Food protein-induced enterocolitis syndrome is an important differential diagnosis of vomiting in pediatric emergency.
The aim of this study was to clarify the clinical characteristics and management of infants with suspected acute food protein-induced enterocolitis syndrome (FPIES) who presented to the pediatric emergency department (ED) before and after the guidelines were published. This was a retrospective cohort study. We classified the infants as "acute FPIES-like symptoms" who were younger than 12 months of age and visited the pediatric ED of the National Center for Child Health and Development due to vomiting 1-4 h after food ingestion without any causative disease, such as infection. The medical records of those infants in 2015 and 2021 were reviewed. We used the nonparametric Mann-Whitney U test to compare two groups for continuous variables, whereas chi-squared or Fisher's exact tests were used for nominal variables. The number of infants with acute FPIES-like symptoms was 15 (13%) in 2015 and 14 (15%) in 2021. The trigger foods were cow's milk or dairy products in half of the infants, and five as a result of ingestion of hen's eggs in 2021, compared to zero cases in 2015. Five in 2015 and 12 in 2021 required examination at the ED. Three in 2015 and six in 2021 met the diagnostic criteria for acute FPIES in the international consensus guidelines. The emergency physicians did not record at least four minor criteria for acute FPIES in seven in 2015 and five in 2021. No infants (0%) in 2015 and two (14%) in 2021 were referred to the allergy department by an emergency physician. Acute FPIES should be considered one of the differential diagnoses of vomiting, and pediatric medical staff should be aware of FPIES diagnostic criteria and appropriately refer suspected cases to a specialist.


The Effect of Respiratory Viral Infections on Breakthrough Hemolysis in Patients with Paroxysmal Nocturnal Hemoglobinuria.
Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by hemolysis and thrombosis and is associated with significant morbidity and mortality. Although complement inhibitors have significantly changed the outcomes in PNH patients, breakthrough hemolysis (BTH) may still occur as a response to stress factors such as pregnancy, surgery, and infections. Despite the well-described association between bacterial infections and hemolysis in PNH patients, little is known about the effect of respiratory viruses on triggering hemolytic episodes. This is the first study, to our knowledge, addressing this question. We retrospectively analyzed 34 patients with PNH disease between 2016 and 2018, who were on eculizumab treatment and who presented with respiratory symptoms and were subsequently tested for 10 respiratory viruses (influenza A, influenza B, parainfluenza, respiratory syncytial virus, adenovirus, rhinovirus, and human metapneumovirus). NTS+ patients had higher inflammatory markers, with the majority requiring antibiotics. Acute hemolysis, along with a significant drop in hemoglobin, was noted in the NTS+ group, with three of them requiring a top-up transfusion and two requiring an extra dose of eculizumab. Furthermore, the time from the last eculizumab dose was longer in the NTS+ patients who had BTH, than those who did not. Our data indicate that respiratory virus infections pose a significant risk for BTH in PNH patients on complement inhibitor treatment, underlining the need for regular screening and close monitoring of patients with respiratory symptoms. Furthermore, it implies a higher risk for patients who are not established on complement inhibitors, suggesting the necessity for greater vigilance in these patients.


COMPARING THE EFFECTIVENESS OF CONCENTRATE OF ALL PROTHROMBIN COMPLEX FACTORS WITH RECOMBINANT HUMAN COAGULATION FACTOR VIIa IN THE TREATMENT OF BLEEDING AFTER CARDIAC SURGERY.
Aim: The authors evaluated the effectiveness of treatment with recombinant human coagulation factor VIIa and concentrate of all prothrombin complex factors in patients with massive postoperative bleeding that could not be controlled with traditional therapy. Materials and Methods: In the period from 2020 to 2021, recombinant human coagulation factor VIIa was administered to 18 patients after cardiac surgery (group I), while the concentrate of all prothrombin complex factors was administered to 16 patients postoperatively (group II). During this period, 647 patients were operated on. The patients had normal coagulation screening tests (APTT, INR, TT, fibrinogen level, and PLT level) before surgery. Mean blood loss before and after administration of eptacog alfa and the total prothrombin complex concentrate was assessed. The mean dose of eptacog alfa was 30.95 mcg/kg b.w., and the total prothrombin complex factor concentrate dose was 14.17 mcg/kg b.w. After transfusion with red blood cell concentrate, fresh frozen plasma, and platelet concentrate, in the absence of improvement in the dynamics of postoperative drainage, it was decided to include recombinant human coagulation factor VIIa or a concentrate of all prothrombin complex factors in the treatment. Results: After administration of recombinant human coagulation factor VIIa at a dose of 30.95 mcg/kg b.w., bleeding stopped in 12 patients, but the remaining 6 patients required reoperation due to persistently high drainage. The decision to perform a rethoracotomy was made by a team of cardiothoracic surgeons and anesthesiologists, taking into account the dynamics of drainage (bleeding) and the hemodynamic stability of the patient. After the administration of concentrate of all prothrombin complex factors at a dose of 14.17 U/kg b.w., bleeding stopped in 12 patients. Four patients required reoperation due to persistent bleeding. Conclusions: Treatment with recombinant human coagulation factor VIIa and concentrate of all prothrombin complex factors is effective and safe for cardiac surgery patients.


Expression of the metastasis-associated MTA1 protein and its relationship to deacetylation of the histone H4 in esophageal squamous cell carcinomas.
Metastasis-associated protein MTA1 and histone deacetylase form a protein complex with histone deacetylase activity that plays an important role in histone deacetylation, alteration of chromatin structure and transcriptional control. The precise role of the MTA1 protein in the malignant progression of human cancers remains unknown, however, especially its overexpression and relationship with histone acetylation/deacetylation in experimental and clinical tumors. The expression levels of MTA1 protein and the acetylation levels of histone H4 were examined in 70 cases of surgically resected esophageal squamous cell carcinomas, using immunohistochemistry. The intensities of immunostaining of MTA1 protein and acetylated histone H4 in carcinoma tissues (Ca) were compared to normal epithelium (N) contained in the same section. Thirty of 70 cases (42.9%) displayed overexpression of MTA1 protein (N < Ca). Cancers overexpressing MTA1 protein invaded deeper into the esophageal wall (p < 0.005) and showed significantly higher degrees of lymph node metastasis (p < 0.01), higher pathological stage, more lymphatic involvement and poorer prognosis (p < 0.05) than the remaining cases. The acetylation levels of histone H4 inversely correlated to the depth of cancer invasion and pathological stage (p < 0.05), and the patients with higher level of histone H4 acetylation had a better prognosis (p < 0.05). Furthermore, immunostaining patterns of MTA1 and acetylated histone H4 were inversely correlated (p < 0.001), demonstrating the relationship of deacetylation of histone H4 in MTA1-overexpressing carcinomas. In conclusion, the data suggest that the overexpression of MTA1 protein and acetylation level of histone H4 protein, both of which are closely related, might be useful predictors of malignant potential of esophageal squamous cell carcinomas. Thus, strategies involving inhibition of MTA1 function as well as inhibition of histone deacetylation could be novel approaches for the treatment of esophageal squamous cell carcinomas.


[Clinical usefulness of inhibin assays in gynecology and obstetrics].
The main source of inhibin B in women is the growing follicle granulosa cells, while inhibin A is mainly produced by the corpus luteum and the placenta. In infertile women submitted to therapies of assisted reproduction, inhibin B has shown to be useful to predict a poor ovulatory response, though it has not yet overcome the performance of other markers. In the pre-natal screening of the Down syndrome, inhibin A has been repeatedly confirmed as useful in the second trimester and has also started to be considered in the first trimester test battery. Besides the two applications above, the dosage of total inhibin may contribute to the identification of cases of autoimmune ovarian insufficiency. Total inhibin may also be an auxiliary marker in the diagnosis of ovarian epithelial tumors, while the amount of inhibin B helps in the diagnosis of granulosa cells tumors. The use of inhibin A may be extended to the evaluation of pregnant women with risk of abortion, with a history of repeated abortion, with increased risk of pre-eclampsia, or even in the first days of follow-up of hydatiform mole post-emptying. All those applications are still under study, but with a real possibility of helping to extend the diagnostic spectrum of inhibin dosage in gynecology and obstetrics.


Antibacterial activity of oral antibiotics against community-acquired respiratory pathogens from three European countries.
Antimicrobial resistance is universally recognized as a major problem. A European resistance survey was established to monitor the activity of widely used oral antibiotics against common respiratory tract pathogens. Studies were conducted in Italy, Spain and Austria to monitor resistance patterns among respiratory Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pyogenes, Staphylococcus aureus and Klebsiella pneumoniae to amoxicillin, co-amoxiclav, penicillin, cefaclor, cefadroxil, cefalexin, cefprozil, cefuroxime, cefixime, ceftibuten, cefpodoxime, clarithromycin and azithromycin (the antibiotics tested varying slightly from country to country). Minimum inhibitory concentrations were determined using the NCCLS-recommended broth microdilution method. Among the antibiotics tested, cefpodoxime, an oral cephalosporin, was remarkably active against the major respiratory pathogens in all three countries. Cefpodoxime was more potent than cefaclor, cefixime and ceftibuten against pneumococci, especially against strains with decreased sensitivity to penicillin, and more active than cefaclor and cefuroxime against Gram-negative respiratory pathogens. Pneumococci and staphylococci displayed a very high level of in vitro macrolide resistance. These data indicate that cefpodoxime represents an appropriate choice in the treatment of community-acquired respiratory tract infection in the three countries surveyed.


Effects of different perfusates on functional parameters of isolated perfused dog kidneys.
The isolated perfused canine kidney has been established as a valid model for conducting both renal physiology and transplantation research. This model is of particular importance for developing new strategies to improve graft function after renal transplantation. In the present study, a newly developed method using isolated haemoperfused porcine kidneys was adapted for use in canine kidneys. In contrast to haemoperfusion, synthetic perfusion media can be standardized and can prevent the initiation of blood-mediated reperfusion reactions. Thus, an additional aim was to determine whether blood could be replaced by synthetic cell-free perfusion solutions. Canine kidneys (n = 30) were harvested from donors euthanized in veterinary practices for causes unrelated to the present study. The kidneys were isolated and perfused with autologous blood or cell-free synthetic electrolyte buffer (Tyrode solution). During perfusion, we monitored renal perfusate flow (RPF), glomerular filtration rate (GFR), electrolyte and glucose reabsorption, oxygen consumption and urine concentration. Changes in perfusion medium did not affect the RPF. In contrast, GFR, urine concentration and oxygen consumption were significantly higher, whereas fractional excretion of sodium and glucose were significantly lower in blood- than in Tyrode-perfused kidneys. This system offers a simple model for studying whole-organ functional alterations after acute renal ischaemia. Renal function indicators were below values reported during in vivo physiological conditions. These functions were better conserved when kidneys were perfused with autologous blood than with Tyrode.


Patient perceptions about nutrition and skin health: a survey study characterizing patient opinions and information resources.
Numerous studies in the clinical literature have explored the link between nutrition and skin physiology. However, it is unclear whether patients visit their dermatologists with knowledge of these studies, and unknown where they obtain their skin health information. We characterized patient perceptions surrounding nutrition and skin, including what patients identified as aggravating and alleviating foods and their information sources. We administered a questionnaire to 409 participants attending University of California (UC) Davis Dermatology and Pacific Skin Institute in Sacramento. This survey assessed their perception on the influence of nutrition. We stratified responses by diseases. Of the 409 respondents, 83% believed that nutrition affects skin health. Respondents with healthy skin were not more likely to agree than those with skin conditions in general (P=0.34). Those with skin conditions also more likely received their information from reputable sources, defined as physicians and scientific journals (P=0.02). Additionally, respondents who disagreed were more likely informed by reputable sources (P=0.002), but when online blogs were included as reputable, this relationship was less significant (P=0.046). As online resources become more accessible, it is important for providers to know about changing patient perspectives. Our findings may help improve how dermatologists counsel patients about nutrition.


Outcomes of Nonoperative Management of Uncomplicated Appendicitis.
Nonoperative management (NOM) of uncomplicated pediatric appendicitis has promise but remains poorly studied. NOM may lead to an increase in resource utilization. Our objective was to investigate the trends in NOM for uncomplicated appendicitis and study the relevant clinical outcomes including subsequent appendectomy, complications, and resource utilization. Retrospective analysis of administrative data from 45 US pediatric hospitals. Patients <19 years of age presenting to the emergency department (ED) with appendicitis between 2010 and 2016 were studied. NOM was defined by an ED visit for uncomplicated appendicitis treated with antibiotics and the absence of appendectomy at the index encounter. The main outcomes included trends in NOM among children with uncomplicated appendicitis and frequency of subsequent diagnostic imaging, ED visits, hospitalizations, and appendectomy during 12-month follow-up. 99 001 children with appendicitis were identified, with a median age of 10.9 years. Sixty-six percent were diagnosed with nonperforated appendicitis, of which 4190 (6%) were managed nonoperatively. An increasing number of nonoperative cases were observed over 6 years (absolute difference, +20.4%). During the 12-month follow-up period, NOM patients were more likely to have the following: advanced imaging (+8.9% [95% confidence interval (CI) 7.6% to 10.3%]), ED visits (+11.2% [95% CI 9.3% to 13.2%]), and hospitalizations (+43.7% [95% CI 41.7% to 45.8%]). Among patients managed nonoperatively, 46% had a subsequent appendectomy. A significant increase in NOM of nonperforated appendicitis was observed over 6 years. Patients with NOM had more subsequent ED visits and hospitalizations compared with those managed operatively at the index visit. A substantial proportion of patients initially managed nonoperatively eventually had an appendectomy.


Target cells for the polychlorinated biphenyl metabolite 4,4'-bis(methylsulfonyl)-2,2',5,5'-tetrachlorobiphenyl in lung and kidney.
Light microscope autoradiography was used to determine the cellular localization of the polychlorinated biphenyl metabolite 4,4'-bis([14C] methylsulfonyl)-2,2',5-5'-tetrachlorobiphenyl ([3H]TCB) in the lung and kidney of mice and rats. Microautoradiograms prepared from thaw-mounted freeze sections showed that the radioactivity in the lung was localized in the bronchiolar lumen and epithelium. In methacrylate sections (74-82% of radioactivity extracted), a highly selective labeling was registered in the apical cytoplasm of the Clara cells. A pronounced labeling was present also in certain goblet-like cells containing periodic acid-Schiff-positive granules. Gel permeation chromatography and density gradient centrifugation showed that 80-95% of the radioactivity in lung lavage fluid was bound to a specific protein previously characterized in rat and mouse lung cytosol. The protein appeared to be enriched in the lavage fluid, as compared to lung cytosol. These data suggest that [3H]TCB binds to a protein residing in the Clara and goblet-like cells and that the labeled TCB-protein complex is subsequently secreted into the airway lumen. As shown by microautoradiography, the radioactivity in the kidney was confined to a restricted portion of the nephron, predominantly to the apical region of the proximal tubular cells.


Zebrafish behavioural assays of translational relevance for the study of psychiatric disease.
Understanding the pathogenesis of the complex behavioural disorders that constitute psychiatric disease is a major challenge for biomedical research. Assays in rodents have contributed significantly to our understanding of the neural basis of behavioural disorders and continue to be one of the main focuses for the development of novel therapeutics. Now, owing to their genetic tractability and optical transparency (allowing in vivo imaging of circuit function) and the rapid expansion of genetic tools, zebrafish are becoming increasingly popular for behavioural genetic research. The increased development of behavioural assays in zebrafish raises the possibility of exploiting the advantages of this system to identify molecular mechanisms contributing to behavioural phenotypes associated with psychiatric disorders as well as potential therapeutics. This mini-review describes behavioural paradigms in zebrafish that can be used to address endophenotypes associated with psychiatric disease. The content reflects the interests of the author and covers tests of cognitive functions, response choice and inhibition, social interaction and executive function.


Azapirones for Attention Deficit Hyperactivity Disorder: A Systematic Review.
No meta-analysis has evaluated azapirones (serotonin1A receptor partial agonists) as anxiolytics for attention deficit hyperactivity disorder (ADHD). Randomized controlled trials (RCTs) and single-arm trials published before October 27, 2015 were retrieved from major healthcare databases and clinical trial registries. Relative risk and 95% confidence intervals were calculated. 5 RCTs (n=429) and 3 single-arm studies (n=70) were identified. 3 RCTs compared buspirone vs. methylphenidate in children/adolescents, one buspirone patches vs. placebo patches in children/adolescents, and one atomoxetine plus buspirone vs. atomoxetine vs. placebo in adults. The single-arm studies were buspirone trails in children/adolescents. All-cause discontinuation rates and adverse events did not differ between pooled buspirone and methylphenidate groups. No other meta-analyses of buspirone efficacy and safety vs. comparators were conducted due to insufficient data. 2 RCTs found no significant differences in parent and teacher ADHD-Rating Scale total scores between buspirone and methylphenidate, while one reported that methylphenidate improved parent and teacher ADHD-RS total scores vs. buspirone. It remains unclear whether buspirone use has benefit for ADHD patients and therefore further evidence is needed for better clinical use of buspirone in patients with ADHD.


Structures and relative free energies of partially folded states of proteins.
The ability of proteins to fold to well defined compact structures is one of the most remarkable examples of the effect of natural selection on biological molecules. To understand their properties, including the stability, the mechanism of folding, and the possibilities of misfolding and association, it is necessary to know the protein free energy landscape. We use NMR data as restraints in a Monte Carlo sampling procedure to determine the ensemble of structures populated by human alpha-lactalbumin in the presence of increasing concentrations of urea. The ensembles of structures that represent the partially folded states of the protein show that two structural cores, corresponding to portions of the alpha and beta domains of the native protein, are preserved even when the native-like interactions that define their existence are substantially weakened. Analysis of the network of residual contacts reveals the presence of a complex interface region between the two structural cores and indicates that the development of specific interactions within this interface is the key step in achieving the native structure. The relative probabilities of the conformations determined from the NMR data are used to construct a coarse-grained free energy landscape for alpha-lactalbumin in the absence of urea. The form of the landscape, together with the existence of distinct cores, supports the concept that robustness and modularity are the properties that make possible the folding of complex proteins.


Acquired resistance to BRAF inhibitors mediated by a RAF kinase switch in melanoma can be overcome by cotargeting MEK and IGF-1R/PI3K.
BRAF is an attractive target for melanoma drug development. However, resistance to BRAF inhibitors is a significant clinical challenge. We describe a model of resistance to BRAF inhibitors developed by chronic treatment of BRAF(V)⁶⁰⁰(E) melanoma cells with the BRAF inhibitor SB-590885; these cells are cross-resistant to other BRAF-selective inhibitors. Resistance involves flexible switching among the three RAF isoforms, underscoring the ability of melanoma cells to adapt to pharmacological challenges. IGF-1R/PI3K signaling was enhanced in resistant melanomas, and combined treatment with IGF-1R/PI3K and MEK inhibitors induced death of BRAF inhibitor-resistant cells. Increased IGF-1R and pAKT levels in a post-relapse human tumor sample are consistent with a role for IGF-1R/PI3K-dependent survival in the development of resistance to BRAF inhibitors.


Management of Co-Morbidity of Depression and Chronic Non-Communicable Diseases in Rwanda.
Chronic non-communicable diseases (NCDs) are a major global health problem of the 21 stcentury. They are now the world's leading cause of disease burden and high mortality. An even more alarming health problem is when depression coexists with chronic NCDs, as is frequently the case. Management of this co-morbidity with collaborative care has become a global topic of interest, with the World Health Organization (WHO) recommending implementation of collaborative care for this purpose. The study investigated existing protocols and/or interventions for managing this co-morbidity in Rwandan district hospitals. The study used an action research design involving a research team of 14 health care professionals to collaboratively identify existing protocols or interventions for managing co-morbidity of depression and NCDs in Rwanda. Focus group discussion using a structured interview guide was used to collect qualitative data, followed by qualitative content analysis using inductive approach. We found no particular protocols or interventions in place to manage the co-morbidity of depression and chronic NCDs. Depression and chronic NCDs were found to be treated separately, in separate health care settings and by different health professionals. The findings revealed a gap in management of co-morbid depression and chronic NCDs in Rwanda district hospitals. We recommend that health care providers follow the WHO collaborative care advisory for better quality care and better patient improvement in management of this co-morbidity.


Finite element analysis of Bi-condylar Tibial Plateau fractures to assess the effect of coronal splits.
Bi-condylar tibial plateau fractures are demanding to treat due to the complex geometry and the articular comminution. The presence of a coronal fracture line plays a crucial role in the fixation strategy. Disregarding this fracture line in previous biomechanical studies and established fracture classifications resulted in a lack of detailed knowledge regarding the influence of medial-posterior fragments on implant load sharings. This study aimed to evaluate the effects of coronal splits on stress distributions within the implants using the finite element analysis (FEA). FE models with (Fracture C) and without the coronal split (Fracture H) were developed and validated in order to assess stress distributions within the implant components. Comparing FE outcomes with biomechanical experiments indicated that both fracture models were well validated. FE evaluations demonstrated that the coronal split caused destabilization of the medial tibia, as well as a shift in the peak-stress areas from the middle part of the plate to the proximal section, and a 61% increase in the maximum stress of the kick-stand screw. Therefore, FE models based on clinically-relevant fracture morphologies can provide a reliable tool to assess implant failures as well as to compare different fracture fixation techniques.


Binding pattern analysis and structural insight into the inhibition mechanism of Sterol 24-C methyltransferase by docking and molecular dynamics approach.
Sterol 24-C methyltransferase (SMT) plays a major role during the production of steroids, especially in the biosynthesis of ergosterol, which is the major membrane sterol in leishmania parasite, and the etiological basis of leishmaniasis. Mechanism-based inactivators bind irreversibly to SMT and interfere with its activity to provide leads for the design of antileishmanial inhibitors. In this study, computational methods are used for studying enzyme-inhibitor interactions. fifty-seven mechanism-based inactivators are docked using 3 docking/scoring approaches (FRED, GoldScore, and ChemScore). A consensus is generated from the results of different scoring functions which are also validated with already reported experimental values. The most active compound thus obtained is subjected to molecular dynamics simulation of length 20 ns. Stability of simulation is analyzed through root-mean-square deviation, beta factor (B-factor), and radius of gyration (Rg). Hydrogen bonds and their involvement in the structural stability of the enzyme are evaluated through radial distribution function. Newly developed application of axial frequency distribution that determines three-particle correlation on frequency distributions before and after simulation has provided a clear evidence for the movement of the inhibitor into active pocket of the enzyme. Results yielded strong interaction between enzyme and the inhibitor throughout the simulation. Binding of the inhibitor with enzyme has stabilized the enzyme structure; thus, the inhibitor has the potential to become a lead compound.


Physiological and symptom determinants of exercise performance in patients with chronic airway obstruction.
To evaluate the physiological and symptom determinants of exercise performance (EP) as measured by a 6-min walking test (6MWD), Watt(max), and peak oxygen consumption (VO2 ml/min/kg), 105 patients with chronic airway obstruction (CAO) [50 chronic obstructive pulmonary disease (COPD): 44 men, aged 63+/-7 years, forced expiratory volume in 1 sec (FEV1) forced vital capacity (FVC)(-1)% 54+/-13; and 55 asthmatic: 23 men, aged 55+/-10 years, FEV1 FVC(-1) % 65+/-10] underwent evaluation of 6MWD, symptom limited cyclo-ergometer exercise test, spirometry, respiratory muscle function, arterial blood gases and sensation of dyspnoea [using the Borg scale, Visual Analogue Scale (VAS) and Baseline Dyspnoea Index (BDI)]. A hierarchical method of analysis identified the residual volume (RV), total lung capacity (TLC)(-1) ratio, BDI and the patient's age as the strongest and most consistent correlates of EP (r2 = 0.14-0.21). The correlation between EP and its various determinants was not influenced by diagnosis. The relationship between breathlessness and EP was different between men and women: at any given level of exercise, women were more breathless than men. In multivariate analyses that contained both RV TLC(-1) and BDI, the RV TLC(-1) ratio was the strongest correlate of EP, although the BDI remained a significant covariate. Overall, age was the major determinant of EP but inclusion of the RV TLC(-1) ratio and the BDI into the model explained a further 9-15% of the variance in EP. These three covariates together explained 26-34% of the variance between patients. We conclude that in stable CAO patients, the prediction of exercise capacity by anthropometric, demographic, clinical and physiological variables is likely to be low. Age, pulmonary hyperinflation and dyspnoea are the strongest and most consistent correlates of impaired exercise performance. Airways obstruction, measured during expiration using FEV1, does not appear to be a predictor of physiological impairment. These results underline the importance of performing exercise evaluation in CAO patients.


Myricetin improves endurance capacity and mitochondrial density by activating SIRT1 and PGC-1α.
Robust mitochondrial respiration provides energy to support physical performance and physiological well-being, whereas mitochondrial malfunction is associated with various pathologies and reduced longevity. In the current study, we tested whether myricetin, a natural flavonol with diverse biological activities, may impact mitochondrial function and longevity. The mice were orally administered myricetin (50 mg/kg/day) for 3 weeks. Myricetin significantly potentiated aerobic capacity in mice, as evidenced by their increased running time and distance. The elevated mitochondrial function was associated with induction of genes for oxidative phosphorylation and mitochondrial biogenesis in metabolically active tissues. Importantly, myricetin treatment led to decreased PGC-1α acetylation through SIRT1 activation. Furthermore, myricetin significantly improved the healthspan and lifespan of wild-type, but not Sir-2.1-deficient, C. elegans. These results demonstrate that myricetin enhances mitochondrial activity, possibly by activating PGC-1α and SIRT1, to improve physical endurance, strongly suggesting myricetin as a mitochondria-activating agent.


Is grid therapy useful for all tumors and every grid block design?
Grid therapy is a treatment technique that has been introduced for patients with advanced bulky tumors. The purpose of this study is to investigate the effect of the radiation sensitivity of the tumors and the design of the grid blocks on the clinical response of grid therapy. The Monte Carlo simulation technique is used to determine the dose distribution through a grid block that was used for a Varian 2100C linear accelerator. From the simulated dose profiles, the therapeutic ratio (TR) and the equivalent uniform dose (EUD) for different types of tumors with respect to their radiation sensitivities were calculated. These calculations were performed using the linear quadratic (LQ) and the Hug-Kellerer (H-K) models. The results of these calculations have been validated by comparison with the clinical responses of 232 patients from different publications, who were treated with grid therapy. These published results for different tumor types were used to examine the correlation between tumor radiosensitivity and the clinical response of grid therapy. Moreover, the influence of grid design on their clinical responses was investigated by using Monte Carlo simulations of grid blocks with different hole diameters and different center-to-center spacing. The results of the theoretical models and clinical data indicated higher clinical responses for the grid therapy on the patients with more radioresistant tumors. The differences between TR values for radioresistant cells and radiosensitive cells at 20 Gy and 10 Gy doses were up to 50% and 30%, respectively. Interestingly, the differences between the TR values with LQ model and H-K model were less than 4%. Moreover, the results from the Monte Carlo studies showed that grid blocks with a hole diameters of 1.0 cm and 1.25 cm may lead to about 19% higher TR relative to the grids with hole diameters smaller than 1.0 cm or larger than 1.25 cm (with 95% confidence interval). In sum-mary, the results of this study indicate that grid therapy is more effective for tumors with radioresistant characteristics than radiosensitive tumors.


Estimation of zinc concentration of parotid saliva by flameless atomic absorption spectrophotometry in normal subjects and in patients with idiopathic hypogeusia.
The concentraiton of zinc in parotid saliva was estimated by flameless atomic absorption spectrophotometry. As small a sample as 5 to 10 mul of whole paratid saliva can be analyzed quickly and reliably, without special sample handling and with littleinterference from other ions normally found in saliva. Parotid salivary zinc concentration in 34 subjects with normal taste acuity was 51 plus or minus 14 ppb(parts per billion)(mean plus or minus 1 S.D.); parotid salivary zinc concentration in 47 subjectswith idiopathic hypogeusia was 10 plus or minus 6 ppb (mean plus or minus 1 S.D.) which is significantly lower than that in normal subjects (p lessthan 0.001). This technique is a usual method by which to evaluate parotid salivary zinc, particulary inrelationship to problems of taste, appetite, and nutrition.


Improving healthcare services using web based platform for management of medical case studies.
The paper presents a web based platform for management of medical cases, support for healthcare specialists in taking the best clinical decision. Research has been oriented mostly on multimedia data management, classification algorithms for querying, retrieving and processing different medical data types (text and images). The medical case studies can be accessed by healthcare specialists and by students as anonymous case studies providing trust and confidentiality in Internet virtual environment. The MIDAS platform develops an intelligent framework to manage sets of medical data (text, static or dynamic images), in order to optimize the diagnosis and the decision process, which will reduce the medical errors and will increase the quality of medical act. MIDAS is an integrated project working on medical information retrieval from heterogeneous, distributed medical multimedia database.


Experimental infection with a Thai reassortant swine influenza virus of pandemic H1N1 origin induced disease.
Following the emergence of the pandemic H1N1 influenza A virus in 2009 in humans, this novel virus spread into the swine population. Pigs represent a potential host for this virus and can serve as a mixing vessel for genetic mutations of the influenza virus. Reassortant viruses eventually emerged from the 2009 pandemic and were reported in swine populations worldwide including Thailand. As a result of the discovery of this emergent disease, pathogenesis studies of this novel virus were conducted in order that future disease protection and control measures in swine and human populations could be enacted. The pandemic H1N1 2009 virus (pH1N1) and its reassortant virus (rH1N1) isolated from pigs in Thailand were inoculated into 2 separate cohorts of 9, 3-week-old pigs. Cohorts were consisted of one group experimentally infected with pH1N1 and one group with rH1N1. A negative control group consisting of 3 pigs was also included. Clinical signs, viral shedding and pathological lesions were investigated and compared. Later, 3 pigs from viral inoculated groups and 1 pig from the control group were necropsied at 2, 4, and 12 days post inoculation (DPI). The results indicated that pigs infected with both viruses demonstrated typical flu-like clinical signs and histopathological lesions of varying severity. Influenza infected-pigs of both groups had mild to moderate pulmonary signs on 1-4 DPI. Interestingly, pigs in both groups demonstrated viral RNA detection in the nasal swabs until the end of the experiment (12 DPI). The present study demonstrated that both the pH1N1 and rH1N1 influenza viruses, isolated from naturally infected pigs, induced acute respiratory disease in experimentally inoculated nursery pigs. Although animals in the rH1N1-infected cohort demonstrated more severe clinical signs, had higher numbers of pigs shedding the virus, were noted to have increased histopathological severity of lung lesions and increased viral antigen in lung tissue, the findings were not statistically significant in comparison with the pH1N1-infected group. Interestingly, viral genetic material of both viruses could be detected from the nasal swabs until the end of the experiment. Similar to other swine influenza viruses, the clinical signs and pathological lesions in both rH1N1 and pH1N1 were limited to the respiratory tract.


Unified patch clamp protocol for the characterization of Pannexin 1 channels in isolated cells and acute brain slices.
In the central nervous system, Pannexin 1 (Panx1) channels are implicated in a variety of physiological and pathological conditions. One of the prerequisites to enlighten the role of Panx1 is the development and standardization of reliable methods. Here, we address the applicability of voltage clamp protocols to identify Panx1 channel mediated currents in neurons of acutely dissected brain slices. We improved an established protocol and report on a modified paradigm that robustly evokes Panx1 channel currents. Crucial advances are the use of physiologic ion gradient conditions and a preconditioning step of depolarizing membrane potential ramps of long duration. This new paradigm provides significant impact on membrane current generation at hypo- and depolarized holding potential steps post voltage ramp preconditioning in heterologous expression systems and primary hippocampal CA1 neurons of mouse brain slices in vitro. Finally, we demonstrate that under these conditions the analysis of tail currents elicited by repolarization of the cells from preconditioning holding potential depolarization permits an independent method to isolate Panx1 mediated channel activity. In summary, this study provides a comprehensive methodological improvement in the biophysical analysis of Panx1 channels with a particular focus on investigations under physiological conditions in complex tissues.


Schistosoma mansoni genome project: an update.
A schistosome genome project was initiated by the World Health Organization in 1994 with the notion that the best prospects for identifying new targets for drugs, vaccines, and diagnostic development lie in schistosome gene discovery, development of chromosome maps, whole genome sequencing and genome analysis. Schistosoma mansoni has a haploid genome of 270 Mb contained on 8 pairs of chromosomes. It is estimated that the S. mansoni genome contains between 15000 and 25000 genes. There are approximately 16689 ESTs obtained from diverse libraries representing different developmental stages of S. mansoni, deposited in the NCBI EST database. More than half of the deposited sequences correspond to genes of unknown function. Approximately 40-50% of the sequences form unique clusters, suggesting that approximately 20-25% of the total schistosome genes have been discovered. Efforts to develop low resolution chromosome maps are in progress. There is a genome sequencing program underway that will provide 3X sequence coverage of the S. mansoni genome that will result in approximately 95% gene discovery. The genomics era has provided the resources to usher in the era of functional genomics that will involve microarrays to focus on specific metabolic pathways, proteomics to identify relevant proteins and protein-protein interactions to understand critical parasite pathways. Functional genomics is expected to accelerate the development of control and treatment strategies for schistosomiasis.


S-wave predominance of epicardial electrograms during atrial fibrillation in humans: indirect evidence for a role of the thin subepicardial layer.
The purpose of this study was to characterize the morphology of fibrillation electrograms in patients in order to provide insight into the underlying electropathologic substrate of atrial fibrillation (AF). Electrograms recorded during AF show a high degree of spatiotemporal variation. AF was induced by rapid atrial pacing in 25 patients undergoing cardiac surgery. A unipolar mapping array of 244 electrodes was positioned on the free wall of the right atrium to record multiple epicardial fibrillation electrograms. Local anisotropy in conduction and epicardial wavefront curvature during AF were determined by fitting the best quadratic surface on the activation times of rectangular areas of 3 x 3 electrodes. During AF, unipolar epicardial electrograms revealed a clear predominance of S waves. The average RS difference during type I and II AF was -0.15 +/- 0.08 and -0.22 +/- 0.08. During type III AF, the predominance of S waves was less prominent (-0.07 +/- 0.05; P < .005). In all types of AF, the degree of anisotropy in conduction was remarkably low (anisotropy ratio: 1.24 +/- 0.09), and no clear directional effect on the relative amplitude of R and S waves was found. There was a weak relationship between local curvature of wavefronts and RS difference (r = 0.23; P < .01). Computer simulations showed that the negative RS difference could result from transmural activation in an epicardial to endocardial direction. The clear predominance of S waves in epicardial fibrillation electrograms is not due to anisotropy and can only be partly explained by a high curvature of fibrillation waves. Predominant epicardial to endocardial activation seems to be important in producing rS electrograms on the epicardium. This finding provides indirect evidence that the thin epicardial layer of atrial myocardium plays an important role in propagation of fibrillation waves.


Dopamine Transporter Is a Master Regulator of Dopaminergic Neural Network Connectivity.
Dopaminergic neurons of the substantia nigra pars compacta (SNC) and ventral tegmental area (VTA) exhibit spontaneous firing activity. The dopaminergic neurons in these regions have been shown to exhibit differential sensitivity to neuronal loss and psychostimulants targeting dopamine transporter. However, it remains unclear whether these regional differences scale beyond individual neuronal activity to regional neuronal networks. Here, we used live-cell calcium imaging to show that network connectivity greatly differs between SNC and VTA regions with higher incidence of hub-like neurons in the VTA. Specifically, the frequency of hub-like neurons was significantly lower in SNC than in the adjacent VTA, consistent with the interpretation of a lower network resilience to SNC neuronal loss. We tested this hypothesis, in DAT-cre/loxP-GCaMP6f mice of either sex, when activity of an individual dopaminergic neuron is suppressed, through whole-cell patch clamp electrophysiology, in either SNC or VTA networks. Neuronal loss in the SNC increased network clustering, whereas the larger number of hub-neurons in the VTA overcompensated by decreasing network clustering in the VTA. We further show that network properties are regulatable via a dopamine transporter but not a D2 receptor dependent mechanism. Our results demonstrate novel regulatory mechanisms of functional network topology in dopaminergic brain regions.SIGNIFICANCE STATEMENT In this work, we begin to untangle the differences in complex network properties between the substantia nigra pars compacta (SNC) and VTA, that may underlie differential sensitivity between regions. The methods and analysis employed provide a springboard for investigations of network topology in multiple deep brain structures and disorders.


Mucoepidermoid carcinoma of the lacrimal gland. An oxyphilic variant.
Unlike their major salivary gland counterparts, mucoepidermoid carcinomas (MECs) of the lacrimal gland are extraordinarily rare. We report a case of an MEC of the lacrimal gland in which the majority of cells were oxyphilic. The patient, a 38-year-old woman, had been asymptomatic when an enlarged right lacrimal gland was detected. Histologically, the tumor was composed primarily of oxyphilic cells but focally exhibited characteristics diagnostic of MEC. To our knowledge, only two oxyphilic variants of MEC have been reported, and occurrence in the lacrimal gland has not previously been documented. The existence of this lesion is significant because it shares histologic features with oxyphilic adenoma (oncocytoma), a more common and less aggressive neoplasm of the orbit.


Temporal variation in the prevalence and species richness of Campylobacter spp. in a prairie watershed impacted by urban and agricultural mixed inputs.
Campylobacter spp. are a substantial cause of gastroenteritis worldwide. Human infection can result from ingestion of contaminated food or water from a variety of sources, including the consumption of fresh produce that is contaminated with the pathogen via the use of contaminated irrigation water. Using molecular methods, we investigated the occurrence of Campylobacter in the Qu'Appelle River watershed, an important source of irrigation water for vegetable producers in southern Saskatchewan, Canada. Water samples were collected from 7 sampling sites from April to September 2009 (145 samples), and from 5 sampling sites from May to October 2013 (116 samples). Campylobacter was detected in 57% and 16% of the samples collected in 2009 and 2013, respectively. Campylobacter detection was highest in May and June for both sampling years. In 2009, the predominant species were Campylobacter lari and Campylobacter jejuni, with prevalences of 84% and 41%, respectively. Other Campylobacter spp. were detected less frequently. Only C. lari was detected in 2013. The results in 2009 demonstrate the species richness of Campylobacter in water sources within the watershed. The occurrence of Campylobacter in the study area also underscores the importance of monitoring irrigation water used to irrigate fresh produce from a public health prospective.


[Immunological effect of systemically administered allopurinol in experimental autoimmune uveitis].
Allopurinol shows beneficial effects in the systemic treatment of lens-induced uveitis. This is believed to be due to the reduction of oxidative tissue damage via a dose-dependent free radical scavenging ability and an immunomodulating effect. The purpose of this study was to investigate the immunological effects in experimental autoimmune uveitis after systemic treatment with allopurinol (AL) and steroids (STER). 31 male Lewis rats were immunized with crude retinal extract, Freund's Adjuvans and pertussis toxin. The rats were divided into four groups: healthy rats (BASIS, n = 3), experimental autoimmune-uveitis without therapy (EAU, n = 9), 50 mg/kg bw. allopurinol i.v. (ALSYS, n = 9), and 7.5 mg/kg bw. methylprednisolone i.v. (STSYS, n = 10). ALSYS and STSYS received five intravenous injections during the 2 weeks immunization period. The rats' sera were tested against Western Blots (WB) of electrophoretic separations of retinal proteins. Based on digital image analysis, an analysis of discriminance was performed. The multivariate analysis of discriminance revealed a significance difference between the WBs of ALSYS and STSYS (p < 0.01) compared to EAU without therapy. The number and intensity of peaks in WBs were strongly reduced in the ALSYS group compared to EAU. AL revealed a strong immunomodulating effect in the treatment of experimental autoimmune uveitis that is markedly stronger than that of steroids. Together with the antioxidative effect of allopurinol known from previous studies, this drug could be a new promising therapeutic approach in the treatment of uveitis.


The relationship between hippocampal volume and declarative memory in a population of combat veterans with and without PTSD.
Both reduced hippocampal volume and cognitive alterations have been found in posttraumatic stress disorder (PTSD). The purpose of this article was to examine the relationship between hippocampal volume, combat exposure, symptom severity, and memory performance in a sample of combat veterans with and without a history of PTSD. Subjects were 33 male veteran volunteers (16 PTSD+, 17 PTSD-) who underwent an MRI and neuropsychological testing with the California Verbal Learning Test (CVLT), a measure of declarative memory. Relationships between hippocampal volume (i.e., right + left hippocampal volume/whole brain volume) and performance on the CVLT were determined using partial correlational analysis controlled for age and Wechsler Adult Intelligence Scale, Third Edition (WAIS-III) vocabulary scores. Percent hippocampal volume for the entire sample was positively associated with several aspects of memory performance as reflected by the CVLT. In the PTSD+ group, CVLT performance was negatively correlated with lifetime, but not current CAPS symptoms. CVLT performance appears to be strongly correlated with hippocampal volume in a group of trauma survivors with and without PTSD. Insofar as CVLT performance in the PTSD group was negatively associated with worst episode, but not to current PTSD symptoms, memory performance in combat veterans may reflect some aspect of risk related to the magnitude of the psychological response to trauma, rather than current symptoms that may be interfering with cognitive performance. It will be of interest to study cognitive abilities that may relate to the likelihood of specific PTSD symptoms and to track changes in CVLT performance and hippocampal volume over time in persons with and without a history of trauma exposure.


Evaluation of a commercially available radioimmunoassay and enzyme immunoassay for the analysis of progesterone and estradiol and the comparison of two extraction efficiency methods.
The measurement of progesterone (P4) and estradiol (E2) is essential for monitoring reproductive cycles and can aid in diagnosing the cause of poor reproductive performance in dairy cattle. Readily available, reproducible, accurate, non-radioactive assays are needed for the assessment of P4 and E2 in bovine serum. The gold standard for hormone assessment, radioimmunoassay (RIA), was compared with enzyme-linked immunoassay (EIA). Serum collected from various points in the estrous cycle was extracted with radiolabeled P4 (ie, 3H-P4; HE) and without 3H-P4 (CE) before being used in the assay. For the assessment of P4, there is a great degree of correlation between the RIA and EIA (adjusted R-square = 0.95; Pearson correlation coefficient (PCC) = 0.98, P < 0.001). A difference between the RIA and EIA methods was not detected for E2 concentrations (P = 0.16), but the correlation between techniques was poor (adjusted R-squared = 0.73; PCC = 0.87, P = 0.002). There was no difference in the serum extraction efficiency as measured with 3H-P4 as opposed to without (P = 0.94). The two methods for the measurement of serum extraction efficiency were highly correlated (adjusted R-square = 0.83; PCC = 0.92, P < 0.001). The concordance correlation coefficient (CCC) showed an excellent agreement between RIA and EIA for P4 determination (0.89) and between HE and CE methods (0.90). Although the 95% limits of agreement of the Bland-Altman plots encompassed 89% (8/9) and 92% (12/13) of the differences between methods for P4 quantification and extraction respectively, the CCC indicated an excellent agreement among them. The CCC between RIA and EIA for E2 quantification was 0.68 which corresponds with a fair agreement; however, the 95% limits of agreement of the Bland-Altman plot encompassed 100% (9/9) of differences between methods. The EIA and CE methods are comparable alternatives to the RIA and HE methods, respectively and can be used to quantify P4 and E2 for bovine serum.


[Is bone microstructure different between osteopenic and osteoporotic patients with femoral neck fracture?].
This study aims to investigate whether bone microstructure is different between osteopenic and osteoporotic patients with femoral neck fracture. Fourteen patients (5 males, 9 females) with proximal femoral fracture following low energy trauma were included in the study. Patients with secondary osteoporosis and who received antiosteoporotic treatment prior to trauma were excluded. The patients were divided into two groups based on the bone mineral density measurement, including osteopenic group (n=7, mean age 69 years; range 63-74 years) and osteoporotic group (n=7, mean age 74.1 years; range 67-78 years). Cortical bone specimens for biopsy obtained from the patients who underwent endoprosthesis during partial hip arthroplasty and were evaluated using light microscope. Mean cortical bone thickness was 3622.14 μM in osteopenic group, and 2323.14 μM in osteoporotic group (p<0.005). However, light microscope revealed similar characteristics for both groups. Although a significant difference was found between the groups in terms of cortical thickness, bone microstructure showed similar characteristics in osteopenic and osteoporotic patients with low energy femoral neck fracture.


Mimicking natural cell environments: design, fabrication and application of bio-chemical gradients on polymeric biomaterial substrates.
Gradients of biomolecules on synthetic, solid substrates can efficiently mimic the natural, graded variation of properties of the extracellular matrix (ECM). Such gradients represent accessible study platforms for the understanding of cellular activities, and they also provide functional supports for tissue engineering (TE). This review describes the most relevant methods to produce 2-dimensional (2D) as well as 3-dimensional (3D) gradient supports for cell manipulations, and also addresses the response of cells from different origins when seeded on these constructs. The fabrication strategies summarized encompass the combination of polymer and surface chemistries, micro- and nano-engineering construction strategies and biotechnological approaches. This multidisciplinary scheme has enabled the design and realization of diverse, synthetic supports as cellular environments, spanning from the first gradient self-assembled monolayer (SAM) to multilayers, and hybrid constructs mimicking the complexity of natural tissue environments. The standing challenge is bringing these advances in the fabrication of supports to a dynamic functioning in space and time, towards the successful imitation of the most complex bio-chemical system ever studied: our body.


Clinical significance of high-sensitivity C-reactive protein in cardiovascular disease.
Large-scale prospective cohort studies consistently demonstrate a strong, independent relationship between high-sensitivity (hs) C-reactive protein (CRP) and incident cardiovascular events, with a magnitude of effect similar to, or larger than, that of blood pressure and lipid levels. As a biomarker of inflammation, hsCRP levels also predict incident Type 2 diabetes and modify the risk associated with the metabolic syndrome. Recent work further demonstrates that the addition of hsCRP to information provided by traditional risk factors improves risk classification, particularly for individuals otherwise considered to be at intermediate risk. Although there remains no direct evidence that lowering hsCRP lowers vascular risk, optimal clinical outcomes have been observed in statin trials among patients who not only reduced low-density lipoprotein cholesterol below 1.8 mmol/l (70 mg/dl), but who also reduced hsCRP below 2 mg/l. In addition to statins, CRP levels are lowered by diet, exercise and smoking cessation, all of which are known to lower vascular event rates. Whether or not CRP represents a causal agent in atherosclerosis is controversial and an area in need of further research. However, this controversy does not diminish the clinical utility of hsCRP as a biomarker of risk that, if appropriately used in clinical practice, can substantially improve clinical care.


Evolutionary contribution to coexistence of competitors in microbial food webs.
The theory of species coexistence is a key concept in ecology that has received much attention. The role of rapid evolution for determining species coexistence is still poorly understood although evolutionary change on ecological time-scales has the potential to change almost any ecological process. The influence of evolution on coexistence can be especially pronounced in microbial communities where organisms often have large population sizes and short generation times. Previous work on coexistence has assumed that traits involved in resource use and species interactions are constant or change very slowly in terms of ecological time-scales. However, recent work suggests that these traits can evolve rapidly. Nevertheless, the importance of rapid evolution to coexistence has not been tested experimentally. Here, we show how rapid evolution alters the frequency of two bacterial competitors over time when grown together with specialist consumers (bacteriophages), a generalist consumer (protozoan) and all in combination. We find that consumers facilitate coexistence in a manner consistent with classic ecological theory. However, through disentangling the relative contributions of ecology (changes in consumer abundance) and evolution (changes in traits mediating species interactions) on the frequency of the two competitors over time, we find differences between the consumer types and combinations. Overall, our results indicate that the influence of evolution on species coexistence strongly depends on the traits and species interactions considered.


Time for Using Machine Learning for Dose Guidance in Titration of People With Type 2 Diabetes? A Systematic Review of Basal Insulin Dose Guidance.
Real-world studies of people with type 2 diabetes (T2D) have shown insufficient dose adjustment during basal insulin titration in clinical practice leading to suboptimal treatment. Thus, 60% of people with T2D treated with insulin do not reach glycemic targets. This emphasizes a need for methods supporting efficient and individualized basal insulin titration of people with T2D. However, no systematic review of basal insulin dose guidance for people with T2D has been found. To provide an overview of basal insulin dose guidance methods that support titration of people with T2D and categorize these methods by characteristics, effect, and user experience. The review was conducted according to the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) guidelines. Studies about basal insulin dose guidance, including adults with T2D on basal insulin analogs published before September 7, 2022, were included. Joanna Briggs Institute critical appraisal checklists were applied to assess risk of bias. In total, 35 studies were included, and three categories of dose guidance were identified: paper-based titration algorithms, telehealth solutions, and mathematical models. Heterogeneous reporting of glycemic outcomes challenged comparison of effect between the three categories. Few studies assessed user experience. Studies mainly used titration algorithms to titrate basal insulin as telehealth or in paper format, except for studies using mathematical models. A numerically larger proportion of participants seemed to reach target using telehealth solutions compared to paper-based titration algorithms. Exploring capabilities of machine learning may provide insights that could pioneer future research while focusing on holistic development.


Five new pedigrees with inherited RUNX1 mutations causing familial platelet disorder with propensity to myeloid malignancy.
Familial platelet disorder with propensity to myeloid malignancy (FPD/AML) is an autosomal dominant syndrome characterized by platelet abnormalities and a predisposition to myelodysplasia (MDS) and/or acute myeloid leukemia (AML). The disorder, caused by inherited mutations in RUNX1, is uncommon with only 14 pedigrees reported. We screened 10 families with a history of more than one first degree relative with MDS/AML for inherited mutations in RUNX1. Germ- line RUNX1 mutations were identified in 5 pedigrees with a 3:2 predominance of N-terminal mutations. Several affected members had normal platelet counts or platelet function, features not previously reported in FPD/AML. The median incidence of MDS/AML among carriers of RUNX1 mutation was 35%. Individual treatments varied but included hematopoietic stem cell transplantation from siblings before recognition of the inherited leukemogenic mutation. Transplantation was associated with a high incidence of complications including early relapse, failure of engraftment, and posttransplantation lymphoproliferative disorder. Given the small size of modern families and the clinical heterogeneity of this syndrome, the diagnosis of FPD/AML could be easily overlooked and may be more prevalent than previously recognized. Therefore, it would appear prudent to screen young patients with MDS/AML for RUNX1 mutation, before consideration of sibling hematopoietic stem cell transplantation.


The role of cannabinoid transmission in emotional memory formation: implications for addiction and schizophrenia.
Emerging evidence from both basic and clinical research demonstrates an important role for endocannabinoid (ECB) signaling in the processing of emotionally salient information, learning, and memory. Cannabinoid transmission within neural circuits involved in emotional processing has been shown to modulate the acquisition, recall, and extinction of emotionally salient memories and importantly, can strongly modulate the emotional salience of incoming sensory information. Two neural regions in particular, the medial prefrontal cortex (PFC) and the basolateral nucleus of the amygdala (BLA), play important roles in emotional regulation and contain high levels of cannabinoid receptors. Furthermore, both regions show profound abnormalities in neuropsychiatric disorders such as addiction and schizophrenia. Considerable evidence has demonstrated that cannabinoid transmission functionally interacts with dopamine (DA), a neurotransmitter system that is of exceptional importance for both addictive behaviors and the neuropsychopathology of disorders like schizophrenia. Research in our laboratory has focused on how cannabinoid transmission both within and extrinsic to the mesolimbic DA system, including the BLA → mPFC circuitry, can modulate both rewarding and aversive emotional information. In this review, we will summarize clinical and basic neuroscience research demonstrating the importance of cannabinoid signaling within this neural circuitry. In particular, evidence will be reviewed emphasizing the importance of cannabinoid signaling within the BLA → mPFC circuitry in the context of emotional salience processing, memory formation and memory-related plasticity. We propose that aberrant states of hyper or hypoactive ECB signaling within the amygdala-prefrontal cortical circuit may lead to dysregulation of mesocorticolimbic DA transmission controlling the processing of emotionally salient information. These disturbances may in turn lead to emotional processing, learning, and memory abnormalities related to various neuropsychiatric disorders, including addiction and schizophrenia-related psychoses.


Avoiding the labrum in hip arthroscopy.
Iatrogenic joint damage is a potential complication of any arthroscopic procedure. The anatomic constraints of the hip cause particular concern. The greatest risks are perforation of the acetabular labrum and scuffing of the articular surface. Careful attention to the details of the technique described can reduce the likelihood of this problem. With current technology, some occasional damage is unavoidable. However, all steps should be taken to minimize the likelihood and magnitude of such occurrences.


Long-term outcome of adjunctive celecoxib treatment after paclitaxel-eluting stent implantation for the complex coronary lesions: two-year clinical follow-up of COREA-TAXUS trial.
In the COREA-TAXUS trial ("Effect of Celecoxib On REstenosis after coronary Angioplasty with a TAXUS stent"), celecoxib reduced late luminal loss and adverse cardiac events at follow-up around 6 months. The objective of this study was to assess the long-term outcome of short-term adjunctive celecoxib treatment after paclitaxel-eluting stent implantation. This is a 2-year clinical follow-up of the COREA-TAXUS trial, an open-label randomized controlled study. A total 274 patients were randomized to receive or not receive celecoxib (400 mg before the intervention and 200 mg twice daily for 6 months after the procedure), and 271 underwent successful paclitaxel-eluting stent implantation. All patients were given aspirin (100 mg daily indefinitely) and clopidogrel (75 mg daily for at least 6 months). Among the 271 patients, 267 (98.5%) completed the 2-year clinical follow-up. From the previous follow-up to 2 years, there was no difference in the rate of adverse cardiac events between the celecoxib and control groups (1.6% versus 4.3%, P=0.27). Thus, at 2 years, the rate of adverse cardiac events was consistently lower in the celecoxib group (6.9% versus 19.7%, P=0.002). A significant reduction in need for target lesion revascularization was observed (6.2% versus 18.2%, P=0.003). The efficacy benefit in the celecoxib group was not undermined by an increased risk for cardiac death or myocardial infarction at 2 years (1.5% versus 1.4%). Six-month adjunctive celecoxib treatment after paclitaxel-eluting stent implantation was associated with durable long-term efficacy up to 2 years. However, the inconclusive evidence for the long-term safety of this treatment warrants caution. URL: http://www.clinicaltrials.gov. Unique identifier: NCT 00292721.


Technique and clinical applications of dual-energy contrast-enhanced digital mammography (CEDM) in breast cancer evaluation: a pictorial essay.
This pictorial essay illustrates an overview of the basic technique used in acquiring dual-energy contrast-enhanced digital mammography (CEDM) images and its potential clinical applications in regular practice. CEDM may be used as a low-cost alternative to magnetic resonance imaging (MRI), as a problem-solving tool in clinical practice and for therapeutic planning of breast cancer, which may include high-risk screening, dense breast evaluation, mammographically equivocal lesions, local staging, treatment response evaluation, and post treatment follow-up. We share our experience of CEDM at a tertiary care cancer hospital.


Neck scar perception after neck dissection in HPV-associated oropharyngeal squamous cell carcinoma.
While there has been an increased understanding of the impact of visible neck scars in some patients with certain diseases, this has not yet been explored in the HPV+ OPSCC population. To understand patients' perception of their scar and the impact on their quality of life (QOL) at least 6 months after neck dissection (ND) for HPV+ oropharyngeal squamous cell carcinoma (OPSCC). In this retrospective case-control study, patients who underwent primary transoral robotic surgery (TORS) and ND for HPV+ OPSCC between 2016 and 2021 at a single tertiary care center were identified. Data analysis was performed in January 2022. Dermatology Life Quality Index was modified (mDLQI) to assess patients' perceptions of their scars. The primary outcome was the mean mDLQI survey score with higher scores corresponding to worse perceptions. Three questions adapted from the Self-Consciousness Scale (SCS) were also included to assess awareness of appearance. All questions were scaled on a 0-3 Likert Scale. Tweedie generalized linear model was used to understand the relationship between mDLQI score and patient- and procedure-specific factors (including the three SCS survey questions). An additional exploratory logistic regression was performed to understand the risk factors for clinically significant mDLQI score change. A total of 67 patients (response rate 57 %) completed the survey with a mean mDLQI survey score of 0.84 (max 30). Although there was a statistically significant negative association between private insurance and mDLQI survey score (95 % Confidence Interval [CI]: -2.72 - -0.38), and a positive association between the SCS score and mDLQI survey score (95 % CI: 0.23-0.81) (p < 0.05), these variables were not found to be risk factors for a clinically significant difference in mDLQI on multivariable analysis. The majority of patients felt their neck scars did not interfere with their daily lives. Patient perceptions of neck scars were consistent despite differing patient characteristics and treatment regimens.


Carotid vascular control of ACTH secretion in lambs.
To test the hypothesis that changes in carotid vascular pressure influence ovine ACTH and corticosteroid secretion, we subjected 6- to 7-wk-old conscious lambs to bilateral carotid occlusion with or without prior vagosympathetic blockade. Reversible vagosympathetic blockade, produced by injection of 2% lidocaine onto the vagosympathetic trunks, inhibited the reflex changes in heart rate after intravenous injection of phenylephrine or during carotid occlusion. Vagosympathetic blockade by itself did not alter arterial pressure or heart rate but did increase circulating levels of ACTH and corticosteroids. This suggests that afferent fibers in the vagosympathetic trunks tonically inhibit the secretion of ACTH. Bilateral carotid occlusion did not change plasma ACTH or corticosteroid concentrations. Bilateral carotid occlusion plus prior vagosympathetic blockade stimulated increases in plasma ACTH and corticosteroid concentrations that were larger than responses to vagosympathetic blockade alone. The results demonstrate that changes in carotid vascular pressure influence ovine ACTH and corticosteroid secretion only during vagosympathetic blockade and therefore suggest that carotid vascular mechanoreceptors interact with other mechanoreceptors having vagosympathetic afferent fibers.


Food promotions and the cost of a healthy diet.
With approximately two in three UK adults overweight or obese, one in five living in poverty and our emergence from the Covid-19 pandemic with implications for employment and income status there is an urgent need to understand what it costs to eat healthily and the role that promotions can play in helping householders manage food budgets. The literature suggests that, in affluent countries, price promotions appear to increase consumer food purchases and are applied more frequently to less healthy products than their healthy counterparts. This review discusses the cost of a healthy diet, identifies the prevalence of promotions in both the supermarket setting generally and a typical shopping basket specifically, and discusses the barriers to affording a healthy diet. Given the current policy focus on the cost of living and population health emphasising the need for food shopping to represent health and value for money for better public health outcomes, this review contributes to the evidence base for retailers' and policymakers' consideration as policy solutions are sought to reduce population obesity levels, while ensuring the affordability and accessibility of nutritious food. It is important, given the shift in consumer purchasing behaviour to online shopping as a result of self-isolating or reticence to physically access stores in response to the Covid-19 pandemic, that retail food promotions are available irrespective of the chosen mode of shopping (in-store or online).


Acute Disorders of Consciousness in Pediatric Severe Sepsis and Organ Failure: Secondary Analysis of the Multicenter Phenotyping Sepsis-Induced Multiple Organ Failure Study.
Acute disorders of consciousness (DoC) in pediatric severe sepsis are associated with increased risk of morbidity and mortality. We sought to examine the frequency of and factors associated with DoC in children with sepsis-induced organ failure. Secondary analysis of the multicenter Phenotyping Sepsis-Induced Multiple Organ Failure Study (PHENOMS). Nine tertiary care PICUs in the United States. Children less than 18 years old admitted to a PICU with severe sepsis and at least one organ failure during a PICU stay. None. The primary outcome was frequency of DoC, defined as Glasgow Coma Scale (GCS) less than 12 in the absence of sedatives during an ICU stay, among children with severe sepsis and the following: single organ failure, nonphenotypeable multiple organ failure (MOF), MOF with one of the PHENOMS phenotypes (immunoparalysis-associated MOF [IPMOF], sequential liver failure-associated MOF, thrombocytopenia-associated MOF), or MOF with multiple phenotypes. A multivariable logistic regression analysis was performed to evaluate the association between clinical variables and organ failure groups with DoC. Of 401 children studied, 71 (18%) presented with DoC. Children presenting with DoC were older (median 8 vs 5 yr; p = 0.023), had increased hospital mortality (21% vs 10%; p = 0.011), and more frequently presented with both any MOF (93% vs 71%; p < 0.001) and macrophage activation syndrome (14% vs 4%; p = 0.004). Among children with any MOF, those presenting with DoC most frequently had nonphenotypeable MOF and IPMOF (52% and 34%, respectively). In the multivariable analysis, older age (odds ratio, 1.07; 95% CI, 1.01-1.12) and any MOF (3.22 [1.19-8.70]) were associated with DoC. One of every five children with severe sepsis and organ failure experienced acute DoC during their PICU stay. Preliminary findings suggest the need for prospective evaluation of DoC in children with sepsis and MOF.


Effects of progesterone and medroxyprogesterone on actin remodeling and neuronal spine formation.
Sex steroids are important regulators of neuronal cell morphology, and this is critical for gender differences in brain function and dysfunction. Neuronal morphology is controlled by multiprotein complexes including moesin (a member of the ezrin/radixin/moesin family), focal adhesion kinase (FAK), or the Wiskott-Aldrich syndrome protein-family verprolin homologous (WAVE1) protein, controlling dynamic remodeling of the cytoskeleton and cell membrane. We investigated the actions of natural progesterone (P) and of the synthetic progestin medroxyprogesterone acetate (MPA) on actin remodeling, focal adhesion complex formation, and actin branching in rat cortical neurons. Treatment with P and, to a lesser extent, MPA, increases the number and density of dendritic spines. P increases the phosphorylation of moesin, FAK, and WAVE1, and their redistribution toward cell membrane sites where spines are formed. Signaling to moesin is achieved by PR via a Gα/Gβ-dependent signaling to the small GTPase Ras homolog gene family, member A and its related kinase, Rho-associated kinase-2. In parallel, WAVE1 recruitment is triggered by a Gαi/Gβ-dependent signaling of PR to c-Src, FAK, and Rac1 GTPase. Rac1 recruits cyclin-dependent kinase-5, which phosphorylates WAVE1. Silencing of moesin, FAK, or WAVE1 abrogates the increase in dendritic spines induced by progesterone. In all applications, MPA is found to act similar to P, albeit with a lower efficacy. In conclusion, our findings indicate that the control of actin polymerization and branching and focal adhesion complex formation via moesin, FAK, and WAVE1 is a key function of progesterone receptor in neurons, which may be relevant for the regulation of dendritic spine turnover and neuronal plasticity.


[Isolation of enteroviruses from patients with clinical manifestations of poliomyelitis].
Examinations of 114 specimens (feces, nasopharyngeal washings, cerebrospinal fluid) from patients with clinical manifestations of poliomyelitis were carried out. Enteroviruses were isolated in 61.4% of the cases. Among the isolates, poliomyelitis viruses accounted for 49.1%, and Coxsackie B1 and ECHO 6 12.3%. Typing of the poliovirus isolates showed 17.9% to belong to type I, 7.1% to type II, and 75% to type III. Studies on intratypic differentiation revealed their antigenic similarity with the Mahoney, MEF-1, and Saukett strains, respectively. Among 24 paired blood sera tested for virus-neutralizing antibodies to wild poliovirus strains 21 demonstrated a 4-fold or higher rise of antibody titres to the above poliomyelitis viruses.


Plasminogen on the surfaces of fibrin clots prevents adhesion of leukocytes and platelets.
Although leukocytes and platelets adhere to fibrin with alacrity in vitro, these cells do not readily accumulate on the surfaces of fibrin clots in vivo. The difference in the capacity of blood cell integrins to adhere to fibrin in vivo and in vitro is striking and implies the existence of a physiologic antiadhesive mechanism. The surfaces of fibrin clots in the circulation are continually exposed to plasma proteins, several of which can bind fibrin and influence cell adhesion. Recently, we have demonstrated that adsorption of soluble fibrinogen on the surface of a fibrin clot results in its deposition as a soft multilayer matrix, which prevents attachment of blood cells. In the present study, we demonstrate that another plasma protein, plasminogen, which is known to accumulate in the superficial layer of fibrin, exerts an antiadhesive effect. After being coated with plasminogen, the surfaces of fibrin clots became essentially non-adhesive for U937 monocytic cells, blood monocytes, and platelets. The data revealed that activation of fibrin-bound plasminogen by the plasminogen-activating system assembled on adherent cells resulted in the generation of plasmin, which decomposed the superficial fibrin layer, resulting in cell detachment under flow. The surfaces generated after the initial cell adhesion remained non-adhesive for subsequent attachment of leukocytes and platelets. We propose that the limited degradation of fibrin by plasmin generated by adherent cells loosens the fibers on the clot surface, producing a mechanically unstable substrate that is unable to support firm integrin-mediated cell adhesion.


Effect of sulfur dioxide inhalation on erythrocyte antioxidant status and lipid peroxidation in experimental diabetes.
The effect of sulfur dioxide (SO(2) ) on red cell antioxidant status and lipid peroxidation was examined in this research. Forty healthy male albino rats, aged three months, were divided into four equal groups: Control (C), SO(2) +C (CSO(2) ), diabetic (D) and SO(2) +D (DSO(2) ). Experimental diabetes mellitus was induced by i.v injection of alloxan with a dose of 50 mg/kg body weight. Ten ppm SO(2) was administered to the animals of SO(2) exposed groups in an exposure chamber for one hr/day x 7 days/wk x 6wks while other groups were exposed to filtered air in the same condition. SO(2) exposure, while markedly decreasing Cu, Zn-Superoxide dismutase (Cu, Zn-SOD) activity, significantly increased glutathione peroxidase (GSH-Px), catalase (CAT), glutathione (GSH) and glutathione-s-transferase (GST) activities and TBARS values in CSO(2) and DSO(2) groups compared with their respective control groups. From these results, it could be concluded that adaptative changes occurred in antioxidant systems that may counteract the free radical effect of SO(2) in the experimental groups.


[Problems and prospects of space pharmacology].
The paper reviews the Soviet and foreign literature on the pharmacological support of manned space flights with emphasis on pharmacological prophylaxis and therapy of adverse effects of the space environment. The paper discusses future development of pharmacological studies as part of space medicine.


[Renal osteodystrophy Guidelines].
Renal ostedystrophy (ROD) is a major long-term complication in uremic patients. Bone histomorphometry still remains the gold standard for the diagnosis of ROD. However, the low acceptance grade by patients makes bone biopsy a rarely performed and not easily repeatable investigation. No other instrumental assessment has been proved as yet to have sufficient sensitivity for ROD diagnosis. Many biochemical markers have been proposed for a diagnostic role, but few have a real predictive diagnostic value. Serum intact PTH (i-PTH) levels are thought to represent a good predictor of bone lesions. However, although a i-PTH level greater than 450 pg/mL and lower than 120 pg/mL may well predict high and low bone turnover disease respectively, in the wide range of values defined by the above border levels i-PTH does not have a predictive role for ROD. There is as yet no definite proof that the recently developed PTH assays might increase their diagnostic sensitivity. Bone alkaline phosphatase is a more reliable index of bone turnover than i-PTH levels. With regards to Al overload, given that an iron overburden is excluded, serum Al levels lower than 30 ug/L are seldom associated with increased Al deposition; conversely, levels above 60 mg/L are highly diagnostic for Al overload. In the latter condition, a DFO test is recommended. The main goals of ROD treatment are a) to maintain serum i-PTH levels between 120 and 150 pg/mL; b) to bring the phosphate (Pi) concentration under 5.5 mg/dL, Ca concentration between 9.2 and 10.4 mg/dL, and the Ca x Pi product under 55 mg/dL; c) to bring Al concentration under 20 ug/L; and d) to target serum bicarbonate levels between 20 and 24 mmol/L. The main therapeutic approaches include: Dietary Pi intake control (< 1200 mg/day). Intestinal phosphate binding using calcium salts and sevelamer. Calcium salts must be used at a dosage that avoids Ca overload (< 23 g/day). If Pi control is not reached, Mg and Al salts may be added at a dose lower than 2 g/day and for less than 3 months. Appropriate dialysis dose (KT/V > 1.2) and dialysis time (consider increased dialysis duration or session number for a week). Ca concentration in dialysate and infusion fluids can range between 1.25 and 2.00 mmol/L, according to the dialysis technique and to maintain an appropriate Ca balance. Vitamin D should not be used when i-PTH levels are < 120 pg/mL and/or serum Ca > 11 mg/dL and/or Pi > 6.5 mg/dL. The vitamin D dose should be proportional to PTH levels, with a larger dose given as a bolus (23 ug twice or three times per week). The intravenous route should be preferred in the case of very high i-PTH (> 700 pg/mL) or after 3 months of unsuccessful oral treatment. PTX should be prescribed on the basis of the clinical, biochemical, and instrumental data. A 7/8 PTX, when possible, is the most advisable procedure. Bone transplant disease (BTD) is caused by a combination of previous uremic ROD and bone lesions occurring after renal transplantation, mainly secondary to steroid effect. The main clinical result of BTD is the osteopenicosteoporotic syndrome, which frequently results in bone fractures. The most effective treatment of BTD is a reduction of the cumulative steroid dose. No strong evidence has been produced as yet for a preventive role of either bisphosphonates or vitamin D supplementation on BTD.


Increased glucose uptake precedes hyperinsulinemia in awake endotoxemic minipigs.
The role of hyperinsulinemia in the development of hypoglycemia was evaluated in awake Yucatan minipigs. Eight adult minipigs were fitted with jugular, portal, and hepatic vein and carotid artery catheters, and hepatic artery and portal vein flow cuffs for determination of transhepatic kinetics and insulin secretion. Three days later they were infused with E. coli endotoxin at 15 micrograms/kg/hr. Transient hyperinsulinemia was preceded by an elevation of the rate of glucose uptake (Rd) determined from [6-3H] glucose-specific activity. This finding suggested that hyperinsulinemia might be caused by, and then be contributory to, increased Rd. If an increase in glucose uptake was also experienced by pancreatic beta-cells, the existing glycemic state could be overestimated and an inappropriate insulin release elicited.


Cytologic diagnosis of cholesterol granuloma. A case report.
Cholesterol granuloma is a rare lesion of the orbit occurring in adults and must be distinguished from epidermoid cholesteatoma and other mass lesions of the orbit. We report the case of a 31-year-old man who presented to our institution with right eye proptosis secondary to a soft tissue mass. At surgery, cyst fluid from the lesion was sent for cytologic evaluation. The fluid was remarkable for numerous inflammatory cells, blood breakdown products, multinucleate giant cells and cholesterol crystals. The absence of squamous- or respiratory-type epithelium led to a diagnosis of cholesterol granuloma. This was confirmed by histologic examination of the tissue submitted. The differential diagnosis and cytologic features of orbital lesions are discussed.


Targeted genotyping by sequencing: a new way to genome profile the cat.
Targeted GBS is a recent approach for obtaining an effective characterization for hundreds to thousands of markers. The high throughput of next-generation sequencing technologies, moreover, allows sample multiplexing. The aims of this study were to (i) define a panel of single nucleotide polymorphisms (SNPs) in the cat, (ii) use GBS for profiling 16 cats, and (iii) evaluate the performance with respect to the inference using standard approaches at different coverage thresholds, thereby providing useful information for designing similar experiments. Probes for sequencing 230 variants were designed based on the Felis_catus_8.0. 8.0 genome. The regions comprised anonymous and non-anonymous SNPs. Sixteen cat samples were analysed, some of which had already been genotyped in a large group of loci and one having been whole-genome sequenced in the 99_Lives Cat Genome Sequencing Project. The accuracy of the method was assessed by comparing the GBS results with the genotypes already available. Overall, GBS achieved good performance, with 92-96% correct assignments, depending on the coverage threshold used to define the set of trustable genotypes. Analyses confirmed that (i) the reliability of the inference of each genotype depends on the coverage at that locus and (ii) the fraction of target loci whose genotype can be inferred correctly is a function of the total coverage. GBS proves to be a valid alternative to other methods. Data suggested a depth of less than 11× is required for greater than 95% accuracy. However, sequencing depth must be adapted to the total size of the targets to ensure proper genotype inference.


Glyphosate commercial formulation causes cytotoxicity, oxidative effects, and apoptosis on human cells: differences with its active ingredient.
In the present study, the effects on oxidative balance and cellular end points of glyphosate, aminomethylphosphonic acid (AMPA), and a glyphosate formulation (G formulation) were examined in HepG2 cell line, at dilution levels far below agricultural recommendations. Our results show that G formulation had toxic effects while no effects were found with acid glyphosate and AMPA treatments. Glyphosate formulation exposure produced an increase in reactive oxygen species, nitrotyrosine formation, superoxide dismutase activity, and glutathione (GSH) levels, while no effects were observed for catalase and GSH-S-transferase activities. Also, G formulation triggered caspase 3/7 activation and hence induced apoptosis pathway in this cell line. Aminomethylphosphonic acid exposure produced an increase in GSH levels while no differences were observed in other antioxidant parameters. No effects were observed when the cells were exposed to acid glyphosate. These results confirm that G formulations have adjuvants working together with the active ingredient and causing toxic effects that are not seen with acid glyphosate.


Medical disorders in Alzheimer's disease and vascular dementia.
The clinical and postmortem findings of 29 patients with Alzheimer's disease were evaluated and compared to the findings of 19 patients with vascular dementia. The patients with Alzheimer's disease had received treatment for an average of 2.0 internal medical disorders, the patients with vascular dementia for 2.1 disorders. The average number of medical diseases found at postmortem was 3.7 in the group with Alzheimer's dementia and 4.1 in vascular dementia. Apart from a marginally increased rate of peripheral occlusive artery disease in vascular dementia, the proportions of the most common disorders of the cardiovascular, genitourinary, gastrointestinal and endocrinological disorders and the most common causes of death did not differ significantly between these groups. Therefore patients with Alzheimer's disease should receive the same internal medical attention as patients with vascular dementia.


Kinetic complexity of chloroplastal deoxyribonucleic acid and mitochondrial deoxyribonucleic acid from higher plants.
1. Chloroplasts and mitochondria were isolated by aqueous and non-aqueous cell-fractionation techniques. In a variety of higher plants the mitochondrial DNA bands in a caesium chloride gradient at 1.706g.cm.(-3), whereas chloroplastal DNA has a buoyant density of 1.697g.cm.(-3). 2. In total cellular DNA of moderate molecular weight, the chloroplastal DNA is found within the Gaussian distribution of the nuclear DNA and is not resolved as a satellite. 3. Both chloroplastal DNA and mitochondrial DNA from lettuce renature rapidly. 4. The kinetic complexity of mitochondrial DNA is > 10(8) daltons. 5. Chloroplastal DNA is made up from fast and slow renaturing sequences with kinetic complexities of 3x10(6) and 1.2x10(8) daltons respectively. 6. From the discrepancy between analytical and kinetic complexity it is concluded that chloroplastal DNA is extensively reiterated.


Transformation and transfection in lysogenic strains of Bacillus subtilis 168.
Strains of Bacillus subtilis lysogenic for either temperate bacteriophage phi105 or SPO2 were reduced to less than 1.0% of the level of transformation of the nonlysogenic strains. Strains lysogenic for both phi105 and SPO2 are virtually nontransformable, indicating that the effect of lysogeny is additive. Lysogenic cultures transfected at essentially wild-type levels with deoxyribonucleic acid (DNA) isolated from bacteriophages phi29 and SPO1. The residual transformation and transfection achieved by the lysogenic cultures changed dramatically during growth in SPII medium, whereas nonlysogenic strains remained competent for 5 hr in SPII medium. Despite a marked reduction in transformation, lysogenic cultures initially irreversibly bound as much DNA as nonlysogenic cultures. After 60 min in SPII medium, there was a rapid decrease in the capacity of lysogenic cells to bind DNA irreversibly. These results, as discussed, indicate that the inhibition of transformation is probably due to an alteration of the cell surface or a differential inactivation of bacterial genes after lysogenic conversion.


Bulge migration of the malondialdehyde OPdG DNA adduct when placed opposite a two-base deletion in the (CpG)3 frameshift hotspot of the Salmonella typhimurium hisD3052 gene.
The OPdG adduct N (2)-(3-oxo-1-propenyl)dG, formed in DNA exposed to malondialdehyde, was introduced into 5'-d(ATCGC XCGGCATG)-3'.5'-d(CATGCCGCGAT)-3' at pH 7 (X = OPdG). The OPdG adduct is the base-catalyzed rearrangement product of the M 1dG adduct, 3-(beta- d-ribofuranosyl)pyrimido[1,2- a]purin-10(3 H)-one. This duplex, named the OPdG-2BD oligodeoxynucleotide, was derived from a frameshift hotspot of the Salmonella typhimuium hisD3052 gene and contained a two-base deletion in the complementary strand. NMR spectroscopy revealed that the OPdG-2BD oligodeoxynucleotide underwent rapid bulge migration. This hindered its conversion to the M 1dG-2BD duplex, in which the bulge was localized and consisted of the M 1dG adduct and the 3'-neighbor dC [ Schnetz-Boutaud, N. C. , Saleh, S. , Marnett, L. J. , and Stone, M. P. ( 2001) Biochemistry 40, 15638- 15649 ]. The spectroscopic data suggested that bulge migration transiently positioned OPdG opposite dC in the complementary strand, hindering formation of the M 1dG-2BD duplex, or alternatively, reverting rapidly formed intermediates in the OPdG to M 1dG reaction pathway when dC was placed opposite from OPdG. The approach of initially formed M 1dG-2BD or OPdG-2BD duplexes to an equilibrium mixture of the M 1dG-2BD and OPdG-2BD duplexes was monitored as a function of time, using NMR spectroscopy. Both samples attained equilibrium in approximately 140 days at pH 7 and 25 degrees C.


Comparative multilocus phylogeography of two Palaearctic spruce bark beetles: influence of contrasting ecological strategies on genetic variation.
While phylogeographic patterns of organisms are often interpreted through past environmental disturbances, mediated by climate changes, and geographic barriers, they may also be strongly influenced by species-specific traits. To investigate the impact of such traits, we focused on two Eurasian spruce bark beetles that share a similar geographic distribution, but differ in their ecology and reproduction. Ips typographus is an aggressive tree-killing species characterized by strong dispersal, whereas Dendroctonus micans is a discrete inbreeding species (sib mating is the rule), parasite of living trees and a poor disperser. We compared genetic variation between the two species over both beetles' entire range in Eurasia with five independent gene fragments, to evaluate whether their intrinsic differences could have an influence over their phylogeographic patterns. We highlighted widely divergent patterns of genetic variation for the two species and argue that the difference is indeed largely compatible with their contrasting dispersal strategies and modes of reproduction. In addition, genetic structure in I. typographus divides European populations in a northern and a southern group, as was previously observed for its host plant, and suggests past allopatric divergence. A long divergence time was estimated between East Asian and other populations of both species, indicating their long-standing presence in Eurasia, prior to the last glacial maximum. Finally, the strong population structure observed in D. micans for the mitochondrial locus provides insights into the recent colonization history of this species, from its native European range to regions where it was recently introduced.


Amplification of the RARA gene in acute myeloid leukemia: significant finding or coincidental observation?
Oncogene amplification resulting in aberrant expression, although common in solid tumors, is rare in acute myeloid leukemia (AML) and is mostly associated with amplification of MYC, RUNX1, and MLL genes. Retinoic acid receptor alpha (RARA) and other target sequences at 17p11.2 often represent the amplicons expressed in breast cancer, not in AML. We present a unique case of a 59-year-old female with a history of breast cancer, now presenting with pancytopenia and bilateral infiltration with effusion in nodules of the right upper lobe of the lung. She was diagnosed with AML-M5. Chromosome analysis demonstrated a hypodiploid clone with complex numerical/structural abnormalities including 5q deletion, monosomy 7, as well as structurally rearranged chromosome 11 and several marker chromosomes. Fluorescence in situ hybridization (FISH) analysis showed amplification of RARA, loss of 7q, monosomy 7, loss of DEK (6p23), and additional copies of NUP214 (9q34) and MLL (11q23). Additional FISH studies showed both ERBB2 and TOP2A genes, which were co-amplified on one of the marker chromosomes. The follow-up bone marrow did not yield any metaphases, but FISH was normal for all probes, including RARA. After a short remission, the patient relapsed and showed clonal evolution. Additional case reports are necessary to assess whether RARA amplification in hematologic malignancies serves as an independent prognostic factor.


Responsiveness of multiple patient-reported outcome measures for acute postsurgical pain: primary results from the international multi-centre PROMPT NIT-1 study.
Postsurgical outcome measures are crucial to define the efficacy of perioperative pain management; however, it is unclear which are most appropriate. We conducted a prospective study aiming to assess sensitivity-to-change of patient-reported outcome measures assessing the core outcome set of domains pain intensity (at rest/during activity), physical function, adverse events, and self-efficacy. Patient-reported outcome measures were assessed preoperatively, on day 1 (d1), d3, and d7 after four surgical procedures (total knee replacement, breast surgery, endometriosis-related surgery, and sternotomy). Primary outcomes were sensitivity-to-change of patient-reported outcome measures analysed by correlating their changes (d1-d3) with patients' global impression of change and patients' specific impression of change items as anchor criteria. Secondary outcomes included identification of baseline and patient characteristic variables explaining variance in change for each of the scales and descriptive analysis of various patient-reported outcome measures from different domains and after different surgeries. Of 3322 patients included (18 hospitals, 10 countries), data from 2661 patients were analysed. All patient-reported outcome measures improved on average over time; the median calculated sensitivity-to-change for all patient-reported outcome measures (overall surgeries) was 0.22 (range: 0.07-0.31, scale: 0-10); all changes were independent of baseline data or patient characteristics and similar between different procedures. Pain-related patient-reported outcome measures have low to moderate sensitivity-to-change; those showing higher sensitivity-to-change from the same domain should be considered for inclusion in a core outcome set of patient-reported outcome measures to assess the effectiveness and efficacy of perioperative pain management.