Multiple primer pairs for polymerase chain reaction (PCR) amplification of human parvovirus B19 DNA.
Human parvovirus B19 is the etiologic agent of erythema infectiosum and transient aplastic crisis in patients with hemolytic anemias and has been associated with fetal death, arthritis, and chronic anemia. Acute B19 infection is best diagnosed by detection of IgM antibodies, whereas the diagnosis of chronic infection often requires the sensitivity of PCR to demonstrate presence of virus over time. To improve our ability to detect B19 DNA by polymerase chain reaction (PCR), we evaluated 19 primers combined into 16 different primer pairs for their ability to detect temporally and geographically diverse B19 isolates. All 16 pairs reacted with all isolates tested but with different sensitivity. Sequence analysis showed few nucleotide changes compared with published sequences. These changes did not explain observed differences in sensitivity between primer pairs. The most sensitive primer pairs detected 350 to 3500 DNA copies after 35 cycles. A second amplification cycle with nested primers improved the sensitivity 100-fold. These 16 primer pairs provide the diagnostic virologist with multiple options for B19 PCR assays.


Presenile onset of spinocerebellar ataxia type 1 presenting with conspicuous psychiatric symptoms and widespread anti-expanded polyglutamine antibody- and fused in sarcoma antibody-immunopositive pathology.
A 50-year-old Japanese man showed slowly progressive gait disturbance and dysarthria. Neurological examination 5 years after onset revealed slow eye movement with nystagmus as well as limb and truncal ataxia. Magnetic resonance imaging showed atrophy of the cerebellum and brainstem. Because genetic examination revealed CAG repeat expansion of the ataxin-1 gene, the patient was diagnosed with spinocerebellar ataxia type 1. Ten years after onset, he showed psychiatric symptoms with cognitive impairment, and antipsychotic drugs were administered. As psychiatric symptoms gradually worsened, particularly with regard to resisting nursing care and shouting, the doses of the drugs were increased. Although the clinicopathologic findings were generally identical to previously reported spinocerebellar ataxia type 1 cases with the exception of the conspicuous psychiatric symptoms, there are two notable immunohistochemical findings. Firstly, numerous anti-expanded polyglutamine antibody-immunopositive neuronal inclusions were extensively observed, including in the cerebral cortex and limbic system, but not in the Purkinje cells. Secondly, anti-fused in sarcoma antibody-immunopositive intranuclear inclusions were extensively observed. We posit that the anti-expanded polyglutamine antibody-immunopositive neuronal inclusions and possibly the anti-fused in sarcoma antibody-immunopositive inclusions, particularly those in the neocortex and limbic system, may correspond to the psychiatric symptoms and cognitive impairment that were observed in the patient.


Machine learning approaches that use clinical, laboratory, and electrocardiogram data enhance the prediction of obstructive coronary artery disease.
Pretest probability (PTP) for assessing obstructive coronary artery disease (ObCAD) was updated to reduce overestimation. However, standard laboratory findings and electrocardiogram (ECG) raw data as first-line tests have not been evaluated for integration into the PTP estimation. Therefore, this study developed an ensemble model by adopting machine learning (ML) and deep learning (DL) algorithms with clinical, laboratory, and ECG data for the assessment of ObCAD. Data were extracted from the electronic medical records of patients with suspected ObCAD who underwent coronary angiography. With the ML algorithm, 27 clinical and laboratory data were included to identify ObCAD, whereas ECG waveform data were utilized with the DL algorithm. The ensemble method combined the clinical-laboratory and ECG models. We included 7907 patients between 2008 and 2020. The clinical and laboratory model showed an area under the curve (AUC) of 0.747; the ECG model had an AUC of 0.685. The ensemble model demonstrated the highest AUC of 0.767. The sensitivity, specificity, and F1 score of the ensemble model ObCAD were 0.761, 0.625, and 0.696, respectively. It demonstrated good performance and superior prediction over traditional PTP models. This may facilitate personalized decisions for ObCAD assessment and reduce PTP overestimation.


Two-stage culture procedure using thidiazuron for efficient micropropagation of Stevia rebaudiana, an anti-diabetic medicinal herb.
Stevia rebaudiana Bertoni, member of Asteraceae family, has bio-active compounds stevioside and rebaudioside which taste about 300 times sweeter than sucrose. It regulates blood sugar, prevents hypertension and tooth decay as well as used in treatment of skin disorders having high medicinal values, and hence there is a need for generating the plant on large scale. We have developed an efficient micropropagation protocol on half strength Murashige and Skoog (MS) media, using two-stage culture procedures. Varying concentrations of cytokinins, i.e., benzylaminopurine, kinetin and thidiazuron (TDZ) were supplemented in the nutrient media to observe their effects on shoot development. All the cytokinins promoted shoot formation, however, best response was observed in the TDZ (0.5 mg/l). The shoots from selected induction medium were sub-cultured on the multiplication media. The media containing 0.01 mg/l TDZ produced maximum number of shoot (11.00 ± 0.40) with longer shoots (7.17 ± 0.16) and highest number of leaves (61.00 ± 1.29). Rooting response was best observed in one-fourth strength on MS media supplemented with indole-3-butyric acid (1.0 mg/l) and activated charcoal (50 mg/l) with (11.00 ± 0.40) number of roots. The plantlets thus obtained were hardened and transferred to the pots with soil and sand mixture, where the survival rate was 80 % after 2 months. Quantitative analysis of stevioside content in leaves of in vivo mother plant and in vitro plantlets was carried out by high performance liquid chromatography. A remarkable increase in stevioside content was noticed in the in vitro-raised plants as compared to in vivo grown plants. The protocol reported here might be useful in genetic improvement and high stevioside production.


Computational investigation of the radical-mediated mechanism of formation of difluoro methyl oxindoles: Elucidation of the reaction selectivity and yields.
Oxindoles are an important class of heterocyclic alkaloids with demonstrated pharmacological activity at multiple biological targets. Preparation of new analogs through novel synthetic routes is therefore highly attractive. In this work, we report a computational study to investigate the synthesis of ethoxycarbonyldifluoromethylated oxindoles from N-arylmethacrylamides. The reaction tolerates a diverse range of acrylamides, shows yields ranging from approximately 38%-96%. We have applied density functional theory (DFT) to explore the reaction mechanism, kinetics and thermodynamics to gain further understanding. We demonstrate that a radical-based ring closure reaction is energetically more favorable than a heterolytic process, that the rate-determining step is the formation of the arylmethacrylamide radical, and that the product yields and selectivities are consistent with experiment. The results demonstrate that theoretical methods can prove useful to understand how such reaction and could be potentially employed to rapidly explore the reaction scope further.


Human papillomavirus subtypes in oral lesions compared to healthy oral mucosa.
Human papillomaviruses (HPV) are involved in the etiology of cervix cancer, but it is still unclear whether they play a role in related oral lesions. The presence of HPV in oral leukoplakia biopsies (n=50) and oral squamous carcinoma biopsies (n=50) was compared to normal oral mucosa swabs (n=50) for the purpose of indicating a possible etiological role for the virus. DNA was extracted from tissue biopsies and from mucosa swabs of control samples. Nested PCR was performed with primers targeting conserved sequences within the capsid gene L1. PCR products were sequenced to identify the HPV genotype. The results reveal a profile of low-risk HPV genotypes in oral leukoplakia similar to that in healthy controls, while HPV was less frequently observed in oral squamous carcinoma. HPV does not seem to represent an important causal factor for the development of oral leukoplakia or oral squamous carcinoma.


Reduced effective spin-orbital degeneracy and spin-orbital ordering in paramagnetic transition-metal oxides: Sr2IrO4 versus Sr2RhO4.
We discuss the notions of spin-orbital polarization and ordering in paramagnetic materials, and address their consequences in transition-metal oxides. Extending the combined density functional and dynamical mean field theory scheme to the case of materials with large spin-orbit interactions, we investigate the electronic excitations of the paramagnetic phases of Sr(2)IrO(4) and Sr(2)RhO(4). We show that the interplay of spin-orbit interactions, structural distortions and Coulomb interactions suppresses spin-orbital fluctuations. As a result, the room temperature phase of Sr(2)IrO(4) is a paramagnetic spin-orbitally ordered Mott insulator. In Sr(2)RhO(4), the effective spin-orbital degeneracy is reduced, but the material remains metallic, due to both, smaller spin-orbit and smaller Coulomb interactions. The corresponding spectra are in excellent agreement with photoemission data. Finally, we make predictions for the spectra of paramagnetic Sr(2)IrO(4).


Cystic fibrosis: cell culture classes in a Danish population.
The cystic fibrosis (CF) culture phenotype of dermal fibroblasts (metachromasia, metabolic cooperation) of a group of 131 Danish CF patients and their families were studied to determine the distribution of the two CF culture classes and their prognostic significance. Of these, 62.6% (82) were Class I, 31.3% (41) Class II and 61.1% (8) were proposed to be genetic compounds. The occurrence of Class II was twice that found in a group of patients from New York (13%) and Minnesota (18%). The prognosis for Class II CF patients was considered to be poorer as: (1) The initial diagnosis was made earlier in Class II than in Class I or the compounds (63% of Class II were diagnosed in the first year of life, as compared to 35% in Class I and 13% of the compounds). (2) Only 5% of the Class II patients survived over the age of 15 years, both being deceased at the end of the study in 1976, whereas 24% of Class I and 63% of the compounds were over 15 years at the end of the study. This research added further evidence for genetic heterogeneity within the clinical syndrome, cystic fibrosis.


Interstitial pneumonitis during rituximab-containing chemotherapy for primary central nervous system lymphomas: a case report and review of literature.
Primary central nervous system lymphoma(PCNSL) is a rare kind of non-Hodgkin lymphoma. Rituximab combined with high-dose methotrexate, cytarabine and dexamethasone (R-MAD regimen) were reported effective for PCNSL patients. Rituximab can cause several side effects, including fever, chills and rigors. In this case report, we demonstrate rituximab-induced interstitial pneumonitis in a PCNSL patient who has been treated with R-MAD regimen. The patient recovered after treatment and she remains complete remission after following consolidation chemotherapy. Here is no report of potential fatal complications of Rituximab like interstitial pneumonitis nowadays in PCNSL patients. As Rituximab is widely used, physicians should raise their awareness of this rare complication and detect RTX-ILD in early stage.


Mutational analysis of the Bradyrhizobium japonicum common nod genes and further nod box-linked genomic DNA regions.
By insertional and deletional marker replacement mutagenesis the common nod region of Bradyrhizobium japonicum was examined for the presence of additional, essential nodulation genes. An open reading frame located in the 800 bp large intergenic region between nodD1 and nodA did not appear to be essential for nodulation of soybean. Furthermore, a strain with a deletion of the nodI- and nodJ-like genes downstream of nodC had a Nod+ phenotype. A mutant with a 1.7 kb deletion immediately downstream of nodD1 considerably delayed the onset of nodulation. This region carried a second copy of nodD (nodD2). A nodD1-nodD2 double mutant had a similar phenotype to the nodD2 mutant. Using a 22-mer oligonucleotide probe partially identical to the nod box sequence, a total of six hybridizing regions were identified in B. japonicum genomic DNA and isolated from a cosmid library. Sequencing of the hybridizing regions revealed that at least three of them represented true nod box sequences whereas the others showed considerable deviations from the consensus sequence. One of the three nod box sequences was the one known to be associated with nodA, whereas the other two were located 60 to 70 kb away from nif cluster I. A deletion of one of these two sequences plus adjacent DNA material (mutant delta 308) led to a reduced nodulation on Vigna radiata but not on soybean. Thus, this region is probably involved in the determination of host specificity.


Synchronous teaching and asynchronous trauma: Exploring teacher trauma in the wake of Covid-19.
Following the onset of COVID-19, major subsections of our global society have shifted and mutated from the status quo of operation, especially in the field of education. A system that served far more social functions than just basic literacy and numeracy instruction, education has shifted out of the school buildings and into living rooms and spare bedrooms across the world. This research investigated the physical and psychological implications of traversing and functioning in this new virtual world that educators found themselves operating in. This study utilized narrative inquiry and survey data in order to gather mixed method data to gain a deeper understanding of the impact of COVID-19 and how educators processed and coped with the transition to distance learning. The findings illuminated narratives of trauma and managing stress in the face of the pandemic. Additional study may focus on replicating this research across multiple research locations.


Role of consolidative thoracic radiation in extensive-stage small-cell lung cancer with first-line chemoimmunotherapy: a retrospective study from a single cancer center.
To investigate the role of consolidative thoracic radiation (TRT) in extensive-stage small-cell lung cancer (ES-SCLC) receiving first-line chemo-immunotherapy followed by immunotherapy maintenance. Outcomes of patients without disease progression after first-line chemotherapy were retrospectively reviewed (January 2020 to December 2021). Based on TRT or not, patients were allocated to TRT group or non-TRT group. Progression-free survival (PFS), overall survival (OS) and local-recurrence free survival (LRFS) were calculated by the Kaplan-Meier method and compared by log-rank test. Of 100 patients, 47 received TRT and 53 non-TRT. The median follow-up was 20.3 months. The median PFS and OS in TRT were 9.1 months and 21.8 months, versus 8.8 months (p = 0.93) and 24.3 months (p = 0.63), respectively, in non-TRT. The median LRFS time in TRT was not reached, but significantly longer than 10.8 months in non-TRT (HR = 0.27, p < 0.01). Second-line chemotherapy significantly prolonged survival compared to that with chemo-free patients (mOS: 24.5 vs. 21.4 months, p = 0.026). The subgroup analysis showed a trend of patients with brain metastases benefit from TRT (21.8 versus 13.7 months, HR 0.61, p = 0.38) while liver metastases did not. Of 47 patients with TRT, only 10.6% of patients experienced grade 3 radiation-induced pneumonitis, while no grade 4 or 5 adverse events occurred. Consolidative TRT in the period of immunotherapy maintenance followed first-line chemo-immunotherapy did not prolong OS and PFS but associated with improved LRFS in ES-SCLC.


Parametric optimization of hexavalent chromium removal by electrocoagulation technology with vertical rotating cylindrical aluminum electrodes using Taguchi and ANN model.
This study aims to evaluate the performance of rotating aluminum electrodes in the electrocoagulation reactor for removing hexavalent chromium (Cr6+) from synthetic tannery wastewater. Taguchi and Artificial Neural Network (ANN) based models were developed to obtain the optimum condition for maximum Cr6+ removal. The optimum working condition obtained by Taguchi approach for the maximum Cr6+ removal (94%) was: Initial Cr6+ concentration (Cr6+ i) = 15 mg/L; Current Density (CD) = 14.25 mA/cm2; Intial pH = 5; Rotational Speed of Electrode (RSE) = 70 rpm. In contrast, the optimal condition for maximum Cr6+ ions removal (98.83%) obtained from the BR-ANN model was: Cr6+ i = 15 mg/L; CD = 14.36 mA/cm2; pHi = 5.2; RSE = 73 rpm. Compared to the Taguchi model, the BR-ANN model outperformed in terms of providing higher Cr6+ removal (+ 4.83%); reduced energy demand (-0.035 KWh/gm Cr6+ remove); lower error function value (χ2 = -7.9674 and RMSE = -3.5414); and highest R2 value (0.9991). The data for the conditions 91,007 < Re < 227,517 and Sc = 102.834 were found to fit the equation for the initial Cr6+ concentration of 15 mg/l; Sh = 3.143Re0.125 Sc0.33. The Cr6+ removal kinetics was best described by Pseudo 2nd Order model, as validated by high R2 and lower error functions value. The SEM and XRF analysis confirmed that Cr6+ was adsorbed and precipitated along with metal hydroxide sludge. The rotating electrode led to lower SEEC (10.25 kWh/m3), as well as maximum Cr6+ removal (98.83%), compared to EC process with stationary electrodes.


The susceptibility of Philadelphia chromosome positive cells to FAS-mediated apoptosis is not linked to the tyrosine kinase activity of BCR-ABL.
We investigated whether inhibition of the BCR-ABL tyrosine kinase by the CGP57418B compound would render chronic myeloid leukaemia (CML) cells susceptible to Fas (CD95, Apo-1)-mediated cell death. Only two (AR230 and SD1) out of 10 BCR-ABL positive cell lines were found to express the CD95 protein. No change in Fas expression was observed in any of the 10 cell lines after 48 h exposure to CGP57418B. AR230 cells were resistant and SD1 cells were partially resistant to Fas-mediated apoptosis induced by ligation of the Fas receptor to an anti-Fas IgM antibody. Pre-incubation with 1 microM CGP57418B did not change the susceptibility of these cell lines to Fas-mediated cell death. Similar results were observed in experiments with CD34+ cells from CML patients and from normal individuals. The data suggest that, in contrast to some cytotoxic drugs, the CGP57148B tyrosine kinase inhibitor utilizes a pathway other than the CD95 system in order to induce apoptosis in CML cells.


Thermal stability and activity regain of horseradish peroxidase in aqueous mixtures of imidazolium-based ionic liquids.
Thermal deactivation kinetics of horseradish peroxidase (HRP) were studied from 45 to 90 degrees C in phosphate buffer and 5-25% (v,w/v) 1-butyl-3-methylimidazolium tetrafluoroborate [BMIM][BF4] and 1-butyl-3-methylimidazolium chloride [BMIM][Cl]. HRP activity at 25 degrees C was not affected by the presence of ionic liquids up to 20% (v,w/v). Increasing the ionic liquids concentration up to 25% (v,w/v) changed the biphasic character of deactivation kinetics to an apparent single first-order step. The presence of 5-10% (v/v) [BMIM][BF4] significantly improved HRP thermal stability with lower activation energies for the deactivation second phase (83-87 kJ mol(-1)). After deactivation, enhanced activity regain of the enzyme, up to 70-80% of the initial activity, was found in 25% (v/v) [BMIM][BF4] and 10% (w/v) [BMIM][Cl] and correlated to prevalence of the deactivation first phase.


Characterization of a synaptiform transmission between a neuron and a glial cell in the leech central nervous system.
The cross-talk between neurons and glial cells is receiving increased attention because of its potential role in information processing in nervous systems. Stimulation of a single identifiable neuron, the neurosecretory Leydig interneuron in segmental ganglia of the leech Hirudo medicinalis, which modulates specific behaviors in the leech, evokes membrane hyperpolarization directly in the giant glial cell (Schmidt and Deitmer. Eur J Neurosci 11:3125-3133, 1999). We have studied the neuron-to-glia signal transmission in the voltage-clamped giant glial cell to determine whether this interaction exhibits properties of a chemical synapse. The glial response had a mean latency of 4.9 s and was dependent on the action potential frequency; the glial cell responded to as few as five Leydig neuron action potentials in 50% of the trials. The glial current was sustained for minutes during repetitive Leydig neuron activity without any sign of desensitization. The current was sensitive to tetraethylammonium, and its reversal potential of -78 mV shifted with the external K+ concentration. The glial response increased with the duration of the neuronal action potentials and was sensitive to the external Ca2+/Mg2+ concentration ratio. The results suggest that Leydig neuron activity leads to a Ca2+-dependent release of transmitter from the neuronal dendrites, evoking an K+ outward current in the giant glial cell, implying a synapse-like transmission between a neuron and a glial cell.


Mediation of subacute anthracycline cardiotoxicity in rabbits by cardiac histamine release.
We tested the hypothesis that cardiac histamine release mediates subacute doxorubicin (DXR) cardiotoxicity in rabbits. New Zealand white rabbits given DXR 20 mg/kg i.v. over 30 min developed myocardial damage 24 h later that was similar to that observed in humans. In isolated heart preparations, DXR produced dose-related cardiac histamine release at DXR concentrations of 1, 5, and 25 micrograms/ml given as 1-min exposures. Prior exposure of isolated hearts to 10 microM cromolyn sodium completely prevented histamine release from 5 micrograms/ml DXR. Pretreatment of animals with cromolyn produced significant protection against DXR-mediated subacute cardiotoxicity. We conclude that the release of cardiac histamine may be involved in the pathogenesis of anthracycline cardiotoxicity.


Carcinoid of the cervix: natural history and implications for therapy.
Carcinoid tumors rarely occur in the cervix. We present a patient with FIGO stage IB disease who developed brain metastases 4 years after initial diagnosis and treatment. The clinical course of a collected series of patients with carcinoid of the cervix is reviewed. Even patients with low stage lesions frequently die of disseminated disease. Adjuvant chemotherapy may have a role in the treatment of such patients.


The picture superiority effect in a cross-modality recognition task.
Words and pictures were studied and recognition tests given in which each studied object was to be recognized in both word and picture format. The main dependent variable was the latency of the recognition decision. The purpose was to investigate the effects of study modality (word or picture), of congruence between study and test modalities, and of priming resulting from repeated testing. Experiments 1 and 2 used the same basic design, but the latter also varied retention interval. Experiment 3 added a manipulation of instructions to name studied objects, and Experiment 4 deviated from the others by presenting both picture and word referring to the same object together for study. The results showed that congruence between study and test modalities consistently facilitated recognition. Furthermore, items studied as pictures were more rapidly recognized than were items studied as words. With repeated testing, the second instance was affected by its predecessor, but the facilitating effect of picture-to-word priming exceeded that of word-to-picture priming. The finds suggest a two- stage recognition process, in which the first is based on perceptual familiarity and the second uses semantic links for a retrieval search. Common-code theories that grant privileged access to the semantic code for pictures or, alternatively, dual-code theories that assume mnemonic superiority for the image code are supported by the findings. Explanations of the picture superiority effect as resulting from dual encoding of pictures are not supported by the data.


Childhood rheumatic diseases: bites not only the joint, but also the heart.
Cardiovascular involvement in juvenile rheumatic diseases is the primary manifestation in paediatric vasculitis and a major organ manifestation in paediatric connective tissue diseases. Though coronary vasculitis is the prototypical manifestation of Kawasaki disease, it can also be seen in patients with polyarteritis nodosa. Pericarditis is the most common manifestation seen in juvenile rheumatic diseases like systemic onset JIA, and lupus. Cardiac tamponade, valvular insufficiency, aortic root dilatation and arrhythmias are seen rarely. Cardiac involvement is often recognized late in children. The development of cardiac disease in juvenile systemic sclerosis is associated with a poor outcome. In long term, childhood onset of rheumatic diseases predisposes to diastolic dysfunction and premature atherosclerosis during adulthood. Key Points • Pericarditis is the most common cardiac manifestation in SLE and can lead to tamponade. • Conduction defects are common in juvenile mixed connective tissue disease and systemic sclerosis. • Pulmonary hypertension is a significant contributor to mortality in juvenile systemic sclerosis. • In Kawasaki disease, early treatment can reduce risk of coronary artery aneurysms.


Mentor inflatable penile prosthesis.
Technical complications requiring frequent surgical revision of the most commonly used penile component prosthesis have led to the development of a new inflatable device that has been implanted in 202 patients. This hydraulic device provides excellent functional qualities; no connector separations, aneurysms, or ruptures have been encountered in these patients. Reoperations became necessary in 8 per cent of these patients. Cosmetic and functional results are good, and patient acceptance has been excellent.


Reduction of platelet phospholipase C-delta1 activity in Alzheimer's disease associated with a specific apolipoprotein E genotype (epsilon3/epsilon3).
The epsilon4 allele of apolipoprotein E (apo E) is increased among patients with sporadic or familial Alzheimer's disease (AD). We examined platelet phospholipase C (PLC)-delta1 activity in AD patients either homozygous for apoepsilon3 or having at least one apo epsilon4 allele. We found that platelet PLC-delta1 activity is reduced from control levels in patients homozygous for apo epsilon3, but not changed in patients with an apo epilson4 allele. Reduced PLC-delta1 activity and apo epsilon3 may contribute to AD pathogenesis apart from the apo epsilon4 allele.


Relationship between lifespan and somatic mutation in D. melanogaster after treatment with chlorophyllin.
Sodium copper chlorophyllin (SCC) has a genetic damage inhibitory capacity due to its antioxidant action. For this reason, it was considered to investigate its role in the life span of Drosophila melanogaster and its relationship with the frequency of somatic mutation induced by gamma rays. Results indicated that SCC alone prolonged the lifespan only in females, but in combination with 20 Gy of gamma rays, the aging delay in both sexes was significant. In addition to confirming that the porphyrin reduces the frequency of mutation, the individuals with the highest mutation load are the individuals who die more quickly, and once they are eliminated, the survivor individuals treated with 20 Gy or with SCC + 20 Gy, died at the same rate. The results together indicate that SCC not only inhibits induced genetic damage, but it also has beneficial effects that probably cause an aging delay of the treated population that need to be investigated.


Single-Molecule Imaging to Characterize the Transport Mechanism of the Nuclear Pore Complex.
In the eukaryotic cell, a large macromolecular channel, known as the Nuclear Pore Complex (NPC), mediates all molecular transport between the nucleus and cytoplasm. In recent years, single-molecule fluorescence (SMF) imaging has emerged as a powerful tool to study the molecular mechanism of transport through the NPC. More recently, techniques such as single-molecule localization microscopy (SMLM) have enabled the spatial and temporal distribution of cargos, transport receptors and even structural components of the NPC to be determined with nanometre accuracy. In this protocol, we describe a method to study the position and/or motion of individual molecules transiting through the NPC with high spatial and temporal precision.


Determinants of help-seeking and system related components of delay in the treatment of first-episode psychosis.
Knowledge about factors that influence different components of duration of untreated psychosis (DUP) is important for designing interventions to reduce DUP. We tested associations between help-seeking and referral components of DUP (DUP-H and DUP-R, respectively) and the following predictor variables: age, gender, ethnicity, living arrangement, pre-morbid adjustment, age at onset of psychosis, diagnosis, level of symptoms, type of first and total number of mental health contacts prior to and after the onset of psychosis in a sample of 98 first-episode psychosis patients (FEP). Longer DUP-HS was significantly associated with earlier age at onset, diagnosis of schizophrenia spectrum psychosis and poor pre-morbid adjustment during adolescence. Longer DUP-R was associated with earlier age at onset and first help-seeking contact having been made with a non-medical professional. Relatively non-malleable patient characteristics are likely to influence delay in help-seeking while more malleable systemic characteristics influence delay associated with referral for specialized treatment.


Urgent laparoscopic repair of acutely symptomatic PEH is safe and effective.
Acute incarceration of paraesophageal hernias (PEHs) requiring urgent or emergent surgery is rare. Patients are often elderly with significant comorbidities and have historically been treated with open abdominal or thoracic incisions. Our study was designed to evaluate the feasibility, safety, and efficacy of laparoscopic paraesophageal hernia repair (LPEHR) in patients with PEH and acute gastric volvulus. We reviewed our prospectively maintained database and identified 269 patients who underwent an initial LPEHR between January 2003 and January 2012. Patients were divided into group A (acute), group B (age- and comorbidity-matched 1:3), and group C (all elective repairs). Group A included those admitted with acute symptoms related to PEH and underwent urgent repair. Patient age, Charlson score, operative time, length of stay (LOS), morbidity, mortality, and recurrence rates were compared. Patients who underwent urgent LPEHR had a higher perioperative morbidity rate than the elective and matched groups. The overall mortality rate was low and no statistical difference was found between groups A, B, and C. LOS in group A was longer than groups B and C. The need for ICU admission was also higher in group A. There was no statistical difference in recurrence rates. Historically, patients presenting with acute symptoms related to PEH have required open repair, which is associated with significant morbidity and mortality. The acute group was older and sicker than our elective LPEHR patients and had more adverse events resulting in a longer LOS, even when compared with comorbidity-matched elective patients. However, the LOS remained shorter than that reported for open repair and there was no mortality. The recurrence rates in all groups were low and comparable to elective repairs.


Correction to: Factors driving adaptive radiation in plants of oceanic islands: a case study from the Juan Fernández Archipelago.
The article Factors driving adaptive radiation in plants of oceanic islands: a case study from the Juan Fernández Archipelago, written by Koji Takayama, Daniel J. Crawford, Patricio López‑Sepúlveda, Josef Greimler, Tod F. Stuessy was originally published electronically on the publisher's internet portal (currently SpringerLink) on 13 March 2018 without open access.


Hematopoietic stem cell transplantation in patients with myelodysplastic syndrome.
Myelodysplastic syndrome (MDS) is a stem cell disorder, and hematopoietic stem cell transplantation is currently the only therapeutic modality that is potentially curative. Among patients with less advanced MDS (<5% marrow blasts), 3-year survivals of 65-70% are achievable with HLA-identical related and HLA-matched unrelated donors. The overall probability of disease recurrence in these patients is <5%. Among patients with advanced disease (>/=5% marrow blasts), about 35-45% and 25-30%, respectively, are surviving in remission after transplantation from a related or from an unrelated donor; the incidence of post-transplant relapse is 10-35%. The criteria proposed by the International Prognostic Scoring System (IPSS) derived from non-transplanted patients, also predict survival following transplantation. The development of new conditioning regimens has permitted successful hematopoietic stem cell transplants even in patients more than 60 years of age. Improved survival with transplants from unrelated volunteer donors may, in part, reflect selection of donors on the basis of high resolution (allele-level) HLA typing. Autologous stem cell transplantation may be beneficial for selected patients who have obtained a complete remission with conventional chemotherapy. Treatment-related morbidity and mortality, in particular after allogeneic transplantation, remain challenges that need to be addressed with innovative approaches.


Iris synechia after laser goniopuncture in a patient having deep sclerectomy with a collagen implant.
Neodymium:YAG (Nd:YAG) goniopuncture is an efficient and safe treatment for low filtration through the trabeculo-Descemet's membrane after deep sclerectomy with a collagen implant (DSCI). The only reported complication of this procedure is choroidal detachment. However, we found an iris synechia in a patient whose intraocular pressure (IOP) was elevated again 1 month after Nd:YAG goniopuncture. Synechiolysis and peripheral iridectomy with Nd:YAG and argon lasers effectively removed the iris synechia, and IOP immediately dropped to the normal range. We believe that iris synechia is a potential complication that may cause elevated IOP after laser goniopuncture in patients having DSCI.


Comparison of stimulants and hallucinogens on shuttle avoidance in rats.
Rats were trained to a high level of performance of a conditioned avoidance response in a shuttlebox to test effects of several classical stimulants in comparison to a variety of hallucinogens. A previously-reported biphasic pattern of effects of mescaline on shuttle avoidance was replicated and extended to 12 other hallucinogens of both phenylethylamine and indolealkylamine classes. Response patterns of hallucinogens could be differentiated from 3 stimulants and from a methoxyamphetamine compound that lacks hallucinogenic activity.


Biologics for childhood systemic vasculitis.
Recent advances have allowed better understanding of vasculitis pathogenesis and led to more targeted therapies. Two pivotal randomized controlled trials, RITUXVAS and rituximab in ANCA-associated vasculitis (RAVE), provide high-quality evidence demonstrating rituximab (RTX) is efficacious in inducing remission in adult ANCA-associated vasculitis (AAV) patients compared with cyclophosphamide (CYC). RAVE also demonstrated superiority of RTX to oral CYC for induction of remission in relapsing disease. Disappointingly, the RTX regimen was not associated with reduction in early serious adverse events. At least nine randomized trials are in progress, aiming to further delineate optimal dosing and duration of RTX therapy in AAV. In particular, the 6-month interim results of the PEPRS trial provide encouraging data specific to children. Due to special concerns related to growth, preservation of fertility, and potential for high cumulative medication doses, children with AAV should be considered as candidates for RTX even as a first-line remission induction therapy. Two randomized clinical trials have defined the role of infliximab in Kawasaki disease (KD), which appears to be as an alternative to a second infusion of intravenous immunoglobulin (IVIG) for treatment-resistant disease. Support for other biologics in the treatment of AAV or for biologics in the treatment of other vasculidities is largely lacking due to either unimpressive trial results or lack of trials. Except for the KD trials and the PEPRS, trials enrolling children remain scant. This review touches on the key trials and case series with biologics in the treatment of vasculitis that have influenced practice and shaped current thinking.


Gelation of the amoebocyte lysate of Tachypleus tridentatus by cell wall digest of several gram-positive bacteria and synthetic peptidoglycan subunits of natural and unnatural configurations.
Water-soluble adjuvant-active fractions prepared by digestion of the cell walls from several gram-positive bacteria with an endo-N-acetylmuramidase (Mutanolysin) were shown to gelate the amoebocyte lysate of Tachypleus tridentatus, the Japanese horseshoe crab. This finding was further extended by examination of the clotting activities on the amoebocyte lysate of a synthetic N-acetylmuramyl-L-alanyl-D-isoglutamine and related compounds, either adjuvant-active or -inactive, demonstrating that there are in general close correlations between the abilities to initiate lysate gelation, immunoadjuvancies and pyrogenicities, although there are some exceptions.


Caloric intake in a group of peritoneal dialysis patients.
Dialysis patients as a group appear to have a caloric intake less than the recommended values; because their energy expenditure is not different than that of healthy adults, they should be in negative caloric balance and lose body mass progressively. We retrospectively analyzed our data in a group of peritoneal dialysis patients who had dietary evaluations and kinetic measurements performed two consecutive times over a period of 6 months. Body weight, lean body mass (LBM), fat mass, and anthropometric parameters remained stable over this period of time, suggesting that these patients are not in negative energy balance. When their daily caloric intake was normalized to a new ideal body weight derived from the LBM calculated from creatinine kinetics, the values were within normal limits, suggesting that these patients were in zero energy balance; hence, a stable body mass was expected.


Reliability and number of trials of Y Balance Test in adolescent athletes.
The Star Excursion Balance Test (SEBT) is commonly used to evaluate dynamic equilibrium. The Y Balance Test (Y-BT) is a shortened version of the SEBT where a Y- Balance Kit is commonly used. To date, research concerning the protocol and reliability of the SEBT and Y-BT has been conducted only for adults. The aim of the study was to assess the protocol (the necessary number of trials to stabilize the results) and reliability of the Y-BT in adolescent athletes. One-way repeated-measures analysis of variance (ANOVA) and reliability study. The sample of 38 athletes (mean age: 15.6 years) was selected from a football club. A Y-Balance test kit was applied for the evaluation of dynamic balance. The analysis used the values normalized to the relative length of the lower limbs. After six attempts, three consecutive ones achieved stability for all directions and both extremities (p > 0.05). The intraclass correlation coefficient (ICC3,1), standard error of measurement and minimal detectable change values for the three attempts ranged from 0.57 to 0.82, from 3 to less than 6% and from 7.68 to 13.7%, respectively. In the study of adolescent dynamic equilibrium using the Y-BT, it is recommended to perform nine attempts (including six trial attempts and three measurements). In order to increase reliability it is recommended that the average of the three measured attempts is analysed.


Analysis of immunomodulating nitric oxide, iNOS and cytokines mRNA in mouse macrophages induced by microcystin-LR.
Microcystins (MCs) are the toxic molecules produced by common cyanobacterium in freshwater blooms. Their toxicities have brought severe health issues to livestock and human being. Microcystin-LR (MC-LR) is one of the most toxic MCs. This paper presents the profile of the immunomodulation of MC-LR to BALB/c mice peritoneal macrophages. Macrophages were stimulated with 100microg/l lipopolysaccharide (LPS) and MC-LR at dose of 1, 10, 100, 1000nmol/l, respectively, for 24h. Nitric oxide (NO) production in cell culture supernatants was quantified by using Griess reagent method. Total RNA was extracted from incubated macrophages then the mRNA abundance of induced nitric oxide synthase (iNOS), IL-1beta, TNF-alpha, GM-CSF, IFN-gamma was monitored by using reverse-transcriptional polymerase chain reaction (RT-PCR). The results demonstrated that NO production, mRNA levels of iNOS, IL-1beta, TNF-alpha were down regulated by MC-LR dose-dependently and mRNA levels of GM-CSF and IFN-gamma were also decreased but in dose-independent manner. Our results illustrated the involvement of NO production, iNOS and some cytokines in mice immune system in microcystin shock.


ST segment changes in exercise body surface mapping after myocardial infarction in patients with isolated left anterior descending coronary artery disease.
To investigate the clinical significance of exercise-induced ST segment elevation and ST segment depression after myocardial infarction (MI), we performed 87-lead ECG mapping after previous anterior infarction in 24 patients with isolated left anterior descending coronary artery disease before and 1.5 minutes after treadmill exercise. Thirteen patients showed ST segment elevation only, seven patients showed both ST segment elevation and depression, and four patients showed ST segment depression only. ST segment elevation most frequently occurred in the left anterior chest leads corresponding to the QS area, and ST segment depression developed in the left lower chest and left lower back leads. There was good correlation between the number of lead points showing ST segment elevation (nSTe) after exercise and the number of lead points showing QS waves (nQS) before exercise (r = 0.65). nSTe was also correlated with the asynergy index (r = 0.43). These findings suggest that ST segment elevation is mainly the result of aggravation of wall motion abnormalities of the infarcted myocardium. Body surface distribution of ST segment depression was similar to that in effort angina pectoris without MI. We conclude that exercise-induced ST segment depression in MI mainly reflects the ischemia of the surviving myocardium of small infarcts or the peripheral area of large infarcts.


Phosphoenolpyruvate carboxykinase ferroactivator 1. Mechanism of action and identity with glutathione peroxidase.
A cytosolic protein factor (ferroactivator) facilitates the activation of phosphoenolpyruvate carboxykinase by ferrous ions (Bentle, L. A., and Lardy, H. A. (1977) J. Biol. Chem. 252, 1431-1440). We have extended our studies on the interaction of Fe2+ with this enzyme to establish the conditions under which it is an activator or an inhibitor. Preincubation of phosphoenolpyruvate carboxykinase with Fe2+ and dithiothreitol resulted in irreversible loss of enzyme activity within minutes of Fe2+ addition. This was attributed to an active oxygen species produced by aerobic oxidation of the divalent metal ion in the presence of dithiothreitol as suggested by lack of inhibition in preincubation experiments with Fe2+ under mildly acidic pH; ferroactivation by many H2O2 scavenging enzymes; and lack of inhibition on preincubation under anaerobic conditions. We conclude that Fe2+ per se can activate phosphoenolpyruvate carboxykinase and that ferroactivator protein helps to overcome the deleterious effects of aerobic oxidation. Mechanistic details of ferroactivation and a comparison of the known properties of ferroactivator indicated the similarity of this protein with rat liver glutathione peroxidase. The identity of ferroactivator as glutathione peroxidase was confirmed by the demonstration of catalytic activity, selenium content, and immunological cross-reactivity.


NK cell alloreactivity and allogeneic hematopoietic stem cell transplantation.
As only 60% of leukaemia patients find a matched donor, the Perugia Bone Marrow Transplant Centre developed transplantation from HLA haplotype-mismatched family donors to provide a cure for more patients [F. Aversa, A. Tabilio, A. Terenzi, et al., Successful engraftment of T-cell-depleted haploidentical "three-loci" incompatible transplants in leukemia patients by addition of recombinant human granulocyte colony-stimulating factor-mobilized peripheral blood progenitor cells to bone marrow inoculum, Blood 84 (1994) 3948-3955] [F. Aversa, A. Tabilio, A. Velardi, et al., Treatment of high-risk acute leukemia with T-cell-depleted stem cells from related donors with one fully mismatched HLA haplotype, N. Engl. J. Med. 339 (1998) 1186-1193] [F. Aversa, A. Terenzi, A. Tabilio, et al., Full haplotype-mismatched hematopoietic stem-cell transplantation: a phase II study in patients with acute leukemia at high risk of relapse, J. Clin. Oncol. 23 (2005) 3447-3454]. HLA-mismatches trigger donor vs. recipient NK cell alloreactivity which improves engraftment, protects from GvHD and reduces relapse in AML patients [L. Ruggeri, M. Capanni, E. Urbani, et al., Effectiveness of donor natural killer cell alloreactivity in mismatched hematopoietic transplants, Science 295 (2002) 2097-2100], [L. Ruggeri, A. Mancusi, M. Capanni, E. Urbani, A. Carotti, T. Aloisi, M. Stern, D. Pende, K. Perruccio, E. Burchielli, F. Topini, E. Bianchi, F. Aversa, M.F. Martelli, A. Velardi, Donor natural killer cell allorecognition of missing self in haploidentical hematopoietic transplantation for acute myeloid leukemia: challenging its predictive value, Blood, in press]. We are using murine transplant models to determine whether NK cell alloreactivity can be exploited to reduce transplant-related mortality (TRM) which remains a major issue. Data from these on-going studies show pre-transplant infusion of alloreactive NK cells: (1) ablates AML cells, (2) kills recipient T cells, permitting a reduced toxicity conditioning regimen, and (3) ablates the recipient dendritic cells (DCs) which trigger GvHD, thus protecting from GvHD while permitting a higher T cell content in the graft. We are designing a clinical haploidentical transplant trial using alloreactive NK cells in the conditioning regimen, with the aim of reducing TRM and improving outcomes and overall survival.


Generation of hairpin-based RNAi vectors for biological and therapeutic application.
RNA interference (RNAi) is a natural process of gene silencing mediated by small RNAs. Shortly after the discovery of the RNAi mechanism, scientists devised various methods of delivering small interfering RNAs (siRNAs) capable of co-opting the endogenous RNAi machinery and suppressing target gene expression based on sequence complementarity. RNAi has since become a powerful tool to study gene function and is being investigated as a potential therapeutic approach to treat a vast array of human diseases (e.g., cancer, viral infections, and dominant genetic disorders). Among the available RNAi vectors are hairpin-based expression platforms (short-hairpin RNAs and artificial microRNAs) designed to mimic endogenously expressed inhibitory RNAs. These RNAi vectors are capable of achieving long-term potent gene silencing in vitro and in vivo. Here, we describe methods to design and generate these hairpin-based vectors and briefly review considerations for downstream applications.


[Kartagener's syndrome found incidentally during assessment of infertile couple: a case study].
Kartagener's syndrome is a rare primitive ciliary dyskinesia (DCP) characterized by a clinical triad: sinusitis, bronchiectasis and complete or incomplete situs inversus. It is a rare congenital autosomal recessive disease. We report a case of Kartagener syndrome in an infertile couple with akinospermia detected using spermogram.


Value of FDG-PET/CT examinations in different cancers of children, focusing on lymphomas.
The aim of the study was to assess sensitivity and specificity of FDG-PET/CT in different forms of childhood cancer. We retrospectively evaluated the results dedicated of 162 FDG-PET/CT examinations of 86 children treated with: Hodgkin lymphoma (HL; n = 31), non-Hodgkin lymphoma (NHL; n = 30) and other high grade solid tumors (n = 25). Patients were admitted and treated in two departments of pediatric hematology and oncology in Hungary. FDG-PET/CT was performed for staging (n = 25) and for posttreatment evaluation (n = 137). Imaging was performed in three FDG-PET/CT Laboratories, using dedicated PET/CT scanners. False positive results were defined as resolution or absence of disease progression over at least 1 year on FDG-PET/CT scans without any intervention. In some cases histopathological evaluation of suspicious lesions was performed. Fals negative results were defined as negative FDG-PET/CT results in case of active malignancy. Positive predictive values (PPV) and negative predictive values (NPV) were calculated. NPV was 100%. The highest PPV was observed in high grade solid tumors (81%), followed by HL (65%) and NHL (61%). There was a major difference of PPV in different histological types of HL (50% in HL of mixed-cellularity subtype, 90% in nodular sclerosing, and 100% in lymphocyte-rich and lymphocyte depleted HL). We treated one patient with nodular lymphocyte predominant HL, who had 5 false positive FDG-PET/CT results. PPV of T- and B-lineage NHL were similar (60% and 62%, respectively). We observed an interesting difference of PPV in different stages of HL and NHL. In HL PPV was higher in early than in advanced disease forms: 66% in stage II HL and 60% in stage III HL, whereas there was an inverse relationship between PPV and disease stages in NHL 0% in stage I and II patients, 67% in stage III and 100% in stage IV patients. PPV was lower in males (54%) than in females (65%). PPV were 64% vs. 58% in patients under vs. over 10 years of age. Negative FDG-PET/CT results during follow-up reliably predict the absence of malignancy. Positive FDG-PET/CT scan results in general have a low PPV. The relatively high PPV in patients with histologically proven high grade solid tumors, advanced stages of NHL and with nodular sclerosing, lymphocyte-rich and lymphocyte depleted subtypes of HL warrant a confirmation by biopsy, whereas the watch-and-wait approach can be used in other forms of childhood cancer patients with a positive FDG-PET/CT result in course of follow-up examinations.


1-year pulmonary function and health status in survivors of severe acute respiratory syndrome.
To characterize the long-term pulmonary function and health status in a prospectively identified cohort of patients who survived the severe acute respiratory syndrome (SARS). Prospective follow-up cohort study. University-affiliated hospital. Ninety-four patients who recovered from SARS were assessed at a uniform time point of 1 year after hospital discharge. The study included the measurement of static and dynamic lung volumes, the determination of the diffusing capacity of the lung for carbon monoxide (D(LCO)), and a health status evaluation using the St. George Respiratory Questionnaire (SGRQ). Eleven patients (12%) had mild impairment of FVC, 20 (21%) had mild impairment of FEV1, 5 (5%) had mild impairment of the FEV1/FVC ratio, and 17 (18%) had mild impairment of the D(LCO). There was one patient (1%) who had moderate impairment of FVC, one patient (1%) who had moderate impairment of the FEV1/FVC ratio, and three patients (3%) who had moderate impairment of the D(LCO). No pulmonary function abnormalities were detected in 59 patients (63%). Mean scores were significantly higher (ie, worse) than the population norms in the activity (p < 0.001), impacts (p < 0.001), and total (p < 0.001) domains of the SGRQ. One year after recovery from SARS, persistent pulmonary function impairment was found in about one third of patients. The health status of SARS survivors was also significantly worse compared with the healthy population. The main determinants of morbidity in recovered SARS patients need to be further defined.


OKT3 monoclonal antibody in heart transplantation.
OKT3 monoclonal antibody (OKT3) has already proved to be a valuable edition to the immunosuppression armamentarium available in cardiac transplantation. It is highly effective in treating refractory rejection, where approximately 90% of subjects may be salvaged. It may be even more valuable in prophylaxis, where in combination with an antibody suppression strategy and low-dose, "delayed" cyclosporine it appears to afford near complete protection against rejection. Moreover, OKT3-based prophylaxis seems to impact favorably on the rejection rate after the prophylaxis course has been completed. Adverse reactions are common and generally manageable, although occasional serious clinical events do occur.


Motor cortex rTMS improves dexterity in relapsing-remitting and secondary progressive multiple sclerosis.
The motor cortex (MC) receives an excitatory input from the cerebellum which is reduced in patients with cerebellar lesions. High-frequency repetitive transcranial magnetic stimulation (rTMS) induces cortical facilitation which can counteract the reduced cerebellar drive to the MC. Our study included 24 relapsing-remitting multiple sclerosis (RRMS) and secondary progressive multiple sclerosis (SPMS) patients with dysmetria. The patients were divided into two groups: Group A received two sessions of real MC rTMS and Group B received one session of real rTMS and one session of sham rTMS. Ten healthy volunteers formed group C. Evaluation was carried out using the nine-hole pegboard task and the cerebellar functional system score (FSS) of the expanded disability status scale (EDSS). Group A patients showed a significant improvement in the time required to finish the pegboard task (P = 0.002) and in their cerebellar FSS (P = 0.000) directly after the second session and 1 month later. The RRMS patients showed more improvement than the SPMS patients. Group B patients did not show any improvement in the pegboard task or the cerebellar FSS. These results indicate that MC rTMS can be a promising option in treating both RRMS or SPMS patients with cerebellar impairment and that its effect can be long-lasting.


[2 malignant cell lines derived from strain L6 of rat myogenic cells].
A nonfusing variant M4 of the L6 line of rat myoblasts was isolated. This subline was tumorigenic when inoculated into suckling rats. Comparative studies of cells growth in vitro, serum requirement, cell adhesiveness and capacity to form colonies in soft agar confirmed that M4 cells had undergone neoplastic transformation while L6 cells exhibited normal characteristics. Another subline designated as RMS4 was derivated from M4 cells tumors. Growth characteristics in vitro and karyotype analysis indicated that RMS4 cells presented a higher degree of transformation as compared with M4 cells. Both M4 and RMS4 cell cultures seem to produce a C-type virus.


Identification of Calotropis procera L. as a potential phytoaccumulator of heavy metals from contaminated soils in Urban North Central India.
Lead and cadmium levels were monitored in soil at fifteen urban (riverbank, roadside, industrial and residential) sites in the north central part of India. Calotropis procera, a hardy xerophytic plant was identified and selected for remedial potential as it was seen growing well at all sites. Root and leaf samples were collected simultaneously with soil samples to assess the characteristics of accumulation and tolerance of Pb and Cd in C. procera. Chlorophyll and phenological studies were undertaken to investigate the health of plants. The overall trend of Pb and Cd content in soil and plant samples was in the order Industrial>Roadside>Riverbank>Residential. The highest uptake of both the metals was observed in plants from industrial sites. Sites with more anthropogenic disturbance like vehicular and machinery exhausts exhibited reduced chlorophyll levels, stunted growth as well as a delayed, shortened reproductive phase. The ratios of Pb in leaves to Pb in soil were in the range of 0.60-1.37; while similar ratios of Cd were in the range of 1.25-1.83. Highly significant correlation coefficients were determined between concentrations of Pb and Cd in the samples with R(2) values 0.839 for soil, 0.802 for leaf and 0.819 for root samples. The strong correlation between the degree of contamination and concentrations of Pb and Cd in plant samples identifies C. procera as an effective heavy metal remediator of contaminated lands coupled with environmental stress.


Local area networks and the hospital.
Hospital information systems are characterized by their complexity of individual functions, heterogeneity of functions, and dependence upon integration. A distributed computerized information system is well suited to meeting the needs of hospitals. A local area communications network (LACN) removes a major impediment to the use of distributed systems. An advanced microprocessor-based LACN using fiberoptic communications has been developed by the Applied Physics Laboratory of The Johns Hopkins University and has been implemented at the University of California, San Francisco Hospital.


Clinical staging accuracy and the use of neoadjuvant chemoradiotherapy for cT3N0 rectal cancer: Propensity score matched National Cancer Database analysis.
While neoadjuvant chemoradiation therapy (nCRT) is accepted as standard of care for locally advanced rectal cancer, the approach to treatment of patients with clinically staged T3N0 disease has been increasingly debated. This study examines the accuracy of clinical staging for cT3N0 rectal cancer as recorded in the National Cancer Data Base and evaluates the role of nCRT in treating these patients. Total of 15,843 patients with clinically staged T3N0M0 rectal cancer who either received nCRT or proceeded to surgery-first met inclusion criteria. Propensity score matching was employed to balance the groups. 23% of cT3N0 patients undergoing surgery-first were found to have pathologically positive nodes. Another 16% turned out to have < stage II disease on surgical pathology. Survival curves for matched nCRT and surgery-first groups demonstrated a survival advantage for cT3N0 patients treated with nCRT. Poor clinical staging accuracy can result in both undertreatment and overtreatment of cT3N0 rectal cancer.


Beyond crisis: Enacted sense-making among ethnic minority carers of people with dementia in Australia.
The 'family crisis' narrative is frequently used in dementia studies to explain ethnic minority families' pathways to health and aged care and why there is delayed dementia diagnoses in ethnic minority communities. Such narratives may obscure the family carers' agency in negotiating services and managing personal, social and structural burdens in the lead up to diagnosis. To illuminate agency, this article describes ethnic minority families' pathways to a dementia diagnosis using the concept of sense-making. Three case studies were drawn from 56 video interviews with family carers of older adults with dementia from Chinese, Arab and Indian backgrounds. Interviews were conducted across Australia from February to August 2018, then translated, transcribed and thematically analysed. Findings suggest families did not enter into formal care because of a crisis, instead navigating fragmented systems and conflicting advice to obtain a dementia diagnosis and access to relevant care. This experience was driven by sense-making (a search for plausible explanations) that involved family carers interpreting discrepant cues in changes to the behaviour of the person with dementia over time, managing conflicting (medical) advice about these discrepancies and reinterpreting their relationships with hindsight. The sense-making concept offers a more constructive hermeneutic than the 'family crisis' narrative as it illuminates the agency of carers' in understanding changed behaviours, negotiating services and managing personal, social and structural barriers pre-diagnosis. The concept also demonstrates the need for a multimodal approach to promoting timely diagnosis of dementia in ethnic minority communities through dementia awareness and literacy campaigns as well as initiatives that address structural inequities.


Impact of haptic feedback on surgical training outcomes: A Randomised Controlled Trial of haptic versus non-haptic immersive virtual reality training.
This study aimed to evaluate the educational impact of integrated haptic feedback in an immersive VR bone drilling simulation on the performance of a cohort of junior surgeons. Block randomised, controlled, double-blinded study. St Georges University Hospital, London, United Kingdom. and methods: 31 trainee doctors (postgraduate years 1-3) with limited orthopaedic experience were recruited to participate in this randomised controlled study through e-mail and poster advertising. They were allocated to haptic or non-haptic group through block randomisation prior to entering the study environment. All participants provided verbal and written consent to participate in this study. All participants were blinded to the nature of the study as well as its intervention arms. All participants completed an immersive virtual reality training module with either haptic feedback or no haptic feedback in which they had to drill 3 bicortical holes in a VR tibia bone model in preparation for screw insertion followed by an ex vivo equivalent task on a tibial sawbone model once again drilling 3 holes through both cortices of the tibia. Outcome measures were plunge gap distance, drilling time and objective structures assessment of technical skills (OSAT) as well as qualitative questionnaire outcomes. Haptic feedback in the VR training module showed significantly less plunge gap distance compared to the non-haptic group (7.6 mm ± 4.3 vs 13.6 mm ± 7.4 (p = 0.012)). The haptic group also had longer drill times (17.5 s ± 4.0 vs 13.8 s ± 4.2 (p = 0.027)), higher combined OSAT cores (14 (10,17) vs 8.5 (7.75, 12), p = 0.0006) and greater number of safe drills of <5 mm plunge gap in at least 2 out of 3 attempts (6 (40) vs 0 (0), p = 0.021. This study demonstrates better performance for an orthopaedic surgical task when using a VR-based simulation model incorporating haptic feedback, compared to one without haptic feedback supporting the pursuit and implementation of haptics in surgical training simulation models to enhance their educational value.


1'-Methyl-4'-(4-methyl-phen-yl)dispiro-[1-benzopyran-3(4H),3'-pyrrolidine-2',3''-indoline]-2,2''-dione.
In the title compound, C(27)H(24)N(2)O(3), the pyrroldine ring adopts a twist conformation, while the six-membered pyran-one ring of the coumarin ring system is in a sofa conformation. In the crystal, pairs of N-H⋯O hydrogen bonds link the mol-ecules into inversion R(2) (2)(8) dimers. These dimers are further connected via C-H⋯O hydrogen bonds.


Impact analysis of the discontinuation of reimbursement: the case of oral contraceptives.
We studied the impact of suspending oral contraceptive (OC) reimbursement in the Netherlands for women >21 years starting 1 January 2004. Discontinuation and switching patterns and the time course of the policy intervention's effects were determined. The intervention cohort contained OC users on 1 January 2004; the control cohort users on 1 January 2003. Follow-up duration was 1 year. Discontinuation and switching patterns were assessed using relative risks (RR). Weekly refill fractions were calculated to determine the time course of the policy effects. Our intervention cohort contained 434,917 OC users; the control cohort 489,904 users. When we excluded patients not affected by the policy intervention (i.e., all patients younger than 20 years), discontinuation rates were 15.3% (intervention cohort) and 12.3% (control cohort) (RR=1.24; 95% CI=1.23-1.26) and increased with age. Switching to cheaper OCs was greatest in the intervention cohort, particularly in the 40- to 44-year age group. Differences in cumulative refill fractions showed large variation over time. The OC reimbursement intervention led to an increase in the discontinuation rate of 24%. The effect increased with older age groups. Considering the time course of effects of policy interventions is of critical importance.


Patients who died with steatohepatitis or liver cirrhosis show neuroinflammation and neuronal loss in hippocampus.
Neuroinflammation in the cerebral cortex of patients who died with liver cirrhosis and neuroinflammation, and neuronal death in the cerebellum of patients who died with steatohepatitis or cirrhosis, were reported. Hippocampal neuroinflammation could contribute to cognitive decline in patients with liver disease, but this has yet to be studied. The study aims were to assess if hippocampus from patients who died with steatohepatitis or cirrhosis showed: (i) glial activation, (ii) altered cytokine content, (iii) immune cell infiltration, (iv) neuronal apoptosis and (v) neuronal loss. Post-mortem hippocampus was obtained from 6 controls, 19 patients with steatohepatitis (SH) and 4 patients with liver cirrhosis. SH patients were divided into SH1 (n = 9), SH2 (n = 6) and SH3 (n = 4) groups depending on disease severity. Glial activation, IL-1β and TNFα content, CD4 lymphocyte and monocyte infiltration, neuronal apoptosis and neuronal loss were analyzed by immunohistochemistry. Patients who died in SH1 showed astrocyte activation, whereas those who died in SH2 also showed microglial activation, CD4 lymphocyte and monocyte infiltration, neuronal apoptosis and neuronal loss. These changes remained in patients in SH3, who also showed increased IL-1β and TNFα. Patients who died of liver cirrhosis did not show CD4 lymphocyte infiltration, neuronal apoptosis or increase in TNFα, but still showed glial activation, increased IL-1β and neuronal loss. Patients with steatohepatitis showed glial activation, immune cell infiltration, apoptosis and neuronal loss. Glial activation and neuronal loss remained in cirrhotic patients. This may explain the irreversibility of some cognitive alterations in hepatic encephalopathy. Cognitive reserve may contribute to different grades of cognitive impairment despite similar neuronal loss.


Obstetric and neonatal outcome following minor trauma in pregnancy. Is hospitalization warranted?
To evaluate if hospitalization of pregnant women, involved in minor trauma, for 24h of surveillance, is warranted. The medical files of pregnant women involved in minor trauma, during 2009-2014, at 22-42 gestational weeks, were reviewed. Minor trauma was defined as an injury severity score <3, no immediate complains, normal ultrasound evaluation, reactive non-stress test, and no regular contractions. Patients were divided into those who, according to our departmental protocol, were hospitalized for 24h observation (hospitalized group), and those who refused to be hospitalized, (non-hospitalized group). Pregnancy, delivery and neonatal outcomes were compared between the groups. Included in the study were 946 minor trauma patients that met the inclusion criteria. Gestational age (GA) at the trauma event was lower in the non-hospitalized group (n=331) compared to the hospitalized group (n=615), 29.1 vs. 30.8 weeks, p<0.001, respectively. There were no between-groups differences in the rate of preterm birth, vaginal bleeding, GA at delivery, or cesarean delivery. There were no cases of placental abruption or intrauterine fetal death in both groups. Neonatal outcome did not differ between the groups. Minor trauma during pregnancy, with normal initial assessment, is not associated with adverse pregnancy outcomes. Therefore, routine hospitalization is probably not warranted.


Physical and rheological characterisation of polyethylene glycol-cashew-nut tree gum aqueous two-phase systems.
The characterisation of the polyethylene glycol-cashew-nut tree gum aqueous two-phase system is described. Factors which affect the phase diagram including polymer molecular mass, pH and temperature were analysed. The physico-chemical properties of the system such as density, viscosity, volume ratio and phase separation times were also described. The characteristics of the system studied indicate it to be very attractive as a separation technique.


Parvalbumin-expressing interneurons linearly transform cortical responses to visual stimuli.
The response of cortical neurons to a sensory stimulus is shaped by the network in which they are embedded. Here we establish a role of parvalbumin (PV)-expressing cells, a large class of inhibitory neurons that target the soma and perisomatic compartments of pyramidal cells, in controlling cortical responses. By bidirectionally manipulating PV cell activity in visual cortex we show that these neurons strongly modulate layer 2/3 pyramidal cell spiking responses to visual stimuli while only modestly affecting their tuning properties. PV cells' impact on pyramidal cells is captured by a linear transformation, both additive and multiplicative, with a threshold. These results indicate that PV cells are ideally suited to modulate cortical gain and establish a causal relationship between a select neuron type and specific computations performed by the cortex during sensory processing.


Signatures of granular microstructure in dense shear flows
Granular materials and ordinary fluids react differently to shear stresses. Rather than deforming uniformly, materials such as dry sand or cohesionless powders develop shear bands--narrow zones of large relative particle motion, with essentially rigid adjacent regions. Because shear bands mark areas of flow, material failure and energy dissipation, they are important in many industrial, civil engineering and geophysical processes. They are also relevant to lubricating fluids confined to ultrathin molecular layers. However, detailed three-dimensional information on motion within a shear band, including the degree of particle rotation and interparticle slip, is lacking. Similarly, very little is known about how the microstructure of individual grains affects movement in densely packed material. Here we combine magnetic resonance imaging, X-ray tomography and high-speed-video particle tracking to obtain the local steady-state particle velocity, rotation and packing density for shear flow in a three-dimensional Couette geometry. We find that key characteristics of the granular microstructure determine the shape of the velocity profile.


Modified lyse and wait technique for the treatment of dialysis fistula thrombosis: Percutaneous thrombolytic therapy with a 27-G dental needle.
The lyse and wait technique is a percutaneous endovascular technique in the treatment of thrombosed dialysis fistulas. In this study, our aim was to introduce a more easily applicable modification of the lyse and wait technique and present our results. A total of 84 patients (56 male; 66.6%), with a mean age of 61 years, were included in the study. The treatment was performed in patients presenting with dialysis fistula thrombosis no more than 1 week before the procedure, whose thrombosed segment length was less than 10 cm. A 27-G dental needle was inserted into the thrombosed segment under ultrasonography guidance, and 5 mg of alteplase was injected in a controlled manner. After about 6 to 12 h, the patients were taken to the angiography unit, and balloon angioplasty was performed. The technical and clinical success of the procedure was 98.8%. Technical and clinical failure occurred in only one patient. The median primary patency duration was 13.5 months. None of the patients had major complications. At the end of 12 months, the primary patency rate was 66.6%, and the secondary patency rate was 80%. Our technique has shown to have high technical and clinical success and patency.


[Complications and adverse events in continuous peripheral regional anesthesia Results of investigations on 3,491 catheters].
Over a period of 36 months we prospectively documented infectious, neurological and other complications or adverse events occurring during 3,491 peripheral regional anesthesias via a catheter using computer-based data recording. The investigation included 936 axillar plexus catheters, 473 interscalene plexus catheters, 125 vertical infraclavicular plexus catheters, 74 catheters with psoas compartment blocks, 900 femoral nerve catheters, 964 sciatic nerve catheters and 19 catheters in other localizations. The regional anesthesia catheters were inserted under sterile circumstances (hood, facemask, sterile gloves and coat, surgical disinfection and sterile covering of the placement site) and under peripheral nerve stimulation. 3,070 (87.9%) of the regional anesthesias via catheter, were carried out without any complications. Inflammation (two out of three criteria: redness at insertion site, pain on palpation or swelling) was found in 146 patients (4.2% of all cases). Infections (two out of the criteria: CRP elevation, pus on the insertion site, fever, leucocytosis, necessary antibiotic treatment with exclusion of other possible causes) appeared in 2.4% of all cases (83 patients). In 29 patients (0.8%) we observed severe infections (surgical intervention necessary e.g. abscess incision). Risk factors for inflammation or infections included duration of catheter therapy, cervical localization of the catheter and the experience of the anesthesiologist (p<0.05). Bacterial species most frequently found were Staphylococcus aureus (54%) and Staphylococcus epidermidis (38%). In 0.3% (9 patients) we found short lasting neurological deficits and in 6 patients (0.2%) we recorded a nerve lesion that lasted more than 6 weeks. Other complications occurred in 4.2% of all cases. Special complications such as infections in peripheral catheter regional anesthesia are rare but can pose severe problems. A close postoperative supervision of all regional catheters has to be ensured under careful consideration of the risk factors for infections and the accompanying symptoms.


Interleukin 10 inhibits the rat VSMC proliferation and collagen secretion stimulated by angiotensin II.
To study the effect of interleukin 10 (IL-10) on the angiotensin II (AngII) stimulated rat VSMC proliferation and collagen secretion, and furthermore, explore its mechanism. On cultured VSMC of rat, 3H-thymine (3H-TdR) and 3H-proline incorporations were used to evaluate the DNA and collagen synthesis, respectively. Western blot and immunoprecipitation were applied to assay the expression and activity of focal adhesion kinase (FAK), respectively. IL-10 (10(-8) approximately 10(-10) g/ml) inhibited the increase of 3H-TdR and 3H-proline incorporation as well as FAK activity, which was induced by 10(-7) mol/L AngII (P < 0.05 or P < 0.01). IL-10 also obviously downregulated the synthesis and secretion of collagen by AngII stimulated VSMC. But there was no difference in the protein expression of FAK among all the groups (P > 0.05). IL-10 antagonizes the VSMC proliferation and collagen synthesis by regulating FAK activity stimulated by AngII.


The relationship between ethnography and feminist research.
Ethnography can he viewed both as a method of data collection and as a theoretical or philosophical framework ( 1 ), designed to explore how people interpret their experiences of the world. Feminist research encompasses both quantitative and qualitative methods, and researchers undertake a wide range of strategies and practices within both disciplines.


Hidden Markov Models Capture Behavioral Responses to Suction-Cup Tag Deployment: A Functional State Approach to Behavioral Context.
The biological consequences of behavioral responses to anthropogenic noise depend on context. We explore the links between individual motivation, condition, and external constraints in a concept model and illustrate the use of motivational-behavioral states as a means to quantify the biologically relevant effects of tagging. Behavioral states were estimated from multiple streams of data in a hidden Markov model and used to test the change in foraging effort and the change in energetic success or cost given the effort. The presence of a tag boat elicited a short-term reduction in time spent in foraging states but not for proxies for success or cost within foraging states.


Allosteric controls of nuclear receptor function in the regulation of transcription.
Nuclear receptors control a large number of physiological events through the regulation of gene transcription. Their activation function involves and is controlled by ligands and multiple cofactors (repressors, activators and bridging proteins). Further post-translational modifications resulting from the crosstalk between different signaling pathways provide additional regulations. The numerous players involved allow the sophisticated fine-tuning of transcriptional regulation. The present review describes how allosteric mechanisms are used in the control of the sequential and ordered binding of nuclear receptors and the various protein effectors to target DNA.


Cesarean scar defects and abnormal uterine bleeding: a systematic review and meta-analysis.
To synthesize the published literature to better understand the association between cesarean scar defects (CSDs) and abnormal uterine bleeding (AUB). In particular, we aimed to evaluate the risk and pattern(s) of CSD-associated AUB in addition to exploring the relationship between defect morphology with bleeding symptoms. Systematic review and meta-analysis. Not applicable. Patients with CSD and reports of uterine bleeding as an outcome were identified in 60 studies from database searches. Studies that investigated CSD (as defined by investigators) and reported uterine bleeding, menstrual bleeding, or AUB as an outcome were included. The prevalence and risk of AUB (intermenstrual, postmenstrual, and unscheduled bleeding) in patients with confirmed CSD. Nine studies reported on the prevalence of AUB in patients with a confirmed CSD. Patients with CSD were more likely to experience AUB, compared with those without CSD (relative risk, 3.47; 95% confidence interval [CI], 2.02-5.97; 6 studies, 1,385 patients; I2 = 67%). In a population of patients with at least 1 cesarean delivery, the prevalence of AUB in those with CSD was 25.5% (95% CI, 14.7-40.5; 6 studies, 667 patients, I2 = 93%). However, symptom prevalence was much higher in patients presenting for imaging for a gynecologic indication where the prevalence of AUB in the presence of a CSD was 76.4% (95% CI, 67.8-83.3; 5 studies, 505 patients; I2 = 71%). The mean menstrual duration in symptomatic patients with CSD was 13.4 days (95% CI, 12.6-14.2; 19 studies, 2,095 patients; I2 = 96%), and the mean duration of early-cycle intermenstrual bleeding was 6.8 days (95% CI, 5.7-7.8 days; 9 studies, 759 patients; I2 = 93%). The most common descriptor of CSD-associated AUB was "brown discharge". Patients with larger CSD experienced more bleeding symptoms. There is a strong and consistent association between patients with CSD and AUB. These patients experience a unique bleeding pattern, namely prolonged menstruation and early-cycle intermenstrual bleeding. These data should provide impetus for including CSD as a distinct entity in AUB classification systems. High heterogeneity in our results calls for standardization of nomenclature and outcome reporting for this condition.


Prostaglandin production by minced carrageenin granuloma tissue of rats and its inhibition by dexamethasone and cycloheximide.
Seven-day-old carrageenin granuloma tissue of rats, when incubated in organ culture, produced 6-keto-prostaglandin F1 alpha, prostaglandin E2, prostaglandin F2 alpha, and thromboxane B2 in ratios of 16/7/1/1, respectively. Indomethacin (1 microgram/ml) inhibited this cyclooxygenase-mediated arachidonic acid metabolism. Dexamethasone (1 microgram/ml) also inhibited this metabolic cascade. Its inhibitory effect was not immediate; preincubation of the tissue with dexamethasone potentiated its inhibitory effectiveness. Cycloheximide (0.1 to 10 microgram/ml) also inhibited prostaglandin production. The inhibitory mechanism by cycloheximide of prostaglandin production was discussed in relation to that by dexamethasone.


The trichothiodystrophy syndrome of Pollitt.
Trichothiodystrophy, or sulphur-deficient brittle hair, is a clinical marker for a syndrome that consists primarily of cystine (and hence sulphur)-deficient brittle hair, nail dysplasia, mental and physical retardation and decreased fertility. The radiological aspects of this syndrome have received scant mention in the literature. I describe a 5-year-old boy whose skeleton exhibits axial osteosclerosis and peripheral osteopenia which is similar to two other previously described cases.


Molecular Identification of Phlebotomus caucasicus and Phlebotomus mongolensis (Diptera: Psychodidae) in a Hyperendemic Area of Zoonotic Cutaneous Leishmaniasis in Iran.
Phlebotomus caucasicus Marzinovsky and Phlebotomus mongolensis Sinton are morphologically similar sand fly species. Finding a reliable, fast, and simple method to differentiate these two sand flies is important in understanding their role in the transmission of Leishmania parasite. In our study, 20 specimens of male P. caucasicus, 4 specimens of male P. mongolensis, and 16 specimens of female of both species (Caucasicus group) were examined by polymerase chain reaction (PCR). The result shows identical patterns with a visible fragment of about 500 bp in size. In restriction fragment length polymorphism (RFLP), we observed identical patterns with TasI used as the restriction enzyme. After alignment with sequences of the ITS2 partial gene in GenBank, a perfect match was obtained for the P. mongolensis, but not for P. caucasicus whose sequence was not present in the GenBank. Based on the results of our study, the RFLP-PCR method with nucleotide gene fragment ITS2 was a rapid and reliable method for differentiating these sand fly species.


[Are our medical centres for fitness to drive and firearms licences effective?].
This article aimed to evaluate the effectiveness of medical centres in the assessment of medical fitness for driving and gun licences, as well as describing the differences between them. Using a crossover design and a representative sample of holders of driving and firearms licences in the province of Malaga during 2014 (363 reports for driving licenses and 626 for firearms licenses), an assessment was made of fitness report issued by the centres by comparing it with the records of the Andalusian public health service. The proportion of those that would not meet the legal eligibility requirements was calculated. An analysis was made of the origin of the disagreements as regards the information made available by the centres. The discordance in the assessment of fitness to drive was estimated as 15.4% (95% CI: 12.0 to 19.4), while for firearms licenses it was 2.7 times higher (41.4%). The origin of the discordance is related to the information provided to the centres, rather than the assessment made by them. The limited effectiveness of the centres in the assessment of fitness for driving and, especially, for firearms licenses, could be improved by increased monitoring of their activity, providing health workers with adequate training and access to healthcare records of applicants.


The prevalence of microsporidia in China : A systematic review and meta-analysis.
Microsporidia are a diverse parasite phylum infecting host from all major taxa in all global biomes. This research was conducted to conclude the prevalence of microsporidia in China. All published articles up to February 16, 2018 were considered, including descriptive, cross-sectional, case-control and epidemiology studies. A total of 1052 articles were separated after literature search. After a strict selection according to our criteria, 82 articles were included in qualitative synthesis and ultimately 52 studies were included in quantitative synthesis. Three species of microsporidia were confirmed to exist in China, including Enterocytozoon bieneusi (E. bieneusi), Nosema and Encephalitozoon cuniculi (E. cuniculi). The highest overall estimated prevalence of E. bieneusi in humans was 8.1%, which was observed in acquired immunodeficiency syndrome patients (AIDS). Moreover, the prevalence of E. bieneusi in animals including the cattle, dogs, pigs, deer, sheep and goats were analyszed in this study. The overall estimated prevalence of E. bieneusi acquired by using the random effects model in meta-analysis in cattle, dogs, pigs, sheep and goats and deer was 20.0% (95% confidence intervals: 0.133-0.266, I2 = 98.031%, p < 0.0001), 7.8% (95% CI: 0.050-0.106, I2 = 60.822%, p = 0.0537), 45.1% (95% CI: 0.227-0.674, I2 = 98.183%, p < 0.0001), 28.1% (95% CI: 0.146-0.415, I2 = 98.716%, p < 0.0001) and 19.3% (95% CI: 0.084-0.303, I2 = 96.995%, p < 0.0001) respectively. The overall detection rate of E. bieneusi in water acquired by using the random effects model in meta-analysis was 64.5% (95% CI: 0.433-0.857, I2 = 98.486%, p < 0.0001). Currently, 221 genotypes of E. bieneusi, 1 genotype of E. cuniculi and 6 Nosema were detected in China. The most prevalent genotype of E. bieneusi was genotype D, followed by BEB6 and EbpC.


Low pH fusion of mouse liver nuclei with liposomes bearing covalently bound lysozyme.
Lysozyme covalently bound to liposomes induces the fusion of liposomes with isolated mouse liver nuclei. The fusion behavior is very similar to the case of erythrocyte ghosts (Arvinte, T., Hildenbrand, K., Wahl, P. and Nicolau, C. (1986) Proc. Natl. Acad. Sci. USA 83, 962-966). Kinetic studies showed that membrane lipid mixing was completed within 15 min, as indicated from the resonance energy transfer (RET) measurements. For the resonance energy transfer kinetic measurements the liposomes contained L-alpha-dipalmitoylphosphatidylethanolamine (DPPE), labeled at the free amino group with the energy donor 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) or with the energy acceptor tetramethylrhodamine. The lipid mixing at equilibrium was studied by the fluorescence recovery after photobleaching technique (FRAP). Liposomes (with/without lysozyme) containing Rh-labeled DPPE in their membranes were incubated with nuclei at 37 degrees C, pH 5.2, for 30 min. After washing of nuclei by three centrifugations, 60-70% of the initial amount of labeled DPPE was associated with the nuclei in the case of liposomes bearing lysozyme and only 7-10% in the case of liposomes without lysozyme. For the nuclei incubated with liposomes having lysozyme, about 70% of the total Rh-labeled lipids present in the nuclei diffused in the nuclear membrane(s) (lateral diffusion constant of D = (1.4 +/- 0.5) X 10(-9) cm2/s). By encapsulating fluorescein isothiocyanate-labeled dextran of 150 kDa molecular mass into the liposomes and using a microfluorimetric method, it was shown that after the fusion a part of the liposome contents is found in the nuclei interior. In this lysozyme-induced fusion process between liposomes and nuclei or erythrocyte ghosts, the binding of lysozyme to the glycoconjugates contained in the biomembranes at acidic pH seems to be the determining step which explains the high fusogenic property of the liposomes bearing lysozyme.


Organization of arm movements. Motion is segmented.
A kinematic analysis of human arm trajectories which underlie the production of learned, continuous movements (such as drawing of 'figure 8s' and stars) in free space is presented. The objective of this investigation was to see if a set of rules, which had been identified previously and which are appropriate for generating circular or elliptical motion of the wrist in an arbitrary plane, also hold true for arbitrary, learned trajectories provided one additional assumption is made: that apparently continuous complex movements are composed of unit segments. The results presented in this paper are consistent with this hypothesis. Furthermore, as predicted by the hypothesis, the wrist trajectory deviates little from planar motion in each segment while the plane of motion can change abruptly from one segment to the next.


Quantitative 3D-CT anatomy of hamate osteoarticular autograft for reconstruction of the middle phalanx base.
Hamate osteoarticular autografts are difficult to obtain and it is unclear to what degree the graft matches the joint surface to be replaced and whether a direct ulnar approach might provide a more reliable graft than the standard proximal to distal approach. We modeled hemihamate osteotomies using quantitative three-dimensional CT (3D-CT) to measure the amount of hamate articular surface used and the match with the native volar base of the middle phalanx. In virtual hemihamate osteotomies (standard and direct ulnar) on CTs of 20 patients (11 men and nine women), we measured the percentage of hamate articular surface used for each finger, the match of the articular contour, and the percentage of hamate articular surface removed. The autograft in the standard approach used an average of 26% of the hamate articular surface and had an average 75% match of the articular contour with the volar half of the middle phalanx base. A direct ulnar approach removed an additional small margin of dorsal ulnar hamate with an average maximum width of 2.5 mm and volume of 27 mm(3). An osteoarticular allograft from the hamate to replace the volar half of the middle phalanx base uses less than 1/3 of the hamate articular surface even if the dorsal ulnar margin of the hamate is taken with the graft. These data suggest that it might be feasible to make the deep cut from a direct ulnar approach.


X-linkage, lyonization and a female premium in the verbal IQ results of Orkney schoolchildren, 1947-75.
This paper reports the preliminary findings of an analysis of the Moray House (verbal IQ) results of a base sample of 4383 Orkney children due to be tested in the schools at age 11 +/- 1 1/2 years in 1947-75. Girls enjoy a 3.63% premium relative to phenotypically equivalent boys at test age. Relationships within the sample, which includes some members of the parental generation, are known. The trait is found to be X-linked, with nine phenotypes in the boys, and lyonization is involved.


[Application of numeric integration in waveform morphology of visual evoked potentials in optic nerve diseases. Numeric integration method].
Presentation of new waveform analysis of pattern and flash visual evoked potentials (VEPs) in patients with optic neuropathy. The study involved 48 patients with acute optic neuritis (2nd group), 60 patients with demyelinating (3rd group), 32 patients with compressive (4th group), 24 patients with nonarteritic anterior ischemic optic neuropathy (5th group) and 32 healthy subjects (1st group). Using computer system UTAS E-1000 (pattern and flash stimulation) the VEPs tests were performed. Amplitude, latency of 8 VEP components, 4 gradients, mean waveform and 16 areas of numeric integrals were calculated. Results were analysed with the use of AN-STAT software. Statistical difference recorded under pattern stimulation was obtained in latency of 3rd, 4th, 5th components of VEP in comparing 3th and 4th groups, as well as amplitude of 3rd waveform, second gradient, areas of integrals between cursors number 2 and 4 in comparing 3rd and 5th groups (p < 0.001). Statistical difference recorded under flash stimulation was obtained in latency of 4th component in comparing 2nd and 5th groups, in amplitude of 7th waveform, areas of integrals covering 160 and 224, 224 and 256 ms, between cursors number 2 and 4 in comparing 4th and 5th groups, as well as in a range of areas of integrals covering 160 and 192 ms in comparing 4th and 5th groups (p < 0.001). Results prove clinical potency of the presented new comprehensive method in analysis of VEP in optic nerve diseases.


Antibacterial efficacy of peracetic acid in comparison with sodium hypochlorite or chlorhexidine against Enterococcus faecalis and Parvimonas micra.
The main goal of an endodontic treatment is a complete debridement of the root canal system; however, currently mechanical shaping and chemical cleaning procedures for this purpose have deemed non-satisfactory. The efficacy of peracetic acid (PAA; 0.5, 1.0, 2.0%), as a root canal irrigation solution, against Enterococcus faecalis (DSM 20478) and Parvimonas micra (DSM 20468) when compared with the one of sodium hypochlorite (NaOCI; 1.0, 3.0, 5.0%), chlorhexidine digluconate (CHX; 0.12, 0.2, 2.0%) and 0.9% NaCI (as a control solution) was in vitro investigated with the agar diffusion and direct contact methods. The inhibition zone diameters observed with the agar diffusion test were determined. The viable bacterial counts (CFU/ml) were calculated with the direct method. The agar diffusion test showed that all three root canal irrigation solutions had an efficacy against E. faecalis at all concentrations. The largest inhibition zone diameters against E. faecalis were observed with 5.0% NaOCI. At all three concentrations of PAA, NaOCI, and CHX, the inhibition zone diameter increased with increase in concentration. For P. micra, PAA had a similar inhibition zone diameter despite a concentration increase. In contrast, for NaOCI and CHX, the inhibition zone diameter increased with increasing concentration. 2.0% CHX produced the largest inhibition zone diameter against P. micra. For E. faecalis, only the comparison between 2.0% PAA and 5.0% NaOCI showed statistical significance (p = 0.004). For P. micra the efficacy comparison between the lowest, middle, and highest concentrations of each solution, a statistical significance (p < 0.05) was found for all three solutions. After direct contact with PAA, NaOCI and CHX, no viable bacteria could be determined for either P. micra or E. faecalis. PAA had a similar antibacterial efficacy as the one of NaOCl and CHX when in direct contact with E. faecalis and P. micra. In the agar diffusion test, PAA showed a similar antibacterial efficacy as the one of CHX and a lower one as the one of NaOCl for E. faecalis.


Modifications of plasma and platelet levels of L-DOPA and its direct metabolites during treatment with tolcapone or entacapone in patients with Parkinson's disease.
We compared--retrospectively--the effects of a 3-month therapy with catechol- O-methyltransferase (COMT) inhibitors tolcapone (100 mg, t.i.d.) and entacapone (200 mg, t.i.d.), on L-DOPA metabolism in two groups of parkinsonian patients with motor fluctuations. Plasma and platelets concentrations of L-DOPA and its direct metabolites, dopamine and 3- O-methyldopa (3-OMD), were measured before starting treatment, after two weeks and at the end of treatment. Patients treated with tolcapone showed significant increases in plasma and platelet L-DOPA levels and marked reduction of plasma and platelet 3-OMD levels, both at short- and long-term. Entacapone did not modify L-DOPA levels, while inducing a less marked reduction of plasma and platelet 3-OMD concentrations, with respect to tolcapone, at both time points. Both drugs were similarly effective in increasing plasma and platelet levels of dopamine. These results confirm the different profiles of activity of the two drugs, with tolcapone proving more effective on both the intra- and extra-cellular levels of L-DOPA and 3-OMD.


Neurons containing messenger RNA encoding glutamate decarboxylase in rat hypothalamus demonstrated by in situ hybridization, with special emphasis on cell groups in medial preoptic area, anterior hypothalamic area and dorsomedial hypothalamic nucleus.
Previous deafferentation studies have suggested that most hypothalamic GABAergic innervation originates from neurons within the hypothalamus. We have investigated the distribution of GABAergic cell groups in the rat hypothalamus by means of the in situ hybridization technique, using a cDNA probe for messenger RNA encoding glutamate decarboxylase. Several major GABAergic cell groups were demonstrated, including cells of the tuberomammillary nucleus, arcuate nucleus, suprachiasmatic nucleus, medial preoptic area, anterior hypothalamic area, the dorsomedial hypothalamic nucleus, perifornical area, and lateral hypothalamic area. The most prominent glutamate decarboxylase mRNA-containing cell groups were located in the medial preoptic area, anterior hypothalamic area and dorsomedial hypothalamic nucleus, and were composed of small- to medium-sized neurons. Compared to previously well-characterized GABAergic cell groups in the tuberomammillary nucleus, reticular thalamic nucleus, and non-pyramidal cells of cerebral cortex, the cells of these GABAergic groups demonstrated only weak cDNA labelling, indicating that they contain lower levels of glutamate decarboxylase mRNA. Several types of control experiments supported the specificity of this cDNA labelling, and the GABAergic nature of these cell populations was further supported by detection of glutamate decarboxylase and GABA immunoreactivity. Abundance of GABAergic cells in many hypothalamic nuclei indicates that GABA represents quantitatively the most important transmitter of hypothalamic neurons, and may be involved in neuroendocrine and autonomic regulatory functions.