The extent of unnecessary tooth loss due to extractions prior to radiotherapy based on radiation field and dose in patients with head and neck cancer.
Prior to radiotherapy (RT), teeth with poor prognosis that pose a risk for post-RT osteoradionecrosis (ORN) are removed. To allow enough time for adequate wound healing prior to RT, decisions are made based on the estimated radiation dose. This study aimed to gain insight into (1) the overall number of teeth extracted and (2) the patient and tumor characteristics associated with the number of redundantly extracted teeth. Patients with head and neck cancer (HNC), treated with RT between 2015 and 2019, were included in this cross-sectional study. For each extracted tooth the radiation dose was calculated retrospectively. The cut-off point for valid extraction was set at ≥ 40 Gy in accordance with the national protocol. Potential factors for doses ≥ 40 Gy were identified, including age, sex, tumor location, tumor (T) and nodal stage (N), overall tumor stage and number of teeth extracted. A total of 1759 teeth were removed from 358 patients. Of these 1759 teeth, 1274 (74%) appeared to have been removed redundantly, based on the mean dose (Dmean) of < 40 Gy. Using the maximum dose (Dmax) of < 40 Gy, 1080 teeth (61%) appeared to have been removed redundantly. Tumor location and N-classification emerged as the most important associative variables in the multivariable regression analysis. To our knowledge this is the first study to provide insight into the amount of teeth redundantly extracted prior to RT and represents a step forward in de-escalating the damage to the masticatory system prior to RT.


Laboratory life cycle of Ornithodoros puertoricensis (Ixodida: Argasidae) collected in the Colombian Caribbean.
Three studies on the biology of Ornithodoros puertoricensis are available in the literature, using different hosts and incubation temperatures. In a previous study, we identified O. puertoricensis in the Colombian Caribbean. The aim of the present work was to analyze life cycle data along one generation from these specimens under laboratory conditions. Eggs of O. puertoricensis were collected in between fragments of bahareque material in a rural dwelling in the municipality of Planeta Rica (Córdoba Department, Colombia), and transported to the laboratory. All post-egg stages (i.e., larvae, nymphs, and adults) were incubated at 27 °C and 85% RH and fed on laboratory mice (Mus musculus). Sixteen engorged larvae were obtained to start a laboratory colony. Average feeding period for larvae was 4.6 days (4-5). The first nymphal instar (N1) did not require feeding and the subsequent nymphal stages (N2, N3, and N4) and adults had feeding periods ranging from 55 to 75 min. Average pre-molting period in nymphs was 15 days (10-21). Most of the N3 molted to males and all N4 molted to females. Two gonotrophic cycles were recorded: the first had a preoviposition period of 12 days (7-18) and produced 190 eggs (171-223), the second lasted 6.6 days (6-7) and produced 146 eggs (104-201). The mean life cycle duration (from parental eggs to F1 eggs) was 70.7 days (58.7-82.7) without fasting periods. The collected data agree with previous studies even with differences in hosts and maintenance conditions.


A prospective randomized controlled trial evaluating alcohol on loudness perception in cochlear implant users.
The aim of this study was to determine the effects of alcohol on the psychophysical responses in patients with cochlear implants. This has not been previously studied. It was also hoped to provide information that could suggest possible sites of action of the known effects of alcohol on the auditory pathway. A prospective randomized placebo-controlled trial, with full ethical approval. Eight successful cochlear implant users were selected, of whom two had bilateral implants which were tested separately. In total 10 cochlear implants were tested. Alcohol was given in the form of vodka (50% alcohol, 1 mL/kg body weight) with 500 mL of orange and cranberry juice. The placebo control was given in the form of 500 mL of orange and cranberry juice alone. The 'comfort level' (C level) was recorded before, and 1 h after alcohol or placebo ingestion for each patient's cochlear implant. Blood alcohol concentration was determined prior to alcohol or placebo consumption and then repeated after 45, 60, 90 and 180 min. The mean blood alcohol concentration 1 h after ingestion was 50 mg/dL. In the 'alcohol' arm the mean electrical unit increase in the C level was 19.9 with a standard deviation of 2.2. In the control arm the mean change in C level was 0.10 with a standard deviation of 0.3. In this first prospective randomized control study of the effect of alcohol on sound perception in cochlear implant users, alcohol significantly increased the upper end of the dynamic range (C levels) in comparison with placebo (P = <0.0001 using paired t-test analysis). This effect is likely to be the result of change in the auditory pathways proximal to the cochlea.


Perception that oral anticancer treatments are less efficacious: development of a questionnaire to assess the possible prejudices of patients with cancer.
We developed and pilot tested a 12-item questionnaire, to approach the issue of patients' perception on efficacy of oral chemotherapy. An additional question was on 'trade-off' between treatment efficacy and the ease of oral administration. The motivating underlying hypothesis was that oral drugs might be perceived by patients as less effective than when drugs were delivered by injection. The questionnaire was given to 59 patients with advanced breast cancer who received oral chemotherapy. Ninety-percent of patients considered clear and completed the questionnaire. Oral chemotherapy was positively viewed by most patients, perceiving it as advantageous (58%), able to help them feel less ill (77%) and to reduce the effort in coping with the disease (67%). The most important feeling elicited was the sense of freedom. Younger patients (<45 years) required bigger effort to face their illness and expected the most from oral chemotherapy. The questionnaire was shown to be robust, reliable and reproducible instrument in assessing patients' prejudices towards oral anticancer treatments.


Inter-individual variations of human mercury exposure biomarkers: a cross-sectional assessment.
Biomarkers for mercury (Hg) exposure have frequently been used to assess exposure and risk in various groups of the general population. We have evaluated the most frequently used biomarkers and the physiology on which they are based, to explore the inter-individual variations and their suitability for exposure assessment. Concentrations of total Hg (THg), inorganic Hg (IHg) and organic Hg (OHg, assumed to be methylmercury; MeHg) were determined in whole blood, red blood cells, plasma, hair and urine from Swedish men and women. An automated multiple injection cold vapour atomic fluorescence spectrophotometry analytical system for Hg analysis was developed, which provided high sensitivity, accuracy, and precision. The distribution of the various mercury forms in the different biological media was explored. About 90% of the mercury found in the red blood cells was in the form of MeHg with small inter-individual variations, and part of the IHg found in the red blood cells could be attributed to demethylated MeHg. THg in plasma was associated with both IHg and MeHg, with large inter-individual variations in the distribution between red blood cells and plasma. THg in hair reflects MeHg exposure at all exposure levels, and not IHg exposure. The small fraction of IHg in hair is most probably emanating from demethylated MeHg. The inter-individual variation in the blood to hair ratio was very large. The variability seemed to decrease with increasing OHg in blood, most probably due to more frequent fish consumption and thereby blood concentrations approaching steady state. THg in urine reflected IHg exposure, also at very low IHg exposure levels. The use of THg concentration in whole blood as a proxy for MeHg exposure will give rise to an overestimation of the MeHg exposure depending on the degree of IHg exposure, why speciation of mercury forms is needed. THg in RBC and hair are suitable proxies for MeHg exposure. Using THg concentration in plasma as a measure of IHg exposure can lead to significant exposure misclassification. THg in urine is a suitable proxy for IHg exposure.


Tissue-specific genotoxicity and antioxidant imbalance of titanium dioxide nanoparticles (NPTiO2) and inorganic lead (PbII) in a neotropical fish species.
The aquatic environment is the major recipient of wastes containing nanoparticles and other contaminants. Titanium dioxide nanoparticles (NPTiO2) are one of the most produced and used nanoparticle worldwide. This study investigated the toxicity of NPTiO2, as well as the toxicity interaction between NPTiO2 and lead (Pb), in response to genetic and biochemical biomarkers using freshwater fish Rhamdia quelen, as an animal model. The results showed genotoxicity in blood and kidney tissues. No effect of NPTiO2 alone or in co-exposure with Pb on liver genotoxicity were observed. Alterations in the antioxidant hepatic enzymes activities, as well as alterations in glutathione levels indicated that NPTiO2 alone or in co-exposure with Pb can cause antioxidant imbalance. The lipid peroxidation was also raised after exposure to NPTiO2. In general, the results of this study indicated that both NPTiO2 alone and their co-exposure with Pb are capable of producing significant toxic effects in short-term exposure.


Coupled plasmon-phonon modes in monolayer MoS2.
We present a theoretical study of the plasmon-phonon coupling in a suspended monolayer MoS2 and a MoS2 substrate system using a diagrammatic self-consistent field theory. The four coupled plasmon-phonon modes and the four plasmon-surface phonon modes are observed due to the spin-orbit and electron-optic phonon interactions. The two of coupled plasmon-phonon and plasmon-surface phonon modes are optic-like and the other two are acoustic-like. The plasmon are strongly coupled with the optic-phonon in MoS2 and the surface optic-phonon in the substrates as the electron density or wave-vector increases. The strong plasmon-phonon coupling shows that the optoelectronic properties of monolayer MoS2 are evidently modulated by electron-phonon interactions. The hybrid plasmon-phonon polaritons can be achieved by strong light-mater interactions. This study is relevant to the application of MoS2 as novel plasmonic and nanophotonic devices.


Amphiphilic (di-)gradient copoly(2-oxazoline)s are potent amyloid fibril formation inhibitors.
Amyloidoses are diseases caused by the accumulation of normally soluble proteins in the form of insoluble amyloids, leading to the gradual dysfunction and failure of various organs and tissues. Inhibiting amyloid formation is therefore an important therapeutic target. We hypothesized that mono- and di-gradient amphiphilic copolymers of hydrophilic 2-(m)ethyl-2-oxazoline and hydrophobic 2-aryl-2-oxazolines may inhibit amyloid fibril formation. In the model system with hen egg white lysozyme (HEWL) as amyloidogenic protein we determined the effect of these polymers on the amyloid formation by making use of the thioflavin T fluorescence, transmission electron microscopy, isothermal titration calorimetry, and dynamic light scattering. We found that some gradient copolymers possess very potent concentration-dependent inhibitory effects on HEWL amyloid formation. Structure-activity relationship revealed that copolymers with higher ratios of aromatic monomeric units had stronger amyloid suppression effects, most plausibly due to the combination of hydrophobic and π-π interactions. The measurements also revealed that the polymers that inhibit amyloid formation most plausibly do so in the form of micelles that interact with the growing amyloid fibril ends, not with isolated HEWL molecules in solution. These findings suggest the potential use of these gradient copolymers as therapeutic agents for amyloidoses.


The variation of PM2.5 from ship emission under low-sulfur regulation: A case study in the coastal suburbs of Kitakyushu, Japan.
From January 1, 2020, the International Maritime Organization (IMO) regulation about the limit of fuel sulfur content to 0.5 % become effective, and ships commonly install sulfur scrubbers or use low-sulfur fuel or liquefied natural gas to replace sulfur-rich heavy fuel oil. In this study, the 4-year PM2.5 sampling in the coastal suburbs of Kitakyushu, Japan clearly indicated the significant effects of relevant regulation and countermeasures on particle emissions in this receptor site. From the perspective of air quality, an obvious decrease in the mass concentration of ship-emitted particles was observed in 2020, and the contribution of sulfate could reach 60 %. The ammonium concentration was mainly controlled by sulfate and nitrate, and its reduction also could not be ignored, accounting for about 17 %. In terms of public health, the particle exposure risk also changed greatly, mainly due to the reduction of risk levels for As, W, Sb, V, Ni, and Cd; the lowest non-carcinogenic risk and carcinogenic risk for both adults (HI = 1.2 and CR = 5.7 × 10-5) and children (HI = 9.9 and CR = 1.1 × 10-4) all occurred in 2020. However, these reduced health risks were still not within the safe level (except for the carcinogenic risk for adults), a fact that requires continued attention. This result exposed the deficiency of current countermeasures regarding the IMO's fuel sulfur content limit in Kitakyushu City, and increasing the proportion of ships using clean fuels (liquefied natural gas, methanol, etc.) would surely alleviate the particle pollution caused by ship emissions.


Prospective study of a Trucut needle for performing EUS-guided biopsy with EUS-guided FNA rescue.
EUS-guided FNA (EUS-FNA) is an accurate technique for sampling extraintestinal masses and lymph nodes. The use of a Trucut needle to perform EUS-guided biopsy (EUS-TCB) may improve the results or simplify the procedure. To date, few studies have prospectively assessed the performance and the safety of EUS-TCB. Patients with a known or a suspected malignancy referred for a diagnostic and/or staging EUS examination were enrolled in a prospective study. EUS-guided biopsy was performed first with a 19-gauge Trucut needle. If the Trucut failed to obtain an adequate sample or when the "in room" touch preparation was benign, EUS-FNA was performed with a standard 22-gauge FNA needle. The objective of the study was to assess the yield of detection of malignancy and the safety of EUS-TCB in patients with known or suspected malignancies and to investigate if EUS-FNA has a role for rescue in cases of Trucut failure. Thirty-nine lesions underwent EUS-TCB in 30 patients. Sufficient follow-up was available for all patients. By using EUS-TCB, we were able to obtain a sample for diagnosis in all but 3 patients (one pancreatic mass and two lymph nodes) in which technical problems arose. In these patients, the diagnosis was obtained in two cases by EUS-FNA and in the other one by EUS-TCB from the primary pancreatic tumor. The yield of detection of malignancy for EUS-TCB was 84%. No complications were recorded in any patients at 1 and 7 days of follow-up. The sample size is limited to generalize conclusions. EUS-TCB is a safe and an accurate procedure to obtain a histologic diagnosis in patients with known or suspected malignancies. EUS-FNA can serve as a rescue technique in cases of Trucut failure.


Strain induced metal-semiconductor transition in two-dimensional topological half metals.
Spintronic applications of two-dimensional (2D) magnetic half metals and semiconductors are thought to be very promising. Here, we suggest a family of stable 2D materials M n 2 X 7 (X = Cl, Br, and I). The monolayer M n 2 C l 7 exhibits an in-plane ferromagnetic (FM) ground state with a Curie temperature of 118 K, which is unveiled to be a 2D Weyl half semimetal with two Weyl points of opposite chirality connected by a remarkable Fermi arc. In addition, it appears that a biaxial tensile strain can lead to a metal-semiconductor phase transition as a result of the increased anomalous Jahn-Teller distortions, which raise the degeneracy of the e g energy level and cause a significant energy splitting. A 10% biaxial tensile strain also increases the Curie temperature to about 159 K, which originates from the enhanced Mn-Cl-Mn FM superexchange. Moreover, the metal-semiconductor transition can also be induced by a uniaxial strain. Our findings provide an idea to create 2D magnetic semiconductors through metal-semiconductor transition in half metals.


Acute adaptations of the coronary circulation to exercise.
Coronary blood flow is tightly coupled to myocardial oxygen consumption to maintain a consistently high level of myocardial oxygen extraction. This tight coupling has been proposed to depend on periarteriolar oxygen tension, signals released from cardiomyocytes (adenosine acting on K+(ATP) channels), and/or the endothelium (prostanoids, nitric oxide, endothelin [ET]) and autonomic influences (catecholamines), but the contribution of each of these regulatory pathways and their interactions are still incompletely understood. Until recently, experimental studies into the regulation of coronary blood flow during exercise were principally performed in the dog. We have performed several studies on the regulation of vasomotor tone in coronary resistance vessels in chronically instrumented exercising swine. These studies have shown that the coronary resistance vessels in swine lack significant alpha-adrenergic control, but that these vessels are subject to beta-adrenergic feed-forward control during exercise, which is aided by a parasympathetic withdrawal. In addition, withdrawal of an ET-mediated vasoconstrictor influence also contributes to exercise-induced coronary vasodilation. Coronary blood flow regulation by endothelial and metabolic vasodilator pathways contributes to resting vasomotor tone regulation but does not appear to contribute to the exercise-induced coronary vasodilation. Furthermore, blockade of one vasodilator pathway is not compensated by an increased contribution of the other vasodilator mechanisms, suggesting that porcine coronary vasomotor control by endothelial and metabolic factors occurs in a linear additive rather than a nonlinear synergistic fashion.


Pervasive Observation Medicine: The Application of RFID to Improve Patient Safety in Observation Unit of Hospital Emergency Department.
Over the past decade, observation medicine has become an important component of emergency medicine. There are several settings in which observation medicine has been useful and valuable.(1) RFID as the patient identification, not only generates the on-line laboratory data and radiology report via hand-held wireless PDA, this RFID system help physician stream-line patient admission to acute bed or ICU in the emergency department more effectively.


MC1R expression in HaCaT keratinocytes inhibits UVA-induced ROS production via NADPH oxidase- and cAMP-dependent mechanisms.
Ultraviolet A (UVA) radiations are responsible for deleterious effects, mainly due to reactive oxygen species (ROS) production. Alpha-melanocyte stimulating hormone (α-MSH) binds to melanocortin-1 receptor (MC1R) in melanocytes to stimulate pigmentation and modulate cutaneous inflammatory responses. MC1R may be induced in keratinocytes after UV exposure. To investigate the effect of MC1R signaling on UVA-induced ROS (UVA-ROS) production, we generated HaCaT cells that stably express human MC1R (HaCaT-MC1R) or the Arg151Cys (R(151)C) non-functional variant (HaCaT-R(151)C). We then assessed ROS production immediately after UVA exposure and found that: (1) UVA-ROS production was strongly reduced in HaCaT-MC1R but not in HaCaT-R(151)C cells compared to parental HaCaT cells; (2) this inhibitory effect was further amplified by incubation of HaCaT-MC1R cells with α-MSH before UVA exposure; (3) protein kinase A (PKA)-dependent NoxA1 phosphorylation was increased in HaCaT-MC1R compared to HaCaT and HaCaT-R(151)C cells. Inhibition of PKA in HaCaT-MC1R cells resulted in a marked increase of ROS production after UVA irradiation; (4) the ability of HaCaT-MC1R cells to produce UVA-ROS was restored by inhibiting epidermal growth factor receptor (EGFR) or extracellular signal-regulated kinases (ERK) activity before UVA exposure. Our findings suggest that constitutive activity of MC1R in keratinocytes may reduce UVA-induced oxidative stress via EGFR and cAMP-dependent mechanisms.


Anomalous course of the recurrent motor branch of the median nerve in a patient with carpal tunnel syndrome.
A case is discussed in which carpal tunnel syndrome with an unusual course of the motor recurrent branch was discovered at operation. The importance of a careful dissection of the median nerve in this condition is emphasized.


Variables associated with perceived sleep disorders in methadone maintenance treatment (MMT) patients.
To characterize sleep disorders in methadone maintenance treatment (MMT) patients, we evaluated sleep quality of 101 non-selective patients from our MMT clinic in Israel between July, 2003 and July, 2004 by using the self-report questionnaire Pittsburgh Sleep Quality Index (PSQI). Patients' urine tests were analyzed for methadone metabolite, opiates, benzodiazepine, cocaine, cannabis and amphetamines. Their urine results for drug abuse throughout the months prior to filling in the questionnaire and their maintenance methadone doses were recorded. Drug abuse was defined by at least one positive urine test. Methadone serum levels were available in 55 patients, assessed by Gas Chromatography Mass Spectroscopy. The patients' self-reported chronic pain questionnaires and their diagnosed psychiatric disorders were analyzed. Out of the 101 study patients, 78.2% were male, 52.5% had psychiatric disorders, 46.5% reported having chronic pain and 46.5% had positive urine for benzodiazepine. The mean daily methadone dose was 157+/-52.9 mg. The mean PSQI score was 9+/-4.8 (75.2% had scores >5 indicating "poor sleepers"). PSQI scores were higher in patients with positive urine for benzodiazepine, chronic pain and psychiatric disorders and they correlated with years of opiate abuse before admission to MMT, and with the methadone dose (r=0.48, p<0.0005). The latter two also correlated with each other. The PSQI was not correlated with duration in MMT, gender, age, abuse of opiates, cannabis or cocaine. We concluded that sleep disorders should be evaluated and treated among MMT patients, particularly in those with psychiatric disorders, benzodiazepine abuse, chronic pain and high methadone dose.


Trichomonas vaginalis Weakens human Amniochorion in an in vitro model of premature membrane rupture.
Trichomonas vaginalis (TV) infection is associated with preterm rupture of membranes (PROM) and preterm birth. We evaluated the effects of TV growth and metabolism on preparations of human amniochorion to understand and characterize how TV may impair fetal-membrane integrity and predispose to PROM and preterm birth. Term fetal membranes were evaluated using an established in vitro fetal-membrane model. Fresh TV clinical isolates were obtained from pregnant women. The protozoa (5.0x10(5) to 1.5x10(6)/ml) were incubated with fetal membranes in modified Diamond's medium for 20 h at 37 degrees C in 5% CO2.The effects of fetal-membrane strength (bursting tension, work to rupture, and elasticity) were measured using a calibrated Wheatstone-bridge dynamometer. Tests were also performed to evaluate the effects of 1) inoculum size; 2) metronidazole (50 microg/ml); and 3) cell-free filtrate. The TV-induced membrane effects were 1) isolate variable; 2) inoculum dependent; 3) incompletely protected by metronidazole; and 4) mediated by both live organisms as well as protozoan-free culture filtrates. Six of 9 isolates significantly reduced the calculated work to rupture (P < or = 0.02); 7 of 9 reduced bursting tension; and 1 of 9 reduced elasticity. One isolate significantly increased the work to rupture and bursting tension (P < or = 0.002). In vitro incubation of fetal membranes with TV can significantly impair the measures of fetal-membrane strength. This model may be used to delineate the mechanisms of TV-induced membrane damage. This study suggests that there are enzyme-specific effects as well as pH effects.


Decreased ATP sensitivity of a K+ channel and enhanced vascular smooth muscle relaxation in genetically hypertensive rats.
To characterize an ATP-sensitive K+ channel at the single-channel and tissue level in the vascular smooth muscle of normotensive and genetically hypertensive rats. Age- and sex-matched Wistar-Kyoto (WKY) rats and stroke-prone spontaneously hypertensive rats (SHRSP) were used. Patch-clamp single-channel recording was used to measure K+ channel activity in dissociated tail artery cells. The effect of the K+ channel-opener diazoxide and the specific ATP-sensitive K+ channel antagonist glyburide on isometric force development in isolated tail artery strips was determined by a standard muscle bath technique. The concentration of ATP that caused half-maximal reduction in channel activity was greater in the SHRSP than in the WKY rats. Tail artery strips and cells from SHRSP were more sensitive to the effect of diazoxide on relaxation and channel activity, and less responsive to the effect of glyburide, than were those from WKY rats. The decreased ATP sensitivity of this K+ channel may partly compensate for the increased vascular reactivity in hypertension, and the change in this property of the channel may be responsible for the altered sensitivity to diazoxide and glyburide in SHRSP.


Long-Acting Growth Hormone Preparations - Current Status and Future Considerations.
Long-acting GH (LAGH) preparations are currently being developed in an attempt to improve adherence. The profile of GH action following administration of LAGH raises practical questions about clinical monitoring and long-term safety and efficacy of these new therapeutic agents. Recent literature and meeting proceedings regarding LAGH preparations are reviewed. Multiple LAGH preparations are currently at various stages of development, allowing for decreased GH injection frequency from daily to weekly, biweekly, or monthly. Following administration of LAGH, the serum peak and trough GH and IGF-I levels vary depending upon the mechanism used to prolong GH action. Randomized, controlled clinical trials of some LAGH preparations have reported non-inferiority compared with daily recombinant human GH (rhGH) for improved growth velocity and body composition in children and adults with GH deficiency (GHD), respectively. No significant LAGH-related adverse events have been reported during short-term therapy. Multiple LAGH preparations are proceeding through clinical development with some showing promising evidence of short-term clinical efficacy and safety in children and adults with GHD. The relationship of transient elevations of GH and IGF-I following administration of LAGH to efficacy and safety remain to be elucidated. For LAGH to replace daily rhGH in the treatment of individuals with GHD, a number of practical questions need to be addressed including methods of dose adjustment, timing of monitoring of IGF-I, safety, efficacy, and cost-effectiveness. Long-term surveillance of efficacy and safety of LAGH preparations will be needed to answer these clinically relevant questions.


The distribution of cadmium, copper, lead, zinc and calcium in the tissues of the earthworm Lumbricus rubellus sampled from one uncontaminated and four polluted soils.
The tissue distribution of Cu, Cd, Pb, Zn, and Ca in the earthworm Lumbricus rubellus living in non-polluted and heavy-metal polluted soils was investigated. Cd, Pb and Zn were primarily accumulated within the posterior alimentary canal. As the whole-worm Pb burden increased, the proportion of the metal accumulated within this tissue fraction increased. A similar pattern was found for Zn. By contrast, 70%-76% of the Cd burden was found in the posterior alimentary canal, irrespective of the whole-worm Cd content. The accumulation of Cd, Pb and Zn primarily in the posterior alimentary canal prevents dissemination of large concentrations of these metals into other earthworm tissues, and may thus represent a dextoxification strategy based on accumulative immobilisation. Cu was distributed fairly evenly in the tissue fractions investigated. There was no evidence of sequestration of this metal. The apparent lack of a detoxification strategy may contribute to the well-known susceptibility of earthworms to low environmental Cu concentrations. Indeed, earthworms from the site of highest soil Cu (Ecton) were markedly smaller than those from the other sites sampled. The highest Ca concentrations were found in the anterior alimentary canal, and were related to calciferous gland activity. A large proportion of Ca was also stored as a physiologically available pool in the posterior alimentary canal. Despite huge variations in soil Ca concentrations, the body wall Ca levels were fairly similar in L. rubellus from all the study sites. Thus, L. rubellus may become physiologically adapted to soils of exceptionally low Ca concentration. The observations are discussed in the context of the merits of analysing specific tissues, rather than whole organisms, for the purpose of monitoring metal bioaccumulation.


Comparative Analysis of Inpatient Opioid Consumption Between Different Surgical Approaches Following Single Level Lumbar Spinal Fusion Surgery.
Single-center retrospective cohort study. To evaluate inpatient MME administration associated with different lumbar spinal fusion surgeries. Patients ≥18 years of age with a diagnosis of Grade I or II spondylolisthesis, stenosis, degenerative disc disease or pars defect who underwent one-level Transforaminal Lumbar Interbody Fusion (TLIF) or one-level Anterior Lumbar Interbody Fusion (ALIF) or Lateral Lumbar Interbody Fusion (LLIF) through traditional MIS, anterior-posterior position or single position approaches between L2-S1. Outcome measures included patient demographics, surgical procedure and approach, perioperative clinical characteristics, incidence of ileus and inpatient MME. Statistical analysis included one-way ANOVA with a post-hoc Tukey Test and Kruskal-Wallis Test with post-hoc Mann-Whitney test. MME was calculated as per the Centers for Medicare and Medicaid Services and previous literature. Significance set at P < .05. Mean age differed significantly between MIS TLIF (55.6 ± 12.5 years) and all other groups (Open TLIF 57.1 ± 12.5, SP ALIF/LLIF 57.9 ± 9.9, TP ALIF/LLIF 50.9 ± 12.7, Open ALIF/LLIF 58.4 ± 15.5). MIS TLIF had the shortest LOS compared to all groups except SP ALIF/LLIF. Total MME was significantly different between MIS TLIF and Open ALIF/LLIF (172.5 MME vs 261.1 MME, P = .044) as well as MIS TLIF and TP ALIF/LLIF (172.5 MME vs 245.4 MME, P = .009). There were no significant differences in MME/hour and incidence of ileus between all groups. Patients undergoing MIS TLIF had lower inpatient opioid intake compared to TP and SP ALIF/LLIF, as well as shorter LOS compared to all groups except SP ALIF/LLIF. Thus, it appears that the advantages of minimally invasive surgery are seen in minimally invasive TLIFs.


The small trial problem.
Many randomized trials that aim to assess new or commonly used medical or surgical interventions have been so small that the validity of conclusions becomes questionable. We illustrate the small trial problem using the power calculation of five Cochrane-reviewed studies that compared vertebroplasty versus placebo interventions. We discuss some of the reasons why the statistical admonition not to dichotomize continuous variables may not apply to the calculation of the number of patients required for trials to be meaningful. Placebo-controlled vertebroplasty trials planned to recruit between 23 and 71 patients per group. Four of five studies used the standardized mean difference of a continuous pain variable (centimeters on the visual analog scale (VAS)) to plan implausibly small trials. What is needed is not a mean effect at the population level but a measure of efficacy at the patient level. Clinical practice concerns the care of individual patients that vary in many more respects than the variation around the mean of a single selected variable. The inference from trial to practice concerns the frequency of success of the experimental intervention performed one patient at a time. A comparison of the proportions of patients reaching a certain threshold is a more meaningful method that appropriately requires larger trials. Most placebo-controlled vertebroplasty trials used comparisons of means of a continuous variable and were consequently very small. Randomized trials should instead be large enough to account for the diversity of future patients and practices. They should offer an evaluation of a clinically meaningful number of interventions performed in various contexts. Implications of this principle are not specific to placebo-controlled surgical trials. Trials designed to inform practice require a per-patient comparison of outcomes and the size of the trial should be planned accordingly.


Cholescintigraphy in the diagnosis of the complications of laparoscopic cholecystectomy.
Cholescintigraphy has been used to diagnose postoperative bile leaks and obstructions after open cholecystectomy. This imaging technique has been found to be sensitive for detecting bile leaks that arise from cholecystectomy and defects in the filling and emptying of bile ducts. These observations can be applied to patients who are suspected of having bile duct injury after laparoscopic cholecystectomy. Cholescintigraphy can detect a variety of bile leaks including those arising from the gallbladder fossa, cystic duct, and the common bile duct. In addition, cholescintigraphy can be used for management of patients after stenting or bypass surgery, to assess bile flow and operative sites for any residual bile leaks. Postoperative bile duct obstructions can also be seen with cholescintigraphy. In this article we discuss a comprehensive postoperative strategy for the management and diagnosis of biliary defects after laparoscopic cholecystectomy with cholescintigraphy.


Methotrexate Polyglutamate Monitoring in Patients With Crohn's Disease.
Methotrexate is an efficacious immunosuppressant for induction and maintenance of remission in Crohn's disease. The goal of this pilot study was to determine whether total or individual methotrexate glutamate levels (MTXGlun ) in red blood cells correlate with disease activity and adverse events in Crohn's disease. A cross-sectional study was undertaken with 12 patients on a stable dose of 25 mg weekly methotrexate (oral or subcutaneous). Clinical disease activity was assessed by the Harvey-Bradshaw Index (HBI), and biologic disease activity was measured by inflammatory markers. Concentrations of individual MTXGlun levels were measured in red blood cells (RBCs) using high-performance liquid chromatography-mass spectrometry. No association was observed between RBC individual (MTXGlun ) or total methotrexate glutamate concentrations and clinical disease activity (HBI score) or inflammatory markers or adverse events. Although Crohn's disease patients in remission appeared to generally have higher RBC total longer-chain methotrexate polyglutamate (MTXGlu3+4+5 ) concentrations compared with those with active disease, a definitive association between RBC MTXGlu3+4+5 levels and clinical disease activity could not be established. Larger longitudinal studies in patients with diverse disease activity are needed to establish the value of MTXGlun levels as indicators of treatment efficacy and clinical outcome.


Epidemiology of travel-associated infections in Oman 1999-2013: A retrospective analysis.
The number of travelers in Oman has increased significantly in the last 2 decades with an increase in the expatriate population workforce leading to the emergence of infections related to travel. This paper aims to highlight the burden of travel-related infections in Oman. Our study is a descriptive record-based review and analysis of travel-associated diseases over a 14 year time period from 1999 to 2013. The data was sourced from the communicable disease surveillance system, and central public health laboratory results. From 1999 to 2013 there were a combined total of 7022 cases of cholera, chikungunya, dengue, filariasis, leptospirosis, meningococcal infection, poliomyelitis, measles, schistosomiasis, viral hepatitis (A), typhoid and para-typhoid reported to and subsequently investigated by the Department of Communicable Diseases. Among these cases, 558 (7.9%) were attributed to travel. Fifty percent of these patients were admitted to hospitals. Travel-associated infections account for about 8% of notifiable infections in Oman and have low mortality rate. However, some travel-associated infections are considered as a threat to polio eradication and measles elimination programs. Furthermore, some can cause outbreaks that can overwhelm the healthcare system.


[The teratogenicity of 13-cis-retinoic acid on the mandibular-otic anlagen of the mouse embryo: a histological and histochemical approach].
Oral administration of 400 mg/kg of 13-cis retinoic acid to 9 day pregnant mice gives rise to important maxillofacial malformations. The first manifestation of teratogenic effect is an increase of density of cell death arising in the dorsal part of the first two branchial arches at day 9.5. These two arches become hypoplastic at days 10 and 11, and the preskeletal anlagen appear too late in comparison to control embryos. Meckel's cartilage is too curvilinear and medially situated. Pre-ossicular and pre-mandibular blastemata develop with spatial distortions which are well analyzable at days 16 and 17. They give some arguments to discuss several features of normal early development of this area.


Adhesion-blocking antibodies prepared against gp150 react with gp80 of Dictyostelium.
A membrane glycoprotein of 150 000 D, gp150, has been implicated in the mechanism of cell-cell adhesion which arises during development of Dictyostelium discoideum. This conclusion was founded on the observation that monovalent Fab' fragments prepared from an antiserum raised against partially purified gp150 are able to block cell-cell adhesion. We show that this serum contains antibodies to a distinct membrane glycoprotein, gp80, previously implicated in cell-cell adhesion. Reaction of Fab' to this surface molecule can account for the adhesion-blocking activity in the antiserum to gp 150. Moreover, binding of gp80 neutralized Fab' to gp150 does not block adhesion. If gp150 carries other determinants which bind adhesion-blocking Fab', these determinants must also be present on gp80. Thus, it is not clear that gp150 is directly involved in cell-cell adhesion of Dictyostelium.


Serum and saliva total tau protein as a marker for relapsing-remitting multiple sclerosis.
Multiple sclerosis (MS) is considered as motor and sensory function loss, which results from demyelination and following axonal lesion. Tau protein interferes in the construction and stabilization of microtubules that are needed for axonal transport. We hypothesize that the level of tau protein alters in MS, so we examined the level of total tau protein in serum, stimulated and unstimulated saliva as a suitable biomarker for detecting of relapsing-remitting MS. A case-control study was carried out in 30 healthy women and 30 who suffer MS and were hospitalized in Imam Reza hospital, Tehran, Iran. Total tau protein level was assayed in serum and stimulated and unstimulated whole saliva. Total tau protein was expressed at a lower level in serum of MS patients compared to control groups. The serum total tau protein level negatively correlated with EDSS. There was no significant difference in saliva total tau protein between MS and healthy individuals. Moreover, there was no significant correlation between saliva and serum total tau protein and between saliva total tau protein and EDSS. The serum level of tau protein is lower in MS and it may be considered as a potential biomarker in Multiple sclerosis. However, it seems that tau protein in the saliva isn't a suitable biomarker for detection of MS.


Seronegative bacteremic melioidosis caused by Burkholderia pseudomallei with ambiguous biochemical profile: clinical importance of accurate identification by 16S rRNA gene and groEL gene sequencing.
An aerobic gram-negative bacterium was isolated from the blood and sputum of an 84-year-old, chair-bound nursing home resident with acute bacteremic pneumonia. Although the phenotypic characteristics suggested that the bacterium could be Burkholderia pseudomallei, the Vitek 1 system (GNI+), which can successfully identify 99% of B. pseudomallei strains, showed that the bacterium was "unidentified." Immunoglobulin G against the lipopolysaccharide (LPS) of B. pseudomallei, as detected by an LPS-based enzyme-linked immunosorbent assay with 95% sensitivity, was negative in both the acute-phase and convalescent-phase sera. Sequencing of the groEL gene showed that the isolate was B. pseudomallei. Proper identification of the bacterium in this study is crucial, since there would be a radical difference in the duration of antimicrobial therapy.


Comparison of the decompressive effect of different surgical procedures for dysthyroid optic neuropathy using 3D printed models.
To compare the decompressive effect around the optic nerve canal among 3 different decompression procedures (medial, balanced, and inferomedial) using 3D printed models. In this experimental study, based on data obtained from 9 patients (18 sides) with dysthyroid optic neuropathy, a preoperative control model and 3 plaster decompression models were created using a 3D printer (total, 72 sides of 36 models). A pressure sensor was placed at the optic foramen, and the orbital space was filled with silicone. The surface of the silicone was pushed down directly, and changes in pressure were recorded at 2-mm increments of pushing. At 10 mm of pushing, there was significantly lower pressure in the medial (19,782.2 ± 4319.9 Pa, P = 0.001), balanced (19,448.3 ± 3767.4 Pa, P = 0.003), and inferomedial (15,855.8 ± 4000.7 Pa, P < 0.001) decompression models than in the control model (25,217.8 ± 6087.5 Pa). Overall, the statistical results for each 2-mm push were similar among the models up to 10 mm of pushing (P < 0.050). At each push, inferomedial decompression caused the greatest reduction in pressure (P < 0.050), whereas there was no significant difference in pressure between the medial and balanced decompression models (P > 0.050). All 3 commonly performed decompression procedures significantly reduced retrobulbar pressure. Because inferomedial decompression models obtained the greatest reduction in pressure on the optic nerve canal, inferomedial decompression should be considered the most reliable procedure for rescuing vision in dysthyroid optic neuropathy.


Denoising based on spatial filtering.
We present a method for removing unwanted components of biological origin from neurophysiological recordings such as magnetoencephalography (MEG), electroencephalography (EEG), or multichannel electrophysiological or optical recordings. A spatial filter is designed to partition recorded activity into stimulus-related and stimulus-unrelated components, based on a criterion of stimulus-evoked reproducibility. Components that are not reproducible are projected out to obtain clean data. In experiments that measure stimulus-evoked activity, typically about 80% of noise power is removed with minimal distortion of the evoked response. Signal-to-noise ratios of better than 0 dB (50% reproducible power) may be obtained for the single most reproducible spatial component. The spatial filters are synthesized using a blind source separation method known as denoising source separation (DSS) that allows the measure of interest (here proportion of evoked power) to guide the source separation. That method is of greater general use, allowing data denoising beyond the classical stimulus-evoked response paradigm.


ZBTB2 links p53 deficiency to HIF-1-mediated hypoxia signaling to promote cancer aggressiveness.
Aberrant activation of the hypoxia-inducible transcription factor HIF-1 and dysfunction of the tumor suppressor p53 have been reported to induce malignant phenotypes and therapy resistance of cancers. However, their mechanistic and functional relationship remains largely unknown. Here, we reveal a mechanism by which p53 deficiency triggers the activation of HIF-1-dependent hypoxia signaling and identify zinc finger and BTB domain-containing protein 2 (ZBTB2) as an important mediator. ZBTB2 forms homodimers via its N-terminus region and increases the transactivation activity of HIF-1 only when functional p53 is absent. The ZBTB2 homodimer facilitates invasion, distant metastasis, and growth of p53-deficient, but not p53-proficient, cancers. The intratumoral expression levels of ZBTB2 are associated with poor prognosis in lung cancer patients. ZBTB2 N-terminus-mimetic polypeptides competitively inhibit ZBTB2 homodimerization and significantly suppress the ZBTB2-HIF-1 axis, leading to antitumor effects. Our data reveal an important link between aberrant activation of hypoxia signaling and loss of a tumor suppressor and provide a rationale for targeting a key mediator, ZBTB2, to suppress cancer aggressiveness.


Opposite regulation of cAMP concentration in the quail oviduct and the mouse uterus by tamoxifen. Correlation with estrogen-antagonist and estrogen-agonist activity.
The ability of estradiol and tamoxifen to regulate cAMP levels and cAMP phosphodiesterase activities has been determined in the quail oviduct and in the mouse uterus. In the quail, tamoxifen (1 mg/kg daily for 3 days) had no effect on oviducal growth but significantly increased cAMP concentration (+49%). Injected concurrently with estradiol, tamoxifen completely inhibited oviduct growth as well as the increase of cAMP phosphodiesterase activity induced by the hormone alone and increased cAMP concentration (+229% over estradiol treated group). In the mouse, estradiol and tamoxifen displayed uterotrophic activity and increased cAMP phosphodiesterase activity. In both groups, cAMP concentration was greatly reduced (-76% in estradiol treated group; -86% in tamoxifen treated group). The opposite regulation of cAMP levels in the quail oviduct and the mouse uterus by tamoxifen reflected large differences in the contribution of calmodulin-dependent and -independent forms of phosphodiesterase to the hydrolysis of cAMP in the two models and the fact that tamoxifen stimulated the activity of the calmodulin-independent isoenzyme, while it competitively inhibited the activation of the calmodulin-dependent isoenzyme by calmodulin. Several lines of evidence strongly suggest that the regulation of cAMP levels is involved in growth-inhibiting or growth-promoting activity of tamoxifen.


Advancing Breast Cancer Heterogeneity Analysis: Insights from Genomics, Transcriptomics and Proteomics at Bulk and Single-Cell Levels.
Breast cancer continues to pose a significant healthcare challenge worldwide for its inherent molecular heterogeneity. This review offers an in-depth assessment of the molecular profiling undertaken to understand this heterogeneity, focusing on multi-omics strategies applied both in traditional bulk and single-cell levels. Genomic investigations have profoundly informed our comprehension of breast cancer, enabling its categorization into six intrinsic molecular subtypes. Beyond genomics, transcriptomics has rendered deeper insights into the gene expression landscape of breast cancer cells. It has also facilitated the formulation of more precise predictive and prognostic models, thereby enriching the field of personalized medicine in breast cancer. The comparison between traditional and single-cell transcriptomics has identified unique gene expression patterns and facilitated the understanding of cell-to-cell variability. Proteomics provides further insights into breast cancer subtypes by illuminating intricate protein expression patterns and their post-translational modifications. The adoption of single-cell proteomics has been instrumental in this regard, revealing the complex dynamics of protein regulation and interaction. Despite these advancements, this review underscores the need for a holistic integration of multiple 'omics' strategies to fully decipher breast cancer heterogeneity. Such integration not only ensures a comprehensive understanding of breast cancer's molecular complexities, but also promotes the development of personalized treatment strategies.


Abacillary pulmonary tuberculosis.
In Copenhagen, a city with a low incidence of tuberculosis, 72 patients with discrete pulmonary infiltrates on the chest X-ray had a tentative diagnosis of tuberculosis. All were sputum smear negative for acid-fast bacilli. A prospective randomised study was carried out to determine whether these patients would benefit from chemotherapy. Out of the 72 patients, 22 (30.6%) had positive cultures initially and were treated. Of the remaining 50 patients, 22 received chemotherapy and 28 were untreated controls. Among the untreated patients 8 (29%) later became culture positive for M. tuberculosis, whereas none of the treated patients developed culture positive tuberculosis during the follow-up period of 5 years.


Demographic changes and the development of competition in a native Australian eucalypt forest invaded by exotic pines.
Data from the second half of a 14-year study of a eucalypt dry sclerophyll forest invaded by Pinus radiata D. Don shows further progress in the development of a mixed eucalypt-pine stand. Earlier (Chilvers and Burdon 1983) it was clear that pines invading from an adjacent commercial plantation were starting to contribute their own progeny to the mixed stand. Initially the recruitment of established pines (> 1 m high) from these seedlings was very slow. However, since 1984, increasing numbers of seedlings have survived beyond their first year, suggesting the possibility of another wave of successful establishment in the near future. The established fraction of the pine population continued to grow rapidly and incurred no mortality. Many of these plants have now emerged through the top of the eucalypt canopy. In contrast, both juvenile and adult eucalypt numbers continued to decline. Despite these demographic and visual changes, comparative analysis of the growth of established pines over two consecutive 7-year periods (1974-1981; 1981-1988) showed that competition between neighbouring plants, especially pines, is beginning to affect individual growth rates. Growth rates of pines having other pines as nearest neighbours declined 40% between 1974-1981 and 1981-1988, while that of pines with eucalypts as nearest neighbours fell by 28%. In pine-pine nearest neighbour (NN) pairs, growth rates were significantly affected by the size (diameter) of the nearest neighbour. In pine-eucalypt NN pairs growth rates of the pines were not affected by the size of the eucalypt, but these were significantly negatively correlated with the inverse of the distance to the eucalypt. More broad-based assessment of 43 pines showed a significant relationship between their growth rates and the summed competitive effect of other pines within a 5-m radius. No such relationship was found between growth rate and the summed competitive effect of eucalypts in the 5-m zone.


Corrigendum: Plastid Genomes of Five Species of Riverweeds (Podostemaceae): Structural Organization and Comparative Analysis in Malpighiales.
[This corrects the article DOI: 10.3389/fpls.2019.01035.].


Unruptured brain arteriovenous malformations causing seizures localize to one common brain network.
Seizures are a frequent symptom of unruptured brain arteriovenous malformations (bAVMs). However, the brain regions responsible for these seizures remain unclear. To identify the brain regions causally involved in bAVM-related seizures, we retrospectively reviewed 220 patients with unruptured bAVMs. Using voxel-based lesion-symptom mapping (VLSM) analyses, we tested whether individual brain regions were associated with unruptured bAVM-related seizures. The result revealed that unruptured bAVMs causing seizures are anatomically heterogeneous at the voxel level. Subsequently, lesion network mapping (LNM) analyses was performed to determine whether bAVMs causing seizures belonged to a distributed brain network. LNM analyses indicated that these lesions were located in a functional network characterized by connectivity to the left caudate and precuneus. Moreover, the discrimination performance of the identified seizure network was evaluated in discovery set by calculating the individualized network damage score and was tested in validation set. Based on the calculated network damage scores, patients were divided into low-, medium-, and high-risk groups. The prevalence of seizures significantly differed among the three risk categories in both discovery (p = .003) and validation set (p = .004). Finally, we calculated the percentage of voxels in the canonical resting-state networks that overlapped with the seizure-susceptible brain regions to investigate the involvement of resting-state networks. With an involvement percentage over 50%, the frontoparietal control (82.9%), limbic function (76.7%), and default mode network (69.3%) were considered to be impacted in bAVM-related seizures. Our study identified the seizure-susceptible brain regions for unruptured bAVMs, which could be a plausible neuroimaging biomarker in predicting possible seizures.


Evaluation on lifestyle and nutrition among Hungarian elderly.
A cross-sectional study on nutritional habits and lifestyle on 250 elderly Hungarian people was performed. The education level, marital status, housing situation, the time for social and outdoor activities, smoking and drinking habits and the general health condition were evaluated.Some anthropometric parameters, the nutritional habits, daily meal patterns and food preference were also briefly analyzed. The overweight, lack of physical activity and unfavourable nutritional habits were the main observations on this population. These trends should be turned by stronger medical intervention based on primary care.


Properties of the viruses selected during persistent infection of L cells with VSV.
Two virus clones, VSV-mp and VSV-sp, were isolated from L cells persistently infected with VSV (New Jersey serotype). Both clones were more temperature sensitive than the parent virus, VSV-o, and grew more slowly, gave smaller plaques, less c.p.e. and lower virus yields in L cells. Unlike the parent virus, both persistent viruses induced interferon production in L cells. Stable carrier cultures could be obtained from L cells infected with VSV-sp at low multiplicities without pretreatment with interferon. This may be related to the fact that VSV-sp is more sensitive to interferon than either VSV-mp or VSV-o.


[Effects of fungus on the growth of Dendrobium candidum and D. nobile in vitro culture].
To examine effects of fungus AR-18 (Epulorhiza sp.) on the growth of Dendrobium candidum and D. nobile in vitro culture. Effects of fungus AR-18 on fresh weight and dry weight of D. candidum and D. nobile were studied in vitro culture, when agar was used as rest, and effects of fungus AR-18 and nutrients on fresh weight and dry weight of D. candidum and D. nobile were studied in vitro culture, when vermiculite was used as rest. In agar medium, effects of fungus AR-18 on fresh weight and dry weight of D. candidum and D. nobile were not significant (P > 0.05). However, in vermiculite medium, the effect of fungus AR-18 on fresh weight of D. nobile was significant (P < 0.05). Fresh weight of D. nobile inoculated with fungus AR-18 was increased by 16% compared with the non-inoculated ones (control). The effect of fungus AR-18 on dry weight of D. nobile was very significant (P < 0.01). Dry weight of D. nobile inoculated with fungus AR-18 was increased by 21% compared with the control. In vermiculite medium, the effect of combination treatment (Fungus AR-18 + Distilled H2O) on fresh weight of D. nobile was very significant (P < 0.01). Fresh weight of D. nobile inoculated with fungus AR-18 was increased by 47% compared with the control, while the effect of combination treatment (Fungus AR-18 + Nutrients) on fresh weight of D. nobile was not significant (P > 0.05). In addition, the effect of combinations treatment (Fungus AR-18 + Nutrients) and (Fungus AR-18 + Distilled H2O) on dry weight of D. nobile was not significant (P > 0.05). So the combination treatment (Fungus AR-18 + Distilled H2O) was the best for the growth of D. nobile. In vermiculite medium, effects of fungus AR-18 and nutrients on fresh weight and dry weight of D. candidum were not significant (P > 0.05). It is important in vitro culture to select a proper rest for the control of the growth of fungus to establish a beneficial symbiotic relationship between plants and fungi. If the fungus AR-18 is utilized as biological manure in got-up planting of D. nobile, a good result may be obtained.


Mammalian artificial chromosomes: A review.
A mammalian artificial chromosome (MAC) may be assembled through the juxtapposition of three kinds of DNA elements: a centromere, several DNA replication origins, and two telomeric repeats. The resulting structure should be able to carry and express one or more selected genes (transgenes), introduced for specific purposes. The minimal length is unknown, but may be of several Mb.Of its basic elements, the telomeres may present lesser problems, in view of their simple composition and organization. Centromeres could be an issue, given their many unknowns. Mammalian DNA replication origins are at present poorly characterized, but it is expected that at least one may be contained within the MAC components, especially the transgene. Their overall assembly may require a combination of in vivo and in vitro approaches.A promising strategy aims at constructing two telomeric arms of a MAC, one of which may include the transgene. The two novel arms could acquire a functional centromere through recombination with the two arms of a resident chromosome. Alternatively, if the two telomeric constructs are also endowed with properly placed and oriented centromeric sequences, a centromere may be rescued in vivo by homologous recombination with the external parts of the centromere of the resident chromosome. Positive selection for the artificial arms and counterselection against the resident arms should facilitate the assembly process.The assembly of such construct would not change the ploidy number of the host cell. After loading of a transgene, however, the resulting MAC may be isolated and transferred into an expression cell, where it may represent a novel chromosomal element. In this case untoward effects to the host cell may derive from an ensuing dosage effect for the transgene(s) rather than from the presence of a MAC per se.A MAC may contribute to a deeper understanding of the structural requirements for chromosomal function and evolution as well as the mechanism of chromatin formation. It should also help in the development of second generation vectors for transfer of Mb-long DNA sequences, as required for properly regulated mammalian gene function as well as, possibly, for therapy.


Regulatory effect of 1,25-dihydroxycholecalciferol on calcium fluxes in thyroid FRTL-5 cells.
The aim of the present study was to investigate the effect of 1,25-dihydroxycholecalciferol (1,25(OH)2-D3) on the regulation of calcium fluxes in rat thyroid FRTL-5 cells. The ATP-induced uptake of 45Ca2+ was decreased in cells pretreated with 1,25(OH)2D3 for 48 h. No effect was seen on basal uptake of 45Ca2+. At least a 24 h incubation period was required for the effect of 1,25(OH)2D3 to be expressed. Pretreatment with 1,25(OH)2D3 for 48 h did not change resting intracellular Ca2+ ([Ca2+]i) in fura-2-loaded FRTL-5 cells. However, the ATP-induced increase in [Ca2+]i was significantly enhanced in cells preincubated with 1,25(OH)2D3. The effect of 1,25(OH)2D3 was abolished in Ca(2+)-free buffer. No difference in the ionomycin-induced increase in [Ca2+]i was observed between control cells and cells pretreated with 1,25(OH)2D3. However, in Ca(2+)-free buffer the ionomycin response was decreased in cells incubated with 1,25(OH)2D3. The ATP-induced change in [Ca2+]i was decreased when ATP was added after ionomycin to cells treated with 1,25(OH)2D3. The results suggest that 1,25(OH)2D3 has a regulatory effect on Ca2+ fluxes in FRTL-5 cells, possibly by acting on Ca2+ sequestration.


DNA oxidative damage in mammalian spermatozoa: where and why is the male nucleus affected?
Gamete DNA integrity is one key parameter conditioning reproductive success as well as the quality of life for the offspring. In particular, damage to the male nucleus can have profound negative effects on the outcome of fertilization. Because of the absence of repair activity of the quiescent mature spermatozoa it is easily subjected to nuclear damage, of which oxidative damage is by far the most prominent. In relation to the organization of the mammalian sperm nucleus we show here that one can correlate the nuclear regions of lower compaction with areas preferentially showing oxidative damage. More precisely, we show that oxidative DNA damage targets primarily histone-rich and nuclear matrix-attached domains located in the peripheral and basal regions of the mouse sperm nucleus. These particular sperm DNA domains were recently shown to be enriched in genes of paramount importance in postfertilization DNA replication events and in the onset of the embryonic developmental program. We propose that monitoring of sperm DNA oxidation using the type of assay presented here should be considered in clinical practice when one wants to estimate the integrity of the paternal nucleus along with more classical assays that essentially analyze DNA fragmentation and nucleus compaction.


Beyond phylotyping: understanding the impact of gut microbiota on host biology.
Microbial constituents of the gut microbiome interact with each other and the host to alter the luminal environment and impact development, motility, and homeostasis of the gut. Powerful methods are becoming available to investigate connections between the gut microbiome and human health. While high-throughput sequencing of 16S rRNA genes can be used to identify and enumerate microbes in the gut, advances in several techniques (e.g., metagenomics, metatranscriptomics, metabolomics, and metaproteomics) are providing a clearer view as to the specific activities of the microbiota in the context of functional host-microbe interactions. Testing emergent hypotheses regarding microbial effects on host biology, which arise from analyses of 'Big Data' generated from massive parallel high-throughput sequencing technology and spectroscopic techniques, to guide translational research is an important goal for the future. Insights regarding the fundamental operating principles of the gut microbiota should lay the foundation for rational manipulation of the microbiota to promote human health. In this review, we provide an overview of current research on the gut microbiome emphasizing current state-of-the-art technologies, approaches, and directions for improvement of our understanding of the impact of the gut microbiota with specific focus on gastrointestinal motility disorders.


Carnitine synthesis and uptake into cells are stimulated by fasting in pigs as a model of nonproliferating species.
In rodents, fasting increases the carnitine concentration in the liver by an up-regulation of enzymes of hepatic carnitine synthesis and novel organic cation transporter (OCTN) 2, mediated by activation of peroxisome proliferator-activated receptor (PPAR) alpha. This study was performed to investigate whether such effects occur also in pigs which like humans, as nonproliferating species, have a lower expression of PPARalpha and are less responsive to treatment with PPARalpha agonists than rodents. An experiment with 20 pigs was performed, which were either fed a diet ad-libitum or fasted for 24 h. Fasted pigs had higher relative mRNA concentrations of the PPARalpha target genes carnitine palmitoyltransferase 1 and acyl-CoA oxidase in liver, heart, kidney, and small intestinal mucosa than control pigs, indicative of PPARalpha activation in these tissues (P<.05). Fasted pigs had a higher activity of gamma-butyrobetaine dioxygenase (BBD), enzyme that catalyses the last step of carnitine biosynthesis in liver and kidney, and higher relative mRNA concentrations of OCTN2, the most important carnitine transporter, in liver, kidney, skeletal muscle, and small intestinal mucosa than control pigs (P<.05). Fasted pigs moreover had higher concentrations of free and total carnitine in liver and kidney than control pigs (P<.05). This study shows for the first time that fasting increases the activity of BBD in liver and kidney and up-regulates the expression of OCTN2 in various tissues of pigs, probably mediated by PPARalpha activation. It is concluded that nonproliferating species are also able to cover their increased demand for carnitine during fasting by an increased carnitine synthesis and uptake into cells.


Altered liver gene expression in CCl4-cirrhotic rats is partially normalized by insulin-like growth factor-I.
We have previously shown that the administration of low doses of insulin-like growth factor-I (IGF-I) to CCl4-cirrhotic rats improves liver function and reduces fibrosis. To better understand the mechanisms behind the hepatoprotective effects of IGF-I, and to identify those genes whose expression is affected in cirrhosis and after IGF-1 treatment, we have performed differential display of mRNA analysis by means of polymerase chain reaction (PCR) in livers from control and CCl4-cirrhotic rats treated or not with IGF-I. We have identified 16 genes that were up- or down-regulated in the cirrhotic liver. IGF-I treatment partially normalized the expression of eight of these genes, including serine proteinase inhibitors such as serpin-2 and alpha-1-antichymotripsin, alpha-1-acid glycoprotein, and alpha-2u-globulin. Additionally, we show that IGF-I enhanced the regenerative activity in the cirrhotic liver, as determined by the increased expression of the proliferating cell nuclear antigen (PCNA). Finally, IGF-I treatment partially restored the expression of growth hormone receptor (GHR) and the levels of global genomic DNA methylation, which are reduced in human and experimental cirrhosis. Taken together, our observations confirm the hepatoprotective effects of IGF-I, and suggest that this action can be exerted in part through the normalization of liver gene expression, growth hormone (GH) responsiveness and global genomic DNA methylation.


Addressing COVID-19 inequities using bidirectional crisis and emergency risk communication and vaccine clinic interventions: a descriptive study.
Im/migrants (immigrants and migrants, including refugees, asylum seekers, and individuals without legal documentation) experience unique assets and needs in relation to coronavirus disease 2019 (COVID-19). Community-based participatory research (CBPR) is one way to engage im/migrant communities. Rochester Healthy Community Partnership (RHCP) is a CBPR partnership in Rochester, Minnesota. RHCP partners noted that credible COVID-19 information was not available to their communities. In response, RHCP formed a COVID-19 Task Force and adapted the Centers for Disease Control and Prevention's Crisis and Emergency Risk Communication (CERC) framework to create an intervention that prioritized im/migrant groups experiencing health disparities. In the CERC intervention, communication leaders delivered COVID-19 health messages to their social networks and documented related concerns. RHCP relayed these concerns to regional leaders to ensure that im/migrant experiences were included in decision making. Once vaccines were available, RHCP continued to deploy the CERC intervention to promote vaccination equity. The aims of this paper are to (1) describe the implementation of a bidirectional CERC intervention for vaccination equity, and (2) describe a community-engaged and community-based vaccine clinic intervention. First, we surveyed participants (n = 37) to assess COVID-19 experiences, acceptability of the CERC intervention, and motivation to receive a COVID-19 vaccination. Second, we collaborated with community partners to hold vaccine clinics. We report descriptive statistics from each intervention. When asked about the acceptability of the CERC intervention for vaccine equity, most participants either reported that they 'really liked it' or 'thought it was just ok'. Most participants stated that they would recommend the program to family or friends who have not yet received the COVID-19 vaccine. Almost all participants reported that they felt 'much more' or 'somewhat more' motivated to receive a COVID-19 vaccine after the intervention. We administered 1158 vaccines at the vaccination clinics. We found that participants viewed the CERC intervention for vaccination equity as an acceptable way to disseminate COVID-19-related information. Nearly all participants reported that the intervention convinced them to receive a COVID-19 vaccine. In our experience, community-engaged and community-based clinics are a successful way to administer vaccines to im/migrant communities during a pandemic.


Multi-resolution geometric modeling of the mitral heart valve leaflets.
An essential element of cardiac function, the mitral valve (MV) ensures proper directional blood flow between the left heart chambers. Over the past two decades, computational simulations have made marked advancements toward providing powerful predictive tools to better understand valvular function and improve treatments for MV disease. However, challenges remain in the development of robust means for the quantification and representation of MV leaflet geometry. In this study, we present a novel modeling pipeline to quantitatively characterize and represent MV leaflet surface geometry. Our methodology utilized a two-part additive decomposition of the MV geometric features to decouple the macro-level general leaflet shape descriptors from the leaflet fine-scale features. First, the general shapes of five ovine MV leaflets were modeled using superquadric surfaces. Second, the finer-scale geometric details were captured, quantified, and reconstructed via a 2D Fourier analysis with an additional sparsity constraint. This spectral approach allowed us to easily control the level of geometric details in the reconstructed geometry. The results revealed that our methodology provided a robust and accurate approach to develop MV-specific models with an adjustable level of spatial resolution and geometric detail. Such fully customizable models provide the necessary means to perform computational simulations of the MV at a range of geometric accuracies in order to identify the level of complexity required to achieve predictive MV simulations.


The beta chorionic gonadotropin-beta luteinizing gene cluster maps to human chromosome 19.
We used a cloned human cDNA probe homologous to the placenta chorionic gonadotropin beta subunit (CGB) and to the pituitary luteinizing hormone beta subunit (LHB) and Southern blotting techniques to analyse DNA from a series of rodent X human somatic cell hybrids for the presence of specific gonadotropin beta subunit related sequences. Our results provide evidence for the assignment and linkage of the eight genes (or pseudogenes) coding for the beta subunit of these glycoprotein hormones to chromosome 19. Moreover, we observed a strict concordance between the permissivity of mouse X man hybrid cells to enteroviruses (which is linked to the presence of specific cell receptors encoded by human chromosome 19) and the presence of CGB and LHB related sequences, thus confirming the localization of the structural genes for the beta subunits on chromosome 19.


Intratumoral heterogeneity identified at the epigenetic, genetic and transcriptional level in glioblastoma.
Heterogeneity is a hallmark of glioblastoma with intratumoral heterogeneity contributing to variability in responses and resistance to standard treatments. Promoter methylation status of the DNA repair enzyme O(6)-methylguanine DNA methyltransferase (MGMT) is the most important clinical biomarker in glioblastoma, predicting for therapeutic response. However, it does not always correlate with response. This may be due to intratumoral heterogeneity, with a single biopsy unlikely to represent the entire lesion. Aberrations in other DNA repair mechanisms may also contribute. This study investigated intratumoral heterogeneity in multiple glioblastoma tumors with a particular focus on the DNA repair pathways. Transcriptional intratumoral heterogeneity was identified in 40% of cases with variability in MGMT methylation status found in 14% of cases. As well as identifying intratumoral heterogeneity at the transcriptional and epigenetic levels, targeted next generation sequencing identified between 1 and 37 unique sequence variants per specimen. In-silico tools were then able to identify deleterious variants in both the base excision repair and the mismatch repair pathways that may contribute to therapeutic response. As these pathways have roles in temozolomide response, these findings may confound patient management and highlight the importance of assessing multiple tumor biopsies.


Human amylin induces CD4+Foxp3+ regulatory T cells in the protection from autoimmune diabetes.
Autoimmune diabetes is a disorder of immune homeostasis that leads to targeted insulin-secreting islet β cell destruction characterized by insulitis. Human amylin (hA) is an important neuroendocrine hormone co-secreted with insulin by pancreatic β cells. Here, we report hA immune-modulatory action through inducing regulatory T cells. We ex vivo-treated human peripheral blood mononuclear cells (hPBMCs) with hA for 24 h and counted CD4+Foxp3+ regulatory T cells (Treg) using flow cytometry. Diabetic status was monitored and splenic Treg were measured in non-obese diabetic (NOD) male mice. NOD mice were intraperitoneally injected once daily with hA (n = 25) or solvent for control (n = 25) for 7 months continuously. Spleen tissues were collected at the end of intervention and processed for flow cytometry and Western blot. We found a 2.9-fold (p < 0.05) increase of CD4+Foxp3+ Treg in hPBMCs treated with 10 nmol/L hA compared with negative control. Incidence of diabetes in hA-treated NOD mice decreased 44% (p = 0.045) in the 6th month and 57% (p = 0.0002) in the 7th month. Meanwhile, the hA treatment induced a 1.5-fold increase of CD4+Foxp3+ Treg from mouse splenocytes (p = 0.0013). Expression of transforming growth factor-β (TGF-β) and toll-like receptor-4 (TLR-4) were upregulated in hA-treated mice. Human amylin might protect against autoimmune diabetes via the induction of CD4+Foxp3+ Treg, which suggests a novel approach to improve autoimmune conditions.


Proposal of a new method for the risk scoring and categorization of Brazilian food services.
Risk scores are used worldwide to predict foodborne disease (FBD) outbreaks in the food service industry. This study aims to develop and validate a new method for the calculation of the FBD risk score for the checklist used to categorize food service outlets. The proposed novel method is based on a risk score for each item using a risk matrix (consequence × probability), overcoming the limitations of the previous scoring process used during the World Cup in Brazil, which was based on a factorial analysis. The classification of consequences was based on critical points identified by experts prior to the World Cup in Brazil. Probability was defined based on the violation percentage of each item evaluated during inspections from 1536 food service outlets. Validation was performed using a secondary database of 3072 food service assessments in two inspection cycles. The risk scores of the new method were compared with those used during the World Cup. Each food service was classified based on their risk score into four categories: A, B, C, and pending. Good concordance (Lin's correlation coefficient = 0.8711 and 0.9205) was observed between the new and previous scores in the two inspection cycles, respectively. Comparison of the classifications showed substantial agreement (Kappa = 0.749, p < 0.001) to the first inspection cycle and near-perfect agreement (Kappa = 0.821; p < 0.001) to the second inspection cycle. This new method allows the inclusion and exclusion of assessment elements depending on local reality. Simpler methods can be used throughout Brazil and serve as a model for other countries' food safety assessments.


Global chimeric exchanges within the intracellular face of the bradykinin B2 receptor with corresponding angiotensin II type Ia receptor regions: generation of fully functional hybrids showing characteristic signaling of the AT1a receptor.
The intracellular (IC) face of the G-protein coupled receptors (GPCR), bradykinin (BK) B2 and angiotensin (AT) 1a, is similar in sequence homology and in size. Both receptors are known to link to Galphai and Galphaq but differ markedly in a number of physiologic actions, particularly with respect to their hemodynamic action. We made single as well as multiple, global replacements within the IC of BKB2R with the corresponding regions of the AT1aR. When stably transfected into Rat-1 cells, these hybrid receptors all bound BK with high affinity. Single replacement of the intracellular loop 2 (IC2) or the distal 34 residues of the C-terminus (dCt) with the corresponding regions of AT1aR resulted in chimera, which turned over phosphotidylinositol (PI) and released arachidonic acid (ARA) as WT BKB2R. In contrast, incorporation of the AT1aR IC3 in a single replacement abolished signal transduction. However, the simultaneous exchange of IC2 and IC3 of BKB2R with AT1aR resulted in a receptor responding to BK with PI turnover and ARA release approximately 4-fold greater than WT BKB2R. Likewise, the simultaneous replacement of IC2 and dCt resulted in a 2.8- and 1.6-fold increase in PI turnover and ARA release, respectively. In contrast, the dual replacement of IC3 and dCt could not overcome the deleterious effects of the IC3 replacement, resulting in very low PI activation and ARA release. Replacement of all three IC domains (IC2, IC3, and dCt) resulted in PI closer to that of AT1aR than BKB2R. The uptake of the receptor chimeras was similar to that of WT BKB2R with the exception of the IC3/dCt dual mutant, which exhibited very poor internalization (18% at 60'). When transfected into Rat-1 cells, the AT1aR markedly increased the expression of connective tissue growth factor (CTGF) mRNA, while BK slightly decreased it. The dual IC2/dCt and triple IC2/IC3/dCt hybrids both upregulated CTGF mRNA in response to BK. These results show that the IC face of the BKB2R can be exchanged with that of AT1aR, producing hybrid receptors, which take on the functional characteristics of AT1aR. The characterization of the chimera with stepwise replacement of the IC domains should allow for assignment of specific roles to the individual loops and C-terminus in the signaling and internalization of the BKB2R and facilitate the generation of a receptor with BKB2R binding and AT1aR function.


Adjunctive therapy with low molecular weight heparin with recombinant tissue-type plasminogen activator causes sustained reflow in canine coronary thrombosis.
Rethrombosis of the infarct-related artery after pharmacologic thrombolysis is a major limitation of the thrombolytic therapy. Platelet and fibrin deposition in the coronary artery after recombinant tissue-type plasminogen activator (rTPA) may play a leading role in reformation of thrombus. Therefore we examined the effect of low molecular weight heparin (LMWH) as adjunctive treatment with rTPA in a dog model of electrically induced intracoronary thrombus. Fourteen dogs, in which a stable coronary thrombus was induced with delivery of 100 microA of anodal direct current, were randomly given an intravenous bolus of LMWH, 75 IU/kg (n = 6), or saline (n = 8), followed by intravenous rTPA, 1 mg/kg over 20 minutes. LMWH (75 IU/kg) or saline was continuously infused over 90 minutes after rTPA-induced thrombolysis. Reperfusion occurred at 29 +/- 7 minutes in six of eight dogs receiving rTPA plus saline (reperfusion rate 75%), while reperfusion occurred at 18 +/- 3 minutes in all six dogs receiving rTPA plus LMWH (both p = NS versus rTPA plus saline group). Coronary reocclusion occurred in 83% of dogs given rTPA plus saline, but only in one dog (17%) given rTPA plus LMWH (p less than 0.05). Magnitude of reflow at 60 minutes of reperfusion was higher in the rTPA plus LMWH group than in the rTPA plus saline group (51 +/- 14 ml/min versus 10 +/- 9 ml/min; p less than 0.05). As expected, partial thromboplastin time was greater in rTPA plus LMWH than in rTPA plus saline-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS)


Gas chromatographic-mass spectrometric screening procedure for the identification of formaldehyde-derived tetrahydroisoquinolines in human urine.
A gas chromatographic-mass spectrometric method has been developed for the identification of 1,2,3,4-tetrahydroisoquinoline and six metabolites extracted from urine in the picogram range. The derivatization procedure for the substances, formed by reaction of formaldehyde with biogenic amines, employs propionic anhydride and can take place in aqueous medium. In this way artificial formation of these compounds via condensation of biogenic amines with aldehydes or alpha-keto acids during the work-up procedure is eliminated. The procedure results in hydrophobic compounds, which are quantitatively extractable by liquid-liquid extraction with organic solvents. Further clean-up was performed by solid-phase extraction on C18 sample preparation columns.


Surgical and oncological outcomes of an improved nerve-sparing radical hysterectomy technique: 6 years of experience at two centres.
An improved nerve-sparing radical hysterectomy (NSRH), which is based on the paravesico-vaginal space, has been recently introduced in a phase II, prospective clinical trial by our team. This study aims to report the surgical and oncological outcomes of this improved NSRH. One hundred seventy-seven consecutive patients were enrolled in our study and underwent the improved NSRH. The proportion of successful catheter removal and postvoid residual urine volume (PVR) of 50 mL or less at postoperative day 7 or day 4 was used to assess surgical outcomes. The local control rate (LCR), disease free survival (DFS), and overall survival (OS) were used to assess oncological outcomes. Postoperative 30-day complications occurred in 27/177 (15.3%) patients. The rate of successful catheter removal and PVR of 50 mL or less were 85.2% (23/27) and 66.7% (18/27) at postoperative day 7, and 73.3% (110/150) and 35.3% (53/150) at postoperative day 4. A total of 13 (7.9%) patients showed recurrence after a median follow-up time of 39.2 months (range 3.2-68.1 months). The estimated 2-year and 5-year DFS rates were 92.2% and 91.1%, respectively. Seven (4.2%) patients presented local recurrence, and five (3.0%) patients were dead at the end of the follow-up period. The estimated 5-year LCR and OS were 95.1% and 96.2%, respectively. In univariate analysis, International Federation of Gynecology and Obstetrics (FIGO) stage, lymphovascular space invasion (LVSI), and lymph node metastasis were found to be the prognostic risk factors of DFS. Patients with LVSI were associated with a worse DFS according to the multivariate analysis. The improved NSRH in our study may provide better surgical outcomes without compromising the survival in patients with early cervical cancer.


The predictive value of changes in left atrial volume index for rehospitalization in heart failure with preserved ejection fraction.
Left atrial volume index (LAVI) is an adequate analysis to predicate the left ventricle (LV) filling pressures, providing a powerful predictive marker of LV diastolic dysfunction. LAVI is a dynamic morphophysiological marker, and whether LAVI changes can predicate clinical outcomes in HF with preserved ejection fraction (HFpEF) is unknown. HFpEF patients were retrospectively studied from the First Affiliated Hospital of Dalian Medical University. Patients were classified into deteriorated, stable and improved groups according to the change in LAVI. Rehospitalization was defined as the main endpoint, the composite outcome of rehospitalization or all-cause death was defined as the secondary endpoint. A total of 409 patients were included. In this cohort, the percentage of deteriorated, stable, and improved LAVI were 99 (24.2%), 235 (57.4%), and 75 (18.4%), respectively. During the 22 months follow-up period, 168 patients (41.1%) were rehospitalized, 31 patients (7.5%) died and 182 patients (44.5%) experienced a composite outcome. Multivariate Cox regression showed that compared to improved LAVI, those with deteriorated and stable LAVI experienced higher risk of rehospitalization. Logistic regression showed atrial fibrillation (AF) and higher creatinine were independent predictors of deteriorated LAVI, whereas the use of loop diuretics, calcium channel blockers (CCB), and high level of high-density lipoprotein cholesterol (HDL-C) were significantly associated with improved LAVI. Change in LAVI provides a powerful and dynamic morphophysiological marker of LV filling status and can be used to evaluate the rehospitalization in HFpEF patients.


Anti-Fya-mediated delayed hemolytic transfusion reaction following emergency-release red blood cell transfusion: possible involvement of HLA-DRB1*04:03 in the Japanese population.
A 43-year-old Japanese male, who had undergone open liver surgery for tumor resection, presented with decreased hemoglobin levels on Day 13 post-emergency-release transfusion of 16 units of Fy(a +) red blood cells. As the anemia was accompanied by increased lactate dehydrogenase, indirect bilirubin, and reticulocytes, as well as decreased haptoglobin, it was attributed to hemolysis. In the diagnostic workup for hemolytic reaction, the direct antiglobulin test result for IgG was positive and the antibody dissociated from the patient's peripheral red blood cells was identified as anti-Fya (titer, 4). The hemolytic reaction was transient (approximately 10 days), of moderate severity, and did not result in any obvious organ damage. However, a single compatible red blood cell transfusion of 2 units was required on Day 17 after the causative transfusion. Notably, HLA typing revealed that the patient carried the HLA-DRB1*04:03 allele, which has been implicated in immunogenicity and induction of anti-Fya response in Caucasian populations. In summary, this is the first documented case of definitive anti-Fya-mediated delayed hemolytic transfusion reaction associated with HLA-DRB1*04:03 in the Japanese population.


Traumatic popliteal artery spasm diagnosed using intraoperative angiography: A case report.
There are few reports of traumatic arterial spasm in large-sized vessels in the extremities, and many surgeons are skeptical of its existence. We report a case of traumatic popliteal artery injury (PAI), which was later diagnosed as traumatic popliteal artery spasm on intraoperative angiography. A male patient, aged 54 years, was injured when a heavy object weighing approximately 100 kg fell on his right knee. Three hours after the injury, the patient was transported to our trauma center. The dorsalis pedis and posterior tibial arteries were not palpable, and his right lower leg was cold. However, he could move his ankle and toes completely and had no numbness or paresthesia in his right lower limb. Computed tomography angiography showed an interruption of the popliteal artery at the knee joint level and an enhancement on the distal side of the injury site. We diagnosed lower limb ischemia resulting from PAI and operated for revascularization. Under general anesthesia, the popliteal artery was explored via a posterior S-shaped incision in the prone position. No arterial abnormalities were observed macroscopically, and the injured area remained unidentified. Therefore, a contrast medium was injected into the popliteal artery using a 20-gauge intravascular needle, and angiography was performed, which revealed that the popliteal artery was patent; however, there was circumferential arterial stenosis at the level of the knee joint. At this point, the right lower limb's coldness had disappeared. Arterial spasm was speculated to cause the transient popliteal occlusion and lower leg ischemia. Immediate revascularization should be attempted when PAI is suspected. If no gross vascular abnormalities are confirmed, intraoperative angiography is useful for diagnosing arterial spasm.


Influence of growth phase on harvesting of Chlorella zofingiensis by dissolved air flotation.
The effects of changes in cellular characteristics and dissolved organic matter (DOM) on dissolved air flotation (DAF) harvesting of Chlorella zofingiensis at the different growth phases were studied. Harvesting efficiency increased with Al(3+) dosage and reached more than 90%, regardless of growth phases. In the absence of DOM, the ratio of Al(3+) dosage to surface functional group concentration determined the harvesting efficiency. DOM in the culture medium competed with algal cell surface functional groups for Al(3+), and more Al(3+) was required for cultures with DOM than for DOM-free cultures to achieve the same harvesting efficiency. As the culture aged, the increase of Al(3+) dosage due to increased DOM was less than the decrease of Al(3+) dosage associated with reduced cell surface functional groups, resulting in overall reduced demand for Al(3+). The interdependency of Al(3+) dosage and harvesting efficiency on concentrations of cell surface functional groups and DOM was successfully modeled.


Functional deacylases of pigeon liver fatty acid synthetase complex.
Fatty acid synthetase complex (Mr = 500,000) purified from pigeon liver homogenates is inactivated by phenylmethylsulfonyl fluoride. A well characterized inhibitor of serine esterases. Pseudounimolecular kinetics are followed at all inhibitor concentrations studied (0.05 to 1.0 mM). The second order rate constant obtained at pH 7.0, 30 degrees in 0.05 M potassium phosphate, 1 mM EDTA is 250 plus or minus 10 M-1 min-1 and appears to be independent of pH between 6 and 7.9. The inactivation of the enzyme complex appears to be selective since only one of the several component enzymes of fatty acid synthesis, palmityl-CoA deacylase, is inhibited. Acetyl- and malonyl-CoA-pantetheine transacylase activities as well as the kinetics of the reduction and dehydration steps are nearly identical for the native and the modified enzymes. The rate of approach of the condensation-CO2 exchange reaction (substrates: hexanoyl-CoA, malonyl-CoA, CoA, and H14CO3-) is slightly slower in the modified enzyme, though this change is not large enough to account for total loss of activity for fatty acid synthesis. The rate of loss of palmityl-CoA deacylase activity at a constant inhibitor concentration follows biphasic kinetics. Complete inactivation is achieved only after 2 mol of the inhibitor are bound per mol of the enzyme complex. Acetyl-, butyryl-, and hexanoyl-CoA thioesters (at 1.0 mM concentrations) protect the enzyme complex against inactivation by phenylmethylsulfonyl fluoride whereas CoA has no effect. Malonyl-CoA on the other hand, promotes inhibitor-mediated inactivation. Of the N-acetyl cysteamine derivatives tested, S-acetyl-N-acetyl cysteamine (at 10 mM) gives almost complete protection against inactivation whereas S-acetoacetyl-, S-beta-hydroxybutyryl-, and S-crotonyl-N-acetyl cysteamine thioesters exhibit either slight or no protection. These data demonstrate that phenylmethylsulfonyl fluoride is a selective reagent for the inactivation of functional fatty acyl deacylase component(s) of the pigeon liver fatty acid synthetase complex, and that it has no effect on malonyl or acetyl transacylases. The data are also in accord with the postulation that the inhibitor interacts at two catalytic centers of the enzyme complex. Furthermore, the patterns of protective effects shown by saturated acyl-CoA asters and malonyl-CoA point to different mechanisms of deacylation for these esters.


Non-negative matrix factorization model-based construction for molecular clustering and prognostic assessment of head and neck squamous carcinoma.
We aimed at exploring the efficacy of non-negative matrix factorization (NMF) model-based clustering for prognostic assessment of head and neck squamous carcinoma (HNSCC). The transcriptome microarray data of HNSCC samples were downloaded from The Cancer Genome Atlas (TCGA) and the Shanghai Ninth People's Hospital. R software packages were used to establish NMF clustering, from which relevant prognostic models were developed. Based on NMF, samples were allocated into 2 subgroups. Predictive models were constructed using differentially expressed genes between the two subgroups. The high-risk group was associated with poor prognostic outcomes. Moreover, multi-factor Cox regression analysis revealed that the predictive model was an independent prognostic predictor. The NMF-based prognostic model has the potential for prognostic assessment of HNSCC.


Cavernomas in children with brain tumors: a late complication of radiotherapy.
Delayed complications of radiation therapy comprise cerebral atrophy, radionecrosis and induction of tumors. Recent reports indicate the possibility of the "de-novo" formation of intracranial cavernomas in patients submitted to radiation therapy to the brain. To report three children, two with medulloblastomas and one with a pineal germinoma, treated with radiotherapy that developed intracerebral cavernous hemangiomas some years after treatment. With this work, we aim to draw attention to this occurrence in the neurosurgical community. The patients were two girls and one boy with ages comprised between 2.5 and 7 years (mean 5.2 years). The average interval from irradiation to the appearance of cavernoma was of 5.3 years (range 5-6 years). The lesions were found during the routine neuroimaging studies performed for the follow-up of their primary neoplasms. No patient showed signs or symptoms related to the cavernomas. However, the three children will need both clinical and neuroimaging surveillance to monitor the evolution of these incidentally discovered lesions. Intracranial cavernomas can occur years after cerebral radiation therapy. In spite of previous reports that show a high incidence of bleeding lesions, cavernomas may be found incidentally during the neuroimaging surveillance studies that are performed to children with brain tumors previously treated with radiotherapy. In these cases, a conservative attitude seems to be advisable, reserving surgery only for those lesions that grow or bleed.


Effect of Different Endurance Training Protocols During Cardiac Rehabilitation on Quality of Life.
This study aimed to assess the effect of different types of endurance training during outpatient cardiac rehabilitation on patients' health-related quality of life (HRQL). The MacNew Heart Disease HRQL questionnaire and the Hospital Anxiety and Depression Scale were used to assess changes in HRQL in 66 patients before and after 6 weeks of cardiac rehabilitation. Patients were randomized to 1 of 3 types of supervised endurance training: continuous endurance training, high-intensity interval training, and pyramid training. Two-way analysis of variance for repeated measure and chi-square test were used to analyze changes before and after rehabilitation. Attendance rate during the 6 weeks of exercise training was 99.2%. Physical work capacity increased from 136.1 to 165.5 watts (+22.9%; P < .001), and there were no statistical differences between training protocols. Fully completed questionnaires at both time points were available in 46 patients (73.9%; 61.3±11.6 years, 34 males, 12 females). Regardless of the type of supervised endurance training, there was significant improvement during rehabilitation in each of the categories of the MacNew questionnaire (ie, emotion, physical, social, global; all P < .05) and the Hospital Anxiety and Depression Scale (anxiety: P = .05; depression: P = .032), without significant differences between protocols. All 3 types of endurance training led to significant and well comparable increases in physical work capacity, which was associated with an increase in HRQL independent of the type of training. Our findings support further individualization of training regimes, which could possibly lead to better compliance during life-long home-based exercise training.


Polygenic Risk for Major Depression Interacts with Parental Criticism in Predicting Adolescent Depressive Symptom Development.
Research has focused more and more on the interplay between genetics and environment in predicting different forms of psychopathology, including depressive symptoms. While the polygenic nature of depressive symptoms is increasingly recognized, only few studies have applied a polygenic approach in gene-by-environment interaction (G × E) studies. Furthermore, longitudinal G × E studies on developmental psychopathological properties of depression are scarce. Therefore, this 6-year longitudinal community study examined the interaction between genetic risk for major depression and a multi-informant longitudinal index of critical parenting in relation to depressive symptom development from early to late adolescence. The sample consisted of 327 Dutch adolescents of European descent (56% boys; Mage T1 = 13.00, SDage T1 = 0.44). Polygenic risk for major depression was based on the Hyde et al. (Nature Genetics, 48, 1031-1036, 2016) meta-analysis and genetic sensitivity analyses were based on the 23andMe discovery dataset. Latent Growth Models suggested that polygenic risk score for major depression was associated with higher depressive symptoms across adolescence (significant main effect), particularly for those experiencing elevated levels of critical parenting (significant G × E). These findings highlight how polygenic risk for major depression in combination with a general environmental factor impacts depressive symptom development from early to late adolescence.