Gold excretion and retention during auranofin treatment: a preliminary report.
Auranofin, an oral gold compound, was administered to 12 patients with rheumatoid arthritis using 2 dosage schedules (3 mg or 1 mg twice daily for 8 weeks, and then once daily for 18 weeks). In addition to outpatient clinic monitoring, all patients were admitted to a metabolic ward for 3 days for collection of 24 hr daily urine and feces. Gold content of excreta was determined by atomic absorption spectroscopy. Seventy-three per cent of the administered gold was recovered in the urine and feces of patients receiving 3 mg b.i.d., and all the gold was recovered in those taking 1 mg b.i.d. Ninety-five per cent of the recovered gold was in the feces and 5% was in the urine. These findings contrast with those observed during intramuscular (gold sodium thiomalate) chrysotherapy: 40% of the injected dose was recovered, 70% in urine, 30% in feces. Less tissue gold retention occurred with oral gold than with parenteral therapy. Following 20 weeks of auranofin (6 mg/day) chrysotherapy approximately 66 mg of gold was retained. By comparison, 300 mg of gold was retained after injectable gold sodium thiomalate treatment. The significance of these findings is discussed.


Influence of fine structure of lipid A on Limulus amebocyte lysate clotting and toxic activities.
We examined the relationship between the fine structure of lipid A and the toxicity of endotoxin or lipopolysaccharides as measured by the Limulus amebocyte lysate (LAL), rabbit pyrogenicity, chicken embryo lethal dose, and dermal Shwartzman reaction tests. Lipid A and lipid A-like compounds obtained from deep-rough mutants of Salmonella spp. and Escherichia coli had a wide range of structural variations. These compounds included native lipopolysaccharides, diphosphoryl and monophosphoryl lipid A's, and lipid X (a monosaccharide). The LAL test was positive for all lipids tested with lysates from Travenol Laboratories and from Associates of Cape Cod (2.9 X 10(3) to 2.6 X 10(7) endotoxin units per mg), except for O-deacylated and dephosphorylated lipid X, which were negative. The Mallinckrodt lysate gave negative tests for lipid X. In the rabbit pyrogenicity and chicken embryo lethal dose tests, only native lipopolysaccharide and diphosphoryl lipid A's were judged toxic. The Shwartzman reaction was positive for a specific purified diphosphoryl lipid A (thin-layer chromatography-3 fraction) but negative for the purified monophosphoryl lipid A (also a thin-layer chromatography-3 fraction). These results show that the LAL test is not a valid measure of all parameters of toxicity of a lipid A or lipid A-like compound and can yield false-positive results. However, these findings are not in conflict with the widespread use of the LAL assay for pyrogens in the pharmaceutical industry since a good correlation exists between LAL results and pyrogenicity when undegraded endotoxin is evaluated in parallel assays.


CXCR4-expressing Mist1+ progenitors in the gastric antrum contribute to gastric cancer development.
Mist1 was recently shown to identify a discrete population of stem cells within the isthmus of the oxyntic gland within the gastric corpus. Chief cells at the base of the gastric corpus also express Mist1. The relevance of Mist1 expression as a marker of specific cell populations within the antral glands of the distal stomach, however, is unknown. Using Mist1-CreERT mice, we revealed that Mist1+ antral cells, distinct from the Mist1+ population in the corpus, comprise long-lived progenitors that reside within the antral isthmus above Lgr5+ or CCK2R+ cells. Mist1+ antral progenitors can serve as an origin of antral tumors induced by loss of Apc or MNU treatment. Mist1+ antral progenitors, as well as other antral stem/progenitor population, express Cxcr4, and are located in close proximity to Cxcl12 (the Cxcr4 ligand)-expressing endothelium. During antral carcinogenesis, there is an expansion of Cxcr4+ epithelial cells as well as the Cxcl12+ perivascular niche. Deletion of Cxcl12 in endothelial cells or pharmacological blockade of Cxcr4 inhibits antral tumor growth. Cxcl12/Cxcr4 signaling may be a potential therapeutic target.


Supraumbilical pyloromyotomy: a unique indication for antimicrobial prophylaxis.
The umbilical fold incision for infantile hypertrophic pyloric stenosis provides a convenient exposure and cosmetically appealing scar. This study investigates the possible difference in infection rates between traditional and supraumbilical approaches for pyloromyotomy. All patients who underwent pyloromyotomy for infantile hypertrophic pyloric stenosis at a tertiary pediatric hospital were reviewed. Baseline wound infection rate was determined through review of patients with right upper quadrant incisions (group 1). A nonrandomized comparison was performed between patients with a supraumbilical approach (group 2) and those undergoing supraumbilical incisions after prophylactic antibiotic administration (group 3). Complete records were reviewed on 384 patients over a 6-year period. Demographics and preoperative factors were similar among groups. The rate of infection in group 1 (n = 258) was 2.3%. With introduction of the supraumbilical approach, there was a statistically significant increase in wound infection rate to 7.0% (chi 2 ; group 1 vs group 2, P < .05). The use of prophylactic antibiotics with a supraumbilical approach reduced this rate of infection back to 2.3% (chi 2 ; group 1 vs group 3, P < 1.0 and group 2 [n = 85] vs group 3 [n = 42], P < .3). The risk of wound infection by classic pyloromyotomy of 2.3% is significantly increased with an open supraumbilical approach. The use of prophylactic antibiotics reduces this risk of wound infection.


Powerful, onset inhibition in the ventral nucleus of the lateral lemniscus.
The function of the ventral nucleus of the lateral lemniscus (VNLL), a secondary processing site within the auditory brain stem, is unclear. It is known to be a major source of inhibition to the inferior colliculus (IC). It is also thought to play a role in coding the temporal aspects of sound, such as onsets and the periodic components of complex stimuli. In vivo intracellular recordings from VNLL neurons (n = 56) in urethane anesthetized rats revealed the presence of large-amplitude, short-duration, onset inhibition in a subset of neurons (14.3%). This inhibition occurred before the first action potential (AP) elicited by noise or tone bursts, was broadly tuned to tonal frequency and was shown to delay the first AP. Our data suggest it is a result of an intrinsic circuit activated by the octopus cell pathway originating in the contralateral cochlear nucleus; this pathway is known to convey exquisitely timed and broadly tuned onset information. This powerful inhibition within the VNLL appears to control the timing of this structure's inhibitory output to higher centers, which has important auditory processing outcomes. The circuit also provides a pathway for fast, broadly tuned, onset inhibition to the IC.


Suppression of anger and gastric emptying in patients with functional dyspepsia.
Psychologic distress and gastric motor dysfunction have both been implicated in the pathogenesis of functional (non-ulcer) dyspepsia (FD). This study assesses the association between psychologic factors and gastric emptying in 28 FD patients. Subjects completed an extensive range of psychologic questionnaires and underwent dual-isotope scintigraphic assessment of solid and liquid gastric emptying. Attempts to resist, control, suppress, and hold in anger, to adopt a fighting spirit whilst dealing with chronic stressors, and manifest unhappiness were predictors of prolonged gastric emptying. These findings suggest that psychologic factors may be important in the aetiology of gastric stasis and subsequent upper gastrointestinal symptoms in patients with functional dyspepsia.


Geochemically induced shifts in catabolic energy yields explain past ecological changes of diffuse vents in the East Pacific Rise 9°50'N area.
The East Pacific Rise (EPR) at 9°50'N hosts a hydrothermal vent field (Bio9) where the change in fluid chemistry is believed to have caused the demise of a tubeworm colony. We test this hypothesis and expand on it by providing a thermodynamic perspective in calculating free energies for a range of catabolic reactions from published compositional data. The energy calculations show that there was excess H2S in the fluids and that oxygen was the limiting reactant from 1991 to 1997. Energy levels are generally high, although they declined in that time span. In 1997, sulfide availability decreased substantially and H2S was the limiting reactant. Energy availability dropped by a factor of 10 to 20 from what it had been between 1991 and 1995. The perishing of the tubeworm colonies began in 1995 and coincided with the timing of energy decrease for sulfide oxidizers. In the same time interval, energy availability for iron oxidizers increased by a factor of 6 to 8, and, in 1997, there was 25 times more energy per transferred electron in iron oxidation than in sulfide oxidation. This change coincides with a massive spread of red staining (putative colonization by Fe-oxidizing bacteria) between 1995 and 1997.For a different cluster of vents from the EPR 9°50'N area (Tube Worm Pillar), thermodynamic modeling is used to examine changes in subseafloor catabolic metabolism between 1992 and 2000. These reactions are deduced from deviations in diffuse fluid compositions from conservative behavior of redox-sensitive species. We show that hydrogen is significantly reduced relative to values expected from conservative mixing. While H2 concentrations of the hydrothermal endmember fluids were constant between 1992 and 1995, the affinities for hydrogenotrophic reactions in the diffuse fluids decreased by a factor of 15 and then remained constant between 1995 and 2000. Previously, these fluids have been shown to support subseafloor methanogenesis. Our calculation results corroborate these findings and indicate that the 1992-1995 period was one of active growth of hydrogenotrophic communities, while the system was more or less at steady state between 1995 and 2000.


Gut-liver interaction in glutamine homeostasis: portal ammonia role in uptake and metabolism.
The role of ammonia released by the gut on hepatic glutamine handling and metabolism was studied in postabsorptive anesthesized male Sprague-Dawley rats at spontaneous and elevated arterial glutamine concentrations. Glutamine handling and metabolite release across both organ beds were studied using arteriovenous concentration differences and simultaneously measured portal and hepatic venous plasma flows. At the spontaneous arterial glutamine load, fractional glutamine extraction, FE-Gln, by the gut and the liver was 24 and 10%, respectively. At the elevated glutamine load, gut glutamine uptake doubled, while FE-Gln remained at 24%; however, portal ammonia and alanine increased and decreased, respectively. In response, hepatic FE-Gln increased to 28% with a large release of glutamate and urea. The role of portal ammonia in modulating hepatic glutamine uptake was studied by infusing NH4HCO3 directly into the portal vein. Increasing the portal load promptly stimulated hepatic glutamine uptake and glutamate and urea release. Mitochondria isolated from these livers produced more glutamate from glutamine, suggesting ammonia activation of hepatic glutaminase flux; in addition, citrulline formation increased, suggesting a coupling of glutaminase flux to urea synthesis. The results are consistent with portal ammonia release acting as a key informational molecule in interorgan glutamine flow.


Novel targets for antibiotics--an update.
The emergence of widespread antibiotic resistance as an impediment in the treatment of bacterial diseases is of growing concern. In some instances, clinicians are left with few or no antibiotics for treatment of infections and this problem will more than likely grow in magnitude. One approach to get around the problem of antibiotic resistance is to develop new drugs with novel targets and mechanisms of action. Due to the 'newness' of these novel targets as therapeutic targets, the likelihood that resistance will initially be widespread is low. Three approaches are discussed in this overview: discovery of new essential genes that are expressed exclusively in vivo development of compounds that act on global bacterial gene regulators; and interference with virulence determinants. By exploiting virulence related attributes or genes expressed exclusively in vivo, the risk of resistance is reduced since inhibiting these products will probably alter the ecology (habitats) of these organisms rather than causing direct cell death. This might also lead to a selective targeting of pathogens with the beneficial consequence of ignoring organisms growing in their normal habitat, such as in the gastrointestinal tract or skin.


Factors controlling smooth-muscle cell proliferation.
Injury to the arterial wall normally elicits a rapid and significant increase in smooth-muscle cell (SMC) replication with the subsequent development of intimal lesions. A variety of factors have been proposed to control SMC replication, but recent work has highlighted the role of basic fibroblast growth factor (bFGF) and platelet-derived growth factor in this process. In the carotid artery of the uninjured rat, we have shown that the SMCs express mRNA for bFGF and that bFGF can be readily extracted from these arteries. Following mechanical injury to the artery, ie, after balloon injury, we suggested that bFGF is released from damaged cells and then stimulates adjacent SMCs. In support of this concept, the infusion of a blocking antibody to bFGF was found to significantly inhibit the early SMC replication induced by use of a balloon catheter. The addition of the antibody at the time of injury, however, did not inhibit the development of intimal lesions. In contrast, studies by us and other investigators have shown that platelet-derived growth factor is not directly important for SMC replication after balloon injury, but that it plays a key role in stimulating the migration of SMCs into the intima. Intimal SMC replication was not inhibited with antibodies to either bFGF or platelet-derived growth factor. Therefore, while significant inroads have been made in understanding the initial events, we still do not fully understand all the processes involved in the proliferation of arterial intimal lesions.


Relationship Between Mobile Digital Sensor Monitoring and Perioperative Outcomes: Systematic Review.
Monitoring surgical recovery has traditionally been confined to metrics measurable within the hospital and clinic setting. However, commercially available mobile sensors are now capable of extending measurements into a patient's home. As these sensors were developed for nonmedical applications, their clinical role has yet to be established. The aim of this systematic review is to evaluate the relationship between data generated by mobile sensors and postoperative outcomes. The objective of this study is to describe the current use of mobile sensors in the perioperative setting and the correlation between their data and clinical outcomes. A systematic search of EMBASE, MEDLINE, and Cochrane Library from inception until April 2019 was performed to identify studies of surgical patients monitored with mobile sensors. Sensors were considered if they collected patient metrics such as step count, temperature, or heart rate. Studies were included if patients underwent major surgery (≥1 inpatient postoperative day), patients were monitored using mobile sensors in the perioperative period, and the study reported postoperative outcomes (ie, complications and hospital readmission). For studies including step count, a pooled analysis of the step count per postoperative day was calculated for the complication and noncomplication cohorts using mean and a random-effects linear model. The Grading of Recommendations, Assessment, Development, and Evaluation tool was used to assess study quality. From 2209 abstracts, we identified 11 studies for review. Reviewed studies consisted of either prospective observational cohorts (n=10) or randomized controlled trials (n=1). Activity monitors were the most widely used sensors (n=10), with an additional study measuring temperature, respiratory rate, and heart rate (n=1). Low step count was associated with worse postoperative outcomes. A median step count of around 1000 steps per postoperative day was associated with adverse surgical outcomes. Within the studies, there was heterogeneity between the type of surgery and type of reported postoperative outcome. Despite significant heterogeneity in the type of surgery and sensors, low step count was associated with worse postoperative outcomes across surgical specialties. Further studies and standardization are needed to assess the role of mobile sensors in postoperative care, but a threshold of approximately 1000 steps per postoperative day warrants further investigation.


AAPM WGDCAB Report 372: A joint AAPM, ESTRO, ABG, and ABS report on commissioning of model-based dose calculation algorithms in brachytherapy.
The introduction of model-based dose calculation algorithms (MBDCAs) in brachytherapy provides an opportunity for a more accurate dose calculation and opens the possibility for novel, innovative treatment modalities. The joint AAPM, ESTRO, and ABG Task Group 186 (TG-186) report provided guidance to early adopters. However, the commissioning aspect of these algorithms was described only in general terms with no quantitative goals. This report, from the Working Group on Model-Based Dose Calculation Algorithms in Brachytherapy, introduced a field-tested approach to MBDCA commissioning. It is based on a set of well-characterized test cases for which reference Monte Carlo (MC) and vendor-specific MBDCA dose distributions are available in a Digital Imaging and Communications in Medicine-Radiotherapy (DICOM-RT) format to the clinical users. The key elements of the TG-186 commissioning workflow are now described in detail, and quantitative goals are provided. This approach leverages the well-known Brachytherapy Source Registry jointly managed by the AAPM and the Imaging and Radiation Oncology Core (IROC) Houston Quality Assurance Center (with associated links at ESTRO) to provide open access to test cases as well as step-by-step user guides. While the current report is limited to the two most widely commercially available MBDCAs and only for 192 Ir-based afterloading brachytherapy at this time, this report establishes a general framework that can easily be extended to other brachytherapy MBDCAs and brachytherapy sources. The AAPM, ESTRO, ABG, and ABS recommend that clinical medical physicists implement the workflow presented in this report to validate both the basic and the advanced dose calculation features of their commercial MBDCAs. Recommendations are also given to vendors to integrate advanced analysis tools into their brachytherapy treatment planning system to facilitate extensive dose comparisons. The use of the test cases for research and educational purposes is further encouraged.


Fatty Acids and Eicosanoids Change during High-Fiber Diet in NAFLD Patients-Randomized Control Trials (RCT).
Non-alcoholic fatty liver disease (NAFLD) is a wide spectrum condition characterized by excessive liver fat accumulation in people who do not abuse alcohol. There is no effective medical treatment for NAFLD; therefore, most important recommendations to reduce liver steatosis are diet and lifestyle, including proper physical activity. The aim of our study was to analyze the fatty acids and eicosanoids changes in the serum of patients who consumed high-fiber rolls for 8 weeks. The group of 28 Caucasian participants was randomly divided into two groups, those who received 24 g of fiber/day-from 2 buns of 12 g each (n = 14), and those who received 12 g of fiber/day-from 2 buns of 6 g (n = 14). At the beginning and on the last visit of the 8-week intervention, all patients underwent NAFLD evaluation, biochemical parameter measurements, and fatty acids and eicosanoids evaluation. Patients who received 12 g of fiber had significantly reduced liver steatosis and body mass index. In the group who received 24 g of fiber/day, we observed a trend to liver steatosis reduction (p = 0.07) and significant decrease in aspartate aminotransferase (p = 0.03) and total cholesterol (p = 0.03). All changes in fatty acid and eicosanoids profile were similar. Fatty acids analysis revealed that extra fiber intake was associated with a significant increase in monounsaturated fatty acids and decrease in saturated fatty acids. Moreover, both groups showed increased concentration of gamma linoleic acid and docosahexaenoic acid. We also observed reduction in prostaglandin E2. Our study revealed that a high amount of fiber in the diet is associated with a reduction in fatty liver, although this effect was more pronounced in patients in the lower fiber group. However, regardless of the amount of fiber consumed, we observed significant changes in the profile of FAs, which may reflect the positive changes in the lipids liver metabolism. Regardless of the amount of fiber consumed, patients decreased the amount of PGE2, which may indicate the lack of disease progression associated with the development of inflammation.


Socioeconomic modifiers of genetic and environmental influences on body mass index in adult twins.
Individual measures of socioeconomic status (SES) suppress genetic variance in body mass index (BMI). Our objective was to examine the influence of both individual-level (i.e., educational attainment, household income) and macrolevel (i.e., neighborhood socioeconomic advantage) SES indicators on genetic contributions to BMI. The study used education level data from 4,162 monozygotic (MZ) and 1,900 dizygotic (DZ) same-sex twin pairs (64% female), income level data from 3,498 MZ and 1,534 DZ pairs (65% female), and neighborhood-level socioeconomic deprivation data from 2,327 MZ and 948 DZ pairs (65% female). Covariates included age (M = 40.4 ± 17.5 years), sex, and ethnicity. The cotwin control model was used to evaluate the mechanisms through which SES influences BMI (e.g., through genetic vs. environmental pathways), and a gene-by-environment interaction model was used to test whether residual variance in BMI, after controlling for the main effects of SES, was moderated by socioeconomic measures. SES significantly predicted BMI. The association was noncausal, however, and instead was driven primarily through a common underlying genetic background that tended to grow less influential as SES increased. After controlling for the main effect of SES, both genetic and nonshared environmental variance decreased with increasing SES. The impact of individual and macrolevel SES on BMI extends beyond its main effects. The influence of genes on BMI is moderated by individual and macrolevel measures of SES, such that when SES is higher, genetic factors become less influential.


Chloride channels in myocytes from rabbit colon are regulated by a pertussis toxin-sensitive G protein.
Because of the high intracellular Cl- concentration ([Cl-]i) in gastrointestinal smooth muscle, receptor-mediated opening of Cl- channels at the cell resting potential could represent a plausible mechanism for initial receptor-mediated cell depolarization. To test this hypothesis, we characterized activation of large-conductance Cl- channels by the neurokinin-1 (NK-1) receptor agonist [Sar9,Met(O2)11]-substance P, by specific second messengers, and by direct G protein activation in myocytes isolated from the rabbit colon longitudinal muscle layer. In excised inside-out patches, large-conductance ion channels selective for Cl- over Na+ could be induced by holding the patch at pipette potentials values > 60 mV. The channel showed multiple smaller conductance states (< or = 20) but could open and close via a main gate. When the channel was fully open, its slope conductance was 300 pS, with substates as small as 15 pS, comparable to the predominant conductance observed in cell-attached patches. The voltage-activation profile for full conductance was bell-shaped with maximal open probability (Po) for channel opening of approximately 0 mV. In cell-attached patches, addition of the NK-1 agonist to pipette solution activated a channel that corresponded to a subconductance state of the maxi Cl- channel. The voltage-activation profile for this subconductance state showed a maximal Po value for membrane potentials of approximately 0 mV, with rapid inactivation at more positive and partial inactivation at more negative membrane potentials. In excised inside-out patches, both the full and smaller conductance states of the Cl- channel were activated by the nonhydrolyzable guanosine triphosphate analogue guanosine 5'-O-(3-thiotriphosphate) and inhibited by pertussis toxin (PTX), whereas [Ca2+]i increased channel activity only in concentrations > 1 mM. In cell-attached patches, addition of different Ca2+ ionophores resulted in channel activation in 10% of cells, and activators of protein kinase A or protein kinase C had no effect. These findings are consistent with the hypothesis of a possible role of G protein-coupled Cl- channels in receptor-mediated initial cell depolarization in longitudinal colonic smooth muscle.


Gray matter abnormalities in patients with premenstrual dysphoric disorder: an optimized voxel-based morphometry.
Although brain neurochemistry is thought to play a role in the development of premenstrual dysphoric disorder (PMDD), neuroimaging studies of PMDD are sparse. We examined the extent to which gray matter (GM) abnormalities were present in women with PMDD compared to healthy controls. 3.0T magnetic resonance imaging scans of 15 women with PMDD and 15 healthy controls were compared using optimized voxel-based morphometry (VBM) analysis. A regression analysis was used to assess the relationship between GM density and PMDD-symptom severity. Our results showed significantly increased GM density in the hippocampal cortex and significantly decreased GM density in the parahippocampal cortex among women with PMDD compared to healthy controls. However, these GM abnormalities were not significantly associated with the severity of PMDD. Our inferences of the relationships between structural alterations and PMDD are drawn from a small sample, which may have increased the likelihood of type I error. GM abnormalities in limbic and paralimbic cortices were found to be associated with the pathophysiology of PMDD. Etiology of PMDD is likely related to emotional processing and self-regulation. Our findings provide a basis of neurobiological model for PMDD.


[An evaluation of the nutritional status of patients with chronic kidney failure on hemodialysis via rapid-turnover proteins].
The evaluation of the nutritional state of patients on maintenance haemodialysis is one of the main aspects involved in the prescription of treatment, since malnutrition is frequent among these patients and is a very important risk factor. We studied the albumin levels and the levels of several rapid interchange proteins (prealbumin, transferrin, cholinesterase) in 106 patients with chronic renal failure on haemodialysis. The proteic catabolism rate (pcr) and total dose on normalized dialysis (KT/V) was also determined in these patients, in accordance with the kinetic urea model. Anthropometrical measurements were taken (dry weight following haemodialysis, skin fold of the triceps and muscular circumference of the arm) in 65 patients. The average levels of the proteins studied were within normal laboratory limits, except for albumin, which was slightly lower. The greater frequency of infranormal levels corresponded to albumin (57%); the protein least altered was prealbumin (14.7%), although 70.4% of patients showed lower levels of this protein compared to those considered as indicating a poor prognosis (30 mg/day). The estimated daily proteic intake, according to the proteic catabolism rate, was lower than the recommended rate in 58% of our patients, this was not correlated with any of the proteins studied, and was significantly lower in the group of patients whose dialysis dose was too low. Although the anthropomorphic parameters did not correlate with any protein, the average levels of prealbumin were significantly lower in patients with infranormal levels of dry weight and skin fold of the triceps. The albumin, prealbumin, transferrin and cholineserase levels were not affected by treatment with erithropoyetin, haemodialysis buffer bath or type of membrane used.


Identification of a claudin-4 and E-cadherin score to predict prognosis in breast cancer.
The elevated expression of claudins (CLDN) and E-cadherin (CDH-1) was found to correlate with poor prognostic features. Our aim was to perform a comprehensive analysis to assess their potential to predict prognosis in breast cancer. The expression of CLDN-1, -3-5, -7, -8, -10, -15, -18, and E-cadherin at the mRNA level was evaluated in correlation with survival in datasets containing expression measurements of 1809 breast cancer patients. The breast cancer tissues of 197 patients were evaluated with tissue microarray technique and immunohistochemical method for CLDN-1-5, -7, and E-cadherin protein expression. An additional validation set of 387 patients was used to test the accuracy of the resulting prognostic score. Based on the bioinformatic screening of publicly-available datasets, the metagene of CLDN-3, -4, -7, and E-cadherin was shown to have the most powerful predictive power in the survival analyses. An immunohistochemical protein profile consisting of CLDN-2, -4, and E-cadherin was able to predict outcome in the most effective manner in the training set. Combining the overlapping members of the above two methods resulted in the claudin-4 and E-cadherin score (CURIO), which was able to accurately predict relapse-free survival in the validation cohort (P = 0.029). The multivariate analysis, including clinicopathological variables and the CURIO, showed that the latter kept its predictive power (P = 0.040). Furthermore, the CURIO was able to further refine prognosis, separating good versus poor prognosis subgroups in luminal A, luminal B, and triple-negative breast cancer intrinsic subtypes. In breast cancer, the CURIO provides additional prognostic information besides the routinely utilized diagnostic approaches and factors.


Assessing the feasibility, acceptability and potential effectiveness of an integrated approach to smoking cessation for new and expectant fathers: The Dads in Gear study protocol.
Evidence related to the effects of tobacco exposure in pregnancy and on infant and child health have focused on women's smoking cessation. Less often addressed is men's smoking, which when continued in fatherhood, reduces the chances of female partners' cessation and can negatively impact children's health as well as men's health. Dads in Gear (DIG) is an innovative program designed specifically for new fathers who want to reduce and quit smoking that includes three components: smoking cessation, fathering, and physical activity. The over-arching purpose of this study is to evaluate the feasibility of the DIG program and provide estimates of program efficacy. The purpose of this article is to describe the rationale and protocol for evaluating the DIG program's feasibility, acceptability and potential effectiveness. Using a prospective, non-comparative design, the DIG program will be implemented and evaluated in six communities. The program will be offered by trained facilitators to fathers who currently smoke and want to quit. The RE-AIM framework will guide the evaluation. Open-ended questions in participant surveys, and semi-structured interviews and weekly telephone de-briefs with facilitators will provide data for a process evaluation. Estimates of effectiveness include smoking behavior, fathering and physical activity measures at baseline, end of program, and 3-month follow up. The DIG program could support positive changes with respect to smoking cessation, physical activity and overall health for men. These effects could also promote family health. The program might also provide an effective model for engaging men in other health behavior change.


Eye Gaze Patterns Associated with Aggressive Tendencies in Adolescence.
Social information processing theory hypothesizes that aggressive children pay more attention to cues of hostility and threat in others' behavior, consequently leading to over-interpretation of others' behavior as hostile. While there is abundant evidence of aggressive children demonstrating hostile attribution biases, less well documented is whether such biases stem from over-attendance and hypersensitivity to hostile cues in social situations. Over-attendance to hostile cues would be typified by deviations at any stage of the multi-stage process of social information processing models. While deviations at later stages in social information processing models are associated with aggressive behavior in children, the initial step of encoding has historically been difficult to empirically measure, being a low level automatic process unsuitable for self-report. We employed eye-tracking methodologies to better understand the visual encoding of such social information. Eye movements of ten 13-18 year-old children referred from clinical and non-clinical populations were recorded in real time while the children viewed scenarios varying between hostile, non-hostile and ambiguous social provocation. In addition, the children completed a brief measure of risk of aggression. Aggressive children did attend more to the social scenarios with hostile cues, in particular attending longest to those hostile scenarios where the actor in the scenario had a congruent emotional response. These findings corroborate social information processing theory and the traditional bottom-up processing hypotheses that aggressive behavior relates to increased attention to hostile cues.


Mesoporous silica-based hybrid materials for bone-specific drug delivery.
A mesoporous silica-based drug delivery device potentially useful for bone-specific drug delivery has been designed, developed and characterized starting from MSU-type mesoporous silica. The proposed system consists of a mesoporous silica nanoparticles (MSN) based vehicle, presenting alendronate as a targeting functionality for bone tissue while ibuprofen is used as a model molecule for the drugs to be delivered. The particles are functionalized on the external surface using a propionitrile derivative that is successively hydrolyzed to a carboxylic group. Alendronate, one of the most used member of the diphosphonate drug class, is electrostatically bonded to the external carboxyl functionalities of mesoporous silica. The obtained material has been characterized by powder X-ray diffraction, N2 adsorption-desorption porosimetry, UV-vis spectrophotometry, FT-IR spectrometry and MAS-NMR 13C and 29Si. Hydroxyapatite, which simulates the bone matrix, has been synthesized with the aim of testing the targeting activity of the obtained device. In a separate test, the MSNs have been loaded with ibuprofen, a non-steroidal anti-inflammatory drug (NSAID), and its release has been determined under neutral conditions by HPLC. Moreover, biological tests were carried out. The tested devices did not show any toxicity towards normal cells, confirming their high biocompatibility and the lack of off-target effects.


A Computationally Efficient Online/Offline Signature Scheme for Underwater Wireless Sensor Networks.
Underwater wireless sensor networks (UWSNs) have emerged as the most widely used wireless network infrastructure in many applications. Sensing nodes are frequently deployed in hostile aquatic environments in order to collect data on resources that are severely limited in terms of transmission time and bandwidth. Since underwater information is very sensitive and unique, the authentication of users is very important to access the data and information. UWSNs have unique communication and computation needs that are not met by the existing digital signature techniques. As a result, a lightweight signature scheme is required to meet the communication and computation requirements. In this research, we present a Certificateless Online/Offline Signature (COOS) mechanism for UWSNs. The proposed scheme is based on the concept of a hyperelliptic curves cryptosystem, which offers the same degree of security as RSA, bilinear pairing, and elliptic curve cryptosystems (ECC) but with a smaller key size. In addition, the proposed scheme was proven secure in the random oracle model under the hyperelliptic curve discrete logarithm problem. A security analysis was also carried out, as well as comparisons with appropriate current online/offline signature schemes. The comparison demonstrated that the proposed scheme is superior to the existing schemes in terms of both security and efficiency. Additionally, we also employed the fuzzy-based Evaluation-based Distance from Average Solutions (EDAS) technique to demonstrate the effectiveness of the proposed scheme.


Is there a relationship between hand grip strength and knee osteoarthritis in terms of radiological and functional findings in female patients?
The aim of this study was to assess the presence of the relationship between hand grip strength (HGS) and radiological and functional signs of knee osteoarthritis (KOA). Between March 2019 and January 2020, a total of 64 female patients (mean age: 63.4±8.8 years; range, 50 to 80 years) with bilateral chronic knee pain who were diagnosed with KOA using radiological and clinical findings according to the 2019 American College of Rheumatology guidelines were included in the study. Patient demographics, body mass index (BMI) values, and hand dominance were recorded. The Western Ontario and McMaster Universities Arthritis Index (WOMAC) and Lower Extremity Functional Scale (LEFS) were used for the assessment of KOA functionality. The Kellgren-Lawrence (KL) grading system was used for the radiological assessment of KOA. Electronic dynamometer was used for HGS measurement and values were normalized according to BMI results. In the assessment of relationship between HGS and functionality of KOA, a negative, moderate correlation was found between the WOMAC and HGS-Dominant (D) (r=0.312, p<0.05) and HGS-Non-Dominant (ND) (r=0.391, p<0.01). In addition, a positive, moderate correlation was found between the LEFS and HGS-D (r=0.344, p<0.01) and HGS-ND (r=0.371, p<0.01). There was a weak, negative correlation between the HGS-ND, KL-D (r=0.256, p<0.05) and KL-ND (r=0.283, p<0.05), while no significant correlation was found between the HGS-D and KL. Our study results show that HGS-ND is associated with KOA radiologically and functionally. The HGS should be added in the WOMAC and LEFS scales in the functional assessment of KOA and new scales including HGS assessment should be developed.


Gene susceptibility in Iranian asthmatic patients: a narrative review.
As environmental factors are important in the development of asthma, genetic factors could have a critical role in the expression of the disease. Hence, we carried out a systematic review to assess the susceptible genes for asthma in Iranian population. We conducted a literature search by using the electronic database PubMed, Biological Abstracts Web of Science, Current Contents Connect, Cinahl, ScienceDirect, Scopus, IranMedex, and Scientific Information Database to identify articles that evaluated the association between genetic variants and the risk of asthma in Iranian population (until April 30, 2012). The search terms were used include: Asthma and gene in combination with Iran for international database. The following criteria were used for selecting literatures in this review: The study should evaluate the association between gene polymorphism and risk of asthma in Iranian population, and the study should be a case-control design with normal subject as ac control group that published in a journal. Finally, 14 case-control studies were extracted from local and international database. In this study, we reviewed 38 polymorphisms in 19 genes. Polymorphism in interleukin-13 (IL-13), IL-10, IL-1, IL-2, IL-12, E-Selectin, S128R and Exon 9 Vitamin D Receptor were susceptible for asthma and polymorphism in chemokine receptor 5, transforming growth factor-a (TGF-a), Intron 8 of the Vitamin D Receptor, angiotensin-converting enzyme gene, IL-6 and interferon-c were not susceptible for asthma in Iranian population. Polymorphism in IL-4, tumor necrosis factor-α and TGF-b had inconsistent findings. This systematic review indicated that three polymorphisms (IL-13, IL-10, and IL-1) are associated with risk of asthma in Iranian population.


Copper(I)-catalyzed [3+1] cycloaddition of alkenyldiazoacetates and iminoiodinanes: easy access to substituted 2-azetines.
The copper(I)-catalyzed reaction of alkenyldiazoacetates and iminoiodinanes affords functionalized azetine derivatives. This process is consistent with the formation of an aziridinyldiazoacetate intermediate, which gives rise to the four-membered heterocycles by metal-catalyzed ring expansion. The resulting azetine structure is a direct precursor of azeditine-2-carboxylic acid derivatives (EWG = electron-withdrawing group).


Access to the spleen microenvironment through lymph shows local cytokine production, increased cell flux, and altered signaling of immune cells during lipopolysaccharide-induced acute inflammation.
The spleen is involved in fluid volume regulation, immune responses, and hematopoiesis. Yet, the composition of the fluid phase within the spleen microenviroment, the migratory routes of lymphocytes as well as the splenic response to bacterial endotoxin is incomplete. To address these issues, we isolated postnodal lymph in rats by cannulating an efferent lymphatic draining the spleen, and assessed the secretion of signaling substances during a septic response induced by LPS. Spleen lymph flow increased 8-fold after LPS exposure. The spleen exhibited a permeable microvasculature with low sieving of macromolecules that was absent after exposure to LPS. Furthermore, after LPS exposure the spleen contributed significantly to the production of pro- and anti-inflammatory cytokines, and experiments in splenectomized rats suggested it may induce a protracted inflammation because of a dominant role in IL-6 production. A significant amount of lymphocytes exited via lymphatics draining the spleen in control rats. LPS-induced inflammation resulted in increased T cell and reduced B cell subset fractions, and gave a significant increase in CD4(+) and CD8(+) subset T cell efflux and a reduced B cell efflux in spleen lymph. Exposure of leukocytes to the spleen microenvironment affected their signaling status, and by phosphorylation specific flow cytometry we could identify STAT3 and CREB as important mediators in the cellular signaling occurring during endotoxemia. We conclude that analysis of spleen lymph may unravel immune cell migration patterns and local signaling, and immune cells exit via lymph having acquired specific activation signatures after exposure to the spleen microenvironment.


Comparative genomic hybridization analysis of radiation-associated and sporadic meningiomas.
Ionizing irradiation to the skull is a known risk factor for meningioma development. To gain insight into the molecular mechanisms that underlie radiation-associated meningioma (RAM), we characterized the somatic genetic alterations in 16 RAMs by using comparative genomic hybridization and compared the pattern of alterations with 17 nonradiation-associated meningiomas (non-RAM). Most tumors (29/33;87.9%) displayed at least one DNA copy number alteration, and 11 out of 33 (33%) exhibited four or more changes. The mean number of DNA copy number changes was similar in RAMs (2.4+/-1.9) and in non-RAMs (2.5+/-1.9). The most common DNA losses were noted in chromosome 22 (56.2% in RAM, and 47% in non-RAM) and chromosome 1 (37.5% in RAM and 35.3% in non-RAM), with no significant differences between the two groups. Noteworthy, gain in DNA copy number of chromosomes 8 and 12 was detected in two RAM tumors only. In conclusion, no significant differences were noted between RAMs and non-RAMs regarding the number of genetic changes and the extent and frequency of chromosomes 1 and 22 losses. These preliminary data suggest that the tumorogenic pathways of meningioma formation are similar, regardless of previous skull irradiation.


Timing of the introduction of complementary feeding and risk of childhood obesity: a systematic review.
The World Health Organisation recommends exclusive breastfeeding until 6 months of age and continued breastfeeding until 2 years of age or beyond. Appropriate complementary foods should be introduced in a timely fashion, beginning when the infant is 6 months old. In developing countries, early or inappropriate complementary feeding may lead to malnutrition and poor growth, but in countries such as the United Kingdom and United States of America, where obesity is a greater public health concern than malnutrition, the relationship to growth is unclear. We conducted a systematic review of the literature that investigated the relationship between the timing of the introduction of complementary feeding and overweight or obesity during childhood. Electronic databases were searched from inception until 30 September 2012 using specified keywords. Following the application of strict inclusion/exclusion criteria, 23 studies were identified and reviewed by two independent reviewers. Data were extracted and aspects of quality were assessed using an adapted Newcastle-Ottawa scale. Twenty-one of the studies considered the relationship between the time at which complementary foods were introduced and childhood body mass index (BMI), of which five found that introducing complementary foods at <3 months (two studies), 4 months (2 studies) or 20 weeks (one study) was associated with a higher BMI in childhood. Seven of the studies considered the association between complementary feeding and body composition but only one study reported an increase in the percentage of body fat among children given complementary foods before 15 weeks of age. We conclude that there is no clear association between the timing of the introduction of complementary foods and childhood overweight or obesity, but some evidence suggests that very early introduction (at or before 4 months), rather than at 4-6 months or >6 months, may increase the risk of childhood overweight.


Standardizing assessment practices of undergraduate medical competencies across medical schools: challenges, opportunities and lessons learned from a consortium of medical schools in Uganda.
Health professions education is gradually moving away from the more traditional approaches to new innovative ways of training aimed at producing professionals with the necessary competencies to address the community health needs. In response to these emerging trends, Medical Education for Equitable Services to All Ugandans (MESAU), a consortium of Ugandan medical schools developed key competencies desirable of graduates and successfully implemented Competency Based Education (CBE) for undergraduate medical students. To examine the current situation and establish whether assessment methods of the competencies are standardized across MESAU schools as well as establish the challenges, opportunities and lessons learned from the MESAU consortium. It was a cross-sectional descriptive study involving faculty of the medical schools in Uganda. Data was collected using focus group discussions and document reviews. Findings were presented in form of themes. Although the MESAU schools have implemented the developed competencies within their curricular, the assessment methods are still not standardized with each institution having its own assessment procedures. Lack of knowledge and skills regarding assessment of the competencies was evident amongst the faculty. The fear for change amongst lecturers was also noted as a major challenge. However, the institutional collaboration created while developing competencies was identified as key strength. Findings demonstrated that despite having common competencies, there is no standardized assessment blue print applicable to all MESAU schools. Continued collaboration and faculty development in assessment is strongly recommended.


Combined immunostimulatory monoclonal antibodies extend survival in an aggressive transgenic hepatocellular carcinoma mouse model.
Immunostimulatory monoclonal antibodies (ISmAb) that unleash antitumor immune responses are showing efficacy in cancer clinical trials. Anti-B7-H1 (PD-L1) monoclonal antibodies (mAb) block a critical inhibitory pathway in T cells, whereas anti-CD137 and OX40 mAbs provide T-cell costimulation. A combination of these ISmAbs (anti-CD137 + anti-OX40 + anti-B7-H1) was tested using a transgenic mouse model of multifocal and rapidly progressing hepatocellular carcinoma, in which c-myc drives transformation and cytosolic ovalbumin (OVA) is expressed in tumor cells as a model antigen. Flow-cytometry and immunohistochemistry were used to quantify tumor-infiltrating lymphocytes (TIL) elicited by treatment and assess their activation status and cytolytic potential. Tolerance induction and its prevention/reversal by treatment with the combination of ISmAbs were revealed by in vivo killing assays. The triple combination of ISmAbs extended survival of mice bearing hepatocellular carcinomas in a CD8-dependent fashion and synergized with adoptive T-cell therapy using activated OVA-specific TCR-transgenic OT-1 and OT-2 lymphocytes. Mice undergoing therapy showed clear increases in tumor infiltration by activated and blastic CD8(+) and CD4(+) T lymphocytes containing perforin/granzyme B and expressing the ISmAb-targeted receptors on their surface. The triple combination of ISmAbs did not result in enhanced OVA-specific cytotoxic T lymphocyte (CTL) activity but other antigens expressed by cell lines derived from such hepatocellular carcinomas were recognized by endogenous TILs. Adoptively transferred OVA-specific OT-1 lymphocytes into tumor-bearing mice were rendered tolerant, unless given the triple mAb therapy. Extension of survival and dense T-cell infiltrates emphasize the translational potential of combinational immunotherapy strategies for hepatocellular carcinoma. Clin Cancer Res; 19(22); 6151-62. ©2013 AACR.


Uncovering the Role of RNA-Binding Protein hnRNP K in B-Cell Lymphomas.
Heterogeneous nuclear ribonucleoprotein K (hnRNP K) is an RNA-binding protein that is aberrantly expressed in cancers. We and others have previously shown that reduced hnRNP K expression downmodulates tumor-suppressive programs. However, overexpression of hnRNP K is the more commonly observed clinical phenomenon, yet its functional consequences and clinical significance remain unknown. Clinical implications of hnRNP K overexpression were examined through immunohistochemistry on samples from patients with diffuse large B-cell lymphoma who did not harbor MYC alterations (n = 75). A novel transgenic mouse model that overexpresses hnRNP K specifically in B cells was generated to directly examine the role of hnRNP K overexpression in mice (three transgenic lines). Molecular consequences of hnRNP K overexpression were determined through proteomics, formaldehyde-RNA-immunoprecipitation sequencing, and biochemical assays. Therapeutic response to BET-bromodomain inhibition in the context of hnRNP K overexpression was evaluated in vitro and in vivo (n = 3 per group). All statistical tests were two-sided. hnRNP K is overexpressed in diffuse large B-cell lymphoma patients without MYC genomic alterations. This overexpression is associated with dismal overall survival and progression-free survival (P < .001). Overexpression of hnRNP K in transgenic mice resulted in the development of lymphomas and reduced survival (P < .001 for all transgenic lines; Line 171[n = 30]: hazard ratio [HR] = 64.23, 95% confidence interval [CI] = 26.1 to 158.0; Line 173 [n = 31]: HR = 25.27, 95% CI = 10.3 to 62.1; Line 177 [n = 25]: HR = 119.5, 95% CI = 42.7 to 334.2, compared with wild-type mice). Clinical samples, mouse models, global screening assays, and biochemical studies revealed that hnRNP K's oncogenic potential stems from its ability to posttranscriptionally and translationally regulate MYC. Consequently, Hnrnpk overexpression renders cells sensitive to BET-bromodomain-inhibition in both in vitro and transplantation models, which represents a strategy for mitigating hnRNP K-mediated c-Myc activation in patients. Our findings indicate that hnRNP K is a bona fide oncogene when overexpressed and represents a novel mechanism for c-Myc activation in the absence of MYC lesions.


A unique neuropsychophysiological approach to objectify emotion (dys)regulation in healthy older adults during the COVID-19 pandemic.
The response of older people to the COVID-19 pandemic has attracted much attention as they are at increased risk of adverse outcomes. A longitudinal study has shown that improvement in global cognitive, executive and language functioning in healthy older adults enrolled at the University of the Third Age appears to play a protective role against emotional dysregulation and mood changes during the pandemic. To date, no study has examined emotional dysregulation through COVID-19-related images using facial electromyographic recordings in healthy older adults. Therefore, we aimed to analyze the relationships between zygomaticus and corrugator reactivity, neuropsychological measures, and the affective dimensions of arousal, dominance, and valence. The results showed an unexpected association between higher zygomaticus activity and higher levels of apathy, depression, and anxiety. In contrast, increased contracture of the corrugator was associated with poorer performance on cognitive tests (global cognition, memory, executive functions) and physical status, i.e., walking speed. These results are consistent with the reappraisal of emotional stimuli in response to the challenges of the pandemic. Interestingly, COVID-19-related stimuli triggered the activation of bottom-up affectivity strategies associated with higher mood levels and interacted with top-down factors that play an important role in the dysregulation of cognitive control.


Mourning the dead, mourning the disappeared: the enigma of the absent-presence.
Freud's interest in the impact of death on the living goes back further than Mourning and Melancholia (1917e, [1915]). In Totem and Taboo (1912-13) Freud noted the ambivalence of the emotions we experience in relation to the dead. In this paper, I focus on Mourning and Melancholia as a landmark in the understanding of both the normal and psychopathological aspects of mourning and depressive processes in human beings. Mourning and Melancholia bridges Freud's first and second topographic theories of the psychic apparatus and constitutes for many authors the foundation of his theory of internal object relations. With this psychoanalytic understanding of mourning as a framework, I discuss 'special mourning processes,' such as the those confronted by psychoanalysts in Argentina when treating the relatives of thousands of people who were 'disappeared' by the military dictatorship in the 1970s; they are 'special' in the sense that the external reality [which] constitutes the starting point of the psychic mourning process, as described by Freud, is absent. I argue that the 'absent-presence' of the body as an enigmatic message initiates a special mourning process that bears certain characteristics of, and is isomorphic to, Laplanche's seduction theory.


Cytokine Adsorption in Critically Ill COVID-19 Patients, a Case-Control Study.
Aim: New coronavirus disease (COVID-19) has become an international emergency. As many of the intensive care unit (ICU) patients with the disease also present multiple organ failure, blood purification techniques might be a good choice in their treatment. In this study we aimed to investigate the role of cytokine removal in COVID-19 patients managed in ICUs. Methods: For this case-control study we have investigated the role of the cytokine removal by means of two resin membranes (HA330 and Mediasorb) in COVID-19 patients managed in ICUs. Particularly, we investigated the overtime variation in clinical severity scores, laboratory variables, and effects on hospital and ICU stay and mortality. Results: Seventy-two patients have been evaluated, of which half constituted Cytokine Filtration (CF) Group, and other half the Case-Control (CC) Group. Mortality was 55.6% and 50% in CF and CC groups, respectively. In the CF Group, there was decrease in C-reactive protein (CRP) and fibrinogen levels measured at the end of cytokine adsorption; lymphocyte count and ratio were increased, whereas neutrophile ratio was decreased. There were no differences between the groups regarding other laboratory variables, SOFA scores and vasopressor uses. Conclusions: We have demonstrated decrease in CRP, fibrinogen and increase in lymphocyte count in the patients having cytokine adsorption, but there was no clinical reflection of these benefits, and no decrease in mortality as well. Even though there is physio-pathologic rationale to use cytokine adsorption techniques for immunomodulation in critically ill COVID-19 patients, it is early to make strong suggestions about their benefits.


Proton NMR relaxation times in the peripheral blood of cancer patients.
The proton spin lattice relaxation time (T1) of serum and leucocytes of cancer patients and normal volunteers was measured using pulsed NMR techniques. There was no statistically significant difference in the serum T1 values of cancer patients relative to normal. An increase in T1 relative to normal values was detected in the white blood cells of patients with active leukaemia. In these patients T1 fell to normal levels after the initiation of treatment. The variation of leucocyte T1 with the course of the disease for five patients having leukaemia is presented.


Revision and restandardization of the Conners Teacher Rating Scale (CTRS-R): factor structure, reliability, and criterion validity.
The Conners Teacher Rating Scale (CTRS) is a commonly used research and clinical tool for assessing children's behavior in the classroom. The present study introduces the revised CTRS (CTRS-R) which improves on the original CTRS by (1) establishing normative data from a large, representative North American sample, (2) deriving a factor structure using advanced statistical techniques, and (3) updating the item content to reflect current conceptualizations of childhood disorders. Using confirmatory factor analysis, a six-factor structure was found which includes Hyperactivity-Impulsivity, Perfectionism, Inattention/Cognitive Problems, Social Problems, Oppositionality, and Anxious/Shy factors. The reliability of the scale, as measured by test-retest correlations and internal consistency, is generally satisfactory. Using all of the scale factors to discriminate between attention deficit hyperactivity disordered and normal children, 85 percent of children were correctly classified, supporting the validity of the scale and indicating excellent clinical utility. Similarities and differences between the original CTRS factor structure and the CTRS-R factor structure are discussed.


[Changes in neurohumoral factors of various tissues in patients with breast cancer].
Such neurohumoral factors of antitumor resistance regulation as catecholamines, corticosteroids and cholinergic mechanisms were studied in tissues of breast cancer patients. Tissue adjacent to tumor revealed elevated concentrations of 11-hydroxycorticosteroids and adrenaline matched by an increased release of norepinephrine which greatly depended on weakened cholinergic influences. These disturbances were pronounced in stages I and II tumors and, particularly, in infiltrating scirrhous cancer. Disbalance in neurohumoral factor distribution between normal and altered tissues may boost tumor recurrence and metastatic spread.


Flow cytometric analysis of human lung cancer. Correlation with histologic type and stage.
DNA ploidy and cell-cycle characteristics of 65 operable lung cancers (41 adenocarcinomas, 19 epidermoid carcinomas, 3 large-cell carcinomas and 2 small-cell carcinomas) were analyzed using flow cytometry. Eighty percent of the tumors were aneuploid. The mean DNA index was lower in epidermoid than in adenocarcinoma. In adenocarcinoma, a low DNA index was correlated with early-stage disease; no correlation between DNA index and stage was observed in the other cell types. The %S-phase cells was highest in two cases of undifferentiated large-cell carcinoma and lowest in adenocarcinoma. The RNA index was increased approximately two-fold in all cell types. Longer follow-ups will be required to establish any correlation between the cell kinetic measurements reported here and survival times.


The role of mechanical stresses in angiogenesis.
Angiogenesis is the formation of new capillary blood vessels from preexisting vessels. It is involved in many normal and diseased conditions, as well as in the application of tissue-engineered products. There has been extensive effort made to develop strategies for controlling pathological angiogenesis and for promoting vascularization in biomedical engineering applications. Central to advancing these strategies is a mechanistic understanding of the angiogenic process. Angiogenesis is tightly regulated by local tissue environmental factors, including soluble molecules, extracellular matrices, cell-cell interactions, and diverse mechanical forces. Great advances have been made in identifying the biochemical factors and intracellular signaling pathways that mediate the control of angiogenesis. This review focuses on work that explores the biophysical aspect of angiogenesis regulation. Specifically, we discuss the role of cell-generated forces, counterforces from the extracellular matrix, and mechanical forces associated with blood flow and extravascular tissue activity in the regulation of angiogenesis. Because angiogenesis occurs in a mechanically dynamic environment, future investigations should aim at understanding how cells integrate chemical and mechanical signals so that a rational approach to controlling angiogenesis will become possible. In this regard, computational models that incorporate multiple epigenetic factors to predict capillary patterning will be useful.


Overexpression of p27Kip1 inhibits the growth of both normal and transformed human mammary epithelial cells.
We previously reported increased expression of p27Kip1 in a series of human breast cancer cell lines, as compared to cell lines established from normal mammary epithelial cells. These data were surprising because this protein exerts a growth-inhibitory function in normal cells, and p27Kip1 has been proposed as a candidate tumor suppressor gene. A possible explanation for the paradoxical increase in p27Kip1 in the breast cancer cell lines is that they had become refractory to the inhibitory effects of this protein. To address this question, here, we transfected the MCF7 breast cancer cell line and the MCF10F nontumorigenic mammary epithelial cell line with a vector containing the p27Kip1 cDNA to obtain derivatives that express increased levels of p27Kip1. The increased expression of p27Kip1 in both of these cell lines was associated with lengthening of the G1 phase, an increase in the doubling time, a decreased saturation density, and a decreased plating efficiency. In the MCF7 cells, anchorage-independent growth and in vivo tumorigenicity were also suppressed. These effects were associated with decreased cyclin E-associated in vitro kinase activity in both cell lines. The increased expression of p27Kip1 was associated with a decreased level of expression of cyclin D1 in the MCF10F cells but an increased level of the cyclin D1 protein in the MCF7 cell line. Both derivatives expressed slightly increased levels of the cyclin E protein. Thus, breast cancer cells are still responsive to p27Kip1-mediated inhibition of cell growth despite the high basal level of this protein. These results suggest that therapeutic strategies that further increase the level of expression of p27Kip1 or mimic its activity might be useful in cancer therapy.


Optimizing therapy of bloodstream infection due to extended-spectrum β-lactamase-producing Enterobacteriaceae.
Infections due to extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) are increasing worldwide. Carbapenems are usually regarded as the antibiotics of choice for the treatment of serious ESBL infections. However, because of the alarming emergence or carbapenem resistance, interest in effective alternatives has emerged. The present review summarizes the findings published on the antibiotics currently available for treatment of patients with an ESBL-E bloodstream infection (BSI). Meropenem and imipenem are the drugs recommended for treatment of ESBL BSIs in critically ill patients, and in infections with high bacterial loads or elevated β-lactam minimum inhibitory concentrations. Ertapenem should be reserved for patients with less severe presentations, and should be used at high doses. In milder presentations or BSIs from low-risk sources, other carbapenem-sparing alternatives could be considered: cephamycins, fluoroquinolones, and particularly a β-lactam/β-lactam inhibitor combination (particularly piperacillin/tazobactam). Optimized dosing of piperacillin/tazobactam is recommended (high doses and extended infusion). There are few data on the use of the promising newly available drugs (e.g. ceftolozane/tazobactam, ceftazidime/avibactam, cefiderocol, and plazomicin), and it seems reasonable to reserve them as last-resort drugs. Carbapenems should be used in patients with serious infections; alternatives could be used individually, particularly for definitive treatment of patients with milder presentations.


Gut-microbiota interactions in non-mammals: what can we learn from Drosophila?
Millions of people suffer from inflammatory diseases of the intestine, some of them potentiating gastrointestinal cancer. These gut-associated pathologies arise from imbalanced interactions between the host gut epithelia and resident or ingested microbes, interactions that are still poorly understood at the molecular level. Drosophila has been a very powerful model to study development and diseases. Its relatively simple tissue organization and sophisticated genetics are some of the advantages of using it as an experimental model to dissect gut-microbe interactions. Recent progress made in various research fields such as Drosophila microbiota composition, gut epithelium structure or gut immune reactions led us to believe that Drosophila is becoming an ad hoc model system to dissect the mechanisms that cooperate to maintain intestinal homeostasis in higher eukaryotes. It further may help us understand how an alteration of these finely tuned processes precipitates the inflammatory processes found in some inflammatory bowel diseases.


Maternal serum interleukin 6 levels and fetomaternal outcomes in women with preterm premature rupture of membranes in Lagos, South-western Nigeria.
This study was done to assess the relationship between maternal serum IL-6 levels and fetomaternal outcome following PPROM. This was a prospective cohort study comprising 45 cases of PPROM and 45 controls of similar age, parity, and gestational age. Five milliliters of maternal serum was collected after obtaining informed consent. They were followed up till delivery and records of the delivery and neonatal outcomes were obtained. Serum IL-6 levels were determined by standard enzyme-linked immunosorbent assay [ELISA]. PPROM patients were categorized into two groups using a threshold of 14 pg/ml. Chi-square (χ2) test was used to compare categorical outcomes. p values of < 0.05 were taken as significant. The mean serum IL-6 level for the women with PPROM was (20.2 ± 11.0 pg/ml), which was significantly greater than for the control subjects (13.9 ± 5.8 pg/ml, p < 0.001). Fetomaternal outcomes were all worse in those with IL-6 ≥ 14 pg/ml. Nevertheless, only the difference in early neonatal deaths was statistically significant. Measurement of maternal serum IL-6 can help to indicate hostile intrauterine environments to the fetus as well as identify patients who may benefit from pregnancy prolongation or intervention.


Efficacy of standardised manual therapy and home exercise programme for chronic rotator cuff disease: randomised placebo controlled trial.
To investigate the efficacy of a programme of manual therapy and exercise treatment compared with placebo treatment delivered by physiotherapists for people with chronic rotator cuff disease. Randomised, participant and single assessor blinded, placebo controlled trial. Metropolitan region of Melbourne, Victoria, Australia. 120 participants with chronic (>3 months) rotator cuff disease recruited through medical practitioners and from the community. The active treatment comprised a manual therapy and home exercise programme; the placebo treatment comprised inactive ultrasound therapy and application of an inert gel. Participants in both groups received 10 sessions of individual standardised treatment over 10 weeks. For the following 12 weeks, the active group continued the home exercise programme and the placebo group received no treatment. The primary outcomes were pain and function measured by the shoulder pain and disability index, average pain on movement measured on an 11 point numerical rating scale, and participants' perceived global rating of overall change. 112 (93%) participants completed the 22 week trial. At 11 weeks no difference was found between groups for change in shoulder pain and disability index (3.6, 95% confidence interval -2.1 to 9.4) or change in pain (0.7, -0.1 to 1.5); both groups showed significant improvements. More participants in the active group reported a successful outcome (defined as "much better"), although the difference was not statistically significant: 42% (24/57) of active participants and 30% (18/61) of placebo participants (relative risk 1.43, 0.87 to 2.34). The active group showed a significantly greater improvement in shoulder pain and disability index than did the placebo group at 22 weeks (between group difference 7.1, 0.3 to 13.9), although no significant difference existed between groups for change in pain (0.9, -0.03 to 1.7) or for the percentage of participants reporting a successful treatment outcome (relative risk 1.39, 0.94 to 2.03). Several secondary outcomes favoured the active group, including shoulder pain and disability index function score, muscle strength, interference with activity, and quality of life. A standardised programme of manual therapy and home exercise did not confer additional immediate benefits for pain and function compared with a realistic placebo treatment that controlled for therapists' contact in middle aged to older adults with chronic rotator cuff disease. However, greater improvements were apparent at follow-up, particularly in shoulder function and strength, suggesting that benefits with active treatment take longer to manifest. Clinical trials NCT00415441.


A Systematic Study of Unsaturation in Lipid Nanoparticles Leads to Improved mRNA Transfection In Vivo.
Lipid nanoparticles (LNPs) represent the leading concept for mRNA delivery. Unsaturated lipids play important roles in nature with potential for mRNA therapeutics, but are difficult to access through chemical synthesis. To systematically study the role of unsaturation, modular reactions were utilized to access a library of 91 amino lipids, enabled by the synthesis of unsaturated thiols. An ionizable lipid series (4A3) emerged from in vitro and in vivo screening, where the 4A3 core with a citronellol-based (Cit) periphery emerged as best. We studied the interaction between LNPs and a model endosomal membrane where 4A3-Cit demonstrated superior lipid fusion over saturated lipids, suggesting its unsaturated tail promotes endosomal escape. Furthermore, 4A3-Cit significantly improved mRNA delivery efficacy in vivo through Selective ORgan Targeting (SORT), resulting in 18-fold increased protein expression over parent LNPs. These findings provide insight into how lipid unsaturation promotes mRNA delivery and demonstrate how lipid mixing can enhance efficacy.


JMFit: A SAS Macro for Joint Models of Longitudinal and Survival Data.
Joint models for longitudinal and survival data now have a long history of being used in clinical trials or other studies in which the goal is to assess a treatment effect while accounting for a longitudinal biomarker such as patient-reported outcomes or immune responses. Although software has been developed for fitting the joint model, no software packages are currently available for simultaneously fitting the joint model and assessing the fit of the longitudinal component and the survival component of the model separately as well as the contribution of the longitudinal data to the fit of the survival model. To fulfill this need, we develop a SAS macro, called JMFit. JMFit implements a variety of popular joint models and provides several model assessment measures including the decomposition of AIC and BIC as well as ΔAIC and ΔBIC recently developed in Zhang et al. (2014). Examples with real and simulated data are provided to illustrate the use of JMFit.


Pain control in thoracic oncology.
This review of pain management in lung cancer is based on the presentation of four cases of thoracic oncology patients with pain at various stages of their disease. The approach will be multidisciplinary, involving a thoracic oncologist, radiologist, thoracic and orthopaedic spine surgeon, radiation therapist, pain medicine specialist, and palliative care specialist. This multispecialty approach to the management of different painful presentations in thoracic oncology will demonstrate the complexity of each case and the improved patient outcomes which result from the involvement of different disciplines working in concert.In the USA, Europe and other countries, palliative care specialists often become rapidly involved in the management of these patients, coordinating social care and providing psychological support.Thoracic and orthopaedic spine subspecialists provide surgical methods to control tumour invasion, and improve quality of life and preservation of function in settings of even diffuse metastatic disease. Similarly, thoracic oncology and radiation therapists utilise both therapeutic and palliative chemotherapeutic and radiation therapy regimens to prolong and improve quality of life.The pain medicine specialist can, in addition to medication management, offer a variety of interventional approaches including unique drug delivery systems such as epidural analgesia, regional anaesthesia techniques, and intrathecal pumps, as well as neuromodulation techniques and neurolytic or neuroablative procedures.In the USA, these specialists complete an additional fellowship year in pain medicine following the completion of an anaesthesiology, physical medicine and rehabilitation, neurology or psychiatry residency. These programmes are accredited by the Accreditation Council for Graduate Medical Education, or ACGME (www.acgme.org).


A phenomenon-oriented strategy in depression research.
This paper discusses the presence of important uninvestigated assumptions in every aspect of depression research. It is suggested that a general phenomenon-oriented strategy might reduce the number of assumptions, if pursued in accordance with the requirements of natural science, which aims at maximizing the accuracy of our ideas about phenomena. Possible consequences of this strategy are: (1) the provisional selection of single, well-established, target and criterion variables, together with comparison variables, all suggested by special investigations and previous research results; (2) maximizing the phenomenological validity of current observation procedures, in the light of results of research into questionnaire and interview methods; (3) inclusion of hitherto neglected, but scientifically legitimate, individual-centred, experimental and longitudinal specific strategies: the strategies of choice for the investigation of psychological processes; (4) invention of tactics to facilitate the ethically acceptable use of scientific method in investigations of clinically relevant psychological phenomena; (5) explication and eventual testing of all assumptions; (6) focus upon the understanding, prediction and control of well-established clinically relevant phenomena, with theory playing a secondary role of facilitating systematic investigation; and (7) immediate restriction, and eventual cessation, of the use of current forms of psychiatric diagnosis in fundamental depression research.


Photochromic and photothermal hydrogels derived from natural amino acids and heteropoly acids.
A new class of supramolecular hydrogels have been designed and synthesized via the co-assembly of basic amino acids (AAs) and heteropoly acids (HPAs) under acidic conditions. The formation of gel-like samples is identified using an inverted tube method, rheology, and scanning and transmission electron microscopy. Fourier transform infrared spectroscopy reveals that the structural integrity of the HPAs is maintained during the gelation process. X-ray photoelectron spectroscopy (XPS) and proton nuclear magnetic resonance spectroscopy demonstrate that the anionic HPAs interact with both the protonated α-NH2 and the protonated side groups of the basic amino acids, initiating the preferential growth of one-dimensional nanofibers. These nanofibers bundle and entangle with each other to form extended three-dimensional network structures. The resulting AA/HPA supramolecular hydrogels show clear stereoselectivity of the basic amino acids. With the decreasing enantiomeric excess of the basic amino acids, the gelation propensity of the AA/HPA complexes is found to be depressed. The co-assembled hydrogels show the UV-responsive photochromic behaviour because of the presence of HPAs. The corresponding XPS data confirm that the photochromism of the hydrogels is attributed to the intervalence charge-transfer transition resulting from the reduction of HPAs. Interestingly, the reduced HPAs within the hydrogel matrix can absorb the near-infrared (NIR) light and exhibit photo-thermal conversion properties, which elevates the bulk temperature of the AA/HPA hydrogels and induces the gel-to-sol transition. This study unveils that HPAs have unique capacity to promote the gelation of basic amino acids for the construction of supramolecular soft materials with functional features.


Use of Broken-Symmetry Density Functional Theory To Characterize the IspH Oxidized State: Implications for IspH Mechanism and Inhibition.
With current therapies becoming less efficacious due to increased drug resistance, new inhibitors of both bacterial and malarial targets are desperately needed. The recently discovered methylerythritol phosphate (MEP) pathway for isoprenoid synthesis provides novel targets for the development of such drugs. Particular attention has focused on the IspH protein, the final enzyme in the MEP pathway, which uses its [4Fe-4S] cluster to catalyze the formation of the isoprenoid precursors IPP and DMAPP from HMBPP. IspH catalysis is achieved via a 2e-/2H+ reductive dehydroxylation of HMBPP; the mechanism by which catalysis is achieved, however, is highly controversial. The work presented herein provides the first step in assessing different routes to catalysis by using computational methods. By performing broken-symmetry density functional theory (BS-DFT) calculations that employ both the conductor-like screening solvation model (DFT/COSMO) and a finite-difference Poisson-Boltzmann self-consistent reaction field methodology (DFT/SCRF), we evaluate geometries, energies, and Mössbauer signatures of the different protonation states that may exist in the oxidized state of the IspH catalytic cycle. From DFT/SCRF computations performed on the oxidized state, we find a state where the substrate, HMBPP, coordinates the apical iron in the [4Fe-4S] cluster as an alcohol group (ROH) to be one of two, isoenergetic, lowest-energy states. In this state, the HMBPP pyrophosphate moiety and an adjacent glutamate residue (E126) are both fully deprotonated, making the active site highly anionic. Our findings that this low-energy state also matches the experimental geometry of the active site and that its computed isomer shifts agree with experiment validate the use of the DFT/SCRF method to assess relative energies along the IspH reaction pathway. Additional studies of IspH catalytic intermediates are currently being pursued.


Opiate- and alpha 2-adrenoceptor-induced hyperpolarizations of locus ceruleus neurons in brain slices: reversal by cyclic adenosine 3':5'-monophosphate analogues.
The purpose of this study was to investigate the ionic and second messenger mechanisms underlying the hyperpolarizations induced by the selective alpha 2-adrenoceptor agonist clonidine and the opiate agonist morphine in the locus ceruleus. Intracellular recordings were carried out in rat brain slices, and drugs at known concentrations were administered in the perfusate. The cyclic adenosine 3':5'-monophosphate (cAMP) analogues 8-bromo-cAMP and dibutyryl cAMP, while not altering basal activity, reversed the hyperpolarizations induced by clonidine or morphine. In contrast, administration of the parent compound adenosine failed to affect these responses. These results are consistent with previous biochemical studies suggesting that alpha 2-adrenergic and opiate agonists might signal their actions by reducing intracellular cAMP levels. Under manual voltage clamp, both clonidine and morphine elicited outward currents. The algebraic sum of the individual currents elicited by morphine and clonidine significantly exceeded the actual current elicited by their co-administration. This nonadditivity, as well as the observation that cAMP analogues reverse the morphine- and clonidine-induced hyperpolarizations, suggests that these compounds hyperpolarize locus ceruleus neurons through a shared ionic mechanism the activation of which might be signaled by a decrease in intracellular cAMP.


EEG-based Machine Learning Models to Evaluate Haptic Delay: Should We Label Data Based on Self-Reporting or Physical Stimulation?
Reliable haptic interfaces augment human-computer interaction via simulated tactile and kinesthetic feedback. As haptic technologies advance, user experience evaluation becomes more crucial. Conventionally, self-reporting is used to evaluate haptic experiences; however, it could be inconsistent or imprecise due to human error. A promising alternative is using neurocognitive methods with machine or deep learning models to evaluate the human haptic experience. Machine and deep learning models can be trained on Electroencephalography (EEG) data labeled based on self-report or actual physical stimulation. As the literature lacks a systematic study on which approach is more robust, we develop a visuo-haptic task to answer this question by examining an important haptic experience, namely, haptic delay. EEG is recorded during the experiment, and participants report whether they detected a delay in the haptic modality through self-report. Four machine/deep learning models were trained twice on the EEG data using the two labeling methods. Models trained with labels from the physical stimuli significantly outperformed those trained with self-reporting labels. Although this finding holds true for one particular haptic experience (haptic delay), it cannot be extrapolated to others; rather, it suggests that EEG data labeling plays a prominent role in evaluating the haptic experience through neurocognitive methods.


[Organization and management of clinical-care pathways at the Regina Margherita Hospital in Turin during Covid-19 emergency].
The Covid-19 pandemic significantly increased the workload for the Italian Health Service. There is few information in the literature on the pediatric population and on the management of pediatric hospitals. The aim of this article is to describe the management of healthcare services during Covid-19 emergency in Regina Margherita Children's Hospital. The Regina Margherita Children's Hospital is specialized in the prevention, diagnosis and treatment of pediatric diseases. About 1000 health worker work in this Hospital and 278 hospitalization places are available.


Synchronous elective contralateral mastectomy and immediate bilateral breast reconstruction in women with early-stage breast cancer.
The role of elective contralateral mastectomy (ECM) in women with early-stage breast cancer who elect or require an ipsilateral mastectomy and desire immediate bilateral breast reconstruction (IBR) is an intellectual and emotional dilemma for both patient and physician. In an attempt to clarify the rationale for this approach, we reviewed our experience with ECM and IBR and evaluated operative morbidity, the incidence of occult contralateral breast cancer, and patterns of recurrence. We retrospectively reviewed the records of 155 patients with primary unilateral breast cancer (stage 0, I, or II) and negative findings on physical and mammographic examinations of the contralateral breast who underwent ipsilateral mastectomy and simultaneous ECM with IBR between 1987 and 1995. The median age of the patients was 46 years (range, 25 to 69 years). Clinical stage at diagnosis was stage 0, I, and II in 19.4%, 54.2%, and 26.4% of patients, respectively. Factors likely to influence the use of ECM were family history of breast cancer in first-degree relatives (30%), any family history of breast cancer (56%), difficulty anticipated in contralateral breast surveillance (48%), associated lobular carcinoma in situ (23%), multicentric primary tumor (28%), significant reconstructive issues (14%), and failure of mammographic identification of the primary tumor (16%). Skin-sparing mastectomies were performed in 81% of patients. Overall, 70% of patients underwent reconstruction using autogenous tissue transfer. Reoperations for suspected anastomotic thrombosis were performed in seven patients. Two patients experienced significant partial or complete flap loss. Histopathologic findings in the ECM specimen were as follows: benign, 80% of patients; atypical ductal hyperplasia, 12% of patients; lobular carcinoma in situ, 6.5% of patients; ductal carcinoma in situ, 2.7% of patients; and invasive carcinoma, 1.3% of patients. Eighteen patients (12%) had evidence of locoregional or distant recurrences, with a median follow-up of 3 years. In one patient (0.6%), invasive ductal carcinoma developed on the side of the elective mastectomy. The use of ECM and IBR cannot be justified if the only oncologic criterion considered is the incidence of occult synchronous contralateral disease. However, in a highly selected population of young patients with a difficult clinical or mammographic examination and an increased lifetime risk of developing a second primary tumor, ECM and IBR is a safe approach.


In vitro Antimicrobial Activity of Chlorquinaldol against Microorganisms Responsible for Skin and Soft Tissue Infections: Comparative Evaluation with Gentamicin and Fusidic Acid.
Skin and soft tissue infections (SSTIs) are a major therapeutic challenge for clinicians. The emergence of pathogens with decreased susceptibility to available therapies has become an emerging problem often associated with treatment failure. Hence, there is an urgent need for novel broad-spectrum antimicrobial agents. The purpose of this study was to assess the feasibility of chlorquinaldol as an alternative approach to currently used topical antibiotics for the treatment of skin and soft tissue infections. The activity of chlorquinaldol was investigated against a collection of bacterial isolates responsible for skin infections, including strains resistant to fusidic acid and gentamicin. After determination of MIC and MBC, time-kill experiments were carried out by counting colonies grown after 0, 3, 6, 9, 24, and 48 h of incubation with concentrations equal to ¼×, ½×, 1×, 2×, and 4× MIC of chlorquinaldol, gentamicin, or fusidic acid. Staphylococci resulted the Gram-positives most sensitive to chlorquinaldol, with MIC-values ranging from 0.016 to 0.5 mg/L. A lower activity was observed against Gram-negative bacteria, with 77% of the isolates being inhibited at concentrations ranging from 128 to 512 mg/L. Generally, in time-kill studies, chlorquinaldol showed a bactericidal activity at the higher concentrations (2×, 4× MIC) after 24-48 h of incubation. In conclusion, chlorquinaldol may represent a valuable alternative to conventional topical antibiotics for the treatment of skin and soft tissue infections.


CDH1 regulates E2F1 degradation in response to differentiation signals in keratinocytes.
The E2F1 transcription factor plays key roles in skin homeostasis. In the epidermis, E2F1 expression is essential for normal proliferation of undifferentiated keratinocytes, regeneration after injury and DNA repair following UV radiation-induced photodamage. Abnormal E2F1 expression promotes nonmelanoma skin carcinoma. In addition, E2F1 must be downregulated for proper keratinocyte differentiation, but the relevant mechanisms involved remain poorly understood. We show that differentiation signals induce a series of post-translational modifications in E2F1 that are jointly required for its downregulation. Analysis of the structural determinants that govern these processes revealed a central role for S403 and T433. In particular, substitution of these two amino acid residues with non-phosphorylatable alanine (E2F1 ST/A) interferes with E2F1 nuclear export, K11- and K48-linked polyubiquitylation and degradation in differentiated keratinocytes. In contrast, replacement of S403 and T433 with phosphomimetic aspartic acid to generate a pseudophosphorylated E2F1 mutant protein (E2F1 ST/D) generates a protein that is regulated in a manner indistinguishable from that of wild type E2F1. Cdh1 is an activating cofactor that interacts with the anaphase-promoting complex/cyclosome (APC/C) ubiquitin E3 ligase, promoting proteasomal degradation of various substrates. We found that Cdh1 associates with E2F1 in keratinocytes. Inhibition or RNAi-mediated silencing of Cdh1 prevents E2F1 degradation in response to differentiation signals. Our results reveal novel regulatory mechanisms that jointly modulate post-translational modifications and downregulation of E2F1, which are necessary for proper epidermal keratinocyte differentiation.


All-atom and coarse-grained molecular dynamics simulations of a membrane protein stabilizing polymer.
Amphipathic polymers called amphipols (APols) have been developed as an alternative to detergents for stabilizing membrane proteins (MPs) in aqueous solutions. APols provide MPs with a particularly mild environment and, as a rule, keep them in a native functional state for longer periods than do detergents. Amphipol A8-35, a derivative of polyacrylate, is widely used and has been particularly well studied experimentally. In aqueous solutions, A8-35 molecules self-assemble into well-defined globular particles with a mass of ∼40 kDa and a R(g) of ∼2.4 nm. As a first step towards describing MP/A8-35 complexes by molecular dynamics (MD), we present three sets of simulations of the pure APol particle. First, we performed a series of all-atom MD (AAMD) simulations of the particle in solution, starting from an arbitrary initial configuration. Although AAMD simulations result in stable cohesive particles over a 45 ns simulation, the equilibration of the particle organization is limited. This motivated the use of coarse-grained MD (CGMD), allowing us to investigate processes on the microsecond time scale, including de novo particle assembly. We present a detailed description of the parametrization of the CGMD model from the AAMD simulations and a characterization of the resulting CGMD particles. Our third set of simulations utilizes reverse coarse-graining (rCG), through which we obtain all-atom coordinates from a CGMD simulation. This allows a higher-resolution characterization of a configuration determined by a long-timescale simulation. Excellent agreement is observed between MD models and experimental, small-angle neutron scattering data. The MD data provides new insight into the structure and dynamics of A8-35 particles, which is possibly relevant to the stabilizing effects of APols on MPs, as well as a starting point for modeling MP/A8-35 complexes.


Soybean trypsin inhibitor(s) reduce absorption of exogenous and increase loss of endogenous protein in miniature pigs.
It was the purpose of this study to define whether trypsin inhibitors impair protein digestibility via enhanced loss of exogenous or endogenous protein by quantifying those losses using the homoarginine technique, recently developed in this laboratory. Pigs fitted with permanent ileal T-cannulas were fed test meals containing homoarginine-labeled protein. The meals contained casein and increasing doses of trypsin inhibitors (Experiment 1) or alternatively either heat-treated or raw ground soybeans (Experiment 2). Following a casein meal (425 mmol nitrogen, no trypsin inhibitors), ileal protein was predominantly of endogenous rather than of exogenous origin (105 vs. 9 mmol nitrogen). Addition of isolated trypsin inhibitors (3000 mg) enhanced appearance of both endogenous and exogenous protein at the ileum (by 73 and 9 mmol nitrogen, respectively). Feeding raw instead of heat-treated soybeans in one single test meal caused a significant increase of endogenous protein from 217 +/- 42 to 263 +/- 47 mmol (mean +/- SEM) and of exogenous protein from 16 +/- 3 to 48 +/- 14 mmol. If fed continuously for 1 wk, a raw soybean diet caused endogenous protein loss to rise significantly from 221 +/- 26 to 432 +/- 85 mmol. We conclude that ingestion of food containing trypsin inhibitor affects nitrogen balance more by losses of amino acids of endogenous secreta than by losses of dietary amino acids.


Competition Reduces Response Times in Multiparty Conversation.
Natural conversations are characterized by short transition times between turns. This holds in particular for multi-party conversations. The short turn transitions in everyday conversations contrast sharply with the much longer speech onset latencies observed in laboratory studies where speakers respond to spoken utterances. There are many factors that facilitate speech production in conversational compared to laboratory settings. Here we highlight one of them, the impact of competition for turns. In multi-party conversations, speakers often compete for turns. In quantitative corpus analyses of multi-party conversation, the fastest response determines the recorded turn transition time. In contrast, in dyadic conversations such competition for turns is much less likely to arise, and in laboratory experiments with individual participants it does not arise at all. Therefore, all responses tend to be recorded. Thus, competition for turns may reduce the recorded mean turn transition times in multi-party conversations for a simple statistical reason: slow responses are not included in the means. We report two studies illustrating this point. We first report the results of simulations showing how much the response times in a laboratory experiment would be reduced if, for each trial, instead of recording all responses, only the fastest responses of several participants responding independently on the trial were recorded. We then present results from a quantitative corpus analysis comparing turn transition times in dyadic and triadic conversations. There was no significant group size effect in question-response transition times, where the present speaker often selects the next one, thus reducing competition between speakers. But, as predicted, triads showed shorter turn transition times than dyads for the remaining turn transitions, where competition for the floor was more likely to arise. Together, these data show that turn transition times in conversation should be interpreted in the context of group size, turn transition type, and social setting.


Effects of growth hormone and IGF-I on renal function in rats with normal and reduced renal mass.
To characterize actions of growth hormone (GH) and insulin-like growth factor ( (IGF-I) on renal function in rats with normal and reduced renal mass, we administered recombinant bovine growth hormone (bGH) or human IGF-I (hIGF-I) to normal rats or to rats that had undergone unilateral nephrectomy and two-thirds infarction of the contralateral kidney, and measured inulin and p-aminohippurate clearances over 10-17 days. Administration of either bGH (100-200 micrograms/day) or hIGF-I (200 micrograms/day) to rats with normal renal mass increased inulin and p-aminohippurate clearances compared with those measured in animals that received vehicle. Filtration fractions were not affected by either bGH or hIGF-I. Inulin clearance was decreased to approximately 17% of normal 1 day after reduction of renal mass in rats. Over the next 3 days insulin clearance increased significantly in rats with reduced renal mass that were administered vehicle. No further enhancement occurred during the next 7 days. Neither bGH nor hIGF-I affected inulin clearance in rats with reduced renal mass. We conclude that both GH and IGF-I enhance glomerular filtration rate when administered to rats with normal renal mass, but not when administered in the same quantities to rats in which renal functional mass is reduced. Glomerular filtration rate increases within 4 days of renal mass reduction independent of exogenous GH or IGF-I.