Di(Thioether Sulfonate)-Substituted Quinolinedione as a Rapidly Dissoluble and Stable Electron Mediator and Its Application in Sensitive Biosensors.
The commonly required properties of diffusive electron mediators for point-of-care testing are rapid dissolubility, high stability, and moderate formal potential in aqueous solutions. Inspired by nature, various quinone-containing electron mediators have been developed; however, satisfying all these requirements remains a challenge. Herein, a strategic design toward quinones incorporating sulfonated thioether and nitrogen-containing heteroarene moieties as solubilizing, stabilizing, and formal potential-modulating groups is reported. A systematic investigation reveals that di(thioether sulfonate)-substituted quinoline-1,4-dione (QLS) and quinoxaline-1,4-dione (QXS) display water solubilities of ≈1 m and are rapidly dissoluble. By finely balancing the electron-donating effect of the thioethers and the electron-withdrawing effect of the nitrogen atom, formal potentials suitable for electrochemical biosensors are achieved with QLS and QXS (-0.15 and -0.09 V vs Ag/AgCl, respectively, at pH 7.4). QLS is stable for >1 d in PBS (pH 7.4) and for 1 h in tris buffer (pH 9.0), which is sufficient for point-of-care testing. Furthermore, QLS, with its high electron mediation ability, is successfully used in biosensors for sensitive detection of glucose and parathyroid hormone, demonstrating detection limits of ≈0.3 × 10-3 m and ≈2 pg mL-1 , respectively. This strategy produces organic electron mediators exhibiting rapid dissolution and high stability, and will find broad application beyond quinone-based biosensors.


The need for supplemental breast cancer screening modalities: a perspective of population-based breast cancer screening programs in Japan.
This article discusses possible supplemental breast cancer screening modalities for younger women with dense breasts from a perspective of population-based breast cancer screening program in Japan. Supplemental breast cancer screening modalities have been proposed to increase the sensitivity and detection rates of early stage breast cancer in women with dense breasts; however, there are no global guidelines that recommend the use of supplemental breast cancer screening modalities in such women. Also, no criterion standard exists for breast density assessment. Based on the current situation of breast imaging in Japan, the possible supplemental breast cancer screening modalities are ultrasonography, digital breast tomosynthesis, and breast magnetic resonance imaging. An appropriate population-based breast cancer screening program based on the balance between cost and benefit should be a high priority. Further research based on evidence-based medicine is encouraged. It is very important that the ethnicity, workforce, workflow, and resources for breast cancer screening in each country should be considered when considering supplemental breast cancer screening modalities for women with dense breasts.


Technical difficulties of removal of locking screw after locking compression plating.
As there are few reports on the difficulties of removing the locking compression plate (LCP), we prospectively investigated the incidence and difficulties in 58 patients in whom various types of LCPs were taken. From January 2004 to December 2007, we have removed 159 5.0-self tapping locking screws and 279 3.5- self tapping locking screws. All of the operations were performed by experienced trauma surgeons. All of the screws were inserted with the use of torque limiting attachment according to the manufacturer's recommendation. During the same period of time we have removed 198 AO-3.5 cortical and 4.0 cancellous screws from various sites. All of 159 5.0-self tapping locking screws were removed without difficulties. A total of 24 out of 279 3.5- self tapping locking screws were removed with many difficulties due to the stripping of the hexagonal recess. The use of conical extraction screw which was developed especially for the removal of stripped locking screws was successful in only six screws. We have removed plates by cutting the plate with metal cutting saw. We describe useful technical trick to remove the plate when there is only one screw left stripped. Compared to the locking screws, only one of 198 3.5-cortical screws was stripped. Care should be taken at the time of removal of the locking compression plate, especially for the 3.5-locking screws.


Ovarian yolk sac tumor arising from an endometrioid carcinoma.
A yolk sac tumor that arose within an ovarian endometrioid adenocarcinoma in a 50-year-old woman is described. The tumor had typical microscopic features, stained immunohistochemically for alpha-fetoprotein, and was associated with an elevated serum alpha-fetoprotein level when metastases appeared. This is the fourth case in which a yolk sac tumor has been reported to develop from a somatic carcinoma and the second in which this tumor has been associated with epithelium of endometrioid type. Tumor heterogeneity or neometaplasia may account for the origin of a tumor of germ cell type from a carcinoma of somatic origin.


Preliminary study: Taiwanese mothers' experiences of children with sensory processing disorder.
Different cultural backgrounds and parental experiences influence parenting styles and approaches to raising children with disabilities. Family-centered care should consider parental, especially main caregivers, experiences with their disabled children. In Taiwan, in most of homes, mothers are the main caregivers. The purpose of this study was to explore Taiwanese mothers' experiences with their children who have sensory processing disorder. This study used a qualitative ethnographic design and semistructured interview format. Transcripts were analyzed and synthesized into themes. Three mothers were interviewed. The following three themes emerged: (a) relationships within the shared worlds of disability and culture, (b) daily life challenges and expectations, and (c) opportunity to receive professional services. These Taiwanese mothers expressed that they experienced stress from being blamed for insufficient skills and from the shame of their children's disabilities that reflected lack of teacher, friend, and other family members' understanding of the cause of their children's inappropriate behaviors. Their children experienced difficulties performing daily activities, which resulted in stresses on both the mother and her child. The interviewed mothers needed to receive timely, long-term services from healthcare professionals. However, hospitals are inadequately staffed with occupational therapists, which delays care for children with special needs. Taiwanese mothers experience stresses from themselves, their child, and others. Healthcare professionals should apply a family-centered service approach to fulfill the needs of mothers and their disabled children. Moreover, healthcare professionals should promote greater awareness of sensory processing disorder symptoms and interventions to increase public awareness and acceptance of these children.


Use of the synbiotic VSL#3 and yacon-based concentrate attenuates intestinal damage and reduces the abundance of Candidatus Saccharimonas in a colitis-associated carcinogenesis model.
Individuals with inflammatory bowel disease are at high risk of developing colitis-associated cancer; thus, strategies to inhibit disease progression should be investigated. The study aimed to explore the role of the synbiotic (probiotic VSL#3® and yacon-based concentrate) in a colitis-associated carcinogenesis model. IL-10-/- mice were induced to carcinogenesis with 1,2-dimethylhydrazine and divided into two experimental groups: control and synbiotic. Manifestations of colitis, colon histology, expression of antioxidant enzymes, production of organic acids and intestinal microbiota were evaluated. The use of the synbiotic showed benefits, such as the preservation of intestinal architecture, increased expression of antioxidant enzymes and the concentration of organic acids, especially butyrate. It was also observed different microbial community profiles between the groups during the study. Together, these factors contributed to mitigate the manifestations of colitis and improve intestinal integrity, suggesting the potential benefit of the synbiotic in intestinal diseases.


Organ- and non-organ-specific auto-antibodies in children with hypopituitarism on growth hormone therapy.
Serum non-organ-specific antibodies (NOSA) against nuclear, mitochondrial (AMA), smooth muscle, liver/kidney microsomal (LKM), reticulin, ribosomal, and organ-specific antibodies (OSA) against pituitary gland, gonads (testis, ovary) adrenal cortex, thyroid (thyroglobulin and microsomal), pancreas islet cells, gastric parietal cells and intestinal epithelial cells were evaluated in 45 patients with hypopituitarism (mean age 12.4 +/- 4.0 years). In 22 of them, 9 with isolated growth hormone (GH) deficiency (IGHD) and 13 with multiple pituitary hormone deficiency, MRI showed anterior pituitary hypoplasia with structural stalk abnormality and ectopic posterior pituitary. Twelve had isolated small anterior pituitary and IGHD and 11 had normal morphology of pituitary gland and IGHD. Controls were healthy age-sex-matched subjects. Thyroid antibodies were detected by a passive haemagglutination test while indirect immunofluorescence was used for the others. The auto-antibodies were found in 7/45 asymptomatic patients, a frequency not significantly different from that in controls; 5 were type NOSA and 2 type OSA. Pituitary antibodies were positive in 1 girl with IGHD and normal pituitary morphology. One girl developed hyperthyroidism during the follow up. Autoantibodies were equally distributed between the three groups and the frequency was not dissimilar from that in controls; this suggests that these patients are not at a higher risk of developing auto-immune disease, at least during the first two decades. Pituitary insufficiency in children with different MRI features seems unlikely to be secondary to an auto-immune process.


Prototyping for public health in a local context: a streamlined evaluation of a community-based weight management programme (Momenta), Northumberland, UK.
Stakeholder co-production in design of public health programmes may reduce the 'implementation gap' but can be time-consuming and costly. Prototyping, iterative refining relevant to delivery context, offers a potential solution. This evaluation explored implementation and lessons learnt for a 12-week referral-based weight-management programme, 'Momenta', along with feasibility of an iterative prototyping evaluation framework. Mixed methods evaluation: Qualitative implementation exploration with referrers and service users; preliminary analysis of anonymised quantitative service data (12 and 52 weeks). Two leisure centres in Northumberland, North East England. Individual interviews with referring professionals (n=5) and focus groups with service users (n=13). Individuals (n=182) referred by healthcare professionals (quantitative data). Three 12-week programme iterations: Momenta (n=59), Momenta-Fitness membership (n=58) and Fitness membership only (n=65). Primary outcome: Qualitative themes developed through stakeholder-engagement. Secondary outcomes included preliminary exploration of recruitment, uptake, retention, and changes in weight, body mass index, waist circumference and psychological well-being. Service users reported positive experiences of Momenta. Implementation gaps were revealed around the referral process and practitioner knowledge. Prototyping enabled iterative refinements such as broadening inclusion criteria. Uptake and 12-week retention were higher for Momenta (84.7%, 45.8%) and Momenta-Fitness (93.1%, 60.3%) versus Fitness only (75.4%, 24.6%). Exploration of other preliminary outcomes (completers only) suggested potential for within-group weight loss and increased psychological well-being for Momenta and Momenta-Fitness at 12 weeks. 52 week follow-up data were limited (32%, 33% and 6% retention for those who started Momenta, Momenta-Fitness and Fitness, respectively) but suggested potential weight loss maintenance for Momenta-Fitness. Identification of issues within the referral process enabled real-time iterative refinement, while lessons learnt may be of value for local implementation of 'off-the-shelf' weight management packages more generally. Our preliminary data for completers suggest Momenta may have potential for weight loss, particularly when offered with a fitness membership.


Abnormal origin of the posterior inferior cerebellar artery. Case report.
Posterior inferior cerebellar arteries originating from the vertebral arteries between the axis and the atlas were demonstrated angiographically in a 12-year-old boy. The patient presented with tetraparesis of sudden onset after a gymnastic exercise. There was no recognizable trauma.


Optimization of extraction and dyeing conditions of the Parkia roxburghii pod's peel as a natural substantive silk dye.
Natural dyes are soft, eco-friendly, and non-toxic colours for silk fabric, imparting high compatibility with the environment. Among the different natural dyestuffs derived from different parts of plants, the peel of the Parkia roxburghii pod is thriving as one of the potential natural substantive dyes. The study reveals optimization of dye extraction for dyeing silk fabric. Colour strength (K/S) and absorbance of dye extract were evaluated for optimization of extraction and dyeing parameters. The optimized ML ratio (material and solvent proportions) was 1:30 for 60 min of boiling at 80 °C under acidic medium. Use of natural and synthetic mordants produced different colour patterns giving two classes, YR, light to dark brownish colour. CuSO4, lime juice and Terminalia chebula mordants produced better wash fastness and light fastness in meta-mordant conditions. Dyeing silk with parkia peel without mordant also produces better fastness properties, thus acts as a natural substantive silk dye.


Prevalence of MS in south Estonia. Evidence of a new border of the Fennoscandian focus.
A population-based study of MS was carried out in South Estonia in 1988-1989. Cases were identified from the Tartu University Hospital archives where all MS cases in South Estonia are diagnosed, from all neurologists and nursing homes of the region, and from the local Multiple Sclerosis Society. The prevalence in South Estonia is 51 per 100,000, for native Estonians 55, for Russians 29, for other nationalities 42 per 100,000. The prevalence rate in different counties was demonstrated as low as 31 per 100,000 in the County of Tartu, to 72 per 100,000 in Pôlva County; 55% of patients have retired because of their handicap and only 2 patients (1%) were living in nursing homes.


T lymphocyte lines producing or vaccinating against autoimmune encephalomyelitis (EAE). Functional activation induces peanut agglutinin receptors and accumulation in the brain and thymus of line cells.
We studied lines of rat T cells, specifically reactive against myelin basic protein (BP), that were functional in mediating autoimmune encephalomyelitis or in vaccinating rats against induction of active EAE. Herein we report that these functions depended on activation of the cells by incubation with BP or with a T cell mitogen prior to inoculation into recipient rats. Activation was accompanied by the exposure of membrane-binding sites specific for the lectin peanut agglutinin. Accumulation of activated line cells in the central nervous system and thymus gland was observed.


Iron storage in bacteria.
Iron is an essential nutrient for nearly all organisms but presents problems of toxicity, poor solubility and low availability. These problems are alleviated through the use of iron-storage proteins. Bacteria possess two types of iron-storage protein, the haem-containing bacterioferritins and the haem-free ferritins. These proteins are widespread in bacteria, with at least 39 examples known so far in eubacteria and archaebacteria. The bacterioferritins and ferritins are distantly related but retain similar structural and functional properties. Both are composed of 24 identical or similar subunits (approximately 19 kDa) that form a roughly spherical protein (approximately 450 kDa, approximately 120 A diameter) containing a large hollow centre (approximately 80 A diameter). The hollow centre acts as an iron-storage cavity with the capacity to accommodate at least 2000 iron atoms in the form of a ferric-hydroxyphosphate core. Each subunit contains a four-helix bundle which carries the active site or ferroxidase centre of the protein. The ferroxidase centres endow ferrous-iron-oxidizing activity and are able to form a di-iron species that is an intermediate in the iron uptake, oxidation and core formation process. Bacterioferritins contain up to 12 protoporphyrin IX haem groups located at the two-fold interfaces between pairs of two-fold related subunits. The role of the haem is unknown, although it may be involved in mediating iron-core reduction and iron release. Some bacterioferritins are composed of two subunit types, one conferring haem-binding ability (alpha) and the other (beta) bestowing ferroxidase activity. Bacterioferritin genes are often adjacent to genes encoding a small [2Fe-2S]-ferredoxin (bacterioferritin-associated ferredoxin or Bfd). Bfd may directly interact with bacterioferritin and could be involved in releasing iron from (or delivering iron to) bacterioferritin or other iron complexes. Some bacteria contain two bacterioferritin subunits, or two ferritin subunits, that in most cases co-assemble. Others possess both a bacterioferritin and a ferritin, while some appear to lack any type of iron-storage protein. The reason for these differences is not understood. Studies on ferritin mutants have shown that ferritin enhances growth during iron starvation and is also involved in iron accumulation in the stationary phase of growth. The ferritin of Campylobacter jejuni is involved in redox stress resistance, although this does not appear to be the case for Escherichia coli ferritin (FtnA). No phenotype has been determined for E. coli bacterioferritin mutants and the precise role of bacterioferritin in E. coli remains uncertain.


PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates.
A target length limitation to PCR amplification of DNA has been identified and addressed. Concomitantly, the base-pair fidelity, the ability to use PCR products as primers, and the maximum yield of target fragment were increased. These improvements were achieved by the combination of a high level of an exonuclease-free, N-terminal deletion mutant of Taq DNA polymerase, Klentaq1, with a very low level of a thermostable DNA polymerase exhibiting a 3'-exonuclease activity (Pfu, Vent, or Deep Vent). At least 35 kb can be amplified to high yields from 1 ng of lambda DNA template.


Molecular virology of chronic hepatitis B and C: parallels, contrasts and impact on drug development and treatment outcome.
Chronic infections with hepatitis B virus (HBV) and hepatitis C virus (HCV) are highly prevalent worldwide, causing significant liver disease and thus representing high unmet medical needs. Accordingly, substantial pharmaceutical and clinical research efforts have been made to develop and improve treatments for these viruses. While HBV and HCV are both hepatotropic viruses that can cause similar disease in chronically infected patients, they belong to different viral families. There are substantial differences in the molecular virology of HBV and HCV that have profound implications for therapeutic strategy. In particular, HBV has a long-lived nuclear form of its genome (covalently closed circular DNA) that is able to persist in the face of potent inhibition of viral replication. In contrast, HCV does not have a long-lived genome form and depends on active replication to maintain infection; HCV is therefore much more susceptible to eradication by potent antiviral agents. Additional differences between HBV and HCV with therapeutic implications include the size, structure and heterogeneity of their respective viral genomes. These factors influence the number of targets available for therapeutic intervention, response to therapy among viral genotypes and the emergence of viral resistance. Substantial progress has been made in treating each infection, but unique challenges remain. In this review, key differences in the molecular virology of hepatitis B and C will be presented, highlighting their impact on antiviral therapy (particularly with respect to direct-acting antivirals) and the challenges they present to the cure of each disease.


Role of MRI in the early diagnosis of tubal ectopic pregnancy.
To determine the role of MRI in the early diagnosis of tubal ectopic pregnancy (EP). Clinical and MRI features of 27 cases of tubal pregnancy were reviewed. A thick-walled gestational sac (GS)-like structure was demonstrated lateral to the uterus in all cases. On T2-weighted images, the thick wall typically exhibited 3 discrete rings in 22 cases (81 %), among which 17 cases (63 %) displayed small vessels and 6 cases (33 %) exhibited small areas of fresh haemorrhage inside the thick wall. The contents demonstrated non-specific liquid in 26 %, papillary solid components in 56 %, and fresh blood or fluid-fluid level in 19 % of the cases. Dilatation of the affected fallopian tube associated with hematosalpinx was demonstrated in 18 cases (67 %) and marked enhancement of the tubal wall was observed in 22 cases (81 %). No correlation was found between the size of the GS and the estimated gestational age (r = 0.056). MRI plays an important role in the early diagnosis and management of tubal pregnancy. The characteristic MRI features include a GS-like structure with a "three rings" appearance on T2-weighted images, presence of solid components in the sac, dilatation of the affected fallopian tube with hematosalpinx, and tubal wall enhancement. • MR imaging has served as a problem-solving procedure in ectopic pregnancy. • MR imaging features can be criteria for early diagnosis of tubal pregnancy. • Detailed assessment of ectopic implantation is necessary for management decision-making.


Transferrin receptor-1 and 2 expression in chronic lymphocytic leukemia.
Transferrin receptor (TfR)-1 and 2 mRNA and CD71 (TfR1) expression was analyzed in 118 CLL patients. Ninety-five out of 109 analyzed cases expressed CD71, mostly at a high level; 60% of CD71 (+) cases were IGH-mutated. All samples were TfR1 mRNA (+); TfR2-alpha/beta mRNA was detected in, respectively, 52/102 and 100/109 cases. Competitive RT-PCR showed widely divergent levels of TfR1 mRNA in cases with high CD71 expression, alluding to post-transcriptional control of TfR1 expression in CLL. The almost uniformly high CD71 expression in CLL is in keeping with the activated status of neoplastic cells, regardless of IGH mutational load.


Identification of operator sites within the upstream region of the putative mce2R gene from mycobacteria.
Mycobacterium tuberculosis harbors four mce operons. Among them, mce2 operon is preceded by a FadR-like regulator mce2R (Rv0586). Here, we report the operator sites of the mce2R and its orthologs in other sequenced mycobacteria and non-mycobacterial species Nocardia farciana. All the identified DNA motifs illustrate the FadR subfamily specific nucleotide preference. Moreover, these motifs from the upstream region share sequence conservation, which is in agreement with the similarity of their DNA binding domain. Using electrophoretic mobility shift assay, we demonstrate that the predicted DNA motifs specifically interact with the recombinant Mce2R-Rv0586. Our present study has implications in the understanding of cis-regulatory elements and the auto-regulatory nature of the FadR subfamily of regulators.


A radiographic analysis of anterior-posterior translation in total knee arthroplasty.
We performed radiological analyses to examine the relationship between the knee flexion angle and the anteroposterior translation movement relative to the prosthetic components (NexGen type) after total knee arthroplasty (TKA). Cruciate-retaining (CR) type TKA was performed in 12 knees with osteoarthritis (OA) and 9 knees with rheumatoid arthritis (RA) in which no posterior cruciate ligament (PCL) tear was present. Posterior-stabilized (PS) type TKA was performed in 7 OA knees and 7 RA knees in which the PCL was defective or resected. The measurements were performed according to the methods of Watanabe. The contact point ratio (percentage) was calculated by dividing the distance to the contact point (CP; the closest point of contact between the femoral and tibial components) by the antero-posterior length of the tibial component. After TKA, the CP at full extension was positioned more posteriorly than in the normal knee both under weight-bearing and non-weight-bearing conditions. Except for the RA knees in the PS group, the CP translated anteriorly in the early phase of flexion and then posteriorly. Under weight-bearing conditions, the posterior translation occurred earlier and was smaller in magnitude. For RA knees in the PS group, the CP moved gently posteriorly right from the beginning.


Fever-Induced Brugada-Pattern Electrocardiogram.
Brugada syndrome is an increasingly recognized syndrome characterized by a particular electrocardiography (ECG) pattern and clinical criteria and has a high incidence of sudden death in patients with structurally normal hearts. The Brugada ECG pattern can be unmasked by drugs, ischemia, and fever. We present the case of a 47-year-old man who presented to the emergency department with flu-like symptoms and syncope. On arrival, he was febrile and his ECG showed a Brugada pattern. Although this pattern resolved once his fever resolved, the cardiologists were concerned that his syncopal episode might have been due to ventricular tachycardia/fibrillation, and the patient was admitted for implantable cardiac defibrillator placement. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Fever and other stressors can unmask a Brugada pattern on ECG, and if patients have concerning clinical criteria, they should receive emergent cardiology follow-up.


Bivalirudin versus heparin plus a glycoprotein IIb/IIIa inhibitor in patients with non-ST-segment elevation myocardial infarction undergoing percutaneous coronary intervention after clopidogrel pretreatment: pooled analysis from the ACUITY and ISAR-REACT 4 trials.
The optimal antithrombotic therapy for patients with non-ST-segment elevation myocardial infarction undergoing percutaneous coronary intervention is not well defined. We investigated the efficacy and safety of bivalirudin versus heparin plus a glycoprotein IIb/IIIa inhibitor (GPI) in patients with non-ST-segment elevation myocardial infarction undergoing percutaneous coronary intervention after clopidogrel pretreatment. This study included 3798 clopidogrel-pretreated patients with non-ST-segment elevation myocardial infarction undergoing percutaneous coronary intervention, who were randomly assigned to receive bivalirudin (n=1928) or heparin (unfractionated heparin or enoxaparin; n=1870) plus a GPI in the setting of the Acute Catheterization and Urgent Intervention Triage Strategy (ACUITY) and Intracoronary Stenting and Antithrombotic Regimen: Rapid Early Action for Coronary Treatment (ISAR-REACT) 4 trials. Major end points were a composite of death, recurrent myocardial infarction or urgent target vessel revascularization (efficacy end point), major bleeding (safety end point), and the composite of death, recurrent myocardial infarction, urgent target vessel revascularization, or major bleeding (net adverse clinical events [NACE]) at 30 days. The incidence of the efficacy end point was 10.6% (n=205) in the bivalirudin group versus 10.2% (n=191) in the heparin plus a GPI group (OR, 1.04; 95% CI, 0.85-1.27; P=0.69). The incidence of safety end point was 3.4% (n=66) in the bivalirudin group versus 6.3% (n=117) in the heparin plus a GPI group (OR, 0.54 [0.40-0.72]; P<0.001). NACE occurred in 258 patients (13.4%) in the bivalirudin group versus 275 patients (14.7%) in the heparin plus a GPI group (OR, 0.90 [0.76-1.06]; P=0.21). NACE rates were not significantly different between bivalirudin and heparin plus a GPI in patients with non-ST-segment elevation myocardial infarction undergoing percutaneous coronary intervention after clopidogrel pretreatment. Although no significant difference in efficacy was seen in terms of suppression of adverse ischemic events, bivalirudin was superior to heparin plus a GPI in terms of reducing bleeding events. URL: http://www.clinicaltrials.gov. Unique Identifier: NCT00093158 and NCT00373451.


Coating with polysaccharides influences the surface charge of cerium oxide nanoparticles and their effects to Mytilus galloprovincialis.
This study focused on the effects of surface coating, acquired through the interaction with natural biomolecules, on the behavior and ecotoxicity of nanoparticles (NPs). To this aim, the effects of Cerium Oxide Nanoparticles (CeO2NPs) naked and coated with chitosan and alginate on the marine mussel Mytilus galloprovincialis were compared. Mussels were exposed for 7 days to 100 μg L-1 of CeO2NPs and for 28 days to 1 μg L-1 of CeO2NPs. In both experiments CeO2NPs were used naked and coated with the two polysaccharides. The lowest tested concentration allowed to understand the environmental relevance of this biological process. A set of biomarkers related to oxidative stress/damage and energy metabolism was applied to assess the ecotoxicity of CeO2NPs. The aggregation and stability in water of CeO2NPs were measured through dynamic light scattering analysis and the levels of Ce in the exposure media and in mussels soft tissues were determined by inductively coupled plasma-mass spectrometry. Results showed a different hydrodynamic behavior and stability of CeO2NPs in saltwater related to the different coatings. Despite this, no differences in the bioaccumulation of CeO2NPs were observed among the experimental groups. Different coatings affected also CeO2NPs toxicological outcomes in both 7- and 28-days exposures. Coating with chitosan enhanced antioxidant enzyme activities while coating with alginate triggered oxidative damage. Although the oxidant pathways did not differ that much among the exposures, biomarkers of energetic supplies suggested a different strategy of defense in response to CeO2NP exposure at a lower concentration and for a longer period of time. The obtained results are in line with findings of a previous study on freshwater mussels, suggesting that the coating with biomolecules, which impart negative charge to the NPs, might enhance their biological effects. This study highlighted that interactions of NPs with natural biomolecules largely present in the aquatic environment could affect NPs toxicity altering the interaction towards organisms.


Modelling the resource implications and budget impact of new reimbursement guidelines for the management of cow milk allergy in Finland.
To assess the impact of the decision by Kela (The Social Insurance Institution) to adopt guidelines for the management of food allergies in children, developed by the Finnish Medical Society Duodecim, as the basis for reimbursing clinical nutrition preparations for the treatment of cow milk allergy (CMA) in Finland. A decision model was constructed using published clinical outcomes and clinician-derived resource utilisation estimates. The model was used to estimate the net resource implications and associated costs of Kela's new policy for the annual cohort of 1443 new CMA sufferers over the first 6 months following referral to a specialist. The analysis was conducted from the perspective of Kela, patients and Finnish society. Kela's new policy for reimbursing clinical nutrition preparations for the treatment of CMA is expected to increase healthcare resource use, including a 10% increase in the number of specialist visits over the first 6 months following referral. Consequently, Kela's 6-monthly expenditure on 1443 new CMA sufferers following referral is expected to increase by 12% from 889,389 euros to 992,761 euros. Additionally, parents' costs are expected to increase by 10% and fathers' absenteeism from work to increase by 11% within the first 6 months following referral. It is important to validate guidelines in clinical practice before their implementation. Within the limitations of our model, Kela's new criteria for reimbursing clinical nutrition preparations for the treatment of CMA is expected to increase healthcare resource use in paediatric departments in public hospitals in Finland and increase costs to Kela, patients and Finnish society.


Luteinizing hormone induces ovulation via tumor necrosis factor α-dependent increases in prostaglandin F2α in a nonmammalian vertebrate.
Ovulation is induced by the preovulatory surge of luteinizing hormone (LH) that acts on the ovary and triggers the rupture of the preovulatory ovarian follicle by stimulating proteolysis and apoptosis in the follicle wall, causing the release of the mature oocyte. The pro-inflammatory cytokine tumor necrosis factor α (TNFα) and prostaglandin (PG) F2α (PGF2α) are involved in the control of ovulation but their role mediating the pro-ovulatory actions of LH is not well established. Here we show that Lh induces PGF2α synthesis through its stimulation of Tnfα production in trout, a primitive teleost fish. Recombinant trout Tnfα (rTnfα) and PGF2α recapitulate the stimulatory in vitro effects of salmon Lh (sLh) on contraction, proteolysis and loss of cell viability in the preovulatory follicle wall and, finally, ovulation. Furthermore, all pro-ovulatory actions of sLh are blocked by inhibition of Tnfα secretion or PG synthesis and all actions of rTnfα are blocked by PG synthesis inhibitors. Therefore, we provide evidence that the Tnfα-dependent increase in PGF2α production is necessary for the pro-ovulatory actions of Lh. The results from this study shed light onto the mechanisms underlying the pro-ovulatory actions of LH in vertebrates and may prove important in clinical assessments of female infertility.


Epigenetics of bladder cancer.
Epigenetic changes are closely associated with tumor development and prognosis of bladder cancer, and its detection in specimens (tissue/body fluid) may be feasible and more sensitive than conventional methods. Additionally, the epigenetic changes are interesting from a clinical point of view because it may be possible to reverse epigenetic changes and restore the gene function. Epigenetic markers might therefore be more useful than conventional molecular markers for the detection, prediction of prognosis, and treatment of bladder cancer.


Intraoperative ventilatory leak: Real-time guidance for management of air leak in lung cancer patients undergoing VATS lobectomy.
Persistent air leak (PAL) is a common complication after video-assisted thoracoscopic surgery (VATS) lobectomy. We aimed to evaluate whether the intraoperative quantitative measurement of air leaks using a mechanical ventilation test could predict PAL and identify those patients needing additional treatment for the prevention of PAL. This was an observational, retrospective, single-center study that included 82 patients who underwent VATS lobectomy with a mechanical ventilation test for VL. Only 2% of patients who underwent lobectomy surgery had persistent air leaks. At the end of lobectomy performed in patients with non-small cell lung cancer, the lung was reinflated at a 25-30 mmH2O pressure and ventilatory leaks (VL) were calculated and in relation to the entity of the air leaks, we evaluated the most suitable intraoperative treatment to prevent persistent air leaks. VL is an independent predictor of PAL after VATS lobectomy; it provides a real-time intraoperative guidance to identify those patients who can benefit from additional intraoperative preventive interventions to reduce PAL.


Enzymatic sulfation of coelenterazine by human cytosolic aryl sulfotransferase SULT1A1: identification of coelenterazine C2-benzyl monosulfate by LC/ESI-TOF-MS.
Coelenterazine (CTZ) is known as a light-emitting source for the bioluminescence reaction in marine organisms. CTZ has two phenolic hydroxy groups at the C2-benzyl and C6-phenyl positions, and a keto-enol type hydroxy group at the C3-position in the core structure of imidazopyrazinone (= 3,7-dihydroimidazopyrazin-3-one). These hydroxy groups in CTZ could be sulfated by sulfotransferase(s), and the sulfates of Watasenia luciferin (CTZ disulfate at the C2- and C6-positions) and Renilla pre-luciferin (CTZ 3-enol sulfate) have been identified in marine organisms. To characterize the sulfation process of CTZ, human cytosolic aryl sulfotransferase SULT1A1 (SUTase) was used as a model enzyme. The sulfated products catalyzed by SUTase with 3'-phosphoadenosine 5'-phosphosulfate (PAPS) were analyzed by LC/ESI-TOF-MS. The product was the monosulfate of CTZ and identified as the C2-benzyl sulfate of CTZ (CTZ C2-benzyl monosulfate), but CTZ disulfate, CTZ 3-enol sulfate, and CTZ C6-phenyl monosulfate were not detected. The non-enzymatic oxidation products of dehydrocoelenterazine (dCTZ, dehydrogenated derivative of CTZ), coelenteramide (CTMD), and coelenteramine (CTM) from CTZ were also identified as their monosulfates.


GDAP2 Overexpression Affects the Development of Neurons and Dysregulates Neuronal Excitatory Synaptic Transmission.
GDAP2 is a gene highly expressed in the human brain and encodes ganglioside-induced differentiation-associated protein 2 (GDAP2). At present, little is known about the function of GDAP2. In recent years, it has been reported that mutations in the GDAP2 gene may be involved in hereditary cerebellar ataxia. In this study, we first conducted a preliminary study on the effect of GDAP2 overexpression on cultured primary hippocampal neurons in vitro. By analysing neuronal morphology, it was found that the complexity of neurons and the number of dendritic spines increased when GDAP2 was upregulated. The electrophysiological recordings showed that GDAP2 overexpression significantly increased the frequency of mEPSCs, suggesting that GDAP2 overexpression dysregulates excitatory synaptic transmission in cultured primary hippocampal neurons in vitro. On the other hand, behavioural and field-potential recordings of epileptic mouse models showed that GDAP2 overexpression was associated with increased seizure frequency. In summary, this preliminary study suggested that GDAP2 overexpression may have a certain pathogenic effect, providing a new perspective for the study of gene-related diseases such as epilepsy.


High resolution scanning electron microscopy of frog sartorius muscle.
A field emission-type scanning electron microscope (SEM) was used to study the three-dimensional ultrastructure of frog sartorius muscles. Various preparative procedures were tested to seeks better specimen preparation for high resolution SEM observation. Procedures should be chosen depending on the information desired. The cell surface and intracellular organization of muscle fibers were best visualized when the tissues were fixed with tannic acid-OsO4 and torn after critical point drying. The basal lamina appeared as a continuous felt-like layer, onto which fine filamentous materials adhered. The true outer surface of the sarcolemma was not seen, whereas the true inner surface was occasionally exposed and exhibited numerous caveolae, membraneous fragments and fine filaments attached to its surface. In freeze-fractured and dried tissues, the cleaved sarcolemma showed numerous apertures of caveolae and T-system tubules. Inside the cell, the myofibrils showed a typical branding pattern of the sarcomere. Thick myofilaments were regularly beaded except for the pseudo-H-zone. Around the myofibrils the sarcoplasmic reticulum and T-system were also clearly observed. The results are discussed with special reference to the usefulness and limitation of the high resolution SEM in studying the ultrastructure of cells and tissues.


Growth in young rats after termination of sodium selenite exposure: studies of growth hormone and somatomedin C.
In a previous study we have shown that the growth retarding effect of selenium in infant rats occurs concomitantly with severely reduced growth hormone responses and serum somatomedin C. Furthermore, normal growth may be restored by administration of exogenous growth hormone administration. In the present study we examined the alterations after withdrawal of selenite administration in growth rate, serum growth hormone after GRF 40 injection and serum somatomedin C. Thirty-six rats were divided into 3 groups; i.e. controls, rats on continuous selenite treatment from days 21 to 63 of age and a group where selenium was withdrawn on day 42. The first 2 groups grew linearly during the experimental period, the second at a very significantly lower rate. In the third group after cessation of exposure a brief growth spurt occurred followed by a growth rate significantly different from that of the other 2 groups. The reduced GH response to GRF 40 stimulation was normalized 3 weeks after withdrawal, whereas serum somatomedin C remained at the same low level as in those rats on continuous selenite administration. There were other signs of liver lesion at day 42, i.e. elevated ALAT and reduced serum albumin, changes which became insignificant in all 3 groups at the end of the experiment. These results demonstrate that whereas stimulated growth hormone secretion was reestablished 3 weeks after termination of selenium exposure, severely reduced liver somatomedin C production persisted. An improvement in growth rate was also noted, but normal growth rate was not reached 3 weeks after discontinuation of the selenite treatment.


An improved operating room demagnetizer.
An operating room demagnetizer that utilized a pulsed, smoothly decaying oscillating magnetic field is described. The instrument can demagnetize a number of surgical instruments at one time and can provide instantaneous and complete demagnetization.


The rhetoric of specialization. The 1978 Harvey Cushing oration.
The 1978 Cushing Orator shows the role of rhetoric in the process by which various specialties change in response to sociological and legislative demands. He discusses the effect of applying concepts of restraint of trade to medicine, and welcomes a closer liaison through the American College of Surgeons between general surgeons and other surgical specialties.


Spotted fever group Rickettsia in brown dog ticks Rhipicephalus sanguineus in southwestern Spain.
A total of 2,229 adults ticks (1,428 males and 801 females) belonging to the brown dog tick, Rhipicephalus sanguineus Latreille, 1806, collected from dogs in Seville province (Andalusia), distributed in 500 lots ranging from one to eight specimens per lot, were examined for the presence of rickettsiae by molecular techniques. Specific rickettsiae DNA were detected in 90 lots (18%) of ticks tested. Sequence analysis of amplicons revealed that R. sanguineus ticks were infected exclusively with Rickettsia massiliae (including the strain Bar-29). The results of this study extend the knowledge of the geographic distribution and prevalence of these spotted fever group (SFG) rickettsiae and indicate that at least two of them, with yet uncertain pathogenicity to humans, are present in brown dog ticks in south western Spain. Although Mediterranean spotted fever (MSF) is an endemic disease in Andalusia, Rickettsia conorii was not found, whereas R. massiliae, recently described as a pathogenic species, was highly prevalent in this area. Our data suggest that in Andalusia a number of MSF or MSF-like cases attributed to R. conorii could have been actually caused by other SFG rickettsia present in R. sanguineus, particularly, R. massiliae.


Resistance to Boscalid in Botrytis cinerea From Greenhouse-Grown Tomato.
Gray mold, caused by the fungus Botrytis cinerea Pers ex Fr., is one of the most destructive spoilage diseases, severely affecting tomato production in Henan Province, China. Spraying fungicides from the flowering to the harvest stage is a necessary measure to reduce losses associated with B. cinerea infection. However, B. cinerea has developed resistance to fungicides in many countries. Boscalid is a succinate dehydrogenase inhibitor (SDHI) fungicide and was registered for the control of gray mold. In this study, a total of 269 B. cinerea isolates were collected from tomato in commercial greenhouses in different locations of Henan Province in 2014 and 2015. The sensitivity and resistance of B. cinerea field isolates were determined based on mycelial growth. The effective concentration 50 ranged from 0.11 to 15.92 µg/ml and 0.16 to 8.54 µg/ml, in 2014 and 2015, respectively. The frequency of low resistance to boscalid was 12.6 and 7.6%, and moderate resistance was 2.7 and 1.3% in 2014 and 2015, respectively. No highly resistant isolates were found in Henan Province, China. Mycelial growth, mycelial dry weight, spore production, and pathogenicity were not significantly different between resistant and sensitive phenotypes of the B. cinerea isolates. The results of cross-resistance testing showed no correlation between boscalid and carbendazim, procymidone, pyrimethanil, fluazinam, or fluopyram. In this study, the succinate dehydrogenase genes B (sdhB), C (sdhC), and D (sdhD) were analyzed and compared in sensitive and low-resistance and moderately resistant B. cinerea isolates to boscalid. Results showed that point mutations occurred simultaneously at sdhC amino acid positions 85 (G85A), 93 (I93V), 158 (M158V), and 168 (V168I) in 4 out of 10 sensitive isolates and 23 of 26 low-resistance and 5 of 5 moderately resistant B. cinerea isolates to boscalid. No point mutations were found in the sdhB and sdhD genes of all isolates. Furthermore, no point mutations were found in sdhB, sdhC, and sdhD genes in 3 of 26 low-resistance B. cinerea isolates to boscalid. Therefore, we speculate that the simultaneous point mutations in the sdhC gene may not be related to the resistance of B. cinerea to boscalid. These results suggested that there might be a substitution mechanism for the resistance of B. cinerea to the SDHI fungicide boscalid.


Occlusal tooth wear in male F344/N rats with aging.
With the aim of clarifying the aging properties of an animal model, the progress of occlusal tooth wear (OTW) of molars in male F344/N rats was monitored. Dried maxilla and mandible specimens from 61 male F344/N rats, aged 7 to >30 months, were used. The levels of OTW of all molars were monitored with aging. The cuspis dentis of molar teeth were worn out by 7 months (M) of age, and the occlusal surface became flat. As for each molar tooth (M(1-3), numbered in accordance with its position), OTW of M(1) was more severe in the lower than in the upper jaw, whereas M(3) was more severe in the upper than the lower jaw. OTW of M(2) in both the upper and the lower jaws progressed rapidly after 27M. OTW in male F344/N rats progressed faster than in females. However, when compensated for life span, both genders had similar profiles in OTW progress with aging. This study suggested that male rats were more convenient than females as a model for gerodontological research because of the earlier course of OTW progress.


Molecular cloning, overexpression and mapping of the slt gene encoding the soluble lytic transglycosylase of Escherichia coli.
The gene of the major autolysin of Escherichia coli, the soluble lytic transglycosylase (Slt), was isolated from an expression gene library. The cloned slt gene was used to determine its chromosomal map position adjacent to trp R at 99.7 min on the E. coli linkage map.


Rapid phase-modulated water excitation steady-state free precession for fat suppressed cine cardiovascular MR.
The purpose of this article is to describe a steady-state free precession (SSFP) sequence for fat suppressed cine cardiovascular magnetic resonance (CMR). A rapid phase-modulated binomial water excitation (WE) pulse is utilized to minimize repetition time and acquisition time. Three different water-excitation pulses were combined with cine-SSFP for evaluation. The frequency response of each sequence was simulated and examined in phantom imaging studies. The ratio of fat to water signal amplitude was measured in phantoms to evaluate the fat suppression capabilities of each method. Six volunteers underwent CMR of the heart at 1.5T to compare retrospectively-gated cine-SSFP with and without water excitation. The ratio of fat to myocardium signal amplitude was measured for conventional cine-SSFP and phase-modulated WE-SSFP. The proposed WE-SSFP method was tested in one patient referred for CMR to characterize a cardiac mass. The measured frequency response in a phantom corresponded to the numerical Bloch equation simulation demonstrating the widened stop-band around the fat resonant frequency for all water-excitation pulses tested. In vivo measurements demonstrated that a rapid, phase-modulated water excitation pulse significantly reduced the signal amplitude ratio of fat to myocardium from 6.92 +/- 2.9 to 0.8 +/- 0.13 (mean +/- SD) without inducing any perceptible artifacts in SSFP cine CMR. Fat suppression can be achieved in SSFP cine CMR while maintaining steady-state equilibrium using rapid, phase modulated, binomial water-excitation pulses.


Electron spin resonance study of human alpha 1-acid glycoprotein interaction with a spin labelled steroid.
The interaction of human alpha 1-acid glycoprotein (AAG) with a corticosteroid was studied using nitroxide labeled deoxycorticosterone and electron spin resonance (ESR) spectroscopy. The ESR spectra of the spin labeled steroid in the presence of AAG could be used to characterize the ligand-protein interaction at equilibrium without the need of a separation between bound and free species. An association constant Ka of 6.10(5) M-1 at 20 degrees C and a binding capacity of one site per mole protein were found. ESR spectra recorded at equilibrium at various temperatures allowed the calculation of enthalpy and entropy variations for the steroid-protein interaction; these thermodynamic parameters exhibited a rapid change above 45 degrees C which may be related to a protein conformational modification above this temperature, as detected by circular dichroism study. The ESR spectra width could be used to define a polar character for the spin label environment in the steroid binding site of AAG and to calculate an apparent rotational correlation time of 2.8 x 10(-8) sec for the steroid-protein complex in aqueous solution at 20 degrees C. It can be concluded that spin labeling and ESR methodology is of value in the study of steroid-protein interactions of biological significance above all because it can provide direct physico-chemical information concerning the local environment of the ligand in its binding site at equilibrium.


Sulfur-containing amino acids in 7TMRs: molecular gears for pharmacology and function.
Seven-transmembrane receptors (7TMRs) mediate the majority of physiological responses to hormones and neurotransmitters in higher organisms. Tertiary structure stability and activation of these versatile membrane proteins require formation or disruption of complex networks of well-recognized interactions (such as H-bonds, ionic, or aromatic-aromatic) but also of other type of interactions which have been less studied. In this review, we compile evidence from crystal structure, biophysical, and site-directed mutagenesis data that indicate or support the importance of interactions involving Met and Cys in 7TMRs in terms of pharmacology and function. We show examples of Met/Cys-aromatic and Met-Met interactions participating in ligand binding, in tuning the orientation of functionally important aromatic residues during activation or even in modulating the type of signaling response. Collectively, data presented enlarge the repertoire of interactions governing 7TMR functioning.


Lens reabsorption following self-induced needling and subsequent intracapsular secondary intraocular lens placement.
We present the case of a young man who inadvertently penetrated his cornea and lens with a needle used for injecting heroin. Three years later, the lens had completely reabsorbed leaving a fibrosed capsular bag. We describe the surgical techniques used to insert a secondary intraocular lens into the capsular bag with excellent visual outcome.


Association between high nasopharyngeal viral load and disease severity in children with human metapneumovirus infection.
Previous studies have shown that viral genotype and viral load may play a significant role in the pathogenesis of viral infections. The aim of this study was to evaluate these aspects of hMPV infections in children and their household contacts. Between 1 November 2003 and 31 March 2004, we prospectively studied 2060 children attending our Emergency Department for acute reasons. Nasopharyngeal swabs were collected upon enrolment and then tested with real-time PCR assays for the major viral causes of respiratory illness. Sixty children (2.9%) were infected by hMPV: 24 (1.2%) by hMPV A, 14 (0.7%) by hMPV B, 11 (0.5%) by untyped hMPV, and 11 (0.5%) by hMPV and an additional respiratory virus. There were no differences in disease presentation or in clinical or socioeconomic impact in relation to viral genotypes. HMPV viral load was significantly higher in children with lower respiratory tract involvement (p<0.05), hospitalised children (p<0.05), and the prevalence of secondary cases of a similar disease in the household of index cases (p<0.05). A high hMPV viral load correlated with disease presentation, whereas the overall clinical and socioeconomic burden caused by the two hMPV genotypes was similar.


Spontaneous inversion of left atrial appendage.
Inversion of the left atrial appendage is a rare phenomenon and frequently occurs as a complication following cardiac operations. Spontaneous inversion is even rarer, and so far, only three cases have been reported. We report an additional case of spontaneous inversion in 4-month female with Down's syndrome, hypothyroidism, and complete atrioventricular canal defect.


The ability of Hepatitis B surface antigen DNA vaccine to elicit cell-mediated immune responses, but not antibody responses, was affected by the deglysosylation of S antigen.
Hepatitis B Virus (HBV) infection remains a major worldwide infectious disease with serious long-term morbidity and mortality. The limited selections of drug treatment are not able to control the progress of disease in people with active and persistent HBV infection. Immunotherapy to control the degree of viral infection is one possible alternative solution to this challenge. HBV DNA vaccines, with their strong ability to induce cell-mediated immune responses, offer an attractive option. HBV surface protein is important in viral immunity. Re-establishing anti-S immunity in chronic HBV infected patients will bring significant benefit to the patients. Previous studies have shown that HBV S DNA vaccines are immunogenic in a number of animal studies. In the current study, we further investigated the effect of glycosylation to the expression and immunogenicity of S DNA vaccines. Our results demonstrate that deglycosylation at the two potential N-linked glycosylation sites in S protein resulted in a significant decrease of S-specific cell-mediated immune responses, but did not affect anti-S antibody responses. This finding provides important direction to the development of S DNA vaccines to elicit the optimal and balanced antibody and cell-mediated immune responses to treat people with HBV chronic infections.


Determining potential pregnancy status differences based on a new method of yearling heifer prebreeding examination.
The study objective was to evaluate the Ready-Intermediate-Problem (RIP) replacement heifer evaluation matrix's ability to classify heifers into groups with differing reproductive outcomes. Beef heifers (n = 341) from six Kansas herds were classified according to RIP matrix guidelines and then exposed to AI breeding, bull breeding, or a combination of both as per the management plans for each participating herd. Following the breeding season the heifers were evaluated to determine pregnancy status, AI pregnancy status, days bred, and the number of 21 day cycles needed during the breeding season to become pregnant. After the breeding season, 298 (87%) of the heifers were pregnant, 204 (68%) of which became pregnant in the first 21 days of the breeding season. There was a significant interaction (P = 0.01) in RIP classification and pregnancy by 21 day cycle. Ready classified heifers had a significantly greater risk of becoming pregnant after a single AI exposure (P = 0.03) and in the first 21-day cycle (P = 0.02) compared to Problem classified heifers, and significantly less risk of being non-pregnant at the end of the breeding season (P < 0.01) compared to Problem classified heifers. The RIP matrix can be useful for classifying heifers prior to the onset of the breeding season. Further research is needed to evaluate the matrix in other settings and populations of U.S. beef heifers as well as at different intervals between evaluation and the start of breeding season.


DNA microarray technologies for measuring protein-DNA interactions.
DNA-binding proteins have key roles in many cellular processes, including transcriptional regulation and replication. Microarray-based technologies permit the high-throughput identification of binding sites and enable the functional roles of these binding proteins to be elucidated. In particular, microarray readout either of chromatin immunoprecipitated DNA-bound proteins (ChIP-chip) or of DNA adenine methyltransferase fusion proteins (DamID) enables the identification of in vivo genomic target sites of proteins. A complementary approach to analyse the in vitro binding of proteins directly to double-stranded DNA microarrays (protein binding microarrays; PBMs), permits rapid characterization of their DNA binding site sequence specificities. Recent advances in DNA microarray synthesis technologies have facilitated the definition of DNA-binding sites at much higher resolution and coverage, and advances in these and emerging technologies will further increase the efficiencies of these exciting new approaches.


A case of cerebral aspergillosis caused by Aspergillus nidulans. Clinical, pathologic and mycologic identifications.
A case of cerebral aspergillosis is reported, the presenting symptom was numbness of right face, which worsened after one year. CT-scan showed two enhanced low-density patches in the anterior and basal parts of right temporal lobe. During operation, an abscess in the deep part of right temporal lobe was revealed. The patient gradually felt amaurosis and oculomotor palsy of right eye. About six months later, she died from intracranial hypertension. Biopsy, as well as autopsy findings suggested fungal infection, and was identified as Aspergillus nidulans, which has probably never been reported in the literature.


Comparing the Efficiency of Online and Field-Based Outreach for the Recruitment of Black and Latino Sexual Minority Men into an HIV Prevention Implementation Trial.
Rates of HIV diagnoses among young Black and Latino sexual minority men (SMM) have continued to increase since 2011; meanwhile, overall rates in the USA have decreased. Despite their importance, academic and medical institutions have often struggled to engage and recruit racial and ethnic minority SMM in HIV prevention services and research. The current study compares the success of two strategies for recruiting racial and ethnic minority SMM. Recruitment occurred in the context of a larger implementation study testing the effectiveness of a substance use and HIV prevention intervention among SMM at high risk for HIV infection. SMM (n = 778) were reached through either (1) field-based outreach conducted by two local community-based organizations (CBOs) delivering the intervention or (2) online recruitment coordinated by the trial's academic research partner. Field-based recruitment reached a significantly larger proportion of Black (42.9% vs. 18.2% reached online) and Latino individuals (40.3% vs. 28.1% reached online). Although online recruitment reached a greater proportion of SMM who met trial eligibility criteria (58.4% vs. 35.3% for field-based outreach; χ2(1) = 38.471, p < .001), a greater proportion of eligible participants identified through field-based outreach actually enrolled into the study trial (30.9% vs. 18.8% for online recruitment; χ2(1) = 7.82, p < .01). As a result, field-based recruitment required fewer recruitment staff hours (2.62 per enrolled participant) than online recruitment (3.46 recruitment staff hours per enrolled participant). Findings illustrate the potential for field-based, CBO-executed recruitment to reach Black and Latino SMM and for CBOs to successfully enroll those identified as eligible. Future prevention and implementation research should routinely engage community partners in the development of study recruitment strategies in a manner that can inform outreach and marketing efforts for HIV prevention research and prevention. Clinicaltrials.gov = NCT03488914.


Genetic influence on phenotypic differentiation in adult hippocampal neurogenesis.
Regulation of adult hippocampal neurogenesis has different regulatory levels, including cell proliferation, survival and differentiation. Cell proliferation and survival are differentially influenced by inheritable traits and the genetic background determines which regulatory levels of adult hippocampal neurogenesis are preferentially involved in a neurogenic response to environmental stimuli. We here compared baseline adult neurogenesis in wild-derived strain Mus spretus and three inbred laboratory strains: A/J, C3H/HeJ and DBA/2J. Proliferation of was similar in the four strains, with the extremes being A/J, which had about 2100+/-570 (mean+/-S.D.) labeled newborn cells per dentate gyrus (after 6 days of bromodeoxyuridine injections), and DBA/2J, which had approximately 1400+/-260. C3H/HeJ had approximately 1500+/-600 and M. spretus had 1550+/-270. Survival of new cells after 4 weeks was 19% in A/J and DBA/2J, and 21% in M. spretus, but 37% in C3H/HeJ. Survival in C3H/HeJ was significantly different from DBA/2. Phenotypic analysis revealed that DBA/2J produced significantly fewer new neurons than A/J and C3H/HeJ (47% vs. 63% and 67%) but significantly more new astrocytes than A/J and C3H/HeJ (28% vs. 9% and 11%). In absolute terms there were 370+/-120 new neurons in C3H/HeJ, 250+/-60 in A/J, 130+/-50 in DBA/2J, and 190+/-130 in M. spretus. Our results indicate that regulation of adult hippocampal neurogenesis affects the level of phenotypic differentiation. At the present time it cannot be determined whether this regulation occurs by influencing cell fate decisions or by promoting selective survival.


Association between Goals of Care Designation orders and health care resource use among seriously ill older adults in acute care: a multicentre prospective cohort study.
The Goals of Care Designation (GCD) is a medical order used to communicate the focus of a patient's care in Alberta, Canada. In this study, we aimed to determine the association between GCD type (resuscitative, medical or comfort) and resource use during hospitalization. This was a prospective cohort study of newly hospitalized inpatients in Alberta conducted from January to September 2017. Participants were aged 55 years or older with chronic obstructive pulmonary disease, congestive heart failure, cirrhosis, cancer or renal failure; aged 55-79 years and their provider answered "no" to the "surprise question" (i.e., provider would not be surprised if the patient died in the next 6 months); or aged 80 years or older with any acute condition. The exposure of interest was GCD. The primary outcome was health care resource use during admission, measured by length of stay (LOS), intensive care unit hours, Resource Intensity Weights (RIWs), flagged interventions and palliative care referral. The secondary outcome was 30-day readmission. Adjusted regression analyses were performed (adjusted for age, sex, race and ethnicity, Clinical Frailty Scale score, comorbidities and city). We included 475 study participants. The median age was 83 (interquartile range 77-87) years, and 93.7% had a GCD at enrolment. Relative to patients with the resuscitative GCD type, patients with the medical GCD type had a longer LOS (1.42 times, 95% confidence interval [CI] 1.10-1.83) and a higher RIW (adjusted ratio 1.14, 95% CI 1.02-1.28). Patients with the comfort and medical GCD types had more palliative care referral (comfort GCD adjusted relative risk (RR) 9.32, 95% CI 4.32-20.08; medical GCD adjusted RR 3.58, 95% CI 1.75-7.33) but not flagged intervention use (comfort GCD adjusted RR 1.06, 95% CI 0.49-2.28; medical GCD adjusted RR 0.98, 95% CI 0.48-2.02) or 30-day readmission (comfort GCD adjusted RR 1.00, 95% CI 0.85-1.19; medical GCD adjusted RR 1.05, 95% CI 0.97-1.20). Goals of Care Designation type early during admission was associated with LOS, RIW and palliative care referral. This suggests an alignment between health resource use and the focus of care communicated by each GCD.


The value of hyaluronidase treatment of different tissues before demonstration of fibronectin by the indirect immunoperoxidase technique.
The influence of testicular hyaluronidase treatment on the immunohistochemical localization of fibronectin in different tissues (human articular cartilage, large intestine, synovial membrane and experimental granulation tissue) as well on frozen as on formaldehyde fixed, paraffin embedded tissue, has been studied using the indirect immunoperoxidase technique. Pretreatment with hyaluronidase is essential in demonstrating fibronectin in frozen sections of human articular cartilage. In the other tissues examined treatment with hyaluronidase was not essential, but gave a more optimal staining quality. The effect of hyaluronidase treatment was to some extent dependent on the duration of treatment. In formaldehyde fixed, paraffin processed tissue the improvement with hyaluronidase treatment was only seen when the hyaluronidase followed pepsin digestion of the deparaffinized tissue sections.


Stress and Stressors: The Impact of the COVID-19 Pandemic on Students, Faculty and Staff at a Historically Black College/University.
Due to COVID-19 restrictions, academic institutions have changed their modus operandi, particularly in adopting distance learning in lieu of face-to-face instruction. This has sometimes produced unanticipated effects on students. The purpose of this study was to determine COVID-19 pandemic stressors and coping mechanisms utilized as relief measures by students, faculty, and staff in the College of Health Sciences at a historically Black institution. Cross-sectional study. A convenience sample of 209 students, 34 faculty, and 9 staff from the College of Health Sciences at a historically Black institution participated in this study. A 32-item Qualtrics survey was utilized to gather demographic data, COVID-19-related stressors, and coping mechanisms. Descriptive statistics, Chi-square test for categorical variables, and Cochran-Mantel-Haenszel tests for ordinal variables were used. Female students, African American students (AAS), and undergraduates perceived stress related to the risk of contagion during the pandemic higher than the male students (P-value = 0.0096), other races (P-value = 0.0249) and graduate students (P-value = 0.0141) respectively. Female students perceived more stress related to relationships with relatives (P-value = 0.0128). Caucasian students compared to others (P-value = 0.0240) and graduate students compared to undergraduate students (P-value = 0.0011) reported less perceived stress related to the relationships with other colleagues. Top coping strategies of students and staff included interaction with family and friends, social media, hand hygiene, and meditation/spiritualty. Except for "avoidance of public spaces/transportation", all other mechanisms were the same for faculty.


Hypertriglyceridemia in an outpatient department--Significance as an atherosclerotic risk factor.
Although a relationship between elevated triglycerides (TG) and cardiovascular diseases is generally accepted, its extent is still discussed. This retrospective study analyzed the incidence of cardiovascular events (CVE) and pancreatitis as well as the therapeutic regimen in patients being treated for hypertriglyceridemia (HTG) at an outpatient department. The cohort included 183 patients with mild and 49 patients with severe HTG; subgroups were formed and compared according to gender, presence of metabolic vascular syndrome (MVS) and lipid values. Patients in this study seem to have had CVE at younger age than reported event rates in the general population. TG levels, rates of CVE and pancreatitis were reduced in all groups during therapy, which could be linked to use of omega-3 fatty acids and fibrates. Patients with persisting severe HTG as a result of incompliance showed massive risk for pancreatitis. Although no significant association between TG levels and CVE could be established, the combination of HTG and other cardiovascular risk factors such as MVS seems to be especially dangerous. The lipid-lowering drug therapy appeared to be effective with respect to CVE and pancreatitis incidence.


Ceramides and Acute Kidney Injury.
Altered lipid metabolism is a characteristic feature and potential driving factor of acute kidney injury (AKI). Of the lipids that accumulate in injured renal tissues, ceramides are potent regulators of metabolism and cell fate. Up-regulation of ceramide synthesis is a common feature shared across several AKI etiologies in vitro and in vivo. Furthermore, ceramide accumulation is an early event in the natural history of AKI that precedes cell death and organ dysfunction. Emerging evidence suggests that inhibition of ceramide accumulation may improve renal outcomes in several models of AKI. This review examines the landscape of ceramide metabolism and regulation in the healthy and injured kidney. Furthermore, we discuss the body of literature regarding ceramides as therapeutic targets for AKI and consider potential mechanisms by which ceramides drive kidney pathogenesis.


Subjective wellbeing and emotion regulation strategies: How are they associated with student engagement in online learning during Covid-19?
The COVID-19 pandemic has brought unprecedented challenges to the world, creating significant impact on people's lives and subjective wellbeing. While previous studies have shown that students' wellbeing and how they manage their emotions are critical to students' learning, less research has considered their specific impacts on student engagement in online learning. The aim of this study is to examine how students' subjective wellbeing and emotion regulation strategies (viz cognitive reappraisal and expressive suppression) are associated with student engagement in online learning during the pandemic. A total of 965 students from a university in China participated in the study. The data were collected online during the COVID-19 from March to July 2020, which included measures of wellbeing, emotion regulation strategie, and online learning engagement. Structural equation modelling results showed that wellbeing was positively associated with both the use of reappraisal and suppression. Moreover, mediation analysis showed that reappraisal partially mediated the relationship between wellbeing and all types of online learning engagement (including cognitive, emotional and behavioural engagements). Conversely, suppression was found to have a small mediating effect between wellbeing and behavioural engagement of online learning only. No such effect was found on cognitive and emotional engagements. The findings provide evidence on the positive relationship between wellbeing and both the strategies of suppression and reappraisal during the pandemic. However, only reappraisal has positive impact on learning, suggesting the importance of effective regulation strategies on students' online engagement. Theoretical and practical implications of the findings are discussed.


[Pathologic femoral neck fractures and para-articular beta 2 microglobulin amyloidosis in long-term dialysis patients: an increasingly frequent disease picture].
Beta-2-microglobulin (beta 2-M)-amyloidosis is caused by retention of beta 2-M by dialyzing membranes and by deposition as an atypical amyloid in synovia, bone and tendons. Ten patients under long-term hemodialysis (13 +/- 2.7 years) were treated because of hip pain and cystiform skeletal alterations. First typical joint affections appeared nine years after start of hemodialysis. Five fractures of the hip neck due to large cystiform bone lesions occurred in four subjects. In all cases a total joint replacement was performed.


An improved method for detecting circulating microRNAs with S-Poly(T) Plus real-time PCR.
We herein describe a simple, sensitive and specific method for analysis of circulating microRNAs (miRNA), termed S-Poly(T) Plus real-time PCR assay. This new method is based on our previously developed S-Poly(T) method, in which a unique S-Poly(T) primer is used during reverse-transcription to increase sensitivity and specificity. Further increased sensitivity and simplicity of S-Poly(T) Plus, in comparison with the S-Poly(T) method, were achieved by a single-step, multiple-stage reaction, where RNAs were polyadenylated and reverse-transcribed at the same time. The sensitivity of circulating miRNA detection was further improved by a modified method of total RNA isolation from serum/plasma, S/P miRsol, in which glycogen was used to increase the RNA yield. We validated our methods by quantifying miRNA expression profiles in the sera of the patients with pulmonary arterial hypertension associated with congenital heart disease. In conclusion, we developed a simple, sensitive, and specific method for detecting circulating miRNAs that allows the measurement of 266 miRNAs from 100 μl of serum or plasma. This method presents a promising tool for basic miRNA research and clinical diagnosis of human diseases based on miRNA biomarkers.


Experiences of nurse practitioners working during the COVID-19 pandemic: A metasynthesis of qualitative studies.
The impact of the COVID-19 pandemic forced global changes to how nurses practice. Nurse practitioners adjusted their scope, changed how they delivered their service, and worked with limited resources. For some services, patient access was also compromised. To synthesize, combine, and present current evidence on the experiences of nurse practitioners working during the COVID-19 pandemic. CINAHL, Embase, and MEDLINE electronic databases were used to perform a structured search strategy. During the COVID-19 pandemic, health care services had to leverage their workforce skills to accelerate COVID-19 identification, treatment, and care. Nurse practitioners rapidly found themselves at the forefront and were concerned about infecting others. They also identified the need for support and were able to adapt to the changing environment. Nurse practitioners also recognized the impact on their well-being. Having insight into nurse practitioner's experiences during the pandemic is valuable for future health care workforce planning. Understanding how they coped will help with critical preparedness and response actions to other health care crises. Having insight into nurse practitioner's experiences during the pandemic is valuable for future health care workforce planning because, as we know, the nurse practitioner workforce is one of the most rapidly growing professions in primary health care. Any future work in this space will help inform future nurse practitioner education and also help by informing critical preparedness and response actions to future health care crises, whether global or local or clinical or nonclinical.


Central and Local Modulators of Reproduction and Fertility: An Update.
Infertility is currently one of the most important health troubles in industrialised countries after cardio-vascular diseases and cancer [...].


Neighbourhood relationships moderate the positive association between family caregiver burden and psychological distress in Japanese adults: a cross-sectional study.
Family caregiver burden is associated with higher psychological distress. However, little is known about the impact of neighbourhood relationships on caregivers' psychological distress. We examined whether neighbourhood relationships of caregivers moderate the association between family caregiver burden and psychological distress. This was a cross-sectional study. We recruited 5321 Japanese adults who participated in the Japan Multi-Institutional Collaborative Cohort Study in the Okazaki area between 2013 and 2017. Participants completed self-reported questionnaires to measure psychological distress (Kessler 6: K6), subjective caregiver burden, and neighbourhood relationships. We performed a multivariable linear regression analysis in which caregiver burden was designated as an independent variable and the K6 score as a dependent variable, adjusting for demographics. The interaction term between caregiver burden and neighbourhood relationships was also included in the analysis. Data from a total of 5069 participants were included (mean age [standard deviation]: 63.1 years [10.3 years]; 2226 [43.9%] female). Caregiver burden was significantly and positively associated with psychological distress (compared with no burden, mild burden: β = 0.24, P = 0.197; severe burden: β = 0.60, P < 0.01; P for trend < 0.01). There was a significant negative interaction effect of caregiver burden × neighbourhood relationship on psychological distress (severe burden × good neighbourhood relationship: β = -3.29, P < 0.01). A higher caregiver burden was associated with higher psychological distress, and neighbourhood relationships moderated this association. Our findings suggest that good neighbourhood relationships can buffer caregiving-associated psychological distress.


A Supported Nickel Catalyst Stabilized by a Surface Digging Effect for Efficient Methane Oxidation.
A surface digging effect of supported Ni NPs on an amorphous N-doped carbon is described, during which the surface-loaded Ni NPs would etch and sink into the underneath carbon support to prevent sintering. This process is driven by the strong coordination interaction between the surface Ni atoms and N-rich defects. In the aim of activation of C-H bonds for methane oxidation, those sinking Ni NPs could be further transformed into thermodynamically stable and active metal-defect sites within the as-generated surface holes by simply elevating the temperature. In situ transmission electron microscopy images reveal the sunk Ni NPs dig themselves adaptive surface holes, which would largely prevent the migration of Ni NPs without weakening their accessibility. The reported two-step strategy opens up a new route to manufacture sintering-resistant supported metal catalysts without degrading their catalytic efficiency.


Insulin and isoproterenol induce phosphorylation of the particulate cyclic GMP-inhibited, low Km cyclic AMP phosphodiesterase (cGI PDE) in 3T3-L1 adipocytes.
The cGI PDE in particulate fractions of differentiated adipocytes (but not control 3T3-L1 fibroblasts) was cross-reactive with a polyclonal antibody raised against the bovine adipose cGI PDE. The 3T3-L1 adipocyte cGI PDE is a 135 kDa protein which is phosphorylated in 32P-labeled cells in response to beta-agonist or insulin. These results indicate that the 3T3-L1 cGI PDE is similar in structure and hormonal regulation to the analogous enzyme in the rat adipocyte.


A Phase II randomized controlled trial for lung and diaphragm protective ventilation (Real-time Effort Driven VENTilator management).
Lung Protective Mechanical Ventilation (MV) of critically ill adults and children is lifesaving but it may decrease diaphragm contraction and promote Ventilator Induced Diaphragm Dysfunction (VIDD). An ideal MV strategy would balance lung and diaphragm protection. Building off a Phase I pilot study, we are conducting a Phase II controlled clinical trial that seeks to understand the evolution of VIDD in critically ill children and test whether a novel computer-based approach (Real-time Effort Driven ventilator management (REDvent)) can balance lung and diaphragm protective ventilation to reduce time on MV. REDvent systematically adjusts PEEP, FiO2, inspiratory pressure, tidal volume and rate, and uses real-time measures from esophageal manometry to target normal levels of patient effort of breathing. This trial targets 276 children with pulmonary parenchymal disease. Patients are randomized to REDvent vs. usual care for the acute phase of MV (intubation to first Spontaneous Breathing Trial (SBT)). Patients in either group who fail their first SBT will be randomized to REDvent vs usual care for weaning phase management (interval from first SBT to passing SBT). The primary clinical outcome is length of weaning, with several mechanistic outcomes. Upon completion, this study will provide important information on the pathogenesis and timing of VIDD during MV in children and whether this computerized protocol targeting lung and diaphragm protection can lead to improvement in intermediate clinical outcomes. This will form the basis for a larger, Phase III multi-center study, powered for key clinical outcomes such as 28-day ventilator free days. Clinical Trials Registration: NCT03266016.


Seroprevalence and risk assessment of Toxoplasma gondii infection in sheep and goats in North and Beqaa governorates of Lebanon.
Toxoplasmosis is a disease caused by the protozoan parasite Toxoplasma gondii that affects both humans and animals, leading to abortions and significant clinical manifestations in pregnant and immunocompromised hosts, in addition to massive economic losses in animal industries. Data from Lebanon are scarce regarding the seroprevalence of T. gondii infection in livestock. This study aimed to estimate the seroprevalence and assess the associated risk factors of T. gondii infection in sheep and goats in Lebanon. A cross-sectional study was carried out from May 2020 to April 2021. Blood samples from 150 sheep and 145 goats (total 295) destined for human consumption were obtained from 20 Lebanese farms located in the North and Beqaa governorates. The anti-T. gondii immunoglobulin G antibodies were assayed through means of a modified agglutination test with a cutoff titer of 20. An overall seroprevalence of 48.5% (143/295) was reported: About 56.6% seroprevalence was found in sheep (85/150) and 40% (58/145) in goats. Adult age, female gender, and the wet season were significantly associated with an increased seropositivity rate of T. gondii infection (p < 0.001, p = 0.001, and p = 0.043, respectively). These results confirm the spread of T. gondii in sheep and goats destined for human consumption in various geographical regions in Lebanon. Therefore, continuous monitoring of T. gondii infection in livestock is warranted to control the spread of the infection and limit its potential transmission to humans through the consumption of raw or undercooked meat.


Enterohemorrhagic Escherichia coli pathogenesis: role of Long polar fimbriae in Peyer's patches interactions.
Enterohemorrhagic Escherichia coli (EHEC) are major food-borne pathogens whose survival and virulence in the human digestive tract remain unclear owing to paucity of relevant models. EHEC interact with the follicle-associated epithelium of Peyer's patches of the distal ileum and translocate across the intestinal epithelium via M-cells, but the underlying molecular mechanisms are still unknown. Here, we investigated the involvement of Long polar fimbriae (Lpf) in EHEC pathogenesis. Of the 236 strains tested, a significant association was observed between the presence of lpf operons and pathogenicity. In sophisticated in vitro models of the human gastro-intestinal tract, lpf expression was induced during transit through the simulated stomach and small intestine, but not in the colonic compartment. To investigate the involvement of Lpf in EHEC pathogenesis, lpf isogenic mutants and their relative trans-complemented strains were generated. Translocation across M-cells, interactions with murine ileal biopsies containing Peyer's patches and the number of hemorrhagic lesions were significantly reduced with the lpf mutants compared to the wild-type strain. Complementation of lpf mutants fully restored the wild-type phenotypes. Our results indicate that (i) EHEC might colonize the terminal ileum at the early stages of infection, (ii) Lpf are an important player in the interactions with Peyer's patches and M-cells, and could contribute to intestinal colonization.


A methodological perspective on learning in the developing brain.
The brain undergoes profound development across childhood and adolescence, including continuous changes in brain morphology, connectivity, and functioning that are, in part, dependent on one's experiences. These neurobiological changes are accompanied by significant changes in children's and adolescents' cognitive learning. By drawing from studies in the domains of reading, reinforcement learning, and learning difficulties, we present a brief overview of methodological approaches and research designs that bridge brain- and behavioral research on learning. We argue that ultimately these methods and designs may help to unravel questions such as why learning interventions work, what learning computations change across development, and how learning difficulties are distinct between individuals.


Clinical outcomes after repair of left atrial esophageal fistulas occurring after atrial fibrillation ablation procedures.
The initial experience with left atrial esophageal fistula (LAEF) secondary to atrial fibrillation (AF) ablation procedures revealed a near-universal mortality. A comprehensive description of the principles of LAEF repair in the modern era and its resulting impact on morbidity and mortality are lacking in the literature. To describe the presentation, surgical management, and outcomes of patients with LAEF. A retrospective cohort analysis of 29 patients was performed, including previously unpublished cases of surgically repaired LAEF from 4 institutions (n = 6), and all published cases of surgically repaired (n = 16) or stented (n = 7) LAEF. The mean age was 55 ± 13 years, and 75% were men who were undergoing radiofrequency energy catheter ablation (n = 26), cryoablation (n = 1), high-intensity focused ultrasound ablation (n = 1), and surgical mini-MAZE procedure (n = 1) and presented 30 ± 12 days postablation procedure. Overall, 55% of the patients receiving an intervention for LAEF died (41% surgical repair; 100% stent). Patients who did not receive primary esophageal repair were more likely to experience postoperative complications, including mediastinitis, need for percutaneous endoscopic gastrostomy (PEG) feeds, esophageal stent, or death (P = .05). In addition, interposing tissue between the repaired esophagus and the left atrium resulted in fewer postoperative complications (P = .02). While improved relative to initial reports, mortality associated with LAEF remains high after corrective intervention. Primary esophageal repair with the placement of tissue between the repaired esophagus and the left atrium may result in lower morbidity and mortality.


The interface between chronic fatigue syndrome and depression: A psychobiological and neurophysiological conundrum.
The chronic fatigue syndrome (CFS) remains a contentious and controversial presentation despite decades of systematic research from a variety of medical specialties and associated disciplines. Variously championed as a condition of immunological, neurological, neurophysiological, psychiatric or psychological origin, consensus on a cogent and evidenced-based pathway has yet to be achieved. Irrespective of the ambiguity regarding aetiology, what is incontrovertible is the experience of significant depression, which often accompanies this most distressing clinical presentation. The current paper examines the potential underlying mechanisms, which may determine and explain this relationship between CFS and depression. In doing so, it offers some insights, which may be of value in the development of evidence-based and scientifically-anchored interventions in individuals experiencing this diagnosis, to improve outcomes in relation to depression specifically and quality of life more generally.


An unusual lethal complication of preservation of chordae tendineae in mitral valve replacement.
In two patients, several chordae tendineae of the mural leaflet were preserved during mitral valve replacement. Hemorrhagic necrosis and spontaneous rupture of the preserved posterior papillary muscle led to disc entrapment and the death of both patients.


Studies on drug metabolism by fungi colonizing decomposing human cadavers. Part I: DNA sequence-based identification of fungi isolated from postmortem material.
Cadavers can be colonized by a wide variety of bacteria and fungi. Some of these microbes could change the concentration or the metabolic pattern of drugs present in postmortem samples. The purpose of this study was to identify fungi from human postmortem material and to further assess their potential role in the metabolism of drugs. Aliquots of 252 postmortem samples (heart blood, liver, kidney, and lung) taken from 105 moderately to severely decomposed bodies were streaked on Sabouraud agar for isolation of fungal species. One part of the samples was worked up immediately after autopsy (group I). The second part had previously been stored at -20 °C for at least 1 year (group II). Identification of the isolates was achieved morphologically by microscopy and molecularly by polymerase chain reaction amplification and sequencing of markers allowing species identification of the respective genera. Depending on the genus, different gene fragments were used: calmodulin for Aspergillus, β-tubulin for Penicillium, translation elongation factor 1α for Fusarium, and the internal transcribed spacer region of the ribosomal DNA for all remaining genera. A total of 156 fungal strains were isolated from 62% of the postmortem materials. By using these primers, 98% of the isolates could be identified to the species level. The most common genera were Candida (60.0%-six species), Penicillium (10.3%-two species), Rhodotorula (7.1%-one species), Mucor (6.4%-four species), Aspergillus (3.2%-four species), Trichosporon (3.2%-one species), and Geotrichum (3.2%-one species). Group I samples contained 53% more fungal species than stored samples suggesting some fungi did not survive the freezing process. The isolated fungi might be characteristic for decomposed bodies. The proposed methodology proved to be appropriate for the identification of fungi in this type of material.


Development of a novel digital breath-activated inhaler: Initial particle size characterization and clinical testing.
Delivery of inhaled respiratory medications have been associated with variable delivery of drug due to errors in device operations and have not been designed to monitor true delivery of medication. A fully digital breath-activated inhaled (DBAI) delivery platform has been developed with integrated firmware and software to address these limitations. the device was designed to produce similar aerosol particle output to a marketed albuterol MDI and to the albuterol/ipratropium combination in a soft mist inhaler (SMI). Cascade impactor studies were conducted to demonstrate comparable aerodynamic particle size distribution (APSD) metrics. Efficacy was evaluated by pharmacodynamic studies involving spirometry in two separate protocols with adult subjects having COPD (albuterol DBAI vs. albuterol MDI - Study A, albuterol/ipratropium DBAI single arm - Study B). The total emitted doses (TED) were 81.9 ± 10.3, 109.3 ± 15.0 and 121.9 ± 7.0 μg/actuation for the DBAI, SMI and MDI respectively, and the fine (respirable) particle doses (FPD) were 56.2 ± 6.0, 61.7 ± 5.5 and 79.4 ± 2.7 μg/actuation. MMADs for albuterol sulfate were 1.93 ± 0.11, 1.75 ± 0.19, and 2.65 ± 0.05 μm for the DBAI, Respimat soft mist inhaler (SMI) and MDI respectively. The corresponding GSDs were 1.96 ± 0.16, 2.79 ± 0.25, and 1.48 ± 0.02 μm. The corresponding respirable fractions were 68.7 ± 3.2%, 57.3 ± 10.5%, and 65.2 ± 2.4%. Spirometric study A enrolled 23 subjects (age 64 ± 7.3 years, 39% male, FEV1 45 ± 14% predicted). Study B enrolled 23 subjects (age 65 ± 8.6 years, 43% male, FEV1 47 ± 10% predicted). For Study A, FEV1 at 20 min post-dose improved by 120 (167) mL (p = 0.002) for the DBAI device and 109 (183) mL (p = 0.008) for the MDI device (p = 0.86 for between group differences). For Study B, FEV1 (20 min post-dose) improved by 216 (126) mL (p < 0.001). The DBAI generated highly respirable aerosols containing albuterol sulfate that were similar to the MDI and SMI in respirable fraction but lower in dose. Subsequent pharmacodynamic studies delivering albuterol sulfate alone and in combination with ipratropium bromide confirmed similar responses for the DBAI compared with the other inhalers, which could possibly be related to a response ceiling. The DBAI breath-activated capability combined with the ability to monitor actual delivery of medication may improve effectiveness by overcoming patient miscoordination.


Antiarrhythmic effects of nicorandil on canine cardiac Purkinje fibers.
Antiarrhythmic efficacy of nicorandil (SG-75) (1-100 microM), a coronary vasodilator, was investigated electrophysiologically with regard to the action potentials of canine Purkinje fibers. The main results obtained are as follows: (a) nicorandil suppressed three kinds of automaticities, i.e., spontaneous or low-K+-induced automaticity and electrical depolarization-induced automaticity; (b) the drug increased the effective refractory period relative to action potential duration and decreased differences between action potential duration and effective refractory period; and (c) in the presence of nicorandil (50 microM), membrane potentials at which the earliest premature response could be elicited were significantly more negative than control, thus leading to a much faster upstroke velocity of the premature response. These changes in electrophysiological properties suggest that the drug may be effective for treating selected cardiac arrhythmias due to both enhanced automaticity and reentry, particularly in the presence of a decreased membrane K conductance. These effects of nicorandil could be mostly attributed to an increase in membrane K conductance.


Gene amplification and overexpression of HER2 in renal cell carcinoma.
To determine the frequency of HER2 genetic abnormalities in renal cell carcinoma (RCC) and hence assess the potential suitability of Herceptin immunotherapy. Tumours from 27 patients with RCC were assessed; all patients had undergone nephrectomy. Benign renal tissue from the nephrectomy specimens was studied in seven patients. Gene amplification was assessed using fluorescent in-situ hybridization, and protein over-expression using immunohistochemistry. Twenty-four patients had clear cell renal carcinoma, two had papillary renal carcinoma and one a sarcomatoid variant carcinoma. There was no HER2 amplification in the tumours or the benign renal tissue. Polysomy 17 was detected in 11 of 27 tumours (41%) and increased HER2 copy number in seven (26%). Both polysomy 17 and increased HER2 copy number were absent in the benign renal tissue. Three tumours (11%) and six of the seven benign renal tissue samples over-expressed the HER2 protein. HER2 amplification is absent and protein over-expression uncommon in RCC. This casts doubt on the suitability of Herceptin in the treatment of RCC.


Relationship between blood viscosity and cerebral ischemia after surgical treatment of ruptured intracranial aneurysms.
To determine the role of blood viscosity after surgical treatment of ruptured intracranial aneurysms, the relationship between blood viscosity and clinical condition was examined in 17 patients. A total of 213 blood samples were analyzed. An inverse correlation was found between blood viscosity and level of consciousness; in addition, blood viscosity was higher when focal neurologic deficit was observed. Hematocrit was similarly related to clinical condition, although the correlations observed were less strong. Postoperative plasma viscosity was higher in patients with focal neurologic deficit. Regular blood viscosity measurements are of value in patients at risk for developing cerebral ischemia.


Antimalarial activity of phenazines from lapachol, beta-lapachone and its derivatives against Plasmodium falciparum in vitro and Plasmodium berghei in vivo.
The antimalarial activity of benzo[a]phenazines synthesized from 1,2-naphthoquinone, lapachol, beta-lapachone and several derivatives have been tested against Plasmodium falciparum in vitro using isolates of parasites with various susceptibilities to chloroquine and/or mefloquine. Parasite growth in the presence of the test drugs was measured by incorporation of [(3)H]-hipoxanthine in comparison to controls with no drugs, always testing in parallel chloroquine, a standard antimalarial. Among seven benzophenazines tested, four had significant in vitro activities; important, the parasites resistant to chloroquine were more susceptible to the active phenazines in vitro. The doses of phenazines causing 50% inhibition of parasite growth varied from 1.67 to 9.44 microM. The two most active ones were also tested in vivo against Plasmodium berghei in mice, in parallel with lapachol and beta-lapachone. The 3-sulfonic acid-beta-lapachone-derived phenazine was the most active causing up to 98% inhibition of parasitaemia in long term treatment (7 doses) subcutaneously, whereas the phenazine from 3-bromo-beta-lapachone was inactive. Thus, these simple phenazines, containing polar (-Br,-I) and ionizable (-SO(3)H, -OH) groups, easily synthesized from cheap, natural or synthetic precursors (lapachol and beta-lapachone), at rather low cost, provide prototypes for development of new antimalarials aiming the chloroquine resistant parasites.