Intravenous gamma globulin therapy in systemic juvenile rheumatoid arthritis.
Intravenous (IV) gamma globulin has been successfully used as replacement therapy for antibody-deficient patients and, more recently, in the treatment of autoimmune diseases such as idiopathic thrombocytopenic purpura, myasthenia gravis, and Kawasaki disease. In view of the successful treatment of these diseases, we initiated a pilot study of the effect of IV gamma globulin in systemic juvenile rheumatoid arthritis (JRA). Eight patients with active systemic JRA that was unresponsive to first-line agents, second-line agents, and/or corticosteroids received this therapy monthly for 6 months. Outcome measures included changes in articular and extraarticular features, steroid dosage, and laboratory parameters. Following IV gamma globulin therapy, there was significant improvement in arthritis and/or morning stiffness in 5 of 8 patients, while extraarticular features significantly improved in 7 of 8 patients. At study entry, 6 of 8 patients were receiving prednisone; at study end, prednisone was discontinued in 3 patients and decreased by more than 50% in the other 3. Overall, there was an 80% reduction in the prednisone dosage. Initially, all patients had anemia, low levels of serum albumin, and an elevated erythrocyte sedimentation rate, while a thrombocytosis was seen in 7 of 8 patients. Serum IgG was initially elevated in 6 patients. IV gamma globulin therapy resulted in a significant increase in hemoglobin and albumin levels and a significant decrease in the mean serum IgG level, platelet count, and erythrocyte sedimentation rate. In only 1 patient did IV gamma globulin fail to significantly improve the clinical or laboratory features of the disease. We suggest that this therapy may be beneficial in the treatment of systemic JRA.


Isolation of a Miller-Dieker lissencephaly gene containing G protein beta-subunit-like repeats.
Lissencephaly (agyria-pachygyria) is a human brain malformation manifested by a smooth cerebral surface and abnormal neuronal migration. Identification of the gene(s) involved in this disorder would facilitate molecular dissection of normal events in brain development. Type 1 lissencephaly occurs either as an isolated abnormality or in association with dysmorphic facial appearance in patients with Miller-Dieker syndrome. About 15% of patients with isolated lissencephaly and more than 90% of patients with Miller-Dieker syndrome have microdeletions in a critical 350-kilobase region in chromosome 17p13.3 (ref. 6). These deletions are hemizygous, so haplo-insufficiency for a gene in this interval is implicated. Here we report the cloning of a gene (LIS-1, lissencephaly-1) in 17p13.3 that is deleted in Miller-Dieker patients. Non-overlapping deletions involving either the 5' or 3' end of the gene were found in two patients, identifying LIS-1 as the disease gene. The deduced amino-acid sequence shows significant homology to beta-subunits of heterotrimeric G proteins, suggesting that it could possibly be involved in a signal transduction pathway crucial for cerebral development.


Techniques for Non-Invasive Monitoring of Arterial Blood Pressure.
Since both, hypotension and hypertension, can potentially impair the function of vital organs such as heart, brain, or kidneys, monitoring of arterial blood pressure (BP) is a mainstay of hemodynamic monitoring in acutely or critically ill patients. Arterial BP can either be obtained invasively via an arterial catheter or non-invasively. Non-invasive BP measurement provides either intermittent or continuous readings. Most commonly, an occluding upper arm cuff is used for intermittent non-invasive monitoring. BP values are then obtained either manually (by auscultation of Korotkoff sounds or palpation) or automatically (e.g., by oscillometry). For continuous non-invasive BP monitoring, the volume clamp method or arterial applanation tonometry can be used. Both techniques enable the arterial waveform and BP values to be obtained continuously. This article describes the different techniques for non-invasive BP measurement, their advantages and limitations, and their clinical applicability.


Risk factor assessment for the acquisition of fluoroquinolone-resistant isolates of Pseudomonas aeruginosa in a community-based hospital.
A case-control study was performed in a community-based nonteaching hospital to assess patient risk factors for the acquisition of fluoroquinolone-resistant isolates of Pseudomonas aeruginosa. Fifty-five patients who were hospitalized between July 1, 1993 and December 31, 1993 and who had P. aeruginosa recovered from a clinical specimen were included in the analysis. Two patient populations were designated based on the fluoroquinolone susceptibility of their P. aeruginosa isolates. Statistical evaluation using univariate analysis of demographic and clinical data from the 42 patients with quinolone-susceptible P. aeruginosa and the 13 patients with quinolone-resistant P. aeruginosa demonstrated that prior receipt of a fluoroquinolone was the only significant risk factor for the subsequent emergence of fluoroquinolone resistance among P. aeruginosa isolated from patients hospitalized in this small community-based institution (p = 0.0196). Multivariate analysis supported the finding that prior receipt of a fluoroquinolone was the major risk factor for the isolation of fluoroquinolone-resistant P. aeruginosa (p = 0.0004); isolation of this Gram-negative bacillus from sputum (p = 0.0306) and a history of recent surgery (p = 0.0058) were also significantly associated as risk factors for resistance.


First report of neurotoxic effect of the cyanobacterium Cylindrospermopsis raciborskii on the motility of trematode metacercariae.
Cylindrospermopsis raciborskii (Woloszynska) is a photosynthetic cyanobacterium that can produce cytotoxic (cylindrospermopsin) and neurotoxic cyanotoxins (saxitoxins). In Brazil the strains of C. raciborskii are reported to produce only saxitoxins (STX) and their effect on fish parasites has not been tested to date. The fish Poecilia vivipara Bloch and Schneider is a common host for the trematode Pygidiopsis macrostomum Travassos off the coast of Rio de Janeiro, and this fish-parasite interaction is a model for behavioural and ecotoxicological studies. The aim of this work was to evaluate the motility of metacercariae of P. macrostomum from P. vivipara exposed to 40 mg l-1 and 400 mg l-1 of crude lyophilized extract of the cyanobacterium C. raciborskii (CYRF-01) for 48 h. The fish were separated into groups of ten individuals and, after exposure, five fish from each group were dissected for counting and checking the motility of metacercariae. The other five fish were dissected after 48 h in clean water. The detection and quantification of STX in the solutions of cyanobacteria, and the gills and guts of fish, were performed by an enzyme-linked immunosorbent assay. The crude extract of C. raciborskii caused temporary paralysis in metacercariae of P. macrostomum after exposure of fish to both concentrations, and the motility recovered after the fish were kept for 48 h in clean water. STX was detected in the guts and gills of all fish analysed, suggesting that this toxin is involved in the paralysis of metacercariae. This is the first report on the action of neurotoxins in metacercariae of fish.


Obesity-induced hypoadiponectinaemia: the opposite influences of central and peripheral fat compartments.
The substantial reduction in adiponectin concentration among obese individuals seems to depend on fat distribution and is a marker of metabolic and adipose tissue dysfunction. We aimed to: (i) address whether abdominal fat from different compartments (visceral, deep subcutaneous abdominal and superficial subcutaneous abdominal) and gluteofemoral fat are independently associated with blood adiponectin concentration; and (ii) investigate whether abdominal (proxied by waist circumference) and gluteofemoral fat (proxied by hip circumference) accumulation causally determine blood adiponectin concentration. To investigate the independent association of abdominal and gluteofemoral fat with adiponectin concentration, we used multivariable regression and data from 30-year-old adults from the 1982 Pelotas Birth Cohort (n = 2,743). To assess the causal role of abdominal and gluteofemoral fat accumulation on adiponectin concentration, we used Mendelian randomization and data from two consortia of genome-wide association studies-the GIANT (n > 210 000) and ADIPOGen consortia (n = 29 347). In the multivariable regression analysis, all abdominal fat depots were negatively associated with adiponectin concentration, specially visceral abdominal fat [men: β = -0.24 standard unit of log adiponectin per standard unit increase in abdominal fat; 95% confidence interval (CI) = -0.31, -0.18; P = 8*10-13; women: β = -0.31; 95% CI = -0.36, -0.25; P = 7*10-27), whereas gluteofemoral fat was positively associated with adiponectin concentration (men: β = 0.13 standard unit of log adiponectin per standard unit increase in gluteofemoral fat; 95% CI = 0.03, 0.22; P = 0.008; women: β = 0.24; 95% CI = 0.17, 0.31; P = 7*10-11). In the Mendelian randomization analysis, genetically-predicted waist circumference was inversely related to blood adiponectin concentration (β = -0.27 standard unit of log adiponectin per standard unit increase in waist circumference; 95% CI = -0.36, -0.19; P = 2*10-11), whereas genetically-predicted hip circumference was positively associated with blood adiponectin concentration (β = 0.17 standard unit of log adiponectin per standard unit increase in hip circumference; 95% CI = 0.11, 0.24; P = 1*10-7). These results support the hypotheses that there is a complex interplay between body fat distribution and circulating adiponectin concentration, and that whereas obesity-induced hypoadiponectinaemia seems to be primarily attributed to abdominal fat accumulation, gluteofemoral fat accumulation is likely to exert a protective effect.


Biomarkers for serum diagnosis of infectious diseases and their potential application in novel sensor platforms.
Nanotechnological tools and biomarkers for diagnosis and prognosis, as well as strategies for disease control and monitoring populations at higher risk, are continuous worldwide challenges for infectious diseases. Phage display and monoclonal antibody combinatorial libraries are important sources for biomarker discovery and for improved diagnostic strategies. Mimetic peptides were selected against polyclonal antibodies from patients with dengue fever, leprosy, and leishmaniasis as model diseases, and from immunized chickens with total antigens from all three pathogens. Selected single or combined multi-epitope peptide biomarkers were further associated with four different sensor platforms, classified as affinity biosensors, that may be suitable as general protocols for field diagnosis. We have also developed two methods for nanoparticle agglutination assays (a particle gel agglutination test and a magnetic microparticle [MMP]-enzyme-linked immunosorbent assay [ELISA]) and two electrochemical biosensors (impedimetric and amperometric) for DNA and antibody detection. For the agglutination tests, micro- and nanoparticles were coupled with filamentous bacteriophages displaying the selected mimotopes on their surfaces, which has favored the formation of the antigen-antibody or peptide-protein complexes, amplifying the optical detection in ELISA assays or after the chromatographic separation of the microagglutinates. We have also demonstrated a proof-of-concept for the electrochemical biosensors by using electrodes modified with novel functionalized polymers. These electrochemical biosensors have proven to be fast, very sensitive, and specific for the detection of pathogen DNA and circulating antibodies of patients, which may become important in a wide range of diagnostic devices for many infectious agents.


To Do or Not to Do? Task Control Deficit in Obsessive-Compulsive Disorder.
Task control is an executive control mechanism that facilitates goal-directed task selection by suppressing irrelevant automatic "stimulus-driven" behaviors. In the current study, we test the hypothesis that less efficient task control in individuals diagnosed with obsessive-compulsive disorder (OCD) is associated with OCD symptoms, and specifically, with the inability to inhibit unwanted behaviors in OCD. Thirty-five healthy controls, 30 participants with OCD, and 26 participants with generalized anxiety disorder (GAD) completed the object-interference (OI) task to measure task control, the stop-signal task to measure response inhibition, and the arrow-flanker task to evaluate executive abilities not contingent upon task control. OCD patients, but not GAD patients or healthy controls, exhibited impaired performance on the OI task. The deficit in task control, but not in response inhibition, correlated with OCD symptom severity. We suggest that reduced task control may be one of the neurocognitive processes that underlie the inability to inhibit unwanted behaviors in OCD.


Fibropapillomatosis and the Chelonid Alphaherpesvirus 5 in Green Turtles from West Africa.
Fibropapillomatosis (FP) is a tumorigenic panzootic disease of sea turtles, most common in green turtles (Chelonia mydas). FP is linked to the chelonid alphaherpesvirus 5 (ChAHV5) and to degraded habitats and, though benign, large tumours can hinder vital functions, causing death. We analyse 108 green turtles, captured in 2018 and 2019, at key foraging grounds in Guinea-Bissau and Mauritania, West Africa, for the presence of FP, and use real-time PCR to detect ChAHV5 DNA, in 76 individuals. The prevalence of FP was moderate; 33% in Guinea-Bissau (n = 36) and 28% in Mauritania (n = 72), and most turtles were mildly affected, possibly due to low human impact at study locations. Juveniles had higher FP prevalence (35%, n = 82) compared to subadults (5%, n = 21), probably because individuals acquire resistance over time. ChAHV5 DNA was detected in 83% (n = 24) of the tumour biopsies, consistent with its role as aetiological agent of FP and in 26% (n = 27) of the 'normal' skin (not showing lesions) from FP turtles. Notably, 45% of the asymptomatic turtles were positive for ChAHV5, supporting multifactorial disease expression. We report the first baselines of FP and ChAHV5 prevalence for West Africa green turtles, essential to assess evolution of disease and future impacts of anthropogenic activities.


Determination of band edges and their influences on photocatalytic reduction of nitrobenzene by bulk and exfoliated g-C3N4.
Thermally and chemically exfoliated metal-free semiconducting g-C3N4 are synthesized from bulk g-C3N4. Thorough characterization of the synthesized materials is performed with the help of XRD, FTIR, FE-SEM, PL, surface area analysis and DRS to probe differences in structural, morphological and optical properties between thermally and chemically exfoliated g-C3N4. The synthesized materials are exposed to light for photocatalytic reduction of nitrobenzene. The complete reduction reaction mechanism and product selectivity over the synthesized catalysts are studied in this report. The rate of reduction of nitrobenzene is found to be higher with thermally exfoliated g-C3N4, and the selectivity of aniline is found to be higher in the case of chemical exfoliated g-C3N4. The differences in the reactivity are explained in terms of structure, surface morphologies and band edge positions.


Pd/Xu-Phos-catalyzed asymmetric elimination of fully substituted enol triflates into axially chiral trisubstituted allenes.
The β-H elimination, as one of the most important elementary reactions in transition metal chemistry, is a key step in quenching the carbon-palladium bond for the Heck reaction. However, the β-H elimination of the alkenyl palladium species leading to allene is an energetically unfavored process, and therefore, it has been a long-standing challenge in control of this process via enantioselective manner. We developed a concise and efficient methodology to construct trisubstituted chiral allenes from stereodefined fully substituted enol triflates by the enantioselective β-H elimination of the alkenyl palladium species under mild conditions. The identified Xu-Phos play a crucial role in the chemoselectivity and enantioselectivity. Multiple linear regression analysis shows the important steric effect on enantioselectivity. DFT computation results allow us to propose an intramolecular base (-OAc)-assisted deprotonation mechanism for this progress. Distortion-interaction and energy decomposition analysis indicate that the difference in electrostatic energy (Eelec) of the two intramolecular base-assisted deprotonation transition states dominates the stereoselectivity.


Orthopaedics and the law.
Understanding the relevant legal context is critical to the safe and successful practice of orthopaedic surgery. Specifically, three areas of liability are relevant to most physicians: medical malpractice, products liability, and the liability of health care organizations. Medical malpractice encompasses the professional physician-patient relationship with its implied contract, consent, fiduciary responsibilities, and duty to provide the standard of care, as well as certain common-law duties pertinent in special circumstances. Orthopaedic surgeons who design implants or who have a relationship with a device manufacturer are at risk for liability for a failed product. In general, the hospital entity is responsible for the actions of its physician-employees. Still unclear is the degree to which a physician is obligated to appeal to a third-party payer on behalf of a patient. Physicians should remember that, above all else, common sense with regard to the treatment, informed consent, and advocacy of patients is essential to avoiding many medical-legal pitfalls.


Prevalence of Parental Smoking and Predictors of Cessation: A Study in the South Carolina Pediatric Practice Research Network.
Secondhand smoke exposure harms children. The objectives of the study were to determine the prevalence of secondhand smoke exposure in children ≤2 years and determine the predictors of smoking and smoking cessation in parents. We surveyed parents of children ≤2 years of age, asking about parental smoking patterns, interest in quitting and children's respiratory symptoms. Data were analyzed with chi-square and multiple logistic regression. Thirteen percent were current smokers and 18% had quit. The most common reason for quitting was being pregnant (42%). Children's respiratory symptoms did not predict quitting. Parents on Medicaid were more likely to smoke than those on private insurance (OR = 5.7, 95% CI = 2.0-16.5) and less likely to quit (OR = 0.2, 95% CI = 0.1-0.9). Having a new baby may be a motivator for parents to quit. We must address socioeconomic factors to develop a successful intervention in pediatric practices.


Prostaglandin E2 gel compared to oxytocin for medically-indicated labour induction at term: a controlled clinical trial.
A clinical trial was carried out to compare the efficacy and tolerance of two modes of induction of labour: vaginally administered prostaglandin E2 gel vs intravenous perfusion of synthetic oxytocin. Fifty women with pregnancy at or near term in whom prompt vaginal delivery was clinically indicated were divided at random into sub-groups of 25 each. Initial dose of PGE2 gel was 1 mg followed by another application of 1 mg or 2 mg 6 hours later in case active labour stage has not been reached. Progressive oxytocin perfusion began with 1 mU/min., being increased gradually every 20 minutes until efficacious uterine dynamics were attained. Data were recorded on entry of age, parity, gestation age, cervical dilatation, Bishop score, indication for induction. Continuous materno-foetal monitoring was carried out during the induction period. Results were evaluated from induction/delivery time, type of delivery, maternal and foetal condition, and any side-effects which developed. Apart from a higher number of instrumental deliveries in the PGE2 gel series, which was not related to the induction method, there was no significant difference between any of the variables evaluated, both methods producing active labour in approximately 70% of the patients within 12 hours. The authors stress, however, the convenience, both for patients and hospital staff, of the administration of an intravaginal induction agent over a systemic therapeutic method of induction.


Heparin-bridging therapy and risk of post-polypectomy bleeding: Meta-analysis of data reported by Japanese colonoscopists.
Peri-procedural bridging (PPB) with heparin is recommended for patients with high thromboembolic risk who need to withhold antithrombotic therapy for colonoscopic polypectomy. However, little is known about the bleeding risk from heparin-bridging therapy itself. MEDLINE and EMBASE databases were searched through January 2017 for studies that compared the risk of PPB in patients who received heparin-bridging therapy in lieu of antithrombotic agents for colonoscopic polypectomy and those who discontinued antithrombotic agents without receiving heparin. Pooled odds ratio (OR) and 95% confidence interval (CI) were calculated using a random-effects model, generic inverse variance method. Between-study heterogeneity was quantified using the Q statistic and I2 . A total of five studies consisting of 2601 patients were identified. A significantly increased risk of PPB among bridged patients compared to non-bridged patients was demonstrated with a pooled OR of 8.29 (95% CI, 4.96-13.87). Statistical heterogeneity was low with I2 of 0%. The present study demonstrated a significantly increased risk of PPB among patients who underwent colonoscopic polypectomy and received heparin-bridging therapy in lieu of antithrombotic agents compared to patients who did not receive it.


Quirks of Error Estimation in Cross-Linking/Mass Spectrometry.
Cross-linking/mass spectrometry is an increasingly popular approach to obtain structural information on proteins and their complexes in solution. However, methods for error assessment are under current development. We note that false-discovery rates can be estimated at different points during data analysis, and are most relevant for residue or protein pairs. Missing this point led in our example analysis to an actual 8.4% error when 5% error was targeted. In addition, prefiltering of peptide-spectrum matches and of identified peptide pairs substantially improved results. In our example, this prefiltering increased the number of residue pairs (5% FDR) by 33% (n = 108 to n = 144). This number improvement did not come at the expense of reduced accuracy as the added data agreed with an available crystal structure. We provide an open-source tool, xiFDR ( https://github.com/rappsilberlab/xiFDR ), that implements our observations for routine application. Data are available via ProteomeXchange with identifier PXD004749.


Tile-Based Pairwise Analysis of GC × GC-TOFMS Data to Facilitate Analyte Discovery and Mass Spectrum Purification.
A new tile-based pairwise analysis workflow, termed 1v1 analysis, is presented to discover and identify analytes that differentiate two chromatograms collected using comprehensive two-dimensional (2D) gas chromatography coupled with time-of-flight mass spectrometry (GC × GC-TOFMS). Tile-based 1v1 analysis easily discovered all 18 non-native analytes spiked in diesel fuel within the top 30 hits, outperforming standard pairwise chromatographic analyses. However, eight spiked analytes could not be identified with multivariate curve resolution-alternating least-squares (MCR-ALS) nor parallel factor analysis (PARAFAC) due to background contamination. Analyte identification was achieved with class comparison enabled-mass spectrum purification (CCE-MSP), which obtains a pure analyte spectrum by normalizing the spectra to an interferent mass channel (m/z) identified from 1v1 analysis and subtracting the two spectra. This report also details the development of CCE-MSP assisted MCR-ALS, which removes the identified interferent m/z from the data prior to decomposition. In total, 17 out of 18 spiked analytes had a match value (MV) > 800 with both versions of CCE-MSP. For example, MCR-ALS and PARAFAC were unable to decompose the pure spectrum of methyl decanoate (MVs < 200) due to its low 2D chromatographic resolution (∼0.34) and high interferent-to-analyte signal ratio (∼30:1). By leveraging information gained from 1v1 analysis, CCE-MSP and CCE-MSP assisted MCR-ALS obtained a pure spectrum with an average MV of 908 and 964, respectively. Furthermore, tile-based 1v1 analysis was applied to track moisture damage in cacao beans, where 86 analytes with at least a 2-fold concentration change were discovered between the unmolded and molded samples. This 1v1 analysis workflow is beneficial for studies where multiple replicates are either unavailable or undesirable to save analysis time.


[Profile of adults suffering from COPD in Poland].
Chronic obstructive pulmonary disease (COPD) is one of the most common chronic diseases in adults. It is estimated, that in Poland around two million people suffer from COPD. The aim of this study, was to characterize population of patients with COPD in Poland. The study, established and coordinated by the Polish Respiratory Society, included a representative sample of 500 GPs where were asked to fill questionnaires on diagnosis and treatment of their COPD patients. The questions dealt with disease history and clinical presentation, COPD severity stage, diagnostic and therapeutic procedures. Altogether 298 physicians (59.6% of invited) provided information about 2756 COPD patients aged 61.6 ± 11.1 years (36.3% were women). According to GOLD recommendations 16.6% of patients had mild, 57.0% moderate, 18.6% severe and 2.1% very severe COPD. Smoking history was declared by 97.8% of respondents. 51.4% of COPD patients had continued smoking. Over the last year Ambulance Service intervened in 19.7% of patients and 29.1% of respondents required hospital treatment of COPD. Among more than 80% of patients, doctor diagnosed limitation in exercise tolerance, and shortness of breath at rest, and in approximately 60% of the respondents were presented productive cough, weakened vesicular murmur and prolonged phase of exhalation. Despite the diagnosis, more than half of men and women had continued smoking. The number of hospitalizations and emergency intervention positively correlated with the severity of the disease. The survey results emphasize the urgent need for health education in patients with COPD.


Corticosteroid therapy in asthma.
Corticosteroids are useful in the treatment of both allergic and idiosyncratic asthma. Although the mechanisms of corticosteroid action in asthma are poorly understood, several possible sites of action have been proposed. Corticosteroids alter the cellular and vascular inflammatory response to bronchial injury, affect catecholamine action on airways, and alter the production of eicosanoids, all of which aid in the resolution of bronchospasm in asthmatic patients. Corticosteroids should only be used for the treatment of asthma after therapeutic levels of methylxanthines and beta agonists have been achieved. Although the optimal doses of corticosteroids in asthma have not been defined, guidelines exist to aid in therapy. In the treatment of status asthmaticus, the intravenous route of administration is preferable. Short courses of corticosteroids may be useful in the treatment of chronic asthma. When long-term corticosteroid therapy is the only option for control of bronchospasm, alternate-day and/or aerosolized corticosteroids are preferable to daily corticosteroids and are associated with fewer side effects. Corticosteroids are useful in the pregnant asthmatic patient when bronchospasm cannot be controlled with bronchodilators. The major risk to the fetus in pregnant asthmatics is hypoxia from uncontrolled bronchospasm, and not from therapy. However, the lowest possible dose of systemic corticosteroids needed to control symptoms, with or without the use of aerosolized corticosteroids, is recommended. All asthmatics who have needed systemic or aerosolized corticosteroids within 6 months prior to surgery should receive preoperative and post-operative corticosteroid therapy. For patients not usually on systemic corticosteroids, conversion to oral prednisone, with a rapid taper is recommended. Side effects from short-term corticosteroid therapy are minimal, with hyperglycemia and psychosis being the major concerns. Long-term steroid therapy has significant side effects, however, and use should be minimized. Suppression of the HPA axis is one of the most potentially dangerous side effects of corticosteroids, and therefore any patient who has been treated with corticosteroids for longer than 4 weeks should be evaluated for possible adrenal suppression.


Pyrimethamine (2,4-diamino-5-p-chlorophenyl-6-ethylpyrimidine) induces apoptosis of freshly isolated human T lymphocytes, bypassing CD95/Fas molecule but involving its intrinsic pathway.
Pyrimethamine (2,4-diamino-5-p-chlorophenyl-6-ethyl-pyrimidine), a folic acid antagonist, may exert, in addition to antiprotozoan effects, immunomodulating activities, including induction of peripheral blood lymphocyte apoptosis. However, the molecular mechanisms underlying this proapoptotic activity remain to be elucidated. Here we show that pyrimethamine, used at a pharmacologically relevant concentration, induced per se apoptosis of activated lymphocytes via the activation of the caspase-8- and caspase-10-dependent cascade and subsequent mitochondrial depolarization. Importantly, this seems to occur independently from CD95/Fas engagement. The proapoptotic activity of pyrimethamine was further confirmed in a patient with autoimmune lymphoproliferative syndrome, an immune disorder associated with a defect of Fas-induced apoptosis. In this patient, pyrimethamine treatment resulted in a "normalization" of lymphocyte apoptosis with a significant amelioration of laboratory parameters. Altogether, these results suggest a mechanism for pyrimethamine-mediated apoptosis that seems to bypass CD95/Fas engagement but fully overlaps CD95/Fas-induced subcellular pathway. On these bases, a reappraisal of the use of pyrimethamine in immune lymphoproliferative disorders characterized by defects in CD95/Fas-mediated apoptosis should be taken into account.


Repair of osteochondral defects: efficacy of a tissue-engineered hybrid implant containing both human MSC and human iPSC-cartilaginous particles.
Both mesenchymal stromal cells (MSC) and induced pluripotent stem cells (iPSC) offer the potential for repair of damaged connective tissues. The use of hybrid implants containing both human MSC and iPSC was investigated to assess their combined potential to yield enhanced repair of osteochondral defects. Human iPSC-CP wrapped with tissue engineered constructs (TEC) containing human MSC attained secure defect filling with good integration to adjacent tissue in a rat osteochondral injury model. The presence of living MSC in the hybrid implants was required for effective biphasic osteochondral repair. Thus, the TEC component of such hybrid implants serves several critical functions including, adhesion to the defect site via the matrix and facilitation of the repair via live MSC, as well as enhanced angiogenesis and neovascularization. Based on these encouraging studies, such hybrid implants may offer an effective future intervention for repair of complex osteochondral defects.


Germanium Dicarbide: Evidence for a T-Shaped Ground State Structure.
The equilibrium structure of germanium dicarbide GeC2 has been an open question since the late 1950s. Although most high-level quantum calculations predict an L-shaped geometry, a T-shaped or even a linear geometry cannot be ruled out because of the very flat potential energy surface. By recording the rotational spectrum of this dicarbide using sensitive microwave and millimeter techniques, we unambiguously establish that GeC2 adopts a vibrationally averaged T-shaped structure in its ground state. From analysis of 14 isotopologues, a precise r0 structure has been derived, yielding a Ge-C bond length of 1.952(1) Å and an apex angle of 38.7(2)°.


[Synergistic cytotoxic effects of chemotherapy in colon tumor cells by simultaneous inhibition of de novo and salvage energy metabolism pathways].
The success of chemotherapy of colon tumours is currently limited. We have therefore used the human colon tumour cell line HT-29 to evaluate the cytotoxic effects of various drug combinations. Trimidox (3,4,5-trihydroxybenzamidoxime), a recently patented inhibitor of ribonucleotide reductase was combined with cytosinearabinoside (Ara-C) or 2',2'-difluorodeoxycytidine (DFDC) in order to inhibit both pyrimidine de novo and salvage pathways. Synergistic cytotoxic effects were observed. When HT-29 cells were sequentially treated with trimidox (20 microM for 24 h) and Ara-C (2 microM for 2 h), colony numbers decreased to 71% of the value calculated for additive cytotoxicity. When cells were simultaneously treated with trimidox (10 microM and 15 microM) and DFDC (0.2 nM), synergistic inhibition of colony formation was likewise noted (colony numbers decreased to values as low as 73% or 71% of the values calculated for additive cytotoxicity). On the other hand, we combined tiazofurin, an inhibitor of the guanylate de novo pathway, with allopurinol, which inhibits the guanylate salvage pathway by increasing intracellular hypoxanthine concentrations, leading to inhibition of the enzyme hypoxanthine-guanine phosphoribosyltransferase (HGPRT). Synergistic cytotoxic effects were observed under these conditions too. When cells were treated with 10 microM tiazofurin and 400 microM or 800 microM allopurinol the number of colonies decreased to 69% and 27%, respectively, of the values calculated for additive effects. Our data suggest these drug combinations to be promising options in the treatment of human colon cancer.


Traumatic Experience and Somatoform Dissociation Among Spirit Possession Practitioners in the Dominican Republic.
Recent studies in African contexts have revealed a strong association between spirit possession and severe trauma, with inclusion into a possession cult serving at times a therapeutic function. Research on spirit possession in the Dominican Republic has so far not included quantitative studies of trauma and dissociation. This study evaluated demographic variables, somatoform dissociative symptoms, and potentially traumatizing events in the Dominican Republic with a group of Vodou practitioners that either do or do not experience spirit possession. Inter-group comparisons revealed that in contrast to non-possessed participants (n = 38), those experiencing spirit possession (n = 47) reported greater somatoform dissociation, more problems with sleep, and previous exposure to mortal danger such as assaults, accidents, or diseases. The two groups did not differ significantly in other types of trauma. The best predictor variable for group classification was somatoform dissociation, although those items could also reflect the experience of followers during a possession episode. A factor analysis across variables resulted in three factors: having to take responsibility early on in life and taking on a professional spiritual role; traumatic events and pain; and distress/dissociation. In comparison with the non-possessed individuals, the possessed ones did not seem to overall have a remarkably more severe story of trauma and seemed to derive economic gains from possession practice.


Anomalously low friction of confined monolayer water with a quadrilateral structure.
In this work, we explored how the structure of monolayer water confined between two graphene sheets is coupled to its dynamic behavior. Our molecular dynamics simulations show that there is a remarkable interrelation between the friction of confined water with two walls and its structure under extreme confinement. When the water molecules formed a regular quadrilateral structure, the friction coefficient is dramatically reduced. Such a low-friction coefficient can be attributed to the formation of long-range ordered hydrogen bond network, which not only decreases the structure corrugation in the direction perpendicular to the walls but also promotes the collective motion of the confined water. The regular quadrilateral structure can be formed only if the number density of confined water falls within a certain range. Higher number density results in larger structure corrugations, which increases the friction, while smaller number density leads to an irregular hydrogen bond network in which the collective motion cannot play the role. We demonstrated that there are four distinct stages in the diagram of the friction coefficient vs the number density of confined water. This research clearly established the connection between the dynamic characteristics of confined monolayer water and its structure, which is beneficial to further understand the mechanism of the high-speed water flow through graphene nanocapillaries observed in recent experiments.


Reduced delivery of epididymal adipocyte-derived exosomal resistin is essential for melatonin ameliorating hepatic steatosis in mice.
Adipocyte-derived exosomes (Exos) serve as bioinformation-containing messengers in cell-to-cell communications, and numerous reports demonstrate that resistin, an adipokine, is strongly associated with hepatic steatosis and other fatty liver diseases, suggesting that adipose dysfunction-generated altered pattern of exosomal cytokines may contribute to shaping the physiological activities in liver. Admittedly, melatonin-mediated positive effects on various tissues/organs have been respectively reported, but regulatory mechanisms of melatonin on the crosstalk between adipose tissue and liver have been investigated rarely. Overall, we hypothesize that the crosstalk originating from adipose tissue may be another worthy regulatory pathway for melatonin ameliorating of hepatic steatosis. Here, we first found the amount of adipocyte-derived exosomal resistin to be significantly decreased by melatonin supplementation. Compared to mice with ExosHFD or Exosresistin treatment, ExosMT remarkably ameliorated hepatic steatosis. Further test demonstrated that resistin was a pivotal cytokine which repressed phosphorylation of 5' adenosine monophosphate-activated protein kinase α (pAMPKα Thr172 ) to trigger endoplasmic reticulum (ER) stress, resulting in hepatic steatosis, whereas ExosMT reversed these risks in hepatocytes. In adipocytes, we identified melatonin to reduce the production of resistin through the brain and muscle arnt-like protein 1 (Bmal1) transcriptional inhibition. Notably, we also confirmed that melatonin enhanced N6 -Methyladenosine (m6 A) RNA demethylation to degrade resistin mRNA in adipocytes. Overall, melatonin decreases traffic volume of adipocyte-generated exosomal resistin from adipocytes to hepatocytes, which further alleviates ER stress-induced hepatic steatosis. Our findings illustrate a novel melatonin-mediated regulatory pathway from adipocytes to hepatocytes, indicating that adipocyte-derived exosome is a new potential target for treating obesity and related hepatorenal syndrome.


Double-blind study of a total liver extract in patients with hepatic dysfunction.
Forty hospitalized male patients with hepatic functional deficit were treated i.m. for 3 weeks with a total liver extract or placebo. The study was a double-blind. Statistical analysis (Student's t test, analysis of variance. Mann-Whitney test and Fisher's probability test) showed that liver extract therapy was more effective than placebo in improving liver function as indicated by changes in clinical features(digestive disorders, constipation, hepatomegaly) and colloidal and other laboratory tests, e.g. plasma cholesterol, prothrombin time, red cell count and hemoglobin concentration. An overall clinical assessment showed that 60% of patients treated with the liver extract were improved. In addition, no local or general side effects were observed. Present results are in favour of an hepatoprotective activity of a total liver extract.


The isolation, identification and molecular analysis of Candida spp. isolated from the oral cavities of patients with diabetes mellitus.
Previous studies have shown a high incidence (77%) of isolation of Candida spp. from the oral cavities of patients with type 1 diabetes mellitus. The aim of the present study was to assess the prevalence of yeast in the oral cavities of patients suffering from type 1 and type 2 diabetes mellitus. The patients were classified according to the level of diabetic control (HbA1c), and further stratified on the presence or absence of dental prosthesis. Oral rinse samples were assessed for the growth of yeast and the degree of colonization. Oral isolates were defined to the species level by both phenotypic and novel molecular methods. The overall proportion (60%) of diabetic patients who had Candida spp. isolated from the oral cavity was similar to that previously reported. Local oral factors, such as the presence of dentures, seemed to have a greater influence than diabetic status on the amount and species of Candida isolated from the oral cavities of diabetic patients. Diabetic patients with dentures had more non-albicans Candida isolated from their mouths than dentate diabetic patients. Candida dubliniensis was isolated from diabetic patients and may have a predilection for dentate patients.


18F-fluorodeoxyglucose (FDG) PET or 18F-fluorothymidine (FLT) PET to assess early response to aromatase inhibitors (AI) in women with ER+ operable breast cancer in a window-of-opportunity study.
This study evaluated the ability of 18F-Fluorodeoxyglucose (FDG) and 18F-Fluorothymidine (FLT) imaging with positron emission tomography (PET) to measure early response to endocrine therapy from baseline to just prior to surgical resection in estrogen receptor positive (ER+) breast tumors. In two separate studies, women with early stage ER+ breast cancer underwent either paired FDG-PET (n = 22) or FLT-PET (n = 27) scans prior to endocrine therapy and again in the pre-operative setting. Tissue samples for Ki-67 were taken for all patients both prior to treatment and at the time of surgery. FDG maximum standardized uptake value (SUVmax) declined in 19 of 22 lesions (mean 17% (range -45 to 28%)). FLT SUVmax declined in 24 of 27 lesions (mean 26% (range -77 to 7%)). The Ki-67 index declined in both studies, from pre-therapy (mean 23% (range 1 to 73%)) to surgery [mean 8% (range < 1 to 41%)]. Pre- and post-therapy PET measures showed strong rank-order agreement with Ki-67 percentages for both tracers; however, the percent change in FDG or FLT SUVmax did not demonstrate a strong correlation with Ki-67 index change or Ki-67 at time of surgery. A window-of-opportunity approach using PET imaging to assess early response of breast cancer therapy is feasible. FDG and FLT-PET imaging following a short course of neoadjuvant endocrine therapy demonstrated measurable changes in SUVmax in early stage ER+ positive breast cancers. The percentage change in FDG and FLT-PET uptake did not correlate with changes in Ki-67; post-therapy SUVmax for both tracers was significantly associated with post-therapy Ki-67, an established predictor of endocrine therapy response.


Genomic approach to biomarker identification and its recent applications.
This paper discusses selected activities, issues, and challenges in recent development of analytical methods and applications in biomarker identification and validation using state-of-the-art genomic approaches. Molecular profiling via genomics, proteomics, and metabonomics has opened new windows to study disease states and biological systems. It has also provided exciting opportunities for novel applications in clinical research as well as in drug discovery and development. In the past several years, we have witnessed enormous progress resulting particularly from gene expression profiling of mRNA or transcriptomics. After a brief review on technology advances in gene expression profiling using microarrays, I mainly discuss recent developments of the genomic approaches to biomarker identification and validation in two major types of applications. The first type involves examples in cancer diagnostics and prognostics based on tumor gene expression profiling, whereas the second type involves biomarker applications in drug discovery and development. The focus will be on analytical methods and algorithms that have been developed in recent years facilitating biomarker discovery and application by leveraging genome-wide expression profiles derived from microarrays. Technical issues in experimental design, data processing, error modeling, quality control, figures of merit for performance evaluation, and meta-analysis related to biomarker discovery and application are also discussed. A case study of disease outcome prognosis for breast cancer patients based on tumor expression pattern is presented before closing remarks.


Congenital neural hamartoma ("fascicular schwannoma"). A light microscopic, immunohistochemical, and ultrastructural study.
A previously undescribed form of a congenital neural hamartoma composed entirely of Schwann cells in a fascicular pattern was found on the leg of a male infant. The lesion was thought to be an unusual variant of plexiform Schwannoma or a newly recognized unencapsulated form of Schwannoma. On light microscopic examination, the lesion, which measured 5 x 4 cm when it was surgically removed when the infant was 7 months old, showed an unencapsulated dermal mass composed of fascicles of spindle cells with frequent Verocay body-like structures. The intervening stroma was collagenous and contained an increased number of mast cells. Special stains did not demonstrate any axons in the tumor. There was a strongly positive immunohistochemical reaction for S-100 protein and collagen type IV in the spindle cells. These cells were weakly or focally positive for Leu-7 and vimentin, and completely negative for neural filaments, neuron-specific enolase, glial fibrillary acidic protein, epithelial membrane antigen, desmin, and muscle-specific actin. On electron microscopic examination, the spindle cells were found to be surrounded by basal lamina and showed frequent cytoplasmic invagination filled with collagen bundles. No unmyelinated nerve fibers were identified.


Biosurfactant production by yeasts from different types of soil of the South Shetland Islands (Maritime Antarctica).
To screen and identify a potential biosurfactant-producing yeast strain isolated from Antarctic soil and to evaluate the fermentation process kinetics of the most promising strain on biosurfactant production using glycerol as carbon source. From the 68 isolated yeast strains, 11 strains were able to produce biosurfactants after Emulsification Index (E.I.) and Drop Collapse tests, reaching an E.I. higher than 10%. Strain 1_4.0 was the best producer, identified as Candida glaebosa based on molecular analysis. Yeast was cultivated in a medium composed of glycerol supplemented with yeast extract for 120 h to determine the process kinetics. The increased C/N ratio affected yeast growth and biosurfactant production. Biosurfactant release was associated with the end of exponential and beginning of the stationary growth phases. Results indicated an E.I. of 30% at the end of the fermentation. The feasiability of C. glaebosa to produce biosurfactant from a low-cost medium cultivation shows a great impact on the development of bioresource in the Antarctica terrestrial environment. Although the diversity of psychrophilic/psychrotolerant micro-organisms from Antarctica has been the preferred subject of study by microbiologists, terrestrial microfungal communities are scarcely investigated and literature about the biotechnological potential of such micro-organisms should cover important biomolecules in addition to cold-adapted enzymes. In the present study, for the first time, the Maritime Antarctica environment was screened as a novel source of biosurfactants produced by micro-organisms.


Vitamin D Receptor Gene Polymorphisms and Cigarette Smoking Impact on Oral Health: A Case-Control Study.
Periodontal diseases are multiperspective problems resulting from numerous and diverse exposures that influence the process of initiation or progression of disease. The negative influence of tobacco smoking on oral health is well documented. The aim of the study was to analyze three SNPs in vitamin D receptor gene-rs7975232 (ApaI), rs2228570 (FokI) and rs1544410 (BsmI)-combined with oral health assessment-pH, gingival index, dry mouth, periodontitis, dry socket, redness of oral cavity mucosa, leukoplakia-in a group of cigarette smokers and in non-smokers. Moreover, the possibility of interactions between these polymorphisms and smoking was examined. When comparing the smokers and non-smokers groups, we noticed that rs1544410 heterozygotes were significantly more frequent in the first group, and for the second, both homozygotes were more frequent. Additionally, we observed the impact of interaction between the rs7975232 genotype and smoking status on gingival index. Current smoking was also associated with all analyzed oral health measures except for leucoplakia. Correlation between pH and age in both smokers and non-smokers was also present. Results of our analysis indicate that in our study group lifestyle and aging were leading factors associated with worse oral health status. However, the impact of genetic variants, and also the impact of their interaction with smoking on analyzed parameters was also visible. These results show great possibilities for all levels of prevention of oral diseases by means of education based on evidence-based medicine, but also for incorporating genetic testing and early interventions into this process for predisposed individuals.


Recognizing impossible object relations: intuitions about support in chimpanzees (Pan troglodytes).
Using looking-time measures, the authors examined untrained chimpanzees' (Pan troglodytes) ability to distinguish between adequate and inadequate support. In 3 experiments, the chimpanzees' sensitivity to different support relations between 2 objects was assessed. In each experiment, the chimpanzees saw a possible and an impossible test event, presented as digital video clips. Looking times in the 3 experiments suggest that chimpanzees use amount of contact between 2 objects, but not type of contact, to distinguish between adequate and inadequate support relations. These results indicate that chimpanzees have some intuition about support phenomena but their sensitivity to relational object properties may differ from that of human infants (Homo sapiens) in this domain.


Effects of recombinant imperatoxin A (IpTxa) mutants on the rabbit ryanodine receptor.
Imperatoxin A (IpTxa), a 3.7 kDa peptide from the African scorpion Pandinus imperator, is an agonist of the skeletal muscle ryanodine receptor (RyR1). In order to study the structure of the toxin and its effect on RyR1, IpTxa cDNA was PCR-amplified using 3 pairs of primers, and the toxin was expressed in E. coli. The toxin was further purified by chromatography, and various point mutants in which basic amino acids were substituted by alanine were prepared by site-directed mutagenesis. Studies of single channel properties by the planar lipid bilayer method showed that the recombinant IpTxa was identical to the synthetic IpTxa with respect to high-performance liquid chromatography mobility, amino acid composition and specific effects on RyR1. Mutations of certain basic amino acids (Lys19, Arg23, and Arg33) dramatically reduced the capacity of the peptide to activate RyRs. A subconductance state predominated when Lys8 was substituted with alanine. These results suggest that some basic amino acid residues in IpTxa are important for activation of RyR1, and that Lys8 plays an important role in regulating the gating mode of RyR1.


Nothing really changed: Arizona patient experience of methadone and buprenorphine access during COVID.
To understand patient experience of federal regulatory changes governing methadone and buprenorphine (MOUD) access in Arizona during the COVID-19 pandemic. This community-based participatory and action research study involved one-hour, audio-recorded field interviews conducted with 131 people who used methadone and/or buprenorphine to address opioid use disorder at some point during COVID (January 1, 2020- March 31, 2021) in Arizona. Transcribed data were analyzed using a priori codes focused on federally recommended flexibilities governing MOUD access. Data were quantitated to investigate associations with COVID risk and services access. Telehealth was reported by 71.0% of participants, but the majority were required to come to the clinic to attend video appointments with an offsite provider. Risk for severe COVID outcomes was reported by 40.5% of the sample. Thirty-eight percent of the sample and 39.7% of methadone patients were required to be at the clinic daily to get medication and 47.6% were at high risk for COVID severe outcomes. About half (54.2%) of methadone patients indicated that some form of multi-day take home dosing was offered at their clinic, and 45.8% were offered an extra day or two of multi-day doses; but no participants received the federally allowed 14- or 28-day methadone take-home doses for unstable and stable patients respectively. All participants expressed that daily clinic visits interrupted their work and home lives and desired more take-home dosing and home delivery options. MOUD patients in Arizona were not offered many of the federally allowed flexibilities for access that were designed to reduce their need to be at the clinic. To understand the impact of these recommended treatment changes in Arizona, and other states where they were not well implemented, federal and state regulators must mandate these changes and support MOUD providers to implement them.


Effect of intragastric barostat bag on proximal and distal gastric accommodation in response to liquid meal.
The barostat is the gold standard for measurement of proximal gastric accommodation. Ultrasonography can be used to measure gastric volume. The aim was to investigate the effects of the barostat bag on gastric accommodation and transpyloric flow. Accommodation after a liquid meal (300 ml, 450 kcal) was measured twice at random in eight healthy volunteers. Proximal accommodation was measured once using barostat and once using ultrasound (US). Antrum accommodation was measured using US. Bag volume (BV), antral area (AA), proximal gastric area, and proximal gastric diameter (PGD) data were assessed before and 1, 5, 15, 30, 40, 50, and 60 min postprandially. Transpyloric flow was measured using Doppler 1-5 min postprandially. Fasted, AA size was not affected by the barostat bag (1 mmHg > minimal distension pressure; 2.7 +/- 0.5 vs. 2.6 +/- 0.3 cm(2)). Postprandially, AAs were larger with the bag present (ANOVA, P < 0.04). Maximum AA was reached with the bag in 5 min, without the bag in 1 min postprandially (15.1 +/- 2.3 vs. 9.4 +/- 1.5 cm(2); P < 0.03). Furthermore, AAs were related to BVs (r = 0.57; P < 0.01). After bag deflation, AA decreased (11.9 +/- 1.8 to 7.0 +/- 0.9 cm(2); P = 0.02) and was comparable with the 60-min AA size without the bag (7.1 +/- 1.2 cm(2); P = 0.76) present. Proximal gastric radius calculated from the BVs and PGDs was larger with the bag present (ANOVA, P < 0.001). No effect on early gastric emptying was observed. Postprandially, the barostat bag causes dilatation of the antrum due to meal displacement without influencing early gastric emptying. This antral dilatation is likely to induce exaggerated proximal gastric relaxation observed in studies using the barostat to evaluate fundic accommodation.


Occurrence of neurotensinlike immunoreactivity in subpopulations of hypothalamic, mesencephalic, and medullary catecholamine neurons.
By using indirect immunofluorescence histochemistry combined with the elution-restaining technique, the presence of a neurotensinlike peptide in some catecholamine neurons in the rat brain has been demonstrated. At the level of the medulla oblongata neurotensinlike immunoreactivity was observed in most of the small-sized catecholamine (adrenaline) cell bodies in the dorsolateral part of the nucleus of the solitary tract and in some catecholamine (noradrenaline) cells in the medial part. Neurotensin-positive fibers were found throughout the solitary tract nucleus with increasing concentrations in the rostral direction. Very few neurotensin fibers were seen in the vagal dorsal motor nucleus, which contained a dense network of adrenaline fibers. In the ventral mesencephalon, neurotensinlike immunoreactivity was seen mainly in dopamine cell bodies in the ventral tegmental area, including midline structures, with only single examples of coexistence in the substantia nigra. The dopamine cell bodies of both the A9 and A10 cell groups were surrounded by dense to medium-dense networks of neurotensin fibers. In the hypothalamus numerous dopamine neurons in the arcuate nucleus exhibited neurotensinlike immunoreactivity. Neurotensin-positive nerve terminals, partially overlapping catecholamine (mainly dopamine) fibers, were seen in the external layer of the median eminence. The present results demonstrate coexistence of neurotensinlike immunoreactivity and catecholamines in populations of neurons in some of the central catecholamine cell groups and provide a morphological basis for interactions between the peptide and amines.


Expression of Notch signaling components in cutaneous foreign body and sarcoidal granulomas and fusing macrophages.
The evolutionarily conserved Notch signaling pathway affects tissue-specific cell differentiation, proliferation, and apoptosis. In the immune system, Notch has been implicated in the development and function of both adoptive and innate immune cells. Notch signaling is initiated by Notch receptor binding to cognate ligands, which results in the enzymatic cleavage and intranuclear translocation of the intracellular domain of Notch receptor (ICN). Recent murine models of chronic inflammation highlighted a critical role for a Notch ligand, Delta-like ligand (Dll)-4, in granuloma formation. In this study, we aimed to assess Notch-1 receptor activation and Dll4 expression in human cutaneous granulomas and in cultured human macrophages and multinucleated giant cells. ICN1 and Dll4 expression was evaluated by immunohistochemistry of cutaneous foreign body (n = 15) and sarcoidal (n = 19) granulomas. The results showed consistent intranuclear staining for ICN1 in foreign body but not in sarcoidal granulomas and strong cytoplasmic staining for Dll4 in mononuclear histiocytes and multinucleate giant cells in both types of granulomas. Additionally, immunofluorescence confocal microscopy showed ICN1 and Dll4 expression by cultured human macrophages undergoing fusion in the presence of granulocyte-macrophage colony-stimulating factor and interleukin-4. These findings indicate a potential role for the Notch-1-Dll4 signaling pathway in foreign body-induced granulomatous reactions and possibly distinct Notch pathway utilization in sarcoidal granulomas.


Analysis of new classes of recreational drugs in sewage: synthetic cannabinoids and amphetamine-like substances.
The analysis of sewage for the residues of commonly used illicit drugs has successfully been applied as a suitable approach for estimating community illicit drug use. The drug market is increasingly dynamic with new substances continually being marketed for recreational purposes. In this study, ultra-high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to simultaneously and quantitatively detect the exogenous biomarkers of new classes of recreational drugs in sewage collected from three different Norwegian cities (Oslo, Bergen, Hamar). The samples were screened for the presence of khat (d-norpseudoephedrine and cathinone), mephedrone, pseudoephedrine, 7-aminoflunitrazepam, para-methoxyamphetamine (PMA), para-methoxy-N-methylamphetamine (PMMA) and a selection of urinary metabolites of synthetic cannabinoids collectively termed ´Spice´ (5-3-1-naphthoyl-1H-indol-1-yl-pentanoic acid (JWH 018 N-pentanoic acid), 1-5-hydroxypentyl-1H-indol-3-ylnaphthalen-1-yl-methanone (JWH 018 N-5-hydroxypentyl), 4-3-1-naphthoyl-1H-indol-1-yl-butanoic acid (JWH 073 N-butanoic acid), 1-4-hydroxybutyl-1H-indol-3-ylnaphthalen-1-yl-methanone (JWH 073 N-4-hydroxybutyl), 1-5-hydroxypentyl-1H-indol-3-yl4-methylnaphthalen-1-yl-methanone (JWH 122 N-5-hydroxypentyl), 1-5-fluoro-4-hydroxypentyl-1H-indol-3-ylnaphthalen-1-ylmethanone (AM2201 N-4-hydroxypentyl), and 1-5-hydroxypentyl-1H-indol-3-yl4-methoxyphenyl-methanone (RCS-4 N-5-hydroxypentyl)). Limits of detection were 1 ng/L for amphetamine like compounds and 5 ng/L for the metabolites of synthetic cannabinoids while the limits of quantification were 3 and 15 ng/L, respectively. Three of the fourteen selected biomarkers (cathine, pseudoephedrine and the synthetic cannabinoid metabolite JWH-018 N-5-hydroxypentyl) were detected in sewage, alongside the illicit drugs (and/or metabolites) typically found in sewage (cocaine, benzoylecognine, methamphetamine, MDMA, and THC-COOH). The khat biomarker d-norpseudoephedrine was detected in Oslo sewage at a mean concentration of 93 ng/L that represents a daily load of 54 mg/day/1000 inhabitants. Pseudoephedrine was present at mean concentrations of between 27 and 67 ng/L representing normalized daily loads of between 10 (Hamar) and 24 mg/day/1000 inhabitants (Bergen). The daily normalized loads of JWH-018 N-5-hydroxypentyl were between 49 (Oslo) and 62 mg/day/1000 inhabitants (Hamar). This study demonstrates for the first time that sewage biomarker analysis can be applied to evaluate not only the use the traditional illicit drugs (cocaine, cannabis and amphetamines), but also the use of certain new synthetic drugs.


Psychogeriatric Services at Certified Home Health Agencies: Case Reports and Guidelines for Psychiatric Consultants.
Because of the unmet mental health needs of older persons in the community, Medicare-certified home health agencies are increasingly taking the role of health providers. Here the authors review their experience and argue that the pathology seen in home mental health care situations is similar to that seen by specialized mental health outreach teams. Also the relations between the home care team and the psychiatric consultant require skillful management even when the team are mental health specialists. The authors offer guidelines for psychiatric consultants, given the extent to which home care services survive in a volatile, cost-contained environment.


Reflectance confocal microscopy analysis of equivocal melanocytic lesions with severe regression.
The differential diagnosis between regressing nevi and melanoma might be challenging; regressing areas can represent a confounding factor for the diagnosis and the histology still remain mandatory to rule out melanoma. Reflectance confocal microscopy may add valuable information by revealing features suggestive of the nature of the melanocytic proliferation. To assess the impact of confocal microscopy in the management of regressive melanocytic lesions. The dermoscopic analysis of 92 melanocytic lesions showing that more than 30% of regressions have been retrospectively considered, among them, 32 melanocytic lesions with a 7 check point list ≥3 they were assessed at the rcm and subsequently excised. For each selected lesion, dermoscopic features of regression (white scar-like areas, blue areas, blue white areas), distribution of regressing areas (central, peripheral, or both) and the percentage of regression have been examined by an expert in dermoscopy, blinded to the histological and confocal diagnosis. Subsequently, two experts in confocal microscopy revaluated, blinded from histology, RCM images. Of the 32 lesions analyzed, 23 (71.5%) were diagnosed histologically as nevi, and 9 (28.5%) as melanomas. 26 of 32 lesions (81.5%) exhibited regression >50% of the overall. On RCM, 11 lesions have been interpreted as malignant and 21 as benign. On RCM the majority of nevi exhibited regular architecture without cytological atypia. Epidermal disarray, pagetoid infiltration, disarranged dermo-epidermal junction architecture and atypical nests were considered as suspicious for malignancy. Good concordance between confocal readers has been detected. A combined dermoscopic/confocal approach can be used for the management of lesions exhibiting dermoscopic features of regression in order to provide a more conclusive pre-histological diagnosis avoiding a high number of unnecessary excisions. Limits of this study were represented by the relatively small number of lesions and the retrospective approach. Further, prospective studies on a larger number of cases, will be necessary in order to compare the efficacy of dermoscopy alone versus dermoscopy in combination with RCM for the evaluation of regression, suspected pigmented lesions.


Periphery of porcine hepatic lobes has the smallest length density of hepatic sinusoids and bile canaliculi: A stereological histological study with implications for liver biopsies.
Porcine liver is widely used in hepatologic research as a large animal model with many anatomical and physiological similarities with humans. However, only limited information on porcine liver spatial microstructure has been published, especially regarding the hepatic sinusoids and bile canaliculi. The aim of our study was to quantify the sinusoidal and bile canalicular network in healthy male and female porcine livers and to map the variability of these structures with heterogenous distribution to improve the evaluability of liver biopsy samples. Livers from 12 healthy piglets (6 females and 6 neutered males) were sampled into 36 tissue samples per organ, representing six hepatic lobes and three different regions related to the hepatic vasculature (peripheral, paracaval and paraportal region). Histological sections were processed with a random orientation of the cutting plane. The endothelium and the bile canaliculi were stained using Ricinus communis agglutinin I lectin histochemistry. The length densities of hepatic sinusoids LV(sinusoids,liver), of bile canaliculi LV(bile canaliculi,liver) and volume fraction VV(sinusoids,liver) and surface density SV(sinusoids,liver) of sinusoids were estimated using stereological methods. The newly acquired morphometric data were compared with previously published data on density of porcine hepatocytes and fractions of connective tissue. The peripheral region had smallest LV(sinusoids,liver), smallest LV(bile canaliculi,liver) and greatest VV(sinusoids,liver). The six hepatic lobes had statistically comparable length densities of both sinusoids and bile canaliculi, but the left lateral lobe had smallest VV(sinusoids,liver). Regions with greater LV(sinusoids,liver) had also greater LV(bile canaliculi,liver) and SV(sinusoids,liver) and were accompanied by greater density of smaller hepatocytes. Regions with smaller LV(sinusoids,liver) and LV(bile canaliculi,liver) contained a greater fraction of interlobular connective tissue. The length density of hepatic sinusoids is smaller in the peripheral regions of the porcine liver than in other regions related to the hepatic vasculature - paracaval and paraportal regions, and smaller in castrated males than in females. Greater length density of liver sinusoids was linked with greater local density of bile canaliculi, with local increase in the density of smaller hepatocytes and, simultaneously, with smaller fractions of hepatic connective tissue. The intrahepatic and inter-sexual variability of the porcine liver morphology needs to be taken into account when designing and interpreting experiments involving the histological quantification of the microvascular network. The complete primary morphometric data describing the distribution of morphometric parameters within porcine liver were made available in a form facilitating the power analysis to justify the minimal number of tissue samples or animals required when designing further histological evaluation studies. The macroscopic map of microvessels and bile canaliculi variability facilitates their assessment in liver biopsies in the pig.


Acenaphthoimidazolylidene-Ligated Palladacycle Enabled Suzuki-Miyaura Cross-Coupling Employing Equimolar Organoboron for Tri-Ortho-Substituted Bi(hetero)aryls and Teraryls.
The Pd-catalyzed Suzuki-Miyaura cross-couplings (SMRs) are utilized as the most practical method to construct C-C bond, especial for biaryls. However, a major disadvantage of current protocols is the requirement of excess organoboron coupling partner (1.5-3.0 equiv.). Herein, a novel palladacyclic 1,3-bis(2,6-diisopropylphenyl)acenaphthoimidazol-2-ylidene (AnIPr) precatalyst possessing a chiral oxazoline was designed, which enabled a general protocol towards bulky tri-ortho-substituted biaryls, ternaphthalenes and diarylanthracenes via the Pd-catalyzed SMR employing equimolar organoborons and aryl bromides. A remarkable scope of substrates with various functional groups and heterocycles were well compatible with an adaptability to synthesize useful ligands.


Transcranial Direct Current Stimulation of the Temporoparietal Junction and Inferior Frontal Cortex Improves Imitation-Inhibition and Perspective-Taking with no Effect on the Autism-Spectrum Quotient Score.
Lesions to brain regions such as the temporoparietal junction (TPJ) and inferior frontal cortex (IFC) are thought to cause autism-spectrum disorder (ASD). Previous studies indicated that transcranial direct current stimulation (tDCS) of the right TPJ improves social cognitive functions such as imitation-inhibition and perspective-taking. Although previous work shows that tDCS of the right IFC improves imitation-inhibition, its effects on perspective-taking have yet to be determined. In addition, the role of the TPJ and IFC in determining the Autism-Spectrum Quotient (AQ), which is a measure of autism spectrum traits, is still unclear. Thus, the current study performed tDCS on the right TPJ and the right IFC of healthy adults, and examined its effects on imitation-inhibition, perspective-taking and AQ scores. Based on previous studies, we hypothesized that anodal tDCS of the right IFC and right TPJ would improve imitation-inhibition, perspective-taking and the AQ score. Anodal tDCS of the right TPJ or IFC significantly decreased the interference effect in an imitation-inhibition task and the cost of perspective-taking in a perspective-taking task, in comparison to the sham stimulation control. These findings indicated that both the TPJ and the IFC play a role in imitation-inhibition and perspective-taking, i.e., control of self and other representations. However, anodal stimulation of the right TPJ and the right IFC did not alter participants' AQ. This finding conflicts with results from previous brain imaging studies, which could be attributed to methodological differences such as variation in sex, age and ASD. Therefore, further research is necessary to determine the relationship between the TPJ and IFC, and the AQ.


Longitudinal CT and MRI Characteristics of Unilocular Thymic Cysts.
Background Isoattenuating and hyperattenuating thymic cysts at CT are often misinterpreted as lymphadenopathy or thymic epithelial neoplasms. Purpose To evaluate the longitudinal change in thymic cyst appearance at CT and MRI. Materials and Methods All chest MRI studies showing thymic cysts between July 2008 and December 2019, identified from a retrospective search of a quality assurance database, were included in this study if initial CT depicted a thymic lesion, the patient was referred for follow-up MRI for characterization, and the baseline (ie, index) MRI indicated a cystic lesion. Follow-up CT scans and/or MRI scans were identified through July 2020. Thymic cyst characteristics, such as size, location, and morphologic features, as well as CT and MRI characteristics, were recorded. Change in size, attenuation, and T1-weighted MRI signal was assessed longitudinally. Descriptive statistics of longitudinal change were tabulated. Results A total of 244 chest MRI studies in 140 patients with 142 unique cysts and 392 CT examinations (636 total examinations and 645 thymic cysts-nine examinations with two cysts each) were evaluated. The median follow-up duration was 2.2 years. Thirty-three patients with 34 unique cysts (34 of 142 cysts [24%]) underwent imaging follow-up for more than 5 years. Thymic cysts followed up for more than 5 years were most commonly saccular (189 of 274 cysts [69% axially]) and retrosternal (14 of 34 cysts [41%]). Craniocaudal dimension was larger than transverse and anteroposterior dimensions in 223 of 274 cysts (81%). Mean thymic cyst attenuation was 25 HU (range, 15-100 HU). Five of 31 cysts (16%) exhibited wall calcification. The median cyst wall thickness was 2.0 mm (range, 0.9-3.0 mm). Most thymic cysts changed in volume (31 of 34 cysts [91%]), CT attenuation (15 of 35 cysts [43%]), and T1-weighted MRI signal (12 of 18 cysts [67%]) over time. None developed mural irregularity, nodularity, or septations. Conclusion Unilocular thymic cysts, defined at index MRI, never developed irregular wall thickening, mural nodularity, or septations that would raise concern for malignant transformation. However, these cysts showed mural calcification and change in size, CT attenuation, and MRI signal over more than 5 years of follow-up. © RSNA, 2021 Online supplemental material is available for this article.


Polar distribution dynamics of Con A binding sites in embryo sacs is temporally coupled by the fertilization process.
The binding site distribution of concanavalin agglutinin (Con A) and wheat germ agglutinin (WGA) on embryo sacs at various developmental stages of Torenia fournieri L was studied by using a cooled Charge Coupled Device (CCD) and fluorescent Con A and WGA probes. The distribution patterns of Con A and WGA binding sites on embryo sacs changed during the fertilization process. The fluorescent signal indicating Con A binding sites was distributed evenly on the surface of the embryo sac wall before anthesis, was much denser on the micropylar end of the embryo sac wall and looked like a corona on the day of anthesis. After pollination, stronger fluorescence was present on the micropylar end of the embryo sac wall and the filiform apparatus (FA), showing an obvious polar distribution. When the pollen tube entered the embryo sac and reached a synergid, the fluorescence was still concentrated on the micropylar end and FA, and started to appear on the synergid. After fertilization, the polar distribution of the fluorescence gradually disappeared and an even distribution pattern was observed again on the embryo sac wall. These results revealed that the dynamic distribution of Con A binding sites was temporally coupled with the process of fertilization. WGA binding site distribution on the embryo sac was also investigated and showed a simple pattern but also regularly changed during the process of fertilization. The variation of these lectin binding sites during the fertilization process suggests that lectin binding site interactions may play a role in the process.


Investigation of Masked N-Acyl-N-isocyanates: Support for Oxadiazolones as Blocked N-Isocyanate Precursors.
In contrast to carbon-substituted isocyanates that are common building blocks, N-substituted isocyanates remain underdeveloped and reports on their N-acyl derivatives (i. e. amido-isocyanates) are exceedingly rare. Herein, amido-isocyanates were investigated in the context of syntheses of aza-tripeptide and hydantoins subunits starting from simple bench-stable precursors. A key finding is that the amido-isocyanate formed in situ cyclized to yield an oxadiazolone, and that under suitable reaction conditions this heterocycle is a traceless blocked (masked) N-isocyanate. Using organic bases as catalysts and upon heating, oxadiazolone formation is observed, and various nucleophiles to provide the desired aza-dipeptides or hydantoins in moderate to high yields. Further support for an amido-isocyanate intermediate was obtained using carboxylic acids as nucleophiles, affording N-acylhydrazide products.


Ranitidine reduces ischemia/reperfusion-induced liver injury in rats by inhibiting neutrophil activation.
We previously reported that ranitidine, an H(2) receptor antagonist, inhibited neutrophil activation in vitro and in vivo, contributing to reduce stress-induced gastric mucosal injury in rats. In this study, we examined whether ranitidine would reduce ischemia/reperfusion-induced liver injury, in which activated neutrophils are critically involved, in rats. We also examined the effect of famotidine, another H(2) receptor antagonist, on leukocyte activation in vitro and after ischemia/reperfusion-induced liver injury in rats to know whether inhibition of neutrophil activation by ranitidine might be dependent on its blockade of H(2) receptors. Ranitidine inhibited the activation of neutrophils in vitro as reported previously, whereas famotidine significantly enhanced it. Ranitidine inhibited the production of tumor necrosis factor-alpha (TNF-alpha) in monocytes stimulated with lipopolysaccharide in vitro, whereas famotidine did not. Although hepatic ischemia/reperfusion-induced increases in hepatic tissue levels of TNF-alpha, cytokine-induced neutrophil chemoattractant, and hepatic accumulation of neutrophils were inhibited by intravenously administered 30 mg/kg ranitidine, these increases were significantly enhanced by 5 mg/kg i.v. famotidine. The decreases in both hepatic tissue blood flow and bile secretion and the increases in serum levels of transaminases seen after reperfusion were significantly inhibited by ranitidine, whereas these changes were more marked in animals given famotidine than in controls. These observations strongly suggested that ranitidine could reduce ischemia/reperfusion-induced liver injury by inhibiting neutrophil activation directly, or indirectly by inhibiting the production of TNF-alpha, which is a potent activator of neutrophils. Furthermore, the therapeutic efficacy of ranitidine might not be explained solely by its blockade of H(2) receptor.


Synthesis of 3 10-helix-inducing constrained analogues of L-proline.
A bicyclic indolizidinone carboxylic acid and a tricyclic constrained analogue of l-proline were synthesized and evaluated for their ability to induce helix formation as l-Ala tetrapeptides. Variable-temperature NMR, DMSO titration, CD spectra, and X-ray structure analyses, in conjunction with molecular modeling, confirmed the existence of 3(10)-helical motifs with di- and tetrapeptides of l-Ala.


Posterior soft tissue repair in total hip arthroplasty: a randomized controlled trial.
Posterior soft tissue repair is a well-known procedure in total hip arthroplasty (THA). Many reports have shown the advantage of posterior soft tissue repair in reducing the dislocation rate; however, we were unable to find any randomized trials in the literature. This article describes a randomized trial performed to investigate the effect of posterior soft tissue repair on the dislocation rate after 298 THAs in 291 patients. Preoperatively, patients were randomized into 2 groups: posterior soft tissue repair after insertion of the components, or no posterior soft tissue repair. One year postoperatively, 12 patients had died and 10 were lost to follow-up, leaving 276 THAs in 265 patients for analysis. A posterior soft tissue repair had been performed in 134 and no repair in 141 THA. One year postoperatively, 3 dislocations (2%) had occurred in the repaired group and 7 in the unrepaired group (5%); the difference was not significant (P=.3). There were 2 sciatic nerve palsies in the repaired group, 1 of which was directly related to the posterior soft tissue repair.In this randomized, controlled trial of 276 THAs followed for 1 year, there was a tendency for a lower dislocation rate, although not statistically significant, when a posterior repair was performed. If reattaching the posterior tendons, it should be performed with caution with respect to the sciatic nerve.


Renal oncocytoma: angiographic features of two cases.
Two cases of renal oncocytoma (proximal tubular adenoma) with a similar angiographic appearance are reported. The vascular supply to the lesion tends to be arranged in "spoke-wheel" pattern with vessels radiating toward the center of the lesion; the angiographic appearance may help to differentiate renal oncocytoma from hypernephroma preoperatively.


The effects of ephedrine on maternal hypothermia in caesarean sections: a double blind randomized clinical trial.
The purpose of the study was to investigate the effect of bolus and the combination of bolus and infusion of ephedrine on maternal hypothermia which are used for treating maternal hypotension under spinal anaesthesia. 110 ASA I-II patients who developed maternal hypotension were included into the study. Spinal anaesthesia was performed with 12.5 mg heavy bupivacaine + 15 µg fentanyl. Group I: Ephedrine bolus 5 mg plus ephedrine infusion, Group B: Ephedrine bolus 5 mg plus normal saline infusion. The systolic blood pressure was allowed to range between 20% from baseline values. Ephedrine solution infusion started after hypotension occurred (0.5 mg/minute). The body temperature under 35.5°C was accepted as hypothermia. The newborns' rectal temperature was measured. Moreover, the Apgar scores, umbilical vein-arterial blood gas and acid-base status were evaluated. In Group I, the body core temperatures which were measured at 9, 18, 33, and 39th minutes were significantly higher than Group B (p < 0.05). The prevalence of maternal hypothermia in Group I was significantly lower than the Group B, which were as 65.5% (36/55) and 85.5% (47/55), respectively (p < 0.05). In Group I, the newborn rectal temperatures and the total dose of ephedrine were significantly higher than Group B (p < 0.05). In Group I, the systolic and mean blood pressures were higher than Group B (p < 0.05). As a result, we found that combined bolus and infusion of ephedrine for treating maternal hypotension prevents maternal and neonatal hypothermia during caesarean section under spinal anaesthesia compared to bolus administrations alone.


Detection of multiple viral and bacterial infections in acute exacerbation of chronic obstructive pulmonary disease: a pilot prospective study.
Few studies have evaluated the contribution of multiple virus and bacterial infections in acute exacerbation of chronic obstructive pulmonary disease. This study estimated the burden of multiple viral and bacterial respiratory infections in moderate to very severe chronic obstructive pulmonary disease patients that were prospectively followed-up during a 12-month pilot study. Clinical data were collected monthly and sputum was collected at the time of each acute exacerbation event. Classical culture techniques for bacteria and multiplex polymerase chain reaction (PCR) and microarray detection assays were performed to identify viral and atypical bacterial pathogens in the sputum. Overall, 51 patients were included and 45 acute exacerbation events were investigated clinically and microbiologically. Among the 45 acute exacerbation events, 44% had evidence of viral infection involving human rhinovirus (HRV) and metapneumovirus (hMPV) in 20% and 18%, respectively. Intracellular bacteria were not found in sputum by PCR. Common bacterial pathogens were identified in 42% of acute exacerbation patients, most frequently Branhamella catarrhalis, Streptococcus pneumoniae and Haemophilus influenzae. Viral or virus and bacteria co-infections were detected in 27% of acute exacerbation events (n = 12) with HRV and hMPV involved in 92% of cases. Patients with co-infections did not present greater clinical severity scores at exacerbation and more recurrence of acute exacerbation events at 3 and 6 months than those with single infections (P > 0.4). These results suggest that HRV and hMPV may be contributors or cofactors of AECOPD. These findings indicate that viral or virus and bacterial co-infections do not impact significantly on the clinical severity of acute exacerbation of chronic obstructive pulmonary disease and recurrence at 3 and 6 months.


Short-duration, high-dose urokinase infusion for recanalization of occluded saphenous aortocoronary bypass grafts.
Thrombolytic recanalization of arterial bypass grafts has been pursued aggressively in the peripheral circulation but not in the coronary circulation. In an attempt to apply peripheral transcatheter thrombolytic techniques to the coronary circulation, nine patients with 10 occluded saphenous aortocoronary bypass grafts underwent recanalization procedures using a short-duration, high-dose urokinase infusion. Urokinase was infused at the occluded graft orifice at a rate of 600 units/min. The average infusion time was 1 hr, 26 min. The average urokinase dose was 435,000 units. Graft recanalization was achieved in eight (80%) of 10 grafts, although only six (60%) of 10 grafts were widely patent at the end of the procedure. All successfully recanalized grafts required balloon angioplasty of underlying stenoses. No complications, specifically myocardial infarction or cerebrovascular accident, were encountered. We have shown that occluded aortocoronary bypass grafts can be recanalized successfully by using a short-duration, high-dose urokinase infusion. It appears that, with attention given to angiographic techniques that minimize clot manipulation, recanalization can be accomplished safely in a majority of cases.


Modified Mainz pouch II (Sigma Rectum pouch) urinary diversion: 12 years experience.
To analyze the outcome of Mainz Pouch II urinary diversion related to complications and life quality. From 1995 to 2006, a total of 67 patients (60 male and 7 female, mean age 58.4 years, range 48 to 70) who underwent modified ureterosigmoidostomy (Mainz Pouch II) procedure have been retrospectively analyzed. The mean follow-up was 18 (1 - 72) months and it was available for 56 patients (84%). Early and late postoperative complications as well as quality of life after surgery were analyzed. An clinical questionairre has been used for examination specific urinary diversion items. Early postoperative complications (<30 days) we-re detected in 9 patients (13%) and late complications (>30 days) in 19 patients (28%). Early complications consisted of urine leakage of moderate degree in 5 (7%) and ileus requiring surgical revision in 4 (6%) patients. The late complications included acute pyelonephritis in 12 patients (18%) and uretero-sigmoidal anastomotic site stenosis in 7 (11%). Ureterosigmoidal anastomotic site stenosis was detected in 7 patients with 7 renoureteric units (RU). In 4 RU, metal Strecker stent was successfully applied. In 3 RU, permanent nephrostomy catheter was applied. Oral alkalizing agents were applied in 22 patients (33%) due to metabolic acidosis. Two patients died due to severe acidosis. Continent rate was 96%. The mean voiding frequency during the day and night was 5.2+/-1.8 and 2.7+/-0.5, respectively. Global life quality was bad for 3 (8%), acceptable for 15 (38%) and good for 22 (54%) patients. The Mainz Pouch II urinary diversion is simple and safe procedure regarding complications rate, continence and quality of life. It is good alternative to other forms of continent urinary diversion. Patient selection and compliance following by meticulous follow-up are of utmost importance for successful operative outcome.


Real-time cdPCR opens a window into events occurring in the first few PCR amplification cycles.
Polymerase chain reaction (PCR) end-point limiting dilution techniques, collectively termed "digital PCR (dPCR)", have been proposed as providing a potentially primary method for DNA quantification. We are evaluating several commercially available dPCR systems for use in certifying mass concentration in human genomic DNA reference materials. To better understand observed anomalies among results from chamber- and droplet-dPCR (cdPCR and ddPCR) systems, we have developed a graphical tool for evaluating and documenting the performance of PCR assays in real-time cdPCR systems: the ogive plot, the cumulative distribution of crossing threshold values. The ogive structure appears to embed information about early amplification events. We have successfully simulated ogives observed with different assays and reaction conditions using a four-stage amplification model parameterized by the probability of creating an intact 1) first generation "long" amplicon of indeterminate length from an original DNA target, 2) second generation defined-length amplicon from a long amplicon, and 3) defined-length amplicon from another defined-length amplicon. We are using insights from this model to optimize dPCR assay design and reaction conditions and to help validate assays proposed for use in value-assigning DNA reference materials.


Isolated subtalar fusion for neglected painful intra-articular calcaneal fractures.
The objective of this study was to evaluate the radiological and clinical outcomes of treatment of subtalar arthrodesis in patients developing talocalcaneal arthrosis secondary to intra-articular calcaneal fractures. The study included 20 patients (21 feet) who underwent subtalar arthrodesis due to symptomatic subtalar arthrosis following conservative treatment for intra-articular calcaneal fracture between 2005 and 2011. Autograft or allograft was used in 11 patients. Patients were evaluated clinically using the American Orthopedic Foot and Ankle Society (AOFAS) ankle hindfoot score. Hindfoot alignment, quality of subtalar fusion and arthritis occurring in other joints were used for the radiological evaluations. Mean duration of follow-up was 43 (range: 21 to 83) months. Mean preoperative AOFAS score was 61.7 (range: 40 to 67) and mean postoperative AOFAS score was 84.2 (range: 65 to 94). The difference between scores was statistically significant (p=0.001). Six patients had excellent, 8 good and 6 fair results. Complete fusion was achieved in 19 patients (20 feet). In 2 patients, arthritic changes were radiologically observed in the midtarsal joints. These changes were not symptomatic. There were no statistically significant differences between pre- and postoperative radiological measurements. No patients experienced malunion. While subtalar arthrodesis appears to provide radiological and clinical benefits, it may cause moderate and asymptomatic osteoarthritis in the midtarsal joints.


Balanced Ambipolar Poly(diketopyrrolopyrrole-alt-tetrafluorobenzene) Semiconducting Polymers Synthesized via Direct Arylation Polymerization.
The synthesis of an ambipolar π-conjugated copolymer consisting of alternating diketopyrrolopyrrole and tetrafluorobenzene via direct arylation polymerization (DAP) is reported. Two different combinations of monomers are investigated under various catalytic conditions for DAP. The target polymer obtained under an optimized catalytic condition shows minimal structural defects, a number-average molecular weight of 33.2 kDa, and balanced electron and hole mobility of 1 × 10(-2) cm(2) V(-1) S(-1) in the organic field-effect transistors fabricated and tested under ambient conditions.


Maternal serotonin is crucial for murine embryonic development.
The early appearance of serotonin and its receptors during prenatal development, together with the many effects serotonin exerts during CNS morphogenesis, strongly suggest that serotonin influences the development and maturation of the mammalian brain before it becomes a neuromodulator/neurotransmitter. Sites of early serotonin biosynthesis, however, have not been detected in mouse embryos or extraembryonic structures, suggesting that the main source of serotonin could be of maternal origin. This hypothesis was tested by using knockout mice lacking the tph1 gene, which is responsible for the synthesis of peripheral serotonin. Genetic crosses were performed to compare the phenotype of pups born from homozygous and heterozygous mothers. Observations provide the first clear evidence that (i) maternal serotonin is involved in the control of morphogenesis during developmental stages that precede the appearance of serotonergic neurons and (ii) serotonin is critical for normal murine development. Most strikingly, the phenotype of tph1-/- embryos depends more on the maternal genotype than on that of the concepti. Consideration of the maternal genotype may thus help to clarify the influence of other genes in complex diseases, such as mental illness.


Changing drug choice through conditioned reinforcement: comment on Alessi, Roll, Reilly, and Johanson (2002).
The study by S. M. Alessi, J. M. Roll, M. P. Reilly, and C.-E. Johanson (2002) showed that in human volunteers a choice of placebo over diazepam could be reversed by pairing the drug with a higher monetary reward and the placebo with a lower monetary reward for performance on a computer-based task. The results provide a clear laboratory model of how initially neutral or aversive effects of a drug can be altered by association with positive reinforcement to establish the drug as a conditioned reinforcer and to generate drug-seeking behavior. Parallels from the animal and human literatures are discussed, as are potential future studies and applications of this approach for preventing and reducing drug abuse in humans.


Structure-function analyses of a PL24 family ulvan lyase reveal key features and suggest its catalytic mechanism.
Ulvan is a major cell wall component of green algae of the genus Ulva, and some marine bacteria encode enzymes that can degrade this polysaccharide. The first ulvan-degrading lyases have been recently characterized, and several putative ulvan lyases have been recombinantly expressed, confirmed as ulvan lyases, and partially characterized. Two families of ulvan-degrading lyases, PL24 and PL25, have recently been established. The PL24 lyase LOR_107 from the bacterial Alteromonadales sp. strain LOR degrades ulvan endolytically, cleaving the bond at the C4 of a glucuronic acid. However, the mechanism and LOR_107 structural features involved are unknown. We present here the crystal structure of LOR_107, representing the first PL24 family structure. We found that LOR_107 adopts a seven-bladed β-propeller fold with a deep canyon on one side of the protein. Comparative sequence analysis revealed a cluster of conserved residues within this canyon, and site-directed mutagenesis disclosed several residues essential for catalysis. We also found that LOR_107 uses the His/Tyr catalytic mechanism, common to several PL families. We captured a tetrasaccharide substrate in the structures of two inactive mutants, which indicated a two-step binding event, with the first substrate interaction near the top of the canyon coordinated by Arg320, followed by sliding of the substrate into the canyon toward the active-site residues. Surprisingly, the LOR_107 structure was very similar to that of the PL25 family PLSV_3936, despite only ∼14% sequence identity between the two enzymes. On the basis of our structural and mutational analyses, we propose a catalytic mechanism for LOR_107 that differs from the typical His/Tyr mechanism.


Antigenic mimicry of natural L-peptides with retro-inverso-peptidomimetics.
Three analogues of the model peptide of sequence IRGERA corresponding to the COOH-terminal residues 130-135 of histone H3 were synthesized, and their antigenicity, immunogenicity, and resistance to trypsin were compared to those of the natural L-peptide. The three analogues correspond to the D-enantiomer, containing only D-residues, and two retro-peptides containing NH-CO bonds instead of natural peptide bonds. The chirality of each residue was maintained in the retro-peptide and inverted in the retro-inverso-peptide. Antibodies to the four peptide analogues were produced by injecting BALB/c mice with peptides covalently coupled to small unilamellar liposomes containing monophosphoryl lipid A. Each of the four peptide analogues induced IgG antibodies of various subclasses. The IgG3 antibodies reacted similarly with the four analogues, whereas antibodies of the IgG1, IgG2a, and IgG2b isotypes showed strong conformational preferences for certain peptides. The retro-inverso-peptide IRGERA mimicked the structure and antigenic activity of the natural L-peptide but not of the D- and retro-peptides, whereas the retro-peptide IRGERA mimicked the D-peptide but not the L- and retro-inverso-peptides. The equilibrium affinity constants (Ka) of three monoclonal antibodies generated against the L- and D-peptides with respect to the four peptide analogues were measured in a biosensor system. Large differences in Ka values were observed when each monoclonal antibody was tested with respect to the four peptides. The use of retro-inverso-peptides to replace natural L-peptides is likely to find many applications in immunodiagnosis and as potential synthetic vaccines.


'Is Grandma Still There?' A Pastoral and Ethical Reflection on the Soul and Continuing Self-identity in Deeply Forgetful People.
This metaphysical and pastoral reflection focuses on a question that over several decades has been posed to me by many family caregivers for deeply forgetful people (persons with dementia). The question may take different forms: Is my loved one still there underneath all of this confusion? Is my loved one's soul still present? Will she come to rest fully intact in the arms of a Supreme Being? Could she be 'gone' but already somewhere experiencing the fullness of divine love? This reflection provides a pastoral response to this big question that has to do with the need to find meaning in caring for deeply forgetful people.