Structural basis of impaired disaggregase function in the oxidation-sensitive SKD3 mutant causing 3-methylglutaconic aciduria.

Mitochondria are critical to cellular and organismal health. To prevent damage, mitochondria have evolved protein quality control machines to survey and maintain the mitochondrial proteome. SKD3, also known as CLPB, is a ring-forming, ATP-fueled protein disaggregase essential for preserving mitochondrial integrity and structure. SKD3 deficiency causes 3-methylglutaconic aciduria type VII (MGCA7) and early death in infants, while mutations in the ATPase domain impair protein disaggregation with the observed loss-of-function correlating with disease severity. How mutations in the non-catalytic N-domain cause disease is unknown. Here, we show that the disease-associated N-domain mutation, Y272C, forms an intramolecular disulfide bond with Cys267 and severely impairs SKD3Y272C function under oxidizing conditions and in living cells. While Cys267 and Tyr272 are found in all SKD3 isoforms, isoform-1 features an additional α-helix that may compete with substrate-binding as suggested by crystal structure analyses and in silico modeling, underscoring the importance of the N-domain to SKD3 function.

Laser frequency stabilization in the 10 -14 range via optimized modulation transfer spectroscopy on the 87Rb D2 line.

We present a high-performance laser frequency stabilization method using modulation transfer spectroscopy (MTS) on the rubidium 87D2 transition line. A substantial improvement of the laser frequency stability was achieved by searching for the optimal diameter and intensity settings of the probe and pump beam. The frequency instability measured from the beat frequency of two locked external cavity diode lasers (ECDLs) reached a short-term stability of 4.5×10-14/τ and did not exceed 2 × 10-12 until 105 s, which is the best performance reported thus far with a D2 transition. The long-term stability is limited by the offset fluctuations of the baseline induced by the residual amplitude modulation (RAM), which can be further improved by reducing the current temperature variation of about 0.2 K by means of temperature stabilization or through a further reduction of the RAM.

Deciphering the mechanism and function of Hsp100 unfoldases from protein structure.

Hsp100 chaperones, also known as Clp proteins, constitute a family of ring-forming ATPases that differ in 3D structure and cellular function from other stress-inducible molecular chaperones. While the vast majority of ATP-dependent molecular chaperones promote the folding of either the nascent chain or a newly imported polypeptide to reach its native conformation, Hsp100 chaperones harness metabolic energy to perform the reverse and facilitate the unfolding of a misfolded polypeptide or protein aggregate. It is now known that inside cells and organelles, different Hsp100 members are involved in rescuing stress-damaged proteins from a previously aggregated state or in recycling polypeptides marked for degradation. Protein degradation is mediated by a barrel-shaped peptidase that physically associates with the Hsp100 hexamer to form a two-component system. Notable examples include the ClpA:ClpP (ClpAP) and ClpX:ClpP (ClpXP) proteases that resemble the ring-forming FtsH and Lon proteases, which unlike ClpAP and ClpXP, feature the ATP-binding and proteolytic domains in a single polypeptide chain. Recent advances in electron cryomicroscopy (cryoEM) together with single-molecule biophysical studies have now provided new mechanistic insight into the structure and function of this remarkable group of macromolecular machines.

Moving-frame imaging of transiting cold atoms for precise long-range transport.

Transporting cold atoms between interconnected vacuum chambers is an important technique for increasing the versatility of cold atom setups, particularly for those that couple atoms to photonic devices. In this report, we introduce a method where we are able to image the atoms at all points during transport via moving optical dipole trap. Cooled 87Rb atoms are transported ∼50 cm into an auxiliary vacuum chamber while being monitored with a moving-frame imaging system for which in-situ characterization of the atom transport is demonstrated. Precise positioning of the atoms near photonic devices is also tested across several tapered fibers showing an axial positioning resolution of ∼450 µm.

Maximized atom number for a grating magneto-optical trap via machine-learning assisted parameter optimization.

We present a parameter set for obtaining the maximum number of atoms in a grating magneto-optical trap (gMOT) by employing a machine learning algorithm. In the multi-dimensional parameter space, which imposes a challenge for global optimization, the atom number is efficiently modeled via Bayesian optimization with the evaluation of the trap performance given by a Monte-Carlo simulation. Modeling gMOTs for six representative atomic species - 7Li, 23Na, 87Rb, 88Sr, 133Cs, 174Yb - allows us to discover that the optimal grating reflectivity is consistently higher than a simple estimation based on balanced optical molasses. Our algorithm also yields the optimal diffraction angle which is independent of the beam waist. The validity of the optimal parameter set for the case of 87Rb is experimentally verified using a set of grating chips with different reflectivities and diffraction angles.

Chip-Scale Ultra-Low Field Atomic Magnetometer Based on Coherent Population Trapping.

We report a chip-scale atomic magnetometer based on coherent population trapping, which can operate near zero magnetic field. By exploiting the asymmetric population among magnetic sublevels in the hyperfine ground state of cesium, we observe that the resonance signal acquires sensitivity to magnetic field in spite of degeneracy. A dispersive signal for magnetic field discrimination is obtained near-zero-field as well as for finite fields (tens of micro-tesla) in a chip-scale device of 0.94 cm3 volume. This shows that it can be readily used in low magnetic field environments, which have been inaccessible so far in miniaturized atomic magnetometers based on coherent population trapping. The measured noise floor of 300 pT/Hz1/2 at the zero-field condition is comparable to that of the conventional finite-field measurement obtained under the same conditions. This work suggests a way to implement integrated atomic magnetometers with a wide operating range.

Administration of encapsulated L-tryptophan improves duodenal starch digestion and increases gastrointestinal hormones secretions in beef cattle.

OBJECTIVE: This study investigated the effects of oral administration of rumen-protected L-tryptophan (RPL-T) on duodenal starch digestion and gastrointestinal hormones (GIH) secretions using Hanwoo beef steers as the animal models. METHODS: Four steers (423±24 kg) fitted with ruminal and duodenal cannulas were employed in a crossover design replicated twice. Treatments were control (basal diet) and RPL-T (basal diet+191.1 mg/kg body weight [BW]) group. Blood and duodenal samples were collected to measure serum GIH levels and pancreatic α-amylase activity at day 0, 1, 3, and 5 (-30, 30, 90, 150, and 210 min) of the study. Samples from each segment of the gastrointestinal tract were collected via ruminal and duodenal cannulas and were used to determine soluble protein and the starch digestion rate at days 6 (-30, 180, 360, and 540 min) and 8 (-30, 90, 270, and 450 min) of the experiment. RESULTS: No significant difference in ruminal pH, NH3-N, and total volatile fatty acid including the levels of acetate, propionate, butyrate, isobutyrate, valerate, isovalerate, and the acetate-to-propionate ratio was observed between groups (p>0.05). Crude protein uptake was higher and feces starch content was lower in RPL-T group than the control group (p<0.05). The D-glucose contents of feces in RPL-T group decreased at day 5 compared to those in the control group (p<0.05), however, no change was found at day 0, 1, or 3 compared to the control group (p>0.05). Serum cholecystokinin (CCK), melatonin, duodenal pancreatic α-amylase activity, and starch digestion were significantly higher in RPL-T group than the control group (p<0.05). CONCLUSION: Taken together, oral administration of RPL-T at the rate of 191.1 mg/kg BW consistently increased CCK concentration, pancreatic α-amylase activity in duodenal fluids, and starch digestion rate in the small intestine and thus found to be beneficial.

Intravenous administration of L-tryptophan stimulates gastrointestinal hormones and melatonin secretions: study on beef cattle.

This study aimed to determine the effective dose of intravenous administration of L-tryptophan (L-T) on gastrointestinal hormones (GIH) secretions and melatonin using Hanwoo cattle. Three steers (362 ± 23 kg) fitted with indwelling jugular vein catheters were assigned in a 3 × 3 Latin square design. Treatments were intravenous administration of saline (control), 28.9 mg L-T/kg body weight (BW; low) and 57.8 mg L-T/kg BW (high) L-T for 1 day with 7 days of adaptation. Samples were collected after adaptation period at -60, 0, 30, 60, 90, 120, 150, 180, 240, and 300 min of sampling day. The levels of serum cholecystokinin (CCK) and secretin were higher (p < 0.05) in the high L-T group than those in the other groups. Serum Melatonin (MEL) levels were increased upon L-T administration (p < 0.05) in the high L-T group. Taken together, the effective dose of L-T administration was defined at 57.8 mg L-T/kg BW in order to stimulate increase of GIH and MEL.

Erratum to: Impacts of whey protein on starch digestion in rumen and small intestine of steers.

[This corrects the article DOI: 10.5187/jast.2019.61.2.98.].

Impacts of whey protein on starch digestion in rumen and small intestine of steers.

Four Korean native steers (511 ± 17.2 kg; 2 × 2 replicated crossover design) fitted with duodenal cannulas were used to investigate the influence of oral administration of soluble whey protein (WP; 82.29% crude protein) on ruminal fermentation, gastrointestinal (GI) hormone secretion in the blood, pancreatic α-amylase activity in the duodenum, and disappearance rate in each segment of the GI tract. Steers were orally fed the basal diet (control; TMR [total mixed ration] 9 kg/d) or the basal diet with enriched WP (400 g/d) for 14 days. The apparent crude protein disappearance rate in the rumen of the WP was higher than in control (p < 0.05). However, no difference between groups was observed in the apparent crude protein disappearance rate in the intestine and the apparent starch disappearance rates in the rumen, GI tract. The level of cholecystokinin, secretin, and ghrelin in serum and pancreatic α-amylase activity in the duodenum of the WP also did not change. The changes in the level of blood urea nitrogen related to protein metabolism were higher in the WP than in the control (p < 0.05). However, the levels of total protein, lipid, carbohydrate and mineral metabolites did not change. Consequently, we suggest that the oral administration of WP in steers assisted in ruminal fermentation due to the population increase of microbes in the rumen but did not improve the starch digestion rate in the small intestine because GI hormone secretion in the blood and pancreatic α-amylase activity did not change.

Optical phase-locking of two extended-cavity diode lasers by serrodyne modulation.

We report the optical phase-locking of two extended-cavity diode lasers with a frequency difference of 6.9 GHz by serrodyne modulation. The bandwidth of the phase-locking loop is extended up to 9.5 MHz. The residual phase noise of the two phase-locked lasers reaches -130 dBrad2/Hz in the offset frequency range of 1.5 kHz to 9 kHz and below -120 dBrad2/Hz in the range of 150 Hz to 350 kHz, respectively. It is expected that the sensitivity limit of atom interferometers will be enhanced when the phase-locked lasers are used.

Comparison of Radiographic and Clinical Results After Extended Distal Chevron Osteotomy With Distal Soft Tissue Release With Moderate Versus Severe Hallux Valgus.

BACKGROUND:: We compared the radiographic and clinical outcomes of moderate and severe hallux valgus treated by extended distal chevron osteotomy (EDCO) and distal soft tissue release (DSTR). METHODS:: We performed a retrospective analysis comparing the utility of EDCO for the treatment of moderate (36 patients, 46 feet) and severe (36 patients, 42 feet) hallux valgus. The radiologic outcomes were evaluated based on the preoperative and 3-year follow-up x-rays. The clinical results were investigated based on the 3-year follow-up Manchester-Oxford Foot Questionnaire (MOXFQ), patient satisfaction, and postoperative complications. RESULTS:: The mean postoperative intermetatarsal angle, hallux valgus angle, tibial sesamoid position, and relative metatarsal bone length were significantly different compared with the preoperative values for both the moderate and severe groups ( P < .001). At 3-year follow-up, intergroup differences were observed in the mean postoperative intermetatarsal angle ( P = .001), hallux valgus angle ( P = .003), and tibial sesamoid position ( P = .013); however, mean radiographic results were within the normal range for both groups. There were no intergroup differences for either the first metatarsal bone length ( P = .172) or shortening ( P = .621). No significant difference in MOXFQ ( P = .525) was evident between the groups at 3-year follow-up. In satisfaction analysis, 82.6% of the moderate group reported good to excellent results, as did 81.0% of the severe group ( P = .815). A total of 7 complications were reported in each group ( P = 1.000). CONCLUSION:: The present study showed that midterm mean radiographic results of both moderate and severe hallux valgus treated by EDCO with DSTR were favorable, and those values were within the normal range. Clinical outcomes were comparable between the 2 groups, including overall efficacy. Based on these results, we recommend EDCO with DSTR as an efficient and reliable operative option for both moderate and severe hallux valgus. LEVEL OF EVIDENCE:: Level III, retrospective comparative study.

Differences in youngest-old, middle-old, and oldest-old patients who visit the emergency department.

OBJECTIVE: As aging progresses, clinical characteristics of elderly patients in the emergency department (ED) vary by age. We aimed to study differences among elderly patients in the ED by age group. METHODS: For 2 years, patients aged 65 and older were enrolled in the study and classified into three groups: youngest-old, ages 65 to 74 years; middle-old, 75 to 84 years; and oldest-old, ≥85 years. Participants' sex, reason for ED visit, transfer from another hospital, results of treatment, type of admission, admission department and length of stay were recorded. RESULTS: During the study period, a total 64,287 patients visited the ED; 11,236 (17.5%) were aged 65 and older, of whom 14.4% were 85 and older. With increased age, the female ratio (51.5% vs. 54.9% vs. 69.1%, P<0.001), medical causes (79.5% vs. 81.3% vs. 81.7%, P=0.045), and admission rate (35.3% vs. 42.8% vs. 48.5%, P<0.001) increased. Admissions to internal medicine (57.5% vs. 59.3% vs. 64.7%, P<0.001) and orthopedic surgery (8.5% vs. 11.6% vs. 13.8%, P< 0.001) also increased. The ratio of admission to intensive care unit showed no statistical significance (P=0.545). Patients over age 85 years had longer stays in the ED (330.9 vs. 378.9 vs. 407.2 minutes, P<0.001), were discharged home less (84.4% vs. 78.9% vs. 71.5%, P<0.001), and died more frequently (6.3% vs. 10.4% vs. 13.0%, P<0.001). CONCLUSION: With increased age, the proportion of female patients and medical causes increased. Rates of admission and death increased with age and older patients had longer ED and hospital stays.

Observation of an exceptional point in a two-dimensional ultrasonic cavity of concentric circular shells.

We report observation of an exceptional point in circular shell ultrasonic cavities in both theory and experiment. In our theoretical analysis we first observe two interacting mode groups, fluid- and solid-based modes, in the acoustic cavities and then show the existence of an EP of these mode groups exhibiting a branch-point topological structure of eigenfrequencies around the EP. We then confirm the mode patterns as well as eigenfrequency structure around the EP in experiments employing the schlieren method, thereby demonstrating utility of ultrasound cavities as experimental platform for investigating non-Hermitian physics.

Mie resonance-enhanced pumping and detection efficiency for shallow nitrogen-vacancy centers in diamond.

We investigate Mie resonances of a diamond nano-resonator as a means to enhance the pumping and detection efficiency of shallow nitrogen-vacancy color centers. We show it is possible to tune a couple of high-order modes of a single resonator to each absorption and emission spectrum of the color center, and thereby the resonator plays a dual role of pump field concentration and emission field guiding. Furthermore superposition of the resonator field and the uncoupled near field results in even stronger pump intensity in the shallow top layer of the resonator. We also examine possible coupling between adjacent resonators when they form a periodic array. This approach allows us to achieve lower excitation power and higher signal intensity at local sites defined by resonators providing a way to enhance wide-field metrology in the sampled region of shallow color centers.

Improvement of Milk Fatty Acid Composition for Production of Functional Milk by Dietary Phytoncide Oil Extracted from Discarded Pine Nut Cones (Pinus koraiensis) in Holstein Dairy Cows.

This study was conducted to examine the effects of adding phytoncide oil extracted from Korean pine nut cone byproduct to the diet of dairy cows on milk yield and compositions, fatty acid characteristics, complete blood count and stress response. A total of 74 Holstein cows were used for 30 days and divided into two groups. Each group was given a basal diet (C) or an experimental diet containing phytoncide additives at 0.016% (T) in feed. The results showed that phytoncide feeding had no effect on milk yield. In addition, there were no observed effects on milk composition, but the ratio of fatty acid in milk was significantly affected by the phytoncide diet, and it showed a positive effect. Not only were the major functional fatty acids, conjugated linoleic acid and eicosapentaenoic acid increased, but also ω6:ω3 fatty acid ratio was reduced in milk of T group (p<0.05). In blood analysis, the complete blood count showed no significant difference between C and T group on all parameters. However, the cortisol concentration was significantly decreased in T group compared to control (p<0.05). Taken together, we suggest that phytoncide oil does not have a great influence on the physiological changes, but can be a potential feed additive that improves the milk fatty acid and stress resilience in dairy cows. In addition, it will contribute to the development of feed resource, a reduction in feed cost and a lessening of environmental pollution.

Experimental Observation of Bohr's Nonlinear Fluidic Surface Oscillation.

Niels Bohr in the early stage of his career developed a nonlinear theory of fluidic surface oscillation in order to study surface tension of liquids. His theory includes the nonlinear interaction between multipolar surface oscillation modes, surpassing the linear theory of Rayleigh and Lamb. It predicts a specific normalized magnitude of 0.416η(2) for an octapolar component, nonlinearly induced by a quadrupolar one with a magnitude of η much less than unity. No experimental confirmation on this prediction has been reported. Nonetheless, accurate determination of multipolar components is important as in optical fiber spinning, film blowing and recently in optofluidic microcavities for ray and wave chaos studies and photonics applications. Here, we report experimental verification of his theory. By using optical forward diffraction, we measured the cross-sectional boundary profiles at extreme positions of a surface-oscillating liquid column ejected from a deformed microscopic orifice. We obtained a coefficient of 0.42 ± 0.08 consistently under various experimental conditions. We also measured the resonance mode spectrum of a two-dimensional cavity formed by the cross-sectional segment of the liquid jet. The observed spectra agree well with wave calculations assuming a coefficient of 0.414 ± 0.011. Our measurements establish the first experimental observation of Bohr's hydrodynamic theory.

Killing potential protist predators as a survival strategy of the newly described dinoflagellate Alexandrium pohangense.

Blooms caused by some species belonging to the dinoflagellate genus Alexandrium are known to cause large-scale mortality of fish. Thus, the dynamics of these species is important and of concern to scientists, officials, and people in the aquaculture industry. To understand the dynamics of such species, their growth and mortality due to predation need to be assessed. The newly described dinoflagellate Alexandrium pohangense is known to grow slowly, with a maximum autotrophic growth rate of 0.1d-1. Thus, it may not form bloom patches if its mortality due to predation is high. Therefore, to explore the mortality of A. pohangense due to predation, feeding on this species by the common heterotrophic dinoflagellates Gyrodinium dominans, Gyrodinium moestrupii, Luciella masanensis, Noctiluca scintillans, Oxyrrhis marina, Oblea rotunda, Polykrikos kofoidii, and Pfiesteria piscicida, as well as by the ciliate Tiarina fusus, was examined. None of these potential predators was able to feed on A. pohangense. In contrast, these potential predators were killed and their bodies were dissolved when incubated with A. pohangense cells or cell-free culture filtrates. The survival of G. moestrupii, O. marina, P. kofoidii, and T. fusus on incubation with 10cellsml-1 of A. pohangense was 20-60%, while that at the equivalent culture filtrates was 20-70%. With increasing A. pohangense cell-concentration (up to 1000cellsml-1 or equivalent culture filtrates), the survival rate of G. moestrupii, O. marina, P. kofoidii, and T. fusus rapidly decreased. The lethal concentration (LC50) for G. moestrupii, O. marina, P. kofoidii, and T. fusus at the elapsed time of 24h with A. pohangense cells (cultures of 11.4, 13.3, 1.6, and 3.3cellsml-1, respectively) was lower than that with A. pohangense filtrates (culture filtrates of 35.5, 30.6, 5.5, and 5.0cellsml-1, respectively). Furthermore, most of the ciliates and heterotrophic dinoflagellates in the water collected from the coast of Tongyoung, Korea, were killed when incubated with cultures of 1000 A. pohangense cells ml-1 and equivalent culture filtrates. The relatively slow growing A. pohangense may form blooms by reducing mortality due to predation through killing potential protist predators.

Physiological concentrations of trans-11 18:1 vaccenic acid suppress pro-inflammatory markers under acute inflammation in isolated ICR mice splenocytes.

Direct influences of dietary trans-11 18:1 vaccenic acid (TVA) at physiological concentrations of 50-200 µM were evaluated for cell growth, cytotoxic activity, and cytokine production in leukocytes isolated from the mouse spleen. TVA supplementation for 24 h induced growth of splenocytes at concentrations of 50-200 µM, except for 100 µM. The cytokines TNFα, IFNγ, and IL-10 of splenocytes were stimulated by 100 µM TVA. Induced production of TNFα in splenocytes challenged with lipopolisaccharides was suppressed by 100 µM TVA. Physiological levels of TVA had direct effects on growth and cytokine production in splenocytes. Further in vivo studies are needed to improve understanding of the precise influence of trans fatty acids on production of pro-inflammatory markers under acute inflammation conditions.