Shear wave elastography to evaluate carotid artery elasticity in long-term drinkers with varying degrees of alcoholic fatty liver disease.
To evaluate carotid artery elasticity in long-term drinkers with varying degrees of alcoholic fatty liver disease (AFLD) by shear wave elastography (SWE). Of the 92 drinkers with alcohol drinking for greater than or equal to 5 years, were fell into three groups depending on the liver ultrasound results: group B (without AFLD), group C (with mild AFLD), and group D (moderate-to-severe AFLD). Another 32 healthy adults were selected as the control group (group A). All participants had no significant carotid plaque. Gray scale ultrasound and color doppler ultrasound were used to obtain the left common carotid peak systolic velocity (PSV), carotid diastolic internal diameter (Dd ), carotid intima-media thickness (IMT), carotid systolic internal diameter (Ds ) and the stiffness coefficient (β). Mean values of mean elastic modulus (MEmean ), minimum elastic modulus (MEmin ), and maximum elastic modulus (MEmax ) of the anterior wall of the left common carotid artery at end-diastole were measured by SWE. Dd , Ds , and PSV among the four groups showed no differences. (all P > 0.05). Compared with groups A and B, MEmin , MEmax , and MEmean were significantly higher in group C (all P < 0.05), while IMT and β were not statistically different (all P > 0.05). MEmean , MEmin , MEmax , IMT, and β were significantly higher in group D compared with the other three groups (all P < 0.05). No significant differences were observed in IMT, β, MEmean , MEmax , and MEmin between groups A and B (all P > 0.05). Shear wave elastography can provide a quantitative evaluation for the carotid artery elasticity in long-term drinkers with varying degrees of AFLD.


Association between dyslipidemia and asthma in children: a systematic review and multicenter cohort study using a common data model.
The association between dyslipidemia and asthma in children remains unclear. This study investigated the association between dyslipidemia and cholesterol levels in children. A systematic literature review was performed to identify studies investigating the association between dyslipidemia and asthma in children. The PubMed database was searched for articles published from January 2000-March 2022. Data from a cohort study using electronic health records from 5 hospitals, converted to the Observational Medical Outcomes Partnership Common Data Model (OMOP-CDM), were used to identify the association between total cholesterol (TC) levels and asthma in children. This cohort study used the Cox proportional hazards model to examine hazard ratio (HR) of asthma after propensity score matching, and included an aggregate meta-analysis of HR. We examined 11 studies reporting an association between dyslipidemia and asthma in children. Most were cross-sectional; however, their results were inconsistent. In OMOP-CDM multicenter analysis, the high TC (>170 mg/dL) group included 29,038 children, while the normal TC (≤170 mg/dL) group included 88,823 children including all hospital datasets. In a meta-analysis of this multicenter cohort, a significant association was found between high TC levels and later development of asthma in children <15 years of age (pooled HR, 1.30; 95% confidence interval, 1.12-1.52). Elevated TC levels in children may be associated with asthma.


Reliable variables in the exsanguinated patient which indicate damage control and predict outcome.
Exsanguination as a syndrome is ill defined. The objectives of this study were to investigate the relationship between survival and patient characteristics--vital signs, factors relating to injury and treatment; determine if threshold levels of pH, temperature, and highest estimated blood loss can predict survival; and identify predictive factors for survival and to initiate damage control. A retrospective 6-year study was conducted, 1993 to 1998. In all, 548 patients met one or more criteria: (1) estimated blood loss > or =2,000 mL during trauma operation; (2) required > or =1,500 mL packed red blood cells (PRBC) during resuscitation; or (3) diagnosis of exsanguination. Analysis was made in two phases: (1) death versus survival in emergency department (ED); (2) death versus survival in operating room (OR). Statistical methods were Fisher's exact test, Student's t test, and logistic regression. For 548 patients, mean Revised Trauma Score 4.38, mean Injury Severity Score 32. Penetrating injuries 82% versus blunt injuries 18%. Vital statistics in emergency department: mean blood pressure 63 mm Hg, heart rate 78 beats per minute. Mean OR pH 7.15 and temperature 34.3 degrees C. Mortality was 379 of 548 (69%). Predictive factors for mortality (means): pH < or =7.2, temperature <34 degrees C, OR blood replacement >4,000 mL, total OR fluid replacement >10,000 mL, estimated blood loss >15 mL/minute (P <0.001). Analysis 1: death versus survival in ED, logistic regression. Independent risk factors for survival: penetrating trauma, spontaneous ventilation, and no ED thoracotomy (P <0.001; probability of survival 0.99613). Analysis 2: death versus survival in OR, logistic regression. Independent risk factors for survival: ISS < or =20, spontaneous ventilation in ED, OR PRBC replacement <4,000 mL, no ED or OR thoracotomy, absence of abdominal vascular injury (P <0.001, max R(2) 0.55, concordance 89%). Survival rates can be predicted in exsanguinating patients. "Damage control" should be performed using these criteria. Knowledge of these patterns can be valuable in treatment selection.


Sonographic myometrial thickness predicts the latency interval of women with preterm premature rupture of the membranes and oligohydramnios.
Term labor is associated with global thinning of the myometrium. We hypothesized that a thickened myometrium at the time of preterm premature rupture of membranes (PPROM) predicts less myometrial wall stress and, consequently, a longer latency interval. Myometrial thickness was measured prospectively in 76 pregnant women enrolled in the following groups: PPROM (n=28, mean [range], gestational age [GA]: 29.5 weeks [w] [21.0 w-33.0 w]), preterm nonlabor control group (P-CTR), (n=21, GA: 27.5 w [23.0 w-32.0 w]) and term nonlabor control (T-CTR) (n=27, GA: 38.6 w [37.0 w-41.6 w]). All PPROM women had oligohydramnios (AFI: 1.4 cm [0.0 cm-5.1 cm]). MT was measured ultrasonographically at the midanterior, fundal, posterior, and lower uterine segment wall in cases and controls with an intraoperator variability <10%. Women in the PPROM group displayed uniform thickness of the uterine body (mean +/- SEM, anterior: 10.6 +/- 0.6 mm, fundal: 10.7 +/- 0.7 mm, posterior: 8.9 +/- 0.5 mm, P=.078). At midanterior site the myometrium of the PPROM group was thicker compared to both P-CTR (P < .001) and T-CTR (P=.025) groups. This difference was preserved at the fundus (PPROM vs P-CTR, P < .001; PPROM vs T-CTR, P=.015). There was a positive correlation between fundal MT and latency period (r=0.43, P=0.02) that persisted after adjusting for GA (P=.04). A fundal MT less than 12.1 mm was 93.7% sensitive and 63.6% specific for the identification of women whose latency period was less than 120 hours. Significant thickening of the anterior and fundal walls of the uterus follows PPROM. A thick myometrium in nonlaboring patients with PPROM is associated with longer latency interval. Sonographic evaluation of MT may represent an alternative clinical tool for the prediction of a short latency interval in women with PPROM.


Studies on pyrazine derivatives. Part VIII. Synthesis and tuberculostatic activity of some 6-alkylthio- and 6-phenylthiopyrazine-2-carboxylic acids.
2-Cyano-6-alkylthio- and 6-phenylthiopyrazines were prepared from 2-cyano-6-chloropyrazine (CCP) in the reaction with mercaptans. By the transformation of the -CN group the corresponding amides, thioamides, amidoximes, acids, esters and hydroxamic acids and hydrazides were obtained. The oxidation of sulfur in the above mentioned compounds led to sulfoxy- and sulfonyl derivatives. In the screening for antituberculous activity compounds 30 and 42 showed the greatest activity (MIC = 31.2 microgram/cm3) against H37Rv strain and compund 8 so did against saprophytic ATCC 607 strain. The compounds with 6-benzylthio grouping were more active against ethionamide, isoniazide and capreomycine resistant strains.


Radiosensitizing effect of epothilone B on human epithelial cancer cells.
A combined modality treatment employing radiation and chemotherapy plays a central role in the management of solid tumors. In our study, we examined the cytotoxic and radiosensitive effect of the microtubule stabilizer epothilone B on two human epithelial tumor cell lines in vitro and its influence on the microtubule assembly. Cancer cells were treated with epothilone B in proliferation assays and in combination with radiation in colony-forming assays. For the analysis of ionizing radiation-induced DNA damage and the influence of the drug on its repair a γH2AX foci assay was used. To determine the effect of epothilone B on the microtubule assembly in cells and on purified tubulin, immunofluorescence staining and tubulin polymerization assay, respectively, were conducted. Epothilone B induced a concentration- and application-dependent antiproliferative effect on the cells, with IC(50) values in the low nanomolar range. Colony forming assays showed a synergistic radiosensitive effect on both cell lines which was dependent on incubation time and applied concentration of epothilone B. The γH2AX assays demonstrated that ionizing radiation combined with the drug resulted in a concentration-dependent increase in the number of double-strand breaks and suggested a reduction in DNA repair capacity. Epothilone B produced enhanced microtubule bundling and abnormal spindle formation as revealed by immunofluorescence microscopy and caused microtubule formation from purified tubulin. The results of this study showed that epothilone B displays cytotoxic antitumor activity at low nanomolar concentrations and also enhances the radiation response in the tumor cells tested; this may be induced by a reduced DNA repair capacity triggered by epothilone B. It was also demonstrated that epothilone B in fact targets microtubules in a more effective manner than paclitaxel.


Involvement of rostral ventromedullar neuronal structures in nitric oxide modulation of central chemosensitive drive.
Surface perfusion of the rostral ventromedullar cerebral subdivisions with artificial cerebrospinal fluid containing exogenous NO donor sodium nitroprusside (0.1 mM) increased the discharge rate of the phrenic nerve and potentiated the response of the respiratory center to hypercapnia in narcotized mature rats. The latter reaction was prevented by blockage of NO-synthase in rostral ventromedullar neural structures with N(omega)-nitro-L-arginine methyl ester (L-NAME, 0.3 mM). It was hypothesized that rostral ventromedullar neural structures are involved in modulatory action of NO on central chemosensitive drive.


The cohort of long-term survivors at the German childhood cancer registry.
With the increasing number of long-term survivors among patients diagnosed with cancer during childhood, questions concerning late effects have become a major research topic. To ascertain late effects, it is necessary to contact former patients. An essential requirement for such studies is a long-term surveillance (LTS) of former childhood cancer patients in their adolescence and their adulthood. The paper describes the role of the German Childhood Cancer Registry (GCCR) in LTS. A cohort of long-term survivors has been built up over the years. The characteristics of this LTS cohort and strategies for further improvement of LTS will be presented. Since 1980 the GCCR systematically ascertains all malignant neoplasms and benign brain tumours in children under the age of 15 years at diagnosis. Participants are followed up actively by the treating hospitals and the clinical study groups in the first years after diagnosis, and by the GCCR thereafter. Late effects are accessed within the Scientific Society for Paediatric Oncology and Haematology (GPOH) of different groups with different focal points. Those groups are the GCCR (secondary malignant neoplasms), LESS (late effects after chemotherapy), RiSK (late effects after radiotherapy), and the working group on quality of life (quality of life and data on life circumstances). Additionally, the GCCR provides logistics for contacting patients during LTS. The LTS is supported by a recent basic publication ("position paper") by the GPOH. Newly diseased cancer cases are reported to the GCCR very completely. The GCCR contains mainly epidemiological data. Accessorily, the GCCR ascertains a minimum of data for each patient which enables population-based studies involving long-term survivors of childhood cancer. Out of 37 291 children diagnosed with cancer between 1980 and 2004, 8 896 died (until spring 2007). From those not deceased, 21 987 (77.4%) can be followed up further (i.e. current address is known). For about 70% of the patients in the LTS cohort, follow-up data are available and not older than 5 years. Our experience shows that about 80% of former childhood cancer patients agree to continued data storage at the GCCR, 4% explicitly refuse their consent, the remaining do not answer. LTS for patients with leukemia and lymphomas is particularly complete, whereas for patients with brain tumours it is less complete. The LTS is considered highly relevant concerning aspects of clinical quality assurance and epidemiological research. The GCCR can guarantee a continuing development and improvement of existing procedures for LTS. The GCCR expects to achieve contacting a high percentage of former childhood cancer patients also in future LTS, even after long periods of time.


Dexamethasone and retinyl acetate similarly inhibit and stimulate EGF- or insulin-induced proliferation of prostatic epithelium.
Prostatic epithelium proliferates in a defined medium consisting of basal medium RPMI1640 containing transferring (1 microgram/ml), EGF (10 ng/ml), and insulin (3.7 micrograms/ml or 0.1 IU/ml). Although neither dexamethasone nor retinyl acetate affected the proliferation of prostatic epithelium in RPMI1640 containing transferrin alone, they modify the mitogenic effect of EGF and insulin. Dexamethasone at 10(-10) M or retinyl acetate at about 3 X 10(-9) M inhibits proliferation stimulated by EGF. Higher concentrations of dexamethasone (10(-8) - 10(-6) M) or retinyl acetate (3 X 10(-8) - 10(-7) M) enhance the mitogenic activity of EGF. Dexamethasone had a similar effect in the presence of insulin. However, retinyl acetate stimulated, but did not significantly inhibit, proliferation in the presence of insulin. These results suggest that both dexamethasone and retinyl acetate, and possibly other glucocorticoids and retinoids, may regulate the proliferation of prostate epithelium by a dose-dependent modification of the activity of insulin and EGF.


A bis-acetonitrile two-coordinate copper(I) complex: synthesis and characterization of highly soluble B(C(6)F(5))(4)(-) salts of [Cu(MeCN)(2)](+) and [Cu(MeCN)(4)](+).
Copper(I)-acetonitrile complexes are exceedingly useful starting materials for the synthesis of copper(I) complexes with polydentate ligands. To extend the utility of such chemistry to solution studies in relatively low-dielectric solvents (i.e., diethyl ether, toluene) and to aid in obtaining products amenable to X-ray diffraction studies, we have recently begun to employ counteranions such as B(C(6)F(5))(4)(-) for bioinorganic studies. Thus, the synthesis of [Cu(MeCN)(4)]B(C(6)F(5))(4) (1) is presented. Its recrystallization from CH(2)Cl(2)/pentane yields the linear, two-coordinate complex [Cu(MeCN)(2)]B(C(6)F(5))(4) (2), whose centrosymmetric X-ray structure shows that its Cu-N distance is significantly shorter than that in other two-coordinate Cu(I) complexes with nitrogen ligands or that in the tetrahedral complex [Cu(MeCN)(4)]ClO(4). Infrared spectroscopy indicates interesting and diagnostically useful differences between the nu(CN) of 1 and 2.


Isocaloric Pair-Fed High-Carbohydrate Diet Induced More Hepatic Steatosis and Inflammation than High-Fat Diet Mediated by miR-34a/SIRT1 Axis in Mice.
To investigate the different effects of isocaloric high-fat diet (HFD) and high-carbohydrate diet (HCD) on hepatic steatosis and the underlying mechanisms, especially the role of microRNA-34a/silent information regulator T1 (SIRT1) axis, C57BL/6J mice (n = 12/group) were isocaloric pair-fed with Lieber-DeCarli liquid diet containing either high fat (HFLD) or high carbohydrate (HCLD) for 16 weeks. As compared to the HFLD fed mice, despite the similar final body weights, HCLD feeding: (1) induced more severe hepatic steatosis; (2) up-regulated hepatic expression of miR-34a accompanied with significant decrease of SIRT1 and nicotinamide phosphoribosyltransferase (NAMPT), SIRT1 activity and phosphorylation of AMPK; (3) up-regulated de novo lipogenesis (DNL) related proteins expression (ACC, SCD1), and down-regulated expressions of miR-122, miR-370 and miR-33; (4) decreased mRNA expressions of genes Cpt1, Pparα and Pgc1α related to fatty acid oxidation; (5) increased hepatic total cholesterol concentration and decreased expression of cholesterol metabolism related genes Abcg5, Abcg8, Abcg11, Cyp7a1 and Cyp8b1; and (6) induced higher hepatic inflammatory response accompanied with significant increased mRNA expressions of Il1β, Tnfα and Mcp1. Thus, isocaloric HCLD feeding induced greater severity in hepatic steatosis and inflammatory response than HFLD feeding, potentially through miR-34a/SIRT1 axis mediated promotion of DNL, inhibition of fatty acid oxidation and cholesterol metabolism.


A new locus (RP31) for autosomal dominant retinitis pigmentosa maps to chromosome 9p.
Retinitis pigmentosa (RP) is a debilitating disease of the retina affecting approximately 1.5 million people worldwide. RP shows remarkable heterogeneity both clinically and genetically, with more than 40 genetic loci implicated, 12 of which account for the autosomal dominant form (adRP) of inheritance. We have recently identified a French Canadian family that presents with early onset adRP. After exclusion of all known loci for adRP, a genome-wide search established firm linkage with a marker from the short arm of chromosome 9 (LOD score of 6.3 at recombination fraction theta=0). The linked region is flanked by markers D9S285 and D9S1874, corresponding to a genetic distance of 31 cM, in the region 9p22-p13.


[Are titanium implants superior to gold implants in the tympanic cavity?].
Gold implants in reconstructive surgery of the middle ear have proved to work sufficiently: That applies to tympanic cavity conditions without signs of chronic inflammation as also to the bacterially infected middle ear. For some years titanium implants are also in the use and established as well. The biocompatibility of titanium, the clinical and the audiological results are satisfactory. When using gold-prostheses in stapes surgery however, the still unsolved problem of occasional deafness has to be considered. The clinical and functional results of middle ear surgery were examined retrospectively. There was no preselection of patients. 53 patients underwent middle ear surgery with titanium prostheses, 42 patients with gold prostheses. Due to their material properties and due to their delicate shape titanium prostheses (PORP, TORP) can be inserted into the tympanic cavity more simply and with better fit. The audiological results of gold and titanium prostheses are comparable. The extruding rate of the gold prostheses is substantial, whereas the extruding rate of titanium is negligible. The functional results of both materials are comparable. Due to the high extruding rates of gold prostheses titanium is clearly superior.


Potential of protein kinase inhibitors for treating herpesvirus-associated disease.
Herpesviruses are ubiquitous human pathogens that establish lifelong persistent infections. Clinical manifestations range from mild self-limiting outbreaks such as childhood rashes and cold sores to the more severe and life-threatening outcomes of disseminated infection, encephalitis, and cancer. Nucleoside analog drugs that target viral DNA replication provide the primary means of treatment. However, extended use of these drugs can result in selection for drug-resistant strains, particularly in immunocompromised patients. In this review we will present recent observations about the participation of cellular protein kinases in herpesvirus biology and discuss the potential for targeting these protein kinases as well as the herpesvirus-encoded protein kinases as an anti-herpesvirus therapeutic strategy.


Involvement of pyruvate dehydrogenase in product formation in pyruvate-limited anaerobic chemostat cultures of Enterococcus faecalis NCTC 775.
Enterococcus faecalis NCTC 775 was grown anaerobically in chemostat culture with pyruvate as the energy source. At low culture pH values, high in vivo and in vitro activities were found for both pyruvate dehydrogenase and lactate dehydrogenase. At high culture pH values the carbon flux was shifted towards pyruvate formate lyase. Some mechanisms possibly involved in this metabolic switch are discussed. In particular attention is paid to the NADH/NAD ratio (redox potential) and the fructose-1,6-bisphosphate-dependent lactate dehydrogenase activity as possible regulatory factors.


Partial Shocks on Cooperative Multiplex Networks with Varying Degrees of Noise.
In many cooperative networks, such as alliance and trade networks, abrupt and intense changes to the state of the system (which we call "shocks"), can substantially change the network. We examine how such shocks affect multiplex networks via an agent-based model, in which agents add, drop, or change ties to increase their utility. At a certain time-point, some agents are "shocked" by changing (increasing or decreasing) the cost associated with tie-formation or tie-maintenance. Our model makes several improvements to previous models, including (a) only a fraction of nodes are shocked to simulate small wars or scattered tariff increases or decreases and (b) agents can make both utility-maximizing decisions and randomly rewire ties to explore the utility landscape. Interestingly, we find that randomly rewiring ties increases the utility of agents, for reasons similar to simulated annealing in physics. Furthermore, we create a novel metric to determine how networks change after a shock and find that the size of a shock and noise significantly changes the network, but only when agents' incentives for tie-formation are sufficiently high. Together, these results suggest that adding more realism to cooperation network models can give nuanced understanding to network shocks.


Implementing stratified care for acute low back pain in primary care using the STarT Back instrument: a process evaluation within the context of a large pragmatic cluster randomized trial.
Although risk-stratifying patients with acute lower back pain is a promising approach for improving long-term outcomes, efforts to implement stratified care in the US healthcare system have had limited success. The objectives of this process evaluation were to 1) examine variation in two essential processes, risk stratification of patients with low back pain and referral of high-risk patients to psychologically informed physical therapy and 2) identify barriers and facilitators related to the risk stratification and referral processes. We used a sequential mixed methods study design to evaluate implementation of stratified care at 33 primary care clinics (17 intervention, 16 control) participating in a larger pragmatic trial. We used electronic health record data to calculate: 1) clinic-level risk stratification rates (proportion of patients with back pain seen in the clinic over the study period who completed risk stratification questionnaires), 2) rates of risk stratification across different points in the clinical workflow (front desk, rooming, and time with clinician), and 3) rates of referral of high-risk patients to psychologically informed physical therapy among intervention clinics. We purposively sampled 13 clinics for onsite observations, which occurred in month 24 of the 26-month study. The overall risk stratification rate across the 33 clinics was 37.8% (range: 14.7-64.7%). Rates were highest when patients were identified as having back pain by front desk staff (overall: 91.9%, range: 80.6-100%). Rates decreased as the patient moved further into the visit (rooming, 29.3% [range: 0-83.3%]; and time with clinician, 11.3% [range: 0-49.3%]. The overall rate of referrals of high-risk patients to psychologically informed physical therapy across the 17 intervention clinics was 42.1% (range: 8.3-70.8%). Barriers included staffs' knowledge and beliefs about the intervention, patients' needs, technology issues, lack of physician engagement, and lack of time. Adaptability of the processes was a facilitator. Adherence to key stratified care processes varied across primary care clinics and across points in the workflow. The observed variation suggests room for improvement. Future research is needed to build on this work and more rigorously test strategies for implementing stratified care for patients with low back pain in the US healthcare system. Trial registration: ClinicalTrials.gov ( NCT02647658 ). Registered January 6, 2016.


Local relapse of prostate cancer after primary definitive treatment: the management.
Prostate cancer (PCa) is the most commonly diagnosed malignancy in men and the second leading cause of cancer-related death in industrialized countries. Even if the healing chances are very high after definitive treatment of localized disease, 20-30% of patients experience recurrence. A review of the literature on the management of local recurrent prostate cancer was conducted using the Medline and Embase electronic databases. Search terms included "biochemical relapse", "PSA recurrence", "prostate cancer", "prostate cancer recurrence", "prostate salvage therapy", "radiorecurrent prostate cancer", "Re-HIFU", "post HIFU", "post cryoablation", "postradiation", and "postprostatectomy salvage". The search was restricted to English-language articles. The websites of guidelines organizations (EAU, AUA, NICE) were consulted in order to identify evidence-based practice guidelines. The present role of salvage prostatectomy and radiation therapy was studied and today's outcomes and tomorrow perspectives of salvage focal therapies as cryoablation and HIFU have been analyzed. Although the treatment landscape for patients with biochemical recurrence prostate cancer remains challenging, new research is helping to identify patient populations suitable for specific therapies. Further evaluation in prospective clinical trials will hopefully confirm the role of therapeutic options in clinical practice and the impact on the long-term survival.


Nuclear medicine studies of the heart.
Nuclear imaging techniques are well established diagnostic tools in clinical cardiology, providing noninvasive information about myocardial perfusion, function and metabolism. The cost-effectiveness of radionuclide imaging in the diagnostic work-up of patients with coronary artery disease has been demonstrated. Additionally, the documented prognostic value of scintigraphic parameters is of clinical importance to guide decision making. Advances in technology, new radiotracers and new applications contribute to continuous growth in the field of nuclear cardiology. Multi-headed gamma camera systems lead to higher spatial resolution and sensitivity of cardiac single photon emission tomography (SPECT), and they also provide the opportunity for attenuation correction or electrocardiographic gating of SPECT images. Objective quantitative values of perfusion, function and metabolism are derived from scintigraphic data by use of improved software and hardware. With the latest developments in tracer technology, imaging of myocardial necrosis, receptor systems and autonomic innervation has become a reality and will lead to new clinical applications in the future.


Allogeneic grafts of fetal dopamine neurons: immunological reactions following active and adoptive immunizations.
To define the importance of adoptive sensitization and duration of graft residence on transplant alloimmunization, behavioral and histochemical parameters were examined in unilaterally 6-OHDA-lesioned F344 rat hosts which received fetal ventral mesencephalic (VM) grafts from Wistar-Furth (WF) donors. In all animals which showed increased rotations after alloimmunization, increased numbers of T cell receptor (TcR) positive, CD8+ lymphocytes were detected in the grafts. In addition, an increased density of class I MHC antigens was seen in the graft and in the adjacent host brain. Lesser numbers of CD4+, CD11b+, and MHCII+ positive elements were also seen. Perivascular cuffing was often found in actively immunized animals. An increase in TcR+ and MHC class I+ elements was also seen in animals only adoptively immunized. The tyrosine hydroxylase positive graft area was also markedly reduced in actively immunized animals and the extent of reduction correlated with the number of cells used for immunization. These studies indicate that established allografts can evade rejection as long as host lymphocytes are not activated against graft alloantigens. In addition, increasing graft residence time in the host and adoptive immunization render the graft more susceptible to subsequent rejection.


An analysis of interactions between three structurally diverse anthocyanidins, as well as their glucosides, and model biological membranes, albumin, and plasmid DNA.
The aim of the study is to investigate the differences in the interaction of three structurally diverse anthocyanidins, namely peonidin, petunidin, and delphinidin, as well as their glucosides with model biological membranes, human albumin, and plasmid DNA in order to look into their structure-activity relationships. Fluorimetric studies, as well as ATR-FTIR analyses, were jointly used in order to determine the changes observed in both the hydrophilic and hydrophobic layers of cell-mimic membranes (MM) which reflected the membrane lipid composition of tumour cells and red blood cell membranes (RBCM). Our results showed that anthocyanins and anthocyanidins can cause an increase in the packing order of the polar heads of lipids, as well as interact with their deeper layers by reducing the fluidity of lipid chains. The results presented here indicate that all compounds tested here possessed the ability to bind to human serum albumin (HSA) and the presence of a glucose molecule within the structures formed by anthocyanidin reduces their ability to bind to proteins. Using fluorescence correlation spectroscopy, it was demonstrated that the compounds tested here were capable of forming stable complexes with plasmid DNA and, particularly, strong DNA conformational changes were observed in the presence of petunidin and corresponding glucoside, as well as delphinidin. The results we obtained can be useful in comprehending the anthocyanins therapeutic action as molecular antioxidants and provide a valuable insight into their mechanism of action.


Optimized radiofrequency lesions through local impedance guidance for effective CTI ablation in right atrial flutter.
Although radiofrequency (RF) catheter ablation of cavo-tricuspid isthmus (CTI) is an established treatment for typical right atrial flutter (RAFL), it remains to be established whether local tissue impedance (LI) is able to predict effective CTI ablation and what LI drop values during ablation should be used to judge a lesion as effective. We aimed to investigate the ability of LI to predict ablation efficacy in patients with RAFL. RF delivery was guided by the DirectSense™ algorithm. Successful single RF application was defined according to a defragmentation of atrial potentials (DAP), reduction of voltage (RedV) by at least 80% or changes on unipolar electrogram (UPC). The ablation endpoint was the creation of bidirectional conduction block (BDB) across the isthmus. 392 point-by-point RF applications were analyzed in 48 consecutive RAFL patients. The mean baseline LI was 105.4 ± 12Ω prior to ablation and 92.0 ± 11Ω after ablation (p < 0.0001). According to validation criteria, absolute drops in impedance were larger at successful ablation sites than at ineffective ablation sites (DAP: 17.8 ± 6Ω vs. 8.7 ± 4Ω; RedV: 17.2 ± 6Ω vs. 7.8 ± 5Ω; UPC: 19.6 ± 6Ω vs. 10.1 ± 5Ω, all p < 0.0001). LI drop values significantly increased according to the number of criteria satisfied (ranging from 7.5Ω to 19.9). BDB was obtained in all cases. No procedure-related adverse events were reported. A LI-guided approach to CTI ablation was safe and effective in treating RAFL. The magnitude of LI drop was associated with effective lesion formation and BDB and could be used as a marker of ablation efficacy. Catheter Ablation of Arrhythmias with a High-Density Mapping System in Real-World Practice (CHARISMA). URL: http://clinicaltrials.gov/ Identifier: NCT03793998.


Changes in chemical composition and collagen structure of dentine tissue after erbium laser irradiation.
Erbium laser radiation has a great affinity for the water molecule, which is present in quantity in biological hard tissues. The objective of this work is to identify chemical changes by infrared spectroscopy of irradiated dentine by an Er:YAG-2.94 microm laser. The irradiation was performed with fluences between 0.365 and 1.94 J/cm2. For the infrared analysis a Fourier transform infrared spectrometer was used. After the irradiation were observed: loss of water, alteration of the structure and composition of the collagen, and increase of the OH- radical. These alterations can be identified by a decrease in intensity of the water band between 2800-3800 cm(-1), OH- band at 3575 cm(-1) and bands ascribed to organic matrix between 2800-3400 cm(-1) and 1100-1400 cm(-1).


2-Methoxyestradiol, an endogenous metabolite of 17beta-estradiol, inhibits adipocyte proliferation.
The effects of 2-methoxyestradiol (2ME), a naturally occurring mammalian metabolite of 17beta-estradiol, on adipocyte growth has been investigated in mouse brown adipose tissue precursor cells developed in primary culture. 2ME inhibits brown adipocyte proliferation in a dose-response manner (IC50 = 1.7 x 10(-6) M for DNA synthesis), with much higher potency than its hormone precursor 17beta-estradiol, and cells acquire the typical differentiated morphology--more round with a higher content of triglycerides. 2ME causes similar effects in the immortal brown adipocyte tumor-derived hibernoma cell line HIB 1B and the immortal 3T3-F442A white adipocyte line. These findings suggest a possible role for 2ME in adipocyte proliferation, and probably in the differentiation process, entering the cells in the adipogenic program.


The Response of Susceptible and Pyrethroid-Resistant Blattella germanica (Dyctioptera: Blattellidae) to Shelter-Associated Cues.
In this work, it was studied the role of faeces in the location and permanence in a shelter in susceptible and pyrethroid-resistant individuals of Blattella germanica (Linnaeus 1767). Additionally, the effect of different concentrations of palmitic acid on the modulation of these behaviours was tested. A shelter constituted by a square cardboard structure was offered to susceptible as well as to resistant specimens. The shelter bases were treated with faecal extracts obtained from susceptible or resistant cockroaches, or with solutions of palmitic acid. The behaviour of susceptible as well as resistant specimens was analysed using infrared videography software. Susceptible's faecal extract attracted both specimens since the time spent by cockroaches to locate the treated shelters was lower, whereas the faecal extract from resistant insects did not elicit any effect on both strains. Faecal extracts showed an arrestant effect on both strains, suggested by the time spent inside the shelter that was significantly higher in their presence. On the other hand, treatment with palmitic acid produced an attractant or a repellent effect depending on the concentration and strain. The tested lower concentration was attractant to susceptible insects, but did not produce any effect on resistant ones. In addition, the higher concentrations did not produce any effect on susceptible individuals, but resulted repellent for resistant ones. Palmitic acid did not produce an arrestant effect on the strains as there was not an increase in time spent inside the shelter in the presence of this substance. An increase in the number of visits to the shelter and to the periphery was also observed in shelters treated with the faecal extract and with the lower concentration of palmitic acid. These results show that compounds of the susceptible faeces were attractant to cockroaches of both strains, while faecal extracts from resistant insects were not. Moreover, a dual effect of palmitic acid was observed, being attractant at low concentrations and repellent as concentration increased. Additionally, a difference in the concentration threshold at which the effect of this substance changes was observed between strains.


Staging and monitoring in the treatment of lymphomas.
Lymphoma staging systematically includes a CT scan of the cervical, thoracic and abdominopelvic regions. PET is indicated in diffuse large B cell lymphomas (DLBCL) and Hodgkin's disease. Evaluation of the response to treatment is based on Cheson's 1999 morphological criteria, which have been replaced by the 2007 IWC criteria, which combine morphological and metabolic responses. CT and FDG-PET are complementary in characterizing residual masses: if negative, a PET scan indicates the absence of residual disease, if positive; it directs a CT-guided biopsy to obtain the histological evidence. Monitoring clinical features and laboratory values is primordial following treatment. Imaging is performed as a second intention for investigating a relapse, if necessary associated with a PET scan. Multimodal imaging implies multidisciplinary consultation between haematologists, imaging specialists and histopathologists.


[Treatment results with weak concentrations of pilocarpine in patients with primary glaucoma (Spersacarpin 0.25%) (author's transl)].
25 Patients suffering from simple chronic glaucoma were treated with a 0.25% Spersacarpin solution. The intraocular tension was normalised in 22 Patients; only in 3 was it necessary to use stronger concentrations. The Spersacarpin 0.25% solution, which has a strong hypotensive and myotic effect but hardly any myopia inducing action, was well tolerated by all patients.


Correction of aortic insufficiency with an external adjustable prosthetic aortic ring.
Less invasive, valve-sparing options are needed for patients with aortic insufficiency (AI). We sought to evaluate the feasibility of reducing AI with an external adjustable aortic ring in an ovine model. To create AI, five sheep underwent patch plasty enlargement of the aortic annulus and root by placement of a 10 x 15 mm pericardial patch between the right and noncoronary cusps. An adjustable external ring composed of a nylon band was fabricated and placed around the aortic root. Aortic flow, aortic pressure, and left ventricular pressures were measured with the ring loose (off) and tightened (on). Mean regurgitant orifice area decreased by 86%, from 0.07 +/- 0.03 cm2 (ring loose, off) to 0.01 +/- 0.00 cm2 (ring tightened, on) [p < 0.01]. The regurgitant fraction decreased from 18 +/- 4% to 2 +/- 1% [p < 0.01]. The ring did not significantly affect stroke volume and aortic pressure. An ovine model of aortic root dilatation resulting in acute AI has been developed. In this model, application of an external, adjustable constricting aortic ring eliminated AI. An aortic ring may be a useful adjunct in reducing AI secondary to annular dilatation.


Frontal Assessment Battery is a marker of dorsolateral and medial frontal functions: A SPECT study in frontotemporal dementia.
The objective of this study is to identify the cerebral regions that are assessed by the Frontal Assessment Battery (FAB). Using SPM voxel-based analysis, we looked for correlations between FAB performance and brain SPECT perfusion in 47 patients with the frontal variant of frontotemporal dementia (fv-FTD) recruited by the French FTD research network, a multicentre initiative of French University hospitals with expertise in the field of dementia. A significant correlation was found between FAB performance and perfusion in the medial and dorsolateral frontal cortex bilaterally, independently of age, gender and MMSE. No correlations were observed with orbital frontal or parietal perfusion, in spite of the presence of hypoperfusion in these areas, or with perfusion of any other cortical or subcortical region. These findings confirm that the FAB is an adequate tool for assessing functions related to the dorsolateral and medial frontal cortex, and is thus useful for the evaluation of diseases associated with frontal dysfunction.


Students' perceptions of teaching factors that demotivate their learning in lectures and laboratory-based skills practice.
The objective of this paper is to report students' perceptions of factors that demotivated their learning in lectures and laboratory-based skills practice settings. A total of 23 students were recruited from the Bachelor of Nursing courses at three Japanese universities, using purposive sampling. A semi-structured interview was conducted with each participant between November 2017 and January 2018 to elicit their perceptions about which aspects of the teaching context demotivated their engagement in learning. The results were analyzed using thematic analysis. Three themes were generated: a restrictive environment, discouraging attitudes and discouraging teaching approaches. To prevent students from experiencing demotivation, teachers in the nursing faculty need to manage learning resources more effectively, create a quiet and focused atmosphere to allow students to concentrate, and be enthusiastic about teaching. They also need to add value to their classes, help students to follow lectures, and ensure that the workload they give their students is appropriate.


Electrophysiologic studies in patients with bundle branch block.
A total of 401 patients with chronic bundle branch block underwent electrophysiologic studies and were followed for a mean of approximately 30 months. Patients with an infranodal conduction time (H-Q) greater than or equal to 70 ms had a significantly higher incidence of progression to spontaneous second-degree or third-degree atrioventricular (AV) block (12%) compared with those with H-Q less than 70 ms (3.5%). The incidence of AV block was 25% for those with H-Q greater than or equal to 100 ms. Although the incidences of all deaths and cardiac deaths were higher for the H-Q greater than or equal to 70 ms group, there was no statistically significant difference in the incidence of sudden deaths in the two groups. A subgroup of 77 patients underwent prophylactic pacemaker insertion based on the presence of transient neurologic symptoms and/or a prolonged H-Q interval. We found no significant difference in the incidence of relief of symptoms or incidence of cardiac or sudden deaths between paced and unpaced groups. A subgroup of 25 patients with syncope underwent ventricular stimulation studies and ventricular tachycardia was induced in 14. We conclude that a markedly prolonged H-Q interval (greater than or equal to 100 ms) may be of value in detection of spontaneous AV block. Prophylactic pacing is of no value in either relief of symptoms or prolongation of life. Complete electrophysiologic studies, including ventricular stimulation studies, are indicated for patients with bundle branch block and syncope.


Distal interphalangeal joint arthrodesis with the Herbert headless compression screw: outcomes and complications in 64 consecutively treated joints.
Arthrodesis of the distal interphalangeal (DIP) joint is indicated for the treatment of arthritis. While several techniques have been recommended, the use of headless compression screws has grown in popularity. Rates of union reported vary widely, ranging from 80% to 100%, with most studies based on small series. The purpose of this study was to review the outcomes and complications associated with DIP joint arthrodesis using the Herbert headless compression screw in a large case series. The medical charts, surgical reports, and X-rays for patients undergoing DIP joint arthrodesis with a Herbert screw between January 1996 and May 2006 were retrospectively reviewed to determine the frequency and types of complications. All operations were performed by the senior author at a single institution. Of 64 joints in 51 patients that were treated with the Herbert screw, a total of 95% (n = 61) went on to union. Union within 3 months occurred in 89% (n = 57) while delayed union (between 3 and 6 months) occurred in 6% (n = 4). Nonunion requiring subsequent revision arthrodesis occurred in 5% (n = 3). Screw removal for symptomatic hardware was required in 8% (n = 5). Fusion of the DIP joint with the Herbert screw can be achieved at rates that are comparable to other techniques and other headless compressive screws. However, while complications do occur, the Herbert screw provides an acceptable rate of union and ease of operative technique, making it a suitable procedure for DIP joint arthrodesis.


Diagnostic value of percutaneous transhepatic cholangiography judged by personal experience of 58 patients.
During the last seven years percutaneous transhepatic cholangiography (PTC) using a fine needle was performed in 58 patients. The biliary tract was visualised and opacified in the 52 patients with dilated biliary ducts. The correct diagnoses were made in 51 of them. In six patients with a non-dilated biliary system, successful PTC was carried out with the correct diagnoses in four of them. There were four non-fatal complications; two septic reactions, one haemobilia, and one subcapsular haematoma, but all were controlled by conservative treatment alone. The PTC with a fine needle is an accurate, useful and safe procedure of choice for the diagnosis of obstructive jaundice and jaundice of obscure origin. It is useful in distinguishing hepatocellular jaundice from obstructive jaundice thus preventing an unnecessary operation. Failure to visualise the biliary tract by an experienced hand strongly indicates that the jaundice is non-obstructive.


Quantitative split dose thallium-201 imaging with exercise: a technique for obtaining rest and exercise perfusion images in one setting and markedly reducing the study time.
In the current study, a technique for performing serial thallium imaging after two separate tracer injections was applied to exercise thallium imaging, thus allowing the acquisition of rest and exercise images within 1 hour. Twenty-four patients with and 10 patients without significant coronary artery disease were studied. One mCi of thallium-201 was injected intravenously and imaging was performed at rest in three projections. The patient was then stressed and an additional 1 mCi of thallium injected during exercise. Images in the same three projections were collected. After computer realignment, the rest image was subtracted from the exercise image to produce an image representing perfusion during exercise. All 24 patients with coronary artery disease had a positive study, while 9 of 10 without disease had a negative study. The images were then interpreted using a computer method designed to quantitate regional myocardial thallium distribution and redistribution. With quantitative interpretation, 23 of 24 patients with coronary disease had a positive study, while only 1 without disease had a positive study. With qualitative interpretation, 39 (89%) of 44 stenosed coronary arteries demonstrated thallium defects in corresponding myocardial segments, while 54 (93%) of 58 nonstenosed coronary arteries did not. With quantitative interpretation, 38 (86%) of 44 stenosed coronary arteries demonstrated thallium defects in corresponding myocardial segments, while 53 (91%) of 58 nonstenosed coronary arteries did not. A split dose thallium imaging technique that allows imaging before and immediately after exercise, thus markedly reducing the study time, has been validated.


Efficient and clean preparation of rare prosaikogenin D by enzymatic hydrolysis of saikosaponin B2 and response surface methodology optimization.
Prosaikogenin D, a rare secondary saponin in Radix Bupleuri, has much higher in vivo bioactivities than its original glycoside saikosaponin B2. Its preparation methods, such as conventional acid hydrolysis and column chromatograph, are unfriendly to environment with serious pollution and undesired products. The aim of this study was to establish an efficient and clean approach for convenient preparation of this rare steroid saponin based on the enzymatic hydrolysis. Cellulase was selected from four commercial enzymes due to its higher hydrolysis performance. Then the hydrolysis conditions were optimized by response surface methodology after preliminary investigation on affecting factors by single-factor experiments. The reaction system was constructed by 100 μg/mL of saikosaponin B2 and 8.00 mg/mL of cellulase, which was incubated in HAc-NaAc buffer (pH 4.7) at 60 °C for 33 h. Consequently, a high conversion ratio of the substrate has been achieved at 95.04 %. The newly developed strategy is an efficient and clean approach for the preparation of prosaikogenin D and it is a promising technology in industrial application.


Identifying Cyclin A/Cdk1 Substrates in Mitosis in Human Cells.
Cyclin A promotes Cdk activity in a cell cycle-dependent manner to facilitate specific cell cycle events and transitions with an established role for DNA replication in S phase. Recent evidence demonstrates that cyclin A also activates Cdk during early mitosis to promote faithful chromosome segregation by regulating the stability of kinetochore-microtubule (k-MT) attachments. Here we describe a methodology to identify protein substrates of cyclin A/Cdk during mitosis in human cells. The method combines selective cell cycle synchrony in mitosis with stable isotope labeling of amino acids in cell culture (SILAC) coupled to mass spectrometry. This strategy identified a catalogue of potential cyclin A/Cdk substrates in mitosis, as well as unveiled potential intersections between signaling regulated by Aurora, Polo-like, and Cdk mitotic kinases.


Sporadic SNCA mutations A18T and A29S exhibit variable effects on protein aggregation, cell viability and oxidative stress.
α-synuclein aggregation is the hallmark feature of Parkinson's disease. Both familial and sporadic forms of the disease exhibit this feature. Several mutations have been identified in patients and are associated with the disease pathology. We have used site-directed mutagenesis to generate α-synuclein mutant variants tagged with GFP. Fluorescence microscopy, flow cytometry, western blotting, cell viability and oxidative stress analysis were performed to investigate the effect of two less studied α-synuclein variants. In this study we characterized two less studied α-synuclein mutations, A18T and A29S, in the well-established yeast model. Our data shows variable expression, distribution and toxicity of the protein in the mutant variants A18T, A29S, A53T and WT. The cells expressing the double mutant variant A18T/A53T showed the most increase in the aggregation phenotype and also depicted reduced viability suggesting a more substantial effect of this variant. The outcome of our study highlights the variable localization, aggregation phenotype and toxicity of the studied α-synuclein variants. This underscores the importance of in-depth analysis of every disease-associated mutation which may result in variable cellular phenotype.


Antidiarrhoeal Activity of Hydroethanolic Leaf Extract of Bryophyllum pinnatum Lam. Kurtz (Crassulaceae).
Bryophyllum pinnatum Lam. Kurtz (Crassulaceae) is used in traditional African medicine in the treatment of diarrhoea. To investigate the antidiarrhoeal action of the hydroethanolic leaf extract of Bryophyllum pinnatum (BP). Normal intestinal transit, castor oil-induced intestinal transit, castor oil-induced diarrhoea, gastric emptying and enteropooling models in rodents were used to investigate antidiarrhoeal effect. The possible mechanism of antidiarrhoeal activity was investigated using prazosin (1 mg/kg, s.c; α1, adrenoceptor antagonist), yohimbine (1 mg/kg, s.c; α2 adrenoceptor antagonist), propranolol (1 mg/kg, i.p; α- adrenoceptor non-selective antagonist), atropine (1 mg/kg, s.c; muscarinic cholinergic antagonist), pilocarpine (1 mg/kg, s.c; muscarinic cholinergic agonist), and isosorbide dinitrate (IDN) (150 mg/kg, p.o; nitric oxide donor). BP (25-100 mg/kg, p.o) produced dose-dependent and significant (P < 0.001) decrease in intestinal propulsion in normal and castor oil-induced intestinal transit models in comparison to distilled water (10 ml/kg, p.o.) treated control. This antidiarrhoeal effect was inhibited by propranolol pretreatment but yohimbine, prazosin, or atropine pretreatment failed to block this effect. BP treatment reduced the increased peristaltic activity induced by pilocarpine, however, co-treatment with IDN significantly (P < 0.001) enhanced the antidiarrhoeal effect of the extract. In castor oil-induced diarrhoea test, the extract produced a dose-dependent and significant (P < 0.001) increase in onset of diarrhoea, decreased diarrhoea score, the number and weight of wet stools when compared to control. The in vivo antidiarrhoeal index (ADI(in) vivo)) of 53.52 produced by the extract (50 mg/kg, p.o.) was similar to 76.28 ADI(in vivo) produced by morphine (10 mg/kg, s.c.). The extract produced dose- dependent and significant (P < 0.05; P < 0.001) decrease in the weight and volume of intestinal content in the intestinal fluid accumulation model. In gastric emptying test, BP treatment reduced the quantity of test meal emptied in 1 h but not significant. The results showed that the hydroethanolic leaf extract of Bryophyllum pinnatum possesses antidiarrhoeal activity possibly mediated by interaction with β adrenoceptor, muscarinic cholinergic receptor and nitric oxide pathway.


Alpha neoprotein molecules in normal lenses from animals of different ages and in cataractous lenses.
Alpha neoprotein molecules are formed by an association of A and B subunits resulting in a different quaternary structure than in alpha crystallin. The content of alpha neoprotein vs. alpha crystallin was estimated separately on the crystallin, cryoprotein and albuminoid fractions in lenses of animals of different species and ages, in lenses with experimentally induced cataracts and in human cataractous lenses. Alpha crystallin was determined on immunoadsorbents with bound antibodies restricted to quaternary determinants of this protein. Alpha neoprotein molecules were determined in the filtrate containing lens proteins not bound on the above immunoadsorbent. The filtrate was applied on an anti-A chain immunoadsorbent, and after desorption the A chain-containing molecules were applied on an anti-B chain immunoadsorbent. The amount of lens proteins bound to the latter immunoadsorbent gave a measure of molecules formed by an association of A with B chains other than in alpha crystallin, that is of alpha neoproteins. No alpha neoprotein was present in lenses from 2-week-old rats or in the 3- to 6-month-old calf lens cortex. The 2-year-old bovine lens nucleus did contain alpha neoproteins, but only in the albuminoid fraction. The lenses of adult rats (i.e. 3- to 4-month-old) contained alpha neoprotein molecules mainly in the albuminoid fraction; small amounts were also found in the cryoprotein fraction. All three fractions of lens proteins from 1-year-old rats contained alpha neoprotein molecules. No significant differences in the content of alpha neoprotein molecules were found in lenses with experimentally induced X-ray and galactose cataracts and in normal controls. This finding does not exclude possible structural differences between normal and cataractous alpha neoproteins. In human senile cataractous lenses, the content of alpha neoprotein molecules in all fractions analyzed equaled or exceeded the content of alpha crystallin present. The present findings demonstrate an age-dependent formation in the mammalian lens of alpha crystallin neoproteins lacking native quaternary determinants. The data obtained point to the possibility that the structural lability of alpha crystallin may be a contributing factor in cataractogenesis.


Long-term effect of lamivudine treatment on the incidence of hepatocellular carcinoma in patients with hepatitis B virus infection.
Nucleotide analogues have recently been approved for the treatment of patients with hepatitis B virus (HBV) infection. However, it is still controversial whether the decrease of HBV-DNA amount induced by treatment with nucleotide analogues can reduce the risk of hepatocellular carcinoma (HCC) development in HBV patients. A total of 293 HBV patients without HCC who were treated with lamivudine (LAM) were enrolled in a multicenter trial. The incidence of HCC was examined after the start of LAM therapy, and the risk factors for liver carcinogenesis were analyzed. The mean follow-up period was 67.6 ± 27.4 months. On multivariate analysis for HCC development in all patients, age ≥50 years, platelet count <14.0 × 10(4)/mm(3), cirrhosis, and median HBV-DNA levels of ≥4.0 log copies/ml during LAM treatment were significant risk factors. The cumulative carcinogenesis rate at 5 years was 3% in patients with chronic hepatitis and 30% in those with cirrhosis. For the chronic hepatitis patients, the log-rank test showed the significant risk factors related to HCC development to be age ≥50 years, platelet count <14.0 × 10(4)/mm(3), and hepatitis B e antigen negativity, but median HBV-DNA levels of <4.0 log copies/ml (maintained viral response, MVR) did not significantly suppress the development of HCC. In cirrhosis patients, however, the attainment of MVR during LAM treatment was revealed to reduce the risk of HCC development. These results suggest that the incidence of HCC in HBV patients with cirrhosis can be reduced in those with an MVR induced by consecutive LAM treatment.


Experimental biomechanical study of the primary stability of different osteosynthesis systems for mandibular reconstruction with an iliac crest graft.
We did biomechanical loading tests to compare the quantity and quality of interfragmentary movement in fractured human cadaver mandibles that had been reconstructed with iliac crest using 3 different osteosynthesis systems. Eighteen mandibles from human cadavers with a 4.5 cm paramedian L-type defect were reconstructed with bone from the iliac crest using 3 different osteosynthesis systems and continuously loaded on the "Mandibulator" test bench. Six mandibles each had the bones joined together using 2 monocortical non-locking plates, 2 monocortical locking plates, or a single bicortical locking plate/fracture gap. Macroscopic deformation, failure mechanisms, and movement of the fracture gap in all 3 dimensions were assessed and quantified over increasing loading by PONTOS(®) optical measurement systems. Final mechanisms of failure were excessive deformation of the plate, fracture of the mandibular fragments, and failure of the iliac crest graft. The plate became deformed mainly in the miniplate group. The iliac crest graft failed in all the specimens in which osteosynthesis was performed by a 6-hole TriLock(®) plate. Interfragmentary movement was minimised in the miniplate group. All three osteosynthesis systems provided sufficient stability for reconstruction when mechanically loaded up to 100 N. The miniplate allowed less movement in the gap and gave better stability than the two TriLock(®) plate systems. The superiority of the miniplate was significant when compared with the 4-hole TriLock(®) plate. The transplant failed mainly in the 6-hole TriLock(®) group, which suggests that the iliac crest graft works better with the miniplate as a more malleable osteosynthesis system.


Conformational analysis of helical poly(beta-L-aspartate)s by IR dichroism.
Poly(beta-l-aspartate)s are known to take up helical conformations reminiscent of the alpha-helix of polypeptides. The isobutyl, n-butyl, and 2-methoxyethyl esters have been examined by polarized ir spectroscopy in order to discriminate between the left (1L) and right (2R)-handed conformations, which are known to be compatible with the 13/4-helix adopted by these polyamides when crystallized in the hexagonal form. Dichroic ratios obtained from samples stretched in poly(ethylene oxide) together with orientation measurements made by x-ray diffraction were used to estimate the transition moment directions of amide A, I, and II bands with respect to the fiber axis. These were compared to those calculated by modeling simulations to conclude that the right-handed conformation consisting of 14-membered hydrogen-bonded rings is the correct model for the 13/4-helix. These results give definite support to earlier molecular mechanics calculations, which had shown that the 2R model is energetically favored over the 1L by about 2.5 kcal/(mol residue).


Value of preoperative imaging in the diagnostics of isolated metopic suture synostosis: a risk-benefit analysis.
Radiographic evaluation including plain radiographies and computed tomographic (CT) scans are considered as a necessary tool for diagnosis of craniosynostosis. As recently concerns about harmful effects of ionising radiation in children have been raised, some authors have suggested the use of magnetic resonance imaging (MRI) as a helpful alternative in preoperative imaging of patients with isolated metopic synostosis. Besides confirming the diagnosis of trigonocephaly, MRI is the superior technique for the evaluation of underlying brain anomalies. However, if the benefit of preoperative imaging justifies possible side effects is still discussed controversially. Hence, this study investigated the value of preoperative imaging for the diagnosis of isolated synostosis of the metopic suture compared to a sole clinical examination. In a series of 63 cases with isolated metopic craniosynostosis operated at the Department of Oral and Maxillofacial Surgery, 48 (76.2%) patients received additional radiography or MRI investigation, while in 15 (23.8%) patients the diagnosis was based on clinical examinations only. In all patients, diagnosis was confirmed intra-operatively by a fused metopic suture. CT scans with three-dimensional reconstruction (12.5%) or plain radiographs (39.6%) did not provide any additional benefit for the diagnosis or the surgical treatment. In 23 patients (47.9%), MRI showed the typical soft-tissue alterations like triangular brain deformation in the frontal area. Besides these findings, no brain or other underlying anomalies were diagnosed which had required any additional treatment. The incidence of underlying brain abnormalities in isolated metopic synostosis seemed not to be different from that of the general population. As the characteristic clinical manifestations were sufficient for an accurate diagnosis of isolated metopic synostosis, and with respect to the biological effects of ionising radiation and risks of sedation especially in infants, preoperative imaging should be reduced to a minimum.


Fourier ptychographic reconstruction using Poisson maximum likelihood and truncated Wirtinger gradient.
Fourier ptychographic microscopy (FPM) is a novel computational coherent imaging technique for high space-bandwidth product imaging. Mathematically, Fourier ptychographic (FP) reconstruction can be implemented as a phase retrieval optimization process, in which we only obtain low resolution intensity images corresponding to the sub-bands of the sample's high resolution (HR) spatial spectrum, and aim to retrieve the complex HR spectrum. In real setups, the measurements always suffer from various degenerations such as Gaussian noise, Poisson noise, speckle noise and pupil location error, which would largely degrade the reconstruction. To efficiently address these degenerations, we propose a novel FP reconstruction method under a gradient descent optimization framework in this paper. The technique utilizes Poisson maximum likelihood for better signal modeling, and truncated Wirtinger gradient for effective error removal. Results on both simulated data and real data captured using our laser-illuminated FPM setup show that the proposed method outperforms other state-of-the-art algorithms. Also, we have released our source code for non-commercial use.


"It's Not an Acting Job … Don't Underestimate What a Simulated Patient Does": A Qualitative Study Exploring the Perspectives of Simulated Patients in Health Professions Education.
Simulated patients (SPs) are individuals who have learned to realistically portray patient roles in health professional education. Program recommendations are increasing for simulation programs, and as key stakeholders, SPs' perspectives seem underrepresented. The aim of the study was to explore the experiences, perspectives, and practices of SPs to gain insights on topics of importance to SPs and inform program recommendations. An interpretivist research paradigm and qualitative design were adopted. Eighteen SPs participated in 2 focus groups that were audio recorded, transcribed, and deidentified. Three researchers completed inductive thematic analysis. Institutional ethical approval was obtained. Three themes represented the different elements of SP practice: becoming and being a SP, preparing for a SP role, and performing a SP role. Simulated patients identify as educated specialists with unique responsibilities and attributes. Simulated patients are committed to representing the perspectives of real patients, while simultaneously supporting learners and educators. Simulated patients can feel unprepared to perform a role but have innovated responsive strategies. Simulated patients considered 3 primary aspects to their practice and shared ways that they might be well supported. Simulated patients represent a community of practice, characterized by mutual engagement, joint enterprise, and a shared repertoire. Ongoing SP input in SP programs may benefit SPs and lead to higher-quality educational experiences for learners.


Increased expression of glycolysis-associated genes in oncogene-transformed and growth-accelerated states.
An accelerated rate of glucose transport and catabolism is a common characteristic of cellular transformation. We have previously found elevated expression of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in human pancreatic and colonic adenocarcinomas (Schek et al.: Cancer Res 48:6354-6359, 1988). To investigate further the expression of this enzyme in the process of tumorigenesis, we examined GAPDH expression in a panel of oncogene-transformed fibroblasts. Significant elevations of GAPDH mRNA and glucose transporter protein mRNA levels were observed in ras- and mos-transformed NIH 3T3 cells, whereas little or no change was found in c-src-, v-src-, c-myc-, E1A-, v-fos-, and PKC-gamma-transfected cells. Furthermore, the level of GAPDH mRNA correlated with the transformed state in a series of ras-transformed and revertant cell lines. Immunoblot analysis confirmed that GAPDH polypeptide was significantly elevated in the cell lines with elevated mRNA levels. Cell cycle analysis data suggested that the effect on GAPDH expression correlated with oncogene expression rather than cell growth fraction. These results suggest that altered GAPDH gene expression occurs during some growth deregulated states, and this, along with increased glucose transporter (and possibly other glycolytic enzyme) expression, is likely to contribute to the increased metabolic capacity of cells in these states.


Producing reverse phase protein microarrays from formalin-fixed tissues.
In most hospitals around the world FFPE (formalin fixed, paraffin embedded) tissues have been used for diagnosis and have subsequently been archived since decades. This has lead to a sizeable pool of this kind of tissues. Till quite recently it was not possible to use this congeries of samples for protein analysis, but now several groups described successful protein extraction from FFPE tissues. In this chapter, we describe a protein extraction protocol established in our laboratory combined with the use of reverse phase protein microarray.


Effects of urinary macromolecules on the crystallization of calcium oxalate.
The macromolecular fraction of urine with a molecular weight above 3,000 was isolated by dialysis. In the dialysed urine the rate of calcium oxalate (CaOx) crystallization was reduced both in the presence and absence of CaOx seed crystals. There was a clear relationship between crystallization and the relative concentration of the dialysed urine, with the highest crystallization propensity at the lowest concentration of macromolecules. Dilution of dialysed urine also affected crystal size distribution, with a predominance of small (2.8-4.5 microns) crystals in 100% dialysed urine and of large (5.6-14.0 microns) crystals in 5% dialysed urine. This is consistent with a macromolecular inhibition of both crystal growth and aggregation. Analysis of the crystal size distribution 120 min after supersaturation of whole urine to a level at which approximately 100 crystals in the size interval 3.5-5 microns were detected in a Coulter counter surprisingly disclosed a higher mean crystal volume in urine samples from normal subjects than from stone formers. This gives support to the assumptions that macromolecules might be of importance during the initial phase of CaOx crystallization and that urine from stone formers and normal subjects might be different in this respect.


Sex as a prognostic factor in adult-type diffuse gliomas: an integrated clinical and molecular analysis according to the 2021 WHO classification.
To investigate whether type-specific sex differences in survival exist independently of clinical and molecular factors in adult-type diffuse gliomas according to the 2021 World Health Organization (WHO) classification. A retrospective chart and imaging review of 1325 patients (mean age, 54 ± 15 years; 569 females) with adult-type diffuse gliomas (oligodendroglioma, IDH-mutant, and 1p/19q-codeleted, n = 183; astrocytoma, IDH-mutant, n = 211; glioblastoma, IDH-wildtype, n = 800; IDH-wildtype diffuse glioma, NOS, n = 131) was performed. The demographic information, extent of resection, imaging data, and molecular data including O6-methylguanine-methyltransferase promoter methylation (MGMT) promotor methylation were collected. Sex differences in survival were analyzed using Cox analysis. In patients with glioblastoma, IDH-wildtype, female sex remained as an independent predictor of better overall survival (hazard ratio = 0.91, P = 0.031), along with age, histological grade 4, MGMT promoter methylation status, and gross total resection. Female sex showed a higher prevalence of MGMT promoter methylation (40.2% vs 32.0%, P = 0.017) but there was no interaction effect between female sex and MGMT promoter methylation status (P-interaction = 0.194), indicating independent role of female sex. The median OS for females were 19.2 months (12.3-35.0) and 16.2 months (10.5-30.6) for males. No sex difference in survival was seen in other types of adult-type diffuse gliomas. There was a female survival advantage in glioblastoma, IDH-wildtype, independently of clinical data or MGMT promoter methylation status. There was no sex difference in survival in other types of adult-type diffuse gliomas, suggesting type-specific sex effects solely in glioblastoma, IDH-wildtype.


Associations of urinary metal levels with serum hormones, spermatozoa apoptosis and sperm DNA damage in a Chinese population.
Exposure to metals, including essential and nonessential elements, is widespread and may be associated with male reproductive health. To examine whether environmental exposure to metals contributes to reproductive hormone changes, spermatozoa apoptosis and sperm DNA damage in a Chinese population. Eighteen metals (aluminum, arsenic, antimony, chromium, cobalt, copper, cadmium, iron, lead, manganese, molybdenum, nickel, selenium, tin, tungsten, thallium, uranium and zinc) were analyzed in two urine samples collected a few hours apart from male partners of couples attending an infertility clinic. Multivariable linear regression models were used to assess the cross-sectional associations of average urinary metal levels with serum hormones (n=511), spermatozoa apoptosis measures (n=460) and sperm DNA damage parameters (n=516). We found significant inverse dose-dependent trends of urinary tin quartiles with total testosterone (T), and tin, nickel, zinc and molybdenum with the ratio of total T to luteinizing hormone (total T/LH ratio) (all Ptrend<0.05). Additionally, we found significantly dose-dependent trends of increasing urinary manganese quartiles with increasing percentage of Annexin V+/PI- spermatozoa and increasing iron with decreasing percentage of PI+ spermatozoa (both Ptrend<0.05). These dose-dependent trends remained suggestive or significant after controlling for multiple testing and other metals, and they persisted when the metals were modeled as continuous variables in a cubic spline analysis. There were no significant associations between urinary metals and sperm DNA damage after adjustment for multiple testing. Environmental exposure to tin, nickel, zinc and molybdenum may be associated decreased total T or total T/LH ratio; manganese may induce spermatozoa apoptosis, while iron may be important for living spermatozoa. However, additional prospective research is needed to corroborate these findings in the general population.


Fc-dependent IgG-mediated suppression of the antibody response: fact or artefact?
IgG antibodies have been shown to suppress the antibody response to all epitopes of their specific antigen as well as those the IgG do not bind to, so-called 'non-epitope-specific suppression'. The present study was undertaken to clarify whether there is a true IgG-mediated Fc-dependent suppression of the antibody response. This question is of fundamental importance to the understanding of the mechanism behind this phenomenon. It is demonstrated that F(ab')2 fragments of a monoclonal TNP (trinitrophenyl)-specific IgG2a antibody are unable to suppress the murine in vitro non-epitope-specific plaque-forming cell response against SRBC (sheep erythrocytes) when SRBC-TNP is used as antigen. The same monoclonal IgG antibody, when administered in intact form, is able to induce up to 98% suppression of the SRBC-specific antibody response. The lack of suppression is not due to mitogenic effects of pepsin in the F(ab')2 fractions or increased breakdown of F(ab')2 fragments, as compared with intact antibody, in the cultures. These data clearly demonstrate that there is indeed a highly efficient, Fc-dependent, non-epitope-specific suppressive mechanism mediated by IgG antibodies and support a hypothesis involving binding of the antigen-antibody complexes to Fc receptors as a step in the effector mechanism.


Computerized adaptive testing: the capitalization on chance problem.
This paper describes several simulation studies that examine the effects of capitalization on chance in the selection of items and the ability estimation in CAT, employing the 3-parameter logistic model. In order to generate different estimation errors for the item parameters, the calibration sample size was manipulated (N = 500, 1000 and 2000 subjects) as was the ratio of item bank size to test length (banks of 197 and 788 items, test lengths of 20 and 40 items), both in a CAT and in a random test. Results show that capitalization on chance is particularly serious in CAT, as revealed by the large positive bias found in the small sample calibration conditions. For broad ranges of theta, the overestimation of the precision (asymptotic Se) reaches levels of 40%, something that does not occur with the RMSE (theta). The problem is greater as the item bank size to test length ratio increases. Potential solutions were tested in a second study, where two exposure control methods were incorporated into the item selection algorithm. Some alternative solutions are discussed.


Effects of mandatory continuing education related to infection control on the infection control practices of dental hygienists.
The infection control (IC) practices of dental hygienists, who were practicing in states with and without mandatory continuing education (CE) related to IC, were compared using a nationwide survey approach. The percentages of respondents from states with mandated IC CE, who reported compliance with 8 practices described in the current IC guidelines and acquiring more CE units and taking more recent IC CE, were greater (P < .005) than the percentages of respondents from states without the requirement.


Analytical ultracentrifugation: still the gold standard that offers multiple solutions.
Understanding the nature of macromolecules and their interactions in solution underpins many fields, including biology, chemistry and materials science. The 24th International Analytical Ultracentrifugation Workshop and Symposium (AUC2019, held in Christchurch, New Zealand, August 2019), brought together 77 international delegates to highlight recent developments in the field. There was a focus on analytical ultracentrifugation, although we recognise that this is but one of the key methods in the biophysicist's toolkit. Many of the presentations showcased the versatility of analytical ultracentrifugation and how such experiments are integrated with other solution techniques, such as small-angle X-ray scattering, cryo-electron microscopy, isothermal titration calorimetry and more. This special issue emphasises a wide range of themes covered in the meeting, including carbohydrate chemistry, protein chemistry, polymer science, and macromolecular interactions.


Dengue virus requires the CC-chemokine receptor CCR5 for replication and infection development.
Dengue is a mosquito-borne disease that affects millions of people worldwide yearly. Currently, there is no vaccine or specific treatment available. Further investigation on dengue pathogenesis is required to better understand the disease and to identify potential therapeutic targets. The chemokine system has been implicated in dengue pathogenesis, although the specific role of chemokines and their receptors remains elusive. Here we describe the role of the CC-chemokine receptor CCR5 in Dengue virus (DENV-2) infection. In vitro experiments showed that CCR5 is a host factor required for DENV-2 replication in human and mouse macrophages. DENV-2 infection induces the expression of CCR5 ligands. Incubation with an antagonist prevents CCR5 activation and reduces DENV-2 positive-stranded (+) RNA inside macrophages. Using an immunocompetent mouse model of DENV-2 infection we found that CCR5(-/-) mice were resistant to lethal infection, presenting at least 100-fold reduction of viral load in target organs and significant reduction in disease severity. This phenotype was reproduced in wild-type mice treated with CCR5-blocking compounds. Therefore, CCR5 is a host factor required for DENV-2 replication and disease development. Targeting CCR5 might represent a therapeutic strategy for dengue fever. These data bring new insights on the association between viral infections and the chemokine receptor CCR5.


BRAF V600E mutation in Juvenile Xanthogranuloma family neoplasms of the central nervous system (CNS-JXG): a revised diagnostic algorithm to include pediatric Erdheim-Chester disease.
The family of juvenile xanthogranuloma family neoplasms (JXG) with ERK-pathway mutations are now classified within the "L" (Langerhans) group, which includes Langerhans cell histiocytosis (LCH) and Erdheim Chester disease (ECD). Although the BRAF V600E mutation constitutes the majority of molecular alterations in ECD and LCH, only three reported JXG neoplasms, all in male pediatric patients with localized central nervous system (CNS) involvement, are known to harbor the BRAF mutation. This retrospective case series seeks to redefine the clinicopathologic spectrum of pediatric CNS-JXG family neoplasms in the post-BRAF era, with a revised diagnostic algorithm to include pediatric ECD. Twenty-two CNS-JXG family lesions were retrieved from consult files with 64% (n = 14) having informative BRAF V600E mutational testing (molecular and/or VE1 immunohistochemistry). Of these, 71% (n = 10) were pediatric cases (≤18 years) and half (n = 5) harbored the BRAF V600E mutation. As compared to the BRAF wild-type cohort (WT), the BRAF V600E cohort had a similar mean age at diagnosis [BRAF V600E: 7 years (3-12 y), vs. WT: 7.6 years (1-18 y)] but demonstrated a stronger male/female ratio (BRAF V600E: 4 vs WT: 0.67), and had both more multifocal CNS disease ( BRAFV600E: 80% vs WT: 20%) and systemic disease (BRAF V600E: 40% vs WT: none). Radiographic features of CNS-JXG varied but typically included enhancing CNS mass lesion(s) with associated white matter changes in a subset of BRAF V600E neoplasms. After clinical-radiographic correlation, pediatric ECD was diagnosed in the BRAF V600E cohort. Treatment options varied, including surgical resection, chemotherapy, and targeted therapy with BRAF-inhibitor dabrafenib in one mutated case. BRAF V600E CNS-JXG neoplasms appear associated with male gender and aggressive disease presentation including pediatric ECD. We propose a revised diagnostic algorithm for CNS-JXG that includes an initial morphologic diagnosis with a final integrated diagnosis after clinical-radiographic and molecular correlation, in order to identify cases of pediatric ECD. Future studies with long-term follow-up are required to determine if pediatric BRAF V600E positive CNS-JXG neoplasms are a distinct entity in the L-group histiocytosis category or represent an expanded pediatric spectrum of ECD.


An analysis of cyclin D1, cytokeratin 5/6 and cytokeratin 8/18 expression in breast papillomas and papillary carcinomas.
To evaluate the expression levels of cyclin D1 in breast papillomas and papillary carcinomas, and to analyze the types of cells that co-express cyclin D1 with cytokeratin 5/6 (CK 5/6) or with cytokeratin 8/18(CK 8/18). Fifty-nine cases of papillary lesions including 36 papillomas and 23 intracystic papillary carcinomas were examined. Cyclin D1, CK 5/6 and CK 8/18 expression levels were evaluated by double immunostaining. Cyclin D1 is highly expressed in papillary carcinomas (27.54% ± 15.43%) compared with papillomas (8.81% ± 8.41%, p < 0.01). Cyclin D1 is predominantly expressed in cytokeratin 8/18- expressing cells, rather than in cytokeratin 5/6-expressing cells, regardless of the type of lesion. In papillomas, cyclin D1 exhibited a mean 11.42% (11.42% ± 10.17%) co-expression rate with cytokeratin 8/18 compared with a mean 2.50% (2.50% ± 3.24%) co-expression rate with cytokeratin 5/6 (p < 0.01). In papillary carcinomas, cyclin D1 exhibited a mean 34.74% (34.74% ± 16.32%) co-expression rate with cytokeratin 8/18 compared with a co-expression rate of 0.70% (0.70% ± 0.93%) with cytokeratin 5/6 (p < 0.01). The increase in cyclin D1 suggests an association of cyclin D1 staining with papillary carcinomas. Although cyclin D1 is an effective marker for the differential diagnosis of other papillary lesions, it cannot be used to distinguish between papilloma and papillary carcinoma lesions because its expression occurs in both lesions. Our results show that cyclin D1 and CK 5/6 staining could be used in concert to distinguish between the diagnosis of papilloma (cyclin D1 < 4.20%, CK 5/6 positive) or papillary carcinoma (cyclin D1 > 37.00%, CK 5/6 negative). In addition, our data suggest that cyclin D1 is expressed only in the cancer stem or progenitor cells that co-immunostained with CK 8/18 in papillary carcinomas, and predominantly with CK 8/18 in the papillomas. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/7299340558756848.


Results and insights after 413 TAVI procedures performed by cardiac surgeons on their own.
Current evidence on transcatheter aortic valve implantation (TAVI) has been generated exclusively by cardiology studies and no operative data from cardiac surgeons are available. Here, we describe the development of our TAVI programme and report the results of transfemoral (TF) TAVI done by cardiac surgeons on their own. This study included all the TAVI procedures on native valve performed at Cardiac Surgery Unit, Ospedali Riuniti di Ancona, during the period October 2018 to July 2022. Relevant prospectively collected preoperative, intraprocedural and postoperative data were retrieved from the Institutional database. A total of 413 patients were included in the study. Mean patients' age was 82 years and among them 44% (180/413) were male. STS score was 3.1% (2.2-4.4). Eighty patients underwent transapical TAVI and 333 patients had a TF approach. We progressively moved from transapical TAVI towards TF procedures that are now routinely performed on conscious sedation and using a fully percutaneous approach. After TF TAVI, 30-day mortality rate was 1%, cerebral stroke occurred in 2% of the cases, permanent pacemaker implantation was necessary in 23% of the patients and in 6% of the cases there was a moderate/severe degree of aortic regurgitation. There was no association between operators performing TAVI and 30-day mortality. The acquisition of catheter-based skills and an adequate training allowed cardiac surgeons to perform on their own awake and fully percutaneous TF TAVI with similar results when compared with major randomized clinical trials and registries' experiences.


Differentiation of reptilian neural crest cells in vitro.
An attempt was made to culture neural crest cells of the turtle embryo in vitro. Trunk neural tubes from the St. 9/10 embryos were explanted in culture dishes. The developmental potency of the turtle neural crest cells in vitro was shown to be essentially similar to that of avian neural crest cells, although they seem to be more sensitive to melanocyte-stimulating hormone (MSH) stimulation. We describe conditions under which explanted neural tube gives rise to neural crest cells that differentiate into neuronal cells and melanocytes. The potency of melanocyte differentiation was found to vary according to the concentration of fetal bovine serum (FBS, from 5 to 20%). Melanization of neural crest cells cultured in the medium containing FBS and alpha-MSH was more extensive than those cultured with FBS alone, combinations of FBS and chick embryo extract, or turtle embryo extract. These culture conditions seem to be useful for the study of the developmental potency of the neural crest cells as well as for investigating local environmental factors.


Persistence of indentation with bioabsorbable poly-L/D-lactide versus silicone sponge scleral buckling implants.
To measure the amount and duration of indentation depth achieved with biodegradable poly-L/D-lactide 96/4 (PLA96) and silicone sponge implants. Thirty rabbits underwent a scleral buckling procedure. A PLA96 buckling implant was used in 15 rabbits and a silicone sponge buckling implant was in 15 rabbits. A circumferential scleral buckling implant was sutured episclerally on the left eye of each rabbit, just temporal to the superior rectus muscle and 7 mm posterior to the limbus. Computed tomography was performed at 1 week, 3 months, and 5 months after surgery. The PLA96 buckling implant (implant diameter, 3-3.5 mm) used in this study created lower indentation than the silicone sponge implant (implant diameter, 4 mm). The indentation created by the PLA96 implant decreased over time compared with that created by the silicone implant. There were no complications related to either kind of implant. Both the silicone sponge implant and the PLA96 implant caused indentation that decreased in a comparable manner over the follow-up period (5 months).


SPME analysis of volatile compounds from unfermented olives subjected to thermal treatment.
The effect of different types of thermal treatment, designed to increase the product's shelf-life, on the volatile composition of "Campo Real" unfermented table olives, has been studied by headspace solid-phase microextraction (HS-SPME) and GC-MS analysis. Different SPME fibres were evaluated to determine their selectivity for a mixture of the main components of the different spices used in "Campo Real" olive dressing. Of the different fibres investigated, the polydimethylsiloxane/divinylbenzene (PDMS/DVB) fibre was selected for analysis of the olive brines, which contained nine main aroma components. The types of thermal treatment were sterilisation (121 degrees C, 15 min) and four pasteurisation conditions (60 degrees C or 80 degrees C each for 5 or 9 min). Pasteurisation did not lead to significant changes in the amounts of these nine volatile compounds; the 2-butanol signal was reduced by treatment at 80 degrees C. On the other hand, sterilisation of the brine resulted in an decrease in the signals from these compounds and the appearance of a new, high signal for benzaldehyde; the origin of this has not yet been determined. Results suggest that the selected pasteurisation conditions do not significantly modify the typical, and valued, aroma characteristics of "Campo Real".


Expression of p53 and flow cytometric DNA analysis of isolated neoplastic glands of the stomach: an application of the gland isolation method.
The expression of p53 was studied immunohistochemically in combination with the DNA ploidy pattern by gland isolation in 97 alcohol-fixed gastric lesions. A polyclonal antibody, CM-1, was applied to the paraffin-embedded sections in this study. Overexpression of the p53 protein was found in 73.2% of 41 well or moderately differentiated gastric carcinomas and 52.2% of 23 cases with poor differentiation (P < 0.05). Immunoreactivity of p53 was also detected in isolated cancerous glands. No p53 immunoreactivity was detected in benign gastric lesions including adenomas, hyperplastic polyps and regions of intestinal metaplasia. In addition, flow cytometric DNA analysis was performed on isolated glandular epithelium adjacent to the portions used for immunostaining. DNA aneuploidy (DA) was detected in 85.7% of the well or moderately differentiated carcinomas and 42.9% of those with poor differentiation (P < 0.05). There was a positive correlation between DA, p53 positivity and the presence of regional lymph node metastasis, but not with other clinicopathological variables. In spite of the limited applicability of this method to poorly differentiated gastric cancer, we found that immunostaining and flow cytometry in combination with the gland isolation method facilitates analysis of gastric carcinogenesis.


A DddA ortholog-based and transactivator-assisted nuclear and mitochondrial cytosine base editors with expanded target compatibility.
Bacterial double-stranded DNA (dsDNA) cytosine deaminase DddAtox-derived cytosine base editor (DdCBE) and its evolved variant, DddA11, guided by transcription-activator-like effector (TALE) proteins, enable mitochondrial DNA (mtDNA) editing at TC or HC (H = A, C, or T) sequence contexts, while it remains relatively unattainable for GC targets. Here, we identified a dsDNA deaminase originated from a Roseburia intestinalis interbacterial toxin (riDddAtox) and generated CRISPR-mediated nuclear DdCBEs (crDdCBEs) and mitochondrial CBEs (mitoCBEs) using split riDddAtox, which catalyzed C-to-T editing at both HC and GC targets in nuclear and mitochondrial genes. Moreover, transactivator (VP64, P65, or Rta) fusion to the tail of DddAtox- or riDddAtox-mediated crDdCBEs and mitoCBEs substantially improved nuclear and mtDNA editing efficiencies by up to 3.5- and 1.7-fold, respectively. We also used riDddAtox-based and Rta-assisted mitoCBE to efficiently stimulate disease-associated mtDNA mutations in cultured cells and in mouse embryos with conversion frequencies of up to 58% at non-TC targets.


Next of kin's experiences of registered nurses' leadership close to older adults in municipal home care in Sweden: an interview study.
Next of kin to older adults over 65 years in municipal home care are concerned whether their older adults' needs are being met. In municipal home care, the registered nurses' leadership is important and complex, entailing multi-artist skills involving the older adults and their next of kin. Yet, little is known about next of kin's experiences of registered nurses' leadership. Thus, the aim of this study was to explore next of kin's experiences of registered nurses' leadership close to older adults in municipal home care. Individual telephone interviews were conducted with next of kin (n = 11) of older adults from April to September 2020 in two municipalities in western Sweden. Data were analysed using qualitative content analysis. The results are presented with the theme, registered nurses do what they can, including two categories, interaction and competence, and the subcategories, relationship, communication, availability, responsibility, team leadership and cooperation. Registered nurses' leadership was experienced as a balancing act between their commitments and what they were able to achieve. Next of kin's experiences of registered nurses' leadership can contribute knowledge that will strengthen and prepare registered nurses for their leadership roles. This knowledge can support the development of policies for organisational preconditions that ensure quality and safe care to older adults in municipal home care.


Fungal biomarker monitoring and CT scans for early detection of invasive fungal disease in neutropenic hematological patients.
By using data from the CEDMIC trial (n = 413), we conducted a post-hoc analysis of the diagnostic value of biomarker monitoring and chest computed tomography (CT) scans for the early detection of invasive fungal disease (IFD) in neutropenic hematological patients. IFDs were defined in accordance with the EORTC/MSG definition with some modifications. Biomarkers such as Aspergillus galactomannan (GM) and (1→3)-β-D-glucan (βDG) were measured weekly. The positive predictive value (PPV) of GM and βDG in cases of high-risk treatment were 0.70 and 0.69, while those in low-risk treatment were 0.08 and 0, respectively. All of the positive biomarkers that were measured before the development of fever in low-risk treatment were false positives. The proportion of patients who had abnormal chest CT findings was 19% in persistent fever at 4-6 days, 57% at 7 days or later and 36% in recurrent fever. Sixty-nine percent of the patients who had abnormal findings at 7 days or later did not have abnormalities at 4-6 days. Afebrile screening of biomarkers in low-risk treatment is not useful. Chest CT should be reevaluated in persistent fever lasting for 7 days or longer even in patients who did not have abnormalities within 6 days.


An experimental steroid responsive model of ocular inflammation in rabbits using an SLT frequency doubled Q switched Nd:YAG laser.
To develop a minimally invasive rabbit model of postoperative anterior chamber (AC) inflammation using a commercially available frequency doubled Nd:YAG laser [intended for selective laser trabeculoplasty (SLT)]. Escalating laser energy was applied to the iris of male Dutch-belted rabbits and the subsequent inflammatory response was observed to determine the laser dose required to generate self-limiting inflammation of at least 3 days' duration. In subsequent experiments, 10 eyes of 10 male Dutch-belted rabbits underwent baseline slit lamp examination, intraocular pressure (IOP), and AC flare meter readings. Starting 1 day before laser application, 5 animals received topical 20 μL dexamethasone 1% to 1 eye 4 times daily for 5 days. Five control animals were treated with saline. Masked assessments of flare, cells, and IOP were made daily for 7 days. Histopathologic changes were assessed in enucleated eyes. Compared to controls, dexamethasone-treated rabbits had less postlaser AC flare on postoperative day (POD)2 (19±5 vs. 44±21photons/ms, P=0.03) and POD3 (16±9 vs. 33±11 photons/ms, P=0.03). In dexamethasone-treated rabbits, clinically graded flare (on POD1) and cells (on POD1 and 2) were lower than controls, but did not reach statistical significance. In the control group, IOP was significantly lower than the dexamethasone-treated group on POD2 (14.1±3.4 vs. 19.8±1.1 mmHg, P=0.03) and POD3 (14.2±2.2 vs. 19.0±2.2 mmHg, P=0.01). Histopathology showed pigment clumping and changes limited to anterior layers of the iris. Commercially available SLT laser can be used to create a minimally invasive, steroid-responsive animal model of anterior uveitis with the potential for use in the evaluation and comparison of drugs intended to treat AC inflammation.


Use of the Personality Assessment Inventory as an efficacious and cost-effective diagnostic tool for nonepileptic seizures.
Video electroencephalographic monitoring (VEEG) is considered the "gold standard" for making the differential diagnosis between epileptic seizures (ES) and nonepileptic seizures (NES), but is a costly, time-consuming procedure and not readily available in all communities. Of the various diagnostic techniques and measures that have been used, the Personality Assessment Inventory (PAI) has shown promise as an effective psychological screening tool to aid in the differential diagnosis of ES/NES. Using VEEG results as the outcome measure, this study examined the diagnostic effectiveness of the PAI in a group of adults with treatment-refractory seizures. Results indicated that, on psychological screening, patients with NES endorse significantly greater functional consequences of their seizure-like episodes than participants with ES. A "NES Indicator" score, calculated from the PAI Somatization subscales, provided a sensitivity of 84% and specificity of 73% for the diagnosis of NES versus ES. The PAI appears to be a useful screening tool prior to hospital admission for VEEG.