Serum uric acid levels and incidence of impaired fasting glucose and type 2 diabetes mellitus: a meta-analysis of cohort studies.
A meta-analysis of cohort studies was conducted to assess the association between serum uric acid (SUA) levels and incidence of impaired fasting glucose (IFG) and type 2 diabetes mellitus (T2DM). A comprehensive search was conducted to identify eligible studies. The fixed or random effect pooled measure was selected based on between-study heterogeneity. Dose-response relationship was assessed by restricted cubic spline model and multivariate random-effect meta-regression. Twelve studies with fifteen results were included involving 6340 cases and 62,834 participants. The pooled multivariate-adjusted relative risk (RR) (95%CI) of IFG and T2DM for the highest vs. lowest level of SUA was 1.54 (1.41-1.68), I(2)=42.2%. The association was consistent and significant across subgroup analysis. A nonlinear relationship was found of SUA levels with incidence of IFG and T2DM (P<0.01), and the multivariate-adjusted RRs (95%CI) of IFG and T2DM were 1.02 (0.95-1.10), 1.04 (0.94-1.15), 1.10 (0.99-1.22), 1.25 (1.16-1.35), 1.43 (1.31-1.55), 1.50 (1.38-1.63) and 1.49 (1.34-1.67) for 2.5, 3.5, 4.5, 5.5, 6.5, 7.5 and 8.5mg/dl of SUA. The RR (95%CI) of T2DM for the highest vs. lowest level of SUA was 1.67 (1.51-1.86), and a nonlinear relationship was also found between SUA levels and incidence of T2DM. SUA levels are positively associated with incidence of IFG and T2DM, and the association might be nonlinear.


[The reverse effect of sexualsteroids on the serum-lysozyme (author's transl)].
75 women received different preparates of contraceptives (Non-Ovlon, Ovosiston, Gravistat, Deposistion--VEB Jenapharm Jena, GDR) for two years. In the last month of the application of the sexual steroids and one month later the lysozyme titer in the sera was tested to determine a reverse or stop-effect. An elevation of the muramidase could be shown in all cases. This findings are interpreted in relation to a non manifest influence of sexualsteroids on the lysozyme.


Human S100A12: a novel key player in inflammation?
S100A12 is a member of the S100 family of EF-hand calcium-binding proteins. Human S100A12 is predominantly expressed and secreted by neutrophil granulocytes and, therefore, has been assigned to the S100 protein subfamily of calgranulins or myeloid-related proteins. Intracellular S100A12 exists as an anti-parallel homodimer and upon calcium-dependent activation interacts with target proteins to regulate cellular functions. Extracellular S100A12 exists majorily as homodimer and hexamer, respectively, and shows cytokine-like characteristics. It is part of the innate immune response and linked to certain autoimmune reactions. Human S100A12 is markedly overexpressed in inflammatory compartments, and elevated serum levels of S100A12 are found in patients suffering from various inflammatory, neurodegenerative, metabolic, and neoplastic disorders. In this regard, interaction of calcium-activated S100A12 with the multiligand receptor for advanced glycation endproducts (RAGE) and its soluble form (sRAGE) plays a central pathogenetic role. Recent clinical evidence suggests a high potential of S100A12 as a sensitive and specific diagnostic marker of localized inflammatory processes.


Long Non-Coding RNA SPRY4-IT1 Can Predict Poor Prognosis in Digestive System Malignancies: a Meta-Analysis.
Recent studies have reported that long non-coding RNA SPRY4 intron transcript 1 (SPRY4-IT1) is abnormally expressed in malignant digestive tumors and associated with prognosis. But its clinical relevance is unclear. Here, we performed a meta-analysis aims to evaluate the prognostic value of SPRY4-IT1 in digestive system malignancies. We systematic search the PubMed, Web of Science, ScienceDirect and Wiley Online Library database to eligible studies. The pooled hazard ratios (HRs) with a 95% confidence interval (95% CI) were calculated to explored the association of lncRNA SPRY4-IT1 with prognosis. Five studies were eligible for analysis, a total of 518 patients were included. Meta-analysis indicated that overexpression of SPRY4-IT1 was associated with poor over survival (OS) (HR = 1.24, 95%CI:0.49-1.98; random-effects model). The clinicopathological parameters analysis further showed that increased expression level of SPRY4-IT1 was positively correlated with lymph node metastasis (HR = 1.45, 95% CI =0.88-2.02; fixed-effects model), TNM stage (HR = 1.24,95% CI = 0.78-1.70; fixed-effects model), and invasion depth (HR = 1.25,95% CI = 0.63-1.88; fixed-effects model). lncRNA SPRY4-IT1 may serve as a potential prognostic marker in malignant digestive tumors.


The growth equation of cities.
The science of cities seeks to understand and explain regularities observed in the world's major urban systems. Modelling the population evolution of cities is at the core of this science and of all urban studies. Quantitatively, the most fundamental problem is to understand the hierarchical organization of city population and the statistical occurrence of megacities. This was first thought to be described by a universal principle known as Zipf's law1,2; however, the validity of this model has been challenged by recent empirical studies3,4. A theoretical model must also be able to explain the relatively frequent rises and falls of cities and civilizations5, but despite many attempts6-10 these fundamental questions have not yet been satisfactorily answered. Here we introduce a stochastic equation for modelling population growth in cities, constructed from an empirical analysis of recent datasets (for Canada, France, the UK and the USA). This model reveals how rare, but large, interurban migratory shocks dominate city growth. This equation predicts a complex shape for the distribution of city populations and shows that, owing to finite-time effects, Zipf's law does not hold in general, implying a more complex organization of cities. It also predicts the existence of multiple temporal variations in the city hierarchy, in agreement with observations5. Our result underlines the importance of rare events in the evolution of complex systems11 and, at a more practical level, in urban planning.


Comparison of "click-to-sense" assay with frozen section analysis using simulated surgical margins in breast cancer patients.
We have shown that "Click-to-sense" (CTS) assay based on the visualization of cancer cells by fluorescence probe targeted for acrolein is useful for differentiating between the malignant and benign lesions of the breast. In the present study, we aimed to apply CTS assay to the examination of the simulated surgical margins, being compared with frozen section (FS) analysis. The simulated surgical margin samples (n = 300) were obtained from 1 to 2 cm distant sites from the tumor margin in the mastectomy specimens of breast cancer patients, and divided into the training (n = 150) and validation (n = 150) set. The samples were subjected to CTS assay, subsequently to FS analysis and finally to permanent section (PS) analysis. Diagnostic accuracy of the CTS assay and FS analysis was evaluated in the examination of the simulated surgical margin status finally determined by the PS analysis. In the training set, sensitivity, specificity, and accuracy was 89.3%, 98.4%, and 96.7% for the CTS assay and 89.3%, 98.4%, and 96.7% for the FS analysis. In the validation set, sensitivity, specificity, and accuracy was 93.3%, 98.3%, and 97.3% for the CTS assay, and 93.3%, 99.2%, and 98.0% for the FS analysis. The CTS assay is as accurate as the FS analysis in the examination of the simulated surgical margins in breast cancer patients, and it seems to have a potential to replace the FS analysis for the intra-operative examination of surgical margins in breast-conserving surgery since it is less labor-intensive and more time-saving than the FS analysis.


PRMT2 accelerates tumorigenesis of hepatocellular carcinoma by activating Bcl2 via histone H3R8 methylation.
Protein arginine methyltransferases (PRMTs) have been implicated in the development of various cancers. PRMT2, a member of the type I PRMT family, is overexpressed in multiple tumors. However, the expression and role of PRMT2 in hepatocellular carcinoma (HCC) have not been studied. Here, we discovered that PRMT2 expression is elevated in HCC tissues compared to the corresponding non-tumor tissues, and PRMT2 overexpression is an independent predictor of poor prognosis in HCC patients. Depletion of PRMT2 in HCC cell lines inhibited their cell growth and induced apoptosis. Mechanistic investigations showed that PRMT2 is responsible for H3R8 asymmetric methylation (H3R8me2a). H3R8me2a enrichment at the Bcl2 promoter increases its accessibility to STAT3, promoting Bcl2 gene expression. In addition, our results confirmed that the catalytically inactive mutant of PRMT2 or the type I PRMT inhibitor MS023 impaired the pro-tumorigenic functions of PRMT2 in HCC cells. Overall, our findings showed that PRMT2 functions as an oncogenic gene in HCC, revealing its potential as a novel therapeutic target in HCC.


Use of dual-energy X-ray absorptiometry (DEXA) to follow mineral content changes in small ceramic implants in rats.
Dual energy X-ray absorptiometry (DEXA) imaging was used to quantify bone ingrowth into hydroxyapatite (HA) ceramic implants in rats. HA cylinders implanted in the proximal tibiae were followed for 13 wk. The increase in alkaline phosphatase (ALP) activity within the implants preceded the increase in mineral content as measured non-invasively by the DEXA technique. This was consistent with the timing of ALP activity in respect of mineralization as they occur during fracture healing. The results show that DEXA imaging is useful in measuring bone ingrowth into small ceramic HA implants in vivo, despite the high mineral content background of the implant scaffold.


[Mechanisms of EGF regulation of COX-2 through the STAT5 signaling pathway in human lung adenocarcinoma A549 cells].
It has been proved that cyclooxygenase-2 (COX-2) is a key factor in lung cancer oncogenesis. COX-2 can be induced by a number of cytokines and growth factors and can be regulated by the JAK/STAT signaling pathway. Inhibiting the expression of COX-2 can prevent the development of lung cancer. The aim fo this study is to investigate whether the epidermal growth factor (EGF) can stimulate the signal transducers and activators of transcription 5 (STAT5) as well as to discover the effects of the STAT5 signaling pathway on the COX-2 in human lung adenocarcinoma A549 cells. The phenomenon of STAT5 activation stimulated by the EGF was assayed through immunofluorescence and Western blot. The adenovirus containing the wild-type (WT)-STAT5 (AdWT-STAT5) plasmid, dominant-negative (DN)-STAT5 (Ad-CMV5Stat5aΔ740) plasmid, and STAT5 siRNA were transfected into A549 cells. The latter two groups were stimulated using EGF. Reverse transcriptase polymerase chain reaction was used to detect the mRNA expression of COX-2. STAT5 was not activated in A549 cells in vitro. EGF stimulation significantly increased the level of the p-STAT5 protein and induces the shuttling of p-STAT5 from the cytoplasm into the nucleus. STAT5 activation was crucial for the COX-2 expression induced by the EGF. STAT5 was required for COX-2 expression, but can mediated the effects of the COX-2 expression through pathways that were independent of transcriptional activation. COX-2 expression is dependent on STAT5 phosphorylation. A second pathway does not require STAT5 phosphorylation.


Computerized tomography in chronic subdural hematomas (effusions) of infancy.
During the past 2 years six infants were seen at University of Wisconsin Hospitals (UWH) with subdural hematomas (effusions). This communication describes the abnormalities found on computerized tomographic (CT) evaluation of the six infants.


Monocyte-mediated suppression of IL-2-induced NK-cell activation. Regulation by 5-HT1A-type serotonin receptors.
In the present study, the effects of serotonin on human natural killer (NK)-cell responsiveness to interleukin 2 (IL-2) was investigated. Concomitant treatment of human lymphocytes, enriched for NK effector cells by Percoll density-gradient centrifugation, with serotonin and IL-2 yielded a synergistic activation of NK-cell cytotoxicity (NKCC) in the presence but not in the absence of monocytes. The monocyte-dependent, regulatory effects of serotonin and/or IL-2 were prostaglandin-independent and could be reconstituted when monocytes, recovered by countercurrent centrifugal elutriation (CCE), were added to purified NK effector cells. The effects of serotonin on baseline and IL-2-activated NK cells were mimicked by the 5-HT1A receptor-specific agonists 8-OH-DPAT and (+)-ALK. Our data suggest that serotonin regulates NK-cell responsiveness to IL-2 via 5-HT1A receptors.


Breaking the symmetry: assembly of cylindrical nanostructures with a C3-symmetrical ligand.
The reaction of a tripodal ligand containing terminal 2,3-dihydroxypyridine groups with (arene)ruthenium(II) complexes resulted in the formation of cylindrical nanostructures.


Characterization of monoclonal antibodies directed against trail or trail receptors.
A subset of tumour necrosis factor receptor family members is involved in death transducing signals and is, therefore, referred as the "death receptors." Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in many tumour cells but only rarely in normal cells. Five distinct receptors have been described for TRAIL: TRAIL R1 (DR4), TRAIL R2 (DR5, TRICK), TRAIL R3 (TRID, DcR1), TRAIL R4 (TRUNDD, DcR2), and osteoprotegerin. In the Eighth International Workshop on Human Leukocyte Differentiation Antigens, 10 monoclonal antibodies (mAbs) reported to be specific for TRAIL or for TRAIL receptors were submitted. In the present study, the mAb specificity was determined by ELISA. Using these mAbs, investigation on the expression of TRAIL and TRAIL receptors was performed. Some of them were able to modulate TRAIL induced programmed cell death.


The predictive value of resting electrocardiograms for 12-year incidence of coronary heart disease in the Honolulu Heart Program.
The predictive value of electrocardiographic (ECG) abnormalities at baseline for subsequent risk of fatal and total coronary heart disease (CHD) was analyzed in a 12-year follow-up of 7682 men in the Honolulu Heart Program aged 45-68 who were free of CHD at baseline. Univariate analysis showed that men with major or minor ECG abnormalities, ST depression, high R wave, T-wave inversion, left ventricular hypertrophy or strain and premature ventricular contractions had considerably higher (2-10 fold) CHD incidence rates than those with normal ECG. When blood pressure, cigarette smoking, body mass index, alcohol use, physical activity, serum glucose, serum cholesterol, serum uric acid, age and years lived in Japan were taken into consideration through multivariate analysis, the ECG abnormalities retained significant associations with fatal and total CHD. This study demonstrated that resting ECG abnormalities were independent predictors of both total and fatal CHD.


Widdrol-induced lipolysis is mediated by PKC and MEK/ERK in 3T3-L1 adipocytes.
Obesity is a serious medical condition causing various diseases such as heart disease, type-2 diabetes, and cancer. Fat cells (adipocytes) play an important role in the generation of energy through hydrolysis of lipids they accumulate. Therefore, induction of lipolysis (breakdown of lipids into fatty acids and glycerol), is one of the ways to treat obesity. In the present study, we investigated the lipolytic effect of widdrol in 3T3-L1 adipocytes and its mechanism. Widdrol considerably increased the amount of glycerol released from 3T3-L1 adipocytes into the medium in a time- and dose-dependent manner. To determine the mechanism of this effect, we investigated the alterations in glycerol release and protein expression in 3T3-L1 adipocytes treated with widdrol alone or widdrol and inhibitors of proteins involved in the cAMP-dependent pathway or cAMP-independent PKC-MAPK pathway, which are known to induce lipolysis in adipocytes. The adenylyl cyclase inhibitor SQ-22536, PLA2 inhibitor dexamethasone, PI3K inhibitor wortmannin, and PKA inhibitor H-89, which were used to investigate the involvement of the cAMP-dependent pathway, did not affect the lipolytic effect of widdrol. Widdrol-induced phosphorylation of PKC, MEK, and ERK, which are related to the PKC-MAPK pathway, and their phosphorylation was inhibited by their inhibitors (H-7, U0126, and PD-98059, respectively). Moreover, the increase in glycerol release induced by widdrol was almost completely blocked by PKC, MEK, and ERK inhibitors. These results suggest that widdrol induces lipolysis through activation of the PKC-MEK-ERK pathway.


Benign paroxysmal positional vertigo in children.
To describe the clinical features of benign paroxysmal positional vertigo (BPPV) in children. A retrospective study. Six children diagnosed with BPPV between March 2014 and March 2015 were retrospectively evaluated. BPPV was diagnosed using the Dix-Hallpike and supine roll tests and treated with either the modified Epley particle repositioning procedure or Lempert or Gufoni manoeuvre. Follow-up was performed at 1-week intervals until vertigo and nystagmus disappeared during positional testing. A total of six children were followed up for a period of 10-22 months. Clinical features such as history, nystagmus and symptoms of vertigo, dizziness and nausea. Six children were diagnosed with BPPV using positional testing and treated with the modified Epley or Lempert/Gufoni particle repositioning procedures. Four children were diagnosed with posterior canal BPPV, while the remaining two were diagnosed with horizontal canal BPPV. One girl reported a history of head trauma, one girl had a family history of vertigo, and one boy reported hearing loss in the same ear as that affected by BPPV. Overall, 83.33% of children (5/6) were completely relieved of vertigo following one treatment session. The remaining child was asymptomatic after two sessions. No child reported relapse of vertigo during the follow-up period. BPPV can be diagnosed accurately by taking a detailed medical history and by use of positional testing. BPPV in children can be successfully identified and treated.


Determination of Dapoxetine Hydrochloride in Human Plasma by HPLC-MS/MS and Its Application in a Bioequivalence Study.
Dapoxetine is used for the treatment of premature ejaculation. The present study developed an HPLC-MS/MS method to determine the levels of dapoxetine in human plasma processed using simple protein precipitation. Dapoxetine-d7 was selected as the internal standard. The established method was performed using a mass spectrometer equipped with an electrospray ionization source in multiple positive ion reactions to monitor the mode using the precursor-to-product ion transitions of m/z 306.2-157.2 and m/z 313.2-164.2 for dapoxetine-d7 and dapoxetine, respectively. The method was evaluated based on its selectivity, linearity, limit of quantification, precision, accuracy, matrix effects, dilution integrity, stability, and extraction recovery. As a result of the model used in the present study, the validated linear ranges of dapoxetine were determined to be 2.00~1000 ng/mL in plasma, and the selectivity, precision, accuracy, dilution integrity, stability, and extraction recovery met the accepted standard. No matrix interference was observed. The method was successfully validated and applied to pharmacokinetic studies in healthy Chinese volunteers during the fasting and postprandial periods, respectively.


Association of APOE polymorphisms with disease severity in MS is limited to women.
The authors studied the association of an exon 4 (E4*epsilon2/3/4) and three promoter polymorphisms of APOE with disease course and severity stratified by gender in 221 patients with multiple sclerosis from two overlapping population-based prevalence cohorts. Women carriers of the E4*epsilon2 allele took longer to attain an Expanded Disability Status Scale score of 6 (p = 0.015) and had more favorable ranked severity scores than noncarriers (p = 0.009). There was no association in men. Alleles epsilon3 or epsilon4 and promoter polymorphisms were not associated with disease course or severity.


Gold nanoparticle as an alternative tool for a urine pregnancy test.
The urine pregnancy test is an easily available diagnostic test in the present day and is routinely performed. The test is based on an immunochromatography technique. Here, we used an advanced nanomedicine technique for modification of the urine pregnancy test. The preparation of gold nanoparticle solution in this study followed the standard method. We performed an experiment on both pregnancy-positive and -negative urine samples. First, a mixture with an equal amount (500 microliters) of gold nanoparticle solution and urine sample was prepared. Then, it was further tested for pregnancy by the urine pregnancy test strip. The pregnancy-positive mixture became pink, while the pregnancy-negative mixture became gray. The urine pregnancy test strip for a positive mixture had two lines, while the negative mixture had one line. This application can help the diagnosis of pregnancy and can be an alternative method for a urine pregnancy test. To our knowledge, this is the first report on this application.


Simultaneous quantitative analysis of letrozole, its carbinol metabolite, and carbinol glucuronide in human plasma by LC-MS/MS.
Letrozole is an efficient endocrine treatment of postmenopausal breast cancer, however, not all patients benefit from this treatment, and moreover, severe side-effects like arthralgia frequently lead to discontinuation. To better understand inter-individual variability in drug response and side-effects, plasma analysis of steady-state concentrations of letrozole and its major metabolites is crucial. We developed a rapid, sensitive, and specific method for the simultaneous quantification of letrozole and its metabolites 4,4'-(hydroxymethylene)dibenzonitrile (carbinol) and bis(4-cyanophenyl)methyl hexopyranosiduronic acid (carbinol-gluc) by UHPLC-ESI-MS/MS using in-house synthesized, stable isotope-labeled internal standards. Following solid-phase extraction in BondElut C18 96-well plates, the analytes were separated on a ZORBAX Eclipse XDB-C18 column (1.8 μm, 4.6 × 50 mm) with a gradient of acetonitrile in 0.1% acetic acid in water and detected on a triple quadrupole mass spectrometer with electrospray ionization in the multiple reaction monitoring mode. Lower limits of quantification were 20, 0.2, and 2 nM for letrozole, carbinol, and carbinol-gluc, respectively. The assay has been validated according to FDA guidance and applied to the analysis of 20 plasma samples of postmenopausal breast cancer patients treated with 2.5 mg of letrozole per day. Mean plasma levels (±SD) were 366 ± 173, 0.38 ± 0.09, and 34 ± 12 nM for letrozole, carbinol, and carbinol-gluc, respectively. Our rapid and sensitive mass spectrometry based method enables future pharmacokinetic investigations of letrozole outcome.


Anti-inflammatory properties and pharmacological induction of Hsp70 after brain injury.
The 70-kDa heat shock protein (Hsp70) is thought to protect the brain from a variety of insults. Although the mechanism has been largely limited to its chaperone functions, recent work indicates that Hsp70 also modulates inflammatory pathways. Brain injury and ischemia are associated with an immune response that is largely innate. Hsp70 appears to suppress this response and lead to improved neurological outcome. However, most of this work has relied on the use of genetic mutant models or Hsp70 overexpression using gene transfer or heat stress, thus limiting its translational utility. A few compounds have been studied by various disciplines which, through their ability to inhibit Hsp90, can cause induction of Hsp70. The investigation of Hsp70-inducing pharmacological compounds has obvious clinical implications in terms of potential therapies to mitigate neuroinflammation and lead to neuroprotection from stroke or traumatic brain injury. This review will focus on the inflammation modulating properties of Hsp70, and the current literature surrounding the pharmacological induction in acute neurological injury models with comments on potential applications at the clinical level.


Separation of chiral nanotubes with an opposite handedness by chiral oligopeptide adsorption: A molecular dynamics study.
The separation of enantiomeric chiral nanotubes that can form non-covalent complexes with an unlike stability upon adsorption of chiral molecules is a process of potential interest in different fields and applications. Using fully atomistic molecular dynamics simulations, we report in this paper a theoretical study of the adsorption and denaturation of an oligopeptide formed by 16 chiral amino acids having a helical structure in the native state on both the inner and the outer surface of the chiral (10, 20) and (20, 10) single-walled carbon nanotubes having an opposite handedness, and of the armchair (16, 16) nanotube with a similar diameter for comparison. In the final adsorbed state, the oligopeptide loses in all cases its native helical conformation, assuming elongated geometries that maximize its contact with the surface through all the 16 amino acids. We find that the complexes formed by the two chiral nanotubes and the chosen oligopeptide have a strongly unlike stability both when adsorption takes place on the outer convex surface of the nanotube, and when it occurs on the inner concave surface. Thus, our molecular simulations indicate that separation of chiral, enantiomeric carbon nanotubes for instance by chromatographic methods can indeed be carried out using oligopeptides of a sufficient length.


Clinical Characteristics and Outcomes of Children and Adolescents With Cancer Infected With SARS-CoV-2 at a Tertiary Care Medical Center in Jordan.
Our knowledge about severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is still evolving; its effects on children with cancer need to be studied more. The aim of this study is to present our experience with SARS-CoV-2 infection in this population and to highlight specific complications and outcomes. This is a retrospective and prospective observational study, which involved 21 cancer patients below the age of 18 years in north Jordan. Data regarding their age, sex, cancer type and progression, phase of treatment, and others were collected and reviewed. Patients were classified into confirmed, probable, and suspect according to the Centers for Disease Control and Prevention's (CDC) 2021 classification. A total of 21 patients with malignancy were included. Ten patients were males (48%). Mean age of 8.8 years (3 mo to 18 y). Two patients (9.5%) had died; one (4.7%) death was coronavirus disease 2019 (COVID-19)-related and the other one (4.7%) was due to cancer progression. Two patients (9.5%) had multisystem inflammatory syndrome in children. Both disease progression and new malignancies were documented in 11 (52%) of our patients. Diagnosis of COVID-19 should not distract physicians from investigating new malignancy or relapse as they may come together or may be related to COVID-19 infection. More studies are needed to identify the contribution of SARS-CoV-2 in the pathogenesis of cancer.


Overflow in science and its implications for trust.
To explore increasing concerns about scientific misconduct and data irreproducibility in some areas of science, we interviewed a number of senior biomedical researchers. These interviews revealed a perceived decline in trust in the scientific enterprise, in large part because the quantity of new data exceeds the field's ability to process it appropriately. This phenomenon-which is termed 'overflow' in social science-has important implications for the integrity of modern biomedical science.


Treatment results of 84 patients with nasopharyngeal carcinoma in childhood.
To evaluate the clinical characteristics, treatment regimens, survival, and late complications in patients with nasopharyngeal carcinoma. Eighty-four patients diagnosed with nasopharyngeal carcinoma between 1972 and 2002 were retrospectively evaluated. Tumor status, lymph node involvement, metastasis, and stage were assessed according to the TNM classification. All patients were treated with radiotherapy and chemotherapy. Chemotherapeutic regimens involved cyclophosphamide from 1972 to 1982, vincristine + cyclophosphamide + epirubicine and actinomycin-D protocol from 1982 to 1999 and cisplatin + methotrexate + 5-fluorouracil with leucovorine rescue after 1999. Mean and median values were used for the demographic characteristics. Kaplan-Meier survival curves were used for survival analysis. The patient groups were compared in terms of survival duration using a log-rank test. There were 51 males and 33 females with a median age of 13 years (7-17) at diagnosis. At the time of diagnosis, 78.5% of the patients were at advanced stage (stages III and IV). Overall and event-free survival rates were 56.3% and 50.1%, respectively. Survival was inversely affected by advanced nodal status (P = 0.01). The survival rates (80%) were found higher with cisplatin + methotrexate + 5-fluorouracil + leucovorine rescue regimen. The most frequent late complications after therapy were hypothyroidism (9.5%), dental caries (7.1%), and trismus (7.1%). The cisplatin-based regimen was the best chemotherapeutic regimen. Less toxic regimens with better survival rates should be investigated in patients with nasopharyngeal carcinoma.


Risk factors for decreased total body and regional bone mineral density in hemodialysis patients with severe secondary hyperparathyroidism.
Hyperparathyroidism contributes significantly to decreased bone mineral density (BMD) in end-stage renal disease patients, but this negative influence is not homogeneous throughout the skeleton. We studied the BMD by dual-energy X-ray absorptiometry on total body and on different regions of the skeleton in 42 patients with severe hyperparathyroidism on hemodialysis. We also evaluated the relationship between different risk factors and BMD found on the regions examined in these patients. The legs and other sites where cortical bone predominate were mostly affected, whereas trabecular bone was relatively preserved. This is probably the result of the different effects of hyperparathyroidism on cortical and trabecular bone, but we cannot rule out the interference of ectopic calcifications and sclerotic lesions of vertebral end-plates falsely increasing lumbar spine BMD. The main determinants of low total-body BMD were, in order of importance, immobility, high intact parathyroid hormone levels, low body mass index, and low albumin. Eleven patients presented with pathologic fractures, mainly in the legs, and BMD was lower in this group than in patients without fractures. In conclusion, our study makes clear that hyperparathyroidism is a great threat to bone density in hemodialysis patients, mainly in the legs, the site mostly affected by fragility fractures in our patients. Physicians must worry not only with high parathyroid hormone levels, but also with the nutritional state of these patients.


Staphylococcus aureus utilizes environmental RNA as a building material in specific polysaccharide-dependent biofilms.
Biofilms are surface-bound microbial communities that are typically embedded in a matrix of self-produced extracellular polymeric substances and can cause chronic infections. Extracellular DNA is known to play a crucial role in biofilm development in diverse bacteria; however, the existence and function of RNA are poorly understood. Here, we show that RNA contributes to the structural integrity of biofilms formed by the human pathogen Staphylococcus aureus. RNase A dispersed both fresh and mature biofilms, indicating the importance of RNA at various stages. RNA-sequencing analysis demonstrated that the primary source of RNA in the biofilm matrix was the Brain Heart Infusion medium (>99.32%). RNA purified from the medium promoted biofilm formation. Microscopic and molecular interaction analyses demonstrated that polysaccharides were critical for capturing and stabilizing external RNA in biofilms, which contributes to biofilm organization. These findings provide a basis for exploring the role of externally derived substances in bacterial biofilm organization.


Evaluation of left ventricular diastolic function by tissue Doppler imaging in patients with newly diagnosed and untreated primary generalized epilepsy.
The effects of epilepsy and seizures on cardiac functions have been documented, and this association has drawn attention in recent years. The aim of this study was to investigate left ventricle (LV) diastolic function by tissue Doppler imaging (TDI) in patients with newly diagnosed and untreated primary generalized epilepsy (PGE). Thirty newly diagnosed and untreated PGE patients (14 females, mean age 27.60±9.64 years) and 30 healthy age- and gender-matched control subjects (14 females, mean age 29.47±6.89 years) were included in the study. The LV functions of the study cohort were evaluated using conventional echocardiography and TDI. There were no significant differences found between the two groups regarding the left atrium diameter, left atrium volume index, interventricular septum and posterior wall thickness, LV end diastolic diameter, and LV end diastolic volume (p>0.05 for each parameter). PGE patients exhibited a higher LV end systolic diameter and LV end systolic volume compared to the controls (p<0.001 for each parameter). Thus, the fractional shortening and ejection fraction were lower in the PGE patients (p<0.001 for each parameter). The E to average e' ratio, the most important noninvasive indicator of LV filling pressure, was significantly higher in patients with PGE (8.31±2.78 vs. 6.95±1.26, p=0.018). The present study reports the systolic and diastolic dysfunction of LV in newly diagnosed PGE patients compared to control subjects. Taken together, the screening of epileptic patients using conventional echocardiography and TDI may be useful to evaluate the effects of epilepsy on cardiac functions.


Are you with me? The relevance of peers and other social resources for participation in the German voluntary accompanied-driving scheme.
In Germany, "Accompanied Driving From Age 17" (AD17) is a voluntary alternative method of formal driver training that demonstrably reduces the crash risk for young novice drivers. This study aims to clarify whether social resources such as peers and novice driver-specific social capital play a role in participation in the AD17 program in Germany. In a nationwide random sample, 17-year-old accompanied novice drivers (n = 2,478) and 18-year-old novices (n = 925) who had conventionally acquired a driver's license were surveyed about their novice driver-specific social resources, using specially developed measurement tools such as a resource generator. The results revealed that the proportion of a young driver's friends currently taking or having taken part in AD17 is a major predictor for participation in AD17. To a lesser extent, family-based social capital positively correlates to voluntary participation in AD17. Future optimization efforts should therefore take into account providing support for family-based social resources, including the possibility of compensation. Additionally, considering the composition of the young driver's peer group is strongly recommended.


Circ_0020397 regulates the viability of vascular smooth muscle cells by up-regulating GREM1 expression via miR-502-5p in intracranial aneurysm.
Circ_0020397 has been found to be down-regulated in intracranial aneurysm (IA), and deregulation of circ_0020397 involved in the regulation of vascular smooth muscle cells (VSMCs) proliferation. However, the mechanism by which circ_0020397 implicates in VSMC dysfunction in IA remains vague. The expression of circ_0020397, miR-502-5p and Gremlin 1 (GREM1) was detected using quantitative real-time polymerase chain reaction. Cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Protein levels of proliferating cell nuclear antigen (PCNA) and GREM1 were measured using western blot. The interaction between miR-502-5p and circ_0020397 or GREM1 was confirmed by dual-luciferase reporter, RNA pull-down and RNA immunoprecipitation assay. Circ_0020397 or GREM1 expression was decreased in VSMCs isolated from IA patients, and overexpression of circ_0020397 or GREM1 promoted VSMC viability and elevated PCNA expression level, while inhibition of them showed opposite effects. MiR-502-5p was confirmed to directly bind to circ_0020397 or GREM1, and miR-502-5p reversed the effects of circ_0020397 on VSMC viability and PCNA level. Besides, miR-502-5p overexpression suppressed VSMC viability and reduced PCNA level, while these effects were attenuated by GREM1 up-regulation. Importantly, circ_0020397 could regulate GREM1 expression via miR-502-5p in VSMCs. Circ_0020397 played an important role in phenotypic modulation in IA by promoting VSMC viability via miR-502-5p/GREM1 axis, suggesting a novel insight into IA pathogenesis and new targets for IA molecular therapy.


Operative Treatment of Symptomatic Meso-Type Os Acromiale.
Os acromiale occur frequently in the population and can be a cause of significant shoulder dysfunction and rotator cuff impingement. When symptomatic os acromiale fail conservative management, surgical intervention may become necessary. The purpose of this article is to describe our preferred surgical technique for osteosynthesis of os acromiale using cannulated screws and tension-band wiring.


An improved method for the identification of Gc1 subtypes (group-specific component) by isoelectric focusing.
The occurrence of common subtypes of the Gc1 gene in a German population (n = 261) was investigated by isoelectric focusing on thin layers of polyacrylamide in the pH range 4-6. Six common phenotypes designated Gc 1S, 1F, 1F-1S, 2-1S, 2-1F, and 2 are considered as gene products of three common alleles with the following frequencies: Gc1S=O.603, Gc1F=0.125, and Gc2=0.272. The sum of the allele frequencies of Gc1S and Gc1F corresponds with that of the 'old' Gc allele. Family investigation are in agreement with an autosomal codominant way of inheritance. the method employed provides an identification of the subtypes within 4 h without using anti-Gc-serum for immunoprecipitation.


A Rasch model analysis of technology usage in Minnesota hospitals.
To identify whether the level of difficulty varied among various healthcare technologies. In addition, to examine the whether the degree of healthcare technology adoption was related to patient safety. The data on healthcare technology usage came from a survey of hospitals in Minnesota. There were responses from individuals within 104 hospitals for an effective response rate of 72.7%. The data on patient safety was taken from the Hospital Compare database. Rasch model analysis and regression analysis were used to analyze the data. Rasch model analysis revealed that the difficulty of implementation of healthcare information technologies varied by the particular technology. That is, some technologies were more difficult than other technologies. Further, it was found that the degree of healthcare information technology implementation within a hospital was significantly related to patient safety. This study identified design and policy implications for hospital decision makers. In particular, it was shown that the technological capability of a hospital is a key consideration in determining the level of resources that are necessary to implement specific healthcare technologies within a hospital.


Synaptophysin immunoreactivity in the rat pituitary: alterations after 6-hydroxydopamine treatment.
Synaptophysin (SN) is a synaptic-vesicle-associated membrane protein whose presence is indicative of intact, functional synapses. This study examines the presence of SN in pituitary gland innervation after neurotoxin-induced denervation followed by reinnervation. Immunostaining of rat pituitary neurointermediate lobe tissue for SN reveals a pattern of dot-like densities in the intermediate lobe and intensely stained dispersed regions in the neural lobe of normal animals. In rats treated with 6-hydroxydopamine (6-OHDA), a catecholamine neurotoxin, by peripheral injection, there is a significant depletion of the SN immunostaining in the intermediate lobe, as well as a significant reduction of SN immunoreactivity in the neural lobe, in animals studied 1 wk after drug treatment, with computer analysis of the tissue sections. At 3 wk after 6-OHDA, there is a partial recovery of immunoreactivity for SN in the neural lobe in many tissue sections, and the intermediate lobe also contains only relatively sparse staining for the synaptic protein. Computer analysis revealed that at 3 wk after 6-OHDA, both lobes still had reduced SN immunoreactivity, but the difference in levels measured did not achieve statistical significance. These results contrast with the prior finding of significant recovery of immunoreactivity for GAP-43, a growth and regeneration-associated protein, in intermediate lobe innervation of rats treated with the same drug regimen. We suggest that 6-OHDA treatment damages synaptic vesicle integrity in both the intermediate and neural lobes of the pituitary, and that recovery is in progress, but not complete at 3 wk after the drug is administered.


Comfort and challenge: a thematic analysis of female clinicians' experiences of supervision.
The supervision of counsellors, counselling psychologists, and psychotherapists is generally perceived to be an invaluable component of training and practice. The present study analysed this process to explore the meanings of supervision and to consider implications for clinical practice and training. This study presents the accounts of 19 psychological therapists who experienced supervision while working at a London-based women's therapy centre. Demographic information was collected by questionnaire, and semi-structured interviews were conducted to explore aspects of the supervisory experience. The qualitative data were thematically analysed. Supervision has complex and paradoxical meanings, and it impacts on clinical counselling practice in multifarious ways. Dominant themes highlighted were the usefulness of supervision, specifically support, empowerment, and joining; fear of exposure in supervision versus gaining new information; the comfort versus the challenge of supervision; and supervision as a containing space. An experiential model of supervision was developed from the data. This model complements existing models in the field. It could be evaluated and used alongside existing models in various training and supervision contexts. Findings are discussed in relation to the literature on clinical supervision.


Effects of wortmannin and latrunculin A on slow endocytosis at the frog neuromuscular junction.
Phosphoinositides are key regulators of synaptic vesicle cycling and endocytic traffic; the actin cytoskeleton also seems to be involved in modulating these processes. We investigated the effects of perturbing phosphoinositide signalling and actin dynamics on vesicle cycling in frog motor nerve terminals, using fluorescence and electron microscopy, and electrophysiology. Antibody staining for beta-actin revealed that actin surrounds but does not overlap with synaptic vesicle clusters. Latrunculin A, which disrupts actin filaments by binding actin monomers, and wortmannin, an inhibitor of phosphatidyl inositol-3-kinase (PI3-kinase), each disrupted the pattern of presynaptic actin staining, but not vesicle clusters in resting terminals. Latrunculin A, but not wortmannin, also reduced vesicle mobilization and exocytosis. Both drugs inhibited the stimulation-induced uptake of the styryl dye FM1-43 and blocked vesicle reformation from internalized membrane objects after tetanic stimulation. These results are consistent with a role of PI3-kinase and the actin cytoskeleton in the slow pathway of vesicle endocytosis, used primarily by reserve pool vesicles.


Short-term effects of catheter-based renal denervation on cardiac sympathetic drive and cardiac baroreflex function in heart failure.
Sympathetic drive, especially to the heart, is elevated in heart failure and is strongly associated with poor outcome. The mechanisms causing the increased sympathetic drive to the heart remain poorly understood. Catheter-based renal denervation (RDN), which reduces blood pressure (BP) and sympathetic drive in hypertensive patients, is a potential treatment in heart failure. The aim of this study was to investigate the short-term effects of catheter-based RDN on BP, heart rate (HR) and cardiac sympathetic nerve activity (CSNA) and on baroreflex function in a conscious, large animal model of heart failure. Adult Merino ewes were paced into heart failure (ejection fraction<40%) and then instrumented to directly record CSNA. The resting levels and baroreflex control of CSNA and HR were measured before and 24h after bilateral renal (n=6) or sham (n=6) denervation. RDN was performed using the Symplicity Flex Catheter System® (Medtronic) using the same algorithm as in patients. Catheter-based RDN significantly reduced resting diastolic BP (P<0.01) and mean arterial blood pressure (P<0.05), but did not change resting HR or CSNA compared with sham denervation. Renal denervation reduced the BP at which CSNA was at 50% of maximum (BP50; P<0.005) compared with sham denervation. In an ovine model of heart failure, catheter-based RDN did not reduce resting CSNA in the short-term. There was, however, a lack of a reflex increase in CSNA in response to the fall in arterial pressure due to a leftward shift in the baroreflex control of CSNA, which may be due to denervation of renal efferent and/or afferent nerves.


Calorie restriction protects against age-related rat aorta sclerosis.
Many theories have been advanced to account for the ageing process, among which the free radical theory deserves much attention. Numerous studies have also shown an association between tissue fibrosis and oxidative stress. Of note, fibrosis may be considered a significant index of tissue ageing. Calorie restriction (CR) has been demonstrated to maintain many physiological processes in a youthful state until a very advanced age. However the anti-ageing mechanisms of CR are still not fully understood. We thus evaluated the effect of CR on oxidative damage and its relationship with fibrosis during ageing. We found a significant increase of both oxidative stress and fibrosis parameters in the aortae from aged vs. young rats. CR reversed both phenomena. CR also protected against the age-associated increase of Jun-N-terminal kinase and p-38 activities, involved in TGFbeta1 expression and signaling. On the contrary, extracellular regulated kinases did not show any age-related change. CR similarly reversed the age-related increase of AP-1 DNA binding activity and the AP-1-dependent increase of vimentin gene expression. In parallel, CR reversed the age-related morphological alterations of the aorta wall cell composition. These data further support the relationship between oxidative stress and fibrosis in different diseases and during ageing. The protection exerted by CR against fibrosclerosis might be due to a decrease of oxidative stress, with consequent decreased MAPK activity and down-regulation of AP-1 activation and of TGFbeta1 expression and signaling.


Attempt to quantify uterine involution using acoustic radiation force impulse before and after placental delivery.
The aim of the present study was to investigate whether baseline stiffness of the uterine corpus and cervix accurately estimated by acoustic radiation force impulse (AFRI) elastography changed after placental delivery. Eleven patients with normal vaginal delivery underwent ARFI elastography before, immediately after, and 1 and 2 h after placental delivery, and the shear wave velocity was measured to determine the stiffness. Each measurement was performed in triplicate to obtain a mean ± SD. The shear wave velocity of the uterine corpus before, immediately after, and 1 and 2 h after placental delivery was 1.81 ± 0.60, 3.04 ± 0.76, 3.12 ± 0.95, and 2.72 ± 0.81 m/s, respectively, and the shear wave velocity of the uterine cervix was 1.35 ± 0.45, 1.87 ± 0.57, 1.68 ± 0.59, and 1.70 ± 0.5 m/s, respectively. The stiffness of the uterine corpus significantly changed over time, although that of the uterine cervix was not significantly altered. The stiffness of the uterine corpus was significantly higher immediately after and 1 and 2 h after placental delivery as compared with that before placental delivery. The uterine corpus had a significantly higher stiffness than the uterine cervix at each of the four time points examined. ARFI elastography may be useful to assess uterine involution using the shear wave velocity.


Extracorporeal Membrane Oxygenation in the Therapy of Cardiogenic Shock: Results of the ECMO-CS Randomized Clinical Trial.
Veno-arterial extracorporeal membrane oxygenation (VA-ECMO) is increasingly being used for circulatory support in patients with cardiogenic shock, although the evidence supporting its use in this context remains insufficient. The ECMO-CS trial (Extracorporeal Membrane Oxygenation in the Therapy of Cardiogenic Shock) aimed to compare immediate implementation of VA-ECMO versus an initially conservative therapy (allowing downstream use of VA-ECMO) in patients with rapidly deteriorating or severe cardiogenic shock. This multicenter, randomized, investigator-initiated, academic clinical trial included patients with either rapidly deteriorating or severe cardiogenic shock. Patients were randomly assigned to immediate VA-ECMO or no immediate VA-ECMO. Other diagnostic and therapeutic procedures were performed as per current standards of care. In the early conservative group, VA-ECMO could be used downstream in case of worsening hemodynamic status. The primary end point was the composite of death from any cause, resuscitated circulatory arrest, and implementation of another mechanical circulatory support device at 30 days. A total of 122 patients were randomized; after excluding 5 patients because of the absence of informed consent, 117 subjects were included in the analysis, of whom 58 were randomized to immediate VA-ECMO and 59 to no immediate VA-ECMO. The composite primary end point occurred in 37 (63.8%) and 42 (71.2%) patients in the immediate VA-ECMO and the no early VA-ECMO groups, respectively (hazard ratio, 0.72 [95% CI, 0.46-1.12]; P=0.21). VA-ECMO was used in 23 (39%) of no early VA-ECMO patients. The 30-day incidence of resuscitated cardiac arrest (10.3.% versus 13.6%; risk difference, -3.2 [95% CI, -15.0 to 8.5]), all-cause mortality (50.0% versus 47.5%; risk difference, 2.5 [95% CI, -15.6 to 20.7]), serious adverse events (60.3% versus 61.0%; risk difference, -0.7 [95% CI, -18.4 to 17.0]), sepsis, pneumonia, stroke, leg ischemia, and bleeding was not statistically different between the immediate VA-ECMO and the no immediate VA-ECMO groups. Immediate implementation of VA-ECMO in patients with rapidly deteriorating or severe cardiogenic shock did not improve clinical outcomes compared with an early conservative strategy that permitted downstream use of VA-ECMO in case of worsening hemodynamic status. URL: https://www. gov; Unique identifier: NCT02301819.


Equivalence of mean intercept length and gradient fabric tensors - 3D study.
In this study the relationship between the mean intercept length (MIL) method - the current standard histomorphometric method of assessing structural anisotropy and an alternative method of the gray-level structure tensor (GST) is investigated. Both methods are applied to a set of 25 three-dimensional binary μCT images of trabecular bone. It is shown that there is a very strong correlation between the logarithms of the principal values of the MIL and the GST fabric tensors (Pearson's coefficient of correlation higher than 0.98) and between the logarithms of the invariants of the MIL and the GST fabric tensors (Pearson's coefficient of correlation higher than 0.999). There is also a good correlation between the degree of anisotropy calculated from the MIL and from the GST tensors (Pearson's coefficient of correlation equal to 0.90). The principal anisotropy directions of the MIL and the GST fabric tensors coincide at the 5% significance level. Additionally, the performance of both methods is tested, based on a set of artificial structures with prescribed orientations.


Prognostic significance of baseline reverse transcriptase-PCR for prostate-specific antigen in men with hormone-refractory prostate cancer treated with chemotherapy.
Methods accurately categorizing the diverse biology of prostate cancer are needed. A positive baseline reverse transcriptase-PCR for prostate-specific antigen (RT-PCR PSA) in the androgen-independent setting is an independent prognostic marker of survival. The objectives of the current study were to examine the prognostic implication of baseline RT-PCR PSA positivity during treatment with an active chemotherapeutic agent and explore whether an RT-PCR PSA "response" provides prognostic information. In a combined analysis of a phase I and a randomized phase II trial of BMS-247550 (an epothilone B analogue), 104 patients with hormone-refractory prostate cancer had whole blood samples collected at baseline, then with each cycle of therapy. RT-PCR PSA was assessed and related to time to progression (TTP). From 100 evaluable patients, 368 samples were received, of which 90.8% were evaluable for RT-PCR PSA status. Baseline RT-PCR PSA status was significantly associated with TTP (hazard ratio, 2.22; 95% confidence interval, 1.40-3.52). Twenty-six of 38 patients positive at first assessment had at least one follow-up RT-PCR PSA that was negative ("response"). In univariate analysis, RT-PCR PSA response was not significantly associated with TTP, but in multivariate analysis, RT-PCR PSA response was of borderline statistical significance in predicting TTP (hazard ratio, 0.41; 95% confidence interval, 0.16-1.01). These results provide further confirmation that baseline RT-PCR PSA is a statistically significant predictor of TTP in hormone-refractory prostate cancer. Moreover, this is the first report to suggest that RT-PCR PSA response during chemotherapy treatment may predict TTP.


Isolation and quantification of diarylheptanoids from European hornbeam (Carpinus betulus L.) and HPLC-ESI-MS/MS characterization of its antioxidative phenolics.
Detailed polyphenol profiling of European hornbeam (Carpinus betulus L.) bark, leaf, male and female catkin extracts was performed by high-performance liquid chromatography-diode array detection coupled to tandem mass spectrometry (HPLC-DAD-MS/MS). A total of 194 compounds were characterized and tentatively identified. Gallo- and ellagitannins dominated in the methanol extracts, while flavonol glycosides and methoxylated flavones prevailed in the ethyl acetate samples. In the quest for diarylheptanoids, twelve compounds were isolated by the combination of subsequent reversed-phase flash chromatographic and high-performance liquid chromatographic methods. The structural elucidation of the isolated components was performed by ultrahigh-performance liquid chromatography-Orbitrap mass spectrometry (UHPLC-Orbitrap-MS) as well as 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. Six known cyclic diarylheptanoids, together with a new compound were described in Carpinus betulus for the first time. The occurrence of a linear diarylheptanoid and a lignan has also been unprecedented in the genus Carpinus. Moreover, three known flavonol glycosides were isolated. Based on the identification of characteristic fragment ions, a new mass spectrometric fragmentation pathway for meta,meta-cyclophane-type diarylheptanoids was proposed. Quantities of the four major cyclic diarylheptanoids in European hornbeam were determined by a validated UHPLC-DAD method for the first time. The antioxidant properties of the extracts and the isolated compounds were assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Contribution of the individual constituents to the total radical scavenging activity of the samples was evaluated by an off-line DPPH-HPLC-DAD method. This allowed the identification of gallo- and ellagitannin derivatives as the constituents being primarily responsible for the antioxidant capacity of the extracts.


First molecular study in Lebanese patients with Cockayne syndrome and report of a novel mutation in ERCC8 gene.
Cockayne Syndrome (CS) is a rare autosomal recessive disorder characterized by neurological and sensorial impairment, dwarfism, microcephaly and photosensitivity. CS is caused by mutations in ERCC6 (CSB) or ERCC8 (CSA) genes. Three patients with CS were referred to the Medical Genetics Unit of Saint Joseph University. Sanger sequencing of both ERCC8 and ERCC6 genes was performed: ERCC8 was tested in all patients while ERCC6 in one of them. Sequencing led to the identification of three homozygous mutations, two in ERCC8 (p.Y322* and c.843 + 1G > C) and one in ERCC6 (p.R670W). All mutations were previously reported as pathogenic except for the c.843 + 1G > C splice site mutation in ERCC8 which is novel. Molecular diagnosis was established in all patients included in our study. A genotype-phenotype correlation is discussed and a link, between mutations and some specific religious communities in Lebanon, is suggested.


Orbital giant conjunctival epithelial primary cyst.
A 15-year-old male, with no previous traumatic o surgical ocular injury, presented with a right eye proptosis and inferior displacement which began 4 months earlier. Ultrasound and CT showed a cystic lesion of 3.5 × 2 cm in the superior orbit. Surgical resection was performed by transconjunctival orbitotomy. The histopathology examination showed a cyst with nonkeratinized epithelium, and without goblet cells. After 10 years of follow-up, there have been no new lesions. Conjunctival primary orbital cysts are very uncommon. They are usually of small-moderate size; giant cysts are exceptional. Diagnosis by imaging is essential to establish the differential diagnosis and surgical treatment.


INTELLANCE 2/EORTC 1410 randomized phase II study of Depatux-M alone and with temozolomide vs temozolomide or lomustine in recurrent EGFR amplified glioblastoma.
Depatuxizumab mafodotin (Depatux-M) is a tumor-specific antibody-drug conjugate consisting of an antibody (ABT-806) directed against activated epidermal growth factor receptor (EGFR) and the toxin monomethylauristatin-F. We investigated Depatux-M in combination with temozolomide or as a single agent in a randomized controlled phase II trial in recurrent EGFR amplified glioblastoma. Eligible were patients with centrally confirmed EGFR amplified glioblastoma at first recurrence after chemo-irradiation with temozolomide. Patients were randomized to either Depatux-M 1.25 mg/kg every 2 weeks intravenously, or this treatment combined with temozolomide 150-200 mg/m2 day 1-5 every 4 weeks, or either lomustine or temozolomide. The primary endpoint of the study was overall survival. Two hundred sixty patients were randomized. In the primary efficacy analysis with 199 events (median follow-up 15.0 mo), the hazard ratio (HR) for the combination arm compared with the control arm was 0.71 (95% CI = 0.50, 1.02; P = 0.062). The efficacy of Depatux-M monotherapy was comparable to that of the control arm (HR = 1.04, 95% CI = 0.73, 1.48; P = 0.83). The most frequent toxicity in Depatux-M treated patients was a reversible corneal epitheliopathy, occurring as grades 3-4 adverse events in 25-30% of patients. In the long-term follow-up analysis with median follow-up of 28.7 months, the HR for the comparison of the combination arm versus the control arm was 0.66 (95% CI = 0.48, 0.93). This trial suggests a possible role for the use of Depatux-M in combination with temozolomide in EGFR amplified recurrent glioblastoma, especially in patients relapsing well after the end of first-line adjuvant temozolomide treatment. (NCT02343406).


Mindfulness-enhanced internet-based cognitive behavioural therapy for anxiety and depression: Outcomes in routine care.
Clinical trials support the efficacy of mindfulness-enhanced, internet-based cognitive behavioural therapies (CBT) in reducing anxiety and depression symptom severity. However, it is unclear how effective these interventions are when delivered in routine clinical care settings. This study investigated the utilisation and treatment outcomes of an online mindfulness-enhanced CBT program in the general Australian community. Over a one-year study period, 2187 adults commenced the online mindfulness-enhanced CBT program. The program consisted of six sequential lessons to be completed within 90 days. Participants (mean age= 39 ± 14 years, 68 % female) completed measures of symptom severity for anxiety, depression, and psychological distress at pre-, mid- and post-treatment. Treatment effects were assessed via intention-to-treat linear mixed models with Hedges' g effect size calculations. From pre- to post-treatment, the mindfulness-enhanced CBT program was associated with medium and large effect sized reductions in generalised anxiety symptom severity (g = 0.80, 95%CI: 0.74-0.86), depression symptom severity (g = 0.74, 95%CI: 0.68-0.80), and psychological distress (g = 0.97, 95%CI: 0.90-1.03). Program adherence was modest with approximately 30 % of participants completing all six lessons of the program. This study is limited by its lack of a comparator condition, longer-term follow-up, and assessment of the reasons for participant non-adherence. The relative acceptability and effectiveness of mindfulness versus CBT components was not examined. This study supports the effectiveness of mindfulness-enhanced iCBT for symptoms of anxiety and depression when delivered in a routine care setting under the supervision of community health care practitioners.


Mechanical suppression of premature ventricular complexes during catheter ablation procedures.
Mechanical suppression of premature ventricular complexes (PVCs) is not a well-known observation. We retrospectively reviewed this phenomenon in the Ventricular Arrhythmia (VA) ablation procedures performed at Richard L. Roudebush Veterans Health Administration (VHA) center. Data from 40 consecutive patients who underwent VA ablation at VHA, Indianapolis, IN, with 44 VA was included in the study. Demographic and electrophysiological parameter data was collected. Overall the mean age of the population was 64 ± 11 years. The phenomenon of mechanical suppression was seen in 11 PVCs. The mean age was 59 ± 15 years in the group in which mechanical suppression was seen. Of the 11 cases, the site of earliest activation was seen in the coronary sinus in 8 and in the pulmonary artery in 3. In one case catheter ablation was not performed because of proximity to the left coronary artery system. However, sustained pressure at the site with earliest electrograms (-35 ms) and 95% pacematch resulted in long-term suppression of PVCs. In the cases in which mechanical suppression was seen, there was a statistically significant reduction in PVC burden compared to pre ablation PVC load (1.1% ± 1.50% (post ablation) versus 24.04% ± 13.07% (pre ablation) versus p < 0.05). In all the 11 cases the site of mechanical suppression was also the site with earliest electrograms. This case series illustrates phenomenon of mechanical suppression of PVCs as an indication for good site for successful ablation in unique veteran patient population.


Postmortem catecholamine levels in pericardial and cerebrospinal fluids with regard to the cause of death in medicolegal autopsy.
Previous studies have suggested that postmortem serum catecholamine levels reflect the magnitude of physical stress responses or toxic/hyperthermic neuronal dysfunction during the death process. The present study investigated postmortem adrenaline (Adr), noradrenaline (Nad), and dopamine (DA) levels in pericardial fluid (PCF) and cerebrospinal fluid (CSF) with regard to the cause of death, compared with right heart blood levels, in serial medicolegal autopsy cases with a postmortem time within 48 h (n=494). Correlations between PCF and CSF Adr levels, and those among right heart blood, PCF, and CSF DA levels were marked (r=0.66-0.83, p<0.0001), but were otherwise lower (r=0.22-0.44). With regard to the cause of death, Adr and Nad levels in PCF, CSF, and right heart blood mostly presented similar findings: these levels were generally high in injury, intoxication, and hyperthermia (heatstroke), but were low in hypothermia (cold exposure). DA levels at each site were higher in injury and intoxication. In addition, higher levels were detected for Nad levels in sharp instrument injury, as well as Adr, Nad, and DA in carbon monoxide intoxication at each site, and for CSF Nad in psychotropic drug intoxication. These findings suggest that characteristic elevations in Adr, Nad, and DA levels in PCF and CSF are involved in systemic responses to fatal stress and toxic neuronal dysfunction, reflecting the magnitude of such responses in individual cases.


Aplasia and displacement of the horizontal portion of the petrous carotid artery in mandibulofacial dysostosis.
The horizontal portion of the petrous carotid artery (HPCA) of mandibulofacial dysostosis (MFD) patients had anterior-inferior displacement or was aplasia at birth. Our findings indicate that most MFD patients have aplasia of the internal carotid artery. This information may be very important for safe reconstruction surgery in atresia. MFD rarely has malformations of the cardiovascular system such as those seen in Goldenhar syndrome. This study aimed to compare MFD patients and normal subjects with respect to the length and diameter of the HPCA. We radiographically studied 22 MFD patients (44 sides) using high-resolution computed tomography (HRCT), measuring the length and diameter of the HPCA and the angle between the right and left HPCA using computer software. Findings were compared with those in 86 ears with normal auricles (control group) using the nonparametric test. The HPCA in MFD patients averaged 23.2 mm in length and 4.8 mm in diameter. The angle between the right and left HPCA averaged 100.2 degrees. In MFD patients, on average, HPCA were 2.0 mm shorter (p=0.001) and 0.4 mm narrower (p=0.016) than in control subjects. Mean angle between right and left HPCA was 5.4 degrees more acute in MFD patients than in control subjects (p=0.026). Moreover, the genu of the vertical and horizontal petrous ICA in the MFD group was on average 3.2 mm more inferior than in controls (p<0.001).


[Stress and migraine].
The link between stress and migraine is complex. In its recent conception, stress is viewed as a transactional process between an individual and his-her environment in which the individual makes a response to an internal or external constraint. This paper reviews the evidence in favor of a relationship between stress and migraine. Many studies show that 50 to 80% of patients report stress as a precipitating factor for their migraine headaches. Many authors have suggested that acute stress can provoke biological modifications lowering the threshold of the individual's susceptibility to a migraine attack. It has also been shown that the incidence of migraine is higher when stress scores are higher in the previous year. This suggests that as well as being a precipitating factor of crisis, stress could also be a precipitating factor of illness in susceptible individuals. Moreover, stress can trigger migraine chronification. This has been shown in many retrospective studies and in one prospective study. Hyperalgesia and central sensitivity to pain induced by chronic stress can partly explain this phenomenon. Many retrospective studies also show that adverse events during childhood, like sexual and physical abuse, are more frequent in migraineurs than non-migraineurs. Nevertheless, there is no prospective study allowing considering a causal link between childhood abuse and migraine in adulthood. Another point that will be tackled is the comorbidity between stress related psychiatric disorders, such as post-traumatic stress disorder, and migraine. Here again, many studies conducted in huge samples from the general population are convincing. All that leads to propose stress management therapies to migraineurs. Randomized control trials and meta-analyses have shown that relaxation therapies, biofeedback and stress management cognitive behavioral therapies are effective in migraine prophylaxis, above all in children. The use of these therapies is of particular interest in association with pharmacological treatments in patients with frequent crises. However, the majority of the studies have poor methodological standards. Nevertheless, stress management therapies are proposed as prophylactic treatment in the French recommendations for migraine management.


Effect of the inoculation of plant growth-promoting rhizobacteria on the photosynthetic characteristics of Sambucus williamsii Hance container seedlings under drought stress.
Plant growth-promoting rhizobacteria (PGPR) are beneficial bacteria that survive within the range of plant rhizosphere and can promote plant growth. The effects of PGPR in promoting plant growth, activating soil nutrients, reducing fertilizer application, and improving the resistance of plant inducible system have been widely investigated. However, few studies have investigated PGPR as elicitors of tolerance to abiotic stresses, especially drought stress. In this study, the effects of Acinetobacter calcoaceticus X128 on the photosynthetic rate (Pn), stomatal conductance (Gs), intracellular CO2 concentration (Ci), and total chlorophyll content [Chl(a+b)] of Sambucus williamsii Hance seedling leaves under moderate drought stress and drought-rewatering conditions were determined. Compared with those of uninoculated seedlings, the average Pn values during the entire drought stress of inoculated seedlings increased by 12.99%. As the drought duration was lengthened, Ci of uninoculated leaves continued to increase after rapidly declining, whereas Gs continuously decreased. Furthermore, their photosynthetic properties were simultaneously restricted by stomatal and non-stomatal factors. After X128 inoculation, Ci and Gs of S. williamsii Hance leaves continued to decrease, and their photosynthetic properties were mainly restricted by stomatal factors. At the end of the drought stress, water stress reduced [Chl(a + b)] of S. williamsii Hance leaves by 13.49%. However, X128 inoculation decreased this deficit to only 7.39%. After water supply was recovered, Pn, Gs, and [Chl(a+b)] in uninoculated leaves were reduced by 14.23%, 12.02%, and 5.86%, respectively, relative to those under well-watered conditions. However, Ci increased by 6.48%. Compared with those of uninoculated seedlings, Pn, Gs, and [Chl(a+b)] in X128-inoculated seedlings were increased by 9.83%, 9.30%, and 6.85%, respectively. Therefore, the inoculation of X128 under arid environments can mitigate the reduction of chlorophyll, delay the restriction caused by non-stomatal factors to Pn in plant leaves under water stress, and can be more conducive to the recovery of photosynthetic functions of leaves after water supply is recovered.


Neoadjuvant chemotherapy for gastric cancer: update.
Neoadjuvant chemotherapy has recently received increasing attention in an attempt to increase the rate of complete tumor resections, combat systemic metastases, and prolong survival in patients with gastric cancer. The available data indicate that neoadjuvant chemotherapy is feasible and does not increase postoperative morbidity and mortality. Compared to the results that can today be obtained with primary resection and lymphadenectomy, however, preoperative chemotherapy has so far failed to show a clear increase in the rate of complete tumor removal in patients with resectable gastric cancer. In patients with locally advanced or unresectable gastric cancer, preoperative chemotherapy may cause substantial reduction in locoregional tumor mass and thus increase the resection rate. This finding appears to translate into a survival benefit for those who respond to chemotherapy and have subsequent complete tumor resection. Because of severe shortcomings in the study design of the published reports, definite conclusions cannot be drawn from the available studies. Randomized controlled prospective trials are therefore clearly warranted. Exact pretherapeutic tumor staging, standardized resection and lymphadenectomy techniques, diligent evaluation of the resected specimen, and close follow-up are essential when designing these trials to identify subgroups of patients who may benefit from neoadjuvant chemotherapy for gastric carcinoma.


A femoral artery cannula that allows distal blood flow.
A femoral artery cannula is used for certain types of circulatory support but can cause ischemia, especially during prolonged perfusion. This study tests the function of a femoral cannula designed to allow proximal and distal blood flow. Five pigs were used in the study. In each animal a distal-flow cannula was implanted in the femoral artery of one leg, and the same-sized standard cannula was implanted in the other. Blood was drained from the left atrium and delivered to the femoral artery through the distal-flow cannula or standard cannula by using a centrifugal pump. An ultrasonic flow probe and microspheres were used to quantify flow and perfusion distal to the cannula. Distal femoral flow and tissue perfusion were present in all animals (5/5) with the distal-flow cannula but only in 1 of 5 animals with the standard cannula (P < .048). Distal flow did not change with pump flow. Mean distal flow at each level of pump flow was higher with the distal-flow cannula (P < .05). Tissue perfusion was also higher with the distal-flow cannula (0.052 +/- 0.028 vs 0.010 +/- 0.022 mL x min(-1) x g(-1), P < .03). In the swine model the distal-flow cannula allowed greater and more consistent distal flow than the standard cannula. The use of a distal-flow cannula for circulatory support might reduce the risk of distal limb ischemia.


Associative morphine tolerance in the rat: examinations of compensatory responding and cross-tolerance with stress-induced analgesia.
The results of three experiments designed to examine the processes subserving associative tolerance to morphine's analgesic effects, as assessed by the tail-flick test, are reported. The experiments indicated that associative tolerance in male Holtzman rats was not subserved by conditioned compensatory responses but was cross-tolerant with an apparently nonopioid form of stress-induced analgesia (SIA). Experiment 1 showed that rats tested for morphine analgesia in a distinctive context that had been paired previously with morphine injections (5 mg/kg) were more tolerant than animals that had had this context explicitly unpaired with a series of morphine injections or animals that were drug-naive. There was no evidence of a conditioned compensatory response of hyperalgesia in animals given a saline injection in the presence of environmental stimuli that had been previously paired with morphine, even under test conditions designed to minimize the possibility of floor effects that might have obscured the detection of drug-compensatory hyperalgesia. Experiment 2 demonstrated that the footshock procedures used for stress induction produced an SIA that was not attenuated by naloxone (10 mg/kg). Experiment 3 replicated the associative tolerance phenomenon of Experiment 1 and again failed to find evidence of conditioned compensatory responses. It was found that animals exposed to footshock in the context that had been associated with morphine administration developed significantly less SIA than control animals. The relevance of these findings for associative models of drug tolerance is discussed.


Tungsten disulfide (WS2) nanosheet-based photoelectrochemical aptasensing of chloramphenicol.
A method is described for photoelectrochemical determination of chloramphenicol (CLOA). It is based on the use of (a) aptamers protected with photoactive WS2 nanosheets, and (b) DNase I-assisted target recycling. The DNA aptamer without label was employed for recognition of CLOA. In the absence of CLOA, the aptamer is adsorbed on the surface of WS2. This leads to a decrease of photocurrent due to the steric-hindrance effect of aptamer DNA. The adsorption of WS2 also protects the aptamer from digestion by DNase. In the presence of CLOA, the aptamer will be desorbed from the WS2 surface due to formation of an aptamer/CLOA conjugate. This results in an increased photocurrent due to a decreased amount of aptamer DNA on the electrode surface. The increase of photocurrent can be further improved by applying DNase triggered catalytic recycling of CLOA. Under optimal experimental conditions, the response is linear 10 pM - 10 nM CLOA concentration range, with a 3.6 pM lower detection limit (at 3σ). This method is acceptably selective, accurate and stable. It was applied to the determination of CLOA in spiked milk samples and gave satisfactory results. Graphical abstract A simple and sensitive photoelectrochemical apta-biosensor was fabricated for chloramphenicol detection. In this work, WS2 nanosheets were employed as photoactive material, and DNase I catalytic chloramphenicol recycling strategy was adopted to amplify the detection signal.


Impact of bone marrow mesenchymal stem cells on the submandibular gland structure of adult male albino rats exposed to cadmium chloride toxicity.
This study was carried out in submandibular salivary glands of rats to demonstrate the changes induced by cadmium intoxication and the possible prophylactic and therapeutic effects of bone marrow mesenchymal stem cells (BMSCs). Sixty-five rats were divided into five groups. Rats in Group I were controls while those of Group II received daily dose of 10 mg/kg cadmium for 24 days. Rats in Group III received single prophylactic dose of 1 × 106 BMSCs one week before cadmium administration. Rats of Group IV were concomitantly administered cadmium and BMSCs, while those of Group V received cadmium for 24 days and were then treated with single dose of 1 × 106 BMSCs. Rats of Groups I, II, III, and IV were euthanized at the end of the experiment while those of Group V were euthanized one week later. Salivary gland specimens were processed and stained with H&E and inducible nitric oxide synthase; other specimens were used to demonstrate metallothionein gene expression using RT-PCR, malondialdehyde and catalase enzymes were detected by ELISA. Groups III and IV had nearly comparable findings to Group I regarding histological pattern with normal gland features. Group III recorded a lower fold of change for metallothionein gene (1.14 ± 0.20), a lower malondialdehyde enzyme (21.67 ± 1.63 nmol/mg), and a higher catalase enzyme (66.33 ± 2.16 mmol/mg). Regarding all variables, significant differences were found between the different groups (P < 0.001). BMSCs have prophylactic and therapeutic effects against cadmium-induced cytotoxicity in rat salivary glands.


Medication-related osteonecrosis of the maxilla: Prognosis of oral surgery combined with endoscopic sinus surgery.
To evaluate the effect of functional endoscopic sinus surgery (FESS) on the resolution of maxillary medication-related osteonecrosis of the jaw (MRONJ). This study included 62 patients diagnosed with MRONJ in the maxillary posterior area with or without maxillary sinusitis (MS). All patients underwent oral surgery. The concomitant MS was evaluated, and if indicated, FESS was performed on the same operation. Follow-up included clinical and radiological examinations at 4 months postoperation. Of the 62 patients, 24 (38.7%) showed no evidence of MS and 38 (61.3%) showed MS. Advanced MRONJ (stage 3), which has bony destruction of the sinus floor and signs of MS according to its definition, was seen in 27 patients (43.5%). In stage 3 MRONJ, patients with combined treatment of oral surgery and FESS compared to those treated with oral surgery alone showed higher percentage of resolution (84.2%, 37.5%, respectively) at 4 months postoperatively, and the results were statistically significant. Simultaneous management of MRONJ and sinusitis with combined treatment of oral surgery and FESS for the treatment of maxillary MRONJ can be an effective method, especially for advanced cases, to address its associated lesions.


Extracardiac conduit as alternative repair for scimitar syndrome.
Scimitar syndrome has been repaired by different surgical procedures including intracardiac baffle technique, reimplantation of scimitar vein (SV) to the right atrium or the left atrium (LA). However, several anatomical variations such as short venous collector coursing deeply within the lung hilum with infradiaphragmatic drainage make the repair more challenging with conventional repair techniques. We present an alternative method for repair using a tube graft to connect the SV to the LA.


Total severity score and age predict long-term hospitalization in COVID-19 pneumonia.
Severe COVID-19 pneumonia implies increased oxygen demands and length of hospitalization (LOS). We aimed to assess a possible correlation between LOS and COVID-19 patients' clinical laboratory data of admission, including the total severity score (TSS) from chest computed tomography (CT). Data were assessed retrospectively at the General Hospital "Agios Pavlos" in Greece. Clinical laboratory data, TSS, and LOS were recorded. A total of 317 patients, 136 women and 181 men, with a mean age of 66.58 ± 16.02 years were studied. Significant comorbidities were hypertension (56.5%), dyslipidemia (33.8%), type 2 diabetes mellitus (22.7%), coronary heart disease (12.9%), underlying pulmonary disease (10.1%), and malignancy (4.4%). Inpatient time was related to age (p < 0.001), TSS (p < 0.001), time from symptom onset to hospitalization (p = 0.006), inhaled oxygen fraction (p < 0.001), fibrinogen (p = 0.024), d-dimers (p < 0.001), and C-reactive protein (p = 0.025), as well as a history of hypertension (p < 0.001) and type 2 diabetes mellitus (p < 0.008). The multivariate analysis showed a significant association of the LOS with age (p < 0.001) and TSS (p < 0.001) independent of the above-mentioned factors. Early identification of disease severity using the TSS and patients' age could be useful for inpatient resource allocation and for maintaining vigilance for those requiring long-term hospitalizations.


Study to establish visual acuity norms with Teller Acuity Cards II for infants from southern China.
To establish the norms of binocular and monocular acuity and interocular acuity differences for southern Chinese infants and compare these norms with the results for northern Chinese infants. A prospective, comparative, and noninterventional study was conducted from January to August 2018. Teller Acuity Cards II were used to determine the binocular and monocular acuity of infants. The tolerance intervals and limits with a stated proportion and probability were used to evaluate the norms of binocular and monocular acuity and interocular acuity differences. An unpaired t-test was used to compare the obtained norms with the reported northern Chinese norms. The tolerance intervals of binocular acuity (mean acuity of 3.73, 7.35, and 12.01 cpd, respectively, at 12, 24, and 36 months), monocular acuity (mean acuity of 2.88, 6.91, and 10.75 cpd, respectively, at 12, 24, and 36 months), and interocular acuity differences (mean difference of 0.92, 2.89, and 3.99 cpd, respectively, at 12, 24, and 36 months) were obtained, exhibiting an increasing trend with age. The binocular visual acuity norms of southern Chinese infants were significantly lower than those in northern China (4.37 vs. 6.9 cpd at 8 months and 7.35 vs. 26 cpd at 24 months) (P = 0.011). Northern and southern Chinese infants exhibited distinct acuity norms and visual development patterns. The establishment of population-specific visual acuity norms is necessary for current populations of infants from different regions.


Mechanisms Linking the Gut Microbiome and Glucose Metabolism.
This review details potential mechanisms linking gut dysbiosis to metabolic dysfunction, including lipopolysaccharide, bile acids, short chain fatty acids, gut hormones, and branched-chain amino acids.


Application of copper plates for frequency tuning of surface wired and wireless MRI coils.
This study shows how a copper plate could be used for frequency tuning of surface wired and wireless MRI coils. For this purpose, it is proposed to place the copper plate directly on their conducting circuit. This leads to increase in the resonance frequency of coils. The effect is most perceptible if the copper plate is comparable in size to the conducting circuit of radiofrequency (RF) coil. The experimental work was performed on a 7.05 T MR scanner using surface MRI coils operating on different resonance frequencies: 1H (300 MHz), 31P (121 MHz), 23Na (79 MHz), 13C (75 MHz). Application of copper plate for frequency tuning of wireless multi-turn multi-gap transmission line resonator (MTMG-TLR) was considered for the first time. The proposed method can be claimed if the nominal variable inductance or capacitance is not enough for tuning the resonance frequency of the MRI coil to a higher frequency range.


Binding of the 5-HT3 ligand, [3H]GR65630, to rat area postrema, vagus nerve and the brains of several species.
The radiolabelled derivative of the potent and selective 5-HT3 receptor antagonist, GR65630, has been shown to label 5-HT3 receptors in homogenates of rat entorhinal cortex. We now report on the specific binding of this radioligand in homogenates of 16 discrete brain areas from several species and nine areas of human brain. We have further characterised specific binding to homogenates of rat area postrema and vagus nerve. In all species examined (rat, mouse, rabbit and ferret) the highest level of specific [3H]GR65630 (0.2 nM) binding was found in homogenates of the area postrema (26-83 fmol/mg protein). Binding in forebrain areas varied with species, Rat and mouse brain had relatively high levels of binding in cortical areas. The ferret and rat had relatively high levels in forebrain dopamine-containing areas. Of the areas of human brain examined the amygdala showed the highest level of specific [3H]GR65630 (0.2 nM) binding (3.0 fmol/mg protein). Specific [3H]GR65630 (0.05-2 nM) binding to homogenates of rat vagus nerve and area postrema was saturable (Bmax: vagus nerve 89.1 fmol/mg protein, area postrema 44.4 fmol/mg protein) and of high affinity (Kd: vagus nerve 0.50 nM, area postrema 0.24 nM). Selective 5-HT3 receptor agonists and antagonists potently inhibited [3H]GR65630 (0.2 nM) binding to homogenates of rat area postrema and vagus nerve. There was a close correlation between drug affinities in each area.


Studying sensorimotor processing with physiology in behaving Drosophila.
The neural underpinnings of sensorimotor integration are best studied in the context of well-characterized behavior. A rich trove of Drosophila behavioral genetics research offers a variety of well-studied behaviors and candidate brain regions that can form the bases of such studies. The development of tools to perform in vivo physiology from the Drosophila brain has made it possible to monitor activity in defined neurons in response to sensory stimuli. More recently still, it has become possible to perform recordings from identified neurons in the brain of head-fixed flies during walking or flight behaviors. In this chapter, we discuss how experiments that simultaneously monitor behavior and physiology in Drosophila can be combined with other techniques to produce testable models of sensorimotor circuit function.


Optimal conditions for the generation of monoclonal antibodies using primary immunisation of mouse splenocytes in vitro under serum-free conditions.
We have further optimised the serum-free in vitro immunisation system described by Ossendorp et al. (J. Immunol. Methods 91, 257, 1986) for the generation of hybridomas secreting specific antibodies. Thyroglobulin and the hapten 2-phenyl-5-oxazolone coupled to chicken serum albumin were used as antigens. For an optimal outgrowth of antigen-specific B cells the presence of T cells, thymocyte-conditioned medium and antigen are required. The addition of supernatant from EL-4 cells (stimulated by phorbolmyristate acetate) inhibits the outgrowth of antigen-specific B cells. Using six-well plates with a surface area of 10 cm2 per well, an optimal IgM response was obtained when 10(7) splenocytes in a total volume of 2 ml/well were cultured for 3 days in the presence of antigen and thymocyte-conditioned medium. Increasing the concentration of cells whilst maintaining a constant surface area resulted in a decreased response.


Functional coculture of sympathetic neurons and cardiomyocytes derived from human-induced pluripotent stem cells.
Sympathetic neurons (SNs) capable of modulating the heart rate of murine cardiomyocytes (CMs) can be differentiated from human stem cells. The electrophysiological properties of human stem cell-derived SNs remain largely uncharacterized, and human neurocardiac cocultures remain to be established. Here, we have adapted previously published differentiation and coculture protocols to develop feeder-free SNs using human-induced pluripotent stem cells (hiPSCs). hiPSC-SNs were characterized in monoculture and coculture with hiPSC-CMs, using antibody labeling, enzyme-linked immunosorbent assay, and whole cell patch-clamp electrophysiology techniques. hiPSC-SNs stained positive for peripherin, tyrosine hydroxylase, and nicotinic acetylcholine receptors, the latter two colocalizing in somas and synaptic varicosities. hiPSC-SNs functionally matured in vitro and exhibited healthy resting membrane potentials (average = -61 ± 0.7 mV), secreted norepinephrine upon activation, and generated synaptic and action currents and inward and outward voltage-dependent currents. All hiPSC-SNs fired action potentials in response to current injection, local application of potassium, or spontaneously, followed by short-medium afterhyperpolarizations. hiPSC-SNs could successfully be maintained in coculture with hiPSC-CMs, and this induced further development of hiPSC-SN action potential kinetics. To test functional coupling between the neurons and cardiomyocytes, the hiPSC-CM beating response to nicotine-induced norepinephrine release was assessed. In neurocardiac cocultures, nicotine exposure significantly increased the hiPSC-CM spontaneous beating rate, but not in hiPSC-CM monocultures, supporting nicotinic neuronal hiPSC-SN stimulation directly influencing hiPSC-CM function. Our data show the development and characterization of electrophysiologically functional hiPSC-SNs capable of modulating the beating rate of hiPSC-CMs in vitro. These human cocultures provide a novel multicellular model to study neurocardiac modulation under physiological and pathological conditions.NEW & NOTEWORTHY We present data on a functional coculture between human-induced pluripotent stem cell-derived sympathetic neurons and cardiomyocytes. Moreover, this study adds significantly to the available data on the electrophysiological function of human-induced pluripotent stem cell-derived sympathetic neurons.


Renin profiling predicts neurohormonal response to sacubitril/valsartan.
Clinical trials and observational cohorts show that beneficial effects of sacubitril/valsartan are less strong in an appreciable proportion of patients with heart failure with reduced ejection fraction (HFrEF). Lower blood pressure and impaired renal function predict suboptimal sacubitril/valsartan titration and a less favourable response. Circulating renin encompasses neurohormonal activation, intravascular volume, and renal function. We hypothesized that renin may predict response to sacubitril/valsartan, assessed by changes in N-terminal fraction of pro-brain natriuretic peptide (NT-proBNP). We performed a prospective, open-label, real-life cohort study. The study population consisted of 80 consecutive HFrEF patients (age 66 ± 10 years, 83% men) planned to initiate sacubitril/valsartan. Clinical and biohumoral assessment, including a full neurohormonal panel, was performed at baseline and at 1, 3, and 6 month follow-up. Response to sacubitril/valsartan was defined as ≥30% reduction in NT-proBNP levels from baseline to 6 months. Patients in the lower renin tertile had higher blood pressure and plasma sodium concentration (all P < 0.05). At follow-up, 38 patients (48%) were classified as responders. Circulating renin was lower in the responder group compared with non-responders (19.8 mU/L, IQR 3.7-78.0 mU/L vs. 55.0 mU/L, IQR 16.4-483.1 mU/L; P = 0.004). After adjustment for age, renal function, and blood pressure, renin was independently associated to response to sacubitril/valsartan (P = 0.018). In our preliminary study, we show that circulating renin predicts reduction in NT-proBNP levels after sacubitril/valsartan initiation in HFrEF patients. Renin assessment might be useful to discriminate potential responders from the subgroup with a weaker expected benefit, thus needing a closer, tailored management strategy.


Long-term follow-up of neurodegenerative phenomenon in severe traumatic brain injury using MRI.
Traumatic brain injury (TBI) lesions are known to evolve over time, but the duration and consequences of cerebral remodelling are unclear. Degenerative mechanisms occurring in the chronic phase after TBI could constitute "tertiary" lesions related to the neurological outcome. The objective of this prospective study of severe TBI was to longitudinally evaluate the volume of white and grey matter structures and white matter integrity with 2 time-point multimodal MRI. Longitudinal MRI follow-up was obtained for 11 healthy controls (HCs) and 22 individuals with TBI (mean [SD] 60 [15] months after injury) along with neuropsychological assessments. TBI individuals were classified in the "favourable" recovery group (Glasgow Outcome Scale Extended [GOSE] 6-8) and "unfavourable" recovery group (GOSE 3-5) at 5 years. Variation in brain volumes (3D T1-weighted image) and white matter integrity (diffusion tensor imaging [DTI]) were quantitatively assessed over time and used to predict neurological outcome. TBI individuals showed a marked decrease in volumes of whole white matter (median -11.4% [interquartile range -5.8; -14.6]; p < 0.001) and deep grey nuclear structures (-17.1% [-10.6; -20.5]; p < 0.001). HCs did not show any significant change over the same time period. Median volumetric loss in several brain regions was higher with GOSE 3-5 than 6-8. These lesions were associated with lower fractional anisotropy and higher mean diffusivity at baseline. Volumetric variations were positively correlated with normalized fractional anisotropy and negatively with normalized mean diffusivity at baseline and follow-up. A computed predictive model with baseline DTI showed good accuracy to predict neurological outcome (area under the receiver operating characteristic curve 0.82 [95% confidence interval 0.81-0.83]) CONCLUSIONS: We characterised the striking atrophy of deep brain structures after severe TBI. DTI imaging in the subacute phase can predict the occurrence and localization of these tertiary lesions as well as long-term neurological outcome. ClinicalTrials.gov: NCT00577954. Registered on October 2006.


Electrophysiological evidences of mutual modulatory influences on the retinal activity of the crayfish Procambarus bouvieri (O).
Electroretinographic evoked potentials (ERGs) were recorded in dark adapted crayfish by the application of pulses of light (0.09 Cd/ft2) presented every 2.5 min. Heterolateral illumination (HI) for sixty min (0.06-0.3 Cd/ft2) induced up to 50% decrease in ERG after a latency of 12-25 min. ERG depression was proportional to the intensity of HI and also showed a circadian rhythm. During the alpha phase the ERG recovery started 3-10 min after HI was turned off. In contrast it started only after 10-20 min during the rho phase. The time course of the ERG depression, which was abolished in splitbrain animals, strongly suggests that a mutual modulatory influence, probably of neuroendocrine nature, is present in the crayfish visual system.


A Histologically Complete Response to Immunotherapy Using Pembrolizumab in a Patient with Giant Cell Carcinoma of the Lung: An Additional Report and Literature Review.
We previously reported a case of giant cell carcinoma in the lung, in which the use of antiprogrammed death 1 (PD-1) immunotherapy resulted in substantial tumor reduction. In the present study, we describe an additional clinical course. A 69-year-old woman was diagnosed with giant cell carcinoma of the lung in clinical stage IVB (T2bN0M1c, BRA). The tumor expressed programmed death ligand 1 (PD-L1) in a high proportion. The patient received stereotactic radiotherapy for two sites of small brain metastases, followed by immunotherapy using anti-PD-1 antibodies (pembrolizumab). The treatment exerted a substantial tumor reduction through four cycles. However, treatment was withdrawn due to renal dysfunction. The primary lung tumor continued to regress for an additional four months without any further therapy, resulting in a clinical stage of T1aN0M0. Salvage thoracic surgery was then performed to remove the tumor residue in the lung. Microscopic examination of the sample revealed no residual cancer. The patient was free from recurrence at 16 months post surgery. We then comprehensively reviewed lung sarcomatoid carcinoma cases in the literature, in which anti-PD-1 antibodies were implemented. The current literature and our own findings suggest sarcomatoid carcinomas express high levels of tumoral PD-L1 and can be effectively treated with anti-PD-1 antibodies.


O(p + 1) x O(p + 1)-Invariant hypersurfaces with zero scalar curvature in euclidean space.
We use equivariant geometry methods to study and classify zero scalar curvature O(p + 1) x O(p + 1)-invariant hypersurfaces in R2p+2 with p > 1.