Effect of precursors and stress factors on yeast isolated from fermented maesil extract and their biogenic amine formation.

Biogenic amines are produced during fermentation and can act as harmful substances. Strains related to the fermentation of maesil extract were identified and Clavispora lusitaniae and Pichia kluyveri were selected to investigate the relationship between biogenic amines and precursors, NaCl or ethanol. Biogenic amines were analyzed by high-performance liquid chromatography. Among precursors added, arginine was most effective for the biogenic amines formation. After 24 h incubation, the content of total biogenic amines increased from 37.60 to 51.75 mL/L for C. lusitaniae and from 2.60 to 33.30 mL/L for P. kluyveri in arginine-added medium. The number of yeast decreased in both NaCl- and ethanol-YM broth added with arginine, but there was no correlation between the number of yeast and biogenic amines. These results suggested that the formation of biogenic amines by yeast was affected by various factors and their interactions rather than a single factor, such as decarboxylase activity and stress factor. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01328-8.

Effects of Soaking and Fermentation Time on Biogenic Amines Content of Maesil (Prunus Mume) Extract.

Maesil extract, a fruit-juice concentrate derived from Prunus mume prepared by fermenting with sugar, is widely used with increasing popularity in Korea. Biogenic amines in maesil extract were extracted with 0.4 M perchloric acid, derivatized with dansyl chloride, and detected using high-performance liquid chromatography. Among 18 home-made maesil extracts collected from different regions, total biogenic amine content varied from 2.53 to 241.73 mg/L. To elucidate the effects of soaking and fermentation time on biogenic amine content in maesil extract, maesil was soaked in brown sugar for 90 days and the liquid obtained was further fermented for 180 days at 15 and 25 °C, respectively. The main biogenic amines extracted were putrescine and spermidine and the total biogenic amine content was higher at 25 °C than at 15 °C. Soaking at 15 and 25 °C increased the total biogenic amines content from 14.14 to 34.98 mg/L and 37.33 to 69.05 mg/L, respectively, whereas a 180 day fermentation decreased the content from 31.66 to 13.59 mg/L and 116.82 to 57.05 mg/L, respectively. Biogenic amine content was correlated with total amino acid content (particularly, arginine content). Based on these results, we have considered that biogenic amine synthesis can be reduced during maesil extract production by controlling temperature and fermentation time.

Effects of isometric hip movements on electromyographic activities of the trunk muscles during plank exercises.

[Purpose] The purpose of this study was to investigate the effect of isometric hip adduction and abduction on trunk muscle activity during plank exercises. [Subjects and Methods] Nineteen healthy male subjects were recruited for this study. All subjects performed the traditional plank exercise (TP), plank exercise with isometric hip adduction (PHAD), and plank exercise with isometric hip abduction (PHAB) by using an elastic band. Electromyographic (EMG) activities of the internal oblique (IO) and external oblique (EO) were measured during the 3 plank exercises by using an Electromyography system. [Results] Internal oblique and external oblique muscle activities were significantly greater during plank exercise with isometric hip adduction and plank exercise with isometric hip abduction than during traditional plank exercise. Internal oblique and external oblique muscle activities did not differ between the plank exercise with isometric hip adduction and plank exercise with isometric hip abduction conditions. [Conclusion] These findings demonstrate that loaded isometric hip movements may be a useful strategy to increase trunk muscle activity during plank exercises.

Diagnostic performance of abdominal CT for diagnosis of pelvic fractures: comparison with pelvic CT.

BACKGROUND: In the emergency department, patients with abdominopelvic trauma inadvertently undergo both abdominal computed tomography (CT) (for intra-abdominal and intra-pelvic organs) and pelvic CT (CT with multiplanar reformation in three orthogonal planes of the bony pelvis). However, the systemic use of CT is concerning given the cumulative radiation dose. PURPOSE: To evaluate the diagnostic value of abdominal CT in comparison to pelvic CT in patients with suspected pelvic fractures. MATERIAL AND METHODS: Seventy-two patients who underwent abdominal CT and pelvic CT within a 2-week period to evaluate pelvic fractures were included. Two reviewers retrospectively analyzed eight anatomical regions of the pelvic bones on both abdominal CT and pelvic CT over a 1-week interval. The interpretation of pelvic CT scans by two senior musculoskeletal radiologists was considered as the reference standard. Diagnostic performance and inter-observer agreement of both CT scans were evaluated. RESULTS: For reviewers 1 and 2, abdominal CT showed high accuracy (98% and 98%, respectively) as did pelvic CT. For both abdominal CT and pelvic CT, fracture detection in all anatomical regions of the pelvic bones was not significantly different for the two reviewers (P ≥ 0.25). Inter-observer agreement for all anatomical regions of the pelvic bones was excellent or good (k = 0.785-1.0). CONCLUSION: Not only pelvic CT but also abdominal CT is acceptable for detection of pelvic fractures, in spite of its thicker sections and different reconstruction algorithm. Therefore, if abdominal CT has already been performed, additional pelvic CT might no longer be necessary in order to exclude a pelvic fracture.

Increased signal intensity at the proximal patellar tendon: correlation between MR imaging and histology in eight cadavers and clinical MR imaging studies.

OBJECTIVES: We set out to investigate the cause of persistently increased signal intensity (SI) in the posterior portion of the proximal patellar tendon (pPT) on T1-weighted images (T1WI). METHODS: MR imaging was performed in eight cadavers, followed by gross histological examination. In addition, 84 patients without trauma history or anterior knee pain were included to compare the SI of the PTs. The patients were divided according to their age, sex, and Kellgren-Lawrence (KL) grades. The length and thickness of the increased SI portion in the pPT and whole PT (wPT) on T1WI were recorded. RESULTS: Histological specimens demonstrated that the adipose tissue, vessels, and perivascular connective tissue invaginated into the posterior portion of the pPT. This histological anatomy corresponded to the pPT signal change on MR imaging. There was linear and interdigitating increased SI of the pPT in all of the 84 patients (100%). There were no differences in the lengths and thicknesses of the increased SI portion of pPTs and wPTs according to age, sex, and KL grade (all p > 0.05). CONCLUSIONS: The increased SI of the pPT on T1WI and fluid-sensitive MR images results from invaginating fat, vessels, and perivascular connective tissue. It is not pathological, but a normal and common finding. KEY POINTS: • Increased linear/interdigitated SIs of the pPT is a normal and common finding. • Invaginated adipose tissue, vessels, and connective tissue could contribute to increased SI. • The fibrocartilage has a minimal role in increased SI of the pPT. • Age, sex, and KL grade do not significantly influence the increased SI. • Knowledge of this increased SI should help clinicians to avoid unnecessary treatment.

Repaired supraspinatus tendons in clinically improving patients: early postoperative findings and interval changes on MRI.

OBJECTIVE: To demonstrate and further determine the incidences of repaired supraspinatus tendons on early postoperative magnetic resonance imaging (MRI) findings in clinically improving patients and to evaluate interval changes on follow-up MRIs. MATERIALS AND METHODS: Fifty patients, who showed symptomatic and functional improvements after supraspinatus tendon repair surgery and who underwent postoperative MRI twice with a time interval, were included. The first and the second postoperative MRIs were obtained a mean of 4.4 and 11.5 months after surgery, respectively. The signal intensity (SI) patterns of the repaired tendon on T2-weighted images from the first MRI were classified into three types of heterogeneous high SI with fluid-like bright high foci (type I), heterogeneous high SI without fluid-like bright high foci (type II), and heterogeneous or homogeneous low SI (type III). Interval changes in the SI pattern, tendon thickness, and rotator cuff interval thickness between the two postoperative MRIs were evaluated. RESULTS: The SI patterns on the first MRI were type I or II in 45 tendons (90%) and type III in five (10%). SI decreased significantly on the second MRI (p < 0.050). The mean thickness of repaired tendons and rotator cuff intervals also decreased significantly (p < 0.050). CONCLUSION: Repaired supraspinatus tendons exhibited high SI in 90% of clinically improving patients on MRI performed during the early postsurgical period. The increased SI and thickness of the repaired tendon decreased on the later MRI, suggesting a gradual healing process rather than a retear.

Osteochondral lesion of the bilateral femoral heads in a young athletic patient.

Osteochondral lesions of the femoral head are uncommon and few studies have reported their imaging findings. Since joints are at risk of early degeneration after osteochondral damage, timely recognition is important. Osteochondral lesions of femoral head may often be necessary to differentiate from avascular necrosis. Here, we report a case of osteochondral lesions on bilateral femoral heads. This lesion manifested as subchondral cysts in initial radiographs, which led to further evaluation by computed tomography arthrography and magnetic resonance imaging, which revealed overlying cartilage defects.

[A case of cryptococcal meningitis mimicking hepatic encephalopathy in a patient with liver cirrhosis caused by chronic hepatitis C].

Cryptococcus neoformans, an encapsulated fungus, is an important opportunistic pathogen that can cause meningitis in im-munocompromised patients. Since patients with cryptococcemia have high mortality, it is essential to make an early diagnosis and promptly initiate antifungal therapy. However, it is often very difficult to differentiate between cryptococcal meningitis and hepatic encephalopathy in patients with liver cirrhosis, and there is delay in making the diagnosis. Therefore, these patients have a particularly grave prognosis and consequently many patients die before culture results become available. In one study, starting antifungal therapy within 48 hours of the blood culture was associated with improved survival, but patients with liver cirrhosis were significantly less likely to receive antifungal therapy within 48 hours compared to those without liver cirrhosis. Recently, the authors experience a case of a 68-year-old woman with liver cirrhosis who presented with fever and a drowsy mental status. She had a previous history of having been admitted for infection-associated hepatic encephlopathy. Cryptococcal meningitis and cryptococcemia were diagnosed by spinal puncture and culture of cerebrospinal fluid. In spite of adequate treatment, the patient developed multi-system organ failure and eventually expired. Herein, we report a case of cryptococcal meningitis mimicking hepatic encephalopathy in a patient with liver cirrhosis.

Changes in dissociation efficiency and kinetics of peptide ions induced by basic residues and their mechanistic implication.

With matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry, total abundance of product ions formed by dissociation inside (in-source decay, ISD) and outside (post-source decay, PSD) the source was measured for peptide ions [Y(5)X + H](+), [XY(5) + H](+), [Y 2)XY(3) + H](+), and [XY(4)X + H](+) (X = tyrosine (Y), histidine (H), lysine (K), and arginine (R) with H for the ionizing proton). α-Cyano-4-hydroxycinammic acid was used as matrix. Product abundance became smaller in the presence of basic residues (H, K, and R), in the order Y > H ≈ K > R. In particular, product abundances in ISD of peptide ions with R were smaller than those with H or K by an order of magnitude, which, in turn, were smaller than that for [Y(6) + H](+) by an order of magnitude. Product abundance was affected by the most basic residue when more than one basic residue was present. A kinetic explanation for the data was attempted under the assumption of quasi-thermal equilibrium for peptide ions in MALDI plume which undergoes expansion cooling. Dramatic disparity in product abundance was found to arise from small difference in critical energy and entropy. Results indicate similar transition structures regardless of basic residues present, where the ionizing proton keeps interacting with a basic site. Further implication of the results on the dissociation mechanism along b-y channels is discussed.

A comparative study of in- and post-source decays of peptide and preformed ions in matrix-assisted laser desorption ionization time-of-flight mass spectrometry: effective temperature and matrix effect.

In-source decay (ISD) and post-source decay (PSD) of a peptide ion ([Y(6) + H](+)) and a preformed ion (benzyltriphenylphosphonium, BTPP) generated by matrix-assisted laser desorption ionization (MALDI) were investigated with time-of-flight mass spectrometry. α-Cyano-4-hydroxycinammic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB) were used as matrices. For both ions, ISD yield was unaffected by delay time, indicating rapid termination of ISD. This was taken as evidence for rapid expansion cooling of hot "early" plume formed in MALDI. CHCA was hotter than DHB for [Y(6) + H](+) while the matrix effect was insignificant for BTPP. The "early" plume temperature estimated utilizing previous kinetic results was 800-900 K, versus 400-500 K for "late" plume. The results support our previous finding that the temperature of peptide ions interrogated by tandem mass spectrometry was lower than most rough estimates of MALDI temperature.

Dissociation mechanisms and implication for the presence of multiple conformations for peptide ions with arginine at the C-terminus: time-resolved photodissociation study.

Time-resolved photodissociation (PD) patterns of singly protonated peptides with arginine at the C-terminus (C-arg peptide ions) have been used to classify the dissociation channels into two categories, i.e. high-energy channels generating v, w and x and low-energy ones generating b, y and z. x + 1 formed by C(alpha)-CO cleavage seems to be the intermediate ion in high-energy channels just as a + 1 is for N-arg peptide ions. Difference in time-resolved pattern indicates that the two sets of channels, high- and low-energy ones, are not in direct competition. Noncompetitive dissociation is also indicated by the observation of anomalous effect of matrix used in matrix-assisted laser desorption ionization, a cooler matrix generating more high-energy product ions both in spontaneous dissociation and in PD. Results from detailed investigation suggest that the two sets of channels start from two (or more) different conformations.

Dissociation kinetics of singly protonated leucine enkephalin investigated by time-resolved photodissociation tandem mass spectrometry.

The yields of post-source decay (PSD) and time-resolved photodissociation (PD) at 193 and 266 nm were measured for singly protonated leucine enkephalin ([YGGFL + H](+)), a benchmark in the study of peptide ion dissociation, by using tandem time-of-flight mass spectrometry. The peptide ion was generated by matrix-assisted laser desorption ionization (MALDI) using 2,5-dihydroxybenzoic acid as the matrix. The critical energy (E(0)) and entropy (DeltaS(++) at 1000 K) for the dissociation were determined by Rice-Ramsperger-Kassel-Marcus fit of the experimental data. MALDI was done for a mixture of YGGFL and Y(6) and the plume temperature determined by the kinetic analysis of [Y(6) + H](+) data were used to improve the precision of E(0) and DeltaS(++) for [YGGFL + H](+). E(0) and DeltaS(++) thus determined (E(0) = 0.67 +/- 0.08 eV, DeltaS(++) = -24.4 +/- 3.2 eu with 1 eu = 4.184 J K(-1)mol(-1)) were significantly different from those determined by blackbody infrared radiative dissociation (BIRD) (E(0) = 1.10 eV, DeltaS(++) = -14.9 eu), and by surface-induced dissociation (SID) (E(0) = 1.13 eV, DeltaS(double dagger) = -10.3 eu). Analysis of the present experimental data with the SID kinetics (and BIRD kinetics also) led to an unrealistic situation where not only PSD and PD but also MALDI-TOF signals could not be detected. As an explanation for the discrepancy, it was suggested that transition-state switching occurs from an energy bottleneck (SID/BIRD) to an entropy bottleneck (PSD/PD) as the internal energy increases.

Influence of basic residues on dissociation kinetics and dynamics of singly protonated peptides: time-resolved photodissociation study.

Product ion yields in postsource decay and time-resolved photodissociation at 193 and 266 nm were measured for some peptide ions with lysine ([KF6 + H]+, [F6K + H]+, and [F3KF3 + H]+) formed by matrix-assisted laser desorption ionization. The critical energy (E0) and entropy (DeltaS(double dagger)) were determined by RRKM fitting of the data. The results were similar to those found previously for peptide ions with histidine. To summarize, the presence of a basic residue, histidine or lysine, inside a peptide ion retarded its dissociation by lowering DeltaS(double dagger). On the basis of highly negative DeltaS(double dagger), presence of intramolecular interaction involving a basic group in the transition structure was proposed.

Time-resolved photodissociation study of singly protonated peptides with a histidine residue generated by matrix-assisted laser desorption ionization: dissociation rate constant and internal temperature.

Product ion yields in post-source decay and time-resolved photodissociation at 193 and 266 nm were measured for some peptide ions with a histidine residue ([HF(6) + H](+), [F(6)H + H](+), and [F(3)HF(3) + H](+)) formed by matrix-assisted laser desorption ionization (MALDI). Compared with similar data for peptide ions without any basic residue reported previously, significant reduction in dissociation efficiency was observed. Internal temperatures (T) of the peptide ions and their dissociation kinetic parameters-the critical energy (E(0)) and entropy (DeltaS(double dagger))-were determined by the method reported previously. Slight decreases in E(0), DeltaS(double dagger), and T were responsible for the histidine effect-reduction in dissociation rate constant. Regardless of the presence of the residue, DeltaS(double dagger) was far more negative than previous quantum chemical results. Based on this, we propose the existence of transition structures in which the nitrogen atoms in the histidine residue or at the N-terminus coordinate to the reaction centers. Reduction in T in the presence of a histidine residue could not be explained based on popular models for ion formation in MALDI, such as the gas-phase proton transfer model.

Temperature of peptide ions generated by matrix-assisted laser desorption ionization and their dissociation kinetic parameters.

Product ion yields in postsource decay and photodissociation at 193 and 266 nm were measured for some peptide ions without a basic amino acid residue ([Y(6) + H](+), [F(5) + H](+), and [YPFVEPI + H](+)) generated by matrix-assisted laser desorption ionization (MALDI). Data indicated statistical nature for the dissociation processes. Assuming that peptide ions formed by MALDI are in thermal equilibrium at temperature T and that their dissociation rate constants are specified by the critical energy (E(0)) and entropy (DeltaS(double dagger)), a method based on kinetic analysis was devised to determine these parameters simultaneously. The matrix used was found to affect the effective temperature of peptide ions, 2,5-dihydroxybenzoic acid (400-430 K) < sinapinic acid (440 K) < alpha-cyano-4-hydroxycinnamic acid (460-510 K), in agreement with previous perceptions. E(0) of around 0.6 eV and DeltaS(double dagger) of -24 eu were smaller than previous quantum chemical results for small model peptide ions.

Time-resolved photodissociation of singly protonated peptides with an arginine at the N-terminus: a statistical interpretation.

Time-evolution of product ion signals in ultraviolet photodissociation (UV-PD) of singly protonated peptides with an arginine at the N-terminus was investigated by using a tandem time-of-flight mass spectrometer equipped with a cell floated at high voltage. Observation of different time-evolution patterns for different product ion types--an apparently nonstatistical behavior--could be explained within the statistical framework by invoking consecutive formation of some product ions and broad internal energy distributions for precursor ions. a(n) + 1 and b(n) ions were taken as the primary product ions from this type of peptide ions. Spectral characteristics in post-source decay, UV-PD, and collisionally activated dissociation at low and high kinetic energies could be explained via rough statistical calculation of rate constants. Specifically, the striking characteristics in high-energy CAD and UV-PD--dominance of a(n) and d(n) formed via a(n) + 1--were not due to the peculiarity of the excitation processes themselves, but due to quenching of the b(n) channels caused by the presence of arginine.

Development of a time-resolved method for photodissociation mechanistic study of protonated peptides: use of a voltage-floated cell in a tandem time-of-flight mass spectrometer.

Photodissociation at 266 nm of some protonated peptides was investigated using a tandem-TOF spectrometer equipped with a cell near its first time focal point where the laser was irradiated. When a high voltage was applied to the cell, each product ion peak split into several components with different flight times. One of these was due to in-cell direct formation of the product ion and another due to post-cell formation. Those in between were due to consecutive dissociations, the first steps of which occurred inside the cell and the second steps outside the cell. A method based on flight time calculation was developed to analyze these components and to identify the intermediate ion for each consecutive component. The technique allows time-resolved photodissociation mechanistic studies on a 100-ns timescale.

Characteristics of photodissociation at 193 nm of singly protonated peptides generated by matrix-assisted laser desorption ionization (MALDI).

Photodissociation (PD) at 193 nm of various singly protonated peptides was investigated. These include peptides with an arginine residue at the C-terminus, N-terminus, at both termini, inside the chain, and those without an arginine residue. Monoisotopomeric selection was made for the precursor ions. Interference from the post-source decay (PSD) product signals was reduced as much as possible by using the deflection system (reported previously) and subtracting the remaining signals from the laser-on signals. The presence of an arginine residue and its position inside the peptide were found to significantly affect the PD spectra, as reported previously. Presence of a proline, aspartic acid, or glutamic acid residue hardly affected the PD spectral patterns. By comparing the PD spectra obtained at a few different wavelengths, it is concluded that the dissociation of the photoexcited ions occurs in their ground electronic states. Tentative explanations for the observed spectral correlations based on the statistical picture for the reactions are also presented.

A deflection system to reduce the interference from post-source decay product ions in photodissociation tandem time-of-flight mass spectrometry.

A deflection system consisting of four deflectors was designed and used to reduce the interference from post-source decay (PSD) product ions in photodissociation (PD) tandem time-of-flight (TOF) mass spectrometry. For simple protonated peptides generated by matrix-assisted laser desorption/ionization, the presence of PSD product ions at the laser irradiation spot was found to noticeably alter the minor peaks in the PD spectra even though the major ones were hardly affected. Other benefits from the use of the deflection system such as the improvement in the resolving power in PSD tandem mass spectra are discussed.

Photodissociation of singly protonated peptides at 193 nm investigated with tandem time-of-flight mass spectrometry.

Photodissociation at 193 nm of some singly protonated peptides generated by matrix-assisted laser desorption/ionization was investigated using tandem time-of-flight mass spectrometry. For peptides with arginine at the C-terminus, x, upsilon, and w fragment ions were generated preferentially while a and d fragment ions dominated for peptides with arginine at the N-terminus. These are the same characteristics as photodissociation at 157 nm reported previously. Overall, the photodissociation spectra obtained at 157 and 193 nm were strikingly similar.