Modern psychiatric classification in research and clinical practice.
After general "antipsychiatric" criticism and with empirical results, which demonstrated partly very low interrater reliabilities of conventional psychiatric diagnosis a new appreciation of nosological difficulties in psychiatry arose. Based on philosophical concepts of logical empiricism which had been established in philosophy at the beginning of our century the operational psychiatric diagnosis was developed. The first system, the Research Diagnostic Criteria (RDC), was used for scientifical purpose in the early seventies. Based upon this system the American Psychiatric Association (APA) introduced DSM-III in 1980, which had a very strong influence on psychiatric diagnosis and classification during the last ten years. Operational psychiatric diagnosis is demanding precise criteria for inclusion and exclusion and definite diagnostic rules. The tenth revision of the International Classification of Diseases (ICD-10) of the WHO is in preparation and will also contain an operational approach. This new instrument will be introduced from 1992 in WHO member states. As a result of a number of empirical studies an improvement of interrater reliability in psychiatric diagnosis using DSM-III, DSM-III-R and the drafts of ICD-10 as well was demonstrated. An other important innovation of modern classification systems is the multiaxial diagnosis. These approaches have a number of advantages for clinicians, for communication in health service systems as well as for epidemiological and biological psychiatric research.


[Difficulties in the differential diagnosis of hyponatremia presenting with severe neuropsychiatric symptoms].
Hyponatremia is the most frequent eletrolyte imbalance in hospitalized geriatric patient. The accompanying signs and symptoms can run a wide range and, therefore, these patients are usually admitted to various departments, i.e. neurology and/or traumatology first. Directed laboratory investigations demonstrate severe hyponatremia. Differential diagnosis can be very difficult and complex in the clinical settings. Firstly, spurious forms of hyponatremia have to be excluded, then the underlying cause should elucidated based on the patients hydration status and serum osmolarity. Hyponatremia can be divided into hyper-, hypo- and normovolemic forms. Moreover, it can be further classified as hypo-, iso- and hyperosmolar hyponatremias. The differentiation between renal and extrarenal salt wasting forms is hinged on the urine sodium concentration. Syndrome of inappropriate antidiuretic hormone secretion is the most common cause of normovolemic, hypoosmolar forms (named also as Schwartz-Bartter syndrome). The authors aimed to shed light on the often insurmountable difficulties of the diagnosis, differential diagnosis and appropriate treatment of this very frequent electrolyte imbalance by presenting a clinical case report. Their purported aim reflects upon the wide array of ethiopathogenesis of hyponatremia: various endocrine, renal diseases, inappropriateness of antidiuretic hormone secretion as well as the role of different medications (e.g. diuretics). This fine-tuned and intricate physiology of sodium metabolism could fortuitously be overturned by these mechanisms.


Current concepts on cleft lip and palate etiology.
Nonsyndromic cleft lip with or without cleft palate is the most common craniofacial anomaly affecting around 1 in 700 live births worldwide. Clefts of the human face can be classified anatomically as cleft palate only (CPO), cleft lip only (CLO), cleft lip and palate (CLP) or a combined group of cleft lip with or without cleft palate (CL/P), based on different in embryologic development. These malformations have some genetic origin, in fact several association studies have been performed to obtain important information about the candidate genes; but more important are gene-environment interactions that play an increasing role in its etiology. Epidemiological studies have shown how environmental factors (alcohol, smoking, drugs), as well as possible gene-environment interactions, play an important role in the onset of the malformation. On the contrary, folic acid intake seems to have a protective effect. In this review, we analyze the role of environmental factors related to onset of cleft.


Using lower eyelid fascial slings for recalcitrant burn ectropion.
Burns of the lower eyelid represent a difficult management problem. Even with skin grafting, scarring and contraction can result in ectropion. This condition creates a marked aesthetic deformity, poses a risk of corneal exposure, and jeopardizes the patient's vision. Historically, full-thickness skin grafts and tarsorrhaphies have been described to manage recurrent ectropion. The lower eyelid fascial sling (LEFS) uses a temporalis fascial strip to create a suspension sling for the lower eyelid. In a series of 7 patients, the LEFS procedure resolved the symptoms permanently in all patients. Because the LEFS procedure counteracts the natural scar contraction forces of lower eyelid burns, it can resolve recalcitrant ectropion successfully while preserving the function and aesthetics of the lower eyelid.


[Effect of morphine on pain threshold and C-fos expression induced by substance P].
Using pain measurement and immunohistochemistry, we have observed that the intrathecal injection (it) SP produced dose-related pain response. A larger dose of SP (5 micrograms, 10 micrograms) could reduce the pain threshold and evoke c-fos expression, but no obvious effect was produced by a smaller dose of SP (0.5 microgram, 1 microgram). Morphine (it) not only increased the pain threshold, but also inhibited pain response and c-fos expression of spinal cord induced by it SP, the effects could be reversed by naloxone.


Application and Feasibility Study of Integrated Nursing Information Construction in Nephrology Nursing.
With the continuous deepening of medical reforms and the continuous attempts and explorations of various management models, the traditional health care model is undergoing tremendous changes, and patients' needs for medical institutions are becoming more and more comprehensive. Medical institutions are meeting the needs of providing medical services to patients at the same time. It is even more necessary to change our thinking and enhance the service concept. This article is based on case-based deep learning hospital nursing business process reengineering and the application and feasibility study of integrated nursing information construction in nephrology nursing. This article uses the literature analysis method, the social survey method, and other methods to discuss the construction of integrated nursing information. On the one hand, the content of this article uses the concept of process reengineering to analyze the current development status and existing problems of the hospital care industry and find countermeasures to solve problems. On the other hand, the main research content of this article is the construction of integrated nursing information and its analysis of the application and feasibility of nursing in the nephrology department. At the same time, under the background of the rapid development of the mobile Internet, we will carry out extended thinking on the continuous transformation of the construction of nursing information. According to the survey results, 87.5% of patients in the nephrology department are dissatisfied with the current hospital's work efficiency, and 85.7% of the nursing staff in the nephrology department are generally satisfied with the information management of the current department. After the implementation of the hospital information integration system, patient satisfaction is as high as 98.2%, and the satisfaction of medical staff reached 94.2%. The construction of integrated nursing information has played a great role in the application of nephrology nursing.


A novel one-dimensional zinc coordination polymer containing 4-methylbenzoate and 4,4-bipyridine.
The title compound, catena-poly[[tris(mu-4-methylbenzoato)-kappa(2)O:O;kappa(4)O:O'-(4-methylbenzoato-kappa(2)O,O')dizinc(II)]-mu-4,4'-bipyridine-kappa(2)N:N'], [Zn(2)(C(8)H(7)O(2))(4)(C(10)H(8)N(2))](n), is a novel coordination polymer. The asymmetric unit contains two unique Zn(II) ions, four 4-methylbenzoate ligands and one 4,4'-bipyridine (4,4'-bpy) ligand, all in general positions. The four 4-methylbenzoate ligands link the two Zn(II) centres to form a dinuclear unit, with a Zn...Zn separation of 3.188 (2) A, which can be regarded as a supramolecular secondary building unit (SBU). These SBUs are further bridged by 4,4'-bpy ligands, forming a novel one-dimensional infinite chain. There are pi-pi stacking interactions between the benzene rings of the 4-methylbenzoate ligands and the pyridyl rings of the 4,4'-bpy ligands, leading to the formation of a corrugated layer. These layers are further assembled via C-H...O hydrogen bonds into a three-dimensional supramolecular network structure. Coordination polymers such as the title compound are of interest for their potential applications as functional materials.


Ofloxacin and the treatment of pulmonary disease due to Mycobacterium fortuitum.
A 58-year-old man with pulmonary disease due to Mycobacterium fortuitum was treated successfully with ofloxacin (0.3 g daily). Negative conversion of sputum culture was achieved immediately after administration of ofloxacin, and the chest roentgenogram also improved.


Do junior doctors make more prescribing errors than experienced doctors when prescribing electronically using a computerised physician order entry system combined with a clinical decision support system? A cross-sectional study.
Prescribing errors can lead to inconvenience, morbidity and mortality. It is therefore crucial to educate doctors to prescribe safely, efficiently and effectively. To create an effective educational programme, it is essential to understand which errors are made and by whom. The aim of this study is to explore if the experience level of the doctor influences how many and which prescribing errors are made in a European academic teaching hospital, where a computerised physician order entry system (CPOE) with a clinical decision support system (CDSS) is exclusively used. Prescriptions for all inpatients in an academic teaching hospital were collected in June 2021. All prescriptions with an alert generated by the CDSS which could not be handled by a pharmacy technician according to local protocol were checked for errors. Identified errors were categorised by type and severity. A total of 130 538 prescriptions were newly made or altered by doctors. Of these prescriptions, 1914 (1.5%) were retained for a check by the pharmacist. These contained 430 prescribing errors (0.3% of total prescriptions). Doctors not in specialty training and those in specialty training made more prescribing errors than consultants (0.5% and 0.5% vs 0.1%; p<0.001). Doctors in specialty training made relatively more drug-drug interaction errors than consultants (n=31 (16%) vs n=3 (3%), p<0.05). No significant difference was found regarding the severity of the errors. Doctors not in specialty training and doctors in specialty training, who are the less experienced doctors, make more prescribing errors than consultants, even with the use of a CPOE combined with CDSS. The type of errors differ between doctors of different experience levels. This finding provides a solid basis for specific additional education to medical students, doctors not in specialty training and doctors in specialty training.


A Framework for the Joint Placement of Edge Service Infrastructure and User Plane Functions for 5G.
Achieving less than 1 ms end-to-end communication latency, required for certain 5G services and use cases, is imposing severe technical challenges for the deployment of next-generation networks. To achieve such an ambitious goal, the service infrastructure and User Plane Function (UPF) placement at the network edge, is mandatory. However, this solution implies a substantial increase in deployment and operational costs. To cost-effectively solve this joint placement problem, this paper introduces a framework to jointly address the placement of edge nodes (ENs) and UPFs. Our framework proposal relies on Integer Linear Programming (ILP) and heuristic solutions. The main objective is to determine the ENs and UPFs' optimal number and locations to minimize overall costs while satisfying the service requirements. To this aim, several parameters and factors are considered, such as capacity, latency, costs and site restrictions. The proposed solutions are evaluated based on different metrics and the obtained results showcase over 20 % cost savings for the service infrastructure deployment. Moreover, the gap between the UPF placement heuristic and the optimal solution is equal to only one UPF in the worst cases, and a computation time reduction of over 35 % is achieved in all the use cases studied.


[Metabolism and biological function of cardiolipin].
Cardiolipin (CL) is a phospholipid, which is exclusively located in mitochondria, and has a unique structure that consists of 2 phosphate residues and 4 kinds of fatty acyl chains. Cardiolipin plays an important role in regulating various kinds of mitochondrial proteins such as electron transport complexes, carrier proteins and phosphate kinases, and is also essential for the organization of particular mitochondrial structures such as cristae and contact sites. Mitochondrial phospholipase D hydrolyzes CL to produce phosphatidic acid, which is required for mitochondrial fusion. Oxidative stress-induced peroxidation of CL occurs because CL is rich in polyunsaturated fatty acids, especially linoleic acid. Accumulation of CL hydroperoxide (CLOOH) triggers the initiation of apoptosis. Formation of CLOOH causes the release of proapoptotic factors such as cytochrome c from the inner mitochondrial membrane and triggers opening of the permeability transition pore. Levels of CL decrease in the heart following ischemia or disease. Apoptosis is enhanced in temperature-dependent mutant cells whose amounts of CL reduce to half when compared to that of wild type cells. Low levels of CL cause the accumulation of CLOOH and enhance sensitivity to apoptosis. Accumulation of CLOOH in mitochondria causes instability of the membrane, because swelling of mitochondria is induced by the presence of CLOOH in the membrane and is significantly enhanced in CLOOH-loaded mitochondria by the addition of inducer of swelling.


The human selenoproteome: recent insights into functions and regulation.
Selenium (Se) is a nutritional trace mineral essential for various aspects of human health that exerts its effects mainly through its incorporation into selenoproteins as the amino acid, selenocysteine. Twenty-five selenoprotein genes have been identified in humans and several selenoproteins are broadly classified as antioxidant enzymes. As progress is made on characterizing the individual members of this protein family, however, it is becoming clear that their properties and functions are quite diverse. This review summarizes recent insights into properties of individual selenoproteins such as tissue distribution, subcellular localization, and regulation of expression. Also discussed are potential roles the different selenoproteins play in human health and disease.


Validation of the Impact of Event Scale With Modifications for COVID-19 (IES-COVID19).
Viral outbreaks can be experienced as disruptive and can be associated with trauma-related stress symptoms. In the current study, we adjusted the Dutch version of the Impact of Event Scale (IES) to assess traumatic stress symptoms related to the impact of the COVID-19 outbreak. The psychometric properties of this Impact of Event Scale with modifications for COVID-19 (IES-COVID19) were investigated by administering the IES-COVID19 to 380 university students who participated during the early stage of the COVID-19 outbreak, upon invitation via e-mail. Using confirmatory factor analysis, the factor structure of the IES-COVID19 was found to be similar to the original IES, indicating two latent factors: intrusion and avoidance, χ2 (85) = 147.51, CFI = .92, TLI = .90, RMSEA = .044, SRMR = .049. Cronbach's alpha showed acceptable internal consistency of the total IES-COVID19, α = .75. Pearson's correlations of the IES-COVID19 over time were also sufficient, demonstrating adequate test-retest reliability, r = .62. Significant correlations (ranging between .15 and .50) between the IES-COVID19 and symptoms of depression, anxiety, stress, stress-related rumination, as well as negative social interactions, demonstrate adequate convergent validity. Overall, the IES-COVID19 shows to be a valid and reliable measure that can be utilized to investigate trauma-related stress symptoms of intrusion and avoidance related to the short- and long-term impact of the COVID-19 outbreak.


Applications of carbon nanotubes in the twenty-first century.
Carbon nanotubes, which consist of rolled graphene sheets built from sp(2) hybridized carbon atoms, are now attracting scientists from various disciplines due to their fascinating physico-chemical properties. In this account, we will review the recent progress on the synthetic techniques for the large-scale production of carbon nanotubes, especially focusing on the floating-catalyst method used in the chemical vapour deposition (CVD) process. We will also describe effective purification methods avoiding structural damage, and discuss the electrochemical applications of these systems including the fabrication of: (i) lithium-ion secondary batteries; (ii) lead-acid batteries; (iii) electric double-layer capacitors; (iv) fuel cells; and (v) multifunctional fillers in polymer composites. We foresee that carbon nanotubes will find numerous applications and take an important place in the development of emerging technologies in the near future.


Development of a new mini-invasive tumour hyperthermia probe using high-temperature water vapour.
A new mini-invasive hyperthermia probe using high-temperature water vapour for deep regional tumour treatment was developed in this paper. The vacuum insulation mechanism was incorporated into the probe to avoid heating damage to the normal tissues around the edge of the insertion path. To better understand the heat transfer behaviour in living tissues due to operation of the probe, theoretical models based on the Pennes' equation were established and two closed form analytical solutions under constant flux or temperature heating at the tip of probe were obtained. Parametric studies were performed to investigate the influence of various parameters on the temperature response of tissues heated by the probe. Further, several simulating experiments on the actual heating performance of the probe fabricated in this paper were conducted on the in vitro biological materials (fresh pork) and phantom gel. It was demonstrated that the probe can cause a high enough temperature over the treatment area to thermally destroy the tumour tissue in due time, while the temperature over the surrounding healthy tissues can be kept below a safe threshold value. This mini-invasive heating probe may have significant applications in future clinical tumour hyperthermia.


Getting started in research: research questions, supervisors and literature reviews.
Research can seem daunting, especially for trainees and early career researchers. This paper focuses on how to formulate and begin a research project such as the RANZCP Scholarly Project. We outline an approach to framing a research question, developing theses and hypotheses, choosing a supervisor and conducting a literature review. Through systematic planning early career researchers and other clinicians can plan and conduct research suitable for the Scholarly Project or other research activity.


Targeting Neuropilin-1 Suppresses the Stability of CD4+ CD25+ Regulatory T Cells via the NF-κB Signaling Pathway in Sepsis.
Neuropilin-1 (Nrp-1) contributes to maintaining the stability of CD4+ CD25+ regulatory T cells (Tregs). We investigated the impact of Nrp-1 on the stability of CD4+ CD25+ Tregs, and the underlying signaling pathways, in a model of sepsis. Splenic CD4+ CD25+ Tregs were either treated with anti-Nrp-1, transfected to silence Nrp-1 and inhibitor of NF-κB kinase subunit beta (IKKβ), or administered ammonium pyrrolidine dithiocarbamate (PDTC), followed by recombinant semaphorin 3A (rSema3A), in a simulation of sepsis. After the creation of a sepsis model in mice, anti-Nrp-1 was administered. The expression of the gene encoding forkhead box protein P-3 foxp3-Treg-specific demethylated region (foxp3-TSDR), the apoptosis rate, the expression of Foxp-3, cytotoxic T-lymphocyte-associated protein-4 (CTLA-4), and transforming growth factor β1 (TGF-β1), interleukin 10 (IL-10) and TGF-β1 secretion, and the NF-κB signaling activity of CD4+ CD25+ Tregs were determined. Sepsis simulation with or without rSema3A increased the stability of CD4+ CD25+ Tregs, including an increase in the expression of Foxp-3, CTLA-4, and TGF-β1, decreases in apoptosis and the methylation of foxp3-TSDR, increases in the secretion of TGF-β1 and IL-10, and an increase in the immunosuppressive effect on CD4+ T lymphocytes. Silencing of Nrp-1 or anti-Nrp-1 treatment abrogated lipopolysaccharide (LPS) stimulation with or without an rSema3A-mediated effect. Sepsis simulation increased the DNA-binding activity of NF-κB, as well as the ratios of phosphorylated IKKβ (p-IKKβ) to IKKβ and p-P65 to P65 in vitro and vivo Silencing of IKKβ expression or PDTC treatment suppressed the stability of CD4+ CD25+ Tregs in LPS-induced sepsis. Weakening Nrp-1 reduced the stability of CD4+ CD25+ Tregs by regulating the NF-κB signaling pathway; thus, Nrp-1 could be a new target for immunoregulation in sepsis.


Modular 3-cm uniportal video-assisted thoracoscopic left upper lobectomy with systemic lymphadenectomy.
Uniportal video-assisted thoracoscopic lobectomy for non-small-cell lung cancer is accepted worldwide, with incisions ranging from 4 to 6 cm. We believed in less invasive and more precise that uniportal video-assisted thoracoscopic lobectomy could be. Therefore, we performed modular uniportal thoracoscopic lobectomy with systemic lymphadenectomy on left upper lobe using a 3-cm-diameter port. And the modular surgical route was arranged in seven modules. Anesthesia, patient positioning and instruments play an important role in the surgery. From October 2014 to June 2015, 96 patients underwent this modular surgery and all patient were discharged uneventfully with no postoperative deaths. Compared with multi-port VATS, the operation time were longer than multiport video-assisted thoracoscopic surgery (VATS) (164.70±12.50 vs. 160.70±11.60 min, P>0.05), and the mean lymphadenectomy station was 6.00±0.77, and the mean lymphadenectomy number was 17.58±5.33. There is no significant difference on lymphadenectomy. Thus, modular uniportal video-assisted thoracoscopic lobectomy with systemic lymphadenectomy on left upper lobe using a 3-cm-diameter port is a safe, feasible, and less painful technique for select patients with lung disease.


Efficacy of angiotensin receptor neprilysin inhibitor in Asian patients with refractory hypertension.
Sacubitril/valsartan, simultaneously inhibits neprilysin and angiotensin II receptor, showed an effect in reducing blood pressure (BP). The authors aimed to study whether it can be used as an antihypertensive agent in patients with refractory hypertension who have already been treated. A total of 66 Chinese patients with refractory hypertension were enrolled. Patients received sacubitril/valsartan 200 instead of angiotensin II receptor blocker or angiotensin converting enzyme inhibitor while other agents continued. If BP was uncontrolled after 4 weeks, sacubitril/valsartan was increased to 400 mg. The BP reduction was evaluated by office BP and ambulatory BP monitoring after 8-week treatment. The baseline office BP and mean arterial pressure (MAP) were 150.0/95.0 mmHg and 113.3 mmHg. BP and MAP reduced to 130.6/83.2 mmHg and 99.0 mmHg at week 8. Office BP and MAP reductions were 19.4/11.8 mmHg and 14.3 mmHg at endpoint (all p < .001). The 24-h, daytime and nighttime ambulatory BP were 146.2/89.1, 148.1/90.3, and 137.5/83.7 mmHg, respectively at baseline, and BP reduced to 129.6/79.8, 130.6/81.1, and 121.7/75.8 mmHg, respectively at week 8. The 24-h, daytime and nighttime ambulatory BP reductions were 16.6/9.3, 17.5/9.2, and 15.8/7.9 mmHg, respectively at endpoint (all p < .001). Sacubitril/valsartan significantly reduced office and ambulatory BP in refractory hypertension patients. Our study provided new evidence for sacubitril/valsartan in refractory hypertension.


l-Canavanine Transport and Utilization in Developing Jack Bean, Canavalia ensiformis (L.) DC. [Leguminosae].
l-Canavanine, the guanidinooxy structural analog of l-arginine, is an important nonprotein amino acid of many leguminous plants with nitrogen storage a major proported role. l-[Guanidinooxy-(14)C]canavanine, [(14)C] urea, and [(15)N]urea were injected separately into the fleshy, green cotyledons of 9-day old jack bean plants, Canavalia ensiformis (L.) DC. [Leguminosae]. There was significant transport of canavanine from the cotyledons to the aboveground portions of the plant, but not to the roots. Within 1.5 hours of isotope administration, the remaining labeled canavanine was divided equally between the cotyledons and the aboveground portions of the plant. During the 48-hour postinjection period, the contribution of l-[guanidinooxy-(14)C]canavanine to the total (14)carbon of the cotyledons decreased rapidly while it increased in the aboveground portions of the plant.[(14)C]Urea is degraded very rapidly; only 4.4% of the initial dose remained after 1.5 hours. Urea is catabolized so effectively within the cotyledons that not even 2% of the administered urea can be detected in tissues outside of these storage organs. [(15)N]Urea supplied to the developing coytledons leads to rapid (15)N incorporation into the amino nitrogen of glutamic acid and/or glutamine (28% (15)N abundance after 3 hours). Other amino acids are labeled but less heavily. The data are consistent with the proported role for l-canavanine of nitrogen storage within the developing cotyledons and cotyledonary canavanine is transported very effectively to the aboveground portions of the plant. It is not yet clear how efficiently this transported canavanine supports the nitrogen metabolism of the developing plant.


Tuberculosis, the culprit behind nonhealing oral lesions: report of two cases.
Nonhealing painless lesions of long duration that did not respond to conventional therapy are the diagnostic challenge to clinician. Two such atypical cases of nonhealing oral lesions manifesting as gingival swelling and odontogenic cyst diagnosed histopathologically as primary oral tuberculosis are presented here. The purpose of this presentation is to point out that, in the absence of any apparent systemic infection the oral tuberculous lesions in the mouth may be discovered before the diagnosis of systemic tuberculosis and the histopathological finding of a persistent nonhealing oral lesion is an important aid in arriving at a diagnosis of this underlying condition.


Outcomes of stereotactic body radiotherapy for unresectable primary or recurrent cholangiocarcinoma.
To report the results of stereotactic body radiotherapy (SBRT) for unresectable primary or recurrent cholangiocarcinoma. From January 2005 through August 2013, 58 patients with unresectable primary (n = 28) or recurrent (n = 30) cholangiocarcinoma treated by SBRT were retrospectively analyzed. The median prescribed dose was 45 Gy in 3 fractions (range, 15 to 60 Gy in 1-5 fractions). Patients were treated by SBRT only (n = 53) or EBRT + SBRT boost (n = 5). The median tumor volume was 40 mL (range, 5 to 1,287 mL). The median follow-up duration was 10 months (range, 1 to 97 months). The 1-year, 2-year overall survival rates, and median survival were 45%, 20%, and 10 months, respectively. The median survival for primary group and recurrent group were 5 and 13 months, respectively. Local control rate at 1-year and 2-year were 85% and 72%, respectively. Disease progression-free survival rates at 1-year and 2-year were 26% and 23%, respectively. In univariate analysis, ECOG performance score (0-1 vs. 2-3), treatment volume (<50 vs. ≥50 mL), and pre-SBRT CEA level (<5 vs. ≥5 ng/mL) were significant in overall survival rate. In multivariate analysis, ECOG score (p = 0.037) and tumor volume (p = 0.030) were statistically significant. In the recurrent tumor group, patients with >12 months interval from surgery to recurrence showed statistically significant higher overall survival rate than those with ≤12 months (p = 0.026). Six patients (10%) experienced ≥grade 3 complications. SBRT can be considered as an effective local modality for unresectable primary or recurrent cholangiocarcinoma.


Cascade-Targeting Poly(amino acid) Nanoparticles Eliminate Intracellular Bacteria via On-Site Antibiotic Delivery.
Intracellular bacteria in latent or dormant states tolerate high-dose antibiotics. Fighting against these opportunistic bacteria has been a long-standing challenge. Herein, the design of a cascade-targeting drug delivery system (DDS) that can sequentially target macrophages and intracellular bacteria, exhibiting on-site drug delivery, is reported. The DDS is fabricated by encapsulating rifampicin (Rif) into mannose-decorated poly(α-N-acryloyl-phenylalanine)-block-poly(β-N-acryloyl-d-aminoalanine) nanoparticles, denoted as Rif@FAM NPs. The mannose units on Rif@FAM NPs guide the initial macrophage-specific uptake and intracellular accumulation. After the uptake, the detachment of mannose in acidic phagolysosome via Schiff base cleavage exposes the d-aminoalanine moieties, which subsequently steer the NPs to escape from lysosomes and target intracellular bacteria through peptidoglycan-specific binding, as evidenced by the in situ/ex situ co-localization using confocal, flow cytometry, and transmission electron microscopy. Through the on-site Rif delivery, Rif@FAM NPs show superior in vitro and in vivo elimination efficiency than the control groups of free Rif or the DDSs lacking the macrophages- or bacteria-targeting moieties. Furthermore, Rif@FAM NPs remodel the innate immune response of the infected macrophages by upregulating M1/M2 polarization, resulting in a reinforced antibacterial capacity. Therefore, this biocompatible DDS enabling macrophages and bacteria targeting in a cascade manner provides a new outlook for the therapy of intracellular pathogen infection.


Microbiomes and metabolomes of dominant coral reef primary producers illustrate a potential role for immunolipids in marine symbioses.
The dominant benthic primary producers in coral reef ecosystems are complex holobionts with diverse microbiomes and metabolomes. In this study, we characterize the tissue metabolomes and microbiomes of corals, macroalgae, and crustose coralline algae via an intensive, replicated synoptic survey of a single coral reef system (Waimea Bay, O'ahu, Hawaii) and use these results to define associations between microbial taxa and metabolites specific to different hosts. Our results quantify and constrain the degree of host specificity of tissue metabolomes and microbiomes at both phylum and genus level. Both microbiome and metabolomes were distinct between calcifiers (corals and CCA) and erect macroalgae. Moreover, our multi-omics investigations highlight common lipid-based immune response pathways across host organisms. In addition, we observed strong covariation among several specific microbial taxa and metabolite classes, suggesting new metabolic roles of symbiosis to further explore.


Diagnostic value of processing cytologic aspirates of renal tumors in agar cell (tissue) blocks.
To adapt a method enabling utilization of most of the harvest from a fine needle aspirate in an effort to improve diagnostic accuracy in the assessment of a renal tumor in a single histologic slide. In a series of 43 renal tumors, 2 fine needle aspirations were performed, 4 smears were prepared after each aspiration for conventional cytology and the remaining aspirate was processed for the improved agar microbiopsy (AM) method. Conventional cytology slides, AM slides and surgical specimens were diagnosed separately, after which the diagnoses were compared. Immunohistochemistry was performed as required on the AM sections. Surgical specimens served as the gold standard. In 53% of conventional cytologic smears, the cellular yield was sufficient to render a correct diagnosis. In 12% the diagnosis was incorrect, in 21% only a differential diagnosis could be formulated, and in 14% too few diagnostic cells were present in the conventional smears for a cytologic diagnosis. It was, however, possible to correctly diagnose histologic sections from 97% of AM tissue blocks. In 11 cases this was facilitated with immunohistochemistry. In only 1 case did the AM tissue block contain too few cells to formulate a diagnosis; the conventional cytologic sample in this case contained enough diagnostic cells. In all cases the AM diagnosis was confirmed in the definitive surgical specimen. Our AM technique for processing fine needle aspirates from renal tumors results in a major enhancement of diagnostic accuracy of such aspirates and should be valuable in the preoperative diagnosis of large, as well as small, renal tumors.


Intracoronary macrothrombus formation during percutaneous coronary intervention despite optimal activated clotting time using bivalirudin--a case report.
The occurrence of intracoronary thrombus during percutaneous coronary intervention (PCI) is a well-known complication. It has been estimated that it complicates approximately 6% of all coronary procedures. Patients at highest risk for this complication include those with acute ischemic syndromes or with angiographically apparent thrombus. Since the development of PCI, intravenous unfractionated heparin (UFH) has remained the primary antithrombotic therapy for the prevention of periprocedural ischemic complications. The availability of a rapid "point of care'' test for dose individualization (the activated clotting time [ACT]) has facilitated this process. Other forms of antithrombotic therapies such as direct thrombin inhibitors or low-molecular-weight heparin have been proposed as more effective anticoagulants during PCI. Bivalirudin is a direct thrombin inhibitor proven to decrease post-PCI ischemic complication rate compared with UFH and have a lower vascular complication rate compared with glycoprotein IIb/IIIa receptor antagonists. We herein report a case of acute macrothrombus formation during PCI despite adequate ACT achieved with bivalirudin.


Numerically "exact" simulations of a quantum Carnot cycle: Analysis using thermodynamic work diagrams.
We investigate the efficiency of a quantum Carnot engine based on open quantum dynamics theory. The model includes time-dependent external fields for the subsystems controlling the isothermal and isentropic processes and for the system-bath (SB) interactions controlling the transition between these processes. Numerical simulations are conducted in a nonperturbative and non-Markovian SB coupling regime by using the hierarchical equations of motion under these fields at different cycle frequencies. The work applied to the total system and the heat exchanged with the baths are rigorously evaluated. In addition, by regarding quasi-static work as free energy, we compute the quantum thermodynamic variables and analyze the simulation results by using thermodynamic work diagrams for the first time. Analysis of these diagrams indicates that, in the strong SB coupling region, the fields for the SB interactions are major sources of work, while in other regions, the field for the subsystem is a source of work. We find that the maximum efficiency is achieved in the quasi-static case and is determined solely by the bath temperatures, regardless of the SB coupling strength, which is a numerical manifestation of Carnot's theorem.


Development and Pilot Testing of a Patient-Centered Web-Based Reproductive Decision Support Tool for Primary Care.
Patient-centered counseling to help women achieve their reproductive goals is an essential yet often absent component of primary care. We developed and piloted MyPath, a novel web-based decision support tool integrating reproductive goals assessment, information about optimizing health before pregnancy, and contraceptive decision support, for use prior to primary care visits in the Veterans Administration (VA). We created MyPath using best practices for decision tool development, including a conceptual framework informed by theory and user-centered design with input from patients, providers, and scientific experts. We conducted a non-randomized pilot in two VA Women's Health primary care clinics. A control group (n = 28) was recruited prior to and intervention group (n = 30) recruited after introduction of MyPath into clinics. Women Veterans ages 18-44 with an upcoming visit scheduled with one of eight providers. After recruitment of controls, providers and staff received a brief introduction to MyPath. Patients scheduled to see providers in the intervention phase used MyPath on an iPad in the waiting room prior to their visit. Acceptability, feasibility, discussions about pregnancy and/or contraceptive needs, and contraceptive decision quality by a survey of participants and providers. Nearly all participants who used MyPath reported they learned new information (97%) and would recommend it to other Veterans (93%). No providers reported that MyPath significantly increased workload. A greater proportion of intervention participants reported having discussions about reproductive needs in their visit compared to controls (93% vs 68%; p = 0.02). Intervention participants also experienced greater increases in pre-/post-visit knowledge and communication self-efficacy and a trend towards greater reduction in contraceptive decision conflict compared to controls. MyPath was highly acceptable to women, increased the proportion of primary care visits addressing reproductive needs, and improved decision quality without increasing providers' perceived workload. A larger randomized evaluation of effectiveness is warranted.


Effects of chronic exposure to crack cocaine on the respiratory tract of mice.
Smoked cocaine (crack cocaine) causes several forms of injury to the respiratory tract, including asthma exacerbations, lung edema and hemorrhage, and nasal mucosal alterations. Few studies, however, have assessed respiratory tract pathology in habitual users of crack cocaine. Here, we describe the histological alterations in the respiratory tract of mice caused by chronic inhalation of crack cocaine. Twenty 2-month-old BALB/c mice were exposed to the smoke of 5 g crack cocaine in an inhalation chamber once a day for two months and compared to controls (n = 10). We then morphometrically analyzed nose and bronchiolar epithelial alterations, bronchiolar and alveolar macrophage cell density, alveolar hemosiderin content, and in addition determined the vasoconstriction index and the wall thickness of pulmonary arteries. The serum cocaine level was 212.5 ng/mL after a single inhalation. The mucus content of the nasal epithelium increased in crack-exposed animals, and the nasal and bronchial epithelium thickness decreased significantly. The alveolar hemosiderin content and the alveolar and bronchiolar macrophage cell density increased in animals exposed to crack. The vasoconstriction index increased in the pulmonary arteries of the exposed group. Chronic crack cocaine inhalation causes extensive histological changes along the entire respiratory tract.


Controlled Thermoreversible Formation of Supramolecular Hydrogels Based on Poly(vinyl alcohol) and Natural Phenolic Compounds.
Supramolecular hydrogels have promising applications in a wide variety of fields including 3D bioprinting, sensors and actuators, biomedicine, and controlled drug delivery. This communication reports the facile reversible thermotriggered formation of novel pH-responsive supramolecular hydrogels based on poly(vinyl alcohol) (PVA) bonded via dynamic H-bridge with small phenolic biomolecules. PVA and phenolic compounds form a clear solution when they are physically mixed in water at high temperature, but a fast gelation is produced at room temperature through multiple strong H-bonding interactions. The structure and type of functional groups of different phenolic molecules allow preparing hydrogels with tailor-made viscoelastic properties, controlled low phase transition temperature, and pH-dependent swelling behavior. This combination makes these supramolecular networks very interesting candidates to be used in 3D bioprinting and topical drug delivery of thermolabile biomolecules.


Oral oncolytic treatment for chronic lymphocytic leukemia.
The objective of this article is to review the current supporting literature for the use of oral oncolytics in chronic lymphocytic leukemia, consideration for their use and management of adverse drug events that may limit use. NCCN guidelines were utilized to determine available oral options for treatment of chronic lymphocytic leukemia. A literature review was carried out through PubMed to find relevant clinical trials that evaluated the efficacy and safety of Bruton's tyrosine kinase inhibitors, BCL-2 inhibitors and PI3K-δ inhibitors. Medication package inserts and primary literature regarding toxicity were used to determine appropriate adverse drug event management. A total of 7 clinical trials were found for the evaluation the efficacy and safety of burton tyrosine kinase inhibitor, 1 clinical trial for the BCL-1 inhibitor venetoclax and 4 trials were for PI3K-δ inhibitors. The data from these studies suggest that ibrutinib can be used first line in previously untreated patients and relapsed/refractory patients as well as acalabrutinib. The data also support the use of venetoclax, idelalisib, and duvelisib in relapsed/refractory chronic lymphocytic leukemia patients. The use of oral-only oncolytics could be a viable option for reducing the risk of infection due to limiting exposure to healthcare settings. Current literature suggests oral oncolytics may be an option, but there are several considerations to evaluate including medication adherence, drug-drug interactions, adverse events, and financial toxicity.


Genome-wide transcript expression analysis in the uterovaginal junction in association with fertile period in Tsaiya ducks.
We performed the first genome-wide expression analysis to compare the differences in gene expression in the female sperm reservoir of the duck reproductive tract between two groups with long and short fertile periods to identify factors that may be associated with the fertile period using an oligonucleotide microarray. RNA was extracted from the uterovaginal junction (UV junction) of the two groups. Affymetrix chips containing comprehensive coverage of 32773 transcripts were hybridized with biotin-labeled cRNA, and three biological repeats were performed. We identified 27 transcripts as being differentially regulated. Interestingly, by mapping the differentially expressed transcripts to annotated pathways, we found that Neuropeptide Y (NPY), the RNA expression of which was increased by 2.96-fold in the short-fertile-period group as compared with the long-fertile-period group in our experiment, has been shown to reduce blood flow and substance supply to local tissues. Enah/Vasp-like (EVL), the RNA expression of which was significantly increased by 1.77-fold in the short-fertile-period group as compared with the long-period group, has been demonstrated to be important in activated T-cells. In contrast, trafficking kinesin-binding protein 1 (TRAK1), the expression of which was increased by 2.33-fold in the long-period group as compared with its counterparts, has been suggested to inhibit precocious activation of sperm and prolong sperm life in the female sperm reservoir. The results of real-time PCR confirmed the data obtained by microarray analysis. Our study demonstrated that combining global gene expression investigation with annotated pathway resources contributes to the understanding of sperm life when sustained in the UV junction.


Advantage 24 nonoxynol-9 product discussed at meeting. Contraception (spermicides).
Columbia Laboratories, Inc., Miami, Florida, reported safety data on the spermicidal product Advantage 24 that show it is safe even when administered four times a day, according to data from the UN Global Program on HIV/AIDS (UNAIDS) and the US National Institutes of Health (NIH). The data were reported at a meeting of the Nonprescription Drugs, Reproductive Health Drugs, Anti-Infective Drugs, and Antiviral Drugs advisory committees of the US Food and Drug Administration. Because of its favorable safety profile, Advantage 24 was the only product that UNAIDS selected for testing to determine its effectiveness against the spread of sexually transmitted diseases (STDs). Therefore, in May 1996, Columbia Laboratories and UNAIDS announced the initiation of an international Phase III clinical study of Advantage 24 for the prevention of the heterosexual transmission of HIV and STDs in approximately 2000 women. Additionally, the NIH is performing similar testing in Africa. Advantage 24, which uses Columbia Laboratories' patented bioadhesive delivery system to deliver nonoxynol-9, currently is marketed in the US by Lake Pharmaceuticals, Inc., and in Canada by Roberts Pharmaceuticals, as a long-lasting, vaginally delivered spermicide, pursuant to strategic alliance agreements with Columbia Laboratories.


Calcium-channel blocker withdrawal in a pregnant woman.
We present a case report of a woman for whom a topical mucosal ulcer drug was prescribed and the pharmacist erroneously dispensed a calcium channel blocker (CCB), resulting in toxicities from the drug and withdrawal symptoms when attempts were made to stop the CCB. The pharmacology and toxicology of CCBs are discussed, particularly in relation to the adverse experiences of the case.


Redox Metabolism-Associated Molecular Classification of Clear Cell Renal Cell Carcinoma.
Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma. Redox metabolism has been recognized as the hallmark of cancer. But the concrete role of redox-related genes in patient stratification of ccRCC remains unknown. Herein, we aimed to characterize the molecular features of ccRCC based on the redox gene expression profiles from The Cancer Genome Atlas. Differentially expressed redox genes (DERGs) and vital genes in metabolism regulation were identified and analyzed in the ccRCC. Consensus clustering was performed to divide patients into three clusters (C1, C2, and C3) based on 139 redox genes with median FPKM value > 1. We analyzed the correlation of clusters with clinicopathological characteristics, immune infiltration, gene mutation, and response to immunotherapy. Subclass C1 was metabolic active with moderate prognosis and associated with glucose, lipid, and protein metabolism. C2 had intermediate metabolic activity with worse prognosis and correlated with more tumor mutation burden, neoantigen, and aneuploidy, indicating possible drug sensitivities towards immune checkpoint inhibitors. Metabolic exhausted subtype C3 showed high cytolytic activity score, suggesting better prognosis than C1 and C2. Moreover, the qRT-PCR was performed to verify the expression of downregulated DERGs including ALDH6A1, ALDH1L1, GLRX5, ALDH1A3, and GSTM3, and upregulated SHMT1 in ccRCC. Overall, our study provides an insight into the characteristics of molecular classification of ccRCC patients based on redox genes, thereby deepening the understanding of heterogeneity of ccRCC and allowing prediction of prognosis of ccRCC patients.


Scavengers of reactive oxygen species, paracalcitol, RhoA, and Rac-1 inhibitors and tacrolimus inhibit angiotensin II-induced actions on glomerular permeability.
Systemic infusions of ANG II rapidly induce large, dynamic increases in the permeability of the glomerular filtration barrier (GFB) in rats. After binding to its receptor(s), ANG II generates reactive oxygen species (ROS) and produces Ca²⁺ influx into cells, leading to activation of a plethora of signaling cascades, including, e.g., calcineurin and small GTPases, such as Rac-1 and RhoA. In the present study we sought to interact with some of these cascades to test potential novel antiproteinuric agents. In anesthetized Wistar rats, the left urether was cannulated for urine collection, and blood access was achieved. Rats were infused with ANG II (16 ng·kg⁻¹·min⁻¹) alone, or together with the ROS scavengers tempol or dimethylthiourea (DMTU) or the D-vitamin analog paracalcitol, the RhoA-kinase inhibitor Y-27632, the Rac-1 inhibitor NSC-23766, or the calcineurin inhibitor tacrolimus. FITC-Ficoll-70/400 (mol.radius 10-80 Å) and ⁵¹Cr-EDTA were infused throughout the experiment. Plasma and urine samples were taken during baseline and at 5 and 15 min after the start of the infusions and analyzed by high-performance size-exclusion chromatography for determination of glomerular sieving coefficients (θ) for Ficoll10-80Å. ANG II infusion into rats caused marked increases in glomerular permeability to large Ficoll molecules (Ficoll50-80Å), which were abrogated by the ROS scavenger tempol and partly by DMTU. Paracalcitol, RhoA, and Rac-1 inhibition, and, to some extent tacrolimus, but not prostacyclin, could also inhibit the glomerular permeability actions of ANG II. Our data suggest that cellular ROS generation and active Ca²⁺ signaling are involved in ANG II-induced increases in glomerular permeability.


A common variant of PNPLA3 (p.I148M) is not associated with alcoholic chronic pancreatitis.
Chronic pancreatitis (CP) is an inflammatory disease that in some patients leads to exocrine and endocrine dysfunction. In industrialized countries the most common aetiology is chronic alcohol abuse. Descriptions of associated genetic alterations in alcoholic CP are rare. However, a common PNPLA3 variant (p.I148M) is associated with the development of alcoholic liver cirrhosis (ALC). Since, alcoholic CP and ALC share the same aetiology PNPLA3 variant (p.I148M) possibly influences the development of alcoholic CP. Using melting curve analysis we genotyped the variant in 1510 patients with pancreatitis or liver disease (961 German and Dutch alcoholic CP patients, 414 German patients with idiopathic or hereditary CP, and 135 patients with ALC). In addition, we included in total 2781 healthy controls in the study. The previously published overrepresentation of GG-genotype was replicated in our cohort of ALC (p-value <0.0001, OR 2.3, 95% CI 1.6-3.3). Distributions of genotype and allele frequencies of the p.I148M variant were comparable in patients with alcoholic CP, idiopathic and hereditary CP and in healthy controls. The absence of an association of PNPLA3 p.I148M with alcoholic CP seems not to point to a common pathway in the development of alcoholic CP and alcoholic liver cirrhosis.


Juvenile sexual offending: one-year period prevalence study within Oxfordshire.
An epidemiological survey using a postal questionnaire revealed a 1-year period prevalence rate of 1.5 official juvenile sexual offenders per 1,000 males aged 12-17 within Oxfordshire (.85 per 1,000 12-17 year olds overall). The majority of sexually abusive youth were male, with a family history of neglect, physical, and/or sexual abuse. Behavioral and psychological problems were common.


[Palliative endoscopic treatment of malignant occlusive jaundice with a balloon expandable metal stent].
Balloon-expandable Strecker-stents were endoscopically inserted in 13 patients (8 females, 5 men; mean age 75.2 [55-99] years) suffering from malignant obstructive jaundice. The stent implantation was difficult in one patient because of trouble to retract the balloon carrying the stent. A prompt regression of the cholestasis resulted in 11 patients. Two patients with long-distance stenoses of the common hepatic duct showed a persistent icterus due to an incomplete drainage of the stenoses. Therefore a second stent was implanted by means of transhepatic procedure in one case. Additional early complications such as cholangitis did not occur within four weeks after insertion. During the follow-up of four up to 41 weeks (mean follow-up 15.2 weeks) we observed late complications in five patients: stent migration in one patient after 10 weeks; relapsed jaundice in four patients 11 up to 41 weeks after stent implantation (the reason of jaundice was unknown for these patients were not again sent to our hospital because of their bad conditions). Six patients were alive without evidence of icterus on the date of examination. Considering data from the literature Strecker-stent implantations have been rarely performed in biliary duct stenosis till now. But the results demonstrate that an effective drainage of bile duct stenoses can be achieved in a high percentage by means of balloon-expandable metal stents. The insertion of the stent can be rather easily performed in stenoses situated in Vater's papilla and common bile duct provided that the guide wire has passed the stenosis.


The effect of beta-sitosterol on the metabolism of cholesterol and lipids in rats on a diet containing coconut oil.
1. Intraperitoneal injection of beta-sitosterol (5mg./rat/day for 25 days) into 1-year-old male Wistar rats fed on a low-fat diet supplemented with 10% of coconut oil resulted in a lowering of cholesterol and lipid concentrations in the tissues. 2. beta-Sitosterol increased the rate of biosynthesis of cholesterol and lipids in the tissues, but to an even greater extent enhanced their oxidative degradation. 3. The present results are similar to those previously obtained on a low-fat diet, indicating that the presence of fat had no marked effect on the action of beta-sitosterol.


Discs large in the Drosophila testis: an old player on a new task.
Gamete development requires a coordinated soma-germ line interaction that ensures renewal and differentiation of germline and somatic stem cells. The physical contact between the germline and somatic cell populations is crucial because it allows the exchange of diffusible signals among them. The tumor suppressor gene discs large (dlg) encodes a septate junction protein with functions in epithelial cell polarity, asymmetric neuroblast division and formation of neuromuscular junctions. Our recent work reveals a new role of dlg in the Drosophila testis, as mutations in dlg lead to testis defects and cell death. Dlg is required throughout spermatogenesis in the somatic lineage and its localization changes from a uniform distribution along the plasma membrane of somatic cells in the testis apex, to a restricted localization on the distally located somatic cell in growing cysts. The extensive defects in dlg testis underline the importance of the somatic cells in the establishment and maintenance of the male stem cell niche and somatic cell differentiation. Here, we discuss our latest findings on the role of dlg in the Drosophila testis, supporting the view that junction proteins are dynamic structures, which can provide guiding cues to recruit scaffold proteins or other signaling molecules.


The K+-translocating Kdp-ATPase from Escherichia coli. Purification, enzymatic properties and production of complex- and subunit-specific antisera.
The Kdp system from Escherichia coli is a derepressible high-affinity K+-uptake ATPase. Its membrane-bound ATPase activity was approximately 50 mumol g-1 min-1. The Kdp-ATPase complex was purified from everted vesicles by solubilization with the nonionic detergent Aminoxid WS 35 followed by DEAE-Sepharose CL-6B chromatography at pH 7.5 and pH 6.4 and gel filtration on Fractogel TSK HW-65. The overall yield of activity was 6.5% and the purity at least 90%. The isolated KdpABC complex had a high affinity for its substrates K+ (Km app. = 10 microM) and Mg2+-ATP (Km = 80 microM) and a narrow substrate specificity. The ATPase activity was inhibited by vanadate (Ki = 1.5 microM), fluorescein isothiocyanate (Ki = 3.5 microM), N,N'-dicyclohexylcarbodiimide (Ki = 60 microM) and N-ethylmaleimide (Ki = 0.1 mM). The purification protocol was likewise applicable to the isolation of a KdpA mutant ATPase which in contrast to the wild-type enzyme exhibited an increased Km value for K+ of 6 mM and a 10-fold lowered sensitivity for vanadate. Starting from the purified Kdp complex the single subunits were obtained by gel filtration on Bio-Gel P-100 in the presence of SDS. Both the native Kdp-ATPase and the SDS-denatured polypeptides were used to raise polyclonal antibodies. The specificity of the antisera was established by immunoblot analysis. In functional inhibition studies the anti-KdpABC and anti-KdpB sera impaired ATPase activity in the membrane-bound as well as in the purified state of the enzyme. In contrast, the anti-KdpC serum did not inhibit enzyme activity.


Spatial pattern formation in an insect host-parasitoid system
Spatial models in ecology predict that populations may form patchy distributions within continuous habitats, through strong predator-prey or host-parasitoid interactions combined with limited dispersal. Empirical support of these models is provided. Parasitoids emanating from a population outbreak of tussock moths (Orgyia vetusta) suppressed the growth of nearby experimental populations of the moth, while experimental populations farther away were able to grow. This result explains the observed localized nature of tussock moth outbreaks and illustrates how population distributions can be regulated by dynamic spatial processes.


Condensin I and II Complexes License Full Estrogen Receptor α-Dependent Enhancer Activation.
Enhancers instruct spatio-temporally specific gene expression in a manner tightly linked to higher-order chromatin architecture. Critical chromatin architectural regulators condensin I and condensin II play non-redundant roles controlling mitotic chromosomes. But the chromosomal locations of condensins and their functional roles in interphase are poorly understood. Here we report that both condensin complexes exhibit an unexpected, dramatic estrogen-induced recruitment to estrogen receptor α (ER-α)-bound eRNA(+) active enhancers in interphase breast cancer cells, exhibiting non-canonical interaction with ER-α via its DNA-binding domain (DBD). Condensins positively regulate ligand-dependent enhancer activation at least in part by recruiting an E3 ubiquitin ligase, HECTD1, to modulate the binding of enhancer-associated coactivators/corepressors, including p300 and RIP140, permitting full eRNA transcription, formation of enhancer:promoter looping, and the resultant coding gene activation. Collectively, our results reveal an important, unanticipated transcriptional role of interphase condensins in modulating estrogen-regulated enhancer activation and coding gene transcriptional program.


Disaster preparedness of poison control centers in the United States.
To assess the state of disaster readiness of poison control centers, a survey questionnaire was sent to all 96 institutional poison control center members of the American Association of Poison Control Centers in the US, both certified and noncertified programs. The data reported are the results and responses from 76 of 96 (79.2%) poison control centers. Fifty-four percent of responding centers have written disaster plans, with 25% having drills to practice the plans. Of the centers that do not have a written plan, the majority have policies and procedures in place to address physical plant damage, increased phone traffic, loss of phone systems and malfunction of computers. Eighty-six percent of respondents have a back up generator, and 82% have an uninterruptable power supply in place. Fifty-four percent have a back up phone system and 33% have cellular phone capacity. Forty-six percent of responding centers have arrangements with other agencies in the event of a disaster. Only half of the managing directors of the responding centers believe their center can meet the public's needs in the event of a disaster.


Use of coronary Palmaz-Schatz stent in the percutaneous treatment of vertebral artery stenoses.
A case of chronic vertebrobasilar insufficiency due to severe stenoses at the origins of both vertebral arteries was treated with percutaneous transluminal angioplasty and coronary Palmaz-Schatz stents. Use of stents led to a better angiographic result than angioplasty alone. The patient is improved 8 mo later.


Outcome of clinical and subclinical myocardial injury in systemic lupus erythematosus - A prospective cohort study.
To determine the outcome of subclinical lupus myocarditis (LM) over twelve months with regards to: mortality; incidence of clinical LM and change in imaging parameters (echocardiography and cardiac magnetic resonance [CMR]). To evaluate the impact of immunosuppression on CMR evidence of myocardial tissue injury. SLE patients with and without CMR evidence of myocardial injury (as per 2009 Lake Louise criteria [LLC]) were included. Analysis at baseline and follow-up included: clinical evaluation, laboratory and imaging analyses (echocardiography and CMR). Clinical LM was defined as clinical features of LM supported by echocardiographic and/or biochemical evidence of myocardial dysfunction. Subclinical LM was defined as CMR myocardial injury without clinical LM. Forty-nine SLE patients were included with follow-up analyses (after 12 months) available in 36 patients. Twenty-five patients (51%) received intensified immunosuppressive therapy during follow-up for indications related to SLE. Disease activity (SLEDAI-2K) improved (p < 0.001) from 13 (median;IQR:9-20) to 7 (3-11). One patient without initial CMR evidence of myocardial injury developed clinical LM. Mortality (n = 10) and SLE clinical features were similar between patients with and without initial CMR myocardial injury. Echocardiographic left ventricular ejection fraction (LVEF) (p = 0.014), right ventricular function (p = 0.001) and wall motion abnormalities (p = 0.056) improved significantly but not strain analyses nor the left LV internal diameter index. CMR mass index (p = 0.011) and LVEF (p < 0.001) improved with follow-up but not parameters identifying myocardial tissue injury (LLC). A trend towards a reduction in the presence of CMR criteria was counterbalanced by persistence (n = 7) /development of new criteria (n = 11) in patients. Change in CMR mass index correlated with change in T2-weighted signal (myocardial oedema) (r = 386;p = 0.024). Intensified immunosuppressive therapy had no significant effect on CMR parameters. CMR evidence of subclinical LM persisted despite improved SLEDAI-2K, serological markers, cardiac function and CMR mass index. Subclinical LM did not progress to clinical LM and had no significant prognostic implications over 12 months. Immunosuppressive therapy did not have any significant effect on the presence of CMR evidence of myocardial tissue injury. Improvement in CMR mass index correlated with reduction in myocardial oedema and may be used to monitor SLE myocardial injury.


The hRPC62 subunit of human RNA polymerase III displays helicase activity.
In Eukaryotes, tRNAs, 5S RNA and U6 RNA are transcribed by RNA polymerase (Pol) III. Human Pol III is composed of 17 subunits. Three specific Pol III subunits form a stable ternary subcomplex (RPC62-RPC39-RPC32α/β) being involved in pre-initiation complex formation. No paralogues for subunits of this subcomplex subunits have been found in Pols I or II, but hRPC62 was shown to be structurally related to the general Pol II transcription factor hTFIIEα. Here we show that these structural homologies extend to functional similarities. hRPC62 as well as hTFIIEα possess intrinsic ATP-dependent 3'-5' DNA unwinding activity. The ATPase activities of both proteins are stimulated by single-stranded DNA. Moreover, the eWH domain of hTFIIEα can replace the first eWH (eWH1) domain of hRPC62 in ATPase and DNA unwinding assays. Our results identify intrinsic enzymatic activities in hRPC62 and hTFIIEα.


A new method of measuring the rate of shedding of epithelial cells from the intestinal villus of the rat.
Epithelial cells shed from the intestinal villus in starved rats were trapped in mucus which the goblet cells were stimulated to secrete; the number of trapped cells per villus was counted in whole mounts of fixed Feulgen-stained specimens from three sites in the small intestine. Increase in the interval between stimulation and fixation caused an increase in the number of shed cells per villus; the rate of this increase was postulated to represent the rate of cell shedding. The rate of cell production per crypt was measured with Colcemid; this rate, multiplied by the number of crypts per villus, gave the rate of cell production per villus. There was good agreement between these estimates of cell production and cell shedding.


Comparative Analysis of Postoperative Complications after Cytoreductive Surgery and HIPEC in Gastric Cancer.
Patients with advanced gastric cancer (AGC) frequently show peritoneal carcinomatosis (PC). PC reduces life expectancy and quality of life. Cytoreductive surgery (CRS) with hyperthermic intraperitoneal chemotherapy (HIPEC) has been shown to improve overall survival. Nevertheless, it has been reported that CRS and HIPEC are accompanied by an increase in postoperative complications. The purpose of this study was to investigate the complications associated with CRS and HIPEC and overall and disease-free survival. Patients with AGC and PC, who received complete CRS and HIPEC, were included in the HIPEC group (n = 15). Patients with AGC but without PC, who received resection of the primary tumor alone, constituted the control group (n = 43). Patients enrolled in the HIPEC group presented with a median PCI of 7. In comparison with the control group, no differences were found in patient characteristics, risk factors, pathological findings, and operative procedures. Twenty-five percentage of the patients in both groups suffered from serious postoperative complications (CDC ≥3a). Surgical and medical complications, rate of reoperation, and mortality did not differ. Also, the recurrence pattern, median survival, and 1- and 2-year survival rates showed no differences. CRS and HIPEC do not lead to an increased postoperative morbidity and mortality in AGC with PC. Albeit the poorer prognosis of patients with PC, survival of both groups was comparable.


Alterations of neuroendocrine brain asymmetry: a neural risk factor affecting physical health.
It was examined whether the specific pattern of brain asymmetry in cerebral control of cortisol secretion existing in a specific individual is suitable to predict the individual's physical health status. Results indicate that subjects with an altered, strong left hemisphere dominance of cortisol regulation show a clearly higher amount of physical complaints than subjects with a normal, strong right hemisphere dominance. In addition, results suggest that the subjects' specific pattern of neuroendocrine brain asymmetry can be used with relatively good success to separate individuals with high versus low degrees of physical complaints and thus, probably, to predict physical complaints of an individual.


High-speed tracking control of piezoelectric actuators using an ellipse-based hysteresis model.
In this paper, an ellipse-based mathematic model is developed to characterize the rate-dependent hysteresis in piezoelectric actuators. Based on the proposed model, an expanded input space is constructed to describe the multivalued hysteresis function H[u](t) by a multiple input single output (MISO) mapping Gamma:R(2)-->R. Subsequently, the inverse MISO mapping Gamma(-1)(H[u](t),H[u](t);u(t)) is proposed for real-time hysteresis compensation. In controller design, a hybrid control strategy combining a model-based feedforward controller and a proportional integral differential (PID) feedback loop is used for high-accuracy and high-speed tracking control of piezoelectric actuators. The real-time feedforward controller is developed to cancel the rate-dependent hysteresis based on the inverse hysteresis model, while the PID controller is used to compensate for the creep, modeling errors, and parameter uncertainties. Finally, experiments with and without hysteresis compensation are conducted and the experimental results are compared. The experimental results show that the hysteresis compensation in the feedforward path can reduce the hysteresis-caused error by up to 88% and the tracking performance of the hybrid controller is greatly improved in high-speed tracking control applications, e.g., the root-mean-square tracking error is reduced to only 0.34% of the displacement range under the input frequency of 100 Hz.


Stability of extracts from pollens of allergenic importance in Korea.
Accurate diagnosis and the effects of allergen-specific immunotherapy for pollinosis are greatly dependent on the potency and stability of the extract. This study aimed to examine factors, such as temperature and storage buffer composition, that affect the stability of allergen extracts from pollens of allergenic importance in Korea. We prepared four pollen allergen extracts from ragweed, mugwort, Japanese hop, and sawtooth oak, which are the most important causes of seasonal rhinitis in Korea. Changes of protein and major allergen concentration were measured over 1 year by Bradford assay, two-site enzyme-linked immunosorbent assay, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis after reconstitution of the lyophilized allergen extract in various buffers and stored at room temperature (RT, 18°C to 26°C) or refrigerated (4°C). More than 90% of the original protein concentration in all four extracts examined was detected over 1 year when 50% glycerol was added and refrigerated, whereas 57.9% to 94.5% remained in the extracts at RT. The addition of 50% glycerol to the storage buffer was found to prevent protein degradation at RT. Amb a 1, a major allergen of ragweed, was almost completely degraded in 9 weeks at RT when reconstituted in a buffer without 50% glycerol. However, 55.6% to 92.8% of Amb a 1 content was detected after 1 year of incubation at 4°C in all buffer conditions except 0.3% phenol. Addition of 50% glycerol as well as refrigeration was found to be important in increasing the shelf-life of allergen extracts from pollens of allergenic importance.


[Effects of dissolved compounds on photodegradation of mefenacet in water].
The study with high-pressure mercury lamp illuminating showed that after illuminated for 15 min, NO2- and NO3- quenched the photolysis of mefenacet, and NO3- with a concentration ratio 10:1 (mass) had the most obvious effect, its quenching rate being up to 53.3%. Halogen ions inhibited the photolysis of mefenacet by "weight atom effect". When the concentration ratio of I- was 10:1, the quenching rate was 76.9% after illuminated for 15 min. Surfactants Nongru 500, Nongru 404, Nongru 601 and Nongru 603 had different effects on the photodegradation of mefenacet. At concentration ratios 1:5 and 1:1, only Nongru 404 showed a weak photosensitive effect, while in the other cases, all the four surfactants had photoquenching effects. Among the four herbicides benthiocarb, bensulfuron, alachlor and chlorsulfuron, only bensulfuron at low concentration ratio (1 : 10) accelerated the photolysis of mefenacet, with a photosensitive proportion of 18.2% after illuminated for 25 min. Aerified N2 could accelerate the photolysis of mefenacet, and the half-life was shortened from 7.14 min to 6.70 min without aerifying N2.


STAT3 modulates the DNA damage response pathway.
The STAT3 transcription factor is well known to function as an anti-apoptotic factor, especially in numerous malignancies. Recently we showed that STAT3 is cytoprotective and that cells lacking STAT3 are more sensitive to oxidative stress. A key feature of oxidative stress involves activation of the DNA damage pathway. However, a role for STAT3 or its contribution in response to DNA damage has not been described. In the present study we show that cells lacking STAT3 are less efficient in repairing damaged DNA. Moreover, STAT3 deficient cells show reduced activity of the ATM-Chk2 and ATR-Chk1 pathways, both important pathways in sensing DNA damage. Finally we show that MDC1, a regulator of the ATM-Chk2 pathway and facilitator of the DNA damage response, is modulated by STAT3 at the transcriptional level. These findings demonstrate that STAT3 is necessary for efficient repair of damaged DNA, partly by modulating the ATM-Chk2 and ATR-Chk1 pathways.


Current use and perceived utility of ultrasound for evaluation of pediatric compared with adult trauma patients.
The objective of this study was to evaluate the current use and perceived utility of ultrasound in the assessment of pediatric compared with adult trauma patients. A questionnaire was developed and mailed to 72 pediatric emergency physicians, 120 general emergency physicians, and 117 trauma attendings at 240 institutions. Of 309 surveys, 234 (75%) were completed. Ultrasound was available to 169 of 234 (72%) of the physicians, and 122 of 169 (72%) were performing the Focused Assessment by Sonography for Trauma examination to evaluate trauma patients. Seventy-three percent (110/150) of general emergency and trauma surgeons reported that ultrasound was available equally with or more readily than computed tomography (CT) scan. Only 26% (5/19) of pediatric emergency attendings considered ultrasound equally available with CT scan, and none considered it more readily available than CT scan. Ninety-two percent (137/149) of general emergency and trauma attendings responding to the question about utility considered ultrasound somewhat useful to extremely useful for assessing adult trauma patients, and 77% considered it useful for pediatric patients. Only 57% (12/21) of pediatric emergency attendings responding to the same question perceived ultrasound as useful for pediatric trauma evaluations. We conclude that ultrasound for the assessment of trauma patients is widely used by general emergency physicians and trauma surgeons, whereas pediatric emergency physicians report less use and perceived utility.


Therapeutic drug monitoring for antidepressants: efficacy, safety, and cost effectiveness.
Therapeutic drug monitoring (TDM) of tricyclic antidepressants (TCAs) is a cost-effective tool to increase efficacy and safety in patients suffering from major depressive illness. The concentration:antidepressant response relationships of nortriptyline, desipramine, and amitriptyline conform to a curvilinear relationship between concentration:antidepressant response; whereas, imipramine conforms to a linear relationship. Besides improving efficacy, TDM identifies the subset of patients who are at risk to develop excessive plasma TCA levels with potential resultant central nervous system and cardiovascular toxicity. Taking both the efficacy and safety data into account, cost:benefit analyses demonstrate that TDM is cost effective when used once as a standard aspect of care in the TCA treatment of patients with major depressive illness.


Association Between Genetic Polymorphisms and Pain Sensitivity in Patients with Hip Osteoarthritis.
Factors such as age, gender, and genetic polymorphisms may explain individual differences in pain phenotype. Genetic associations with pain sensitivity have previously been investigated in osteoarthritis patients, with a focus on the P2X7, TRPV1, and TACR1 genes. However, other genes may play a role as well. Osteoarthritis is a common joint disease, and many patients suffering from this disease are thought to have increased sensitivity to noxious stimuli resulting from sensitization in the nociceptive system. The aim of this study was to investigate if genetic variants of mu, kappa, and delta opioid receptor genes (OPRM1, OPRK1, and OPRD1) and the catechol-O-methyltransferase gene (COMT) influenced the pain phenotype in patients with osteoarthritis. The frequencies of 17 polymorphisms were examined. Pain sensitivity was assessed preoperatively by (1) hip rotation, (2) contact heat stimulation, (3) conditioned pain modulation effect, and (4) pressure stimulation at the tibia in both the affected and the unaffected leg. Ninety-two patients (mean age 66 years) with unilateral hip osteoarthritis were included in the study. Carriage of the OPRM1 rs589046T allele was found to be associated with increased pain ratings during hip rotation (P = 0.04) and increased conditioned pain modulation (P = 0.049). Carriage of the OPRD1 rs2234918C allele was found to be associated with an increased pain detection threshold to contact heat stimulation (P = 0.001). No other associations were found (all P > 0.05). Results from the present study suggest that, in patients with hip osteoarthritis, genetic variants in OPRM1 and OPRD1 may contribute to the pain phenotype.


Impact of Introduction of a Clinical Pathway for the Management of Pyelonephritis on Obstetric Patients: a Quality Improvement Project
Aim To determine whether the introduction of a clinical pathway for the treatment of pyelonephritis in obstetric patients would improve outcomes. Methods This was a retrospective study conducted in a maternity hospital using quantitative analysis methods. Patients who met laboratory and clinical criteria for pyelonephritis during data collection were included. Results The study included analysis of 23 patients pre-intervention and 19 post-intervention. Baseline and patient characteristics were similar for both groups. A statistically significant difference was seen in 3 of 7 outcome measures. Increased use of gentamicin (13% Vs 52% p=0.006), Increased number of renal ultrasounds (17% Vs 47%, p=0.04) and increased use of prophylaxis (21% Vs 68%, p=0.003). The proportion of patients receiving ≥ 10 days of IV antimicrobials decreased from 48% to 21% post-intervention (p=0.07). Discussion This study has shown that the introduction of a pathway for the treatment of pyelonephritis in pregnancy had a positive impact on several important clinical outcomes.


Estimation of pollutant loads from urban roadway runoff.
A simulation model consisting of a deposition process and a wash-off process was proposed to evaluate the pollutant loads from urban roadways, and was verified based on field survey data obtained over a 5-year period in the Lake Biwa watershed. The model parameters were determined by minimizing the total sum of squares of differences between the observed data and simulated ones. By applying this model to all roadways in the watershed, the calculated amounts of COD(Mn), organic carbon, nitrogen and phosphorus in particulate forms were 15.9, 15.7, 0.88 and 0.15 kg/(km(2) .d), respectively, and those in dissolved forms were 14.1, 12.5, 2.62 and 0.03 kg/(km(2) .d), respectively. From the results, the pollutant loads of COD(Mn), TN and TP obtained for the Lake Biwa watershed (total roadway area of 98.9 km(2)) were estimated to be 2,950, 350 and 18 kg/d, respectively.


Dynamics of a Polymer Network Based on Dual Sierpinski Gasket and Dendrimer: A Theoretical Approach.
In this paper we focus on the relaxation dynamics of a multihierarchical polymer network built through the replication of the dual Sierpinski gasket in the form of a regular dendrimer. The relaxation dynamics of this multihierarchical structure is investigated in the framework of the generalized Gaussian structure model using both Rouse and Zimm approaches. In the Rouse-type approach, we show a method whereby the whole eigenvalue spectrum of the connectivity matrix of the multihierarchical structure can be determined iteratively, thereby rendering possible the analysis of the Rouse-dynamics at very large generations. Remarkably, the general picture that emerges from both approaches, even though we have a mixed growth algorithm and the monomers interactions are taken into account specifically to the adopted approach, is that the multihierarchical structure preserves the individual relaxation behaviors of its constituent components. The theoretical findings with respect to the splitting of the intermediate domain of the relaxation quantities are well supported by experimental results.


Women with mechanical neck pain exhibit increased activation of their superficial neck extensors when performing the cranio-cervical flexion test.
Several studies have analysed the activity of superficial neck flexors, but the activity of neck extensors has been less investigated in patients with neck pain OBJECTIVES: 1, to investigate the differences in the activation of superficial neck flexor and extensor musculature during the cranio-cervical flexion test (CCFT) in women with mechanical chronic neck pain when compared to asymptomatic women; 2, to investigate the correlation between neck muscle activity and the clinical features of neck pain DESIGN: Cross-sectional METHODS: Surface electromyography was recorded bilaterally from the sternocleidomastoid, anterior scalene, splenius capitis, and upper trapezius muscles of 30 women with mechanical chronic neck pain and 30 asymptomatic women as they performed the CCFT. Comparisons of the normalized root mean square between both groups were conducted with 2x5 ANCOVA with task level as the within-subjects variable, group as the between-subjects variable, and pain related-disability as a co-variate RESULTS: Women with mechanical neck pain exhibited increased activity of superficial neck flexors (sternocleidomastoid: F = 14.448, P < 0.001; anterior scalene: F = 21.693, P < 0.001) and superficial neck extensors (splenius capitis: F = 4.692, P < 0.001; upper trapezius: F = 4.245, P < 0.001) as compared to asymptomatic women. Higher pain related-disability was associated with more electrical activity of the anterior scalene and upper trapezius muscles during the CCFT CONCLUSIONS: Women with mechanical chronic neck pain exhibit an increased activity of their superficial neck flexors and superficial neck extensors during a low-load task such as CCFT when compared to asymptomatic pain-free women. Our results should be considered when designing therapeutic exercise programs for this population.


Global hypermethylation of the N6-methyladenosine RNA modification associated with apple heterografting.
Grafting can facilitate better scion performance and is widely used in plants. Numerous studies have studied the involvement of mRNAs, small RNAs, and epigenetic regulations in the grafting process. However, it remains unclear whether the mRNA N6-methyladenosine (m6A) modification participates in the apple (Malus x domestica Borkh.) grafting process. Here, we decoded the landscape of m6A modification profiles in 'Golden delicious' (a cultivar, Gd) and Malus prunifolia 'Fupingqiuzi' (a unique rootstock with resistance to environmental stresses, Mp), as well as their heterografted and self-grafted plants. Interestingly, global hypermethylation of m6A occurred in both heterografted scion and rootstock compared with their self-grafting controls. Gene Ontology (GO) term enrichment analysis showed that grafting-induced differentially m6A-modified genes were mainly involved in RNA processing, epigenetic regulation, stress response, and development. Differentially m6A-modified genes harboring expression alterations were mainly involved in various stress responses and fatty acid metabolism. Furthermore, grafting-induced mobile mRNAs with m6A and gene expression alterations mainly participated in ABA synthesis and transport (e.g. carotenoid cleavage dioxygenase 1 [CCD1] and ATP-binding cassette G22 [ABCG22]) and abiotic and biotic stress responses, which might contribute to the better performance of heterografted plants. Additionally, the DNA methylome analysis also demonstrated the DNA methylation alterations during grafting. Downregulated expression of m6A methyltransferase gene MdMTA (ortholog of METTL3) in apples induced the global m6A hypomethylation and distinctly activated the expression level of DNA demethylase gene MdROS1 (REPRESSOR OF SILENCING 1) showing the possible association between m6A and 5mC methylation in apples. Our results reveal the m6A modification profiles in the apple grafting process and enhance our understanding of the m6A regulatory mechanism in plant biological processes.


Leveraging an information warehouse to create translational research environment for wound care center.
In order to discover new biomarkers and therapeutic agents for personalized wound care, a vast amount of clinical information is collected and stored at The Ohio State University Medical Center (OSUMC) Comprehensive Wound Center (CWC). The Information Warehouse (IW) group at OSUMC has developed and implemented a comprehensive data collection network and analysis pipeline to support clinical, translational and outcomes research, and cost analyses that can be converted into clinical best practices for wound care.


pRb controls estrogen receptor alpha protein stability and activity.
A cross talk between the Estrogen Receptor (ESR1) and the Retinoblastoma (pRb) pathway has been demonstrated to influence the therapeutic response of breast cancer patients but the full mechanism remains poorly understood. Here we show that the N-terminal domain of pRb interacts with the CD domain of ESR1 to allow for the assembly of intermediate complex chaperone proteins HSP90 and p23. We demonstrated that a loss of pRb in human/mouse breast cells decreases the expression of the ESR1 protein through the proteasome pathway. Our work reveals a novel regulatory mechanism of ESR1 basal turnover and activity and an unanticipated relationship with the pRb tumor suppressor.


Suspected COVID-19-related reticulated purpura of the soles in an infant.
A growing number of skin lesions during the COVID-19 pandemic are being recognized. Acral ischemic lesions identical to chilblains are most typical in children and young adults. We report an infant girl, aged 1 month and 29 days, with a peculiar reticulated purpuric eruption on her soles, with positive immunohistochemistry for SARS-CoV-2 in the endothelia of dermal blood vessels. The patient had an excellent outcome without specific therapy.


Solution structure of heavy meromyosin by small-angle scattering.
Elucidation of x-ray crystal structures for the S1 subfragment of myosin afforded atomic resolution of the nucleotide and actin binding sites of the enzyme. The structures have led to more detailed hypotheses regarding the mechanisms by which force generation is coupled to ATP hydrolysis. However, the three-dimensional structure of double-headed myosin consisting of two S1 subfragments has not yet been solved. Therefore, to investigate the overall shape and relative orientations of the two heads of myosin, we performed small-angle x-ray and neutron scattering measurements of heavy meromyosin containing all three light chains (LC(1-3)) in solution. The resulting small-angle scattering intensity profiles were best fit by models of the heavy meromyosin head-tail junction in which the angular separation between heads was less than 180 degrees. The S1 heads of the best fit models are not related by an axis of symmetry, and one of the two S1 heads is bent back along the rod. These results provide new information on the structure of the head-tail junction of myosin and indicate that combining scattering measurements with high resolution structural modeling is a feasible approach for investigating myosin head-head interactions in solution.


Synergistic antiproliferative activity of tumor necrosis factor alpha (TNF-alpha) and lovastatin.
We assessed the antiproliferative effect of tumor necrosis factor alpha (TNF-alpha) and lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, alone and in combination, on two murine tumor cell lines. Recombinant TNF-alpha inhibited proliferation of murine MmB16 melanoma cells in a concentration-dependent fashion but stimulated growth of murine L1210 leukemia cells at 0.1 ng/ml concentration. Lovastatin inhibited proliferation both of murine MmB16 melanoma cells and of murine L1210 leukemia cells in a concentration-dependent fashion. In combination with tumor necrosis factor alpha lovastatin inhibited synergistically growth of both cell lines as assessed by isobologram analysis. Our data show that lovastatin, a cholesterol synthesis inhibitor, introduced to the clinic to treat hypercholesterolemia, used either as a single or in combination with TNF-alpha inhibits growth of MmB16 melanoma and L1210 leukemia cells.


A rare association of left pulmonary artery sling with right pulmonary hypoplasia and total anomalous pulmonary venous connection.
We present the second reported case of left pulmonary artery sling with dextrocardia, right pulmonary hypoplasia, and total pulmonary venous connection in a fetus. This case highlights the importance of the determination of pulmonary artery arrangement by fetal echocardiography if right pulmonary hypoplasia and/or congenital heart disease is suspected.


Pannexin1 in the outer retina of the zebrafish, Danio rerio.
In the retina, chemical and electrical synapses couple neurons into functional networks. New candidates encoding for electrical synapse proteins have recently emerged. In the present study, we determined the localization of the candidate protein pannexin1 (zfPanx1) in the zebrafish retina and studied the functional properties of zfPanx1 exogenously expressed in Neuroblastoma 2a (N2a) cells. zfPanx1 was identified on the surface of horizontal cell dendrites invaginating deeply into the cone pedicle near the glutamate release sites of the cones, providing in vivo evidence for hemichannel formation at that location. This strategic position of zfPanx1 in the photoreceptor synapse could potentially allow modulation of cone output. Using whole cell voltage clamp and excised patch recordings of transfected N2a cells, we demonstrated that zfPanx1 forms voltage-activated hemichannels with a large unitary conductance in vitro. These channels can open at physiological membrane potentials. Functional channels were not formed following mutation of a single amino acid within a conserved protein motif recently shown to be N-glycosylated in rodent Panx1. Together, these findings indicate that zfPanx1 displays properties similar to its mammalian homologues and can potentially play an important role in functions of the outer retina.


Strong Association of Angiotensin Converting Enzyme-2 Gene Insertion/Deletion Polymorphism with Susceptibility to SARS-CoV-2, Hypertension, Coronary Artery Disease and COVID-19 Disease Mortality.
The ongoing outbreak of SARS-CoV-2 represents a significant challenge to international health. Several reports have highlighted the importance of ACE2 on the pathogenesis of COVID-19. The spike protein of SARS-CoV-2 efficiently binds to the angiotensin-converting enzyme 2 (ACE2) receptors and facilitates virus entry into the host cell. In the present study, we hypothesize that a functional insertion/deletion polymorphism-rs4646994 I/D and rs4240157 T > C in the ACE gene could be associated with SARS-CoV-2 infection and mortality. This study included 117 consecutive COVID-19 patients and 150 age matched healthy controls (ACE2-rs4646994 I/D) and 100 age matched healthy controls with ACE2 rs4240157 T > C. We used Mutation specific PCR (MSP) for ACE2-rs4646994 I/D genotyping and amplification refractory mutation system (ARMS-PCR) for ACE2 rs4240157 T > C genotyping. Results indicated that there were significant differences in the genotype distributions of ACE2-rs4646994 I/D polymorphisms (p < 0.030) and ACE2 rs4240157 T > C between COVID-19 patients and controls (p-values < 0.05). Higher frequency of DD genotype (48.71%) and D allele (0.67) was reported in COVID-19 patients than controls. Our results showed that the ACE2-DD genotype was strongly associated with increased COVID-19 severity (OR 2.37 (95%) CI = (1.19-4.70), RR = 1.39 (1.09-1.77), p < 0.013) and also a strong association was seen with ACE2-ID genotype with COVID-19 severity (OR 2.20 (95%) CI = (1.08-4.46), p < 0.020) in the codominant model. In allelic comparison, the D allele was strongly associated with COVID-19 severity (OR 1.58 (95% CI) (1.11-2.27), RR 1.21 (1.05-1.41) p < 0.010). A significant correlation of ACE2-I/D genotypes was reported with Age (p < 0.035), T2D (p < 0.0013), hypertension (p < 0.0031) and coronary artery disease (p < 0.0001). Our results indicated ACE2-DD genotype was strongly associated with increased COVID-19 mortality (OR 8.25 (95%) CI = (2.40 to 28.34), p < 0.008) and also ACE2-DD + DI genotype was strongly associated with increased COVID-19 mortality with OR 4.74 (95%) CI = (1.5214 to 14.7915), p < 0.007. A significant correlation was reported between COVID-19 patients and age matched controls (p < 0.0007). Higher frequency of heterozygosity TC (40%) followed by ACE2-CC genotype (24.78%) was reported among COVID-19 patients. Using multivariate analysis, ACE2-CT genotype was strong associated with SARS-CoV-2 severity with an OR 2.18 (95% CI) (1.92-3.99), p < 0.010 and also ACE2-CC genotype was linked with COVID-19 severity with an OR 2.66 (95% CI) (1.53-4.62), p < 0.005. A significant correlation of ACE2-T > C genotypes was reported with gender (p < 0.04), T2D (p < 0.035). ACE2-CC genotype was strongly associated with increased COVID-19 mortality OR 3.66 (95%) CI = (1.34 to 9.97), p < 0.011 and also ACE2-C allele was associated with COVID-19 mortality OR 2, 01 (1.1761-3.45), p < 0.010. It is concluded that ACE-DD genotype and D allele was strongly associated with increased COVID-19 patient severity. In addition, ACE I/D polymorphism were strongly associated with advanced age, diabetes and ischemic heart disease in COVID-19 patients whereas ACE-II genotype was a protective factor against the development of severe COVID-19. ACE2-DD genotype was strongly associated with increased COVID-19 mortality. Additionally, ACE2-CC and CT genotypes were strongly associated with COVID-19 severity. Therefore, our study might be useful for identifying the susceptible population groups for targeted interventions and for making relevant public health policy decisions.


Structure of the fifth EGF-like domain of thrombomodulin: An EGF-like domain with a novel disulfide-bonding pattern.
The structure of the fifth EGF-like domain (residues Q387 to E426) of thrombomodulin (TMEGF5) has been determined by two-dimensional NMR. TMEGF5 binds to thrombin with a Ki of 1.9 microM and has been shown to have a novel disulfide bonding pattern in a fully active fragment of TM. In EGF, the disulfide bonding pattern is (1-3,2-4, 5-6), while TMEGF5 has an uncrossed (1-2,3-4,5-6) pattern. The structure of this novel domain, determined from 483 NOE-derived distance restraints, appears to have diverged from the common EGF-like structure. Superposition of the 14 lowest-energy structures of TMEGF5 gives an overall r.m.s.d. of 1.09 A for the backbone atoms. The central two-stranded beta-sheet common to all EGF-like domains is not present in TMEGF5. The A loop, residues C390 to C395, is twisted away from interacting with the B loop, residues C399 to C407, as in EGF, and is close to the C loop, residues C409 to C421. This twist causes the N and C termini to be closer together in TMEGF5 than in EGF. Most of the residues that are important for activity lie on one face of the molecule, which is likely to be the thrombin-binding surface of the domain. The structure of the C loop within the domain, which is a beta-hairpin similar to EGF, is similar to the structure of a synthetic version of the loop bound to thrombin as determined by transferred NOE experiments. Despite the similarity in the structures of the loops, the residues immediately following C421 are in different positions in the two structures suggesting that these "tail" residues may change conformation upon thrombin binding.


[Diagnosis and treatment of primary thrombocythemia].
Primary thrombocythemia is a frequent myeloproliferative disorder that may be asymptomatic for a long period of time. Once the usual reactive causes have been excluded, further diagnosis requires specialized exploration. The severity level of the disease is not necessarily related to reduce survival rates, but to thrombotic or hemorrhagic complications that can affect quality of life. Myelofibrotic and acute leukemic transformations can also be long-term complications, possibly influenced by treatment strategy. Cytoreductive treatment is required in patients older than 60, or with a history of thrombosis or hemorrhage, or with a platelet count above 1 500 - 109/L. In cases where these indicators do not exist, there are no guidelines for treatment in France, and all investigators try to discriminate patients with a low vascular risk in order to avoid cytoreductive treatment. In fact, in the absence of curative treatment, it seems reasonable to avoid potentially leukemogenic drugs in younger patients.


Prediction of Prostate Cancer Aggressiveness Using a Novel Multiparametric Magnetic Resonance Imaging Parameter: Tumor Heterogeneity Index.
The aim of this study was to evaluate if the tumor heterogeneity index can predict the aggressiveness of prostate cancer (PCa) in patients diagnosed by magnetic resonance imaging (MRI) fusion biopsy. Patients who underwent MRI fusion prostatic biopsy between July 2019 and December 2020 were retrospectively reviewed. Tumor heterogeneity index (coefficient of variation [CV]) and PI-RADS v2.1 scoring were analyzed by using multiparametric MRI. The patients were divided into 3 groups according to the risk classification, and the correlation between tumor heterogeneity index and PCa aggressiveness was studied by using apparent diffusion coefficient (ADCmean and ADCcv), Gleason score (GS), and risk classifications. One hundred two patients were included in this study. Patients were evaluated as low-risk (group 1) (n = 35), moderate-risk (group 2) (n = 37), and high-risk (group 3) (n = 30). ADCmean values for all groups were significantly different (p < 0.0001). ADCcv tumor heterogeneity index values were higher in group 2 and group 3 by the score increases in subgroups according to GS, while being higher than group 1 (p < 0.001). The multivariate analysis revealed that prostate-specific antigen, PI-RADS, ADCmean, and ADCcv values were predictive for tumor aggressiveness. ADCcv value as a tissue texture parameter can be used as a new biomarker to evaluate tumor aggressiveness in patients with PCa.