Raloxifene versus continuous combined estrogen/progestin therapy: densitometric and biochemical effects in healthy postmenopausal Taiwanese women.
We treated 116 healthy postmenopausal women (age 47-66 years, mean 57 years) in Taiwan with either raloxifene (RLX) 60 mg (n = 92) or 0.625 mg conjugated equine estrogen plus 5 mg medroxyprogesterone acetate (CCEP, n = 24) daily for 12 months in a randomized, double-masked, active-controlled fashion. The results showed that both regimens increased bone mineral density (BMD) at hip sites (means: RLX 2.5-4.9%, CCEP 4.6-7.9%, all p<0.005 compared with baseline), and the difference between the two regimens was not significant. The spinal BMD increased significantly in both groups (1.4% with RLX and 6.0% with CCEP, both p<0.01), and more with CCEP (p<0.003). Osteocalcin levels and urinary type I collagen C-telopeptide/creatinine ratios decreased significantly in both regimens, but the decreases were significantly larger with CCEP. Compared with baseline, both RLX and CCEP decreased the total cholesterol (median 4.9% and 8.6% respectively, p<0.001) and LDL-cholesterol (median 11% and 19% respectively, p<0.001), and increased HDL-cholesterol (median 8.6% and 10.7% respectively, p<0.01). Both regimens increased triglyceride levels (median 3.2% and 18.9% respectively, both p<0.05), although to a lesser extent with RLX than with CCEP (p<0.05). Only 3 subjects (3.3%) reported vaginal bleeding in the RLX group, as compared with 31% (7/22) with CCEP (p<0.05). We conclude that in healthy, postmenopausal Taiwanese women, RLX 60 mg given daily has favorable results in BMD, bone turnover and serum lipids, although the dosage we used showed a potency less than that of conventional CCEP.


γ-Oryzanol suppresses cell apoptosis by inhibiting reactive oxygen species-mediated mitochondrial signaling pathway in H2O2-stimulated L02 cells.
γ-Oryzanol, a mixture of ferulic acid esters of plant sterols and triterpene alcohols existed in rice bran oil, can ameliorate lipid metabolism and enhance antioxidant activity. In this study, we used hydrogen peroxide (H2O2)-induced injury in human hepatic L02 cells to investigate the mechanisms involved in the hepatoprotective activity of γ-oryzanol. The injuries produced by H2O2 in L02 cells include increased levels of malondialdehyde (MDA) and intracellular reactive oxygen species (ROS), decreased activities of superoxide dismutase (SOD) and catalase (CAT), loss of mitochondrial membrane potential (MMP), increased protein expressions of caspase-9 and caspase-3, and induced apoptosis. Pretreatment with γ-oryzanol enhanced the ROS scavenging activity of endogenous antioxidant enzymes and decreased lipid peroxidation in H2O2 treated cells. Moreover, pretreatment with γ-oryzanol inhibited H2O2-induced apoptosis by restoring MMP, upregulating the expression ratio of Bcl-2/Bax, and inhibiting the activation of caspase-9 and caspase-3. These findings show that γ-oryzanol can prevent H2O2-induced apoptosis by suppressing intracellular accumulation of ROS and impeding ROS-activated mitochondrial apoptotic pathway.


Familial human hypodontia--is it all in the genes?
The congenital absence of teeth is one of the commonest developmental abnormalities seen in human populations. Familial hypodontia or oligodontia represents an absence of varying numbers of primary and/or secondary teeth as an isolated trait. While much progress has been made in understanding the developmental basis of tooth formation, knowledge of the aetiological basis of inherited tooth loss remains poor. The study of mouse genetics has uncovered a large number of candidate genes for this condition, but mutations in only three have been identified in human pedigrees with familial hypodontia or oligodontia: MSX1, PAX9 and AXIN2. This suggests that these conditions may represent a more complex multifactorial trait, influenced by a combination of gene function, environmental interaction and developmental timing. Completion of the human genome project has made available the DNA sequence of the collected human chromosomes, allowing the localisation of all human genes and, ultimately, determination of their function. Therefore it is likely that our understanding of this complex developmental process will continue to improve, not only during normal development but also when things go wrong.


Immunostained cathepsins B and L correlate with depth of invasion and different metastatic pathways in early stage gastric carcinoma.
With the recent development of minimal treatment for early stage gastric carcinoma, identifying specific indicators of the metastatic potential of primary tumors has become more important. Cathepsin B and cathepsin L, both lysosomal cysteine proteases, degrade the extracellular matrix during tumor progression. Although many studies have shown their relation to human cancer progression, little is known about their roles in the early stage. The clinicopathologic significance of cathepsins was therefore studied in early stage gastric carcinoma. Expression of both cathepsins was studied immunohistochemically in 51 tissue specimens from gastric carcinomas that invaded the submucosal layer or muscularis propria. The relation between their expression and clinicopathologic factors was analyzed. Both cathepsins were expressed at higher levels in tumors that invaded the muscularis propria than in those within the submucosa (P < 0.05). In addition, tumors with lymphatic invasion showed higher cathepsin B expression than those without it (P < 0.05), whereas tumors with venous invasion showed higher cathepsin L expression than those without it (P < 0.05). No other clinicopathologic factors correlated with expression of either cathepsin. Tumors with overexpression of cathepsins have powerful potential for invasiveness in the early stage of gastric carcinoma. Moreover, the authors hypothesize that cathepsins may be one of the determinants of the metastatic route. To the authors' knowledge, this is the first report on specific proteases concerning the mode of metastasis, and the results of this study suggest that therapeutic strategies for early stage gastric carcinoma might need to be changed according to the status of cathepsins.


Danger is in the eyes of the beholder: The effect of visible and invisible affective faces on the judgment of social interactions.
Previous studies demonstrated that observation of facial expressions can modulate threat detection while looking at neutral or emotion-related scenes. Similarly, stimuli presented outside conscious awareness could influence social judgments of neutral novel stimuli. The two-fold aim of this study was: i) to evaluate whether observation of seen emotional faces could affect the judgment of social interactions without contextual cues (visible prime condition), and ii) whether this effect could also emerge when the emotional faces were made not visible by means of continuous flash suppression (invisible prime condition). We found that both seen and unseen faces are able to affect the judgment of ambiguous social interactions although this effect was particularly evident when affective faces were clearly visible. The present findings supported the idea that both conscious and unconscious processing of emotional faces have an important role in modulating perceivers' affective state and their judgment of social interactions.


Targeting T cells responsive to the priming epitope prevent the relapsing phase of experimental autoimmune encephalomyelitis.
Upon recovery from the initial episode of experimental autoimmune encephalomyelitis (EAE), virtually all SJL mice develop relapsing/remitting episodes of disease. These relapses may occur due to the reactivation of memory T cells initially stimulated as part of the disease-inducing protocol or naïve T-cell populations stimulated by distinct encephalitogens derived from the inflammatory disease process (epitope spread). We have used encephalitogen-specific non-linear peptide octamers to modify the course of relapsing EAE (rEAE) in SJL mice immunized with an oliogodendrocyte-specific protein peptide (OSP 55-71). Our studies show that the peptide-octamers, which target the T cells stimulated by the priming encephalitogen, but not other candidate encephalitogens, prevent rEAE.


Investigations into the association between cervicomedullary neuroschisis and mirror movements in patients with Klippel-Feil syndrome.
Our purpose was to investigate the association between cervicomedullary neuroschisis and mirror movements in patients with Klippel-Feil syndrome (KFS). We conducted a retrospective analysis of 23 patients with KFS who were seen at our institution during a 10-year period. Sixteen of the 23 patients had undergone adequate axial view cross-sectional imaging of the upper cervical spine. The degree of neuroschisis was assessed for each patient, using an objective scoring system. Twelve patients were evaluated for the presence or absence of mirror movements. A high percentage of female patients with KFS was noted (17 [74%] of 23 patients). Adequate cross-sectional images were available for 16 of the 23 patients, six (38%) of whom had some form of cervicomedullary neuroschisis. Five of the six patients had been clinically evaluated, and all were shown to have mirror movements. One patient with Chiari II malformation, which obscured evaluation for neuroschisis, also had mirror movements. Of the remaining nine patients without cervicomedullary neuroschisis, six were evaluated, and none of the six had mirror movements. A review of the theoretical neuroanatomic basis of mirror movements is presented herein, and neurosurgical management concerns for patients with KFS are discussed. A strong association exists between cervicomedullary neuroschisis and mirror movements in cases of KFS. Screening of patients with mirror movements may help identify clinically unsuspected KFS and may also help stratify risk within this patient population, identifying patients who might benefit from early neurosurgical intervention.


Glucocorticoid-induced osteoporosis: 2019 concise clinical review.
Glucocorticoids remain widely used for many medical conditions, and fractures are the most serious common adverse event related to long-term glucocorticoid use. Glucocorticoid-induced osteoporosis (GIOP) develops in a time- and dose-dependent manner, but even at low doses, an increased risk of fragility fracture may be observed even within the first month of treatment. GIOP is mediated by multiple pathophysiologic mechanisms resulting in an inhibition of bone formation and an increase in bone resorption. The clinical assessment of GIOP has potential pitfalls since dual-energy X-ray absorptiometry (DXA) may underestimate the risk of fracture in patients treated with glucocorticoids. Many national organizations have developed guidelines for assessing fracture risk and treating patients with, or at risk for, GIOP. These groups advocate both antiresorptive agents and bone-forming agents based predominately on their efficacy in improving bone mineral density. Oral bisphosphonates are generally the first-line therapy for GIOP in most patients due to their proven efficacy, good safety, and low cost. For those patients at greater risk of fracture, teriparatide should be considered earlier, based on its ability to significantly reduce vertebral fractures when compared with alendronate. GIOP remains a major public health concern that is at least partially preventable with current and potential future therapeutic options.


Beyond the red cell: pegylation of other blood cells and tissues.
Immunological recognition of allogeneic tissue is of critical concern in transfusion and transplantation medicine. While the major emphasis of our work on the immunocamouflage of cells has been focused on the erythrocyte, we have extended this research beyond the red blood cell (RBC) to other tissues. Our studies from blood transfusion (i.e., a specialized form of cellular transplantation) suggest that covalent modification of cells and tissues with methoxypoly(ethylene glycol) mPEG can significantly diminish immunologic recognition of other allogeneic tissues and, furthermore, may enhance the induction of tolerance. The mechanisms underlying the mPEG-mediated immunocamouflage of alloantigens is the global camouflaging of antigenic sites, membrane surface charge and the attenuation of receptor-ligand and cell-cell interactions. As a consequence of the immunocamouflage imparted by the grafted mPEG, weak costimulation of alloreactive T cells is observed which subsequently induces apoptosis of these reactive cells. As a result of this clonal deletion, a pro-tolerance state is induced. The potency of immunocamouflage is readily observed in in vivo murine models of transfusion-associated graft versus host disease. Furthermore, initial studies on the in vivo transplantation of pegylated rat and murine pancreatic islets have demonstrated that mPEG-derivatization does not impair the finely tuned signaling necessary for glucose homeostasis. Finally, in contrast to the pharmacological inhibition of the immune response by agents such as cyclosporine, mPEG-mediated immunocamouflage directly attenuates the inherent antigenicity and immunogenicity of the donor tissue itself while leaving the recipient a fully competent immune system.


The Dual Roles of Phenylenediamines: Using their Voltammetric Behavior to Measure the Hydrogen Donor and Acceptor Abilities of Alcohols in Acetonitrile.
Cyclic voltammetry experiments on 2,3,5,6-tetramethyl-1,4-phenylenediamine (P) in acetonitrile in the presence of varying concentrations of alcohols indicate that the oxidized forms of the compound (P.+ and P2+ ) interact with the alcohols through a hydrogen-bonding mechanism where P.+ and P2+ act as the hydrogen donor and the alcohols act as acceptors. However, the neutral form (P) largely acts as a hydrogen acceptor but only for strong hydrogen donors that do not undergo proton-transfer reactions with the phenylenediamine. These results were ascertained based on measuring the difference in potential of the two one-electron transfer reactions (ΔEPox(1, 2) =|EPox(1) -EPox(2) |) in the oxidative electrochemistry of P, which thereby allows a simple measure of relative hydrogen bonding strengths.


A study on withdrawal related haemodynamic response in chronic propranolol treated conscious rats.
Single dose of propranolol hydrochloride (5 mg/kg, i.p.) caused significant fall in heart rate (HR) but not in systolic blood pressure (SBP) in normotensive conscious rats. Multiple doses of propranolol (5 mg/kg, i.p., twice-a-day for 5 wk) caused significant fall in both HR and SBP at 2 wk and 4 wk in normotensive conscious rats. Sudden withdrawal of propranolol at 5 wk caused a significant blood pressure upswing and tachycardia between 12-24 h followed by normalization of both blood pressure and heart rate. The study documents a possible model of rebound hypertension in normotensive conscious rats.


Role of CT and MRI in the design and development of orthopaedic model using additive manufacturing.
To study the role of Computed tomography (CT) and Magnetic resonance imaging (MRI) for design and development of orthopaedic model using additive manufacturing (AM) technologies. A significant number of research papers in this area are studied to provide the direction of development along with the future scope. Briefly discussed various steps used to create a 3D model by Additive Manufacturing using CT and MRI scan. These scanning technologies are used to produce medical as well as orthopaedic implants by using AM technologies. The images so produced are exported in different software like OsiriX Imaging Software, 3D slicer, Mimics, Magics, 3D doctor and InVesalius to produce a 3D digital model. Various criteria's achieved by CT and MRI scan for design and development of orthopaedic implant using additive manufacturing are also discussed briefly. AM model created by this process show exact shape, size, dimensions, textures, colour and features. AM technologies help to convert the digital model into a 3D physical object, thereby improving the understanding of patient anatomy for treatment as well as for educational purpose. These scanning technologies have various applications to enhance the AM in the field of orthopaedic. In orthopaedic every patient model is a customised unit, sourced from the individual patient. 3D CAD data captured by these scanning technologies are directly exported in standard triangulate language (STL) format for printing by AM technologies. Crossestion of the physical model fabricated by this process shows a patient's anatomy if the model prepared by using the bone-like material.


Assessment of the Severity of Ischaemia and the Outcomes of Revascularisation in Peripheral Arterial Disease Patients Based on the Skin Microcirculatory Response to a Thermal Load Test.
This study investigated the skin microcirculatory response to a thermal load test using a laser Doppler flowmetry device to evaluate the severity of limb ischaemia and the outcomes of revascularisation in patients with peripheral arterial disease (PAD). A total of 34 PAD patients (39 limbs) including 17 critical limb ischaemia (CLI) patients (21 limbs) who underwent revascularisation were enrolled. The skin microcirculation of the dorsal side of the affected foot was investigated for 15 minutes after local heating. The tests were performed both before and after revascularisation, and several parameters gleaned from the microcirculatory fluctuations were analysed and compared with the ankle brachial pressure index and the transcutaneous oxygen tension (tcPO2) values. Among the parameters, significant differences were observed between the CLI patients and patients with claudication with regard to the increasing phase time (Tinc), the difference in the perfusion values at the onset and the peak of the transient increase in blood perfusion (PΔ), the slope of the transient increase in blood perfusion (Sin), and the slope of the decrease in blood perfusion after the peak (Sde). In CLI patients, the PΔ, Sin, and Sde values increased significantly after revascularisation. In the patients with claudication, the changes in the parameters after revascularisation were not statistically significant. The Sde showed the most statistically significant correlation with the tcPO2 value (ρ .759, p < .001). Thermal load testing can be used to evaluate the severity of limb ischaemia in patients with PAD.


The current status of phage therapy and its advancement towards establishing standard antimicrobials for combating multi drug-resistant bacterial pathogens.
Phage therapy; a revived antimicrobial weapon, has great therapeutic advantages with the main ones being its ability to eradicate multidrug-resistant pathogens as well as selective toxicity, which ensures that beneficial microbiota is not harmed, unlike antibiotics. These therapeutic properties make phage therapy a novel approach for combating resistant pathogens. Since millions of people across the globe succumb to multidrug-resistant infections, the implementation of phage therapy as a standard antimicrobial could transform global medicine as it offers greater therapeutic advantages than conventional antibiotics. Although phage therapy has incomplete clinical data, such as a lack of standard dosage and the ideal mode of administration, the conducted clinical studies report its safety and efficacy in some case studies, and therefore, this could lessen the concerns of its skeptics. Since its discovery, the development of phage therapeutics has been in a smooth progression. Concerns about phage resistance in populations of pathogenic bacteria are raised when bacteria are exposed to phages. Bacteria can use restriction-modification, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein (Cas) defense, or mutations in the phage receptors to prevent phage invasion. Phage resistance, however, is often costly for the bacteria and may lead to a reduction in its virulence. The ongoing competition between bacteria and phage, on the other hand, ensures the emergence of phage strains that have evolved to infect resistant bacteria. A phage can quickly adapt by altering one or more aspects of its mode of infection, evading a resistance mechanism through genetic modifications, or directly thwarting the CRISPR-Cas defense. Using phage-bacterium coevolution as a technique could be crucial in the development of phage therapy as well. Through its recent advancement, gene-editing tools such as CRISPR-Cas allow the bioengineering of phages to produce phage cocktails that have broad spectrum activities, which could maximize the treatment's efficacy. This review presents the current state of phage therapy and its progression toward establishing standard medicine for combating antibiotic resistance. Recent clinical trials of phage therapy, some important case studies, and other ongoing clinical studies of phage therapy are all presented in this review. Furthermore, the recent advancement in the development of phage therapeutics, its application in various sectors, and concerns regarding its implementation are also highlighted here. Phage therapy has great potential and could help the fight against drug-resistant bacterial pathogens.


Stimulus responsive nanogel with innate near IR fluorescent capability for drug delivery and bioimaging.
A brighter, non toxic and biocompatible optical imaging agent is one of the major quests of biomedical research. Here in, we report a photoluminescent comacromer [PEG-poly(propylene fumarate)-citric acid-glycine] and novel stimulus (pH) responsive nanogel endowed with excitation wavelength dependent fluorescence (EDF) for combined drug delivery and bioimaging applications. The comacromer when excited at different wavelengths in visible region from 400nm to 640nm exhibits fluorescent emissions from 510nm to 718nm in aqueous condition. It has high Stokes shift (120nm), fluorescent lifetime (7 nanoseconds) and quantum yield (50%). The nanogel, C-PLM-NG, prepared with this photoluminescent comacromer and N,N-dimethyl amino ethylmethacrylate (DMEMA) has spherical morphology with particle size around 100nm and 180nm at pH 7.4 (physiological) and 5.5 (intracellular acidic condition of cancer cells) respectively. The studies on fluorescence characteristics of C-PLM NG in aqueous condition reveal large red-shift with emissions from 523nm to 700nm for excitations from 460nm to 600nm ascertaining the EDF characteristics. Imaging the near IR emission with excitation at 535nm was accomplished using cut-off filters. The nanogel undergoes pH responsive swelling and releases around 50% doxorubicin (DOX) at pH 5.5 in comparison with 15% observed at pH 7.4. The studies on in vitro cytotoxicity with MTT assay and hemolysis revealed that the present nanogel is non-toxic. The DOX-loaded C-PLM-NG encapsulated in Hela cells induces lysis of cancer cells. The inherent EDF characteristics associated with C-PLM NG enable cellular imaging of Hela cells. The studies on biodistribution and clearance mechanism of C-PLM-NG from the body of mice reveal bioimaging capability and safety of the present nanogel. This is the first report on a polymeric nanogel with innate near IR emissions for bioimaging applications.


In suspected PE, YEARS rule + D-dimer was noninferior to D-dimer alone to rule out PE.
Freund Y, Chauvin A, Jimenez S, et al. Effect of a diagnostic strategy using an elevated and age-adjusted D-dimer threshold on thromboembolic events in emergency department patients with suspected pulmonary embolism: a randomized clinical trial. JAMA. 2021;326:2141-9. 34874418.


High frequency of cerebrospinal fluid autoantibodies in COVID-19 patients with neurological symptoms.
COVID-19 intensive care patients can present with neurological syndromes, usually in the absence of SARS-CoV-2 in cerebrospinal fluid (CSF). The recent finding of some virus-neutralizing antibodies cross-reacting with brain tissue suggests the possible involvement of specific autoimmunity. Blood and CSF samples from eleven critically ill COVID-19 patients presenting with unexplained neurological symptoms including myoclonus, oculomotor disturbance, delirium, dystonia and epileptic seizures, were analyzed for anti-neuronal and anti-glial autoantibodies. Using cell-based assays and indirect immunofluorescence on unfixed murine brain sections, all patients showed anti-neuronal autoantibodies in serum or CSF. Antigens included intracellular and neuronal surface proteins, such as Yo or NMDA receptor, but also various specific undetermined epitopes, reminiscent of the brain tissue binding observed with certain human monoclonal SARS-CoV-2 antibodies. These included vessel endothelium, astrocytic proteins and neuropil of basal ganglia, hippocampus or olfactory bulb. The high frequency of autoantibodies targeting the brain in the absence of other explanations suggests a causal relationship to clinical symptoms, in particular to hyperexcitability (myoclonus, seizures). Several underlying autoantigens and their potential molecular mimicry with SARS-CoV-2 still await identification. However, autoantibodies may already now explain some aspects of multi-organ disease in COVID-19 and can guide immunotherapy in selected cases.


Acute hyperglycaemia following thermal injury: friend or foe?
Hyperglycaemia and insulin resistance are common in severely burned patients, even if they have not previously had diabetes. Conventionally, hyperglycaemia is considered a part of the hypermetabolic stress response and blood glucose levels up to 215 mg/dl are tolerated before insulin therapy is initiated. Recent studies suggest that hyperglycaemia and insulin resistance are harmful and that correcting blood glucose to normal levels with insulin might improve the prognosis significantly. The purpose of this clinical study was to evaluate blood glucose levels in severely burned patients with conventional management and to analyse the association between early hyperglycaemia and clinical outcome. Clinical, prospective, descriptive study. Thirty seven severely burned adults (>25% total body surface area). Hyperglycaemia was treated according to conventional clinical practice. This included the infusion of insulin based on a blood glucose level >215 mg/dl and the maintenance of the glucose level between 180 and 200 mg/dl. Measurements of whole-blood glucose were performed at 8, 16, 24, 36 and 48 h after the thermal injury. Additional measurements were performed if indicated. A total of 185 measurements were obtained and significant elevations of blood glucose levels (>140 mg/dl) were found in 108 (64%) of the measurements. Peak blood glucose values exceeded 140 mg/dl in all but three of the patients; however, only 17 patients received insulin treatment during the shock period. The inadequacy of the insulin treatment is shown by the mean glucose values, which exceeded 200 mg/dl in 27% of the patients. Despite a non-significant difference in the extent of burn (P=0.055), patients who died showed significantly higher maximum glucose values than patients who survived the thermal injury (P<0.05). Even though not statistically significant, blood glucose control was poorer in patients who later developed sepsis or acute renal failure (P>0.05). No correlation was found between burned surface area (TBSA) and mean plasma glucose levels during the first 48 h of resuscitation (r=0.12). Hyperglycaemia is very frequent during the resuscitation period of thermal injury and current guidelines for insulin therapy are inadequate to correct plasma glucose to normal levels. As an association between early hyperglycaemia and subsequent mortality seems to exist, more aggressive manoeuvres to reduce blood glucose may be warranted in this group of patients.


Bayesian model of disease progression in mucopolysaccaridosis IIIA.
Mucopolysaccaridosis IIIA (MPS IIIA) is a rare genetic disease that afflicts children and leads to neurocognitive degeneration. We develop a Bayesian disease progression model (DPM) of MPS IIIA that characterizes the pattern of cognitive growth and decline in this disease. The DPM is a repeated measures model that incorporates a nonlinear developmental trajectory and shape-invariant random effects. This approach quantifies the pattern of cognitive development in MPS IIIA and addresses differences in biological age, length of follow-up, and clinical outcomes across natural history subjects. The DPM can be used in clinical trials to estimate the percent slowing in disease progression for treatment relative to natural history. Simulations demonstrate that the DPM provides substantial improvements in power relative to alternative analyses.


Sonographic evaluation of neonates with early-stage necrotizing enterocolitis.
Necrotizing enterocolitis (NEC) is the most common gastrointestinal emergency in neonatal intensive care units. Ultrasonographic findings in early-stage NEC have not been described. To assess the diagnostic value of ultrasonography for the diagnosis and monitoring of patients with NEC. We evaluated the sonographic findings of early stages of NEC in 40 neonates who were clinically diagnosed with NEC when they were 2-28 days old. Their average gestational age was 32 weeks, and their mean weight was 1,850 g. All of the patients showed signs of bowel distention on abdominal radiography, with no evidence of pneumatosis intestinalis. We performed bowel sonography in all patients (n = 40), as well as in ten healthy neonates who served as a control group. The studies were conducted with a 10-MHz linear transducer from February 2003 to January 2004. We evaluated the echogenicity of the bowel wall, involved region, ascites, and portal venous gas at both initial and follow-up examinations. We divided the patients into two groups according to the bowel wall echogenicity pattern, group I with echogenic dots in the bowel wall and group II with dense granular echogenicities in the bowel wall. In order to identify any correlations between the ultrasonography and clinical findings, we evaluated the duration of parenteral feeding (NPO) in each group and compared two groups by means of a statistical analysis (Mann-Whitney test). All of the neonates in the control group (n = 10) presented normal bowel wall echogenicity; the patients with NEC presented echogenic dots in 16 patients (40%) and dense granular echogenicities in 24 patients (60%). Portal venous gas was absent in all patients. On the follow-up examinations, the echogenicity of the bowel wall and ascites decreased in 37 patients (93%). The duration of NPO was 11.1 +/- 6.6 days in group I and 16.5 +/- 7.2 days in group II (P < 0.05). Echogenic dots or dense granular echogenicities in the bowel wall can be seen in patients with early-stage NEC. Bowel sonography can be helpful for the early diagnosis and monitoring of patients with NEC.


Persistence of functional memory B cells recognizing SARS-CoV-2 variants despite loss of specific IgG.
Although some COVID-19 patients maintain SARS-CoV-2-specific serum immunoglobulin G (IgG) for more than 6 months postinfection, others eventually lose IgG levels. We assessed the persistence of SARS-CoV-2-specific B cells in 17 patients, 5 of whom had lost specific IgGs after 5-8 months. Differentiation of blood-derived B cells in vitro revealed persistent SARS-CoV-2-specific IgG B cells in all patients, whereas IgA B cells were maintained in 11. Antibodies derived from cultured B cells blocked binding of viral receptor-binding domain (RBD) to the cellular receptor ACE-2, had neutralizing activity to authentic virus, and recognized the RBD of the variant of concern Alpha similarly to the wild type, whereas reactivity to Beta and Gamma were decreased. Thus, differentiation of memory B cells could be more sensitive for detecting previous infection than measuring serum antibodies. Understanding the persistence of SARS-CoV-2-specific B cells even in the absence of specific serum IgG will help to promote long-term immunity.


Speciation of ionic alkyllead in potable water and soil.
Various potable water and soil samples have been analyzed for tri- and dialkyllead compounds using a sensitive speciation procedure based on diethyldithiocarbamate extraction, Grignard derivatization and gas chromatography-atomic absorption spectrometry. The species are generally present as ultra-trace contaminants, and their abundance is critically discussed. In addition, a degradation study of ionic alkyllead in ambient matrices is presented.


[Effect of coronary revascularization in patients with ischemic cardiomyopathy who satisfy viability criteria on gated-SPECT assessment of myocardial perfusion].
Study on the effects of coronary revascularization and medical treatment on the prognosis of patients with ischemic cardiomyopathy who satisfy myocardial viability criteria on gated single-photon emission computed tomography (SPECT) assessment of myocardial perfusion. In total, 140 consecutive patients with ischemic cardiomyopathy were studied at rest using gated-SPECT and technetium-labeled contrast agents. During a mean follow-up period of 2.3 years after gated-SPECT, the rate of cardiac death in patients who underwent coronary revascularization (n=50) was 16% compared with 26.7% in those who received medical treatment (n=90). Thus, coronary revascularization had a protective effect (hazard ratio = 0.42; 95% confidence interval, 0.17-1.02) in patients with ischemic cardiomyopathy who satisfied myocardial viability criteria.


Factors that Associate Japanese University Students' Use of Rear Seat Belts on General Roads and Expressways.
This study clarifies the prevalence of rear seat belt use among university students and explores the factors that influence seat belt use on general roads and expressways. Questionnaires were distributed to 1,091 students who attended health education lectures: 893 students (81.9%) agreed to participate and met the criteria; among them, 660 (73.9%) provided complete responses. While one-third (33.0%) of rear seat passengers "always" used seat belts on general traffic roads, three-fourths (73.2%) "always" used them on expressways. Multivariate analysis revealed that being male and the perception of obligation and recognition of the benefits of rear seat belt use were associated with use on general roads, whereas the possession of a driver's license and recognition of benefits were associated with use on expressways. Finally, we suggest that educational programs on seat belt use be developed and delivered to general road and expressway users to promote seat belt use.


Chronic treatment with corticosterone increases the number of tyrosine hydroxylase-expressing cells within specific nuclei of the brainstem reticular formation.
Cushing's syndrome is due to increased glucocorticoid levels in the body, and it is characterized by several clinical alterations which concern both vegetative and behavioral functions. The anatomical correlates of these effects remain largely unknown. Apart from peripheral effects induced by corticosteroids as counter-insular hormones, only a few reports are available concerning the neurobiology of glucocorticoid-induced vegetative and behavioral alterations. In the present study, C57 Black mice were administered daily a chronic treatment with corticosterone in drinking water. This treatment produces a significant and selective increase of TH-positive neurons within two nuclei placed in the lateral column of the brainstem reticular formation. These alterations significantly correlate with selective domains of Cushing's syndrome. Specifically, the increase of TH neurons within area postrema significantly correlates with the development of glucose intolerance, which is in line with the selective control by area postrema of vagal neurons innervating the pancreas. The other nucleus corresponds to the retrorubral field, which is involved in the behavioral activity. In detail, the retrorubral field is likely to modulate anxiety and mood disorders, which frequently occur following chronic exposure to glucocorticoids. To our knowledge, this is the first study that provides the neuroanatomical basis underlying specific symptoms occurring in Cushing's syndrome.


Inter-species sequence diversity in the replication initiation region of Paramecium mitochondrial DNA.
Mitochondrial DNA from Paramecium aurelia is a linear molecule. Replication is initiated at a unique cross-linked molecular terminus. During replication dimer length molecules, consisting of two head-to-head monomers, are generated. We have cloned the head-to-head dimer initiation region from five different species and several stocks (or races) within species and determined its DNA sequence. For all species, this dimer initiation region consists of a central non-palindromic sequence containing almost exclusively A and T, arranged in an array of direct tandem repeats. In an intra-species comparison, the sequences of the repeat units are relatively homogeneous; inter-species comparisons, however, show diversity except for a conserved "Goldberg-Hogness box", T-A-T-A-A-A-T-A. The size of a repeat unit and the number of repeats within a molecule can vary over a wide range, even in an intra-species comparison. Because of these wide inter-species variations observed, it is likely that the function of this region imposes few constraints on the sequence other than its high A + T content and possibly a Goldberg-Hogness box. The array of direct tandem repeats may have arisen from unequal recombination or crossover within this region. Adjacent to the non-palindromic region is a transcribed sequence which is highly conserved for all species and presumably represents a gene coding region.


Potential protein biomarkers for systemic lupus erythematosus determined by bioinformatics analysis.
Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disorder, and its pathogenesis in males and in cases without accompanying lupus nephritis (LN-) is not fully understood. In this study, we identified 90 (82 up- and 8 downregulated) differentially expressed genes (DEGs) common to female LN-, female LN+ and male LN+ using the GSE65391 and GSE49454 gene expression datasets from Gene Expression Omnibus database (GEO). The protein-protein interaction (PPI) network of 70 DEGs was constructed using STRING and cytoscape, and the Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the PPI network was significantly enriched in defense response to virus, cytosol, protein binding and measles. Sixteen hubgenes were identified from this PPI network, and Literature Mining Gene Networks molecular of GenCLiP 2.0 showed strong interaction between STAT1, DDX58 and IFIT1. Enrichment analysis of hubgenes in published literature showed the involvement of immune response and interferon-related genes in the pathogenesis of SLE. In addition, the transcription factors STAT1 & 2 and IRF6 & 9 had high Normalized Enrichment Score (NES). The 70 DEGs with PPI network and 16 hubgenes are potential biomarkers of SLE, and can help improve diagnosis and develop individualized therapies.


Organic and elemental carbon associated to PM10 and PM 2.5 at urban sites of northern Greece.
Organic carbon (OC) and elemental carbon (EC) concentrations, associated to PM10 and PM2.5 particle fractions, were concurrently determined during the warm and the cold months of the year (July-September 2011 and February-April 2012, respectively) at two urban sites in the city of Thessaloniki, northern Greece, an urban-traffic site (UT) and an urban-background site (UB). Concentrations at the UT site (11.3 ± 5.0 and 8.44 ± 4.08 14 μg m(-3) for OC10 and OC2.5 vs. 6.56 ± 2.14 and 5.29 ± 1.54 μg m(-3) for EC10 and EC2.5) were among the highest values reported for urban sites in European cities. Significantly lower concentrations were found at the UB site for both carbonaceous species, particularly for EC (6.62 ± 4.59 and 5.72 ± 4.36 μg m(-3) for OC10 and OC2.5 vs. 0.93 ± 0.61 and 0.69 ± 0.39 μg m(-3) for EC10 and EC2.5). Despite that, a negative UT-UB increment was frequently evidenced for OC2.5 and PM2.5 in the cold months possibly indicative of emissions from residential wood burning at the urban-background site. At both sites, cconcentrations of OC fractions were significantly higher in the cold months; on the contrary, EC fractions at the UT site were prominent in the warm season suggesting some influence from maritime emissions in the nearby harbor area. Secondary organic carbon, being estimated using the EC tracer method and seasonally minimum OC/EC ratios, was found to be an appreciable component of particle mass particularly in the cold season. The calculated secondary contributions to OC ranged between 35 and 59 % in the PM10 fraction, with relatively higher values in the PM2.5 fraction (39-61 %). The source origin of carbonaceous species was investigated by means of air parcel back trajectories, satellite fire maps, and concentration roses. A local origin was mainly concluded for OC and EC with limited possibility for long range transport of biomass (agricultural waste) burning aerosol.


SYTL2 promotes metastasis of prostate cancer cells by enhancing FSCN1-mediated pseudopodia formation and invasion.
Metastatic prostate cancer (mPCa) has a poor prognosis with limited treatment options. The high mobility of tumor cells is the key driving characteristic of metastasis. However, the mechanism is complex and far from clarified in PCa. Therefore, it is essential to explore the mechanism of metastasis and discover an intrinsic biomarker for mPCa. Transcriptome sequencing data and clinicopathologic features of PCa from multifarious public databases were used to identify novel metastatic genes in PCa. The PCa tissue cohort containing 102 formalin-fixed paraffin-embedded (FFPE) samples was used to evaluate the clinicopathologic features of synaptotagmin-like 2 (SYTL2) in PCa. The function of SYTL2 was investigated by migration and invasion assays and a 3D migration model in vitro and a popliteal lymph node metastasis model in vivo. We performed coimmunoprecipitation and protein stability assays to clarify the mechanism of SYTL2. We discovered a pseudopodia regulator, SYTL2, which correlated with a higher Gleason score, worse prognosis and higher risk of metastasis. Functional experiments revealed that SYTL2 promoted migration, invasion and lymph node metastasis by increasing pseudopodia formation in vitro and in vivo. Furthermore, SYTL2 induced pseudopodia formation by enhancing the stability of fascin actin-bundling protein 1 (FSCN1) by binding and inhibiting the proteasome degradation pathway. Targeting FSCN1 enabled rescue and reversal of the oncogenic effect of SYTL2. Overall, our study established an FSCN1-dependent mechanism by which SYTL2 regulates the mobility of PCa cells. We also found that the SYTL2-FSCN1-pseudopodia axis may serve as a pharmacological and novel target for treating mPCa.


Insertion of chromosome 11 in chromosome 4 resulting in a 5'MLL-3'AF4 fusion gene in a case of adult acute lymphoblastic leukemia.
We report on a 69-year-old woman with B-lineage acute lymphoblastic leukemia. Cytogenetic studies at diagnosis with R banding showed an insertion, ins(4;11)(q21;q13q23). Fluorescence in situ hybridization (FISH) with whole chromosome painting probes confirmed the insertion of chromosome 11 material into chromosome 4. FISH using the MLL probe showed the translocation of the 5' end of MLL into chromosome 4. Since the 5'MLL-3'AF4 fusion transcript was detected by a reverse transcriptase polymerase chain reaction, we concluded that the insertion of part of chromosome 11 split the AF4 gene in two, resulting in the presence of the 5'MLL-3'AF4 fusion gene on the der(4) instead of the der(11), as commonly observed. Our findings stress the value of combining banding cytogenetics with FISH and molecular techniques to better assess rearrangements in leukemia.


Outcome of intracoronary stenting after failed maximal medical therapy in patients with symptomatic myocardial bridge.
The aim of the present study was to study the outcome of coronary stenting in patients with symptomatic myocardial bridging refractory to standard medical therapy. Medical therapy constitutes the first line treatment for patients with symptomatic myocardial bridge, surgical correction may be offered to select patients with refractory symptoms. Limited data exists on the outcome of coronary stenting in this patient cohort. Twenty-nine patients with symptoms of angina, documented myocardial ischemia and angiographically documented myocardial bridge of the left anterior descending artery (defined as [mt]50% systolic compression) were identified retrospectively. Seventeen (59%) patients responded to maximized medical therapy (mean age 51 +/- 16 years, range 29-80, 7 female) and 12 (41%) patients received coronary stents following a failed attempt of minimum 3 months of maximal medical therapy (mean age 49 +/- 15 years, range 32-76 years, 5 females). Medical therapy responders were compared with intracoronary stent group for recurrent severe angina, target vessel revascularization (TVR), myocardial infarction (MI), and death at follow-up. Intracoronary stents were placed in all patients successfully. The incidence of recurrent severe angina and TVR were significantly greater in the stent group (P = 0.04) while MI and death in two groups were similar at mean follow-up of 15 +/- 3 months. Coronary stent placement for medically refractory symptomatic myocardial bridge failed to relieve severe angina and is associated with high clinical restenosis and hence should be avoided.


Prophylactic Negative-pressure Dressings Reduce Wound Complications and Resource Burden After Emergency Laparotomies.
Surgical site infection (SSI) and wound breakdown after emergency laparotomy are common. They incur significant patient morbidity and health care costs. Negative-pressure dressings (NPDs) applied over closed incisions may minimize wound complications. However, its utility in the emergency setting is unknown. Here, we examined whether prophylactic NPD reduces wound complications after emergency laparotomies. This is a retrospective review of consecutive emergency laparotomies undertaken at a university hospital from January 2018 to October 2019. Outcomes included the rate of SSI, wound breakdown, hospital-outreach service utilization, wound-related readmissions, and length of stay. Propensity score matched analysis was used to assess bias. A total of 227 emergency laparotomies were reviewed, 70 received NPD and 157 had conventional dressings (controls). SSI was identified in 33 (21.0%) patients from the control group and six (8.6%) from the NPD group (odds ratio 0.35, 95% confidence interval: 0.15-0.85, P = 0.022). Wound breakdown was observed in 21 (13.4%) patients from the control group and three (4.3%) from the NPD group (odds ratio 0.29, 95% confidence interval: 0.09-0.91, P = 0.040). The prophylactic benefit of NPD was most evident in clean-contaminated, contaminated, and dirty wounds. The NPD group had comparatively shorter postoperative stay, less outreach service utilization, and lower rates of wound-related readmissions. Multivariate analysis demonstrated that increasing age, body weight >75 kg, and wound contamination are independent predictors of wound complications, whereas NPD prevented SSI and wound breakdown. Prophylactic NPD significantly reduced wound complications after emergency laparotomy. This was associated with a substantial health resource saving. This study provides a strong rationale for randomized trials in this area.


Regional hypothermia protects against tourniquet neuropathy.
Ischaemic nerve injury has been suggested as the mechanism for post-tourniquet limb paralysis. As hypothermia has been shown to prolong ischaemia time in many tissues, we tested the hypothesis that cold protection would reduce tourniquet induced neural injury. In 16 male Wistar rats a tourniquet was applied to the right hind limb for 3 h. In eight rats both hind limbs were maintained at room temperature (27 degrees C) while the limb temperature was decreased to 4 degrees C by application of cold polyglycol gel packs in the remaining eight animals. Motor nerve conduction velocity (MNCV) and the amplitude of muscle motor units (AMMU) in response to nerve stimulation were measured at 0 h, 1 h and 1 week post-tourniquet release. Application of the tourniquet for 3 h abolished nerve conduction in all animals. Hypothermia resulted in a significant improvement in AMMU (5.8 +/- vs. 0.6 +/- 0.4 mv, p less than 0.05) at 1 h and in MNCV (26.6 +/- 1.6 vs. 8.1 +/- 4.4 ms-1, p less than 0.05) and AMMU (18.5 +/- 2.0 vs. 9.7 +/- 4.5 mv, p less than 0.05) at 1 week. Hypothermia without ischaemia also reduced MNCV (44.2 +/- 9.4 vs. 74.3 +/- 8.9 ms-1, p less than 0.05) and AMMU (28.1 +/- 2.8 vs. 36.3 +/- 8.4 mv, N.S.). These data indicate that hypothermia reduces normal MNCV and protects nerve function during tourniquet ischaemia. These results support the ischaemic hypothesis for post-tourniquet nerve injury. Cold protection may have clinical applications for surgical procedures performed with tourniquet ischaemia.


Effectiveness of a community health worker-led case management programme to improve outcomes for people with psychotic disorders in Thailand: a one-year prospective cohort study.
Intensive case management (ICM) programmes for psychotic patients are effective in improving outcomes, but often unfeasible in resource-poor settings, as they typically require extensive human resources and expertise. We developed and evaluated the effectiveness of a less intensive case management program (LICM), led by community health workers, on one-year social functioning and service use. A prospective cohort study was conducted on patients aged 18 and above residing in a hospital catchment area. Outcomes were compared between LICM (n = 64) and non-LICM participants (n = 485). A counterfactual framework approach was applied to assess causal effects of the LICM on outcomes. The programme effectiveness was analyzed by augmented-inverse probability of treatment weighting (AIPW) to estimate potential outcome mean (POM) and average treatment effect (ATE). Outcomes were employment status and use of emergency, inpatient and outpatient services. Analyses were stratified by the number of previous psychotic relapse (≤ 1, > 1) to assess heterogeneity of treatment effect on those in an early and later stages of psychotic illness. In the early-stage cohort, the likelihood of being employed at one year post-baseline was significantly greater in LICM participants than non-LICM participants (ATE 0.10, 95%CI 0.05-0.14, p < 0.001), whereas service use of all types, except outpatient, was not significantly different between the two groups. In the later-stage cohort, the likelihoods of employment between the two groups at post-baseline were similar (ATE -0.02, 95%CI -0.19-0.15, p = 0.826), whereas service use of all types was significantly higher in LICM participants. LICM in a setting where community mental services are scarce may benefit those at an early stage of psychotic illness, by leading to better social functioning and no higher use of unscheduled services at the end of the programme, possibly through their better prognosis and medication adherence. A more intensive case management model may be appropriate for those in a later stage of the illness.


An online pilates exercise program is effective on proprioception and core muscle endurance in a randomized controlled trial.
Proprioception is important for stability of body segments, postural control, and functionality. However, there are no studies in literature showing effects of online Pilates exercises that create proprioceptive inputs on vertebra on trunk proprioception. This study aims to reveal effect of online Pilates exercises conducted on trunk proprioception and core muscle endurance in healthy individuals. We included thirty-three healthy individuals between ages of 18 and 25 in study. Individuals were randomly divided into two groups. There were 17 individuals in Pilates group (PG), and 16 individuals in control group (CG). The PG was given online Pilates exercises by the physiotherapist in groups 3 days a week for 6 weeks, 1 h a day. There was no exercise program recommended for individuals in CG. We evaluated trunk proprioception with an inclinometer, core muscle endurance with three core endurance tests created by McGill, and prone bridge tests. All evaluations completed just before start of study and 2 days after 6-week training. Two groups had similar demographic characteristics, and there was no difference between baseline measurements (p > 0.05). While improvement observed in PG in trunk proprioception and all of core muscle endurance tests (p < 0.05), no statistically significant difference reported in CG (p > 0.05). We revealed that online Pilates exercises performed at mat level for 6 weeks in healthy individuals had positive effects on trunk proprioception and core muscle endurance with this study. Contribution of Pilates exercises to development of both muscular endurance and proprioceptive senses, even if performed at a distance, is important.


Imaging of colorectal adenocarcinoma using FT-IR microspectroscopy and cluster analysis.
In this paper, three different clustering algorithms were applied to assemble infrared (IR) spectral maps from IR microspectra of tissues. Using spectra from a colorectal adenocarcinoma section, we show how IR images can be assembled by agglomerative hierarchical (AH) clustering (Ward's technique), fuzzy C-means (FCM) clustering, and k-means (KM) clustering. We discuss practical problems of IR imaging on tissues such as the influence of spectral quality and data pretreatment on image quality. Furthermore, the applicability of cluster algorithms to the spatially resolved microspectroscopic data and the degree of correlation between distinct cluster images and histopathology are compared. The use of any of the clustering algorithms dramatically increased the information content of the IR images, as compared to univariate methods of IR imaging (functional group mapping). Among the cluster imaging methods, AH clustering (Ward's algorithm) proved to be the best method in terms of tissue structure differentiation.


Description and cost-analysis of emergency department attendances for hand and wrist injuries.
Injuries to the hand and wrist are estimated to account for between 10% and 30% of all ED presentations. The economic burden placed on the healthcare system can be extensive and rise sharply with increase in injury severity. This cost-analysis was performed with the aim of estimating the economic implications of ED attendances for hand and wrist injuries from the perspective of one Australian public health network. Data from two EDs were retrieved from the electronic billing records of one large health network across two financial year periods (2014-2015 and 2015-2016) using ICD-10 codes. All costs that resulted from the treatment of any acute hand or wrist injury across the 2 year period were calculated and are presented by age, sex, injury type and mechanism of injury. A total of 10 024 individuals presented to the two EDs in the 2 year period, accounting for approximately 5.4% of all presentations. The most common presentations were males (62.2%), people aged 25-34 years (26.9%) and lacerations (31.2%). The total cost in the 2 year study period was $3 959 535.38 ($1 923 852.38 in 2014-2015; $2 035 683.00 in 2015-2016). The mean cost per presentation was $383 (95% CI [$373, $393]) in 2014-2015 and $407 (95% CI [$394, $421]) in 2015-2016. Acute hand and wrist injuries contribute to a significant volume of ED presentations each year in one Australian public health network leading to significant expenditure and health resources. Further research into how to best utilise resources and reduce avoidable injuries should be priority areas to reduce the cost of these injuries to the healthcare system and society.


Pinocchio cells: morphologically atypical immunologically heterogeneous lymphocytes induced by treatment with interleukin 2.
We describe a novel cell type, the Pinocchio cell, that appears in the peripheral blood of all patients receiving treatment with interleukin 2, up to 20,000 cells/microliter. This cell is characterized by a prominent and granular proboscis with which it attaches to tumor cells and mediates tumor cell lysis. Pinocchio cells are immunologically heterogeneous and express antigens of both T and NK cells; Pinocchio cells are adherent in culture and are more cytolytic than non-adherent cells against NK-sensitive and resistant tumor cells. Incubation of normal whole human blood for 1 h induces Pinocchio morphology of mononuclear white blood cells.


Selective staining of human platelet glycoproteins using nitrocellulose transfer of electrophoresed proteins and peroxidase-conjugated lectins.
Platelet proteins (0.5-5 micrograms) were electrophoresed in a one-dimensional or an unreduced-reduced, two-dimensional sodium dodecyl sulfate gel system. The separated proteins were then transferred electrophoretically to nitrocellulose and reacted with peroxidase-conjugated lectins. Visualization of specific glycoproteins which bound the lectins was made by the chromogenic reaction catalyzed by peroxidase utilizing 3,3'-diaminobenzidine as the substrate. Wheat germ agglutinin specifically reacted with and allowed the visualization of glycoprotein Ib. Peanut agglutinin also specifically stained glycoprotein Ib after treatment of the nitrocellulose transferred proteins with neuraminidase. Ricinus communis agglutinin I stained thrombospondin, a 260 kDa protein, and factor VIII. Concanavalin A stained mainly glycoproteins IIb, III, IV, and V. Glycoproteins Ia, Ic, IIa, and other minor glycoproteins could be separated by unreduced-reduced, two-dimensional gel electrophoresis and were stained weakly with wheat germ agglutinin conjugates. These techniques were found to be reproducible as well as easily applied to the analysis and identification of platelet glycoproteins, particularly when dealing with a limited amount of platelets.


The lineage-specific, intrinsically disordered N-terminal extension of monothiol glutaredoxin 1 from trypanosomes contains a regulatory region.
Glutaredoxins (Grx) are small proteins conserved throughout all the kingdoms of life that are engaged in a wide variety of biological processes and share a common thioredoxin-fold. Among them, class II Grx are redox-inactive proteins involved in iron-sulfur (FeS) metabolism. They contain a single thiol group in their active site and use low molecular mass thiols such as glutathione as ligand for binding FeS-clusters. In this study, we investigated molecular aspects of 1CGrx1 from the pathogenic parasite Trypanosoma brucei brucei, a mitochondrial class II Grx that fulfills an indispensable role in vivo. Mitochondrial 1CGrx1 from trypanosomes differs from orthologues in several features including the presence of a parasite-specific N-terminal extension (NTE) whose role has yet to be elucidated. Previously we have solved the structure of a truncated form of 1CGrx1 containing only the conserved glutaredoxin domain but lacking the NTE. Our aim here is to investigate the effect of the NTE on the conformation of the protein. We therefore solved the NMR structure of the full-length protein, which reveals subtle but significant differences with the structure of the NTE-less form. By means of different experimental approaches, the NTE proved to be intrinsically disordered and not involved in the non-redox dependent protein dimerization, as previously suggested. Interestingly, the portion comprising residues 65-76 of the NTE modulates the conformational dynamics of the glutathione-binding pocket, which may play a role in iron-sulfur cluster assembly and delivery. Furthermore, we disclosed that the class II-strictly conserved loop that precedes the active site is critical for stabilizing the protein structure. So far, this represents the first communication of a Grx containing an intrinsically disordered region that defines a new protein subgroup within class II Grx.


Silencing of the nuclear RPS10 gene encoding mitochondrial ribosomal protein alters translation in arabidopsis mitochondria.
Hardly anything is known about translational control of plant mitochondrial gene expression. Here, we provide evidence for differential translation of mitochondrial transcripts in Arabidopsis thaliana. We found that silencing of the nuclear RPS10 gene encoding mitochondrial ribosomal protein S10 disturbs the ratio between the small and large subunits of mitoribosomes, with an excess of the latter. Moreover, a portion of the small subunits are incomplete, lacking at least the S10 protein. rps10 cells also have an increased mitochondrial DNA copy number per cell, causing an upregulation of all mitochondrial transcripts. Mitochondrial translation is also altered so that it largely overrides the hyperaccumulation of transcripts, and as a consequence, only ribosomal proteins are oversynthesized, whereas oxidative phosphorylation subunits are downregulated. Expression of nuclear-encoded components of mitoribosomes and oxidative phosphorylation system (OXPHOS) complexes seems to be less affected. The ultimate coordination of expression of the nuclear and mitochondrial genomes occurs at the complex assembly level. These findings indicate that mitoribosomes can regulate gene expression by varying the efficiency of translation of mRNAs for OXPHOS and ribosomal proteins.


Ex Vivo Expansion and Drug Sensitivity Profiling of Circulating Tumor Cells from Patients with Small Cell Lung Cancer.
Small cell lung cancer (SCLC) represents one of the most aggressive malignancies among cancer types. Not only tumor sample availability is limited, but also the ability for tumor cells to rapidly acquire drug resistance are the rate-limiting bottlenecks for overall survival in current clinical settings. A liquid biopsy capable of capturing and enriching circulating tumor cells (CTCs), together with the possibility of drug screening, is a promising solution. Here, we illustrate the development of a highly efficient ex vivo CTC expansion system based on binary colloidal crystals substrate. Clinical samples were enrolled from 22 patients with SCLC in the study. The CTCs were enriched and expanded from the collected peripheral blood samples. Expanded cells were analyzed for protein expression and observed for drug sensitivity with the use of immunofluorescence and ATP titer evaluation, respectively. Successful CTC spheroid proliferation was established after 4 weeks within 82% of all the collected peripheral blood samples from enrolled patients. Upon immunofluorescence analysis, the enriched cells showed positive markers for EpCAM, TTF-1, synaptophysin and negative for CD45. Additionally, the expanded CTCs demonstrated marked heterogeneity in the expression of E-cadherin and N-cadherin. In a preliminary case series, the drug sensitivity of patient-derived CTC to cisplatin and etoposide was studied to see the correlation with the corresponding therapeutic outcome. In conclusion, our study demonstrates that it is possible to efficiently expand CTCs from SCLC within a clinically relevant time frame; the biomarker information generated from enriched CTCs can assist the selection of effective drugs and improve disease outcome.


Intraoperative fluorescence-guided resection of high-grade malignant gliomas using 5-aminolevulinic acid-induced porphyrins: a systematic review and meta-analysis of prospective studies.
We performed a systematic review and meta-analysis to address the (added) value of intraoperative 5-aminolevulinic acid (5-ALA)-guided resection of high-grade malignant gliomas compared with conventional neuronavigation-guided resection, with respect to diagnostic accuracy, extent of tumor resection, safety, and survival. An electronic database search of Medline, Embase, and the Cochrane Library was undertaken. The review process followed the guidelines of the Cochrane Collaboration. 10 studies matched all selection criteria, and were thus used for qualitative synthesis. 5-ALA-guided resection demonstrated an overall sensitivity of 0.87 (95% confidence interval [CI], 0.81-0.92), specificity of 0.89 (95% CI, 0.79-0.94), positive likelihood ratio (LR) of 7.62 (95% CI, 3.87-15.01), negative LR of 0.14 (95% CI, 0.09-0.23), and diagnostic odds ratio (OR) of 53.06 (95% CI, 18.70-150.51). Summary receiver operating characteristic curves (SROC) showed an area under curve (AUC) of 94%. Contrast-enhancing tumor was completely resected in patients assigned 5-ALA as compared with patients assigned white light. Patients in the 5-ALA group had higher 6-month progression free survival and overall survival than those in the white light group. Based on available literature, there is level 2 evidence that 5-ALA-guided surgery is more effective than conventional neuronavigation-guided surgery in increasing diagnostic accuracy and extent of tumor resection, enhancing quality of life, or prolonging survival in patients with high-grade malignant gliomas.


Immunological aspects of acute ureteral obstruction: immune cell infiltrate in the kidney.
Kidneys from rats subjected to bilateral ureteral obstruction (BUO), unilateral ureteral obstruction (UUO) and UUO with subsequent release were analyzed for leukocyte infiltration. A time-dependent influx of leukocytes, predominantly macrophages and suppressor T lymphocytes, occurred in both the cortex and medulla following obstruction, and disappeared with release of the obstruction. Glomerular macrophages declined following obstruction but increased to levels above control following release. The influx of leukocytes following obstruction was paralleled by an increase in thromboxane B2 excretion by the kidney and coincided with a decrease in glomerular filtration rate (GFR). This would suggest that an influx of immune cells is a prominent feature of the acute renal response to ureteral obstruction. These cells may modulate some of the post-obstructive alterations in renal function via the production of vasoactive substances, such as thromboxane A2.


Utility of B-natriuretic peptide in detecting diastolic dysfunction: comparison with Doppler velocity recordings.
Although Doppler echocardiography has been used to identify abnormal left ventricular (LV) diastolic filling dynamics, inherent limitations suggest the need for additional measures of diastolic dysfunction. Because data suggest that B-natriuretic peptide (BNP) partially reflects ventricular pressure, we hypothesized that BNP levels could predict diastolic abnormalities in patients with normal systolic function. We studied 294 patients referred for echocardiography to evaluate ventricular function. Patients with abnormal systolic function were excluded. Cardiologists making the assessment of LV function were blinded to BNP levels. Patients were classified as normal, impaired relaxation, pseudonormal, and restrictivelike filling patterns. Patients diagnosed with evidence of abnormal LV diastolic function (n=119) had a mean BNP concentration of 286 +/- 31 pg/mL; those in the normal LV group (n=175) had a mean BNP concentration of 33 +/- 3 pg/mL. Patients with restrictive like filling patterns on echocardiography had the highest BNP levels (408 +/- 66 pg/mL), and patients with symptoms had higher BNP levels in all diastolic filling patterns. The area under the receiver-operating characteristic curve for BNP to detect any diastolic dysfunction was 0.92 (95% CI, 0.87 to 0.95; P<0.001). A BNP value of 62 pg/mL had a sensitivity of 85%, a specificity of 83%, and an accuracy of 84% for detecting diastolic dysfunction. A rapid assay for BNP can reliably detect the presence of diastolic abnormalities on echocardiography. In patients with normal systolic function, elevated BNP levels and diastolic filling abnormalities might help to reinforce the diagnosis diastolic dysfunction.


Triterpene saponosides from Lysimachia ciliata differentially attenuate invasive potential of prostate cancer cells.
Neither androgen ablation nor chemotherapeutic agents are effective in reducing the risk of prostate cancer progression. On the other hand, multifaceted effects of phytochemicals, such as triterpene saponins, on cancer cells have been suggested. A promising safety and tolerability profile indicate their possible application in the treatment of advanced prostate cancers. We analyzed the specificity, selectivity and versatility of desglucoanagalloside B effects on human prostate cancer cells derived from prostate cancer metastases to brain (DU-145 cells) and bone (PC-3 cells). Prominent growth arrest and apoptotic response of both cell types was observed in the presence of sub-micromolar desglucoanagalloside B concentrations. This was accompanied by cytochrome c release and caspase 3/7 activation. A relatively low cytostatic and pro-apoptotic response of cancer cells to a desglucoanagalloside B analog, anagallosaponin IV, illustrated the specificity of the effects of desglucoanagalloside B, whereas the low sensitivity of normal prostate PNT2 cells to desglucoanagalloside B showed the selectivity of its action. Inhibition of cancer cell motility was observed in the presence of both saponins, however only desglucoanagalloside B attenuated cancer cell invasive potential, predominantly through an effect on cell elastic properties. These data demonstrate the versatility of its effects on prostate cancer cells. In contrast to PNT2 cells, cancer cells tested in this study were relatively resistant to mitoxantrone. The multifaceted action of desglucoanagalloside B on basic cellular traits, crucial for prostate cancer progression, opens perspectives for elaboration of combined palliative therapies and new prostate cancer prophylaxis regimens.


The OH-initiated oxidation of 1,3-butadiene in the presence of O2 and NO: a photolytic route to study isomeric selective reactivity.
We report the study of the isomeric selective OH-initiated oxidation of 1,3-butadiene in the presence of O2 and NO using the LP/LIF technique. The photolysis of monodeuterated 1-iodo-3-buten-2-ol provides only one of the possible OD-butadiene adducts, the minor addition channel product, simplifying the oxidation mechanism. We find, based on analysis of OD time-dependent traces that prompt rearrangement of initial beta-hydroxyalkyl radicals to alpha-hydroxyalkyl radicals occurs in agreement with RRKM/ME theoretical predictions. We report a rate constant of (3.3+/-1.0) x 10(-11) cm3 molecules(-1) s(-1) for deuterium abstraction from the alpha-hydroxyalkyl radical at 298 +/-2 K. Our approach demonstrates the feasibility of isomeric selective kinetic studies of the OH-initiated oxidation of unsaturated hydrocarbons.


Association between vaccination and preventive routines on COVID-19-related mortality in nursing home facilities: a population-based systematic retrospective chart review.
Older and frail individuals are at high risk of dying from COVID-19, and residents in nursing homes (NHs) are overrepresented in death rates. We explored four different periods during the COVID-19 pandemic to analyze the effects of improved preventive routines and vaccinations, respectively, on mortality in NHs. We undertook a population-based systematic retrospective chart review comprising 136 NH facilities in southeast Sweden. All residents, among these facilities, who died within 30 days after a laboratory-verified COVID-19 diagnosis during four separate 92-day periods representing early pandemic (second quarter 2020), middle of the pandemic (fourth quarter 2020), early post-vaccination phase (first quarter 2021), and the following post-vaccination phase (second quarter 2021). Mortality together with electronic chart data on demographic variables, comorbidity, frailty, and cause of death was collected. The number of deaths during the four periods was 104, 120, 34 and 4, respectively, with a significant reduction in the two post-vaccination periods (P < 0.001). COVID-19 was assessed as the dominant cause of death in 20 (19%), 19 (16%), 4 (12%) and 1 (3%) residents in each period (P < 0.01). The respective median age in the four studied periods varied between 87and 89 years, and three or more diagnoses besides COVID-19 were present in 70-90% of the respective periods' study population. Considerable or severe frailty was found in all residents. Vaccination against COVID-19 seems associated with a reduced number of deaths in NHs. We could not demonstrate an effect on mortality merely from the protective routines that were undertaken.


Physical training and fasting erythrocyte activities of free radical scavenging enzyme systems in sedentary men.
Effects of 10 weeks of physical training on free radical scavenging enzyme systems in erythrocytes were investigated in 7 sedentary healthy male students. The training consisted of running over 5 km, 6 times/week. Their maximum oxygen uptake and 12 min walk-run performance increased significantly after training. Of the antioxidant enzyme systems examined in the erythrocytes, both catalase activity and concentration and total glutathione reductase (GR) activity also showed significant increases following the training. The erythrocyte GR activity coefficient also increased significantly. These results suggest that chronic aerobic exercise increases riboflavin requirements and has some positive effects on antioxidative processes.


New concepts in the chromatography of peptides and proteins.
Although chromatography using a variety of novel bed configurations (e.g. fluidized beds, expanded beds, simulated moving beds, annular rotating beds, etc.) has been of recent interest, the majority of practical applications of analytical and preparative chromatography employ a stationary adsorbent bed into which a feed slug is charged periodically, similar to the technique first described by Mikhail Tswett over 100 years ago. However, new concepts in both the practice and theory of fixed-bed chromatography are continuing to expand the available range of applications for separating peptides and proteins.


Synthesis and characterization of the oxidized dGTP lesions spiroiminodihydantoin-2'-deoxynucleoside-5'- triphosphate and guanidinohydantoin-2'-deoxynucleoside-5'- triphosphate.
Two convenient synthetic routes to the oxidized guanosine triphosphate lesions spiroiminodihydantoin-2'-deoxynucleoside-5'-triphosphate (dSpTP) and guanidinohydantoin-2'-deoxynucleoside-5'-triphosphate (dGhTP) are reported. Both two-electron oxidation of 2'-deoxy-7,8-dihydro-8-oxoguanosine-5'-triphosphate (dOGTP) using SO4*- generated photolytically from K2S2O8 or four-electron oxidation of 2'-deoxyguanosine-5'-triphosphate (dGTP) from singlet oxygen provide either dSpTP or dGhTP at pH 8.0 or 4.4, respectively. Highly purified triphosphates are obtained by ion pair reversed-phase HPLC.


Altered activation pattern in synergistic ankle plantarflexor muscles in a reduced-gravity environment.
We investigated relative activations in the synergistic ankle plantarflexor muscles, the soleus (SOL) and medial gastrocnemius (MG), while subjects were walking in water. Ten healthy volunteers walked in water under three different load- and four different speed-conditions. The depth of water was to the axilla of each subject, which reduced about 80% of his or her body weight. The EMG activities of the SOL and MG were recorded with surface electrodes, and ground reaction forces (GRF) were measured by using a water-resistant force platform. Relations of EMG activity levels between the two muscles during the stance phase were analysed with respect to the load and walking speed. The results revealed that the SOL EMG activity was more dependent on the load than the walking speed in water, while the MG EMG activity was more dependent on the speed. Such dissociated activation of the synergistic ankle plantarflexors might be relevant to the different roles required of these muscles in functional motor tasks.


Lean mass mediates the relation between temporal summation of pain and sex in young healthy adults.
Previous studies have shown that women experience greater temporal summation (TS) of pain than men using a repetitive thermal stimulus. These studies, however, did not individualize the thermal stimulus to each subject's thermal pain sensitivity. The aim of this study was to investigate sex differences in TS using an individualized protocol and potential mediators that have been shown to influence TS including physical activity and body composition. Fifty young healthy men and women (21 men) participated in the study. Subjects completed TS testing on the right forearm using a repetitive thermal stimulus at a temperature that the subject reported 6/10 pain. Other testing included body composition (lunar iDXA), activity monitoring (Actigraph), and Pain Catastrophizing Scale (PCS). Women reported greater TS than men (p = 0.019), and TS was correlated with right arm lean mass (r = - 0.36, p = 0.01) and magnification subscale of PCS (r = - 0.32, p = 0.03). Mediation analysis showed a complete mediation for the relation between sex and TS by right arm lean mass (indirect effect = 2.33, 95% BCa CI [0.42, 4.58]) after controlling for the temperature, the magnification subscale of PCS, and the average time spent in moderate to vigorous physical activity. The results of this study suggest that lean mass is a contributing factor to the sex differences in TS. Future studies should investigate whether interventions that increase lean mass have a positive effect on TS.


Construction of Recombinant Lactococcus lactis Strain Expressing VP1 Fusion Protein of Duck Hepatitis A Virus Type 1 and Evaluation of Its Immune Effect.
With the continuous development of duck farming and the increasing breeding density, the incidence of duck hepatitis A virus type 1 (DHAV-1) has been on the rise, seriously endangering the development of duck farming. To reduce the use of antibiotics in duck breeding, susceptibility risks and mortality, and avoid virulence recovery and immune failure risk, this study aims to develop a new type of mucosal immune probiotics and make full use of molecular biology techniques, on the level of genetic engineering, to modify Lactococcus lactis (L. lactis). In this study, a secretory recombinant L. lactis named MG1363-VP1 with an enhanced Green Fluorescent Protein (eGFP) and translation enhancer T7g10L was constructed, which could express the VP1-eGFP fusion protein of DHAV-1. The animal experiment in ducklings was performed to detect the immune response and protection effect of oral microecologics by recombinant L. lactis. The results showed that oral L. lactis MG1363-VP1 significantly induced the body's humoral immune system and mucosal immune system to produce specific anti-VP1 IgG antibodies and mucosal secretory immunoglobulin A (sIgA) for DHAV-1 in ducklings, and cytokines including interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10), and interferon gamma (IFN-γ). The mortality rate was monitored simultaneously by the natural infestation in the process of production and breeding; notably, the ducklings vaccinated with L. lactis MG1363-VP1 were effectively protected against the nature infection of DHAV-1. The recombinant L. lactis MG1363-VP1 constructed in this study provides a new means of preventing and controlling DHAV-1 infection in the future.


Last performance with VIAGRA: post-mortem identification of sildenafil and its metabolites in biological specimens including hair sample.
A 43-year-old man was found dead in a hotel room during a sexual relation with a colleague.He was treated both for cardiovascular disease and for erectile dysfunction with VIAGRA. A pillbox was found in the room with several tablets of verapamil (Isoptine), trimetazidine (Vastarel), yohimbine and bromazepam (Lexomil). A box of VIAGRA 25mg was found in his raincoat and two tablets were missing. His wife declared during the investigation that he was also treated by trinitrine. Autopsy revealed severe coronary artery sclerosis as well as signs of previous myocardial infarctions. Blood, urine, bile, gastric content and hair and representative tissues for histology were collected for toxicological analysis. Sildenafil and yohimbine were screened with liquid chromatography/mass spectrometry (LC/MS) and trinitrine with headspace injection (HS)/GC/MS. Verapamil and trimetazidine were identified and quantified with LC/diode array detection (DAD). Sildenafil was identified in blood, urine, bile and gastric content at 105, 246, 1206 and 754ng/ml, respectively. Hair concentration was 177pg/mg. The desmethyl metabolite was quantified in urine at 143ng/ml. Blood concentrations of verapamil and trimetazidine were measured at 659 and 2133ng/ml, respectively and were above therapeutic ranges. Trinitrine and yohimbine were not identified. These results confirm the absorption of sildenafil, verapamil and trimetazidine before the death and hair analysis indicates the chronic use of sildenafil. To the author's knowledge, this is the first report of a fatal sildenafil-verapamil association, probably by hypotension and cardiac dysrhythmia.


Effect of pH and moderate hyperthermia on doxorubicin, epirubicin and aclacinomycin A cytotoxicity for Chinese hamster ovary cells.
The influence of extracellular pH on the cytotoxicity of the anthracyclines doxorubicin, epirubicin, and aclacinomycin A was examined at 37 degrees C and 41 degrees C in tissue culture. Chinese hamster ovary (CHO) cells were exposed for a total of 24 h to anthracyclines at doses ranging between 0.12 and 0.69 nmol/ml at pH 7.4, 6.7, and 6.4 and at 37 degrees C. Temperature elevation to 41 degrees C was carried out for 3 h after the initiation of the drug treatment. Doxorubicin and epirubicin were about equally cytotoxic in the pH range examined at both temperatures. Aclacinomycin A demonstrated a higher cytotoxicity at pH 7.4 and 37 degrees C only at low doses. At low pH, however, aclacinomycin A was increasingly more effective with increasing dose as compared with doxorubicin and epirubicin. At 41 degrees C and at higher doses aclacinomycin A was even less cytotoxic than doxorubicin or epirubicin. Doxorubicin and epirubicin were less effective at lower pH. However, aclacinomycin A at doses of greater than 0.25 nmol/ml was more cytotoxic at low pH. Moderate hyperthermia did not increase the cytotoxicity of the three drugs at low pH, except for aclacinomycin A at doses of less than 0.25 nmol/ml. At pH 7.4, aclacinomycin A was even less effective at the elevated temperature. At doses of greater than 0.25 nmol/ml, moderate hyperthermia decreased the cytotoxicity of aclacinomycin A at low pH.


Profiling cholinesterase adduction: a high-throughput prioritization method for organophosphate exposure samples.
A high-throughput prioritization method was developed for use with a validated confirmatory method detecting organophosphorus nerve agent exposure by immunomagnetic separation high-performance liquid chromatography tandem mass spectrometry. A ballistic gradient was incorporated into this analytical method to profile unadducted butyrylcholinesterase (BChE) in clinical samples. With Zhang et al.'s Z' factor of 0.88 ± 0.01 (SD) of control analytes and Z factor of 0.25 ± 0.06 (SD) of serum samples, the assay is rated an "excellent assay" for the synthetic peptide controls used and a "double assay" when used to prioritize clinical samples. Hits, defined as samples containing BChE Ser-198 adducts or no BChE present, were analyzed in a confirmatory method for identification and quantitation of the BChE adduct, if present. The ability to prioritize samples by highest exposure for confirmatory analysis is of particular importance in an exposure to cholinesterase inhibitors such as organophosphorus nerve agents, in which a large number of clinical samples may be collected. In an initial blind screen, 67 of 70 samples were accurately identified, giving an assay accuracy of 96%, and it yielded no false-negatives. The method is the first to provide a high-throughput prioritization assay for profiling adduction of Ser-198 BChE in clinical samples.


Embryo quality is the main factor affecting cumulative live birth rate after elective single embryo transfer in fresh stimulation cycles.
The study was aimed to evaluate which factors affect the cumulative live birth rate after elective single embryo transfer in women younger than 36 years. Additionally, number of children in women with more than one delivery per ovum pick-up after fresh elective single embryo transfer and subsequent frozen embryo transfers was assessed. Retrospective cohort study analysing data of a university hospital's infertility clinic in 2001-2010. A total of 739 IVF/ICSI cycles with elective single embryo transfer were included. Analyses were made per ovum pick-up including fresh and subsequent frozen embryo transfers. Factors affecting cumulative live birth rates were examined in uni- and multivariate analyses. A secondary endpoint was the number of children born after all treatments. In the fresh cycles, the live birth rate was 29.2% and the cumulative live birth rate was 51.3%, with a twin rate of 3.4%. In the multivariate analysis, having two (odds ratio (OR) 1.73; 95% confidence interval (CI) 1.12-2.67) or ≥3 top embryos (OR 2.66; 95% CI 1.79-3.95) was associated with higher odds for live birth after fresh and frozen embryo cycles. Age, body mass index, duration of infertility, diagnosis or total gonadotropin dose were not associated with the cumulative live birth rate. In cycles with one top embryo, the cumulative live birth rate was 40.2%, whereas it was 64.1% in those with at least three top embryos. Of women who had a live birth in the fresh cycle, 20.4% had more than one child after all frozen embryo transfers. Among women with three or more top embryos after ovum pick-up, 16.1% gave birth to more than one child. The cumulative live birth rate in this age group varies from 40% to 64% and is dependent on the quality of embryos. Women with three or more top embryos have good chance of having more than one child per ovum pick-up without elevated risk of multiple pregnancies.


[Turner's syndrome and autoimmune dysthyroidism].
The authors report a case of Turner's syndrome associated with Hashimoto's thyroiditis and present the various physiopathological hypotheses currently offered to explain the frequent association of an autoimmune pathology and a gonadal dysgenesis with an absence of X chromosome which is held responsible for autoimmunity. Is the chromosomal abnormality a factor predisposing to autoimmunity or does it result from a disturbance of the immune mechanism? Environmental factors, such as bacteria, viruses or some pollutants might be responsible for both the immune disorders and the chromosomal abnormality. The association of the two types of disease may also be accidental. In the present state of our knowledge, there is no answer to these questions.


Photoredox-Catalyzed Synthesis of α-Amino Acid Amides by Imine Carbamoylation.
An operationally simple protocol for the photocatalytic carbamoylation of imines is reported. Easily available, bench-stable 4-amido Hantzsch ester derivatives serve as precursors to carbamoyl radicals that undergo rapid addition to N-aryl imines. The reaction proceeds under blue light irradiation in the presence of the photocatalyst 3DPAFIPN and Brønsted/Lewis acid additives. Mechanistic studies indicated a photoredox mechanism that involves carbamoyl radicals.


Asymptotic Symmetries in the BV-BFV Formalism.
We show how to derive asymptotic charges for field theories on manifolds with "asymptotic" boundary, using the BV-BFV formalism. We also prove that the conservation of said charges follows naturally from the vanishing of the BFV boundary action, and show how this construction generalises Noether's procedure. Using the BV-BFV viewpoint, we resolve the controversy present in the literature, regarding the status of large gauge transformation as symmetries of the asymptotic structure. We show that even though the symplectic structure at the asymptotic boundary is not preserved under these transformations, the failure is governed by the corner data, in agreement with the BV-BFV philosophy. We analyse in detail the case of electrodynamics and the interacting scalar field, for which we present a new type of duality to a sourced two-form model.


The effect of weight loss and inappropriate plasma dexamethasone levels on the DST.
Recent studies from different research groups have raised fundamental questions about the postulated specificity of the dexamethasone suppression test (DST) for endogenous depression. Findings in 116 psychiatric inpatients and 24 semi-starved healthy volunteers underline the importance of weight loss as a factor affecting DST results. A study of 160 DSTs in 93 psychiatric inpatients further revealed a significant negative correlation of plasma cortisol and plasma dexamethasone levels 10 hours after oral administration of 1 mg of dexamethasone. These results suggest a decisive effect of the pharmacokinetics of dexamethasone, at least on the 1-mg DST.


Role of the neutral endopeptidase 24.11 in the conversion of big endothelins in guinea-pig lung parenchyma.
1. We have studied the conversion of big endothelin-1 (big ET-1), big endothelin-2 (big ET-2) and big endothelin-3 (big ET-3) and characterized the enzyme involved in the conversion of the three peptides in guinea-pig lung parenchyma (GPLP). 2. Endothelin-1 (ET-1), endothelin-2 (ET-2) and endothelin-3 (ET-3) (10 nM to 100 nM) caused similar concentration-dependent contractions of strips of GPLP. 3. Big ET-1 and big ET-2 also elicited concentration-dependent contractions of GPLP strips. In contrast, big ET-3, up to a concentration of 100 nM, failed to induce a contraction of the GPLP. 4. Incubation of strips of GPLP with the dual endothelin converting enzyme (ECE) and neutral endopeptidase (NEP) inhibitor, phosphoramidon (10 microM), as well as two other NEP inhibitors thiorphan (10 microM) or SQ 28,603 (10 microM) decreased by 43% (P < 0.05), 42% (P < 0.05) and 40% (P < 0.05) the contractions induced by 30 nM of big ET-1 respectively. Captopril (10 microM), an angiotensin-converting enzyme inhibitor, had no effect on the contractions induced by big ET-1. 5. The incubation of strips of GPLP with phosphoramidon (10 microM), thiorphan (10 microM) or SQ 28,603 (10 microM) also decreased by 74% (P < 0.05), 34% and 50% (P < 0.05) the contractions induced by 30 nM big ET-2 respectively. As for the contractions induced by big ET-1, captopril (10 microM) had no effect on the concentration-dependent contractions induced by big ET-2. 6. Phosphoramidon (10 microM), thiorphan (10 microM) and SQ 28,603 (10 microM) significantly potentiated the contractions of strips of GPLP induced by both ET-1 (30 nM) and ET-3 (30 nM). However, the enzymatic inhibitors did not significantly affect the contractions induced by ET-2 (30 nM) in this tissue. 7. These results suggest that the effects of big ET-1 and big ET-2 result from the conversion to ET-1 and ET-2 by at least one enzyme sensitive to phosphoramidon, thiorphan and SQ 28,603. This enzyme corresponds possibly to EC 126.96.36.199 (NEP 24.11) and could also be responsible for the degradation of ETs in the GPLP.


Age differences in discrimination of simulated single-formant frequency transitions.
We studied auditory discrimination of simulated single-formant frequency transitions that resembled portions of certain speech consonants. Significant age differences in transition discrimination occurred; both children and older adults required larger acoustic differences between transitions for discrimination than did teenagers/young adults. Longer transitions were more easily discriminated than shorter transitions by all listeners, and there were no differences between discriminations of rising and falling transitions. Teens/young adults and older adults, but not children, required larger frequency differences to discriminate frequency transitions followed by a steady-state sound than for transitions alone. There were also age differences in discrimination of steady-state sounds. These developmental-perceptual differences may help explain why children and older adults who have good pure-tone sensitivity may experience difficulty in understanding speech.


Targetable Tetrazine-Based Dynamic Nuclear Polarization Agents for Biological Systems.
Dynamic nuclear polarization (DNP) has shown great promise as a tool to enhance the nuclear magnetic resonance signals of proteins in the cellular environment. As sensitivity increases, the ability to select and efficiently polarize a specific macromolecule over the cellular background has become desirable. Herein, we address this need and present a tetrazine-based DNP agent that can be targeted selectively to proteins containing the unnatural amino acid (UAA) norbornene-lysine. This UAA can be introduced efficiently into the cellular milieu by genetic means. Our approach is bio-orthogonal and easily adaptable to any protein of interest. We illustrate the scope of our methodology and investigate the DNP transfer mechanisms in several biological systems. Our results shed light on the complex polarization-transfer pathways in targeted DNP and ultimately pave the way to selective DNP-enhanced NMR spectroscopy in both bacterial and mammalian cells.


Comparison of the detection of periodontal pathogens in bacteraemia after tooth brushing by culture and molecular techniques.
The prevalence and amounts of periodontal pathogens detected in bacteraemia samples after tooth brushing-induced by means of four diagnostic technique, three based on culture and one in a molecular-based technique, have been compared in this study. Blood samples were collected from thirty-six subjects with different periodontal status (17 were healthy, 10 with gingivitis and 9 with periodontitis) at baseline and 2 minutes after tooth brushing. Each sample was analyzed by three culture-based methods [direct anaerobic culturing (DAC), hemo-culture (BACTEC), and lysis-centrifugation (LC)] and one molecular-based technique [quantitative polymerase chain reaction (qPCR)]. With culture any bacterial isolate was detected and quantified, while with qPCR only Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were detected and quantified. Descriptive analyses, ANOVA and Chi-squared tests, were performed. Neither BACTEC nor qPCR detected any type of bacteria in the blood samples. Only LC (2.7%) and DAC (8.3%) detected bacteraemia, although not in the same patients. Fusobacterium nucleatum was the most frequently detected bacterial species. The disparity in the results when the same samples were analyzed with four different microbiological detection methods highlights the need for a proper validation of the methodology to detect periodontal pathogens in bacteraemia samples, mainly when the presence of periodontal pathogens in blood samples after tooth brushing was very seldom.


Evaluation of stress induced in peri-implant bone tissue by misfit in multi-implant prosthesis.
The objective of the present work was to use numerical analysis to evaluate the relevance of stress states induced in peri-implant bone tissue by a misfit in a dental fixed prosthesis. Misfits in both mesial-distal and lingual-labial directions were considered to investigate a realistic configuration of the problem. A finite element model of a portion of a mandible with two implants connected by a gold alloy bar was defined on the basis of the morphometric data of a partial edentulous patient. A specific procedure was developed to represent the bar-implant coupling in the case of a misalignment between the implant and central axes of the anchoring site to the bar. Loading conditions related to occlusal forces were also considered. The numerical analysis of the implant-bridge misfit showed significant stress effects on the peri-implant bone tissue. For the specific prosthetic configuration considered, the maximum compressive stress was in the range of 40-60 MPa, depending on the misfit considered. The stress level was largely affected by the axial and bending stiffness characteristics of bar-implant compound. Stress analysis using numerical methods made it possible to estimate stress states with high accuracy in terms of intensity and location. For the case considered, stress induced by misfit was comparable with that related to occlusal forces. Therefore, a possible bar-implant misfit should be carefully considered to ensure the reliability of the prosthetic system.


Cranial manifestations of aspergillosis.
Five patients with differing forms and severity of cranial aspergillosis illustrate the range of neuroradiologic manifestations as evaluated with angiography, computed tomography and magnetic resonance imaging. Together with the physical findings, these subjects illustrate the dire clinical situation requiring immediate recognition in order to attempt to control this devastating, treatment resistant pathologic fungal infection.


Surgical implications of the potential new tubal pathway for ovarian carcinogenesis.
Since 2001, many studies by different investigators have demonstrated that the fallopian tube might be at the origin of most high-grade ovarian and peritoneal serous carcinomas. Simple changes in surgical practice (ie, prophylactic bilateral salpingectomy instead of salpingo-oophorectomy) could have significant implications for death from ovarian cancer and, on the other hand, for the morbidity caused by ovariectomy (surgical menopause). In this review, we describe the new tubal carcinogenic sequence, the advantages and disadvantages of exclusive use of salpingectomy in the general population, and in cases of hereditary predisposition to ovarian cancer such as for carriers of BRCA mutation.