradigm of classical thermodynamics, but are impractical for miniaturization because they rely on moving parts. A more recent concept is particle-exchange (PE) heat engines, which uses energy filtering to control a thermally driven particle flow between two heat reservoirs1,2. As they do not require moving parts and can be realized in solid-state materials, they are suitable for low-power applications and miniaturization. It was predicted that PE engines could reach the same thermodynamically ideal efficiency limits as those accessible to cyclical engines3-6, but this prediction has not been verified experimentally. Here, we demonstrate a PE heat engine based on a quantum dot (QD) embedded into a semiconductor nanowire. We directly measure the engine's steady-state electric power output and combine it with the calculated electronic heat flow to determine the electronic efficiency η. We find that at the maximum power conditions, η is in agreement with the Curzon-Ahlborn efficiency6-9 and that the overall maximum η is in excess of 70% of the Carnot efficiency while maintaining a finite power output. Our results demonstrate that thermoelectric power conversion can, in principle, be achieved close to the thermodynamic limits, with direct relevance for future hot-carrier photovoltaics10, on-chip coolers or energy harvesters for quantum technologies.

Large nuclear spin polarization in gate-defined quantum dots using a single-domain nanomagnet.

The electron-nuclei (hyperfine) interaction is central to spin qubits in solid state systems. It can be a severe decoherence source but also allows dynamic access to the nuclear spin states. We study a double quantum dot exposed to an on-chip single-domain nanomagnet and show that its inhomogeneous magnetic field crucially modifies the complex nuclear spin dynamics such that the Overhauser field tends to compensate external magnetic fields. This turns out to be beneficial for polarizing the nuclear spin ensemble. We reach a nuclear spin polarization of ≃50%, unrivaled in lateral dots, and explain our manipulation technique using a comprehensive rate equation model.

Identification of the western tarnished plant bug (Lygus hesperus) olfactory co-receptor Orco: expression profile and confirmation of atypical membrane topology.

Lygus hesperus (western tarnished plant bug) is an agronomically important pest species of numerous cropping systems. Similar to other insects, a critical component underlying behaviors is the perception and discrimination of olfactory cues. Consequently, the molecular basis of olfaction in this species is of interest. To begin to address this issue, we utilized homology-based PCR as a commonly accepted abbreviation but if necessary it is polymerase chain reaction methods to identify the L. hesperus olfactory receptor co-receptor (Orco) ortholog, a receptor that has been shown to be essential for olfaction. The L. hesperus Orco (LhOrco) shares significant sequence homology with known Orco proteins in other insects. Parallel experiments using the sympatric sister species, Lygus lineolaris (tarnished plant bug), revealed that the Lygus Orco gene was completely conserved. Surprisingly, a majority of the membrane topology prediction algorithms used in the study predicted LhOrco to have both the N and C terminus intracellular. In vitro immunofluorescent microscopy experiments designed to probe the membrane topology of transiently expressed LhOrco, however, refuted those predictions and confirmed that the protein adopts the inverted topology (intracellular N terminus and an extracellular C terminus) characteristic of Orco proteins. RT-PCR analyses indicated that LhOrco transcripts are predominantly expressed in adult antennae and to a lesser degree in traditionally nonolfactory chemosensory tissues of the proboscis and legs. Expression is not developmentally regulated because transcripts were detected in all nymphal stages as well as eggs. Taken together, the results suggest that LhOrco likely plays a critical role in mediating L. hesperus odorant perception and discrimination.

Penetrative and dislodgeable residue characteristics of 14C-insecticides in apple fruit.

Infinite- and finite-dose laboratory experiments were used to study the penetrative and dislodgeable residue characteristics of (14)C-insecticides in apple fruit. The differences in dislodgeable and penetrated residues of three radiolabeled insecticides ((14)C-thiamethoxam, (14)C-thiacloprid, and (14)C-indoxacarb), applied in aqueous solution with commercial formulations, were determined after water and methanol wash extractions. The rate of sorption and extent of penetration into the fruit cuticles and hypanthium of two apple cultivars were measured after 1, 6, and 24 h of treatment exposure, using radioactivity quantification methods. For all three compounds, 97% or more of the treatment solutions were found on the fruit surface as some form of non-sorbed residues. For indoxacarb, sorption into the epicuticle was rapid but desorption into the fruit hypanthium was delayed, indicative of a lipophilic penetration pathway. For the neonicotinoids, initial cuticular penetration was slower but with no such delay in desorption into the hypanthium.

Imidacloprid in melon guttation fluid: a potential mode of exposure for pest and beneficial organisms.

ELISA techniques were used to detect imidacloprid in guttation fluid of young cantaloupe plants in Arizona. Imidacloprid was detected at up to 4.1 microg/ml (ppm) in a coincidental guttation collection 3 d after a top label rate soil application and at 37 microg/ml one d after a separate top label rate soil application study. These imidacloprid titers exceed reported median oral toxicities for several insect species by factors of 10 or more. Pesticides in guttation fluid are a relatively unexplored route of exposure for both pest and beneficial insects, and could represent an important risk for both of these groups in guttation-prone environments.

A nanoscale standard for the Seebeck coefficient.

The Seebeck coefficient, a key parameter describing a material's thermoelectric performance, is generally difficult to measure, and no intrinsic calibration standard exists. Quantum dots and single electron tunneling devices with sharp transmission resonances spaced by many kT have a material-independent Seebeck coefficient that depends only on the electronic charge and the average device temperature T. Here we propose the use of a quantum dot to create an intrinsic, nanoscale standard for the Seebeck coefficient and discuss its implementation.

Plum curculio (Coleoptera: Curculionidae) adult mortality and associated fruit injury after exposure to field-aged insecticides on tart cherry branches.

Plum curculio, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), adults were exposed to field-aged residues of thiamethoxam, acetamiprid, thiacloprid, indoxacarb, or azinphos-methyl on tart cherry, Prunus cerasus L. variety Montmorency. At 1, 3, 7, and 14 d postapplication, fruit were sampled for chemical residues, and bioassays were used to assess beetle mortality and plant tissue injury. Azinphos-methyl had lethal activity within 1 d of exposure at all postapplication intervals and significant fruit protection extended to 14 d postapplication. All of the neonicotinoids had lethal activity at 3 d posttreatment, with acetamiprid activity extending to 7 d. Antifeedant and oviposition deterrent effects were seen with thiamethoxam and thiacloprid; damage incidence was significantly reduced in the absence of significant beetle mortality or intoxication. Thiamethoxam and acetamiprid penetrated into leaf and fruit tissue and were detected in the interior tissues at 14 d postapplication, but interior thiacloprid residues were not detected after day 1. Indoxacarb provided some fruit protection out to 7 d postapplication, and 14-d-old residues intoxicated beetles, but the slow action of this compound allowed significant damage to occur before beetles were incapacitated. Indoxacarb was only detected as a surface residue after the first day postapplication. These data on the plant-insect-chemistry interactions will support use and management decisions as compounds with acute contact activity are phased out.

Tumor-reactive CD8+ T-cell responses after vaccination with NY-ESO-1 peptide, CpG 7909 and Montanide ISA-51: association with survival.

Peptide-based vaccines have led to the induction of antigen-specific CD8(+) T-cell responses in patients with NY-ESO-1 positive cancers. However, vaccine-induced T-cell responses did not generally correlate with improved survival. Therefore, we tested whether a synthetic CpG 7909 ODN (deoxycytidyl-deoxyguanosin oligodeoxy-nucleotides) mixed with NY-ESO-1 peptide p157-165 and incomplete Freund's adjuvants (Montanide(R) ISA-51) led to enhanced NY-ESO-1 antigen-specific CD8(+) immune responses in patients with NY-ESO-1 or LAGE-1 expressing tumors. Of 14 HLA-A2+ patients enrolled in the study, 5 patients withdrew prematurely because of progressive disease and 9 patients completed 1 cycle of immunization. Nine of 14 patients developed measurable and sustained antigen-specific CD8(+) T-cell responses: Four had detectable CD8+ T-cells against NY-ESO-1 after only 2 vaccinations, whereas 5 patients showed a late-onset but durable induction of NY-ESO-1 p157-165 specific T-cell response during continued vaccination after 4 months. In 6 patients, vaccine-induced antigen-specific T-cells became detectable ex vivo and reached frequencies of up to 0.16 % of all circulating CD8(+) T-cells. Postvaccine T-cell clones were shown to recognize and lyse NY-ESO-1 expressing tumor cell lines in vitro. In 6 of 9 patients developing NY-ESO-1-specific immune responses, a favorable clinical outcome with overall survival times of 43+, 42+, 42+, 39+, 36+ and 27+ months, respectively, was observed.

Curative activity of insecticides against plum curculio (Coleoptera: Curculionidae) in tart cherries.

Tart cherry, Prunus cerasus L. variety Montmorency, fruit were infested with plum curculio, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), and treated with insecticides to target late instars, neonates, and eggs. The organophosphates azinphos-methyl and phosmet and the neonicotinoid thiamethoxam reduced larval emergence rates by >90% for all life stage targets; after >30 d, few surviving larvae were found inside fruit. Acetamiprid and thiacloprid also had curative activity and yielded >75% reductions in emergence and few surviving larvae in the fruit after >30 d. The juvenile hormone analog pyriproxyfen reduced larval emergence, but 66% of fruit that was treated to target late-instars still had live larvae inside of them after >30 d. Novaluron, chlorantraniliprole, and esfenvalerate had no curative activity. Indoxacarb had limited curative activity, and all targeted life stages had larval emergence. Internal and external residues were analyzed and are discussed in relation to their penetration and curative potential. The curative activity of azinphos-methyl has played an important role in meeting federal standards for infestation-free tart cherries at processing. Regulatory changes are eliminating the use of this compound, and new integrated pest management programs for plum curculio will need to address the loss of azinphos-methyl's curative activity.

Measuring temperature gradients over nanometer length scales.

When a quantum dot is subjected to a thermal gradient, the temperature of electrons entering the dot can be determined from the dot's thermocurrent if the conductance spectrum and background temperature are known. We demonstrate this technique by measuring the temperature difference across a 15 nm quantum dot embedded in a nanowire. This technique can be used when the dot's energy states are separated by many kT and will enable future quantitative investigations of electron-phonon interaction, nonlinear thermoelectric effects, and the efficiency of thermoelectric energy conversion in quantum dots.

Novel life stage targets against plum curculio, Conotrachelus nenuphar (Herbst), in apple integrated pest management.

In this study the authors employed the plant-insect-chemistry (PIC) triad to investigate two novel life stage targets against the plum curculio (PC), Conotrachelus nenuphar (Herbst), in apple integrated pest management (IPM). Laboratory treated apple bioassays were used to determine if the insect growth regulator (IGR) insecticides novaluron and tebufenozide have physiological effects on PC larvae following adult exposure. Curative activity bioassays were conducted for IGR, neonicotinoid, oxidiazine and organophosphate insecticides on PC larvae post-infestation, and fruit penetration profiles of insecticides were developed. The results revealed that novaluron exhibits activity on PC larvae via vertical transmission following exposure of mated females to treated substrate. Surface treatments of azinphos-methyl, thiacloprid and thiamethoxam to preinfested fruit resulted in significant reductions in larval emergence. For all compounds, 50% or more of the total recovered active ingredient was from apple skin, and for azinphos-methyl, indoxacarb and thiamethoxam it was greater than 80%. For azinphos-methyl, novaluron, methoxyfenozide and thiacloprid, however, active ingredient was recovered from every section of the apple, from skin to core. Implications for twenty-first century IPM are discussed.

Pyriproxyfen activates reproduction in prediapause northern strain plum curculio (Conotrachelus nenuphar Herbst).

Field-collected, prediapause northern strain plum curculio, Conotrachelus nenuphar (Herbst), adults were treated with the juvenile hormone analogue pyriproxyfen to assess effects on reproductive development. Adults of this pest have an obligate winter reproductive diapause and do not reach reproductive maturity until after spring emergence. Topical (1.0 microL) doses of 10, 1.0 and 0.1 microg microL(-1) pyriproxyfen induced oocyte development and reproductive maturation in all treated females as assessed by dissection. There was no increased mortality in treated beetles, and control insects showed no reproductive maturation. Treatment of prediapause males and females with 1.0 microg microL(-1) of topical pyriproxyfen or exposure to residues on fruit induced successful egg laying and F1 emergence; F1 pupation success in topical treatments and residue exposure was 47 and 59% respectively. Filial adults require re-exposure to initiate reproductive development. Treatment protocols with pyriproxyfen will allow researchers to culture the northern strain, instead of relying exclusively on the non-diapausing southern strain.

Vaccination with NY-ESO-1 protein and CpG in Montanide induces integrated antibody/Th1 responses and CD8 T cells through cross-priming.

The use of recombinant tumor antigen proteins is a realistic approach for the development of generic cancer vaccines, but the potential of this type of vaccines to induce specific CD8(+) T cell responses, through in vivo cross-priming, has remained unclear. In this article, we report that repeated vaccination of cancer patients with recombinant NY-ESO-1 protein, Montanide ISA-51, and CpG ODN 7909, a potent stimulator of B cells and T helper type 1 (Th1)-type immunity, resulted in the early induction of specific integrated CD4(+) Th cells and antibody responses in most vaccinated patients, followed by the development of later CD8(+) T cell responses in a fraction of them. The correlation between antibody and T cell responses, together with the ability of vaccine-induced antibodies to promote in vitro cross-presentation of NY-ESO-1 by dendritic cells to vaccine-induced CD8(+) T cells, indicated that elicitation of NY-ESO-1-specific CD8(+) T cell responses by cross-priming in vivo was associated with the induction of adequate levels of specific antibodies. Together, our data provide clear evidence of in vivo cross-priming of specific cytotoxic T lymphocytes by a recombinant tumor antigen vaccine, underline the importance of specific antibody induction for the cross-priming to occur, and support the use of this type of formulation for the further development of efficient cancer vaccines.

Use of residue profile analysis to identify modes of insecticide activity contributing to control of plum curculio in apples.

Residue profile analysis techniques were developed, along with laboratory and field-based bioassays to describe the modes of insecticidal activity responsible for the control of the plum curculio, Conotrachelus nenuphar (Herbst), in apples (Malus spp.). Adult plum curculios were treated in laboratory topical bioassays to determine acute contact activity and lethal time for five insecticides. Azinphosmethyl had the highest levels of toxicity and shortest lethal time values, followed by the neonicotinoids thiacloprid, thiamethoxam, and imidacloprid, whereas indoxacarb had the highest LD50 and LT50 values for topical exposure. Field-based residual activity bioassays assessed adult mortality, and fruit and leaf injury from plum curculio exposed to 4 h, 7 d, and 14 d field-aged residues. All compounds caused significant levels of mortality to plum curculio when adults were exposed to fruit clusters 4 h post-application. Thiacloprid, thiamethoxam, and imidacloprid showed oviposition deterrence, antifeedant, and repellency effects in the 7- and/or 14 d residual bioassays and protected fruit in the absence of significant lethal activity. Indoxacarb maintained lethal activity throughout the study intervals, with the incidence of plum curculio feeding, suggesting that ingestion is an important mode of entry. For the neonicotinoids thiacloprid, thiamethoxam and imidacloprid plum curculio mortality was highly correlated with fruit and leaf surface residues. As surface residues declined, sublethal effects such as oviposition deterrence and antifeedant effect remained. The value of the plant-insect-chemistry triad model for describing the temporal dimensions of insecticidal modes of activity and understanding a compound's critical performance characteristics is discussed.

Prednisolone-induced changes in dystrophic skeletal muscle.

Although glucocorticoids delay the progression of Duchenne muscular dystrophy (DMD) their mechanism of action is unknown. Skeletal muscle gene expression profiles of mdx mice, an animal model of DMD, treated with prednisolone were compared with control mice at 1 and 6 wk. Of the 89 early differentially regulated genes and ESTs, delta-sarcoglycan, myosin Va, FK506-binding protein 51 (FKBP51), the potassium channel regulator potassium inwardly-rectifying channel Isk-like (IRK2) and ADAM 10 were overexpressed, whereas growth hormone-releasing hormone receptor (GHRHR) and Homer-2 were underexpressed. The 58 late differentially overexpressed genes included kallikreins (13, 16, and 26), FKBP51, PI3K alpha regulatory subunit, and IGFBP6, while underexpressed genes included NeuroD and nicotinic cholinergic receptor gamma. At both time points, overexpression of a cohort of genes relating to metabolism and proteolysis was apparent, alongside the differential expression of genes relating to calcium metabolism. Treatment did not increase muscle regeneration, reduce the number of infiltrating macrophages, or alter utrophin expression or localization. However, in the treated mdx soleus muscle, the percentage of slow fibers was significantly lower compared with untreated controls after 6 wk of treatment. These results show that glucocorticoids confer their benefit to dystrophic muscle in a complex fashion, culminating in a switch to a more normal muscle fiber type.

Reproductive development of northern and southern strains of plum curculio (Coleoptera: Curculionidae).

Laboratory-reared southern and field-collected northern strains of plum curculio, Conotrachelles nenuphar (Herbst), were sampled to examine the relationship between degree-day (DD) accumulation and female reproductive development, as measured by mating status, oocyte size, and number of oocytes. The overall goal was to generate an objective degree-day model for predicting damage potential that could be applied to various host commodities rather than relying on separate biofix models for each crop. Adult beetles were dissected to measure mating status, maximum oocyte size, and number of oocytes. Southern strain beetles reared at 25 degrees C initiated mating 9 d after eclosion and did not require mating to induce oocyte development. By 20 d posteclosion, unmated females had significantly higher egg loads compared with mated females of the same age. Logistic regression analysis suggests that southern and northern strain beetles had a stable maximum oocyte length of 62 and 72 microm, respectively. Northern strain females mated after overwintering; with approximately 95% of the female population mated after 134 DD (base 10 degrees C), which is before fruit set in many host crops. Oocyte size was the only measured parameter of field reproductive progress that could be linked with confidence to degree-day accumulation. The other two parameters do not share an exclusive relationship with degree-days. Rapid assessment of field-caught female reproductive status could assist in determining the potential for plum curculio damage in high-value commodities and allow for more informed control decisions.

Reassessment of the role and utility of wind in suppression of mosquito (Diptera: Culicidae) host finding: stimulus dilution supported over flight limitation.

In a Central Michigan wetland setting, with abundant Aedes vexans (Meigen), Anopheles walkeri (Theobald), and Coquillettidea perturbans (Walker), electric fan-generated wind strongly reduced mosquito catches in Centers for Disease Control and Prevention light traps releasing carbon dioxide at 650 or 1,950 ml/min. This relationship was negatively logarithmic over the range of velocities tested (0 m/s to 3.7 m/s; 0-8.3 mph) and closely matched the theoretical dilution function predicted to occur with increasing wind flows over a constantly releasing point source of volatile chemical. Tripling the carbon dioxide release rate consistently doubled the number of mosquitoes caught across the range of wind velocities tested. Neither applied wind velocity nor carbon dioxide release rate was correlated with mosquito body mass across the range of wind velocities tested. Collectively, these data support the conceptual model that applied wind diminishes mosquito catches primarily by diluting attractants rather than by exceeding mosquito flight capability. Previously published estimates of maximum air speeds of which mosquitoes are capable may now need to be reassessed if they were based upon shut-down of catches in traps baited with chemical attractants. Addition of DEET vapor significantly reduced the number of mosquitoes caught relative to equivalently moving air without repellent. We recommend that fan-generated wind should be pursued as a practical means of protecting humans or pets from mosquitoes in the backyard setting.

Reduction of mosquito (Diptera: Culicidae) attacks on a human subject by combination of wind and vapor-phase DEET repellent.

In a Central Michigan wetland setting, air drawn through a DEET-impregnated screen using an electric fan and projected toward a human subject significantly reduced mosquito orientation by 74%, landing by 75%, and probing by 70%, relative to no applied wind or DEET. The DEET vapor effect was significant as revealed by a statistically significant wind/DEET interaction. The wind speed at the downwind human subject was 0.6 m/s and the DEET vapor concentration was estimated at 2 microg/liter air. We suggest a combination of directed wind and volatile repellent might be developed as a mosquito deterrent strategy for the backyard setting.