Direct effects of ciliary neurotrophic factor on brown adipocytes: evidence for a role in peripheral regulation of energy homeostasis.
Ciliary neurotrophic factor (CNTF) plays an important role in regulating neuronal growth. Recently, central anorexigenic effects of this cytokine have been characterized. However, peripheral effects on tissues that actively contribute to the regulation of energy homeostasis have not been described. Here, we report direct potent and selective effects of CNTF on growth factor and metabolic signalling intermediates in mouse brown adipocytes. CNTF stimulates STAT3, MAP kinase, Akt, and p70 S6 kinase. We find that, next to mediating Akt and p70 S6 kinase activation, both phosphatidylinositol 3-kinase and protein kinase C are separately acting, main intermediates for inducing mitogen-activated protein (MAP) kinase activation. On a functional level, CNTF enhances beta3-adrenergic induction of uncoupling protein-1. Thus, these results demonstrate direct effects of CNTF on adipose tissue signalling and metabolism and suggest a novel role for this cytokine in the peripheral regulation of energy homeostasis.


Safe Spaces: Mitigating Potential Aggression in Acute Care Psychiatry.
Awareness of potential aggression and violence is crucial when treating patients experiencing mental health crises in psychiatric emergency and inpatient settings. To provide a practical overview for health care workers in acute care psychiatry, the authors summarize relevant literature and clinical considerations on this important topic. Clinical contexts of violence in these settings, possible impact on patients and staff, and approaches to mitigating risk are reviewed. Considerations for early identification of at-risk patients and situations, and nonpharmacological and pharmacological interventions, are highlighted. The authors conclude with key points and future scholarly and practical directions that may further assist those entrusted with providing psychiatric care in these situations. Although working in these often high-paced, high-pressured settings can be challenging, effective violence-management strategies and tools can help staff optimize the focus on patient care while maintaining safety, their own well-being, and overall workplace satisfaction.


Radiological comparison of inferior turbinate hypertrophy between allergic and non-allergic rhinitis: does allergy really augment turbinate hypertrophy?
Inferior turbinate hypertrophy could be a result of allergic rhinitis (AR) that leads to nasal congestion and nasal airway obstruction, which is the most bothersome complaint in patients with AR. However, evidence regarding whether patients with AR have a more hypertrophied inferior turbinate than do patients with non-AR is lacking. We aimed to evaluate the degree of inferior turbinate hypertrophy according to the presence of AR using radiological measurements of the inferior turbinate. For evaluating the inferior turbinate, which contributes to nasal obstruction in patients with AR, we enrolled 90 adult patients with septal deviation and divided them into two groups (AR group: n = 49; non-AR group: n = 41). Allergic rhinitis was diagnosed according to the presence of an allergic history, positive multiple allergen simultaneous test, and serological total immunoglobulin E level (≥ 100 kU/L). We analyzed the minimal cross-sectional area on acoustic rhinometry for both groups. The bilateral total width as well as medial mucosa and nasal cavity space in the anterior and posterior portions of the inferior turbinate were measured using computed tomography. We could not find any significant differences in the anterior and posterior dimensions of the inferior turbinate, intranasal space, and choanal spaces between the AR and non-AR groups. Instead, the anterior part of the inferior turbinate in both the groups showed significant differences between the deviated and contralateral sides. The contralateral side had a larger width than did the deviated side, but no significant difference was noted in the posterior portion of the inferior turbinate. The degree of inferior turbinate hypertrophy showed no difference between patients with and without AR. Therefore, we suggest that surgical treatment for reducing the size of the inferior turbinate hypertrophy should be considered when performing septoplasty in patients with symptoms of nasal obstruction, regardless of the presence of AR.


Psychosocial factors influencing the disclosure of sexually transmissible infection diagnoses among female adolescents.
Individuals diagnosed with a sexually transmissible infection (STI) often face psychosocial concerns through which they must navigate to arrive at disclosure decisions. The objective of this study was to qualitatively explore the decision-making process for disclosure of STI diagnoses to sex partners among adolescent females in Baltimore City, MD. Semi-structured interviews were conducted with 21 African American adolescent females who tested positive for chlamydia and/or gonorrhoea in the previous 3 months. Three individuals also provided audio diaries in which they tape recorded a daily journal discussing their lives and experiences with STIs. Female adolescents often do not notify all potentially infected sex partners or tell sex partners all of the STIs to which they may have been exposed. Although participants recognised the importance of informing their sex partner(s) of their STI diagnosis, there were other competing psychosocial factors that influenced their decisions to disclose, such as perceptions of the social consequences of having an STI including fear of stigmatisation and loss of relationship. Such concerns appear to vary by disease and partner type as well as previous STI experience. To increase disclosure of STI diagnoses, clinic counselling strategies to encourage partner notification should seek to engage patients in a dialogue regarding their fears related to their diagnoses, how this might affect their relationships with different types of partners and the individual and social meanings their specific STI diagnosis(es) might hold.


Respiration, Heartbeat, and Conscious Tactile Perception.
Previous studies have shown that timing of sensory stimulation during the cardiac cycle interacts with perception. Given the natural coupling of respiration and cardiac activity, we investigated here their joint effects on tactile perception. Forty-one healthy female and male human participants reported conscious perception of finger near-threshold electrical pulses (33% null trials) and decision confidence while electrocardiography, respiratory activity, and finger photoplethysmography were recorded. Participants adapted their respiratory cycle to expected stimulus onsets to preferentially occur during late inspiration/early expiration. This closely matched heart rate variation (sinus arrhythmia) across the respiratory cycle such that most frequent stimulation onsets occurred during the period of highest heart rate probably indicating highest alertness and cortical excitability. Tactile detection rate was highest during the first quadrant after expiration onset. Interindividually, stronger respiratory phase-locking to the task was associated with higher detection rates. Regarding the cardiac cycle, we confirmed previous findings that tactile detection rate was higher during diastole than systole and newly specified its minimum at 250-300 ms after the R-peak corresponding to the pulse wave arrival in the finger. Expectation of stimulation induced a transient heart deceleration which was more pronounced for unconfident decision ratings. Interindividually, stronger poststimulus modulations of heart rate were linked to higher detection rates. In summary, we demonstrate how tuning to the respiratory cycle and integration of respiratory-cardiac signals are used to optimize performance of a tactile detection task.SIGNIFICANCE STATEMENT Mechanistic studies on perception and cognition tend to focus on the brain neglecting contributions of the body. Here, we investigated how respiration and heartbeat influence tactile perception: respiration phase-locking to expected stimulus onsets corresponds to highest heart rate (and presumably alertness/cortical excitability) and correlates with detection performance. Tactile detection varies across the heart cycle with a minimum when the pulse reaches the finger and a maximum in diastole. Taken together with our previous finding of unchanged early event-related potentials across the cardiac cycle, we conclude that these effects are not a peripheral physiological artifact but a result of cognitive processes that model our body's internal state, make predictions to guide behavior, and might also tune respiration to serve the task.


Investigating changes in mental health services utilisation in England and their impact on health outcomes and wellbeing during the COVID-19 pandemic: Protocol for a health data-linkage study.
Linking routinely collected health care system data records for the same individual across different services and over time has enormous potential for the NHS and its patients. The aims of this data linkage study are to quantify the changes to mental health services utilisation in responses to the COVID-19 pandemic and determine whether these changes were associated with health-related outcomes and wellbeing among people living in the most deprived communities in North East and North Cumbria, England. We will assemble a retrospective cohort of people having referred or self-referred to NHS-funded mental health services or Improving Access to Psychological Therapies (IAPT) services between 23rd March 2019 and 22nd March 2020 in the most deprived areas in England. We will link together data from retrospective routinely collected healthcare data including local general practitioner (GP) practice data, Hospital Episode Statistics admitted patient care outpatients, and A&E, Community Services Data Set, Mental Health Services Data Set, and Improving Access to Psychological Therapies Data Set. We will use these linked patient-level data to 1) describe the characteristics of the cohort prior to the lockdown; 2) investigate changes to mental health services utilised between multiple time periods of the COVID-19 lockdown including out of lockdown; 3) explore the relationship between these changes and health outcomes/wellbeing and factors that confound and mediate this relationship among this cohort. This study comprises a deprived population-based cohort of people having referred or self-referred to NHS-funded secondary mental health services or Improving Access to Psychological Therapies (IAPT) services over an extended period of the lockdown in England (2019-2022).This study will utilise a new longitudinal data resource that will link together detailed data from a cohort of individual participants and retrospective administrative data relating to the use of primary, secondary, and community care services.The study period covers pre-lockdown, different lockdown and post-lockdown, and out of lockdown periods up to March 2022.Routinely collected administrative data contain limited contextual information and represent an underestimate of total health outcomes for these individuals.Routinely collected datasets can often been incomplete or contain missing data, which can make it difficult to accurately analyse the data and draw meaningful conclusions.Intervention and treatment for mental health conditions are not wholly captured across these data sources and may impact health outcomes.


Central neurocytoma: morphological, flow cytometric, polymerase chain reaction, fluorescence in situ hybridization, and karyotypic analyses. Case report.
The results of cytogenetic and molecular genetic analysis of a central neurocytoma are presented. Central neurocytomas are intriguing neoplasms that exhibit primarily neuronal, but also glial characteristics, which indicate an origin from a pluripotential neuroglial precursor. The authors describe an intraventricular neurocytoma in an 11-year-old boy that showed anaplastic features with widespread necrosis and mitoses, as well as extensive calcification and foci that exhibited marked neuronal differentiation with clusters of ganglion cells. Immunohistochemical examination showed prominent synaptophysin and neurofilament positivity and focal glial fibrillary acidic protein positivity. Electron microscopy revealed abundant neuritic processes with microtubules and dense core granules as well as mature ganglion cells. Flow cytometry studies revealed increased S (7.8%) and G2M (9.7%) phase components. Molecular and cytogenetic studies were undertaken to assess whether there were similarities to two other tumor types that exhibit neuronal differentiation, the neuroblastoma and medulloblastoma. Polymerase chain reaction and fluorescence in situ hybridization (FISH) analysis revealed no evidence of amplification of the MYCN oncogene or chromosome 1p deletion, which are common in neuroblastomas. Chromosomal analysis by G banding revealed a complex karyotype, with counts in the near-diploidy range (45-48). Two chromosomes 1 appeared normal on G banding and FISH analysis, with p58 signals present on the distal p arm of both chromosomes 1; however, three additional copies of distal 1q were present in rearrangements with 4 and 7. Although the histological findings indicate a kinship to the neuroblastoma and medulloblastoma, the central neurocytoma appears to have a different karyotypic profile, although more cases need to be assessed using molecular genetic analysis.


Deep Boosted Molecular Dynamics: Accelerating Molecular Simulations with Gaussian Boost Potentials Generated Using Probabilistic Bayesian Deep Neural Network.
We have developed a new deep boosted molecular dynamics (DBMD) method. Probabilistic Bayesian neural network models were implemented to construct boost potentials that exhibit Gaussian distribution with minimized anharmonicity, thereby allowing for accurate energetic reweighting and enhanced sampling of molecular simulations. DBMD was demonstrated on model systems of alanine dipeptide and the fast-folding protein and RNA structures. For alanine dipeptide, 30 ns DBMD simulations captured up to 83-125 times more backbone dihedral transitions than 1 μs conventional molecular dynamics (cMD) simulations and were able to accurately reproduce the original free energy profiles. Moreover, DBMD sampled multiple folding and unfolding events within 300 ns simulations of the chignolin model protein and identified low-energy conformational states comparable to previous simulation findings. Finally, DBMD captured a general folding pathway of three hairpin RNAs with the GCAA, GAAA, and UUCG tetraloops. Based on a deep learning neural network, DBMD provides a powerful and generally applicable approach to boosting biomolecular simulations. DBMD is available with open source in OpenMM at https://github.com/MiaoLab20/DBMD/.


Prevention of relapses with levamisole as adjuvant therapy in children with a first episode of idiopathic nephrotic syndrome: study protocol for a double blind, randomised placebo-controlled trial (the LEARNS study).
Idiopathic nephrotic syndrome (INS) is characterised by a high relapse rate up to 80% after initial response to standard therapy with corticosteroids. Steroid toxicity is common and causes a great burden of disease that negatively influences the health-related quality of life (HRQoL). Recently, studies have shown that levamisole, an anthelminthic drug, significantly improves relapse-free survival in children with frequent relapses or steroid dependency. Compared with other steroid-sparing drugs, levamisole has relatively few side effects. We hypothesise that adding levamisole to standard therapy with corticosteroids in children with a first episode of INS will prevent relapses, decrease cumulative dosage of steroids used and improve HRQoL. This paper presents the study protocol for the LEARNS study (LEvamisole as Adjuvant therapy to Reduce relapses of Nephrotic Syndrome). An international, double-blind, placebo-controlled randomised trial will be conducted in 20 participating hospitals in the Netherlands and Belgium. Participants (n=92) with a first episode of INS, aged 2-16 years, who achieve remission after 4 weeks of oral prednisolone will be randomly assigned (1:1) to receive either levamisole 2.5 mg/kg alternate day or placebo added to prednisolone (18-week tapering schedule) for a total of 24 weeks. Follow-up will be until 2 years after first presentation. Additionally, parents and/or children will fill out five HRQoL questionnaires. Primary outcome of the LEARNS study is occurrence of relapses within 12 months after first presentation. Secondary outcomes include time to first relapse, cumulative steroid dose after 2 years, safety parameters and quality of life scores. The trial was approved by the Medical Ethical Committee. Results of the study will be published in a peer-reviewed journal. NL6826, 2017-001025-41.


Longitudinal noninvasive imaging of progesterone receptor as a predictive biomarker of tumor responsiveness to estrogen deprivation therapy.
To investigate whether longitudinal functional PET imaging of mammary tumors using the radiopharmaceuticals [(18)F]FDG (to measure glucose uptake), [(18)F]FES [to measure estrogen receptor (ER) levels], or [(18)F]FFNP [to measure progesterone receptor (PgR) levels] is predictive of response to estrogen-deprivation therapy. [(18)F]FDG, [(18)F]FES, and [(18)F]FFNP uptake in endocrine-sensitive and -resistant mammary tumors was quantified serially by PET before ovariectomy or estrogen withdrawal in mice, and on days 3 and 4 after estrogen-deprivation therapy. Specificity of [(18)F]FFNP uptake in ERα(+) mammary tumors was determined by competition assay using unlabeled ligands for PgR or glucocorticoid receptor (GR). PgR expression was also assayed by immunohistochemistry (IHC). The levels of [(18)F]FES and [(18)F]FDG tumor uptake remained unchanged in endocrine-sensitive tumors after estrogen-deprivation therapy compared with those at pretreatment. In contrast, estrogen-deprivation therapy led to a reduction in PgR expression and [(18)F]FFNP uptake in endocrine-sensitive tumors, but not in endocrine-resistant tumors, as early as 3 days after treatment; the changes in PgR levels were confirmed by IHC. Unlabeled PgR ligand R5020 but not GR ligand dexamethasone blocked [(18)F]FFNP tumor uptake, indicating that [(18)F]FFNP bound specifically to PgR. Therefore, a reduction in FFNP tumor to muscle ratio in mammary tumors predicts sensitivity to estrogen-deprivation therapy. Monitoring the acute changes in ERα activity by measuring [(18)F]FFNP uptake in mammary tumors predicts tumor response to estrogen-deprivation therapy. Longitudinal noninvasive PET imaging using [(18)F]FFNP is a robust and effective approach to predict tumor responsiveness to endocrine treatment.


[Residue situation of milk products after the effective carrying out of the "step-by-step program for DDT-replacement" in the GDR].
A step-by-step program had been established to replace the persistent insecticide DDT with other active principles in the GDR by 1975. Monthly thin-layer chromatographic residue determinations in butter samples from all the districts demonstrate that the DDT-residues have decreased gradually since 1973 from an average value of 0.45 mg/kg to an average value of 0.18 mg/kg (in 1976). The DDT intake of adults via the milk products (butter, milk and cheese) was, in 1976, only 2% of the ADI value, which evidences a very positive development. Furthermore, findings concerning the contamination with lindane and HCB are presented.


Management of combined sewer overflows based on observations from the urbanized Liguori catchment of Cosenza, Italy.
This paper examines an urbanized catchment in Cosenza, Italy where an off-line basin intercepting CSOs was studied to illustrate reduction in CSO discharges to the Crati River. While the hydrologic transport of pollutant mass is never known a-priori and can be flow-limited, the volumetric requirements of the basin were modeled based on the classic assumption that wet weather flows transport urban and sewer loads in a mass-limited (first-flush) delivery. The volumetric capacity of the basin was varied from 10 to 50 m(3)/ha. Operational basin control was simulated with historical datasets from the Liguori catchment, event-based loading data, and continuous simulation modelling with SWMM. Utilizing data from the catchment, the SWMM simulations were conducted considering the storage basin with or without sedimentation treatment. Results illustrate the potential benefits of the off-line operation for the system with respect to the volume and mass reduction of CSOs into the Crati River. Results demonstrate the importance of particle size distribution (PSD) as an index of basin efficiency, coupled with analysis of the hydrodynamic response of the basin. The basin model attenuated influent PSDs, separating the coarser fraction of the PSD, and reduced the load of influent particulate matter (PM).


8-Aminocaprylic acid.
The title acid, 8-aminooctanoic acid, C8H17NO2, crystallized in the centrosymmetric space group P2(1)/n in the zwitterionic form. The three H atoms involved in hydrogen bonding are ordered. The five intermolecular N-H...O hydrogen bonds have N...O distances ranging from 2.752 (2) to 3.258 (2) A and N-H...O angles ranging from 131 (2) to 165 (2) degrees. Each molecule is linked to six neighboring molecules by a total of ten hydrogen bonds. A complex network of hydrogen bonds ensues in which chains predominate.


Failure of pulse high-dose dexamethasone in chronic idiopathic immune thrombocytopenia.
Idiopathic thrombocytopenic purpura (ITP) is a disorder characterized by increased platelet destruction in the setting of normal megakaryopoiesis. Approximately 20% of patients with ITP are refractory to corticosteroids and splenectomy. Recently, pulse high-dose dexamethasone was reported to be effective in the treatment of chronic ITP in adult patients. We treated 9 patients with severe chronic ITP with monthly high-dose dexamethasone. None of the 9 patients responded with a sustained increase in platelet count. Five of these patients were unable to tolerate the regimen. The failure of high-dose dexamethasone in our hands contrasts with the good results of an earlier publication and suggests that there could be a subset of responders who will require better identification.


Survalytics: An Open-Source Cloud-Integrated Experience Sampling, Survey, and Analytics and Metadata Collection Module for Android Operating System Apps.
We describe here Survalytics, a software module designed to address two broad areas of need. The first area is in the domain of surveys and app analytics: developers of mobile apps in both academic and commercial environments require information about their users, as well as how the apps are being used, to understand who their users are and how to optimally approach app development. The second area of need is in the field of ecological momentary assessment, also referred to as experience sampling: researchers in a wide variety of fields, spanning from the social sciences to psychology to clinical medicine, would like to be able to capture daily or even more frequent data from research subjects while in their natural environment. Survalytics is an open-source solution for the collection of survey responses as well as arbitrary analytic metadata from users of Android operating system apps. Surveys may be administered in any combination of one-time questions and ongoing questions. The module may be deployed as a stand-alone app for experience sampling purposes or as an add-on to existing apps. The module takes advantage of free-tier NoSQL cloud database management offered by the Amazon Web Services DynamoDB platform to package a secure, flexible, extensible data collection module. DynamoDB is capable of Health Insurance Portability and Accountability Act compliant storage of personal health information. The provided example app may be used without modification for a basic experience sampling project, and we provide example questions for daily collection of blood glucose data from study subjects. The module will help researchers in a wide variety of fields rapidly develop tailor-made Android apps for a variety of data collection purposes.


Androgen receptor signaling fuels DNA repair and radioresistance in prostate cancer.
Successful treatment by genotoxic modalities including radiotherapy is commonly hampered by treatment resistance in advanced cancers. Two new studies now reveal that androgen receptor signaling transcriptionally upregulates a large subset of DNA repair genes, thereby enhancing the repair capacity and promoting radioresistance of prostate cancer. These results provide a mechanistic rationale for a combined treatment by ionizing radiation and androgen deprivation therapy.


Fatal fatty liver-kidney syndrome in obese monkeys.
A fatal syndrome associated with brief periods of anorexia or acute weight loss was noted in monkeys. Affected monkeys died unexpectedly or after a very short illness. Consistent gross findings at necropsy were enlarged, pale yellow livers, pale tan to yellow kidneys, abundant deposits of body fat and empty gastrointestinal tracts. Fatty change of the liver and kidney was the predominant and characteristic lesion found microscopically. Most of the monkeys were female Macaca fascicularis greater than 8 years of age. However, monkeys of other species and ages, and one male also died of this syndrome. Affected monkeys generally had losses of 8-33% of body weight (last measured weight--weight at death). In some animals, this syndrome was associated with a location change forcing new social interactions. A common clinical pathologic finding was azotemia. These findings suggest that obese monkeys may be prone to a fatal syndrome characterized by fatty change of the liver and kidney, and weight loss.


Does death anxiety affect end-of-life care discussions?
The aim of this study was to determine if a gynecologic cancer patient's comfort level discussing end-of-life care issues with her caregivers is related to her death anxiety level. Gynecologic oncology clinic patients were asked to rate their degree of agreeability with 4 statements regarding comfort level discussing end-of-life care issues. Participants also completed the Hoge's Intrinsic Religiosity Scale and Templer's Death Anxiety Scale. Four hundred one surveys were distributed. One hundred twenty-nine patients participated, with a response rate of 32.2%. The median age of the sample was 55 years. Most patients were white (72.9%), married (58.9%), and Christian (85.3%). Most patients had ovarian cancer (40.4%). Of the 74 patients who knew their cancer stage, 59% had been diagnosed with advanced (stage III-IV) disease. Thirty-three percent were currently in remission, and 17% had recurrent disease. Of all patients surveyed, 32.6% were currently receiving treatment. Chemotherapy was the most common treatment (62% of those being treated). Higher level of comfort discussing end-of-life care topics such as do-not-resuscitate orders with family members was significantly associated with decreased death anxiety (P = 0.008 and P = 0.001). There was no significant association between comfort level when patients discussed do-not-resuscitate orders with physicians and patients' death anxiety (P = 0.14). After controlling for age, race, marital status, education level, current treatment status, and religiosity, linear regression analysis demonstrated that the relationship between a patient's increased comfort level when discussing end-of-life care topics with family members and decreased death anxiety remained statistically significant (P = 0.005 and P = 0.001). Conversations regarding goals of treatment are an important component of caring for cancer patients. Death anxiety may contribute to decreased communication between patients and their family members regarding the patient's end-of-life care wishes. Obtaining a better understanding of the role death anxiety plays in end-of-life care discussions may help patients receive the end-of-life care they desire.


Skeletal muscle structure and function in response to electrical stimulation in moderately impaired COPD patients.
To determine the structural and functional consequences of high-frequency neuromuscular electrical stimulation (hf-NMES) in a group of moderately impaired outpatients with chronic obstructive pulmonary disease (COPD). A prospective, cross-over randomized trial. An university-based, tertiary center. Seventeen patients (FEV(1)=49.6+/-13.4% predicted, Medical Research Council dyspnoea grades II-III) underwent 6-weeks hf-NMES (50Hz) and sham stimulation of the quadriceps femoris in a randomized, cross-over design. Knee strength was measured by isokinetic dynamometry (peak torque) and leg muscle mass (LMM) by DEXA; in addition, median cross-sectional area (CSA) of type I and II fibres and capillary-fibre ratio were evaluated in the vastus lateralis. The 6-min walking distance (6MWD) was also determined. At baseline, patients presented with well-preserved functional capacity, muscle strength and mass: there was a significant relationship between strength and type II CSA (P<0.05). NMES was not associated with significant changes in peak torque, LMM or 6MWD as compared to sham (P>0.05). At micro-structural level, however, electrical stimulation increased type II, but decreased type I, CSA; no change, however, was found in the relative fibre distribution or capillary:fibre ratio (P<0.05). There was no significant association between individual changes in structure and function with training (P>0.05). Post-NMES increase in type II CSA was inversely related to baseline mass and strength (P<0.05). NMES may promote a modest degree of type II muscle fibre hypertrophy in COPD patients with well-preserved functional status. These micro-structural changes, however, were not translated into increased volitional strength in this sub-population.


Never-frozen liquid plasma transfusion in civilian trauma: a nationwide propensity-matched analysis.
Never-frozen liquid plasma (LQP) was found to reduce component waste, decrease health care expenses, and have a superior hemostatic profile compared with fresh frozen plasma (FFP). Although transfusing LQP in hemorrhaging patients has become more common, its clinical effectiveness remains to be explored. This study aims to examine outcomes of trauma patients transfused with LQP compared with thawed FFP. Adult (≥18 years) trauma patients receiving early (≤4 hours) plasma transfusions were identified in the Trauma Quality Improvement Program 2017. Patients were stratified into those receiving LQP versus FFP. Propensity-score matching in a 1:2 ratio was performed. Primary outcome measures were mortality and time to first plasma unit transfusion. Secondary outcome measures were major complications and hospital length of stay. A total of 107 adult trauma patients receiving LQP were matched to 214 patients receiving FFP. Mean age was 48 ± 19 years, 73% were male, and median Injury Severity Score was 27 [23-41]. A total of 42% of patients were in shock, 22% had penetrating injuries, and 31% required surgical intervention for hemorrhage control. Patients received a median of 4 [2-6] units of PRBC, 2 [1,3] units of LQP or FFP, and 1 [0-1] unit of platelets. The median time to the first LQP unit transfused was significantly shorter compared with the first FFP unit transfused (54 [28-79] minutes vs. 98 [59-133] minutes; p < 0.001). Rates of 24-hour mortality (2.8% vs. 3.7%; p = 0.664) and in-hospital mortality (16.8% vs. 20.1%; p = 0.481) were not different between the LQP and FFP groups. Similarly, there was no difference in major complications (15.9% vs. 21.5%; p = 0.233) and hospital length of stay (12 [6-21] vs. 12 [6-23] days; p = 0.826). Never-frozen liquid plasma is safe and effective in resuscitating trauma patients. Never-frozen liquid plasma has the potential to expand our transfusion armamentarium given its longer storage time and immediate availability. Therapeutic, Level IV.


A Diagnostic Model to Improve the Predictability of Natural Pregnancy Potential in Patients with Oligoasthenospermia.
BACKGROUND Oligoasthenospermia is one of the major reasons for male infertility in clinical practice. Nevertheless, some patients with oligoasthenospermia show normal fertility. Currently, there is a lack of an effective method to distinguish patients with oligoasthenospermia showing normal fertility from those who lack natural fertility and should participate in in vitro fertilization and assisted reproduction. MATERIAL AND METHODS In this study, we collected semen and blood samples from 153 males of Shui nationality at reproductive age in Guizhou Province, southwest China. We measured the routine parameters for semen and some serological indicators. A clinical diagnosis model was then constructed to evaluate the fertility potential of oligoasthenospermia patients using a logistic stepwise regression method, which was then visualized with a nomogram. RESULTS Our results showed that this model could effectively assess the natural pregnancy potential of patients with oligoasthenospermia, and its sensitivity and specificity were superior to those of a traditional model that used only sperm motility and count to assess male fertility potential (area under the curve=0.7626 vs. 0.6677). Additionally, we evaluated the clinical net benefit for patients with oligoasthenospermia at different risk scores in our model using decision curve analysis. The results showed that the net benefit was obtained at scores ranging from 0.1 to 0.6. CONCLUSIONS This comprehensive clinical prediction model can be used to determine whether infertile oligoasthenospermia patients lack natural fertility.


Prevalence and Amounts of Common Ingredients Found in Energy Drinks and Shots.
Energy drinks are one of the most popular packaged beverage products consumed within the United States (US). Energy drinks are considered a functional beverage, a category that also includes sports drinks and nutraceutical beverages. The focus of the current study was to examine the nutrition fact panels of the top selling commercially available energy drink and energy shot products within the US to characterize common ingredient profiles to help establish a standard definition and ingredient profile of energy drinks and energy shots for consumers, health care practitioners, and researchers. The top 75 commercially available energy drinks and shots were identified and compiled from multiple commercial retail websites as of September 2021. For the purpose of this study, an energy drink must have met the following criteria: (A) marketed as an energy drink; (B) purported to improve energy, focus, or alertness; (C) not sold as a dietary supplement (no supplement fact panels); (D) manufactured as a pre-packaged and ready-to-drink beverage; and (E) contains at least three of (1) caffeine, (2) B-vitamins, (3) sugar, (4) taurine, (5) creatine, (6) quercetin, (7) guarana, (8) ginseng, (9) coenzyme Q10, or (10) branched chain amino acids. Energy shots must have met similar criteria to be included: (A) marketed as an energy shot; (B) purported to improve energy, focus, or alertness; (C) sold as a dietary supplement; (D) manufactured as a pre-packaged beverage with a small volume (<3.5 mL); and (E) contains at least three of the ingredients stated above. Twenty energy shots and fifty-five energy drinks were included in this analysis. The number of ingredients per product (mean ± SD) was 18.2 ± 5.7, with 15 products containing proprietary blends with undisclosed ingredient amounts. The relative prevalence and average amounts of the top ingredients were as follows: caffeine (100%; 174.4 ± 81.1 mg), vitamin B6 (72%; 366.9 ± 648.1 percent daily value (%DV)), vitamin B3 (67%; 121.44 ± 69.9% DV), vitamin B12 (67%; 5244.5 ± 10,474.6% DV), vitamin B5 (37.3%; 113.6 ± 76.6% DV), and taurine (37.3%; amounts undisclosed). Our findings suggest a high prevalence of caffeine and B-vitamins in these energy products, with many of the formulations containing well above the recommended daily value of B-vitamins.


[High performance liquid chromatography in studies of component composition of polyene macrolides].
The published data on the use of HPLC in studies on the chemistry of polyene macrolide antibiotics are summarized and the conditions for HPLC, the use of various sorbents, the impact of the ratio of the organic solvents and buffer solutions, etc. are discussed comparatively. Several aspects of the use of HPLC are indicated i.e. the identification of polyene antibiotics (heptaenes, pentaenes and hexaenes), the standardization and investigation of the antibiotic homogeneity and quality, the purification of the antibiotics and the determination of the amphotericin B content in tissues and plasma.


Ultrasound image processing to estimate the structural and functional properties of mouse skeletal muscle.
Noninvasive imaging techniques are increasingly used for monitoring muscle behavior in mice. However, muscle is a complex tissue that exhibits different properties under passive and active conditions. In addition to structural properties, it is also important to analyze functional characteristics. At present, such information can be obtained with ultrasound elastography. However, this technique is poorly used for small rodent models (mice and gerbils). Thus, this study aims at establish referent hindlimb muscle data, and experimental guidelines, for wild-type (WT) control mice as well as the TIEG1 knockout (KO) mouse model that is known to exhibit skeletal muscle defects. Ultrasound was performed with the Aixplorer machine using a SLH 20-6 linear transducer probe (2.8 cm footprint). A region of interest (ROI) was placed around a superficial group of muscles. Subsequently, from the B-mode image, a classification of all the muscles and ultrasound biomarkers such as echo intensity and texture anisotropy have been determined. The influence of the gain setting (from 40% to 70%) was analyzed on these parameters. Moreover, the elasticity (E) was also measured within the ROI. This study provides a suitable methodology for collecting experimental data: 1) the correct range of gain (between 50% and 70%) to apply for the ultrasound measurement of muscle structure, 2) the structural and functional referent data for a group of healthy muscles, 3) the gray scale index, the texture anisotropy and the elasticity (ETIEG1 KO = 36.1 ± 10.3 kPa, EWT = 44.4 ± 13.4 kPa) parameters, which were obtained for a group of muscles as a function of genotype.


Repurposing of Biologic and Targeted Synthetic Anti-Rheumatic Drugs in COVID-19 and Hyper-Inflammation: A Comprehensive Review of Available and Emerging Evidence at the Peak of the Pandemic.
Coronavirus disease 2019 (COVID-19) is a condition caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Severe cases of COVID-19 result in acute respiratory distress syndrome and death. A detrimental, hyper-inflammatory immune response with excess release of cytokines is the main driver of disease development and of tissue damage in these patients. Thus, repurposing of biologic agents and other pharmacological inhibitors of cytokines used for the treatment of various inflammatory conditions emerged as a logical therapeutic strategy to quench inflammation and improve the clinical outcome of COVID-19 patients. Evaluated agents include the interleukin one receptor blocker anakinra, monoclonal antibodies inhibiting IL-6 tocilizumab and sarilumab, monoclonal antibodies inhibiting granulocyte-monocyte colony stimulating factor and tumor necrosis factor, and Janus kinase inhibitors. In this review, we discuss the efficacy and safety of these therapeutic options based on direct personal experience and on published evidence from observational studies and randomized clinical trials.


Facile Multicomponent Polymerizations toward Unconventional Luminescent Polymers with Readily Openable Small Heterocycles.
Heterocyclic polymers have gained enormous attention for their unique functionalities and wide applications. In contrast with the well-studied polymer systems with five- or six-membered heterocycles, functional polymers with readily openable small-ring heterocycles have rarely been explored due to their large synthetic difficulty. Herein, a facile one-pot multicomponent polymerization to such polymers is developed. A series of functional polymers with multisubstituted and heteroatom-rich azetidine frameworks are efficiently generated at room temperature in high atom economy from handy monomers. The four-membered azetidine rings in the polymer skeletons can be easily transformed into amide and amidine moieties via a fast and efficient acid-mediated ring-opening reaction, producing brand-new polymeric materials with distinctive properties. All the as-prepared azetidine-containing polymers exhibit intrinsic visible luminescence in the solid state under long-wavelength UV irradiation even without conventionally conjugated structures. Such unconventional luminescence is attributed to the clusteroluminogens formed by through-space electronic interactions of heteroatoms and phenyl rings. All the obtained polymers show excellent optical transparency, high and tunable refractive indices, low optical dispersions and good photopatternability, which make them promising materials in various advanced electronic and optoelectronic devices. The ring-opened polymers can also function as a lysosome-specific fluorescent probe in biological imaging.


Identification and development of a functional marker of TaGW2 associated with grain weight in bread wheat (Triticum aestivum L.).
The OsGW2 gene is involved in rice grain development, influencing grain width and weight. Its ortholog in wheat, TaGW2, was considered as a candidate gene related to grain development. We found that TaGW2 is constitutively expressed, with three orthologs expressing simultaneously. The coding sequence (CDS) of TaGW2 is 1,275 bp encoding a protein with 424 amino acids, and has a functional domain shared with OsGW2. No divergence was detected within the CDS sequences in the same locus in ten varieties. Genome-specific primers were designed based on the sequence divergence of the promoter regions in the three orthologous genes, and TaGW2 was located in homologous group 6 chromosomes through CS nulli-tetrasomic (NT). Two SNPs were detected in the promoter region of TaGW2-6A, forming two haplotypes: Hap-6A-A (-593A and -739G) and Hap-6A-G (-593G and -769A). A cleaved amplified polymorphic sequence (CAPS) marker was developed based on the -593 A-G polymorphism to distinguish the two haplotypes in TaGW2-6A. This gene was fine mapped 0.6 cM from marker cfd80.2 near the centromere in a recombinant inbred line (RIL) population. Two hundred sixty-five Chinese wheat varieties were genotyped and association analysis revealed that Hap-6A-A was significantly associated with wider grains and higher one-thousand grain weight (TGW) in two crop seasons. qRT-PCR revealed a negative relationship between TaGW2 expression level and grain width. The Hap-6A-A frequencies in Chinese varieties released at different periods showed that it had been strongly positively selected in breeding. In landraces, Hap-6A-A is mainly distributed in southern Chinese wheat regions. Association analysis also indicated that Hap-6A-A not only increased TGW by more than 3 g, but also had earlier heading and maturity. In contrast to Chinese varieties, Hap-6A-G was the predominant haplotype in European varieties; Hap-6A-A was mainly present in varieties released in the former Yugoslavia, Italy, Bulgaria, Hungary and Portugal.


Antigenicity of the Mu (B.1.621) and A.2.5 SARS-CoV-2 Spikes.
The rapid emergence of SARS-CoV-2 variants is fueling the recent waves of the COVID-19 pandemic. Here, we assessed ACE2 binding and antigenicity of Mu (B.1.621) and A.2.5 Spikes. Both these variants carry some mutations shared by other emerging variants. Some of the pivotal mutations such as N501Y and E484K in the receptor-binding domain (RBD) detected in B.1.1.7 (Alpha), B.1.351 (Beta) and P.1 (Gamma) are now present within the Mu variant. Similarly, the L452R mutation of B.1.617.2 (Delta) variant is present in A.2.5. In this study, we observed that these Spike variants bound better to the ACE2 receptor in a temperature-dependent manner. Pseudoviral particles bearing the Spike of Mu were similarly neutralized by plasma from vaccinated individuals than those carrying the Beta (B.1.351) and Delta (B.1.617.2) Spikes. Altogether, our results indicate the importance of measuring critical parameters such as ACE2 interaction, plasma recognition and neutralization ability of each emerging variant.


Characterization of MCF-12A cell phenotype, response to estrogens, and growth in 3D.
Three-dimensional cultures of mammary epithelial cells allow for biologically-relevant studies of the development of the mammary gland in rodents and humans under normal and pathological conditions, like carcinogenesis. Under these conditions, mammotropic hormones play significant roles in tissue morphogenesis. Therefore, a system that recreates the normal, hormonally responsive epithelium would be a valuable tool to study the normal state and its transition to carcinogenesis. MCF-12A cells have been claimed to be non-tumorigenic mammary epithelial cells with reported sensitivity to estrogens. In this study, we aimed at characterizing MCF-12A cells for use in a hormone-responsive 3D culture system to determine their usefulness as a tool to identify normal and abnormal microenvironmental cues. MCF-12A cells were single-cell cloned in order to investigate their heterogeneous makeup. The parental cells were then treated with estradiol to investigate proliferative and transcriptional responses through the estrogen receptor alpha. Finally, parental cells and epithelial-like cell-derived clones were seeded in rat-tail collagen I to profile the morphogenesis of multicellular 3D structures. The resultant structures were then analyzed using unsupervised morphometric analysis. MCF-12A cells consist of epithelial-like colonies which shed elongated, freely growing cells on the colony's edges. The cells express E-cadherin as well as mesenchymal vimentin but do not express markers associated with myoepithelial cells or fibroblasts. Treatment with estradiol does not affect either the proliferation rate or the induction of gene expression in MCF-12A cells. Parental MCF-12A cells form acini, solid spheres and elongated branching ducts when grown in rat-tail collagen type I matrix, the geometries and distribution of which are altered following the removal of fibroblast-like cells. MCF-12A cells are a heterogeneous pseudo-epithelial cell line capable of forming a variety of multicellular structures in 3D culture. We found no indication that the cells display estrogen-responsive characteristics, thus refuting previous studies which reported estrogen responsiveness. We report that MCF-12A cells are not suited for use in studies in which differential behaviors of "normal" and "cancerous" estrogen-responsive cells are to be compared.


[The topographic mapping of evoked bioelectrical activity and other methods for the functional neural visualization of the brain].
The paper is concerned with comparative characterization of brain functional imaging used in medicine today: bioelectrical activity mapping, assessment of spatial changes in cerebral blood flow by radionuclide and hydrogen clearance, NMR imaging, PET-scan, magnetoencephalography and thermoencephaloscopy. Original researches in neurosurgery clinic with the use of the mapping of evoked potentials enable the conclusion to be drawn about advisability of applying these methods in treatment and diagnostic practice. Besides, the mechanisms of compensation for higher mental functions in organic brain lesions are discussed.


Biliary Stenting in Patients With Pancreatic Cancer: Results From a Population-Based Cohort Study.
We aimed to describe management of biliary obstruction (BO) in the context of pancreatic cancer within a population-based cohort. We examined management of BO in 1863 patients diagnosed as having pancreatic cancer in 2010/2011. We used descriptive statistics and logistic regression to describe patterns of biliary stent usage, complications and duration of patency, associations between preoperative stenting and surgical outcomes, and between patient factors and management of jaundice. Almost half of the people in the cohort (n = 909) were jaundiced within 12 months of diagnosis. Two-thirds of these had at least 1 stent inserted. Preoperative stenting, mostly with plastic stents, occurred for 72% of patients who experienced jaundice prior to an attempted resection but was not associated with surgical outcomes. Seventy percent of the jaundiced patients who did not have an attempted resection were stented. Metal stents were less frequently replaced within 30 days than plastic (9% vs 42%). Living in a rural area was associated with reduced likelihood of having jaundice managed. Plastic stents were still used frequently, despite guidelines recommending metal in most contexts. Patients living in rural areas were less likely to have BO managed. This work highlights the need to monitor current practice.


Bioaugmentation with Endophytic Bacterium E6S Homologous to Achromobacter piechaudii Enhances Metal Rhizoaccumulation in Host Sedum plumbizincicola.
Application of hyperaccumulator-endophyte symbiotic systems is a potential approach to improve phytoremediation efficiency, since some beneficial endophytic bacteria are able to detoxify heavy metals, alter metal solubility in soil, and facilitate plant growth. The objective of this study was to isolate multi-metal resistant and plant beneficial endophytic bacteria and to evaluate their role in enhancing plant growth and metal accumulation/translocation. The metal resistant endophytic bacterial strain E6S was isolated from stems of the Zn/Cd hyperaccumulator plant Sedum plumbizincicola growing in metalliferous mine soils using Dworkin and Foster salts minimal agar medium with 1-aminocyclopropane-1-carboxylate (ACC) as the sole nitrogen source, and identified as homologous to Achromobacter piechaudii based on morphological and biochemical characteristics, partial 16S rDNA sequence and phylogenetic analysis. Strain E6S showed high level of resistance to various metals (Cd, Zn, and Pb). Besides utilizing ACC, strain E6S exhibited plant beneficial traits, such as solubilization of phosphate and production of indole-3-acetic acid. Inoculation with E6S significantly increased the bioavailability of Cd, Zn, and Pb in soil. In addition, bacterial cells bound considerable amounts of metal ions in the following order: Zn > Cd >Pb. Inoculation of E6S significantly stimulated plant biomass, uptake and bioaccumulation of Cd, Zn, and Pb. However, E6S greatly reduced the root to shoot translocation of Cd and Zn, indicating that bacterial inoculation assisted the host plant to uptake and store heavy metals in its root system. Inoculation with the endophytic bacterium E6S homologous to A. piechaudii can improve phytostabilization of metalliferous soils due to its effective ability to enhance in situ metal rhizoaccumulation in plants.


Endocrine and neuroendocrine regulation of social status in cichlid fishes.
Position in a dominance hierarchy profoundly impacts group members' survival, health, and reproductive success. Thus, understanding the mechanisms that regulate or are associated with an individuals' social position is important. Across taxa, various endocrine and neuroendocrine signaling systems are implicated in the control of social rank. Cichlid fishes, with their often-limited resources of food, shelter, and mates that leads to competition, have provided important insights on the proximate and ultimate mechanisms related to establishment and maintenance of dominance hierarchies. Here we review the existing information on the relationships between endocrine (e.g., circulating hormones, gonadal and other tissue measures) and neuroendocrine (e.g., central neuropeptides, biogenic amines, steroids) systems and dominant and subordinate social rank in male cichlids. Much of the current literature is focused on only a few representative cichlids, particularly the African Astatotilapia burtoni, and several other African and Neotropical species. Many hormonal regulators show distinct differences at multiple biological levels between dominant and subordinate males, but generalizations are complicated by variations in experimental paradigms, methodological approaches, and in the reproductive and parental care strategies of the study species. Future studies that capitalize on the diversity of hierarchical structures among cichlids should provide insights towards better understanding the endocrine and neuroendocrine mechanisms contributing to social rank. Further, examination of this topic in cichlids will help reveal the selective pressures driving the evolution of endocrine-related phenotypic traits that may facilitate an individual's ability to acquire and maintain a specific social rank to improve survival and reproductive success.


Policy choice or economic fundamentals: what drives the public-private health expenditure balance in Canada?
The public share of Canadian total health expenditure declined between 1975 and 2005 but categories such as physicians stayed constant while drugs, other institutions, and capital spending saw increases. Regressions find the key determinants of the public share to be: the relative price of health care, per capita income, federal transfers, proportion of population over age 65, provincial dummy variables, political parties, the onset of the Canada Health Act and the Canada Health and Social Transfer (CHST), and a time trend invoking technological change. Increasing income inequality is not a factor eroding the public share. Moreover, provinces governed by center-left parties are associated with lower public shares in the physician and other health professional categories. The significance of variables like the Canada Health and Social Transfer, the Canada Health Act and provincial differences suggest the increase in the private share of health spending since 1975 is partly the result of policy choices. As the public share has only declined from approximately 76% to 70% over 30 years, it appears the shift towards private care is one of marginal increments. Canadians remain in conflict over their health care system as they appear willing to tolerate only marginal accretions in the overall private share of health spending but accept larger changes across health expenditure categories and provincial systems.


Positional repeatability and variation in internal and external markers during volumetric-modulated arc therapy under end-exhalation breath-hold conditions for pancreatic cancer patients.
The purpose of this study was to assess the positional repeatability of internal and external markers among multiple breath-hold (BH) sessions and evaluate the positional variation of these markers within BH sessions for volumetric-modulated arc therapy (VMAT) for pancreatic cancer patients. A total of 13 consecutive pancreatic cancer patients with an internal marker were enrolled. Single full-arc coplanar VMAT was delivered under end-exhalation BH conditions while monitoring the internal marker with kilovoltage (kV) X-ray fluoroscopy. Positional repeatability of the internal and external markers was determined by the difference between the reference and zero position in all BH sessions, and positional variation was defined by the displacement from the reference position in each BH session during megavolt beam delivery. The overall positional repeatability was 0.6 ± 1.5 mm in the X-axis for the centroid of the internal marker (CoIM), -0.1 ± 2.2 mm in the Y-axis for the CoIM, and 0.8 ± 2.2 mm for the external marker. The frequency of an internal marker position appearing > 2 mm from the reference position in the Y-axis, despite the external marker position being ≤2 mm from the reference position, ranged from 0.0 to 39.9% for each patient. Meanwhile, the proportion of sessions with positional variation ≤2 mm was 93.2 and 98.7% for the CoIM and external marker, respectively. External marker motion can be used as a surrogate for pancreatic tumor motion during BH-VMAT delivery; however, margins of ~5 mm were required to ensure positional repeatability.


Carotid artery revascularization.
Stroke is the third leading cause of death in the United States, and up to one third of patients have a stroke secondary to carotid occlusive disease. Surgical management has firmly established itself as an important modality in treating this disease. Several prospective randomized trials have defined the patients that would have the most benefit from carotid endarterectomy (CEA). These patient populations include asymptomatic patients with a >or= 60% stenosis and symptomatic patients with a >or= 50% stenosis. The timing of CEA after stroke remains controversial, but recent studies advocate early CEA in a select group of patients. During the CEA, the method of closing of the arteriotomy has an overall effect on the safety of the procedure as well as long-term outcome. As compared with primary repair of the arteriotomy, patch closure has been shown to decrease the frequency of restenosis. In addition, carotid eversion endarterectomy (CEE) is an alternative method to remove the plaque that has a similar efficacy to standard CEA. The role of carotid angioplasty and stenting (CAS) continues to evolve and offers the patient a less invasive method of treating the carotid plaque.


Free Cholesterol Accelerates Aβ Self-Assembly on Membranes at Physiological Concentration.
The effects of membranes on the early-stage aggregation of amyloid β (Aβ) have come to light as potential mechanisms by which neurotoxic species are formed in Alzheimer's disease. We have shown that direct Aβ-membrane interactions dramatically enhance the Aβ aggregation, allowing for oligomer assembly at physiologically low concentrations of the monomer. Membrane composition is also a crucial factor in this process. Our results showed that apart from phospholipids composition, cholesterol in membranes significantly enhances the aggregation kinetics. It has been reported that free cholesterol is present in plaques. Here we report that free cholesterol, along with its presence inside the membrane, further accelerate the aggregation process by producing aggregates more rapidly and of significantly larger sizes. These aggregates, which are formed on the lipid bilayer, are able to dissociate from the surface and accumulate in the bulk solution; the presence of free cholesterol accelerates this dissociation as well. All-atom molecular dynamics simulations show that cholesterol binds Aβ monomers and significantly changes the conformational sampling of Aβ monomer; more than doubling the fraction of low-energy conformations compared to those in the absence of cholesterol, which can contribute to the aggregation process. The results indicate that Aβ-lipid interaction is an important factor in the disease prone amyloid assembly process.


[Theory and practice in developing a system of artificial intelligence for recognizing histologic specimens].
A system is described for an automatic analysis and recognition of a fresh and reference histologic material. The system includes a microscope, TV camera and personal computer. The system proved applicable in identification of two tumor types: astrocytomas and oligodendrogliomas. The system can be used in pathology, histology, embryology, cytology, histochemistry, clinical laboratory microscopy and in borderline fields.


Analysis of the structural genes encoding M-factor in the fission yeast Schizosaccharomyces pombe: identification of a third gene, mfm3.
We previously identified two genes, mfm1 and mfm2, with the potential to encode the M-factor mating pheromone of the fission yeast Schizosaccharomyces pombe (J. Davey, EMBO J. 11:951-960, 1992), but further analysis revealed that a mutant strain lacking both genes still produced active M-factor. Here we describe the isolation and characterization of a third M-factor gene, mfm3. A mutant lacking all three genes fails to produce M-factor, indicating that all functional M-factor genes now have been identified. The triple mutant exhibits an absolute mating defect in M cells, a defect that is not rescued by addition of exogenous M-factor. A mutational analysis reveals that all three mfm genes contribute to the production of M-factor. Their transcription is limited to M cells and requires the mat1-Mc and ste11 gene products. Each gene is induced when the cells are starved of nitrogen and further induced by a pheromone signal. Additionally, the signal transduction machinery associated with the pheromone response is required for transcription of the mfm genes in both stimulated and unstimulated cells.


[The structure of the germinal centers of the thymus].
A morphological investigation of the thymus was performed in 82 patients dead from rheumatism. The germinal centers of the thymus localized in the cortical layer and medullary substance were found in 42% of observations. These centers were not homogeneous in their structure. Some of them represent germinal centers of lymphoid nodules, others have epithelioreticulocytes. The latters having a substantial difference from the centers of the lymphoid nodules should be referred to as thymic islands. Necrosis of their cells appear in these islands, its spreading being responsible for transformation of the islands in cysts. The appearance of islands and the following necrosis are thought to be a mechanism of elimination of the thymus cells.


Structural determinants of macrocyclization in substrate-controlled lanthipeptide biosynthetic pathways.
Lanthipeptides are characterized by thioether crosslinks formed by post-translational modifications. The cyclization process that favors a single ring pattern over many other possible ring patterns has been the topic of much speculation. Recent studies suggest that for some systems the cyclization pattern and stereochemistry is determined not by the enzyme, but by the sequence of the precursor peptide. However, the factors that govern the outcome of the cyclization process are not understood. This study presents the three-dimensional structures of seven lanthipeptides determined by nuclear magnetic resonance spectroscopy, including five prochlorosins and the two peptides that make up cytolysin, a virulence factor produced by Enterococcus faecalis that is directly linked to human disease. These peptides were chosen because their substrate sequence determines either the ring pattern (prochlorosins) or the stereochemistry of cyclization (cytolysins). We present the structures of prochlorosins 1.1, 2.1, 2.8, 2.10 and 2.11, the first three-dimensional structures of prochlorosins. Our findings provide insights into the molecular determinants of cyclization as well as why some prochlorosins may be better starting points for library generation than others. The structures of the large and small subunits of the enterococcal cytolysin show that these peptides have long helical stretches, a rare observation for lanthipeptides characterized to date. These helices may explain their pore forming activity and suggest that the small subunit may recognize a molecular target followed by recruitment of the large subunit to span the membrane.


Surgical treatment of hepatocellular carcinoma with bile duct tumor thrombi.
To study the surgical treatment effect and outcome of hepatocellular carcinoma (HCC) with bile duct tumor thrombi (BDTT). Fifty-three consecutive HCC patients with BDTT admitted in our department from July 1984 to December 2002 were reviewed retrospectively. The clinical data, diagnostic methods, surgical procedures and outcome of these patients were collected and analyzed. One patient rejected surgical treatment, 6 cases underwent percutaneous transhepatic cholangial drainage (PTCD) for unresectable primary disease, and the other 46 cases underwent surgical operation. The postoperative mortality was 17.6%, and the morbidity was 32.6%. Serum total bilirubin levels of these patients with obstructive jaundice decreased gradually after surgery. The survival time of six cases who underwent PTCD ranged from 2 to 7 mo (median survival of 3.7 mo). The survival time of the patients who received surgery was as follows: 2 mo for one patient who underwent laparotomy, 5-46 mo (median survival of 23.5 mo, which was the longest survival in comparison with patients who underwent other procedures, P = 0.0024) for 17 cases who underwent hepatectomy, 5-17 mo (median survival of 10.0 mo) for 5 cases who underwent HACE, 3-9 mo (median survival of 6.1 mo) for 11 cases who underwent simple thrombectomy and biliary drainage, and 3-8 mo (median survival of 4.3 mo) for four cases who underwent simple biliary drainage. Jaundice caused by BDTT in HCC patients is not a contraindication for surgery. Only curative resection can result in long-term survival. Early diagnosis and surgical treatment are the key points to prolong the survival of patients.


Cardiovascular safety trials of incretin-based drugs: What do they mean?
Incretin-based dipeptidyl peptidase-4 inhibitors and glucagon-like peptide-1 receptor agonists are newer choices of antidiabetic medications that are now most widely used worldwide. Preclinical study results suggest that the two drugs potentially exert benefits to prevent onsets and/or progressions of diabetes-related complications, such as myocardial infarctions and strokes. Outcomes of five clinical trials to evaluate the cardiovascular (CV) safety of dipeptidyl peptidase-4 inhibitors and glucagon-like peptide-1 receptor agonist have been recently reported. The heart failure findings of the Saxagliptin Assessment of Vascular Outcomes Recorded in Patients with Diabetes Mellitus-Thrombolysis in Myocardial Infarction 53 (SAVOR-TIMI53) are unexpected and very concerning; results of the Examination of Cardiovascular Outcomes with Alogliptin vs Standard of Care (EXAMINE), the Trial Evaluating Cardiovascular Outcomes with Sitagliptin (TECOS) and the Evaluation of Lixisenatide in Acute Coronary Syndrome (ELIXA) encourage neutral CV safety profiles of incretin-based drugs in individuals with type 2 diabetes and established CV diseases or multiple CV risks. Furthermore, the Liraglutide Effect and Action in Diabetes: Evaluation of Cardiovascular Outcome Results (LEADER) results show the benefits of liraglutide in preventing CV events in a similar study population. Despite the many preclinical studies showing the beneficial effects of incretin-related drugs, most CV safety trials of incretin-based drugs, except for LEADER, did not show benefits for CV events. It is important to recognize that CV safety trials were carried out to meet the US Food and Drug Administration guidance to assess CV safety of all new antidiabetic drugs; they were not designed to assess their benefits for CV events. Therefore, the long-term potential benefit, as well as even the safety, of incretin-based drugs for certain CV outcomes has not been definitively established, and requires evaluation in more specific and more relevant trials. If the need for CV safety trials would be determined based on an individual drug's safety data during its earlier development as well as its mechanism of action, resources could be saved for carrying out such clinical trials.


Pseudocercospora and allied genera associated with leaf spots of banana (Musa spp.).
The Sigatoka leaf spot complex on Musa spp. includes three major pathogens: Pseudocercospora, namely P. musae (Sigatoka leaf spot or yellow Sigatoka), P. eumusae (eumusae leaf spot disease), and P. fijiensis (black leaf streak disease or black Sigatoka). However, more than 30 species of Mycosphaerellaceae have been associated with Sigatoka leaf spots of banana, and previous reports of P. musae and P. eumusae need to be re-evaluated in light of recently described species. The aim of the present study was thus to investigate a global set of 228 isolates of P. musae, P. eumusae and close relatives on banana using multigene DNA sequence data [internal transcribed spacer regions with intervening 5.8S nrRNA gene (ITS), RNA polymerase II second largest subunit gene (rpb2), translation elongation factor 1-alpha gene (tef1), beta-tubulin gene (tub2), and the actin gene (act)] to confirm if these isolates represent P. musae, or a closely allied species. Based on these data one new species is described, namely P. pseudomusae, which is associated with leaf spot symptoms resembling those of P. musae on Musa in Indonesia. Furthermore, P. eumusae, P. musae and P. fijiensis are shown to be well defined taxa, with some isolates also representing P. longispora. Other genera encountered in the dataset are species of Zasmidium (Taiwan leaf speckle), Metulocladosporiella (Cladosporium leaf speckle) and Scolecobasidium leaf speckle. Citation: Crous P, Carlier J, Roussel V, Groenewald JZ (2020). Pseudocercospora and allied genera associated with leaf spots of banana (Musa spp.). Fungal Systematics and Evolution 7: 1-19. doi: 10.3114/fuse.2021.07.01.


Nitrogen fixation catalyzed by ferrocene-substituted dinitrogen-bridged dimolybdenum-dinitrogen complexes: unique behavior of ferrocene moiety as redox active site.
A series of dinitrogen-bridged dimolybdenum-dinitrogen complexes bearing metallocene-substituted PNP-pincer ligands is synthesized by the reduction of the corresponding monomeric molybdenum-trichloride complexes under 1 atm of molecular dinitrogen. Introduction of ferrocene as a redox-active moiety to the pyridine ring of the PNP-pincer ligand increases the catalytic activity for the formation of ammonia from molecular dinitrogen, up to 45 equiv. of ammonia being formed based on the catalyst (22 equiv. of ammonia based on each molybdenum atom of the catalyst). The time profile for the catalytic reaction reveals that the presence of the ferrocene unit in the catalyst increases the rate of ammonia formation. Electrochemical measurement and theoretical studies indicate that an interaction between the Fe atom of the ferrocene moiety and the Mo atom in the catalyst may play an important role to achieve a high catalytic activity.


Gallstone Ileus in Octogenarians: Is Cholecystectomy Really Needed?
Gallstone ileus is an uncommon complication of cholelithiasis and occurs when a gallstone migrates through a cholecystoenteric fistula and impacts within the gastrointestinal tract. Surgical intervention remains the treatment of choice, which consists of a full-thickness incision of the visceral wall and removal of the impacted gallstone. In this paper we present the treatment approach of 6 cases of gallstone ileus in octogenarians. In our cohort, intestinal obstruction was resolved through an enterotomy or gastrotomy and lithotomy/stone extraction in every patient. No cholecystectomies were undertaken. Despite the fact that gallstone ileus is diagnosed in small percent of patients suffering from gallstone disease, it accounts for a large proportion of intestine obstruction in patients older than 65 years old. Since accurate diagnosis and timely intervention are vital, providers should be familiar with the diagnostic approach and the treatment of this clinical entity.


Health Belief Model efficacy in explaining and predicting intention or uptake pertussis vaccination during pregnancy.
Pertussis is a highly contagious respiratory disease and vaccination of pregnant women seems to be the most effective strategy to prevent pertussis in infants. The aim of this study is to assess the acceptance by women of pertussis vaccination during pregnancy based on Health Belief Model (HBM) constructs. A multicentre observational study was carried out with a convenience sample of 300 respondents. Most women were worried to contract or to transmit pertussis during the first months of the infant's life and perceived pertussis contracted in the first months of life as very serious. Parity appears to be a factor predicting this health behaviour, as nulliparous women tend to get more vaccinated or have a higher intention to get vaccinated (ORa 2.8 CI 95% 1.5-5.2 p<0.01). HBM is an effective tool for identifying facilitators and barriers to health behaviours. Strategies to promote vaccination during pregnancy are needed, including educational interventions and communication campaigns.


Does visual flicker phase at gamma frequency modulate neural signal propagation and stimulus selection?
Oscillatory synchronization of neuronal populations has been proposed to play a role in perceptual integration and attentional processing. However, some conflicting evidence has been found with respect to its causal relevance for sensory processing, particularly when using flickering visual stimuli with the aim of driving oscillations. We tested psychophysically whether the relative phase of gamma frequency flicker (60 Hz) between stimuli modulates well-known facilitatory lateral interactions between collinear Gabor patches (Experiment 1) or crowding of a peripheral target by irrelevant distractors (Experiment 2). Experiment 1 assessed the impact of suprathreshold Gabor flankers on detection of a near-threshold central Gabor target ("Lateral interactions paradigm"). The flanking stimuli could flicker either in phase or in anti-phase with each other. The typical facilitation of target detection was found with collinear flankers, but this was unaffected by flicker phase. Experiment 2 employed a "crowding" paradigm, where orientation discrimination of a peripheral target Gabor patch is disrupted when surrounded by irrelevant distractors. We found the usual crowding effect, which declined with spatial separation, but this was unaffected by relative flicker phase between target and distractors at all separations. These results imply that externally driven manipulations of gamma frequency phase cannot modulate perceptual integration in vision.


Mechano-optical wavelength tuning in a photonic crystal microcavity with sub-1 V drive voltage.
A micro-bimorph cantilever with self-aligned nanotips is monolithically integrated with a photonic crystal based device using optical and deep UV lithography techniques. Upon electrostatic actuation, the dielectric nanotips perturb the optical field, providing electromechano-optical modulation of light. Static tuning of the optical transmission spectra by more than 600 pm is measured with a sub-1 V drive voltage, resulting in a modulation as high as 21 dB. The observed strong electromechano-optical effect may find application in power efficient devices for optical communication networks, such as wavelength routing elements.


Urodynamic alterations after pelvic floor exercises for treatment of stress urinary incontinence in women.
The following study improves the effectiveness of pelvic floor exercise in women with stress urinary incontinence. A group of 27 women with stress urinary incontinence performed specific pelvic floor exercises twice a week for 45 minutes under a therapist's supervision for a period of 12 weeks. This group had urodynamic and urethral pressure profile studies and filled in a daily diary. The patients self-evaluated their symptoms. After the therapy, the urodynamic and urethreal pressure profile studies were repeated and the results were: 66.7% patients were self-evaluated as cured; 14.8% improved and 18.5% unchanged. The urodynamic results showed that 48.2 % of the patients did not have urinary loss, however, 51,8% of the patients that had a loss showed an increase in vesicle volume, and only 7.3% remained unchanged. The results show that pelvic floor exercises are an effective and low cost treatment for stress urinary incontinence rehabilitation.


Community-acquired pneumonia: epidemiologic and clinical considerations.
An incidence of between 2 and 44 per 1000 population has been reported for community-acquired pneumonia. Epidemiologic studies describe a wide range of causative organisms, including Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae, Legionella spp., Moraxella catarrhalis, Chlamydia pneumoniae and viruses such as influenza A and B. However, the frequency with which they are reported varies widely. On analysis of these studies, the variation can be explained by a number of factors. The results depend on the definition of pneumonia and the criteria for assigning a causative role to any particular organism. Older studies have not included diagnostic methods for newly described organisms such as C. pneumoniae and Legionella spp. The improved diagnostic methods for these organisms and for Mycoplasma pneumoniae are reflected in more recent studies. Further variation depends on the population studied. As many patients with mild pneumonia are successfully treated in the community, those studies that are hospital-based include patients with more severe pneumonia often in the elderly or in patients with underlying diseases such as chronic obstructive pulmonary disease. The prior use of antibiotics not only contributes to the high percentage of cases for which no etiologic agent is found, but also ensures that treatment failures are selected for hospitalization. This further changes the result, depending on the antibiotic agents used most commonly in the community. The inclusion of nursing home patients or groups where alcoholism is more common will also favor particular organisms. Finally, the timing of the study may be such that an epidemic is included. This has relevance mostly for Mycoplasma pneumoniae, C. pneumoniae, Legionella spp. and influenza. In the assessment of the patient with community-acquired pneumonia, any one of the above organisms can be considered to be responsible. As initial treatment is empirical, other information can be used to ensure that an antibiotic with an adequate spectrum is chosen. Factors of importance are age, underlying illness, severity of disease and any locally recognized epidemics or endemic organisms. Differences in clinical presentation are not sufficiently distinct to allow for accurate prediction of the causative agent. Similarly, chest radiograph changes are not sufficiently specific to discriminate reliably between diverse organisms such as S. pneumoniae, Mycoplasma pneumoniae and Legionella spp. Current recommendations for choice of an empirical antibiotic agent are therefore based, not on the assumption of a single etiologic agent indicated by clinical presentation or radiographic appearances, but on age of the patient, severity of illness, the presence of underlying conditions and the range of possible organisms in that patient group.


Mood Stabilizers and the Influence on Global Leukocyte DNA Methylation in Bipolar Disorder.
Little is known about the relationship between treatments for bipolar disorder (BD), their therapeutic responses and the DNA methylation status. We investigated whether global DNA methylation levels differ between healthy controls and bipolar patients under different treatments. Global DNA methylation was measured in leukocyte DNA from bipolar patients under lithium monotherapy (n = 29) or combination therapy (n = 32) and from healthy controls (n = 26). Lithium response was assessed using the Alda scale. Lithium in monotherapy was associated with hypomethylation (F = 4.63, p = 0.036). Lithium + valproate showed a hypermethylated pattern compared to lithium alone (F = 7.27, p = 0.011). Lithium response was not associated with DNA methylation levels. These data suggest that the choice of treatment in BD may lead to different levels of global DNA methylation. However, further research is needed to understand its clinical significance.


Comparison of the structures of three carboxypeptidase A-phosphonate complexes determined by X-ray crystallography.
The structures of the complexes of carboxypeptidase A (CPA) with two tight-binding phosphonate inhibitors have been determined by X-ray crystallography. The inhibitors, Cbz-Phe-ValP-(O)-Phe[ZFVP(O)F] and Cbz-Ala-GlyP-(O)-Phe[ZAGP(O)F], bind noncovalently to CPA with dissociation constants (Ki's) of 11 fM and 710 pM, respectively. The CPA-ZFVP(O)F complex crystallizes in the space group P2(1)2(1)2(1) with unit cell parameters a = 65.3 A, b = 63.4 A, and c = 76.0 A, and the CPA-ZAGP(O)F complex crystallizes in the space group P2(1)2(1)2(1) with unit cell parameters a = 63.4 A, b = 65.9 A, and c = 74.4 A. Both structures were determined by molecular replacement to a resolution of 2.0 A. The final crystallographic residuals are 0.189 for the CPA-ZFVP(O)F complex and 0.191 for the CPA-ZAGP(O)F complex. The CPA-ZFVP(O)F complex exhibits the lowest Ki yet determined for an enzyme-inhibitor interaction. Comparison of the CPA-ZFVP(O)F structure with that of the CPA-ZAAP(O)F complex [Kim, H., & Lipscomb, W.N. (1990) Biochemistry 29, 5546-5555] indicates the likely important contributions of hydrophobic and weakly polar enzyme-inhibitor interactions to the exceptional stability of the CPA-ZFVP(O)F complex. Among these interactions is a network of four aromatic rings of CPA and ZFVP(O)F in a configuration that allows stabilizing aromatic-aromatic edge-to-face interactions from one ring to the next. A comparison of the structures of the CPA-ZFVP(O)F, CPA-ZAAP(O)F and CPA-ZAGP(O)F complexes shows that all three phosphonates assume a similar binding mode in the active-site binding groove of CPA. For ZAGP(O)F, the glycyl P1 residue does not lead to an anomalous or a partially disordered binding mode as seen in some previous complexes of CPA involving dipeptide analogue inhibitors with glycyl P1 residues. The additional enzyme-inhibitor interactions for these tripeptide phosphonates secure a binding mode in which a Pi portion of the inhibitor is clearly bound by the corresponding Si binding subsite. These three phosphonates have been implicated as transition-state analogues of the CPA-catalyzed reaction. The phosphinyl groups of these phosphonates coordinate to the active-site zinc in a manner that has been proposed as a characteristic feature of the general-base (Zn-hydroxyl or Zn-water) mechanism for the CPA-catalyzed reaction. Further mechanistic proposals are made for Arg-127, whose probable role in binding substrates is apparent in these CPA-phosphonate complexes.


Discovery of the novel autophagy inhibitor aumitin that targets mitochondrial complex I.
Macroautophagy is a conserved eukaryotic process for degradation of cellular components in response to lack of nutrients. It is involved in the development of diseases, notably cancer and neurological disorders including Parkinson's disease. Small molecule autophagy modulators have proven to be valuable tools to dissect and interrogate this crucial metabolic pathway and are in high demand. Phenotypic screening for autophagy inhibitors led to the discovery of the novel autophagy inhibitor aumitin. Target identification and confirmation revealed that aumitin inhibits mitochondrial respiration by targeting complex I. We show that inhibition of autophagy by impairment of mitochondrial respiration is general for several mitochondrial inhibitors that target different mitochondrial complexes. Our findings highlight the importance of mitochondrial respiration for autophagy regulation.


iPhos: a toolkit to streamline the alkaline phosphatase-assisted comprehensive LC-MS phosphoproteome investigation.
Comprehensive characterization of the phosphoproteome in living cells is critical in signal transduction research. But the low abundance of phosphopeptides among the total proteome in cells remains an obstacle in mass spectrometry-based proteomic analysis. To provide a solution, an alternative analytic strategy to confidently identify phosphorylated peptides by using the alkaline phosphatase (AP) treatment combined with high-resolution mass spectrometry was provided. While the process is applicable, the key integration along the pipeline was mostly done by tedious manual work. We developed a software toolkit, iPhos, to facilitate and streamline the work-flow of AP-assisted phosphoproteome characterization. The iPhos tookit includes one assister and three modules. The iPhos Peak Extraction Assister automates the batch mode peak extraction for multiple liquid chromatography mass spectrometry (LC-MS) runs. iPhos Module-1 can process the peak lists extracted from the LC-MS analyses derived from the original and dephosphorylated samples to mine out potential phosphorylated peptide signals based on mass shift caused by the loss of some multiples of phosphate groups. And iPhos Module-2 provides customized inclusion lists with peak retention time windows for subsequent targeted LC-MS/MS experiments. Finally, iPhos Module-3 facilitates to link the peptide identifications from protein search engines to the quantification results from pattern-based label-free quantification tools. We further demonstrated the utility of the iPhos toolkit on the data of human metastatic lung cancer cells (CL1-5). In the comparison study of the control group of CL1-5 cell lysates and the treatment group of dasatinib-treated CL1-5 cell lysates, we demonstrated the applicability of the iPhos toolkit and reported the experimental results based on the iPhos-facilitated phosphoproteome investigation. And further, we also compared the strategy with pure DDA-based LC-MS/MS phosphoproteome investigation. The results of iPhos-facilitated targeted LC-MS/MS analysis convey more thorough and confident phosphopeptide identification than the results of pure DDA-based analysis.


Temperature and protein kinase C modulate myofilament Ca2+ sensitivity in pressurized rat cerebral arteries.
The effects of pharmacological activation and inhibition of protein kinase C (PKC) and temperature on the relationship between cytoplasmic Ca2+ and lumen diameter were studied in pressurized (50 mmHg) rat posterior cerebral arteries permeabilized with alpha-toxin. Increasing Ca2+ concentrations (30 nM-10 microM, 22 degrees C) induced stable, concentration-dependent constrictions with a half-maximal effective concentration (EC50) of 112 nM. The maximal constriction was 80% of baseline diameter and 157% of that during depolarization of nonpermeabilized vessels with 124 mM KCl. Elevation of temperature to 37 degrees C increased the EC50 to 246 nM and enhanced the steepness of concentration-response curves. Exposure of permeabilized arteries to indolactam V, an activator of PKC, resulted in a significant myofilament Ca2+ sensitization (e.g., EC50 at 5 microM = 126 nM) without changing efficacy. The effects of calphostin C, a PKC inhibitor, on Ca2+ sensitivity were minimal; however, the amplitude of Ca2+-induced constrictions in both control and indolactam-treated arteries was suppressed in a concentration-dependent manner. Thus 1) temperature is an important variable in studies of arterial Ca2+ sensitivity, and 2) changes in PKC activity can significantly alter both myofilament sensitivity to and constrictor efficacy of cytosolic Ca2+.


Napping at home and alertness on the job in rotating shift workers.
Conditions surrounding self-selected napping and its association with waking function were examined in a quasi-experimental study at two worksites with workers on 8- and 12-hour rotating shifts. Nap frequency, sleep quantity, quality and timing were determined from daily questionnaires, and performance/alertness during the workshift was assessed with on-site computerized testing. Results indicated that most napping occurred prior to the first night shift of the week. Napping prior to the first shift supplemented an apparently adequate quantity of main sleep, whereas napping during the workweek compensated for a reduced quantity of main sleep. Sleep quality was rated higher on no-nap days than on nap days and higher prior to the first shift than during the workweek. These results generally support a compensatory view of napping. Worksite performance/alertness testing, however, indicated diminished alertness during the shift on nap days compared to no-nap days, which was not consistent with a compensatory view of napping. Decreased alertness on nap days may have been associated with poorer sleep quality or with differences in circadian rhythm adaptation on those days.


Combined antileukemic activity of pIXY 321 and Ara-C against human acute myeloid leukemia cells.
Prolonged administration of conventional (100 mg/m2/day) or low dose Ara-C (20 mg/m2/day) has been associated with significant clinical antileukemic effects in AML and myelodysplastic syndromes. These doses and schedules of Ara-C yield plasma Ara-C concentrations in the range of 10 to 100 nM. Utilizing concentrations and a schedule of Ara-C treatment, representative of Ara-C exposures in these clinical situations, we performed in vitro studies to examine the effects of co-treatment with pIXY 321 on Ara-C induced apoptosis and Ara-C-mediated colony growth inhibition of human myeloid leukemia HL-60 cells. Significantly greater internucleosomal DNA fragmentation, higher percentage of morphologically recognizable apoptotic cells and increased colony growth inhibition were observed following treatment with 100 versus 10 nM Ara-C for 5 days. Simultaneous exposure to 10 ng/ml pIXY 321 resulted in significantly increased colony growth inhibition as well as DNA fragmentation and apoptosis due to 10 nM but not 100 nM Ara-C. These concentrations of Ara-C inhibited c-myc and did not induce c-jun mRNA expression. These effects of Ara-C on c-myc and c-jun expressions were not influenced by co-treatment with pIXY 321. Neither treatment with pIXY 321 or Ara-C alone, nor co-treatment with pIXY 321 and Ara-C, significantly altered the intracellular p26BCL-2 levels in HL-60 cells. These results indicate that co-treatment with pIXY 321 significantly increases low dose Ara-C-induced apoptosis and thereby its antileukemic activity.


Anisotropic exchange interaction and field-induced SMM behaviour in a mixed valence {CoCo} complex.
We are reporting the synthesis and structural characterization of a new hexanuclear Co(ii)/Co(iii) complex starting from a versatile pivalate cobalt precursor and the racemic mixture of a chelating Schiff base type ligand. The main [CoII4CoIII2(μ3-OH)2(μ-OR)2(μ-OR')2(μ-OR'')2]6+ core is unprecedented and exhibits an inversion center that affords only two unique Co(ii) sites. We performed DC and AC magnetic measurements and analysed them in terms of the anisotropic exchange of ground Kramers doublets at each Co(ii) site due to their unquenched angular orbital contribution to the magnetic moment. Quantum computations support the experimental data treatment. The interplay of dominant antiferromagnetic exchange, inversion symmetry and a non-collinear main quantization axis affords an exchange energy spectrum with mostly non-magnetic states. Nevertheless, field induced SMM behaviour is observed at 1500 Oe and below 3 K which might be explained by the relaxation of the first excited magnetic state (which is populated enough) through the next closest excited state. The Orbach and/or Raman mechanism could be operative from the experimental and quantum computed results.


The function of microRNA-211 expression in post-fracture bone cell apoptosis involving the transforming growth factor-β/ phosphoinositide 3-kinase signaling pathway.
The underlying mechanism of micro (mi)RNA-211 in bone cell apoptosis after fracture remains unclear. This study aimed to determine the effect and function of miRNA-211 in bone cell apoptosis in fracture patients. Serum samples were collected from patients with fractures and healthy controls. Serum miR-211 expression was detected by quantitative PCR. MC3T3-E1 cells were transfected with a transforming growth factor (TGF)-β inhibitor and phosphoinositide 3-kinase (PI3K) inhibitor. The viability of MC3T3-E1 cells was detected by the MTT assay, and apoptosis was detected by flow cytometry. Caspase-3/9 activity and the protein expression of TGF-β, PI3K, and p-Akt were detected by western blot and immunoprecipitation. In the fracture group, miRNA-211 expression was significantly up-regulated compared with controls. We used miRNA-211 mimics to up-regulate miRNA-211 expression, and observed inhibited cell viability and induced apoptosis and lactate dehydrogenase (LDH) activity. miRNA-211 up-regulation also suppressed the expression of TGF-β, PI3K, and p-Akt proteins. Conversely, miRNA-211 down-regulation increased cell viability and reduced apoptosis and LDH activity, as well as inducing the expression of TGF-β, PI3K, and p-Akt. Inhibiting TGF-β decreased the effect of anti-miRNA-211 on osteocyte apoptosis. Our data indicate that miRNA-211 functions via the TGF-β/PI3K/Akt signaling pathway in patients with fractures.


An educator-administered measure of language development in young children.
An increasing number of infants and toddlers in many countries are enrolled in early childhood education (ECE) programs, and educators thus play a central role in stimulating language development in these young children. A valid, brief educator-completed measure of language development in young children has important uses both for the identification and monitoring of language development and for the guidance and evaluation of intentional instruction and targeted interventions for children who need it. We present such a measure here for Danish, the CDI: Educator (CDI-Edu) version, which is based on well-developed and validated parent report measures, adapted for the early childhood education setting. It requires approximately 10 min per child on the part of the educator. It includes a 70-item vocabulary checklist, as well as questions concerning the child's use of decontextualized language with respect to objects and actions distant from the here and now. The test has been standardized on a total of 5097 children aged 18-34 months. Test-retest and internal consistency measures demonstrate reliability. Validation is established through correlations with age, maternal education, the Danish One-Word Picture Vocabulary Test, and the Social Emotional Assessment Measure.


A Network Approach to Compliance: A Complexity Science Understanding of How Rules Shape Behavior.
To understand how compliance develops both in everyday and corporate environments, it is crucial to understand how different mechanisms work together to shape individuals' (non)compliant behavior. Existing compliance studies typically focus on a subset of theories (i.e., rational choice theories, social theories, legitimacy theories, capacity theories, and opportunity theories) to understand how key variables from one or several of these theories shape individual compliance. The present study provides a first integrated understanding of compliance, rooted in complexity science, in which key elements from these theories are considered simultaneously, and their relations to compliance and each other are explored using network analysis. This approach is developed by analyzing online survey data (N = 562) about compliance with COVID-19 mitigation measures. Traditional regression analysis shows that elements from nearly all major compliance theories (except for social theories) are associated with compliance. The network analysis revealed groupings and interconnections of variables that did not track the existing compliance theories and point to a complexity overlooked in existing compliance research. These findings demonstrate a fundamentally different perspective on compliance, which moves away from traditional narrow, non-network approaches. Instead, they showcase a complexity science understanding of compliance, in which compliance is understood as a network of interacting variables derived from different theories that interact with compliance. This points to a new research agenda that is oriented on mapping compliance networks, and testing and modelling how regulatory and management interventions interact with each other and compliance within such networks. The online version contains supplementary material available at 10.1007/s10551-022-05128-8.


piRNA pathway and the potential processing site, the nuage, in the Drosophila germline.
The accurate transfer of genetic material in germline cells during the formation of gametes is important for the continuity of the species. However, animal germline cells face challenges from transposons, which seek to spread themselves in the genome. This review focuses on studies in Drosophila melanogaster on how the genome protects itself from such a mutational burden via a class of gonad-specific small interfering RNAs, known as piRNAs (Piwi-interacting RNAs). In addition to silencing transposons, piRNAs also regulate other processes, such as chromosome segregation, mRNA degradation and germline differentiation. Recent studies revealed two modes of piRNA processing – primary processing and secondary processing (also known as ping-pong amplification). The primary processing pathway functions in both germline and somatic cells in the Drosophila ovaries by processing precursor piRNAs into 23–29 nt piRNAs. In contrast, the secondary processing pathway functions only in the germline cells where piRNAs are amplified in a feed-forward loop and require the Piwi-family proteins Aubergine and Argonaute3. Aubergine and Argonaute3 localize to a unique structure found in animal germline cells, the nuage, which has been proposed to function as a compartmentalized site for the ping-pong cycle. The nuage and the localized proteins are well-conserved, implying the importance of the piRNA amplification loop in animal germline cells. Nuage components include various types of proteins that are known to interact both physically and genetically, and therefore appear to be assembled in a sequential order to exert their function, resulting in a macromolecular RNA-protein complex dedicated to the silencing of transposons.