Protein aggregates as depots for the release of biologically active compounds.
Protein misfolding and aggregation is one of the most serious problems in cell biology, molecular medicine, and biotechnology. Misfolded proteins interact with each other or with other proteins in non-productive or damaging ways. However, a new paradigm arises that protein aggregation may be exploited by nature to perform specific functions in different biological contexts. From this consideration, acceleration of stress-induced protein aggregation triggered by any factor resulting in the formation of soluble aggregates may have paradoxical positive consequences. Here, we suggest that amorphous aggregates can act as a source for the release of biologically active proteins after removal of stress conditions. To address this concept, we investigated the kinetics of thermal aggregation in vitro of yeast alcohol dehydrogenase (ADH) as a model substrate in the presence of two amphiphilic peptides: Arg-Phe or Ala-Phe-Lys. Using dynamic light scattering (DLS) and turbidimetry, we have demonstrated that under mild stress conditions the concentration-dependent acceleration of ADH aggregation by these peptides results in formation of large but soluble complexes of proteins prone to refolding.


The role of NHE-1 in myocardial hypertrophy and remodelling.
Na-H exchange (NHE) is the primary process by which the cardiac cell extrudes protons particularly under conditions of intracellular acidosis. Nine isoforms of NHE have now been identified. Although these antiporters are expressed in virtually all tissues, cardiac cells posses primarily the ubiquitous NHE-1 subtype. It has been well established that NHE-1 is a major contributor to acute ischemic and reperfusion injury although it is now emerging that NHE-1 contributes to chronic maladaptive myocardial responses to injury such as post-infarction myocardial remodelling and likely contributes to the development of heart failure. Experimental studies using both in vitro approaches as well as animal models of heart failure have consistently demonstrated a beneficial effect of NHE-1 inhibitors in attenuating hypertrophy in response to various stimuli as well as inhibiting heart failure in a variety of animal models representing experimentally-induced or genetic models of heart failure. The beneficial effects of NHE-1 inhibitors occur independently of infarct size reduction or on any direct effects on afterload thus implicating a direct antiremodelling influence of these agents. It is proposed that NHE-1 inhibition represents a potentially effective new therapeutic approach for the treatment of heart failure.


Growth hormone and cardiovascular risk markers.
The association of hypopituitarism and growth hormone (GH) deficiency with increased cardiovascular mortality has become increasingly well established. In GH-deficient adults, whether the onset of disease is in childhood or in adult life, a higher prevalence of atherosclerotic plaques and endothelial dysfunction has been described. Of note, high-resolution ultrasonography has shown that patients with hypopituitarism have a higher prevalence of premature atherosclerosis even in the absence of clinical symptoms. Specific anatomic abnormalities also have been described in adults with GH deficiency, including an increase in intimal-medial wall thickness. The contribution of specific hormonal factors and prior irradiation to the development of vascular disease remains controversial.


STRA6 exerts oncogenic role in gastric tumorigenesis by acting as a crucial target of miR-873.
Increasing evidence shows that stimulated by retinoic acid 6 (STRA6) participates in regulating multiple cancers. However, the biological roles of STRA6 in gastric cancer (GC) remain unknown. This study aimed to investigate the biological function of STRA6 and reveal the underlying mechanism of its dysregulation in GC. The expression level of STRA6 was detected through quantitative real-time PCR and Western blot analysis. The effects of STRA6 on the proliferation of GC cells were studied through CCK-8 proliferation, colony formation and 5-ethynyl-2'-deoxyuridine (EdU) assays. The effects of STRA6 on migration and invasion were detected via wound healing and Transwell assays. Upstream miRNAs, which might regulate STRA6 expression, was predicted through bioinformatics analysis. Their interaction was further confirmed through dual-luciferase reporter assays and rescue experiments. STRA6 was up-regulated in GC and enhanced the proliferation and metastasis of GC cells in vitro and in vivo. STRA6 knockdown could inhibit the Wnt/β-catenin signalling pathway. STRA6 was confirmed as an miR-873 target, which acted as a tumour suppressor in GC. Rescue assays showed that the repressing effect of miR-873 could be partially reversed by overexpressing STRA6. STRA6 is down-regulated by miR-873 and plays an oncogenic role by activating Wnt/β-catenin signalling in GC.


Primary and secondary control over eating behaviors.
The relationships between subjective control, body image, and eating behaviors were examined within the framework of the Optimization in Primary and Secondary Control model (OPS model; Heckhausen, 1999). This model characterizes control as an adaptive and strategic process in which the target of control includes internal as well as external states, and in which the purpose is either to facilitate goal pursuit by engaging with the goal or managing the consequences of goal failure by disengaging from the goal. A convenience sample of 180 Australian women (age: M=26.49, SD=5.03) completed the Optimization of Primary and Secondary Control Scale (OPS scale Heckhausen, 1999), as well as measures of attitudinal and behavioral factors comprising Stice's (1994) dual-pathway model of bulimia. A series of path analyses revealed that the control strategies involved in goal engagement are directly associated with increased dietary restraint and purging, whereas the control strategies involved in goal disengagement are indirectly associated with these factors, and with negative affect, by way of reduced body dissatisfaction and reduced frequency of appearance comparisons. These results suggest that goal engagement strategies, which are typically adaptive in other contexts, are associated with potentially hazardous attitudes and behaviors in the context of the body. The results also suggest that an ability to disengage from body weight goals is associated with a reduced likelihood of developing disordered eating.


Guiding the design space for nanotechnology to advance sustainable crop production.
The globally recognized need to advance more sustainable agriculture and food systems has motivated the emergence of transdisciplinary solutions, which include methodologies that utilize the properties of materials at the nanoscale to address extensive and inefficient resource use. Despite the promising prospects of these nanoscale materials, the potential for large-scale applications directly to the environment and to crops necessitates precautionary measures to avoid unintended consequences. Further, the effects of using engineered nanomaterials (ENMs) in agricultural practices cascade throughout their life cycle and include effects from upstream-embodied resources and emissions from ENM production as well as their potential downstream environmental implications. Building on decades-long research in ENM synthesis, biological and environmental interactions, fate, transport and transformation, there is the opportunity to inform the sustainable design of nano-enabled agrochemicals. Here we perform a screening-level analysis that considers the system-wide benefits and costs for opportunities in which ENMs can advance the sustainability of crop-based agriculture. These include their on-farm use as (1) soil amendments to offset nitrogen fertilizer inputs, (2) seed coatings to increase germination rates and (3) foliar sprays to enhance yields. In each analysis, the nano-enabled alternatives are compared against the current practice on the basis of performance and embodied energy. In addition to identifying the ENM compositions and application approaches with the greatest potential to sustainably advance crop production, we present a holistic, prospective, systems-based approach that promotes emerging alternatives that have net performance and environmental benefits.


Monoacylglycerols activate capsaicin receptor, TRPV1.
Transient receptor potential vanilloid subtype 1 (TRPV1) is known as capsaicin (CAP) receptor and activated by CAP. Activation of TRPV1 by CAP increases energy expenditure and thermogenesis in rodents or human. Therefore, TRPV1 may be target for energy expenditure enhancement and thermogenesis. To search for novel TRPV1 agonist, we screened 19 types of foods by using TRPV1-expressing HEK293 cells. TRPV1 was activated by hexane extract of wheat flour, and its functional compounds were 1-monoacylglycerols containing oleic, linoleic, and alpha-linolenic acids. Their potencies (EC50) were about 50 times larger than that of CAP and their efficacies (maximal response) were about half of that of CAP. TRPV1 was activated by 1-monoacylglycerols (MGs) having C18 and C20 unsaturated and C8-C12 saturated fatty acid (FA). Moreover, 2-MGs having C18 and C20 unsaturated FA acted on TRPV1 with the same potency. On the other hand, no activation of TRPV1 was induced by MGs having C16 and C18 saturated FA, di- or triacylglycerols of C18:1 FA. Pain-relating aversive responses were induced when TRPV1-activating 1-monoacylglycerols (50 mM) was administered subcutaneously into rat hind paw. These effects were inhibited by the co-injection of capsazepine (10 mM) which is a TRPV1 competitive antagonist. These results suggested that these 1-monoacylglycerols activate TRPV1 in vitro and in vivo.


Standardization of spinal manipulation therapy in humans: development of a novel device designed to measure dose-response.
The main objective of this report is to present an innovative research tool that will provide the opportunity to study fundamental aspects of the spinal manipulation dose-physiological response relation in humans. A servo-controlled linear actuator motor was developed to simulate spinal manipulative therapy. Coefficient of multiple correlations was calculated to assess the degree of similarity between each measured force curves, whereas precision was obtained by comparing resulting peak force and time-to-peak force to the target curves. The coefficient of multiple correlations calculations showed that repeatability was very high with all correlation values over 0.98. Precision was also very high with average differences in peak force and time-to-peak force of less than 3 N and less than 5 milliseconds. The tool was designed to optimize precision, repeatability, and safety in the delivery of force to the spine in humans. It offers a unique opportunity to study dose-response relationship for several spinal manipulation parameters such as peak force, time-to-peak force, and preload.


Prostaglandins in fetal tracheal and amniotic fluid from late pregnant sheep.
Concentrations of prostaglandin E (PGE), prostaglandin F (PGF) and 13,14-dihydro-15-keto-prostaglandin F (PGFM) have been measured in fetal tracheal and amniotic fluid from chronically catheterized sheep during late pregnancy. Amniotic fluid contained significantly greater concentrations of these prostaglandins than tracheal fluid (p less than 0.01); there was no correlation between the level of prostaglandins found in each fluid. In tracheal fluid concentrations of PGE and PGFM exceeded those of PGF (P less than 0.01) whereas no significant differences were found in amniotic fluid. The levels of prostaglandins in these fluids were similar in ewes bearing hypophysectomized fetuses.


Characterization of a CTX-M-15 Producing Klebsiella Pneumoniae Outbreak Strain Assigned to a Novel Sequence Type (1427).
Extended-spectrum -lactamase producing Klebsiella pneumoniae have emerged as one of the major nosocomial pathogens. Between July and September 2012, a CTX-M-15 producing K. pneumoniae caused an outbreak in a university hospital in the Netherlands. The outbreak isolates were characterized and assigned to a novel sequence type (ST1427). An epidemiological link between affected patients was supported by patient contact tracing and whole-genome phylogenetic analysis. Intra-strain polymorphism was detected among multiple isolates obtained from different body sites of the index patient, which may relate to antibiotic treatment and/or host adaptation. Environmental contamination caused by the outbreak clone was found in the patient rooms even on medical equipment. The novel clone was not closely related to any known endemic/epidemic clone, but carried a set of a plasmid-borne resistance genes [bla CTX-M-15, bla TEM-1, bla OXA-1, aac(6')-Ib-cr, qnrB1, tetA(A), aac(3)-II]. Analysis of its virulence factors revealed a previously uncharacterized capsular biosynthesis region and two uncharacterized fimbriae gene clusters, and suggested that the new clone was not hypervirulent. To our knowledge, this is the first outbreak report of K. pneumoniae ST1427, and our study could be of help to understand the features of this newly emerging clone.


RanGAP mediates GTP hydrolysis without an arginine finger.
GTPase-activating proteins (GAPs) increase the rate of GTP hydrolysis on guanine nucleotide-binding proteins by many orders of magnitude. Studies with Ras and Rho have elucidated the mechanism of GAP action by showing that their catalytic machinery is both stabilized by GAP binding and complemented by the insertion of a so-called 'arginine finger' into the phosphate-binding pocket. This has been proposed as a universal mechanism for GAP-mediated GTP hydrolysis. Ran is a nuclear Ras-related protein that regulates both transport between the nucleus and cytoplasm during interphase, and formation of the mitotic spindle and/or nuclear envelope in dividing cells. Ran-GTP is hydrolysed by the combined action of Ran-binding proteins (RanBPs) and RanGAP. Here we present the three-dimensional structure of a Ran-RanBP1-RanGAP ternary complex in the ground state and in a transition-state mimic. The structure and biochemical experiments show that RanGAP does not act through an arginine finger, that the basic machinery for fast GTP hydrolysis is provided exclusively by Ran and that correct positioning of the catalytic glutamine is essential for catalysis.


Real-time 3-dimensional contrast-enhanced ultrasound in detecting hemorrhage of blunt renal trauma.
The objective of this study is to evaluate the diagnostic value of real-time 3-dimensional contrast-enhanced ultrasound in the hemorrhage of blunt renal trauma. Eighteen healthy New Zealand white rabbits were randomly divided into 3 groups. Blunt renal trauma was performed on each group by using minitype striker. Ultrasonography, color Doppler flow imaging, and contrast-enhanced 2-dimensional and real-time 3-dimensional ultrasound were applied before and after the strike. The time to shock and blood pressure were subjected to statistical analysis. Then, a comparative study of ultrasound and pathology was carried out. All the struck kidneys were traumatic. In the ultrasonography, free fluid was found under the renal capsule. In the color Doppler flow imaging, active hemorrhage was not identified. In 2-dimensional contrast-enhanced ultrasound, active hemorrhage of the damaged kidney was characterized. Real-time 3-dimensional contrast-enhanced ultrasound showed a real-time and stereoscopic ongoing bleeding of the injured kidney. The wider the hemorrhage area in 4-dimensional contrast-enhanced ultrasound was, the faster the blood pressure decreased. Real-time 3-dimensional contrast-enhanced ultrasound is a promising noninvasive tool for stereoscopically and vividly detecting ongoing hemorrhage of blunt renal trauma in real time.


Mild Synthesis of Pt/SnO2 /Graphene Nanocomposites with Remarkably Enhanced Ethanol Electro-oxidation Activity and Durability.
We have designed a new Pt/SnO2 /graphene nanomaterial by using L-arginine as a linker; this material shows the unique Pt-around-SnO2 structure. The Sn(2+) cations reduce graphene oxide (GO), leading to the in situ formation of SnO2 /graphene hybrids. L-Arginine is used as a linker and protector to induce the in situ growth of Pt nanoparticles (NPs) connected with SnO2 NPs and impede the agglomeration of Pt NPs. The obtained Pt/SnO2 /graphene composites exhibit superior electrocatalytic activity and stability for the ethanol oxidation reaction as compared with the commercial Pt/C catalyst owing to the close-connected structure between the Pt NPs and SnO2 NPs. This work should have a great impact on the rational design of future metal-metal oxide nanostructures with high catalytic activity and stability for fuel cell systems.


What Counts as Disease?. Rationales and Rationalizations for Treatment.
Disease is a fact of nature. Diagnosis is an artefact constructed by human beings. The core concept of disease is a bodily abnormality. Literally, the term 'disease' denotes a demonstrable lesion of cells, tissues, or organs; metaphorically, it may be used to denote any kind of malfunctioning, of individuals, groups, economies. Classic nosology was descriptive, based on somatic pathology. The diagnostician sought to anticipate and approximate the pathologist's findings at autopsy, that is, identify the patients's bodily lesion/disease and its material cause (etiology). For example, the term 'pneumococcal pneumonia' identifies the organ affected, the lungs, and the cause of the illness, infection with the pneumococcus. Contemporary nosology is strategic, based on economic, legal, social, and other interests (unrelated to disease as somatic pathology). The diagnostician seeks to secure reimbursement for medical services, legitimize treatment, justify defining undesirable behavior as disease, and so forth. For example, diagnosis-related groups provide bureaucratic rationale for reimbursing medical services by third-party payers; psychiatric diagnoses provide legal-scientific rationale for treating mental diseases as if they were brain diseases; and so forth. Formerly, diagnoses encoded the objectively verifiable condition of the patient's body (diseases). Today, diagnoses rationalize the health-care policy of the body politic (methods of controlling costs and compensating physicians). We are witnessing the transformation of nosology from the medical-scientific classification of disease as somatic pathology, into the medicalized justification of social policy as 'health care' or 'treatment'.


Spectral analysis of the electroencephalogram in the developing rat.
Power spectral measures of the EEG obtained from the frontal cortex and hippocampal formation during different vigilance states in the developing rat have been computed and compared. The most significant ontogenetic changes were observed in the hippocampal power spectra obtained during the vigilance state of REM sleep. These spectral analyses have revealed in the hippocampus: (1) a significant increase in the frequency at which the peak power occurs in the theta-frequency (4-11 Hz) band from 14 to 45 days of age; (2) a decrease in the quality factor of the peak from 14 to 45 days of age; (3) a decrease in the relative power co-ordinate for the center of spectral mass associated with the 0-4-Hz frequency band coupled with an increase in the frequency coordinate of the 4-11-Hz frequency band from 14 to 45 days of age, and; (4) a significant decrease in the average percent relative power associated with the 0-4-Hz frequency band from 14 to 22 days of age. For the EEG obtained from the frontal cortex, the major findings of note were: (1) a dominant contribution of relative power in the 0-4-Hz frequency band which was observed at every age and during every vigilance state tested, and; (2) a significant increase in the average percent relative power associated with this band at 18, 22, and 45 days of age. The results of this study provide a quantitative description of the electroencephalographic (EEG) ontogeny of the hippocampal formation and the frontal cortex in the rat. These ontogenetic changes in EEG activity relate closely to development of the internal circuitry and synaptic maturation in the hippocampal formation and frontal cortex.


Mathematical Modelling of Residual-Stress Based Volumetric Growth in Soft Matter.
Growth in nature is associated with the development of residual stresses and is in general heterogeneous and anisotropic at all scales. Residual stress in an unloaded configuration of a growing material provides direct evidence of the mechanical regulation of heterogeneity and anisotropy of growth. The present study explores a model of stress-mediated growth based on the unloaded configuration that considers either the residual stress or the deformation gradient relative to the unloaded configuration as a growth variable. This makes it possible to analyze stress-mediated growth without the need to invoke the existence of a fictitious stress-free grown configuration. Furthermore, applications based on the proposed theoretical framework relate directly to practical experimental scenarios involving the "opening-angle" in arteries as a measure of residual stress. An initial illustration of the theory is then provided by considering the growth of a spherically symmetric thick-walled shell subjected to the incompressibility constraint.


Effects of electrical stimulation and reversible lesions of the olivocerebellar pathway on Purkinje cell activity in the flocculus of the cat.
The activity of Purkinje cells (P-cells) in the flocculus of 8 lightly anesthetized cats and one alert cat was recorded for periods of up to several hours each. The resting simple spike (SS) rate in the anesthetized cats was 37 +/- 21 Hz (mean +/- standard deviation), similar to that observed in the alert cat. Complex spikes (CSs) were evoked by an electrode placed in the climbing fiber (CF) decussation or inferior olive (IO). For each P-cell, SS activity was suppressed completely at or above a cut-off frequency of evoked CSs; median cut-off rate was 5 Hz (range 1-10 Hz, 15 cells). Reversible lesions of the CF pathway were made by microinjection of 1-10 microliter of saturated lidocaine into the IO or CF decussation. This abolished spontaneous CS activity and produced two reversible effects on SS discharge: (1) an increase in mean SS rate of 98%, from 23 +/- 13 to 40 +/- 18 Hz (11 cells); and (2) a decrease of 50% in the variability of SS firing rate. Similar effects were observed in two P-cells whose CF axons were lesioned mechanically. These results show that electrical stimulation and reversible lesions in areas previously shown to alter the gain of the vestibulo-ocular reflex (VOR) also alter CF input to the flocculus, suggesting that the gain changes were caused by changes in CS rate. This study confirms and extends the observation that a reciprocal relationship exists between CS rate and SS background rate, and therefore further suggests that the changes in the gain of the VOR might be due to changes in SS background rate.


Distinguishing dengue fever from other infections on the basis of simple clinical and laboratory features: application of logistic regression analysis.
Dengue fever is a frequent cause of admission to hospital in South East Asia, however many of the clinical characteristics and abnormalities on laboratory investigations at presentation are found in other common infections. To describe the clinical and laboratory features of dengue fever and other common febrile illnesses in Singapore. We performed a prospective study of consecutive adult admissions to an infectious diseases hospital. Logistic regression analysis was used to identify symptoms, physical signs and laboratory features that differentiated dengue fever from other febrile illnesses within the first 2 days of admission. Of the 381 patients studied, 148 had serologically confirmed dengue fever. Most of these had uncomplicated dengue fever, and only 9% had dengue haemorrhagic fever. A model based on clinical features alone (including a variety of cutaneous signs, pulse rate and the presence of pharyngeal injection) was able to differentiate dengue fever from other infections with a sensitivity of 74% and specificity of 79%. A model based on clinical features (rash) and laboratory parameters (white cell count, haemoglobin, prothrombin time, creatinine and bilirubin levels), achieved a sensitivity of 84% and specificity of 85%. A combination of simple clinical and laboratory parameters are potentially able to predict dengue fever with a high level of accuracy in adults presenting to hospital with febrile illnesses in Singapore.


The lymphoproliferative disease of granular lymphocytes. A heterogeneous disorder ranging from indolent to aggressive conditions.
A multiparameter analysis, which included the evaluation of clinical features, cell morphology, karyotype, phenotypic and functional immunologic findings, and T-cell receptor beta-chain configuration was performed on 34 patients with lymphoproliferative disease of granular lymphocytes (LDGL). The two-fold aim of the study was to identify the most useful tools that would more accurately characterize these patients and to deal with the problem of classifying these lymphoproliferative disorders. The data presented in this article suggest that a single parameter may not be sufficient to define the nature of the proliferating cells or to predict the clinical course of the disease and prognosis for the patient. The use of a multiparameter approach, however, may reach this goal, thus providing important prognostic and therapeutic information. Our study supports the concept that lymphoproliferative disease of granular lymphocytes is a heterogeneous disorder that ranges from indolent and possibly reactive conditions to the manifestation of aggressive malignancies.


Botulinum toxin A for refractory OAB and idiopathic urinary retention: Can phenotyping improve outcome for patients: ICI-RS 2019?
Botulinum toxin A (BTX-A) is a well-established treatment for refractory idiopathic overactive bladder (OAB). It has also been used with short-term success in treating idiopathic urinary retention. However, efficacy and complication rates are variable and predicting those likely to benefit most from treatment would enable personalization of therapy and optimization of outcomes. At the International Consultation on Incontinence-Research Society (ICI-RS) meeting in 2019 a Think Tank addressed the question of how we can improve the way we phenotype patients undergoing BTX-A treatment. The Think Tank conducted a literature review and expert consensus meeting focussing on how advances in basic science research of the mechanism of action of BTX-A, as well as assessment of psychological comorbidity, can be translated into clinical practice to improve patient selection for therapy. Idiopathic OAB and idiopathic urinary retention are heterogenous conditions encompassing several phenotypes with multiple potential pathophysiological mechanisms. Animal models have demonstrated a central nervous system mechanism of action of intravesically injected BTX-A and this has been confirmed in human functional MRI studies, but whether this tool can be used to predict outcome from treatment remains to be determined. Phenotyping based on psychological comorbidity using validated screening tools should be studied as a way to potentially optimize patient selection for therapy. Advances in basic science research into the mechanism of action of BTX-A have improved our understanding of the pathophysiology of OAB and may lead to novel ways to phenotype patients. Psychological assessment is another way in which phenotyping may be improved. Areas for further research are proposed.


Oral intake of ZrO2 nanoparticles by pregnant mice results in nanoparticles' deposition in fetal brains.
Nanotoxicity to fetal brains after maternal oral exposures during pregnancy is often in question because nanoparticles have to cross multiple biological barriers such as intestinal barrier, maternal blood placental barrier (BPB) and fetal blood brain barrier (BBB). Here, we investigated this seemingly impossible passage for ZrO2 nanoparticles (ZrO2 NPs) from maternal body to fetal brains using a pregnant mouse model. After three oral exposures to pregnant mice at late pregnancy (GD16, 17, 18), ZrO2 NPs were able to accumulate in fetal brains at GD19 via crossing the well-developed maternal BPB and fetal BBB. Moreover, ZrO2 NPs crossed the mature biological barriers with increasing the expression levels of caveolae, clathrin and arf6 proteins as well as decreasing the expression levels of the tight junction proteins claudin-5, occludin and ZO-1 in placenta and fetal brain. From this investigation, we speculated that the main mechanisms for such translocation were receptor-mediated endocytosis transcellular pathway and breakthrough of tight junctions paracellular pathway in mature maternal BPB and fetal BBB. These findings have important implications for other nanoparticles exposures during pregnancy and provide crucial information to safeguard fetal development from contamination of widely used nanoproducts.


Development of a sensitive and quantitative diagnostic assay for fish nervous necrosis virus based on two-target real-time PCR.
The aim of the present work was to develop two new independent SYBR Green I-based real-time PCR assays for both detection and quantification of betanodavirus, an RNA virus that infects several species of marine teleost fish causing massive mortalities in larvae and juveniles. The assays utilized two pairs of primers targeting highly conserved regions of both the RNA molecules forming the betanodavirus genome: RNA1 encoding the RNA-dependent RNA polymerase (RdRP) and RNA2 encoding the coat protein (CP). The specificity of amplifications was monitored by the melting analysis and agarose gel electrophoresis of the amplified products. The applicability of these assays was confirmed with 21 betanodavirus strains, covering all the four main clades. In addition, a BLAST (NCBI) search with the primer sequences showed no genomic cross-reactivity with other viruses. The new assays were able to quantify concentrations of betanodavirus genes ranging from 10(1) to 10(8) copies per reaction. The intra-assay coefficients of variation (CV) of threshold cycle (Ct) values of the assays were 1.5% and 1.4% for CP and RdRP RNAs, respectively. The inter-assay CVs of Ct values were 2.3% and 2.4% for CP and RdRP RNAs, respectively. Moreover, regression analysis showed a significant correlation (R2>0.97) between genome number, as determined by real-time PCR assays and the corresponding virus titer expressed as TCID50/ml of two different betanodavirus strains propagated in cell culture. The two assays were compared with a previously established one-step RT-PCR assay and with the classical virus isolation test and found to be more sensitive. In conclusion, the developed real-time RT-PCR assays are a reliable, specific and sensitive tool for the quantitative diagnosis of betanodavirus.


Spatial and temporal influence of onsite wastewater treatment systems, centralized effluent discharge, and tides on aquatic hazards of nutrients, indicator bacteria, and pharmaceuticals in a coastal bayou.
In the rapidly urbanizing watersheds and estuaries flowing to the Gulf of Mexico in Texas, USA, instream flows are increasingly influenced by point source and nonpoint source discharges. Spatial and temporal tidal influences on water quality, especially for contaminants of emerging concern (CECs), is poorly understood in estuaries and coastal systems. We selected Dickinson Bayou, an urban estuary in Galveston County, Texas, for study because it has historically impaired water quality, receives point source discharge from one major wastewater treatment plant (WWTP), while also being influenced by high densities of onsite sewage facilities upstream in the watershed. We explored the occurrence and potential hazards of aquatic contaminants, including nutrients, indicator bacteria for pathogens, and CECs, in relation to this point source discharge, across seasons and at high and low tides. Aquatic contaminants and associated hazards varied significantly in relation to the WWTP discharge, and were influenced by onsite systems. In fact, spatiotemporal water quality varied by class of contaminants (e.g., nutrients, indicator bacteria, CECs), which indicates that traditional surface water monitoring activities should account for such environmental complexity. This study provides a diagnostic approach for future studies of emerging water quality challenges across gradients of rapidly urbanizing coastal bays and estuaries.


[Evaluation of the pseudocholinesterase activity in hyperlipoproteinemia type II and type IV].
Authors have evaluated pseudocholinesterase activity in patients with type IIa and type IV hyperlipoproteinemia. Significative correlation has been found between PCE and C in type IV hyperlipoproteinemia. Authors suggest PCE activity can be proposed as useful biochemical marker of hyperlipoproteinemia.


Derivation and validation of a new visceral adiposity index for predicting visceral obesity and cardiometabolic risk in a Korean population.
The visceral adiposity index (VAI), an indirect marker of visceral adipose tissue, serves as a model associated with cardiometabolic risk, but has limitations regarding the Asian population. We sought to develop a new VAI (NVAI) for the Korean population and compare it to VAI for prediction of atherosclerotic cardiovascular disease (ASCVD) risk and development of major cardiovascular diseases (CVD) and stroke. Patients (969) who underwent visceral fat area measurement were analyzed. After exclusion, 539 patients (142 men, 397 women) were randomly divided into internal (n = 374) and external validation (n = 165) data set. The NVAI was developed using univariate and multivariate logistic regression with backward selection of predictors. Receiver operating characteristic (ROC) curve analysis and comparison of the area under the curve (AUC) verified the better predictor of ASCVD risk score. Additionally, nationwide population-based cross-sectional survey data (Korean National Health and Nutrition Examination Survey [KNHANES] 2008-2010, n = 29,235) was used to validate the NVAI's ability to predict ASCVD risk and major CVD and stroke. The NVAI better reflected visceral fat area in internal and external data sets, with AUCs of 0.911 (95% confidence interval [CI]: 0.882-0.940) and 0.879 (95% CI: 0.828-0.931), respectively. NVAI better discriminated for ASCVD risk (AUC = 0.892, 95% CI: 0.846-0.938) compared to VAI (0.559, 95% CI: 0.439-0.679). The NVAI also better predicted MI or angina, and stroke with AUCs of 0.771 (95% CI: 0.752-0.789), and 0.812 (95% CI: 0.794-0.830), respectively, compared with waist circumference (WC), body mass index (BMI), TG to HDL ratio, and VAI via KNHANES, in a statistically significant manner. The NVAI has advantages as a predictor of visceral obesity and is significantly associated with ASCVD risks and development of major CVD and stroke in the Korean population. The NVAI could be a screening tool for improved risk estimation related to visceral obesity.


Efficacy and Safety of Switching to Dolutegravir/Lamivudine in Virologically Suppressed People Living with HIV-1 Aged Over 65 Years.
Clinical trials of dual regimen dolutegravir/lamivudine (DOL/3TC) demonstrated potent efficacy and favorable safety in both antiretroviral therapy-naïve and -experienced patients, but data on older people are lacking. We aimed to evaluate virological efficacy and safety of DOL/3TC in suppressed older patients over a 12-month period. We performed a retrospective cohort study evaluating people living with HIV (PLWHIV) aged ≥65 years at our HIV Clinic who were switched to DOL/3TC. Eligible patients had baseline HIV-1 RNA <20 copies/mL, and no previous virological failures or known resistance mutations for lamivudine or dolutegravir. Inclusion criteria were met by 72 patients: 59 were men, median age was 69.2 years, and one or more comorbidities were present in 89% of patients. The most common reason for switch was simplification, followed by drug-drug interactions (DDIs) and toxicities. After 12 months, 64 (88.9%, by the intention-to-treat analysis) patients maintained HIV-1 RNA <20 copies/mL, and reasons for treatment failure were virological failure in three cases, adverse events in three, and missing data in two. Genotype resistance testing showed no resistance mutations for lamivudine or dolutegravir in subjects with virological failure. The number of potential DDIs decreased from 92 to 12 after switching to DOL/3TC, and a significant reduction in median total and low-density lipoprotein cholesterol was reported, while median change in body weight was not significant. In this real-life cohort, switching to DOL/3TC was associated with maintenance of virological control and good tolerability among persons aged >65 years, supporting use of this dual regimen in older PLWHIV.


Potential of Tissue-Engineered and Artificial Dermis Grafts for Fingertip Reconstruction.
Management of skin and soft-tissue defects of the fingertips is functionally and aesthetically important, but controversial, especially when bones are exposed. Recent advances in wound healing technology allow the use of cells or biological dermis. The authors studied the clinical efficacy of tissue-engineered dermis grafts and artificial dermis grafts versus immediate reconstructive procedures, such as the reverse digital artery island flap, in treating bone-exposed fingertip defects. One hundred eighty-two patients with bone-exposed fingertip defects treated with tissue-engineered dermis grafts (n = 71), artificial dermis grafts (n = 23), or reverse digital artery island flaps (n = 88) were included in this retrospective cohort study. Surgical time, duration of hospitalization, total cost, success rate, healing time, sensory recovery, range of motion, scar quality, and patient satisfaction were compared. No tissue-engineered or artificial dermis graft exhibited graft rejection or failure, whereas there was one partial loss and one total loss after reverse digital artery island flap surgery. Tissue-engineered dermis grafts were superior in scar quality, and artificial dermis grafts had shorter surgical times and lower surgical costs; both groups demonstrated superior results in postoperative range of motion and sensory recovery in two-point discrimination tests and shorter hospitalization, compared with the reverse digital artery island flap group. The reverse digital artery island flap had shorter complete closure time and less postoperative tingling sensation. There were no differences in overall patient satisfaction among the groups. Tissue-engineered and artificial dermis grafts may be promising alternatives for fingertip reconstruction. In particular, tissue-engineered dermis grafts may deliver superior functional results, including recovery of sensory discomfort and aesthetic results in terms of scar quality over artificial dermis grafts. Therapeutic, III.


KSHV LANA1 binds DNA as an oligomer and residues N-terminal to the oligomerization domain are essential for DNA binding, replication, and episome persistence.
Latency-associated nuclear antigen 1 (LANA1) binds to Kaposi's sarcoma-associated herpesvirus (KSHV) terminal repeat (TR) DNA to mediate episome replication and persistence. LANA1 concentrates at sites of TR DNA along mitotic chromosomes, consistent with tethering KSHV DNA to chromosomes for efficient segregation of episomes to progeny nuclei. We now investigate LANA1 C-terminus self-association and DNA binding. The TR DNA binding domain was localized to LANA1 residues 996-1139. Scanning deletions within this region ablated both LANA1 oligomerization and DNA binding, consistent with a requirement for oligomerization to bind DNA. Furthermore, LANA1 bound TR DNA as an oligomer. Deletion of amino acids 1007-1021, N-terminal to the LANA1 oligomerization domain, ablated DNA binding, DNA replication, and episome persistence, implicating these residues in contacting DNA. Indeed, LANA1 residues 1007-1021 correspond to EBNA1 residues that contact the cognate sequence. Like EBNA1, the LANA1 DNA-binding domain has oligomerization activity and critical residues essential for recognizing DNA.


A simple and sensitive fluorescence biosensor for detection of organophosphorus pesticides using H2O2-sensitive quantum dots/bi-enzyme.
In this paper, we have developed a simple, fast, convenient and sensitive method for determination of organophosphorus pesticides in real samples based on inhibition mechanism of acetylcholinesterase (AChE). The biosensor is composed of enzymes (AChE and ChOx (choline oxidase)), QDs and acetylcholine (ACh), without any complex process of assembly for biosensor. After the experimental conditions are optimized, the limit of detection (LOD) for dichlorvos (DDVP) is found to be 4.49nM. Two linear ranges allow a wide determination of DDVP concentration from 4.49nM to 6780nM. Furthermore, a possible mechanism is put forward to explain the fluorescence quenching of CdTe QDs in the presence of H2O2. More importantly, the obtained biosensor is proven to be suitable for the detection of residues of organophosphorus pesticides (OPs) in real examples. The excellent performance of this biosensor will facilitate future development of rapid and high-throughput detection of organophosphorus pesticides.


Extracellular vesicles-encapsulated microRNA-10a-5p shed from cancer-associated fibroblast facilitates cervical squamous cell carcinoma cell angiogenesis and tumorigenicity via Hedgehog signaling pathway.
Cancer-associated fibroblast (CAF) secretes extracellular vesicle (EV)-encapsulated microRNAs (miRNAs) which have been underlined great promise for therapeutic target for diseases and cancers. Our study aimed to explore the role of EV-encapsulated miR-10a-5p from CAFs in angiogenesis in cervical cancer. Expression of miR-10a-5p in clinical samples of cervical cancer and cancer cells was detected by in situ hybridization and RT-qPCR. Results demonstrated that miR-10a-5p expression was upregulated in both cancer tissues and cells. CAFs and normal fibroblasts (NFs) from cervical cancer patient tissues were characterized under transmission electron microscopy, followed by EV isolation from CAFs. The EVs labeled with PKH67 were cultured with cervical squamous cell carcinoma (CSCC) cell line (SiHa) and HUVECs. Data indicated that CAF-EVs were internalized by cancer cells and promoted cell proliferation and tube formation. CAF-EVs then were transfected with miR-10a-5p inhibitor and then injected into nude mice. While injection of CAF-EVs promoted tumor growth and increased VEGFR and CD31 expression level, miR-10a-5p inhibitor-treated CAF-EVs resulted in decreased tumor volume and amount of vessel around tumor. Of note, dual-luciferase reporter gene assay and bioinformatic analysis indicated TBX5 as a target gene of miR-10a-5p. Moreover, EV-derived miR-10a-5p promoted angiogenesis in vivo and in vitro through activation of Hedgehog signaling via downregulation of TBX5. Taken altogether, miR-10a-5p in CAF-EVs promoted CSCC cell angiogenesis and tumorigenicity via activation of Hh signaling by inhibition of TBX5, providing insight into novel treatment based on miR-10a-5p against CSCC.


Hyperoxia sensitizes hypoxic HeLa cells to ionizing radiation by downregulating HIF‑1α and VEGF expression.
The current study investigated whether hyperoxia may reverse hypoxia‑induced radioresistance (RR) in cervical cancer. Human HeLa cells exposed to hypoxic, normoxic or hyperoxic conditions were irradiated using X‑rays. Cell proliferation and apoptosis were analyzed using MTT assays and flow cytometry. The expression levels of hypoxia‑inducible factor‑1α (HIF‑1α), VEGF165, VEGFRs, Akt and ERK were measured via western blotting and/or ELISA. The results demonstrated that hypoxia stimulated HIF‑1α and VEGF expression, and induced RR in HeLa cells. The administration of recombinant VEGF or the forced expression of VEGF promoted RR, whereas inactivating HIF‑1α or blocking the VEGF‑VEGFR interaction abrogated hypoxia‑induced RR. Notably, hyperoxia decreased the level of hypoxia‑stimulated HIF‑1α and VEGF, and enhanced radiosensitivity in hypoxic HeLa cells. The results demonstrated that hyperoxia suppressed the hypoxia‑activated Akt and ERK signaling pathways in HeLa cells. Therefore, a high O2 concentration may be considered as a radiotherapeutic sensitizer for hypoxic HeLa cells.


Comparative evaluation of the efficacy of 2% lidocaine containing 1:200,000 epinephrine with and without hyaluronidase (75 IU) in patients with irreversible pulpitis.
The purpose of this study was to determine the anesthetic efficacy of lidocaine containing epinephrine compared with lidocaine containing epinephrine plus hyaluronidase (75 IU) when performing an inferior alveolar nerve block. Patients complaining of pain in the mandibular posterior teeth were selected. Based on their chief complaint, proper clinical and radiographic examinations were performed. Among them, 40 subjects diagnosed with irreversible pulpitis were selected. The inferior alveolar nerve block was induced using 3 mL 2% lidocaine with epinephrine. Hyaluronidase (75 IU) or a placebo was injected 30 minutes after the beginning of pulpal anesthesia (randomized and double-blind trial). The duration of the effect in the pulpal and gingival tissues was evaluated by the response to painful electrical stimuli applied to the adjacent premolar and by mechanical stimuli (pinprick) to the buccal gingiva, respectively. In both pulpal and gingival tissues, the duration of the anesthetic effects with hyaluronidase was longer than with placebo. Hyaluronidase increased the duration of the effects of lidocaine in inferior alveolar nerve blocks.


Rapid estradiol-17beta modulation of opioid actions on the electrical and secretory activity of rat oxytocin neurons in vivo.
During pregnancy, emergence of endogenous opioid inhibition of oxytocin neurons is revealed by increased oxytocin secretion after administration of the opioid receptor antagonist, naloxone. Here we show that prolonged estradiol-17beta and progesterone treatment (mimicking pregnancy levels) potentiates naloxone-induced oxytocin secretion in urethane-anesthetized virgin female rats. We further show that estradiol-17beta alone rapidly modifies opioid interactions with oxytocin neurons, by recording their firing rate in anesthetized rats sensitized to naloxone by morphine dependence. Naloxone-induced morphine withdrawal strongly increased the firing rate of oxytocin neurons in morphine dependent rats. Estradiol-17beta did not alter basal oxytocin neuron firing rate over 30 min, but amplified naloxone-induced increases in firing rate. Firing pattern analysis indicated that acute estradiol-17beta increased oxytocin secretion in dependent rats by increasing action potential clustering without an overall increase in firing rate. Hence, rapid estradiol-17beta actions might underpin enhanced oxytocin neuron responses to naloxone in pregnancy.


New echocardiographic imaging method based on the bandwidth of the ultrasound Doppler signal with applications in blood/tissue segmentation in the left ventricle.
A new imaging method, Bandwidth Imaging, which is related to the bandwidth of the ultrasound Doppler signal is proposed as a classification function for blood and tissue signal in transthoracial echocardiography in the left ventricle. An in vivo experiment is presented, where the apparent error rate of Bandwidth Imaging is compared with the apparent error rate of Second-Harmonic Imaging on 15 healthy men. The apparent error rates are calculated from the 16 myocardial wall segments defined in [M.D., Cerqueira, N.J. Weissman, V. Dilsizian, A.K. Jacobs, S. Kaul, W.K. Laskey, D.J. Pennell, J.A. Rumberger, T. Ryan, M.S. Verani, Standardized myocardial segmentation and nomenclature for tomographic imaging of the heart, Circulation (2002) 539-542]. A hypothesis test of Bandwidth Imaging having lower apparent error rate than Second-Harmonic Imaging is proved for a p-value of 0.94 in three segments in end diastole and in one segment in end systole. When data was averaged by a structural element of five radial, three lateral and four temporal samples the numbers of segments increased to nine in end diastole and to six in end systole. This experiment indicates that Bandwidth Imaging can supply additional information for automatic border detection routines on endocardium.


A Randomized Controlled Crossover Study of the Impact of Online Music Training on Pitch and Timbre Perception in Cochlear Implant Users.
Cochlear implant (CI) biomechanical constraints result in impoverished spectral cues and poor frequency resolution, making it difficult for users to perceive pitch and timbre. There is emerging evidence that music training may improve CI-mediated music perception; however, much of the existing studies involve time-intensive and less readily accessible in-person music training paradigms, without rigorous experimental control paradigms. Online resources for auditory rehabilitation remain an untapped potential resource for CI users. Furthermore, establishing immediate value from an acute music training program may encourage CI users to adhere to post-implantation rehabilitation exercises. In this study, we evaluated the impact of an acute online music training program on pitch discrimination and timbre identification. Via a randomized controlled crossover study design, 20 CI users and 21 normal hearing (NH) adults were assigned to one of two arms. Arm-A underwent 1 month of online self-paced music training (intervention) followed by 1 month of audiobook listening (control). Arm-B underwent 1 month of audiobook listening followed by 1 month of music training. Pitch and timbre sensitivity scores were taken across three visits: (1) baseline, (2) after 1 month of intervention, and (3) after 1 month of control. We found that performance improved in pitch discrimination among CI users and NH listeners, with both online music training and audiobook listening. Music training, however, provided slightly greater benefit for instrument identification than audiobook listening. For both tasks, this improvement appears to be related to both fast stimulus learning as well as procedural learning. In conclusion, auditory training (with either acute participation in an online music training program or audiobook listening) may improve performance on untrained tasks of pitch discrimination and timbre identification. These findings demonstrate a potential role for music training in perceptual auditory appraisal of complex stimuli. Furthermore, this study highlights the importance and the need for more tightly controlled training studies in order to accurately evaluate the impact of rehabilitation training protocols on auditory processing.


Remote Ischemic Postconditioning (RIPC) After GMH in Rodents.
Germinal matrix hemorrhage (GMH) is the most common and devastating neurological injury of premature infants, and current treatment approaches are ineffective. Remote ischemic postconditioning (RIPC) is a method by which brief limb ischemic stimuli protect the injured brain. We hypothesized that RIPC can improve outcomes following GMH in rats. Neonatal rats (P7) were subjected to either stereotactic ganglionic eminence collagenase infusion or sham surgery. Groups were as follows: sham (n = 0), GMH non-RIPC (n = 10), GMH + 1 week RIPC (n = 10), GMH + 2 weeks RIPC (n = 10). Neurobehavior analysis at the fourth week consisted of Morris water maze (MWM) and rotarod (RR). This was followed by euthanasia for histopathology on day 28. Both 1- and 2-week RIPC showed significant improvement in FF and RR motor testing compared with untreated animals (i.e., GMH without RIPC). RIPC treatment also improved cognition (MWM) and attenuated neuropathological ventricular enlargement (hydrocephalus) in juvenile animals following GMH. RIPC is a safe and noninvasive approach that improved sensorimotor and neuropathological outcomes following GMH in rats. Further studies are needed to evaluate for mechanisms of neuroprotection.


The Department of Veterans' Affairs Depleted Uranium Cohort in the Time of COVID-19: Translating a Traditional Surveillance Protocol to a Telehealth Platform.
In 2021, 37 members of a cohort of depleted uranium-exposed Gulf War I veterans were evaluated using a protocol tailored to accommodate COVID-19 safety practices on a telehealth platform. Individual elements of the legacy protocol were reviewed for urgency and feasibility of inclusion in a modified, telehealth platform. The redesigned protocol included a participant readiness for telehealth assessment, nurse and physician telehealth visits, collection of usual health questionnaires, and urine collections for exposure monitoring for uranium and other fragment-related metal measures. Despite some limitations in scope, the telehealth platform permitted a visual "visit" with surveillance participants who expressed a high comfort level with the format. The telehealth platform has apparent utility for occupational surveillance and should be explored as a standard approach for surveillance outside of public health emergencies.


Effect of ageing on biochar properties and pollutant management.
Biochar is a carbon-rich pyrogenic material for multifunctional environmental applications such as carbon sequestration, soil amendment, and pollutant management, etc. Owing to long-term existing in the soil, biochar would inevitably suffer from natural geochemical weathering. Such ageing process could pose nonnegligible impacts on the physicochemical property and functionality of biochar. For an object-oriented design of biochar under different application ageing conditions, the latest research progress on ageing methods, biochar properties, and pollutant sorption performance needs to be fully understood. Specifically, the effect of soil components on biochar ageing is critically reviewed, which is of importance but not fully explored so far. The decrease of ash in aged biochar can inhibit the adsorption of heavy metals. The loss of aromatic components and the formation of three-dimensional water clusters during the ageing process have a negative impact on high-temperature biochar (>500° C) for organic pollutants adsorption. For long-term soil remediation, these results remind us to carefully use high-ash biochar for heavy metals and high-temperature biochar for organic pollutants. The interaction between soil minerals and biochar can form organometallic complexes and change functional groups to enhance the oxidation resistance of biochar. In the present review, the current research on biochar ageing are critical reviewed, and the further researches are prospected including developing advanced artificial ageing methods, exploring the impact of soil components on biochar ageing, and clarifying the long-term environmental behavior of modified biochar.


Survey of knowledge and attitude regarding induced abortion among nurses in a tertiary hospital in Thailand after amendment of the abortion act: a cross-sectional study.
The abortion act in Thailand is approximately 60 years old. However, because of increasing problems due to unsafe abortions, the act was recently amended to accord a legal status for abortions. In the southernmost provinces of Thailand, most people follow the Islamic faith, according to which induced abortion is a sin for both the providers and the pregnant women. This may affect the attitude of the medical staff, such as registered nurses, who play an important role in abortion services. Our study aims to evaluate the knowledge of the amended abortion act, attitude toward abortions and the intentions behind them, and willingness to perform abortions among registered nurses. A cross-sectional study was conducted from January 2022 to February 2022 wherein a self-administrated questionnaire was electronically distributed to 450 registered nurses practicing at a tertiary hospital in the southernmost province of Thailand. Linear regression analysis and Fisher's exact test were conducted to evaluate the association between basic characteristics, knowledge scores, and attitudes toward induced abortion. A total of 375 nurses (83.3%) completed the survey. Most participants were Muslim (58.9%), and 18.7% of them correctly answered > 80% of the knowledge questions. Among all the participants, 41.4% had a favorable attitude toward induced abortion, of which 21.3% were willing to provide safe abortion services. Knowledge scores were independently associated with practicing in obstetrics-gynecology departments and a lower age. Participants practicing Buddhism and having good knowledge scores tended to have favorable attitudes toward abortion. Nurses in the southernmost province of Thailand lack knowledge regarding the amended abortion act and do not have a favorable moral attitude toward abortion. Favorable attitudes toward abortions, support toward intentions behind abortions, and a willingness to provide abortion services were all lesser among the Muslim participants than among the Buddhist participants. Compared with participants who scored lower, those with higher knowledge scores had a better moral attitude toward abortion and, in turn, demonstrated a greater intention to provide abortion services. Encouraging nurses to gain better knowledge may improve their attitude toward abortion, which may positively influence future medical practices.


Anti-Helicobacter pylori activity of Fagopyrum Tataricum (L.) Gaertn. Bran flavonoids extract and its effect on Helicobacter pylori-induced inflammatory response.
Tartary buckwheat flavonoids have a variety of effects on anti-inflammatory, anti-oxidation, as well as anti-tumor and are valuable for academic research and industrial application. Helicobacter pylori (H. pylori) infection is associated with various gastrointestinal diseases in humans, and an increase in its resistance has led to the failure of many drugs. In this study, we quantified the main monomers of tartary buckwheat (Fagopyrum Tataricum (L.) Gaertn.) bran flavonoids extract through HPLC analysis. Then, we investigated the anti-H. pylori activity and the effect on cell inflammation of tartary buckwheat flavonoids extract and its four main flavonoid monomers (rutin, quercetin, kaempferol, and nicotiflorin). The results showed that tartary buckwheat flavonoids extract and its four flavonoid monomers could inhibit the growth of H. pylori and down-regulate the expression of proinflammatory factors IL-6, IL-8, and CXCL-1 in H. pylori-induced GES-1 cells. Moreover, we also confirmed that tartary buckwheat flavonoids extract could reduce the expression of virulence factor gene of H. pylori. In summary, tartary buckwheat can alleviate the cell inflammation induced by H. pylori, which provides a theoretical basis for the development of tartary buckwheat healthcare products.


Pregnancy outcomes in women with osteogenesis imperfecta.
To assess the relationship between osteogenesis imperfecta (OI) type, mode of delivery and outcomes as self-reported by women in the International Osteogenesis Imperfecta (OI) Registry. A cross-sectional study using data from 274 women with OI who reported their experience with pregnancy practices, including mode of delivery, number of children, genetic counseling, assisted conception techniques, mean ages at menarche and at menopause, and pregnancy complications. Chi-square analyses were performed to compare pregnancy outcomes, number of children and OI type. Prevalence ratios and 95% confidence intervals were obtained to quantify exposure-outcome relationships. The most common mode of delivery was the sole use of cesarean section (C-section) reported by 55% of the participants. Approximately two-thirds had at least two children. Twenty-nine percent (n = 80) reported pregnancy complications. There was a significant relationship between the mode of delivery and OI type (p < 0.001), genetic counseling (p = 0.010), and number of children (p < 0.0001). There was neither evidence of an association between pregnancy complications and number of children (p = 0.16), OI type (p = 0.27), nor mode of delivery (p = 0.11). These findings suggested that clinical OI type, pre-natal genetic counseling, and number of children were strong predictors for choosing the mode of delivery. Severity of OI, multiparity, and vaginal delivery were not associated with increased pregnancy complications in this cohort of women with OI.


Factors Influencing the Experience of Breast and Cervical Cancer Screening Among Women in Low- and Middle-Income Countries: A Systematic Review.
The persistent high morbidity and mortality from breast and cervical cancer in low- and middle-income countries (LMICs) may be influenced by global disparities in the uptake of screening services. This review sought to synthesize existing evidence to determine factors that influence the experience of women relating to breast and cervical screening in LMICs. A qualitative systematic review of the literature identified through Global Health, Embase, PsycInfo, and MEDLINE. Eligible studies included those outlining primary qualitative research or mixed-method studies with reporting of qualitative findings, detailing women's experiences of involvement with programs for breast or cervical cancer screening. Framework synthesis was used to explore and organize findings from primary qualitative studies and the Critical Appraisal Skills Programme checklist used for quality assessment. Database searches yielded 7,264 studies for title and abstract screening and 90 full-text articles for screening, with qualitative data from 17 studies and a total of 722 participants included in this review. Four stages influencing experiences of women were generated across both breast and cervical cancer screening approaches, with individual (eg, knowledge of cancer), social (eg, religion, cultural beliefs), and health system (eg, accessibility) factors identified that influence women's initial and subsequent engagement. This study synthesizes existing evidence of factors that influence engagement with breast and cervical cancer screening in LMICs. Evidence-informed recommendations are proposed that may improve the experience of cancer screening in LMICs, with further research necessary to explore their operationalization and impact on cancer care delivery.


Hypoxic and hyperoxic effect on blood phosphodiesterase activity in young and old rats.
In the present work we describe phosphodiesterase (PDEs) activity in the blood of young and old rats kept under hypoxic or hyperoxic normobaric conditions in order to correlate the age-change response with PDE levels and oxygen supply. PDE is important in the process of energy supply and as a modulator and mediator of several cellular functions. Three groups of Wistar rats were kept in room air, 10-12% oxygen for 12 days and 98-100% oxygen for 60 hrs respectively. Each group was composed of young rats (2 months of age) and old rats (25 months of age). After the exposure the rats were anaesthetized and blood samples were collected using an intracardiac catheter. The results show: a) in the control group, no significant difference between the PDE activities of old and young rats; b) a significant increase in PDE occurred after hypoxic and hyperoxic treatment in both young and aged rats; c) the increase in PDE activity was more evident in the young rather than the old rats; and d) the aged rats are less responsive to oxygen variation. The results demonstrate that young and aged rats respond to variations in the oxygen supply. Hypoxia and hyperoxia show different age-related intensity level response. We conclude that the alteration in PDE expression occurring in the blood as a consequence of hypoxic or hyperoxic treatment is probably a necessary protective response for the body against alteration from oxidative metabolism and to maintain the body in homeostatic ranges for energy requirements.


Bioassay for specific DNA sequences using a non-radioactive probe.
A novel method for detecting specific DNA sequences is described. The method uses a non-radioactive DNA probe, called a probe-vector, that can transform competent Escherichia coli cells at high efficiency only when it has hybridized to a specific DNA target, thus forming a circular, double-stranded, plasmid-like molecule. The probe-vector carries a plasmid origin of replication and a gene that confers antibiotic resistance on transformed E. coli. The output of the assay--colored bacterial colonies on an agar plate--is quantitative and proportional over a wide range of target concentrations. The utility of the probe-vector method for detecting hepatitis B virus (HBV) DNA in human serum is demonstrated. The assay can detect as little as 0.1 pg HBV DNA. The presence of an internal standard monitors DNA recovery and E. coli transformation efficiency for each sample. The assay has the potential to simultaneously measure the DNA of two or more pathogens within the same clinical sample.


Phase-preserving beam expander for biomedical X-ray imaging.
The BioMedical Imaging and Therapy beamlines at the Canadian Light Source are used by many researchers to capture phase-based imaging data. These experiments have so far been limited by the small vertical beam size, requiring vertical scanning of biological samples in order to image their full vertical extent. Previous work has been carried out to develop a bent Laue beam-expanding monochromator for use at these beamlines. However, the first attempts exhibited significant distortion in the diffraction plane, increasing the beam divergence and eliminating the usefulness of the monochromator for phase-related imaging techniques. Recent work has been carried out to more carefully match the polychromatic and geometric focal lengths in a so-called `magic condition' that preserves the divergence of the beam and enables full-field phase-based imaging techniques. The new experimental parameters, namely asymmetry and Bragg angles, were evaluated by analysing knife-edge and in-line phase images to determine the effect on beam divergence in both vertical and horizontal directions, using the flat Bragg double-crystal monochromator at the beamline as a baseline. The results show that by using the magic condition, the difference between the two monochromator types is less than 10% in the diffraction plane. Phase fringes visible in test images of a biological sample demonstrate that this difference is small enough to enable in-line phase imaging, despite operating at a sub-optimal energy for the wafer and asymmetry angle that was used.


Contraception for adolescents after abortion.
Preventing repeated unplanned pregnancy among adolescents is still a challenge because many of them fail to use effective contraception after abortion. To review currently recommended options of methods and counselling for effective prevention of repeat pregnancies in adolescents. Review of the literature that was identified through the Medline, ScienceDirect, Google and Popline databases and relevant expert opinions. Counselling needs to be adapted to the needs, values and lifestyle of adolescents. The best results are achieved with nondirective or active contraceptive counselling, followed by regular check-ups and cautious and attentive approach in the management of doubts, prejudices and side effects related to the contraceptive chosen. Adolescents should initiate contraception immediately after abortion: the motivation for choosing an efficacious method is highest at that time; resumption of ovulation following induced abortion occurs on average after three weeks; more than half of these girls will resume sexual activity within two weeks after pregnancy termination. Long-acting reversible contraception use during adolescence is safe and most effective. However, achieving a high long-term continuation rate is especially challenging in adolescents; this is due to developmental and environmental characteristics that influence their contraceptive behaviour. Adolescents should immediately after abortion initiate a reliable contraceptive method, preferably one whose efficacy is not user-dependent. Providing an appropriate health care would contribute to achieving continuity in the prevention of repeat pregnancy.


Gli1 is a potential cancer stem cell marker and predicts poor prognosis in ductal breast carcinoma.
Glioma-associated oncogene homolog 1 (Gli1) maintains the cancer stem cell-like characteristics in various tumors. However, its expression in cancer stem cells (CSC) in ductal breast carcinoma has not been well studied. We aimed to characterize Gli1 as a potential CSC marker and investigate its clinical significance in ductal breast carcinoma. Immunohistochemical staining was used to study the relationship of Gli1 to clinicopathologic features, cell cycle regulation-related genes, and CSC markers. Gli1 was expressed to a greater extent in ductal breast carcinoma than in normal breast tissues (P=.002). Its expression was significantly correlated with tumor grade (P=.044), pT stage (P=.017), and molecular subtype (P=.008). Expression was associated, not only with the expression of HIF-1α (P<.001), but also with greater microvessel density (MVD; P=.012). Kaplan-Meier survival analysis revealed that Gli1 was significantly associated with lower overall survival (OS; P=.02). Univariate Cox regression analysis confirmed that Gli1 was a poor prognostic factor for OS (P=.037) and was associated with the expression of the cell cycle-related genes cyclin D1 (P=.011), p21 (P=.009), and pAkt-Thr308 (P=.038). Moreover, Gli1 expression correlated significantly with the expression of two CSC markers, Sox2 (P=.01) and LSD1 (P=.01). Gli1 could be a stem cell marker and an indicator of poor prognosis in patients with ductal breast carcinoma.


Depoliticize human immunodeficiency virus infection: a commentary.
Public-health policy is inconsistent in its approach to the sexually transmitted disease human immunodeficiency virus (HIV). Nearly every health agency has politicized the reporting, finding, and contacting of HIV cases. There is also no consistency among the various state health departments and the various federal health agencies. Until we have a uniform health policy that treats HIV infection as every other reportable sexually transmitted disease, we will make little progress toward controlling its inevitable increase in both cases and costs.


Enzyme-linked immunosorbent assay for free thyroxin in human serum.
We describe a simple enzyme-linked immunosorbent assay (ELISA) in which microtiter plates are used for determining the free thyroxin concentration (FT4) in serum. Only commercially available chemicals and reagents are needed, including the antiserum. The working range is 1 to 60 ng/L. Turnaround time is about 4 h. Within-run coefficients of variation (CV) for FT4 concentrations of 9.0, 19.0, and 35.0 ng/L were less than 5%; between-run CVs were 10% to 11%. Results by a radioimmunoassay method for 150 sera correlated well (r = 0.922).


47,XY,+der(11;22)(q23;q12) following balanced translocation t(11;22)(q23;q12)mat. Remarks on the problem of trisomy 22.
Report of a supernumerary extra chromosome der(11;22)(q23;q12) resulting from a balanced translocation in the mother. The propositus suffers from mental deficiency, deafness and extreme muscular weakness and exhibits cleft palate, a labial lymphangioma and an atrial septum defect. Since the features of partial trisomy 11q23 frequently associated with a translocation t(11q;22q) bear similarities with the cases of so called trisomy 22 one might conjecture that some of these observations are in fact products of translocations including partial 11q.


BCG∆BCG1419c and BCG differ in induction of autophagy, c-di-GMP content, proteome, and progression of lung pathology in Mycobacterium tuberculosis HN878-infected male BALB/c mice.
The efficacy of BCG vaccines against Mycobacterium tuberculosis (Mtb) strains of lineage 2 (Beijing) in preclinical models and humans has been questioned. We have developed BCG∆BCG1419c, by deletion of BCG1419c in BCG Pasteur, which improved control of tuberculosis (TB) in preclinical models. Here, we compared the capacity of BCG and BCG∆BCG1419c to induce autophagy in murine macrophages, modify c-di-GMP content and transcript levels of BCG1416c, encoding the enzyme responsible for c-di-GMP synthesis/degradation, and of BCG1419c, encoding the phosphodiesterase involved in c-di-GMP degradation. Furthermore, we evaluated proteomic differences in vitro and compared protection against TB produced by a low dose of the HN878-Beijing strain at 3- and 6-months post-infection. We found that BCG∆BCG1419c induced more autophagy and produced different levels of c-di-GMP as well as different transcription of BCG1416c with no expression of BCG1419c. BCG∆BCG1419c differentially produced several proteins, including some involved in interaction with host cells. Vaccination with either BCG strain led to control of bacillary burden in lungs and spleen at 3- but not 6-months post-infection, whereas it reduced pneumonic areas compared with unvaccinated controls at 6 months post-infection. Vaccination with BCG∆BCG1419c delayed progression of lung necrosis as this was observed only at 6 months post-infection. Taken together, compared with BCG, BCG∆BCG1419c increased autophagy, presented different levels of c-di-GMP and transcription of BCG1416c in vitro in a growth-phase dependent manner, modified its proteome and delayed progression of lung pathology produced by a highly virulent Beijing strain.


Developmental changes in the human visual system as reflected by the latency of the pattern reversal VEP.
Pattern reversal visually evoked potentials (VEPs) were recorded from 439 infants and young children ranging in age from 1 month to 5 years in response to large and small checks. Qualitative analysis of the VEP wave form showed that the first major positive component, P1, is consistently present at all ages, while the frequency of occurrence of later positive components is more variable. The proportion of infants showing late positive components increases with age; by 1 year, the frequency of occurrence of late components for large checks is more adult-like than for small checks. The latency of P1 was analyzed quantitatively. Results showed that P1 latency decreases rapidly during the first year of life for both large and small checks and that the time course of the latency change differs as a function of check size. VEPs to large checks attain adult-like P1 latency values by about 1 year of age, while the P1 latency of VEPs to small checks has still not reached adult levels by 5 years of age. Data from 12 infants tested longitudinally between 1 and 7 months of age using both checkerboards and square wave gratings show no difference in P1 latency between checkerboards and gratings comprised of large (30-240 min) pattern elements, but for patterns with small (7.5 and 15 min) elements, P1 latency to checks is significantly longer than P1 latency to stripes. These results are explained on the basis of the difference in the fundamental spatial frequency between checks and stripes.


The therapeutic role of creatine in Huntington's disease.
Huntington's disease (HD) is an autosomal dominant and fatal neurological disorder characterized by a clinical triad of progressive choreiform movements, psychiatric symptoms, and cognitive decline. HD is caused by an expanded trinucleotide CAG repeat in the gene coding for the protein huntingtin. No proven treatment to prevent the onset or to delay the progression of HD currently exists. While a direct causative pathway from the gene mutation to the selective neostriatal neurodegeneration remains unclear, it has been hypothesized that interactions of the mutant huntingtin protein or its fragments may result in a number of interrelated pathogenic mechanisms triggering a cascade of molecular events that lead to the untimely neuronal death observed in HD. One putative pathological mechanism reported to play a prominent role in the pathogenesis of HD is mitochondrial dysfunction and the subsequent reduction of cellular energy. Indeed, if mitochondrial impairment and reduced energy stores play roles in the neuronal loss in HD, then a therapeutic strategy that buffers intracellular energy levels may ameliorate the neurodegenerative process. Sustained ATP levels may have both direct and indirect importance in ameliorating the severity of many of the pathogenic mechanisms associated with HD. Creatine, a guanidino compound produced endogenously and acquired exogenously through diet, is a critical component in maintaining much needed cellular energy. As such, creatine is one of a number of ergogens that may provide a relatively safe and immediately available therapeutic strategy to HD patients that may be the cornerstone of a combined treatment necessary to delay the relentless progression of HD.


Identifying evolutionary trees and substitution parameters for the general Markov model with invariable sites.
The general Markov plus invariable sites (GM+I) model of biological sequence evolution is a two-class model in which an unknown proportion of sites are not allowed to change, while the remainder undergo substitutions according to a Markov process on a tree. For statistical use it is important to know if the model is identifiable; can both the tree topology and the numerical parameters be determined from a joint distribution describing sequences only at the leaves of the tree? We establish that for generic parameters both the tree and all numerical parameter values can be recovered, up to clearly understood issues of 'label swapping'. The method of analysis is algebraic, using phylogenetic invariants to study the variety defined by the model. Simple rational formulas, expressed in terms of determinantal ratios, are found for recovering numerical parameters describing the invariable sites.


Bacteria isolated from deep joint sepsis after operation for total hip or knee replacement and the sources of the infections with Staphylococcus aureus.
A wide variety of bacterial species, many usually regarded as of low pathogenicity, were isolated from septic joints after operation for total hip or knee joint replacement in a multi-centre trial of ultraclean air in operating rooms. The prophylactic antibiotics generally used appeared to reduce considerably the rates of infection with most species but to be ineffective against 'gut' organisms. For about half the septic infections involving Staphylococcus aureus nasal swabs had been obtained from the patient and operating staff at the time of operation for insertion of the prosthesis. Strains of Staph, aureus isolated from these and from the infected joints were phage typed and tested for antibiotic sensitivity. A probable source among the carriers was found for seven out of the 14 infections and a possible source for another five. Very small numbers of Staph. aureus were needed to initiate infection.


Ultradeformable Lipid Vesicles Localize Amphotericin B in the Dermis for the Treatment of Infectious Skin Diseases.
Cutaneous fungal and parasitic diseases remain challenging to treat, as available therapies are unable to permeate the skin barrier. Thus, treatment options rely on systemic therapy, which fail to produce high local drug concentrations but can lead to significant systemic toxicity. Amphotericin B (AmB) is highly efficacious in the treatment of both fungal and parasitic diseases such as cutaneous leishmaniasis but is reserved for parenteral administration in patients with severe pathophysiology. Here, we have designed and optimized AmB-transfersomes [93.5% encapsulation efficiency, 150 nm size, and good colloidal stability (-35.02 mV)] that can remain physicochemically stable (>90% drug content) at room temperature and 4 °C over 6 months when lyophilized and stored under desiccated conditions. AmB-transfersomes possessed good permeability across mouse skin (4.91 ± 0.41 μg/cm2/h) and 10-fold higher permeability across synthetic Strat-M membranes. In vivo studies after a single topical application in mice showed permeability and accumulation within the dermis (>25 μg AmB/g skin 6 h postadministration), indicating the delivery of therapeutic amounts of AmB for mycoses and cutaneous leishmaniasis, while a single daily administration in Leishmania (Leishmania) amazonensis infected mice over 10 days, resulted in excellent efficacy (98% reduction in Leishmania parasites). Combining the application of AmB-transfersomes with metallic microneedles in vivo increased the levels in the SC and dermis but was unlikely to elicit transdermal levels. In conclusion, AmB-transfersomes are promising and stable topical nanomedicines that can be readily translated for parasitic and fungal infectious diseases.


The role of peer physical activity champions in the workplace: a qualitative study.
Peer health champions have been suggested as an important component of multilevel workplace interventions to promote healthy behaviours such as physical activity (PA). There is accumulating quantitative evidence of their effectiveness but as yet little exploration of why and how champions influence peer behaviour. The current study explores the role of peer physical activity champions (PPACs) in influencing colleagues' PA behaviour from the perspectives of both champions and colleagues. Seven months after the introduction of a workplace PA programme in 17 small- and medium-sized enterprises (SMEs), two focus groups were held with PPACs and four with programme participants. Data were analysed using inductive thematic analysis. Three overarching themes were developed: how PPACs encourage PA, valuable PPAC characteristics and sustaining motivation for the PPAC role. Both direct encouragement from PPACs and facilitation of wider PA supportive social networks within the workplace encouraged behaviour change. PA behaviour change is a delicate subject and it was important that PPACs provided enthusiastic and persistent encouragement without seeming judgemental. Being a PA role model was also a valuable characteristic. The PPACs found it satisfying to see positive changes in their colleagues who had become more active. However, colleagues often did not engage in suggested activities and PPACs required resilience to maintain personal motivation for the role despite this. Incorporating PPACs into SME-based PA interventions is acceptable to employees. It is recommended that PPAC training includes suggestions for facilitating social connections between colleagues. Sensitivity is required when initiating and engaging in conversations with colleagues about increasing their PA. Programmes should ensure PPACs themselves are provided with social support, especially from others in the same role, to help sustain motivation for their role. These findings will be useful to health-promotion professionals developing workplace health programmes.


Innate and adaptive immune responses to the major Parietaria allergen Par j 1 in healthy subjects.
In this study we wanted to analyse the pattern of the immune response to the Parietaria major allergen Par j 1 in freshly purified peripheral blood mononuclear cell (PBMC) from healthy subjects. We observed that Par j 1 was capable of inducing IFN-γ production by CD3⁻ and CD16⁺/CD56⁺ cells exclusively in healthy individuals. Furthermore, a multiparametric analysis allowed us a better definition of two IFN-γ-Par j 1 specific populations (IFN-γ(dim) and IFN-γ(high)) characterized by the presence of different proportions of NKT and NK cells. We also identified the concomitant presence of a subset of IL-10⁺ NK cells. Moreover, CFSE staining showed that the Par j 1 preferentially induced the proliferation of CD3⁻/CD56⁺/CD335⁺ cells. Finally, a subset of CD4⁺/CD25⁺/FoxP3⁺/IL-10⁻ T cells was identified. The result of this pilot study suggest that during a tolerogenic response, the major allergen of the Parietaria pollen works as an activator of both the innate and the adaptive human immune system.


Applications of an inactive Dispersin B probe to monitor biofilm polysaccharide production.
Bacterial biofilms consist of surface-attached communities that secrete polymeric substances to form a biofilm matrix, generating a local microenvironment which helps protect from external factors. One such matrix component produced by a diverse list of microorganisms is the polysaccharide poly-β-1,6-N-acetylglucosamine (PNAG). Dispersin B is a PNAG-specific glycosyl hydrolase, which by leveraging its unique specificity, can be used to design a macromolecular fluorescent PNAG binding probe. An active site mutant of Dispersin B was fused to a fluorescent protein, to generate a probe that bound PNAG but did not hydrolyze its polysaccharide target. The ease and versatility of this strategy has made it possible to study PNAG in the context of maturing biofilms, as the probe tends to sequester in regions of high PNAG density. In this chapter, typical workflows from probe construction to cell-binding and imaging experiments are described.


A Randomized Phase 2 Trial of Antibiotic Prophylaxis Versus No Intervention for Muscle Biopsy in A Neurology Department.
Muscle biopsy can be used to confirm the diagnosis of neuromuscular diseases. However, it is unclear whether antibiotic prophylaxis prior to muscle biopsy is needed to prevent surgical site infection (SSI). We are conducting a phase 2, single-center, open-labeled, prospective randomized trial to clarify the need for antibiotic prophylaxis in patients at low risk for SSI undergoing muscle biopsy. Patients will be randomized to an antibiotic prophylaxis group or a control group, and the incidence of SSI will be compared between the groups. Our findings will clarify the need for antibiotic prophylaxis in this patient population.


Performance of concurrent tasks: a psychophysiological analysis of the reciprocity of information-processing resources.
The resources allocated to a primary and secondary task are reciprocal. Subjects performed a tracking task in which the discrete displacements of the tracking cursor could be used to elicit event-related brain potentials. As the resource demands of the tracking task were increased, potentials elicited by the task-defined events increased in amplitude, whereas those elicited by secondary task auditory stimuli decreased.


Cyclic nucleotide regulation of PAI-1 mRNA stability. Identification of cytosolic proteins that interact with an a-rich sequence.
Incubation of HTC rat hepatoma cells with the cyclic nucleotide analogue 8-bromo-cAMP results in a 3-fold increase in the rate of degradation of type-1 plasminogen activator-inhibitor (PAI-1) mRNA. Previous studies utilizing HTC cells stably transfected with beta-globin:PAI-1 chimeric constructs demonstrated that at least two regions within the PAI-1 3'-untranslated region mediate the cyclic nucleotide-induced destabilization of PAI-1 mRNA; one of these regions is the 3'-most 134 nucleotides (nt) of the PAI-1 mRNA (Heaton, J. H., Tillmann-Bogush, M., Leff, N. S., and Gelehrter, T. D. (1998) J. Biol. Chem. 273, 14261-14268). In the present study, ultraviolet cross-linking analyses of this region demonstrate HTC cell cytosolic mRNA-binding proteins ranging from 38 to 76 kDa, with a major complex migrating at approximately 50 kDa. RNA electrophoretic mobility shift analyses demonstrate high molecular weight multiprotein complexes that specifically interact with the 134-nt cyclic nucleotide-responsive sequence. The 50, 61, and 76 kDa and multiprotein complexes form with an A-rich sequence at the 3' end of the cyclic nucleotide-responsive region; a 38-kDa complex forms with a U-rich region at the 5' end of the 134 nt sequence. Mutation of the A-rich region prevents both the binding of the 50-, 61-, and 76-kDa proteins and formation of the multiprotein complexes, as well as cyclic nucleotide-regulated degradation of chimeric globin:PAI-1 transcripts in HTC cells. These data suggest that the proteins identified in this report play an important role in the cyclic nucleotide regulation of PAI-1 mRNA stability.


Assessing clinicopathological correlation in chronic traumatic encephalopathy: rationale and methods for the UNITE study.
Chronic traumatic encephalopathy (CTE) is a progressive neurodegeneration associated with repetitive head impacts. Understanding Neurologic Injury and Traumatic Encephalopathy (UNITE) is a U01 project recently funded by the National Institute of Neurological Disorders and Stroke and the National Institute of Biomedical Imaging and Bioengineering. The goal of the UNITE project is to examine the neuropathology and clinical presentation of brain donors designated as "at risk" for the development of CTE based on prior athletic or military exposure. Here, we present the rationale and methodology for UNITE. Over the course of 4 years, we will analyze the brains and spinal cords of 300 deceased subjects who had a history of repetitive head impacts sustained during participation in contact sports at the professional or collegiate level or during military service. Clinical data are collected through medical record review and retrospective structured and unstructured family interviews conducted by a behavioral neurologist or neuropsychologist. Blinded to the clinical data, a neuropathologist conducts a comprehensive assessment for neurodegenerative disease, including CTE, using published criteria. At a clinicopathological conference, a panel of physicians and neuropsychologists, blinded to the neuropathological data, reaches a clinical consensus diagnosis using published criteria, including proposed clinical research criteria for CTE. We will investigate the validity of these clinical criteria and sources of error by using recently validated neuropathological criteria as a gold standard for CTE diagnosis. We also will use statistical modeling to identify diagnostic features that best predict CTE pathology. The UNITE study is a novel and methodologically rigorous means of assessing clinicopathological correlation in CTE. Our findings will be critical for developing future iterations of CTE clinical diagnostic criteria.


Shear wave elastography in varicocele patients: Prospective study to investigate correlation with semen parameters and histological findings.
Shear wave elastosonography (SWE) is a non-invasive ultrasound imaging modality used to assess the mechanical properties of tissues such as rigidity and elasticity. In this prospective study, we investigated the effect of laparoscopic varicocelectomy on the elasticity, degree of fibrosis and function of the testes through SWE and we evaluated the correlation with semen parameters and histology findings. Male patients with monolateral left varicocele and progressive alteration of the semen quality were enrolled prospectively. Patients were evaluated before varicocelectomy, 3 and 6 months after surgery with semen analysis, ecocolordoppler US and SWE. In all patients, a left testicular biopsy was performed at the time of varicocelectomy and it was repeated after 6 months in 55% of patients in order to investigate the histological findings and to correlate with SWE results. The study was conducted on 82 patients. SWE showed a statistically significant difference between left and right testicles. Three months after surgery the mean left testicular volume increased, mean left SWE features decreased, and sperm count increased (P values < .0001). The SWE parameters, testicular volume and semen analysis values showed a statistically significant positive correlation between the pre and postoperative results (P value < .0001). The histological alterations were significantly changed 6 months postoperative with a complete morphology recovery in accordance with SWE results. SWE showed a statistically significant positive correlation with testicular volume, semen analysis and histological findings. This study represents the first investigation that correlates the varicocele, the testis volume, the quality of the seminal fluid ant the histological findings with the ultrasound and SWE values.


Comparison of Rates of Type 2 Diabetes in Adults and Children Treated With Anticonvulsant Mood Stabilizers.
Anticonvulsant mood stabilizer treatment is associated with an increased risk of weight gain, but little is known about the risk of developing type 2 diabetes (T2D). To evaluate the comparative safety of anticonvulsant mood stabilizers on risk of T2D in adults and children by emulating a target trial. This observational cohort study used data from IBM MarketScan (2010-2019), with a 5-year follow-up period. The nationwide sample of US commercially insured patients included children (aged 10-19 years) and adults (aged 20-65 years) who initiated anticonvulsant mood stabilizer treatment. Data were analyzed from August 2020 to May 2021. Initiation and continuation of carbamazepine, lamotrigine, oxcarbazepine, or valproate. Onset of T2D during follow-up. Weighted pooled logistic regression was used to estimate the association of initiation and continuation of carbamazepine, lamotrigine, oxcarbazepine, or valproate with the risk of developing T2D. Inverse probability weights were used to control for confounding and loss to follow-up by measured baseline and time-varying covariates. The analysis included 274 206 adults (159 428 women [58%]; mean [SD] age, 39.9 [13.2] years) and 74 005 children (38 672 girls [52%]; mean [SD] age, 15.6 [2.6] years) who initiated an anticonvulsant mood stabilizer. In adults, initiation of valproate was associated with an increased risk of developing T2D compared with initiation of lamotrigine (5-year risk difference [RD], 1.17%; 95% CI, 0.66% to 1.76%). The number needed to harm was 87 patients initiating valproate for 1 patient to develop T2D within 5 years compared with initiation of lamotrigine. Point estimates were similar when evaluating the association of treatment continuation (5-year RD, 1.99%; 95% CI, -0.64% to 5.31%). The estimated association was smaller and more variable comparing carbamazepine and oxcarbazepine to lamotrigine. In children, RDs were much smaller and more variable (5-year RD for initiation of oxcarbazepine vs lamotrigine, 0.29%; 95% CI, -0.12% to 0.69%; 5-year RD for initiation of valproate vs lamotrigine, 0.18%; 95% CI, -0.09% to 0.49%). In this cohort study, valproate was associated with the highest risk of developing T2D in adults. The comparative safety was generally similar in children, but estimates were small and variable. In the absence of randomized trials, emulating target trials within health care databases can generate the age-specific drug safety data needed to inform treatment decision-making.


Crocein Orange G mediated detection and modulation of amyloid fibrillation revealed by surface-enhanced Raman spectroscopy.
Protein fibrous aggregation is associated with many neurodegenerative diseases including Alzheimer's and Parkinson's diseases. To modulate the process, a number of fibrillation inhibitors have been reported, although their working mechanism remains vague, calling for new means to decipher their interaction. Herein, we identified and characterized a novel inhibitor called Crocein Orange G (COG), which inhibited the nucleation and impeded the protofibril formation, revealed by various experimental approaches as well as molecular docking. In particular, the surface-enhanced Raman spectroscopy (SERS) helps to identify the binding sites and illustrate the interaction mechanism and fibrillation process by using Ag IMNPs as SERS substrate for a label-free detection. Combining with molecular docking, the SERS-based approach provides structural information concerning protein-ligand interaction and protein fibrillation. This study suggests that SERS can be a powerful new means to study the interaction between inhibitors and amyloid proteins and can potentially be a common tool for amyloid research. Strikingly, the SERS signal of COG corresponds very well with the state of protein fibrillation, hinting its function as an amyloid SERS signal amplifier. Therefore, this study provides a new means to monitor and interfere amyloid fibrillation.


Treatment with suboptimal doses of raltegravir leads to aberrant HIV-1 integrations.
Integration of the DNA copy of the HIV-1 genome into a host chromosome is required for viral replication and is thus an important target for antiviral therapy. The HIV-encoded enzyme integrase (IN) catalyzes two essential steps: 3' processing of the viral DNA ends, followed by the strand transfer reaction, which inserts the viral DNA into host DNA. Raltegravir binds to IN and blocks the integration of the viral DNA. Using the Rous sarcoma virus-derived vector RCAS, we previously showed that mutations that cause one viral DNA end to be defective for IN-mediated integration led to abnormal integrations in which the provirus had one normal and one aberrant end, accompanied by rearrangements in the host genome. On the basis of these results, we expected that suboptimal concentrations of IN inhibitors, which could block one of the ends of viral integration, would lead to similar aberrant integrations. In contrast to the proviruses from untreated cells, which were all normal, ∼10-15% of the proviruses isolated after treatment with a suboptimal dose of raltegravir were aberrant. The aberrant integrations were similar to those seen in the RCAS experiments. Most of the aberrant proviruses had one normal end and one aberrant end and were accompanied by significant rearrangements in the host genome, including duplications, inversions, deletions and, occasionally, acquisition of sequences from other chromosomes. The rearrangements of the host DNA raise concerns that these aberrant integrations might have unintended consequences in HIV-1-infected patients who are not consistent in following a raltegravir-containing treatment regimen.