Blood Pressure Regulation by the Carotid Sinus Nerve: Clinical Implications for Carotid Body Neuromodulation.
Chronic carotid sinus nerve (CSN) electrical modulation through kilohertz frequency alternating current improves metabolic control in rat models of type 2 diabetes, underpinning the potential of bioelectronic modulation of the CSN as a therapeutic modality for metabolic diseases in humans. The CSN carries sensory information from the carotid bodies, peripheral chemoreceptor organs that respond to changes in blood biochemical modifications such as hypoxia, hypercapnia, acidosis, and hyperinsulinemia. In addition, the CSN also delivers information from carotid sinus baroreceptors-mechanoreceptor sensory neurons directly involved in the control of blood pressure-to the central nervous system. The interaction between these powerful reflex systems-chemoreflex and baroreflex-whose sensory receptors are in anatomical proximity, may be regarded as a drawback to the development of selective bioelectronic tools to modulate the CSN. Herein we aimed to disclose CSN influence on cardiovascular regulation, particularly under hypoxic conditions, and we tested the hypothesis that neuromodulation of the CSN, either by electrical stimuli or surgical means, does not significantly impact blood pressure. Experiments were performed in Wistar rats aged 10-12 weeks. No significant effects of acute hypoxia were observed in systolic or diastolic blood pressure or heart rate although there was a significant activation of the cardiac sympathetic nervous system. We conclude that chemoreceptor activation by hypoxia leads to an expected increase in sympathetic activity accompanied by compensatory regional mechanisms that assure blood flow to regional beds and maintenance of hemodynamic homeostasis. Upon surgical denervation or electrical block of the CSN, the increase in cardiac sympathetic nervous system activity in response to hypoxia was lost, and there were no significant changes in blood pressure in comparison to control animals. We conclude that the responses to hypoxia and vasomotor control short-term regulation of blood pressure are dissociated in terms of hypoxic response but integrated to generate an effector response to a given change in arterial pressure.


A matrix metalloproteinase mediates long-distance attenuation of stem cell proliferation.
Ligand-based signaling can potentiate communication between neighboring cells and between cells separated by large distances. In the Drosophila melanogaster ovary, Wingless (Wg) promotes proliferation of follicle stem cells located ~50 µm or five cell diameters away from the Wg source. How Wg traverses this distance is unclear. We find that this long-range signaling requires Division abnormally delayed (Dally)-like (Dlp), a glypican known to extend the range of Wg ligand in the wing disc by binding Wg. Dlp-mediated spreading of Wg to follicle stem cells is opposed by the extracellular protease Mmp2, which cleaved Dlp in cell culture, triggering its relocalization such that Dlp no longer contacted Wg protein. Mmp2-deficient ovaries displayed increased Wg distribution, activity, and stem cell proliferation. Mmp2 protein is expressed in the same cells that produce Wg; thus, niche cells produce both a long-range stem cell proliferation factor and a negative regulator of its spreading. This system could allow for spatial control of Wg signaling to targets at different distances from the source.


Universal Health Coverage in Marginalized Populations: A Qualitative Evaluation of a Health Reform Implementation in Rural Ecuador.
In 2008, Ecuador underwent a major health reform with the aim of universal coverage. Little is known about the implementation of the reform and its perceived effects in rural parts of the country. The aim of this study was to explore the perceived effects of the 2008 health reform implementation, on rural primary health care services and financial access of the rural poor. A qualitative study using focus group discussions was conducted in a rural region in Ecuador, involving health staff, local health committee members, village leaders, and community health workers. Qualitative content analysis focusing on the manifest content was applied. Three categories emerged from the texts: (1) the prereform situation, which was described as difficult in terms of financial access and quality of care; (2) the reform process, which was perceived as top-down and lacking in communication by the involved actors; lack of interest among the population was reported; (3) the effects of the reform, which were mainly perceived as positive. However, testimonies about understaffing, drug shortages, and access problems for those living furthest away from the health units show that the reform has not fully achieved its intended effects. New problems are a challenging health information system and people without genuine care needs overusing the health services. The results indicate that the Ecuadorean reform has improved rural primary health care services. Still, the reform faces challenges that need continued attention to secure its current achievements and advance the health system further.


[Short- and long-term potentiation in the afferent pathways of the hippocampus in the newborn rabbit].
Field potentials were recorded from dendritic and cell body layers of CA1 and dentate regions in newborn rabbits. Responses were evoked by electrical stimulation of the collateral Schaffer system and the perforant path with paired (a 15-100 ms interstimulus interval) and rhythmical stimuli (20-40/c, 3-5 s). Short-term potentiation in the tests with paired stimulation and posttetanic potentiation (up to 3 h) were observed in the CA1 area beginning from the first postnatal day and in the later maturing fascia dentata beginning from 8-10th days, i.e. when neuronal reactions to stimulation of the appropriate afferent pathways first appeared.


Common menstrual problems in adolescence.
Many girls tell their general practitioners and paediatricians about problems with their periods. This article focuses on a practical approach to managing menorrhagia, dysmenorrhoea, oligomenorrhoea and premenstrual syndrome (PMS) and reviews the literature on this topic.


Threshold driven contagion on weighted networks.
Weighted networks capture the structure of complex systems where interaction strength is meaningful. This information is essential to a large number of processes, such as threshold dynamics, where link weights reflect the amount of influence that neighbours have in determining a node's behaviour. Despite describing numerous cascading phenomena, such as neural firing or social contagion, the modelling of threshold dynamics on weighted networks has been largely overlooked. We fill this gap by studying a dynamical threshold model over synthetic and real weighted networks with numerical and analytical tools. We show that the time of cascade emergence depends non-monotonously on weight heterogeneities, which accelerate or decelerate the dynamics, and lead to non-trivial parameter spaces for various networks and weight distributions. Our methodology applies to arbitrary binary state processes and link properties, and may prove instrumental in understanding the role of edge heterogeneities in various natural and social phenomena.


The prevalence of selected chronic diseases among the Mexican-American elderly: data from the 1982-1984 Hispanic Health and Nutrition Examination Survey.
Although the Mexican-American elderly represent a large and rapidly growing subgroup of the ethnic aged, national prevalence data of major chronic diseases among this population are sparse. Data based on physical examinations from three older groups of the southwestern portion of the 1982-1984 Hispanic Health and Nutrition Examination Survey (HHANES) were reviewed to determine rates of hypertension, diabetes mellitus, arthritis, and heart disease. The results show a higher prevalence of diabetes and lower prevalence of heart disease and hypertension when compared with the general population, coinciding with data from previous studies. Findings for the prevalence of arthritis varied widely from other studies, however, and proportions were lower than expected. Further research on arthritis in the Mexican-American elderly is necessary to determine whether protective mechanisms that may lead to a lower prevalence of this disease exist in this ethnic subgroup.


A comprehensive assessment of N-terminal signal peptides prediction methods.
Amino-terminal signal peptides (SPs) are short regions that guide the targeting of secretory proteins to the correct subcellular compartments in the cell. They are cleaved off upon the passenger protein reaching its destination. The explosive growth in sequencing technologies has led to the deposition of vast numbers of protein sequences necessitating rapid functional annotation techniques, with subcellular localization being a key feature. Of the myriad software prediction tools developed to automate the task of assigning the SP cleavage site of these new sequences, we review here, the performance and reliability of commonly used SP prediction tools. The available signal peptide data has been manually curated and organized into three datasets representing eukaryotes, Gram-positive and Gram-negative bacteria. These datasets are used to evaluate thirteen prediction tools that are publicly available. SignalP (both the HMM and ANN versions) maintains consistency and achieves the best overall accuracy in all three benchmarking experiments, ranging from 0.872 to 0.914 although other prediction tools are narrowing the performance gap. The majority of the tools evaluated in this study encounter no difficulty in discriminating between secretory and non-secretory proteins. The challenge clearly remains with pinpointing the correct SP cleavage site. The composite scoring schemes employed by SignalP may help to explain its accuracy. Prediction task is divided into a number of separate steps, thus allowing each score to tackle a particular aspect of the prediction.


Does Pneumococcal Vaccination Have an Effect on Hospital Costs?
It is known that inpatient hospital costs are much higher than outpatient services. It was aimed to investigate the effect of pneumococcal vaccination on hospitalizations. The direct hospitalization costs, length of stay, and factors of the vaccinated and unvaccinated patients in the same hospital during the 12-month follow-up of the patients who received pneumococcal vaccine between November 15, 2018, and November 15, 2020, in 3 chest diseases and thoracic surgery training and research hospitals were analyzed by obtaining Hospital Information Management System records. Data were collected with Statistical Package for the Social Sciences version 23 program (IBM Corp.; Armonk, NY, USA) , and statistical evaluation was made. The mean age of 800 hospitalized patients, of whom 400 were unvaccinated and 400 were vaccinated, was 68.48 ± 11.97. There was no significant difference in the mean age of vaccinated and unvaccinated patients (P > .05). Five hundred sixty-six patients (70.8%) were aged 65 and over. Two hundred eighty (51.2%) of men were vaccinated and 120 (47.2%) of women were vaccinated, and there was no significant difference (P > .05). The mean hospital stay of these patients was 11.01 days, and those in the vaccinated group had an average mean hospital stay of 9.11 days and those in the unvaccinated group had a mean hospital stay less than 12.91 days (P < .001). Total 1-year hospitalization costs were $501.653.53 and the cost per person was calculated as $627.07. The cost per capita for the vaccinated group was $550.52, which was lower than the average cost of the unvaccinated group ($703.62) (P < .05). When comparing the status of being vaccinated, comorbidity, mortality, mean length of stay, chronic obstructive pulmonary disease, and heart disease were found to be statistically significant (P < .05). In our study, it was revealed that vaccination of patients hospitalized in chest disease hospitals with the pneumococcal vaccine reduced the average length of hospital stay by 41.7% and the cost of hospitalization by 27.8%.


Simvastatin modulates angiotensin II signaling pathway by preventing Rac1-mediated upregulation of p27.
Recent experimental observations have suggested that statins may exert modulatory effects on a number of pathobiological processes beyond their cholesterol-lowering properties. Some of the pleiotropic effects of statins seem to be mediated by their ability to block the synthesis of isoprenoid intermediates, which serve as important lipid attachments required for the proper function and activation of the small GTP-binding proteins. The current study explored the modulatory effects of simvastatin (SMV) on the angiotensin II (Ang II)-induced Rac1-mediated, upregulation of cyclin-dependent kinase inhibitor p27. Ang II (100 nM) stimulation of rat mesangial cells induced a significant increase in p27 protein expression. Co-treatment of cells with SMV (1 microM) inhibited Ang II-induced upregulation of p27 protein. Addition of mevalonate (200 microM) or geranylgeranyl pyrophosphate (5 microM) reversed the inhibitory effect of SMV on p27 protein expression, suggesting that the effect of SMV is geranylgeranyl dependent. This study also provides evidence for a sequential link between Ang II stimulation and downstream activation of Rac1, intracellular H2O2 production, and Akt kinase leading to upregulation of p27 protein in mesangial cells. It was also shown that SMV, by inhibiting Rac1 activity, reversed Ang II-induced increase in intracellular H2O2 production, Akt activation, and p27 protein expression. The data presented in this study not only elucidate Ang II-mediated signaling cascade in mesangial cells but also demonstrate for the first time the modulatory effects of SMV on Ang II-induced signaling pathway at the cell cycle level.


Actinorhizal plant defence-related genes in response to symbiotic Frankia.
Actinorhizal plants have become increasingly important as climate changes threaten to remake the global landscape over the next decades. These plants are able to grow in nutrient-poor and disturbed soils, and are important elements in plant communities worldwide. Besides that, most actinorhizal plants are capable of high rates of nitrogen fixation due to their capacity to establish root nodule symbiosis with N2-fixing Frankia strains. Nodulation is a developmental process that requires a sequence of highly coordinated events. One of these mechanisms is the induction of defence-related events, whose precise role in a symbiotic interaction remains to be elucidated. This review summarises what is known about the induction of actinorhizal defence-related genes in response to symbiotic Frankia and their putative function during symbiosis.


Identification of character impact odorants of different soybean lecithins.
The potent odorants of standardized, enzymatically hydrolyzed, and deoiled soybean lecithins were characterized systematically by combined gas chromatography/mass spectrometry and olfactometry. Sixty-one odorants were identified; 53 of these odor-active compounds have not previously been reported as odorants of soybean lecithin flavor. By aroma extract dilution analysis and modified combined hedonic and response measurement the following odorants showed the highest flavor dilution factors and CHARM values: (E,E)-2, 4-decadienal (deep-fried), (E)-beta-damascenone (apple-like), 2, 3-diethyl-5-methylpyrazine (roasty, earthy), (E)-2-nonenal (cardboard-like), trans-4,5-epoxy-(E)-2-decenal (metallic), 1-nonen-3-one (mushroom-like), 2-ethyl-3,5-dimethylpyrazine (roasty, earthy), and 1-octen-3-one (mushroom-like). Enzymatic hydrolysis intensified especially the roasty sensation of 2, 3-diethyl-5-methylpyrazine, whereas deoiling effected a general significant decrease in olfactory perception on the nitrogen-containing compounds. In addition, sensory profiles of nasal and retronasal lecithin odor were performed.


Vitamin K-dependent proteins in the developing and aging nervous system.
Although the warfarin embryopathy syndrome, with its neurologic and bone abnormalities, has been known for decades, the role of vitamin K in the brain has not been studied systematically. Recently, it was demonstrated that vitamin K-dependent carboxylase expression is temporally regulated in a tissue-specific manner with high expression in the nervous system during the early embryonic stages and with liver expression after birth and in adult animals. This finding, along with the discovery of wide distribution of the novel vitamin K-dependent growth factor, Gas6, in the central nervous system, provides compelling evidence of a biologic role of vitamin K during the development of the nervous system. In animals and bacteria, vitamin K was observed to influence the brain sulfatide concentration and the activity and synthesis of an important enzyme involved in brain sphingolipids biosynthesis. Taken together, previous research results point to a possible role of vitamin K in the nervous system, especially during its development. Hence, the knowledge of the biologic role of vitamin K in the brain may be important for unveiling the mechanisms of normal and pathologic development and aging of the nervous system. The role of the vitamin K-dependent protein Gas6 in activation of signal transduction events in the brain in light of the age-related changes in the nervous system is also discussed.


Structural Analysis and Inhibition of TraE from the pKM101 Type IV Secretion System.
Gram-negative bacteria use type IV secretion systems (T4SSs) for a variety of macromolecular transport processes that include the exchange of genetic material. The pKM101 plasmid encodes a T4SS similar to the well-studied model systems from Agrobacterium tumefaciens and Brucella suis Here, we studied the structure and function of TraE, a homolog of VirB8 that is an essential component of all T4SSs. Analysis by X-ray crystallography revealed a structure that is similar to other VirB8 homologs but displayed an altered dimerization interface. The dimerization interface observed in the X-ray structure was corroborated using the bacterial two-hybrid assay, biochemical characterization of the purified protein, and in vivo complementation, demonstrating that there are different modes of dimerization among VirB8 homologs. Analysis of interactions using the bacterial two-hybrid and cross-linking assays showed that TraE and its homologs from Agrobacterium, Brucella, and Helicobacter pylori form heterodimers. They also interact with heterologous VirB10 proteins, indicating a significant degree of plasticity in the protein-protein interactions of VirB8-like proteins. To further assess common features of VirB8-like proteins, we tested a series of small molecules derived from inhibitors of Brucella VirB8 dimerization. These molecules bound to TraE in vitro, docking predicted that they bind to a structurally conserved surface groove of the protein, and some of them inhibited pKM101 plasmid transfer. VirB8-like proteins thus share functionally important sites, and these can be exploited for the design of specific inhibitors of T4SS function.


Induction and regulation of casein kinase II during B lymphocyte activation.
This study evaluates the regulation of casein kinase II (CK II) activity in resting B cells induced to enter the cell cycle. The induction of B cell cycle progression PMA and ionomycin results in an oscillatory expression of CK II. This kinase activity is also elicited after direct physical interaction between B cells and activated, fixed Th cells, indicating that the increase seen in CK II activity is probably associated with the delivery of the competence-inducing signal to resting B cells. The selective inhibition of ornithine decarboxylase (ODC), the rate-limiting enzyme for polyamine biosynthesis, during PMA and ionomycin-induction of B cell cycle progression, inhibits the expression of CK II activity. The addition of polyamines to cytosolic preparations recovered from cells in which ODC is inhibited results in the appearance of CK II activity, showing that the ODC inhibitor does not directly inhibit the kinase. The treatment of B cells with cycloheximide results in the appearance of CK II activity within 15 min, and this induction is partially explainable by a cycloheximide-elicited increase in cellular levels of polyamines. The artificial elevation of cellular levels of cAMP simultaneous with the addition of PMA and ionomycin results in a 150 to 200% increase in detectable CK II levels, suggesting that the cAMP-dependent signaling cascade may participate during the early regulation of CK II. In contrast, the inhibition of protein kinase C does not adversely influence the early expression of CK II, while actually enhancing kinase activity by 18 h poststimulation.


[Choroidal metastases as the form of presentation of 2 bronchiolo-alveolar carcinomas].
We report here two cases presenting with visual symptoms secondary to choroidal metastases, without any other remarkable systemic symptom. After an exhaustive investigation in both patients the diagnosis was obtained of bronchoalveolar carcinoma as primary tumor responsible for such lesions. Bronchoalveolar carcinoma is a type of pulmonary adenocarcinoma which rarely begins with clinical symptoms attributable to metastases and even rarer of choroidal location. The presence of such choroidal lesions are usually associated with visual clinical symptoms and they are a diagnostic challenge when manifested as the presenting symptoms. Therapy is directed to the primary tumor usually associated with palliative radiotherapy of choroidal metastases. Their presence implies a poor survival rate.


Creatively Adapting Touch-Based Practices to the Web Format During the COVID-19 Pandemic: Systematic Review.
The COVID-19 pandemic forced numerous touch-based fields, including manual therapy, body psychotherapy, arts therapy, creative arts practices, and the fitness industry, to swiftly transition to web-based service delivery. These disciplines faced substantial challenges in adapting their traditionally in-person practices, which rely heavily on physical touch and close proximity, to a web format. This review intends to provide a systematically sourced overview of the literature concerning innovative approaches for adapting touch-based practices to the web format in response to the COVID-19 pandemic. A systematic search across 7 databases and gray literature sources identified studies presenting innovative web delivery methods, particularly those addressing the challenges arising from the absence of physical proximity and touch. The inclusion criteria were designed to encompass studies exploring the creative adaptation of touch-based practices to web formats in response to the COVID-19 pandemic irrespective of study methodology. The exclusion criteria applied to studies focusing solely on technical aspects of web delivery or nontouch or noninteractive practices. There were no geographical restrictions, but the selection was limited to publications from 2020 onward. As only qualitative studies were found, data synthesis was conducted thematically. The review encompassed 17 studies revealing that touch-based fields successfully devised innovative and creative methods for web service delivery. These methods were categorized into five main themes: (1) adapted working methods (cross-field methods), (2) adapted working methods for sensorial experiences and nonphysical connections, (3) creative methods using materials or additional tools, (4) creative use of web-based platform tools or additional technologies, and (5) creative methods requiring previous preparation of practitioners or participants. These creative approaches allowed practitioners to address the challenges of web touch-based practices, fostering connections and offering unique sensory experiences, albeit with some concerns related to technology and preparation. These innovative methods demonstrate promise in circumventing the limitations imposed by the lack of physical touch and proximity in web settings during the COVID-19 pandemic. Furthermore, these insights hold potential for application in other fields in the future. This systematic search and thematic synthesis provide valuable guidance for practitioners and educators seeking to navigate the evolving landscape of web service delivery in touch-based disciplines, ensuring continuity of care even in challenging circumstances. PROSPERO CRD42022379731; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=379731.


Rational basis for the choice of regimens for empirical therapy of sepsis in granulocytopenic patients. A report of the EORTC International Antimicrobial Therapy Project Group.
The three therapeutic trials of the EORTC International Antimicrobial Therapy Project Group are reviewed, demonstrating the efficacy of synergistic combinations of aminoglycosides with beta-lactam compounds in the initial empirical therapy of febrile episodes in granulocytopenic patients. Large numbers of patients are required in order to have sufficient evaluable cases in small but important sub-groups to draw any valid and statistically significant conclusions. At present this can be achieved only by co-operative groups engaging in multi-centre trials.


Exploring Abstract Pattern Representation in The Brain and Non-symbolic Neural Networks.
Human cognitive and linguistic generativity depends on the ability to identify abstract relationships between perceptually dissimilar items. Marcus et al. (1999) found that human infants can rapidly discover and generalize patterns of syllable repetition (reduplication) that depend on the abstract property of identity, but simple recurrent neural networks (SRNs) could not. They interpreted these results as evidence that purely associative neural network models provide an inadequate framework for characterizing the fundamental generativity of human cognition. Here, we present a series of deep long short-term memory (LSTM) models that identify abstract syllable repetition patterns and words based on training with cochleagrams that represent auditory stimuli. We demonstrate that models trained to identify individual syllable trigram words and models trained to identify reduplication patterns discover representations that support classification of abstract repetition patterns. Simulations examined the effects of training categories (words vs. patterns) and pretraining to identify syllables, on the development of hidden node representations that support repetition pattern discrimination. Representational similarity analyses (RSA) comparing patterns of regional brain activity based on MRI-constrained MEG/EEG data to patterns of hidden node activation elicited by the same stimuli showed a significant correlation between brain activity localized in primarily posterior temporal regions and representations discovered by the models. These results suggest that associative mechanisms operating over discoverable representations that capture abstract stimulus properties account for a critical example of human cognitive generativity.


Association Between Concussion and Lower Extremity Injuries in Collegiate Athletes.
Concussions have been associated with elevated musculoskeletal injury risk; however, the influence of unreported and unrecognized concussions has not been investigated. The purpose of this study was to examine the association between concussion and lower extremity musculoskeletal injury rates across a diverse array of sports among collegiate student-athletes at the conclusion of their athletic career. The hypothesis was that there will be a positive association between athletes who reported a history of concussions and higher rates of lower extremity injuries. Cross-sectional study. Level 3. Student-athletes (N = 335; 62.1% women; mean age, 21.2 ± 1.4 years) from 13 sports completed a reliable injury history questionnaire. Respondents indicated the total number of reported, unreported, and potentially unrecognized concussions as well as lower extremity injuries including ankle sprains, knee injuries, and muscle strains. Chi-square analyses were performed to identify the association between concussion and lower extremity injuries. There were significant associations between concussion and lateral ankle sprain ( P = 0.012), knee injury ( P = 0.002), and lower extremity muscle strain ( P = 0.031). There were also significant associations between reported concussions and knee injury ( P = 0.003), unreported concussions and knee injury ( P = 0.002), and unrecognized concussions and lateral ankle sprain ( P = 0.001) and lower extremity muscle strains ( P = 0.006), with odds ratios ranging from 1.6 to 2.9. There was a positive association between concussion history and lower extremity injuries (odds ratios, 1.6-2.9 elevated risk) among student-athletes at the conclusion of their intercollegiate athletic careers. Clinicians should be aware of these elevated risks when making return-to-participation decisions and should incorporate injury prevention protocols.


Morphological studies on machined implants of commercially pure titanium and titanium alloy (Ti6Al4V) in the rabbit.
The aim of this study was to evaluate the bone response to commercially pure titanium grade I and titanium alloy grade V (90% Ti, 6% Al, and 4% V, depicted Ti6Al4V) after 8 weeks in rabbit tibia. Interference microscopy and scanning electron microscopy were used for surface analyses. Transmission electron microscopy (TEM) was used for evaluation of surface crystallinity and chemistry after preparation of ultrathin sections using focused ion beam (FIB) microscopy. Three different embedding resins commonly used for histological preparation were evaluated with respect to adaptation to a turned implant surface. Epoxy Agar 100 resin and acrylic Technovit 7200 resin showed low separation while acrylic LR White resin showed large separation at the interface. The retrieved specimens were embedded in acrylic Technovit 7200 resin after fixation and dehydration. The histological evaluation revealed osseointegration for both c.p. titanium grade I and Ti6Al4V alloy, but no quantitative differences in bone contact and bone area were detected. Because a separation of implant and tissue occurred in the interface between implant and bone embedded in acrylic Technovit 7200 resin, additional factors related to implant surface properties and technical procedures are likely to influence the possibilities to prepare ultrathin sections by FIB.


Numerical and DNA: DNA reassociation analyses of Erwinia rubrifaciens and other members of the Enterobacteriaceae.
Phenetic data on 75 strains of Erwinia and other representative genera of the Enterobacteriaceae were collected and analysed using two numerical taxonomic methods. In both methods the same subclusters were recovered. The subclusters, however, were defined at different similarity levels and were classified into clusters of different composition. Erwinia rubrifaciens strains formed a very tight, homogeneous subcluster, completely distinct and readily distinguishable from other Erwinia and enterobacteria species studied. DNA: DNA hybridization between E. rubrifaciens and Erwinia and Shigella species were analysed to corroborate the numerical classifications. A good correlation between the numerical and DNA: DNA hybridization analyses was found and provided sufficient evidence for not supporting the previously proposed subspecific taxonomic position of E. rubrifaciens. The data clearly showed that E. rubrifaciens is a separate species in its own right. Based on the existence of very high genetic relatedness and high similarities in phenetic characters among E. rubrifaciens strains and the confinement of the pathogen to the state of California, the hypothesis is offered that E. rubrifaciens originated from a single source.


Individual recovery profiles in basketball players.
In the sport context, recovery has been characterized as a multifactor process (physiological, psychological, behavioral, social, etc.). This study takes a multidisciplinary approach to find psychophysiological markers of the stress-recovery process. It aims to determine how athletes' specific recovery actions relate to their perceptions of recovery, and Heart Rate Variability (HRV). A total of 196 assessments were analyzed from 6 players on a men's professional basketball team within the Liga LEB Oro basketball federation (2012/2013 season). Perceptions of recovery, recovery strategies, and HRV were recorded. The results show a pattern of individual differences in behavior related to athletes' recovery actions and HRV profiles throughout the season (p < .05). Moreover, we observed that each player had different recovery needs. In light of these results, we suggest an individualistic approach to evaluating and monitoring recovery to attend more accurately to each player's recovery needs.


Klotho, phosphate and FGF-23 in ageing and disturbed mineral metabolism.
High concentrations of extracellular phosphate are toxic to cells. Impaired urinary phosphate excretion increases serum phosphate level and induces a premature-ageing phenotype. Urinary phosphate levels are increased by dietary phosphate overload and might induce tubular injury and interstitial fibrosis. Extracellular phosphate exerts its cytotoxic effects by forming insoluble nanoparticles with calcium and fetuin-A; these nanoparticles are referred to in this Review as calciprotein particles. Calciprotein particles are highly bioactive ligands that can induce various cellular responses, including the osteogenic transformation of vascular smooth muscle cells and cell death of vascular endothelial cells and renal tubular epithelial cells. Calciprotein particles are detected in the serum of animal models of kidney disease and in patients with chronic kidney disease (CKD) and might be associated with a (mal)adaptation of the endocrine axes mediated by fibroblast growth factors and Klothos that regulate phosphate homeostasis and ageing. These observations raise the possibility that calciprotein particles contribute to the pathogenesis of CKD. This theory, if verified, is expected to provide novel diagnostic markers and therapeutic targets in CKD.


Enhancing Bioproducts in Seaweeds via Sustainable Aquaculture: Antioxidant and Sun-Protection Compounds.
Marine macroalgae are considered an untapped source of healthy natural metabolites and their market demand is rapidly increasing. Intertidal macroalgae present chemical defense mechanisms that enable them to thrive under changing environmental conditions. These intracellular chemicals include compounds that can be used for human benefit. The aim of this study was to test cultivation protocols that direct seaweed metabolic responses to enhance the production of target antioxidant and photoprotective biomaterials. We present an original integrated multi-trophic aquaculture (IMTA) design, based on a two-phase cultivation plan, in which three seaweed species were initially fed by fish effluents, and subsequently exposed to various abiotic stresses, namely, high irradiance, nutrient starvation, and high salinity. The combined effect of the IMTA's high nutrient concentrations and/or followed by the abiotic stressors enhanced the seaweeds' content of mycosporine-like amino acids (MAAs) by 2.3-fold, phenolic compounds by 1.4-fold, and their antioxidant capacity by 1.8-fold. The Sun Protection Factor (SPF) rose by 2.7-fold, and the chlorophyll and phycobiliprotein synthesis was stimulated dramatically by an order of magnitude. Our integrated cultivation system design offers a sustainable approach, with the potential to be adopted by emerging industries for food and health applications.


Mitosis-specific phosphorylation of vimentin by protein kinase C coupled with reorganization of intracellular membranes.
Using two types of anti-phosphopeptide antibodies which specifically recognize vimentin phosphorylated by protein kinase C (PKC) at two distinct PKC sites, we found that PKC acted as a mitotic vimentin kinase. Temporal change of vimentin phosphorylation by PKC differed form changes by cdc2 kinase. The mitosis-specific vimentin phosphorylation by PKC was dramatically enhanced by treatment with a PKC activator, 12-O-tetradecanoylphorbol-13-acetate (TPA), while no phosphorylation of vimentin by PKC was observed in interphase cells treated with TPA. By contrast, the disruption of subcellular compartmentalization of interphase cells led to vimentin phosphorylation by PKC. Cytoplasmic and nuclear membranes are fragmented and dispersed in the cytoplasm and some bind to vimentin during mitosis. Thus, targeting of activated PKC, coupled with the reorganization of intracellular membranes which contain phospholipids essential for activation, leads to the mitosis-specific phosphorylation of vimentin. We propose that during mitosis, PKC may phosphorylate an additional subset of proteins not phosphorylated in interphase.


Real-time measurements of local myocardium motion and arterial wall thickening.
We have already developed a new method, namely, the phased tracking method, to track the movement of the heart wall and arterial wall accurately based on both the phase and magnitude of the demodulated signals to determine the instantaneous position of an object. This method has been realized by an off-line measurement system, which cannot be applied to transient evaluation of rapid response of the cardiovascular system to physiological stress. In this paper, therefore, a real-time system to measure change in the thickness of the myocardium and the arterial wall is presented. In this system, an analytic signal from standard ultrasonic diagnostic equipment is analogue-to-digital (A/D) converted at a sampling frequency of 1 MHz. By pipelining and parallel processing using four high-speed digital signal processing (DSP) chips, the method described is realized in real time. The tracking results for both sides of the heart and/or arterial wall are superimposed on the M (motion)-mode image in the work station (WS), and the thickness changes of the heart and/or arterial wall are also displayed and digital-to-analogue (D/A) converted in real time. From the regional change in thickness of the heart wall, spatial distribution of myocardial motility and contractility can be evaluated. For the arterial wall, its local elasticity can be evaluated by referring to the blood pressure. In in vivo experiments, the rapid response of the change in wall thickness of the carotid artery to the dose of the nitroglycerine (NTG) is evaluated. This new real-time system offers potential for quantitative diagnosis of myocardial motility, early stage atherosclerosis, and the transient evaluation of the rapid response of the cardiovascular system to physiological stress.


Improving Molecular and Histopathology in Diaphragm Muscle of the Double Transgenic ACTA1-MCM/FLExDUX4 Mouse Model of FSHD with Systemic Antisense Therapy.
Facioscapulohumeral muscular dystrophy (FSHD) is a rare muscle dystrophy causing muscle weakness initially in the face, shoulders, and upper arms, and extends to lower body muscles as the disease progresses. Respiratory restriction in FSHD is increasingly reported to be more common and severe than previously thought, with the involvement of diaphragm weakness in pulmonary insufficiency being under debate. As aberrant expression of the double homeobox 4 (DUX4) gene is the prime cause of FSHD, we and others have developed numerous strategies and reported promising results on downregulating DUX4 expression in both cellular and animal models of FSHD. However, the effect of DUX4 and anti-DUX4 approaches on diaphragm muscle has not been elucidated. In this study, we show that toxic DUX4 expression causes pathology that affects the diaphragm of ACTA1-MCM/FLExDUX4 mouse model of FSHD at both molecular and histological levels. Of importance, a systemic antisense treatment that suppresses DUX4 and target genes expression by 50% significantly improves muscle regeneration and muscle fibrosis, and prevents modification in myofiber type composition, supporting its development as a treatment for FSHD.


Patterns of growth and development in narcotic-exposed children.
The results of the studies reviewed indicate that intrauterine growth is adversely affected by drug use during pregnancy. Whether the impairment is a direct effect of narcotic exposure or is the result of the interaction of deleterious health, environmental, and socioeconomic factors closely associated with the lifestyle of the woman who abuses drugs cannot be determined at present. Reports on the long-term effects of drug use on growth and intellectual functioning in the offspring of women who abuse drugs are not consistent. While some studies indicate that most of the exposed infants exhibit catchup growth by 6 months of age (Lifschitz et al. 1983, 1985), one methodologically strong study suggests that methadone may have a small direct teratogenic effect reflected in reduced head size at 2 years of age (Hans 1989). Unexplained is the pattern of growth deceleration observed in some narcotic-exposed children (Lifschitz et al. 1983, 1985). The few available reports on long-term outcome concur that narcotic-exposed children have a high incidence of behavioral and learning problems (Strauss et al. 1979; Rosen and Johnson 1985; Wilson 1989), but population studies have been too small to demonstrate that they differ significantly from controls. There is a suggestion that narcotic use during pregnancy promotes a biological vulnerability to adverse environments, manifested in the neurobehavioral and intellectual areas.


Clinical utility of molecular and flow cytometric markers in chronic lymphocytic leukaemia.
Chronic lymphocytic leukaemia (CLL) is a clinically heterogeneous disease. While immunoglobulin variable region heavy chain (IgVH) mutational status remains the 'gold standard' in molecular prognostication, a range of additional markers is increasingly being used in clinical trials. As awareness of trial data increases, requests to determine these prognostic markers for new CLL patients are becoming more prevalent in Australia. To explore the clinical utility of currently available prognostic markers for CLL in an Australian cohort. IgVH mutational status and gene usage was determined and compared with other reported immunophenotypic markers, cytogenetics and clinical outcome as defined by treatment-free survival (TFS), lymphocyte doubling time and clinical stage in a cohort of 65 CLL patients. An unmutated IgVH gene, high expression of CD38, ZAP-70, CD25, CD49d, CD54 or low expression of CD49c was associated with shorter TFS indicating an adverse clinical prognosis in our cohort. High expression of each of CD38, ZAP-70, CD49d and CD54 was significantly associated with an unmutated IgVH gene; however, associations were not absolute. IgVH and CD25 expression retained their significance in multivariate analysis. Concordant CD25(high) /IgVH unmutated CLL patients had the shortest median TFS interval (40 months) in our cohort. Molecular and immunophenotypic markers remain useful as adjuncts to clinical prognostication; however, as single parameters they are unable to dictate the timing of therapeutic intervention. The combined use of CD25 and IgVH mutational status may be clinically relevant to CLL prognostication while also providing insight into the biological pathways involved in disease progression.


Vascular endothelial growth factor mediates angiogenic activity during the proliferative phase of wound healing.
Angiogenesis is an essential component of normal wound repair, yet the primary mediators of wound angiogenesis have not been well described. The current study characterizes the contribution of vascular endothelial cell growth factor (VEGF) to the angiogenic environment of human surgical wounds. Surgical wound fluid samples (n = 70) were collected daily for up to 7 postoperative days (POD) from 14 patients undergoing mastectomy or neck dissection. VEGF levels in surgical wound fluid were lowest on POD 0, approximating values of serum, but increased steadily through POD 7. An opposite pattern was noted for basic fibroblast growth factor-2. Fibroblast growth factor-2, which has been previously described as a wound angiogenic factor, exhibited highest levels at POD 0, declining to near serum levels by POD 3. Surgical wound fluid form all time points stimulated marked endothelial cell chemotaxis and induced a brisk neovascular response in the rat corneal micropocket angiogenesis assay. Antibody neutralization of VEGF did not affect the in vitro chemotactic or the in vivo angiogenic activity early wound samples (POD 0). In contrast, VEGF neutralization significantly attenuated both chemotactic activity (mean decrease 76 +/- 13%, P < 0.01) and angiogenic activity (5 of 5 samples affected) of later wound samples (POD 3 and 6). The results suggest a model of wound angiogenesis in which an initial angiogenic stimulus is supplied by fibroblast growth factor-2, followed by a subsequent and more prolonged angiogenic stimulus mediated by VEGF.


Copper II - polar amino acid complexes: toxicity to bacteria and larvae of Aedes aegypti.
Control strategies using insecticides are sometimes ineffective due to the resistance of the insect vectors.In this scenario new products must be proposed for the control of insect vectors.The complexes L-aspartate Cu (II) and L-glutamate-Cu (II) complexes were synthesized and characterized by elemental analysis, visible ultraviolet, infrared spectroscopy and potentiometric titration. The toxicity of these complexes was analyzed in Aedes aegypti (Diptera: Culicidae) larvae and Gram-negative and Gram-positive bacteria. The interaction between the ligands and the amino acid balance and the distribution of the species as a function of pH were discussed. The lethal concentration median (LC50) for Ae. aegypti larvae were: L-glutamic acid-Cu (II) - 53.401 mg L-1 and L-aspartate-Cu (II) - 108.647 mg L-1. The minimum inhibitory concentration (MIC) required for Staphylococcus aureus and Escherichia coli was: L-glutamate-Cu (II) 500-2000 mg L-1 and L-aspartate-Cu (II) 1000-2000 mg L-1. The concentrations demonstrated toxicity that evidence the potential of the complexes as bactericide and insecticide. Metal complexes formed by amino acids and transition metals are advantageous because of low environmental toxicity, biodegradability and low production cost.


Generic substitutions: a 2005 survey of the acceptance and perceptions of physicians in Jamaica.
In Jamaica, the 1993 amendment to the Pharmacy Act allows pharmacists to offer patients generic substitutions for innovator brands; however, there were reservations among physicians about this policy implementation. The success of the amendment may be influenced by the confidence of physicians in the therapeutic equivalence of generics, especially since the act also allows physicians to indicate "no substitution" on prescriptions. The aim of this investigation was to examine the current attitudes of physicians towards the use of generic substitutions. One hundred questionnaires were distributed island-wide among physicians of varying specialities, with items to characterize their demographics and specific statements to determine their perception of generics. Sixty questionnaires were returned completed (60% response). Most of the responding physicians were males (2:1 male:female ratio); the majority were general/family medicine physicians in private practice. Forty-nine per cent of the responding physicians were mostly prescribing generic brands willingly, indicating that the cheaper cost of generic substitutes was a significant factor for this trend. There were doubts about whether bioequivalence of a generic was equitable to therapeutic equivalence to innovator drug. Additionally, 33% of the physicians were able to identify at least one case in the past year of clinical problems with generic substitutes that they perceived would not have occurred with the innovator. It is concluded that while the amendment to the Pharmacy Act encourages the substitution of generics in preference to innovator brands, more emphasis should be placed on improving physician confidence in the therapeutic equivalence of generics.


TGFbeta induces SIK to negatively regulate type I receptor kinase signaling.
Signal transduction by transforming growth factor beta (TGFbeta) coordinates physiological responses in diverse cell types. TGFbeta signals via type I and type II receptor serine/threonine kinases and intracellular Smad proteins that regulate transcription. Strength and duration of TGFbeta signaling is largely dependent on a negative-feedback program initiated during signal progression. We have identified an inducible gene target of TGFbeta/Smad signaling, the salt-inducible kinase (SIK), which negatively regulates signaling together with Smad7. SIK and Smad7 form a complex and cooperate to down-regulate the activated type I receptor ALK5. We further show that both the kinase and ubiquitin-associated domain of SIK are required for proper ALK5 degradation, with ubiquitin functioning to enhance SIK-mediated receptor degradation. Loss of endogenous SIK results in enhanced gene responses of the fibrotic and cytostatic programs of TGFbeta. We thus identify in SIK a negative regulator that controls TGFbeta receptor turnover and physiological signaling.


Induced expression of CCL19 promotes the anti-tumor ability of CAR-T cells by increasing their infiltration ability.
Chimeric antigen receptor-engineered T cell (CAR-T) therapy has shown promising potential for anti-cancer treatment. However, for pancreatic ductal adenocarcinoma (PDAC), the lack of infiltrative ability of these CAR-T cells leads to sub-optimal treatment outcome. Chemokine (C-C motif) ligand 19 (CCL19), the expression of which is regulated by the nuclear factor of activated T cell pathway, was transfected into targeting mesothelin CAR-T cells (mesoCAR-N19) using NFAT regulating element. It was expressed in activated CAR-T cells by OKT3 or mesothelin+ tumor cells but not in inactive cells. The migratory ability of these CAR-T cells was then measured. Subsequently, functional identification of these CAR-T cells was performed in vivo. In addition, the tumor lytic activity and proliferation of the CAR-T cells were measured in vitro. The degree of CAR-T cell infiltration and distribution into the PDAC tumors was examined using the immunohistochemical staining of hCD3 and the detection of CAR gene copy number by quantitative PCR. Finally, the functional assessment of chemokine (C-C motif) receptor 7 knock-out was performed in the CAR-T cells. Through in vitro Transwell assays, it was demonstrated that mesoCAR-N19 can be specifically expressed in CAR-T cells activated by tumor cells compared with conventional mesothelin CAR-T (mesoCAR) cells. We also observed that upregulating the expression of CCL19 can increase the recruitment of additional T cells. In vivo studies subsequently revealed that this highly specific recruitment of T cell infiltration is associated with enhanced tumor-suppressive activities downstream. Induced expression of CCL19 can promote the anti-tumor ability of CAR-T cells by increasing their infiltrative ability. This study potentially uncovered novel method of activating CAR-T cells to enhance their infiltrative capacities, which offers a novel direction for PDAC treatment.


Decreasing pelvic incidence is associated with greater risk of cam morphology.
The spinopelvic relationship (including pelvic incidence) has been shown to influence pelvic orientation, but its potential association with femoroacetabular impingement has not been thoroughly explored. The purpose of this study was to prove the hypothesis that decreasing pelvic incidence is associated with increased risk of cam morphology. Two matching cohorts were created from a collection of cadaveric specimens with known pelvic incidences: 50 subjects with the highest pelvic incidence (all subjects > 60°) and 50 subjects with the lowest pelvic incidence (all subjects < 35°). Femoral version, acetabular version, and alpha angles were directly measured from each specimen bilaterally. Cam morphology was defined as alpha angle > 55°. Differences between the two cohorts were analysed with a Student's t-test and the difference in incidence of cam morphology was assessed using a chi-squared test. The significance level for all tests was set at p < 0.05. Cam morphology was identified in 47/100 (47%) femurs in the cohort with pelvic incidence < 35° and in only 25/100 (25%) femurs in the cohort with pelvic incidence > 60° (p = 0.002). The mean alpha angle was also greater in the cohort with pelvic incidence < 35° (mean 53.7°, sd 10.7° versus mean 49.7°, sd 10.6°; p = 0.008). Decreased pelvic incidence is associated with development of cam morphology. We propose a novel theory wherein subjects with decreased pelvic incidence compensate during gait (to maintain optimal sagittal balance) through anterior pelvic tilt, creating artificial anterior acetabular overcoverage and recurrent impingement that increases risk for cam morphology.Cite this article: W. Z. Morris, C. A. Fowers, R. T. Yuh, J. J. Gebhart, M. J. Salata, R. W. Liu. Decreasing pelvic incidence is associated with greater risk of cam morphology. Bone Joint Res 2016;5:387-392. DOI: 10.1302/2046-3758.59.BJR-2016-0028.R1.


Metal-dependent inhibition of amyloid fibril formation: synergistic effects of cobalt-tannic acid networks.
Metal-phenolic networks (MPNs) have received widespread interest owing to their modular incorporation of functional metal ions and phenolic ligands. However, the interaction between MPNs and biomolecules is still relatively unexplored. Herein, we studied the effects of MPN-coated gold nanoparticles on amyloid fibril formation (which is associated with Alzheimer's disease) as a function of the metal ion in the MPN systems. All coated particles examined inhibited amyloid formation, with cobalt(ii) MPN-coated particles exhibiting the highest inhibition activity (90%). Molecular dynamics simulations and quantum mechanics calculations suggested that the geometry of the exposed cobalt coordination site in the cobalt-tannic acid networks facilitates its interactions with histidine and methionine residues in the amyloid beta peptides. Furthermore, the unique structure of cobalt MPNs may enable a wider variety of biomedical applications.


Performance of six functionals (LDA, PBE, PBESOL, B3LYP, PBE0, and WC1LYP) in the simulation of vibrational and dielectric properties of crystalline compounds. The case of forsterite Mg2SiO4.
The performance of six different density functionals (LDA, PBE, PBESOL, B3LYP, PBE0, and WC1LYP) in describing the infrared spectrum of forsterite, a crystalline periodic system with orthorhombic unit cell (28 atoms in the primitive cell, Pbmn space group), is investigated by using the periodic ab initio CRYSTAL09 code and an all-electron Gaussian-type basis set. The transverse optical (TO) branches of the 35 IR active modes are evaluated at the equilibrium geometry together with the oscillator strengths and the high-frequency dielectric tensor ϵ(∞) . These quantities are essential to compute the dielectric function ϵ(ν), and then the reflectance spectrum R(ν), which is compared with experiment. It turns out that hybrid functionals perform better than LDA and GGA, in general; that B3LYP overperforms WC1LYP and, in turn, PBE0; that PBESOL is better than PBE; that LDA is the worst performing functional among the six under study.


Telephone contact intervention in women undergoing treatment for breast cancer.
The chemotherapy received by breast cancer patients is complicated by many side effects. At our centre, a health care professional is accessible at all times, yet we hypothesize that most breast cancer patients suffer in silence rather than phone for assistance. This study sought to assess the value of and perceived need for a telephone call to breast cancer patients following the initiation of chemotherapy. The women's side effects were also documented, and the level to which they were educated about symptoms to expect and available assistance was evaluated. Overall, the weekend calls were well received because they provided information and support to the patients. It was determined that further research is required to determine if and how such a call should be best implemented at our cancer centre.


ATM protein is required for radiation-induced apoptosis and acts before mitochondrial collapse.
To define the role of the ataxia telangiectasia (A-T) mutated gene (ATM) in activation and progress of apoptosis. Three normal and three A-T EBV-transformed cell lines were studied. Following irradiation (IR), Fas activation or ceramide exposure, viability and apoptosis were measured by trypan blue dye exclusion assay and as sub-G1 cell fraction by flow cytometric analysis of propidium iodide stained cultures, respectively. Activation of caspase-3 was evaluated by immunoblot and by an in vitro activity assay on cytosolic cell extracts. To assess changes in mitochondrial potential and reactive oxygen species, cells were stained by 3,3'-dihexyloxacarbocynine iodide or hydroethidine, respectively, and scored by flow cytometry. The observations establish that A-T cells are equipped with a proficient apoptotic machinery, as demonstrated by their ability to undergo mitochondrial collapse and caspase-3 activation after Fas activation or ceramide treatment. Both treatments have a similar cytotoxic effect on normal and A-T cells. In contrast, in spite of the stronger cytotoxicity induced by IR exposure, irradiated A-T cells are unable to undergo mitochondrial collapse and caspase-3 activation. The data indicate that ATM is necessary in the initiation of molecular pathway(s) leading to IR-induced apoptosis, and suggest that increased radiosensitivity of A-T cells is more likely a direct consequence of necrotic cell death.


Psychiatric Mental Health Leadership at the Tipping Point.
Currently the United States health care system is responding to the Patient Protection and Affordable Care Act (PPACA) and the vision it contains for health care transformation. Along with sweeping changes in service delivery and payment structures, health care reform has championed concepts such as patient-centered care, integrated care, and wellness. Although these are not new ideas, their adaptation, in both ideology and service design has been accelerated in the context for reform. Indeed they are reaching a tipping point; the point where ideas gain wide acceptance and become influential trends. Although psychiatric mental health (PMH) nurses have been active in wellness, patient-centered care, and integrated care, at the current time they seem to be situated peripheral to these national trends. Increased presence of PMH nurses will facilitate their contribution to the development of these concepts within service structures and interventions. To increase knowledge and appreciation of PMH nurses' practice and unique perspective on these issues, leaders are needed who will connect and effectively communicate PMH nursing efforts to the broader health care arena. This article outlines the events that created a context for these three concepts (patient-centered care, wellness, and integrated care), and I suggest why they have reached a tipping point and discuss the need for greater PMH nursing presence in the American national dialog and the role of nursing leaders in facilitating these connections.


Fatal iatrogenic cerebral β-amyloid-related arteritis in a woman treated with lecanemab for Alzheimer's disease.
We report the case of a 79-year-old woman with Alzheimer's disease who participated in a Phase III randomized controlled trial called CLARITY-AD testing the experimental drug lecanemab. She was randomized to the placebo group and subsequently enrolled in an open-label extension which guaranteed she received the active drug. After the third biweekly infusion, she suffered a seizure characterized by speech arrest and a generalized convulsion. Magnetic resonance imaging revealed she had multifocal swelling and a marked increase in the number of cerebral microhemorrhages. She was treated with an antiepileptic regimen and high-dose intravenous corticosteroids but continued to worsen and died after 5 days. Post-mortem MRI confirmed extensive microhemorrhages in the temporal, parietal and occipital lobes. The autopsy confirmed the presence of two copies of APOE4, a gene associated with a higher risk of Alzheimer's disease, and neuropathological features of moderate severity Alzheimer's disease and severe cerebral amyloid angiopathy with perivascular lymphocytic infiltrates, reactive macrophages and fibrinoid degeneration of vessel walls. There were deposits of β-amyloid in meningeal vessels and penetrating arterioles with numerous microaneurysms. We conclude that the patient likely died as a result of severe cerebral amyloid-related inflammation.


Pediatric Cancer in Northern Tanzania: Evaluation of Diagnosis, Treatment, and Outcomes.
The majority of new diagnoses of pediatric cancer are made in resource-poor countries, where survival rates range from 5% to 25% compared with 80% in high-resource countries. Multiple factors, including diagnostic and treatment capacities and complex socioeconomic factors, contribute to this variation. This study evaluated the available resources and outcomes for pediatric patients with cancer at the first oncology treatment center in northern Tanzania. Qualitative interviews were completed from July to August 2015 to determine available staff, hospital, diagnostic, treatment, and supportive care resources. A retrospective review of hospital admissions and clinic visits from January 2010 to August 2014 was completed. A total of 298 patients were identified, and data from 182 patient files were included in this review. Diagnostic, treatment, and supportive capacities are limited for pediatric cancer care. The most common diagnoses were Burkitt lymphoma (n = 32), other non-Hodgkin lymphoma (n = 26), and Wilms tumor (n = 25). A total of 40% of patients (n = 72) abandoned care. There was a 20% 2-year event-free survival rate, which was significantly affected by patient age, method of diagnosis, and year of diagnosis. To our knowledge, this is the first review of pediatric cancer outcomes in northern Tanzania. The study identified areas for future development to improve pediatric cancer outcomes, which included strengthening of training and diagnostic capacities, development of registries and research databases, and the need for additional research to reduce treatment abandonment.


Mannose-binding lectin and the risk of HIV transmission and disease progression in children: a systematic review.
Mannose-binding lectin (MBL) can activate the complement system by binding to carbohydrates, such as those presented on the HIV virion surface. It is unclear whether genetically determined MBL deficiency is related to vertical HIV transmission and disease progression in HIV-infected children. A literature search of Medline, Embase and Cochrane Central Register identified all relevant studies on MBL and HIV infection in children. We extracted information on the characteristics of the study group, method of MBL analysis, outcome definitions, follow-up and the risk estimates. The validity of each study was assessed. Nine studies were retrieved. Most were of good validity, but risk adjustment for confounders was missing in 6 studies. Age, treatment and outcome definitions differed between the study groups. In most of the studies, MBL deficiency was associated with an increased frequency of vertical HIV transmission and an increased speed of disease progression. In the 2 most valid studies, carriers of variant genes had an increased odds ratio for transmission and an increased relative hazard for disease progression and central nervous system impairment, especially in children <2 years of age. MBL deficiency is associated with an increased risk of vertical HIV transmission. How this risk relates to other factors that influence transmission is unclear. The association between HIV disease progression and MBL deficiency is most pronounced in children <2 years of age, probably due to immaturity of their adaptive immunity.


Sexual orientation, gender, and racial differences in illicit drug use in a sample of US high school students.
We evaluated drug use differences between sexual minority and heterosexual students, including interactions with gender and race/ethnicity. We used 2005 and 2007 Youth Risk Behavior Survey data pooled from Boston, Massachusetts; Chicago, Illinois; Delaware; Maine; Massachusetts; New York City, New York; Rhode Island; and Vermont to evaluate drug use (marijuana, cocaine, inhalants, heroin, methamphetamine, and MDMA [Ecstasy]) using 2 aspects of sexual orientation (identity and sex of sexual partners). Sexual minority students had higher prevalence of drug use than did heterosexuals on both sexual orientation dimensions, and differences were particularly pronounced among bisexual students on both dimensions. Differences between sexual minority and heterosexual male students in prevalence were generally larger than were differences between sexual minority and heterosexual female students. Racial minority students generally reported lower prevalence of drug use. However, the protective effect of African American race was less pronounced for some sexual minorities. Sexual minority youths are at increased risk for drug use. Intervention is needed at the institutional and individual levels to address these disparities.


A melt-electrowritten filter for capture and culture of circulating colon cancer cells.
Metastasis is the major cause of death in cancer patients accounting for about 90% of the mortality. The detection and analysis of the hallmark of metastasis, circulating tumor cells (CTCs), have significant impact in cancer biology and clinical practice. However, the scarcity of CTCs in blood, particularly in that of colorectal cancer patients, is a serious bottleneck in the development of CTC-based precision medicine. Herein, the melt electrowriting (MEW) technology was used for reproductive fabrication of a biocompatible antibody-presenting polycaprolactone filter with tailored porous structure. It is demonstrated, for the first time, that such filter can be used not only to catch cancer cells spiked in whole blood but also to culture the cancer cells directly on site. Specifically, HT29 colon cancer cells can be captured with an efficiency of 85%, and when spiked into 4 mL of whole blood, 47% were captured on one Ø12mm filter. Furthermore, repeated capture and culture experiments have shown that as few as 20 HT29 colon cancer cells spiked into 4 mL of whole blood can be captured on the filter and within 2 weeks be expanded on site to become tumor bodies that are visible to the untrained eye. This filter allows for downstream analysis, such as flow cytometry, immunocytochemistry, Western blotting, and rt-qPCR. This technology represents a simple and cost-effective platform that potentially enables fast and efficient culture of rare CTCs from patients' blood. This provides non-invasive alternatives for solid biopsy tumor materials for treatment screening, with great potential to realize precision medicine for cancer treatment.


The excitation cascade of Limulus ventral photoreceptors: guanylate cyclase as the link between InsP3-mediated Ca2+ release and the opening of cGMP-gated channels.
Early stages in the excitation cascade of Limulus photoreceptors are mediated by activation of Gq by rhodopsin, generation of inositol-1,4,5-trisphosphate by phospholipase-C and the release of Ca2+. At the end of the cascade, cGMP-gated channels open and generate the depolarizing receptor potential. A major unresolved issue is the intermediate process by which Ca2+ elevation leads to channel opening. To explore the role of guanylate cyclase (GC) as a potential intermediate, we used the GC inhibitor guanosine 5'-tetraphosphate (GtetP). Its specificity in vivo was supported by its ability to reduce the depolarization produced by the phosphodiesterase inhibitor IBMX. To determine if GC acts subsequent to InsP3 production in the cascade, we examined the effect of intracellular injection of GtetP on the excitation caused by InsP3 injection. This form of excitation and the response to light were both greatly reduced by GtetP, and they recovered in parallel. Similarly, GtetP reduced the excitation caused by intracellular injection of Ca2+. In contrast, this GC inhibitor did not affect the excitation produced by injection of a cGMP analog. We conclude that GC is downstream of InsP3-induced Ca2+ release and is the final enzymatic step of the excitation cascade. This is the first invertebrate rhabdomeric photoreceptor for which transduction can be traced from rhodopsin photoisomerization to ion channel opening.


Brain N-acetylaspartate as a molecular water pump and its role in the etiology of Canavan disease: a mechanistic explanation.
N-acetyl-L-aspartate (NAA), an abundant amino acid present in the vertebrate brain, is synthesized and stored primarily in neurons. Its metabolism is also dynamic, with NAA turning over more than once each day by its regulated efflux into extracellular fluid (ECF), cycling between an anabolic L-aspartate acetylating compartment in neurons and a catabolic NAA deacetylating compartment in oligodendrocytes. An inborn error in NAA metabolism results in Canavan disease (CD), a rare and usually fatal early-onset autosomal recessive human central nervous system (CNS) disease, caused by failure of the catabolic metabolism of NAA resulting from a lack of sufficient amidohydrolase II activity in oligodendrocytes. Various hypotheses regarding the metabolism of NAA and its role have been considered, and although NAA may perform several functions in the CNS, an important role of NAA appears to be osmoregulatory. Based on this role, an osmotic-hydrostatic mechanism for the etiology of the CD phenotype is proposed. In CD, the daily addition of 13375 Pascals (0.132 atmospheres or 1.94 lbs per square inch) of hydrostatic pressure to brain ECF, on the brain cell side of brain-barrier epithelial membranes, resulting from the continuous synthesis and efflux of NAA, is considered to be responsible for the syndrome.


Evidence of significant absorption of antipyrine from urinary bladder of rats.
We examined the in vivo absorption and pharmacokinetics of antipyrine, a base that is unionized at physiologic pH, from the urinary bladder of adult female Fischer rats. The clearance of an i.v. dose of antipyrine (25 mg/kg) was found to vary considerably between animals, which is consistent with the literature data. Therefore, it was necessary to simultaneously determine drug clearance and bladder absorption in the same animal. This was accomplished by giving concomitantly an i.v. dose of [14C]antipyrine (2.5 muCi, about 90 micrograms/kg) via a jugular vein catheter and an intravesical dose of unlabeled antipyrine (33 mg/kg) via a urethral catheter. Unlabeled antipyrine was detected in plasma, indicating the absorption of antipyrine into systemic circulation. The bioavailability of the intravesical dose was calculated using the clearance of [14C]antipyrine and the plasma concentrations of unlabeled antipyrine. The intravesical dose was withdrawn through the urethral catheter after 90 min. To minimize mechanical manipulation and damage, the bladder was not rinsed. This may have caused the incomplete recovery of the unabsorbed dose; about 65 +/- 18% (mean +/- S.D.) of the dose was recovered at 90 min. Maximal plasma concentrations were achieved at 10 to 48 min after removal of the intravesical dose, which is consistent with a continued absorption of the residual dose. The intravesical bioavailability was variable between animals, with an average of 11.6 +/- 6.4% (mean +/- S.D.; range, 4.1-19.2%). In conclusion, these data demonstrate that neutral drugs such as antipyrine are absorbed from the bladder, that the extent of absorption is variable and that the urinary bladder may be a site of significant re-entry of drugs into the systemic circulation.


Enhanced in vitro chondrogenesis of primary mesenchymal stem cells by combined gene transfer.
Because articular cartilage has a poor regeneration capacity, numerous cell-based approaches to therapy are currently being explored. The present study involved the use of gene transfer as a means to provide sustained delivery of chondrogenic proteins to primary mesenchymal stem cells (MSCs). In previous work, we found that adenoviral-mediated gene transfer of transforming growth factor-beta1 (TGF-beta1) and bone morphogenetic protein 2 (BMP-2), but not insulin-like growth factor 1 (IGF-1), could be used to induce chondrogenic differentiation of MSCs in an aggregate culture system. In the present study, we examined the effects on chondrogenesis of these transgenes when delivered in combination. Cultures of bone marrow-derived MSCs were infected with 2.5 x 10(2) or 2.5 x 10(3) viral particles/cell of each adenoviral vector individually, or in combination, seeded into aggregates, and cultured for 3 weeks in a defined serum-free medium. Levels of transgene product in the medium were initially high, approximately 100 ng/mL TGF-beta1, 120 ng/mL BMP-2, and 80 ng/mL IGF-1 at day 3, and declined thereafter. We found that co-expression of IGF-1 and TGF-beta1, BMP-2, or both at low doses resulted in larger aggregates, higher levels of glycosaminoglycan synthesis, stronger staining for proteoglycans and collagen type II and X, and greater expression of cartilage-specific marker genes than with either transgene alone. Gene-induced chondrogenesis of MSCs using multiple genes that act synergistically may enable the administration of reduced viral doses in vivo and could be of considerable benefit for the development of cell-based therapies for cartilage repair.


MiR-374b promotes osteogenic differentiation of MSCs by degrading PTEN and promoting fracture healing.
To investigate whether miR-374b can promote the differentiation of MSCs into osteoblasts by mediating PTEN, thus promoting fracture healing. Primary cultured mouse mesenchymal stem cells were obtained for following experiments. Flow cytometry was used to determine the expression of MSCs surface antigens to identify the purity. Alizarin red staining was used to detect whether MSCs could differentiate into osteoblasts. QRT-PCR was used to detect the expression of osteogenic marker genes as well as miR-374b and PTEN in bone marrow-derived MSCs from fractured mice model. ALP activity detection kit was used to detect ALP activity in cells. Changes of osteogenic proteins in cells were evaluated by Western blot. Bioinformatics methods were used to predict the binding sites between miR-374 and target genes. Luciferase reporter assay was used to confirm whether miR-374 could bind to the target gene. Under normal culture, MSCs grew into a long fusiform shape on the 4th day. After induced in the osteogenic induction medium for seven days, calcified nodules appeared. The results of the detection of MSCs surface antigen markers showed that CD90 was 99.12%, and CD45 was 0.23%. Alizarin red staining showed that MSCs possess the ability to differentiate into osteogenic. The expression level of MiR-374b and PTEN increased significantly in the early stage of fracture in mice, but no significant difference was observed at a later stage. After overexpression of miR-374b, the cell ALP activity, the expression of osteogenesis-related genes and osteogenesis-related proteins was significantly increased, while after knocking out miR-374b, the opposite result was observed. The result of luciferase reporting assay showed that miR-374b can bind to PTEN. As mentioned above, overexpression of miR-374b resulted in upregulation of osteogenic-related genes and proteins, while over-expression of both PTEN and miR-374b could partly reverse the outcomes. miR-374b can promote osteogenic differentiation of MSCs by degrading PTEN, thereby promoting fracture healing.


Structural and functional studies of hemoglobin Poissy alpha 2 beta 2(56) (D7) Gly----Arg and 86 (F2) Ala----Pro.
Hemoglobin Poissy alpha 2 beta 2(56) (D7) Gly----Arg and 86 (F2) Ala----Pro, is a new variant of the beta chain with two substitutions within the second exon of the corresponding gene. The electrophoretic mobilities are identical to those of Hb Hamadan alpha 2 beta 2(56) (D7) Gly----Arg as is the fingerprint of the tryptic hydrolysate of the two abnormal beta chains. The second substitution beta 86 Ala----Pro was detected by high-pressure liquid chromatography. Hb Poissy has a threefold increase in oxygen affinity with low Hill coefficient and diminished Bohr effect, which are restored to normal upon addition of 2,3-bisphosphoglycerate. Since the functional properties of Hb Hamadan (beta 56 Gly----Arg) have been described as normal, the abnormal function of Hb Poissy may be attributed to the beta 86 (F2) Ala----Pro substitution. Hb Poissy exhibits a mild instability and a greater reactivity of the thiol groups of the beta 93 (F9) Cys residues in the deoxy form than does Hb A. The oxidation rate of Hb Poissy is biphasic indicating a large inequivalence between the alpha and beta hemes. Thereafter NMR studies demonstrated that the beta 86 Ala----Pro substitution produces a displacement of the F helix closer to the heme plane and a large increase in the dynamic fluctuations of the tertiary structure on the proximal side of the beta hemes. These results lead to the conclusion that the beta 86 Ala----Pro substitution produces a destabilization of the F helix extending downwards to the FG corner and altering both the beta hemes and the alpha 1 beta 2 contacts.


Effect of low-density lipoprotein apheresis on circulating endothelial progenitor cells in familial hypercholesterolemia.
Long-term treatment with low-density lipoprotein (LDL) apheresis (LA) has been shown to reduce the incidence of cardiovascular events in patients affected by familial hypercholesterolemia (FH). Data from experimental studies suggest that circulating endothelial progenitor cells (EPCs) can repair the vascular lesions caused by atherosclerosis. Since a reduction of these cells has been demonstrated to predict atherosclerosis progression, the aim of this study was to verify whether LA can increase the percentage of EPCs. In 15 patients affected by FH periodically treated with LA, the percentage of EPCs was determined before and after performing LA, and compared with the values of 15 control subjects and 15 hypercholesterolemic patients treated with statins. Significant differences were found in FH patients between the pre-apheresis percentages of CD34+/KDR+, defined as EPCs by a wide consensus of opinion, and the values found 24 h after the procedures (0.00868 +/- 0.003 vs. 0.01009 +/- 0.002%, p < 0.005). Instead, the percentages of CD34+/KDR+/CD133+, considered as an immature subset of EPCs, remained substantially unchanged. However, a significant reduction in the percentage of EPCs was observed in both patient groups as compared to the controls, at all the assessment times. In the short-term LA seems to stimulate mobilization of CD34+/KDR+ cells. Hypercholesterolemic patients show a lower percentage of EPCs than controls. There were no differences in the EPCs percentages between the 2 patients groups, despite the fact that LDL cholesterol levels were higher in the group undergoing LA.


Potential Role of IL-37 in Atopic Dermatitis.
Interleukin 37 (IL-37) is a recently discovered member of the IL-1 cytokine family that appears to have anti-inflammatory and immunosuppressive effects in various diseases. IL-37 acts as a dual-function cytokine, exerting its effect extracellularly by forming a complex with the receptors IL-18 α (IL-18Rα) and IL-1R8 and transmitting anti-inflammatory signals, as well as intracellularly by interacting with Smad3, entering the nucleus, and inhibiting the transcription of pro-inflammatory genes. Consequently, IL-37 is linked to IL-18, which plays a role in the pathogenesis of atopic dermatitis (AD), consistent with our studies. Some isoforms of IL-37 are expressed by keratinocytes, monocytes, and other skin immune cells. IL-37 has been found to modulate the skewed T helper 2 (Th2) inflammation that is fundamental to the pathogenesis of AD. This review provides an up-to-date summary of the function of IL-37 in modulating the immune system and analyses its potential role in the pathogenesis of AD. Moreover, it speculates on IL-37's hypothetical value as a therapeutic target in the treatment of AD.


Characterization of the prodynorphin and proenkephalin neuropeptide systems in rat hippocampus.
Opioid peptides derived from prodynorphin were localized immunocytochemically to dentate granule cells and mossy fibers of the rat hippocampus with antisera against dynorphin A(1-17) and dynorphin B. Extracts of microdissected hippocampal regions were resolved by reverse phase and molecular exclusion chromatography to identify the molecular forms of the dynorphin A immunoreactivity and to quantify regional contents. Results demonstrated that the relative concentration of dynorphin A within each dissected region of hippocampus agreed well with the distribution of dynorphin A detected by immunocytochemical methods. Immunostaining of proenkephalin-derived opioid peptides, [Leu5]enkephalin and bovine adrenal medullary peptide-22P, was concentrated in cell bodies of the entorhinal cortex, nerve fibers in the perforant pathway, and terminals in the outer molecular layer of the dentate gyrus. Light immunostaining of granule cells and mossy fibers with these antisera was also found. The relative concentration of [Leu5]enkephalin immunoreactivity in each microdissected region of the hippocampus also agreed well with the distribution of [Leu5]enkephalin immunostaining. Chromatography of hippocampal regional extracts demonstrated that the immunoreactivity measured was due to the presence of authentic [Leu5]enkephalin. The probable neurotransmitter function of both [Leu5]enkephalin and dynorphin A was shown by their calcium-dependent release after in vitro depolarization of hippocampal tissue. The reported presence of beta-endorphin in hippocampus was not verified. Comparison of the hippocampal distribution and content of prodynorphin and proenkephalin-derived opioids suggests that separate populations of neurons containing these two peptide families form distinct neurotransmitter systems of roughly equal concentration.


NEWTON-2 Cisternal (Nimodipine Microparticles to Enhance Recovery While Reducing Toxicity After Subarachnoid Hemorrhage): A Phase 2, Multicenter, Randomized, Open-Label Safety Study of Intracisternal EG-1962 in Aneurysmal Subarachnoid Hemorrhage.
A sustained release microparticle formulation of nimodipine (EG-1962) was developed for treatment of patients with aneurysmal subarachnoid hemorrhage (aSAH). To assess safety, tolerability, and pharmacokinetics of intracisternal EG-1962 in an open-label, randomized, phase 2 study of up to 12 subjects. Subjects were World Federation of Neurological Surgeons grades 1 to 2, modified Fisher grades 2 to 4, and underwent aneurysm clipping within 48 h of aSAH. EG-1962, containing 600 mg nimodipine, was administered into the basal cisterns. Outcome on the extended Glasgow Outcome Scale (eGOS), pharmacokinetics, delayed cerebral ischemia and infarction, rescue therapy, and safety were evaluated. The study was halted when a phase 3 study of intraventricular EG-1962 stopped because that study was unlikely to meet its primary endpoint. Six subjects were randomized (5 EG-1962 and 1 oral nimodipine). After 90-d follow-up, favorable outcome on the eGOS occurred in 1 of 5 EG-1962 and in the single oral nimodipine patient. Four EG-1962 and the oral nimodipine subject had angiographic vasospasm. One EG-1962 subject had delayed cerebral ischemia, and all subjects with angiographic vasospasm received rescue therapy except 1 EG-1962 patient. One subject treated with EG-1962 developed right internal carotid and middle cerebral artery narrowing 5 mo after placement of EG-1962, leading to occlusion and cerebral infarction. Pharmacokinetics showed similar plasma concentrations of nimodipine in both groups. Angiographic vasospasm and unfavorable clinical outcome still occurred after placement of EG-1962. Internal carotid artery narrowing and occlusion after placement of EG-1962 in the basal cisterns has not been reported.


A new method of chronic and recurrent seminal vesiculitis treatment.
To investigate a new method and its effect on the procedure of dilating the ejaculatory duct and flushing the seminal vesicle with an F9 seminal vesicle scope in patients with chronic and recurrent seminal vesiculitis. Twenty-six patients with a diagnosis based to signs, laboratory detection, digital rectal examination, and transrectal ultrasonography were involved in present study. The patients underwent a surgical procedure of dilating the ejaculatory duct and flushing the seminal vesicles with an F9 seminal vesicle endoscope. All patients were followed for 3 months to 1 year after treatment. There were significant reductions in symptoms, signs, white blood cell and red blood cell counts on microscopic examination, seminal vesicles size, improvement of inner walls echo in transrectal ultrasonography, and semen culture positive rate. Moreover, all patients showed improvement. The present study provides a new transurethral seminal tract endoscopic technique with seminal vesicle scope through the normal anatomic tract to treat patients with chronic seminal vesiculitis. It proved to be easily conducted with minimized complications. Further investigations are needed to confirm our results.


Recent Emergence of Anaplasma phagocytophilum in Ontario, Canada: Early Serological and Entomological Indicators.
Human granulocytic anaplasmosis (HGA), caused by the bacteria Anaplasma phagocytophilum, is transmitted to humans by blacklegged ticks (Ixodes scapularis) in eastern North America. To assess the emergence of A. phagocytophilum in Ontario, we analyzed patient serological and clinical data in combination with pathogen detection in blacklegged ticks from 2011 to 2017. Our sample population included all patients who had Anaplasma serological testing ordered by their physicians (n = 851). Eighty-three patients (10.8%) were A. phagocytophilum seropositive (IgG titers ≥ 1:64) and 686 (89.2%) were seronegative (IgG titers < 1:64). Applying published surveillance case definitions, we classified zero as confirmed, five as probable, and 78 as suspected cases. The percentage of seropositive patients remained generally stable at 13.6%. Seropositive patients were most often adult females, 40-59 years of age, and reported nonspecific signs and symptoms, such as fatigue, headache, and fever. Higher seropositivity rates (≥ 1.5 patients per 100,000 population) occurred in eastern and northwestern Ontario. The percentage of A. phagocytophilum-positive blacklegged ticks, through passive and active surveillance, was 0.4 and 1.1%, respectively, and increased over time. Serological and entomological indicators of A. phagocytophilum activity increased in areas of the province with established blacklegged tick populations. The risk of HGA is presently low in Ontario; however, further research is required to document the epidemiology of HGA in the province. To minimize the impact of HGA emergence in Ontario, increased awareness and education of the public and health-care providers is recommended, with consideration to making HGA a reportable infection in Ontario.


The integrase promoter and T'I terminator in bacteriophages lambda and 434.
The lambda integrase promoter PI lies immediately downstream from a terminator T'I. The PI promoter is activated by cII protein during lysogenization. The function of T'I is unknown. A deletion (trp-lambda 29), whose fusion point lies within one stem of T'1 retains cII-activable promoter function but shows little if any transcription termination in vivo. The nucleotide sequence of PI is identical in lambda and the related phage 434. However, the T'I sequences of the two phages, though located in the same position, are not detectably homologous. When restriction fragments carrying the promoter-terminator segments from each of these phages were inserted into the trp operon, both responded identically to cII activation, and both caused termination.


Biliary tract cancer and genomic alterations in homologous recombinant deficiency: exploiting synthetic lethality with PARP inhibitors.
Biliary tract cancers (BTC) are a group of rare, chemoresistant solid tumor malignancies that arise from the bile ducts. BTC are typically associated with poor outcomes. Most patients present with advanced disease, where treatment is palliative with platinum based cytotoxic therapy. Response to chemotherapy is variable with limited duration of response. A subset of patients that will receive durable and meaningful responses to platinum-based chemotherapy is deemed to be platinum sensitive. The availability and implementation of next-generation sequencing allowed genomic profiling of BTC, which have identified potential targetable somatic genetic aberrations, which include kinases (FGFR, BRAF, ALK, ERBB2), oncogenes (IDH1/2, CCND1) and tumor suppressor genes, including germline or somatic mutations involved in DNA damage response (DDR) genes. These genes include, but are not limited to: ATM, ATR, BRCA1/2, RAD51, PALB2, PTEN, FANC, NBN, EMSY, MRE11, ARID1A. In BTC, alterations in DDR genes are identified in up to 20% of patients, with a higher proportion identified in those with extrahepatic cholangiocarcinoma. Patients harboring mutations exhibit varying patterns of clinical behavior and response to therapy. The presence of these mutations typically predicts for susceptibility to DNA damaging chemotherapy, such as platinum agents.


Depicting the anatomy of the gyral white matter: ubi sumus? quo vadimus?
A cerebral gyrus is made up of an external layer of folded cortex and an inner core of white matter. The architecture of the core has specific features that make it distinct from the white matter of the deep brain regions. Limited externally by the grey matter that covers the top of the gyrus and the neighbouring sulci, this gyral white matter is made up of a mix of fibre populations with multiple directions and destinations. The presence of densely packed fibres with multiple crossings, the proximity to the cortex and the existence of inter-regional and inter-individual variations make the task of depicting this microanatomy extremely challenging. The topic is, however, of paramount relevance for both fundamental and applied neurosciences. This fibre colocalization is crucial for the functional role of each cerebral region and is key to clinical manifestations in cases of parenchymal damage. As track tracing, imaging and dissection are based on different biological or physical principles, it is natural for their results to sometimes be different, but they are often complementary. As the amount of available information increases, it becomes fragmented due to the multiplicity of methods, target phenomena and studied species. In this scoping review, we present the key concepts and map the primary sources of evidence regarding identifying the fibre pathways that compose the gyral white matter, enabling the discussion of avenues for future research. The general pattern in which these pathways are distributed in the gyral white matter was detailed, and the main variations as a function of brain topography were explained and illustrated with typical examples.


Cell type and maturation stage-dependent polymorphism of N-linked oligosaccharides on murine lymphocytes and lymphoma cells.
We analyzed the overall structures of N-linked oligosaccharides on glycoproteins of various murine lymphocytic and lymphoma cells employing a newly developed method which was performed on high-performance liquid chromatography after derivatization of oligosaccharides with 2-aminopyridine. A total of 15 types of bi, tri- and tetra-antennary N-acetyllactosamine-type oligosaccharides with or without fucose and oligomannose-type oligosaccharides were identified on these cells in variable amounts depending on the type and maturation stage of the cells. It was found that all murine lymphocytic cells carry N-acetyllactosamine-type oligosaccharides with the additional alpha-linked galactose residue on the non-reducing ends. Thymocytes had exceptionally large amounts of oligosaccharides with one or even two alpha-galactose residues per molecule. In contrast, peripheral resting T cells possessed those oligosaccharides only in a small amount, although the cells produced more the oligosaccharides after stimulation with Con A. Two thymoma lines such as BW 5147 and EL-4 and one B cell lymphoma line WEHI231 contained relatively large amount of oligosaccharides with alpha-galactose residues. Significant change of the molar ratio of component carbohydrates by cell activation was observed also in oligommanose-type oligosaccharides which were few in resting T cells but were markedly increased in Con A activated cells. Molar ratio of triantennary oligosaccharides in total N-acetyllactosamine type oligosaccharides was high in thymocytes and low in resting T cells, but was increased in T cells after Con A activation. It was also very high in WEHI 231 B cell lymphoma. Although BW 5147 and EL-4 thymoma did not contain tri-antennary oligosaccharides in high proportion, they carried larger tetra-antennary oligosaccharides with an N-acetyllactosamine repeating unit in definitive amounts. It is suggested from these results that overall structures of oligosaccharides on cell surface proteins of lymphocytes are finely controlled with link to cell differentiation, activation and transformation.


When Differing Perspectives Between Health Care Providers and Parents Lead to "Communication Crises": A Conceptual Framework to Support Prevention and Navigation in the Pediatric Hospital Setting.
The communication experience and therapeutic relationships between parents and health care providers (HCPs) impacts the quality of patient care. A guiding "communication crisis" description was created to encourage study participants to describe difficult communication encounters between parents and HCPs where their perspectives regarding the recommended patient care conflicted (ie, parent refusal of a lumbar puncture), which created barriers to the provision of optimal care and the development of therapeutic relationships in the pediatric hospital setting. The purpose of this research was to highlight factors that may contribute to communication crises through the characterization of these circumstances. Participants were multidisciplinary HCPs and parents (n = 37) with firsthand experience regarding communication crises. Data were collected through focus groups (7), semistructured interviews (2), and a verification focus group where open-ended questions regarding participants' experiences were used. Data were analyzed by using a constructivist grounded theory approach. Three themes and 11 subthemes (communication crisis risk factors) were identified: (1) health care team factors (communication skills, care processes, and interprofessional communication), (2) family and/or parent factors (language or cultural barriers, mental health conditions, socioeconomic factors, and beliefs), (3) patient factors (acute condition, unclear diagnosis, unstable condition, and medical complexity). A core theory emerged: parent trust in their HCP significantly impacts the therapeutic relationship and can mitigate communication crises despite the presence of risk factors. We highlight factors that may be predisposing to communication crises in pediatric hospital settings. Awareness of these factors can support timely identification and implementation of relationship care and foster the establishment of trusting relationships.


Analysis of Catecholamine and Their Metabolites in Mice Brain by Liquid Chromatography-Mass Spectrometry Using Sulfonated Mixed-mode Copolymer Column.
In this study, a simultaneous assay for catecholamines and their metabolites in the brain was established using liquid chromatography-mass spectrometry (LC-MS). To achieve complete separation, a cation-exchange/reversed-phase mixed-mode copolymer resin column containing 0.81 wt% sulfo groups was used for the simultaneous LC-MS assay. The analyzed catecholamines were dopamine (DA), norepinephrine (NE), and epinephrine (E), while the metabolites lacking amino groups were 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 3-methoxy-4-hydroxyphenylglycol (MHPG). The metabolites were separated and detected using LC-MS, on columns with and without sulfo groups. However, we could not achieve adequate separation of catecholamines on both columns using a gradient elution of 0 - 50 (v/v)% methanol containing 0.1 (v/v)% formic acid (FA). When volatile ion-pairing reagents were added to the mobile phase, they improved the retention and detection of catecholamines on the sulfonated mixed-mode column. Under optimized elution conditions, which involved a linear gradient elution of water containing 0.1 (v/v)% FA to 50 (v/v)% acetonitrile in 50 mM ammonium formate at 40°C and a 0.20 mL/min rate, all six target molecules were simultaneously detected within 25 min, when using negative mode LC-MS on a sulfonated mixed-mode column. The limits of detection (LODs) for DA, NE, E, DOPCA, HVA, and MHPG were determined to be 20.7, 12.6, 74.6, 1110, 18.7, and 3196 nM, respectively. Moreover, the established LC-MS assay allowed the detection of endogenous DA, NE, and HVA, in normal mouse brain samples at concentrations higher than 20, 9, and 4 pmol/mg, respectively.