An effective prognostic model in colon adenocarcinoma composed of cuproptosis-related epigenetic regulators.
Background: Colorectal adenocarcinoma (COAD) is a common malignant tumor with little effective prognostic markers. Cuproptosis is a newly discovered mode of cell death that may be related to epigenetic regulators. This study aimed to explore the association between epigenetic regulators and cuproptosis, and to establish a prognostic prediction model for COAD based on epigenetic regulators associated with cuproptosis (EACs). Methods: RNA sequencing data and clinical data of 524 COAD patients were obtained from the TCGA-COAD database, cuproptosis-related genes were from the FerrDb database, and epigenetic-related genes were from databases such as GO and EpiFactors. LASSO regression analysis and other methods were used to screen out epigenetic regulators associated with cuproptosis and prognosis. The risk score of each patient was calculated and the patients were divided into high-risk group and low-risk group. Next, the survival difference, functional enrichment analyses, tumor mutation burden, chemotherapy drug sensitivity and other indicators between the two groups were compared and analyzed. Results: We found 716 epigenetic regulators closely related to cuproptosis, among which 35 genes were related to prognosis of COAD. We further screened out 7 EACs from the 35 EACs to construct a prognostic prediction model. We calculated the risk score of each patient based on these 7 genes, and divided the patients into high-risk group and low-risk group. We found that the overall survival rate and progression-free survival rate of the high-risk group were significantly lower than those of the low-risk group. This model showed good predictive ability in the training set, test set and overall data set. We also constructed a prognostic prediction model based on risk score and other clinical features, and drew the corresponding Nomogram. In addition, we found significant differences between the high-risk group and the low-risk group in tumor mutation burden, chemotherapy drug sensitivity and other clinical aspects. Conclusion: We established an effective predictive prediction model for COAD based on EACs, revealing the association between epigenetic regulators and cuproptosis in COAD. We hope that this model can not only facilitate the treatment decision of COAD patients, but also promote the research progress in the field of cuproptosis.


Evaluation of atrial fibrosis in atrial fibrillation patients with three different methods.
The presence of atrial fibrosis has already been known as a risk factor for atrial fibrillation (AF) development. We aimed to evaluate atrial fibrosis with previously defined three different methods, which were cardiac magnetic resonance imaging (C-MRI), echocardiographic strain imaging, and biomarkers and show the relationship between these methods in patients with AF scheduled for cryoballoon ablation. A total of 30 patients were enrolled. Atrial T1 relaxation durations were measured using C-MRI before the procedure of atrial fibrillation catheter ablation. Fibroblast growth factor-21 (FGF-21) and fibroblast growth factor-23 (FGF-23) levels were measured at serum derived from the femoral artery (Peripheral FGF 21 and 23) and left atrium blood samples (Central FGF 21 and 23) before catheter ablation. Preprocedural transthoracic echocardiography was performed. The median follow-up duration for atrial tachyarrhythmia (ATa) recurrence was 13 (12-18 months) months. The mean ages of the study group were 55.23 ± 12.37 years, and there were 17 (56.7%) female patients in study population. There were negative correlations between post contrast T1 relaxation durations of both posterior and posterosuperior atrium, and central FGF-23 (r: - 0.561; p = 0.003; r:-0.624; p = 0.001; Posterior T1 vs. central FGF-23 levels and Posterosuperior T1 vs central FGF-23 levels, respectively). The positive correlations were observed between postcontrast posterior T1 relaxation durations and left ventricle ejection fraction (r:0.671; p = 0.001); left atrial emptying fraction (r:0.482; p = 0.013); peak atrial longitudinal strain (r:0.605; p = 0.001), and peak atrial contraction strain (r:0.604; p = 0.001). Also negative correlation was observed between postcontrast posterior T1 relaxation durations, and left atrial volume index (r: - 0.467; p = 0.016).


The science of pinning the neck of the femur.
Experiments show that the cancellum of some femoral heads and of the trochanteric region is too weak to support a nail. A simple classification of traumatic femoral neck fractures is presented. These are either vertical or, less commonly, transversely disposed. Either type may be undisplaced, impacted, or displaced. For adequate fixation of undisplaced vertical and all displaced fractures a nail plate is essential. For undisplaced transverse fractures and impacted vertical fractures two screws usually suffice. Impacted transverse fractures require no active treatment. Hammering nails into the femoral head can cause fractures of the articular cartilage, split the femoral head in two, lead to avulsion of articular cartilage under the weight-bearing area, and osteoarthrosis. Early weight-bearing on weak implants can be disastrous. For the unsolved fracture a sliding pin compressing the fragments should be used. In a series of 54 displaced fractures so treated, 50 united. Prosthetic replacement should be reserved for patients with poor-quality bone. Children are best treated with a Coventry lag screw. Non-union after internal fixation in young patients should be salvaged by an excisional osteotomy, but a prosthesis is the answer for elderly patients. Late segmental collapse is explained in terms of contact stresses.


Coupled effects of light and nitrogen source on the urea cycle and nitrogen metabolism over a diel cycle in the marine diatom Thalassiosira pseudonana.
Diatoms are photoautotrophic organisms capable of growing on a variety of inorganic and organic nitrogen sources. Discovery of a complete urea cycle in diatoms was surprising, as this pathway commonly functions in heterotrophic organisms to rid cells of waste nitrogen. To determine how the urea cycle is integrated into cellular nitrogen metabolism and energy management, the centric diatom Thalassiosira pseudonana was maintained in semi-continuous batch cultures on nitrate, ammonium, or urea as the sole nitrogen source, under a 16: 8 light: dark cycle and at light intensities that were low, saturating, or high for growth. Steady-state transcript levels were determined for genes encoding enzymes linked to the urea cycle, urea hydrolysis, glutamine synthesis, pyrimidine synthesis, photorespiration, and energy storage. Transcript abundances were significantly affected by nitrogen source, light intensity and a diel cycle. The impact of N source on differential transcript accumulation was most apparent under the highest light intensity. Models of cellular metabolism under high light were developed based on changes in transcript abundance and predicted enzyme localizations. We hypothesize that the urea cycle is integrated into nitrogen metabolism through its connection to glutamine and in the eventual production of urea. These findings have important implications for nitrogen flow in the cell over diel cycles at surface ocean irradiances.


The Lyn kinase activator MLR-1023 is a novel insulin receptor potentiator that elicits a rapid-onset and durable improvement in glucose homeostasis in animal models of type 2 diabetes.
MLR-1023 [Tolimidone; CP-26154; 2(1H)-pyrimidinone, 5-(3-methylphenoxy)] is an allosteric Lyn kinase activator that reduces blood glucose levels in mice subjected to an oral glucose tolerance test (J Pharmacol Exp Ther 342:15-22, 2012). The current studies were designed to define the role of insulin in MLR-1023-mediated blood glucose lowering, to evaluate it in animal models of type 2 diabetes, and to compare it to the activities of selected existing diabetes therapeutics. Results from these studies show that in an acute oral glucose tolerance test MLR-1023 evoked a dose-dependent blood glucose-lowering response that was equivalent in magnitude to that of metformin without eliciting a hypoglycemic response. In streptozotocin-treated, insulin-depleted mice, MLR-1023 administration did not affect blood glucose levels. However, MLR-1023 potentiated the glucose-lowering activity of exogenously administered insulin, showing that MLR-1023-mediated blood glucose lowering was insulin-dependent. In a hyperinsulinemic/euglycemic clamp study, orally administered MLR-1023 increased the glucose infusion rate required to sustain blood glucose levels, demonstrating that MLR-1023 increased insulin receptor sensitivity. In chronically treated db/db mice, MLR-1023 elicited a dose-dependent and durable glucose-lowering effect, reduction in HbA1c levels and preservation of pancreatic β-cells. The magnitude of effect was equivalent to that seen with rosiglitazone but with a faster onset of action and without causing weight gain. These studies show that MLR-1023 is an insulin receptor-potentiating agent that produces a rapid-onset and durable blood glucose-lowering activity in diabetic animals.


Systematic Comparison of Heatmapping Techniques in Deep Learning in the Context of Diabetic Retinopathy Lesion Detection.
Heatmapping techniques can support explainability of deep learning (DL) predictions in medical image analysis. However, individual techniques have been mainly applied in a descriptive way without an objective and systematic evaluation. We investigated comparative performances using diabetic retinopathy lesion detection as a benchmark task. The Indian Diabetic Retinopathy Image Dataset (IDRiD) publicly available database contains fundus images of diabetes patients with pixel level annotations of diabetic retinopathy (DR) lesions, the ground truth for this study. Three in advance trained DL models (ResNet50, VGG16 or InceptionV3) were used for DR detection in these images. Next, explainability was visualized with each of the 10 most used heatmapping techniques. The quantitative correspondence between the output of a heatmap and the ground truth was evaluated with the Explainability Consistency Score (ECS), a metric between 0 and 1, developed for this comparative task. In case of the overall DR lesions detection, the ECS ranged from 0.21 to 0.51 for all model/heatmapping combinations. The highest score was for VGG16+Grad-CAM (ECS = 0.51; 95% confidence interval [CI]: [0.46; 0.55]). For individual lesions, VGG16+Grad-CAM performed best on hemorrhages and hard exudates. ResNet50+SmoothGrad performed best for soft exudates and ResNet50+Guided Backpropagation performed best for microaneurysms. Our empirical evaluation on the IDRiD database demonstrated that the combination DL model/heatmapping affects explainability when considering common DR lesions. Our approach found considerable disagreement between regions highlighted by heatmaps and expert annotations. We warrant a more systematic investigation and analysis of heatmaps for reliable explanation of image-based predictions of deep learning models.


Association between mitral and aortic valve calcification and preferential left or right coronary artery disease.
Mitral annular calcification (MAC) and aortic valve calcification (AVC) are predictive of coronary artery disease (CAD). However, no data exist concerning the association between preferential CAD side localization to the left or right coronary arteries and MAC or AVC. A cohort analysis was performed of 1,000 consecutive coronary angiographies recorded in patients with CAD. The angiographies were divided according to the distribution of CAD to the isolated right coronary tree disease, left coronary tree disease, or both. The echocardiograms were reviewed for MAC, AVC or combined valvular calcification (CVC). Significant CAD (lumenal stenosis > 70%) was observed in 688 patients, among whom 167 had isolated (right or left) CAD and 521 double-sided coronary tree disease. Valvular calcification (VC) was observed in 70 (42%) of the isolated CAD patients; of these, 41 had isolated left CAD and 29 isolated right CAD. Among the isolated left CAD patients with VC, 13 (32%) had AVC, 22 (53%) had CVC, and only six (15%) had MAC (p < 0.01). Among the isolated right CAD patients with VC, 18 (62%) had MAC, nine (31%) had CVC, and only two (7%) had AVC (p < 0.01). VC was observed in 266 patients (51%) with mixed CAD; of these, 152 (57%) had CVC, 103 (39%) had AVC, and 11 (4%) had MAC (p < 0.01). Isolated left CAD is associated with AVC or CVC more frequently than with MAC. In contrast, isolated right CAD is associated with MAC or CVC, but rarely with AVC.


Electron-phonon scattering limited hole mobility at room temperature in a MoS2 monolayer: first-principles calculations.
Based on first-principles calculations and iterative solution of the Boltzmann transport equation, we theoretically study the room temperature mobility (RTM) of a valence band hole in a MoS2 monolayer (ML) limited by electron-phonon (e-ph) scattering. The hole mobility obtained by us is 26.0 cm2 V-1 s-1 at 300 K. This is a value much closer to the experimental result (about 40.0 cm2 V-1 s-1). In contrast, the semi-empirical estimate based on the deformational potential (DP) model in previous literature gave a value of 200.5 cm2 V-1 s-1, far away from the experimental data. By a detailed analysis, we find that unlike the case of conduction band electrons, the intervalley scattering realized by longitudinal acoustic (LA) phonons plays a dominant role in influencing the hole mobility. And this is the main reason for the DP model failing to give a quantitative estimate of the hole RTM in MoS2-ML.


Atomic Layer Deposited Lithium Silicates as Solid-State Electrolytes for All-Solid-State Batteries.
Development of solid-state electrolyte (SSE) thin films is a key toward the fabrication of all-solid-state batteries (ASSBs). However, it is challenging for conventional deposition techniques to deposit uniform and conformal SSE thin films in a well-controlled fashion. In this study, atomic layer deposition (ALD) was used to fabricate lithium silicate thin films as a potential SSE for ASSBs. Lithium silicates thin films were deposited by combining ALD Li2O and SiO2 subcycles using lithium tert-butoxide, tetraethylorthosilane, and H2O as precursors. Uniform and self-limiting growth was achieved at temperatures between 225 and 300 °C. X-ray absorption spectroscopy analysis disclosed that the as-deposited lithium silicates were composed of SiO4 tetrahedron structure and lithium oxide as the network modifier. X-ray photoelectron spectroscopy confirmed the chemical states of Li in the thin films were the same with that in standard lithium silicate. With one to one subcycle of Li2O and SiO2 the thin films had a composition close to Li4SiO4 whereas one more subcycle of Li2O delivered a higher lithium content. The lithium silicate thin film prepared at 250 °C exhibited an ionic conductivity of 1.45× 10-6 S cm-1 at 373 K. The high ionic conductivity of lithium silicate was due to the higher lithium concentration and lower activation energy.


Extracorporeal Shock Waves Increase Markers of Cellular Proliferation in Bronchial Epithelium and in Primary Bronchial Fibroblasts of COPD Patients.
Chronic obstructive pulmonary disease (COPD) is due to structural changes and narrowing of small airways and parenchymal destruction (loss of the alveolar attachment as a result of pulmonary emphysema), which all lead to airflow limitation. Extracorporeal shock waves (ESW) increase cell proliferation and differentiation of connective tissue fibroblasts. To date no studies are available on ESW treatment of human bronchial fibroblasts and epithelial cells from COPD and control subjects. We obtained primary bronchial fibroblasts from bronchial biopsies of 3 patients with mild/moderate COPD and 3 control smokers with normal lung function. 16HBE cells were also studied. Cells were treated with a piezoelectric shock wave generator at low energy (0.3 mJ/mm2, 500 pulses). After treatment, viability was evaluated and cells were recultured and followed up for 4, 24, 48, and 72 h. Cell growth (WST-1 test) was assessed, and proliferation markers were analyzed by qRT-PCR in cell lysates and by ELISA tests in cell supernatants and cell lysates. After ESW treatment, we observed a significant increase of cell proliferation in all cell types. C-Kit (CD117) mRNA was significantly increased in 16HBE cells at 4 h. Protein levels were significantly increased for c-Kit (CD117) at 4 h in 16HBE (p < 0.0001) and at 24 h in COPD-fibroblasts (p = 0.037); for PCNA at 4 h in 16HBE (p = 0.046); for Thy1 (CD90) at 24 and 72 h in CS-fibroblasts (p = 0.031 and p = 0.041); for TGFβ1 at 72 h in CS-fibroblasts (p = 0.038); for procollagen-1 at 4 h in COPD-fibroblasts (p = 0.020); and for NF-κB-p65 at 4 and 24 h in 16HBE (p = 0.015 and p = 0.0002). In the peripheral lung tissue of a representative COPD patient, alveolar type II epithelial cells (TTF-1+) coexpressing c-Kit (CD117) and PCNA were occasionally observed. These data show an increase of cell proliferation induced by a low dosage of extracorporeal shock waves in 16HBE cells and primary bronchial fibroblasts of COPD and control smoking subjects.


Pulmonary fibrosis with predominant CD8 lymphocytic alveolitis and anti-Jo-1 antibodies.
Interstitial lung disease (ILD) is a complication of polymyositis (PM) and dermatomyositis (DM). It often manifests itself in association with myositis-specific antisynthetase autoantibodies, among which anti-Jo-1 antibodies are the most commonly encountered. In contrast, ILD associated with anti-Jo-1 antibodies without muscle involvement is rare and not well characterized. We report four patients presenting with ILD associated with anti-Jo-1 antibodies. Histological findings of transbronchial biopsies disclosed a pattern consistent with nonspecific interstitial pneumonitis, a CD8+ lymphocytosis was found in bronchoalveolar lavage. Only one of these patients developed an "antisynthetase syndrome" with PM, after nearly 2 yrs of severe ILD. The clinical conditions of all four cases showed stabilization or improvement when cyclosporine was added to their immunosuppressive treatment. These cases confirm that a CD8+ lymphocytic interstitial lung disease may be the first, and sole manifestation of autoimmune disease associated with anti-Jo-1 antibodies. Furthermore, they suggest that this form of interstitial lung disease apparently has a poor response to steroids and cytotoxic drugs, but may respond to moderate doses of cyclosporine and azathioprine in addition to low doses of steroids.


Six-membered ring chelate complexes of Ru(II): structural and photophysical effects.
The structural and photophysical properties of Ru(II)-polypyridyl complexes with five- and six-membered chelate rings were studied for two bis-tridentate and two tris-bidentate complexes. The photophysical effect of introducing a six-membered chelate ring is most pronounced for the tridentate complex, leading to a room-temperature excited-state lifetime of 810 ns, a substantial increase from 180 ns for the five-membered chelate ring model complex. Contrasting this, the effect is the opposite in tris-bidentate complexes, in which the lifetime decreases from 430 ns to around 1 ns in going from a five-membered to six-membered chelate ring. All of the complexes were studied spectroscopically at both 80 K and ambient temperatures, and the temperature dependence of the excited-state lifetime was investigated for both of the bis-tridentate complexes. The main reason for the long excited-state lifetime in the six-membered chelate ring bis-tridentate complex was found to be a strong retardation of the activated decay via metal-centered states, largely due to an increased ligand field splitting due to the complex having a more-octahedral geometry.


Behavior therapy and the transdermal nicotine patch: effects on cessation outcome, affect, and coping.
The process and outcome of a smoking cessation program using behavior therapy alone (BT) or behavior therapy plus the nicotine patch (BTP) was studied in 64 participants. Participants quit smoking on a target date after a period of ad libitum smoking, cognitive-behavior therapy preparing them for cessation, and behavioral rehearsal for high-risk situations, including stress management, and coping with negative affect. Abstinence was significantly higher for the BTP group versus the BT group from the end of behavioral treatment (79% vs. 63%) through the 3-month follow-up (p < .01), with the effects weakening at the 6- (p = .06) and 12-month marks (p = 38% vs. 22%). More general distress was observed among BT versus BTP participants (i.e., increased withdrawal, tension, fatigue, and coping frequency with decreased coping effort; coping-to-urge ratio). The coping behavior of the BTP group may have been more effective than that of the BT group, as indicated by their significantly higher level of self-efficacy.


Preparation of surface molecularly imprinted Ru-complex catalysts for asymmetric transfer hydrogenation in water media.
Molecularly imprinted Ru-complex catalysts acting in water were prepared on a SiO(2) surface by molecular imprinting of a SiO(2)-supported Ru-complex using organic polymers as surface matrix overlayers. (R)-1-(o-fluorophenyl)ethanol, which is one of the hydrogenated products of o-fluoroacetophenone, was imprinted on the supported Ru-complex as a template, and an active Ru-complex with a shape-selective reaction space (molecularly imprinted cavity) was prepared inside the wall of the hydrophobic organic polymer matrix overlayers. Structures of the SiO(2)-supported and molecularly imprinted Ru catalysts were characterized by means of solid-state NMR, XPS, XRF, ICP, UV/vis, XAFS, TGA, and SEM. The molecularly imprinted Ru catalysts exhibited fine shape selectivity and enantioselectivity for the asymmetric transfer hydrogenation of o-fluoroacetophenone and its derivatives.


Factors affecting willingness to share electronic health data among California consumers.
Robust technology infrastructure is needed to enable learning health care systems to improve quality, access, and cost. Such infrastructure relies on the trust and confidence of individuals to share their health data for healthcare and research. Few studies have addressed consumers' views on electronic data sharing and fewer still have explored the dual purposes of healthcare and research together. The objective of the study is to explore factors that affect consumers' willingness to share electronic health information for healthcare and research. This study involved a random-digit dial telephone survey of 800 adult Californians conducted in English and Spanish. Logistic regression was performed using backward selection to test for significant (p-value ≤ 0.05) associations of each explanatory variable with the outcome variable. The odds of consent for electronic data sharing for healthcare decreased as Likert scale ratings for EHR impact on privacy worsened, odds ratio (OR) = 0.74, 95% CI [0.60, 0.90]; security, OR = 0.80, 95% CI [0.66, 0.98]; and quality, OR = 0.59, 95% CI [0.46-0.75]. The odds of consent for sharing for research was greater for those who think EHR will improve research quality, OR = 11.26, 95% CI [4.13, 30.73]; those who value research benefit over privacy OR = 2.72, 95% CI [1.55, 4.78]; and those who value control over research benefit OR = 0.49, 95% CI [0.26, 0.94]. Consumers' choices about electronically sharing health information are affected by their attitudes toward EHRs as well as beliefs about research benefit and individual control. Design of person-centered interventions utilizing electronically collected health information, and policies regarding data sharing should address these values of importance to people. Understanding of these perspectives is critical for leveraging health data to support learning health care systems.


Clinicohistoradiological and surgical outcome in diffuse midline glioma.
Diffuse Midline Glioma (DMG) with H3K27M mutation is a rare and aggressive midline high grade glioma with a predominant astrocytic differentiation and K27M mutation in either H3F3A or HIST1H3B/C. This tumor is more common in children than in adults. The current study was aimed to determine clinicohistoradiological and surgical outcome of patients who have undergone surgery for DMG and study disease severity of patients with DMG. This is an observational study in which 29 DMG patients were evaluated for clinicohistoradiological and surgical outcomes by assessing the pre and postoperative neurological status. Survival duration was significantly high in patients with age > 18 years (p = 0.02). Patients who had undergone Radiation Therapy showed higher survival rate (p = 0.05) and the cases with low levels of Ki 67 index had improved post operative outcome (p = 0.002). DMG with H3K27M mutation in newly classified Central Nervous System tumor are WHO grade IV Tumors, comprising H3K27M mutation as molecular marker for diagnosis and related with a poor prognosis.


Cough . 2: Chronic cough in children.
Chronic cough is a common problem in childhood. Viral infections are the most prevalent cause, but other rarer disorders should be excluded whenever cough appears unusually severe and/or frequent, and when there is evidence of failure to thrive and growth retardation. The younger the child, the more the need to exclude underlying disease at an early stage. Passive smoking is an important contributor to chronic cough in children. Chronic productive cough with purulent sputum is always reason for concern in children and is not common as a symptom of asthma. More or less specific diagnoses in children include cystic fibrosis, aspirated foreign body, congenital anatomic abnormalities and primary ciliary dyskinesia.


Ideomotor apraxia: behavioral dimensions and neuroanatomical basis.
Ideomotor apraxia, disordered movement execution to command, commonly follows left-hemisphere damage, implying left-hemisphere dominance for certain kinds of movements. To delineate this dominance we used different command modalities to elicit meaningful movements and tested imitation of nonsense movements. Twenty-seven patients with unilateral hemispheric stroke and 10 age-matched controls were evaluated. Patients with left-hemisphere damage performed both meaningful and nonsense movements poorer than the other study groups; thus, the meaningfulness of the movements is irrelevant for the left-hemisphere motor dominance. The performance varied, however, with the command modality and movement type. Based on this and earlier studies we posit that the left-hemisphere motor dominance is determined by the artificiality of the test situation (it concerns movements performed to command and out of the natural context) and increased spatial and temporal complexity of the demanded movements. No association between the lesion locus within the left hemisphere and the severity of the ideomotor apraxia was found.


Pathophysiology of cholangiopathies.
The diseases of the intrahepatic biliary tree are a large group of potentially evolutive congenital and acquired liver disorders affecting both the adult and pediatric populations. They represent a relevant cause of liver-related morbidity and mortality and an important indication for liver transplantation, particularly in children. While the practical approach to patients affected by biliary tree diseases has not significantly changed yet, the conceptual approach to the pathophysiology of cholangiopathies has witnessed important advances that will be discussed. The primary cell target of the pathogenetic sequence of these disorders is the biliary epithelium. Cholangiocytes have multifaceted functions, not limited to bile production. Their capability to secrete a range of different pro-inflammatory mediators, cytokines, and chemokines indicates a major role of cholangiocytes in the inflammatory reaction. Furthermore, paracrine secretion of growth factors and peptides mediates an extensive cross-talk with other liver cell types, including hepatocytes, stellate, and endothelial and inflammatory cells. Cholangiopathies share a number of pathogenetic mechanisms, including inflammation, cholestasis, fibrosis, apoptosis, altered development, and neoplastic transformation. These basic disease mechanisms will be discussed in detail, along with the distinct features of a number of cholangiopathies. Furthermore, an increase in the biliary cell compartment is a common response to many forms of liver injury, from cholangiopathies to viral and fulminant hepatitis. Elucidation of these pathophysiologic mechanisms will likely provide clues for future therapeutic strategies. Furthermore, understanding the role of cholangiocytes in liver regeneration/repair and the mechanisms of cholangiocyte activation and their relationship with liver progenitor cell will be of further interest.


Serum dioxin concentrations and time to pregnancy.
Pollution may play a role in population trends of declining semen quality and regional differences in time to pregnancy (TTP) in industrialized societies. Dioxins including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) have been suspected. In 1976, an explosion near Seveso, Italy resulted in the highest TCDD exposure known in residential populations. Twenty years later, we conducted a retrospective cohort study, the Seveso Women's Health Study. Of 981 participants, 472 women attempted pregnancy after the explosion, and 278 delivered a livebirth not associated with contraceptive failure. Individual serum TCDD levels were measured from samples collected soon after the explosion and extrapolated to the conception attempt. We examined the relation of TCDD levels to time to pregnancy (parameterized as the monthly probability of conception within the first 12 months of trying) and to infertility (defined as conception after at least 12 months of trying). We modeled fecundability with discrete-time Cox proportional hazards regression, and we modeled fertility with logistic regression. We tested the sensitivity of the conclusions to differing definitions of eligibility and outcome. Median TCDD level was 50 parts per trillion, median time to pregnancy was 2 months, and 17% reported taking 12 or more months to conceive. For every 10-fold increase in serum TCDD, we observed a 25% increase in time to pregnancy (adjusted-fecundability odds ratio = 0.75 [95% confidence interval (CI) = 0.60-0.95]) and about a doubling in odds of infertility (adjusted odds ratio = 1.9 [95% CI = 1.1-3.2]). Results were similar for extrapolated TCDD and sensitivity analyses. We found dose-related increases in TTP and infertility associated with individual serum TCDD levels in the women from Seveso, Italy. These findings may have implications for fertility in industrialized areas.


Impact of Aprotinin - A Proteolytic Enzyme on Postsurgical Symptoms in Patients Undergoing Third Molar Surgeries.
Dealing with postoperative pain and inflammation remains an arena for never ending research. Different agents have been the subject of many studies to prevent the occurrence of unpleasant postoperative sequel. Extraction of third molars is often associated with significant deterioration in oral health related quality of life (physical, social and psychological) in immediate postoperative period. The complaints of pain, swelling and limitation of mouth opening, which ensue as a result of acute inflammatory response, are frequent consequences of postsurgical procedures involving extraction of impacted 3(rd) molars. Aprotinin, a naturally occurring protease inhibitor was assessed for its effectiveness in plummeting postsurgical pain and swelling, after surgical removal of impacted 3(rd) molars. Thirty randomly selected adult patients age ranging from 16-35 years, who required simultaneous surgical removal of bilateral impacted mandibular third molars were recruited. Before the surgical procedure, randomly selected side of the patient was injected with 1 m of 10,000 Kallikrein Inactivator Units (KIU) of aprotinin sub-mucosally around the surgical site and the contra lateral side with 1ml of isotonic saline as a control following which adequate local anaesthesia was obtained. The surgical removal of impacted 3(rd) molars was conducted in a similar manner on both test and control sides on all patients. Postoperatively, the patients were evaluated for pain and swelling for one week i.e., 1(st), 2(nd) and 7(th) day. It was observed that there was marked clinical reduction in postoperative pain and swelling. There were no adverse affects observed after using aprotinin. Since, the current pharmacologic agents being used have adverse effects and associated morbidity which still pose a problem, aprotinin a naturally occurring agent could be efficiently used after surgical extraction of 3(rd) molars in management of postsurgical symptoms and improve patient comfort and quality of life. In future, further studies with use of aprotinin in a large number of patients and comparative studies with other drugs are required.


[Reactivity and immunogenicity of the Sonne dysentery detergent monovaccine].
The authors elaborated a method of isolation under industrial conditions of Shigella sonnei surface antigens by the treatment of the culture with a detergent (sodium laurylsulfate). The antigens obtained possessed a much lesser toxicity in comparison with the tryptic ones. A method of preparation of a detergent Sonne monovaccine with an increased antigen content (1.3 mg/ml) was described. Experimental series of the preparation were tested on 38 volunteers. A lower reactogenicity of the detergent vaccine (in comparison with the tryptic) and its marked immunological efficacy was demonstrated.


Sleep Disturbances Related to Environmental Noise.
The permanently open auditory channel and the ability of the brain to process incoming acoustical stimuli even while asleep and to respond adequately is the essential precondition for noise-induced sleep disturbances which are regarded as the most deleterious effects of noise. In the past, research was mainly focused on the detection and description of the various effects of noise, on the influence of personal and environmental factors, on the determination of dose-response relations and the definition of critical noise loads, above which noise becomes intolerable. These limits are, however, as yet only tentative or applicable for a very few situations and need to be verified or revised. The present paper is focused on the priorities for future research. These are in particular 1) the causal linkage between environmental noise, primary and secondary effects on the one hand and the hypothesized contribution to multifactorial chronic diseases, to chronic annoyance, and to permanent behavioural alterations on the other hand, 2) the identification of the causes for the great discrepancies between the small effects determined in the field and the large responses recorded in the laboratory, 3) temporal aspects such as sleep at unusual times (day sleep after nightshifts), definition of night-time and day-time, 4) the significance of the shoulder hours for subsequent sleep, 5) the individual vulnerability, 6) the accumulation of data from different studies.


Deficiency of complement receptors CR2/CR1 in Cr2⁻/⁻ mice reduces the extent of secondary brain damage after closed head injury.
Complement activation at the C3 convertase level has been associated with acute neuroinflammation and secondary brain injury after severe head trauma. The present study was designed to test the hypothesis that Cr2-/- mice, which lack the receptors CR2/CD21 and CR1/CD35 for complement C3-derived activation fragments, are protected from adverse sequelae of experimental closed head injury. Adult wild-type mice and Cr2-/- mice on a C57BL/6 genetic background were subjected to focal closed head injury using a standardized weight-drop device. Head-injured Cr2-/- mice showed significantly improved neurological outcomes for up to 72 hours after trauma and a significantly decreased post-injury mortality when compared to wild-type mice. In addition, the Cr2-/- genotype was associated with a decreased extent of neuronal cell death at seven days post-injury. Western blot analysis revealed that complement C3 levels were reduced in the injured brain hemispheres of Cr2-/- mice, whereas plasma C3 levels remained unchanged, compared to wild-type mice. Finally, head-injured Cr2-/- had an attenuated extent of post-injury C3 tissue deposition, decreased astrocytosis and microglial activation, and attenuated immunoglobulin M deposition in injured brains compared to wild-type mice. Targeting of these receptors for complement C3 fragments (CR2/CR1) may represent a promising future approach for therapeutic immunomodulation after traumatic brain injury.


Evidence of Air and Surface Contamination with SARS-CoV-2 in a Major Hospital in Portugal.
As the third wave of the COVID-19 pandemic hit Portugal, it forced the country to reintroduce lockdown measures due to hospitals reaching their full capacities. Under these circumstances, environmental contamination by SARS-CoV-2 in different areas of one of Portugal's major Hospitals was assessed between 21 January and 11 February 2021. Air samples (n = 44) were collected from eleven different areas of the Hospital (four COVID-19 and seven non-COVID-19 areas) using Coriolis® μ and Coriolis® Compact cyclone air sampling devices. Surface sampling was also performed (n = 17) on four areas (one COVID-19 and three non-COVID-19 areas). RNA extraction followed by a one-step RT-qPCR adapted for quantitative purposes were performed. Of the 44 air samples, two were positive for SARS-CoV-2 RNA (6575 copies/m3 and 6662.5 copies/m3, respectively). Of the 17 surface samples, three were positive for SARS-CoV-2 RNA (200.6 copies/cm2, 179.2 copies/cm2, and 201.7 copies/cm2, respectively). SARS-CoV-2 environmental contamination was found both in air and on surfaces in both COVID-19 and non-COVID-19 areas. Moreover, our results suggest that longer collection sessions are needed to detect point contaminations. This reinforces the need to remain cautious at all times, not only when in close contact with infected individuals. Hand hygiene and other standard transmission-prevention guidelines should be continuously followed to avoid nosocomial COVID-19.


Diagnostic accuracy of laparoscopy following computed tomography (CT) scanning for assessing the resectability with curative intent in pancreatic and periampullary cancer.
Surgical resection is the only potentially curative treatment for pancreatic and periampullary cancer. A considerable proportion of patients undergo unnecessary laparotomy because of underestimation of the extent of the cancer on computed tomography (CT) scanning. Laparoscopy can detect metastases not visualised on CT scanning, enabling better assessment of the spread of cancer (staging of cancer). There has been no systematic review or meta-analysis assessing the role of diagnostic laparoscopy in assessing the resectability with curative intent in patients with pancreatic and periampullary cancer. To determine the diagnostic accuracy of diagnostic laparoscopy performed as an add-on test to CT scanning in the assessment of curative resectability in pancreatic and periampullary cancer. We searched the Cochrane Register of Diagnostic Test Accuracy Studies, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE via PubMed, EMBASE via OvidSP (from inception to 13 September 2012), and Science Citation Index Expanded (from 1980 to 13 September 2012). We included diagnostic accuracy studies of diagnostic laparoscopy in patients with potentially resectable pancreatic and periampullary cancer on CT scan, where confirmation of liver or peritoneal involvement was by histopathological examination of suspicious (liver or peritoneal) lesions obtained at diagnostic laparoscopy or laparotomy. We accepted any criteria of resectability used in the studies. We included studies irrespective of language, publication status, or study design (prospective or retrospective). We excluded case-control studies. Two authors independently performed data extraction and quality assessment using the QUADAS-2 tool. The specificity of diagnostic laparoscopy in all studies was 1 because there were no false positives since laparoscopy and the reference standard are one and the same if histological examination after diagnostic laparoscopy is positive. Therefore, the sensitivities were meta-analysed using a univariate random-effects logistic regression model. The probability of unresectability in patients who had a negative laparoscopy (post-test probability for patients with a negative test result) was calculated using the median probability of unresectability (pre-test probability) from the included studies and the negative likelihood ratio derived from the model (specificity of 1 assumed). The difference between the pre-test and post-test probabilities gave the overall added value of diagnostic laparoscopy compared to the standard practice of CT scan staging alone. Fifteen studies with a total of 1015 patients were included in the meta-analysis. Only one study including 52 patients had a low risk of bias and low applicability concern in the patient selection domain. The median pre-test probability of unresectable disease after CT scanning across studies was 40.3% (that is 40 out of 100 patients who had resectable cancer after CT scan were found to have unresectable disease on laparotomy). The summary sensitivity of diagnostic laparoscopy was 68.7% (95% CI 54.3% to 80.2%). Assuming a pre-test probability of 40.3%, the post-test probability of unresectable disease for patients with a negative test result was 0.17 (95% CI 0.12 to 0.24). This indicates that if a patient is said to have resectable disease after diagnostic laparoscopy and CT scan, there is a 17% probability that their cancer will be unresectable compared to a 40% probability for those receiving CT alone.A subgroup analysis of patients with pancreatic cancer gave a summary sensitivity of 67.9% (95% CI 41.1% to 86.5%). The post-test probability of unresectable disease after being considered resectable on both CT and diagnostic laparoscopy was 18% compared to 40% for those receiving CT alone. Diagnostic laparoscopy may decrease the rate of unnecessary laparotomy in patients with pancreatic and periampullary cancer found to have resectable disease on CT scan. On average, using diagnostic laparoscopy with biopsy and histopathological confirmation of suspicious lesions prior to laparotomy would avoid 23 unnecessary laparotomies in 100 patients in whom resection of cancer with curative intent is planned.


Individual cognitive-behavioral therapy and behavioral couples therapy in alcohol use disorder: a comparative evaluation in community-based addiction treatment centers.
Alcohol abuse serves as a chronic stressor between partners and has a deleterious effect on relationship functioning. Behavioral Couples Therapy (BCT) for alcohol dependence, studied as an adjunct to individual outpatient counseling, has shown to be effective in decreasing alcohol consumption and enhancing marital functioning, but no study has directly tested the comparative effectiveness of stand-alone BCT versus an individually focused cognitive-behavioral therapy (CBT) in a clinical community sample. The present study is a randomized clinical trial evaluating the effectiveness of stand-alone BCT (n = 30) compared to individual CBT (n = 34) in the treatment of alcohol use disorders in community treatment centers in Dutch male and female alcoholics and their partners. Results show both BCT and CBT to be effective in changing drinking behavior after treatment. BCT was not found to be superior to CBT. Marital satisfaction of the spouse increased significantly in the BCT condition but not in the CBT condition, the differences being significant at the post-test. Patients' self-efficacy to withstand alcohol-related high-risk situations increased significantly in both treatment conditions, but more so in CBT than in BCT after treatment. Treatment involvement of the spouse did not increase retention. Regular practitioners in community treatment centers can effectively deliver both treatments. Stand-alone BCT is as effective as CBT in terms of reduced drinking and to some extent more effective in terms of enhancing relationship satisfaction. However, BCT is a more costly intervention, given that treatment sessions lasted almost twice as long as individual CBT sessions.


Measuring Distress Levels of Refugee Women to Enhance Community-Based Psycho-social Interventions.
The purpose of this investigation was to gain an understanding of the levels of distress in resettled refugee women as a basis for the development of improved community based interventions to enhance the resettlement experience. A convenience sample of female refugees (n = 23) in a southern US city were interviewed using a socio-demographic questionnaire; the Refugee Health Screener-15 (RHS-15); and semi-structured, exploratory questions. Results showed consistently high levels of distress, indicated by responses to the RHS-15. Women ages 19-25 and over 50 displayed the highest risk. Categories of stressors included a lack of access to health and psycho-social support services, difficulty with communicating in the English language, and social isolation. In addition, women reported that a lack of trained interpreters contributes to reduced access to health and social service, causing further distress.


Terminals of reserpine-sensitive vasopressin-neurophysin neurons in the external layer of the rat median eminence.
The indirect immunofluorescence technique was used to examine the pharmacology associated with reserpine-induced alterations in vasopressin and neurophysin (VP/NP) immunoreactivity in the external layer of the median eminence in the rat. Twenty-four hours after injection of reserpine, a selective, marked depletion of VP/NP immunoreactivity from the external layer is apparent. Pretreatment with the monoamine oxidase inhibitors, pargyline and tranylcypromine, prevents the depleting effect of reserpine, indicating that the acute effect of reserpine is mediated by monoamines. Acute intraventricular treatment with 6-hydroxydopamine, but not 5,7-dihydroxytryptamine, mimicked the reserpine effect, suggesting that catecholamines mediate reserpine depletion of VP/NP immunoreactivity from the external layer. The experimental results are consistent with a regulatory model in which catecholamines tonically inhibit VP/NP release from terminals in the external layer of the median eminence. Although the studies do not definitively determine the functional relationship between VP and ACTH, the anatomical location of these terminals, the dramatic change in the VP/NP content of these terminals in response to reserpine, and the lack of a response to dehydration suggest that this pool of vasopressin may contribute to ACTH hypersecretion in response to reserpine.


Concurrent chemoradiation for vaginal cancer.
It is not known whether the addition of chemotherapy to radiation therapy improves outcomes in primary vaginal cancer. Here, we review clinical outcomes in patients with primary vaginal cancer treated with radiation therapy (RT) or concurrent chemoradiation therapy (CRT). Seventy-one patients with primary vaginal cancer treated with definitive RT with or without concurrent chemotherapy at a single institution were identified and their records reviewed. A total of 51 patients were treated with RT alone; 20 patients were treated with CRT. Recurrences were analyzed. Overall survival (OS) and disease-free survival (DFS) rates were estimated using the Kaplan-Meier method. Cox regression analysis was performed. The median age at diagnosis was 61 years (range, 18-92 years) and the median follow-up time among survivors was 3.0 years. Kaplan-Meier estimates for OS and DFS differed significantly between the RT and CRT groups (3-yr OS = 56% vs. 79%, log-rank p = 0.037; 3-yr DFS = 43% vs. 73%, log-rank p = 0.011). Twenty-three patients (45%) in the RT group had a relapse at any site compared to 3 (15%) in the CRT group (p = 0.027). With regard to the sites of first relapse, 10 patients (14%) had local only, 4 (6%) had local and regional, 9 (13%) had regional only, 1 (1%) had regional and distant, and 2 (3%) had distant only relapse. On univariate analysis, the use of concurrent chemotherapy, FIGO stage, tumor size, and date of diagnosis were significant predictors of DFS. On multivariate analysis, the use of concurrent chemotherapy remained a significant predictor of DFS (hazard ratio 0.31 (95% CI, 0.10-0.97; p = 0.04)). Vaginal cancer results in poor outcomes. Adequate radiation dose is essential to ensure curative management. Concurrent chemotherapy should be considered for vaginal cancer patients.


Oligodendroglial progenitor cell therapy limits central neurological deficits in mice with metachromatic leukodystrophy.
This work describes the first successful oligodendrocyte-based cell therapy for presymptomatic arylsulfatase A (ARSA) null neonate mice, a murine model for human metachromatic leukodystrophy (MLD). We found that oligodendrocyte progenitors (OLPs) engrafted and survived into adulthood when transplanted in the neonatal MLD brain. Transplanted cells integrated nondisruptively, did not produce tumors, and survived as proteolipid protein- and MBP-positive postmitotic myelinating oligodendrocytes (OLs) intermingled with endogenous MLD OLs within the adult MLD white matter. Transplanted MLD mice had reduced sulfatide accumulation in the CNS, increased brain ARSA activity, and full prevention of the electrophysiological and motor deficits that characterize untreated MLD mice. Our results provide direct evidence that healthy OLPs can tolerate the neurotoxic accumulation of sulfatides that evolves during the postnatal development of the MLD brain and contribute to OL cell replacement to limit the accumulation of sulfatides and the evolution of CNS defects in this lysosomal storage disease mouse model.


Eight hours of nocturnal 915 MHz radiofrequency identification (RFID) exposure reduces urinary levels of melatonin and its metabolite via pineal arylalkylamine N-acetyltransferase activity in male rats.
We investigated the effects of whole-body exposure to the 915 MHz radiofrequency identification (RFID) on melatonin biosynthesis and the activity of rat pineal arylalkylamine N-acetyltransferase (AANAT). Rats were exposed to RFID (whole-body specific absorption rate, 4 W/kg) for 8 h/day, 5 days/week, for weeks during the nighttime. Total volume of urine excreted during a 24-h period was collected after RFID exposure. Urinary melatonin and 6-hydroxymelatonin sulfate (6-OHMS) was measured by gas chromatography-mass spectrometry (GC-MS) and enzyme-linked immunosorbent assay (ELISA), respectively. AANAT enzyme activity was measured using liquid biphasic dif-13 fusion assay. Protein levels and mRNA expression of AANAT was 14 measured by Western blot and reverse transcription polymerase 15 chain reaction (RT-PCR) analysis, respectively. Eight hours of nocturnal RFID exposure caused a significant reduction in both urinary melatonin (p = 0. 003) and 6-OHMS (p = 0. 026). Activity, protein levels, and mRNA expression of AANAT were suppressed by exposure to RFID (p < 0. 05). Our results suggest that nocturnal RFID exposure can cause reductions in the levels of both urinary melatonin and 6-OHMS, possibly due to decreased melatonin biosynthesis via suppression of Aanat gene transcription in the rat pineal gland.


"Count on Sleep": an OSA awareness project update.
Obstructive sleep apnea (OSA) is a common, chronic sleep-related breathing disorder that impacts approximately 12% of the United States adult population. Greater public awareness of OSA is necessary to decrease the number of people with undiagnosed or untreated OSA and reduce the negative health consequences of unrecognized OSA. In 2021 the American Academy of Sleep Medicine initiated the "Count on Sleep" project in partnership with key stakeholders with the objective of raising the awareness of OSA among the public, healthcare providers and public health officials about OSA. Four workgroups implemented strategies and completed tasks focused on increasing OSA awareness in their targeted areas to address the objectives of the project including (1) Public Awareness and Communications, (2) Provider Education, (3) Tool Development and Surveillance, and (4) a Strategic Planning workgroup that coordinated efforts across the project. Over the first two years, workgroups made substantiative progress toward project goals including holding "listening sessions" with representatives of communities disproportionately impacted by OSA and its consequences; developing resources for primary care providers that can be easily accessed and used in practice and developing a brief survey for use in estimating and tracking OSA risk across the population. Over the first two project years, workgroups made significant progress in advancing efforts to increase awareness of OSA in US communities. The third year of the project will focus on dissemination of campaign materials and resources for all targeted groups, including the public, healthcare professionals, and public health professionals.


Integrative genetic, genomic and transcriptomic analysis of heat shock protein and nuclear hormone receptor gene associations with spontaneous preterm birth.
Heat shock proteins are involved in the response to stress including activation of the immune response. Elevated circulating heat shock proteins are associated with spontaneous preterm birth (SPTB). Intracellular heat shock proteins act as multifunctional molecular chaperones that regulate activity of nuclear hormone receptors. Since SPTB has a significant genetic predisposition, our objective was to identify genetic and transcriptomic evidence of heat shock proteins and nuclear hormone receptors that may affect risk for SPTB. We investigated all 97 genes encoding members of the heat shock protein families and all 49 genes encoding nuclear hormone receptors for their potential role in SPTB susceptibility. We used multiple genetic and genomic datasets including genome-wide association studies (GWASs), whole-exome sequencing (WES), and placental transcriptomics to identify SPTB predisposing factors from the mother, infant, and placenta. There were multiple associations of heat shock protein and nuclear hormone receptor genes with SPTB. Several orthogonal datasets supported roles for SEC63, HSPA1L, SACS, RORA, and AR in susceptibility to SPTB. We propose that suppression of specific heat shock proteins promotes maintenance of pregnancy, whereas activation of specific heat shock protein mediated signaling may disturb maternal-fetal tolerance and promote labor.


Female mating receptivity after injection of male-derived extracts in Callosobruchus maculatus.
The effects of male-derived extracts on female receptivity were investigated in Callosobruchus maculatus (Coleoptera: Bruchidae). Injection of aqueous extracts of the male reproductive tract into the abdomen of females reduced receptivity. Aqueous extracts of male reproductive tracts were divided to three molecular weight (MW) fractions by ultrafiltration: Fractions: (I) MW<3 kDa, (II) 3-14 kDa, and (III)>14 kDa. Fraction II reduced female receptivity from 3h after injection, and Fraction III reduced female receptivity from 2 days after injection. On the other hand, no effect on receptivity was found for Fraction I. Furthermore, male reproductive tract organs were divided into accessory gland, testis, and seminal vesicle including the ejaculatory duct. Aqueous extracts of the seminal vesicle reduced receptivity of females immediately following injection, while aqueous extracts of the accessory gland reduced receptivity at the second day. The results suggest that the components of Fraction II existed in the seminal vesicle, and those of Fraction III in the accessory gland. The results of the present and the previous studies in Callosobruchus chinensis, a species closely related to C. maculatus, were compared and are discussed from the viewpoint of the significance of ejaculation in the two species.


Orf virus VEGF-E NZ2 promotes paracellular NRP-1/VEGFR-2 coreceptor assembly via the peptide RPPR.
Vascular endothelial growth factors (VEGFs) interact with the receptor tyrosine kinases (RTKs) VEGFR-1, -2, and -3; neuropilins (NRPs); and heparan sulfate (HS) proteoglycans. VEGF RTKs signal to downstream targets upon ligand-induced tyrosine phosphorylation, while NRPs and HS act as coreceptors that lack enzymatic activity yet modulate signal output by VEGF RTKs. VEGFs exist in various isoforms with distinct receptor specificity and biological activity. Here, a series of mammalian VEGF-A splice variants and orf virus VEGF-Es, as well as chimeric and mutant VEGF variants, were characterized to determine the motifs required for binding to NRP-1 in the absence (VEGF-E) or presence (VEGF-A(165)) of an HS-binding sequence. We identified the carboxyterminal peptides RPPR and DKPRR as the NRP-1 binding motifs of VEGF-E and VEGF-A, respectively. RPPR had significantly higher affinity for NRP-1 than DKPRR. VEGFs containing an RPPR motif promoted HS-independent coreceptor complex assembly between VEGFR-2 and NRP-1, independent of whether these receptors were expressed on the same or separate cells grown in cocultures. Functional studies showed that stable coreceptor assembly by VEGF correlated with its ability to promote vessel formation in an embryoid body angiogenesis assay.


Diffusion phenomena of cells and biomolecules in microfluidic devices.
Biomicrofluidics is an emerging field at the cross roads of microfluidics and life sciences which requires intensive research efforts in terms of introducing appropriate designs, production techniques, and analysis. The ultimate goal is to deliver innovative and cost-effective microfluidic devices to biotech, biomedical, and pharmaceutical industries. Therefore, creating an in-depth understanding of the transport phenomena of cells and biomolecules becomes vital and concurrently poses significant challenges. The present article outlines the recent advancements in diffusion phenomena of cells and biomolecules by highlighting transport principles from an engineering perspective, cell responses in microfluidic devices with emphases on diffusion- and flow-based microfluidic gradient platforms, macroscopic and microscopic approaches for investigating the diffusion phenomena of biomolecules, microfluidic platforms for the delivery of these molecules, as well as the state of the art in biological applications of mammalian cell responses and diffusion of biomolecules.


Effects of liposome structure and lipid composition on the activation of the tumoricidal properties of macrophages by liposomes containing muramyl dipeptide.
Various vesicle structures and lipid compositions have been studied to identify the optimal type of liposome for delivery of the macrophage-activating agent muramyl dipeptide (MDP) to macrophages. Evaluation of the ability of liposomes to be phagocytosed by macrophages established that optimal initial rates of engulfment were obtained when multilamellar vesicles (MLV) composed of distearoylphosphatidylcholine (18:0 PC): phosphatidylserine (PS) (7:3 mol ratio) were used. MLV composed exclusively of 18:0 PC were phagocytosed at rates greater than or equal to those of MLV composed of egg phosphatidylcholine (PC):PS, whereas MLV composed only of egg PC were very poorly phagocytosed. Although phagocytosis was enhanced by incorporation of PS into MLV, the inclusion of PS brought about significant enhancement in liposome permeability in the presence of serum. The inclusion of PS, however, was a requirement for the delivery of MLV to the lungs following i.v. injection into mice whether used in conjunction with 18:0 PC or egg PC. Activation of macrophages to become tumoricidal against syngeneic tumor cells wtih liposome-encapsulated MDP was superior in both degree and duration when MLV composed of 18:0 PC:PS (7:3 mol ratio) were used. MLV were found to be superior to large unilamellar vesicles containing equal amounts of lipid and entrapped MDP. On the other hand, higher levels of macrophage activation were obtained when an equivalent amount of a lipophilic MDP derivative, muramyltripeptide:phosphatidylethanolamine, was incorporated into the liposome bilayer irrespective of whether the adjuvant was incorporated in liposomes composed of 18:0 PC:PS or egg PC:PS.


Chromosome mapping of repetitive sequences in Anostomidae species: implications for genomic and sex chromosome evolution.
Members of the Anostomidae family provide an interesting model system for the study of the influence of repetitive elements on genome composition, mainly because they possess numerous heterochromatic segments and a peculiar system of female heterogamety that is restricted to a few species of the Leporinus genus. The aim of this study was to isolate and identify important new repetitive DNA elements in Anostomidae through restriction enzyme digestion, followed by cloning, characterisation and chromosome mapping of this fragment. To identify repetitive elements in other Leporinus species and expand on studies of repetitive elements in Anostomidae, hybridisation experiments were also performed using previously described probes of LeSpeI repetitive elements. The 628-base pair (bp) LeSpeII fragment was hybridised to metaphase cells of L. elongatus individuals as well as those of L. macrocephalus, L. obtusidens, L. striatus, L. lacustris, L. friderici, Schizodon borellii and S. isognathus. In L. elongatus, both male and female cells contained small clusters of LeSpeII repetitive elements dispersed on all of the chromosomes, with enrichment near most of the terminal portions of the chromosomes. In the female sex chromosomes of L. elongatus (Z2,Z2/W1W2), however, this repeated element was absent. In the remaining species, a dispersed pattern of hybridisation was observed on all chromosomes irrespective of whether or not they were sex chromosomes. The repetitive element LeSpeI produced positive hybridisations signals only in L. elongatus, L. macrocephalus and L. obtusidens, i.e., species with differentiated sex chromosomes. In the remaining species, the LeSpeI element did not produce hybridisation signals. Results are discussed in terms of the effects of repetitive sequences on the differentiation of the Anostomidae genome, especially with respect to sex chromosome evolution. LeSpeII showed hybridisation patterns typical of Long Interspersed Elements (LINEs). The differential distribution of this element may be linked to sex chromosome differentiation in L. elongatus species. The relationship between sex chromosome specificity and the LeSpeI element is confirmed in the species L. elongatus, L. macrocephalus and L. obtusidens.


Improving Estrogenic Compound Screening Efficiency by Using Self-Modulating, Continuously Bioluminescent Human Cell Bioreporters Expressing a Synthetic Luciferase.
A synthetic bacterial luciferase-based autobioluminescent bioreporter, HEK293ERE/Gal4-Lux, was developed in a human embryonic kidney (HEK293) cell line for the surveillance of chemicals displaying endocrine disrupting activity. Unlike alternative luminescent reporters, this bioreporter generates bioluminescence autonomously without requiring an external light-activating chemical substrate or cellular destruction. The bioreporter's performance was validated against a library of 76 agonistic and antagonistic estrogenic endocrine disruptor chemicals and demonstrated reproducible half maximal effective concentration (EC50) values meeting the U.S. Environmental Protection Agency (EPA) guidelines for Tier 1 endocrine disrupting chemical screening assays. For model compounds, such as the estrogen receptor (ER) agonist 17β-estradiol, HEK293ERE/Gal4-Lux demonstrated an EC50 value (7.9 × 10-12 M) comparable to that of the current EPA-approved HeLa-9903 firefly luciferase-based estrogen receptor transcription assay (4.6 × 10-12 M). Screening against an expanded array of common ER agonists likewise produced similar relative effect potencies as compared with existing assays. The self-initiated autobioluminescent signal of the bioreporter permitted facile monitoring of the effects of endocrine disrupting chemicals, which decreased the cost and hands-on time required to perform these assays. These characteristics make the HEK293ERE/Gal4-Lux bioreporter potentially suitable as a high-throughput human cell-based assay for screening estrogenic activity.


Symmetrical peripheral gangrene-a case report and brief review.
A 30 year-old gentleman presented to casualty with history of pain abdomen for six days, fever and decreased urine output since two days. He was in a state of septic shock and was diagnosed to have intestinal perforation. His peripheral pulses were not palpable except for the femoral and brachial vessels. Despite fluid resuscitation, he needed infusion of high doses of dopamine and noradrenaline to maintain his blood pressure. He was operated for repair of perforation. On the first postoperative day, in the intensive care unit, vasopressin infusion was added in view of persistent hypotension. Appropriate fluid resuscitation and antibiotic therapy helped to wean him off inotropes and vasopressors by the second postoperative day. On the 3rd postoperative day, however, the patient developed discolouration and blebs on the fingers of left hand, followed by the right hand and then both the lower limbs. Subsequently, over a period of 10 days, this progressed to gangrene formation in the hands despite the patient being haemodynamically stable without any inotropes or vasopressors in this period. We conclude that the septic shock is a systemic derangement affecting all organ systems including coagulation and microcirculation. Early recognition and prompt management of sepsis, optimisation of fluid status to wean off the inotropes and vasopressors at the earliest is necessary to avoid catastrophes such as symmetrical peripheral gangrene.


EMAP, an echinoderm microtubule-associated protein found in microtubule-ribosome complexes.
The major non-tubulin polypeptide found associated with microtubules purified from unfertilized sea urchin eggs by cycles of pH-dependent assembly has a Mr of 77,000. The 77,000 Mr polypeptide is heat- and acid-labile, and is antigenically distinct from the mammalian brain MAPs, MAP-2 and tau. Affinity-purified antiserum against the 77,000 Mr polypeptide was used to survey a variety of cells and tissues for the presence of antigenically related polypeptides. A cross-reacting polypeptide, ranging in Mr from 72,000 to 80,000, was found in microtubule preparations from a wide variety of echinoderms, including sea urchins, starfish and sand dollars. Indirect immunofluorescence showed that the polypetide was found in interphase as well as mitotic microtubule arrays. No cross-reacting material was detected in microtubules isolated from marine molluscs, mammalian brain or mouse B16 cultured cells. Because the 77,000 Mr MAP is abundant in echinoderms, we have called it EMAP for echinoderm microtubule-associated protein. Although the precise function of the EMAP is not known, our data suggest that the EMAP is involved in the attachment of ribosomes to microtubules. Large numbers of ribosomes are attached to the walls of EMAP-containing microtubules, but not EMAP-deficient microtubules. Removal of the EMAP from the microtubule by salt-extraction results in the release of ribosomes from the microtubule, indicating that the EMAP may form part or all of the long tapered stalk that connects these two organelles.


Tetralogy of Fallot.
This article has discussed TOF, its occurrence, physiology, medical interventions and nursing implications. In its classic form, TOF consists of four anatomical aberrations: a large VSD, pulmonic stenosis, dextroposition of the aorta, and right ventricular hypertrophy. Surgical palliation versus primary intracardiac repair is a continuing discussion in medical literature. Children who have undergone intracardiac repair for TOF have an excellent prognosis for late survival, near 90% 10 years after repair. Nursing responsibilities in the care of the newborn diagnosed as having TOF encompass the well-being of the newborn as well as the family. Palliation increases pulmonary arterial flow, thus decreasing cyanosis and promoting measurable clinical improvement in the infant. Preparation of the family and child for surgery involves completing a baseline assessment of family dynamics, diagnosing stressors, composing objectives, carrying out interventions focused on developmental ability, and evaluating the effectiveness of the nursing process. Maintaining the physical and emotional integrity of a child just out of the operating room is a challenge. Parental support is important to the young child's feelings of security while hospitalized.


Reconstruction of human alpha thalassemia-2 genotypes in monkey cells.
The human adult alpha globin genes, alpha 2 and alpha 1, are contained within two tandemly arranged duplication units. Each unit spans 4 kb of DNA, and contains three homology blocks (X, Y, Z) separated by non-homologous sequences. Segmental DNA recombination processes between the two units have resulted in high frequencies of two types of deletions in certain human populations, each deletion removing one alpha globin gene from chromosome 16, (alpha-thalassemia 2). In order to study the molecular mechanisms of alpha-thalassemia 2, and of homologous DNA recombination in general in mammalian cells, we have reconstructed these two alpha-thalassemia 2 genotypes in monkey cells. The two duplication units have been cloned in an SV40 origin-containing vector, and transfected into COS 7 cells. Newly replicated plasmid DNA was isolated and analyzed by Southern blot hybridization. Homologous DNA recombination occurs with high frequencies (10-20% per kb of homology), and this generates both types of alpha-thalassemia 2 deletions on the episomes in the monkey cells. Removal of the 5' end of either one, or both, of the X blocks prior to DNA transfection affects the relative frequencies of the two alpha-thalassemia 2 genotypes in a novel way. We consider and discuss these results in terms of several alternative models. Our data suggest the existence of hot spot(s) for initiation of homologous DNA recombination, or recombination promoting element(s), in a specific region of the human adult alpha globin locus. A DNA sequence that defines the boundaries of the two duplication units, and has been implicated in the initiation of gene conversion of the two X blocks, is contained within this region.


Advances in genomic research on hepatitis C virus with a useful tool, replicon system.
The research for hepatitis C virus (HCV) has long delayed by missing of in vitro culture system. Since the development of replicon system, a replication system of subgenomic HCV RNAs in a hepatoma cell line, has been reported, many virological and clinical findings have been discovered. Recently, in addition of subgenomic replication system, hepatitis C virus full-length RNA replication has been possible, and a few cell culture systems producing viral particles have been produced. These developments enabled us to investigate the life cycle or intracellular circumstance of HCV production. By screening of newly synthesized drugs with this replicon system, several possible medicines have been established and clinical researches are now running. Among them, VX950 and SCH503034 are nearest to clinical use. Other possible agents for reducing viral replication such as cyclophyllin inhibitors, inhibitors of sphingomyelin synthesis, HMG-CoA reductase inhibitors, or RNA-dependent RNA polymerase inhibitors have been also investigated. Furthermore the mechanism for development of hepatocellular carcinoma in the HCV infected liver has been vigorously studied using the HCV replicon system.


Idiopathic thrombocytopenic purpura and MMR vaccine.
A CAUSAL ASSOCIATION BETWEEN MEASLES: mumps-rubella (MMR) vaccine and idiopathic thrombocytopenic purpura (ITP) was confirmed using immunisation/hospital admission record linkage. The absolute risk within six weeks of immunisation was 1 in 22 300 doses, with two of every three cases occurring in the six week post-immunisation period being caused by MMR. Children with ITP before MMR had no vaccine associated recurrences.


Flash Characterization of Smartphones Used in Point-of-Care Diagnostics.
Rapidly growing interest in smartphone cameras as the basis of point-of-need diagnostic and bioanalytical technologies increases the importance of quantitative characterization of phone optical performance under real-world operating conditions. In the context of our development of lateral-flow immunoassays based on phosphorescent nanoparticles, we have developed a suite of tools for characterizing the temporal and spectral profiles of smartphone torch and flash emissions, and their dependence on phone power state. In this work, these tools are described and documented to make them easily available to others, and demonstrated by application to characterization of Apple iPhone 5s, iPhone 6s, iPhone 8, iPhone XR, and Samsung Note8 flash performance as a function of time and wavelength, at a variety of power settings. Flash and torch intensity and duration vary with phone state and among phone models. Flash has high variability when the battery charge is below 10%, thus, smartphone-based Point-of-Care (POC) tests should only be performed at a battery level of at least 15%. Some output variations could substantially affect the results of assays that rely on the smartphone flash.


Reference values for self-recorded blood pressure: a meta-analysis of summary data.
The widespread clinical use of self-recorded blood pressure measurement is limited by the lack of generally accepted reference values. The purpose of this study was therefore to perform a meta-analysis of summary data in an attempt to determine an operational threshold for self-recorded blood pressures. STUDIES AND METHODS: Seventeen studies, including a total of 5422 subjects, were reviewed. Eight of these 17 studies included both normotensive and untreated hypertensive subjects, while the other 9 reports included normotensive subjects only. Within each study an operational cutoff point between normotension and hypertension was derived by means of the mean+2 SDs and the 95th percentiles of the self-recorded blood pressure in normotensive subjects. These 2 methods were contrasted with 2 other techniques that have been applied in the literature to calculate (1) the self-recorded pressures equivalent to a conventional pressure of 140 mm Hg systolic and 90 mm Hg diastolic by means of regression analysis and (2) the self-recorded blood pressures at the percentiles corresponding to a conventional pressure of 140/90 mm Hg. The latter 2 methods were applied in untreated subjects not selected on the basis of their blood pressure. With weighting for the number of subjects included in the various studies, the self-recorded blood pressure averaged 115/71 mm Hg in normotensive persons and 119/74 mm Hg in untreated subjects not selected on the basis of their blood pressure. The reference values for self-recorded blood pressures determined by the mean+2 SDs (137/89 mm Hg) or the 95th percentile (135/86 mm Hg) of the distribution in normotensive subjects were concordant within 2/3 mm Hg, whereas the cutoff points derived with the regression and percentile methods were considerably lower, ie, 125/79 and 129/84 mm Hg, respectively. Until the relationship between self-recorded pressure and the incidence of cardiovascular morbidity and mortality is further clarified by prospective studies, a mean self-recorded blood pressure above 135 mm Hg systolic or 85 mm Hg diastolic may be considered hypertensive.


Comparative Study of Organoids from Patient-Derived Normal and Tumor Colon and Rectal Tissue.
Colon and rectal tumors, often referred to as colorectal cancer, show different gene expression patterns in studies that analyze whole tissue biopsies containing a mix of tumor and non-tumor cells. To better characterize colon and rectal tumors, we investigated the gene expression profile of organoids generated from endoscopic biopsies of rectal tumors and adjacent normal colon and rectum mucosa from therapy-naive rectal cancer patients. We also studied the effect of vitamin D on these organoid types. Gene profiling was performed by RNA-sequencing. Organoids from a normal colon and rectum had a shared gene expression profile that profoundly differed from that of rectal tumor organoids. We identified a group of genes of the biosynthetic machinery as rectal tumor organoid-specific, including those encoding the RNA polymerase II subunits POLR2H and POLR2J. The active vitamin D metabolite 1α,25-dihydroxyvitamin D3/calcitriol upregulated stemness-related genes (LGR5, LRIG1, SMOC2, and MSI1) in normal rectum organoids, while it downregulated differentiation marker genes (TFF2 and MUC2). Normal colon and rectum organoids share similar gene expression patterns and respond similarly to calcitriol. Rectal tumor organoids display distinct and heterogeneous gene expression profiles, with differences with respect to those of colon tumor organoids, and respond differently to calcitriol than normal rectum organoids.


Weight increase in people with cystic fibrosis on CFTR modulator therapy is mainly due to increase in fat mass.
Background: Ivacaftor, the first CFTR modulator drug, leads to significant long-term improvement in lung function and weight gain. The mechanism as well as the long-term impact of ivacaftor on weight, resting energy expenditure (REE) and body composition remains to be explored. Methods: This prospective observational study included 18 people with CF (pwCF) (age: median (range) 20 (6-58) years) carrying at least one CFTR gating mutation commencing ivacaftor. Assessments of body composition, REE and laboratory investigations were performed at baseline and 6, 12 and 24 months after treatment initiation. Results: Treatment with ivacaftor was associated with a significantly positive change in BMI z-score at 24 months. Fat mass (mean (95% CL) of 6.5 kg (4.0; 9.0) from baseline, p = 0.0001), but not fat-free mass changed under ivacaftor treatment. There was a significant positive correlation between weight and fat mass change. Overall, there was no significant change in measured REE from baseline (mean (95% CL) of 108 kcal/d (-12; 228), p = 0.07) in our cohort. Pancreatic function and other nutritional markers did not change with treatment, with the exception of an increase in serum vitamin A levels (p = 0.006). Conclusion: The weight gain observed in ivacaftor treated pwCF is predominantly secondary to increases in fat mass warranting early counseling of people starting on CFTR-modulating treatment with respect to healthy diet and physical exercise.


Randomized phase II study of gemcitabine plus S-1 combination therapy vs. S-1 in advanced biliary tract cancer: Japan Clinical Oncology Group Study (JCOG0805).
A randomized Phase II selection design trial comparing gemcitabine plus S-1 combination therapy with S-1 monotherapy for chemo-naïve unresectable or recurrent biliary tract cancer patients was started in Japan. The aim of this trial is to evaluate the efficacy and safety of the two regimens and to determine which is more promising as a test arm regimen to be compared with the current standard regimen, gemcitabine plus cisplatin, in a subsequent Phase III trial. Patients with unresectable or recurrent biliary tract cancer are randomized to either gemcitabine plus S-1 combination therapy arm or S-1 monotherapy arm. A total of 100 patients will be accrued for this study from 18 institutions over 1 year. The primary endpoint is the proportion of 1-year overall survival, and the secondary endpoints are progression-free survival, response rate and adverse events.


Adaptive denoising and multiscale detection of the V wave in brainstem auditory evoked potentials.
This paper describes a wavelet-transform-based system for the V wave identification in brainstem auditory evoked potentials (BAEP). The system combines signal denoising and rule-based localization modules. The signal denoising module has the potential of effective noise reduction after signal averaging. It analyses adaptively the evolution of the wavelet transform maxima across scales. The singularities of the signal create wavelet maxima with different properties from those of the induced noise. A non-linear filtering process implemented with a neural network extracts out the noise-induced maxima. The filtered wavelet details are subsequently analysed by the rule-based localization module for the automatic identification of the V wave. In the first phase, it implements a set of statistical observations as well as heuristic criteria used by human experts in order to classify the IV-V complex. At the second phase, using a multiscale focusing algorithm, the IV and V waves are positioned on the BAEP signal. Our experiments revealed that the system provides accurate results even for signals exhibiting unclear IV-V complexes.


Synergistic cytotoxicity of pyrazoloacridine with doxorubicin, etoposide, and topotecan in drug-resistant tumor cells.
Pyrazoloacridine (NSC 366140, PD115934, PZA) is a new class of acridine anticancer agents under investigation in Phase II clinical trials in patients with advanced cancers. Although poor responses in patients to the treatment with PZA alone have been observed, this class of agents remains of interest because of its distinct mechanism of action from other topoisomerase poisons. Therefore, the combination of PZA with conventional anticancer agents presents an attractive approach to treat drug-resistant human tumors. In the present study, the cytotoxic effects of PZA combined with doxorubicin, topotecan, and etoposide were determined using paired parental and doxorubicin-resistant human colon carcinoma (SW-620 and SW620/AD-300) and breast cancer cell lines (MCF-7 and MCF-7/TH). Cytotoxicity was measured by soft agar clonogenic assays. Dose effect and combination effects were analyzed by the method of Chou and Talalay. The combination of PZA with doxorubicin, topotecan, and etoposide in fixed ratios demonstrated synergistic cytotoxicity on both SW-620 and SW620/AD-300 cell lines. The combination of PZA with doxorubicin also exhibited synergistic cytotoxicity against both MCF-7 and MCF-7/TH cell lines. The mechanism of synergism appeared independent of topoisomerase I and II inhibition, and interference with protein-DNA complexes. Strategies to define optimal drug combinations are proving to be of significant value when considering potential clinical applications of new and established agents.


MetaOptics: opensource software for designing metasurface optical element GDSII layouts.
Metasurfaces have recently emerged as a promising technology to realize flat and ultra-thin optical elements that can manipulate light at sub-wavelength scale. The typical design flow of a metasurface involves tedious Finite Difference Time Domain (FDTD) simulations followed by creation of a GDSII layout of the metasurface phase profile, the latter being essential for fabrication purposes. Both these steps can be time-consuming and involve the usage of expensive software. To make the design process more straightforward, we have developed an open-source software called MetaOptics built using Python for designing a generic metasurface optical element. MetaOptics uses the FDTD simulated phase response data of a set of meta-atoms and converts the phase profile of any given optical element into a metasurface GDSII layout. MetaOptics comes with in-built FDTD data for most commonly used wavelengths in the visible and infrared spectrum. It also has an option to upload user-specific dimension versus transmission phase data for any choice of wavelength. In this work we describe the software's framework and provide details to guide users to design a metasurface layout using MetaOptics.


Agl16, a thermophilic glycosyltransferase mediating the last step of N-Glycan biosynthesis in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius.
Recently, the S-layer protein of Sulfolobus acidocaldarius was shown to be N-linked with a tribranched hexasaccharide, composed of Man2Glc1GlcNAc2 and a sulfated sugar called sulfoquinovose. To identify genes involved in the biosynthesis and attachment of this glycan, markerless in-frame deletions of genes coding for predicted glycosyltransferases were created. The successful deletion of agl16, coding for a glycosyltransferase, resulted in the S-layer protein and archaellins having reduced molecular weights, as visualized by Coomassie staining or immunoblotting. This analysis indicated a change in the N-glycan composition. Nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses confirmed that the glycan of the S-layer protein from the agl16 deletion mutant was a pentasaccharide, which was missing a terminal hexose residue. High-performance liquid chromatography (HPLC) analyses of the hydrolyzed N-glycan indicated that the missing hexose is a glucose residue. A physiological characterization of the agl16 deletion mutant revealed a significant effect on the growth at elevated salt concentrations. At 300 mM NaCl, the doubling time of the Δagl16 mutant was increased 2-fold compared to that of the background strain. Furthermore, the incomplete glycan structure of the Δagl16 deletion strain affected the assembly and function of the archaellum, as exemplified by semisolid Gelrite plate analysis, in which the motility is decreased according to the N-glycan size.


A genetic locus involved in iron utilization unique to some Campylobacter strains.
Two genes involved in iron utilization in Campylobacter coli VC167 T1 have been characterized. The cfrA gene encodes a protein with a predicted Mr of 77,653 which, after processing of the leader sequence, has a predicted Mr of 75,635. This protein has significant sequence identity to siderophore receptors of several bacteria, and site-specific mutants defective in cfrA do not synthesize one of two major iron-repressible outer membrane proteins. An adjacent gene encodes a TonB-like protein; a mutant in this gene lost the ability to utilize hemin, ferrichrome, and enterochelin as iron sources. The cfrA and tonB genes of VC167 T1 hybridized to all strains of C. coli and most strains of C. jejuni examined but did not hybridize to several other strains of C. jejuni, suggesting that the thermophilic campylobacters can be separated into two categories based on the presence of these two iron utilization genes.


Effects of vascular flow and PEEP in a multiple hit model of lung injury in isolated perfused rabbit lungs.
High vascular flow aggravates lung damage in animal models of ventilator-induced lung injury. Positive end-expiratory pressure (PEEP) can attenuate ventilator-induced lung injury, but its continued effectiveness in the setting of antecedent lung injury is unclear. The objective of the present study was to evaluate whether the application of PEEP diminishes lung injury induced by concurrent high vascular flow and high alveolar pressures in normal lungs and in a preinjury lung model. Two series of experiments were performed. Fifteen sets of isolated rabbit lungs were randomized into three groups (n = 5): low vascular flow/low PEEP; high vascular flow/low PEEP, and high vascular flow/high PEEP. Subsequently, the same protocol was applied in an additional 15 sets of isolated rabbit lungs in which oleic acid was added to the vascular perfusate to produce mild to moderate lung injury. All lungs were ventilated with peak airway pressure of 30 cm H2O for 30 minutes. Outcome measures included frequency of gross structural failure, pulmonary hemorrhage, edema formation, changes in static compliance, pulmonary vascular resistance, and pulmonary ultrafiltration coefficient. In the context of high vascular flow, application of a moderate level of PEEP reduced pulmonary rupture, edema formation, and lung hemorrhage. The protective effects of PEEP were not observed in lungs concurrently injured with oleic acid. Under these experimental conditions, PEEP attenuates lung injury in the setting of high vascular flow. The protective effect of PEEP is lost in a two-hit model of lung injury.


Importin-β facilitates nuclear import of human GW proteins and balances cytoplasmic gene silencing protein levels.
MicroRNAs (miRNAs) guide Argonaute (Ago) proteins to distinct target mRNAs leading to translational repression and mRNA decay. Ago proteins interact with a member of the GW protein family, referred to as TNRC6A-C in mammals, which coordinate downstream gene-silencing processes. The cytoplasmic functions of TNRC6 and Ago proteins are reasonably well established. Both protein families are found in the nucleus as well. Their detailed nuclear functions, however, remain elusive. Furthermore, it is not clear which import routes Ago and TNRC6 proteins take into the nucleus. Using different nuclear transport assays, we find that Ago as well as TNRC6 proteins shuttle between the cytoplasm and the nucleus. While import receptors might function redundantly to transport Ago2, we demonstrate that TNRC6 proteins are imported by the Importin-β pathway. Finally, we show that nuclear localization of both Ago2 and TNRC6 proteins can depend on each other suggesting actively balanced cytoplasmic Ago - TNRC6 levels.


The use of genetic typing methods to discriminate among strains of Vibrio cholerae, V. parahaemolyticus, and V. vulnificus.
This review article summarizes the findings of recent typing studies conducted on Vibrio cholerae, V. parahaemolyticus, and V. vulnificus. The DNA-based methods used to type the Vibrio spp. include whole genome approaches, such as pulsed field gel electrophoresis (PFGE), ribotyping, and repetitive extragenic palindromic (REP)-PCR, single gene targets, and multiple gene targets (multilocus approaches). The goals of these studies include establishing the relatedness of isolates from disease epidemics, discriminating among strains with more or less potential to cause disease or epidemics, and exploring the population biology of these waterborne pathogens. PFGE was consistently among the more discriminatory of the typing methods for all three Vibrio spp., and was useful for tracing the temporal and geographic relatedness of epidemic strains of V. cholerae and V. parahaemolyticus. However, PFGE did not group V. vulnificus strains according to the genotypes that have been proposed as markers of virulence potential. Typing methods that target repetitive elements distributed throughout the genome, such as BOX-PCR and REP-PCR, and DNA sequence-based methods, such as multilocus sequence typing, were also highly discriminatory and, in some cases, superior to PFGE for phylogenetic analysis and identification of strains with high epidemic or virulence potential. As typing methods and strategies are refined and used, the epidemiology, virulence potential, and ecology of these pathogenic Vibrio spp. will become better understood.


The contribution of cervical dynamic magnetic resonance imaging to the surgical treatment of cervical spondylotic myelopathy.
Cervical spine is the most kinetic segment of the whole vertebrae. The radiologic imaging methods concern with the morphologic changes but give no functional data. At flexion, spinal cord strains, anterior osteophytic compression increases. At extension, spinal canal gets narrower, cord shortens and gets thicker, compression of posterior ligament gets abberant and cord compression increases. 258 cervical spondylotic myelopathy (CSM) cases were scanned by conventional magnetic resonance imaging (MRI) and additionally dynamic MRI. Contributions of dynamic cervical MRI to the surgical plans and results were evaluated. We had sagittal and axial T2W MR scans at flexion and extension, in addition to the neutral cervical MR imaging. We found that the AP diameter of spinal canal is increased 14.9 % in flexion and decreased 13.4 % in extension relative to the neutral MR imaging. The changes of the cord compression and the transvers area of cord which is the most important prognostic indicator in spinal diseases and also the area of spinal cord and subarachnoid space can be detected via dynamic axial sections of MRI. Dynamic MR images may be helpfull in the decision making for the surgical treatment of CSM.


Transcript Dynamics in Wounded and Inoculated Scots Pine.
Comparative transcriptome analysis provides a useful tool for the exploration of plant-pathogen interaction by allowing in-depth comparison of gene expression between unaffected, inoculated and wounded organisms. Here we present the results of comparative transcriptome analysis in genetically identical one-year-old Scots pine ramets after wounding and inoculation with Heterobasidion annosum. We identified 230 genes that were more than 2-fold upregulated in inoculated samples (compared to controls) and 116 downregulated genes. Comparison of inoculated samp les with wounded samples identified 32 differentially expressed genes (30 were upregulated after inoculation). Several of the genes upregulated after inoculation are involved in protection from oxidative stress, while genes involved in photosynthesis, water transport and drought stress tolerance were downregulated. An NRT3 family protein was the most upregulated transcript in response to both inoculation and wounding, while a U-box domain-containing protein gene was the most upregulated gene comparing inoculation to wounding. The observed transcriptome dynamics suggest involvement of auxin, ethylene, jasmonate, gibberellin and reactive oxygen species pathways and cell wall modification regulation in response to H. annosum infection. The results are compared to methyl jasmonate induced transcriptome dynamics.


Feedback-regulated poly(ADP-ribosyl)ation by PARP-1 is required for rapid response to DNA damage in living cells.
Genome integrity is constantly threatened by DNA lesions arising from numerous exogenous and endogenous sources. Survival depends on immediate recognition of these lesions and rapid recruitment of repair factors. Using laser microirradiation and live cell microscopy we found that the DNA-damage dependent poly(ADP-ribose) polymerases (PARP) PARP-1 and PARP-2 are recruited to DNA damage sites, however, with different kinetics and roles. With specific PARP inhibitors and mutations, we could show that the initial recruitment of PARP-1 is mediated by the DNA-binding domain. PARP-1 activation and localized poly(ADP-ribose) synthesis then generates binding sites for a second wave of PARP-1 recruitment and for the rapid accumulation of the loading platform XRCC1 at repair sites. Further PARP-1 poly(ADP-ribosyl)ation eventually initiates the release of PARP-1. We conclude that feedback regulated recruitment of PARP-1 and concomitant local poly(ADP-ribosyl)ation at DNA lesions amplifies a signal for rapid recruitment of repair factors enabling efficient restoration of genome integrity.


The benefit of the doubt or doubts over benefits? A systematic literature review of perceived risks of vaccines in European populations.
The success of vaccination strategies depends in part on population perceptions of benefits and risks of vaccines and related confidence in vaccination. Better knowledge of public concerns about vaccines and what is driving them is needed to inform vaccination strategies and communications. This literature reviewer examined studies on vaccine and vaccination risk perceptions and concerns across European populations. A systematic literature review was conducted to identify studies published between 2004 and 2014 in Europe. A descriptive analysis was performed. A total of 145 articles were selected, most of which were conducted in the UK, the Netherlands and France and studied seasonal influenza, HPV and pandemic influenza vaccination. Across all countries and vaccines, the primary area of concern was vaccine safety, followed by perceptions of low likelihood of contracting vaccine-preventable diseases (VPDs), perceived low severity of VPDs, beliefs that vaccines do not work, and overall lack of information. Concerns were found to be vaccine-, country- and population-specific. In addition to identifying concerns about vaccination in Europe, this study confirmed the notion that individuals have many safety concerns about vaccination and often believe that the risks of vaccination outweigh their benefits. More research needs to be conducted to explore the impact of different types of communication strategies, which would frame the benefits of vaccination as well as risks of not vaccinating. Strategies to better inform public perceptions of vaccines should include the provision of unbiased, comprehensive information tailored to population information needs, and delivered using multiple and new communication technologies such as social media.