Innervation zone targeted botulinum toxin injections.
Muscle overactivity (spasticity, dystonia or spasm) seen in certain neuromuscular disorders has been effectively treated with intramuscular injection of botulinum neurotoxins (BTXs). Since they act in the nerve terminals, toxins must be transported to the neuromuscular junctions which are generally clustered in one or more restricted areas (innervations zone(s) (IZs)) in a skeletal muscle. Likewise, IZ targeted BTX injections using guidance is highly recommended to achieve an optimal therapeutic goal with lower doses and fewer side effects. Hence, detection of the injection sites should be based on the knowledge about the localization of the IZs and also the transport mechanism of BTX in skeletal muscle is crucial for intramuscolar application of BTX. In this paper, IZ(s) of the skeletal muscles, distribution of BTX, proper sites and guidance for the injections are discussed taking into account the muscle structure and architecture.


Cholinergic basis of memory-strengthening effect of Foeniculum vulgare Linn.
Alzheimer's disease is a neurodegenerative disorder associated with a decline in cognitive abilities. Dementia is one of the age-related mental problems and a characteristic symptom of Alzheimer's disease. Nootropic agents are used in situations where there is organic disorder in learning abilities. The present work was undertaken to assess the potential of Foeniculum vulgare Linn. extract as a nootropic and anticholinesterase agent in mice. Methanolic extract of the whole plant of F. vulgare Linn. administered for eight successive days ameliorated the amnesic effect of scopolamine (0.4 mg/kg) and aging- induced memory deficits in mice. The passive avoidance paradigm served as the exteroceptive behavioral model for assessing memory. F. vulgare extract increased step-down latency and acetylcholinesterase inhibition in mice significantly. Hence, F. vulgare can be employed in treatment of cognitive disorders such as dementia and Alzheimer's disease.


Comprehensive expression profile of long non-coding RNAs in Peripheral blood mononuclear cells from patients with neuropsychiatric systemic lupus erythematosus.
Long noncoding RNAs (lncRNAs) are emerging as critical regulators of gene expression in the immune system, but their impact on neuropsychiatric systemic lupus erythematosus (NPSLE) remains unknown. RNA sequencing analysis was used to screen the comprehensive expression profile of lncRNAs and messenger RNAs (mRNAs) in peripheral blood mononuclear cells (PBMCs) from NPSLE patients, active SLE patients who had never experienced neuropsychiatric manifestations (Non-NPSLE) and healthy controls. Differentially expressed (DE) lncRNA levels were validated by qRT-PCR in 26 NPSLE patients, 31 Non-NPSLE patients and 30 healthy controls. Further, correlations of DE lncRNAs with clinical manifestations of NPSLE patients were analyzed. Finally, a bioinformatic analysis was performed to investigate the potential functions of DE genes. Four hundred and fifty-one lncRNAs and 272 mRNAs were DE between the NPSLE patients and Non-NPSLE patients, among which, significantly upregulated expression levels of NONHSAT208182.1, NONHSAT182114.1, NONHSAT106801.2, NONHSAT039491.2, ENST00000356215, NONHSAT087499.2 and NONHSAT207026.1 while downregulated expression levels of NONHSAT001281.2 and NONHSAT024353.2 were further validated in PBMCs from NPSLE patients by qRT-PCR. Bioinformatic analysis suggested several gene ontology (GO) terms and signal pathways may play important roles in NPSLE development. Co-expression networks analysis indicated that 170 lncRNAs and 46 mRNAs were included in the co-expression network. The expression level of NONHSAT039491.2 was associated with the activity of SLE and the presence of anti-dsDNA, anti-RNP antibody, dizziness and headache. NONHSAT087499.2 level correlated with anti-RNA antibody, ENST00000356215 level correlated with olfactory threshold and oral ulcer. NONHSAT208182.1 level correlated with the presence of fever, unstable walking and urinary red blood cells. NONHSAT106801.2 correlated with frequency of B cells and the presence of fever. NONHSAT024353.2 level was associated with serum IgG levels and the presence of anti-SSA and disorder of consciousness. Our data provided comprehensive evidence regarding the differential expression of lncRNAs in PBMCs from NPSLE patients, indicating that these DE lncRNAs may play roles in NPSLE development. Our finding shed light on the understanding of the molecular mechanisms of lncRNAs in the pathogenesis of NPSLE.


History of instrumentation for stabilization of the subaxial cervical spine.
In the past several decades methods have been developed to stabilize the subaxial cervical spine both posteriorly and anteriorly. Methods of posterior stabilization have progressed from interspinous wiring, through facet wiring and sublaminar wiring, to the lateral mass screws with plates and rods that are in use today. Plates for anterior stabilization have evolved from rigid plates requiring bicortical screws through those used with unicortical locking screws, to dynamic load-sharing plates used with variable angle screws. The original description of spinous process wiring was published by Hadra in 1891. In 1942 Rogers described the interspinous wiring method used for trauma-induced cervical instability, which was modified by Bohlman in 1985 (triple wiring technique). Luque rods with sublaminar wires were introduced in the late 1970s to address multilevel and occipitocervical instability. Facet wiring was developed in 1977 by Callahan to address the problem of stabilization when laminae are not present. Wiring remained the method used until Roy-Camille introduced the lateral mass screw-plate construct in the 1980s. The first plate for anterior stabilization was designed by Orozco and Llovet in 1970 and was later refined by Caspar; this was a rigid plate with bicortical screws. Morscher devised unicortical locking screws in the 1980s. The latest concept of dynamic load-sharing plates with variable angle screws was developed in 2000. In this article historical landmarks in surgical methods for the stabilization of the subaxial cervical spine are reviewed.


[From the standpoint in department of geriatrics in university hospital].
Because Japanese society has increasing elderly persons and also has a low birth rate. Medical treatments and benefits will be severely limited in the future because of economic problems. I would like to look back on the history of Japanese Geriatrics Society and the many studies on geriatrics in Japan. The first annual meeting of Japanese Geriatric Society were held in 1959 and the Japanese Gerontology Society meeting was also held in the same year by Geriatric Society and Geriatric Sociology Association. It was held every two years, and now it consists of 6 associations, that is Geriatrics. Geriatric Sociology. Basic Gerontology, Geriatric Dentistry, Geriatric Psychiatry and Care Management Associations. I will introduce the activities of these associations, and also introduce the contribution of departments of geriatrics in Japanese universities. I will also show the system of medical examinations and treatment, the education curriculum for medical students and residency, and research composing of various theme such as basic gene therapy as well as the evaluation of terminal care for the elderly in our Department of Geriatrics, Nagoya University Graduate School of Medicine. Finally, I would like to propose future research plan between departments of geriatrics in Japanese Universities and the National Institute for Longevity Sciences, which will open next March.


BatMis: a fast algorithm for k-mismatch mapping.
Second-generation sequencing (SGS) generates millions of reads that need to be aligned to a reference genome allowing errors. Although current aligners can efficiently map reads allowing a small number of mismatches, they are not well suited for handling a large number of mismatches. The efficiency of aligners can be improved using various heuristics, but the sensitivity and accuracy of the alignments are sacrificed. In this article, we introduce Basic Alignment tool for Mismatches (BatMis)--an efficient method to align short reads to a reference allowing k mismatches. BatMis is a Burrows-Wheeler transformation based aligner that uses a seed and extend approach, and it is an exact method. Benchmark tests show that BatMis performs better than competing aligners in solving the k-mismatch problem. Furthermore, it can compete favorably even when compared with the heuristic modes of the other aligners. BatMis is a useful alternative for applications where fast k-mismatch mappings, unique mappings or multiple mappings of SGS data are required. BatMis is written in C/C++ and is freely available from http://code.google.com/p/batmis/


Atypical hemolytic uremic syndrome and acute tubular necrosis induced by complement factor B gene (CFB) mutation: A case report.
Atypical hemolytic uremic syndrome (aHUS) is an uncommon and serious disease that manifests hemolytic anemia, thrombocytopenia, and acute kidney injury. Genetic complement abnormalities have been shown to be responsible. Compared with the aHUS caused by other mutated genes, aHUS secondary to CFB mutation in adults is extremely rare. We report an adult with CFB mutation developing aHUS. A 56-year-old man was admitted for 4-day history of nausea and fatigue, anuria for 2 days, and unconsciousness for 10 hours. The patient presented with life-threatening anemia, thrombocytopenia, acute kidney injury, and nervous system abnormalities. The patient had schistocytes on the peripheral blood smear, increased lactate dehydrogenase (LDH), and plasma-free hemoglobin levels. The patient was later found to harbor a pathogenic variant in the CFB gene (C.1598A>G), and was diagnosed with aHUS and acute kidney injury. The patient was treated by plasmapheresis, continuous renal replacement therapy, blood transfusion, and anti-infective and antihypertensive treatment. After the treatment, the patient's consciousness returned to normal, and the hemoglobin, platelet, and serum creatinine recovered. The disease activity remained quiescent during the follow-up. A rare heterozygous variant c.1598A>G p.Lys 533Arg in the CFB gene, which was associated with adult-onset aHUS, was described and successfully treated. This case can help in understanding the early diagnosis and effective therapies of this rare disease.


Genome Editing Using CRISPR/Cas9 System in the Rice Blast Fungus.
Fast and flexible genome manipulation is a powerful strategy for an in-depth understanding of molecular mechanisms in biological research. In recent years, CRISPR/Cas9-mediated genome editing has been used as a reliable genome manipulation method in a broad range of biological research including studies of filamentous fungi. The CRISPR/Cas9 system comprises a single-guide RNA (sgRNA) and a Cas9 protein, and the Cas9/sgRNA complex catalyzes a DNA double-strand break at the desired genomic locus. This protocol describes a fundamental CRISPR/Cas9 methodology that includes the design of the target sequence, construction of the CRISPR/Cas9 expression vector, and transformation for genome editing in Pyricularia (Magnaporthe) oryzae. This allows efficient targeted gene disruption, base editing, and reporter gene knock-in without any additional modifications of the host components. This protocol would be suitable for applying other CRISPR/Cas technologies and various functional genomics in P. oryzae.


Plasmodium falciparum: allelic variation in the merozoite surface protein 1 gene in wild isolates from southern Vietnam.
Allelic variation in the Plasmodium falciparum merozoite surface protein 1 (MSP1) gene is expressed as an association of allelic types in variable blocks. In this study, a PCR strategy that can detect 24 different MSP1 association types was used to investigate allelic variation in the MSP1 gene. We identified 236 distinct association type clones in 136 wild isolates collected from southern Vietnam, analysis of which revealed that (1) recombination between two representative allelic types in the central part of the MSP1 gene did not exist, (2) frequency distribution of MSP1 association types did not differ in different population groups, and (3) particular MSP1 association types were predominant. Statistical analysis for the association of allelic types indicated significant, nonrandom associations between blocks 4 and 6 but not between blocks 2 and 4, and 2 and 6. These results suggest that selection operates in favor of particular MSP1 association types. In addition, direct sequencing of 31 isolates confirmed reported sequence substitutions in the C-terminal 19-kDa Cys-rich region of MSP1, supporting a notion of limited variations in this region, a strong vaccine candidate molecule.


Omental evisceration through an abdominal stab wound: is exploratory laparotomy mandatory?
The authors reviewed and analysed the records of 75 patients with omental and bowel evisceration as a result of abdominal stab wounds to discover whether omental prolapse makes laparotomy mandatory. Major intraabdominal injuries were found in 82.7% of the total group, 82.9% in those with omental protrusion and 82.5% in patients with bowel prolapse. The nature of the injuries and the incidence of multiple organ trauma were similar in the two groups. Negative laparotomy did not cause important morbidity. The authors conclude that omental herniation after stab wounds of the abdomen should routinely prompt exploratory celiotomy.


Clinical and genetic profile of catecholaminergic polymorphic ventricular tachycardia in Hong Kong Chinese children.
To report our experience in the management of catecholaminergic polymorphic ventricular tachycardia in Hong Kong Chinese children. This case series study was conducted in a tertiary paediatric cardiology centre in Hong Kong. All paediatric patients diagnosed at our centre with catecholaminergic polymorphic ventricular tachycardia from January 2008 to October 2014 were included. Ten patients (five females and five males) were identified. The mean age at presentation and at diagnosis were 11.0 (standard deviation, 2.9) years and 12.5 (2.8) years, respectively. The mean delay time from first presentation to diagnosis was 1.5 (standard deviation, 1.3) years. They presented with recurrent syncope and six patients had a history of aborted cardiac arrest. Four patients were initially misdiagnosed to have epilepsy. Catecholaminergic polymorphic ventricular tachycardia was diagnosed by electrocardiogram at cardiac arrest (n=2), or provocation test, either by catecholamine infusion test (n=6) or exercise test (n=2). Mutations of the RyR2 gene were confirmed in six patients. Nine patients were commenced on beta-blockers after diagnosis. Despite medications, three patients developed aborted or resuscitated cardiac arrest (n=2) and syncope (n=1). Left cardiac sympathetic denervation was performed in five patients and an implantable cardioverter defibrillator was implanted in another. There was no mortality during follow-up. Catecholaminergic polymorphic ventricular tachycardia should be considered in children who present with recurrent syncope during exercise or emotional stress. Despite beta-blocker treatment, recurrent ventricular arrhythmias occur and may result in cardiac arrest.


Pericardial Angiosarcoma: A Diagnostic Challenge.
Cardiac angiosarcomas are the most common malignant primary cardiac tumors accounting for 31% of all primary cardiac tumors. However, primary pericardial angiosarcomas are extremely rare and are associated with high mortality. A 41-year-old male with a past medical history of end-stage renal disease (ESRD) on hemodialysis, follicular thyroid carcinoma, hypertension, and asthma presented with recurrent pericardial effusions. Previously, different imaging modalities had shown small hemodynamically stable pericardial effusions with pericardial thickening. His pericardial effusion was attributed to his ESRD until this presentation. However, during the current admission, chest X-ray showed cardiomegaly with lobulated left heart border. Computed tomography (CT) and transthoracic echocardiogram showed an increased posterior complex pericardial effusion when compared to previous imaging. A pericardial window was created and the space was evacuated. Pericardial fluid cytology reported rare atypical cells, which were indeterminate for malignancy. Histopathology from the pericardial biopsy revealed fibrous tissue with cellular proliferation, consistent with an angiosarcoma. A positron emission tomography revealed findings of angiosarcoma (hypermetabolic pericardial soft tissue nodularity and complex pericardial fluid) limited only to the pericardium. Unfortunately, the angiosarcoma was deemed unresectable, and the patient underwent one cycle of chemotherapy with paclitaxel. The patient's hospital course was further complicated by myelosuppression from chemotherapy and disseminated intravascular coagulation. Therefore, no further chemotherapy was pursued. A repeat echocardiogram showed constriction of both ventricles with loculated pericardial effusion. The patient eventually transitioned to comfort care and passed away. Through this case report, we would like to highlight that multiple confounders can be present when considering the etiology of persistent pericardial effusions. We suggest contemplating alternate diagnosis, such as malignancy, and initiate aggressive work-up especially in young individuals with recurrent, unexplained pericarditis.


The use of video laryngoscopy outside the operating room: A systematic review.
This study aimed to describe how video laryngoscopy is used outside the operating room within the hospital setting. Specifically, we aimed to summarise the evidence for the use of video laryngoscopy outside the operating room, and detail how it appears in current clinical practice guidelines. A literature search was conducted across two databases (MEDLINE and Embase), and all articles underwent screening for relevance to our aims and pre-determined exclusion criteria. Our results include 14 clinical practice guidelines, 12 interventional studies, 38 observational studies. Our results show that video laryngoscopy is likely to improve glottic view and decrease the incidence of oesophageal intubations; however, it remains unclear as to how this contributes to first-pass success, overall intubation success and clinical outcomes such as mortality outside the operating room. Furthermore, our results indicate that the appearance of video laryngoscopy in clinical practice guidelines has increased in recent years, and particularly through the COVID-19 pandemic. Current COVID-19 airway management guidelines unanimously introduce video laryngoscopy as a first-line (rather than rescue) device.


Use of magnetic resonance imaging in stereotactic surgery. A survey of members of the American Society of Stereotactic and Functional Neurosurgery.
Members of the American Society of Stereotactic Functional Neurosurgery were surveyed to determine the current use of magnetic resonance imaging (MRI) for stereotactic coordinate determination. Of 137 respondents, 48% used MRI as the sole technique for some biopsy procedures. Of those performing functional surgery, more surgeons use MRI alone than CT alone for localization. Of those surgeons who used MRI alone for stereotactic surgery, 98.6% were satisfied with this method. Significant factors related to the use of MRI amongst individual surgeons included the performance of > or = 2 stereotactic surgeries per week, the performance of radiosurgery, and being in an academic practice (p < 0.0004). The increasing use of MRI in stereotactic surgery must be coupled with quality assurance testing from frame manufacturers, imaging manufacturers, and surgeons.


Randomized short-term trial of two autoCPAP devices versus fixed continuous positive airway pressure for the treatment of sleep apnea.
We evaluated the efficacy of two different continuous positive airway pressure devices with automatic mask pressure adjustment (autoCPAP) in comparison with fixed CPAP in treating obstructive sleep apnea syndrome in 29 patients. The mean (+/- SE) apnea-hypopnea index was 46 +/- 4 per hour and the Epworth score was 14.2 +/- 0.7. Patients were treated over three consecutive 1-month periods with three regimens in random order: an autoCPAP device responding to apnea-hypopnea and snoring, another autoCPAP device responding to snoring and changes in flow contour, and fixed CPAP at the 90th pressure percentile titrated by autoCPAP over 2 weeks. Allowed pressure in the autoCPAP mode was 4 to 15 cm H2O. At the end of each treatment period, symptoms, quality of life, vigilance, and nocturnal breathing disturbances were evaluated. All three treatment modalities improved symptoms, quality-of-life domains, and apnea-hypopnea index significantly and to a similar degree. Mean (+/- SE) maintenance-of-wakefulness time increased by 4.5 +/- 1.8, 6.0 +/- 1.5, and 6.1 +/- 1.4 minutes with DeVilbiss AutoAdjust LT, AutoSet T, and fixed-pressure CPAP, respectively (p<0.001 vs. baseline, p=not significant for comparisons among the three modalities). We conclude that both autoCPAP devices were equally effective as fixed-pressure CPAP in improving major outcomes during short-term therapy of sleep apnea.


[Role of antivenoms in the treatment of snake envenomation].
The production of antivenoms, which were long deemed ineffective, dangerous and difficult to use, has improved dramatically. These antibodies (immunoglobulin G) are now fragmented, purified and controlled for their quality, leading to significantly better safety and facilitating their emergency use. Envenomation can result in various syndromes depending on the snake species: Viperidae venoms are highly inflammatory, hemorrhagic and necrotising, while Elapidae venoms can cause fatal respiratory paralysis. However, some Viperidae venoms can lead to asphyxiation similar to that observed in Elapidae envenomation while, conversely, Elapidae bites may be complicated by hemorrhage or necrosis, thus complicating etiologic diagnosis. Symptomatic treatment is complex, often insufficient, and frequently associated with adverse events. In contrast, antivenoms neutralize the venom and accelerate its clearance, thus providing an etiological treatment for envenomation, particularly in remote healthcare facilities in developing countries. Current formulations consist of polyvalent antivenoms covering most of the venomous species present in a specific region. The main limitation is their high cost, and the priority should be to develop new treatment strategies, including more affordable antivenoms, especially in developing countries where they are most needed.


EFFECTS OF SERIAL INBREEDING ON FITNESS COMPONENTS IN IMPATIENS CAPENSIS.
Studies of inbreeding depression in wild plants customarily compare the fitness of outcrossed progeny to progeny derived from one generation of self-pollination. We compare levels of inbreeding depression in a greenhouse in two populations of jewelweed using progeny derived from random outcrosses, one generation of self-pollination, and three generations of selling. The progeny have expected inbreeding coefficients of, respectively, 0, 0.5, and 0.875. Seedling survivorship declined linearly with the level of inbreeding in both populations. Inbreeding also increased the variability of emergence date. Maternal family membership affected early seedling performance and often interacted significantly with the level of inbreeding. In contrast, path analyses reveal that inbreeding had both negative linear and positive quadratic direct effects on seed and final plant weight, causing the highly inbred progeny to outperform progeny derived from one generation of selfing. These results suggest either the rapid purging of deleterious alleles or diminishing epistasis among the loci affecting these characters. It is not clear why the loci affecting survival responded differently.


What's new in p53?
p53 is the main intrinsic factor inducing apoptosis by recognizing the external stimuli and activating the p53 responsive genes to an irreversible series of events. P53 activates the transcription of specific proapoptotic genes called p53 target genes. A growing number of p53 responsive genes have been identified and numerous studies have demonstrated that p53 proapoptotic factors such as Noxa, Puma and Perp play cell type specific roles in p53's mediated response to certain stimuli. Perp (p53 apoptosis effector related to PMP-22) is a direct proapoptotic target gene encoding a tetraspan protein. Perp is highly expressed in cells undergoing apoptosis compared to cells under G1 arrest and its overexpression is sufficient to cause cell death in fibroblasts. Noxa is another member of the preapoptotic p53 genes family. When expressed Noxa acts in a BH3 motif-dependent localization to mitochondria, causing structural changes, activation of caspase 9 and release of cytochrome c from mitochondria to cytosol. Puma (p53 mutant of apoptosis) is another critical mediator of p53-dependent apoptosis. P53 binds to Puma-promoter gene sites, leading to puma production. The mtCLIC, a member of intracellular chloride channels, is a cytoplasmic and mitochondrial protein positively regulated by p53. Caspase 10 is induced in p53-dependent manner leading to cellular apoptosis. Other newly announced factors are also involved in p53-regulated apoptosis such as brain-specific angiogenesis inhibitor-1 (BSAI1), MSOD and GPX genes. A global discussion on this topic is attempted in the present review article.


[A Case of Suspected Delayed Postoperative Malignant Hyperthermia].
Malignant hyperthermia occurred 10 hours after surgery in a 72-year-old man who had received emergency laparoscopic cholecystectomy for severe acute cholecystitis with cholelethiasis. He had a high fever (39.4 degrees C) with liver damage before surgery. Anesthesia was induced with propofol and fentanyl and maintained with sevoflurane and epidural block using ropivacaine. Rocuronium was used as a muscle relaxant During surgery, body temperature decreased by cooling the body surface, but tachycardia continued. Ten hours after surgery, body temperature increased to the maximum of 40.6 degrees C and he went into shock. Then another 10 hours later, he developed cardiac arrest He recovered, but 22 hours later, second cardiac arrest occurred. After his second recovery, dantrolene was administered and body temperature decreased. He had hypoxic brain damage, but was dischanged from the hospital after tracheostomy on the 150th hospital day. From his clinical course, especially decrease in body temperature by dantrolene, he was suspected to have developed malignant hyperthermia. We should consider malignant hyperthermia when patient had a severe high fever postoperatively.


Zoledronic acid protects against local and systemic bone loss in tumor necrosis factor-mediated arthritis.
Increased osteoclast activity is a key factor in bone loss in rheumatoid arthritis (RA). This suggests that osteoclast-targeted therapies could effectively prevent skeletal damage in patients with RA. Zoledronic acid (ZA) is one of the most potent agents for blocking osteoclast function. We therefore investigated whether ZA can inhibit the bone loss associated with chronic inflammatory conditions. Human tumor necrosis factor (TNF)-transgenic (hTNFtg) mice, which develop severe destructive arthritis as well as osteoporosis, were treated with phosphate buffered saline, single or repeated doses of ZA, calcitonin, or anti-TNF, at the onset of arthritis. Synovial inflammation was not affected by ZA. In contrast, bone erosion was retarded by a single dose of ZA (-60%) and was almost completely blocked by repeated administration of ZA (-95%). Cartilage damage was partly inhibited, and synovial osteoclast counts were significantly reduced with ZA treatment. Systemic bone mass dramatically increased in hTNFtg mice after administration of ZA, which was attributable to an increase in trabecular number and connectivity. In addition, bone resorption parameters were significantly lowered after administration of ZA. Calcitonin had no effect on synovial inflammation, bone erosion, cartilage damage, or systemic bone mass. Anti-TNF entirely blocked synovial inflammation, bone erosion, synovial osteoclast formation, and cartilage damage but had only minor effects on systemic bone mass. ZA appears to be an effective tool for protecting bone from arthritic damage. In addition to their role in antiinflammatory drug therapy, modern bisphosphonates are promising candidates for maintaining joint integrity and reversing systemic bone loss in patients with arthritis.


A growth curve analysis of emotion dysregulation as a mediator for violence in individuals with and without borderline personality disorder.
Borderline personality disorder (BPD) is characterized by significant interpersonal conflict, however the factors that contribute to violence among this population are not well known. Individuals with BPD and other severe mental illnesses were followed in the community for 30 weeks post-inpatient discharge. Emotion dysregulation data and detailed measures of violence were collected every 10 weeks, along with measures of antisocial characteristics. Latent growth curve analyses were used to depict individual trajectories of emotion dysregulation throughout the course of the study, and the degree to which changes in emotion dysregulation mediated the risk-enhancing effect of BPD on subsequent interpersonal violence was assessed. Results showed that emotion dysregulation is a significant longitudinal mediator of violent behavior among individuals with BPD, and may serve as the primary mechanism that enhances risk for violence among this population. Implications for modifying existing treatment protocols to reduce interpersonal violence in this population are discussed.


A phylogenetic approach to the Philippines endemic centipedes of the genus Scolopendra Linnaeus, 1758 (Scolopendromorpha, Scolopendridae), with the description of a new species.
The genus Scolopendra Linnaeus, 1758 is represented in the Philippines' fauna by five species, two of which are endemic. Mitochondrial DNA sequences of gene cytochrome c oxidase subunit I (COI) were obtained from six Scolopendra specimens belonging to two endemic species and a new one, described here as Scolopendra paradoxa Doménech sp. nov. These sequences were analyzed together with another forty-one sequences from GenBank, including additional species of Scolopendra and a few representatives of other Scolopendridae genera. Phylogenetic trees inferred from the COI analysis using maximum likelihood and neighbor joining showed the three Philippines Scolopendra endemic species as a polyphyletic group coherent with their respective morphologies, although the position of S. spinosissima Kraepelin, 1903 varied within the obtained trees. Species delimitation based on standard external morphological characters was also concordant with the observed genetic distances, monophyly and node support, confirming S. subcrustalis Kronmüller, 2009 and S. paradoxa sp. nov. as separate species also at the molecular level, while only the position of S. spinosissima could not be properly established with any of the statistical methods used. In addition, the male genitalia of the three studied species were found to lack gonopods and a penis. Remarks on the ultimate legs prefemoral spinous formula of S. spinosissima plus a key to the species of the genus Scolopendra in the Philippines are provided.


Prevalence of Potential and Clinically Relevant Statin-Drug Interactions in Frail and Robust Older Inpatients.
A significant proportion of older people are prescribed statins and are also exposed to polypharmacy, placing them at increased risk of statin-drug interactions. To describe the prevalence rates of potential and clinically relevant statin-drug interactions in older inpatients according to frailty status. A cross-sectional study of patients aged ≥65 years who were prescribed a statin and were admitted to a teaching hospital between 30 July and 10 October 2014 in Sydney, Australia, was conducted. Data on socio-demographics, comorbidities and medications were collected using a standardized questionnaire. Potential statin-drug interactions were defined if listed in the Australian Medicines Handbook and three international drug information sources: the British National Formulary, Drug Interaction Facts and Drug-Reax(®). Clinically relevant statin-drug interactions were defined as interactions with the highest severity rating in at least two of the three international drug information sources. Frailty was assessed using the Reported Edmonton Frail Scale. A total of 180 participants were recruited (median age 78 years, interquartile range 14), 35.0% frail and 65.0% robust. Potential statin-drug interactions were identified in 10% of participants, 12.7% of frail participants and 8.5% of robust participants. Clinically relevant statin-drug interactions were identified in 7.8% of participants, 9.5% of frail participants and 6.8% of robust participants. Depending on the drug information source used, the prevalence rates of potential and clinically relevant statin-drug interactions ranged between 14.4 and 35.6% and between 14.4 and 20.6%, respectively. In our study of frail and robust older inpatients taking statins, the overall prevalence of potential statin-drug interactions was low and varied significantly according to the drug information source used.


Visualization of vermilion degradation using pump-probe microscopy.
Here, we demonstrate the use of pump-probe microscopy for high-resolution studies of vermilion degradation. Vermilion (mostly α-HgS), an important red pigment used in historical paintings, blackens over time, and metallic Hg and β-HgS have been implicated as possible degradation products. Conventional analysis techniques have trouble differentiating α- and β-HgS with sufficiently high spatial resolution. However, pump-probe microscopy can differentiate metallic mercury, α- and β-HgS, and map each distribution on the microscopic scale. We studied artificial degradation of α-HgS; femtosecond-pulsed laser irradiation induces an irreversible phase shift of α- to β-HgS, in which the initial presence of β-HgS grains can increase the rate of conversion in their vicinity. Continuous ultraviolet exposure instead generates both liquid Hg and β-HgS, with a conversion rate that increases with elevated temperatures. Last, we reveal the presence of β-HgS as a natural degradation product in discolored vermilion layers in a 14th century Italian painting.


Per capita remuneration of dentists and the quality of dental services.
The aim of this study was to examine the effects of a per capita based remuneration system on the quality of dental care. The basis for the study was a natural experiment in the county of Østfold in Norway in which all public dental officers (n = 34) were given the opportunity to renegotiate their contract from a fixed salary to a combined per capita and fixed salary contract. Quality was assessed according to three criteria: number of preventive procedures, number of under-diagnosed carious lesions and number of untreated carious lesions. This information was selected from a random sample of patient records for 18-year-olds in the autumn of 1999 (base-line data) and at the end of 2006 (final data), altogether 20 records per dentist. At the end of the period of evaluation, 26 dentists were still eligible for inclusion in the study. The data were analysed both descriptively and using multilevel difference-in-difference regression models. The main finding was that the transition to a per capita remuneration system did not lead to under-diagnosis of carious lesions, under-treatment or less prevention. A per capita remuneration scheme did not lead to a fall in the quality of dental care. This result must be assessed taking into account that the experiment was carried out within a publicly financed and managed dental service. Also, the incentive effect of the per capita contract was relatively weak.


Impact of inactive empiric antimicrobial therapy on inpatient mortality and length of stay.
The consequences of inactive empiric antimicrobial therapy are not well-described and may cause prolonged hospitalization or infection-related mortality. In vitro susceptibility results for 884 patients hospitalized at an academic medical center with gram-negative bloodstream infections (GNBI) from 2001 to 2003 were matched to antimicrobial orders within 24 h of culture. Clinical characteristics, organism, inpatient mortality, and length of stay after culture for patients with GNBI were compared between patients receiving active versus inactive empiric antimicrobial therapy. A total of 14.1% of patients with GNBI received inactive empiric therapy, defined as no antimicrobial therapy within 24 h of the culture active against the identified organism based on in vitro microbiology reports. Patients who received inactive therapy were more likely to be younger, to be infected with Pseudomonas aeruginosa, to have a nosocomial infection, and to receive antimicrobial monotherapy but less likely to be bacteremic with Escherichia coli or to have sepsis (P < 0.05). There were no significant differences in mortality between patients receiving active versus inactive empiric therapy (16.1% versus 13.6%, respectively) or in length of stay after positive culture (11.5 days versus 12.6 days, respectively). Only 45 patients had greater than 2 days of exposure to inactive therapy; however, 8/30 patients (26.7%) who never received active antimicrobial therapy died while in the hospital. Inactive empiric therapy was more common in healthier patients. Inactive antimicrobial therapy in the first 24 h did not significantly impact average outcomes for GNBI among hospitalized patients but may have caused harm to specific individuals.


Dynamic treatment of vibrational energy relaxation in a heterogeneous and fluctuating environment.
A computational approach to describe the energy relaxation of a high-frequency vibrational mode in a fluctuating heterogeneous environment is outlined. Extending previous work [H. Fujisaki, Y. Zhang, and J. E. Straub, J. Chem. Phys. 124, 144910 (2006)], second-order time-dependent perturbation theory is employed which includes the fluctuations of the parameters in the Hamiltonian within the vibrational adiabatic approximation. This means that the time-dependent vibrational frequencies along a molecular dynamics trajectory are obtained via a partial geometry optimization of the solute with fixed solvent and a subsequent normal mode calculation. Adopting the amide I mode of N-methylacetamide in heavy water as a test problem, it is shown that the inclusion of dynamic fluctuations may significantly change the vibrational energy relaxation. In particular, it is found that relaxation occurs in two phases, because for short times (approximately < 200 fs) the spectral density appears continuous due to the frequency-time uncertainty relation, while at longer times the discrete nature of the bath becomes apparent. Considering the excellent agreement between theory and experiment, it is speculated if this behavior can explain the experimentally obtained biphasic relaxation the amide I mode of N-methylacetamide.


Determination of the elongate spermatid-Sertoli cell ratio in various mammals.
Criteria were devised for determining the elongate spermatid-Sertoli cell ratio in various mammalian species at the electron microscope level. When data from particular species were pooled, the values were: rabbit, 12.17:1, hamster, 10.75:1; gerbil, 10.64:1; rat, 10.32:1; guinea-pig, 10.10:1; vole, 9.75:1; and monkey, 5.94:1. The elongate spermatid-Sertoli cell ratio is a measure of the workload of the Sertoli cell and is a prime factor determining their efficiency. The higher the ratio, the higher the sperm output is likely to be per given weight of seminiferous tubule parenchyma for a particular species.


Purification and sequence of rat oxyntomodulin.
Structural information about rat enteroglucagon, intestinal peptides containing the pancreatic glucagon sequence, has been based previously on cDNA, immunologic, and chromatographic data. Our interests in testing the physiological actions of synthetic enteroglucagon peptides in rats required that we identify precisely the forms present in vivo. From knowledge of the proglucagon gene sequence, we synthesized an enteroglucagon C-terminal octapeptide common to both proposed enteroglucagon forms, glicentin and oxyntomodulin, but sharing no sequence overlap with glucagon. We then developed a radioimmunoassay using antibodies raised against the octapeptide that was specific for enteroglucagon peptides without cross-reacting with glucagon. Rat intestine was extracted, and one presumptive enteroglucagon form was purified by following the enteroglucagon C-terminal octapeptide-like immunoreactivity through several HPLC purification steps. Structural characterization of the material by amino acid composition, microsequence, and mass spectral analyses identified the peptide as rat oxyntomodulin. The 37-residue peptide consists of pancreatic glucagon plus the C-terminal extension, Lys-Arg-Asn-Arg-Asn-Asn-Ile-Ala. This now permits synthesis of an unambiguous duplicate of endogenous rat oxyntomodulin for physiological studies.


Influence of neutral protease activity on human islet isolation outcome.
Observations in rat pancreata have revealed that enzymatic islet release is mediated by both collagenase and neutral protease (NP), a critical effector of islet integrity. Since no information is available about the effect of NP activity on islet release from the human pancreas, the present study evaluated the effect of various NP concentrations on the outcome of human islet isolation. Following intraductal collagenase distension, pancreata obtained from adult multiorgan donors were digested using 2000 PZ-U of purified Serva collagenase NB 1 supplemented with 2.6 (n = 10) or 4.5% (DMC-U/PZ-U) (n = 10) of NP. Increasing NP from 2.6% to 4.5% reduced the amount of undigested tissue from 22 +/- 2 to 17 +/- 2 g (P < .05) while simultaneously increasing the volume of digested tissue (26 +/- 2 vs 40 +/- 3 mL, P < .01). Increased NP concentrations increased the islet yield prepurification (459,800 +/- 22,900 vs 587,600 +/- 69,000 IEQ, P < .05), but simultaneously affected islet purification, resulting in equal islet yields (345,700 +/- 31,200 vs 391,500 +/- 35,400 IEQ, NS) and less purity (70 +/- 6 vs 49% +/- 5%, P < .01). A NP concentration of 4.5% reduced the stimulation index (4.7 +/- 1.2 vs 2.0 +/- 0.5, P < .01) and viability (100 +/- 1 vs 95% +/- 3%, P < .05). Although increased NP activity seems to improve islet release from adult human pancreata, it significantly affects islet viability and function. The reduction in purity reflected damage to acinar tissue by increased NP activity presumably affecting islet integrity.


Entrainment of atrioventricular nodal reentrant tachycardias during overdrive pacing from high right atrium and coronary sinus. With special reference to atrioventricular dissociation and 2:1 retrograde block during tachycardias.
Entrainment was attempted during electrophysiologic evaluation of 8 patients with atrioventricular (AV) nodal reentrant tachycardia. Entrainment could be performed while pacing from the high right atrium in 35 of 35 episodes, from proximal coronary sinus in 9 of 21 episodes and from distal coronary sinus in 10 of 20 episodes. The minimal rates required were 8 to 40 beats/min faster than those of the tachycardias. That the atria (as defined in electrophysiologic studies) were not a necessary component of the reentry circuit was suggested by the occurrence, during tachycardia, of short episodes of AV dissociation and of 1 episode of 2:1 retrograde block. For the tachycardia to be interrupted, the pacing rate usually had to be slightly faster than that required to entrain, as well as sufficiently rapid to produce anterograde block of an atrial impulse in the slow AV nodal pathway. Moreover, termination of tachycardia apparently was a function of the pacing site. In some episodes, either because of a proximity effect or because of a preferential input into the upper common pathway, coronary sinus pacing terminated the tachycardia at slower rates or with fewer stimuli than high right atrial pacing. Thus, patients with drug-resistant AV nodal reentrant tachycardias may benefit from recently introduced pacing techniques for termination of tachycardia through entrainment.


CD4 cell priming and tolerization are differentially programmed by APCs upon initial engagement.
Bone marrow-derived APCs present both parenchymal-self and pathogen-derived Ags in a manner that elicits either T cell tolerization or immunity, respectively. To study the parameters that confer tolerogenic vs immunogenic APC function we used an adoptive transfer system in which naive TCR transgenic hemagglutinin (HA)-specific CD4(+) T cells are either tolerized upon encountering HA expressed constitutively as a parenchymal self-Ag (self-HA) or primed to express effector function upon encountering transiently expressed vaccinia-derived HA (viral-HA). When the duration of viral-HA presentation was extended for the period required to elicit tolerization toward self-HA, CD4 cell tolerization to viral-HA did not occur. Furthermore, CD4 cells exhibited both phenotypic as well as functional differences during early stages of tolerization and priming, suggesting that these divergent differentiation processes are programmed soon after the initial APC-CD4 cell interaction. When mice expressing self-HA were infected with an irrelevant vaccinia, CD4 cell tolerization still occurred, indicating that priming vs tolerization cannot be explained by pathogen-induced third parties (i.e., non-APCs) that act directly on CD4 cells. Taken together, these results suggest that CD4 cell tolerization to parenchymal self-Ags and priming to pathogen-derived Ags are initiated by functionally distinct APCs.


A single mutation in P450BM-3 enhances acyl homoserine lactone: acyl homoserine substrate binding selectivity nearly 250-fold.
Quorum sensing is the process by which bacteria alter gene regulation in response to their population density. The enzymatic inactivation of quorum signals has shown promise for use in genetically modified organisms resistant to pathogens. We recently characterized the ability of a cytochrome P450, P450BM-3, to oxidize the quorum sensing signals known as acyl homoserine lactones. The oxidation of the acyl homoserine lactones reduced their activity as quorum signals. The enzyme also oxidized the inactive lactonolysis products, acyl homoserines. The enzyme showed similar binding affinity for the acyl homoserine lactones and acyl homoserines. The latter reaction may lead to problems when lactonases and the P450-dependent system are used in tandem, as oxidation of the acyl homoserines produced by lactonolysis in vivo may compete with acyl homoserine lactone oxidation by the cytochrome P450. We report here that a single mutation (R47S) in P450BM-3 is capable of increasing the acyl homoserine lactone: acyl homoserine substrate binding selectivity of the enzyme nearly 250-fold, reducing the potential for competition by acyl homoserines and significantly enhancing the potential for use of P450BM-3 as part of a pathogen resistance system in genetically modified crops.


Synthesis and stability of soluble hexacenes.
The synthesis of new silylethyne-substituted hexacene derivatives to investigate their solubility, stability, and pi-stacking is reported. It was found that "butterfly" dimerization, rather than photooxidation, is the dominant decomposition pathway for these molecules and that stability can be enhanced by functionalization to prevent close contact between specific regions of the aromatic core. Dimerization regioselectivity can be altered by suitable engineering of the solid-state arrangement of the chromophores.


Heidelberg's first experiences in establishing a living-donation liver transplantation programme.
Since indications for liver transplantation (LTx) have increased and the shortage of brain dead transplant organs continues, an increase in deaths related to extended waiting periods can be documented. Living-related LTx (LRLTx) is a good alternative that can help compensate the shortness of organs for transplantation and can reduce waiting periods and deaths related to a long waiting list. We started a LRLTx programme at the transplantation centre, Department of Surgery at the University of Heidelberg to increase our transplantation activities. In this article we present our primary experiences and protocol. From September 2002 to June 2003, three LRLTx were performed. Indications in recipients were hepatocellular carcinoma in two cases and liver cirrhoses due to hepatitis B in one patient. Two patients received the right liver (segments V-VIII) of their donors. One patient received the complete liver collected as a Domino LTx. All liver transplants were performed without the use of a venous bypass. The peri- and post-operative courses of our donors were without complications, except for an infected intraabdominal bilioma, which was drained interventionally. The donors were fully able to work after the operation. The coordinative and logistical work should not be underestimated. This therapy should only be offered and performed in centres with excellent expert knowledge in liver and transplantation surgery.


p90 RSK-1 associates with and inhibits neuronal nitric oxide synthase.
Evidence is presented that RSK1 (ribosomal S6 kinase 1), a downstream target of MAPK (mitogen-activated protein kinase), directly phosphorylates nNOS (neuronal nitric oxide synthase) on Ser847 in response to mitogens. The phosphorylation thus increases greatly following EGF (epidermal growth factor) treatment of rat pituitary tumour GH3 cells and is reduced by exposure to the MEK (MAPK/extracellular-signal-regulated kinase kinase) inhibitor PD98059. Furthermore, it is significantly enhanced by expression of wild-type RSK1 and antagonized by kinase-inactive RSK1 or specific reduction of endogenous RSK1. EGF treatment of HEK-293 (human embryonic kidney) cells, expressing RSK1 and nNOS, led to inhibition of NOS enzyme activity, associated with an increase in phosphorylation of nNOS at Ser847, as is also the case in an in vitro assay. In addition, these phenomena were significantly blocked by treatment with the RSK inhibitor Ro31-8220. Cells expressing mutant nNOS (S847A) proved resistant to phosphorylation and decrease of NOS activity. Within minutes of adding EGF to transfected cells, RSK1 associated with nNOS and subsequently dissociated following more prolonged agonist stimulation. EGF-induced formation of the nNOS-RSK1 complex was significantly decreased by PD98059 treatment. Treatment with EGF further revealed phosphorylation of nNOS on Ser847 in rat hippocampal neurons and cerebellar granule cells. This EGF-induced phosphorylation was partially blocked by PD98059 and Ro31-8220. Together, these data provide substantial evidence that RSK1 associates with and phosphorylates nNOS on Ser847 following mitogen stimulation and suggest a novel role for RSK1 in the regulation of nitric oxide function in brain.


A case of refractory immune checkpoint inhibitor-induced colitis with Clostridioides difficile infection.
The clinical symptoms of an immune checkpoint inhibitor (ICI)-induced colitis are similar to those of ulcerative colitis. ICI-induced colitis, like ulcerative colitis, may be complicated by other colitis, such as Clostridioides difficile infection (CDI). A 72-year-old man was admitted because of watery and bloody stools 10 times a day after three courses of nivolumab (antibodies against programmed death 1) and ipilimumab (cytotoxic T-lymphocyte-associated antigen-4) for stage IV renal cell carcinoma. Colonoscopy revealed erythema and multiple erosions in the colon. Histopathological examination of colonic mucosa revealed diffuse inflammatory cell infiltration and apoptosis. The initial cytomegalovirus antigen test and C. difficile detection assay results were negative. Based on these findings, we diagnosed the patient with ICI-induced colitis and discontinued ICI therapy. The symptoms did not improve despite the administration of Prednisolone and infliximab. A repeat colonoscopy revealed a new appearance of pseudomembranes from the sigmoid colon to the rectum one month after the start of these treatments. At this point, the patient tested positive for C. difficile. With treatment with vancomycin for CDI, the abdominal symptoms gradually decreased. Nivolumab alone was cautiously restarted. However, no colitis recurrence and further tumor reduction were observed. Here, we report our experience of a case of refractory ICI-induced colitis complicated by CDI. ICI-induced colitis may be complicated by CDI and should be carefully treated with repeated CDI testing if refractory to treatment. We believe that our observation will provide helpful information for determining an appropriate treatment strategy for ICI-induced colitis.


Combined prognostic value of peak O(2) uptake and microvolt level T-wave alternans in patients with idiopathic dilated cardiomyopathy.
Despite the great improvement in clinical management of patients with idiopathic dilated cardiomyopathy (DCM), sudden cardiac death (SCD) and death due to worsening heart failure (HF) remain a challenging problem. The assessment of oxygen consumption (peakVO(2)) has been recognized as an independent marker of mortality. Nevertheless peakVO(2) is not helpful in the risk stratification of SCD. Given this limitation, the association with another non-invasive test able to predict SCD such as microvolt level T-wave alternans (MTWA) would be useful. To determine the combined predictive value of peakVO(2) and MTWA in patients with DCM. Seventy consecutive DCM patients were prospectively investigated. PeakVO(2) and MTWA were determined during bicycle exercise testing. Primary composite study end-point was defined as major cardiac events (MCE): total cardiac death or documented sustained VT/VF (including appropriate ICD shock). Secondary end-point was defined as arrhythmic events (AE): SCD or documented sustained VT/VF. Thirty-nine patients (55%) had a peakVO(2)<10 ml/kg/min, while 40 patients (57%) showed an abnormal MTWA test. During an average follow-up of 19.2+/-10.7 months, 11 MCE of which 6 AE have been documented. Among patients with abnormal MTWA and peakVO(2)<10 ml/kg/min 8 MCE of which 5 AE occurred while among patients with normal MTWA and peakVO(2)> or =10 ml/kg/min no event occurred. From multivariate analysis, the combined prognostic value of MTWA and peakVO(2) achieved statistical significance for MCE (p=0.03, HR 0.28, 95% CI 0.12-0.95) and for AE (p=0.05, HR 0.39, 95% CI 0.18-0.99) while MTWA alone was a significant predictor of AE (p=0.04, HR 0.32, 95% CI 0.14-0.93). Our results suggest that only the association of MTWA and peakVO(2), but not the two single tests, is a significant prognostic marker of both MCE and AE in DCM patients. However, MTWA alone confirms its predictive power as arrhythmic risk stratifier in this population.


Do place-of-death preferences for patients receiving specialist palliative care change over time?
Discussing preferred place of death (PPD) with patients approaching end of life is an important part of anticipatory care planning. Preferences at a specific point in time have been described; however the extent to which preferences may change is unclear. This study examines changes in PPD. A retrospective case note review of all patients who died under the care of a specialist palliative care service during a 6-month period in 2012 was undertaken. Notes relating to 299 patients were examined by a member of the clinical team. Of the 204 patients who had more than one PPD assessment, 57% showed a change in preference status between the first and last assessment. The majority changed from an unclarified preference to identifying a preferred place. Only 15% of patients with two or more assessments switched from one location to another. Most patients under the care of a specialist palliative care service identify a preference for place of death as end of life approaches. Only a minority change their preference once a preferred place has been elicited. We recommend that patients are supported to explore their preferences for PPD as part of specialist palliative care, and that preferences are reviewed as end of life approaches.


Amantadine Attenuated Hypoxia-Induced Mitochondrial Oxidative Neurotoxicity, Apoptosis, and Inflammation via the Inhibition of TRPM2 and TRPV4 Channels.
The hypoxia (HPX) acts the brain injury and apoptosis via the Ca2+ influx-induced excessive mitochondria free reactive oxygen species (mitROS) in neurons. The effective treatment of HPX is not possible yet. In addition to the antiviral and antiparkinsonian actions, amantadine (AMN) has been evaluated as a drug in treatments against brain injury. TRPM2 and TRPV4 channels are activated by mitROS. AMN attenuates NMDA receptor-induced Ca2+ influx, mitROS, inflammation, and apoptosis in the brain. However, the molecular pathways underlying AMN's neuroprotection against HPX remain elusive. We investigated the protective role of AMN via attenuation of TRPM2 and TRPV4 on oxidative neurotoxicity, mitochondrial membrane potential (ΔΨm), inflammation, and apoptosis in neuronal cells (SH-SY5Y). The SH-SY5Y and HEK293 cells were divided into six groups as follows: control, AMN (750 µM for 48 h), HPX (200 µM CoCl2 for 24 h), HPX + AMN, HPX + TRPM2 blockers (25 µM ACA or 100 µM 2APB for 30 min), and HPX + TRPV4 blocker (ruthenium red (RuR)-1 µM for 30 min). The HPX caused to upregulation of Ca2+ influx with an upregulation of ΔΨm and mitROS. The changes were not observed in the absence of TRPM2 and TRPV4 in the HEK293 cells. When HPX induction, TRPV4 agonist (GSK1016790A) and TRPM2 agonists (ADP-ribose and H2O2)-induced channel activity were diminished by the incubation of AMN and channel antagonists (RuR, ACA, and 2APB). The changes of mitROS, apoptotic markers (caspase-3 and -9), cell death rate, cell viability, cytokine (IL-1β, IL-6, and TNF-α), ΔΨm, and Zn2+ concentrations were also restored by the incubation of AMN. In conclusion, the treatment of AMN attenuated HPX-mediated mitROS, apoptosis, and TRPM2/TRPV4-mediated overload Ca2+ influx and may provide an avenue for protecting the HPX-mediated neurodegenerative and cerebrovascular diseases associated with the upregulation of mitROS, Ca2+, and Zn2+ concentration.


[Osteoarthritis of the hip].
If all the etiological factors are taken into consideration, idiopathic coxarthrosis is less prevalent than formerly. Three factors are new or reappraised: sport practice (more than 10 years in competition), certain occupations (dancer, heavy load worker) and heritability. Three are classical and yet present: developmental dysplasias, acquired dysmorphias and traumas. Hip osteoarthritis should be followed up with the help of pain intensity measured on visual analog scale, algofunctional index and self-assessment of handicap. These 3 parameters--and not the X-ray image--are the basis of the decision for total hip replacement. However, the latter could be foreseen as necessary within the next 1-4 years when a rapid joint space narrowing is observed (about 5-7% of cases).


What is the role of controls in an outpatient department on progression of renal disease?
Evidence that protein restriction retards the progression of chronic renal failure in man is mostly derived from anecdotic data and retrospective studies with inappropriate controls and inferior methodology for evaluating renal function. Data from a prospective randomized study in Stockholm demonstrate that more frequent clinical follow-ups without changing the dietary intake of protein significantly retards the progression of renal failure and that the degree of retardation is correlated to improvement in blood pressure control.


Factors Associated with Children's Physical Activity During Youth Soccer Practices.
Purpose: To identify practice and social contextual factors that associate with physical activity (PA) levels of children during their participation in a youth soccer program. Methods: Twenty-seven youth soccer teams serving children ages 6-11 years participated. Research staff directly observed and recorded PA intensity and practice and social contextual factors using momentary time-sampling procedures. Each team was observed for 1 practice, during which approximately 6 children were each observed for twenty 30-s observation blocks (10-s observation, 20-s recording). In total, children were observed for 3,102 intervals. Multilevel logistic regression analyses were conducted to describe associations between PA intensity and practice and social contexts. Interaction terms were introduced into the models to determine if the associations differed across girls-only, boys-only, and coed teams. Results: A total of 158 children were observed across the 27 teams. Children were more likely to engage in moderate or vigorous PA while performing fitness (Odds Ratio [OR], 9.9, 95% CI = 5.34-18.04), game (OR, 4.0, 95% CI = 2.88-5.66), warm-up (OR, 2.8, 95% CI = 1.85-4.11), and drill (OR, 1.9, 95% CI = 1.41-2.67) activities compared to tactic/instructional activities. The associations between PA intensity levels and practice and social contexts did not differ across girls-only, boys-only, and coed teams. Conclusions: Fitness activities and full-team game play were associated with higher PA intensity levels during children's participation in youth soccer practices. Youth sport practice protocols can be modified to increase children's physical activity.


A meta-analysis of the relative sensitivity of neuropsychological screening tests.
The National Institute of Mental Health recently formed a committee of experts which published a proposal for a lengthy neuropsychological test battery judged most apt to detect diffuse brain damage while covering a wide range of cognitive abilities. The purposes of the present study were (1) to assemble empirical evidence of the existence of statistically significantly sensitive screening tests across an equivalently wide range of functional domains, and (2) to present this evidence systematically, in such a manner as to help clinicians select subsets of screening tests manifesting greatest sensitivity to diffuse brain damage. It was found that in certain functional domains (speed of processing, problem solving, executive functions), marked differences in sensitivity occurred. Brief tests just as sensitive as time-consuming tests were also identified. The reader is cautioned about possible extraneous sources of the differences obtained (test and group selection bias, test reliability, test difficulty, procedural effects, Type I error). Nevertheless, it was concluded that a brief highly sensitive and functionally wide-ranging neuropsychological test battery for screening cases of putative diffuse brain dysfunction can be assembled.


New methods for semiconductor charge-diffusion-length measurements using synchrotron radiation.
The extension of a new theory on the X-ray energy response of semiconductor detectors is carried out to characterize the X-ray response of a multichannel semiconductor detector fabricated on one silicon wafer. Recently, these multichannel detectors have been widely utilized for position-sensitive observations in various research fields, including synchrotron radiation research and fusion-plasma investigations. This article represents the verification of the physics essentials of a proposed theory on the X-ray response of semiconductor detectors. The three-dimensional charge-diffusion effects on the adjoining detector-channel signals are experimentally demonstrated at the Photon Factory for two types of multichannel detectors. These findings are conveniently applicable for measuring diffusion lengths for industrial requirements.


A CACTA-like transposable element in the upstream region of BnaA9.CYP78A9 acts as an enhancer to increase silique length and seed weight in rapeseed.
Rapeseed (Brassica napus L.) is a model plant for polyploid crop research and the second-leading source of vegetable oil worldwide. Silique length (SL) and seed weight are two important yield-influencing traits in rapeseed. Using map-based cloning, we isolated qSLWA9, which encodes a P450 monooxygenase (BnaA9.CYP78A9) and functions as a positive regulator of SL. The expression level of BnaA9.CYP78A9 in silique valves of the long-silique variety is much higher than that in the regular-silique variety, which results in elongated cells and a prolonged phase of silique elongation. Plants of the long-silique variety and transgenic plants with high expression of BnaA9.CYP78A9 had a higher concentration of auxin in the developing silique; this induced a number of auxin-related genes but no genes in well-known auxin biosynthesis pathways, suggesting that BnaA9.CYP78A9 may influence auxin concentration by affecting auxin metabolism or an unknown auxin biosynthesis pathway. A 3.7-kb CACTA-like transposable element (TE) inserted in the 3.9-kb upstream regulatory sequence of BnaA9.CYP78A9 elevates the expression level, suggesting that the CACTA-like TE acts as an enhancer to stimulate high gene expression and silique elongation. Marker and sequence analysis revealed that the TE in B. napus had recently been introgressed from Brassica rapa by interspecific hybridization. The insertion of the TE is consistently associated with long siliques and large seeds in both B. napus and B. rapa collections. However, the frequency of the CACTA-like TE in rapeseed varieties is still very low, suggesting that this allele has not been widely used in rapeseed breeding programs and would be invaluable for yield improvement in rapeseed breeding.


EEGIFT: group independent component analysis for event-related EEG data.
Independent component analysis (ICA) is a powerful method for source separation and has been used for decomposition of EEG, MRI, and concurrent EEG-fMRI data. ICA is not naturally suited to draw group inferences since it is a non-trivial problem to identify and order components across individuals. One solution to this problem is to create aggregate data containing observations from all subjects, estimate a single set of components and then back-reconstruct this in the individual data. Here, we describe such a group-level temporal ICA model for event related EEG. When used for EEG time series analysis, the accuracy of component detection and back-reconstruction with a group model is dependent on the degree of intra- and interindividual time and phase-locking of event related EEG processes. We illustrate this dependency in a group analysis of hybrid data consisting of three simulated event-related sources with varying degrees of latency jitter and variable topographies. Reconstruction accuracy was tested for temporal jitter 1, 2 and 3 times the FWHM of the sources for a number of algorithms. The results indicate that group ICA is adequate for decomposition of single trials with physiological jitter, and reconstructs event related sources with high accuracy.


'Come share my world' of 'madness': a systematic review of clinical, diagnostic and therapeutic aspects of folie à deux.
Folie à deux, also known as shared psychotic disorder, shows intrinsic differences when compared to other psychiatric disorders. Literature about this condition is scanty and findings about its clinical correlates are not univocal. The present systematic review aimed at critically summarizing the existing evidence about folie à deux, also focussing on psychopathological, diagnostic and treatment features. The electronic databases PUBMED, Web of Science and Scopus were systematically searched according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Articles reporting original data about socio-demographic, aetiological, clinical, diagnostic and treatment features of folie à deux, providing information about both primaries and secondaries, were included. Risk of bias was evaluated using the Critical Appraisal Skills Programme (CASP) instruments. The initial search yielded 834 records. After the screening process only 9 papers were deemed eligible for inclusion in the review. Folie à deux emerged to be a heterogeneous condition with a complex etiopathogenesis. The clinical presentation of the disorder included a multi-facet aspect, going beyond the classical description mainly focussed on delusions and possibly explaining conflicting outcomes of different treatments. Diagnostic categories appear to be often reductive and the need for a dimensional approach capable of global reliability emerged.


[Obstetrical complications of antiphospholipid syndrome].
Spontaneous miscarriages and fetal deaths take part of the definition of the antiphospholipid syndrome. Preeclampsia, fetal distress, preterm delivery, intrauterine growth retardation, mother's or newborn's vascular thrombosis are also included in the spectrum of clinical events linked to the antiphospholipid antibodies in pregnant women. The pathogenesis is not fully understood, but involves interactions between antibodies and phospholipidic molecules that are complexed to plasmatic proteins of the coagulation, or present on endothelial cell and platelet surfaces. The poor spontaneous prognosis of pregnancy in the context of the antiphospholipid syndrome is dramatically improved by preventive treatment. Therapeutic options are not codified and are various combinations of low-dose aspirin, heparin, steroids, intravenous immunoglobulins, and fish-oil derivatives. Excellent obstetrical care is mandatory in every case.


Recapitulating essential pathophysiological characteristics in lung-on-a-chip for disease studies.
Lung diseases have become a significant challenge to public healthcare worldwide, which stresses the necessity of developing effective biological models for pathophysiological and pharmacological studies of the human respiratory system. In recent years, lung-on-a-chip has been extensively developed as a potentially revolutionary respiratory model paradigm with high efficiency and improved accuracy, bridging the gap between cell culture and preclinical trials. The advantages of lung-on-a-chip technology derive from its capabilities in establishing 3D multicellular architectures and dynamic microphysiological environments. A critical issue in its development is utilizing such capabilities to recapitulate the essential components of the human respiratory system for effectively restoring physiological functions and illustrating disease progress. Here we present a review of lung-on-a-chip technology, highlighting various strategies for capturing lung physiological and pathological characteristics. The key pathophysiological characteristics of the lungs are examined, including the airways, alveoli, and alveolar septum. Accordingly, the strategies in lung-on-a-chip research to capture the essential components and functions of lungs are analyzed. Recent studies of pneumonia, lung cancer, asthma, chronic obstructive pulmonary disease, and pulmonary fibrosis based on lung-on-a-chip are surveyed. Finally, cross-disciplinary approaches are proposed to foster the future development of lung-on-a-chip technology.


Laparoscopic splenectomy and porto-azygos disconnection: clinical research in the treatment of portal hypertension.
To investigate the safety and efficacy of laparoscopic splenectomy and portaazygous devascularization, we studied laparoscopic splenectomy and porto-azygos devascularization patients within the peri-operative period. Clinical data and curative effect are detailed alongside statistical analysis. The laparoscopic splenectomy and porto-azygos devascularization operation time was 2.56 + 0.62 hours. The intraoperative bleeding and anal exhaust time was 149.5 + 32.7ml 3.47 + 1.32 days, and the hospitalization time was 5.05 + 1.22 days. When the spleen volume was greater than or equal to 1.5 liters, the rate of open abdominal surgery increased significantly. After 1, 2, 3, and 4 years of follow-up, cumulative recurrence bleeding rates were 0, 5.20%, 9.98%, and 15.83%, respectively. Laparoscopic splenectomy and pericardial devascularization is safe, effective, and feasible, and it can be confirmed by enhanced spiral computed tomography (CT). Whether spleen volume greater than 1.5L is suited to laparoscopic surgery requires further research.


Predictive Value of the CHA2DS2-VASc Score for Mortality in Hospitalized Acute Coronary Syndrome Patients With Chronic Kidney Disease.
Chronic kidney disease (CKD) patients have a high prevalence of coronary artery disease and a high risk of cardiovascular events. The present study assessed the value of the CHA2DS2-VASc score for predicting mortality among hospitalized acute coronary syndrome (ACS) patients with CKD. This was a retrospective cohort study that included CKD patients who were hospitalized for ACS from January 2015 to May 2020. The CHA2DS2-VASc score for each eligible patient was determined. Patients were stratified into two groups according to CHA2DS2-VASc score: <6 (low) and ≥6 (high). The primary endpoint was all-cause mortality. A total of 313 eligible patients were included in the study, with a mean CHA2DS2-VASC score of 4.55 ± 1.68. A total of 220 and 93 patients were assigned to the low and high CHA2DS2-VASc score groups, respectively. The most common reason for hospitalization was unstable angina (39.3%), followed by non-ST-elevation myocardial infarction (35.8%) and ST-elevation myocardial infarction (24.9%). A total of 67.7% of the patients (212/313) received coronary reperfusion therapy during hospitalization. The median follow-up time was 23.0 months (interquartile range: 12-38 months). A total of 94 patients (30.0%) died during follow-up. The high score group had a higher mortality rate than the low score group (46.2 vs. 23.2%, respectively; p < 0.001). The cumulative incidence of all-cause death was higher in the high score group than in the low score group (Log-rank test, p < 0.001). Multivariate Cox regression analysis indicated that CHA2DS2-VASc scores were positively associated with all-cause mortality (hazard ratio: 2.02, 95% confidence interval: 1.26-3.27, p < 0.001). The CHA2DS2-VASc score is an independent predictive factor for all-cause mortality in CKD patients who are hospitalized with ACS. This simple and practical scoring system may be useful for the early identification of patients with a high risk of death.


Hypoglycemia and Dementia.
Recently, cardiovascular morbidity and mortality have been continuously reduced in people with diabetes through the intensive management of multiple risk factors. However, contrary to this trend, the prevalence and clinical significance of neurodegenerative disorders are steadily increasing. In previous studies, diabetes mellitus has been found to play an important role in increasing the risk of dementia and cognitive dysfunction through various mechanisms. In particular, hypoglycemia is a frequent clinical situation in the strict management of diabetes and is well known as an important factor that directly affects the risk of dementia and cognitive impairment. This article describes the occurrence of hypoglycemia in patients with diabetes, and summarizes the resultant risks of dementia and cognitive dysfunction.


Synaptoproteomic analysis of a rat gene-environment model of depression reveals involvement of energy metabolism and cellular remodeling pathways.
Major depression is a severe mental illness that causes heavy social and economic burdens worldwide. A number of studies have shown that interaction between individual genetic vulnerability and environmental risk factors, such as stress, is crucial in psychiatric pathophysiology. In particular, the experience of stressful events in childhood, such as neglect, abuse, or parental loss, was found to increase the risk for development of depression in adult life. Here, to reproduce the gene x environment interaction, we employed an animal model that combines genetic vulnerability with early-life stress. The Flinders Sensitive Line rats (FSL), a validated genetic animal model of depression, and the Flinders Resistant Line (FRL) rats, their controls, were subjected to a standard protocol of maternal separation (MS) from postnatal days 2 to 14. A basal comparison between the two lines for the outcome of the environmental manipulation was performed at postnatal day 73, when the rats were into adulthood. We carried out a global proteomic analysis of purified synaptic terminals (synaptosomes), in order to study a subcellular compartment enriched in proteins involved in synaptic function. Two-dimensional gel electrophoresis (2-DE), mass spectrometry, and bioinformatic analysis were used to analyze proteins and related functional networks that were modulated by genetic susceptibility (FSL vs. FRL) or by exposure to early-life stress (FRL + MS vs. FRL and FSL + MS vs. We found that, at a synaptic level, mainly proteins and molecular pathways related to energy metabolism and cellular remodeling were dysregulated. The present results, in line with previous works, suggest that dysfunction of energy metabolism and cytoskeleton dynamics at a synaptic level could be features of stress-related pathologies, in particular major depression.


Diminished arteriolar responses in nitrate tolerance involve ROS and angiotensin II.
Our purpose was to evaluate hyporesponsivity to nitric oxide (NO)-induced dilation in small arterioles during nitrate tolerance. An Alza osmotic pump was implanted in the left flank of adult rats (n = 56) for continuous administration of nitroglycerin (140 microg/h) or vehicle (propylene glycol). On postoperative day 3, arcade (approximately 50-microm diameter) and terminal (approximately 20 microm) arterioles were observed in the cremaster preparation with in vivo video microscopy. Local vascular responses were obtained with micropipette-applied NO donors, with and without superoxide dismutase (SOD), Mn(III) tetrakis(4-benzoic acid) porphyrin chloride (MnTBAP), or losartan. On day 3, NO-mediated dilation was significantly attenuated in nitroglycerin-treated rats. Attenuation was greater in the terminal arterioles compared with the arcades. Control responses were restored by SOD, MnTBAP, or losartan, suggesting a role for elevated angiotensin II and reactive oxygen species (ROS) as mediators of the attenuated NO dilation (nitrate tolerance). Addition of losartan to the drinking water likewise prevented nitrate tolerance. In summary, terminal arterioles are affected by nitrates to a greater extent than the arcade arterioles that feed them, in a process dependent on angiotensin II and ROS.


Activation of mast cell K+ channels through multiple G protein-linked receptors.
The rat basophilic leukemia (RBL) mast cell line possesses cell surface receptors for adenosine whose ligation markedly potentiates antigen-driven Ca2+ influx and secretion. Here we show that engagement of these receptors and of separate P2 purinergic receptors rapidly activates an outwardly rectifying K+ conductance [GK(OR)] in RBL cells. Activation of GK(OR) by the ligands 5'-(N-ethylcarboxamido)adenosine (NECA), ADP, and ATP was prevented by cytoplasmic guanosine 5'-[beta-thio]diphosphate as well as by pretreatment of the cells with pertussis toxin, implicating mediation by a G protein. Multiple cycles of induction and decay of GK(OR) were produced upon application and removal of ligand. Induction of GK(OR) by either ligand was much faster than the induction caused by guanosine 5'-[gamma-thio]triphosphate (t1/2 < 10 sec vs. 210 sec.). In control cells the maximal whole-cell conductance elicited by ADP (2.25 +/- 0.30 nS) or ATP (2.50 +/- 0.33 nS) was about twice as large as that induced by NECA (1.03 +/- 0.11 nS), and similar to that previously reported for the guanosine 5'-[gamma-thio]triphosphate-elicited GK(OR) in RBL cells (2.58 +/- 1.59 nS). Treatment of RBL cells with dexamethasone upregulated Ca2+ responses to NECA, and it also nearly doubled the maximal conductance elicited by NECA without appreciable effect on responses to ADP or ATP. The failure of water-soluble second messengers to activate GK(OR) and the inability of 11 mM EGTA (< 10 nM Ca2+) to prevent activation by ADP suggest that the relevant pathway is membrane-delimited. Two ion-channel blockers inhibited antigen-stimulated secretion with IC50 values similar to those at which they blocked GK(OR), suggesting that activity of the outwardly rectifying K+ channel may be important for stimulus-response coupling in these cells. Potentiation of the secretory response by NECA may reflect, in part, the activation of GK(OR), which serves to repolarize the membrane more effectively than does the constitutive mechanism, thereby enhancing antigen-driven Ca2+ influx. This channel and its functionally associated receptors may allow neighboring cells of the host to modulate the response of mast cells to exogenous antigen.


Underlying Mechanisms of Allopurinol in Eliminating Renal Toxicity Induced by Melamine-Uric Acid Complex Formation: A Computational Study.
Using molecular dynamics, we address uric acid (UA) replacement by a model small-molecule inhibitor, allopurinol (AP), from its aggregated cluster in a columnar fashion. Experimentally it has been affirmed that AP is efficient in preventing UA-mediated renal stone formation. However, no study has presented the underlying mechanisms yet. Hence, a theoretical approach is presented for mapping the AP, which binds to melamine (MM) and UA clusters. In AP's presence, the higher-order cluster of UA molecules turns into a lower-order cluster, which "drags" fewer MM to them. Consequently, the MM-UA composite structure gets reduced. It is worth noting that UA-AP and AP-MM hydrogen-bonding interactions often play an essential role in reducing the UA-MM cluster size. Interestingly, an AP around UA makes a pillar-like structure, confirmed by defining the point-plane distribution function. The decomposition of the preferential interaction by Kirkwood-Buff integral into different angles like 0°-30°, 30°-60°, and 60°-90° firmly establishes the phenomenon mentioned above. However, the structural order for such π-stacking interactions between AP and UA molecules is not hierarchical but rather more spontaneous. The driving force behind UA-AP-MM composite formation is the favorable complexation energy that can be inferred by computing pairwise binding free energies for all possible combinations. Performing enhanced sampling and quantum calculations further confirms the evidence for UA degradation.


Effect of HSP90 inhibitor in pheochromocytoma PC12 cells: an experimental investigation.
This study aims to investigate whether the expression of heat shock protein 90 (HSP90) is associated with the malignant pheochromocytoma (PHEO) and the effects of 17-allylamino-17-demethoxygeldanamcyin (17-AAG) on the expression of vascular endothelial growth factor (VEGF) in PHEO cell line PC12. The expression of HSP90 was investigated in 38 paraffin-embedded samples of PHEO patients using immunohistochemistry. The time and concentration effects of 17-AAG were investigated in PHEO PC12 cells. Cell proliferation was measured by MTT assay and cell counting. Apoptosis was detected by flow cytometry. Positive staining for HSP90 was found in 14 of 17 malignant (82.35%) and in 5 of 21 (23.81%) benign PHEOs. There existed a significant statistical difference between the malignant group and the benign ones (P < 0.001). 17-AAG inhibited the proliferation of HCC cells in a time- and concentration-dependent manner. The apoptosis rates of PC12 cells after treatment with 0.1 μmol/L for 6, 12, 24, and 48 h were significantly higher than that in blank control group. 17-AAG significantly downregulated VEGF-165 protein level in PC12 cells. This study has confirmed that the specific HSP90 inhibitor 17-AAG can play a therapeutic role in malignant PHEO treatment, and HSP90 qualifies as a promising new target in malignant PHEO.


Morphological Markers for Cryopreservation in the Embryonic Development of Drosophila suzukii (Diptera: Drosophilidae).
Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) is an invasive pest recently reported in Europe whose spread into new areas has caused severe economic damage to many agricultural crops. There are serious concerns about the currently available chemical insecticides because of their low efficacy in controlling the species and their environmental impact; so, several studies have focused on environmentally safe strategies. The sterile insect technique (SIT), which requires colony maintenance in laboratory and production of large numbers of live animals, can be utilized in pest management programs and could be integrated with other control strategies if the potential risks associated with the rearing and maintenance of the insect line under laboratory conditions are given sufficient attention. In this regard, the ability to cryobiologically preserve such stocks would be of substantial value. Important prerequisites for long-term cryopreservation are determination of the embryonic stages, identification of specific embryonic stages, and knowledge of development time. This paper describes the main visible markers for the different stages of embryonic development and determines the timing of development at 25°C. D. suzukii embryogenesis lasts 23-25 h at 25°C and can be divided into 17 stages defined by specific morphological markers. The point at which 50% of embryos are at Stage 14 and 50% are at Stage 15, the most tolerant stages for cryopreservation treatment, as ascertained for Drosophila melanogaster Meigen in prior studies, is reached in 14-15 h. The efficiency of this procedure might be impaired by the retention of eggs in the oviducts, making it impossible to determine the stage of embryonic development for ∼25% of laid eggs.


Frequency of Calreticulin (CALR) Mutation and Its Clinical Prognostic Significance in Essential Thrombocythemia and Primary Myelofibrosis: A Meta-analysis.
Objective As the calreticulin (CALR) mutation frequency is significantly associated with essential thrombocythemia (ET) and primary myelofibrosis (PMF), this mutation may be an important biomarker in patients with ET and PMF. Methods We performed a literature search until April 2015 and obtained 21 relevant studies. The outcome was pooled as the effect size by using the Stata software program. Results The CALR mutation frequencies in patients with ET and PMF were 19% and 22%, respectively. The CALR mutation ratio in Asian patients with ET was 23% and higher than that in European-American patients (16%). Moreover, the mutation ratio in Asian patients with PMF was lower (21%) than that in European-American patients (23%). A slight trend toward fibrotic transformation was found in ET with CALR mutations, whereas leukemic transformation was not significant in patients with ET or PMF with CALR mutations. Conclusion CALR mutations significantly influence the incident of ET as demonstrated by the meta-analysis.


Association of Intraluminal Thrombus, Hemodynamic Forces, and Abdominal Aortic Aneurysm Expansion Using Longitudinal CT Images.
While hemodynamic forces and intraluminal thrombus (ILT) are believed to play important roles on abdominal aortic aneurysm (AAA), it has been suggested that hemodynamic forces and ILT also interact with each other, making it a complex problem. There is, however, a pressing need to understand relationships among three factors: hemodynamics, ILT accumulation, and AAA expansion for AAA prognosis. Hence this study used longitudinal computer tomography scans from 14 patients and analyzed the relationship between them. Hemodynamic forces, represented by wall shear stress (WSS), were obtained from computational fluid dynamics; ILT accumulation was described by ILT thickness distribution changes between consecutives scans, and ILT accumulation and AAA expansion rates were estimated from changes in ILT and AAA volume. Results showed that, while low WSS was observed at regions where ILT accumulated, the rate at which ILT accumulated occurred at the same rate as the aneurysm expansion. Comparison between AAAs with and without thrombus showed that aneurysm with ILT recorded lower values of WSS and higher values of AAA expansion than those without thrombus. Findings suggest that low WSS may promote ILT accumulation and submit the idea that by increasing WSS levels ILT accumulation may be prevented.


A Transient and Flexible Cation-π Interaction Promotes Hydrolysis of Nucleic Acids in DNA and RNA Nucleases.
Metal-dependent DNA and RNA nucleases are enzymes that cleave nucleic acids with great efficiency and precision. These enzyme-mediated hydrolytic reactions are fundamental for the replication, repair, and storage of genetic information within the cell. Here, extensive classical and quantum-based free-energy molecular simulations show that a cation-π interaction is transiently formed in situ at the metal core of Bacteriophage-λ Exonuclease (Exo-λ), during catalysis. This noncovalent interaction (Lys131-Tyr154) triggers nucleophile activation for nucleotide excision. Then, our simulations also show the oscillatory dynamics and swinging of the newly formed cation-π dyad, whose conformational change may favor proton release from the cationic Lys131 to the bulk solution, thus restoring the precatalytic protonation state in Exo-λ. Altogether, we report on the novel mechanistic character of cation-π interactions for catalysis. Structural and bioinformatic analyses support that flexible orientation and transient formation of mobile cation-π interactions may represent a common catalytic strategy to promote nucleic acid hydrolysis in DNA and RNA nucleases.


Single-Crystal Graphene-Directed van der Waals Epitaxial Resistive Switching.
Graphene has been broadcasted as a promising choice of electrode and substrate for flexible electronics. To be truly useful in this regime, graphene has to prove its capability in ordering the growth of overlayers at an atomic scale, commonly known as epitaxy. Meanwhile, graphene as a diffusion barrier against atoms and ions has been shown in some metal-graphene-dielectric configurations for integrated circuits. Guided by these two points, this work explores a new direction of using graphene as a bifunctional material in an electrochemical metallization memory, where graphene is shown to (i) order the growth of a low-ionicity semiconductor ZnS single-crystalline film and (ii) regulate the ion migration in the resistive switching device made of Cu/ZnS/graphene/Cu structures. The ZnS film is confirmed to be van der Waals epitaxially grown on single-crystal graphene with X-ray structural analysis and Raman spectroscopy. Charge transport studies with controlled kinetic parameters reveal superior ion regulating characteristic of graphene in this ZnS-based resistive switching device. The demonstration of the first graphene-directed epitaxial wide band gap semiconductor resistive switching suggests a possible and promising route toward flexible memristors.


Role of the C-terminal di-leucine motif of 5-HT1A and 5-HT1B serotonin receptors in plasma membrane targeting.
The 5-HT1A and 5-HT1B serotonin receptors exhibit different subcellular localizations in neurons. Evidence has been reported that the C-terminal domain is involved in the somato-dendritic and axonal targeting of 5-HT1AR and 5-HT1BR, respectively. Here we analyzed the consequences of the mutation of a di-leucine motif and palmitoylated cysteines within this domain. Replacement of I414-I415 by a di-alanine in 5-HT1AR led to endoplasmic reticulum (ER) sequestration of the corresponding mutant expressed in cell lines as well as in hippocampal neurons in culture. Furthermore, di-leucine-mutated receptors were unable to bind 5-HT1A agonists and presented a major deficit in their glycosylation state, suggesting that they are misfolded. By contrast, mutation of the di-leucine motif in the C-terminal domain of 5-HT1BR had no major consequence on its subcellular targeting. However, in the case of the 1ActB chimera (substitution of the C-terminal domain of the 5-HT1BR into 5-HT1AR), this mutation was also found to cause sequestration within the ER. Replacement of palmitoylated cysteines by serines had no consequence on either receptor type. These data indicate that the di-leucine motif of the 5-HT1AR and 5-HT1BR tails is implicated in proper folding of these receptors, which is necessary for their ER export.


Use of polymerase chain reaction for early detection and management of hepatitis C virus infection after needlestick injury.
Infection with hepatitis C virus (HCV) is a matter of great concern because of its potentially grave consequences. Instead of relying on the conventional anti-HCV antibody test to detect HCV infection after needlestick incidents, we used the polymerase chain reaction (PCR) to achieve earlier detection, to manage a patient more effectively, and to exclude possible infection more quickly. Fourteen incidents were studied in which the source patients were positive for both the anti-HCV antibody and HCV RNA, and the exposed subjects were negative for anti-HCV antibody at the time of the incidents. In one of the exposed subjects, a nurse, the result of the PCR test for HCV RNA was positive at 2 wk after the needlestick incident; the nurse's viral load was very low (800 copies/ml) and she responded well to immediate medical treatment. She never developed acute hepatitis C; her serum anti-HCV antibody level and alanine aminotransferase (ALT) activity did not become elevated, and results of her PCR test for HCV RNA were negative following treatment. In the other 13 needlestick incidents, the results of PCR tests of the exposed subjects were negative for HCV RNA throughout the study and possible infections were quickly ruled out.


Visual quality after wavefront-guided LASIK for myopia.
This study evaluated the visual quality after wavefront-guided laser in situ keratomileusis (LASIK) for treating myopia. Thirty-two eyes with moderate myopia (-5.78 - -2.17D) and 25 eyes with high myopia (-7.78 - -6.17D) were prospectively reviewed. The contrast sensitivity (CS), glare and the total higher order aberrations (HOA) were measured before and 1 week, 1 month and 2 months after LASIK. The pupil diameter was measured at day- and night-time illumination. The CS and glare at all spatial frequencies were not reduced after wavefront-guided LASIK (p<0.05) and the difference between the moderate and high myopia group was not significant. No significant correlation was found between the amounts of myopia and the postoperative CS (p>0.05). The area under the log contrast sensitivity function (AULCSF) showed no correlation with the total HOA (r2=-0.071, p=0.612, between the daytime AULCSF and the total HOA with a 4 mm entrance pupil, r2=-0.176, p=0.260, between the nighttime AULCSF and the total HOA with a 6 mm entrance pupil). There was no decrease in CS and glare after wavefront-guided LASIK for myopia. In conclusion, wavefront-guided LASIK based on the individual ablation patterns is a good option for refractive surgery to improve the visual quality in both moderate and high myopia cases.


Human CD4+ T lymphocytes recognize a vascular endothelial growth factor receptor-2-derived epitope in association with HLA-DR.
Given the multiple escape mechanisms of tumor cells, immunotherapy targeting tumor-dependent stroma may be an effective cancer treatment strategy. Animal models indicate that inducing immunity to tumor endothelia engenders potent antitumor effects without significant pathology. Recently, the first human tumor endothelial antigen vascular endothelial growth factor receptor-2 (VEGFR-2) recognized by HLA class I-restricted CD8(+) T cells has been characterized. In this study, we sought to investigate specific recognition of this molecule by human CD4(+) T cells. To identify HLA-DR-restricted antigenic peptides on VEGFR-2 recognized by CD4(+) T cells of healthy donors and cancer patients. Nine candidate VEGFR-2 peptides with high binding probability to six common HLA-DRB1 alleles were synthesized using the SYFPEITHI algorithm. One 15-mer peptide (EKRFVPDGNRISWDS), mapping to the 167-181 region of VEGFR-2, stimulated CD4(+) T cells in association with several HLA-DR alleles, including DR4 and DR7. Importantly, the epitope could be naturally processed and presented both by HLA-DR-matched antigen-expressing proliferating endothelial cells and by dendritic cells loaded with the native antigen. Furthermore, circulating VEGFR-2-specific CD4(+) T cells were detected in 4 of 10 healthy donors and 12 of 40 cancer patients even after single-round peptide stimulation in short-term culture. Patient's T cells could recognize antigen-expressing proliferating endothelial cells in a HLA-DR-restricted fashion. These findings indicate an important role for the 167-181 region of VEGFR-2 in the stimulation of CD4(+) T cell responses to VEGFR-2 protein, and may be instrumental both for the development and monitoring of upcoming antitumor vessel vaccines against different cancers based on VEGFR-2 immunogens.


Relationship between therapeutic efficacy of doxorubicin-transferrin conjugate and expression of P-glycoprotein in chronic erythromyeloblastoid leukemia cells sensitive and resistant to doxorubicin.
Conjugation of anti-neoplastic agents with human proteins is a strategy to diminish the toxic side effects of anthracycline antibiotics. We have developed a novel doxorubicin-transferrin (DOX-TRF) conjugate aimed to direct anticancer drugs against therapeutic targets that display altered levels of expression in malignant versus normal cells. Our previous work has shown that the cellular bio-distribution of the conjugate is dependent on a dynamic balance between influx and efflux processes. Here, we set out to investigate whether P-glycoprotein (P-gp) expression may affect DOX-TRF conjugate-induced cellular drug accumulation and cytotoxicity. All experiments were carried out on human erythromyeloblastoid cells exhibiting P-gp over-expression (K562/DOX) and its drug sensitive parental line (K562). MTT cytotoxicity, flow cytometry, fluorescence microscopy and RT-PCR assessments revealed that the investigated conjugate (DOX-TRF) possesses a greater cytotoxic potential than free DOX. Our data suggest that the newly developed DOX-TRF conjugate is a less P-gp dependent substrate than free DOX and, consequently, may be used in a clinical setting to increase treatment efficacy in resistant human tumors.