Quantification of glucuronidated and sulfated steroids in human urine by ultra-high pressure liquid chromatography quadrupole time-of-flight mass spectrometry.
The urinary steroid profile is constituted by anabolic androgenic steroids, including testosterone and its relatives, that are extensively metabolized into phase II sulfated or glucuronidated steroids. The use of liquid chromatography coupled to mass spectrometry (LC-MS) is an issue for the direct analysis of conjugated steroids, which can be used as urinary markers of exogenous steroid administration in doping analysis, without hydrolysis of the conjugated moiety. In this study, a sensitive and selective ultra high-pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometer (UHPLC-QTOF-MS) method was developed to quantify major urinary metabolites simultaneously after testosterone intake. The sample preparation of the urine (1 mL) was performed by solid-phase extraction on Oasis HLB sorbent using a 96-well plate format. The conjugated steroids were analyzed by UHPLC-QTOF-MS(E) with a single-gradient elution of 36 min (including re-equilibration time) in the negative electrospray ionization mode. MS(E) analysis involved parallel alternating acquisitions of both low- and high-collision energy functions. The method was validated and applied to samples collected from a clinical study performed with a group of healthy human volunteers who had taken testosterone, which were compared with samples from a placebo group. Quantitative results were also compared to GC-MS and LC-MS/MS measurements, and the correlations between data were found appropriate. The acquisition of full mass spectra over the entire mass range with QTOF mass analyzers gives promise of the opportunity to extend the steroid profile to a higher number of conjugated steroids.


The targeting of the proteasomal regulatory subunit S2 by adenovirus E1A causes inhibition of proteasomal activity and increased p53 expression.
Although adenovirus early region 1A (AdE1A) can modulate protein expression through its interaction with transcriptional regulators it can also influence the ability of the cell to degrade proteins by binding to components of the 26 S proteasome. We demonstrate here that AdE1A interacts with the S2 subunit of the 19 S regulatory complex in addition to the ATPase subunits S4 and S8 previously identified. S2 forms complexes with both the 13 and 12 S AdE1A proteins both in vivo and in vitro. Mutational analysis has shown direct binding through a short sequence toward the N terminus of conserved region 2 of AdE1A, which encompasses the LXCXE motif, involved in interaction with the pRb family of proteins. In vivo, additional contacts are made between AdE1A and proteasomal components, as well as within the proteasome, such that deletion of the N-terminal region of E1A as well as part of conserved region 2 is required to completely disrupt S2 binding. Mutation of AdE1A, which disrupts complex formation with S2, results in the loss of its ability to stabilize the p53 protein. Similarly down-regulation of S2 expression using small interfering RNAs leads to the inhibition of p53 degradation. These effects were observed in normally growing cells and those subjected to UV irradiation. Furthermore, AdE1A had no effect on the Mdm2-mediated ubiquitination of p53. We suggest therefore that interaction of AdE1A with S2, as well as with the ATPases S4 and S8, directly causes inhibition of proteasomal activity and consequent increase in the protein levels of p53.


Antibiotics or Tonsillectomy for Adult Recurrent Tonsillitis: Analyzing the Lesser of Two Evils.
To determine the best timing for surgical intervention for adults with recurrent tonsillitis (RT). A Markov model was constructed using variables and ranges based upon a literature review. A 1-way sensitivity analysis was performed to evaluate the number of yearly bouts at which each algorithm (antibiotics or tonsillectomy) would be favored. A Monte-Carlo probabilistic sensitivity analysis was calculated for gains and cost. Model outcomes were measured with quality-adjusted life-years (QALYs) and incremental cost-effectiveness ratios (ICER) for tonsillectomy versus repeat antibiotic treatment. Patients expected to sustain a single annual tonsillitis event will have a negative QALY of 0.02 if treated with surgery and those with 2 annual events will have a QALY gain from undergoing tonsillectomy of 0.01, 3 events = 0.03, 4 events = 0.05, 5 events = 0.07, 6 events = 0.09, 7 events = 0.1, and 8 events = 0.11. These gains became meaningful only after 2 years of recurrent bouts. The average cost of tonsillectomy was 3,238 USD, and the overall average cost of RT was 7,069 USD (an incremental cost of 3,831 USD). The ICER of tonsillectomy over antibiotic treatment for 1 QALY gain was 44,741 USD. Adult patients who sustain more than 3 annual bouts of tonsillitis over a period of at least 2 years will gain QALY after tonsillectomy. These gains increase proportionally to the number of yearly events and perennial episodes. The incremental costs of tonsillectomy fail to meet the NICE guidelines but are within other acceptable reference ranges. N/A Laryngoscope, 2023.


Quantum-dot-induced transparency in a nanoscale plasmonic resonator.
We investigate the near-field optical coupling between a single semiconductor nanocrystal (quantum dot) and a nanometer-scale plasmonic metal resonator using rigorous electrodynamic simulations. Our calculations show that the quantum dot produces a dip in both the extinction and scattering spectra of the surface-plasmon resonator, with a particularly strong change for the scattering spectrum. A phenomenological coupled-oscillator model is used to fit the calculation results and provide physical insight, revealing the roles of Fano interference and hybridization. The results indicate that it is possible to achieve nearly complete transparency as well as enter the strong-coupling regime for a single quantum dot in the near field of a metal nanostructure.


Endothelial and stem cell interactions on dielectrophoretically aligned fibrous silk fibroin-chitosan scaffolds.
Regenerative tissue engineering requires biomaterials that would mimic structure and composition of the extracellular matrix to facilitate cell infiltration, differentiation, and vascularization. Engineered scaffolds composed of natural biomaterials silk fibroin (SF) and chitosan (CS) blend were fabricated to achieve fibrillar nano-structures aligned in three-dimensions using the technique of dielectrophoresis. The effect of scaffold properties on adhesion and migration of human adipose-derived stem cells (hASC) and endothelial cells (HUVEC) was studied on SFCS (micro-structure, unaligned) and engineered SFCS (E-SFCS; nano-structure, aligned). E-SFCS constituted of a nano-featured fibrillar sheets, whereas SFCS sheets had a smooth morphology with unaligned micro-fibrillar extensions at the ends. Adhesion of hASC to either scaffolds occurred within 30 min and was higher than HUVEC adhesion. The percentage of moving cells and average speed was highest for hASC on SFCS scaffold as compared to hASC cocultured with HUVEC. HUVEC interactions with hASC appeared to slow the speed of hASC migration (in coculture) on both scaffolds. It is concluded that the guidance of cells for regenerative tissue engineering using SFCS scaffolds requires a fine balance between cell-cell interactions that affect the migration speed of cells and the surface characteristics that affects the overall adhesion and direction of migration.


Chart-stimulated Recall as a Learning Tool for Improving Radiology Residents' Reports.
Workplace-based assessments gauge the highest tier of clinical competence. Chart-stimulated recall (CSR) is a workplace-based assessment method that complements chart audit with an interview based on the residents' notes. It allows evaluation of the residents' knowledge and heuristics while providing opportunities for feedback and self-reflection. We evaluated the utility of CSR for improving the radiology residents' reporting skills. Residents in each year of training were randomly assigned to an intervention group (n = 12) or a control group (n = 13). Five pre-intervention and five post-intervention reports of each resident were independently evaluated by three blinded reviewers using a modified Bristol Radiology Report Assessment Tool. The study intervention comprised a CSR interview tailored to each individual resident's learning needs based on the pre-intervention assessment. The CSR process focused on the clinical relevance of the radiology reports. Student's t test (P < .05) was used to compare pre- and post-intervention scores of each group. A total of 125 pre-intervention and 125 post-intervention reports were evaluated (total 750 assessments). The Cronbach's alpha for the study tool was 0.865. A significant improvement was seen in the cumulative 19-item score (66% versus 73%, P < .001) and the global rating score (59% versus 72%, P < .001) of the intervention group after the CSR. The reports of the control group did not demonstrate any significant improvement. CSR is a feasible workplace-based assessment method for improving reporting skills of the radiology residents.


Transcatheter mitral valve replacement with Mi-thos system: First-in-human experience.
Transcatheter mitral valve replacement (TMVR) has become an alternative for high-risk patients with severe mitral regurgitation (MR). The aim of this study was to evaluate the safety and feasibility of the Mi-thos TMVR system (NewMed Medical) for high-risk patients with severe MR. This was a prospective, two-center, single-arm early feasibility study. Baseline characteristics, procedural data and 30-day follow-up outcomes were collected and analyzed. The primary endpoint was intraoperative success rate of device implantation. The second endpoints were all-cause mortality and major post-procedural complications. Echocardiographic data were evaluated by an independent core laboratory. Clinical events were adjudicated by a clinical events committee. Ten high-risk patients with severe MR were enrolled at two sites from August 2021 to November 2022. The median age was 70.5 years, and 60% of patients were female. The median Society of Thoracic Surgeons Predicted Risk of Mortality was 9.5%. The Mi-thos TMVR system was successfully implanted via transapical access in all patients. There was no pericedural mortality or major postpericedural complications during the 30-day follow-up. All implanted prosthetic valves had no or trace valvular or paravalvular MR, and the median mitral valve gradient at 30 days was 2.0 mmHg (IQR: 2.0-3.0 mmHg). There was one mild left ventricular outflow tract obstruction. The favorable short-term outcomes of the Mi-thos TMVR system demonstrated that it might be a feasible and safe therapeutic alternative for high-risk patients with severe MR. Nevertheless, further evaluation of the Mi-thos TMVR system is warranted.


Gender and occupation wise knowledge, Awareness and prevention of tuberculosis among people of district Muzaffarabad AJ & K.
To assess the awareness about the spread and control of tuberculosis as well as to investigate the gender and occupation wise differences among people regarding knowledge and attitude towards tuberculosis in the State of AJ & K. A cross-sectional descriptive study was conducted in district Muzaffarabad and a sample of 4000 respondents was selected by using stratified random sampling technique. The stratification was done with respect to gender and occupation. The occupation wise classification includes households, labors, and shop keepers, government employers, under graduate students of social and natural sciences, medical students and doctors. A close ended structured questionnaire was developed to collect the data and data were analyzed by using SPSS (Statistical Package for Social Sciences). Chi-Square test was used for association and Logistic Regression model was used to find out the most significant risk factors with gender. Majority of the males were more aware of tuberculosis than females regarding different aspects related to tuberculosis. The respondents from household, labors and shopkeepers have less awareness and knowledge than those who belong to other professions. The doctors and medical students have almost 100% awareness and knowledge of tuberculosis. It was examined that all the variables were associated with gender except threat, curable and transmissible. Only three variables mentioned above showed non- significant result, while all other variables were strongly associated with gender. Males were found more aware about TB than females. Moreover, the literate people were more conscious concerning the prevalence and threats of the disease.


Shock wave therapy for rotator cuff disease with or without calcification.
Shock wave therapy has seen widespread use since the 1990s to treat various musculoskeletal disorders including rotator cuff disease, but evidence of its efficacy remains equivocal. To determine the benefits and harms of shock wave therapy for rotator cuff disease, with or without calcification, and to establish its usefulness in the context of other available treatment options. We searched Ovid MEDLINE, Ovid Embase, CENTRAL, ClinicalTrials.gov and the WHO ICTRP up to November 2019, with no restrictions on language. We reviewed the reference lists of retrieved trials to identify potentially relevant trials. We included randomised controlled trials (RCTs) and controlled clinical trials (CCTs) that used quasi-randomised methods to allocate participants, investigating participants with rotator cuff disease with or without calcific deposits. We included trials of comparisons of extracorporeal or radial shock wave therapy versus any other intervention. Major outcomes were pain relief greater than 30%, mean pain score, function, patient-reported global assessment of treatment success, quality of life, number of participants experiencing adverse events and number of withdrawals due to adverse events. Two review authors independently selected studies for inclusion, extracted data and assessed the certainty of evidence using GRADE. The primary comparison was shock wave therapy compared to placebo. Thirty-two trials (2281 participants) met our inclusion criteria. Most trials (25) included participants with rotator cuff disease and calcific deposits, five trials included participants with rotator cuff disease and no calcific deposits, and two trials included a mixed population of participants with and without calcific deposits. Twelve trials compared shock wave therapy to placebo, 11 trials compared high-dose shock wave therapy (0.2 mJ/mm² to 0.4 mJ/mm² and above) to low-dose shock wave therapy. Single trials compared shock wave therapy to ultrasound-guided glucocorticoid needling, ultrasound-guided hyaluronic acid injection, transcutaneous electric nerve stimulation (TENS), no treatment or exercise; dual session shock wave therapy to single session therapy; and different delivery methods of shock wave therapy. Our main comparison was shock wave therapy versus placebo and results are reported for the 3 month follow up. All trials were susceptible to bias; including selection (74%), performance (62%), detection (62%), and selective reporting (45%) biases. No trial measured participant-reported pain relief of 30%. However, in one trial (74 participants), at 3 months follow up, 14/34 participants reported pain relief of 50% or greater with shock wave therapy compared with 15/40 with placebo (risk ratio (RR) 1.10, 95% confidence interval (CI) 0.62 to 1.94); low-quality evidence (downgraded for bias and imprecision). Mean pain (0 to 10 scale, higher scores indicate more pain) was 3.02 points in the placebo group and 0.78 points better (0.17 better to 1.4 better; clinically important change was 1.5 points) with shock wave therapy (9 trials, 608 participants), moderate-quality evidence (downgraded for bias). Mean function (scale 0 to 100, higher scores indicate better function) was 66 points with placebo and 7.9 points better (1.6 better to 14 better, clinically important difference 10 points) with shock wave therapy (9 trials, 612 participants), moderate-quality evidence (downgraded for bias). Participant-reported success was reported by 58/150 people in shock wave therapy group compared with 35/137 people in placebo group (RR 1.59, 95% CI 0.87 to 2.91; 6 trials, 287 participants), low-quality evidence (downgraded for bias and imprecision). None of the trials measured quality of life. Withdrawal rate or adverse event rates may not differ between extracorporeal shock wave therapy and placebo, but we are uncertain due to the small number of events. There were 11/34 withdrawals in the extracorporeal shock wave therapy group compared with 13/40 withdrawals in the placebo group (RR 0.75, 95% CI 0.43 to 1.31; 7 trials, 581 participants) low-quality evidence (downgraded for bias and imprecision); and 41/156 adverse events with extracorporeal shock wave therapy compared with 10/139 adverse events in the placebo group (RR 3.61, 95% CI 2.00 to 6.52; 5 trials, 295 participants) low-quality evidence (downgraded for bias and imprecision). Subgroup analyses indicated that there were no between-group differences in pain and function outcomes in participants who did or did not have calcific deposits in the rotator cuff. Based upon the currently available low- to moderate-certainty evidence, there were very few clinically important benefits of shock wave therapy, and uncertainty regarding its safety. Wide clinical diversity and varying treatment protocols means that we do not know whether or not some trials tested subtherapeutic doses, possibly underestimating any potential benefits. Further trials of extracorporeal shock wave therapy for rotator cuff disease should be based upon a strong rationale and consideration of whether or not they would alter the conclusions of this review. A standard dose and treatment protocol should be decided upon before further research is conducted. Development of a core set of outcomes for trials of rotator cuff disease and other shoulder disorders would also facilitate our ability to synthesise the evidence.


Computer vision-based helmet use registration for e-scooter riders - The impact of the mandatory helmet law in Copenhagen.
E-scooters are a new form of mobility used more frequently in urban environments worldwide. As there is evidence of an increased risk of head injuries, helmets are recommended and (less frequently) legislated. Denmark has enacted mandatory e-scooter helmet use legislation from January 1, 2022. So far, it is unclear how this newly implemented law influenced helmet use of e-scooter riders in Denmark immediately after its implementation. In this observational study, we register and compare e-scooter helmet use before the mandatory helmet use legislation (December 2021) and after (February 2022). As observational survey data collection in the field can be highly time-consuming, we conducted a video-based observation survey. We trained and applied a computer vision algorithm to automatically register e-scooter helmet use in the video data. The trained algorithm produces accurate helmet use data, which does not differ significantly from human-registered helmet use. In applying the algorithm to video data collected in December 2021 and February 2022, we register an overall e-scooter helmet use of 4.4% in n = 1054 riders. Splitting the observation between the time before and after the implementation of the helmet use law reveals a significant increase in helmet use from 1.80% to 5.56%. In this study, we successfully train and apply an object detection algorithm to register accurate helmet use data in videos collected in Copenhagen, Denmark. Using this algorithm, we find a significant impact of a new mandatory e-scooter helmet use law on e-scooter riders' helmet use behavior. Limitations of the study as well as future research needs, are discussed. Computer vision algorithms can be used for accurate e-scooter helmet assessments. Implementing a mandatory helmet use law can increase helmet use of e-scooters at specific observation sites.


Nandrolone decanoate inhibits gluconeogenesis and decreases fasting glucose in Wistar male rats.
The use of anabolic-androgenic steroids to improve physical performance or appearance has increased notably. The doses used are 10- to 100- fold higher than the therapeutic dose (TD), and this abuse can cause several side effects. Glucose metabolism is significantly affected by anabolic-androgenic steroid abuse, but studies about glycemic regulation during fasting are scarce. There are some evidences showing that testosterone can antagonize glucocorticoids action, which are crucial to glucose production during fasting. Thus, the aim of this study was to determine the impact of supraphysiological doses (SDs) of nandrolone decanoate (DECA) on rat glucose metabolism during fasting. Male Wistar rats were treated with i.m. injections of vehicle, a low TD (0.016 mg/100 g b.w.-TD group) or a high SD (1 mg/100 g b.w.-SD group) of DECA, once a week for 8 weeks. After 12 h fasting, we evaluated glucose and pyruvate tolerance tests, liver glycogen content, serum levels of gluconeogenic substrates, insulin and corticosterone, glucose uptake and hexokinase (HK) activity in skeletal muscle, and the adrenal catecholamine content. SD group had increased serum insulin levels and a blunted response to insulin regarding glucose uptake in skeletal muscle. Fasting serum glucose decreased significantly in SD group, as well as the pyruvate tolerance test and liver glycogen content. Moreover, serum levels of glycerol were increased in SD group. Our data indicate that SDs of DECA exert effects on different regulatory points of glucose metabolism, resulting in defective gluconeogenesis and decreased skeletal muscle glucose uptake in response to insulin.


Recording the parallelism of optically contacted Fabry-Perots.
A simple device has been constructed to record accurately the parallelism of optically contacted Fabry-Perots immediately after their manufacture. In this a Fabry-Perot fringe is tuned to be a maximum on-axis by varying the pressure of the gas in which the etalon is emersed. An insect-eye lens is then used to photograph the different fringe patterns from a large number of separate small areas of the Fabry-Perot simultaneously.


Assessment of some soil properties by spatial variability in saline and sodic soils in Arsanjan plain, Southern Iran.
Spatial patterns for several soil parameters such soil texture, Exchangeable Sodium Percentage (ESP), Electrical Conductivity (ECe), soil pH, Cation Exchange Capacity (CEC) were examined in saline and sodic soils in Arsanjan plain, Southern Iran, in order to identify their spatial distribution for implementation of a site-specific management. Soil samples were collected from 0-30, 30-60 and 60-90 cm soil depths at 85 sampling sites. Data were analyzed both statistically and geostatistically on the basis of the semivariogram. The spatial distribution model and spatial dependence level varied between soil parameters. Soil pH and ESP had the minimum and maximum variability at all depths, respectively. Soil properties indicated moderate to strong spatial dependence. ECe exhibited moderate spatial dependence at three depths; pH and ESP had a moderate spatial dependence at 0-30 cm and strong spatial dependence at 30-60 and 60-90 cm depths. Clay and CEC exhibited strong spatial dependence for the 0-30 cm and weak spatial dependence at 30-60 and 60-90 cm depths. Sand and silt had a non-spatial dependence at 0-30 cm and weak spatial dependency at 30-60 and 60-90 cm depths. The spatial variability in small distances of ECe, CEC, pH and ESP generally increased with depth. All geostatistical range values were greater than 1168 m. The results reported herein indicated that the strong spatial dependency of soil properties would lead to the extrinsic factors such as ground water level and drainage. It is important to know the spatial dependence of soil parameters, as management parameters with strong spatial dependence will be more readily managed and an accurate site-specific scheme for precision farming more easily developed.


Streptococcus entericus sp. nov., isolated from cattle intestine.
Biochemical, molecular chemical and molecular genetic studies were performed on an unknown gram-positive, catalase-negative, coccus-shaped organism isolated from the intestine of a cow affected with catarrhal enteritis. The organism was tentatively identified as a streptococcal species based on results of cellular morphological and biochemical tests. 16S rRNA gene sequencing studies confirmed its provisional identification as a member of the genus Streptococcus, but the organism did not correspond to any recognized species of this genus. The nearest phylogenetic relatives of the unknown coccus from a calf were Streptococcus acidominimus and Streptococcus suis. The unknown bacterium, however, was distinguished from these species and other animal streptococci by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on both phenotypic and phylogenetic findings, it is proposed that the unknown bacterium be classified as a novel species of the genus Streptococcus, Streptococcus entericus sp. nov. The type strain is CECT 5353T (= CCUG 44616T).


Neuroendocrine Abnormalities Following Traumatic Brain Injury: An Important Contributor to Neuropsychiatric Sequelae.
Neuropsychiatric symptoms following traumatic brain injury (TBI) are common and contribute negatively to TBI outcomes by reducing overall quality of life. The development of neurobehavioral sequelae, such as concentration deficits, depression, anxiety, fatigue, and loss of emotional well-being has historically been attributed to an ambiguous "post-concussive syndrome," considered secondary to frank structural injury and axonal damage. However, recent research suggests that neuroendocrine dysfunction, specifically hypopituitarism, plays an important role in the etiology of these symptoms. This post-head trauma hypopituitarism (PHTH) has been shown in the past two decades to be a clinically prevalent phenomenon, and given the parallels between neuropsychiatric symptoms associated with non-TBI-induced hypopituitarism and those following TBI, it is now acknowledged that PHTH is likely a substantial contributor to these impairments. The current paper seeks to provide an overview of hypothesized pathophysiological mechanisms underlying neuroendocrine abnormalities after TBI, and to emphasize the significance of this phenomenon in the development of the neurobehavioral problems frequently seen after head trauma.


Environmental exposure to PCBs and cancer incidence in eastern Slovakia.
In a population-based cross-sectional study, we compared serum levels of 15 polychlorinated biphenyl (PCB) congeners and three organochlorine pesticides in residents of two districts in eastern Slovakia, one with extensive environmental contamination from a former PCB production site (Michalovce) and the other matched on geography but with low (background) levels (Svidnik). The age-adjusted geometric means for sum of 15 measured PCB congeners were statistically significantly higher in subjects from the Michalovce district for both sexes: 3327.6 versus 1331.4 ng/g of lipid in males, 2751.8 versus 992.2 ng/g of lipid in females. Levels of DDE and DDT were also significantly higher, although the absolute differences were less pronounced. There was no substantial difference in HCB levels between the districts. We also conducted an ecologic study to compare cancer incidence occurring in these same two districts from 1985 through 1994. We calculated standardized incidence ratios (SIRs) and 95% confidence intervals for each district using indirect standardization and rates from eastern Slovakia. Among males from Michalovce (exposed), but not Svidnik (unexposed), there was an excess of cancer of the tongue (SIR=1.46; 1.06-1.96), stomach (SIR=1.15; 1.00-1.32), lung (SIR=1.14; 1.04-1.24), testis (SIR=1.40; 0.97-1.97), and kidney (SIR=1.23; 0.98-1.52), and lower than expected incidence of prostate cancer (SIR=0.83; 0.69-0.97); in contrast, there was an excess of peritoneal (SIR=3.05; 1.11-6.63) and laryngeal cancer (SIR=1.43; 0.99-1.98) in Svidnik not observed Michalovce. Among females from Michalovce, but not Svidnik, there was an excess of cancer of the lip (SIR=2.54; 1.53-3.96), stomach (SIR=1.22; 1.02-1.44), and lung (SIR=1.17; 0.94-1.45); in contrast, there was an excess of kidney (SIR=1.61; 1.03-2.40) and thyroid (SIR=1.97; 1.12-3.20) cancer in Svidnik not observed in Michalovce. Taken together, these results raise the possibility that high environmental exposure to organochlorines in the Michalovce district may be associated with higher rates of certain cancers, particularly stomach and lung cancer.


[Several methodologic approaches to determining the biological value of proteins].
By summarizing the results of their personal experimental research work and literature sources the authors come to the conclusion that a temporal determination of the free amino acids concentration in the portal vein blood of rats enables one to judge about the summary duration of the first two stages of the protein assimilation, namely about the rate of the protein digestion and that of the amino acids absorption. The aminogram of free amino acids in the blood of the portal vein shows a marked tendency toward convergence with the consumed protein aminogram.


Four siblings with achalasia, alacrimia and neurological abnormalities in a consanguineous family.
Four siblings with achalasia, alacrimia and other problems involving the autonomic nervous system involvements are reported. Achalasia and alacrimia were present in all of them. Their parents are first cousins and have four other healthy children. Electrophysiological tests showed that autonomic dysfunction has progressed with age. Blood cortisol levels were normal in all four affected children. Depending on those findings of our case and previous reports, we conclude that triple-A syndrome and achalasia, alacrimia with or without neurological abnormalities could be variable manifestations of the same autosomal recessive gene defect.


Semiconductor quantum dots for bioanalysis.
Semiconductor nanoparticles, or quantum dots (QDs), have unique photophysical properties, such as size-controlled fluorescence, have high fluorescence quantum yields, and stability against photobleaching. These properties enable the use of QDs as optical labels for the multiplexed analysis of immunocomplexes or DNA hybridization processes. Semiconductor QDs are also used to probe biocatalytic transformations. The time-dependent replication or telomerization of nucleic acids, the oxidation of phenol derivatives by tyrosinase, or the hydrolytic cleavage of peptides by proteases are probed by using fluorescence resonance energy transfer or photoinduced electron transfer. The photoexcitation of QD-biomolecule hybrids associated with electrodes enables the photoelectrochemical transduction of biorecognition events or biocatalytic transformations. Examples are the generation of photocurrents by duplex DNA assemblies bridging CdS NPs to electrodes, and by the formation of photocurrents as a result of biocatalyzed transformations. Semiconductor nanoparticles are also used as labels for the electrochemical detection of DNA or proteins: Semiconductor NPs functionalized with nucleic acids or proteins bind to biorecognition complexes, and the subsequent dissolution of the NPs allows the voltammetric detection of the related ions, and the tracing of the recognition events.


Anabolic steroids are fool's gold.
Since increases in muscle strength are proportional to increases in the cross-sectional diameter of the muscles being trained, the body must convert greater than normal amounts of amino acids available to it to increase size in athletes in training. When androgens became available in the 1930's they were used primarily to restore positive nitrogen balance in victims of starvation. Anabolic steroids, which were developed to avoid unwanted effects of androgens, were first given to weight lifters, but football players and weight throwers were soon using them. From 1965 to 1977, 25 clinical studies were published dealing with the administration of an anabolic-androgenic steroid to adult human males for evaluating changes in strength and, in 10 of these studies, in maximum oxygen consumption. In 12 of these studies, improvements were claimed from the use of these steroids; in the other 13 no improvements were observed. Other studies have shown that in healthy adult males these steroids reduce testosterone and gonadotrophin output, which reduces spermatogenesis. Alterations of normal liver function have been found in up to 80% of persons treated with C17-alkylated testosterone derivatives. Peliosis hepatitis, with liver failure and death, and fatal liver cancer have also been reported in adults so treated. Reliable methods for detecting anabolic steroids in the urine are now used in certain international competitions. Testing, announced bans, and disqualifications have not been effective in controlling the use of the drugs. The best hope for doing so lies in continuing education of athletes and their supervisors.


Principles and applications of flow injection analysis in biosensors.
In practical applications biosensors are often forced to operate under less than optimal conditions. Because of their construction, and the physical processes and chemical reactions involved in their operation, compromise conditions are frequently required to synchronize all events taking place. Therefore, and in order to implement functions such as periodic calibration, conditioning and possible regeneration of the biosensor, and, very importantly, to yield the freedom to select the optimum detection means, it is advantageous to use these devices in a flow-through mode, particularly by employing the flow injection (FI) approach. The capacity of FI, as offering itself as a complementary facility to augment the performance of biosensors, and in many cases as an attractive alternative, is demonstrated by reference to selected examples, comprising assays based on enzymatic procedures with optical and thermal detection procedures, and via description of a recently introduced technique for immunoassays, termed flow injection renewable surface immunoassays, which promises to entail powerful potentials and to yield compatible or better economy of operation than existing approaches.


Expression of glucagon receptors on T- and B- lymphoblasts: comparison with insulin receptors.
Activation of T- and B-lymphocytes by a variety of immunological stimuli has been reported to induce specific insulin receptors. The purpose of the present work was to determine whether glucagon receptors are also induced in activated cells. Studies of glucagon and insulin receptors were carried out using normal human mononuclear cells activated by phytohemagglutinin or T-cell growth factor (TCGF), as well as established B- and T-lymphoblastoid cell lines. With phytohemagglutinin, glucagon and insulin binding increased 15- and 36-fold, respectively, and peaked after 5 days in parallel with the rise in thymidine incorporation. Increased binding was associated with an increase in the number of receptors, most marked for insulin, though affinity for the insulin receptor was decreased. Normal human mononuclear cells cultured with TCGF showed an early modest rise in insulin binding due to increased receptor number, without a change in affinity, and a striking and progressive rise up to 50-fold in glucagon binding due to both increased receptor number and affinity. The differences in receptor response to these T-cell mitogens suggest that TCGF selects out a T-lymphoblast subset with very high glucagon receptors. B- and T-lymphoblastoid cells showed patterns of glucagon and insulin receptors that appear to be characteristic for each cell type. Glucagon binding was 7-fold higher (P less than 0.01), while inulin binding was 7-fold lower (P less than 0.01) in T- vs. B-lymphoblastoid cells. T-Cell lines had twice the number of glucagon receptors, whereas B-lines had 4-fold the number of insulin receptors, with much greater affinity for insulin compared with T-line insulin receptors. Induction of both insulin and glucagon receptors on activated lymphoblasts suggests that these receptors may play a significant role in cell function.


Extradural haematoma following temporomandibular joint arthrocentesis and lavage.
We describe a case of a 59-year-old woman who remained drowsy and developed a left hemiparesis following right temporomandibular joint (TMJ) arthrocentesis and lavage for TMJ dysfunction. CT demonstrated an extradural haematoma. This is the first reported case of an extradural haematoma following TMJ arthrocentesis.


The single biopsy approach is reliable for the measurement of muscle protein synthesis rates in vivo in older men.
We aimed to assess the reliability of the single biopsy approach for calculating muscle protein synthesis rates compared with the well described sequential muscle biopsy approach following a primed continuous infusion of L-[ring-(2)H(5)]phenylalanine and GC-MS analysis in older men. Two separate experimental infusion protocols, with differing stable isotope amino acid incorporation times, were employed consisting of n = 27 (experiment 1) or n = 9 (experiment 2). Specifically, mixed muscle protein FSR were calculated from baseline plasma protein enrichments and muscle protein enrichments obtained at 90 min or 50 min (1BX SHORT), 210 min or 170 min (1BX LONG), and between the muscle protein enrichments obtained at 90 and 210 min or 50 min and 170 min (2BX) of the infusion for experiments 1 and 2, respectively. In experiment 2, we also assessed the error that is introduced to the single muscle biopsy approach when nontracer naive subjects are recruited for participation in a primed continuous infusion of isotope-labeled amino acids. In experiment 1, applying the individual plasma protein enrichment values to the single muscle biopsy approach resulted in no differences in muscle protein FSR between the 1BX SHORT (0.031 ± 0.003%·h(-1)), 1BX LONG (0.032 ± 0.002%·h(-1)), or the 2BX approach (0.034 ± 0.002%·h(-1)). A significant correlation in muscle protein FSR was observed only between the 1BX LONG and 2BX approach (r = 0.8; P < 0.001). Similar results were observed in experiment 2. In addition, using the single biopsy approach in nontracer naïve state results in a muscle protein FSR that is negative for both the 1BX SHORT (-0.67 ± 0.051%·h(-1)) and 1BX LONG (-0.19 ± 0.051%·h(-1)) approaches. This is the first study to demonstrate that the single biopsy approach, coupled with the background enrichment of L-[ring-(2)H(5)]-phenylalanine of mixed plasma proteins, generates data that are similar to using the sequential muscle biopsy approach in the elderly population.


High-Order Areas and Auditory Cortex Both Represent the High-Level Event Structure of Music.
Recent fMRI studies of event segmentation have found that default mode regions represent high-level event structure during movie watching. In these regions, neural patterns are relatively stable during events and shift at event boundaries. Music, like narratives, contains hierarchical event structure (e.g., sections are composed of phrases). Here, we tested the hypothesis that brain activity patterns in default mode regions reflect the high-level event structure of music. We used fMRI to record brain activity from 25 participants (male and female) as they listened to a continuous playlist of 16 musical excerpts and additionally collected annotations for these excerpts by asking a separate group of participants to mark when meaningful changes occurred in each one. We then identified temporal boundaries between stable patterns of brain activity using a hidden Markov model and compared the location of the model boundaries to the location of the human annotations. We identified multiple brain regions with significant matches to the observer-identified boundaries, including auditory cortex, medial prefrontal cortex, parietal cortex, and angular gyrus. From these results, we conclude that both higher-order and sensory areas contain information relating to the high-level event structure of music. Moreover, the higher-order areas in this study overlap with areas found in previous studies of event perception in movies and audio narratives, including regions in the default mode network.


Optimization of partition-based Weighted Sum filters and their application to image denoising.
Partition-based Weighted Sum (P-WS) filtering is an effective method for processing nonstationary signals, especially those with regularly occurring structures, such as images. P-WS filters were originally formulated as Hard-partition Weighted Sum (HP-WS) filters and were successfully applied to image denoising. This formulation relied on intuitive arguments to generate the filter class. Here we present a statistical analysis that justifies the use of weighted sum filters after observation space partitioning. Unfortunately, the HP-WS filters are nondifferentiable and an analytical solution for their global optimization is therefore difficult to obtain. A two-stage suboptimal training procedure has been reported in the literature, but prior to this research no evaluation on the optimality of this approach has been reported. Here, a Genetic Algorithm (GA) HP-WS optimization procedure is developed that, in simulations, shows that the simpler two-stage training procedure yields near optimal results. Also developed in this paper are Soft-partition Weighted Sum (SP-WS) filters. The SP-WS filters utilize soft, or fuzzy, partitions that yield a differentiable filtering operation, enabling the development of gradient-based optimization procedures. Image denoising simulation results are presented comparing HP-WS and SP-WS filters, their optimization procedures, and wavelet-based image denoising. These results show that P-WS filters, in general, outperform traditional and wavelet-based image filters, and SP-WS filters utilizing soft partitioning not only allow for simple optimization, but also yields improved performance.


Endoplasmic reticulum stress induces epithelial-mesenchymal transition through autophagy via activation of c-Src kinase.
Endoplasmic reticulum (ER) stress has been implicated in inducing epithelial-mesenchymal transition (EMT). ER stress is also known to induce autophagy. However, it is unclear whether ER stress-induced autophagy contributes to EMT. We hypothesized that ER stress might induce EMT through autophagy via activation of c-Src kinase in tubular epithelial cells. All experiments were performed using HK-2 cells. Protein expression was measured by Western blot analysis. Immunofluorescence and small interfering RNA (siRNA) experiments were performed. Chemical ER stress inducers such as tunicamycin (TM, 0.2 μM) and thapsigargin (TG, 0.2 μM) induced EMT, as shown by upregulation of α-smooth muscle actin and downregulation of E-cadherin. ER stress inhibitors such as 4-PBA and salubrinal suppressed both TM- and TG-induced EMT. TM and TG also induced autophagy, as evidenced by upregulation of LC3-II and beclin-1, which were abolished by pretreatment with ER stress inhibitors. Transfection with siRNA targeting ER stress protein (IRE-1) blocked the TM- or TG-induced EMT and autophagy. Autophagy inhibitors such as 3-methyladenine and bafilomycin inhibited the TM- or TG-induced EMT. Transfection with siRNA targeting autophagy protein (beclin-1) also blocked the TM- or TG-induced EMT. Both TM and TG induced activation of c-Src kinase. Inhibitor of c-Src kinase (PP2) suppressed the TM- or TG-induced autophagy and EMT. ER stress by TM or TG induced EMT through autophagy via activation of c-Src kinase in tubular epithelial cells.


Altered nitrogenase MoFe proteins from Azotobacter vinelandii. Analysis of MoFe proteins having amino acid substitutions for the conserved cysteine residues within the beta-subunit.
The regions surrounding the three strictly conserved cysteine residues (positions 70, 95 and 153) in the beta-subunit of the Azotobacter vinelandii nitrogenase MoFe protein have been proposed to provide P-cluster environments [Dean, Setterquist, Brigle, Scott, Laird & Newton (1990) Mol. Microbiol. 4, 1505-1512]. In the present study, each of these cysteine residues was individually substituted by either serine or alanine by site-directed mutagenesis of the nifK gene, which encodes the MoFe protein beta-subunit. A mutant strain for which the codon for Cys-153 is removed was also isolated. Significant structural or functional roles are indicated for the cysteine residues at positions 70 and 95, where substitution by either serine or alanine eliminates diazotrophic growth of the resulting strains and abolishes or markedly decreases both MoFe-protein acetylene-reduction activity and the intensity of the whole-cell S = 3/2 e.p.r. signal. Changes introduced at position 153 have various effects on the functional properties of the enzyme. The strains produced either by deletion of the Cys-153 residue or its substitution by serine exhibit only a moderate decrease in diazotrophic growth and MoFe-protein activity and no loss of the whole-cell e.p.r.-signal intensity. In contrast, substitution by alanine eliminates diazotrophic growth and very markedly decreases both MoFe-protein activity and e.p.r.-signal intensity. These results are interpreted in terms of a metallocluster-driven protein rearrangement. After purification of the altered MoFe protein, in which serine replaces Cys-153, an investigation of its catalytic and spectroscopic properties confirms that neither the FeMo cofactor, i.e. the substrate-reduction site, nor the component-protein interaction site has been affected. Instead, these data indicate a disruption in electron transfer within the MoFe protein, which is consistent with a role for this residue (and region) at the P clusters.


Compulsive exercise: links, risks and challenges faced.
Compulsive exercise is a condition described since 1970s. It is characterized by a craving for physical training, resulting in uncontrollable excessive exercise behavior with harmful consequences, such as injuries and impaired social relations. It has not been accepted as a mental disorder in either International Classification of Diseases or Diagnostic and Statistical Manual of Mental Disorders. The aim of this literature review was to critically examine the research on links (comorbidity), risks (negative consequences), and challenges faced (problems in a treatment context). This review found that compulsive exercise is associated with eating disorder pathology, perfectionism, neuroticism, narcissism, and obsessive compulsive traits. The most prominent negative consequences were injuries, social impairment, and depression, but more research is needed to uncover the potential dysfunction resulting from compulsive exercise. As the condition is not recognized as a psychiatric disorder, studies on treatment interventions are sparse. Problems with compliance have been reported; therefore, motivational interviewing has been proposed as a treatment approach, in combination with cognitive behavioral therapy. This review summarizes and discusses findings on links/comorbidity, risks/negative consequences, and treatment challenges. We suggest that future studies should pay attention to both prevention and counseling in sports settings, where compulsive exercise appears, as the condition may be associated with harmful consequences.


Salivary alterations in type 2 (non-insulin-dependent) diabetes mellitus and hypertension.
The aim of this study was to determine whether saliva output and composition are altered in type 2 diabetes mellitus by comparison with a healthy, non-medicated control group, and also a group of hypertensives. From a community-dwelling cohort of Mexican American and European American subjects enrolled in the OH:SALSA oral aging study, we identified 233 subjects with type 2 diabetes mellitus, 227 with hypertension, and 240 healthy control subjects. We collected unstimulated whole (UW) and submandibular/ sublingual (US) saliva, as well as stimulated parotid (SP) and submandibular/ sublingual (SS) saliva. Flow rates were determined, yeast carriage was assayed in UW saliva, and SP and SS saliva samples were analyzed for protein composition. ELISA was used to determine concentrations of an array of specific protein components, with both antimicrobial and other activities. Both diabetic and hypertensive subjects had reduced output of both stimulated and unstimulated submandibular/sublingual saliva. 30% of the diabetic subjects had high oral yeast counts (> or =1000 cfu/mL) compared with 17% of the healthy subjects and 20% of the hypertensives. Significant increases in the concentrations of a number of the protein components were found in the diabetic subjects, specifically, SP lactoferrin, myeloperoxidase (MPO), and salivary peroxidase (SPO), as well as SS total protein, albumin, lactoferrin and secretory IgA. The pattern of decreased flow rates and increased protein concentrations were similar, but consistently greater in diabetics than hypertensives, suggesting that disease-specific mechanisms may be responsible. Diabetics may be more prone to oral dryness and infections than non-diabetics.


Basic features of biomedical assays.
All biomedical assays have certain aspects in common, the most striking of which is that they generally involve protein-binding. Ligand-ligator interactions and the resulting response form the basis of most assays. This paper reviews the advantages and limitations of different types of assays ranging from long-term bioassays to in vitro assays using cell-free components. Consideration of these aspects helps in the understanding of the different assays, selection of appropriate assays for different applications, and the better interpretation of assay results. Clinical chemists, hematologists, endocrinologists, immunologists, and scientists from other disciplines have each developed their own measurement methods and units. This paper seeks to identify those aspects and principles common to assays in all such disciplines, concentrating on protein-binding systems. Understanding what and how different assays measure may clarify why different assay methods often give different results.


The role of oscillations and synchrony in cortical networks and their putative relevance for the pathophysiology of schizophrenia.
Neural oscillations and their synchronization may represent a versatile signal to realize flexible communication within and between cortical areas. By now, there is extensive evidence to suggest that cognitive functions depending on coordination of distributed neural responses, such as perceptual grouping, attention-dependent stimulus selection, subsystem integration, working memory, and consciousness, are associated with synchronized oscillatory activity in the theta-, alpha-, beta-, and gamma-band, suggesting a functional mechanism of neural oscillations in cortical networks. In addition to their role in normal brain functioning, there is increasing evidence that altered oscillatory activity may be associated with certain neuropsychiatric disorders, such as schizophrenia, that involve dysfunctional cognition and behavior. In the following article, we aim to summarize the evidence on the role of neural oscillations during normal brain functioning and their relationship to cognitive processes. In the second part, we review research that has examined oscillatory activity during cognitive and behavioral tasks in schizophrenia. These studies suggest that schizophrenia involves abnormal oscillations and synchrony that are related to cognitive dysfunctions and some of the symptoms of the disorder. Perspectives for future research will be discussed in relationship to methodological issues, the utility of neural oscillations as a biomarker, and the neurodevelopmental hypothesis of schizophrenia.


Clinical trials carried out to assess non-parenteral routes for administration of Wistar RA 27/3 strain live attenuated rubella vaccine.
The problems encountered in obtaining acceptable seroconversion rates following intranasal and buccal administration of Wistar RA 27/3 strain live attenuated rubella vaccine are discussed with reference to vaccination technique, virus dose and other factors. Results from buccal administration of vaccine were not encouraging. However, intranasal vaccination may be acceptable if devices can be developed for the administration of vaccine which give reproducible sero-conversion rates equivalent to those obtained by subcutaneous vaccination.


Electrocardiographic findings associated with very low calorie dieting.
To determine the safety of very low calorie diets (VLCD) in regard to their effects on cardiac function. EKG changes were analyzed for 126 women on a VLCD of 3349 kJ/d (800 kcal/d). EKGs were done when the diet was begun, after 3 months of dieting, and at a 6 month follow up after being off the diet for 3 months. Subjects were solicited through advertisements and charged $1,000 for participating after being screened for age, weight, and health status. EKG QTc intervals, PR interval, QRS interval, ST-T wave changes, and heart rate. Over one-fourth (27.0%) of subjects had normal EKGs at all three time points studied. Sinus bradycardia was the most common abnormality, observed in 60 subjects (47.6%) on at least one of the three EKGs. Fifty-eight (46%) patients had EKGs with ST-T wave abnormalities observed on at least one of the EKGs. Eight subjects (6.4%) had prolonged QTc (more than one standard deviation beyond the average for women) intervals on at least one EKG. None of these eight persons had significant untoward medical consequences. A VLCD diet of 3349 kJ/d (800 kcal/d) for up to 3 months is not associated with significant electrocardiographic abnormalities or clinical cardiac complications, provided the patients have low cardiovascular risk at baseline.


Antibiotic Susceptibility Profiles of Lactic Acid Bacteria from the Human Vagina and Genetic Basis of Acquired Resistances.
Lactic acid bacteria can act as reservoirs of antibiotic resistance genes that can be ultimately transferred to pathogens. The present work reports on the minimum inhibitory concentration (MIC) of 16 antibiotics to 25 LAB isolates of five Lactobacillus and one Bifidobacterium species from the human vagina. Acquired resistances were detected to kanamycin, streptomycin, chloramphenicol, gentamicin, and ampicillin. A PCR analysis of lactobacilli failed to identify genetic determinants involved in any of these resistances. Surprisingly, a tet(W) gene was detected by PCR in two Bifidobacterium bifidum strains, although they proved to be tetracycline-susceptible. In agreement with the PCR results, no acquired genes were identified in the genome of any of the Lactobacillus spp. strains sequenced. A genome analysis of B. bifidum VA07-1AN showed an insertion of two guanines in the middle of tet(W) interrupting the open reading frame. By growing the strain in the presence of tetracycline, stable tetracycline-resistant variants were obtained. An amino acid substitution in the ribosomal protein S12 (K43R) was further identified as the most likely cause of VA07-1AN being streptomycin resistance. The results of this work expand our knowledge of the resistance profiles of vaginal LAB and provide evidence for the genetic basis of some acquired resistances.


Uniform 2 nm gold nanoparticles supported on iron oxides as active catalysts for CO oxidation reaction: structure-activity relationship.
Uniform Au nanoparticles (∼2 nm) with narrow size-distribution (standard deviation: 0.5-0.6 nm) supported on both hydroxylated (Fe_OH) and dehydrated iron oxide (Fe_O) have been prepared by either deposition-precipitation (DP) or colloidal-deposition (CD) methods. Different structural and textural characterizations were applied to the dried, calcined and used gold-iron oxide samples. Transmission electron microscopy (TEM) and high-resolution TEM (HRTEM) showed high homogeneity in the supported Au nanoparticles. The ex situ and in situ X-ray absorption fine structure (XAFS) characterization monitored the electronic and short-range local structure of active gold species. The synchrotron-based in situ X-ray diffraction (XRD), together with the corresponding temperature-programmed reduction by hydrogen (H2-TPR), indicated a structural evolution of the iron-oxide supports, correlating to their reducibility. An inverse order of catalytic activity between DP (Au/Fe_OH < Au/Fe_O) and CD (Au/Fe_OH > Au/Fe_O) was observed. Effective gold-support interaction results in a high activity for gold nanoparticles, locally generated by the sintering of dispersed Au atoms on the oxide support in the DP synthesis, while a hydroxylated surface favors the reactivity of externally introduced Au nanoparticles on Fe_OH support for the CD approach. This work reveals why differences in the synthetic protocol translate to differences in the catalytic performance of Au/FeOx catalysts with very similar structural characteristics in CO oxidation.


Focus issue: TGF-β and the mesenchymal transition in physiology and disease.
The ability of cells to alter their phenotypic and morphological characteristics, known as cellular plasticity, is critical in normal embryonic development and adult tissue repair and contributes to the progression of diseases like organ fibrosis and cancer. Signaling mediated by the transforming growth factor-β (TGF-β) superfamily of extracellular protein ligands is one pathway that is integral to these processes. Content published in Science Signaling highlights the roles and complexities of TGF-β signaling in physiology and disease.


Changes of kinematic parameters of lower extremities with gait speed: a 3D motion analysis study.
[Purpose] The purpose of this study was to investigate the changes in hip, knee and ankle kinematic variables of the lower extremities at different gait speeds. [Subjects and Methods] Forty healthy subjects who had no previous history of neurological, musculo-skeletal or other medical conditions that could affect gait were recruited. The subjects were asked to walk 10 m down a walkway at three different gait speeds: normal gait speed, and self-selected fast, and slow speeds. The experimental order was randomly chosen across these gaits. The hip, knee and ankle kinematic data were evaluated using a VICON 3D motion analysis system and force plates. [Results] The flexion peak and external rotation peak of the knee joint significantly increased with the increase of gait speed. The plantarflexion peaks of the ankle joint significantly increased with increase of gait speed. However, none of the kinematic data of the hip joint were significantly dependent on increase of gait speed. [Conclusion] The relationship of the knee and ankle joint can be described as coupling motion which is dependent on gait speed. Our present findings suggest that coupling motion of the knee joint and plantarflexion of the ankle joint significantly increase with increase of gait speed. These results will provide important insight into gait mechanisms for the evaluation of pathological populations.


Keloid microRNA expression analysis and the influence of miR-199a-5p on the proliferation of keloid fibroblasts.
The purpose of this study was to identify microRNAs (miRNAs) involved in keloid formation and determine their influence on the proliferation of keloid fibroblasts (KFs). Eight specimens each of resected keloid tissue and normal skin tissue were collected. miRNAs that are differentially expressed in keloid tissue and normal skin were detected using an miRNA microarray and verified by quantitative real-time polymerase chain reaction (RT-PCR). Seventeen differentially expressed miRNAs, including miR-199a-5p, were identified by microarray hybridization. qRT-PCR analysis confirmed the decrease in miR-199a-5p expression in keloid vs normal tissue that was detected by the microarray analysis. Mimics of differentially expressed miRNAs were then transfected into a KF cell line, and the effect of miRNA overexpression on the proliferation of KFs was assayed using the EdU assay. Compared with mock-transfected cells, KFs transfected with a miR-199a-5p mimic showed significantly lower cell proliferation and an altered cell cycle, with cells having significantly longer S and G2/M phases. The significantly lower expression of miRNA-199a-5p in keloids likely influences the cell cycle of KFs and restrains their proliferation, suggesting that miR-199a-5p probably plays a role in the regulation of KF proliferation.


cDNA cloning and molecular analysis of papaya small GTP-binding protein, pgp1.
In the course of papaya EST collection, one clone (pRA4-3) encoding partial sequence of papaya small GTP-binding protein gene, pgp1, was obtained. Based on the sequence information of pRA4-3, the entire coding region of pgp1 was cloned using the 3'RACE PCR technique. ORF of pgp1 is 636bp long and deduced molecular weight of the protein is 23,311. Phylogenetic analysis showed that PGP1 belongs to YPT/RAB group of the small GTP-binding protein and is a homologue of RAB2. Southern analysis showed that there are several pgp1-related genes in papaya genome. Northern analysis showed that pgp1 was expressed equally in stems of seedlings that were grown under light and dark conditions. This result shows that PGP1 is not involved in the phytochrome-mediated signal transduction as an auxin signal transducer in stems of papaya seedlings.


Generation of c-MycERTAM-transduced human late-adherent olfactory mucosa cells for potential regenerative applications.
Human olfactory mucosa cells (hOMCs) have been transplanted to the damaged spinal cord both pre-clinically and clinically. To date mainly autologous cells have been tested. However, inter-patient variability in cell recovery and quality, and the fact that the neuroprotective olfactory ensheathing cell (OEC) subset is difficult to isolate, means an allogeneic hOMC therapy would be an attractive "off-the-shelf" alternative. The aim of this study was to generate a candidate cell line from late-adherent hOMCs, thought to contain the OEC subset. Primary late-adherent hOMCs were transduced with a c-MycERTAM gene that enables cell proliferation in the presence of 4-hydroxytamoxifen (4-OHT). Two c-MycERTAM-derived polyclonal populations, PA5 and PA7, were generated and expanded. PA5 cells had a normal human karyotype (46, XY) and exhibited faster growth kinetics than PA7, and were therefore selected for further characterisation. PA5 hOMCs express glial markers (p75NTR, S100ß, GFAP and oligodendrocyte marker O4), neuronal markers (nestin and ß-III-tubulin) and fibroblast-associated markers (CD90/Thy1 and fibronectin). Co-culture of PA5 cells with a neuronal cell line (NG108-15) and with primary dorsal root ganglion (DRG) neurons resulted in significant neurite outgrowth after 5 days. Therefore, c-MycERTAM-derived PA5 hOMCs have potential as a regenerative therapy for neural cells.


Hosts mobility and spatial spread of Rickettsia rickettsii.
There are a huge number of pathogens with multi-component transmission cycles, involving amplifier hosts, vectors or complex pathogen life cycles. These complex systems present challenges in terms of modeling and policy development. A lethal tick-borne infectious disease, the Brazilian Spotted Fever (BSF), is a relevant example of that. The current increase of human cases of BSF has been associated with the presence and expansion of the capybara Hydrochoerus hydrochaeris, amplifier host for the agent Rickettsia rickettsii and primary host for the tick vector Amblyomma sculptum. We introduce a stochastic dynamical model that captures the spatial distribution of capybaras and ticks to gain a better understanding of the spatial spread of the R. rickettsii and potentially predict future epidemic outcomes. We implemented a reaction-diffusion process in which individuals were divided into classes denoting their state with respect to the disease. The model considered bidirectional movements between base and destination locations limited by the carrying capacity of the environment. We performed systematic stochastic simulations and numerical analysis of the model and investigate the impact of potential interventions to mitigate the spatial spread of the disease. The mobility of capybaras and their attached ticks was significantly influenced by the birth rate of capybaras and therefore, disease propagation velocity was higher in places with higher carrying capacity. Some geographical barriers, generated for example by riparian reforesting, can impede the spatial spread of BSF. The results of this work will allow the formulation of public actions focused on the prevention of BSF human cases.


A tandem GC box motif is necessary for lipopolysaccharide-induced transcription of the type II TNF receptor gene.
LPS induces the expression of the gene encoding the type II TNF-alpha receptor in mononuclear phagocytes. To elucidate the nuclear signaling mechanisms involved in this response, a 772-bp fragment of the TNFRII gene promoter was analyzed by deletion and site-specific mutagenesis following transient transfection in the macrophage-like cell line RAW264.7. A region located between -100 and -75 relative to the transcription start site was found to be essential for LPS sensitivity. This contained a GC-rich region composed of two tandemly arrayed Sp-1 sites. While mutation of this region confirmed that it was essential for LPS sensitivity, the sequence was not able to confer LPS sensitivity upon a heterologous promoter. Internal deletion and site-specific mutagenesis of the 100-bp fragment identified regions immediately flanking an initiator region (Inr) site that were also necessary for sensitivity to LPS. Electrophoretic mobility shift assays demonstrated that the GC box bound Sp-1 and Sp-3, although the level of binding activity did not vary with LPS stimulation. An oligonucleotide probe containing nucleotide positions -45 to -18 also bound several protein complexes, but these were not enhanced by LPS. These findings indicate that the tandem GC box is necessary, but not sufficient, for LPS-mediated transcription of the TNFRII gene. A second apparently novel motif, located within 15 nucleotides of the Inr, is also necessary.


Well-differentiated serous neoplasms of the ovary.
Serous neoplasms of the ovary, which constitute the largest subgroup of the surface epithelial tumors, cluster into three distinctly clinicopathologic groups: benign neoplasms, which are architecturally noncomplex, confined to the ovary, and composed of cytologically bland cells; carcinomatous neoplasms, which have spread beyond the ovary and are cytologically malignant; and an intermediate group, which raises serious problems in taxonomy, differential diagnosis, and prognosis. This chapter focuses on differential diagnosis, emphasizing the authors' experience as well as reports from other investigators.


The safety profile and antiviral activity of the combination of stavudine, didanosine, and nelfinavir in patients with HIV infection.
We assessed the safety profile, tolerability, and antiviral effect of 12 weeks of triple combination therapy with stavudine (d4T), didanosine (ddI), and nelfinavir in patients who had not previously received therapy with d4T, ddI, or a protease inhibitor. We also assessed the effect of the buffered tablet formulation of ddI on the pharmacokinetics of nelfinavir. The study had a single-arm, open-label design and enrolled patients aged > or =18 years who had HIV infection and > or =10,000 plasma HIV RNA copies/mL. Patients received the full recommended doses of oral d4T, ddI, and nelfinavir. Efficacy was assessed in terms of change from baseline in plasma HIV RNA and CD4+ cell counts, as well as in terms of the proportion of patients achieving HIV RNA levels <400 copies/mL. The first 10 patients enrolled in the study were included in a substudy to determine the effects of the buffered tablet formulation of ddI on the pharmacokinetic profile of nelfinavir. A dose of ddI was given 1 hour before nelfinavir, after which the maximum plasma concentration (Cmax), time to Cmax (Tmax), and area under the concentration-time curve (AUC) of nelfinavir were determined. A total of 22 patients entered the trial, of whom 1 (5%) had AIDS, 12 (55%) had symptomatic HIV infection, and 9 (41%) were asymptomatic. The median baseline CD4+ cell count was 315 cells/microL (range, 70-709 cells/microL), and the median plasma viral load was 4.8 log10 copies/mL (range, 4.0-5.6 log10 copies/mL). ddI had no clinically significant effects on the plasma pharmacokinetics of nelfinavir. At the end of 12 weeks of treatment, the mean (+/- SE) decrease in plasma viral load was 1.36+/-0.24 log10 copies/mL, and 8 of 16 patients (50%) achieved plasma HIV RNA levels <400 copies/mL; the mean (+/- SE) increase in CD4+ cell count was 111.4+/-31.7 cells/microL. Patients who were judged to be compliant with antiretroviral therapy (ie, who missed <7 days of all 3 study drugs during 12 weeks of treatment) experienced mean decreases in viral load exceeding 2.0 log10 copies/mL, and 6 of 7 patients achieved HIV RNA levels <400 copies/mL after 12 weeks of therapy. Although 95% of patients reported an adverse event of grade 1 or higher, only 1 patient experienced a grade 3 or 4 adverse event (maculopapular rash) related to nelfinavir. As reflected in the Cmax, Tmax, and AUC, administration of ddI 1 hour before nelfinavir did not influence the pharmacokinetic profile of the protease inhibitor. Triple drug therapy with d4T, ddI, and nelfinavir was well tolerated and associated with few clinically significant toxicities. This treatment resulted in substantial reductions in viral load and improvements in CD4+ cell count over 12 weeks.


Quasi-elastic Neutron Scattering Reveals Ligand-Induced Protein Dynamics of a G-Protein-Coupled Receptor.
Light activation of the visual G-protein-coupled receptor (GPCR) rhodopsin leads to significant structural fluctuations of the protein embedded within the membrane yielding the activation of cognate G-protein (transducin), which initiates biological signaling. Here, we report a quasi-elastic neutron scattering study of the activation of rhodopsin as a GPCR prototype. Our results reveal a broadly distributed relaxation of hydrogen atom dynamics of rhodopsin on a picosecond-nanosecond time scale, crucial for protein function, as only observed for globular proteins previously. Interestingly, the results suggest significant differences in the intrinsic protein dynamics of the dark-state rhodopsin versus the ligand-free apoprotein, opsin. These differences can be attributed to the influence of the covalently bound retinal ligand. Furthermore, an idea of the generic free-energy landscape is used to explain the GPCR dynamics of ligand-binding and ligand-free protein conformations, which can be further applied to other GPCR systems.


Helicase-primase inhibitor pritelivir for HSV-2 infection.
Pritelivir, an inhibitor of the viral helicase-primase complex, exhibits antiviral activity in vitro and in animal models of herpes simplex virus (HSV) infection. We tested the efficacy and safety of pritelivir in otherwise healthy persons with genital HSV-2 infection. We randomly assigned 156 HSV-2-positive persons with a history of genital herpes to receive one of four doses of oral pritelivir (5, 25, or 75 mg daily, or 400 mg weekly) or placebo for 28 days. Participants obtained daily swabs from the genital area for HSV-2 testing, which was performed with a polymerase-chain-reaction assay. Participants also maintained a diary of genital signs and symptoms. The primary end point was the rate of genital HSV shedding. HSV shedding among placebo recipients was detected on 16.6% of days; shedding among pritelivir recipients was detected on 18.2% of days among those receiving 5 mg daily, 9.3% of days among those receiving 25 mg daily, 2.1% of days among those receiving 75 mg daily, and 5.3% of days among those receiving 400 mg weekly. The relative risk of viral shedding with pritelivir, as compared with placebo, was 1.11 (95% confidence interval [CI], 0.65 to 1.87) with the 5-mg daily dose, 0.57 (95% CI, 0.31 to 1.03) with the 25-mg daily dose, 0.13 (95% CI, 0.04 to 0.38) with the 75-mg daily dose, and 0.32 (95% CI, 0.17 to 0.59) with the 400-mg weekly dose. The percentage of days with genital lesions was also significantly reduced, from 9.0% in the placebo group to 1.2% in both the group receiving 75 mg of pritelivir daily (relative risk, 0.13; 95% CI, 0.02 to 0.70) and the group receiving 400 mg weekly (relative risk, 0.13; 95% CI, 0.03 to 0.52). The rate of adverse events was similar in all groups. Pritelivir reduced the rates of genital HSV shedding and days with lesions in a dose-dependent manner in otherwise healthy men and women with genital herpes. (Funded by AiCuris; ClinicalTrials.gov number, NCT01047540.).


Population-specificity of heat stress gene induction in northern and southern eelgrass Zostera marina populations under simulated global warming.
Summer heat waves have already resulted in mortality of coastal communities, including ecologically important seagrass meadows. Gene expression studies from controlled experiments can provide important insight as to how species/genotypes react to extreme events that will increase under global warming. In a common stress garden, we exposed three populations of eelgrass, Zostera marina, to extreme sea surface temperatures, simulating the 2003-European heat wave. Populations came from locations widely differing in their thermal regime, two northern European locations [Ebeltoft (Kattegat), Doverodde (Limfjord, Baltic Sea)], and one southern population from Gabicce Mare (Adriatic Sea), allowing to test for population specificity in the response to a realistic heat stress event. Eelgrass survival and growth as well as the expression of 12 stress associated candidate genes were assessed during and after the heat wave. Contrary to expectations, all populations suffered equally from 3 weeks of heat stress in terms of shoot loss. In contrast, populations markedly differed in multivariate measures of gene expression. While the gene expression profiles converged to pre-stress values directly after the heat wave, stress correlated genes were upregulated again 4 weeks later, in line with the observed delay in shoot loss. Target genes had to be selected based on functional knowledge in terrestrial plants, nevertheless, 10/12 genes were induced relative to the control treatment at least once during the heat wave in the fully marine plant Z. marina. This study underlines the importance of realistic stress and recovery scenarios in studying the impact of predicted climate change.


Safety and feasibility of destination care for bariatric surgery: a single institution retrospective study.
Increasing emphasis on value-based healthcare has prompted both employers and healthcare organizations to develop innovative strategies to supply high quality care to patients. One such strategy is through the bundled care payment model (BCPM). Through this model, our institution partnered with employers from across the country to provide quality care for their members. Patients traveling greater than 2 h driving time from the bariatric center were considered "destination" patients. To properly care for our destination patients, our institution created a "destination bariatric program." We sought to investigate comparative outcomes for the first 100 patients who completed the program. We hypothesized that there would be no difference in patient outcomes or complications between destination and local patient groups undergoing sleeve gastrectomy (SG) or Roux-en-Y gastric bypass (RYGB). A retrospective cohort analysis of patients undergoing bariatric surgery at a MBSAQIP-accredited bariatric surgery center between May 2019 and October 2021 was conducted. Patients were divided into destination or local patient groups based on participation in the established destination surgery program. Patient demographics, perioperative clinical outcomes, and complications were compared and statistically analyzed using two-sample t-tests, Chi-square tests, Fisher's exact tests, and univariate logistic regressions. This study identified 296 patients, which consisted of destination (n = 110) and local (n = 186) patient cohorts. Patients in the destination group had higher rates of diabetes mellitus (29.1% vs 24.2%, p = 0.029), but otherwise cohorts had similar basic demographics and comorbidities. Outcomes revealed no statistically significant associations between patient cohort (destination versus local) and ED admission (p = 0.305), hospital readmission (p = 0.893), surgical reintervention (p = 0.974), endoscopic-reintervention (p = 0.714), and patient complications in the postoperative period (30 days). Participation in destination care programs for bariatric surgery was found to be both safe and feasible. These destination programs represent an opportunity to provide a broader patient population access to complex surgical care.


Oral pyogenic granuloma: Various concepts of etiopathogenesis.
Pyogenic granuloma or granuloma pyogenicum is a well-known oral lesion. The name pyogenic granuloma is a misnomer since the condition is not associated with pus and does not represent a granuloma histologically. Pyogenic granuloma of the oral cavity is known to involve the gingiva commonly. Extragingivally, it can occur on the lips, tongue, buccal mucosa, palate, and the like. A history of trauma is common in such sites. The etiology of the lesion is not known, though it was originally believed to be a botryomycotic infection. It is theorized that pyogenic granuloma possibly originates as a response of tissues to minor trauma and/or chronic irritation, thus opening a pathway for invasion of nonspecific microorganisms, although microorganisms are seldom demonstrated within the lesion. Pathogenesis of pyogenic granuloma is still debatable. Medline and PubMed databases were searched under the following key terms: Pathogenesis of oral pyogenic granuloma, pyogenic granuloma, and oral pyogenic granuloma. This search was limited to articles on human/animal studies which were published in English language. After reviewing the searched articles, the relevant articles were selected for the present review. Through this article, we have tried to summarize and present all the concepts of pathogenesis related to this most common and most mysterious oral lesion.


Elderberries: a source of ribosome-inactivating proteins with lectin activity.
Sambucus (Adoxaceae) species have been used for both food and medicine purposes. Among these, Sambucus nigra L. (black elder), Sambucus ebulus L. (dwarf elder), and Sambucus sieboldiana L. are the most relevant species studied. Their use has been somewhat restricted due to the presence of bioactive proteins or/and low molecular weight compounds whose ingestion could trigger deleterious effects. Over the last few years, the chemical and pharmacological characteristics of Sambucus species have been investigated. Among the proteins present in Sambucus species both type 1, and type 2 ribosome-inactivating proteins (RIPs), and hololectins have been reported. The biological role played by these proteins remains unknown, although they are conjectured to be involved in defending plants against insect predators and viruses. These proteins might have an important impact on the nutritional characteristics and food safety of elderberries. Type 2 RIPs are able to interact with gut cells of insects and mammals triggering a number of specific and mostly unknown cell signals in the gut mucosa that could significantly affect animal physiology. In this paper, we describe all known RIPs that have been isolated to date from Sambucus species, and comment on their antiviral and entomotoxic effects, as well as their potential uses.


Thermal sensitivity across forest vertical profiles: patterns, mechanisms, and ecological implications.
Rising temperatures are influencing forests on many scales, with potentially strong variation vertically across forest strata. Using published research and new analyses, we evaluate how microclimate and leaf temperatures, traits, and gas exchange vary vertically in forests, shaping tree, and ecosystem ecology. In closed-canopy forests, upper canopy leaves are exposed to the highest solar radiation and evaporative demand, which can elevate leaf temperature (Tleaf ), particularly when transpirational cooling is curtailed by limited stomatal conductance. However, foliar traits also vary across height or light gradients, partially mitigating and protecting against the elevation of upper canopy Tleaf . Leaf metabolism generally increases with height across the vertical gradient, yet differences in thermal sensitivity across the gradient appear modest. Scaling from leaves to trees, canopy trees have higher absolute metabolic capacity and growth, yet are more vulnerable to drought and damaging Tleaf than their smaller counterparts, particularly under climate change. By contrast, understory trees experience fewer extreme high Tleaf 's but have fewer cooling mechanisms and thus may be strongly impacted by warming under some conditions, particularly when exposed to a harsher microenvironment through canopy disturbance. As the climate changes, integrating the patterns and mechanisms reviewed here into models will be critical to forecasting forest-climate feedback.


Biogeographic Ancestry in the African Descent and Glaucoma Evaluation Study (ADAGES): Association With Corneal and Optic Nerve Structure.
We determined if quantitative measurements of biogeographic ancestry (BGA) correlate with variations in optic disc area, corneal thickness (CCT), and retinal nerve fiber layer (RNFL) thickness. Data were obtained from 656 participants in the African Descent and Glaucoma Evaluation Study (ADAGES) cohort who consented to BGA testing. Data for CCT, optic disc area, and RNFL thickness were obtained from subjects in the ADAGES study who also had participated in the current substudy. A total of 31 ancestry informative markers (AIMs) with large allele frequencies differences between populations was used to calculate admixture proportion (implemented in STRUCTURE). Correlations with BGA adjusted for diagnosis, age, and sex for CCT and optic disc area using the whole group and RNFL thickness adjusted for age and sex for the normal study participants were determined. The mean percentage of African admixture was 79.6% in the self-described African Descent (AD) group and 3.5% in the European Descent (ED) group. Percent African ancestry was significantly correlated with CCT (ρ = -0.27, P < 0.0001) and disc area (ρ = 0.15, P < 0.0001), but only marginally associated with RNFL thickness (ρ = 0.20, P = 0.092) in adjusted models. The BGA correlates with variation in ocular features that significantly differ across racial groups and that have been associated with the development of glaucoma. While BGA can provide an objective measurement of the biologic component of self-described race for ocular research, for most nongenetic epidemiologic studies, self-described race may adequately describe the associations with these ocular characteristics. (ClinicalTrials.gov number, NCT00221923.).


[Therapeutic plasma exchange in severe tropical malaria].
Cases of malaria are being seen ever more frequently in the Federal Republic of Germany, presenting both diagnostic and therapeutic problems. Three recent cases of severe malignant falciparum malaria are presented, one ending fatally. In the other two, plasma exchange combined with haemodialysis led to a cure.


Evidence of subtle gray-matter pathologic changes in healthy elderly individuals with nonspecific white-matter hyperintensities.
To investigate whether additional "occult" tissue changes can be detected in the normal-appearing white matter and gray matter of otherwise normal elderly individuals with nonspecific white-matter hyperintensities on conventional magnetic resonance images of the brain. Conventional and magnetization transfer magnetic resonance images were obtained from 12 otherwise normal elderly subjects with white-matter hyperintensities and 11 age- and sex-matched normal individuals. After automatic tissue segmentation, image coregistration, and masking of T2-visible lesions, we obtained magnetization transfer ratio histograms of the normal-appearing white matter and gray matter. For each histogram, the average magnetization transfer ratio, the peak height, and the peak position were measured. We also calculated the percentages of gray-matter and white-matter volumes normalized over the total volume of the intracranial content and the total normalized brain volumes. Average magnetization transfer ratio (P =.03) and mean peak position (P =.01) of the gray-matter histograms from elderly individuals with white-matter hyperintensities were significantly lower than the corresponding quantities from those without white-matter hyperintensities. The normalized percentages of gray and white matter and normalized brain volume did not differ between the 2 groups. The average gray-matter magnetization transfer ratio was correlated with the average lesion magnetization transfer ratio (r = 0.68; P<.01). This study shows that brain abnormalities in otherwise normal elderly subjects with nonspecific white-matter hyperintensities extend beyond the macroscopic white-matter lesions visualized on conventional magnetic resonance images.


Oxidation as a facile strategy to reduce the surface charge and toxicity of polyethyleneimine gene carriers.
Polyethyleneimine (PEI) is widely regarded as one of the most efficient non-viral transfection agents commercially available. However, a key concern is its pronounced cytotoxicity, ascribed mainly to its high amine content and cationic charge density. Significant past efforts to mitigate its toxicity usually involved lengthy synthetic procedures. We now propose a simple strategy using hydrogen peroxide (H2O2) to oxidize the amine groups. PEI/DNA complexes were first formed before some amine groups were removed with H2O2. This reduced surface charge while the remaining cationic charges still allowed for efficient transfection. The DNA was not damaged and remained bound after oxidation. Furthermore, H2O2 was quantitatively removed with sodium pyruvate prior to cell culture. Oxidized complexes caused no cytotoxicity even at high polymer concentrations. Compared to non-oxidized complexes used at subtoxic doses, oxidized complexes mediated significantly more GFP expression. A key strength of this approach is its simplicity as it involves only simple mixing of solutions. This strategy promises to further realize the potential of using PEI for the delivery of nucleic acids or other cargos.


Barriers and facilitators to diabetes self-management: perspectives of older community dwellers and health professionals in China.
Little is known about self-management among people with Type 2 diabetes living in mainland China. Understanding the experiences of this target population is needed to provide socioculturally relevant education to effectively promote self-management. The aim of this study was to explore perceived barriers and facilitators to diabetes self-management for both older community dwellers and health professionals in China. Four focus groups, two for older people with diabetes and two for health professionals, were conducted. All participants were purposively sampled from two communities in Shanghai, China. Six barriers were identified: overdependence on but dislike of western medicine, family role expectations, cuisine culture, lack of trustworthy information sources, deficits in communication between clients and health professionals, and restriction of reimbursement regulations. Facilitators included family and peer support, good relationships with health professionals, simple and practical instruction and a favourable community environment. The findings provide valuable information for diabetes self-management intervention development in China, and have implications for programmes tailored to populations in similar sociocultural circumstances.


CENP-A Subnuclear Localization Pattern as Marker Predicting Curability by Chemoradiation Therapy for Locally Advanced Head and Neck Cancer Patients.
Effective biomarkers predictive of the response to treatments are key for precision medicine. This study identifies the staining pattern of the centromeric histone 3 variant, CENP-A, as a predictive biomarker of locoregional disease curability by chemoradiation therapy. We compared by imaging the subnuclear distribution of CENP-A in normal and tumoral tissues, and in a retrospective study in biopsies of 62 locally advanced head and neck squamous cell carcinoma (HNSCC) patients treated by chemoradiation therapy. We looked for predictive factors of locoregional disease control and patient's survival, including CENP-A patterns, Ki67, HPV status and anisokaryosis. In different normal tissues, we reproducibly found a CENP-A subnuclear pattern characterized by CENP-A clusters both localized at the nuclear periphery and regularly spaced. In corresponding tumors, both features are lost. In locally advanced HNSCC, a specific CENP-A pattern identified in pretreatment biopsies predicts definitive locoregional disease control after chemoradiation treatment in 96% (24/25) of patients (OR = 17.6 CI 95% [2.6; 362.8], p = 0.002), independently of anisokaryosis, Ki67 labeling or HPV status. The characteristics of the subnuclear pattern of CENP-A in cell nuclei revealed by immunohistochemistry could provide an easy to use a reliable marker of disease curability by chemoradiation therapy in locally advanced HNSCC patients.


Calcium entry blockade and adrenergic vascular reactivity in hypertensives: differences between nicardipine and diltiazem.
The interference of nicardipine and diltiazem infused into the brachial artery at systemically ineffective rates, with the forearm vascular response to graded exogenous norepinephrine, was evaluated in hypertensive patients. Nicardipine (1 and 3 micrograms/dl forearm tissue/min in both absence and presence of propranolol) increased forearm blood flow (venous plethysmography) and antagonized dose dependently the vasoconstrictor effect of norepinephrine, suggesting that functional alpha-antagonism may participate in the vasodilating and possibly the antihypertensive effect of the drug. On the contrary, no antagonism but rather potentiation of the responses to norepinephrine occurred after diltiazem (0.5 and 1 microgram/dl forearm tissue/min). Because intra-arterial propranolol abolished that potentiating action of the drug, whereas the local vasodilation to isoproterenol was clearly reduced, diltiazem probably interfered with beta-adrenergic receptor-mediated vasorelaxing mechanisms in human forearm arterioles. The data further stress the heterogeneity of calcium entry blockers in humans.


Dancing as a psychosocial intervention in care homes: a systematic review of the literature.
There is a need to find meaningful and engaging interventions to improve mood and behaviour for residents of care homes. The demand on care staff might diminish opportunities for them to encourage these activities. Staff anecdotal information attests that dancing as an activity improves mood in residents and staff. Hence, the importance of investigating what dancing brings to the care home social environment. To provide a systematic review of the evidence from studies related to dancing interventions for older people with dementia living in care homes. Electronic databases were searched. Previous reviews were also included, and recognised experts were consulted up to January 2012. Inclusion criteria considered study methodology and evidence that the impact of the dance intervention had been measured. Ten studies were identified that satisfied the inclusion criteria: seven qualitative and three quantitative. Studies used different approaches such as therapeutic dance, dance movement therapy, dance therapy, social dancing and psychomotor dance-based exercise. There was evidence that problematic behaviours decreased; social interaction and enjoyment in both residents and care staff improved. A few adverse effects were also acknowledged. The evidence on the efficacy of dancing in care homes is limited in part owing to the methodological challenges facing such research. This review aims to raise awareness of the possibility of implementing dance work as an engaging activity in care homes. We shall also consider options for future dance work research as a means to encourage relationships and sensory stimulation for both residents and staff.


New Therapies in Melanoma: Current Trends, Evolving Paradigms, and Future Perspectives.
Melanoma is an aggressive skin cancer with increasing incidence and mortality worldwide. For many years the therapeutic strategies were limited to surgery, radiotherapy, and chemotherapy. Recent advances in immunology and cancer biology have led to the discovery and development of novel therapeutics, such as immune checkpoint inhibitors (ICIs) and targeted therapies, which have revolutionized the clinical care of patients with metastatic melanoma. Despite recent successes with ICIs, many melanoma patients do not experience long-term benefits from ICI therapies, highlighting the need for alternative treatments with novel targets such as lymphocyte-activated gene 3 (LAG-3). In this review, we explore new therapeutic agents and novel combinations that are being tested in early-phase clinical trials. We discuss newer promising tools such as nanotechnology to develop nanosystems that act as drug carriers and/or light absorbents to potentially improve therapy outcomes. Finally, we also highlight challenges such as management after resistance and intervention with novel immunotherapies and the lack of predictive biomarkers to stratify patients to targeted treatments after primary treatment failure.


How to escape the cancer attractor: rationale and limitations of multi-target drugs.
The increasingly evident limitations of target-selective cancer therapy has stimulated a flurry of ideas for overcoming the development of resistance and recurrence - the near universal reason for therapy failure from which target-selective drugs are not exempt. A widely proposed approach to conquer therapy resistance is to depart from the myopic focus on individual causal pathways and instead target multiple nodes in the cancer cell's gene regulatory network. However, most ideas rely on a simplistic conceptualization of networks: utilizing solely their topology and treating it as a display of causal interactions, while ignoring the integrated dynamics in state space. Here, we review the more encompassing formal framework of global network dynamics in which cancer cells, like normal cell types, are high-dimensional attractor states. Then therapy is represented by the network perturbation that will promote the exit from such cancer attractors and reentering a normal attractor. We show in this qualitative and accessible discussion how the idea of a quasi-potential landscape and the theory of least-action-path offer a new formal understanding for computing the set of network nodes (molecular targets) that need to be targeted in concert in order to exit the cancer attractor. But targeting cancer cells based on the network configuration of an "average" cancer cell, however precise, may not suffice to eradicate all tumor cells because of the dynamic non-genetic heterogeneity of cancer cell populations that makes them moving targets and drives the replenishment of the cancer attractor with surviving, non-responsive cells from neighboring abnormal attractors.


CECR1-mediated cross talk between macrophages and vascular mural cells promotes neovascularization in malignant glioma.
Glioblastomas (glioblastoma multiforme, GBM) are most malignant brain tumors characterized by profound vascularization. The activation of macrophages strongly contributes to tumor angiogenesis during GBM development. Previously, we showed that extracellular adenosine deaminase protein Cat Eye Syndrome Critical Region Protein 1 (CECR1) is highly expressed by M2-like macrophages in GBM where it defines macrophage M2 polarization and contributes to tumor expansion. In this study, the effect of CECR1 in macrophages on tumor angiogenesis was investigated. Immunohistochemical evaluation of GBM tissue samples showed that the expression of CECR1 correlates with microvascular density in the tumors, confirming data from the TCGA set. In a three-dimensional co-culture system consisting of human pericytes, human umbilical vein endothelial cells and THP1-derived macrophages, CECR1 knockdown by siRNA and CECR1 stimulation of macrophages inhibited and promoted new vessel formation, respectively. Loss and gain of function studies demonstrated that PDGFB mRNA and protein levels in macrophages are modulated by CECR1. The proangiogenic properties of CECR1 in macrophages were partially mediated via paracrine activation of pericytes by PDGFB-PDGFRβ signaling. CECR1-PDGFB-PDGFRβ cross-activation between macrophages and pericytes promoted pericyte migration, shown by transwell migration assay, and enhanced expression and deposition of periostin, a matrix component with proangiogenic properties. CECR1 function in (M2-like) macrophages mediates cross talk between macrophages and pericytes in GBM via paracrine PDGFB-PDGFRβ signaling, promoting pericyte recruitment and migration, and tumor angiogenesis. Therefore, CECR1 offers a new portent target for anti-angiogenic therapy in GBM via immune modulation.