Polychlorinated biphenyl profiles in ringed seals (Pusa Hispida) reveal historical contamination by a military radar station in Labrador, Canada.
Significant amounts of soil contaminated with polychlorinated biphenyls (PCBs) were discovered at a military radar station in Saglek Bay, Labrador, Canada, in 1996. Subsequent work showed elevated PCB concentrations in local marine sediments, in the benthic-associated food web, and in some ringed seals (Pusa hispida). The benthic-associated food web clearly reflected local PCB contamination, but the high PCB concentrations found in some ringed seals remained unexplained. In the present study, the authors assess the extent to which this local PCB source at Saglek Bay is contributing to the contamination of ringed seals in northern Labrador. Among 63 ringed seals sampled along the northern Labrador coast, 5 (8%) had PCB levels that were higher than recorded anywhere else in the Canadian Arctic. In addition, compared with seals exhibiting a long-range signal, 45% and 60% of subadults and adult males, respectively, exhibited heavier PCB congener profiles as characterized by principal components analysis, >1.6-fold higher PCB/organochlorine pesticides ratios, and higher PCB concentration-weighted average log octanol-water partition coefficient values, consistent with a local source. Despite the spatially confined nature of contaminated sediments in Saglek Bay, the influence of this PCB source is not inconsequential; PCB concentrations in locally contaminated adult males are 2-fold higher than concentrations in those exposed only to long-range PCB sources and exceed an established threshold of 1.3 mg/kg for adverse health effects in seals.


Development of a High-Throughput Screening Assay to Identify Inhibitors of the SARS-CoV-2 Guanine-N7-Methyltransferase Using RapidFire Mass Spectrometry.
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) represents a significant threat to human health. Despite its similarity to related coronaviruses, there are currently no specific treatments for COVID-19 infection, and therefore there is an urgent need to develop therapies for this and future coronavirus outbreaks. Formation of the cap at the 5' end of viral RNA has been shown to help coronaviruses evade host defenses. Nonstructural protein 14 (nsp14) is responsible for N7-methylation of the cap guanosine in coronaviruses. This enzyme is highly conserved among coronaviruses and is a bifunctional protein with both N7-methyltransferase and 3'-5' exonuclease activities that distinguish nsp14 from its human equivalent. Mutational analysis of SARS-CoV nsp14 highlighted its role in viral replication and translation efficiency of the viral genome. In this paper, we describe the characterization and development of a high-throughput assay for nsp14 utilizing RapidFire technology. The assay has been used to screen a library of 1771 Food and Drug Administration (FDA)-approved drugs. From this, we have validated nitazoxanide as a selective inhibitor of the methyltransferase activity of nsp14. Although modestly active, this compound could serve as a starting point for further optimization.


Electrospinning graphene quantum dots into a nanofibrous membrane for dual-purpose fluorescent and electrochemical biosensors.
Graphene quantum dots (GQDs) have become increasingly important for applications in energy materials, optical devices and biosensors. Here we report a facile technique to fabricate a nanofibrous membrane of GQDs by electrospinning water-soluble GQDs with polyvinyl alcohol (PVA) directly. The structure and fluorescence properties of the fabricated PVA/GQD nanofibrous membrane were investigated using scanning and transmission electron microscopy, and fluorescence microscopy. It was found that the electrospun PVA/GQD nanofibrous membrane has a three-dimensional structure with a high surface area to volume ratio, which is beneficial for the adsorption of electrolytes and the diffusion of reactants. For the first time, the created PVA/GQD nanofibrous membrane was utilized to fabricate dual-purpose fluorescent and electrochemical biosensors for highly sensitive determination of hydrogen peroxide (H2O2) and glucose. The experimental results indicated that the fluorescence intensity of the nanofibrous membrane decreased linearly with increasing H2O2 concentration, because the addition of H2O2 leads to fluorescence quenching of the GQDs, which endows the fabricated nanofibrous membrane with fluorescence activity. Besides, after binding glucose oxidase onto the created nanofibrous membrane, the fabricated nanofibrous membrane showed high sensitivity and selectivity for glucose detection. In addition, the PVA/GQD nanofibrous membrane can also be directly electrospun onto an electrode for electrochemical detection of H2O2. This novel nanofibrous membrane exhibits excellent catalytic performance and fluorescence activity, and therefore has potential applications for the highly stable, sensitive, and selective detection of H2O2 and glucose.


Protein disulfide isomerase is required for platelet-derived growth factor-induced vascular smooth muscle cell migration, Nox1 NADPH oxidase expression, and RhoGTPase activation.
Vascular Smooth Muscle Cell (VSMC) migration into vessel neointima is a therapeutic target for atherosclerosis and postinjury restenosis. Nox1 NADPH oxidase-derived oxidants synergize with growth factors to support VSMC migration. We previously described the interaction between NADPH oxidases and the endoplasmic reticulum redox chaperone protein disulfide isomerase (PDI) in many cell types. However, physiological implications, as well as mechanisms of such association, are yet unclear. We show here that platelet-derived growth factor (PDGF) promoted subcellular redistribution of PDI concomitant to Nox1-dependent reactive oxygen species production and that siRNA-mediated PDI silencing inhibited such reactive oxygen species production, while nearly totally suppressing the increase in Nox1 expression, with no change in Nox4. Furthermore, PDI silencing inhibited PDGF-induced VSMC migration assessed by distinct methods, whereas PDI overexpression increased spontaneous basal VSMC migration. To address possible mechanisms of PDI effects, we searched for PDI interactome by systems biology analysis of physical protein-protein interaction networks, which indicated convergence with small GTPases and their regulator RhoGDI. PDI silencing decreased PDGF-induced Rac1 and RhoA activities, without changing their expression. PDI co-immunoprecipitated with RhoGDI at base line, whereas such association was decreased after PDGF. Also, PDI co-immunoprecipitated with Rac1 and RhoA in a PDGF-independent way and displayed detectable spots of perinuclear co-localization with Rac1 and RhoGDI. Moreover, PDI silencing promoted strong cytoskeletal changes: disorganization of stress fibers, decreased number of focal adhesions, and reduced number of RhoGDI-containing vesicular recycling adhesion structures. Overall, these data suggest that PDI is required to support Nox1/redox and GTPase-dependent VSMC migration.


Prevalence and Severity of Psychiatric Disorders and Suicidal Behavior in Service Members and Veterans With and Without Traumatic Brain Injury: Systematic Review.
Assess prevalence and severity of posttraumatic stress disorder, suicidal behavior, and depressive, substance use, and anxiety disorders in US service members or Veterans with and without a deployment-related mild traumatic brain injury (TBI) (mTBI). Systematic review using multiple databases (January 2000 to October 2017). We included national or geographically diverse samples. Prevalence and severity of psychiatric conditions based on diagnostic codes, clinician assessments, and self-report measures with results stratified by sample type. We identified 11 studies on the basis of national samples and 22 studies on the basis of geographically diverse samples. Traumatic brain injury severity was not always ascertained or reported. In national studies, posttraumatic stress disorder, depressive disorder, substance use disorder, and anxiety disorder prevalence were higher in those with TBI than in those without. One national sample reported prevalence of suicide attempts. Across psychiatric conditions, strength of evidence ranged from insufficient to moderate. In geographically diverse samples, the pattern of findings was similar. National studies provided insufficient evidence on psychiatric condition severity; geographically diverse studies found greater severity of posttraumatic stress disorder symptoms with mixed results for symptoms of depressive or substance use disorders. Service members and Veterans with TBI history have higher prevalence and possibly severity of selected psychiatric conditions.


Stabilized Carbenium Ions as Latent, Z-type Ligands.
Controlling the reactivity of transition metals using secondary, σ-accepting ligands is an active area of investigation that is impacting molecular catalysis. Herein we describe the phosphine gold complexes [(o-Ph2 P(C6 H4 )Acr)AuCl]+ ([3]+ ; Acr=9-N-methylacridinium) and [(o-Ph2 P(C6 H4 )Xan)AuCl]+ ([4]+ ; Xan=9-xanthylium) where the electrophilic carbenium moiety is juxtaposed with the metal atom. While only weak interactions occur between the gold atom and the carbenium moiety of these complexes, the more Lewis acidic complex [4]+ readily reacts with chloride to afford a trivalent phosphine gold dichloride derivative (7) in which the metal atom is covalently bound to the former carbocationic center. This anion-induced AuI /AuIII oxidation is accompanied by a conversion of the Lewis acidic carbocationic center in [4]+ into an X-type ligand in 7. We conclude that the carbenium moiety of this complex acts as a latent Z-type ligand poised to increase the Lewis acidity of the gold center, a notion supported by the carbophilic reactivity of these complexes.


Circadian system functionality, hippocampal oxidative stress, and spatial memory in the APPswe/PS1dE9 transgenic model of Alzheimer disease: effects of melatonin or ramelteon.
Alzheimer disease (AD) is a neurodegenerative disorder that primarily causes β-amyloid accumulation in the brain, resulting in cognitive and behavioral deficits. AD patients, however, also suffer from severe circadian rhythm disruptions, and the underlying causes are still not fully known. Patients with AD show reduced systemic melatonin levels. This may contribute to their symptoms, since melatonin is an effective chronobiotic and antioxidant with neuroprotective properties. Here, the authors critically assessed the effects of long-term melatonin treatment on circadian system function, hippocampal oxidative stress, and spatial memory performance in the APPswe/PS1 double transgenic (Tg) mouse model of AD. To test if melatonin MT1/MT2 receptor activation, alone, was involved, the authors chronically treated some mice with the selective MT1/MT2 receptor agonist ramelteon. The results indicate that many of the circadian and behavioral parameters measured, including oxidative stress markers, were not significantly affected in these AD mice. During the day, though, Tg controls (Tg-CON) showed significantly higher mean activity and body temperature (BT) than wild-type (WT) mice. Overall, BT rhythm amplitude was significantly lower in Tg than in WT mice. Although melatonin treatment had no effect, ramelteon significantly reduced the amplitude of the BT rhythm in Tg mice. Towards the end of the experiment, Tg mice treated with ramelteon (Tg-RAM) showed significantly higher circadian rhythm fragmentation than Tg-CON and reduced circadian BT rhythm strength. The free-running period (τ) for the BT and locomotor activity (LA) rhythms of Tg-CON was <24 h. Whereas melatonin maintained τ at 24 h for BT and LA in both genotypes, ramelteon treatment had no effect. In the behavioral tests, the number of approaches and time spent exploring novel objects were significantly higher in Tg-CON than WT controls. Brain tissue analysis revealed significant reduction in hippocampal protein oxidation in Tg-MEL and Tg-RAM compared with Tg-CON animals. These results suggest that not all aspects of the circadian system are affected in the APPswe/PS1 mice. Therefore, care should be taken when extending the results obtained in Tg mice to develop new therapies in humans. This study also revealed the complexity in the therapeutic actions of melatonin and ramelteon in this mouse model of AD.


Altered renal alpha-adrenoceptor density induced by prolonged NaCl and DOCA administration in spontaneously hypertensive rats.
This study examines the effects of a 6-week exposure to 1% NaCl in tap water with and without desoxycorticosterone acetate (DOCA) on renal alpha adrenoceptors and systolic blood pressure (SBP) in spontaneously hypertensive adult rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. On normal sodium intake, SHR had higher renal alpha 1- (p less than 0.05) and alpha 2-adrenoceptor densities (p less than 0.001) and SBP (p less than 0.001) than WKY rats. Although WKY rats given either 1% NaCl, DOCA, or DOCA plus 1% NaCl developed hypertension after 6 weeks of treatment, only 1% NaCl administration for the same period produced an increase in the alpha 1-adrenoceptor density when compared to the control (p less than 0.001). In SHR, ingestion of 1% NaCl or DOCA plus 1% NaCl increased the already elevated alpha 2-adrenoceptor density (p less than 0.001) and SBP even more in this strain after 6 weeks of treatment. Equilibrium dissociation constants (Kd), however, were similar for both classes of receptors in experimental and control rats. The results of this study indicate that only the combination of 1% NaCl in tap water and DOCA administration is effective in accelerating hypertension in adult SHR. On the other hand, in this form of genetic rat hypertension, exaggerated sodium intake with or without DOCA administration could presumably be correlated with the increased renal alpha 2 adrenoceptors observed in these animals.


Isoniazid inhibition of mycolic acid synthesis by cell extracts of sensitive and resistant strains of Mycobacterium aurum.
Isonicotinic acid hydrazide (isoniazid; INH) inhibition of mycolic acid synthesis was studied by using cell extracts from both INH-sensitive and -resistant strains of Mycobacterium aurum. The cell extract of the INH-sensitive strain was inhibited by INH, while the preparation from the INH-resistant strain was not. This showed that the INH resistance of mycolic acid synthesis was not due to a difference in drug uptake or the level of peroxidase activity (similar in both extracts). As INH did not induce accumulation of any labeled intermediates, it is postulated that the drug acts either on production of labeled chain elongation precursors of mycolic acids or an early step of this elongation. The level of inhibition was not changed by addition of NAD or nicotinamide; thus, INH does not act on mycolic acid synthesis as an NAD antimetabolite. Benzoic or acetic acid hydrazides and known or postulated metabolites of INH (i.e., the corresponding acid, aldehyde, or alcohol) were not inhibitors of cell-free mycolic acid synthesis; the complete structure of INH was required, as already known for inhibition of mycobacterial culture growth. Extracts prepared from INH-treated cells showed reduced mycolic acid synthesis, and the inhibition level was not modified by either extensive dialysis or pyridoxal phosphate. This latter molecule efficiently antagonized INH action by reacting rapidly with INH, as shown by differential spectroscopy. Moreover, pyridoxal phosphate did not release inhibition of INH-treated extracts. It is proposed that INH may covalently react with an essential component of the mycolic acid synthesis system.


Pharmacokinetics of cefepime during continuous venovenous hemodiafiltration.
The objective of this study was to analyze the pharmacokinetics of cefepime, in six patients with acute renal failure related to septic shock, during continuous venovenous hemodiafiltration (CVVHD) (Hemospal AN 69S hemofilter; Hospal, Lyon, France). Six patients, mean age 65 +/- 4 years (range, 61 to 69), were included and each received 2 g of cefepime by intravenous infusion over a 30-min period every 12 h. Prefilter serum, dialysate outlet (DO), and ultrafiltrate samples were collected 0.47, 0.50, 0.57, 1, 3, 5, 7, and 12 h after the beginning of infusion. The time design of samples was optimized in accordance with the theory of D optimality. The cefepime concentrations were measured by high-performance liquid chromatography. The pharmacokinetics computation was carried out using P-PHARM software. Mean serum concentration peaks were 53 +/- 21.9 mg/liter (range, 13.0 to 68.9) one-half hour after the infusion. The mean elimination half-life was 8.11 +/- 2.22 h (range, 4.76 to 10.84). DO clearance was 66.57 +/- 30.14 ml/min (range, 38.66 to 119.87). The mean volume of distribution was 0.71 +/- 0.37 liters/kg of body weight. CVVHD was effective for cefepime elimination. In these subjects, the elimination half-life and DO clearance were almost constant. The results of this study suggested that a 2-g twice-daily infusion (usual dosage) was required for an effective concentration in this group of patients.


Improving Public Health Policy by Comparing the Public Response during the Start of COVID-19 and Monkeypox on Twitter in Germany: A Mixed Methods Study.
Little is known about monkeypox public concerns since its widespread emergence in many countries. Tweets in Germany were examined in the first three months of COVID-19 and monkeypox to examine concerns and issues raised by the public. Understanding views and positions of the public could help to shape future public health campaigns. Few qualitative studies reviewed large datasets, and the results provide the first instance of the public thinking comparing COVID-19 and monkeypox. We retrieved 15,936 tweets from Germany using query words related to both epidemics in the first three months of each one. A sequential explanatory mixed methods research joined a machine learning approach with thematic analysis using a novel rapid tweet analysis protocol. In COVID-19 tweets, there was the selfing construct or feeling part of the emerging narrative of the spread and response. In contrast, during monkeypox, the public considered othering after the fatigue of the COVID-19 response, or an impersonal feeling toward the disease. During monkeypox, coherence and reconceptualization of new and competing information produced a customer rather than a consumer/producer model. Public healthcare policy should reconsider a one-size-fits-all model during information campaigns and produce a strategic approach embedded within a customer model to educate the public about preventative measures and updates. A multidisciplinary approach could prevent and minimize mis/disinformation.


Effects of strain and age on prophylaxis and co-carcinogenesis of urethan-induced mouse lung adenomas by butylated hydroxytoluene.
A single i.p. injection of butylated hydroxytoluene (BHT) 6 h before a single urethan injection had varying effects on lung tumorigenesis in mice of different strains and ages. Strains exhibiting both high (A/J, SWR/J) and low (BALB/cByJ, 129/J, C57BL/6J) susceptibility to urethan tumorigenesis were tested in this study. BHT treatment decreased tumor multiplicity by an average of 32% in adult A/J mice but acted as a cocarcinogen by increasing tumor number 48% in adult SWR/J mice, 240% in adult C57BL/6J mice, 655% in adult 129/J mice, and 38% in 14-day-old A/J mice. The numbers of both alveolar type 2 cell-derived and bronchiolar Clara cell-derived lung adenomas were similarly affected by these BHT treatments. Such BHT pre-treatment had no effect on adenoma multiplicity in either young or adult BALB/cByJ mice. Multiplicity in young BALB/cByJ mice was also unaffected by chronic BHT administration following urethan, even though multiplicities increased several-fold with such treatment in adult mice of this strain. Since the mice showing co-carcinogenesis by BHT include strains which are both highly susceptible and relatively resistant to urethan induction of lung tumors, our results support a distinction between genes regulating susceptibility to urethan carcinogenesis and to tumor modulation by BHT.


Glycemic control and hunger during recovery from high- and moderate-intensity Nordic skiing in well-trained skiers.
The aim of this paper was to study the effects of Nordic ski training on post-exercise blood glucose (BG) regulation. Twenty-one (male N.=10; female N.=11) competitive college Nordic skiers (age 19.14±1.3 yrs; body fat percentage 14.9±6.2) completed two ski training conditions (high intensity [HI] and easy volume [MOD]) and one resting control (CON) condition on separate days. At rest on the control day and upon completion of ski training, a 75 g oral glucose tolerance test (OGTT) was administered. BG was measured prior to the OGTT and at 20-minute intervals for 80 minutes of passive rest. Hunger was assessed prior to the OGTT and at 40 and 80 minutes of rest via Visual Analog Scale. Data were analyzed using a two-way ANOVA with repeated measures and one-way ANOVA (SPSS v. 19). Significant time effects were present for BG. BG area under the curve (AUC) was significantly smaller (P<0.05) following HI (8825±149 arbitrary units [a.u.]) vs. CON (9493±168 a.u.), but not different from MOD (9087±227 a.u.). MOD AUC tended to be smaller than CON (P=0.064). Mean CON hunger was lower than MOD (P<0.05) and tended to be lower than HI (P=0.064). High-intensity Nordic skiing appears to enhance glycemic control when provided with an OGTT following acute exercise. The role that active muscle volume plays in this response should be investigated.


Renal and cardiac effects of antihypertensive treatment with ramipril vs metoprolol in autosomal dominant polycystic kidney disease.
Hypertension is a common complication in autosomal dominant polycystic kidney disease (ADPKD). This prospective randomized double-blind study was performed to compare the renal and cardiac effects of the ACE inhibitor ramipril and the beta-blocker metoprolol as first line therapy in ADPKD patients with hypertension. Forty-six hypertensive ADPKD patients were randomized to either ramipril (n = 23) or metoprolol (n = 23). Twenty-four hour (24-h) ambulatory blood pressure (BP), glomerular filtration rate (GFR) as calculated by the Cockcroft and Gault formula, urinary albumin excretion (albumin/creatinine ratio), and left ventricular mass index (LVMI) were established at baseline and at yearly intervals. The total follow-up was 3 years. Baseline characteristics were similar in both groups. Mean arterial pressure (MAP) decreased significantly in both the ramipril and the metoprolol group (-8 +/- 2 and -6 +/- 2 mmHg; both P < 0.01). There was a significant decline in renal function during follow-up which was similar in patients treated with ramipril or metoprolol (-2.5 +/- 0.7 vs -2.9 +/- 0.8 ml/min/year; P = NS). After the 3 years follow-up, no differences in GFR, LVMI and urinary albumin excretion were observed between the ramipril and the metoprolol group (80.7 +/- 10.7 vs 78.0 +/- 7.6 ml/min, 102.6 +/- 6.8 vs 100.3 +/- 5.4 g/m(2); and 42.6 +/- 12.3 vs 70.3 +/- 32.5 mg/g, respectively; all P = NS). A post-hoc analysis evaluating the effects of BP control, revealed that LVMI increased in patients with standard BP control while it remained stable in patients with rigorous BP control with a significant difference in LVMI between the groups after 3 years of follow-up (110.5 +/- 6.3 vs 90.9 +/- 4.7 g/m(2); P = 0.017). Also, by the end of the study albuminuria was lower in patients with rigorous vs standard BP control (23.5 +/- 6.7 vs 94.8 +/- 35.4 mg/g; P = 0.05). In our study population of hypertensive ADPKD patients, no differences in renal function, urinary albumin excretion and LVMI were detected between those treated with ramipril or metoprolol, respectively, during a 3 years follow-up. Rigorous BP control prevented an increase in LVMI and reduced urinary albumin excretion, suggesting a crucial role of BP control for slowing progression of cardiac and renal organ damage in ADPKD.


EZH2: not EZHY (easy) to deal.
Seminal discoveries have established that epigenetic modifications are important for driving tumor progression. Polycomb group (PcG) proteins are highly conserved epigenetic effectors that maintain, by posttranslational modification of histones, the silenced state of genes involved in critical biologic processes, including cellular development, stem cell plasticity, and tumor progression. PcG proteins are found in two multimeric protein complexes called Polycomb repressive complexes: PRC1 and PRC2. Enhancer of zeste homolog 2 (EZH2), catalytic core subunit of PRC2, epigenetically silences several tumor-suppressor genes by catalyzing the trimethylation of histone H3 at lysine 27, which serves as a docking site for DNA methyltransferases and histone deacetylases. Evidence suggests that overexpression of EZH2 is strongly associated with cancer progression and poor outcome in disparate cancers, including hematologic and epithelial malignancies. The regulatory circuit and molecular cues causing EZH2 deregulation vary in different cancer types. Therefore, this review provides a comprehensive overview on the oncogenic role of EZH2 during tumorigenesis and highlights the multifaceted role of EZH2, as either a transcriptional activator or repressor depending on the cellular context. Additional insight is provided on the recent understanding of the causes and consequences of EZH2 overexpression in specific cancer types. Finally, evidence is discussed on how EZH2 has emerged as a promising target in anticancer therapy and the prospects for targeting EZH2 without affecting global methylation status. Thus, a better understanding of the complex epigenetic regulatory network controlling EZH2 expression and target genes facilitates the design of novel therapeutic interventions.


Acute respiratory failure due to diffuse parenchymal lung diseases in a respiratory intensive care unit of North India.
Background: Acute respiratory failure (ARF) due to diffuse parenchymal lung diseases (DPLDs) is associated with high mortality. Whether ARF due to acute interstitial pneumonia (AIP), idiopathic pulmonary fibrosis (IPF) and non-IPF DPLDs behaves differently remains unclear. Methods: A retrospective analysis of consecutive DPLD subjects with ARF admitted to respiratory intensive care unit (RICU). The baseline clinical, demographic characteristics, cause of ARF and mortality were compared between the groups. Results: 145 (5.8% of RICU admission) subjects (mean [SD] age, 51.6 [14.7] years, 406% males) with DPLD-related ARF (17 AIP; 32 IPF; 96 non-IPF DPLD) were admitted. Common causes of ARF were acute exacerbation of the underlying DPLD (n=59, 40.4%) followed by infections (n=48, 37.5%). There was no difference in the peak, plateau and driving pressures across groups. The mortality rate was 45.5% (66/145) and was highest in AIP (82%) followed by IPF (59%) and non-IPF DPLD (34%). On multivariate logistic regression analysis, baseline APACHE II score, PaO2:FiO2 ratio, delta SOFA, and the use of invasive mechanical ventilation were independent predictors of mortality. The type of underlying DPLD however, did not affect survival. Conclusions: DPLD-related ARF is an uncommon cause of admission even in a RICU, and is associated with a high mortality. (Sarcoidosis Vasc Diffuse Lung Dis 2018; 35: 363-370).


Dioxin Exposure Impairs BMP-2-Mediated Spinal Fusion in a Rat Arthrodesis Model.
Cigarette smoking inhibits bone-healing and leads to increased rates of pseudarthrosis. However, the mechanisms behind these effects are controversial. Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin)--a cigarette smoke constituent and potent activator of the aryl hydrocarbon receptor (Ahr)--negatively impacts bone quality and osteoblast differentiation. We hypothesized that activation of the Ahr by dioxin would inhibit bone morphogenetic protein (BMP)-2-mediated spinal fusion in a rat arthrodesis model. Female Long-Evans rats were pretreated with dioxin or vehicle in six weekly doses, followed by bilateral posterior lumbar spinal fusion across the L4-L5 transverse processes using recombinant human BMP (rhBMP)-2. Treatments continued until sacrifice at four weeks postoperatively. A third group was treated with dioxin for six weeks, followed by a recovery period of four elimination half-lives to assess the reversible effects of dioxin exposure on spinal fusion capacity. Bone formation and fusion capacity were evaluated using fusion scoring, radiography, micro-computed tomography, and histologic analysis. Fusion scores for dioxin-treated and dioxin-recovery rats were significantly lower than those for controls. Although fusion rates were also significantly reduced in dioxin-treated animals relative to controls (50% versus 100%, respectively), rates were not significantly reduced in dioxin-recovery animals (80%). Dioxin treatment significantly inhibited spinal fusion in a rat arthrodesis model, and a prolonged cessation of dioxin exposure facilitated only a partial recovery of bone-healing capacity. This finding indicates that, although the effects of dioxin are persistent, an extended recovery from exposure could potentially restore bone regeneration in vivo. Development of a pharmacologic agent that reduces the adverse effects of cigarette smoke on bone-healing could prove useful to orthopaedic surgeons. Since dioxin and other similar cigarette smoke toxins exert their effects through Ahr pathway activation, the receptor represents a potential therapeutic target to improve spinal fusion rates in patients who smoke.


Strychnoxanthine, a monoterpene alkaloid of a novel skeletal type from Strychnos gossweileri.
Strychnoxanthine, an anhydronium base, was isolated from root barks of STRYCHNOS GOSSWEILERI, a few years ago. The structure of this new indole alkaloid is now established by spectral analysis, especially by (1)H NMR at 360 MHz.


Expression of genes responsible for the repair of mispaired bases of the DNA (MLH1) in invasive ductal breast carcinoma.
Breast cancer is a heterogeneous group of diseases determined and distinguished by cellular type, gene expression and clinical signs and symptoms. Identification of histological and biological markers is of great value in predicting the progression of tumor growth and anticipating the expected response to various treatment options. Due to a high degree of cell proliferation in breast tumors and high genetic instability of these tumors, as a consequence of defective DNA repair mechanisms, chemotherapy as a treatment option often renders very successful results. During our scientific research we wanted to determine the involvement of the genetic polymorphisms of DNA mismatch repair system (MLH1 gene) and the subsequent development of breast carcinoma. This study included 108 patients who were surgically treated for invasive breast cancer at the Department of Plastic, Reconstructive and Aesthetic Surgery, University Hospital "Dubrava". The expression of the MLH1 gene was determined by immunohistochemical methods. The results showed that 82.9% of tumor cells expressed the MLH1 gene. Analysis of survival rate for patients with invasive ductal breast cancer showed a statistically significant (p = 0.043) correlation with the expression of MLH1 genes. The overall five year survival rate of our patients was 78.7%. These results indicate that there is a possible involvement of MLH1 gene in the progression and development of breast cancer.


Research into the relationship between digital health literacy and healthy lifestyle behaviors: an intergenerational comparison.
Digital health literacy wields a pivotal role in individuals' health status in terms of seeking and choosing appropriate and accurate information, and useful services from a vast array of choices. This study is aimed at assessing the validity and reliability of the Turkish version of Digital Health Literacy Instrument (DHLI) and examining the relationship between DHL and the healthy lifestyle behaviors of participants from X, Y, and Z generations. In this study, to conduct a cross-sectional web-based survey, an online self-report questionnaire was built, and a convenience sample with a snowball approach was used. The study was conducted among 1,274 respondents aged between 18 and 64 years. Data collection tools consisted of the Personal Information Form, Lifestyle Behavior Scale II (HLBS II), and DHLI. Cultural validation and psychometric testing of DHLI, exploratory factor analysis, confirmatory factor analysis, Cronbach's alpha test, and bivariate and multivariate regression analysis were used for statistical analysis. In the study, the Turkish version of the DHLI tool consisting of six dimensions proved to be valid and reliable, and deemed appropriate for use across all age groups. The average digital health literacy of the respondents was sufficient, but the mean of healthy lifestyle behavior scores was moderate. There was a positive significant relationship between the total mean scores of DHLI and HLBS. Among the subdimensions of DHLI, while the highest mean scores were in DHLI Reliability, DHLI Privacy, and DHLI Search, DHLI Navigation and DHLI Relevance showed the lowest mean scores. DHLI Reliability, DHLI Relevance, and DHLI Adding Content were statistically significant predictors of health-related behaviors of the respondents. The most important feature and novelty of this study is that, although the DHLI scale has been widely translated for use in many countries, it has been translated and adapted to Turkish for the first time herein. The study offers crucial evidence about Generation X, Y, and Z's DHL level and its positive relationships with health-related behaviors. Therefore, the community and its partners should lead the way in empowering individuals to understand and use online information in an effective, secure, and health-promoting manner, along with governments.


Hospital design and staff perceptions: an exploratory analysis.
The movement in the health care design field to focus on "patient-centered care" and "healing environments" is often reflected in the health care facilities' incorporation of features meant to mitigate the stressful nature of serious illness. These features may range from a resource center (to allow patients and families to properly educate themselves about their illness) to providing space in rooms so family members can remain with patients, to more ambient features such as soft lighting, water features, or healing gardens. Research has shown that such features can in fact have positive effects for patients and may reduce stress and speed physical healing. What is not as well understood is how (if at all) the medical staff uses these types of features. Good health care cannot be administered without health care professionals, but factors such as job stress and burnout can impair both the physical and psychological health of the staff. A supportive organizational environment is imperative, but this article will set out to show that a supportive physical environment can be just as necessary in influencing how the staff views their workplace as well as how they might fare with regard to their own health. This article will examine a subset of results from a recent employee satisfaction survey given at a medical center. Exploratory analysis and interpretation will begin to shed light on whether or not building design can have an impact (positive or negative) on staff satisfaction. This can potentially act as a springboard to guide future empirical research in the area of health care design and its impact on staff satisfaction and well-being. By analyzing findings from a recent employee satisfaction survey, we can begin to understand how the hospital staff perceives the design of their building and begin to determine if these perceptions might impact health outcomes among the employees.


Reduced bacterial deposition and attachment by quorum-sensing inhibitor 4-nitro-pyridine-N-oxide: the role of physicochemical effects.
Surface-attached chemical groups that resist protein adhesion are commonly characterized as being hydrophilic, H-bond acceptors, non-H-bond donors, and electrically neutral. Quorum-sensing (QS) inhibitor 4-nitropyridine-N-oxide (4-NPO) that previously was found to decrease Pseudomonas aeruginosa biofilm formation possesses all of these characteristics, making this molecule an ideal antiadhesive compound. It was hypothesized that once 4-NPO adsorbs to either the solid surface or bacteria, resultant changes in the physical-chemical surface properties of the solid surface and bacteria will reduce the extent of bacterial adhesion. These physical-chemical effects take place prior to the commencement of already well-established QS biofilm-inhibition mechanisms. Bacterial adhesion experiments to silica conducted in quartz crystal microbalance with dissipation (QCM-D) and parallel plate flow cells demonstrated that 4-NPO reduces bacterial adhesion to silica-coated surfaces by the adsorption of 4-NPO to the silica surface as well to the outer membrane of both gram-negative P. aeruginosa PAO1 and gram-positive Staphylococcus aureus. 4-NPO effectively neutralizes both the bacterial and silica surface charge, and it is proposed that this neutralization of local surface charge heterogeneities by 4-NPO adsorption is the mechanism responsible for decelerating rates of bacterial deposition.


[Serum trace elements and myocardial infarction].
The authors report trace elements concentrations (zinc, nickel, selenium, copper, manganese and chromium) during myocardial infarction described in some publication. Serum zinc decreases during the first two days after acute myocardial infarction and returns to normal level in two weeks. Serum nickel increases during the first 36 hours and copper during the first four days. The different explanations of these disturbances are also exposed.


Iridotomy to slow progression of angle-closure glaucoma.
This is the protocol for a review and there is no abstract. The objectives are as follows: The primary objective is to assess the role of iridotomy-compared with observation-in the prevention of visual field loss for individuals who have primary angle closure or primary angle-closure glaucoma in at least one eye. We will also examine the role of iridotomy in the prevention of elevated intraocular pressure (IOP) in individuals with narrow angles (primary angle-closure suspect) in at least one eye.


Differential expression of cell cycle regulators in HCV-infection and related hepatocellular carcinoma.
To investigate cell cycle proteins in chronic hepatitis C virus infection in order to analyze their role in the process of hepatocyte transformation and to characterize their prognostic properties. Subjects of the current study included 50 cases of chronic hepatitis C (CHC) without cirrhosis, 30 cases of CHC with liver cirrhosis (LC), and 30 cases of hepatitis C-related hepatocellular carcinoma (HCC) admitted to the Department of Hepato-Gastroenterology, Theodor Bilharz Research Institute (TBRI), Giza, Egypt. Fifteen wedge liver biopsies, taken during laparoscopic cholecystectomy, were also included as normal controls. Laboratory investigations including urine and stool analysis, liver function tests and prothrombin concentration; serologic markers for viral hepatitis and ultrasonography were done for all cases of the study together with immunohistochemical analysis using primary antibodies against Cyclin D1, Cyclin E, p21, p27 and Rb/p105 proteins. Normal wedge liver biopsies didn't express Cyclin E or Rb/p105 immunostaining but show positive staining for Cyclin D1, p21 and p27. Cyclin D1 expressed nuclear staining that was sequentially increased from CHC to LC (P < 0.01) to HCC (P < 0.001) cases; meanwhile, Cyclin E revealed nuclear positivity only in the case of HCCs patients that was directly correlated to Rb/p105 immuno-reactivity. The expression of p21 and p27 was significantly increased in CHC and LC cases compared to normal controls and HCCs with no significant difference between well- and poorly-differentiated tumors. p21 showed only a nuclear pattern of staining, while, p27 presented with either cytoplasmic and/or nuclear reactivity in all studied cases. Correlation analysis revealed a direct relation between Cyclin D1 and p21 in CHC cases (P < 0.001), between Cyclin D1 and Cyclin E in HCCs (P < 0.01); however, an inverse relationship was detected between Cyclin D1 and p21 or p27 (P < 0.001) and between p21 and Rb/p105 (P < 0.05) in HCCs. Upregulation of Cyclin D1 in CHC plays a vital role in the development and differentiation of HCC; while, Cyclin E may be a useful marker formonitoring tumor behavior. p21 and p27 can be used as predictive markers for HCC. Furthermore, higher expression of Rb/p105 as well as inverse relation with p21 and histologic grades suggests its important role in hepatic carcinogenesis.


The RNA binding protein CsrA controls cyclic di-GMP metabolism by directly regulating the expression of GGDEF proteins.
The carbon storage regulator CsrA is an RNA binding protein that controls carbon metabolism, biofilm formation and motility in various eubacteria. Nevertheless, in Escherichia coli only five target mRNAs have been shown to be directly regulated by CsrA at the post-transcriptional level. Here we identified two new direct targets for CsrA, ycdT and ydeH, both of which encode proteins with GGDEF domains. A csrA mutation caused mRNA levels of ycdT and ydeH to increase more than 10-fold. RNA mobility shift assays confirmed the direct and specific binding of CsrA to the mRNA leaders of ydeH and ycdT. Overexpression of ycdT and ydeH resulted in a more than 20-fold increase in the cellular concentration of the second messenger cyclic di-GMP (c-di-GMP), implying that both proteins possess diguanylate cyclase activity. Phenotypic characterization revealed that both proteins are involved in the regulation of motility in a c-di-GMP-dependent manner. CsrA was also found to regulate the expression of five additional GGDEF/EAL proteins and a csrA mutation led to modestly increased cellular levels of c-di-GMP. All together, these data demonstrate a global role for CsrA in the regulation of c-di-GMP metabolism by regulating the expression of GGDEF proteins at the post-transcriptional level.


Ritodrine sulphation in the human liver and duodenal mucosa: interindividual variability.
The beta2-adrenoceptor agonist ritodrine has a bioavailability of 30% due to its presystemic metabolism and sulphation is an important metabolic route. The interindividual variability in the rate of ritodrine sulphation in 100 specimens of human liver and duodenum is reported. The final concentrations of ritodrine were 2 mM (duodenum) and 20 mM (liver). The mean estimates of ritodrine sulphation rate were 490 pmol x min(-1) x mg(-1) (duodenum) and 140 pmol x min(-1) x mg(-1) (liver). There was a 4-5-fold variation within +/- 2 SD units in the hepatic and duodenal rates of ritodrine sulphation. Statistical analysis revealed the presence of at least two subgroups of ritodrine sulphation. In the liver, 30% and 70% of the population fell into two subgroups with the mean estimates of ritodrine sulphation rate of 114 and 149 pmol x min(-1) x mg(-1), respectively (P < 0.05). In the duodenum, 25% and 75% of the population fell into two subgroups and the mean estimates of ritodrine sulphation rate were 332 and 538 pmol x min(-1) x mg(-1), respectively (P < 0.05). The rates of ritodrine and 4-nitrophenol sulphation correlated highly in the liver (r = 0.865; P < 0.001) and the rates of ritodrine and dopamine sulphation correlated highly (r = 0.914; P < 0.001) in the duodenum. In both tissues, the rates of ritodrine and (-)-salbutamol sulphation underwent a similar extent of variation and correlated highly. The intrinsic clearance of ritodrine sulphation was over one order of magnitude higher in the duodenum than in the liver suggesting that the duodenum is an important site of ritodrine sulphation.


Structure/function analysis of the murine CD95L promoter reveals the identification of a novel transcriptional repressor and functional CD28 response element.
CD28 costimulation, an important second signal for antigen-mediated T cell activation, is known to enhance expression of several genes important for the regulation of CD4+ T cell effector function including interleukin-2 and CD154. Previous studies demonstrate CD28-mediated enhancement of the transcription and expression of Fas ligand (CD95L) in T cell lines, suggesting a regulatory link between CD28 and CD95L expression. These results served as the basis for structure/function analysis of the CD95L promoter to elucidate the mechanism for CD28-mediated enhancement of CD95L. In this report, we describe a novel response element, located at -210 to -201 bp upstream of the transcription start site, that confers CD28 responsiveness to the CD95L gene. This response element is homologous to the CD28 response element (CD28RE) previously identified in the IL-2 promoter and bears structural similarities to a newly identified CD28RE in the CD154 promoter. We further demonstrate that CD28-mediated enhancement of promoter activity correlates with enhanced expression of CD95L mRNA, cell surface expression of CD95L protein, and increased apoptosis of CD95+ target cells. These results demonstrate a direct transcriptional regulatory role for CD28 in CD95L-mediated functional activity in CD4+ T cells. Mutational analysis of the CD95L promoter also reveals a novel transcriptional repressor element located approximately 60 bp 5' of the CD28RE. The repressor element bears sequence homology to an activator protein-1 element, constitutively binds c-Fos but not c-Jun, and is activation-independent.


Characterization of locomotor activity circadian rhythms in athymic nude mice.
The relation between circadian dysregulation and cancer incidence and progression has become a topic of major interest over the last decade. Also, circadian timing has gained attention regarding the use of chronopharmacology-based therapeutics. Given its lack of functional T lymphocytes, due to a failure in thymus development, mice carrying the Foxn1(Δ/Δ) mutation (nude mice) have been traditionally used in studies including implantation of xenogeneic tumors. Since the immune system is able to modulate the circadian clock, we investigated if there were alterations in the circadian system of the athymic mutant mice. General activity circadian rhythms in 2-4 month-old Foxn1(Δ/Δ) mice (from Swiss Webster background) and their corresponding wild type (WT) controls was recorded. The response of the circadian system to different manipulations (constant darkness, light pulses and shifts in the light-dark schedule) was analyzed. Free-running periods of athymic mice and their wild type counterpart were 23.86 ± 0.03 and 23.88 ± 0.05 hours, respectively. Both strains showed similar phase delays in response to 10 or 120 minutes light pulses applied in the early subjective night and did not differ in the number of c-Fos-expressing cells in the suprachiasmatic nuclei, after a light pulse at circadian time (CT) 15. Similarly, the two groups showed no significant difference in the time needed for resynchronization after 6-hour delays or advances in the light-dark schedule. The proportion of diurnal activity, phase-angle with the zeitgeber, subjective night duration and other activity patterns were similar between the groups. Since athymic Foxn1(Δ/Δ) mice presented no differences with the WT controls in the response of the circadian system to the experimental manipulations performed in this work, we conclude that they represent a good model in studies that combine xenograft implants with either alteration of the circadian schedules or chronopharmacological approaches to therapeutics.


Inhibition of neointimal smooth muscle accumulation after angioplasty by an antibody to PDGF.
Approximately 30 to 40 percent of atherosclerotic coronary arteries treated by angioplasty or by bypass surgery occlude as a result of restenosis. This restenosis is due principally to the accumulation of neointimal smooth muscle cells, which is also a prominent feature of the advanced lesions of atherosclerosis. The factors responsible for the accumulation of intimal smooth muscle cells have not been identified. Platelet-derived growth factor (PDGF) is a potent smooth muscle chemoattractant and mitogen. It is present in platelets and can be formed by endothelium, smooth muscle, and monocyte-derived macrophages. The development of an intimal lesion in the carotid artery of athymic nude rats induced by intraarterial balloon catheter deendothelialization was inhibited by a polyclonal antibody to PDGF. These data demonstrate that endogenous PDGF is involved in the accumulation of neointimal smooth muscle cells associated with balloon injury and may be involved in restenosis after angioplasty, and perhaps in atherogenesis as well.


A min-cut algorithm for the consistency problem in multiple sequence alignment.
Multiple sequence alignments can be constructed on the basis of pairwise local sequence similarities. This approach is rather flexible and can combine the advantages of global and local alignment methods. The restriction to pairwise alignments as building blocks, however, can lead to misalignments since weak homologies may be missed if only pairs of sequences are compared. Herein, we propose a graph-theoretical approach to find local multiple sequence similarities. Starting with pairwise alignments produced by DIALIGN, we use a min-cut algorithm to find potential (partial) alignment columns that we use to construct a final multiple alignment. On real and simulated benchmark data, our approach consistently outperforms the standard version of DIALIGN where local pairwise alignments are greedily incorporated into a multiple alignment. The prototype is freely available under GNU Public Licence from E.C.


Generation of Induced Pluripotent Stem Cell-Like Lines from Human Pancreatic Ductal Adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) has a dismal prognosis, mainly because the tumors are detected too late for effective treatment or for developing suitable therapeutics. Reprogramming cancer cells to pluripotency by induced pluripotent stem cell (iPSC) technology, which can be then programmed back to their original cellular state, allows for studying the dynamic events in the course of the disease progression. Thus, we applied iPSC technology to model early progression of PDAC. We showed that when an iPS-like cell line, designated 10-22, derived from human recurrent PDAC, was injected into immunodeficient mice, the cells consistently recapitulated preinvasive, pancreatic intraepithelial neoplasia (PanIN) to invasive stages of human PDAC. This model was recently validated by revealing a new biomarker that can classify early resectable PDAC patients from healthy subjects. The procedure to derive iPSCs from human PDAC is principally the same as the procedure to generate iPSCs from normal human fibroblast. However, the heterogeneous initial populations, different cellular states, and active memory of pancreatic epithelial cells challenge for making iPSC-like lines from human PDAC. Herein, we describe how to create and maintain iPSC-like line from human PDAC by lentiviral transduction of reprogramming factors.


Historical overview of the Department of Neurosurgery at Yonsei University College of Medicine in the Republic of Korea.
Originally founded in 1885, Gwanghyewon later became the Severance Hospital (named after philanthropist Louis Severance, who supported and funded the construction of a modern hospital) and Yonsei University College of Medicine. The Department of Neurosurgery at Severance Hospital was established in 1957, and its residency program began in 1961. Currently, the Department of Neurosurgery has 34 professors and 17 fellows; specialties include vascular, functional, pediatric, tumor, skull base, and spine neurosurgery. With its state-of-the-art neurosurgical facilities and services, the Department of Neurosurgery has developed into a department of excellence within the Yonsei University Health System. In this vignette, the authors present a historic overview of the Department of Neurosurgery.


Benzodiazepine receptor binding response to acute and chronic stress is increased in aging animals.
Benzodiazepine receptor binding in vivo, as determined by the uptake of the high-affinity specific benzodiazepine receptor ligand [3H]Ro15-1788, was examined following acute and chronic defeat stress in male mice aged 6 weeks, 7 months and 1 year. Specific uptake in 6-week-old mice was increased from control values only in the cerebellum following acute but not chronic stress. Specific uptake in the cortex and hypothalamus was unchanged from control values following both acute and chronic stress. Seven-month-old mice demonstrated an increased specific uptake in the cortex and cerebellum when measured immediately following both acute stress and the final session of chronic stress. This enhanced binding returned to baseline levels by 24 h after stress. One-year-old mice demonstrated no change in specific uptake when measured after acute stress, while binding was enhanced in all brain regions after the final session of chronic stress. This increased binding was still evident at 24 h after the cessation of chronic stress. Changes in benzodiazepine binding differ as a response to acute and chronic stress, and this response varies markedly with age.


Direct Observation of Amyloid β Behavior at Phospholipid Membrane Constructed on Gold Nanoparticles.
Amyloid β (Aβ) is a potential biomarker of Alzheimer's disease (AD), and its fibrillation behavior is of interest and value. In this study, the Aβ behaviors on phospholipid membranes were observed by Membrane Surface-Enhanced Raman Spectroscopy (MSERS) method. Phospholipid (PL) membranes, consisting of DMPC and DMPS with a molar ratio of 9:1, were fabricated on gold nanoparticles with diameter of 100 nm (Au@PL). Enhancement of the Raman intensity of Au@PL was increased by Aβ, with enhancement factor about 40. The H-bonding network was disturbed in presence of NaCl which covered Au@PL and made Au@PL away from one another. When Aβ was applied with Au@PL, the H-bonding network was disturbed just after mixing. As the reaction reaches to equilibrium, Aβ attracted neighbouring Au@PL and induced aggregation of Au@PL which blocked the aggregation prone site of Aβ to inhibit further fibrillation. Based on our method, the Aβ behaviors at lipid membrane surface can be directly observed via enhanced Raman signals.


WIF1 is a frequent target for epigenetic silencing in squamous cell carcinoma of the cervix.
Aberrant activation of the Wnt/β-catenin signaling axis is a prominent oncogenic mechanism in numerous cancers including cervical cancer. Wnt inhibitory factor-1 (WIF1) is a secreted protein that binds Wnt and antagonizes Wnt activity. While the WIF1 gene is characterized as a target for epigenetic silencing in some tumor types, WIF1 expression has not been examined in human cervical tissue and cervical cancer. Here, we show that WIF1 is unmethylated and its gene product is expressed in normal cervical epithelium and some cultured cervical tumor lines. In contrast, several cervical cancer lines contained dense CpG methylation within the WIF1 gene, and expression of both WIF1 transcript and protein was restored by culturing cells in the presence of the global DNA demethylating agent 5-aza-2'-deoxycytidine. Using single-molecule MAPit methylation footprinting, we observed differences in chromatin structure within the WIF1 promoter region between cell lines that express and those that do not express WIF1, consistent with transcriptional activity and repression, respectively. The WIF1 promoter was aberrantly methylated in ∼60% (10 of 17) high-grade highly undifferentiated squamous cell cervical tumors examined, whereas paired normal tissue showed significantly lower levels of CpG methylation. WIF1 protein was not detectable by immunohistochemistry in tumors with quantitatively high levels of WIF1 methylation. Of note, WIF1 protein was not detectable in two of the seven unmethylated cervical tumors examined, suggesting other mechanisms may contribute WIF1 repression. Our findings establish the WIF1 gene as a frequent target for epigenetic silencing in squamous cell carcinoma of the cervix.


Transferrin receptor overexpression enhances transferrin responsiveness and the metastatic growth of a rat mammary adenocarcinoma cell line.
We previously found that breast cancer cell transferrin receptor expression and proliferative response to transferrin often correlated with metastatic capability. To further explore this, we transfected mammary tumor cells with a cDNA coding for the transferrin receptor and examined the effects of its overexpression on various cellular properties. A human transferrin receptor expression plasmid was made by excising the cDNA for the receptor from pcDTR1 and ligating it into the multiple cloning site of pcDNAINeo. The resulting construct was transfected into the poorly metastatic rat MTLn2 line that expresses low endogenous levels of rat transferrin receptor, and transfection-induced receptor expression was ascertained using antibodies specific for the human protein. Approximately 50% of the initial geneticin-resistant transfected MTLn2 cells overexpressed human transferrin receptor protein. High expressors were further isolated by four sequential FACS sorts. The final cell population expressed approximately 3-7 times more cell surface transferrin receptor than did vector transfected controls. Both lines proliferated at the same rate in normal (medium plus 5% FBS) culture conditions. However, in serum-free conditions, the transferrin receptor overexpressor cells displayed a pronounced proliferative response to transferrin whereas the control line did not. When injected into the mammary fat pads of female nude mice, cells from both lines formed micrometastases to the lung that were specifically visualized by immunohistochemical staining of rat cytokeratin 17. This revealed that the transferrin receptor transfected line formed larger lesions of this nature than did cells from the vector transfected controls. When injected into the tail vein of female nude mice, the transferrin receptor overexpressors likewise formed gross lung metastases of remarkably greater size than did the vector only transfectants. Overexpression of cell surface human transferrin receptor on MTLn2 cells appeared to affect their in vitro growth response to transferrin and their ability to grow at a secondary site in vivo.


Role of different imaging modalities of vascular calcification in predicting outcomes in chronic kidney disease.
Vascular calcification (VC) is common among patients with chronic kidney disease (CKD). The severity of VC is associated with increased risk of cardiovascular events and mortality. Risk factors for VC include traditional cardiovascular risk factors as well as CKD-related risk factors such as increased calcium and phosphate load. VC is observed in arteries of all sizes from small arterioles to aorta, both in the intima and the media of arterial wall. Several imaging techniques have been utilized in the evaluation of the extent and the severity of VC. Plain radiographs are simple and readily available but with the limitation of decreased sensitivity and subjective and semi-quantitative quantification methods. Mammography, especially useful among women, offers a unique way to study breast arterial calcification, which is largely a medial-type calcification. Ultrasonography is suitable for calcification in superficial arteries. Analyses of wall thickness and lumen size are also possible. Computed tomography (CT) scan, the gold standard, is the most sensitive technique for evaluation of VC. CT scan of coronary artery calcification is not only useful for cardiovascular risk stratification but also offers an accurate and an objective analysis of the severity and progression.


[Working conditions of traditional birth attendants: some characteristics of rural home situations].
This is a study of qualitative and dialectical approach, which subject is to analyze the working conditions of the traditional birth attendants, in a reproductive perspective of work, and for this reason, economically undervalued. The analysis has been constructed having as central category the work, under marxist and feminist premises. Obtained results permit us to state these traditional birth attendants do their work in material and financial poor conditions, related to transportation, access, and still, working environment. The analysis of this practice has given us a possibility to present a discussion on some traits of the traditional birth attendant's work, which is characterized as a reproductive work, economically undervalued, autonomous and eminently feminine. It still configures as a social practice of popular health, legitimated by the community. In the rural domestic obstetrics practice, unequal relations were in evidence since men and women play different roles in the same occupation, being women's responsability the reproduction of the feminine role.


West nile virus infection in the Mesopotamia region, Syria border of Turkey.
We described the serological prevalence of West Nile Virus (WNV) antibodies among the human population in a historical and strategic region of Turkey. A serologic survey was conducted based on suspected cases in April, 2009, in the Mesopotamia region of Turkey, in the villages that were located alongside the Zergan River. All the sera were tested by enzyme-linked immunosorbent assay ELISA (Euroimmune™), and the positive samples were tested by immunofluorescent assay (IFA; Euroimmune™). As confirmation, neutralizing antibodies against WNV were tested by microneutralization assay (MNTA). In total, 307 individuals were included. The MNTA test was found to be positive among 52 individuals out of 307 (17%). In multivariate analysis, age >50 [odds ratio (OR)=5.2, confidence interval (CI) 2.76-9.97, p<0.001) and being in an occupational risk group (OR=2.02, CI 1.02-4.04, p=0.044) were found to be the risk factors for WNV seropositivity with the MNTA test. The physicians in the region should be aware of the risk of WNV infection and should be alerted to detect the clinical cases.


The water content of the stratum corneum in patients with atopic dermatitis. Measurement with the Corneometer CM 420.
The dry looking skin seen in many patients with atopic dermatitis reflects a defect in the epidermal barrier, the stratum corneum, as demonstrated by an increased transepidermal water loss (TEWL) and a decreased ability of the stratum corneum to bind water. The absolute amount of water within the stratum corneum is of importance both for barrier properties and for the clinical appearance of the skin. This water content was measured with a new instrument, the Corneometer CM 420, which takes advantage of the high dielectric constant of water. Forty patients with atopic dermatitis were studied--20 with dry skin and 20 with clinically normal skin on non-eczematous areas. The stratum corneum in dry skin was found to have a lower content of water than that in the clinically normal skin (p less than 0.01). Clinically normal skin in patients with atopic dermatitis did not differ significantly from normal control skin. An experiment was performed in vitro in an attempt to correlate the values obtained with the Corneometer to the absolute amount of water within the corneum.


Prognostic value of right ventricular free wall longitudinal strain in a large cohort of outpatients with left-side heart disease.
Right ventricular free wall longitudinal strain (RVFWLS) has been proposed as an accurate and sensitive measure of right ventricular function that could integrate other conventional parameters such as tricuspid annulus plane systolic excursion (TAPSE) and fractional area change (FAC%). The aim of the present study was to evaluate the relationship between RVFWLS and outcomes in stable asymptomatic outpatients with left-sided structural heart disease. We enrolled 458 asymptomatic patients with left-side heart diseases and any ejection fraction who were referred for echocardiography to two Italian centres. The composite endpoint of death for any cause and heart failure hospitalization was used as primary outcome of this analysis. After a mean follow-up of 5.4 ± 1.2 years, 145 patients (31%) reached the combined endpoint. Most of echocardiographic parameters were related to outcomes, including right ventricular functional parameters. Mean value of RVFWLS in our cohort was -21 ± 8% and it was significantly related to the combined endpoint and in multivariable Cox-regression model; when tested with other echocardiographic parameters that were significantly related to outcome at univariate analysis, RVFWLS maintained its independent association with outcome (hazard ratio 0.963, 95% confidence interval 0.948-0.978; P = 0.0001). The best cut-off value of RVFWLS to predict outcome was -22% (area under the curve 0.677; P < 0.001; sensitivity 70%; 65% specificity). RVFWLS may help clinicians to identify patients with left-sided structural heart disease at higher risk for first heart failure hospitalization and death for any cause.


High-quality, large-area MoSe2 and MoSe2/Bi2Se3 heterostructures on AlN(0001)/Si(111) substrates by molecular beam epitaxy.
Atomically-thin, inherently 2D semiconductors offer thickness scaling of nanoelectronic devices and excellent response to light for low-power versatile applications. Using small exfoliated flakes, advanced devices and integrated circuits have already been realized, showing great potential to impact nanoelectronics. Here, high-quality single-crystal MoSe2 is grown by molecular beam epitaxy on AlN(0001)/Si(111), showing the potential for scaling up growth to low-cost, large-area substrates for mass production. The MoSe2 layers are epitaxially aligned with the aluminum nitride (AlN) lattice, showing a uniform, smooth surface and interfaces with no reaction or intermixing, and with sufficiently high band offsets. High-quality single-layer MoSe2 is obtained, with a direct gap evidenced by angle-resolved photoemission spectroscopy and further confirmed by Raman and intense room temperature photoluminescence. The successful growth of high-quality MoSe2/Bi2Se3 multilayers on AlN shows promise for novel devices exploiting the non-trivial topological properties of Bi2Se3.


Circulating osteocalcin level is not associated with incident type 2 diabetes in middle-aged male subjects: mean 8.4-year retrospective follow-up study.
Recent human studies suggested that serum osteocalcin is associated with the cross-talk between bone and energy metabolism. The aim of this study was to determine whether serum osteocalcin level is independently associated with the development of type 2 diabetes. A retrospective cohort study was performed of 1,229 nondiabetic men, aged 25-60 years, who were recruited from the Health Promotion Center, Samsung Medical Center, between January 1997 and December 1997. They were followed regularly at the center on an out-patient basis and during hospitalization for a mean of 8.4 years, and the development of type 2 diabetes was determined. In the baseline analysis, BMI, body fat percentage, triglyceride, homeostasis model assessment of insulin resistance value, and plasminogen activator inhibitor-1 levels varied inversely with the osteocalcin tertiles, and serum high-density lipoprotein cholesterol levels increased with the osteocalcin tertiles. However, no differences were observed in fasting glucose and glycated hemoglobin levels across the osteocalcin tertiles. Incident type 2 diabetes occurred in 90 (7.3%) of the study subjects. In Cox proportional hazards models, however, no statistical differences in the development of type 2 diabetes across the osteocalcin tertiles were evident after adjustment of other risk factors for incident diabetes. Despite baseline associations with favorable metabolic parameters, the serum osteocalcin level was not associated with the development of type 2 diabetes in middle-aged males.


[Role of the diabetogenic factor in hypercoagulation of the blood].
I.v. injection of the diabetogenic factor induced disfunction of anticoagulation system in rats. The endogenous heparin concentration decreases in the blood and no proper defensive response to thrombosis provocation occurs. The diabetogenic factor seems to play an important role in the development of thrombosis and other complications in diabetes.


Shear Stress Increases V-H + -ATPase and Acidic Vesicle Number Density, and p-mTORC2 Activation in Prostate Cancer Cells.
Cells in the tumor microenvironment experience mechanical stresses, such as compression generated by uncontrolled cell growth within a tissue, increased substrate stiffness due to tumor cell extracellular matrix (ECM) remodeling, and leaky angiogenic vessels which involve low fluid shear stress. With our hypothesis that shear stress increases V-H + -ATPase number density in prostate cancer cells via activation of the mTORC1 and mTORC2 pathways, we demonstrated and quantified such a mechanism in prostate cancer cells. Moderately metastatic DU145 and highly metastatic PC3 prostate cancer cells were subjected to 0.05 dynes cm - 2 wall shear stress for 24 h, followed by immunocytochemistry and fluorescence measurements of β 1 integrin, endosome, lysosome, V-H + -ATPase proton pump, mTORC1, and p-mTORC2 antibodies. Post shear stress migration assays, and the effects of vacuolar proton pump inhibitor Bafilomycin A1 (60 nM, 24 h) as well as shear stress on the ICC fluorescence intensity of the proteins of interest were conducted with DU145 cells. Low fluid shear stress increases the fluorescence intensity of β 1 integrin, endosome, lysosome, V-H + -ATPase, mTORC1, and p-mTORC2 antibodies in PC3 and DU145 cells, and also increased cell migration. However, Bafilomycin A1 decreased fluorescence intensity of all of these proteins in DU145 cells exposed to shear stress, revealing that V-H + -ATPase controls the expression of these proteins. Prostate cancer cell mechanotransduction increases endosomes, lysosomes, and proton pumps-where increases have been associated with enhanced cancer aggressiveness. We also show that the prostate cancer cell's response to force promotes the cancer drivers mTORC1 and mTORC2.


[Monascus purpureus: a new fungus of allergenic relevance].
Anaphylactic reactions to food containing allergens in the consumption or preparation of food are well known. However, allergy in the preparation of sausages have rarely been described. In the present study a 26-year-old butcher was investigated who had a severe anaphylactic reaction developing sneezing, rhinitis, conjunctivitis, generalised pruritus, followed by widespread urticaria, Quincke's oedema and dyspnoe after starting to prepare sausages containing red yield rice. Red yield rice is produced from polished and washed rice by means of the fungus Monascus purpureus. It was the first time that Monascus purpureus could be shown as allergic agent by means of prick-to-prick test, Cellular Antigen Stimulation Test (CAST) and different other immunoblots.


Application of double focusing sector field ICP-MS for multielemental characterization of human hair and nails. Part I. Analytical methodology.
The capabilities of double focusing sector field inductively coupled plasma mass spectrometry (ICP-SMS) for the determination of 71 elements in hair and nails were studied. A microwave-assisted digestion procedure with nitric acid and hydrogen peroxide as well as direct sampling of the nails by laser ablation (LA) have been tested. Examples of spectral interferences are given and different correction procedures are discussed. Method detection limits below ng g(-1) were obtained for 39 elements investigated by using high-purity reagents and by taking special care to prevent contamination during preparation. However, these detection limits were insufficient for detection of some platinum group elements in the majority of the samples. The accuracy of the analytical procedure was estimated by analysis of the GBW07601 certified reference material as well as by participation in an interlaboratory comparison program. The reproducibility was assessed from replicate analysis (including sample preparation) and was found to be, as average values for all elements, 9-10% R.S.D. and 18-19% R.S.D. for hair and nails, respectively. Contribution from exogenous deposition was evaluated by analyzing samples before and after washing, as well as by studying spatial element distribution along hair and nails. It was found that even after sample washing, many elements are enriched in the surface of the nail.


PyGaze: an open-source, cross-platform toolbox for minimal-effort programming of eyetracking experiments.
The PyGaze toolbox is an open-source software package for Python, a high-level programming language. It is designed for creating eyetracking experiments in Python syntax with the least possible effort, and it offers programming ease and script readability without constraining functionality and flexibility. PyGaze can be used for visual and auditory stimulus presentation; for response collection via keyboard, mouse, joystick, and other external hardware; and for the online detection of eye movements using a custom algorithm. A wide range of eyetrackers of different brands (EyeLink, SMI, and Tobii systems) are supported. The novelty of PyGaze lies in providing an easy-to-use layer on top of the many different software libraries that are required for implementing eyetracking experiments. Essentially, PyGaze is a software bridge for eyetracking research.


The oxidative stress response in Enterococcus faecalis: relationship between H2O2 tolerance and H2O2 stress proteins.
The hydrogen peroxide (H2O2) stress response in Enterococcus faecalis ATCC19433 was investigated. A 2.4 mmol l-1 H2O2 pretreatment conferred protection against a lethal concentration (45 mmol l-1) of this agent. The relatively high concentrations of H2O2 used for adaptation and challenge treatments in Ent. faecalis emphasised the strong resistance towards oxidative stress in this species. Various stresses (NaCl, heat, ethanol, acidity and alkalinity) induced weak or strong H2O2 cross-protection. This paper describes the involvement of protein synthesis in the active response to lethal dose of H2O2, in addition to the impressive enhancement of synthesis of five H2O2 stress proteins. Combined results suggest that these proteins might play an important role in the H2O2 tolerance response.


[Effect of inducers of mixed-function microsomal oxidases on the immune response of mice induced by heterologous erythrocytes].
In experiments on male white mice immunized with sheep red blood cells the influence of various substances, differing in their chemical composition and biological activity, on the antibody titers and the number of rosette-forming cells in the spleen has been studied. These substances, acting as inductors of microsomal oxidases with mixed function (cytochrome P-450 and P-448), are phenobarbital, 3-methylcholanthrene, DDT, diphenin, rifampicin, benzodiazepin derivatives (diazepam and phenazepam). All monoxygenase activity inductors have been shown to exert a more or less pronounced immunosuppressive effect. Thus, reciprocal relations between the induction of monoxygenases and the induction of immune response have been established. The results obtained in this research are discussed from the viewpoint of the new hypothesis stating that immunity is one of the functions of the general system metabolizing chemical compounds in the body.


The ZmbZIP22 Transcription Factor Regulates 27-kD γ-Zein Gene Transcription during Maize Endosperm Development.
Zeins are the most abundant storage proteins in maize (Zea mays) kernels, thereby affecting the nutritional quality and texture of this crop. 27-kD γ-zein is highly expressed and plays a crucial role in protein body formation. Several transcription factors (TFs) (O2, PBF1, OHP1, and OHP2) regulate the expression of the 27-kD γ-zein gene, but the complexity of its transcriptional regulation is not fully understood. Here, using probe affinity purification and mass spectrometry analysis, we identified ZmbZIP22, a TF that binds to the 27-kD γ-zein promoter. ZmbZIP22 is a bZIP-type TF that is specifically expressed in endosperm. ZmbZIP22 bound directly to the ACAGCTCA box in the 27-kD γ-zein promoter and activated its expression in wild tobacco (Nicotiana benthamiana) cells. 27-kD γ-zein gene expression was significantly reduced in CRISPR/Cas9-generated zmbzip22 mutants. ChIP-seq (chromatin immunoprecipitation coupled to high-throughput sequencing) confirmed that ZmbZIP22 binds to the 27-kD γ-zein promoter in vivo and identified additional direct targets of ZmbZIP22. ZmbZIP22 can interact with PBF1, OHP1, and OHP2, but not O2. Transactivation assays using various combinations of these TFs revealed multiple interaction modes for the transcriptional activity of the 27-kD γ-zein promoter. Therefore, ZmbZIP22 regulates 27-kD γ-zein gene expression together with other known TFs.


Development of the human medullary arcuate nucleus from mid-gestation to the perinatal period: A morphometric study.
Development of the human medullary arcuate nucleus (AN) has not been sufficiently investigated. The present study provides morphometric data by examining the brains from preterm and perinatal infants. Nine brains were obtained from infants aged 21-43 postmenstrual weeks (PW). Serial celloidin sections were cut and stained using the Klüver-Barrera method. After microscopic observations, morphometric parameters [AN volume, numerical density (Nv) and total number (Nt) of neurons, and neuronal profile area (PA)] were analyzed. The AN was found as a pair of neuronal masses on the ventral medullary surface at 21 PW. Caudally, it was ventrolateral to the pyramidal tract (PT), and rostrally, medial to the PT. In the middle, it was diminished in size or interrupted. The AN neurons were gradually enlarged with age, showing multiplicity in size and shape. The following findings had a marked asymmetry and individual variability: (1) complete or partial inclusion of the AN in the PT; (2) connection between the rostral AN and the pontine nuclei; (3) coexistence of pyknotic neurons. The AN volume increased exponentially with age, while the Nv decreased exponentially. The Nt changed along two phases (decrease-increase) after mid-gestation. The mean PA increased linearly with age. Asymmetry and/or individual variability were demonstrated in the AN volume, Nt, and mean PA. Asymmetry and individual variability in the AN morphology are present in fetal period. The AN may undergo neuron death and neuroblasts production in tandem after mid-gestation.


The significance of protein C antigen in acute and chronic liver biliary disease.
Protein C, a naturally occurring inhibitor of blood coagulation, was measured immunologically in 160 patients with acute and chronic liver and biliary disease. In 31 patients with acute viral hepatitis serially studied from admission to discharge from hospital, protein C antigen (PC:Ag) was low on admission in a high proportion of cases (61%) but became normal in 90% of them after two weeks at a time when the prothrombin time was still prolonged in 46% of the cases. PC:Ag was also low in 25 cirrhotic patients and in 20 patients with chronic active hepatitis. In chronic hepatitis and cirrhosis, PC:Ag levels significantly correlated with indexes of liver synthetic function. In primary biliary cirrhosis (n:40), PC:Ag was low in patients with advanced disease (stages III-IV) but high in the early phases, when cholestasis was not yet accompanied by impaired protein synthesis. PC:Ag was also very high in 20 patients with large bile duct obstruction and highly correlated with indexes of cholestasis. The authors' findings indicate that PC:Ag is reduced in liver disease proportionally to the impairment of the liver synthetic function and that its normalization after acute hepatitis might represent an early marker of recovery of this function.


Additivity of near work-induced transient myopia and its decay characteristics in different refractive groups.
To determine the additive effect, if any, of NITM after 1 and 2 hours of reading in different refractive groups. Fifteen early-onset myopes (EOMs), 14 late-onset myopes (LOMs), and 15 emmetropes (EMMs), as well as progressive myopes (PMs) and stable myopes (SMs), were tested. Subjects read binocularly for 2 hours at a distance of 35 to 40 cm. Distance refractive state of the right eye was assessed every 2 seconds for 30 seconds after the first hour of reading, and then every 2 seconds for 120 seconds after the second hour of reading. NITM was calculated as the difference in posttask distance refractive state compared with the pretask distance refractive state after each hour. Initial NITM values (mean +/- SE) recorded at the end of the near work tasks were 0.22 +/- 0.03 D and 0.29 +/- 0.03 D for the EOMs, 0.14 +/- 0.02 D and 0.20 +/- 0.03 D for the LOMs, 0.14 +/- 0.02 D and 0.15 +/- 0.02 D for the EMMs, 0.20 +/- 0.03 D and 0.27 +/- 0.03 D for the PMs, and 0.09 +/- 0.04 D and 0.20 +/- 0.05 D for the SMs, after the first and second hours of reading, respectively. After the second hour, only in the EOMs and LOMs was NITM significantly greater than that found after the first hour. Seventy percent of the myopes (EOMs and LOMs) but only 47% of the EMMs exhibited increased NITM in the second hour compared with the first hour. Only EOMs exhibited longer decay duration after the second hour of reading. NITM was increased in progressive myopes (PMs), but not in stable myopes (SMs), after the first hour of reading only. Within the PMs, NITM was increased after the second hour compared with the first hour. The time constant for decay was greater in the PMs than in the SMs. Lastly, many myopes (up to 46%) did not experience decay to baseline after the near task over the 120-second posttask period. EOMs and LOMs demonstrated larger NITM than the EMMs and exhibited NITM additivity, but the EOMs also exhibited prolonged decay of NITM compared with the EMMs and LOMs. PMs, but not SMs, exhibited additivity of NITM. These findings may be attributed to impaired sympathetic function in the subjects with myopia. It is speculated that with repeated cycles of near work, residual NITM may contribute to the progression of permanent myopia.


Costs and benefits of high mutation rates: adaptive evolution of bacteria in the mouse gut.
We have shown that bacterial mutation rates change during the experimental colonization of the mouse gut. A high mutation rate was initially beneficial because it allowed faster adaptation, but this benefit disappeared once adaptation was achieved. Mutator bacteria accumulated mutations that, although neutral in the mouse gut, are often deleterious in secondary environments. Consistently, the competitiveness of mutator bacteria is reduced during transmission to and re-colonization of similar hosts. The short-term advantages and long-term disadvantages of mutator bacteria could account for their frequency in nature.


Localized inhibition of platelets and platelet derived growth factor by a matrix targeted glycan mimetic significantly attenuates liver fibrosis.
New therapeutic strategies are needed for the growing unmet clinical needs in liver disease and fibrosis. Platelet activation and PDGF activity are recognized as important therapeutic targets; however, no therapeutic approach has yet addressed these two upstream drivers of liver fibrosis. We therefore designed a matrix-targeting glycan therapeutic, SBR-294, to inhibit collagen-mediated platelet activation while also inhibiting PDGF activity. Herein we describe the synthesis and characterization of SBR-294 and demonstrate its potential therapeutic benefits in vitro and in vivo. In vitro SBR-294 was found to bind collagen (EC50 = 23 nM), thereby inhibiting platelet-collagen engagement (IC50 = 60 nM). Additionally, SBR-294 was found to bind all PDGF homodimeric isoforms and to inhibit PDGF-BB mediated hepatic stellate cell activation and proliferation. Translating these mechanisms in vivo, SBR-294 reduced fibrosis by up to 54% in the CCl4 mouse model (p = 0.0004), as measured by Sirius red histological analysis. Additional fibrosis measurements were also supportive of the therapeutic benefit in this model. These results support the therapeutic benefit of platelet and PDGF antagonism and warrant further investigation of SBR-294 as a potential treatment for liver fibrosis.


An Operational Approach for Optimizing Transcranial Direct Current Stimulation.
Transcranial direct current stimulation (tDCS) is a non-invasive brain stimulation technique that has been utilized for treating brain disorders and improving cognitive function. In order to achieve targeted tDCS, many optimization methods of montages and electric currents have been proposed. However, these methods have some limitations. Most of them were proposed for single-objective optimization (focality or intensity) and have no constrain with the number of electrodes (Most devices only have less than 8 electrodes currently). In this study, we proposed an operational optimization approach for well-targeted tDCS, which aims to optimize for two objectives of electric field (EF) intensity and focality with constraints on the number of electrodes. Compared with traditional tDCS in our cohort (10 subjects), our method significantly improves the EF focality. When compared to commonly used 4×1 high-definition tDCS (HD-tDCS), our method can achieve higher EF intensity in the target region with less than 8 electrodes. Our method can balance the two objectives of EF and shorten optimization time, which is convenient for practical application.


Reactivation of apo horse liver alcohol dehydrogenase with the monovalent metal ion Ag(i).
Each subunit of the liver alcohol dehydrogenase dimer contains one catalytic and one structural Zn(II) atom. Enzyme with the catalytic metal atoms selectively removed is inactive but can be partly reactivated in the presence of Ag(I) ions. Reactivation results from Ag(I) ions entering the empty metal-binding site in the catalytic center. The specific activity of this silver enzyme reached 24% of the native enzyme. Atomic absorption analysis gave equal amounts of Ag(I) and Zn(II), corresponding to one mole of each metal per monomer. Metal-directed affinity labelling using bromo-imidazolyl propionate showed that the properties of the silver-reactivated enzyme were distinct from those of the native enzyme.


[Effect of staging treatment of Tongyuan acupuncture on pregnancy outcome in patients with recurrent implantation failure of thin endometrium type].
To compare the effect of Tongyuan acupuncture combined with medication and medication alone on pregnancy outcome in patients with recurrent implantation failure (RIF) of thin endometrium type. A total of 74 patients with RIF of thin endometrium type undergoing freeze-thaw embryo transfer were randomly divided into an observation group (37 cases) and a control group (37 cases). The patients in the control group were treated with freeze-thaw embryo transfer in hormone replacement cycle, and the estradiol valerate tablets were taken orally from the fifth day of menstruation, 2 mg per day. On the basis of the control group, the observation group was additionally treated with Tongyuan acupuncture at Baihui (GV 20), Dazhui (GV 14), Qihai (CV 6), Guanyuan (CV 4), etc., combined with other acupoints based on syndrome differentiation and menstrual stage, once every other day. Both groups were treated for 3 menstrual cycles. The clinical pregnancy rate and embryo implantation rate of the two groups were observed after transplantation; the endometrial thickness and type, resistance index (RI) and pulsatility index (PI) of endometrial blood flow were measured before treatment and one day before transplantation, and adverse reactions was recorded. The clinical pregnancy rate was 37.8% (14/37) in the observation group, which was higher than 16.2% (6/37) in the control group (P<0.05). There was no significant difference in embryo implantation rate between the two groups (P>0.05). One day before transplantation, the endometrial thickness and the proportion of type A in endometrial classification in the two groups were increased compared with those before treatment (P<0.01), and those in the observation group were higher than the control group (P<0.01, P<0.05). The PI and RI of endometrial blood flow in the two groups were lower than those before treatment (P<0.01), and those in the observation group were lower than the control group (P<0.01, P<0.05). During the treatment, 6 patients in the control group had discomfort such as breast distending pain, stomach pain, dizziness and nausea, and there were no adverse reaction in the observation group. On the basis of conventional medication, Tongyuan acupuncture could increase the endometrial thickness, improve endometrial receptivity, improve pregnancy outcome and reduce adverse reactions in patients with RIF of thin endometrial type.


Development and Comparison of Complementary Methods to Study Potential Skin and Inhalational Exposure to Pathogens During Personal Protective Equipment Doffing.
Fluorescent tracers are often used with ultraviolet lights to visibly identify healthcare worker self-contamination after doffing of personal protective equipment (PPE). This method has drawbacks, as it cannot detect pathogen-sized contaminants nor airborne contamination in subjects' breathing zones. A contamination detection/quantification method was developed using 2-µm polystyrene latex spheres (PSLs) to investigate skin contamination (via swabbing) and potential inhalational exposure (via breathing zone air sampler). Porcine skin coupons were used to estimate the PSL swabbing recovery efficiency and limit of detection (LOD). A pilot study with 5 participants compared skin contamination levels detected via the PSL vs fluorescent tracer methods, while the air sampler quantified potential inhalational exposure to PSLs during doffing. Average PSL skin swab recovery efficiency was 40% ± 29% (LOD = 1 PSL/4 cm2 of skin). In the pilot study, all subjects had PSL and fluorescent tracer skin contamination. Two subjects had simultaneously located contamination of both types on a wrist and hand. However, for all other subjects, the PSL method enabled detection of skin contamination that was not detectable by the fluorescent tracer method. Hands/wrists were more commonly contaminated than areas of the head/face (57% vs 23% of swabs with PSL detection, respectively). One subject had PSLs detected by the breathing zone air sampler. This study provides a well-characterized method that can be used to quantitate levels of skin and inhalational contact with simulant pathogen particles. The PSL method serves as a complement to the fluorescent tracer method to study PPE doffing self-contamination.


Theoretical and experimental bases of intraperitoneal chemotherapy.
Pharmacokinetic theory predicts that a large and potentially exploitable concentration difference occurs between the peritoneal cavity and the plasma after many anticancer drugs are administered intraperitoneally in large volume. Unresolved issues remain, particularly concerning the depth of penetration of drugs into tumor nodules growing on peritoneal surfaces. Recent studies in the rat showed a steep concentration gradient of small marker molecules in a variety of normal tissues. The concentration in the stomach and small and large intestines decreased to 10% of the value at the serosal surface in about 0.5 mm or less. Human serum albumin showed deeper penetration, particularly in the diaphragm and anterior abdominal wall.


The Efficacy of Percutaneous Transluminal Angioplasty on the Limb Salvage and Recovery of Symptoms in Patients With Buerger's Disease With Critical Limb Ischemia.
Buerger's disease, or thromboangiitis obliterans, is associated with limb-threatening chronic arterial lesions. In this study, we sought to investigate the efficacy of the percutaneous transluminal angioplasty method for the treatment of critical limb ischemia (CLI) in patients with Buerger's disease in our modest cohort. Patients diagnosed with CLI secondary to Buerger's disease who underwent percutaneous transluminal angioplasty between May 2014 and June 2017 were retrospectively investigated. Patient demographics, presentations, procedural details, responses to percutaneous treatment, complications, limb salvage, wound healing, reinterventions, and early follow-up data were recorded. The cohort included 24 patients with Buerger's disease presenting with CLI observed in 46 limbs. Presentations were gangrene in 12 patients, ulcer formation in 7 patients, and rest pain in the remaining 5 patients. All patients received percutaneous balloon angioplasty, with limb salvage in 21 patients (87.5%). Revascularization was achieved in 87.5% of the destination arteries at the primary intervention and overall technical success rate including reinterventions reached 95.8%. Following the procedures, a total of 22 patients had clinical response with at least ≥1 Rutherford category and mean Rutherford category significantly improved from 5.2 ± 0.74 to 1.6 ± 0.7 (P<.001). Limb salvage rate was 87.5%. Complete wound healing was achieved in all patients with ischemic ulcers at 3.9 ± 2.6 months (range 1-13 months) post revascularization. Mean follow-up duration was 16.07 ± 3.4 months and 6 patients (who were especially subjected to cigarette smoke) required reinterventions. Percutaneous treatment of arterial occlusions in patients with Buerger's disease seems feasible in the current era of improving devices and angioplasty materials. Procedures may be safely performed with good technical and clinical success rates, and without mortality or complications as experience increases.


Effect of posteromedial vertical capsulotomy with medial collateral ligament liberation on intraoperative medial component gap mismatch between extension and mid-flexion during total knee arthroplasty.
The aim of this study was to quantify the effect of posteromedial vertical capsulotomy on intraoperative component gaps and angles from extension through mid-flexion to flexion during total knee arthroplasty (TKA). In the present study, 47 cases of primary posterior-stabilized TKA using the measured resection technique for varus knee osteoarthritis (hip-knee-ankle angles < 0°) were reviewed. Component gaps and angles at 0°, 10°, 45°, 90°, and maximum flexion were measured intraoperatively, before and after posteromedial vertical capsulotomy. Differences in pre- and post-posteromedial vertical capsulotomy medial and lateral component gaps and angles and medial component gap mismatches among knee flexion angles were assessed using the Wilcoxon signed rank test for two paired samples. The medial component gaps at 0° and 10° of flexion of post-posteromedial vertical capsulotomy were significantly greater, exceeding the minimal detectable change, than those pre posteromedial vertical capsulotomy (change of the gap after the procedure at 0° of flexion was 0.7 ± 0.7 mm and at 10° of flexion was 0.8 ± 0.8 mm; all P values < 0.05). The medial component gap mismatches between both 0° and 10°, and 45°, 90°, and maximum flexion were significantly smaller post posteromedial vertical capsulotomy than pre posteromedial vertical capsulotomy, with the values of the change exceeding the minimal detectable change (change of the gap mismatch after the procedure: knee flexion at 0° and 45° was - 0.6 ± 0.9 [mm], at 0° and 90° was 0.7 ± 1.0, at 0° and maximum flexion was - 0.6 ± 1.2, at 10° and 45° was - 0.7 ± 0.9, at 10° and 90° was - 0.8 ± 0.9, at 10° and maximum flexion was - 0.7 ± 1.1; all P values < 0.05). Posteromedial vertical capsulotomy increased the medial component gaps during knee extension but not during mid-flexion or full flexion during posterior-stabilized TKA. Posteromedial vertical capsulotomy improved mild medial component gap mismatch between extension and mid-flexion and full flexion during posterior-stabilized TKA. Surgeons can consider posteromedial vertical capsulotomy when there is intraoperative constriction of the medial component gap during extension in patients undergoing posterior-stabilized TKA.


Focused Ultrasound Central Lateral Thalamotomy for the Treatment of Refractory Neuropathic Pain: Phase I Trial.
Magnetic resonance-guided focused ultrasound (MRgFUS) central lateral thalamotomy (CLT) has not yet been validated for treating refractory neuropathic pain (NP). Our aim was to assess the safety and potential efficacy of MRgFUS CLT for refractory NP. In this prospective, nonrandomized, single-arm, investigator-initiated phase I trial, patients with NP for more than 6 months related to phantom limb pain, spinal cord injury, or radiculopathy/radicular injury and who had undergone at least one previous failed intervention were eligible. The main outcomes were safety profile and pain as assessed using the brief pain inventory, the pain disability index, and the numeric rating scale. Medication use and the functional connectivity of the default mode network (DMN) were also assessed. Ten patients were enrolled, with nine achieving successful ablation. There were no serious adverse events and 12 mild/moderate severity events. The mean age was 50.9 years (SD: 12.7), and the mean symptom duration was 12.3 years (SD: 9.7). Among eight patients with a 1-year follow-up, the brief pain inventory decreased from 7.6 (SD: 1.1) to 3.8 (SD: 2.8), with a mean percent decrease of 46.3 (SD: 40.6) (paired t-test, P = .017). The mean pain disability index decreased from 43.0 (SD: 7.5) to 25.8 (SD: 16.8), with a mean percent decrease of 39.3 (SD: 41.6) (P = .034). Numeric rating scale scores decreased from a mean of 7.2 (SD: 1.8) to 4.0 (SD: 2.8), with a mean percent decrease of 42.8 (SD: 37.8) (P = .024). Patients with predominantly intermittent pain or with allodynia responded better than patients with continuous pain or without allodynia, respectively. Some patients decreased medication use. Resting-state functional connectivity changes were noted, from disruption of the DMN at baseline to reactivation of connectivity between DMN nodes at 3 months. MRgFUS CLT is feasible and safe for refractory NP and has potential utility in reducing symptoms as measured by validated pain scales.