Effect of aging on the blood-brain barrier.
Aging is commonly associated with progressive deterioration in central nervous system (CNS) function. Nutritional factors or environmental toxins have important effects on CNS degenerative changes. The blood-brain barrier (BBB) is a major modulator of nutrient delivery to the CNS. The tight junctions and the paucity of pinocytosis or fenestrations in brain capillary endothelium act as an effective barrier between the CNS and the circulating toxic agents. Senescence is associated with significant, though often subtle, changes in BBB. Conditions which are commonly associated with aging, such as hypertension and cerebrovascular ischemia, aggravate the age-related alterations in BBB function. The histologic changes in brain vasculature with aging is region selective and species specific. The common age-related histologic changes include loss of capillary endothelial cells, elongation of the remaining endothelial cells, and decreased capillary diameter in rat cortex, but not in the monkey or human cortex, and a decrease in the number of mitochondria in endothelial cells of the brain capillaries in the monkey but not in the rat. The age-related alterations in BBB transport function include a decrease in BBB choline transport with aging and decreased brain glucose influx. The BBB neutral amino acid transport appears to be unaltered in the aged mice. Most of the studies reported so far have failed to show a significant age-related alteration in BBB permeability to water-soluble substances and high molecular weight solutes in the absence of neurological disease. A more profound change in BBB permeability appears to be associated with Alzheimer's disease. Immunohistological studies have demonstrated the presence of serum proteins in the cerebrovascular amyloid in patients with Alzheimer's disease.(ABSTRACT TRUNCATED AT 250 WORDS)


Impact of clinical severity index, infective pathogens, and initial empiric antibiotic use on hospital mortality in patients with ventilator-associated pneumonia.
The prompt initial use of appropriate antibiotics should improve mortality rates in adults with ventilator-associated pneumonia (VAP). However, the incidence of multidrug-resistant (MDR) pathogen infections is on the rise, and the choice of the initial empiric antibiotic may be challenging. We investigated whether appropriate initial antibiotic therapy, infective pathogens, and the clinical severity index influence hospital mortality in patients with VAP and determined independent risk factors for the same. This study evaluated 163 adult patients (aged ≥ 18 years) at Chang Gung Memorial Hospital, Kaohsiung, Taiwan, from January 1, 2007, to January 31, 2008. Eligibility was evaluated based on criteria for VAP. Sequential Organ Failure Assessment (SOFA) scores, Acute Physiological Assessment and Chronic Health Evaluation II (APACHE II) scores, oxygenation index, underlying comorbidities, septic shock status, previous tracheostomy status, and factors related to pneumonia were collected for analysis. Ninety-two patients survived from a total 163 patients with VAP during the course of their confinement in the intensive care unit. Multivariable logistic regression analysis identified that a pre-existing Charlson Comorbidity Index score (P = .011), initial oxygenation index (P = .025), SOFA score (P = .043), VAP caused by Acinetobacter baumanii (P = .030), and infection with MDR pathogens (P = .003) were independent risk factors for hospital mortality in patients with VAP. High Charlson Comorbidity Index score, high initial oxygenation index, high SOFA score, and infection with Acinetobacter baumannii or MDR pathogens significantly affect hospital mortality in patients with VAP.


Identification and molecular characterization of a Bacillus subtilis IS13 strain involved in the biodegradation of 4,5,6-trichloroguaiacol.
4,5,6-Trichloroguaiacol (4,5,6-TCG) is a recalcitrant organochlorine compound produced during pulp bleaching and a potential environmental hazard in paper mill effluents. We report here the identification by biochemical tests and molecular biological analysis, using 16S ribotyping, of a 4,5,6-TCG-degrading bacterium, identified as a strain of Bacillus subtilis that is most closely related according to the phylogenetic analysis to B. subtilis strain Lactipan (alignment score 99%). Biodegradation of 4,5,6-TCG by this organism in a mineral salts medium was shown to occur only when the inoculum was composed of cells in the stationary phase of growth and to be accelerated by an additional carbon source, such as glucose, sucrose, glycerol or molasses. An additional nitrogen source (as ammonium sulfate) did not affect the rate of 4,5,6-TGC removal. No plasmids were detected in the bacterial cells. This is the first strain of B. subtilis which degrades chlorophenols and shows that 4,5,6-TCG is not degraded by cometabolism and that the gene encoding this characteristic is probably located on the chromosome. The lack of requirement for additional nitrogen source, the ability to enhance biodegradation by adding cheap carbon sources such as molasses, and the fact the trait is likely to be stable since it is encoded on the cell chromosome, are all characteristics that make the organism an attractive possibility for treatment of wastes and environments polluted with organochlorine compounds.


Is the subcutaneous route an alternative for administering ertapenem to older patients? PHACINERTA study.
Antibiotic administration by subcutaneous (SC) injection is common practice in French geriatric wards as an alternative to the intravenous (IV) route, but few pharmacokinetic/pharmacodynamic data are available. Ertapenem is useful for the treatment of infections with ESBL-producing enterobacteria. To report and compare ertapenem pharmacokinetic data between IV and SC routes in older persons. Patients >65 years of age receiving ertapenem (1 g once daily) for at least 48 h (IV or SC, steady-state) were prospectively enrolled. Total ertapenem concentrations [residual (C0), IV peak (C0.5) and SC peak (C2.5)] were determined by UV HPLC. Individual-predicted AUC0-24 values were calculated and population pharmacokinetic analyses were performed. Using the final model, a Monte Carlo simulation involving 10 000 patients evaluated the influence of SC or IV administration on the PTA. Tolerance to ertapenem and recovery were also monitored. ClinicalTrials.gov identifier: NCT02505386. Ten (mean ± SD age=87±7 years) and 16 (age=88±5 years) patients were included in the IV and SC groups, respectively. The mean C0 and C2.5 values were not significantly different between the IV and SC groups (C0=12±5.9 versus 12±7.4 mg/L, P=0.97; C2.5=97±42 versus 67±41 mg/L, P=0.99). The mean C0.5 was higher in the IV group compared with the SC group (C0.5=184±90 versus 51±66 mg/L, P=0.001). The mean individual AUCs (1126.92±334.99 mg·h/L for IV versus 1005.3±266.0 mg·h/L for SC, P=0.38) and PTAs were not significantly different between groups. No severe antibiotic-related adverse effects were noted. SC administration of ertapenem is an alternative to IV administration in older patients.


Epidemiology and control of sexually transmitted diseases: strategic evolution.
During the past 10 years, the field of sexually transmitted diseases has evolved from one emphasizing the traditional venereal diseases of gonorrhea and syphilis, to one concerned with the syndromes associated with Chlamydia trachomatis, herpes simplex virus, and human papillomavirus, and then to one preoccupied with the fatal systemic infections caused by human immunodeficiency virus.


PCR sequencing of the spike genes of geographically and chronologically distinct human coronaviruses 229E.
A reverse transcription nested PCR (RT-PCR) sequencing methodology was developed and used to generate sequence data from the spike genes of three geographically and chronologically distinct human coronaviruses 229E. These three coronaviruses were isolated originally from the USA in the 1960s (human coronavirus 229E strain ATCC VR-74), the UK in the 1990s (human coronavirus 229E LRI 281) and Ghana (human coronavirus 229E A162). Upon translation and alignment with the published spike protein sequence of human coronavirus 229E 'LP' (isolated in the UK in the 1970s), it was found that variation within the translated protein sequences was rather limited. In particular, minimal variation was observed between the translated spike protein sequence of human coronaviruses 229E LP and ATCC VR-74 (1/1012 amino acid differences), whilst most variation was observed between the translated spike protein sequence of human coronaviruses 229E LP and A162 (47/1012 amino acid changes). Further, the translated spike protein sequence of human coronavirus 229E A162 showed three clusters of amino acid changes, situated within the 5' half of the translated spike protein sequence.


Two-stage bilateral lung reimplantation in the dog.
A study of two-stage pulmonary autotransplantation was performed on 29 dogs; 12 dogs died after the first stage, which was always a right lung reimplantation, and 17 dogs underwent the second stage after varying intervals of time. In the group of six dogs operated upon with very short intervals between stages (2 to 8 days) there were no survivors. There were seven long-term survivors in the subgroup with an interval longer than 4 weeks, and they constitute the largest published series obtained with this particular procedure. The interval between operative stages was the only significant controlled variable, as only minor technical or operative differences were allowed to occur. Ischemia times of the lungs were usually quite short. The surviving animals were utilized for studies of the regeneration of the afferent pulmonary nerves (Hering-Breure reflex and effects of capsaicin) as permitted by a long follow-up period of up to 3 years.


Integrating transcriptomes and somatic mutations to identify RNA methylation regulators as a prognostic marker in hepatocellular carcinoma.
RNA methylation modifying plays an important role in the occurrence and progression of a range of human cancers including hepatocellular carcinoma (HCC), which is characterized by a mass of genetic and epigenetic alterations. However, the treatment targeting these alterations is limited. We used comprehensive bioinformatics analysis to analyze the correlation between cancer-associated RNA methylation regulators and HCC malignant features in network datasets. We identified two HCC subgroups (cluster 1 and 2), which had clearly distinct clinicopathological, biofunctional and prognostic characteristics, by consensus clustering. The cluster 2 subgroup correlated with malignancy of the primary tumor, higher tumor stage, higher histopathological grade and higher frequency of TP53 mutation, as well as with shorter survival when compared with cluster 1. Gene enrichment indicated that the cluster 2 correlated to the tumor malignancy signaling and biological processes. Based on these findings, an 11-gene risk signature was built, which not only was an independent prognostic marker but also had an excellent power to predict the tumor features. Our study indicated that RNA methylation regulators are vital for HCC malignant progression and provide an important evidence for RNA methylation, methylation regulators are actionable targets for anticancer drug discovery.


Health-Care Providers' Perspectives on Decision-Making Among Seriously Ill Patients of South Asian Origin in the United States.
Persons of South Asian origin (SAs) are the fastest growing minority group in the United States. Culturally competent care for patients and families of SA origin necessitates an understanding of the cultural factors that affect decision-making for palliative and end-of-life care. To better understand health-care providers' perspectives on decision-making among seriously ill persons of SA origin. Data were collected in a US Midwestern city, predominantly in 2 hospital-based health-care systems along with a few interviews of private practitioners. Researchers conducted a thematic analysis of data. Transcripts were coded individually, coding disagreements resolved through discussion and themes arrived upon by consensus. Health-care providers reported that numerous factors interact to influence decision-making among SAs. Patient-based factors include education, time spent in the United States, expectations of physician-directed care, and spiritual beliefs. Family-based factors include a norm of family-based decision-making and a complex distribution of decision-making responsibilities among family members. Provider-based factors include challenges reconciling SA and American approaches to decision-making. Community-based factors include negative attitudes and a lack of education regarding palliative care and the potential role of community support in health-care decision-making. A rich cultural context influences health-care decision-making among seriously ill SAs. An understanding of this context will enhance providers' cultural competence and likely improve services to this growing population.


Peri- and intraventricular cerebral sonography in second- and third-trimester high-risk fetuses: a comparison with neonatal ultrasound and relation to neurological development.
To determine whether periventricular leukomalacia (PVL) and peri- and intraventricular hemorrhages can be detected in fetuses at risk for preterm birth and to establish the clinical significance of this finding. Prenatal cerebral sonography was performed in 26 fetuses at risk for uteroplacental insufficiency on the day of inclusion into the study and weekly until delivery. Neonatal cerebral ultrasound examination was performed within 24 h of birth and biweekly until discharge. The infants underwent standardized neurological examinations. During 30 observations, 21/26 fetuses presented in a cephalic position. Successful visualization in the coronal and sagittal planes was significantly correlated with that of a control population (r = 0.615; 95% CI, 0.390-0.771). In 20/21 fetuses at least one observation was adequate for analysis. Eleven cerebral abnormalities were found in nine fetuses, periventricular echodensities in four, intraventricular hemorrhage in two, and localized thalamic densities in five. In six of these infants ultrasound abnormalities persisted after birth. Neurological follow-up at 24 months demonstrated abnormalities in three infants, two infants were normal and one was lost to follow-up. The three infants with normal ultrasound results after birth had a normal follow-up. Five infants with normal antenatal ultrasound results showed ultrasound abnormalities during the neonatal period; four of them developed PVL Grade 1 and one developed PVL Grade 2. The follow-up of two of these infants was normal, two died and one developed spastic tetraplegia. Abnormal antenatal brain sonography was significantly correlated with gestational age at birth (r = 0.487; P = 0.029). Peri- and intraventricular echodensities can be detected reliably before birth in fetuses at high risk for uteroplacental insufficiency. When abnormalities persist, there is a high risk for an adverse outcome.


Spontaneous cerebellar hemorrhage associated with a novel Notch3 mutation.
A 55-year-old woman with no significant medical history presented with an acute onset severe headache. A non-enhanced CT scan of the head revealed a right cerebellar hemorrhage. Investigation for etiology of the hemorrhage included an MRI showing extensive subcortical ischemic disease and also several previous microbleeds. The MRI appearance and absence of any other etiology for hemorrhage prompted work up for cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). She was found to have a guanosine to thymidine transversion at nucleotide position 1336, codon position 420, resulting in a glycine>cysteine substitution interpreted as "predicted CADASIL-associated mutation". To our knowledge, this mutation has not yet been reported in association with CADASIL.


Transoral robotic salvage oropharyngectomy with submental artery island flap reconstruction.
Transoral robotic surgery (TORS) is an effective approach for select salvage oropharyngeal resections. Reconstruction in these scenarios is often necessary for vascular coverage and optimal functional outcomes but can be challenging. The submental artery island flap (SAIF) is well-suited to this reconstruction. Two patients presented with recurrent oropharyngeal tumors after chemoradiation. Each tumor was resected using TORS, and the SAIF was utilized for reconstruction. Flap inset was completed using TORS in one case. Both patients experienced uneventful recovery free from bleeding events or fistula formation and experienced no partial flap loss or donor site complications. The SAIF is a simple reconstructive option for TORS defects that may be inset using robotic assistance and avoids the need for free tissue transfer.


Cardiovascular emergencies in the pediatric patient.
Children who have heart disease may present to the emergency department (ED) in many stages of life with a range of cardiovascular manifestions, from minimally irritating palpitations to the life-threatening derangements of shock or lethal dysrhythmia. They can present with congenital heart disease, after a temporizing procedure has been performed or after their definitive repair. Children can also present with fever, weakness, dyspnea, syncope, or chest pain; alternatively, children may present to the ED with active dysrhythmia, pulmonary edema, or cardiogenic shock . These symptoms and presentations may result from Kawasaki disease,hypertrophic cardiomyopathy, or arrhythmia; therefore, emergency physicians must also be comfortable with the most common types of heart disease associated with these symptoms and presentations. The purpose of this article is to describe the physiology and presentation of undiagnosed congenital heart disease, to describe the complications that can occur after a staged or definitive repair,and to discuss acquired heart disease in children.


Assessment of the effectiveness of interferential current therapy and TENS in the management of carpal tunnel syndrome: a randomized controlled study.
We assessed the effectiveness of interferential current (IFC) and transcutaneous electrical nerve stimulation (TENS) therapies in the management of carpal tunnel syndrome (CTS) compared with splint therapy, a standard treatment modality for CTS. This was a prospective, single-blinded, single-center, randomized, three-group parallel intervention study of 3 weeks duration. Efficacy was examined in the third week after the end of treatments. Subjects were assigned randomly to one of three groups: group I patients received splint therapy, group II patients received TENS applied on the palmar surface of the hand and the carpal tunnel, and group III patients underwent IFC therapy applied on the palmar surface of the hand and the volar surface of the forearm. TENS and ICF treatments were applied five times weekly for a total of 15 sessions. Group 1 patients were stabilized with volar wrist splints for 3 weeks. The efficacy of the therapies was assessed before initiation of therapy and at 3 weeks after completion of therapy using a visual analog scale (VAS), a symptom severity scale, the functional capacity scale of the BCTQ, and measurement of median nerve motor distal latency (mMDL) and median sensory nerve conduction velocity (mSNCV). Groups were compared pairwise using the Mann-Whitney U test to identify the source of differences between groups. The Wilcoxon test was used to analyze changes in variables over time within a group. In the VAS, BCTQ, MDL, and mSNCV, no significant difference was observed between the groups (p > 0.05). In the VAS, BCTQ, and mSNCV, statistically significant improvements were detected in all groups (p < 0.05). There was no statistically significant difference between TENS and splint therapy with respect to improvement in clinical scores, whereas IFC therapy provided a significantly greater improvement in VAS, mMDL, and mSNCV values than splint therapy (VAS: 4.80 ± 1.18 and 6.37 ± 1.18; p = 0.001, mMDL: 3.89 ± 0.88 and 4.06 ± 0.61; p = 0.001, mSNCV: 41.80 ± 1.76 and 40.75 ± 1.48; p = 0.010). IFC therapy provided a significantly greater improvement in VAS, symptom severity, functional capacity, and mMDL and mSNCV values than TENS therapy (VAS: 4.80 ± 1.18 and 6.68 ± 1.42; p < 0.001, symptom severity: 2.70 ± 1.03 and 3.37 ± 1.21; p = 0.015, functional capacity: 1.90 ± 1.21 and 2.50 ± 0.78; p = 0.039, mMDL: 3.89 ± 0.88 and 4.06 ± 0.88; p = 0.003, and mSNCV: 41.80 ± 1.76 and 41.38 ± 1.78; p = 0.021). IFC may be considered a new and safe therapeutic option for the treatment of CTS.


Assessment of the short-lived non-pure positron-emitting nuclide (120)I for PET imaging.
The non-pure positron-emitting iodine isotope (120)I (T(1/2)=81 min) is a short-lived alternative to (124)I. (120)I has a positron abundance more than twice that of (124)I and a maximum positron energy of 4 MeV. This study was undertaken to evaluate and characterise the qualitative and quantitative PET imaging of (120)I. (120)I was produced via the (120)Te(p,n) reaction on highly enriched (120)Te. The measurements were done with the Siemens scanner HR+ and the 2D PET scanner GE PC4096+. A cylinder containing three cold inserts and a phantom resembling a human brain slice were used to evaluate half-life, positron abundance and background correction. To analyse the image resolution, a -mm tube placed in water was filled with (120)I and (18)F. Comparisons with (18)F, (124)I and (123)I (measured with SPECT) were made using the Hoffman 3D brain phantom. The half-life of 81.1 min was reproduced by the PET measurements. The PET-based positron abundance ranged from 47.9% to 55.0%. The reconstructed image resolution found with the HR+ was 5.4 mm FWHM (12.3 mm FWTM), in contrast to 4.6 mm (8.6 mm) when using (18)F. Erroneous positive and negative numbers of radioactivity found in the cold inserts became nearly zero when the background of gamma-coincidences was corrected for. Images of the Hoffman phantom were inferior to those obtained when (18)F or (124)I was applied but superior to the (123)I-SPECT images. Our data show that (120)I of high radionuclidic purity can be regarded as a suitable nuclide for the PET imaging of radioiodine-labelled pharmaceuticals.


Reconsidering the Edelman equation: impact of plasma sodium concentration, edema and body weight.
Guidelines recommend treatment of dysnatremias to be guided by formulas based on the Edelman equation. This equation describes the relation between plasma sodium concentration and exchangeable cations. However, this formula does not take into account clinical parameters that have recently been associated with local tissue sodium accumulation, which occurs without concurrent water retention. We investigated to what extent such clinical factors affect the Edelman equation and dysnatremia treatment. We performed a post-hoc analysis with original data of the Edelman study. Linear regression was used to examine the effect of age, sex, weight, edema, total body water (TBW) and heart and kidney failure on the Edelman equation. With attenuated correction, we corrected for measurement errors of both variables. Using piecewise regression, we analyzed whether the Edelman association differs for different plasma sodium concentrations. Data was available for 82 patients; 57 males and 25 females with a mean (SD) age of 57 (15) years. The slope of the Edelman equation was significantly affected by weight (p=0.01) and edema (p=0.03). Also, below and above plasma sodium levels of 133 mmol/L the slope of the Edelman equation was significantly different (1.25 x0025vs 0.58x0025, p<0.01). Edelman's equation's coefficients are significantly affected by weight, edema and plasma sodium, possibly reflecting differences in tissue sodium accumulation capacity. The performance of Edelman-based formulas in clinical settings may be improved by taking these clinical characteristics into account.


Pulmonary sequestration and related congenital bronchopulmonary-vascular malformations: nomenclature and classification based on anatomical and embryological considerations.
The pulmonary sequestration spectrum and related congenital lung anomalies present an extremely complex and varied group of bronchopulmonary-vascular malformations. Previous attempts at nomenclature and classification have proved inadequate. In this article we present a classification of the newly named pulmonary malinosculation spectrum, which includes all congenital lung anomalies where there is abnormal connection (that is, malinosculation) of one or more of the four major components of lung tissue--namely, tracheobronchial airway, lung parenchyma, arterial supply, and venous drainage, which in various combinations make up these lesions. We feel that this simple descriptive approach will improve our understanding and management of these complicated lesions and this is supported by the clinical experience we report in the next article.


Allergic reaction in a cohort of haemophilia A patients using plasma-derived factor VIII (FVIII) concentrate is rare and not necessarily triggered by FVIII.
In contrast to haemophilia B, allergic manifestations are rare complications in haemophilia A (HA) patients treated with factor VIII (FVIII) concentrates. Nevertheless, it can be serious and hamper replacement therapy in these cases. The aims of this study were to evaluate the frequency of allergic reaction in a cohort of HA patients treated only with plasma-derived FVIII (pdFVIII) concentrates, and assess the possible immune mechanisms involved. History of allergic reaction was retrospectively assessed. Patients with allergic manifestations were followed, and had plasma samples collected in different timepoints in relation to the allergic episode. These samples were analysed for the presence of inhibitor and anti-FVIII immunoglobulins subclasses. Three of 322 HA patients (0.9%) developed allergic reaction after exposure to pdFVIII products during the last 15 years in our centre. The first patient, with severe HA, without inhibitor, had anti-pdFVIII IgE and IgG4, but no anti-recombinant FVIII (rFVIII) IgE. The second patient, with severe HA, and high-responding inhibitor, presented allergic manifestation with both, pdFVIII concentrate and activated prothrombin complex concentrate. Although anti-pdFVIII and anti-rFVIII IgG4 were detected, no anti-FVIII IgE was present. The third patient, with moderate HA without inhibitor, atopic, had no anti-FVIII immunoglobulin detected, and allergic symptoms disappeared after switching to rFVIII concentrate. This study corroborates the low incidence of allergic reactions in HA patients. In the three cases presented, the anti-FVIII immunoglobulin profile demonstrated that the allergic manifestation was triggered by other proteins contained in pdFVIII products, and not directed to FVIII.


Protein and glycoprotein composition of subsynaptosomal fractions. Implications for Exocytosis and recycling of synaptic vesicles.
Highly enriched fractions of synaptic junctional complexes, synaptic vesicles, and coated vesicles were isolated from rat forebrains and compared, along with synaptosomal plasma membrane and its nonjunctional components, by discontinuous sodium dodecylsulfate-polyacrylamide gel electrophoresis. When stained for proteins with Coomassie blue, the gels all contained the same protein bands, only in different relative amounts. Mixing experiments revealed no additional bands. However, gel patterns of each fraction were not only quantitatively but also qualitatively different when stained for carbohydrate. These observations suggested that some of the protein bands may vary in their degree of glycosylation among the various synaptic fractions. Within the limits of resolution of the methods used here, these results are consistent with the morphological process of synaptic vesicle exocytosis and recycling but suggest the possibility of a reversible modification of certain membrane glycoproteins as they pass through the various membrane compartments.


Bi-national Review of Phaeochromocytoma Care: Is ICU Admission Always Necessary?
Post-operative management after phaeochromocytoma resection includes monitoring of blood pressure and blood sugar, and vigilance for haemorrhage. Guidelines recommend 24 h of continuous blood pressure monitoring, usually necessitating HDU/ICU admission. We hypothesised that most patients undergoing phaeochromocytoma resection do not require post-operative HDU/ICU admission. We aim to describe current Australian and New Zealand perioperative management of phaeochromocytoma and determine whether it is safe to omit HDU/ICU care for most patients. We collected retrospective data on patients undergoing excision of phaeochromocytoma in 12 centres around Australia and New Zealand between 2007 and 2019. Data collected included preoperative medical management, anaesthetic management, vasopressor support, HDU/ICU admission and complications. A total of 223 patients were included in the study, 173 (77%) of whom were admitted to HDU/ICU post-operatively. The group of patients treated in ICU was similar to the group of patients treated on the ward in terms of demographic and tumour characteristics, and there were significant differences in the proportion of patients admitted to HDU/ICU between centres. Of patients admitted to ICU, 71 (41%) received vasopressor support. This was weaned within 24 h in 55 (77%) patients. Patients with larger tumours (> 6 cm) and a transfusion requirement are more likely to require prolonged inotropic support. Among patients admitted to the ward, there were no complications that required escalation of care. Although not widespread practice in Australia and New Zealand, it appears safe for the majority of patients undergoing minimally invasive resection of phaeochromocytoma to be admitted to the ward post-operatively.


Wafer-scale solution-processed 2D material analog resistive memory array for memory-based computing.
Realization of high-density and reliable resistive random access memories based on two-dimensional semiconductors is crucial toward their development in next-generation information storage and neuromorphic computing. Here, wafer-scale integration of solution-processed two-dimensional MoS2 memristor arrays are reported. The MoS2 memristors achieve excellent endurance, long memory retention, low device variations, and high analog on/off ratio with linear conductance update characteristics. The two-dimensional nanosheets appear to enable a unique way to modulate switching characteristics through the inter-flake sulfur vacancies diffusion, which can be controlled by the flake size distribution. Furthermore, the MNIST handwritten digits recognition shows that the MoS2 memristors can operate with a high accuracy of >98.02%, which demonstrates its feasibility for future analog memory applications. Finally, a monolithic three-dimensional memory cube has been demonstrated by stacking the two-dimensional MoS2 layers, paving the way for the implementation of two memristor into high-density neuromorphic computing system.


Antisolvent-assisted Crystallization of Centimeter-sized Lead-free Bismuth Bromide Hybrid Perovskite Single Crystals with X-ray Sensitive Merits.
Hybrid bismuth halides perovskites have emerged as promising candidates for X-ray detection, due to the strong absorptivity of high-energy X-ray photons, high resistivity, large carrier diffusion length and low toxicity. However, the mostly investigated hybrid bismuth iodides single crystals are usually opaque and require a harsh synthesis process. Herein, novel one-dimensional (1D) pentamethylenediamine bismuth bromide (PDA)BiBr5 single crystals were synthesized via an antisolvent-assisted crystallization method at room temperature. Bulk (PDA)BiBr5 single crystals have sizes of 10×1.3×1.5 mm3 and high transparency. They are shown to have low density of defects of 2.0×1010 cm-3 and obvious photoconductivity. Moreover, they exhibit large bulk resistivity of 2.13×1011 Ω cm and good X-ray attenuation coefficient. Consequently, the vertical structured (PDA)BiBr5 single crystal X-ray photoconductor produces a sensitivity of 3.8 μC Gyair -1 cm-2 . This study provides a facile strategy for synthesizing bulk hybrid bismuth bromides single crystals with potential X-ray detection application.


The interplay between the statistical correlations of gamma-ray emission probabilities and efficiency calibration.
The procedure to determine the statistical correlations between gamma-ray intensities is described here, as well as that required to take these correlations into account. The advantages of using branching-ratios and feeding fractions instead of gamma-ray intensities in the gamma-ray standards are discussed.


A composite flap for early treatment of hemifacial microsomia.
A composite flap based on the thoracodorsal pedicle is described for use in the early surgical treatment of children with hemifacial microsomia, having absence or gross hypoplasia of the upper vertical ramus of the mandible and associated soft tissue hypoplasia in the cheek. The flap offers some fascinating possibilities in restoration of function and growth which have yet to be explored.


CAR expression and inducibility of CYP2B genes in liver of rats treated with PB-like inducers.
The expression of the CAR gene and inducibility of CYP2B protein in the liver of male Wistar rats treated with phenobarbital (PB) and triphenyldioxane (TPD) were investigated. To clarify the role of phosphorylation/dephosphorylation in these processes, rats were treated with inhibitors of Ca(2+)/calmodulin-dependent kinase II (W7) or protein phosphatases PP1 and PP2A (OA) before induction. Constitutive expression of the CAR gene in livers of untreated rats was detected by multiplex RT-PCR. Treatment with W7 resulted in a 2.8-fold induction of CAR gene expression, whereas OA led to a 2.4-fold decrease of the mRNA level. The same results were obtained for CYP2B genes expression, which were increased by W7 treatment (two-fold) and decreased by OA (2.3-fold). PB-induction did not lead to significant alteration in the level of CAR gene expression, although CYP2B genes expression was enhanced two-fold over control values. TPD caused a two-fold increase of both CAR and CYP2B mRNA levels. Both inducers reduced the effects of inhibitors on CAR gene expression. Results of EMSA showed that PB, TPD or W7 alone induced formation of complexes of NR1 with nuclear proteins. Appearance of the complexes correlated with an increase in CYP2B expression, and their intensities were modulated by the protein kinase inhibitors. Thus, our results demonstrate that constitutive expressions of CAR as well as CYP2B during induction are regulated by phosphorylation/dephosphorylation processes.


Biotransformation of ginsenosides Re and Rg1 into ginsenosides Rg2 and Rh1 by recombinant β-glucosidase.
Ginsenosides Re and Rg1 were transformed by recombinant β-glucosidase (Bgp1) to ginsenosides Rg2 and Rh1, respectively. The bgp1 gene consists of 2,496 bp encoding 831 amino acids which have homology to the glycosyl hydrolase families 3 protein domain. Using 0.1 mg enzyme ml(-1) in 20 mM sodium phosphate buffer at 37°C and pH 7.0, the glucose moiety attached to the C-20 position of ginsenosides Re and Rg1, was removed: 1 mg ginsenoside Re ml(-1) was transformed into 0.83 mg Rg2 ml(-1) (100% molar conversion) after 2.5 h and 1 mg ginsenoside Rg1 ml(-1) was transformed into 0.6 mg ginsenoside Rh1 ml(-1) (78% molar conversion) in 15 min. Using Bgp1 enzyme, almost all initial ginsenosides Re and Rg1 were converted completely to ginsenosides Rg2 and Rh1. This is the first report of the conversion of ginsenoside Re to ginsenoside Rg2 and ginsenoside Rg1 to ginsenoside Rh1 using the recombinant β-glucosidase.


Systematical evolution on a Zn-Mg alloy potentially developed for biodegradable cardiovascular stents.
To reduce the long-term side effects of permanent metallic stents, a new generation of cardiovascular stents called "biodegradable stents" is being extensively developed. Zinc has been considered as a promising candidate material for biodegradable cardiovascular stents due to its excellent biocompatibility and appropriate biodegradability. However, weak mechanical properties limit its further clinic application. In this study, hot extruded pure Zn and Zn-0.02 Mg alloy were prepared. Compared with pure Zn, Zn-0.02 Mg alloy showed more homogeneous microstructure, much smaller grain size and higher mechanical strength. Zn-0.02 Mg alloy presented uniform corrosion morphologies during the immersion process, and its corrosion rates was higher than that of pure Zn. Hemocompatibility results showed that the Zn-based alloy had extremely low hemolysis rate (0.74 ± 0.15%) and strong inhibitory effect on blood coagulation, platelet adhesion and aggregation. Zn-0.02 Mg alloy also exhibited excellent cytocompatibility. Its extracts could significantly promote the proliferation of endothelial cells. Moreover, the antibacterial activities of the Zn-based alloy were demonstrated by spread plate assay, live/dead viability assay and bacterial morphology observation. These results indicate that the extruded Zn-0.02 Mg alloy has a potential in biodegradable cardiovascular stents.


Pharmacologic considerations for opiate analgesic and nonsteroidal anti-inflammatory drugs.
When administering opioid analgesic drugs or nonsteroidal anti-inflammatory drugs, veterinarians are often not familiar enough with the underlying pharmacology of the drugs, particularly with the potential for drug interactions and adverse effects. This article considers some of the pharmacologic features of these drugs and provides a basis for important interactions, contraindications, and adverse effects.


Serum protein binding kinetics of phenytoin in monotherapy patients.
To determine the binding characteristics of phenytoin to serum proteins in the Japanese population and to compare these with those reported by other investigators. Serum samples examined in the study were obtained from 72 patients (35 males, 37 females) receiving phenytoin monotherapy. The patients' ages ranged from 1 to 73 years (1-15 years, 36 subjects; 16-44 years, 20 subjects; 45-64 years, 13 subjects; > or = 65 years, 3 subjects). The in vivo population binding parameters of phenytoin to serum proteins and theoretical minimal unbound serum phenytoin fraction (fu) were determined using the Scatchard equation. The association constant (K) was 0.020 1/micromol, while the total concentration of binding sites (n(Pt) was 556 micromol/l. The number of binding sites per albumin molecule (n) was 0.85, while binding ability (n.K) was 0.017 l/micromol. The fu was 0.083. The n.K is approximately 1.1 times higher in patients of Pospísil et al. (26) (i.e. 0.0191 l/micromol) than in all our patients. The association constant is approximately 1.1 times higher in our study than in the in vitro study of Monks et al. (23) (i.e. 0-0186 l/micromol), while n is similar between the two studies. The fu in our patients is similar to the unbound serum phenytoin fraction in adult patients receiving phenytoin therapy reported by Richens (2) (i.e. 0.1). Our results suggest that there may be small differences in the binding characteristics of phenytoin to serum proteins between Japanese and non-Japanese subjects. The unbound serum fraction of phenytoin in our patients with epilepsy can be assumed to be relatively constant in the therapeutic concentration range of phenytoin.


Detection of papillomaviral-like DNA sequences in premalignant and malignant perineal lesions of sheep.
Small hyperkeratotic and ulcerated lesions and clinical cancers were isolated from the perineal region of sheep and examined for evidence of papillomavirus infection by various criteria including gross morphology, histology, immunohistochemistry and DNA hybridisation. No specific diagnostic features of papillomaviral infection by immunohistochemistry were found, although some lesions showed gross morphological and histological features similar to papillomaviral effect in other species. DNA hybridisation analysis, using human papillomaviral type 11, 13, 16 and 18 DNA probes under conditions of reduced stringency (Tm-40 degrees C), detected homologous sequences in two thirds of the biopsies examined. These homologous sequences occurred in benign hyperkeratosis as well as invasive squamous cell carcinomas but were much more frequently isolated from carcinomas. This finding suggests that a papillomavirus is associated with the development of squamous cell carcinomas of the perineum of sheep.


Factors affecting clot lysis rates in patients with spontaneous intraventricular hemorrhage.
In the treatment of severe intraventricular hemorrhage (IVH), thrombolytic use and clot size are known to influence clot lysis rates. We evaluated the effect of other variables on IVH clot lysis rates among patients treated with recombinant tissue-type plasminogen activator or placebo. One hundred patients with IVH and intracerebral hemorrhage volume <30 mL requiring emergency external ventricular drainage from 2 multicenter trials were treated with intraventricular administration of recombinant tissue-type plasminogen activator (n=78; 53 males, 25 females) or placebo (n=22; 7 males, 15 females). IVH volume was quantified daily by head CT. A segmented linear regression using an optimized spline knot for each patient was fit. Random effects linear regression was used to estimate the effect of prespecified patient characteristics on clot lysis rates over the first 6 days. Stability IVH volumes were larger in males (N=60; 54 ± 5 mL) than females (N=40; 36 ± 5 mL; P=0.01). Intraventricular thrombolytic treatment was associated with an increase in clot lysis rate of 14.6% of stability IVH volume/day before the spline knot compared with the placebo group (P<0.001). After adjustment for thrombolytic, higher baseline serum plasminogen and lower baseline platelet count were independently associated with an increase in clot lysis of 1.28%/day per 10-g/dL increase (P<0.001) and 0.70% /day per 10×10(3)/uL decrease (P<0.001) before the knot, respectively. Although thrombolysis remains the major determinant of IVH clot lysis rate, higher baseline serum plasminogen and lower platelet count also predict faster clot lysis. Further studies are needed to confirm whether plasminogen availability and thrombus structure impact IVH clot removal. Clinical Trial Registration- URL: http://clinicaltrials.gov. Unique identifier: NCT00650858.


The morpho-genetic and ecological niche analyses reveal the existence of climatically restricted Cycas zeylanica complex in Sri Lanka.
Taxonomy and phylogenesis of Sri Lankan cycad species of the subsection Rumphiae has not been fully resolved and therefore, we conducted an island-wide survey of cycads of the subsection to assess their morphological or genetic variations while exploring the phylogenetic relationship between Sri Lankan Rumphiae and other world cycad species. Further, we assessed the possible distribution of the species in the region through climatic profiling, using maximum entropy modeling approach. We analyzed 21 variable morphological features in collected specimens and used the polymorphism of trnH-psbA locus to understand the phylogeny. The distance tree drawn from the principal component analysis revealed a significant variation in female reproductive structures. The maximum likelihood tree separated Sri Lankan Cycas zeylanica to a well-supported unigeneric clade (bootstrap = 96, posterior probability = 100) with shallow divergence. Ecological niche modeling supported the existence of Cycas zeylanica in South East Asia and in southern Western Ghats in India in addition to the Wet Zone of Sri Lanka. We rename the taxa as Cycas zeylanica complex based on the observed high morphological diversity of female reproductive structures which might have ascended due to multiple introductions of South East Asian cycads by long distance dispersal of seeds through sea currents.


Controlled hatching at the prescribed site using femtosecond laser for zona pellucida drilling at the early blastocyst stage.
To study whether the application of femtosecond laser pulses for zona pellucida (ZP) drilling of blastocysts at the embryonic or abembryonic poles can promote hatching to start immediately through the hole formed and ensure high hatching rates and embryo viability. Mouse blastocyst (E3.5) ZP were microdissected with femtosecond laser pulses (514-nm wavelength, 280-fs pulse duration, 2.5-kHz repetition rate) close to the trophoblast or inner cell mass (ICM). The sizes of the holes formed were in the range of 4.5-8.5 μm. Additional longitudinal incisions (5-7-μm long) on either side of the hole were created to determine whether hatching had started at the correct position. Embryos post-laser-assisted ZP drilling and intact embryos were cultured under standard conditions for 2 days; embryo quality was assessed twice daily. The hatching rates and in vitro and in vivo implantation rates (only for embryos with ZP dissected close to the ICM) were estimated. Femtosecond laser-assisted ZP drilling at the early blastocyst stage facilitated embryo hatching to start at the artificial opening with probability approaching 100%. Despite the artificial opening's small size, no embryo trapping during hatching was observed. Both experimental groups had higher hatching rates than the control groups (93.3-94.7% vs. 83.3-85.7%, respectively). The in vitro implantation rate was comparable with that of the control group (92.3% vs. 95.4%). No statistically significant differences were obtained in the in vivo implantation rates between the experimental and control groups. Blastocyst-stage femtosecond laser microsurgery of ZP is fast and delicate and enables the hatching process to be initiated in a controlled manner through a relatively small opening, with no embryo trapping.


Uterine artery embolization in Tanzania: a procedure with major public health implications.
The burden of uterine fibroids is substantial in sub-Saharan Africa (SSA), with up to 80% of black women harboring them in their lifetime. While uterine artery embolization (UAE) has emerged as an effective alternative to surgery to manage this condition, the procedure is not available to the vast majority of women living in SSA due to limited access to interventional radiology (IR) in the region. One of the few countries in SSA now offering UAE in a public hospital setting is Tanzania. This study aims to assess the safety and effectiveness of UAE in this new environment. From June 2019 to July 2022, a single-center, retrospective cohort study was conducted at Tanzania's first IR service on all patients who underwent UAE for the management of symptomatic fibroids or adenomyosis. Patients were selected for the procedure based on symptom severity, imaging findings, and medical management failure. Procedural technical success and adverse events were recorded for all UAEs. Self-reported symptom severity and volumetric response on imaging were compared between baseline and six-months post-procedure using paired sample t-tests. During the study period, 92.1% (n = 35/38) of patients underwent UAE for the management of symptomatic fibroids and 7.9% (n = 3/38) for adenomyosis. All (n = 38/38) were considered technically successful and one minor adverse event occurred (2.7%). Self-reported symptom-severity scores at six-months post-procedure decreased in all categories: abnormal uterine bleeding from 8.8 to 3.1 (-5.7), pain from 6.7 to 3.2 (-3.5), and bulk symptoms from 2.8 to 1 (-1.8) (p < 0.01). 100% of patients reported satisfaction with outcomes. Among the nine patients with follow-up imaging, there was a mean volumetric decrease of 35.5% (p = 0.109). UAE for fibroids and adenomyosis can be performed with high technical success and low complication rates in a low-resource setting like Tanzania, resulting in significant symptom relief for patients. Building capacity for UAE has major public health implications not only for fibroids and adenomyosis, but can help address the region's leading cause of maternal mortality, postpartum hemorrhage.


[Transmission disequilibrium test of polymorphisms of serotonin transporter gene and schizophrenia based on family trios].
To investigate the relationship between two polymorphisms (Intronic VNTR and 5-HTTLPR) of the serotonin transporter gene and schizophrenia. A set of 314 schizophrenic trio samples collected from Shanghai, Xi'an and Jilin regions of China independently was subjected to analysis of the polymorphisms by transmission/disequilibrium test(TDT). No significantly preferential transmission of any allele was detected from both polymorphisms investigated. The results suggest that the serotonin transporter gene is unlikely to have a major contribution to susceptibility to schizophrenia in Han Chinese population.


The impact of apheresis platelet manipulation on corrected count increment.
Concentrating and washing apheresis platelets (APs) substantially reduce the number of allergic transfusion reactions likely due to removal of plasma. However, these processes may damage platelets (PLTs). This study evaluated whether concentrating or washing APs decrease the corrected count increment (CCI). This retrospective study evaluated individuals who initially received unmanipulated APs and subsequently received concentrated and/or washed APs at a large university hospital between 1998 and 2009. Concentrated units were prepared by reducing the plasma volume of APs by a goal of more than 67%. Washed units were prepared by washing the APs with 1 L of normal saline. The CCI (PLTs [×10(6)] × m(2)/L) for all transfusions was calculated. Hypothesis testing was performed with t tests for continuous variables and chi-square tests for dichotomous variables. We evaluated 121 individuals: 46 patients who received unmanipulated, concentrated, and then washed APs; 59 patients who received unmanipulated and then concentrated APs; and 16 patients who received unmanipulated and then washed APs. Patient demographics were similar among the three groups. The mean CCI for unmanipulated AP transfusions at 0 to 2 hours posttransfusion was significantly higher than concentrated and washed PLT transfusions (p<0.001). When accounting for PLT loss due to manipulation, concentrating APs did not impact the CCI. However, the CCI remained significantly lower for washed products at all time points after transfusion (40.7% mean reduction at 20-24 hr, p<0.001). Washing APs significantly reduces PLT count recovery and survival, as demonstrated by a significantly reduced CCI.


Is the street child phenomenon synonymous with deviant behavior?
The police, court officials, social workers, and the public, in general, perceive street children negatively--their behavior is deemed deviant. This paper examines the concept of deviance as a label placed on the powerless by those in positions of power.


Glucocorticoid repression of the reproductive axis: effects on GnRH and gonadotropin subunit mRNA levels.
Activation of the stress axis by glucocorticoids suppresses reproductive function in many species. Here, we performed studies to determine whether these effects are mediated at the level of the hypothalamus or pituitary or both, and to dissect the underlying molecular mechanisms, using two established rodent models. Rats were treated either chronically or acutely with glucocorticoids, and circulating gonadotropins, GnRH mRNA levels, and gonadotropin subunit mRNAs levels were measured. In model I, chronic treatment for 6 days with corticosterone (CORT) was used in adult intact male rats. CORT caused a significant decrease in serum LH but not FSH secretion compared to vehicle. Whereas pituitary LHbeta and FSHbeta mRNA levels were not affected by CORT treatment, hypothalamic GnRH mRNA was significantly decreased by 35-40%. In model II, acute blockade of the estradiol (E(2))-induced gonadotropin surge by dexamethasone (DEX) was used in 28-day-old female rats. DEX treatment resulted in substantially lower serum LH and FSH concentrations compared to vehicle, although DEX had no effect on GnRH mRNA and LHbeta mRNA levels. By contrast, FSHbeta mRNA levels were about 14-fold lower in DEX-treated females. Taken together, these results indicate that suppression of gonadotropin levels by chronic elevations in glucocorticoids/stress may be accounted for in part by suppression of GnRH mRNA levels, whereas short-term glucocorticoid treatment to block the gonadotropin surge appears to involve other mechanisms including decreased FSHbeta mRNA levels.


Predacious bacteria, Bdellovibrio with potential for biocontrol.
Bacteria of the genus of Bdellovibrio are highly motile Gram-negative predators of other Gram-negative bacteria causing lysis of their prey. Here we report results of studies on the interactions of Bdellovibrio with species of Alcaligenes, Campylobacter, Erwinia, Escherichia, Helicobacter, Pseudomonas, Legionella, and Shigella in agar lower, liquid media and cells attached to a surface. Helicobacter pylori was studied employing both actively growing and viable but nonculturable (VBNC) cells. The majority of the bacterial strains tested were found to be susceptible to Bdellovibrio. A significant observation was that Bdellovibrio attacked both actively growing and VBNC H. pylori, that phenomenon has never been reported. The results indicate that bdellovibrios have potential as biocontrol agents.


Involvement of connexin 43 phosphorylation and gap junctional communication between smooth muscle cells in vasopressin-induced ROCK-dependent vasoconstriction after hemorrhagic shock.
We examined the roles played by gap junctions (GJs) and the GJ channel protein connexin 43 (Cx43) in arginine vasopressin (AVP)-induced vasoconstriction after hemorrhagic shock and their relationship to Rho kinase (ROCK) and protein kinase C (PKC). The results showed that AVP induced an endothelium-independent contraction in rat superior mesenteric arteries (SMAs). Blocking the GJs significantly decreased the contractile response of SMAs and vascular smooth muscle cells (VSMCs) to AVP after shock and hypoxia. The selective Cx43-mimetic peptide inhibited the vascular contractile effect of AVP after shock and hypoxia. AVP restored hypoxia-induced decrease of Cx43 phosphorylation at Ser262 and gap junctional communication in VSMCs. Activation of RhoA with U-46619 increased the contractile effect of AVP. This effect was antagonized by the ROCK inhibitor Y27632 and the Cx43-mimetic peptide. In contrast, neither an agonist nor an inhibitor of PKC had significant effects on AVP-induced contraction after hemorrhagic shock. In addition, silencing of Cx43 with siRNA blocked the AVP-induced increase of ROCK activity in hypoxic VSMCs. In conclusion, AVP-mediated vascular contractile effects are endothelium and myoendothelial gap junction independent. Gap junctions between VSMCs, gap junctional communication, and Cx43 phosphorylation at Ser262 play important roles in the vascular effects of AVP. RhoA/ROCK, but not PKC, is involved in this process.


Collaterals of recurrent laryngeal nerve fibres innervate the thymus: a fluorescent tracer and HRP investigation of efferent vagal neurons in the rat brainstem.
The origin and course of efferent vagal fibers, which innervate the rat thymus, were investigated by a fluorescent retrograde double labeling method, using Fast blue (FB) and Diamidino yellow dihydrochloride (DY) as tracers. In the same animal, one tracer was injected into the cranial portion of the right lobe of the thymus and the other dye was deposited around the cut end of the right recurrent laryngeal nerve. The neuronal population giving origin to the recurrent nerve was mapped by using retrograde labeling with HRP applied to the central stump of the nerve. The HRP retrograde axonal transport showed that most efferent vagal fibers of the recurrent nerve have their perikarya in the nucleus retroambigualis (NRA), nucleus ambiguus (NA), and to a lesser extent in the nucleus retrofacialis (NRF). In fluorescent retrograde double labeling of thymus and recurrent laryngeal nerve both single and double labeled cells were found. The cells labeled by the injections into the thymus were colocalized with the neurons labeled by the tracer deposited in the recurrent laryngeal nerve to the NRA, NA, and NRF. Moreover along the rostrocaudal extent of the NRF and NA double labeled cells were present, showing that some of the thymic efferents are collaterals of the recurrent nerve fibers. Our experiments shown that some thymic vagal fibres originate from neurons of nucleus dorsalis nervi vagi (NDV) as demonstrated both by HRP and FB injected thymuses. The possible role of these efferents in thymic function is briefly discussed.


Preclinical evaluation of radiation and systemic, RGD-targeted, adeno-associated virus phage-TNF gene therapy in a mouse model of spontaneously metastatic melanoma.
The incidence of melanoma in the United States continues to rise, with metastatic lesions notoriously recalcitrant to therapy. There are limited effective treatment options available and a great need for more effective therapies that can be rapidly integrated in the clinic. In this study, we demonstrate that the combination of RGD-targeted adeno-associated virus phage (RGD-AAVP-TNF) with hypofractionated radiation therapy results in synergistic inhibition of primary syngeneic B16 melanoma in a C57 mouse model. Furthermore, this combination appeared to modify the tumor microenvironment, resulting in decreased Tregs in the draining LN and increased tumor-associated macrophages within the primary tumor. Finally, there appeared to be a reduction in metastatic potential and a prolongation of overall survival in the combined treatment group. These results indicate the use of targeted TNF gene therapy vector with radiation treatment could be a valuable treatment option for patients with metastatic melanoma.


Assessing competence in finding and reporting abnormal morphologic features while scanning peripheral blood smears.
To develop a competency assessment tool to determine if technologists are detecting and reporting clinically significant findings on blood smears and to identify if proficiency improves through education using a video presentation. Combination of nonequivalent pretest-post test control group and modified time-series design. A large medical center in Southern Louisiana. All shifts of technologists working in the Core Hematology and Special Hematology Laboratories. This sample is a nonprobability sample of convenience. A thirty-five minute videotape on the standardization of RBC morphology. Develop a method for measuring competency of technologists who perform scans of peripheral blood smears as part of the automated white blood count differential process.


Synthesis and evaluation of a series of benzopyran derivatives as PPAR alpha/gamma agonists.
A series of benzopyran derivatives were synthesized and evaluated for PPAR alpha/gamma agonist activities. Most of the compounds exhibit reasonable PPAR alpha and PPAR gamma agonist activities. In particular, compounds 7b, 8b, 8e and 8h with remarkable PPARg EC(50) values of 0.001microM are excellent full PPAR gamma agonists with the functional potency about 130, 20 times stronger than that of leading compound 5 and rosiglitazone, respectively. Compounds 7a, 7c, 7d and 8a are dual PPAR alpha/gamma agonists, and all of them gave comparable or stronger PPAR alpha/gamma agonist efficacy than that of the corresponding positive control.


Decidual factors and vasoactive intestinal peptide guide monocytes to higher migration, efferocytosis and wound healing in term human pregnancy.
To explore the functional profile of circulating monocytes and decidual macrophages at term human pregnancy and their contribution to tissue repair upon stimulation ex vivo with decidual factors and the vasoactive intestinal peptide (VIP). Peripheral blood monocytes were isolated from pregnant and non-pregnant volunteers and tested in vitro with decidual explants from term placenta and VIP. The effect of VIP on decidual explants and the effect of its conditioned media on monocytes or decidual macrophages isolated by magnetic beads was carried out by RT-qPCR and ELISA for cytokines expression and release. Migration assays were performed in transwell systems. Efferocytosis was assessed in monocytes or decidual macrophages with CFSE-labelled autologous apoptotic neutrophils and quantified by flow cytometry. Monocyte and decidual macrophages wound healing capacity was evaluated using human endometrial stromal cell monolayers. Immunohistochemistry was performed in serial tissue sections of different placentas. VIP is expressed in the villi as well as in trophoblast giant cells distributed within the decidua of term placenta. VIP induced the expression of antiinflmammatory markers and monocyte chemoattractant CCL2 and CCL3 in decidual tissues. Monocytes presented higher migration towards decidual explants than CD4 and CD8 cells. VIP-conditioned monocytes displayed an enhanced efferocytosis and wound healing capacity comparable to that of decidual macrophages. Moreover limited efferocytosis of pregnant women monocytes was restored by VIP-induced decidual factors. Results show the conditioning of monocytes by decidual factors and VIP to sustain processes required for tissue repair and homeostasis maintenance in term placenta.


Role of fetal intertwin difference in middle cerebral artery peak systolic velocity in predicting neonatal twin anemia-polycythemia sequence.
To evaluate the relationship between the fetal intertwin difference in middle cerebral artery peak systolic velocity (MCA-PSV) and intertwin difference in hemoglobin (Hb) concentration at birth in monochorionic diamniotic (MCDA) twin pregnancies in order to assess its potential role in the prediction of twin anemia-polycythemia sequence (TAPS). This was a retrospective cohort study of MCDA twin pregnancies delivered between January 2012 and January 2018. All pregnancies with measurements of MCA-PSV within 7 days prior to delivery and in which neonatal Hb concentration was available were included. The correlation between fetal intertwin difference in MCA-PSV, expressed in multiples of the median (MoM), and neonatal intertwin difference in Hb concentration was investigated. Receiver-operating characteristics (ROC) curve analysis was used to assess the performance of fetal intertwin difference in MCA-PSV for predicting intertwin difference in Hb > 90th centile at birth. A total of 154 out of 256 MC twin pregnancies fulfilled the inclusion criteria. Fetal intertwin difference in MCA-PSV MoM correlated positively with neonatal intertwin difference in Hb concentration (r = 0.79; P < 0.001). The 90th centile for intertwin difference in Hb was 7.25 g/dL. There were 15 (9.7%) cases with a Hb difference ≥ 7.25 g/dL at birth. ROC curve analysis showed a high accuracy of fetal intertwin MCA-PSV MoM difference for the prediction of neonatal intertwin Hb difference ≥ 7.25 g/dL at birth (area under the ROC curve, 0.976 (95% CI, 0.935-0.993); P = 0.012). The optimal cut-off for intertwin MCA-PSV MoM difference was 0.373, with a sensitivity of 93.3% (95% CI, 68.1-99.8%) and a specificity of 95.7% (95% CI, 90.8-98.4%). The positive predictive value was 70% (95% CI, 45.7-88.1%) and the negative predictive value was 99.3% (95% CI, 95.9-100%). Our findings show that fetal intertwin MCA-PSV MoM difference is a good predictor of neonatal intertwin Hb concentration difference > 90th centile and potentially of TAPS. Copyright © 2018 ISUOG. Published by John Wiley & Sons Ltd.


Meibomian Gland Absence Related Dry Eye in Ectodysplasin A Mutant Mice.
Meibomian gland dysfunction is the most frequent cause of evaporative dry eye, yet its underlying pathophysiology is unknown. To gain insight into this pathophysiology, we characterized the time-dependent tear film and ocular surface changes occurring in X-linked anhidrotic-hypohidrotic ectodermal dysplasia mice (Tabby), which lack the meibomian gland. These mice sequentially developed corneal epithelial defects, central corneal stromal edema, neovascularization, and pannus 8 to 16 weeks after birth. Aqueous tear secretion was normal, whereas tear break-up time and ex vivo tear evaporation times were all shortened. Corneal epithelial microvilli were less numerous, conjunctival goblet cell density was unaffected, and MUC5AC and MUC5B gene expression was increased. Markers of squamous metaplasia (cytokeratin 10 and small proline-rich protein 1B) were noticed in the corneal epithelium of Tabby mice as early as the fourth week. Taken together, the Tabby mouse is a relevant meibomian gland dysfunction-related dry eye model that may lead to a better understanding of how meibomian glands are related to ocular surface health. This model may also help with discovering novel drug options for treating evaporative dry eye.


On an integrable multi-dimensionally consistent 2n + 2n-dimensional heavenly-type equation.
Based on the commutativity of scalar vector fields, an algebraic scheme is developed which leads to a privileged multi-dimensionally consistent 2n + 2n-dimensional integrable partial differential equation with the associated eigenfunction constituting an infinitesimal symmetry. The 'universal' character of this novel equation of vanishing Pfaffian type is demonstrated by retrieving and generalizing to higher dimensions a great variety of well-known integrable equations such as the dispersionless Kadomtsev-Petviashvili and Hirota equations and various avatars of the heavenly equation governing self-dual Einstein spaces.


End-of-life considerations in older patients who have lung disease.
The goal of palliative care is to prevent suffering and manage symptoms, maintain quality of life, and provide physical, emotional, and spiritual support for patients and their loved ones. Research suggests that patients with chronic lung disease receive suboptimal palliative care largely because of inadequate communication with their physicians. When patients have made decisions about life-sustaining therapies, physicians often either do not know patients' wishes or misunderstand them. Clinicians should realize that most patients want more information about end-of-life care and that efforts to initiate and improve communication with their patients are important. This article reviews the potential for enhanced palliative care for older patients with chronic lung disease.


Single-Cell RNA Sequencing: Technological Progress and Biomedical Application in Cancer Research.
Single-cell RNA-seq (scRNA-seq) is a revolutionary technology that allows for the genomic investigation of individual cells in a population, allowing for the discovery of unusual cells associated with cancer and metastasis. ScRNA-seq has been used to discover different types of cancers with poor prognosis and medication resistance such as lung cancer, breast cancer, ovarian cancer, and gastric cancer. Besides, scRNA-seq is a promising method that helps us comprehend the biological features and dynamics of cell development, as well as other disorders. This review gives a concise summary of current scRNA-seq technology. We also explain the main technological steps involved in implementing the technology. We highlight the present applications of scRNA-seq in cancer research, including tumor heterogeneity analysis in lung cancer, breast cancer, and ovarian cancer. In addition, this review elucidates potential applications of scRNA-seq in lineage tracing, personalized medicine, illness prediction, and disease diagnosis, which reveals that scRNA-seq facilitates these events by producing genetic variations on the single-cell level.


Californium versus cobalt brachytherapy combined with external-beam radiotherapy for IIB stage cervical cancer: long-term experience of a single institute.
The purpose of this paper was to observe and compare long-term curative effects and complications of FIGO stage IIB cervical cancer patients (n = 232) treated with high-dose-rate (HDR) californium ((252)Cf) neutron or cobalt ((60)Co) photon intracavitary brachytherapy (ICBT) combined with external-beam radiotherapy (EBRT). The EBRT dose to the small pelvis was 50 Gy in both groups. The brachytherapy component of (252)Cf or (60)Co was added in the 3(rd) week of EBRT, 5 fractions were performed once per week resulting in a total ICBT dose of 40 Gy/Gyeq (point A). Overall survival (OS) at 5, 10 and 15 years was 63.6%, 50.4% and 38.8% in the (252)Cf group and 62.2%, 50.5%, 39.9%, in the (60)Co group, respectively (p = 0.74). The percentage of tumour recurrence was statistically significantly lower in the (252)Cf group with 7.4% versus 17.1% in the (60)Co group (p = 0.02). Second primary cancers have developed similarly 9.1% and 8.1% cases for (252)Cf and (60)Co groups, respectively. Our long-term retrospective study comparing (252)Cf and (60)Co isotopes with brachytherapy in combined treatment of FIGO IIB stage cervix carcinoma patients shows, that overall survival in the both groups are similar. However, the recurrence of tumour was significantly lower in the (252)Cf group. The incidence of second primary cancers was similar in both groups.


Cadmium-induced neurotoxic effects on rat basal forebrain cholinergic system through thyroid hormones disruption.
Cadmium (Cd) single and repeated exposure produces cognitive dysfunctions. Basal forebrain cholinergic neurons (BFCN) regulate cognitive functions. BFCN loss or cholinergic neurotransmission dysfunction leads to cognitive disabilities. Thyroid hormones (THs) maintain BFCN viability and functions, and Cd disrupts their levels. However, Cd-induced BFCN damages and THs disruption involvement was not studied. To research this we treated male Wistar rats intraperitoneally with Cd once (1 mg/kg) or repetitively for 28 days (0.1 mg/kg) with/without triiodothyronine (T3, 40 µg/kg/day). Cd increased thyroid-stimulating-hormone (TSH) and decreased T3 and tetraiodothyronine (T4). Cd altered cholinergic transmission and induced a more pronounced neurodegeneration on BFCN, mediated partially by THs reduction. Additionally, Cd antagonized muscarinic 1 receptor (M1R), overexpressed acetylcholinesterase S variant (AChE-S), downregulated AChE-R, M2R, M3R and M4R, and reduced AChE and choline acetyltransferase activities through THs disruption. These results may assist to discover cadmium mechanisms that induce cognitive disabilities, revealing a new possible therapeutic tool.


Case report: renal osteodystrophy in association with spinal stenosis in achondroplasia.
A 54 year old woman with achondroplasia presented with symptoms of spinal stenosis. Subsequent studies revealed end-stage renal disease with severe renal osteodystrophy that was characterised by radiographs and new bone biomarkers. We speculate that renal osteodystrophy accentuated spinal stenosis, which is a common feature of achondroplasia in adults.


Probing the reaction interface in Li-O2 batteries using electrochemical impedance spectroscopy: dual roles of Li2O2.
Li2O2 is revealed to play a positive role by facilitating the kinetics of the oxygen reduction reaction in aprotic Li-O2 batteries, in addition to the well-known negative role by passivating the electrode surface. Moreover, it is inferred that the oxygen reduction reaction takes place at the Li2O2/electrolyte interface, rather than the electrode/Li2O2 interface, in our cell.


Importance of a verification test to accurately assess V̇O2 max in unfit individuals with obesity.
To determine the trustworthiness of graded exercise test to exhaustion (GXT) to assess maximal oxygen uptake ( V ˙ O 2 max ) in metabolic syndrome individuals with obesity and poor cardiorespiratory fitness. V ˙ O 2 max was assessed in 100 metabolic syndrome adults (57 ± 8 years; 34% women), with obesity (BMI 32 ± 5 kg·m-2 ) using GXT followed by supramaximal constant-load verification test (VerT) at 110% of maximal GXT work rate. V ˙ O 2 data from GXT and VerT were compared using paired t test and plotted for Bland-Altman analysis. GXT sensitivity and specificity to detect V ˙ O 2 max were also calculated. Seventy individuals did not achieve V ˙ O 2 plateau during GXT. GXT underestimated V ˙ O 2 max in 40 subjects. In these subjects, the magnitude of V ˙ O 2 max underestimation with GXT was 9% (167 mLO2 ·min-1 ; P < .001). In the whole sample (n = 100), bias error differences between GXT and VerT was 63 mLO2 ·min-1 (3% underestimation). This error was constant regardless of differences in fitness levels among individuals (R = -0.07; homoscedasticity). GXT results were unreliable in 62% of the sample with 16% of false-positive and 46% of false-negative results. Sensitivity and specificity of GTX to assess V ˙ O 2 max were low (ie, 23% and 60%, respectively). Our data indicate that the magnitude (3%-9%) and prevalence (40% of subjects) of V ˙ O 2 max underestimation with the use of a GXT alone is high in a large sample of unfit metabolic syndrome individuals with obesity. Our data advocate for the need of using VerT after GXT to avoid significant cardiorespiratory fitness underestimation in metabolic syndrome individuals with obesity and low fitness level.


Orbital prosthesis following temporal muscle or forehead flap reconstruction: use of optics and illusions.
Temporalis myocutaneous flap reconstruction of orbital defects may cause difficulty with fabrication of esthetic orbital prostheses because of limited space. Two patient treatments are presented to show the wide variation in defect size that may be encountered after surgical reconstruction. Methods for using optics and illusions to enhance the esthetic result are also presented.


Differentiation of cartilaginous anlagen in entire embryonic mouse limbs cultured in a rotating bioreactor.
Mechanisms involved in development of the embryonic limb have remained the same throughout eons of genetic and environmental evolution under Earth gravity (1 g). During the spaceflight era it has been of interest to explore the ancient theory that form of the skeleton develops in response to gravity, and that changes in gravitational forces can change the developmental pattern of the limb. This has been shown in vivo and in vitro, allowing the hypergravity of centrifugation and microgravity of space to be used as tools to increase our knowledge of limb development. In recapitulations of spaceflight experiments, premetatarsals were cultured in suspension in a bioreactor, and found to be shorter and less differentiated than those cultured in standard culture dishes. This study only measured length of the metatarsals, and did not account for possible changes due to the skeletal elements having a more in vivo 3D shape while in suspension vs. flattened tissues compressed by their own weight. A culture system with an outcome closer to in vivo and that supports growth of younger limb buds than traditional systems will allow studies of early Hox gene expression, and contribute to the understanding of very early stages of development. The purpose of the current experiment was to determine if entire limb buds could be cultured in the bioreactor, and to compare the growth and differentiation with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were cultured for six days, either in the bioreactor or in center-well organ culture dishes, fixed, and embedded for histology. E13 specimens grown in culture dishes were flat, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections showed excellent cartilage differentiation in both culture systems, with more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Younger limb buds fused together during culture, so an additional set of E11.5 limb buds was cultured with and without encapsulation in alginate prior to culturing in the bioreactor. Encapsulated limbs grown in the bioreactor did not fuse together, but developed only the more proximal elements while limbs grown in culture dishes formed proximal and distal elements. Alginate encapsulation may have reduced oxygenation to the progress zone of the developing limb bud resulting in lack of development of the more distal elements. These results show that the bioreactor supports growth and differentiation of skeletal elements in entire E13 limb buds, and that a method to culture younger limb buds without fusing together needs to be developed if any morphometric analysis is to be performed.


High-resolution analysis and functional mapping of cleavage sites and substrate proteins of furin in the human proteome.
There is a growing appreciation of the role of proteolytic processes in human health and disease, but tools for analysis of such processes on a proteome-wide scale are limited. Furin is a ubiquitous proprotein convertase that cleaves after basic residues and transforms secretory proproteins into biologically active proteins. Despite this important role, many furin substrates remain unknown in the human proteome. We devised an approach for proteinase target identification that combines an in silico discovery pipeline with highly multiplexed proteinase activity assays. We performed in silico analysis of the human proteome and identified over 1,050 secretory proteins as potential furin substrates. We then used a multiplexed protease assay to validate these tentative targets. The assay was carried out on over 3,260 overlapping peptides designed to represent P7-P1' and P4-P4' positions of furin cleavage sites in the candidate proteins. The obtained results greatly increased our knowledge of the unique cleavage preferences of furin, revealed the importance of both short-range (P4-P1) and long-range (P7-P6) interactions in defining furin cleavage specificity, demonstrated that the R-X-R/K/X-R ↓ motif alone is insufficient for predicting furin proteolysis of the substrate, and identified ≈ 490 potential protein substrates of furin in the human proteome. The assignment of these substrates to cellular pathways suggests an important role of furin in development, including axonal guidance, cardiogenesis, and maintenance of stem cell pluripotency. The novel approach proposed in this study can be readily applied to other proteinases.


Fluorescent high-throughput screening of chemical inducers of neuronal differentiation in skeletal muscle cells.
This protocol describes detailed procedures for the fluorescent high-throughput screening of small molecules that induce neurogenesis in cultures of skeletal muscle cells. The detection of neurogenesis relies on a fluorescent dye, FM 1-43, which is used to study the neuronal property of depolarization-induced synaptic vesicle recycling. Thus, small molecules with neurogenesis-inducing activity in skeletal muscle cells can be rapidly identified by measuring the fluorescence intensity of the treated cells using a fluorescent microplate reader. This protocol uses murine myoblast C2C12 cells for screening, which are readily available and relatively easy to culture. Neurogenesis of PC12 cells induced by nerve growth factor is employed as a positive control for this screening. The screening time for this protocol is 8 d, which also includes the procedure to detect depolarization-induced synaptic vesicle recycling using FM 1-43.


Onset of DNA replication in nuclei of proliferating and resting NIH 3T3 fibroblasts following fusion.
NIH 3T3 mouse fibroblasts arrested in medium containing 0.5% serum were fused with stimulated cells taken at 2-h intervals after replacing the medium with one containing 10% serum, and DNA synthesis was studied in mono-, homo- and heterokaryons using radioautography with double-labelling technique. The presence of a resting nucleus in a common cytoplasm with a stimulated nucleus from the prereplicative period has an inhibitory effect on the entry of the stimulated nucleus into the S period in medium containing either 0.5 or 10% serum, but ongoing DNA synthesis continues. After a 24-h stay in a common cytoplasm with resting nuclei the stimulated nuclei return into the state of rest. When resting cells are stimulated by 10% serum, their inhibitory effect on stimulated nuclei in heterokaryons still persists, at least for 2 h following stimulation. Preincubation of resting cells with cycloheximide for 4 h abolishes their ability to suppress DNA synthesis in stimulated nuclei. The data suggest that resting cells produce an endogenous inhibitor of cell proliferation, whose formation depends upon the synthesis of protein. When stimulated, the cells can proliferate only after decreasing the level of this inhibitor. The results obtained are consistent with the idea of a negative control of cell proliferation.


Reliability of Doppler and stethoscope methods of determining systolic blood pressures: considerations for calculating an ankle-brachial index.
The purposes of this study were to: (1) identify the interrater and intrarater reliability of systolic blood pressures using a stethoscope and Doppler to determine an ankle-brachial index (ABI), and (2) to determine the correlation between the 2 methods. Peripheral arterial disease (PAD) affects approximately 8 to 12 million people in the United States, and nearly half of those with this disease are asymptomatic. Early detection and prompt treatment of PAD will improve health outcomes. It is important that clinicians perform tests that determine the presence of PAD. Two individual raters trained in ABI procedure measured the systolic blood pressures of 20 individuals' upper and lower extremities. Standard ABI measurement protocols were observed. Raters individually recorded the systolic blood pressures of each extremity using a stethoscope and a Doppler, for a total of 640 independent measures. Interrater reliability of Doppler measurements to determine SBP at the ankle was very strong (intraclass correlation coefficient [ICC], 0.93-0.99) compared to moderate to strong reliability using a stethoscope (ICC, 0.64-0.87). Agreement between the 2 devices to determine SBP was moderate to very weak (ICC, 0.13-0.61). Comparisons of the use of Doppler and stethoscope to determine ABI showed weak to very weak intrarater correlation (ICC, 0.17-0.35). Linear regression analysis of the 2 methods to determine ABI showed positive but weak to very weak correlations (r2 = .013, P = .184). A Doppler ultrasound is recommended over a stethoscope for accuracy in systolic pressure readings for ABI measurements.


Motility response to insulin-like growth factor-I (IGF-I) in MCF-7 cells is associated with IRS-2 activation and integrin expression.
In MCF-7L cells, insulin-like growth factor-I (IGF-I) stimulates activation of insulin receptor substrate-1 (IRS-1) and enhances cell proliferation. While others have shown that IGF-I enhances cell motility in MCF-7 cells, we have not been able to demonstrate this. To determine if the source of MCF-7 cells account for these reported differences, we examined the MCF-7 cells available from the American Type Culture Collection (MCF-7/ATCC) and compared them to the MCF-7L cells maintained in our laboratory. Both MCF-7L and MCF-7/ATCC grew in response to 5 nM IGF-I and 1 nM estradiol. However, only MCF-7/ATCC demonstrated IGF-I stimulated motility. Immunoprecipitation of IRS substrates followed by anti-phosphotyrosine blotting demonstrated that both IRS-1 and IRS-2 were activated by IGF-I in these cells. However, MCF-7/ATCC cells had greater phosphorylation of IRS-2 compared to MCF-7L. Immunoblots showed that levels of IRS-1 and IRS-2 were comparable between cell lines. We have previously shown that fibronectin-binding integrins are necessary for IGF-stimulated motility. Similar levels of beta1 integrin were detected in both strains of MCF-7. However, low levels of alpha5 and alpha3 were detected in MCF-7L cells whereas high levels of alpha3 and alpha5 integrin were expressed in MCF-7/ATCC cells. Inhibition of integrin function by a blocking antibody or inhibitory peptide diminished IGF-mediated motility in MCF-7/ATCC. In MCF-7/ATCC cells, IGF-I stimulation was associated with a movement of IRS-2 to the leading edge of filopodia. Thus, patterns of integrin expression among breast cancer cell lines may partially explain the different motility behavior of cells in response to IGF-I. IRS-2 activation and integrin occupancy are both required for IGF-stimulated motility.


Augmentation of cell-mediated cytotoxicity to a rat lymphoma. I. Stimulation of non-T-cell cytotoxicity in vivo by tumour cells.
The injection of a syngeneic Gross-virus-induced lymphoma into W/Fu rats induced peaks of cytotoxicity in the spleen attributable to non-T cells and T cells 3 and 10 days later, respectively. The conditions required for augmenting the cytotoxicity of the non-T cells in various lymphoid compartments (shown elsewhere to closely resemble NK cells) were analysed using the ip and iv routes of inoculation and a variety of tumour cells including those normally susceptible or resistant to lysis by NK cells in vitro. Using an ip inoculation of W/FuG-1 cells (a tumour susceptible to lysis by NK cells), a short-lived, 3-fold increase in cytotoxicity was observed in the spleen at day 3 and a 5-fold increase in the PEC at day 5. Cytotoxicity in other lymphoid organs remained unchanged. Tumours resistant to lysis by NK cells also stimulated cytotoxicity in the spleen or PEC, although the effect depended on the dose and route of inoculation used, and depression of cytotoxicity was observed under some conditions.


Multiplexed Imaging Strategy to Distinguish Indeterminant Biliary Strictures: An Ex Vivo Study.
Indeterminant biliary strictures can be either malignant or benign. Biliary intraepithelial neoplasia (BilIN) is the precursor lesion to cholangiocarcinoma, a deadly bile duct cancer. Current diagnostic methods are limited by inadequate amounts of cells and tissues collected. We aim to demonstrate use of fluorescently-labeled peptides specific for EGFR, claudin-1, and ErbB2 to perform multiplexed imaging of biliary neoplasia. Formalin fixed and paraffin embedded specimens resected from human biliary strictures were sectioned. A gastrointestinal pathologist used standard criteria to score immunohistochemistry from biliary neoplasia and adjacent normal epithelium from the same specimen. Peptides specific for EGFR, claudin-1, and ErbB2 were fluorescently-labeled with FITC, Cy5, and IRDye800, respectively. The fluorophores were chosen to provide spectral separation to distinguish the individual targets. Immuno fluorescence images were collected using confocal microscopy. Target expression was validated using immunohistochemistry. Staining was visualized on the surface of biliary duct epithelial cells and not in the stroma. Greater fluorescence intensity was observed for peptide binding to biliary neoplasia by comparison with normal. The mean ratio for neoplasia-to-normal was 1.4, 1.7, and 1.6, respectively, and the average intensities were significantly greater for neoplasia than normal for each peptide. Peptides and antibody binding co-localized with correlation of ρ=0.64, 0.51 and 0.62, respectively. A panel of fluorescently-labeled peptides can distinguish BilIN and cholangiocarcinoma from normal biliary epithelium, and may be used for multiplexed imaging of indeterminant biliary strictures.


Response to Comments by V. N. Manskikh: "Do External or Internal Factors Lead to Tumor Development? It Is Still Unknown".
The opinion is presented according to which the "bad luck" hypothesis (Tomasetti, C., and Vogelstein, B. (2015) Science, 347, 78-81), which has recently received experimental confirmation, has the right to exist, and its criticisms are largely unfounded.


Altered accumulations of fibronectin are not dependent on fibronectin modifications during the differentiation of F-9 teratocarcinoma stem cells.
F-9 teratocarcinoma stem cells differentiate into parietal endoderm when monolayer cultures are treated with retinoic acid. This change in phenotype is accompanied by increased accumulation and altered organization of fibronectin deposits. Although both stem cells and treated cells synthesize and accumulate fibronectin, only the treated cells deposit a fibrillar array of the protein. We have monitored the accumulation of fibronectin in nontreated and treated F-9 cells with indirect immunofluorescence and have biochemically analyzed the fibronectin synthesized by each cell type with one- and two-dimensional acrylamide gels and peptide maps. Our data suggest that no differences exist between these fibronectins to account for the observed changes in accumulation. Thus, another mechanism may regulate the organization of matrix deposition.


Parasitic Skin Infections for Primary Care Physicians.
The 2 epidermal parasitic skin infections most commonly encountered by primary care physicians in developed countries are scabies and pediculosis. Pediculosis can be further subdivided into pediculosis capitis, corporis, and pubis. This article presents a summary of information and a review of the literature on clinical findings, diagnosis, and treatment of these commonly encountered parasitic skin infestations.


KMnO4 as a probe for lac promoter DNA melting and mechanism in vivo.
The reagent potassium permanganate is used to probe lac transcription complexes by primer extension-probing analysis. A series of strongly hyperreactive bands, corresponding to the known melted region, is observed when open complexes are formed in vitro. A nearly identical pattern occurs in vivo, the signal intensity of which increases when open complexes are trapped with rifampicin. Quantitative comparison of the signal intensity obtained under steady-state conditions with that obtained in the presence of rifampicin indicates that transcription from each of three lac promoter variants is limited in vivo principally by the slow rate of open complex formation. The slow-start lac L8:UV5 promoter is also limited somewhat by slow RNA chain initiation. Slightly different patterns of KMnO4 reactivity at each promoter variant in the absence of RNA polymerase in vitro suggest that DNA sequence dictates the ultimate pattern of melting, with the polymerase acting principally to stabilize the melted state specified by the DNA sequence.


Effect of an amine-fluoride-triclosan mouthrinse on plaque regrowth and biofilm vitality.
The purpose of this double-blind, prospective, latin-square crossover randomised study was to examine the efficacy of a mouthrinse solution containing a combination of triclosan, amine fluoride and sodium fluoride on supragingival plaque regrowth compared to a placebo and a chlorhexidine solution. 12 volunteers refrained after professional oral prophylaxis from all mechanical hygiene measures for the following 96 h and rinsed instead cross-over in a randomised order with either chlorhexidine digluconate (0.2%, positive control), a verum solution (0.5% amine fluoride, 0.028% sodium fluoride, 0.03% triclosan) or a placebo solution. The plaque index was assessed after 24 and 96 h (PI 1, PI 2) and the plaque area (PA) photographed and calculated after 96 h of undisturbed plaque regrowth. Moreover plaque samples were taken after 24 and 96 h and examined with the vital fluorescence technique to assess the vitality of the biofilm microbiota (VF 1, VF 2). The amine-fluoride-triclosan solution reduced the clinical parameters (PI and PA) as well as the vitality of the plaque flora (VF) significantly when compared to the placebo solution. The verum reached a 36.5% (p<0.05) and a 39.8% reduction (p<0.0001) in PI 1 and PI 2, respectively, concomittant with a reduction of 23.8% and 32.2% (p<0.001) in VF 1 and VF 2 and of 46.9% (p<0.0001) in PA at day 4. This was less than the reductions found with the positive control, i.e., the 0.2% chlorhexidine solution (54.2% and 71.1% reduction in PI 1 and PI 2, 40.0% and 53.4% in VF 1 and VF 2 and 71.5% in PA). However, significant differences between both active solutions were only established for PI 2 and PA. During 4-day plaque regrowth the amine-fluoride-triclosan product displayed a significant antibacterial and plaque-reducing action in comparison to the control.


A clinically validated method to separate and quantify underivatized acylcarnitines and carnitine metabolic intermediates using mixed-mode chromatography with tandem mass spectrometry.
Acylcarnitines are intermediate metabolites of the mitochondria that serve as biomarkers for inherited disorders of fatty acid oxidation and amino acid metabolism. The prevailing clinical method used to quantify acylcarnitines involves flow-injection tandem mass spectrometry, an approach with a number of limitations; foremost the inability to separate and therefore distinguish key isobaric acylcarnitine species. To address these issues, we report a clinically validated liquid chromatography tandem mass spectrometry method to quantify acylcarnitines, free carnitine, and carnitine metabolic intermediates in human plasma. Importantly, this method resolves clinically relevant isobaric and isomeric acylcarnitine species in a single 22 min analysis without the use of ion pairing or derivatization reagents. This unique combination of features is not achievable by existing acylcarnitine methods and is made possible by the use of a novel mixed-mode chromatographic separation. Further clinical validation studies demonstrate excellent limits of quantification, linearity, accuracy, and inter-assay precision for analyses of 38 different calibrated analytes. An additional 28 analytes are semi-quantitatively analyzed using surrogate calibrators. The study of residual patient specimens confirms the clinical utility of this method and suggests expanded applicability to the diagnosis of peroxisomal disorders. In summary, we report a clinically validated acylcarnitine method that utilizes a novel mixed-mode chromatographic separation to provide a number of advantages in terms of specificity, accuracy, sample preparation time, and clinical utility.


Home parenteral nutrition in children: 8 years of experience with 112 patients.
It is essential that children on prolonged parenteral nutrition for anatomical or functional loss of small bowel should enjoy a quality of life which is as normal as possible. Their return home is a major factor in this. Over the past 8 years, 112 children were able to remain at home on cyclic parenteral nutrition. Forty-nine of them are no longer on home parenteral nutrition (HPN), 45 are still on HPN, and 18 have died. Growth and quality of life were good in most cases. Most of the complications were from infection, 1 septicaemia per 594 days on HPN. In the light of these results, HPN seems to be the best option for children requiring prolonged parenteral nutrition, although it can only be considered within the framework of a specialized centre, which ensures patient follow-up, and provides the logistical support required for this high-technology treatment.


Management of recurrent epistaxis in an anticoagulated patient by temporarily closing the nares with sutures.
We describe an unusual case of recurrent, refractory anterior epistaxis in an 86-year-old man with two mechanical heart valves who was on permanent warfarin therapy. His numerous episodes of epistaxis were incited by chronic nose-picking and strong nose-blowing, practices that he continued to engage in despite repeated medical advice to stop. Stopping his anticoagulation therapy was not considered as a management option because of an unacceptably high risk that this would lead to a thromboembolic event. Eventually, we temporarily sutured his nares closed, and his nosebleeds ceased. The suturing was performed in the ward with local anesthesia. This procedure was simple to perform,fairly well tolerated, easily reversible, and highly effective.


Impact of body mass index on relapse in children with acute lymphoblastic leukemia treated according to Nordic treatment protocols.
High body mass index (BMI) is associated with poorer survival in childhood acute lymphoblastic leukemia (ALL), but the actual impact on the risk of relapse still needs to be clarified. We evaluated the impact of BMI at diagnosis on the risk of relapse in children with ALL treated according to Nordic Society of Paediatric Haematology and Oncology (NOPHO) protocols. In a multicenter study, we collected data on BMI at diagnosis and outcome of 2558 children aged 2.0-17.9 years diagnosed between 1992 and 2016. Patients were divided into four groups according to International Obesity Task Force (IOTF) childhood BMI cut-offs: underweight, <17; healthy weight, 17-25; overweight, 25-30; and obese, ≥30 kg/m2 . In Cox multivariate regression analyses, an increased risk of relapse was observed in children aged 10-17.9 years with unhealthy BMI at diagnosis (underweight hazard ratio HR: 2.90 [95% confidence interval: 1.24-6.78], P = .01; overweight, HR: 1.95 [1.11-3.43], P = .02, and obese HR: 4.32 [95% 2.08-8.97], P < .001), compared to children with healthy weight. BMI had no impact on relapse in children under 10 years of age. High BMI, and especially obesity at diagnosis, is an independent adverse prognostic factor for relapse in older children with ALL.