Fatigue in cerebral palsy: a critical review.
Fatigue contributes to the deterioration or cessation of walking ability in adults with cerebral palsy (CP). However, conflict exists as to its role. Studies involving functional tasks reported increased, and earlier onset of, fatigue in CP, whereas laboratory studies have reported individuals with CP to be more fatigue-resistant than their peers. A critical review of the literature related to fatigue in CP was conducted. This review describes factors that contribute to the observed fatigue resistance in laboratory tasks and how a decreased force-production in CP can result in higher energy expenditure to perform the same amount of work as their peers. More research regarding the process of fatigue and recovery for individuals with CP is needed; specifically studies that focus on functional movements requiring the integration of the whole body, thereby stressing the neuromuscular system in a different way than previously explored.


Worsening Hypoxia Post Lung Transplant: What has Changed?
Platypnoea-orthodeoxia is a rare, but under-diagnosed clinical entity, characterised by postural hypoxia and breathlessness. Underlying pathology is inter-atrial shunt or pulmonary vascular malformation, but what anatomical distortion / physiological alterations initiates right to left shunt, usually against the pressure gradient remains unknown.


Allometry of the quasi-pipe (qPipe) model for estimating tree leaf area and tree leaf mass applied to plant functional types.
The allometry of the pipe model quantifies the approximate proportionality between the tree leaf amount and the stem cross-sectional area at the crown base (ACB). It is useful for estimating and modeling carbon fixation abilities of trees but requires climbing the tree and is thus unsuitable for large-scale studies. Here, we adopted a previously proposed allometry (hereafter the quasi-pipe (qPipe) model allometry) formulating the relationship between the tree leaf amount and a surrogate of ACB, ACB_Est, calculated from tree dimensions measurable from the ground. Using published/unpublished data for 962 trees of 159 species collected between tropical rainforests and boreal forests, we established pipe and qPipe model allometries for evergreen-conifer, deciduous-conifer, evergreen-broadleaf, and deciduous-broadleaf plant functional types (PFTs). For the leaf area per tree (LA), allometric lines on a log-log plane were almost identical among the four PFTs in both models, with slopes of ~ 1. For the leaf mass per tree (LM), however, the allometric lines separated among the four PFTs in both models and had slopes greater than 1, indicating that the proportionality assumed in the pipe model held for LA but not LM. The applicability of the qPipe model in estimating the stand-scale leaf amount was further examined.


Catalytic analysis of a recombinant D-hydantoinase from Agrobacterium tumefaciens.
The D-hydantoinase gene of a wild strain of Agrobacterium tumefaciens BQL9 had 99.78% nucleotide sequence identity with other available Agrobacterium genes. The resulting amino acid sequence showed two important substitutions affecting two alpha-helixes in the secondary structure of the protein. The union of Mn2+ to the protein was essential for activating the enzyme and was independent of the temperature. D-Hydantoinase only was inactivated in the presence of 70 mM EDTA and at over 40 degrees C. The enzyme showed both hydantoinase and pyrimidinase activities, but only with the D-enantiomers of the substrates. Activity was greater for substrates with apolar groups in the number 5 carbon atom of the hydantoin. The native structure of the N-terminal end of this D-hydantoinase proved to be indispensable to its enzymatic activity.


Antiplasmodial activity of Mezoneuron benthamianum leaves and identification of its active constituents.
Decoctions of the leaves of M. benthamianum Baill. are used by traditional healers in Guinea to treat malaria and this use was validated by a preliminary clinical assay. To evaluate the in vitro antiplasmodial activity and to identify active compounds from extracts of M. benthamianum leaves. Antiplasmodial activity of extracts, fractions and pure compounds was evaluated in vitro against a chloroquine-sensitive strain of Plasmodium falciparum (3D7) using the measurement of the plasmodial lactate dehydrogenase activity. Selectivity of extracts and purified compounds for Plasmodium parasites was evaluated by using WST-1 test on HeLa human cells. Compounds were isolated using normal phase silica gel column chromatography and prepHPLC and their structures elucidated using extensive spectroscopic analysis. Hydroethanolic extracts (70% v/v) of M. benthamianum leaves showed a moderate in vitro activity against P. falciparum 3D7, with IC50 in the range 22.5 - 32.6µg/mL, depending on the batch; while a dark precipitate formed during ethanol evaporation showed higher activity (IC50 =6.5µg/mL). The fractionation was performed on this most active fraction and was followed by in vitro antiplasmodial assay. Active compounds (5, 7, 8) belong to several phytochemical classes, contributing together to the global antiplasmodial activity of the hydroethanolic extract against P. falciparum parasite. This study finally allowed the isolation of three diterpenes including two new compounds named Mezobenthamic acids A (1) and B (2) and neocaesalpin H (3), as well as quercetin (4), kaempferol (7), resveratrol (6), gallic acid (9) and its ethylester (5), β-sitosterol glucoside (10) and 13b-hydroxy-pheophorbide a (8). This study gives some concrete evidence to support the ethnopharmacological use of Mezoneuron benthamianum leaves extract in the management of malaria. The active compounds can be further studied for their antiplasmodial potential, as well as their suitability to be used as quality markers for the standardization of this herbal drug from the Guinean traditional pharmacopeia.


Negative feelings and the desire to eat in bulimia nervosa.
This study examines a broad range of negative feelings as possible antecedents of binge eating in bulimia nervosa (BN). Another goal is to explore the connection between negative feelings and the desire to eat as recorded continuously during two consecutive days. This is the first study comparing data from BN patients with a relevant clinical control group. Forty female BN patients, 40 female panic disorder (PD) patients, and 40 healthy women continuously recorded their feelings and the desire to eat while in their natural environment. Both patient groups reported more negative feelings than the healthy controls. BN patients had higher within-subject correlations between most negative feelings and the desire to eat than the two control groups. BN patients rated most feelings more negatively in the hour prior to binge eating than during the rest of the day. BN patients' general mood state worsened after binge eating but returned to prebinge levels after purging. The study provides additional evidence that unspecific negative feelings play an important role in the context of binge-eating behavior in BN.


LaVO4: Eu3+ nano-islands onto silica for achieving fluorescence enhancement and their detection of Fe3+ ions and anti-counterfeiting applications.
Rare earth (RE) composite fluorescent materials are favored by researchers in the field of anti-counterfeiting and ion sensing due to their fascinating optical properties. Ultra-small RE fluorescent nanoparticles are anchored on inorganic carriers by a simple preparation method to improve luminous intensity and hydrophilicity, which has not been explored yet. Herein, LaVO4: Eu3+ nano-islands anchored on silica with high fluorescence intensity and easy formation of stable colloidal solution is designed. Through a simple and mild hydrothermal approach, ultra-small LaVO4: Eu3+ nano-islands are highly dispersed on the surface of hierarchical hollow silica sphere (HHSS) to expose more luminescent centers. Remarkably, the stable HHSS@LaVO4: Eu3+ colloidal solution displayed highly sensitive and selective sensor for Fe3+ ions. The "island-sea synergy" structure formed by the LaVO4: Eu3+ nano-islands and the surrounding silica surface makes HHSS@LaVO4: Eu3+ to be an outstanding sensor for the effective detection of iron ions in water. In addition, HHSS@LaVO4: Eu3+ phosphor exhibit unique properties for anti-counterfeiting and encryption applications. These findings provide a promising strategy for the carrierisation of RE luminescent materials to improve optical properties and enable broader applications.


Pathogenesis of hemophagocytic syndrome (HPS).
Hemophagocytic syndrome (HPS) is a clinicopathologic entity characterized by increased proliferation and activation of benign macrophages with hemophagocytosis throughout the reticuloendothelial system. Uncontrolled T-lymphocyte activation is responsible for increased T(H)1 cytokines secretion such as IFN-gamma, IL-12 and IL-18 that promotes macrophage activation. Genetic defects specific for cytotoxic T lymphocytes (CTL) and natural killer (NK) cells have been identified in patients with primary HPS that are responsible for altered cell death and apoptosis induction or target killing. HPS may be secondary to malignancy, infection or autoimmune disease, and mechanisms involved are poorly understood. However, in adult-onset Still's disease, juvenile chronic arthritis and probably systemic lupus erythematosus, IL-18 might play a role in initiating macrophage activation.


A new prostaglandin E1 analogue (TFC-612) prevents a decrease in motor nerve conduction velocity in streptozocin-diabetic rats.
A new prostaglandin E1 analogue (TFC-612) was orally given to streptozocin-diabetic rats for 4 weeks after the induction of diabetes and its effects on motor nerve conduction velocity were studied. The compound significantly prevented a decrease of the velocity but did not reverse abnormal sorbitol and myo-inositol contents of the sciatic nerve. The results suggest that TFC-612 has a potent effect on diabetic nerve dysfunction via other mechanism than the correction of sorbitol and myo-inositol metabolisms and could be a potential compound for therapy of diabetic polyneuropathy.


Antibodies to histones in infectious mononucleosis.
A polyspecific human monoclonal (auto)antibody, isolated from a patient in the acute phase of infectious mononucleosis, was found to react with all subfractions (H1, H2A, H2B, H3 and H4) of histones. This finding prompted us to study the occurrence of antibodies to histones in sera of patients with infectious mononucleosis. It was found that IgM binding to histones was detectable both in control and patient sera; however, sera from patients showed binding values of IgM antibodies to histones significantly higher than those of healthy controls; moreover, both in control and patient groups anti-histone IgM activity was found to correlate with serum IgM concentration. These findings suggest that anti-histone IgM antibodies belong to the class of antibodies defined as "natural antibodies" and that their increase during infectious mononucleosis is due to Epstein-Barr virus-induced polyclonal B cell activation.


"No one told me anything about it and I cannot explain it": Illness perception in symptomatic and asymptomatic patients with cancer-associated thrombosis.
Patients with cancer, have reported cancer-associated thrombosis (CAT), a distressing event in their overall illness. However, whether the clinical presentation of CAT; symptomatic versus asymptomatic, impacts illness perception is poorly elucidated. The aim of this study was to explore illness perception in patients with CAT, stratified by the clinical presentation. In a qualitative design, we conducted a three-step workshop. Patients were included from a specialised cardiology care unit for oncology patients. Data analysis was performed using framework analysis. The analytic framework was based on the five components of illness perception: (1) identity of illness, (2) causal beliefs, (3) timeline beliefs, (4) beliefs about control/cure and (5) consequences. Elleven patients with CAT participated in the workshop; five symptomatic and six asymptomatic. Whitin each category of illness perception following notions emerged (1) the identity of CAT was only tangible for symptomatic participants, (2) the aetiology was considered important information for symptomatic participants, which was in contrast to asymptomatic participants, (3) asymptomatic participant did not consider recurrent CAT a threat towards their health, (4) asymptomatic participants were prone to information overload, whilst information was imperative to the sense of control in symptomatic participants, (5) low molecular weight heparin treatment was accepted in symptomatic participants due to remission of symptoms. The clinical presentation of CAT (asymptomatic/symptomatic) proved essential to illness perception. These findings indicate that information level and communication within the medical consultation, should actively consider the clinical presentation of CAT in order to optimize management and compliance.


[Extrachromosomal antibiotic resistance of the intestinal bacterial flora. II. The role of resistance acquired by endogenous intestinal bacterial flora (author's transl)].
In a previous paper, we have shown that multiple resistant bacteria in the human gut flora may have two origins: firstly, through food which carries resistant bacteria and secondly, our present study, resident intestinal bacteria which become resistant to antibiotics, either because they are chromosomic mutant or because they acquire resistance plasmid from an exogen strain. In all cases these resistant bacteria can implant and multiply in the gut only if there are disorders in the normal bacterial equilibrium. Antibiotherapy can create this inbalance and especially if it is not carefully followed up: it selects resistant bacteria, allows their implantation, and promotes plasmid R transfer. The resistant bacteria are able, in some cases, to spread out from the intestinal tract and give rise to further infections.


Haptoglobin is frequently low in patients with myelofibrosis: Clinical relevance.
A recent study, showing the absence of paroxysmal nocturnal hemoglobinuria clones in myelofibrosis, has reopened the debate around the role of decreased haptoglobin in this disease. We present here a large prospective analysis of the clinical significance of low haptoglobin in 152 patients with myelofibrosis. Low haptoglobin (<32mg/dL) was observed in 50 patients (33%). Decreased haptoglobin did not associate with low hemoglobin levels, positive Coombs test or abnormal liver function tests, suggesting it is not result of autoimmune hemolytic anemia or liver cirrhosis. Factors strongly correlating with decreased haptoglobin were high JAK2 allele burden and ongoing treatment with JAKi. Larger scale serial measurement and longer follow-up is needed to further explain our findings.


The influence of scaffold material on chondrocytes under inflammatory conditions.
Cartilage tissue engineering aims to repair damaged cartilage tissue in arthritic joints. As arthritic joints have significantly higher levels of pro-inflammatory cytokines (such as IL-1β and TNFα that cause cartilage destruction, it is critical to engineer stable cartilage in an inflammatory environment. Biomaterial scaffolds constitute an important component of the microenvironment for chondrocytes in engineered cartilage. However, it remains unclear how the scaffold material influences the response of chondrocytes seeded in these scaffolds under inflammatory stimuli. Here we have compared the responses of articular chondrocytes seeded within three different polymeric scaffolding materials (silk, collagen and polylactic acid (PLA)) to IL-1β and TNFα. These scaffolds have different physical characteristics and yielded significant differences in the expression of genes associated with cartilage matrix production and degradation, cell adhesion and cell death. The silk and collagen scaffolds released pro-inflammatory cytokines faster and had higher uptake water abilities than PLA scaffolds. Correspondingly, chondrocytes cultured in silk and collagen scaffolds maintained higher levels of cartilage matrix than those in PLA, suggesting that these biophysical properties of scaffolds may regulate gene expression and the response to inflammatory stimuli in chondrocytes. Based on this study we conclude that selecting the proper scaffold material will aid in the engineering of more stable cartilage tissues for cartilage repair, and that silk and collagen are better scaffolds in terms of supporting the stability of three-dimensional cartilage under inflammatory conditions.


Streptococcus suis-Associated Meningitis, Bali, Indonesia, 2014-2017.
Streptococcus suis is an emerging agent of zoonotic bacterial meningitis in Asia. We describe the epidemiology of S. suis cases and clinical signs and microbiological findings in persons with meningitis in Bali, Indonesia, using patient data and bacterial cultures of cerebrospinal fluid collected during 2014-2017. We conducted microbiological assays using the fully automatic VITEK 2 COMPACT system. We amplified and sequenced gene fragments of glutamate dehydrogenase and recombination/repair protein and conducted PCR serotyping to confirm some serotypes. Of 71 cases, 44 were confirmed as S. suis; 29 isolates were serotype 2. The average patient age was 48.1 years, and 89% of patients were male. Seventy-seven percent of patients with confirmed cases recovered without complications; 11% recovered with septic shock, 7% with deafness, and 2% with deafness and arthritis. The case-fatality rate was 11%. Awareness of S. suis infection risk must be increased in health promotion activities in Bali.


Anti-inflammatory activity of phylligenin, a lignan from the fruits of Forsythia koreana, and its cellular mechanism of action.
The fruits of Forsythia koreana have long been used in Chinese medicine to treat inflammatory disorders. However, the pharmacological data is not sufficient to clearly establish a scientific rationale for the anti-inflammatory medicinal use of this plant material, and the search for its active principles has been limited so far. Phylligenin (lignan) was isolated from the fruits of Forsythia koreana and its anti-inflammatory activity was examined. Phylligenin (1-100 microM) and the methanol extract of Forsythia koreana fruits inhibited cyclooxygenase-2 (COX-2)-mediated prostaglandin E(2) and inducible nitric oxide synthase (iNOS)-mediated nitric oxide (NO) synthesis from lipopolysaccharide-treated RAW 264.7 cells. In the mechanism study, phylligenin inhibited iNOS expression and nuclear factor-kappaB (NF-kappaB) activation but had no effect on COX-2 expression. Moreover, phylligenin significantly inhibited mouse carrageenan-induced paw edema by intraperitoneal administration (22.1-34.7% inhibition at 12.5-100 mg/kg). These pharmacological properties indicate that phylligenin possesses significant anti-inflammatory activity in vitro and in vivo, and may provide the scientific rationale for anti-inflammatory use of the fruits of Forsythia koreana.


Mobilization of mitochondrial Ca2+ by hydroperoxy-eicosatetraenoic acid.
Hydroperoxy-eicosatetraenoic acids (HPETEs) and less effectively, also hydroxy-eicosatetraenoic acids (HETEs) stimulated Ca2+ release from rat liver mitochondria. Ca2+ release is accompanied by intramitochondrial pyridine nucleotide oxidation and hydrolysis. Both Ca2+ release and pyridine nucleotide oxidation are impeded when the flow of electrons between pyridine nucleotides and HPETE is impaired. Measurements of the mitochondrial membrane potential indicate that HPETE-stimulated Ca2+ release is not due to uncoupling of mitochondria. It is suggested that HPETEs and HETEs may act as mobilizers of mitochondrial Ca2+ during signal transduction related to proliferation and tumor promotion.


Methods for assessing splenic macrophage depletion by liposome encapsulated clodronate.
Small unilamellar vesicles containing clodronate (SUVc) injected intravenously will deplete splenic macrophages and the degree of histological depletion can be assessed by determining the clearance and uptake of monoclonal antibody coated erythrocytes. Splenic Fc dependent clearance, assessed in decomplemented animals, provides a more sensitive index of the effects of large multilamellar liposome encapsulated clodronate (MLVc) and SUVc than does the clearance of complement coated erythrocytes on macrophage depletion in the spleen. MLVc were more efficient than SUVc in inducing a reduction in the number of red pulp macrophages within the spleen. Receptor specific red cell uptake in the spleen could be used as an alternative to histology when assessing splenic macrophage depletion. Encapsulation of clodronate is crucial to its depleting effect since the free drug in saline does not change splenic macrophage number or function.


Positron Emission Tomography for pretreatment staging and posttreatment evaluation in cancer of the anal canal.
In recent years, combined modality therapy (CMT) with chemotherapy and radiation has replaced surgery as the preferred treatment for cancer of the anal canal. Clinical staging with computed tomography (CT) scan alone may underestimate the extent of disease. We investigated the utility of positron emission tomography (PET) with 2-deoxy-2[F-18]fluoro-D-glucose (FDG) in the staging and determination of response to CMT. From September 1999 to August 2002, 21 patients with cancer of the anal canal were studied prospectively. All patients underwent pretreatment PET, in addition to standard clinical evaluation that included CT scanning. Follow-up PET studies were ordered one month after completion of CMT. Sites of metastases not observed on CT scan were identified in five of 21 patients (24%). These sites included pelvic lymph nodes (four patients) and distant omental metastasis (one patient). In another patient, PET confirmed the presence of suspected M(1) disease in the liver. Posttreatment PET imaging was less useful. Nine patients had minimal residual PET activity at the primary site on the one-month follow-up PET study, but only three of these subsequently developed local recurrence. In addition, recurrences occurred in three patients (two local, one distant) of the six who had negative posttreatment PET studies. FDG-PET, in conjunction with CT scanning, provides additional staging information in cancer of the anal canal. This information may have implications for prognosis and radiotherapy planning. Posttreatment PET scans appear to be of little value in predicting durability of response.


Endogenous myelin basic protein-serum factors (MBP-SFS) and anti-MBP antibodies in a patient with post-herpes simplex virus acute disseminated encephalomyelitis.
The measurement of myelin basic protein serum factors (MBP-SFs) and anti-MBP antibodies in specimens from a patient with post-herpes simplex acute disseminated encephalomyelitis (ADE) is described. Transitory appearance of high affinity anti-MBP antibodies in the absence of detectable MBP-SFs was observed. This pattern was similar to that found previously in acute multiple sclerosis and experimental allergic encephalomyelitis. These findings are consistent with the hypothesis that a possible normal role of MBP-SFs is as neuroautotolerogens, preventing autoreactive lymphocyte clones from damaging myelin.


Potentiated nephrotoxicity of cisplatin when combined with amikacin comparing young and adult rats.
We compared the nephrotoxic interaction between cisplatin (CP) and amikacin (AM) in young and adult rats, using different dosage combinations. Following a single i.v. dose of CP, AM was administered s.c. for 14 days. The dose of CP was chosen to cause a 20%-50% fall in the glomerular filtration rate (GFR), while a dose of AM was chosen that had only a minimal effect on GFR. In adult rats, a decrease in GFR to 60% of the control value after CP alone was seriously aggravated by a non-toxic AM course given during 2 weeks after CP. In this combination, the GFR per 100 g body weight was reduced to 30% of control at week 2, which rose to 40% of control at week 15. In young rats, a non-toxic AM course did not aggravate the CP-induced impairment in GFR. However, when the dose of AM was increased to cause a 20% reduction in the GFR, the nephrotoxicity was potentiated. When measured at week 2, the GFR per 100 g body weight was 40% of control after the combined treatment compared with 80% of control after CP alone. As in adult rats, there was only a partial recovery of the GFR. In conclusion, in both adult and young rats, a course of AM following a single injection of CP potentiated CP-induced nephrotoxicity.


Demographic models of the northern spotted owl (Strix occidentalis caurina).
Calassical demographic methods applied to life history data on the northern spotted owl yield and estimate of the annual geometric rate of increase for the population of λ=0.96±0.03, which is not significantly different from that for a stable population (λ=1.00). Sensitivity analysis indicates that adult annual survivorship has by far the largest influence on λ, followed by the probability that juveniles survive dispersal, and the adult annual fecundity. Substantial temporal fluctuations in demographic parameters have little effect on the long-run growth rate of the population because of the long adult life expectancy. A model of dispersal and territory occupancy that assumes demographic equilibrium is evaluated using data on the amount of old forest habitat remaining in the Pacific Northwest and the current occupancy of this habitat by northern spotted owls. This model is employed to predict the effect of future habitat loss and fragmentation on the population, implying that extinction will result if the old forest is reduced to less than a proportion 0.21±0.02 of the total area in a large region. The estimated minimum habitat requirement for the population is greater than that allowed in management plants by the USDA Forest Service.


TIS gene expression in cultured rat astrocytes: multiple pathways of induction by mitogens.
Accumulation of TIS1 and TIS11 (Lim et al.: Oncogene 1:263-270, 1987) mRNAs in secondary cultures of rat neocortical astrocytes was much greater in response to tetradecanoyl phorbol acetate (TPA) than in response to either epidermal growth factor (EGF) or fibroblast growth factor (FGF). In contrast, EGF, FGF, and TPA were equally effective in inducing accumulation of TIS8 and TIS28/c-fos mRNAs. These data suggested that TPA and the polypeptide mitogens might induce TIS gene expression by distinct pathways. When maximally inducing concentrations of EGF and FGF were co-administered to astrocyte cultures, TIS mRNA accumulations were no greater than those observed for the individual growth factors, suggesting that EGF and FGF saturate a common, limiting step in their induction pathways. In contrast, when either EGF or FGF was presented to astrocytes in combination with maximally inducing levels of TPA, the resulting levels of accumulation of TIS mRNAs were at least as great as the sum of the levels induced by the individual mitogens. Stimulation of [3H]-thymidine incorporation demonstrated an identical pattern of interaction; EGF and FGF co-administration was no more effective than either polypeptide mitogen alone, but, when presented to astrocyte cultures along with maximally inducing concentrations of TPA, either EGF or FGF was able to increase incorporation of [3H]-thymidine. Superinduction of all the TIS genes occurred if cycloheximide (CHX) was present during TPA exposure. Once again, two distinct classes of responses of the various TIS genes occurred; superinduction of TIS1, TIS7, TIS11, and TIS28/c-fos mRNA accumulation ranged from 10- to 20-fold, while CHX superinduction of TIS8 and TIS10 was far more modest, ranging from 2- to 3-fold.(ABSTRACT TRUNCATED AT 250 WORDS)


Properties of residual catalase in the erythrocytes of Japanese-type acatalasemia.
In Japanese-type acatalasemia erythrocytes, the presence and properties of residual catalase were determined and compared with those of normal erythrocyte catalase. Residual catalase activity was proved by titration, active staining after polyacrylamide gel electrophoresis, and measurement of oxygen evolution. Residual catalase protein, demonstrated by double immunodiffusion, was similar to that of normal catalase. The properties of residual catalase activity were identical with those of normal catalase activity. It occurred as three fractions of equal specific activity by DEAE column chromatography. These observations suggest that Japanese-type acatalasemia contains residual catalase with properties similar to those of normal catalase.


Occurrence and mass balance of emerging brominated flame retardants in a municipal wastewater treatment plant.
As a class of alternative flame retardants, "novel" brominated flame retardants (NBFRs) have been widely used in clothing, textiles, electronics, building materials, and plastics and are frequently found in environmental samples. Although the use and discharge of NBFRs are increasing all over the world, little information is available about their fates and removal in wastewater treatment plants (WWTPs). This study investigated the occurrence and behavior of 2,4,6-tribromophenyl allyl ether (ATE), 2,3-dibromopropyl tribromophenyl ether (DPTE), tetrabromo-o-chlorotoluene (TBCT), pentabromobenzyl acrylate (PBBA), 1,2-bis(2,4,6-tribromophenoxy) ethane (BTBPE), bis(2-ethylhexyl)-3,4,5,6-tetrabromo-phthalate (TBPH), and decabromodiphenyl ethane (DBDPE) in a municipal WWTP in Beijing, China. Four of the NBFRs (ATE, BTBPE, TBPH, and DBDPE) were detected in all wastewater and biosolid samples. The dominant compound in both wastewater and biosolid samples was DBDPE, with concentrations in the ranges of 6.4-18 ng/L and 83-288 ng/g dry weight (dw), respectively. A mass flow analysis indicated that the overall removal efficiencies were 31±18% for ATE, 97±1.5% for DPTE, 79±11% for BTBPE, 87±5.3% for TBPH, and 93±3.3% for DBDPE, but the removal efficiency in aqueous phase only ranged from -43 (ATE) to 57% (DBDPE). The low and negative removal efficiencies of ATE were possibly due to the transformation of DPTE during the anaerobic treatment processes. Under the aerobic conditions, BTBPE, TBPH and DBDPE underwent a very slow biodegradation. A total of 68-91% of the initial mass loadings of ATE, BTBPE, TBPH and DBDPE were found in the biosolid samples, suggesting that the four NBFRs were removed mainly via sorption in the WWTP.The biosolid was a huge reservoir of target NBFRs, and effective removal of NBFRs during treatment in the WWTP is critical.


A comparison of Covid-19 cases and deaths in Turkey and in other countries.
In this study, the characteristics of the Covid-19 pandemic in Turkey are examined in terms of the number of cases and deaths, and a characteristic prediction is made with an approach that employs artificial intelligence. The number of cases and deaths are estimated using the number of tests, the numbers of seriously ill and recovered patients as parameters. The machine learning methods used are linear regression, polynomial regression, support vector regression with different kernel functions, decision tree and artificial neural networks. The obtained results are compared by calculating the coefficient of determination (R 2), and the mean absolute percentage error (MAPE) values. When R 2 and MAPE values are compared, it is seen that the optimal results for cases in Turkey are obtained with the decision tree, for deaths with polynomial regression method. The results reached for the United States of America and Russia are similar and the optimal results are obtained by polynomial regression. However, while the optimal results are obtained by neural networks in the Indian data, linear regression for the cases in the Brazilian data, neural network for the deaths, decision tree for the cases in France, polynomial regression for the deaths, neural network for the cases in the UK data and decision tree for the deaths are the methods that produced the optimal results. These results also give an idea about the similarities and differences of country characteristics.


In vivo cytogenetic effects of a commercially formulated mixture of cypermethrin and quinalphos in mice.
In vivo cytogenetic effects of commercially formulated cypermethrin (CYP, synthetic pyrethroid insecticide) and/or quinalphos (QUI, organophosphate insecticide), generally used in combination, were examined through chromosomal aberrations (CA) and micronucleus test (MT) in mice. Male mice were orally gavaged to a single dose of CYP/QUI commercial mixture (22, 44 or 67 mg/kg b.wt.) for 24h (CA) or 48 h (MT). Based on the concentrations of active ingredients of CYP and QUI present in the test doses of CYP/QUI mixture, mice were orally exposed to 0.66, 1.32 and 2 mg/kg of CYP or 4.4, 8.8 and 13.4 mg/kg of QUI. For reference, a group of five mice was intraperitoneally administered to cyclophosphamide (20 or 50 mg/kg) or orally gavaged to peanut oil for vehicle control. Exposure of CYP/QUI mixture inhibited the mitotic index (MI) and induced CA in a dose-dependent manner at 24 h; however, significant (p<0.01 or 0.001) frequencies of CA were observed at 44 mg/kg onwards, whereas inhibition of MI at 67 mg/kg. Independent exposure of QUI at 8.8 mg/kg onwards also significantly (p<0.01 or 0.001) inhibited MI and induced CA, whereas CYP at 2 mg/kg (highest concentration in CYP/QUI mixture) inhibited MI significantly but failed to induce CA. Chromatid breaks and fragments found to be frequent aberrations in all the test groups. Treatment of CYP/QUI mixture also induced micronucleus formation dose-dependently at 48 h, yet statistically significant (p<0.001) frequencies of micronucleated polychromatic erythrocytes (MNPCE) were observed at 44 mg/kg onwards. QUI (8.8 and 13.4 mg/kg) alone also induced significant frequencies of MNPCE, whereas frequencies of MNPCE observed with the CYP even at 2 mg/kg were comparable to that of vehicle control. Present findings indicate the genotoxicity potential of CYP/QUI mixture and suggest that the simultaneous presence of the toxic doses of CYP and QUI can lead to synergistic genotoxicity in mice and may pose mutagenic risk in human beings.


Outcomes of radical surgical resections for nonmetastatic anorectal melanomas: Experience from a tertiary care centre.
The outcome of radical surgery in nonmetastatic anorectal melanoma (AM) patients is studied infrequently. Here, we aimed to explore the stage-wise outcomes and the impact of radical resections in these patients. In this single-centre retrospective study, data of 154 eligible patients were recorded and analysed. Data were obtained from November 2010 to September 2019 with follow-up until November 2020. Overall survival (OS) and disease-free survival (DFS) was calculated by Kaplan Meir method and univariate analysis of prognostic factors by Cox regression. Of 154 patients, 110 were metastatic (stage III) and 44 were nonmetastatic (stage I:22, stage II:22) and underwent curative resections. Median follow-up was 48 months (14-119 months). A total of 39 patients underwent total mesorectal excisions (TME) and five transanal excision (TAE) were performed. Seven patients underwent extended resections. Stage I and II patients had 3- and 5-year OS of 40% and 36%; and DFS of 45% and 33.2%, respectively. Median OS and DFS were 31 and 24 months, respectively. Stage II (node-positive) patients had better median OS compared to stage III (21 vs. 4 months; p = 0.000), and 54.5% patients had recurrences, most commonly both systemic and nodal (45.83%). Median OS of patients without recurrence was 34 months. In this large surgical series of AMs, outcome in stage I and II patients was significantly better than stage III and patients with stage II disease can have acceptable oncological outcomes. Radical surgical resections with or without lymphadenectomy could be considered in these patients. The role of adjuvant systemic therapy and radiation needs to be explored as part of multimodality treatment.


[Thrombotic microangiopathy : Relevant new aspects for intensive care physicians].
Thrombotic microangiopathy (TMA) is a clinical syndrome that is characterized by hemolysis, thrombocytopenia, and acute kidney injury, known as atypical hemolytic syndrome (aHUS), thrombotic thrombocytopenic purpura (TTP), and shigatoxin-associated HUS (STEC-HUS) among others. Several diseases, like malignoma, infections, malignant hypertension, or autoimmune disease can result in secondary TMAs. aHUS is caused by a hyperactivated complement system. Identification of the underlying causes of the TMA is the most important issue and directly associated with treatment success. In case of secondary TMAs, treatment of the actual disease is the most important step, while in case of complement-mediated HUS treatment of choice is plasma exchange or anticomplement agents. For the treatment of TTP, rapid initiation of plasma exchange or plasma infusion is the treatment of choice. Patients with STEC-HUS should solely receive supportive treatment.


Effects of drinking water temperature in winter on growth performance, water consumption, surface temperature, and intestinal development of geese from 21 to 49 days of age.
This research aimed to investigate the effects of drinking water temperatures on growth performance, water consumption, surface temperature, organ indices, blood parameters, and intestinal development of geese, and determine the optimal drinking water temperature for 21 to 49-d geese. A total of 192 twenty-one-day male Yuzhou white geese were allocated randomly to 4 groups with 8 replicate pens per group according to the drinking water temperature (drinking water temperature [7℃-12℃] at ambient temperature [TC], 18℃ [T1], 27℃ [T2], and 36℃ [T3], respectively). The results showed that increasing drinking water temperature did not significantly improve the BW, ADG, and ADFI of geese (P > 0.05), whereas drinking warm water of 36℃ for geese had a trend to increase FCR (0.05 < P < 0.1). Geese in group T2 drank the most water per day on average, whereas geese in group TC drank the least (P < 0.001). Geese in groups T1, T2, and T3 had higher eyes temperatures than group TC (P < 0.001). No significant differences were found on the organ indices and blood biochemical parameters (P > 0.05). Geese from group T1 had higher crypt depth and muscularis thickness of duodenum (P < 0.05), and lower ratio of villus height to crypt depth than other groups (P < 0.001). Geese from group T1 had higher activities of trypsin in duodenum and jejuna and amylase in jejuna at 49 d than other groups (P < 0.01). Overall, these data indicate drinking water at 18℃ can increase water consumption and eyes temperature, and improve the activity of digestive enzymes and promote intestinal development. Under our experimental conditions, we recommend that the optimal drinking water temperature for geese from 21 to 49 d of age is 18°C.


Identification of possible sources for potentially toxic elements and polycyclic aromatic hydrocarbons and their spatially varying relationships in urban soils of Dublin, Ireland.
Potentially toxic elements (PTEs) and polycyclic aromatic hydrocarbons (PAHs) harm the ecosystem and human health, especially in urban areas. Identifying and understanding their potential sources and underlying interactions in urban soils are critical for informed management and risk assessment. This study investigated the potential sources and the spatially varying relationships between 9 PTEs and PAHs in the topsoil of Dublin by combining positive matrix factorisation (PMF) and geographically weighted regression (GWR). The PMF model allocated four possible sources based on species concentrations and uncertainties. The factor profiles indicated the associations with high-temperature combustion (PAHs), natural lithologic factors (As, Cd, Co, Cr, Ni), mineralisation and mining (Zn), as well as anthropogenic inputs (Cu, Hg, Pb), respectively. In addition, selected representative elements Cr, Zn, and Pb showed distinct spatial interactions with PAHs in the GWR model. Negative relationships between PAHs and Cr were observed in all samples, suggesting the control of Cr concentrations by natural factors. Negative relationships between PAHs and Zn in the eastern and north-eastern regions were related to mineralisation and anthropogenic Zn-Pb mining. In contrast, the surrounding regions exhibited a natural relationship between these two variables with positive coefficients. Increasing positive coefficients from west to east were observed between PAHs and Pb in the study area. This special pattern was consistent with prevailing south-westerly wind direction in Dublin, highlighting the predominant influences on PAHs and Pb concentrations from vehicle and coal combustion through atmospheric deposition. Our results provided a better understanding of geochemical features for PTEs and PAHs in the topsoil of Dublin, demonstrating the efficiency of combined approaches of receptor models and spatial analysis in environmental studies.


Predictive items of 2-year mortality in nonagenarians with cognitive impairment at baseline: the NonaSantfeliu study.
We aimed to identify a set of predictors of mortality at 2-year follow-up in a cohort of nonagenarians with cognitive impairment at baseline. The following variables were evaluated; functional status determined by the indices of Barthel Index for basic and Lawton and Brody for instrumental activities of daily living, while cognition was assessed using the Spanish version of the mini-mental state examination (MEC). Cognitive impairment was graded as severe for those having MEC scores of 0-17 and mild for scores of 18-23. A total of 77 nonagenarians with a MEC score of 23 or below were included: 61 women (79.25%) and 16 men, with a mean age of 93.7 (3.4) years. The rate of mortality was 55.8%. According to MEC scores the proportion of deaths during the 2-year follow-up period ranged from 35.2% (6 out of 17) among mildly impaired participants to 61.6% (37 out of 60) among severely impaired participants (p=0.05). Multivariate regression analysis showed that the risk of dying in this cohort was associated with being older (p<0.01, OR 1.23, 95% CI 1.03-1.4) and having a previous history of heart failure (p<0.0001, OR 8.72, 95% CI 2.66-28.5). In conclusion age and heart failure predict mortality among nonagenarians with prior cognitive impairment.


Erratum: N6-methyladenosine-induced ERRγ triggers chemoresistance of cancer cells through upregulation of ABCB1 and metabolic reprogramming: Erratum.
[This corrects the article DOI: 10.7150/thno.40144.].


Disposition of guanadrel in subjects with normal and impaired renal function.
The disposition of a single 25 mg oral dose of guanadrel was evaluated in 22 subjects with various degrees of renal function. The terminal elimination half-life was significantly prolonged in subjects with a creatinine clearance (ClCr) less than 30 mL/min/1.73 m2 (19.2 +/- 16.8 h) compared to 3.7 +/- 1.9 h in subjects with a ClCr greater than 80 mL/min/1.73 m2. Apparent total body clearance (Clp/F) was also progressively lower in the patients with decreased renal function and the decline was significantly correlated with ClCr (Clp/F = 0.0294 + 0.0236 Clcr, r = 0.74, P = 0.002). Renal clearance and apparent nonrenal clearance also declined as creatinine clearance decreased, and both were significantly correlated with the observed ClCr. Apparent volume of distribution averaged 11.5 +/- 8.9 L/kg and did not differ in patients with decreased renal function compared to those with normal renal function. Thus, the disposition of guanadrel is significantly altered in the presence of renal insufficiency and dosage adjustments may be necessary, especially in patients with ClCr less than 50 ml/min.


Expression and function of inwardly rectifying potassium channels after experimental subarachnoid hemorrhage.
Cerebral vasospasm after subarachnoid hemorrhage (SAH) is because of smooth muscle contraction, although the mechanism of this contraction remains unresolved. Membrane potential controls the contractile state of arterial myocytes by gating voltage-sensitive calcium channels and is in turn primarily controlled by K(+) ion conductance through several classes of K(+) channels. We characterized the role of inwardly rectifying K(+) (K(IR)) channels in vasospasm. Vasospasm was created in dogs using the double-hemorrhage model of SAH. Electrophysiological, real-time quantitative reverse-transcriptase polymerase chain reaction, Western blotting, immunohistochemistry, and isometric tension techniques were used to characterize the expression and function of K(IR) channels in normal and vasospastic basilar artery 7 days after SAH. Subarachnoid hemorrhage resulted in severe vasospasm of the basilar artery (mean of 61% +/- 5% reduction in diameter). Membrane potential of pressurized vasospastic basilar arteries was significantly depolarized compared with control arteries (-46 +/- 1.4 mV versus -29.8 +/- 1.8 mV, respectively, P < 0.01). In whole-cell patch clamp of enzymatically isolated basilar artery myocytes, average K(IR) conductance was 1.6 +/- 0.5 pS/pF in control cells and 9.2 +/- 2.2 pS/pF in SAH cells (P = 0.007). Blocking K(IR) channels with BaCl(2) (0.1 mmol/L) resulted in significantly greater membrane depolarization in vasospastic compared with normal myocytes. Expression of K(IR) 2.1 messenger ribonucleic acid (mRNA) was increased after SAH. Western blotting and immunohistochemistry also showed increased expression of K(IR) protein in vasospastic smooth muscle. Blockage of K(IR) channels in arteries under isometric tension produced a greater contraction in SAH than in control arteries. These results document increased expression of K(IR) 2.1 mRNA and protein during vasospasm after experimental SAH and suggest that this increase is a functionally significant adaptive response acting to reduce vasospasm.


Control of replication of FII plasmids: comparison of the basic replicons and of the copB systems of plasmids R100 and R1.
The copy numbers of the FII plasmids R1 and R100 were determined in four different ways and found to be identical. Deletion of one of the copy number control genes, copB, together with its promoter gives rise to plasmid copy mutants with an increased copy number. The increase was found to be 8- and 3.5-fold for plasmids R1 and R100, respectively. These deletion derivatives were found to be extremely sensitive to the presence of CopB activity from their own parent plasmid but not to that of the other plasmid. Hence, the CopB protein and its target are plasmid-specific and not FII-group-specific. These results are consistent with the high degree of nonhomology between plasmids R1 and R100 in a 250-bp region covering the distal part of the copB gene and the repA promoter region, which contains the target for the CopB protein.


Repeated positioning in accurate radiotherapy based on virtual net technique and contrary reconstruction scheme.
This paper presents a new repeated positioning system for external radiotherapy. A new scheme is proposed to rebuild patient's body surface 3-D image based on simple stereo vision model and virtual net technique, which improves the reconstruction precision of the template. For the calculation of the positioning error, the contrary reconstruction scheme is adopted and the positioning speed is greatly improved. The 3-D reconstructed video image of the right position in the first positioning is used as the reference template for the next positioning, and the positioning error is evaluated by projecting the template image into the patient's real-time video images and calculating the correlation ratio in the areas limited by the triangle of the reference image.


Arthroscopic surgery of the temporomandibular joint: report of 64 patients with long-term follow-up.
One hundred temporomandibular joints (64 patients) were examined arthroscopically. Complications from the procedure were low (4%). Eighty joints (50 patients) received superior compartment sweep and lavage. Patients with anteriorly displaced nonreducing discs (closed lock) had the highest success rate (83%) from this procedure when evaluated 9 to 30 months later.


Regulation of epithelium-specific Ets-like factors ESE-1 and ESE-3 in airway epithelial cells: potential roles in airway inflammation.
Airway inflammation is the hallmark of many respiratory disorders, such as asthma and cystic fibrosis. Changes in airway gene expression triggered by inflammation play a key role in the pathogenesis of these diseases. Genetic linkage studies suggest that ESE-2 and ESE-3, which encode epithelium-specific Ets-domain-containing transcription factors, are candidate asthma susceptibility genes. We report here that the expression of another member of the Ets family transcription factors ESE-1, as well as ESE-3, is upregulated by the inflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in bronchial epithelial cell lines. Treatment of these cells with IL-1beta and TNF-alpha resulted in a dramatic increase in mRNA expression for both ESE-1 and ESE-3. We demonstrate that the induced expression is mediated by activation of the transcription factor NF-kappaB. We have characterized the ESE-1 and ESE-3 promoters and have identified the NF-kappaB binding sequences that are required for the cytokine-induced expression. In addition, we also demonstrate that ESE-1 upregulates ESE-3 expression and downregulates its own induction by cytokines. Finally, we have shown that in Elf3 (homologous to human ESE-1) knockout mice, the expression of the inflammatory cytokine interleukin-6 (IL-6) is downregulated. Our findings suggest that ESE-1 and ESE-3 play an important role in airway inflammation.


Isolation and immunochemical characterization of the major allergen of birch pollen (Betula verrucosa).
The classification of some of the extractable birch pollen antigens as allergens was established by crossed radioimmunoelectrophoresis (CRIE). In CRIE the major allergen (antigen 23) exhibited the strongest "radiostaining," and only a few other components of birch pollen extract were visibly radiostained. The major allergen and a preparation containing mainly the minor allergens, antigens 25 and 19, were isolated from a crude aqueous birch pollen extract by a combination of anion-exchange, size-exclusion, and chelate chromatography. Antigen 23 was purified to near homogeneity. The molecular weights and the pIs of antigens 23, 25, and 19 were determined to be 17,000 daltons, pI 5.25 (5.5, 5.0); 25,000 daltons, pI 5.0 (4.9, 5.4); and 29,000 daltons, pI 6.2 (5.4), respectively. The classification of antigen 23 as the major allergen in birch pollen was supported by results of RAST inhibition experiments, RAST screening, and skin prick testing.


A novel, high-throughput technique for species identification reveals a new species of tsetse-transmitted trypanosome related to the Trypanosoma brucei subgenus, Trypanozoon.
We describe a novel method of species identification, fluorescent fragment length barcoding, based on length variation in regions of the 18S and 28Salpha ribosomal DNA. Fluorescently tagged primers, designed in conserved regions of the 18S and 28Salpha ribosomal DNA, were used to amplify fragments with inter-species size variation, and sizes determined accurately using an automated DNA sequencer. By using multiple regions and different fluorochromes, a barcode unique to each species was generated. The technique was developed for the identification of African tsetse-transmitted trypanosomes and validated using DNA from laboratory isolates representing known species, subspecies and subgroups. To test the methodology, we examined 91 trypanosome samples from infected tsetse fly midguts from Tanzania, most of which had already been identified by species-specific and generic PCR tests. Identifications were mainly in agreement, but the presence of an unknown trypanosome in several samples was revealed by its unique barcode. Phylogenetic analyses based on 18S rDNA and glycosomal glyceraldehyde phosphate dehydrogenase gene sequences confirmed that this trypanosome is a new species and it is within the Trypanosoma brucei clade, as a sister group of subgenus Trypanozoon. The overall identification rate of trypanosome-infected midgut samples increased from 78 to 96% using FFLB instead of currently available PCR tests. This was due to the high sensitivity of FFLB as well as its capacity to identify previously unrecognised species. FFLB also allowed the identification of multiple species in mixed infections. The method enabled high-throughput and accurate species identification and should be applicable to any group of organisms where there is length variation in regions of rDNA.


Pathogenic mechanism analysis of cochlear key structural lesion and phonosensitive hearing loss.
Due to ethical issues and the very fine and complex structure of the cochlea, it is difficult to directly perform experimental measurement on the human cochlea. Therefore, the finite element method has become an effective and replaceable new research means. Accurate numerical analysis on human ear using finite element method can provide better understanding of sound transmission and can be used to assess the influence of diseases on hearing and to treat hearing loss. In this research, a three-dimensional (3D) finite element model (FEM) of the human ear of cochlea was presented to investigate the destruction of basilar membrane (BM), round window (RW) sclerosis and perilymph fistula, the key structures of the cochlea, and analyze the effects of these abnormal pathological states in the cochlea on cochlear hearing, resulting in the changes in cochlear sense structure biomechanical behavior and quantitative prediction of the degree and harm of the disorder to the decline of human hearing. Therefore, this paper can deepen reader's understanding of the cochlear biomechanical mechanism and provide a theoretical foundation for clinical otology.


Vacuum cup placement during delivery--a suggested obstetric quality assessment measure.
To establish standards for the deviation of vacuum cup placement from the ideal location during operative delivery in an academic center. Data on 92 vacuum deliveries were prospectively obtained. The actual point on the newborns head was determined and both midline and anterior-posterior line deviations from the ideal point of placement were calculated. The most common indication for vacuum extraction was a nonreassuring fetal heart rate (66.7%). The average deviation on the mid anterior-posterior line was 3.72 ± 1.46 cm; the average midline-lateral deviation was 1.92 ± 1.33 cm. There was no statistically significant difference in the cup placement deviations between deliveries performed by residents and consultants. The vacuum procedure failed in 8.6% of the cases. Accurate placement of the vacuum cup on the fetal head is considered to be clinically important. This assumption requires scientific clinical proof. Our local standard for deviation was established and will serve for audit. If safer neonatal and maternal outcomes are demonstrated, the deviation from the ideal placement location ought to become a universal quality measure for vacuum deliveries.


Influence of mesoporosity on the sorption of 2,4-dichlorophenoxyacetic acid onto alumina and silica.
Two SiO2 and three Al2O3 adsorbents with varying degrees of mesoporosity (pore diameter 2-50 nm) were reacted with 2,4-dichlorophenoxyacetic acid (2,4-D) at pH 6 to investigate the effects of intraparticle mesopores on adsorption/desorption. Anionic 2,4-D did not adsorb onto either SiO2 solid, presumably because of electrostatic repulsion, but it did adsorb onto positively charged Al2O3 adsorbents, resulting in concave isotherms. The Al2O3 adsorbent of highest mesoporosity consistently adsorbed more 2,4-D per unit surface area than did the nonporous and less mesoporous Al2O3 adsorbents over a range of initial 2,4-D solution concentrations (0.025-2.5 mM) and reaction times (30 min-55 d). Differences in adsorption efficiency were observed despite equivalent surface site densities on the three Al2O3 adsorbents. Hysteresis between the adsorption/desorption isotherms was not observed, indicating that adsorption is reversible. Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy studies confirm that 2,4-D adsorption does not occur via ligand exchange, but rather via electrostatic interaction. The results indicate that adsorbent intraparticle mesopores can result in consistently greater 2,4-D adsorption, but the amount adsorbed is dependent upon surface charge and the presence of adsorbent mesoporosity. The data also suggest that when mineral pores are significantly larger than the adsorbate, they do not contribute to diffusion-limited adsorption/desorption hysteresis. Adsorbent transformations through time are discussed.


Reversible uranyl fluoride nephrotoxicity in the Long Evans rat.
Severity and duration of renal injury produced by low levels of uranyl fluoride (UO2F2) were examined in the rat. Rats received multiple ip injections of UO2F2 (cumulative dose: 0.66 or 1.32 mg U/kg body wt). Renal injury was characterized histologically by cellular and tubular necrosis of pars recta of proximal tubule (S2 and S3), with less severe cellular injury to thick ascending limb of loop of Henle and collecting tubule. Injury was evident when renal uranium levels were between 0.7 and 1.4 micrograms U/g wet kidney and was most severe when renal uranium burden was between 3.4 and 5.6 micrograms U/g. Repair of injury was rapid, with complete restoration within 35 days after exposure. Associated with injury were abnormalities in renal function, including impaired tubular reabsorption, proteinuria, and enzymuria, which appeared temporally related, to variable degrees, to progression of renal injury. Thus, reversible renal injury occurs in the rat at levels of uranium in kidney below the present Nuclear Regulatory Commission standard of 3 micrograms U/g kidney for renal injury in humans.


Temporal genetic structure of a drone congregation area of the giant Asian honeybee (Apis dorsata).
The giant Asian honeybee (Apis dorsata), like all other members of the genus Apis, has a complex mating system in which the queens and males (drones) mate at spatially defined drone congregation areas (DCAs). Here, we studied the temporal genetic structure of a DCA of A. dorsata over an 8-day time window by the genotyping of sampled drones with microsatellite markers. Analysis of the genotypic data revealed a significant genetic differentiation between 3 sampling days and indicated that the DCA was used by at least two subpopulations at all days in varying proportions. The estimation of the number of colonies which used the DCA ranged between 20 and 40 colonies per subpopulation, depending on the estimation procedure and population. The overall effective population size was estimated as high as N (e)=140. The DCA seems to counteract known tendencies of A. dorsata for inbreeding within colony aggregations by facilitating gene flow among subpopulations and increasing the effective population size.


B cells capable of spontaneous IgG secretion in cerebrospinal fluid from patients with multiple sclerosis: dependency on local IL-6 production.
Cerebrospinal fluid (CSF) from multiple sclerosis (MS) patients contains B cells capable of spontaneous IgG secretion in vitro. This study analyses the function and regulation of these cells. CSF cells obtained from nine MS patients actively produced IgG during 2-3 days in culture, and the activity decreased when CSF cells were cultured in serum-free medium. CSF cells from four controls did not secrete detectable IgG in vitro. Further experiments revealed that IL-6 played a role on MS CSF IgG-secreting cells, as can be deduced from the following findings: (i) the addition of exogenous IL-6, but not of other cytokines, to serum-free cultures restored missing CSF cell IgG secretion (ii) the inclusion of anti-IL-6, but not of control, blocking MoAb reduced IgG secretion by CSF cells in fetal calf serum (FCS)-containing cultures; and (iii) CSF cells were capable of active IL-6 production in the presence of FCS. These results suggest that endogenous IL-6 production by MS CSF cells seems to be responsible for inducing CSF IgG-secreting B cells to reach terminal differentiation.


Exploratory analysis of textual data from the Mother and Child Handbook using the text-mining method: Relationships with maternal traits and post-partum depression.
The aim of the present study was to examine the possibility of screening apprehensive pregnant women and mothers at risk for post-partum depression from an analysis of the textual data in the Mother and Child Handbook by using the text-mining method. Uncomplicated pregnant women (n = 58) were divided into two groups according to State-Trait Anxiety Inventory grade (high trait [group I, n = 21] and low trait [group II, n = 37]) or Edinburgh Postnatal Depression Scale score (high score [group III, n = 15] and low score [group IV, n = 43]). An exploratory analysis of the textual data from the Maternal and Child Handbook was conducted using the text-mining method with the Word Miner software program. A comparison of the 'structure elements' was made between the two groups. The number of structure elements extracted by separated words from text data was 20 004 and the number of structure elements with a threshold of 2 or more as an initial value was 1168. Fifteen key words related to maternal anxiety, and six key words related to post-partum depression were extracted. The text-mining method is useful for the exploratory analysis of textual data obtained from pregnant woman, and this screening method has been suggested to be useful for apprehensive pregnant women and mothers at risk for post-partum depression.


The Dynamics of Adaptation to Stress from Standing Genetic Variation and de novo Mutations.
Adaptation from standing genetic variation is an important process underlying evolution in natural populations, but we rarely get the opportunity to observe the dynamics of fitness and genomic changes in real time. Here, we used experimental evolution and Pool-Seq to track the phenotypic and genomic changes of genetically diverse asexual populations of the yeast Saccharomyces cerevisiae in four environments with different fitness costs. We found that populations rapidly and in parallel increased in fitness in stressful environments. In contrast, allele frequencies showed a range of trajectories, with some populations fixing all their ancestral variation in <30 generations and others maintaining diversity across hundreds of generations. We detected parallelism at the genomic level (involving genes, pathways, and aneuploidies) within and between environments, with idiosyncratic changes recurring in the environments with higher stress. In particular, we observed a tendency of becoming haploid-like in one environment, whereas the populations of another environment showed low overall parallelism driven by standing genetic variation despite high selective pressure. This work highlights the interplay between standing genetic variation and the influx of de novo mutations in populations adapting to a range of selective pressures with different underlying trait architectures, advancing our understanding of the constraints and drivers of adaptation.


Attenuated Salmonella carrying siRNA-PD-L1 and radiation combinatorial therapy induces tumor regression on HCC through T cell-mediated immuno-enhancement.
Hepatocellular carcinoma (HCC), the most prevalent type of aggressive liver cancer, accounts for the majority of liver cancer diagnoses and fatalities. Despite recent advancements in HCC treatment, it remains one of the deadliest cancers. Radiation therapy (RT) is among the locoregional therapy modalities employed to treat unresectable or medically inoperable HCC. However, radioresistance poses a significant challenge. It has been demonstrated that RT induced the upregulation of programmed death ligand 1 (PD-L1) on tumor cells, which may affect response to PD-1-based immunotherapy, providing a rationale for combining PD-1/PD-L1 inhibitors with radiation. Here, we utilized attenuated Salmonella as a carrier to explore whether attenuated Salmonella carrying siRNA-PD-L1 could effectively enhance the antitumor effect of radiotherapy on HCC-bearing mice. Our results showed that a combination of siRNA-PD-L1 and radiotherapy had a synergistic antitumor effect by inhibiting the expression of PD-L1 induced by radiation therapy. Mechanistic insights indicated that the combination treatment significantly suppressed tumor cell proliferation, promoted cell apoptosis, and stimulated immune cell infiltration and activation in tumor tissues. Additionally, the combination treatment increased the ratios of CD4+ T, CD8+ T, and NK cells from the spleen in tumor-bearing mice. This study presents a novel therapeutic strategy for HCC treatment, especially for patients with RT resistance.


Regular physical exercise improves endothelial function in heart transplant recipients.
Impaired endothelial function is detectable in heart transplant (HTX) recipients and regarded as risk factor for coronary artery disease. We have studied whether endothelial function can be improved in HTX patients participating in a regular physical training program as demonstrated in patients with chronic heart failure, hypertension and coronary artery disease. Male HTX patients and healthy, age-matched controls were studied. Seven HTX patients (age: 60 +/- 6 yr; 6 +/- 2 yr of HTX) participated in an outpatient training program, six HTX patients (age: 63 +/- 8 yr; 7 +/- 1 yr of HTX) maintained a sedentary lifestyle without regular physical exercise since transplantation. A healthy control group comprised six subjects (age: 62 +/- 6 yr). Vascular function was assessed by flow-mediated dilation of the brachial artery (FMD). Systemic haemodynamic responses to intravenous infusion of the endothelium independent vasodilator sodium nitroprusside (SNP) and to NG-monomethyl-L-arginine (L-NMMA), an inhibitor of constitutive nitric oxide synthase, were also measured. Resting heart rate was significantly lower (p < 0.05) in healthy controls (66 +/- 13) than in the HTX training group (83 +/- 11) and in non-training HTX patients (91 +/- 9), baseline blood pressure also tended to be lower in healthy subjects and in the training HTX patients. FMD was significantly higher (p < 0.05) in the control group (8.4 +/- 2.2%) and in the training group (7.1 +/- 2.4%), compared with non-training HTX patients (1.4 +/- 0.8%). The response of systolic blood pressure (p = 0.08) and heart rate (p < 0.05) to L-NMMA was reduced in sedentary HTX patients compared with healthy controls and heart rate response to SNP was also impaired in sedentary HTX patients. Regular aerobic physical training restores vascular function in HTX patients, who are at considerable risk for developing vascular complications. This effect is demonstrable in conduit and systemic resistance arteries.


2.45 GHz microwave irradiation-induced oxidative stress affects implantation or pregnancy in mice, Mus musculus.
The present experiment was designed to study the 2.45 GHz low-level microwave (MW) irradiation-induced stress response and its effect on implantation or pregnancy in female mice. Twelve-week-old mice were exposed to MW radiation (continuous wave for 2 h/day for 45 days, frequency 2.45 GHz, power density=0.033549 mW/cm(2), and specific absorption rate=0.023023 W/kg). At the end of a total of 45 days of exposure, mice were sacrificed, implantation sites were monitored, blood was processed to study stress parameters (hemoglobin, RBC and WBC count, and neutrophil/lymphocyte (N/L) ratio), the brain was processed for comet assay, and plasma was used for nitric oxide (NO), progesterone and estradiol estimation. Reactive oxygen species (ROS) and the activities of ROS-scavenging enzymes- superoxide dismutase, catalase, and glutathione peroxidase-were determined in the liver, kidney and ovary. We observed that implantation sites were affected significantly in MW-irradiated mice as compared to control. Further, in addition to a significant increase in ROS, hemoglobin (p<0.001), RBC and WBC counts (p<0.001), N/L ratio (p<0.01), DNA damage (p<0.001) in brain cells, and plasma estradiol concentration (p<0.05), a significant decrease was observed in NO level (p<0.05) and antioxidant enzyme activities of MW-exposed mice. Our findings led us to conclude that a low level of MW irradiation-induced oxidative stress not only suppresses implantation, but it may also lead to deformity of the embryo in case pregnancy continues. We also suggest that MW radiation-induced oxidative stress by increasing ROS production in the body may lead to DNA strand breakage in the brain cells and implantation failure/resorption or abnormal pregnancy in mice.


Characterization of a specific alpha-mannosidase involved in oligosaccharide processing in Saccharomyces cerevisiae.
Fractionation of a crude extract from Saccharomyces cerevisiae X-2180 on Sepharose 6B in the presence of 0.5% Triton X-100 resolves two enzyme fractions containing alpha-mannosidase activity. Fraction I which is excluded from the gel contains alpha-mannosidase activity toward both p-nitrophenyl-alpha-D-mannopyranoside and Man9GlcNAc oligosaccharide as substrates, whereas Fraction II which is included in the gel contains only oligosaccharide alpha-mannosidase activity. The latter enzyme is very specific and removes a single mannose residue from Man9GlcNAc, whereas the alpha-mannosidase activity of Fraction I removes several mannose residues from Man9GlcNAc oligosaccharide. High resolution 1H NMR analysis of the Man8GlcNAc formed from Man9GlcNAc in the presence of the alpha-mannosidase of Fraction II showed only a single isomer with the following structure: (see formula; see text) This specific enzyme is most probably involved in processing of oligosaccharide during biosynthesis of mannoproteins. The mannose analog of 1-deoxynojirimycin (50-500 microM), dideoxy-1,5-imino-D-mannitol, inhibits the oligosaccharide alpha-mannosidase activities of Fractions I and II to about the same extent, but has no effect on the nonspecific alpha-mannosidase which acts on p-nitrophenyl-alpha-D-mannopyranoside.


The quality of life of adolescents with menstrual problems.
To date, very few publications have examined the health related quality of life (HRQL) in the younger population with menstrual problems, despite their high prevalence in adolescent girls. We describe the health-related quality of life (HRQL) among adolescents with menstrual problems and identified factors that have an impact on it. The study was a questionnaire study (using PedsQL 4.0) of adolescents aged 13-18 referred to a tertiary gynecology center for menstrual problems between June 2009 and August 2010. One hundred eighty-four adolescents completed the questionnaires. The mean age was 15.10 ± 1.49 with the mean body mass index (BMI) of 22.83 ± 4.82 kg/m(2). The most common menstrual problems seen in the clinic were dysmenorrhea (38.6%) followed by heavy bleeding (33.6%), oligomenorrhea (19.6%), and amenorrhea (8.2%). The mean overall score was 70.40 ± 16.36 with 42.3% having a score below 1 standard deviation (SD) from the norms. Adolescents with dysmenorrhea had the poorest score in physical function, whereas those with amenorrhea had the lowest score in psychosocial function. Maternal parenting style, parental anxiety, adolescents' ill-health behavior, and BMI have been found to have impact on the girls' quality of life (QoL). Although menstrual problems are not life threatening, they can pose a significant impact on the quality of life of these patients. Identification of these impacts might lead to the recognition of potential services or education to improve this. Understanding the characteristics that predict QoL may help a clinician identify patients who are risk for poor QoL.


Effects of Muscular Strength Training on Oral Health and Quality of Life: Using Korean Panel Survey Data, a Cross-Sectional Study.
The purpose of this study is to confirm the importance of muscular strength exercise by confirming the relationship between strength exercise, oral health, and quality of life. Using the 2019 and 2021 of the Korean National Health and Nutrition Examination Survey (KNHANES), 6535 people were selected as subjects. Complex sampling analysis was applied to all analyses; 2267 people were in the muscular strength training group (MSG), and 5841 people were in the non-muscular strength training group (NMSG). A multi-sample linear regression analysis was conducted to confirm the effect of muscular strength training on oral health and quality of life. As a result of confirming the effect of muscular strength training on oral health status, problems with chewing decreased by 0.105, and problems with speaking decreased by 0.028 with MSG compared to NMSG. In addition, compared to NMSG, it was confirmed that MSG reduced chewing discomfort by 0.047, while self-perceived oral health improved by 0.0123. Finally, as a result of confirming the effect of muscular strength training on oral health and quality of life in Korean adults, there was a significant effect on quality of life despite adjusting for sociodemographic characteristics and oral-health-related factors (p < 0.05). In this study, the relationship between muscular strength training and quality of life was confirmed. Therefore, efforts should be made to make oral health management and muscular strength training a part of life in relation to quality of life.


A pure compound from Curcuma comosa Roxb. protects neurons against hydrogen peroxide-induced neurotoxicity via the activation of Nrf-2.
Oxidative stress plays a pivotal role in several human diseases including Parkinson's disease (PD). Curcuma comosa, a member of Zingiberaceae, is widely known in Thailand as an alternative medicinal herb for uterine inflammation and estrogenic properties. In this study (3S)-1-(3,4-dihydroxyphenyl)-7-phenyl-(6E)-6-hepten-3-ol or compound 092 (C-092, or ASPP 092), a pure compound isolated from ethanol extract of C. comosa, was evaluated for neuroprotective effect on hydrogen peroxide-induced toxicity in SH-SY5Y cells. C-092 demonstrated a radical scavenging effect with comparable efficacy to ascorbic acid and exhibited a neuroprotective effect via suppression of apoptotic cell death as evidenced by a reduction in phospho-p53 and cleaved caspase-3 expression. C-092 causes induction of Nrf-2, which is a transcription factor responsible for the expression of a range of antioxidant genes. Moreover, the reduction in catalase activity caused by hydrogen peroxide was also alleviated by C-092 treatment. These results suggested the therapeutic potential of this compound for neurodegenerative diseases caused by oxidative stress.


Naloxone-induced pulmonary edema.
We present the case of a 68-year-old woman with acute pulmonary edema secondary to the administration of naloxone to reverse an inadvertent narcotic overdose. The patient presented following a 12-hour history of increasingly bizarre behavior and confusion. A total IV dose of 1.6 mg naloxone was administered in an attempt to reverse the suspected overconsumption of a codeine-containing cough suppressant. She immediately became agitated, tachycardic, and diaphoretic; a clinical diagnosis of acute pulmonary edema was made. Following treatment with furosemide, nitroglycerin, and morphine sulfate, the patient recovered completely without further incident. Although naloxone is thought to be a safe drug with few complications, it should not be used indiscriminantly, and the smallest doses necessary to elicit the desired response should be used.


Evidence of dual sexual transmission of multi-resistant HIV with two years persistence of resistance to NRTI and NNRTI: a case report.
A case report of dual sexual transmission, with secondary transmission from naïve to naïve patient, of HIV harbouring K103N and L100I mutations, conferring full non-nucleoside reverse transcriptase inhibitor (NNRTI) resistance, plus 2 nucleoside analogous reverse transcriptase inhibitor (NRTI) mutations is described. The secondary transmission of the resistant virus was confirmed by phylogenetic analysis. Data also suggest that mutations related to NRTI and NNRTI resistance may persist for a long time in naive patients, over 2 years in the present case report.


Involvement of BREK, a serine/threonine kinase enriched in brain, in NGF signalling.
We identified AATYK2 (Apoptosis-Associated Tyrosine Kinase 2) through a database search as a kinase specifically expressed in the brain. After characterization, we renamed it BREK (Brain-Enriched Kinase). Mouse BREK mRNA is expressed predominantly in brain, especially in olfactory bulb, olfactory tubercle, hippocampus, striatum, cerebellum, and cerebral cortex. Levels of expression and phosphorylation of BREK were high at 0-2 weeks after birth, suggesting that BREK is involved in neural development and functions during the early postnatal period. Phosphoamino acid analysis following in vitro kinase reaction revealed that BREK is a catalytically active, serine/threonine kinase. In PC12 cells, BREK was phosphorylated rapidly upon stimulation with nerve growth factor (NGF) in a protein kinase C-dependent pathway. In differentiated PC12 cells, BREK was enriched in cell bodies and growth cones, and also present along neurites. Introduction of a kinase-defective mutant of BREK into PC12 cells enhanced both ERK phosphorylation and neurite outgrowth in response to NGF, suggesting that BREK is a negative regulator of NGF-induced neuronal differentiation. Thus, we conclude that BREK is a new member of the family of protein serine/threonine kinases and that it plays important roles in NGF-TrkA signalling.


mRNA Profiling for Vaginal Fluid and Menstrual Blood Identification.
Recently mRNA profiling has been widely proposed as a universal tool for biological fluids identification. Here, we describe a test for vaginal fluid identification that combines detection of five markers: vaginal mRNAs and Lactobacilli in end-point PCR reaction. The test detects the following transcripts: HBD1 (Human beta-defensin 1), MUC4 (Mucin 4), MMP11 (Matrix metalloproteinase 11), housekeeping gene G6PDH (glucose 6-phosphate dehydrogenase), and the 16S-23S rRNA intergenic spacer regions of L. crispatus and L. gasseri/L. johnsonii. Simultaneous analysis of five vaginal markers and a housekeeping gene ensures high specificity and reliability in the detection of vaginal material, which could not be obtained using detection of a single marker.


Cardiac reoperations in octogenarians: Do they really benefit?
According to the demographic development of our society, the numbers of octogenarians referred to cardiac surgery are continuously growing. Although the benefit of first-time cardiac procedures for these patients is well documented, the fate of octogenarians after redo-procedures, with special regard to long-term survival, functional status and quality of life, is poorly investigated. We retrospectively identified 84 consecutive patients aged ≥80 years, who underwent a cardiac reoperation at the department for Cardiothoracic Surgery in the Heart & Vessel Center Bad Bevensen between January 2007 and 2013. Demographic profiles as well as operative data were analyzed, and the patients were prospectively followed. Patient's functional status and quality of life were assessed with the Barthel Index, New York Heart Association class and the short form-12 questionnaire. The mean age of the study group (61 men, 23 women) was 81.9 ± 1.9 years. Most redo-procedures were carried out after primary coronary artery bypass grafting (65%), primary aortic valve replacement (21%) and primary mitral valve replacement (6%). The most frequent actual surgical procedures were combined coronary artery bypass grafting and aortic valve replacement (26%), isolated coronary artery bypass grafting (19%), and isolated aortic valve replacement (19%). The mean length of hospital stay was 17 ± 15 days. In-hospital mortality counted for 32.1%. During follow up (29 ± 20 months) a further 19.0% of the patients died. The Barthel Index of the survivors was 89 ± 17 and their mean New York Heart Association class was 2 ± 1. A total of 93% of the patients were living at home. Summary scores of physical and mental health of the short form-12 questionnaire equalled those of an age- and sex-matched normative population. Despite high perioperative mortality, results document a sustainable recovery of the survivors offering the prospect of a highly independent and satisfying life. Therefore, advanced age alone should not be a contraindication for redo cardiac interventions. Geriatr Gerontol Int 2016; 16: 1138-1144.


COVPRIG robustly predicts the overall survival of IDH wild-type glioblastoma and highlights METTL1+ neural-progenitor-like tumor cell in driving unfavorable outcome.
Accurately predicting the outcome of isocitrate dehydrogenase (IDH) wild-type glioblastoma (GBM) remains hitherto challenging. This study aims to Construct and Validate a Robust Prognostic Model for IDH wild-type GBM (COVPRIG) for the prediction of overall survival using a novel metric, gene-gene (G × G) interaction, and explore molecular and cellular underpinnings. Univariate and multivariate Cox regression of four independent trans-ethnic cohorts containing a total of 800 samples. Prediction efficacy was comprehensively evaluated and compared with previous models by a systematic literature review. The molecular underpinnings of COVPRIG were elucidated by integrated analysis of bulk-tumor and single-cell based datasets. Using a Cox-ph model-based method, six of the 93,961 G × G interactions were screened to form an optimal combination which, together with age, comprised the COVPRIG model. COVPRIG was designed for RNA-seq and microarray, respectively, and effectively identified patients at high risk of mortality. The predictive performance of COVPRIG was satisfactory, with area under the curve (AUC) ranging from 0.56 (CGGA693, RNA-seq, 6-month survival) to 0.79 (TCGA RNAseq, 18-month survival), which can be further validated by decision curves. Nomograms were constructed for individual risk prediction for RNA-seq and microarray-based cohorts, respectively. Besides, the prognostic significance of COVPRIG was also validated in GBM including the IDH mutant samples. Notably, COVPRIG was comprehensively evaluated and externally validated, and a systemic review disclosed that COVPRIG outperformed current validated models with an integrated discrimination improvement (IDI) of 6-16%. Moreover, integrative bioinformatics analysis predicted an essential role of METTL1+ neural-progenitor-like (NPC-like) malignant cell in driving unfavorable outcome. This study provided a powerful tool for the outcome prediction for IDH wild-type GBM, and preliminary molecular underpinnings for future research.


Effects of petroleum hydrocarbon solvents on alkaline phosphatase of rats.
After administration of benzene, gasoline, IOMEX or petroleum ether, liver alkaline phosphatase showed over 200% increase in activity whereas kidney alkaline phosphatase activity was depressed by 50%. The pattern of separation of isoenzymes of alkaline phosphatase in liver, kidney and serum indicated involvement of one of the isoenzymes. A simple method of detecting the incipent toxicity is described.


Expression of brain-derived neurotrophic factor protein in activated microglia of human immunodeficiency virus type 1 encephalitis.
The role of neurotrophic factors and their therapeutic potential have been investigated in various neurodegenerative disorders. In neurodegeneration associated with human immunodeficiency virus (HIV) infection, neuronal function and survival may be affected by abnormal neurotrophic regulation involving HIV-infected microglia and reactive astrocytes. To characterize the cellular localization of brain-derived neurotrophic factor (BDNF) and its high-affinity tyrosine kinase receptor, trkB, proteins in HIV-1 encephalitis, we examined post-mortem brains from patients with acquired immunodeficiency syndrome and brains from non-HIV-infected controls. Using double immunofluorescent confocal microscopy, we found that BDNF immunoreactivity was distributed in neocortical neuronal perikarya and neuritic processes, while in the striatum only neurites were BDNF-immunoreactive. Additionally, the striatum with HIV infection was characterized by BDNF immunoreactivity in infiltrating activated microglia/macrophages and multinucleated giant cells. Catalytic trkB receptor immunoreactivity was observed in neuronal perikarya in the neocortex and striatum, as well as in reactive astrocytes within HIV-infected regions. Our findings suggest that expression of BDNF by activated microglia in HIV-1 encephalitis may affect neuronal survival and astroglial response through corresponding trkB receptors.


Age-related neuronal vulnerability to brain ischemia: A potential target of gene therapy.
Brain infarction is one of the most important age-associated diseases. We have developed aged animal models for brain ischemia, and found the age-related neuronal vulnerability to brain ischemia. Investigation of that mechanism would lead to the effective treatment of brain infarction in the elder population. Recent advancement of gene transfer technique has provided strong tools for the neuronal and vascular biology. We described our recent approaches of gene transfer to blood vessels, including cerebral circulation, using adenoviral vectors. Cerebral blood vessels, atherosclerotic endothelium, and ischemic brain tissue are good targets of gene transfer. Development of these techniques would offer new therapeutic strategies for the age-related neuronal vulnerability and other age-associated diseases.


Achieving big with small: quantitative clinical pharmacology tools for drug development in pediatric rare diseases.
Pediatric populations represent a major fraction of rare diseases and compound the intrinsic challenges of pediatric drug development and drug development for rare diseases. The intertwined complexities of pediatric and rare disease populations impose unique challenges to clinical pharmacologists and require integration of novel clinical pharmacology and quantitative tools to overcome multiple hurdles during the discovery and development of new therapies. Drug development strategies for pediatric rare diseases continue to evolve to meet the inherent challenges and produce new medicines. Advances in quantitative clinical pharmacology research have been a key component in advancing pediatric rare disease research to accelerate drug development and inform regulatory decisions. This article will discuss the evolution of the regulatory landscape in pediatric rare diseases, the challenges encountered during the design of rare disease drug development programs and will highlight the use of innovative tools and potential solutions for future development programs.


Acute effects of phenoxybenzamine on alpha-adrenoceptor responses in vivo and in vitro: relation of in vivo pressor responses to the number of specific adrenoceptor binding sites.
Pressor responses to intravenous phenylephrine, an adrenoceptor agonist, and to the alpha 2-adrenoceptor-selective agonist guanabenz were examined in conscious rabbits 30 min after treatment with a range of doses of phenoxybenzamine (10(-5) to 5 mg/kg). The maximum number of specific prazosin- and clonidine-binding sites in the spleen of rabbits sacrificed 30 min after receiving phenoxybenzamine were measured using radioligand binding techniques. Treatment with phenoxybenzamine resulted in a dose-dependent reduction in the maximum pressor response to both phenylephrine and guanabenz, although phenoxybenzamine was a more potent antagonist at postsynaptic alpha 1- than at postsynaptic alpha 2-adrenoceptors. Phenoxybenzamine treatment also caused a dose-dependent reduction in specific [3H]prazosin and [3H]clonidine binding. The maximum in vivo pressor response to guanabenz was observed only when all specific clonidine-binding sites were present. There was a close correlation between in vitro receptor number and in vivo pressor responses for alpha 2-adrenoceptor stimulation but not for alpha 1-adrenoceptor-mediated responses. The maximum pressor response to phenylephrine could be obtained in rabbits in which the number of specific prazosin-binding sites was reduced by 60%. These experiments provide an approach to relating in vitro receptor number to in vivo responses.


Psychobiological effects of carbohydrate- and protein-rich meals in patients with seasonal affective disorder and normal controls.
Patients with seasonal affective disorder (SAD) frequently report carbohydrate craving and note that carbohydrate ingestion energizes them. Bright artificial light has been shown to reverse the symptoms of SAD, including carbohydrate craving. In this study, 16 depressed SAD patients and 16 matched controls were fed two different isocaloric meals, one rich in protein and one rich in carbohydrates, in a crossover design. Although their biochemical response in terms of plasma large neutral amino acid concentrations was identical, SAD patients reported activation following carbohydrate ingestion, whereas normal controls reported sedation. Marked ordering effects on psychological parameters were noted, suggesting that order should be taken into account as a methodological consideration in meal studies.


Cetuximab enhances TRAIL-induced gastric cancer cell apoptosis by promoting DISC formation in lipid rafts.
TRAIL is a member of the tumor necrosis factor family that selectively induces cancer cell apoptosis. However, gastric cancer cells are insensitive to TRAIL. Our and others studies showed that the inhibition of EGFR pathway activation could increase the sensitivity of TRAIL in cancer cells. But the detailed mechanism is not fully understood. In the present study, compared with TRAIL or cetuximab (an anti-EGFR monoclonal antibody) alone, treatment with the TRAIL/cetuximab combination significantly promoted death receptor 4 (DR4) clustering as well as the translocation of both DR4 and Fas-associated death domain-containing protein (FADD) into lipid rafts. This in turn resulted in caspase-8 cleavage and the formation of the death-inducing signaling complex (DISC) in these lipid rafts. Cholesterol-depletion with methyl-β-cyclodextrin partially prevented DR4 clustering and DISC formation, and thus partially reversed apoptosis induced by the TRAIL/cetuximab dual treatment. These results indicate that cetuximab increases TRAIL-induced gastric cancer cell apoptosis at least partially through the promotion of DISC formation in lipid rafts.


Induction of fatty liver by Coleus forskohlii extract through enhancement of de novo triglyceride synthesis in mice.
Coleus forskohlii extract (CFE), an herbal ingredient, is used for weight-loss products. CFE's alleged efficacy is attributed to forskolin. However, CFE has been shown to induce fatty liver in mice, with components other than forskolin playing a part in this effect. The present study addressed the underlying mechanism of CFE-induced fatty liver by analyzing changes in CFE-treated mice of lipid concentrations and of the levels of mRNAs encoding enzymes and transcription factors known to be related to fatty liver. Mice were fed a diet containing 0, 0.3 and 1% CFE for 2 weeks. CFE at 1% clearly induced fatty liver, as demonstrated by histological examination and confirmed by increases in triglyceride concentrations in liver. However, treated mice did not exhibit elevation in plasma levels of non-esterified fatty acids. Comprehensive analysis of liver mRNA levels revealed accumulation of multiple transcripts, including mRNAs encoding enzymes acetyl-CoA carboxylase and long-chain elongase; transcription factor peroxisome proliferator-activated receptor gamma (PPARγ); and lipid-droplet-associated fat-specific protein 27 (Fsp27). These findings suggest that the de novo synthesis and accumulation of triglyceride in the liver, through the enhanced expression of specific lipogenic mRNAs, is a major underlying mechanism of fatty liver induction by CFE.


Advancing Health Equity Through Equity-Centered Leadership Development with Interprofessional Healthcare Teams.
Events of spring 2020-the COVID19 pandemic and re-birth of a social justice movement-have thrown disparities in disease risk, morbidity, and mortality in sharp relief. In response, healthcare organizations have shifted attentions and resources towards equity, diversity, and inclusion (EDI) issues and initiatives like never before. Focused, proven equity-centered skill and mindset development is needed for healthcare professionals to operationalize these pledges and stated aims. This article highlights program evaluation results for this Clinical Scholars National Leadership Institute (CSNLI) specific to EDI. We will show that CSNLI imparts the valuable and essential skills to health professionals that are needed to realize health equity through organizational and system change. Initial cohort of 29 participants in CSNLI, engaging in the program over 3 years through in-person and distance-based learning offerings and activities. The CSNLI is a 3-year, intensive leadership program that centers EDI skill development across personal, interpersonal, organizational, and systems domains through its design, competencies, and curriculum. A robust evaluation following the Kirkpatrick Model offers analysis of four data collecting activities related to program participants' EDI learning, behavioral change, and results. Over the course of the program, participants made significant gains in competencies related to equity, diversity, and inclusion. Furthermore, participants demonstrated growth in behavior change and leadership activities in the areas of organizational and system change. Results demonstrate the need to center both leader and leadership development on equity, diversity, and inclusion curriculum to make real change in the US Healthcare System.


Association of Bleeding Severity With Mortality in Extended Thromboprophylaxis of Medically Ill Patients in the MAGELLAN and MARINER Trials.
Extended thromboprophylaxis has not been widely implemented in acutely ill medical patients because of bleeding concerns. The MAGELLAN (Multicenter, Randomized, Parallel Group Efficacy and Safety Study for the Prevention of Venous Thromboembolism in Hospitalized Medically Ill Patients Comparing Rivaroxaban With Enoxaparin) and MARINER (Medically Ill Patient Assessment of Rivaroxaban Versus Placebo in Reducing Post-Discharge Venous Thrombo-Embolism Risk) trials evaluated whether rivaroxaban compared with enoxaparin or placebo could prevent venous thromboembolism without increased bleeding. We hypothesized that patients with major bleeding but not those with nonmajor clinically relevant bleeding would be at an increased risk of all-cause mortality (ACM). We evaluated all bleeding events in patients taking at least 1 dose of study drug and their association with ACM in 4 mutually exclusive groups: (1) no bleeding, or first event was (2) nonmajor clinically relevant bleeding, (3) major bleeding, or (4) trivial bleeding. Using a Cox proportional hazards model adjusted for differences in baseline characteristics associated with ACM, we assessed the risk of ACM after such events. Compared with patients with no bleeding, the risk of ACM for patients with nonmajor clinically relevant bleeding was not increased in MARINER (hazard ratio, 0.43; P=0.235) but was increased in MAGELLAN (hazard ratio, 1.74; P=0.021). Major bleeding was associated with a higher incidence of ACM in both studies, whereas trivial bleeding was not associated with ACM in either study. Patients with major bleeding had an increased risk of ACM, whereas nonmajor clinically relevant bleeding was not consistently associated with an increased risk of death. These results inform the risk-benefit calculus of extended thromboprophylaxis in medically ill patients. URL: https://www. gov; Unique identifier: MAGELLAN, NCT00571649. URL: https://www. gov; Unique identifier: MARINER, NCT02111564.


Not all durations of preheart transplant mechanical ventilation portend inferior post-transplant survival in children.
Mechanical ventilation prior to pediatric heart transplantation predicts inferior post-transplant survival, but the impact of ventilation duration on survival is unclear. Data from the United Network for Organ Sharing and Pediatric Health Information System were used to identify pediatric (<18 years) heart transplant recipients from 2003 to 2020. Patients ventilated pretransplant were first compared to no ventilation, then ventilation durations were compared across quartiles of ventilation (≤1 week, 8 days-5 weeks, >5 weeks). At transplant, 11% (511/4506) of patients required ventilation. Ventilated patients were younger, had more congenital heart disease, more urgent listing-status, and greater rates of nephropathy, TPN-dependence, and inotrope and ECMO requirements (p < .001 for all). Post-transplant, previously ventilated patients experienced longer ventilation durations, ICU and hospital stays, and inferior survival (all p < .001). Hospital outcomes and survival worsened with longer pretransplant ventilation. One-year and overall survival were similar between the no-ventilation and ≤1 week groups (p = .703 & p = .433, respectively) but were significantly worse for ventilation durations >1 week (p < .001). On multivariable analysis, ventilation ≤1 week did not predict mortality (HR 0.98 [95% CI 0.85-1.43]), whereas ventilation >1 week did (HR: 1.18 [1.01-1.39]). Longer pretransplant ventilation portends worse outcomes, although only ventilation >1 week predicts mortality. These findings can inform pretransplant prognostication.


Structural elements of bactopterin from Pseudomonas carboxydoflava carbon monoxide dehydrogenase.
Bactopterin is a novel pterin occurring in bacterial molybdoenzymes as the organic portion of the molybdenum cofactor. Its structure is investigated here. The compound contains a single pterin ring and carries a side chain at carbon atom 6 of the pterin nucleus as indicated by the formation of pterin-6-carboxylic acid upon alkaline permanganate oxidation. Studies with phosphate-cleaving enzymes revealed the presence of two monophosphoric acid monoesters. The affinity of reduced bactopterin for thiol-Sepharose points to the presence of thiol(s) in active bactopterin.