Toward a culture-by-context perspective on negotiation: negotiating teams in the United States and Taiwan.
Within the United States, teams outperform solos in negotiation (Thompson, Peterson, & Brodt, 1996). The current research examined whether this team advantage generalizes to negotiators from a collectivist culture (Taiwan). Because different cultures have different social norms, and because the team context may amplify the norms that are salient in a particular culture (Gelfand & Realo, 1999), we predicted that the effect of teams on negotiation would differ across cultures. Specifically, we predicted that since harmony norms predominate in collectivist cultures like Taiwan, the team context would amplify a concern with harmony, leading Taiwanese teams to negotiate especially suboptimal outcomes. In support, 2 studies showed that Taiwanese teams negotiated less-optimal outcomes than Taiwanese solos. We also used a moderated-mediation analysis to investigate the mechanism (Hayes, 2012), documenting that the interactive effect of culture and context on outcomes was mediated by harmony norms. By showing that the same situational conditions (team negotiations) can have divergent effects on negotiation outcomes across cultures, our results point toward a nuanced, sociocontextual view that moves beyond the culture-as-main-effect approach to studying culture and negotiations.


The Influence of the Intraductal Carcinoma of the Prostate on the Short-Term Oncological Outcomes.
The presence of intraductal carcinoma of the prostate (IDC-P) in radical prostatectomy (RP) specimens correlates with adverse prognostic factors such as worse biochemical recurrence-free survival, higher grade and stage disease. This study aimed to investigate the effect of IDC-P in radical prostatectomy specimens on short-term oncological outcomes. Patients who underwent RP at our clinic for prostate cancer between May 2016 and November 2019 were included in the study. They were divided into two groups based on the presence of IDC-P in RP specimens. Their clinical, pathological, and oncologic data were evaluated retrospectively. A total of 98 patients underwent RP with a mean age of 65.5 years (50-83) and a mean follow-up time of 31.2 months (6-52). Seventy and 28 patients were evaluated in the group without IDC-P and group with IDC-P, respectively. Surgical margin positivity (p=0.307) and lymph node metastasis (p=0.017) rates were higher in the group with IDC-P. Although there were no statistical differences between the groups, at follow-up biochemical recurrence rate (p=0.052) was higher, and mean time to biochemical recurrence rates were lower (p=0.057) in the group with IDC-P. The group with IDC-P was associated with a 3-fold increase in prostate cancer-specific mortality to the group without IDC-P (p=0.037). Patients with IDC-P at RP specimens have more advanced disease, shorter biochemical recurrence-free, and cancerspecific survival than those without IDC-P. Defining the presence of IDC-P in RP specimens is critical in choosing the appropriate treatment strategy and predicting the prognosis.


Stretchable and transparent electrodes using hybrid structures of graphene-metal nanotrough networks with high performances and ultimate uniformity.
Transparent electrodes that can maintain their electrical and optical properties stably against large mechanical deformations are essential in numerous applications of flexible and wearable electronics. In this paper, we report a comprehensive analysis of the electrical, optical, and mechanical properties of hybrid nanostructures based on graphene and metal nanotrough networks as stretchable and transparent electrodes. Compared to the single material of graphene or the nanotrough, the formation of this hybrid can improve the uniformity of sheet resistance significantly, that is, a very low sheet resistance (1 Ω/sq) with a standard deviation of less than ±0.1 Ω/sq, high transparency (91% in the visible light regime), and superb stretchability (80% in tensile strain). The successful demonstration of skin-attachable, flexible, and transparent arrays of oxide semiconductor transistors fabricated using hybrid electrodes suggests substantial promise for the next generation of electronic devices.


Synapsins as major neuronal Ca2+/S100A1-interacting proteins.
The mammalian S100A1 protein can activate the invertebrate myosin-associated giant protein kinase twitchin in a Ca(2+)-dependent manner by more than 1000-fold in vitro; however, no mammalian S100-dependent protein kinases are known. In an attempt to identify novel mammalian Ca(2+)/S100A1-regulated protein kinases, brain extracts were subjected to combined Ca(2+)-dependent affinity chromatography with S100A1 and an ATP analogue. This resulted in the purification to near-homogeneity of the four major synapsin isoforms Ia, Ib, IIa and IIb. All four synapsins were specifically affinity-labelled with the ATP analogue 5'-p-fluorosulphonylbenzoyladenosine. S100A1 bound to immobilized synapsin IIa in BIAcore experiments in a Ca(2+)-dependent and Zn(2+)-enhanced manner with submicromolar affinity; this interaction could be competed for with synthetic peptides of the proposed S100A1-binding sites of synapsin. Double-labelling confocal immunofluorescence microscopy demonstrated that synapsins and S100A1 are both present in the soma and neurites of PC12 cells, indicating their potential to interact in neurons in vivo.


Influence of Urban Green Space and Facility Accessibility on Exercise and Healthy Diet in Hong Kong.
Background A cross-sectional study using a convenience sampling method was conducted to understand how green space and accessibility of common public open spaces in compact urban areas affect physical activity and healthy diets of residents. Methods A total of 554 residents completed a structured questionnaire on quality of life, physical activity level and healthy eating practice. Particularly, categories of physical activity and durations were obtained by using the short form Chinese International Physical Activity Questionnaire (IPAQ-C), then the Metabolic Equivalent of Task (MET)-minutes/week was calculated using the formulae (walking minutes × walking days × 3.3) + (moderate-intensity activity minutes × moderate days × 4.0) + (vigorous-intensity activity minutes × vigorous-intensity days × 8.0). The percentage of green space was calculated based on a spatial buffer with a 500 m radius from participants' geocoded addresses using a SPOT ('Satellite Pour l'Observation de la Terre' in French) satellite image-derived vegetation dataset. Parks, promenade and sports facilities were examples of open spaces. Results The sampled population who lived with green space averaged 10.11% ± 7.95% (ranged 1.56-32.90%), with the majority (90%) performing physical activities at medium and high levels. MET-minutes/week was significantly associated (Pearson r = 0.092; p < 0.05) with the green space percentage. Relatively active residents commonly used open spaces within the district for performing exercise, in particular, parks and promenades were mostly used by older residents, while sports facilities by the younger groups at age 25-44 and <25 years. Conclusions Current findings suggested promotion of exercise could be achieved by the design or redesign of built environment to include more parks accessible to the residents with the increase of vegetation.


Comparison of Neck Injury Criteria Values Across Human Body Models of Varying Complexity.
Due to the severity and frequency of cervical spine injuries, the neck injury criterion (Nij) was developed to provide a quantitative relationship between forces and moments of the upper neck with accompanied injury risk. The present study was undertaken to evaluate differences in calculated Nij for the Global Human Body Model Consortium's detailed and simplified average 50th percentile male models. The simplified model is a computationally light version of the more detailed model and therefore it is of interest to achieve similar Nij values between the two models. These models were compared in 15 match paired conditions of rigid head impact and a mixture of seven full body rigid hub and sled pulses, for 44 total simulations. Collectively, Nij values of the simplified model were found to exhibit a second-degree polynomial fit, allowing for a conversion to the prediction of the detailed model. Correlates were also derived for impact and inertial loading cases individually, for which the latter may be the subject of future work. The differences in Nij may be attributed to a variety of modeling approach differences related to neck muscles (attachment location and morphometric implementation), localization of head mass within the M50-OS, head geometry, and intervertebral joint space properties. With a primary focus on configurations in the anterior-posterior direction, there is a potential limitation in extensibility to lateral loading cases. In response to the relatively low Nij values exhibited, future work should evaluate the appropriateness of the established critical intercepts of Nij for computational human body models.


Parameter estimation following an adaptive treatment selection trial design.
Two-stage, drop-the-losers designs for adaptive treatment selection have been considered by many authors. The distributions of conditional sufficient statistics and the Rao-Blackwell technique were used to obtain an unbiased estimate and to construct an exact confidence interval for the parameter of interest. In this paper, we characterize the selection process from a binomial drop-the-losers design using a truncated binomial distribution. We propose a new estimator and show that it is asymptotically consistent with a large sample size in either the first stage or the second stage. Supported by simulation analyses, we recommend the new estimator over the naive estimator and the Rao-Blackwell-type estimator based on its robustness in the finite-sample setting. We frame the concept as a simple and easily implemented procedure for phase 2 oncology trial design that can be confirmatory in nature, and we use an example to illustrate its application.


Optical Control of G-Actin with a Photoswitchable Latrunculin.
Actin is one of the most abundant proteins in eukaryotic cells and a key component of the cytoskeleton. A range of small molecules have emerged that interfere with actin dynamics by either binding to polymeric F-actin or monomeric G-actin to stabilize or destabilize filaments or prevent their formation and growth, respectively. Amongst these, the latrunculins, which bind to G-actin and affect polymerization, are widely used as tools to investigate actin-dependent cellular processes. Here, we report a photoswitchable version of latrunculin, termed opto-latrunculin (OptoLat), which binds to G-actin in a light-dependent fashion and affords optical control over actin polymerization. OptoLat can be activated with 390 - 490 nm pulsed light and rapidly relaxes to the inactive form in the dark. Light activated OptoLat induced depolymerization of F-actin networks in oligodendrocytes and budding yeast, as shown by fluorescence microscopy. Subcellular control of actin dynamics in human cancer cell lines was demonstrated by live cell imaging. Light-activated OptoLat also reduced microglia surveillance in organotypic mouse brain slices while ramification was not affected. Incubation in the dark did not alter the structural and functional integrity of microglia. Together, our data demonstrate that OptoLat is a useful tool for the elucidation of G-actin dependent dynamic processes in cells and tissues.


[Regulation of pancreatic secretion by negative feedback and blood gastrointestinal hormones in the pig].
The aim of this study was to determine the rate of circulating gastrointestinal hormones in pigs when pancreatic secretion was re-introduced or removed from the duodenal lumen. Permanent fistulas were fitted into the pancreatic duct and duodenum of 16 pigs, and catheters were introduced into the portal vein and a jugular vein as well. Plasma hormone content was determined at different times during two periods of 30 and 60 min each, when the juice was returned to the pigs; these two periods included a 120-min interval when the juice was not reintroduced into the intestinal lumen. When the pancreatic juice was not returned to the pigs, plasma secretin content rose significantly in the portal blood (+ 42.8% at min 15; +52.5% at min 30; +31% at min 60 and +27% at min 120) and in the peripheral blood (+36.4% at min 30; +18.9% at min 120) compared to the mean values recorded during intraduodenal re-introduction of the secretion. In parallel, the volume of juice secreted and protein output increased significantly but their concentration did not. When the pancreatic juice was removed the mean cholecystokinin level in the portal vein did not change significantly, except at min 30 when its value was 32% higher than the mean during the periods of restitution. Cholecystokinin in the peripheral blood was not affected by removing the pancreatic juice from the intestinal lumen. The mean plasma levels of gastrin, somatostatin, VIP and PP in the portal and peripheric blood were unaltered. These results suggest that in pigs secretin may have an important role in the regulation of pancreatic secretion by negative feedback.


Benefits and barriers to consumer choice.
Any successful health care reform effort must increase the market power of individual consumers. If consumers act in the medical marketplace as they do in other segments of the economy they will make sensible decisions about the allocation of resources (rationing) without the intrusion of third parties and will select providers and medical interventions that meet their needs. This article asserts the importance of consumer influence, focuses on the barriers that prevent consumers from exerting the same muscle in health care that they manifest in other areas of the economy, and suggests ways to remove these barriers.


Developing standardized corticosteroid treatment for Duchenne muscular dystrophy.
Despite corticosteroids being the only treatment documented to improve strength and function in boys with Duchenne muscular dystrophy (DMD) corticosteroid prescription is inconsistent and in some countries, corticosteroids are not prescribed. We are conducting a clinical trial that (1) compares the 3 most frequently prescribed corticosteroid regimes; (2) standardizes treatment of DMD complications; and (3) standardizes prevention of corticosteroid side effects. Investigators at 38 sites in 5 countries plan to recruit 300 boys aged 4-7 who are randomly assigned to one of three regimens: daily prednisone; daily deflazacort; or intermittent prednisone (10days on/10days off). Boys are followed for a minimum of 3years to assess the relative effectiveness and adverse event profiles of the different regimens. The primary outcome is a 3-dimensional variable consisting of log-transformed time to rise from the floor, forced vital capacity, and subject/parent satisfaction with treatment, each averaged over all post-baseline visits. The study protocol includes evidence- and consensus-based treatment of DMD complications and of corticosteroid side effects. This study seeks to establish a standard corticosteroid regimen for DMD. Since all new interventions for DMD are being developed as add-on therapies to corticosteroids, defining the optimum regimen is of importance for all new treatments.


Nuclear transcription factor κ B activation and protein turnover adaptations in skeletal muscle of patients with progressive stages of lung cancer cachexia.
Experimental models of cancer cachexia have indicated that systemic inflammation induces muscle-protein breakdown and wasting via muscular nuclear transcription factor κB (NF-κB) activation. This process may limit the efficacy of nutritional intervention. We assessed muscle NF-κB activity and protein turnover signaling in progressive stages of clinical lung cancer cachexia and assessed whether circulating factors can induce muscular NF-κB activity. Patients with lung cancer precachexia (n = 10) and cachexia (n = 16) were cross-sectionally compared with 22 healthy control subjects. mRNA transcripts of muscle proteolytic (ubiquitin proteasome system and autophagy lysosomal pathway) and myogenic markers and protein expression of PI3K/Akt, myostatin, and autophagy signaling were measured. A multiplex analysis showed the systemic inflammatory status, whereas plasma exposure to stable NF-κB-luciferase-reporter muscle cells revealed NF-κB inducibility. Compared with healthy control subjects, cachectic patients had reduced (appendicular) muscle mass (-10%), muscle fiber atrophy (-27%), and decreased quadriceps strength (-31%). Subtle alterations in the muscle morphology were also detectable in precachectic patients, without changes in body composition. Despite increased Akt phosphorylation, downstream phosphosubstrates glycogen synthase kinase 3β, mammalian target of rapamycin, and Forkhead box protein were unaltered. The expression of autophagy effectors B cell lymphoma 2/adenovirus E1B 19-kDa protein-interacting protein 3 and microtubule-associated proteins 1A/1B light chain 3B gradually increased from precachectic to cachectic patients, without differences in E3 ubiquitin ligases. Systemic and local inflammation was evident in cachexia and intermediate in precachexia, but the plasma of both patients groups caused ex vivo muscle NF-κB activation. In lung cancer, muscular NF-κB activity is induced by factors contained within the circulation. Autophagy may contribute to increased muscle proteolysis in lung cancer cachexia, whereas the absence of downstream changes in phosphosubstrates despite increased Akt phosphorylation suggests impaired anabolic signaling that may require targeted nutritional intervention.


Functional recovery after cesarean delivery: a prospective cohort study in rural Rwanda.
Women who deliver via cesarean section (c-section) experience short- and long-term disability that may affect their physical health and their ability to function normally. While clinical complications are assessed, postpartum functional outcomes are not well understood from a patient's perspective or well-characterized by previous studies. In Rwanda, 11% of rural women deliver via c-section. This study explores the functional recovery of rural Rwandan women after c-section and assesses factors that predict poor functionality at postoperative day (POD) 30. Data were collected prospectively on POD 3, 11, and 30 from women delivering at Kirehe District Hospital between October 2019 and March 2020. Functionality was measured by self-reported overall health, energy level, mobility, self-care ability, and ability to perform usual activities; and each domain was rated on a 4-point likert scale, lower scores reflecting higher level of difficulties. Using the four functionality domains, we computed composite mean scores with a maximum score of 4.0 and we defined poor functionality as composite score of ≤ 2.0. We assessed functionality with descriptive statistics and logistic regression. Of 617 patients, 54.0%, 25.9%, and 26.8% reported poor functional status at POD3, POD11, and POD30, respectively. At POD30, the most self-reported poor functionality dimensions were poor or very poor overall health (48.1%), and inability to perform usual activities (15.6%). In the adjusted model, women whose surgery lasted 30-45 min had higher odds of poor functionality (aOR = 1.85, p = 0.01), as did women who experienced intraoperative complications (aOR = 4.12, 95% CI (1.09, 25.57), p = 0.037). High income patients had incrementally lower significant odds of poor physical functionality (aOR = 0.62 for every US$1 increase in monthly income, 95% CI (0.40, 0.96) p = 0.04). We found a high proportion of poor physical functionality 30 days post-c-section in this Rwandan cohort. Surgery lasting > 30 min and intra-operative complications were associated with poor functionality, whereas a reported higher income status was associated with lower odds of poor functionality. Functional status assessments, monitoring and support should be included in post-partum care for women who delivered via c-section. Effective risk mitigating intervention should be implemented to recover functionality after c-section, particularly among low-income women and those undergoing longer surgical procedures or those with intraoperative complications.


Hypophosphataemic osteomalacia in fibrous dysplasia.
We describe three patients with fibrous dysplasia of bone in whom there was evidence of hypophosphataemic osteomalacia or rickets. Two of the patients had polyostotic fibrous dysplasia and osteomalacia. The third was a child with fibrous dysplasia of the facial and cranial bones and rickets. In all cases the manifestations of osteomalacia or rickets were controlled with large doses of vitamin D. In the child the rickets and hypophosphataemia ceased when most of the bone affected by fibrous dysplasia was surgically resected. Previously reported cases of the association between fibrous dysplasia and hypophosphataemic osteomalacia are reviewed. We suggest that these cases are analogous to the syndrome of 'tumour rickets' where hypophosphataemia appears to be due to the presence of a mesenchymal tumour and regresses when the tumour is removed.


Unaltered norepinephrine-heart rate relationship in exercise with exogenous heat.
We measured plasma norepinephrine (NE) concentration, an index of sympathetic nervous activity, and epinephrine (E), an index of adrenal medulla activity, in six normal young men during mild to severe exercise, with and without superimposed heat stress. The primary objective was to observe whether the normally close relationship between heart rate and log NE concentration in upset when heart rate at a given work load is increased by heat stress. Exercise, beginning at 50 W, was graded in 50-W increments lasting 10 min each up to 200 W, which lasted 5-10 min. Each subject went through the protocol twice, once with skin temperature kept low by a water-perfused suit and then with skin temperature raised to 38 degrees C. Exogenous heart stress raised log circulating NE concentration in proportion to the rise in heart rate at a given work load so that the usual relationship between these variables, previously observed during other stresses, was preserved. In contrast to some other stresses, heat stress had no added effect on E concentration, indicating that this stress during exercise raises sympathetic neural activity (as reflected in the rise in NE) without stimulating additional adrenal release of E.


Production of sterigmatocystin by some species of the genus Aspergillus and its toxicity to chicken embryos.
Sterigmatocystin was produced by 59% of Aspergillus flavus cultures and by 16% of A. parasiticus cultures. All sterigmatocystin-producing cultures of the A. flavus group also simultaneously produced aflatoxin or O-methylsterigmatocystin. Sterigmatocystin was produced by A. chevalieri, A. ruber, and A. amstelodami, species not previously reported to produce the compound. In 5-day-old chicken embryos, the no-effect level of toxicity of sterigmatocystin was between 1 and 2 mug/egg; the mean lethal dose was 5 to 7 mug; and 90 to 100% of the embryos were killed with 10 mug. Teratogenic effects and weight reduction were generally associated with nonlethal doses.


Interacting regulatory circuits involved in orderly control of photosynthesis gene expression in Rhodobacter sphaeroides 2.4.1.
FnrL, the homolog of the global anaerobic regulator Fnr, is required for the induction of the photosynthetic apparatus in Rhodobacter sphaeroides 2.4.1. Thus, the precise role of FnrL in photosynthesis (PS) gene expression and its interaction(s) with other regulators of PS gene expression are of considerable importance to our understanding of the regulatory circuitry governing spectral complex formation. Using a CcoP and FnrL double mutant strain, we obtained results which suggested that FnrL is not involved in the transduction of the inhibitory signal, by which PS gene expression is "silenced," emanating from the cbb(3) oxidase encoded by the ccoNOQP operon under aerobic conditions. The dominant effect of the ccoP mutation in the FnrL mutant strain with respect to spectral complex formation under aerobic conditions and restoration of a PS-positive phenotype suggested that inactivation of the cbb(3) oxidase to some extent bypasses the requirement for FnrL in the formation of spectral complexes. Additional analyses revealed that anaerobic induction of the bchE, hemN, and hemZ genes, which are involved in the tetrapyrrole biosynthetic pathways, requires FnrL. Thus, FnrL appears to be involved at multiple loci involved in the regulation of PS gene expression. Additionally, bchE was also shown to be regulated by the PrrBA two-component system, in conjunction with hemN and hemZ. These and other results to be discussed permit us to more accurately describe the role of FnrL as well as the interactions between the FnrL, PrrBA, and other regulatory circuits in the regulation of PS gene expression.


Low Prevalence of Screen-Detected Colorectal Cancer in an Average-Risk Population: The New Normal.
Prior studies have reported the prevalence of colorectal cancer (CRC) in average-risk screening population ages 50-75 to be 0.7%-1.0%.1,2 However, no estimates from studies enrolling individuals undergoing screening colonoscopy have been reported. The experience of ongoing studies enrolling average-risk individuals is that the prevalence rates are substantially lower. A 2020 study from a community-based cohort undergoing CRC screening with fecal immunochemical testing followed by diagnostic colonoscopy reported a CRC prevalence rate of 1.46 per 1000, or 0.15%.3 The aim of our study is to report the screen-detected prevalence of CRC and advanced neoplasia in average-risk asymptomatic individuals from selected academic and community medical centers in the United States, Canada, and Germany and describe associated risk factors.


Polarized signal classification by complex and quaternionic multi-layer perceptrons.
For polarized signals, which arise in many application fields, a statistical framework in terms of quaternionic random processes is proposed. Based on it, the ability of real-, complex- and quaternionic-valued multi-layer perceptrons (MLPs) of performing classification tasks for such signals is evaluated. For the multi-dimensional neural networks the relevance of class label representations is discussed. For signal to noise separation it is shown that the quaternionic MLP yields an optimal solution. Results on the classification of two different polarized signals are also reported.


The Adenylyl Cyclase Activator Forskolin Increases Influenza Virus Propagation in MDCK Cells by Regulating ERK1/2 Activity.
Vaccination is the most effective method for preventing the spread of the influenza virus. Cell-based influenza vaccines have been developed to overcome the disadvantages of egg-based vaccines and their production efficiency has been previously discussed. In this study, we investigated whether treatment with forskolin (FSK), an adenylyl cyclase activator, affected the output of a cell-based influenza vaccine. We found that FSK increased the propagation of three influenza virus subtypes (A/H1N1/California/4/09, A/H3N2/Mississippi/1/85, and B/Shandong/7/97) in Madin-Darby canine kidney (MDCK) cells. Interestingly, FSK suppressed the growth of MDCK cells. This effect could be a result of protein kinase A (PKA)-Src axis activation, which downregulates extracellular signal-regulated kinase (ERK)1/2 activity and delays cell cycle progression from G1 to S. This delay in cell growth might benefit the binding and entry of the influenza virus in the early stages of viral replication. In contrast, FSK dramatically upregulated ERK1/2 activity via the cAMP-PKA-Raf-1 axis at a late stage of viral replication. Thus, increased ERK1/2 activity might contribute to increased viral ribonucleoprotein export and influenza virus propagation. The increase in viral titer induced by FSK could be explained by the action of cAMP in assisting the entry and binding of the influenza virus. Therefore, FSK addition to cell culture systems could help increase the production efficiency of cell-based vaccines against the influenza virus.


Monitoring and control of amygdala neurofeedback involves distributed information processing in the human brain.
Brain-computer interfaces provide conscious access to neural activity by means of brain-derived feedback ("neurofeedback"). An individual's abilities to monitor and control feedback are two necessary processes for effective neurofeedback therapy, yet their underlying functional neuroanatomy is still being debated. In this study, healthy subjects received visual feedback from their amygdala response to negative pictures. Activation and functional connectivity were analyzed to disentangle the role of brain regions in different processes. Feedback monitoring was mapped to the thalamus, ventromedial prefrontal cortex (vmPFC), ventral striatum (VS), and rostral PFC. The VS responded to feedback corresponding to instructions while rPFC activity differentiated between conditions and predicted amygdala regulation. Control involved the lateral PFC, anterior cingulate, and insula. Monitoring and control activity overlapped in the VS and thalamus. Extending current neural models of neurofeedback, this study introduces monitoring and control of feedback as anatomically dissociated processes, and suggests their important role in voluntary neuromodulation.


Selective cell propagation via micropatterning of a thermally-activated hydrogel.
The ability to selectively propagate specific cells is fundamentally important to the development of clonal cell populations. Current methods rely on techniques such as limiting dilution, colony picking, and flow cytometry to transfer single cells into single wells, resulting in workflows that are low-throughput, slowed by propagation kinetics, and susceptible to contamination. Here, we developed a method, called selective laser gelation (SLG), to micropattern hydrogels in cell culture media in order to encapsulate specific cells to selectively arrest their growth. This process relies on the inverse gelation of methylcellulose, which forms a hydrogel when heated rather than cooled. Local heating using an infrared laser enables hydrogel micropatterning, while phase transition hysteresis retains the hydrogel after laser excitation. As a demonstration, we used this approach to selectively propagate transgenic CHO cells with increased antibody productivity. More generally, hydrogel micropatterning provides a simple and non-contact method for selective propagation of cells based on features identified by imaging.


Quantum Heat Engine Using Electromagnetically Induced Transparency.
We report an experiment demonstrating the generation of directional thermal radiation with a spectral brightness that is about 9 times greater than that of the ambient pumping reservoir. The experiment is based on the recent proposal for a nontraditional quantum heat engine and uses cold Rb atoms, electromagnetically induced transparency, and photon correlation spectroscopy [Phys. Rev. A 94, 053859 (2016)PLRAAN2469-992610.1103/PhysRevA.94.053859].


Motility of select ovarian cancer cell lines: effect of extra-cellular matrix proteins and the involvement of PAK2.
The interaction between tumor cells and extracellular matrix (ECM) proteins influences cell migration and the invasive behavior of cancer cells. In this study, we provide experimental evidence that collagen I and fibronectin affect ovarian cancer cell migration. In vitro wound healing assays and transwell migration assays were used to measure both total wound healing and directionality of individually migrating OV2008 and C13 ovarian cancer cells on glass, collagen I and fibronectin. Involvement of p21‑activated kinase 2 (Pak2) in the motility of these cell lines was investigated using a chemical inhibitor as well as siRNA transfection. Culturing ovarian cancer cells on collagen type I (COLL) increased the migratory ability of OV2008 and C13 cells by increasing the directional migration of cells. In contrast, fibronectin (FN) decreased the migratory ability of OV2008 cells by reducing their ability to migrate directionally. When both cell lines are cultured on COLL and treated with increasing concentrations of a PAK inhibitor (IPA-3), there is a dose-dependent response such that there is a decrease in migration with an increase in inhibitor concentration. Further experiments utilizing PAK2 knockdown via siRNA transfection demonstrated significantly reduced migration of OV2008 cells on COLL as compared to those receiving control siRNA. In conclusion, our results indicate that FN and COLL affect the motility of the selected ovarian cancer cells lines and the effect of COLL is likely mediated, at least in part, by PAK2. A better understanding of the molecular events that contribute to tumor invasion and metastasis is crucial for developing novel treatment strategies to improve the long-term survival of women with ovarian cancer. As PAK2 is involved in motility, it should be further explored as a pro-metastatic gene in ovarian cancer.


Cow's milk neutralizes the cytotoxicity of acrolein, a putative carcinogen in cigarette smoke.
Cigarette smoke is a strong and independent risk factor for esophageal cancer, while the consumption of cow's milk has been proposed as a protective factor. The mechanistic role of milk in preventing cancer, however, has not been clarified. We focused our study on acrolein, an abundant unsaturated aldehyde present in cigarette smoke. Acrolein is a highly toxic compound and a putative carcinogen. Using a cell culture system, we found that (1) acrolein caused necrosis in Ramos Burkitt's lymphoma cells, (2) the necrosis was inhibited by preincubation of acrolein with milk, and (3) acrolein formed adducts with milk proteins. These results indicated the protective effects of cow's milk against acrolein-induced cytotoxicity via protein-acrolein adduct formation.


Roundtable on Preseason Heat Safety in Secondary School Athletics: Environmental Monitoring During Activities in the Heat.
To develop best-practice recommendations using thermal indices to determine work-to-rest ratios and facilitate further implementation of environmental monitoring for heat safety in secondary school athletics in the United States. A narrative review of the current literature on environmental monitoring for heat safety during athletics was conducted by content experts. A list of action-oriented recommendations was established from the narrative review and further refined using the Delphi method. Assessment of wet bulb globe temperature at the site of activity and throughout the duration of the event is recommended to assist clinicians and administrators in making appropriate decisions regarding the duration and frequency of activity and rest periods. Activity-modification guidelines should be predetermined and approved by stakeholders and should outline specific actions to be followed, such as the work-to-rest ratio, frequency and timing of hydration breaks, and adjustment of total exercise duration, equipment, and clothing. Furthermore, integration of exertional heat illness injury data with environmental condition characteristics is critical for the development of evidence-based heat safety guidelines for secondary school athletics. Athletic trainers play an essential role in conducting prospective injury data collection, recording onsite wet bulb globe temperature levels, and implementing recommendations to protect the health and safety of athletes.


Effectiveness of Different Cervical Range of Motion Measurement Techniques for Home-Use to Prevent Cervical Spondylosis.
Cervical spondylosis is a non-specific degenerative of cervical spine which results in spinal canal and nerve root foramen stenosis. The stenosis of the canals results in injury of spinal cord and nerve root. The nerve root compression causes a various symptom, such as referred pain and numbness in neck and upper extremities. Motion sensors allow for the tracking and observation of cervical movement activities with the purpose of preventing cervical spondylosis. In the proposed study, Inertia Measurement Unit (IMU) sensors and comparative 2- Dimensional Motion Capture (2D-MC) system were considered to determine the effective of cervical range of motion in various environments. The results indicated that both methods provided strong correlations of craniovertebral angles, with the IMU sensors showing a higher correlation coefficient than the 2D-MC system. Therefore, the craniovertebral angles from IMU sensors were utilized to identify the safety and warning zones of neck movements.Clinical Relevance- The degenerative of the cervical spine results in different degrees of severity in cervical spondylosis. To prevent further deterioration, it is recommended to adopt lifestyle changes, especially neck movement changes, that reduce the spinal cord or nerve root compression. An innovation that can detect harmful neck movements in real-time can provide feedback to users on whether they are moving their head into dangerous angles. By training regularly with this innovation, individuals can delay the onset and severity of cervical spondylosis symptoms and make adjustments to their lifestyles to prevent recurrence of the condition in the future.


Selective Interleukin-23 p19 Inhibition: Another Game Changer in Psoriasis? Focus on Risankizumab.
The history of psoriasis treatment has been marked by several milestones. Corticosteroids, cyclosporine, tumor necrosis factor alpha (TNF-α) inhibitors and, more recently, interleukin (IL)-17A inhibitors have revolutionized the treatment of psoriasis, each in its own way and time. The IL-23/IL-17 axis is currently considered to be crucial in the pathogenesis of psoriasis and selective IL-23p19 inhibition may bring several advantages with respect to IL-12/23p40 inhibition, or distal blockade of IL-17A or its receptor. In fact, IL-12 axis inhibition does not appear to be essential in psoriasis and IL-12 inhibition may even have a negative effect in the treatment of psoriasis and have potential risks in tumor immune surveillance and in host defense against intracellular pathogens. On the other hand, contrary to IL-17 inhibition, IL-23p19 blockade does not increase the risk of candida infection, nor is it associated with inflammatory bowel disease worsening. Several IL-23p19 inhibitors are currently being developed for the treatment of psoriasis, such as tildrakizumab, guselkumab, and risankizumab. Although clinical data on risankizumab is still scarce, it has shown characteristics that signify a major advance in the treatment of this disease, offering comparable or higher efficacy than IL-17 inhibitors, without the safety concerns of this therapeutic class, combined with the excellent dosing regimen of ustekinumab. Currently, only phase II trial data is available; thus, the results of the large phase III trials will be essential to establish the efficacy and safety profile of risankizumab and its value in the biological armamentarium for the treatment of psoriatic patients.


Pharmacological, hemodynamic and biochemical mechanisms involved in the blood pressure lowering effects of pergolide, in normotensive and hypertensive dogs.
In pentobarbital-anesthetized normotensive dogs, clonidine (20.0 micrograms/kg i.v.), in contrast to pergolide (30.0 micrograms/kg i.v.), reduced significantly both aortic blood pressure and plasma concentration of norepinephrine. However, in dogs that had been made hypertensive by sectioning the vagi and carotid sinus nerves, pergolide, like clonidine, lowered the blood pressure and plasma concentrations of epinephrine and norepinephrine that were enhanced markedly by deafferentation. Furthermore, in this preparation pergolide decreased the calculated resistance in vascular regions supplied by the upper abdominal aorta and the innervated femoral and renal arteries, but it increased vascular resistance in the denervated hind leg. Pergolide (1.0 microgram/kg) injected intracisternally (i.c.m.) induced a fall in blood pressure of comparable magnitude to that produced by a 30 times higher i.v. dose. Intravenously and i.c.m. administered pergolide lowered blood pressure by acting at distinct anatomical sites inasmuch as i.v. sulpiride blocked the effects of i.v. but not i.c.m. pergolide. The combination of sulpiride plus yohimbine injected i.c.m. was necessary to abolish the decrease in blood pressure evoked by i.c.m. pergolide. In atropinized spinal dogs, i.v. pergolide inhibited the vasoconstriction elicited by electrical stimulation of the lumbar sympathetic chain, an effect which was antagonized by sulpiride. Similarly, pergolide (30.0 micrograms/kg i.v.) like clonidine, reduced the heart rate and coronary venous plasma norepinephrine concentration raised by sustained electrical stimulation of the cardioaccelerator nerve. Sulpiride, but not phentolamine, antagonized this pergolide-induced inhibition of sympathetic nerve function. In chlorisondamine-pretreated dogs, pergolide produced a transient pressor response due to stimulation of postsynaptic vascular alpha-2 adrenoceptors. In conclusion, the failure of i.v. pergolide to decrease aortic blood pressure in pentobarbital-anesthetized normotensive dogs is presumably due to the inability of pergolide to produce a significant inhibition of the vascular sympathetic tone in this preparation. However, in neurogenic hypertensive dogs which are characterized by an elevated level of sympathetic drive, i.v. pergolide reduced blood pressure and aortic plasma norepinephrine concentration. These effects of pergolide are compatible with a DA-2 dopamine receptor stimulation on peripheral sympathetic nerve fibers. In contrast, the antihypertensive effects of i.c.m. pergolide would appear to be mediated by both alpha-2 adrenoceptors and DA-2 dopamine receptors located within the central nervous system.


L-Aminoacid Oxidase from Bothrops leucurus Venom Induces Nephrotoxicity via Apoptosis and Necrosis.
Acute renal failure is a common complication caused by Bothrops viper envenomation. In this study, the nefrotoxicity of a main component of B. leucurus venom called L-aminoacid oxidase (LAAO-Bl) was evaluated by using tubular epithelial cell lines MDCK and HK-2 and perfused kidney from rats. LAAO-Bl exhibited cytotoxicity, inducing apoptosis and necrosis in MDCK and HK-2 cell lines in a concentration-dependent manner. MDCK apoptosis induction was accompanied by Ca2+ release from the endoplasmic reticulum, reactive oxygen species (ROS) generation and mitochondrial dysfunction with enhanced expression of Bax protein levels. LAAO-Bl induced caspase-3 and caspase-7 activation in both cell lines. LAAO-Bl (10 μg/mL) exerts significant effects on the isolated kidney perfusion increasing perfusion pressure and urinary flow and decreasing the glomerular filtration rate and sodium, potassium and chloride tubular transport. Taken together our results suggest that LAAO-Bl is responsible for the nephrotoxicity observed in the envenomation by snakebites. Moreover, the cytotoxic of LAAO-Bl to renal epithelial cells might be responsible, at least in part, for the nephrotoxicity observed in isolated kidney.


Effects of protein and fat concentration in coproduct-based growing calf diets on adipogenic and lipogenic gene expression, blood metabolites, and carcass composition.
Crossbred calves ( = 30; age = 95 ± 1.7 d; BW = 179 ± 18 kg) were fed 1 of 5 growing diets: 1) corn-based control, 2) low-fat, low-protein coproduct blend, 3) high-fat, low-protein coproduct blend, 4) low-fat, high-protein coproduct blend, and 5) high-fat, high-protein coproduct blend for 112 d (growing phase) followed by a common corn-based finishing diet (additional 112 d; finishing phase). Calves were biopsied at 0, 112, and 224 d for transcriptional analysis via real-time quantitative PCR of 14 genes associated with adipogenesis and lipogenesis within the muscle. Serum was collected at d 0, 112, and 224 and analyzed for leptin, IGF-1, and GH concentration. Treatments were arranged in a 2 × 2 factorial. Data were analyzed using the MIXED procedures of SAS (SAS Inst. Inc., Cary, NC) to ascertain the effects of 2 protein levels, 2 fat levels, time, and any interactions. Increased protein and decreased fat in the growing diet resulted in a carryover effect that increased ( 0.01) gene expression of PPARγ, insulin-induced gene 1, thyroid hormone responsive SPOT14 protein, ATP citrate lyase, adiponectin, diacylglycerol O-acyltransferase homologue 2, fatty acid binding protein 4, fatty acid synthase, phosphoenolpyruvate carboxykinase 1, and stearoyl-CoA desaturase as well as serum leptin concentrations between d 112 and 224. Expression of sterol regulatory element binding transcription factor 1 was increased ( 0.01) at d 112 in steers fed high-protein, high-fat diets compared to those fed high-protein, low-fat diets. A fat × day interaction ( 0.01) occurred for the expression of adiponectin receptor 2 and CCAAT/enhancer binding protein alpha, resulting in a carryover effect wherein low-fat diets fed during the growing phase increased expression of both genes at the end of the finishing phase (d 224). After slaughter, cattle fed the control during the growing phase tended ( 0.09) to have greater marbling scores, whereas other carcass parameters were not different ( ≥ 0.13). These data indicate that feeding differing levels of dietary fat and protein during the growing phase does affect i.m. adipogenesis at the transcriptional level, but differences in gene expression were not sufficient to affect carcass quality among cattle fed coproducts.


Preserved Motility after Neonatal Dopaminergic Lesion Relates to Hyperexcitability of Direct Pathway Medium Spiny Neurons.
In Parkinson's disease patients and rodent models, dopaminergic neuron loss (DAN) results in severe motor disabilities. In contrast, general motility is preserved after early postnatal DAN loss in rodents. Here we used mice of both sexes to show that the preserved motility observed after early DAN loss depends on functional changes taking place in medium spiny neurons (MSN) of the dorsomedial striatum (DMS) that belong to the direct pathway (dMSN). Previous animal model studies showed that adult loss of dopaminergic input depresses dMSN response to cortical input, which likely contributes to Parkinson's disease motor impairments. However, the response of DMS-dMSN to their preferred medial PFC input is preserved after neonatal DAN loss as shown by in vivo studies. Moreover, their response to inputs from adjacent cortical areas is increased, resulting in reduced cortical inputs selectivity. Additional ex vivo studies show that membrane excitability increases in dMSN. Furthermore, chemogenetic inhibition of DMS-dMSN has a more marked inhibitory effect on general motility in lesioned mice than in their control littermates, indicating that expression of normal levels of locomotion and general motility depend on dMSN activity after early DAN loss. Contrastingly, DMS-dMSN inhibition did not ameliorate a characteristic phenotype of the DAN-lesioned animals in a marble burying task demanding higher behavioral control. Thus, increased dMSN excitability likely promoting changes in corticostriatal functional connectivity may contribute to the distinctive behavioral phenotype emerging after developmental DAN loss, with implications for our understanding of the age-dependent effects of forebrain dopamine depletion and neurodevelopment disorders.SIGNIFICANCE STATEMENT The loss of striatal dopamine in the adult brain leads to life-threatening motor impairments. However, general motility remains largely unaffected after its early postnatal loss. Here, we show that the high responsiveness to cortical input of striatal neurons belonging to the direct basal ganglia pathway, crucial for proper motor functioning, is preserved after early dopamine neuron loss, in parallel with an increase in these cells' membrane excitability. Chemogenetic inhibition studies show that the preserved motility depends on this direct pathway hyperexcitability/hyperconnectivity, while other phenotypes characteristic of this condition remained unaltered despite the dMSN inhibition. This insight has implications for our understanding of the mechanism underlying the behavioral impairments observed in neuropsychiatric conditions linked to early dopaminergic hypofunction.


Genetic substructuring as a result of barriers to gene flow in urban Rana temporaria (common frog) populations: implications for biodiversity conservation.
The ability to maintain small populations in quasi-natural settings is an issue of considerable importance in biodiversity conservation. The genetic structure of urban common frog (Rana temporaria) populations was determined by allozyme electrophoresis and used to evaluate the effects of restricted intersite migration. Despite the lack of any absolute barrier to movement between ponds, substantial genetic differentiation was found between sites separated by an average of only 2.3 km. Genetic distances between these town ponds correlated positively with geographical distances and were almost twice as great as those found between rural sites separated by an average of 41 km. Measures of genetic diversity and fitness were always lowest in the town, where the degree of subpopulation differentiation (FST = 0.388) was high. Population decline was not evident in the town, but molecular and fitness data indicated the presence of genetic drift and inbreeding depression. The long-term survival of artificially restricted populations, particularly of relatively sedentary species, may require molecular monitoring, if genetic diversity is not to be lost by chance when facets of the species niche prove to be poorly understood.


Ly49Q positively regulates type I IFN production by plasmacytoid dendritic cells in an immunoreceptor tyrosine-based inhibitory motif-dependent manner.
Plasmacytoid dendritic cells (pDC) are the major producers of type I IFN during the initial immune response to viral infection. Ly49Q, a C-type lectin-like receptor specific for MHC-I, possesses a cytoplasmic ITIM and is highly expressed on murine pDC. Using Ly49Q-deficient mice, we show that, regardless of strain background, this receptor is required for maximum IFN-α production by pDC. Furthermore, Ly49Q expression on pDC, but not myeloid dendritic cells, is necessary for optimal IL-12 secretion, MHC-II expression, activation of CD4(+) T cell proliferation, and nuclear translocation of the master IFN-α regulator IFN regulatory factor 7 in response to TLR9 agonists. In contrast, the absence of Ly49Q did not affect plasmacytoid dendritic cell-triggering receptor expressed on myeloid cells expression or pDC viability. Genetic complementation revealed that IFN-α production by pDC is dependent on an intact tyrosine residue in the Ly49Q cytoplasmic ITIM. However, pharmacological inhibitors and phosphatase-deficient mice indicate that Src homology 2 domain-containing phosphatase 1 (SHP)-1, SHP-2, and SHIP phosphatase activity is dispensable for this function. Finally, we observed that Ly49Q itself is downregulated on pDC in response to CpG exposure in an ITIM-independent manner. In conclusion, Ly49Q enhances TLR9-mediated signaling events, leading to IFN regulatory factor 7 nuclear translocation and expression of IFN-I genes in an ITIM-dependent manner that can proceed without the involvement of SHP-1, SHP-2, and SHIP.


The Effect of Bleaching on the Basic Colour and Discoloration Susceptibility of Dental Composites.
To test the influence of a bleaching procedure using 16% carbamide peroxide (CP) on the colour of composite materials and their consecutive subjection to discolouration in beverages. Nanocomposite Z550 (3M ESPE) and the microhybrid Z250 (3M ESPE) composite materials were selected for the research. 16% CP was applied to composite plates (15 samples each material) for seven hours a day for 14 days simulating at-home nightguard vital bleaching. The test samples were then divided randomly into three sub-groups and submerged in instant coffee, green tea and Coca Cola for 30 days. The control group (N=5) samples were kept in deionized water. Determination of the L*a*b* dimensions of colour of the polymerized discs was performed by calibrated spectrophotometer 24 hours later, after the bleaching procedure, and on days 7, 15 and 30 after immersion in the beverages. After application of 16% CP, there were perceptible changes in the colour of both test materials, which did not exceed the boundaries of acceptability (ΔE<3.48). Coca-Cola did not cause discolouration of the tested composites, but coffee and tea changed their colour above the level of perceptibility already seven days after immersion. A statistically significant difference in the change in colour was established between the materials when they were immersed in coffee (P<0.05). 16% CP does not affect the basic colour of the composites. Immersion in a beverage led to an unacceptable change in the colour of both test materials in coffee and tea, primarily towards black. The greatest discolouration after bleaching was shown by Z550 when immersed in coffee.


[Atrial fibrillation after mitral valve replacement].
Atrial fibrillation at patients with mitral valve disease is a common complication, both before and after surgery. The analysis of factors associated with the risk of atrial fibrillation in patients with mitral valvular disease was perfomed. It has been established that the conduct of surgery in patients with mitral valve contributes not only to improve the clinical course of the disease, but also the restoration of sinus rhythm.


[Initial experience with the use of the proton beam at the Institute of Theoretical and Experimental Physics to treat prostatic cancer].
The proton beam of the 157 MeV synchrotron of the Institute of Theoretical and Experimental Physics was modified for irradiation of prostatic cancer patients. Patient's topometric preparation and method of transperineal irradiation were described. A method of radiotherapy with proton beam boost was applied to 17 patients with different clinical stages of disease. After 6-20 months of the follow-up all the patients have been alive without clinical signs of tumor. In most of the patients radiation reactions of the critical organs were regarded as mild ones. Chronic and long-term radiation injuries were not detected in the patients who lived over 1 year.


The Effects of Statin Therapy on Oxidized LDL and Its Antibodies: A Systematic Review and Meta-Analysis.
Elevated serum low-density lipoproteins (LDL), the substrate for the formation of atherogenic oxidized LDLs (oxLDL), are a causal factor for atherosclerotic cardiovascular disease (ASCVD). Statins are well known to decrease LDL particle concentration and reduce ASCVD morbidity and mortality. To perform a meta-analysis of the effects of statins (i.e., type, dose, and duration of treatment) on serum levels of oxLDL and on immunoglobulin M (IgM) and immunoglobulin G (IgG) antibody levels against oxLDL. PubMed, Scopus, Embase, and Web of Science were searched up to February 5th, 2021, for randomized controlled trials (RCT) evaluating the effect of statins on oxLDL and anti-oxLDL antibody levels. Meta-analysis was performed using Comprehensive Meta-Analysis (CMA) V2 software. To evaluate the influence of each study on the overall effect size, a sensitivity analysis was performed using the leave-one-out method. Evaluation of the funnel plot, Begg's rank correlation, and Egger's weighted regression tests was used to assess the presence of publication bias in the meta-analysis. A total of 28 RCTs including 4019 subjects were finally included in the meta-analysis. The results indicated a significant decrease in circulating concentrations of oxLDL after treatment with statins (SMD: -2.150, 95% CI: -2.640, -1.697, p < 0.001). Subgroup analysis found no significant effect of the intensity of statin treatment or statin lipophilicity on the reduction of circulating concentrations of oxLDL. An additional meta-analysis of 3 trials showed that statins did not change the serum levels of IgM and IgG antibodies to oxLDL. Statin therapy decreases serum oxLDL concentrations but does not affect circulating levels of anti-oxLDL antibodies.


Mutation Survey of Candidate Genes and Genotype-Phenotype Analysis in 20 Southeastern Chinese Patients with Axenfeld-Rieger Syndrome.
Purpose/aim of the study: To conduct a survey spectrum of the PITX2, FOXC1, and PRDM5 genes to reveal genotype-phenotype correlations in a cohort of Southeastern Chinese patients with Axenfeld-Rieger syndrome (ARS). A total of 20 probands with ARS were recruited in Southeast China. All patients underwent full ocular and systemic examinations. Sanger sequencing was used to analyze all coding regions of and regions adjacent to PITX2, FOXC1, and PRDM5 and 13 upstream regulatory elements of PITX2. Multiplex ligation-dependent probe amplification was performed to detect gross insertions and deletions in PITX2 and FOXC1. Quantitative polymerase chain reaction was used to detect copy number variations in regulatory elements of PITX2. A bioinformatics analysis was conducted to evaluate the pathogenicity of variants. Eleven mutations, including eight novel mutations, were identified in PITX2. Seven of the mutations were truncations. A genotype-phenotype correlation analysis showed that 81.8% (9/11) of patients with mutations in PITX2 developed glaucoma before reaching 10 years old. The proportion of patients without detected mutations was only 33.3% (3/9, P = 0.0399). In patient G1399, ultrasound biomicroscopy revealed that the left eye exhibited a phenotype similar to aniridia with complete angle closure and a remaining stub of iris tissue. This is the first genetic study of a cohort of Southeastern Chinese patients with ARS. Eight novel mutations were detected, expanding the mutation spectrum of PITX2. PITX2 may be a major candidate gene for ARS in Southeastern Chinese patients. Truncations may be the primary mutation type in PITX2. Glaucoma onset may be earlier in patients with mutations in PITX2 than in those without mutations in PITX2 and FOXC1. A block of the anterior chamber angle by the end of the iris might represent the main factor influencing the development of glaucoma in ARS patients with an asymmetric aniridia phenotype.


Expression and purification of the cystic fibrosis transmembrane conductance regulator protein in Saccharomyces cerevisiae.
The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel, that when mutated, can give rise to cystic fibrosis in humans.There is therefore considerable interest in this protein, but efforts to study its structure and activity have been hampered by the difficulty of expressing and purifying sufficient amounts of the protein(1-3). Like many 'difficult' eukaryotic membrane proteins, expression in a fast-growing organism is desirable, but challenging, and in the yeast S. cerevisiae, so far low amounts were obtained and rapid degradation of the recombinant protein was observed (4-9). Proteins involved in the processing of recombinant CFTR in yeast have been described(6-9) .In this report we describe a methodology for expression of CFTR in yeast and its purification in significant amounts. The protocol describes how the earlier proteolysis problems can be overcome and how expression levels of CFTR can be greatly improved by modifying the cell growth conditions and by controlling the induction conditions, in particular the time period prior to cell harvesting. The reagants associated with this protocol (murine CFTR-expressing yeast cells or yeast plasmids) will be distributed via the US Cystic Fibrosis Foundation, which has sponsored the research. An article describing the design and synthesis of the CFTR construct employed in this report will be published separately (Urbatsch, I.; Thibodeau, P. et al., unpublished). In this article we will explain our method beginning with the transformation of the yeast cells with the CFTR construct - containing yeast plasmid (Fig. 1). The construct has a green fluorescent protein (GFP) sequence fused to CFTR at its C-terminus and follows the system developed by Drew et al. (2008)(10). The GFP allows the expression and purification of CFTR to be followed relatively easily. The JoVE visualized protocol finishes after the preparation of microsomes from the yeast cells, although we include some suggestions for purification of the protein from the microsomes. Readers may wish to add their own modifications to the microsome purification procedure, dependent on the final experiments to be carried out with the protein and the local equipment available to them. The yeast-expressed CFTR protein can be partially purified using metal ion affinity chromatography, using an intrinsic polyhistidine purification tag. Subsequent size-exclusion chromatography yields a protein that appears to be >90% pure, as judged by SDS-PAGE and Coomassie-staining of the gel.


Functional MYB transcription factor gene HtMYB2 is associated with anthocyanin biosynthesis in Helianthus tuberosus L.
Tuber color is an important trait for Helianthus tuberosus L. (Jerusalem artichoke). Usually, purple tubers with high anthocyanin content are more nutritious than white tuber. But, the molecular mechanism underlying it is unknown. In the current study, high-throughput RNA-sequencing was used to compare the transcriptomes between plants with tubers with red or white epidermis. Compared with the white-skinned tubers of cultivar QY3, anthocyanin biosynthesis structural genes had greater expression in the red-skinned tubers of cultivar QY1, indicating that the anthocyanin biosynthesis pathway was activated in 'QY1'; quantitative PCR confirmed this difference in expression. HtMYB2 (Unigene44371_All) was the only MYB transcription factor, homologous to the MYB transcription factor regulating anthocyanin biosynthesis, expressed in the red tuber epidermis of 'QY1'. The anthocyanin concentration in the root, stem, leaf, flower, and tuber epidermis of 'QY1' was higher than in 'QY3', especially tuber epidermis. Correspondingly, HtMYB2 had greater expression in these tissues of 'QY1' than in 'QY3'. The expression of HtMYB2 was associated with anthocyanin accumulation in the different tissues. Overexpression of HtMYB2 activated the anthocyanin biosynthesis pathway, accumulating the pigment in leaves of transgenic tobacco, supporting the model that HtMYB2 regulated anthocyanin biosynthesis. Further experiments found that HtMYB2 had the same coding sequence and genomic sequence in 'QY1' and 'QY3', but that there were several single nucleotide polymorphisms and one insertion-deletion (indel) mutation of 21 nucleotides in the promoter region between the two alleles. The deletion of three nucleotides "AAA" made the promoter of 'QY1' predicted to contain one more possible promoter region. A specific primer, based on the indel, could differentiate between cultivars with red or white tuber epidermis. The genetic variation in HtMYB2 was associated with the tuber skin color in a natural population. RNA-seq can successfully isolate the candidate gene (HTMYB2) controlling anthocyanin biosynthesis in purple epidermis of Jerusalem artichoke tuber. HTMYB2 can regulate anthocyanin biosynthesis in plants and is closely related to the formation of purple phenotype in tubers. This study should be useful in understanding the genetic mechanism underlying different tuber skin colors and in breeding new H. tuberosus cultivars with different tuber skin colors.


Severity of Peri-ictal Respiratory Dysfunction With Epilepsy Duration and Patient Age at Epilepsy Onset.
Respiratory dysfunction preceding death is fundamental in sudden unexpected death in epilepsy (SUDEP) pathophysiology. Hypoxia occurs with one-third of seizures. In temporal lobe epilepsy, there is volume loss in brainstem regions involved in autonomic control and increasing neuropathological changes with duration of epilepsy suggesting increasingly impaired regulation of ventilation. In animal models, recurrent hypoxic episodes induce long-term facilitation (LTF) of ventilatory function, however, LTF is less robust in older animals. LTF of ventilation may, to some degree, ameliorate the deleterious effects of progressive brainstem atrophy. We investigated the possibility that the duration of epilepsy, or age at epilepsy onset, may impact the severity of seizure-associated respiratory dysfunction. Patients with focal epilepsy undergoing video-EEG telemetry in the epilepsy monitoring unit (EMU) were studied. We found a significant relationship between age at epilepsy onset and duration of peri-ictal oxygen desaturation for focal seizures not progressing to bilateral tonic-clonic seizures, with longer duration of peri-ictal oxygen desaturation in patients with epilepsy onset at an older age but no significant relationships between duration of epilepsy or age at EMU admission and ventilatory dysfunction. Our findings suggest an intriguing possibility that LTF of ventilation may be protective when epilepsy starts at a younger age.


Spatial properties of twin-beam correlations at low- to high-intensity transition.
It is shown that spatial correlation functions measured for correlated photon pairs at the single-photon level correspond to speckle patterns visible at high intensities. This correspondence is observed for the first time in one experimental setup by using different acquisition modes of an intensified CCD camera in low and high intensity regimes. The behavior of intensity auto- and cross-correlation functions in dependence on pump-beam parameters including power and transverse profile is investigated.


Functional characteristics and tissue distribution pattern of organic cation transporter 2 (OCTN2), an organic cation/carnitine transporter.
We have demonstrated in the present study that novel organic cation transporter (OCTN) 2 is a transporter for organic cations as well as carnitine. OCTN2 transports organic cations without involving Na(+), but it transports carnitine only in the presence of Na(+). The ability to transport organic cations and carnitine is demonstrable with human, rat, and mouse OCTN2s. Na(+) does not influence the affinity of OCTN2 for organic cations, but it increases the affinity severalfold for carnitine. The short-chain acyl esters of carnitine are also transported by OCTN2. Two mutations, M352R and P478L, in human OCTN2 are associated with loss of transport function, but the protein expression of these mutants is comparable to that of the wild-type human OCTN2. In situ hybridization in the rat shows that OCTN2 is expressed in the proximal and distal tubules and in the glomeruli in the kidney, in the myocardium, valves, and arterioles in the heart, in the labyrinthine layer of the placenta, and in the cortex, hippocampus, and cerebellum in the brain. This is the first report that OCTN2 is a Na(+)-independent organic cation transporter as well as a Na(+)-dependent carnitine transporter and that OCTN2 is expressed not only in the heart, kidney, and placenta but also in the brain.


MicroRNAs.
MicroRNAs (miRNAs) are small, noncoding RNAs with regulatory functions, which play an important role in many human diseases, including cancer. An emerging number of studies show that miRNAs can act either as oncogenes or as tumor suppressor genes or sometimes as both. Germline, somatic mutations and polymorphisms can contribute to cancer predisposition. miRNA expression levels have diagnostic and prognostic implications, and their roles as anticancer therapeutic agents is promising and currently under investigation.


Automated immunohistochemical assessment ability to evaluate estrogen and progesterone receptor status compared with quantitative reverse transcription-polymerase chain reaction in breast carcinoma patients.
This study aimed to investigate the capability of an automated immunohistochemical (IHC) evaluation of hormonal receptor status in breast cancer patients compared to a well-validated quantitative reverse transcription-polymerase chain reaction (RT-qPCR) method. This study included 93 invasive breast carcinoma cases that had both standard IHC assay and Oncotype Dx assay results. The same paraffin blocks on which Oncotype Dx assay had been performed were selected. Estrogen receptor (ER) and progesterone receptor (PR) receptor status were evaluated through IHC stains using SP1 monoclonal antibody for ER, and 1E2 monoclonal antibody for PR. All ER and PR immunostained slides were scanned, and invasive tumor areas were marked. Using the QuantCenter image analyzer provided by 3DHISTECH, IHC staining of hormone receptors was measured and converted to histochemical scores (H scores). Pearson correlation coefficients were calculated between Oncotype Dx hormone receptor scores and H scores, and between Oncotype Dx scores and Allred scores. H scores measured by an automated imaging system showed high concordance with RT-qPCR scores. ER concordance was 98.9% (92/93), and PR concordance was 91.4% (85/93). The correlation magnitude between automated H scores and RT-qPCR scores was high and comparable to those of Allred scores (for ER, 0.51 vs. 0.37 [p=.121], for PR, 0.70 vs. 0.72 [p=.39]). Automated H scores showed a high concordance with quantitative mRNA expression levels measured by RT-qPCR.


Childhood sarcoidosis: A rare but fascinating disorder.
Childhood sarcoidosis is a rare multisystemic granulomatous disorder of unknown etiology. In the pediatric series reported from the southeastern United States, sarcoidosis had a higher incidence among African Americans. Most reported childhood cases have occurred in patients aged 13-15 years. Macrophages bearing an increased expression of major histocompatibility class (MHC) II molecules most likely initiate the inflammatory response of sarcoidosis by presenting an unidentified antigen to CD4+ Th (helper-inducer) lymphocytes. A persistent, poorly degradable antigen driven cell-mediated immune response leads to a cytokine cascade, to granuloma formation, and eventually to fibrosis. Frequently observed immunologic features include depression of cutaneous delayed-type hypersensitivity and a heightened helper T cell type 1 (Th1) immune response at sites of disease. Circulating immune complexes, along with signs of B cell hyperactivity, may also be found. The clinical presentation can vary greatly depending upon the organs involved and age of the patient. Two distinct forms of sarcoidosis exist in children. Older children usually present with a multisystem disease similar to the adult manifestations, with frequent hilar lymphadenopathy and pulmonary infiltrations. Early-onset sarcoidosis is a unique form of the disease characterized by the triad of rash, uveitis, and arthritis in children presenting before four years of age. The diagnosis of sarcoidosis is confirmed by demonstrating a typical noncaseating granuloma on a biopsy specimen. Other granulmatous diseases should be reasonably excluded. The current therapy of choice for sarcoidosis in children with multisystem involvement is oral corticosteroids. Methotrexate given orally in low doses has been effective, safe and steroid sparing in some patients. Alternative immunosuppressive agents, such as azathioprine, cyclophosphamide, chlorambucil, and cyclosporine, have been tried in adult cases of sarcoidosis with questionable efficacy. The high toxicity profile of these agents, including an increased risk of lymphoproliferative disorders and carcinomas, has limited their use to patients with severe disease refractory to other agents. Successful steroid sparing treatment with mycophenolate mofetil was described in an adolescent with renal-limited sarcoidosis complicated by renal failure. Novel treatment strategies for sarcoidosis have been developed including the use of TNF-alpha inhibitors, such as infliximab. The long-term course and prognosis is not well established in childhood sarcoidosis, but it appears to be poorer in early-onset disease.


Implications of Increased Weight Status for the Occurrence of Fall-Induced Intracranial Hemorrhage in Children Aged 4 Years or Younger.
The aim of this study was to investigate the association between increased weight status (IWS), a weight for age/sex at greater than the 95th percentile, and fall-induced intracranial hemorrhage (ICH) in children aged 4 years or younger. In 7072 children aged 4 years or younger with head injury who visited a tertiary care hospital emergency department in Korea from 2013 through 2015, the presence of fall-induced ICH was reviewed. The association between IWS and ICH was investigated by multivariable logistic regression. We retrospectively validated the Pediatric Emergency Care Applied Research Network rule alone and in combination with IWS for predicting ICH. Of 7072 children, 547 (7.7%) underwent computed tomography, of whom 451 (6.4%) were enrolled. Of these, 41 (9.1%; estimated event rate, 0.6%) had ICHs, and 26 (5.8%) had IWS. Increased weight status was more common in the children with ICH (P = 0.023). The association between IWS and ICH remained significant after adjustment (odds ratio, 5.24; 95% confidence interval [CI], 1.49-18.46; P = 0.010). The validation of the rule in combination with IWS showed no significant increases in a sensitivity (92.7% [95% CI, 80.1%-98.5%] to 95.7% [95% CI, 83.5%-99.4%]) and negative predictive value (98.2% [95% CI, 94.7%-99.4%] to 98.8% [95% CI, 95.4%-99.8%]). Increased weight status is associated with fall-induced ICH in children aged 4 years or younger. Information on weight status could be potentially helpful in predicting ICH in young children with fall-induced head injury.


Unsuspected strongyloidiasis in hospitalised elderly patients with and without eosinophilia.
The prevalence and associated factors of chronic uncomplicated strongyloidiasis were estimated among 200 consecutive elderly patients (aged >or= 60 years) admitted to a general hospital in northern Italy. One-hundred patients had a peripheral eosinophil concentration >or= 500 cells/microL (group A), and 100 were age- and gender-matched controls (group B). Measurements included serum IgG anti-Strongyloides antibody titre by an indirect immunofluorescence assay, combined with faecal culture for Strongyloides stercoralis. Anti-Strongyloides antibodies were detected in 28 patients (at high titre in 11 patients). Seropositivity was significantly more common among group A than among group B patients (OR 4.85). Strong seropositivity for anti-Strongyloides antibodies was associated with farm work (p < 0.001), but not with other patient characteristics or with signs and symptoms of strongyloidiasis. In conclusion, strongyloidiasis was relatively common among elderly in-patients; eosinophilia and a history of farm work were the most useful indications for this diagnosis.


Evaluation of the (1,3)-β-D-glucan assay for the diagnosis of neonatal invasive yeast infections.
Most newborns in the neonatal intensive care unit (NICU) are premature and at risk of invasive fungal infections (IFIs). Invasive yeast infections (IYIs) are the most common fungal infections in this population. These infections are difficult to diagnose because symptoms are nonspecific, and the sensitivity of blood cultures is low. The serum (1,3)-β-D-glucan (BDG) assay provides a reliable marker for the diagnosis of IFIs in adults with haematological malignancies. We assessed the diagnostic performance of this test in neonatal IYIs and its contribution to the monitoring of antifungal treatment. A retrospective study was performed in the NICU of the French University Hospital of Amiens from February 2012 to February 2014. Forty-seven neonates (33 males, 14 females) with a median gestational age of 30 weeks (IQR: 27-31) and median birth weight of 1200 g (IQR: 968-1700) were included and divided into three groups: 21 control neonates (CTRL), 20 neonates with probable IYI (PB), and six with proven IYI (PV). Median BDG levels were significantly higher in the global IYI group (PB + PV): 149 pg/ml (IQR: 85-364) vs. CTRL group: 39 pg/ml (IQR: 20-94) (P < .001). The optimal cut-off was 106 pg/ml (sensitivity 61.5%; specificity 81%). BDG levels decreased with antifungal treatment. BDG was detectable in cerebrospinal fluid, but the interest of this for diagnostic purposes remains unclear. Our results suggest that the BDG assay may be useful for the early identification of IYIs in neonates and for monitoring antifungal therapy efficacy.


Nicotinic acetylcholine receptor ion channel blockade by cocaine: the mechanism of synaptic action.
The action of cocaine on neuromuscular transmission of the frog has been studied to unveil the molecular site upon which this agent acts to block synaptic function. Indirectly elicited twitches of the sciatic nerve-sartorius muscle were reduced at greater than or equal to 50 microM cocaine. The electrically evoked action potentials of muscle membrane were inhibited at 200 microM cocaine, in a manner consistent with a local anesthetic effect. At the synaptic region, the quantal release of transmitter from the nerve terminal was not affected by cocaine at concentrations up to 100 microM. The most potent action of cocaine was the blockade of the ion channel of the nicotinic acetylcholine receptor. Transient membrane depolarization induced by microiontophoresis of acetylcholine was reduced to 7% of control at 50 microM cocaine in a dose-related manner; receptor desensitization was affected to a lesser extent by cocaine at all concentrations tested. The decay time constants of endplate currents were reduced at concentrations greater than or equal to 10 microM. The single channel conductances remained unchanged whereas the channel lifetimes were greatly reduced at 25 and 50 microM of the drug. Unblocking could not be discerned from the observed kinetics of either endplate current decay or single channel closed durations. However, with a combination of 10 microM cocaine and 0.4 M ethanol the endplate currents became biphasic. This suggested a blocking of open ion channels of the nicotinic receptor by cocaine followed by dissociation of acetylcholine from the blocked channel (in the absence of ethanol).


MR imaging of the lymphatic system: distribution and contrast enhancement of gadodiamide after intradermal injection.
We examined 24 lower extremities in 12 patients with lymphedema to evaluate the distribution and enhancement of gadodiamide after intradermal injection for MR imaging of lymphatic pathways in lymphedematous patients. The lymphedema was bilateral in 8, unilateral in 4, and 3 patients suffered from lymphedema in the genital region. 9 mL of gadodiamide and 1 mL of mepivacainhydrochloride 1% were divided into 5 portions and injected intradermally into the dorsal aspect of each foot. For MR imaging, a 3D spoiled gradient-echo sequence (Volumetric Interpolated Breathold Examination, VIBE) was performed. We detected the beaded appearance of lymphatic vessels extending from the injection site in 22 lower extremities (92%). In 13 lower extremities (54%), lymphatic vessels of the upper leg could be visualized. A contrast enhancement was observed in 16 out of 24 inguinal lymph node groups (67%). After 15 minutes of contrast material application, concomitant venous enhancement was detected in all lower extremities (100%). In 15 lower extremities (63%), collateral vessels with dermal backflow areas between lymphatic vessels were seen. Thus, intradermal injection of gadodiamide allows the visualization of lymphatic pathways in patients with lymphedema. In comparison to the venous system, lymphatic vessels show a tendency to have the highest contrast material uptake in the later acquisitions of 35, 45, and 55 minutes after intradermal injection of gadodiamide. Furthermore, 3D MIP reconstructions supported the identification of the lymphatic vessels and differentiation from veins due to the different angles of view.


Modes of De-identification.
De-identification of protected health information is an essential method for protecting patient privacy. Most institutes require de-identification of patient data prior to conducting scientific studies; therefore, it is important for clinical scientists to be cognizant of all modes of de-identification and all services provided by their de-identification tools. In this article, we discuss eight different modes of de-identification that yield de-identified data at different levels of quality. Most of these modes can be used in combination to achieve the best performance.


Oblique facial clefts: case report.
In a material of about 3,600 facial clefts eleven oblique clefts were observed (3.1%). The most frequent types were the medial oro-ocular and naso-ocular clefts and their combinations. There were two atypical cases and one lateral oro-ocular variety. The oblique clefts were frequently associated with other types of clefts and/or more distant anomalies. No genetic chromosomal or environmental factors were found to explain their etiology. Amniotic bands could possibly be thought to be responsible for the severe malformations in two instances. The lacrimal apparatus was always involved to some degree with the exception of a few incomplete forms. In the alveolar arch the cleft seemed to be situated between the medial incisor and the canine.


An Automated Immunoblot Method for Detection of IgG Antibodies to Hepatitis C Virus: a Potential Supplemental Antibody Confirmatory Assay.
An estimated 41,200 people were newly infected with hepatitis C virus (HCV) in 2016 in the United States. Screening tests for antibodies to HCV may generate up to 32% false positivity in low-risk populations. Current Centers for Disease Control and Prevention (CDC) screening recommendations do not require confirmatory testing of a screening anti-HCV-positive test; however, confirmation is valuable for surveillance in the absence of HCV RNA testing. A recombinant immunoblot assay (RIBA) was used as a confirmatory assay for anti-HCV-reactive samples but was discontinued in 2013. Another anti-HCV confirmatory assay, INNO-LIA, is commercially available in Europe but is not approved by the Food and Drug Administration (FDA) in the United States. We report the development of an anti-HCV assay that was performed on an automated immunoblot platform using a fourth-generation HCV recombinant fusion protein. Based on testing of 70 well-characterized samples, of which 40 were HCV RNA and anti-HCV positive, 15 were HCV RNA positive/anti-HCV negative, and 15 were HCV RNA and anti-HCV negative, the specificity and sensitivity of the HCV-WES assay were 100% and 95%, respectively. Concordance between INNO-LIA and HCV-WES was determined by testing 205 HCV RNA-negative/anti-HCV-positive samples, of which 149 (72.7%) were positive by HCV-WES, while 146 (71.2%) were positive by INNO-LIA. We have shown proof of concept for the use of this test for confirmation of screened anti-HCV results. The HCV-WES assay has advantages over manual Western blot assays and INNO-LIA, including ease of use, lower cost, and reduced hands-on time.


Frequency of family meals and childhood overweight: a systematic review.
Energy balance is influenced by understudied genetic, social and other environmental factors. The frequency of family meals (FFM) may be one of these factors since it is associated with a healthier dietary pattern in children and adolescents. The objective of this review is to evaluate the scientific evidence on the association between FFM and the risk of childhood and adolescent overweight. The electronic literature search identified 394 articles published during 2005-2012. Of these, 15 studies gave precise information of the studied association, of which four were longitudinal. We found great variability regarding the measurement of FFM. Six out of 11 cross-sectional studies and 1 out of 4 longitudinal studies found statistically significant inverse associations between FFM and being overweight, mainly in children, with odds ratios ranging from 0.11 to 0.93. Of those, only one adjusted for all the potential confounding factors considering socio-demographic, physical activity- and diet-related variables. Therefore, this review found inconsistent and weak evidence of an inverse association between FFM and risk of childhood overweight. In conclusion, further research is needed to establish whether family meals have an effect on childhood overweight. These studies ideally should have longitudinal or experimental designs, a clear and standardized definition of the exposure under study, a measure of the exposure based on direct observation or validated questionnaires and an adequate adjustment for potential confounders.


Synthesis and Characterization of Curcumin Incorporated Multi Component Nano-Scaffold with Enhanced Anti-bacterial and Wound Healing Properties.
Wound healing is one of the major challenges in chronic diseases; the current treatment options are less effective with undesirable side effects and are expensive. Extensive research is carried out to develop cost-effective, natural, biodegradable wound dressings that can reduce oxidative stress and inflammation and prevent bacterial infections. Curcumin has a plethora of therapeutic applications; however, its low solubility limits its clinical use. In this study, curcumin nanoparticles (Cur NP) and curcumin-chitosan nanoparticles (CCNP) were incorporated into the chitosan collagen vanillin scaffold, characterized, and investigated their potential wound healing properties. The nano-scaffolds were prepared by freeze-drying method and were characterized using Fourier transform infrared spectroscopy, X-ray diffraction, nanoparticle tracking analysis, and scanning electron microscopy. The drug release, antioxidant, antibacterial, and wound healing properties were assessed by in vitro assays. Cur nano-scaffolds showed particle sizes of 195.9 nm and 110.6 nm for Cur NP+VC and CCNP+VC, respectively. The curcumin encapsulated in the Cur NP+VC and CC+VC nano-scaffolds showed a release profile of > 60% and an improved antioxidant activity of greater than 80%. The nanoscaffolds were antagonistic against Escherichia coli and Staphylococcus aureus and enhanced wound healing capacity of 85.62 % and 77.05% in the murine cell line. The curcumin nano-scaffold is a biodegradable and effective drug delivery system for topical use that can act as an antioxidant, facilitate wound healing, as well as prevent bacterial infections.


Ultrasound-guided needle handling using a guidance positioning system in a phantom.
The SonixGPS™ needle guidance positioning system provides navigation assistance to facilitate needle handling during ultrasound-guided procedures. Each of 20 inexperienced nurse anaesthetists performed 12 different ultrasound-guided tasks in a porcine phantom. Using both in-plane and out-of-plane approaches, they inserted a needle and made contact with metal rods at depths of 2, 4 and 6 cm. We compared their performances without and with navigation as paired observations. Using the out-of-plane approach, navigation yielded shorter execution times (26 s vs 14 s, respectively; p = 0.01) and fewer needle repositionings (8 vs 3, respectively; p = 0.001). Using the in-plane approach, the needle was more visible with navigation assistance: 24% vs 52% of execution time, respectively (95% CI: 44%-12%; p = 0.0025). Better needle visibility was associated with shorter execution times and fewer needle repositionings. Combining ultrasound-guided techniques with the needle guidance positioning system may reduce tissue manipulation, thus improving patient comfort and safety.


Tapia's syndrome after arthroscopic shoulder stabilisation under general anaesthesia and LMA.
Anaesthetic complications, albeit rare, still occur and may be severe and unanticipated, with significant morbidity. Extracranial ipsilateral palsy of the recurrent laryngeal and the hypoglossal nerves is known as the Tapia's syndrome. Damage to these nerves may result from displacement of the head during mask ventilation, endotracheal intubation, bronchoscopy or the use of a laryngeal mask airway (LMA). We describe unilateral paralysis of the muscles of the tongue and ipsilateral vocal cord due to a lesion of cranial nerves X and XII that occurred following LMA anaesthesia combined with plexus block. A 57-year-old man with a rupture of the right shoulder underwent arthroscopic shoulder stabilisation and internal fixation. General anaesthesia with aLMA was combined with an interscalene plexus block. After induction with propofol and fentanyl, a LMA was inserted with some difficulty without muscle relaxation. The cuff was inflated with 30 mL of air and further volumes of air until a "just-seal" pressure was obtained. The anaesthesia was maintained with sevoflurane in oxygen/air. The procedure was carried out in a semi-supine position with the head inclined slightly forward, and the upper body slightly elevated. Surgery lasted 55 min and anaesthesia 70 min. After surgery, the patient quickly regained consciousness and the LMA was removed when he was responding to commands and was able to fully open his mouth. During the immediate postoperative period, the patient's voice was hoarse but he breathed without difficulty. The following day, he developed dysphagia and slurred speech; on examination, paralysis of the left side of the tongue was found. The diagnosis of an acute injury to the hypoglossal and laryngeal recurrent nerves was made and the patient was transferred to the neurology clinic for further treatment. This rare complication reminds us not only of the importance of positioning during anaesthesia and surgery, but also of the need for careful and correct airway management. It could be probably prevented by careful insertion of an appropriate size LMA, and the use of low intracuff pressures and/or volumes.


Enhanced amplitude modulations contribute to the Lombard intelligibility benefit: Evidence from the Nijmegen Corpus of Lombard Speech.
Speakers adjust their voice when talking in noise, which is known as Lombard speech. These acoustic adjustments facilitate speech comprehension in noise relative to plain speech (i.e., speech produced in quiet). However, exactly which characteristics of Lombard speech drive this intelligibility benefit in noise remains unclear. This study assessed the contribution of enhanced amplitude modulations to the Lombard speech intelligibility benefit by demonstrating that (1) native speakers of Dutch in the Nijmegen Corpus of Lombard Speech produce more pronounced amplitude modulations in noise vs in quiet; (2) more enhanced amplitude modulations correlate positively with intelligibility in a speech-in-noise perception experiment; (3) transplanting the amplitude modulations from Lombard speech onto plain speech leads to an intelligibility improvement, suggesting that enhanced amplitude modulations in Lombard speech contribute towards intelligibility in noise. Results are discussed in light of recent neurobiological models of speech perception with reference to neural oscillators phase-locking to the amplitude modulations in speech, guiding the processing of speech.


[The architectonics of fetal placental veins in second-trimester cytomegalovirus infection].
To study a change in the architectonics of fetal surface veins of the placenta and the structure of the latter in cytomegalovirus infection in pregnant women. Placentas were studied and divided in 3 groups: 1) 35 placentas from women with physiological pregnancy; 2) 37 placentas from women with an exacerbation of latent cytomegalovirus infection and with chronic compensated placental insufficiency; 3) 30 placentas from those with an exacerbation of latent cytomegalovirus infection and with chronic subcompensated placental insufficiency. After X-ray contrasting the fetal surface veins of the placenta and collecting its sections for morphological analysis, the changes in the blood vessels were compared with the structure of the villous chorion. As compared with Group 2, Group 3 showed reductions in placental weights and placental areas, as well as cotyledon asymmetry and a preponderance of anatomical forms with weak vascular contrasting along with the more frequent emergence of areas of varicosity, contraction, and inflammation of the umbilical vein, as well as the placental fetal surface veins. The investigators more often detected collagenization, fibrinoid degradation, and the increased stromal volume of the stem villi, fibrinoid deposition around the latter, and decreases in the proportion of the intervillous space and syncytiotrophoblast; dilatation of the veins and narrowing of the arteries of the stem villi; pronounced plethora of terminal villi and their avascular forms, intermediate immature villi, chorioamnionitis, deciduitis, pseudonecrosis, calcifications, as well as hemorrhages in the intervillous space. In the development of subcompensated placental insufficiency in women who have experienced an exacerbation of latent cytomegalovirus infection in the second trimester of pregnancy, an important role is assigned to anatomical and pathological changes in the umbilical, fetal surface, and villous chorionic veins, which indicate the development of fetoplacental hypertension, the deceleration of blood flow, and the longer contact of the pathogen with the endothelium and syncytiotrophoblast.


Synthesis of the scFv fragment of anti-Frizzled-7 antibody and evaluation of its effects on triple-negative breast cancer in vitro study.
Among the most promising antibody formats in terms of inhibiting carcinogenesis are single-stranded variable fragments, whose targeted binding to the Fzd7 receptor has been proven effective at suppressing tumorigenesis. In this study, we investigated the effectiveness of an anti-Fzd7 antibody fragment against both tumor growth and metastasis of breast cancer cells. To develop anti-Fzd7 antibodies, bioinformatics approaches were used and the antibodies were expressed recombinantly in E. coli BL21 (DE3). The expression of anti-Fzd7 fragments was verified by Western blotting. Analysis of the antibody's binding capacity to Fzd7 was conducted by flow cytometry. Cell death and apoptosis were assessed by MTT and Annexin V/PI assays. The transwell migration and invasion assays, as well as the scratch method, were used to evaluate cell motility and invasiveness. The anti-Fzd7 antibody was expressed successfully as a single band of 31 kDa. It could bind to 21.5% of MDA-MB-231 cells, as opposed to only 0.54% of SKBR-3 cells as negative control. According to MTT assay, induced apoptosis was 73.7% in MDA-MB-231 cells, compared with 29.5% in SKBR-3 cells. Also, the antibody exerted a significant inhibitory effect of 76% and 58% on migration and invasion of MDA-MB-231 cells, respectively. The recombinantly developed anti-Fzd7 scFv of this study could exhibit significant antiproliferative and antimigratory properties, along with a high apoptosis-inducing potential, making it suitable for the immunotherapy of triple negative breast cancer.


Saving for hospital care costs in Singapore: are there lessons for Australia? [corrected].
Australian political leaders and policy analysts have increasingly sought to learn from Singapore, a nation which, despite its small size and lack of natural resources, has enjoyed rapid economic growth. Of particular interest to Australian policymakers has been Singapore's compulsory superannuation system, which has provided high levels of domestic savings and high levels of home ownership; it has also incorporated a scheme to enable Singaporeans to save for the costs of health care. In this article, Singapore's Medisave policy is described within its broader socioeconomic context. Saving for health care costs has been a largely neglected option in recent policy debate about reforming funding arrangements for hospital care in Australia. The potential for a formalised scheme for medical savings in Australia, subsidised by taxation concessions, is explored in terms of its socioeconomic policy implications, its congruence with Australian values, and the logistical ramifications of such a scheme.


Neurological and psychiatric presentations associated with COVID-19.
The objective is to investigate coronavirus disease 2019 (COVID-19)-associated neurological and psychiatric effects and explore possible pathogenic mechanisms. This study included 77 patients with laboratory-confirmed COVID-19 in Wuhan, China. Neurological manifestations were evaluated by well-trained neurologists, psychologists, psychiatric presentations and biochemical changes were evaluated using the Generalized Anxiety Disorder 7-item scale, Patient Health Questionnaire-9, Brief Psychiatric Rating Scale, and electronic medical records. Eighteen (23.4%) patients presented with neurological symptoms. Patients with neurological presentations had higher urea nitrogen, cystatin C, and high-sensitivity C-reactive protein levels and lower basophil counts. Among them, patients with muscle involvement had higher urea nitrogen and cystatin C levels but lower basophil counts. In addition, patients with psychiatric presentations were older and had higher interleukin (IL)-6 and IL-10 levels and higher alkaline phosphatase, R-glutamate transferase, and urea nitrogen levels. Moreover, patients with anxiety had higher IL-6 and IL-10 levels than those without, and patients with moderate depression had higher CD8 + T cell counts and lower CD4 + /CD8 + ratios than other patients. This study indicates that the central nervous system may be influenced in patients with COVID-19, and the pathological mechanisms may be related to direct virus invasion of the central nervous system, infection-mediated overreaction of the immune system, and aberrant serum pro-inflammatory factors. In addition, basophils and cystatin C may also play important roles during these pathological processes. Our findings suggest that neurological and psychiatric presentations should be evaluated and managed in patients with COVID-19. Further studies are needed to investigate the underlying mechanisms.