HLA Class II genotypes associated with early-onset periodontitis: DQB1 molecule primarily confers susceptibility to the disease.
DNA typing was performed on 24 Japanese patients with early-onset periodontitis (EOP) using the PCR-RFLP method to investigate an association of the susceptibility to EOP with the particular HLA class II alleles (HLA-DRB1, -DQA1, and -DQB1). DRB1*1401, DRB1*1501, DQB1*0503, and DQB1*0602 were found more frequently ("susceptible") in the EOP patients than in healthy controls. In contrast, DRB1*0405 and DQB1*0401 were found less frequently ("resistant") in EOP patients. All patients carrying DQB1*0602 had an atypical BamHI site in the intron upstream of the third exon of the DQB1 gene, which in our previous studies appeared to be a susceptible marker for EOP. A comparative analysis of the amino acid sequences of these susceptible and resistant HLA-DRB1 and DQB1 alleles elucidated some differences in antigen-derived peptide binding sites related to the susceptible or resistant alleles. Especially, DQB1*0503 and DQB1*0602 alleles carrying aspartic acid at position 57 and glycine at position 70 are increased significantly in EOP. Since amino acid residues at positions 57 and 70 on the DQB1 molecule are supposed to be involved in antigen binding, amino acid substitutions at these positions may affect the immune responsiveness to the periodontopathic antigen. Our results suggest that the DQB1 molecule plays a crucial role in the pathogenesis of EOP and that the susceptibility to EOP may be determined by the binding ability between the peptide and HLA-DQ antigens.


Recombinant Toxoplasma gondii surface antigen 1 (P30) expressed in Escherichia coli is recognized by human Toxoplasma-specific immunoglobulin M (IgM) and IgG antibodies.
The immunodominant surface antigen of Toxoplasma gondii, surface antigen 1 (SAG1), was expressed in Escherichia coli as a fusion protein containing a majority of the SAG1 protein supplied with six histidyl residues in the N-terminal end. The recombinant protein was purified on a Ni-chelate column and then on a fast-performance liquid chromatography column and was in a nonreduced condition. It was recognized by T. gondii-specific human immunoglobulin G (IgG) and IgM antibodies as well as by a mouse monoclonal antibody (S13) recognizing only nonreduced native SAG1. Antibodies induced in mice by the recombinant SAG1 recognized native SAG1 from the T. gondii RH isolate in culture. Recombinant SAG1 is suitable for use in diagnostic systems for detecting anti-SAG1-specific IgG and IgM antibodies.


Phantom limb pain as a manifestation of paclitaxel neurotoxicity.
Paclitaxel is a chemotherapeutic agent with activity directed against several malignancies. It has multiple adverse effects including neurotoxicity. We describe 2 patients with prior amputation who experienced phantom limb pain (PLP) after receiving paclitaxel therapy. A third patient experienced disabling neurotoxicity in the extremity of a prior ulnar nerve and tendon transposition after receiving paclitaxel. This unique syndrome should be identified as a direct causal effect of paclitaxel. In this report, we review the pathophysiology of PLP and treatment options. Physicians should be aware that PLP can occur after initiation of paclitaxel.


Inactivation kinetics of dihydrofolate reductase from Chinese hamster during urea denaturation.
The kinetic theory of substrate reaction during modification of enzyme activity has been applied to the study of inactivation kinetics of Chinese hamster dihydrofolate reductase by urea [Tsou (1988) Adv. Enzymol. Relat. Areas Mol. Biol. 61, 381-436]. On the basis of the kinetic equation of substrate reaction in the presence of urea, all microscopic kinetic constants for the free enzyme and enzyme-substrate binary and ternary complexes have been determined. The results of the present study indicate that the denaturation of dihydrofolate reductase by urea follows single-phase kinetics, and changes in enzyme activity and tertiary structure proceed simultaneously in the unfolding process. Both substrates, NADPH and 7,8-dihydrofolate, protect dihydrofolate reductase against inactivation, and enzyme-substrate complexes lose their activity less rapidly than the free enzyme.


Increased hepatic activity of inducible nitric oxide synthase in rats fed on a high-fat diet.
The effects were examined of the dietary level of fat on the activity of inducible nitric oxide synthase (iNOS) in the liver of rats. In experiment 1, rats were fed on a diet containing 5% or 20% beef tallow or safflower oil for 32 d. The animals were given a subcutaneous injection of the carcinogen, 1,2-dimethylhydrazine (DMH), on d 4. The activity of hepatic iNOS was significantly elevated by the high-fat diet, but was unaffected by the dietary source of the fat examined. In experiment 2, rats were fed on a 5% or 20% beef tallow diet for 11 d or 32 d with or without the DMH treatment. Feeding the high-fat diet and DMH treatment caused higher activity of hepatic iNOS. In experiment 3, the high-fat diet elevated hepatic iNOS activity and the amount of its protein in the lipopolysaccharide-treated rats. The results suggest that hepatic NO production is enhanced by a high-fat diet.


Synonymous genome recoding: a tool to explore microbial biology and new therapeutic strategies.
Synthetic genome recoding is a new means of generating designed organisms with altered phenotypes. Synonymous mutations introduced into the protein coding region tolerate modifications in DNA or mRNA without modifying the encoded proteins. Synonymous genome-wide recoding has allowed the synthetic generation of different small-genome viruses with modified phenotypes and biological properties. Recently, a decreased cost of chemically synthesizing DNA and improved methods for assembling DNA fragments (e.g. lambda red recombination and CRISPR-based editing) have enabled the construction of an Escherichia coli variant with a 4-Mb synthetic synonymously recoded genome with a reduced number of sense codons (n = 59) encoding the 20 canonical amino acids. Synonymous genome recoding is increasing our knowledge of microbial interactions with innate immune responses, identifying functional genome structures, and strategically ameliorating cis-inhibitory signaling sequences related to splicing, replication (in eukaryotes), and complex microbe functions, unraveling the relevance of codon usage for the temporal regulation of gene expression and the microbe mutant spectrum and adaptability. New biotechnological and therapeutic applications of this methodology can easily be envisaged. In this review, we discuss how synonymous genome recoding may impact our knowledge of microbial biology and the development of new and better therapeutic methodologies.


Effects of Graded Exergames on Fitness Performance in Elementary School Children With Developmental Coordination Disorder.
Developmental Coordination Disorder (DCD) is a common childhood disorder affecting movement and coordination skills, fitness, and academic performance. Increased physical fitness may have a positive influence on physical and mental health outcomes in children with DCD. Yet, little has been done to develop interventions to improve fitness performance in this group. The purpose of this study was to determine the effects of graded exergames in 7 to 12-year-old children with DCD and typically developing (TD) peers. Participants (32 DCD and 28 TD children) received a 30-min training session twice weekly for 10 weeks. Performance on motor coordination (MABC-2 test), balance, aerobic, and anaerobic fitness tests were assessed at the beginning and end of training. In addition, enjoyment and perception of exertion were measured for each participant during the training period. Both children with DCD and TD children significantly improved on motor coordination, balance, aerobic, and anaerobic fitness at the end of the training. A significant Group by Time interaction was observed on the MABC-2 total [F (1, 55) = 13.19; p < 0.001] and balance scores [F (1, 55) = 26.83; p < 0.0001], with the DCD group demonstrating larger improvements than the TD children. Both groups enjoyed the program throughout the training period even though they rated the training to be of high intensity. These findings indicate that graded exergames may serve as potential treatment for impaired fitness in children with DCD. Regular participation in graded exergames in school settings may be needed to enhance and maintain fitness performance in young children with and without DCD.


Two types of block copolymer micelles with ion-containing cores.
We report a combined experimental and theoretical study of micellization of block copolymer with hydrophilic nonionic corona-forming blocks and weak polyelectrolyte (wPE) core-forming blocks with pH-triggered solubility in aqueous solutions. We demonstrate that in addition to micelles with neutral cores, there exist two other types of micelles with PE- or ionomer-like cores, in which monovalent counterions are released or condensed on core wPE block, respectively. The transition between the two types of micelles occurred upon changes in ionization of the PE core block and resulted in nonmonotonous changes of aggregation number as a function of pH. Such micelles with stimulus responsive cores represent promising nanocarriers for controlled delivery applications.


Frailty Before Critical Illness and Mortality for Elderly Medicare Beneficiaries.
To estimate the effect of pre-intensive care unit (ICU) health categories on mortality during and after critical illness, focusing specifically on the effect of pre-ICU frailty on short- and long-term mortality. Retrospective cohort study. Medicare claims data from 2004 to 2008. A nationally representative sample of elderly Medicare beneficiaries admitted to an ICU in 2005. Participants were classified into four pre-ICU health categories (robust, cancer, chronic organ failure, frailty) using claims data from the year before admission, allowing for assignment to multiple categories. The association between pre-ICU health category and hospital and 3-year mortality was assessed using multivariable logistic regression and Cox proportional hazards models. Of 47,427 elderly individuals in the ICU, 18.8% were robust, 28.6% had cancer, 68.1% had chronic organ failure, and 34.0% were frail; 41.3% qualified for multiple categories. Overall hospital mortality was 12.6%, with the lowest mortality for robust participants (9.7%). Participants with pre-ICU frailty had higher hospital mortality than those with the same pre-ICU health categories without frailty. (Adjusted odds ratios ranged from 1.27 (95% confidence interval (CI) 1.10-1.47) to 1.52 (95% CI = 1.35-1.63).) Robust hospital survivors had the lowest 3-year mortality (24.6%). Pre-ICU frailty conferred higher 3-year mortality than pre-ICU categories without frailty. (Adjusted hazard ratios ranged from 1.54 (95% CI = 1.45-1.64) to 1.84 (95% CI = 1.70-1.99).) Critically ill elderly adults can be categorized according to pre-ICU health categories. These categories, particularly pre-ICU frailty, may be important for understanding risk of death during and after critical illness.


Mechanisms of flagellar excision. I. The role of intracellular acidification.
It has previously been shown that exposure of Chlamydomonas to low pH induces the cells to shed their flagella. In the present paper we report that a 30-s treatment with 20 mM acetic, carbonic, formic, or benzoic acid at pH 4.0 will induce flagellar excision. In contrast, 20 mM concentrations of the stronger aspartic, phosphoric, citric, and tartaric acids (pH 4) do not induce excision. Further, the excision efficacy of acetate is a function of the concentration of protonated acetate. Thus, excision correlates with the presence of a protonated, membrane-permeant species of acid. Relative to acetate, the more permeant benzoate induces excision at a much lower concentration of protonated acid. We conclude that a flux of acid into the cell is the signal for excision. Previous work has shown that detergent-permeabilized cells excise their flagella in response to calcium but not in response to low pH. This suggests that the acidification of intact cells triggers excision by stimulating an increase in intracellular calcium. We have previously reported that the source of this calcium might be IP3-sensitive. In our model for the mechanism of pH-induced flagellar excision, a flux of acid into the cell activates phospholipase C, leading to IP3 production, the activation of an IP3-gated calcium channel (located on either an intracellular or surface membrane), and an increase in cytosolic calcium, which is the trigger of flagellar excision.


Lung cancer surgery: an up to date.
According to the International Agency for Research on Cancer (IARC) GLOBOCAN World Cancer Report, lung cancer affects more than 1 million people a year worldwide. In Greece according to the 2008 GLOBOCAN report, there were 6,667 cases recorded, 18% of the total incidence of all cancers in the population. Furthermore, there were 6,402 deaths due to lung cancer, 23.5% of all deaths due to cancer. Therefore, in our country, lung cancer is the most common and deadly form of cancer for the male population. The most important prognostic indicator in lung cancer is the extent of disease. The Union Internationale Contre le Cancer (UICC) and the American Joint Committee for Cancer Staging (AJCC) developed the tumour, node, and metastases (TNM) staging system which attempts to define those patients who might be suitable for radical surgery or radical radiotherapy, from the majority, who will only be suitable for palliative measures. Surgery has an important part for the therapy of patients with lung cancer. "Lobectomy is the gold standard treatment". This statement may be challenged in cases of stage Ia cancer or in patients with limited pulmonary function. In these cases an anatomical segmentectomy with lymph node dissection is an acceptable alternative. Chest wall invasion is not a contraindication to resection. En-bloc rib resection and reconstruction is the treatment of choice. N2 disease represents both a spectrum of disease and the interface between surgical and non-surgical treatment of lung cancer Evidence from trials suggests that multizone or unresectable N2 disease should be treated primarily by chemoradiotherapy. There may be a role for surgery if N2 is downstaged to N0 and lobectomy is possible, but pneumonectomy is avoidable. Small cell lung cancer (SCLC) is considered a systemic disease at diagnosis, because the potential for hematogenous and lymphogenic metastases is very high. The efficacy of surgical intervention for SCLC is not clear. Lung cancer resection can be performed using several surgical techniques. Video-assisted thoracoscopic surgery (VATS) lobectomy is a safe, efficient, well accepted and widespread technique among thoracic surgeons. The 5-year survival rate following complete resection of lung cancer is stage dependent. Incomplete resection rarely is useful and cures the patient.


Reaction dynamics through kinetic transition states.
The transformation of a system from one state to another is often mediated by a bottleneck in the system's phase space. In chemistry, these bottlenecks are known as transition states through which the system has to pass in order to evolve from reactants to products. The chemical reactions are usually associated with configurational changes where the reactants and products states correspond, e.g., to two different isomers or the undissociated and dissociated state of a molecule or cluster. In this Letter, we report on a new type of bottleneck which mediates kinetic rather than configurational changes. The phase space structures associated with such kinetic transition states and their dynamical implications are discussed for the rotational vibrational motion of a triatomic molecule. An outline of more general related phase space structures with important dynamical implications is given.


Role of Bisphosphonates in Postmenopausal Women with Osteoporosis to Prevent Future Fractures: A Literature Review.
Postmenopausal women who have osteoporosis are at increased risk of future fractures. Bisphosphonates are drugs that are used to treat osteoporosis by acting on the osteoclasts to inhibit bone resorption. Several studies have shown that bisphosphonates can maintain or even increase bone mineral density in osteoporosis patients. This review study analyzed the literature on clinical experiments with bisphosphonate therapy in postmenopausal women to determine if these drugs are efficacious in preventing future fractures. Four out of five studies found that women treated with bisphosphonates were at a decreased risk of future fractures, and six of six studies found that bisphosphonate therapy increases bone mineral density relative to placebo control. Although further work is warranted to understand the level of bone mineral density increase that is associated with fracture prevention, this study implies that bisphosphonate therapy can be used to help prevent future fractures in postmenopausal osteoporotic women. The study is significant in that it helps to underscore the efficacy of bisphosphonate therapy in postmenopausal women, and it may be generalizable to other populations with osteoporosis who are at increased risk of fractures.


Multiple behavioural, morphological and cognitive developmental changes arise from a single alteration to early life spatial environment, resulting in fitness consequences for released pheasants.
Subtle variations in early rearing environment influence morphological, cognitive and behavioural processes that together impact on adult fitness. We manipulated habitat complexity experienced by young pheasants (Phasianus colchicus) in their first seven weeks, adding a third accessible dimension by placing elevated perches in their rearing pens mimicking natural variation in habitat complexity. This simple manipulation provoked an interrelated suite of morphological, cognitive and behavioural changes, culminating in decreased wild mortality of birds from complex habitats compared with controls. Three mechanisms contribute to this: Pheasants reared with perches had a morphology which could enable them to fly to the higher branches and cope with prolonged roosting. They had a higher propensity to roost off the ground at night in the wild. More generally, these birds had more accurate spatial memory. Consequently, birds were at a reduced risk of terrestrial predation. The fitness consequences of variation in early rearing on behavioural development are rarely studied in the wild but we show that this is necessary because the effects can be broad ranging and not simple, depending on a complex interplay of behavioural, cognitive and morphological elements, even when effects that the treatments provoke are relatively short term and plastic.


CausalBuilder: bringing the MI2CAST causal interaction annotation standard to the curator.
Molecular causal interactions are defined as regulatory connections between biological components. They are commonly retrieved from biological experiments and can be used for connecting biological molecules together to enable the building of regulatory computational models that represent biological systems. However, including a molecular causal interaction in a model requires assessing its relevance to that model, based on the detailed knowledge about the biomolecules, interaction type and biological context. In order to standardize the representation of this knowledge in 'causal statements', we recently developed the Minimum Information about a Molecular Interaction Causal Statement (MI2CAST) guidelines. Here, we introduce causalBuilder: an intuitive web-based curation interface for the annotation of molecular causal interactions that comply with the MI2CAST standard. The causalBuilder prototype essentially embeds the MI2CAST curation guidelines in its interface and makes its rules easy to follow by a curator. In addition, causalBuilder serves as an original application of the Visual Syntax Method general-purpose curation technology and provides both curators and tool developers with an interface that can be fully configured to allow focusing on selected MI2CAST concepts to annotate. After the information is entered, the causalBuilder prototype produces genuine causal statements that can be exported in different formats.


Preventing problems in Ecstasy users: reduce use to reduce harm.
Increasing use of 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy") has been accompanied by concern about acute and possible long-term toxicity. This article discusses acute serious toxicity, chronic toxicity, and common problems associated with Ecstasy use, as well as the implications of these areas for prevention programs targeted at current Ecstasy users. The low incidence of serious adverse events in users creates difficulties for attempts to develop harm reduction recommendations. Many hypothesized risk factors for serious adverse events cannot be confirmed or denied and may not be associated with dramatic elevations in risk. Research on chronic toxicity in users provides strong evidence of neurophysiological changes and suggestive evidence of possible neurocognitive changes. Because these worrisome changes are clinically subtle, users may not be influenced by concerns of neurotoxicity. In contrast, common Ecstasy-related complaints are relatively well documented and have identified risk factors, including factors relating to extent of Ecstasy use (such as "binges"). Common complaints include modest acute and subacute adverse effects,some lasting several days, and problems in life. The apparent willingness of users to modify drug use and other behaviors to decrease these common problems could be used by harm reduction or other prevention programs to encourage users to decrease the extent of Ecstasy use.


Autologous T Cells Expressing CD30 Chimeric Antigen Receptors for Relapsed or Refractory Hodgkin Lymphoma: An Open-Label Phase I Trial.
Purpose: Relapsed or refractory Hodgkin lymphoma is a challenge for medical oncologists because of poor overall survival. We aimed to assess the feasibility, safety, and efficacy of CD30-targeting CAR T cells in patients with progressive relapsed or refractory Hodgkin lymphoma.Experimental Design: Patients with relapsed or refractory Hodgkin lymphoma received a conditioning chemotherapy followed by the CART-30 cell infusion. The level of CAR transgenes in peripheral blood and biopsied tumor tissues was measured periodically according to an assigned protocol by quantitative PCR (qPCR).Results: Eighteen patients were enrolled; most of whom had a heavy treatment history or multiple tumor lesions and received a mean of 1.56 × 107 CAR-positive T cell per kg (SD, 0.25; range, 1.1-2.1) in total during infusion. CART-30 cell infusion was tolerated, with grade ≥3 toxicities occurring only in two of 18 patients. Of 18 patients, seven achieved partial remission and six achieved stable disease. An inconsistent response of lymphoma was observed: lymph nodes presented a better response than extranodal lesions and the response of lung lesions seemed to be relatively poor. Lymphocyte recovery accompanied by an increase of circulating CAR T cells (peaking between 3 and 9 days after infusion) is a probable indictor of clinical response. Analysis of biopsied tissues by qPCR and immunohistochemistry revealed the trafficking of CAR T cells into the targeted sites and reduction of the expression of CD30 in tumors.Conclusions: CART-30 cell therapy was safe, feasible, and efficient in relapsed or refractory lymphoma and guarantees a large-scale patient recruitment. Clin Cancer Res; 23(5); 1156-66. ©2016 AACR.


Penile Sarcoma: Report of a Rare Malignancy.
Penile cancer is an uncommon malignancy. Squamous cell carcinoma constitutes approximately 95% of all histology. Non-squamous malignancies are rare in penis. Sarcomas of penis are rarer among them. Spindle cell sarcoma is one of the extremely rare sarcoma of penis. To best of our knowledge, only two cases have been reported so far, one in English literature and other in Japanese. We are presenting this uncommon case of spindle cell sarcoma of penis, which was diagnosed with microscopy with its characteristic immunohistochemistry. The disease had an aggressive course with multiple recurrences in a short duration despite margin negative resection. Disease responded poorly with the chemotherapy and patient succumbed to the disease.


D4F104S1 deletion in facioscapulohumeral muscular dystrophy: phenotype, size, and detection.
The facioscapulohumeral muscular dystrophy (FSHD) locus maps to 4q35 where it is closely linked to D4F104S1 (p13E-11), a probe that recognizes the pathognomonic FSHD deletion involving the subtelomeric D4Z4 tandem repeat array. Extended deletions that include both the more proximal D4F104S1 region and the D4Z4 repeat array proper do, however, occur, albeit rarely, and such deletions can lead to difficulties of interpretation in the diagnostic setting. To devise a means to determine the true frequency of proximally extended deletions in individuals with FSHD. Three families selected for this study were originally identified during routine FSHD analysis on the basis that the affected individuals in each family had failed to exhibit a small (<38-kb) EcoRI fragment. High molecular weight DNA from these families was analyzed with both conventional and pulsed-field gel electrophoresis using DNA markers p13E-11, 9B6A, B31, 4qA, and 4qB. Large genomic deletions were identified involving both D4Z4 and D4F104S1. The precise number of D4Z4 repeat units borne by the p13E11 deletion allele was established by the use of an additional restriction enzyme (MseI) digest. All three cases carry different sizes of deletion proximal to the D4Z4 repeat units. With use of a recently described telomeric probe, 4qA, a method was developed that identifies large genomic deletions involving both D4Z4 and D4F104S1 using conventional gel electrophoresis. Proximally extended deletions can be found in patients with a normal spectrum of the disease. This assay promises to allow estimation of the true frequency of proximally extended deletions and should improve the accuracy and reliability of molecular diagnostic testing for FSHD.


Penicillin susceptibility breakpoints for Streptococcus pneumoniae and their effect on susceptibility categorisation in Germany (1997-2013).
Continuous nationwide surveillance of invasive pneumococcal disease (IPD) was conducted in Germany. From July 1, 1997, to June 30, 2013, data on penicillin susceptibility were available for 20,437 isolates. 2,790 of these isolates (13.7 %) originate from patients with meningitis and 17,647 isolates (86.3 %) are from non-meningitis cases. A slight decline in isolates susceptible at 0.06 and 0.12 μg/ml can be noticed over the years. Overall, 89.1 % of the isolates had minimum inhibitory concentrations (MICs) of ≤0.015 μg/ml. In 2012/2013, the first three isolates of Streptococcus pneumoniae with MICs of 8 μg/ml were found. The application of different guidelines with other MIC breakpoints for the interpretation of penicillin resistance leads to differences in susceptibility categorisation. According to the pre-2008 Clinical and Laboratory Standards Institute (CLSI) interpretive criteria, 5.3 % of isolates overall were intermediate and 1.4 % were resistant to penicillin. Application of the 2008-2014 CLSI interpretive criteria resulted in 7.6 % resistance among meningitis cases and 0.5 % intermediate resistance in non-meningitis cases. Referring to the 2009-2014 European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, 7.6 % of the isolates in the meningitis group were resistant to penicillin. In the non-meningitis group, 6.1 % of the isolates were intermediate and 0.5 % were resistant. These differences should be kept in mind when surveillance studies on pneumococcal penicillin resistance are compared.


Isolate-Dependent Differences in Clinical, Pathological, and Transcriptional Profiles following In Vitro and In Vivo Infections with Rickettsia rickettsii.
Rickettsia rickettsii, the etiological agent of Rocky Mountain spotted fever (RMSF), a life-threatening tick-borne disease that affects humans and various animal species, has been recognized in medicine and science for more than 100 years. Isolate-dependent differences in virulence of R. rickettsii have been documented for many decades; nonetheless, the specific genetic and phenotypic factors responsible for these differences have not been characterized. Using in vivo and in vitro methods, we identified multiple phenotypic differences among six geographically distinct isolates of R. rickettsii, representing isolates from the United States, Costa Rica, and Brazil. Aggregate phenotypic data, derived from growth in Vero E6 cells and from clinical and pathological characteristics following infection of male guinea pigs (Cavia porcellus), allowed separation of these isolates into three categories: nonvirulent (Iowa), mildly virulent (Sawtooth and Gila), and highly virulent (Sheila SmithT, Costa Rica, and Taiaçu). Transcriptional profiles of 11 recognized or putative virulence factors confirmed the isolate-dependent differences between mildly and highly virulent isolates. These data corroborate previous qualitative assessments of strain virulence and suggest further that a critical and previously underappreciated balance between bacterial growth and host immune response could leverage strain pathogenicity. Also, this work provides insight into isolate-specific microbiological factors that contribute to the outcome of RMSF and confirms the hypothesis that distinct rickettsial isolates also differ phenotypically, which could influence the severity of disease in vertebrate hosts.


Outcome after surgical treatment for lumbar spinal stenosis: the lumbar extension test is not a predictive factor.
A prospective clinical study. To investigate the predictive value of the lumbar extension test for outcome after surgical treatment of lumbar spinal stenosis (LSS). Studies have indicated that aggravation of the symptoms from LSS by extension of the lumbar spine has predictive value for the outcome after decompression. The aim of this study was to investigate this theory in a larger group of patients. One hundred forty-six consecutive patients surgically treated for LSS were included in the study. The clinical condition was recorded before surgery and at 3, 6, 12, and 24 months after surgery using 3 different scoring systems: Swiss Spinal Stenosis Questionnaire, Neurogenic Claudication Outcome Score, and Oswestry Disability Index. The group of patients with preoperative aggravation of the symptoms by the lumbar extension test, (positive extension test), was compared with the group of patients without aggravation by the test, (negative extension test). Before surgery, patients with a positive extension test scored significantly worse on all disability scoring systems than patients with a negative test. However, the extension test itself had no prognostic value for the overall outcome after lumbar decompression. Using regression models with the 2-year Oswestry Disability Index as dependent variable, only before surgery self-reported health and age were found to have prognostic significance. The lumbar extension test has no predictive value for the outcome after surgical treatment of LSS.


Isomeric fatty acids in the US diet: levels and health perspectives.
Data on market size, market share, and compositional changes of various products made from partially hydrogenated fats and oils were used to estimate current, previous, and predicted levels of trans fatty acids in the US diet. Recent literature and various perceptions of the toxicological, physiological, and nutritional effects of trans fatty acids were reviewed. Trans acid availability has changed little during the last 15-20 yr and currently is estimated to be approximately 7.6 g X person-1 X day-1. Modest decreases in total fat intake, including trans fatty acid intake, may occur in the future if consumers adopt certain dietary recommendations. We believe that concerns about possible relationships of trans fatty acids to development of atherosclerotic disease or cancer are not supported by reliable data. Numerous literature accounts support the conclusion that trans fatty acids do not pose any harm to humans or animals consuming a balanced diet containing adequate linoleic acid.


Does erythropoietin protect the preterm brain?
There is a high incidence of hypoxic-ischaemic brain injury and intraventricular haemorrhage in newborn infants, particularly those born preterm. Many die during the newborn period or suffer permanent neurodevelopmental handicaps. Hypoxic brain injury develops over several hours and could potentially be influenced by intervention. At present, no drug exists that effectively prevents infant brain injury or ameliorates detrimental neurodevelopmental effects. The hypothesis is put forward that systemic administration of recombinant human erythropoietin positively affects the neurodevelopmental outcome of high risk preterm infants affected by brain injury. A multicentre, randomised, placebo controlled study is proposed to prospectively test this hypothesis.


[Evaluation of the cardiorespiratory system during rehabilitation of patients after prosthesis of the mitral and annuloplasty of the tricuspid valves].
Twenty-five patients undergoing rehabilitation for 2 to 3 years after surgery were examined. External respiratory parameters were investigated by means of bicycle spiroergometry, and the pump and contractile functions of the heart were assessed using tetrapolar chest rheography. In addition to positive clinical developments and improved functional class, physical working capacity of the patients was found to be increased. External respiratory function was improved accordingly. The efficacy of surgical correction of the defect was evidenced early after surgery by the normalization of minute flow volume and only slightly reduced stroke volume at rest. Hemodynamic support of physical working capacity that increases progressively in the course of rehabilitation is characterized by overstrain of inotropic mechanisms, uneconomically increased minute volume, and insufficiently increased stroke volume, mostly due to the incompetence of the Frank-Starling mechanism associated with tachycardia that is inadequate to the effort.


Inference for a Large Directed Acyclic Graph with Unspecified Interventions.
Statistical inference of directed relations given some unspecified interventions (i.e., the intervention targets are unknown) is challenging. In this article, we test hypothesized directed relations with unspecified interventions. First, we derive conditions to yield an identifiable model. Unlike classical inference, testing directed relations requires to identify the ancestors and relevant interventions of hypothesis-specific primary variables. To this end, we propose a peeling algorithm based on nodewise regressions to establish a topological order of primary variables. Moreover, we prove that the peeling algorithm yields a consistent estimator in low-order polynomial time. Second, we propose a likelihood ratio test integrated with a data perturbation scheme to account for the uncertainty of identifying ancestors and interventions. Also, we show that the distribution of a data perturbation test statistic converges to the target distribution. Numerical examples demonstrate the utility and effectiveness of the proposed methods, including an application to infer gene regulatory networks. The R implementation is available at https://github.com/chunlinli/intdag.


An energizing role for motivation in information-seeking during the early phase of the COVID-19 pandemic.
The COVID-19 pandemic has highlighted the importance of understanding and managing information seeking behavior. Information-seeking in humans is often viewed as irrational rather than utility maximizing. Here, we hypothesized that this apparent disconnect between utility and information-seeking is due to a latent third variable, motivation. We quantified information-seeking, learning, and COVID-19-related concern (which we used as a proxy for motivation regarding COVID-19 and the changes in circumstance it caused) in a US-based sample (n = 5376) during spring 2020. We found that self-reported levels of COVID-19 concern were associated with directed seeking of COVID-19-related content and better memory for such information. Interestingly, this specific motivational state was also associated with a general enhancement of information-seeking for content unrelated to COVID-19. These effects were associated with commensurate changes to utility expectations and were dissociable from the influence of non-specific anxiety. Thus, motivation both directs and energizes epistemic behavior, linking together utility and curiosity.


Exploring the Prospects of Engineered Newcastle Disease Virus in Modern Vaccinology.
Many traditional vaccines have proven to be incapable of controlling newly emerging infectious diseases. They have also achieved limited success in the fight against a variety of human cancers. Thus, innovative vaccine strategies are highly needed to overcome the global burden of these diseases. Advances in molecular biology and reverse genetics have completely restructured the concept of vaccinology, leading to the emergence of state-of-the-art technologies for vaccine design, development and delivery. Among these modern vaccine technologies are the recombinant viral vectored vaccines, which are known for their incredible specificity in antigen delivery as well as the induction of robust immune responses in the vaccinated hosts. Although a number of viruses have been used as vaccine vectors, genetically engineered Newcastle disease virus (NDV) possesses some useful attributes that make it a preferable candidate for vectoring vaccine antigens. Here, we review the molecular biology of NDV and discuss the reverse genetics approaches used to engineer the virus into an efficient vaccine vector. We then discuss the prospects of the engineered virus as an efficient vehicle of vaccines against cancer and several infectious diseases of man and animals.


Selective phosphodiesterase inhibitors modulate the activity of alveolar macrophages from sensitized guinea-pigs.
The aim of this study was to investigate the effects of selective phosphodiesterase (PDE)3 and PDE4 inhibitors on arachidonate release by alveolar macrophages from sensitized and challenged guinea-pigs. Guinea-pigs were sensitized and challenged with ovalbumin administered by aerosol. Bronchoalveolar lavage was performed 48 h later and the PDE and cyclic adenosine monophosphate (cAMP) contents of or the arachidonate release from alveolar macrophages, stimulated in vitro with N-formyl-Met-Leu-Phe (fMLP), were evaluated. PDE3 and PDE4 activities were detected in preparations of macrophage lysate from sensitized challenged and sensitized control animals. Oral pretreatment, prior to antigen challenge in sensitized guinea-pigs, with rolipram or Ro 20-1724 (PDE4 inhibitors) but not milrinone (PDE3 inhibitor) significantly reduced the arachidonate release from alveolar macrophages. In vitro incubation of alveolar macrophages from challenged guinea-pigs with Ro 20-1724 or the cAMP analogue dibutyryl cAMP (db-cAMP) but not milrinone or the cyclic guanosine monophosphate (cGMP) analogue 8-bromo-cGMP (8-br-cGMP) significantly reduced arachidonate release. Incubation of the cells with a combination of milrinone plus rolipram or Ro 20-1724 elicited a marked and significant reduction in arachidonate release by alveolar macrophages stimulated with fMLP. In conclusion, these data show that phosphodiesterase-4 isoenzyme may regulate the release of inflammatory mediators such as arachidonate from macrophages through an increase in intracellular cyclic adenosine monophosphate. This suggests that phosphodiesterase-4 inhibitors have potential in the treatment of inflammatory disorders of the lung.


Influence of bolus size and chewing side on temporomandibular joint intra-articular space during mastication.
Previous studies suggested that, during mastication, magnitude and location of mechanical load in the temporomandibular joint (TMJ) might depend on chewing side and bolus size. Aim of this study was to dynamically measure the TMJ space while chewing on standardized boluses to assess the relationship among minimum intra-articular distances (MID), their location on the condylar surface, bolus size, and chewing side. Mandibular movements of 12 participants (6f, 24±1y.o.; 6 m, 28±6y.o.) were tracked optoelectronically while chewing unilaterally on rubber boluses of 15 × 15 × 5, 15 × 15 × 10, and 15 × 15 × 15 mm3 size. MID and their location along the main condylar axis were determined with dynamic stereometry. MID were normalized on the intra-articular distance in centric occlusion. Repeated measures ANOVA (α = 0.05) showed that MID were smaller on the balancing (0.74±0.19) than on the working condyle (0.89±0.16) independently of bolus size (p < 0.0001). MIDs did not differ between 5 and 10 mm bolus thicknesses (0.80±0.17) but increased for 15 mm (0.85±0.22, p = 0.024) and were located mostly laterally, close to the condylar center. This study confirmed higher reduction of TMJ space on the balancing than on the working condyle during mastication. Intra-articular distances increased significantly for the greatest bolus thickness. Loaded areas were located laterally, for both working and balancing joint.


Depression and Subclinical Coronary Atherosclerosis in Adults Without Clinical Coronary Artery Disease.
The relationship between depression and subclinical coronary atherosclerosis in asymptomatic individuals is not clear. We evaluated this relationship in a Korean population. We analyzed 3920 individuals (mean age 54.7±7.9 years and 2603 men [66.4%]) with no history of coronary artery disease who voluntarily underwent coronary computed tomographic angiography and screening for depression using the Beck Depression Inventory as part of a general health examination. The degree and extent of subclinical coronary atherosclerosis were evaluated by coronary computed tomographic angiography, and ≥50% diameter stenosis was defined as significant. Participants were categorized into groups of those with or without depression using the Beck Depression Inventory scores ≥16 as a cutoff value. Of the study participants, 272 (6.9%) had a Beck Depression Inventory score of 16 or higher. After adjustment for cardiovascular risk factors, depression was not significantly associated with any coronary plaque (adjusted odds ratio [OR], 1.05 [95% CI, 0.78-1.41]; P=0.746), calcified plaque (OR, 0.95 [95% CI, 0.71-1.29]; P=0.758), noncalcified plaque (OR, 1.31 [95% CI, 0.79-2.17]; P=0.305), mixed plaque (OR, 1.16 [95% CI, 0.60-2.23]; P=0.659), or significant coronary artery stenosis (OR, 1.22 [95% CI, 0.73-2.03]; P=0.450). In the propensity score-matched population (n=1318) as well, none of the coronary artery disease measures of subclinical coronary atherosclerosis were statistically significantly associated with depression (all P>0.05). In this large cross-sectional study with asymptomatic individuals undergoing coronary computed tomographic angiography and Beck Depression Inventory evaluation, depression was not associated with an increased risk of subclinical coronary atherosclerosis.


Genotype on the pigmentation regulating PMEL17 gene affects behavior in chickens raised without physical contact with conspecifics.
Chickens homozygous for the Dominant white or wild-type allele of PMEL17 were subjected to a broad phenotyping in order to detect consistent differences between genotypes. To exclude feather pecking, the chickens were individually housed without physical contact, from the day of hatching, and tested for social, aggressive, fear and exploratory behaviors, and corticosterone and testosterone levels were assessed. In a principal component analysis, 53.2% of the behavior variation was explained by two factors. Factor one was an activity and social factor, and there was a significant effect of genotype on the factor scores. On factor two, related to aggressive behavior, there were significant effects of genotype, sex and their interaction. There were no genotype effects on hormone levels or any other measured non-behavioral phenotypes. Hence, differences in behavior between PMEL17 genotypes remained when negative social experiences were excluded, indicating a direct pleiotropic effect of the gene on behavior.


Output feedback control of a quadrotor UAV using neural networks.
In this paper, a new nonlinear controller for a quadrotor unmanned aerial vehicle (UAV) is proposed using neural networks (NNs) and output feedback. The assumption on the availability of UAV dynamics is not always practical, especially in an outdoor environment. Therefore, in this work, an NN is introduced to learn the complete dynamics of the UAV online, including uncertain nonlinear terms like aerodynamic friction and blade flapping. Although a quadrotor UAV is underactuated, a novel NN virtual control input scheme is proposed which allows all six degrees of freedom (DOF) of the UAV to be controlled using only four control inputs. Furthermore, an NN observer is introduced to estimate the translational and angular velocities of the UAV, and an output feedback control law is developed in which only the position and the attitude of the UAV are considered measurable. It is shown using Lyapunov theory that the position, orientation, and velocity tracking errors, the virtual control and observer estimation errors, and the NN weight estimation errors for each NN are all semiglobally uniformly ultimately bounded (SGUUB) in the presence of bounded disturbances and NN functional reconstruction errors while simultaneously relaxing the separation principle. The effectiveness of proposed output feedback control scheme is then demonstrated in the presence of unknown nonlinear dynamics and disturbances, and simulation results are included to demonstrate the theoretical conjecture.


Developmental stability in the mouse vertebral column.
The effects of six mutant genes (Sd, T, vt, Sp, un, Ph) on the axial skeleton of the mouse were studied both alone and in combination with 24 hours starvation on Day 8 of gestation. Normal controls showed a craniocaudal gradient of size variation and relatively low variability towards the boundaries of the cervical, thoracic and lumbar vertebral classes. Gene-induced craniocaudal gradients of abnormality were observed in response to Sd and T, and to Ph, which have direct and indirect effects respectively on the primitive streak and/or notochord. By contrast, vt, which influences the paraxial mesoderm, and Sp and un, which act later, during somite formation and sclerotome differentiation respectively, did not produce craniocaudal gradients of abnormality. For all genes there were indications of relative resistance to their effects towards the vertebral class boundaries. Starvation did not materially influence these patterns of abnormality. The division of the axial skeleton of the mouse into its morphologically distinct classes of vertebrae thus appears to originate at the earliest stage of axial development, even before somite formation, with the positions of the future vertebral class boundaries established both axially and paraxially as regions of relative stability that persist into later stages of development. The primary craniocaudal gradient associated with the advancing primitive streak, from which the vertebral class boundary positions may be derived, appears to be restricted to the axial structures themselves and to be lost after the somites appear.


Longitudinal survey of lymphocyte subpopulations in the first year of life.
Age-matched reference values for lymphocyte subpopulations are generally obtained via cross-sectional studies, whereas patients are followed longitudinally. We performed a detailed longitudinal analysis of the changes in lymphocyte subpopulations in a group of 11 healthy infants followed from birth up to 1 y of age, with special attention for early developmental markers, markers of maturation, and markers of activation. We found that T and B lymphocytes increased at 1 and 6 wk of age, respectively. In contrast, NK cells showed a sharp decline directly after birth, suggesting that they are more important during pregnancy than thereafter. CD45RA+--mainly CD4+--naive T lymphocytes were high at birth, and increased further during the first year of life; they form a large expanding pool of cells, ready for participation in primary immune responses. The absolute counts of CD45RO+ memory T lymphocytes were similar in infants and adults, albeit with a lower level of expression of CD45RO on infant T lymphocytes. Almost all infant T lymphocytes expressed CD38 throughout the first year of life. The abundant expression of CD38 on an infant's T lymphocytes might be related to a greater metabolic need of the large population of naive untriggered cells that are continually involved in primary immune responses during the first year of life. The high B lymphocyte counts in infants mainly concerned CD38+ B lymphocytes throughout the first year of life. Also, the relative frequencies of CD1c+ and CD5+ B lymphocytes were higher throughout the first year of life than in adults. Therefore, CD1c, CD5, and CD38 could be markers of untriggered B lymphocytes. In conclusion, our longitudinal survey of T and B lymphocytes, NK cells, and their subpopulations during the first year of life helps to complete the picture of lymphocyte development in infants. This information contributes to the correct interpretation of data from infants with possible immune disorders.


[Study on relationship between nail-fold microcirculation and histopathology of patients with vocal cord polyp and blood stasis syndrome].
Nail-fold microcirculation of 50 vocal cord polyp patients was observed and compared with hoarseness patients without vocal cord polyp. The results showed that peripheral microcirculation was disturbed in the vocal cord polyp patients. Histopathologic observation of 200 vocal cord polyp patients was made. It was found that principal pathologic change concentrated on the lamina propria mucosae of vocal cord and the most prominent manifestation was disturbance of topical microcirculation of Reinke's layer. In microcirculation aspect, the relationship between polyp of vocal cord and the Blood Stasis Syndrome was confirmed, and pathogenic mechanism of vocal cord polyp was discussed.


Core circadian clock transcription factor BMAL1 regulates mammary epithelial cell growth, differentiation, and milk component synthesis.
The role the mammary epithelial circadian clock plays in gland development and lactation is unknown. We hypothesized that mammary epithelial clocks function to regulate mammogenesis and lactogenesis, and propose the core clock transcription factor BMAL1:CLOCK regulates genes that control mammary epithelial development and milk synthesis. Our objective was to identify transcriptional targets of BMAL1 in undifferentiated (UNDIFF) and lactogen differentiated (DIFF) mammary epithelial cells (HC11) using ChIP-seq. Ensembl gene IDs with the nearest transcriptional start site to ChIP-seq peaks were explored as potential targets, and represented 846 protein coding genes common to UNDIFF and DIFF cells and 2773 unique to DIFF samples. Genes with overlapping peaks between samples (1343) enriched cell-cell adhesion, membrane transporters and lipid metabolism categories. To functionally verify targets, an HC11 line with Bmal1 gene knocked out (BMAL1-KO) using CRISPR-CAS was created. BMAL1-KO cultures had lower cell densities over an eight-day growth curve, which was associated with increased (p<0.05) levels of reactive oxygen species and lower expression of superoxide dismutase 3 (Sod3). RT-qPCR analysis also found lower expression of the putative targets, prolactin receptor (Prlr), Ppara, and beta-casein (Csn2). Findings support our hypothesis and highlight potential importance of clock in mammary development and substrate transport.


Primary urethral reconstruction: the cost minimized approach to the bulbous urethral stricture.
Treatment for urethral stricture disease often requires a choice between readily available direct vision internal urethrotomy (DVIU) and highly efficacious but more technically complex open urethral reconstruction. Using the short segment bulbous urethral stricture as a model, we determined which strategy is less costly. The costs of DVIU and open urethral reconstruction with stricture excision and primary anastomosis for a 2 cm bulbous urethral stricture were compared using a cost minimization decision analysis model. Clinical probability estimates for the DVIU treatment arm were the risk of bleeding, urinary tract infection and the risk of stricture recurrence. Estimates for the primary urethral reconstruction strategy were the risk of wound complications, complications of exaggerated lithotomy and the risk of treatment failure. Direct third party payer costs were determined in 2002 United States dollars. The model predicted that treatment with DVIU was more costly (17,747 dollars per patient) than immediate open urethral reconstruction (16,444 dollars per patient). This yielded an incremental cost savings of $1,304 per patient, favoring urethral reconstruction. Sensitivity analysis revealed that primary treatment with urethroplasty was economically advantageous within the range of clinically relevant events. Treatment with DVIU became more favorable when the long-term risk of stricture recurrence after DVIU was less than 60%. Treatment for short segment bulbous urethral strictures with primary reconstruction is less costly than treatment with DVIU. From a fiscal standpoint urethral reconstruction should be considered over DVIU in the majority of clinical circumstances.


Kidney Transplantation From Hepatitis B Surface Antigen (HBsAg)-Positive Living Donors to HBsAg-Negative Recipients: Clinical Outcomes at a High-Volume Center in China.
Data on kidney transplantation (KTx) from hepatitis B surface antigen (HBsAg)-positive (HBsAg+) donors to HBsAg-negative (HBsAg-) recipients [D(HBsAg+)/R(HBsAg-)] are limited. We aimed to report the outcomes of D(HBsAg+)/R(HBsAg-) KTx in recipients with or without hepatitis B surface antibody (HBsAb). Eighty-three D(HBsAg+)/R(HBsAg-) living KTx cases were retrospectively identified. The 384 cases of KTx from hepatitis B core antibody-positive (HBcAb+) living donors to HBcAb-negative (HBcAb-) recipients [D(HBcAb+)/R(HBcAb-)] were used as the control group. The primary endpoint was posttransplant HBsAg status change from negative to postive (-- →+). Before KTx, 24 donors (28.9%) in the D(HBsAg+)/R(HBsAg-) group were hepatitis B virus (HBV) DNA positive, and 20 recipients were HBsAb-. All 83 D(HBsAg+)/R(HBsAg-) recipients received HBV prophylaxis, while no D(HBcAb+)/R(HBcAb-) recipients received prophylaxis. After a median follow-up of 36 months (range, 6-106) and 36 months (range, 4-107) for the D(HBsAg+)/R(HBsAg-) and D(HBcAb+)/R(HBcAb-) groups, respectively, 2 of 83 (2.41%) D(HBsAg+)/R(HBsAg-) recipients and 1 of 384 (0.26%) D(HBcAb+)/R(HBcAb-) became HBsAg+, accompanied by HBV DNA-positive (P = .083). The 3 recipients with HBsAg-→+ were exclusively HBsAb-/HBcAb- before KTx. Recipient deaths were more frequent in the D(HBsAg+)/R(HBsAg-) group (6.02% vs 1.04%, P = .011), while liver and graft function, rejection, infection, and graft loss were not significantly different. In univariate analyses, pretransplant HBsAb-/HBcAb- combination in the D(HBsAg+)/R(HBsAg-) recipients carried a significantly higher risk of HBsAg-→+, HBV DNA-→+, and death. Living D(HBsAg+)/R(HBsAg-) KTx in HBsAb+ recipients provides excellent graft and patient survivals without HBV transmission. HBV transmission risks should be more balanced with respect to benefits of D(HBsAg+)/R(HBsAg-) KTx in HBsAb-/HBcAb- candidates.


Sonographic signs of early pregnancy.
The early embryo (blastocyst) is implanted about 6 to 7 days after fertilization and becomes completely imbedded within the decidua at 9.5 days. As the exocoelomic cavity (or early gestational sac) enlarges, it becomes visible by ultrasonography. The gestational sac, however, remains within markedly thickened ipsilateral decidua. The uterine cavity remains relatively undistorted (i.e., straight) and can be delineated by ultrasonography. The gestational sac, therefore, can be seen adjacent to a straight uterine cavity within a thickened decidua. This is called an intradecidual sign. Before the appearance of a visible gestational sac, an echogenic area within the thickened decidua may be seen. This is the earliest sign of intrauterine gestation, and it can be seen as early as 25-days menstrual age. The earliest evidence of an embryo is seen about 2 weeks later, when two small bubbles (amniotic and yolk sacs) attach to the wall of the gestational sac. The embryonic disc is located between the two bubbles. This is called the double-bleb sign. The intradecidual and double-bleb signs represent the most important signs of early intrauterine pregnancy.


Altered 8-oxoguanine glycosylase in mild cognitive impairment and late-stage Alzheimer's disease brain.
Eight-hydroxy-2'-deoxyguanosine (8-OHdG) is increased in the brain in late-stage Alzheimer's disease (LAD) and mild cognitive impairment (MCI). To determine if decreased base-excision repair contributes to these elevations, we measured oxoguanine glycosylase 1 (OGG1) protein and incision activities in nuclear and mitochondrial fractions from frontal (FL), temporal (TL), and parietal (PL) lobes from 8 MCI and 7 LAD patients, and 6 age-matched normal control (NC) subjects. OGG1 activity was significantly (P<0.05) decreased in nuclear specimens of FL, TL, and PL in MCI and LAD and in mitochondria from LAD FL and TL and MCI TL. Nuclear OGG1 protein was significantly decreased in LAD FL and MCI and LAD PL. No differences in mitochondrial OGG1 protein levels were found. Overall, our results suggest that decreased OGG1 activity occurs early in the progression of AD, possibly mediated by 4-hydroxynonenal inactivation and may contribute to elevated 8-OHdG in the brain in MCI and LAD.


Changing perspectives on Alzheimer's disease: thinking outside the amyloid box.
The past two decades have seen an explosion in funding and research for Alzheimer's disease (AD), which has resulted in a wealth of data exploring the potential underlying processes, particularly with regard to amyloid-β (Aβ). However, to date, therapies based on this knowledge have not been forthcoming. In seeking an explanation for our current pharmacological failures, it has become clear that amyloid is only one part of a multi-factorial disease process incorporating a wealth of deleterious factors. Additionally, there is strong evidence that the initial production of Aβ is part of the evolutionarily conserved stress response, triggered by a host of upstream factors highly altered in aging. Taken together, these observations place Aβ in a drastically different context, with toxicity occurring secondarily to upstream deleterious factors and rendering current therapeutic strategies oversimplified. This re-conceptualization necessitates a paradigm shift in our scientific and social response to AD, placing a greater emphasis on upstream interventions and public health awareness of the measures that can be taken by most individuals to reduce the risk of AD. With the increasing prevalence of AD and the realization that disease-modifying drugs may not be available in the near future, it is the responsibility of science to better communicate the worth of preventative healthcare measures to the public.


A Systematic Review of Laboratory Evidence for the Abuse Potential of Tramadol in Humans.
Background: Tramadol is an opioid-analgesic that has shown epidemiological evidence of abuse. This review evaluates the evidence for tramadol abuse potential in humans. Methods: A systematic literature search for human abuse liability examinations of tramadol was conducted in September 2018 and yielded 13 total studies. Studies were all within-subject, double-blind, placebo-controlled human laboratory comparisons of tramadol to opioid comparators. Results are organized based upon the route of tramadol administration (oral, parenteral) and the participant population (persons with and without current opioid physical dependence). Outcomes were categorized into self-report ratings of positive and negative effects, observer-ratings of effects, time course of effects, likelihood tramadol was identified as an opioid, and tramadol self-administration. Results: Results indicated the relative abuse potential of tramadol was lower than the opioids to which it was compared. Tramadol produced highest positive effect ratings when administered orally to persons with no opioid physical dependence. Relative to other opioids, it produced substantial negative ratings, generally demonstrated a slower onset of effects, and was less likely to be identified by participants as an opioid, though it did produce a higher rate of self-administration relative to other opioids in the one study reporting that outcome. Results suggest that the abuse potential of tramadol is highest when it is administered orally to non-dependent individuals, and that it likely decreases as the dose increased and when it was administered parentally or to persons with opioid physical dependence. Conclusion: Taken together, individuals may be less likely than with other opioids to escalate tramadol doses, transition from oral to parenteral routes of administration, or continue using tramadol once opioid physical dependence develops. In that way, the human abuse potential of tramadol appears to be different from and lower than other opioid analgesic medications.


Vaping characteristics and expectancies are associated with smoking cessation propensity among dual users of combustible and electronic cigarettes.
Most e-cigarette users who also smoke combustible cigarettes (dual users) begin vaping to quit smoking, yet only a subset succeeds. We hypothesized that reinforcing characteristics of e-cigarettes (vaping reinforcement) would positively predict smoking cessation propensity (SCP) among dual users. Secondary analysis of cross-sectional baseline data from dual users in an ongoing smoking cessation trial. Exploratory and confirmatory factor analysis (EFA and CFA) created latent variables for vaping reinforcement and SCP. A structural equation modeling (SEM) approach was used to test the hypothesis. United States. A national sample of dual users of combustible and electronic cigarettes who smoke and vape at least once per week (n = 2896) were enrolled (63% male; mean age = 29.9 years) into a randomized controlled trial in which they would receive either smoking cessation materials or no smoking cessation materials. Vaping reinforcement was indexed by vaping frequency (days/week vaping, times/day vaping, puffs/e-cigarette use), e-cigarette characteristics [numbers of modifications and tobacco or non-tobacco flavors, nicotine content (mg) and positive e-cigarette expectancies]. SCP was measured by items of confidence, commitment to being smoke-free, cessation motivation (contemplation ladder), change in cigarettes per day since beginning e-cigarette use and negative smoking expectancies. Four factors emerged from the EFA: vaping propensity (vaping frequency, positive expectancies), vaping enthusiasm (e-cigarette modifications, using non-tobacco flavors, puffs per use), nicotine/tobacco flavor (nicotine strength, tobacco flavors) and SCP (negative expectancies about smoking, motivation to quit smoking, reduction in smoking). A CFA upheld the exploratory factor structure [root mean square error of approximation (RMSEA) = 0.046, CFI = 0.91]. An SEM with the three vaping latent variables directly predicting SCP had good model fit (RMSEA = 0.030, CFI = 0.97) with a positive relationship of vaping propensity (0.509, P < 0.001), and small negative relationships of vaping enthusiasm (-0.158, P = 0.014) and nicotine/tobacco flavor (-0.230, P < 0.001). Among e-cigarette users who also smoke combustible cigarettes, frequent vaping combined with positive e-cigarette expectancies appears to predict greater smoking cessation propensity. However, vaping enthusiasm (measured by e-cigarette modifications, using non-tobacco flavors and puffs per use), higher nicotine content and use of tobacco flavored solution may reduce cessation propensity.


Shoulder Internal and External Rotation Peak Torque Production through a Velocity Spectrum in Differing.
The purpose of this study was to explore alternative positions for isokinetic testing of shoulder internal and external rotation. Peak torque of shoulder internal and external rotation was measured with a CybeP dynamometer in 20 normal young right-handed males. Peak torques of subjects' right shoulders were measured in six different shoulder positions and three different velocities (60, 180, and 300 degrees /sec). Each subject made three movements at each of the 18 position/velocity combinations and a mean peak torque was found for each combination. Differences in peak torque for internal and external rotation due to position and velocity were tested separately with a 2-way ANOVA and Duncan Multiple Range Test (p = 0.05). Peak torque values were significantly different among the three velocities for all test positions. Significant differences were found among the six test positions. Although the highest peak torques were significantly greater than the lowest, there were no consistent differences between the Cybex recommended positions and the alternative positions. J Orthop Sports Phys Ther 1987;8(11):518-524.


Dissecting the biology of allogeneic HSCT to enhance the GvT effect whilst minimizing GvHD.
Allogeneic haematopoietic stem cell transplantation (allo-HSCT) was the first successful therapy for patients with haematological malignancies, predominantly owing to graft-versus-tumour (GvT) effects. Dramatic methodological changes, designed to expand eligibility for allo-HSCT to older patients and/or those with comorbidities, have led to the use of reduced-intensity conditioning regimens, in parallel with more aggressive immunosuppression to better control graft-versus-host disease (GvHD). Consequently, disease relapse has become the major cause of death following allo-HSCT. Hence, the prevention and treatment of relapse has come to the forefront and remains an unmet medical need. Despite >60 years of preclinical and clinical studies, the immunological requirements necessary to achieve GvT effects without promoting GvHD have not been fully established. Herein, we review learnings from preclinical modelling and clinical studies relating to the GvT effect, focusing on mechanisms of relapse and on immunomodulatory strategies that are being developed to overcome disease recurrence after both allo-HSCT and autologous HSCT. Emphasis is placed on discussing current knowledge and approaches predicated on the use of cell therapies, cytokines to augment immune responses and dual-purpose antibody therapies or other pharmacological agents that can control GvHD whilst simultaneously targeting cancer cells.


Studies of substrate-induced conformational changes in human cytomegalovirus protease using optical biosensor technology.
The interaction between human cytomegalovirus (HCMV) protease and a peptide substrate was studied using a surface plasmon resonance (SPR)-based biosensor. Immobilization of the enzyme to the sensor chip surface by amine coupling resulted in an active enzyme with a higher catalytic efficiency than the enzyme in solution, primarily due to a lower K(m) value. The interaction between immobilized protease and substrate was characterized by a biphasic SPR signal. Rate constants for the formation of the initial enzyme-substrate complex could be determined from the sensorgrams. Simulated binding curves based on the determined k(cat) and the rate constants indicated that the complex binding signal did not originate from the accumulation of intermediates in the catalytic reaction. By chemical crosslinking of the immobilized HCMV protease, which was shown to limit the enzyme's structural flexibility, it was revealed that the obtained sensorgrams were composed of a signal caused by substrate binding and considerable structural alterations in the immobilized enzyme. Furthermore, HCMV protease was inactivated by chemical crosslinking, indicating that structural flexibility is essential for this enzyme. Parallel experiments with immobilized alpha-chymotrypsin revealed that it does not undergo similar conformational changes on peptide binding and that crosslinking did not inactivate the enzyme. The simultaneous detection of binding and conformational changes using optical biosensor technology is expected to be of importance for further characterization of the enzymatic properties of HCMV protease and for identification of inhibitors of this enzyme. It can also be of use for studies of other flexible proteins.


Tic22 is targeted to the intermembrane space of chloroplasts by a novel pathway.
Tic22 previously was identified as a component of the general import machinery that functions in the import of nuclear-encoded proteins into the chloroplast. Tic22 is peripherally associated with the outer face of the inner chloroplast envelope membrane, making it the first known resident of the intermembrane space of the envelope. We have investigated the import of Tic22 into isolated chloroplasts to define the requirements for targeting of proteins to the intermembrane space. Tic22 is nuclear-endoded and synthesized as a preprotein with a 50-amino acid N-terminal presequence. The analysis of deletion mutants and chimerical proteins indicates that the precursor of Tic22 (preTic22) presequence is necessary and sufficient for targeting to the intermembrane space. Import of preTic22 was stimulated by ATP and required the presence of protease-sensitive components on the chloroplast surface. PreTic22 import was not competed by an excess of an authentic stromal preprotein, indicating that targeting to the intermembrane space does not involve the general import pathway utilized by stromal preproteins. On the basis of these observations, we conclude that preTic22 is targeted to the intermembrane space of chloroplasts by a novel import pathway that is distinct from known pathways that target proteins to other chloroplast subcompartments.


Sub-optimal household water access is associated with greater risk of intimate partner violence against women: evidence from Nepal.
Household water management is often women's responsibility, as related to the gendered nature of household roles. Ethnographic data suggest that household water insecurity could increase women's exposure to emotional and physical forms of intimate partner violence (IPV), as punishments for failures to complete socially expected household tasks that rely on water (like cooking and cleaning) and the generally elevated emotional state of household members dealing with resource scarcity. Here, we test the associations between sub-optimal household water access and women's exposure to IPV, using the nationally-representative data from Nepal Demographic and Health Survey, 2016. Drawing upon the intra-household bargaining model as the theoretical framework, we run instrumental variable probit regression, to test the association between household water access and prevalence of IPV against women. After controlling for other known covariates of IPV such as women's empowerment and education, the findings substantiate that worse household water access consistently elevates women's exposures to all forms of IPV. This suggests that improvements in household water access may have additional ramifications for reducing women's risk of IPV, beyond currently recognized socioeconomic benefits. While both household water access and IPV have known health consequences, linking them provides another pathway through which water could affect women's health.


Phenotypic and Genetic Characteristics of Five Korean Patients with Costello Syndrome.
Costello syndrome (CS) is a rare genetic disorder characterized by distinctive facial appearance, cardiopulmonary complications, severe growth retardation, skin and skeletal defects, developmental delay, and tumor predisposition. CS is caused by heterozygous de novo mutations in the proto-oncogene HRAS, which is a component of the RAS/mitogen-activated protein kinase pathway. Herein, we reviewed the phenotypic and genetic features of 5 Korean patients who were genetically diagnosed with CS. Atrial tachycardia and polyhydramnios, which are important prenatal features for CS, were observed in 4 and 5 patients, respectively. The distinctive coarse facial appearances of the patients and presence of deep palmoplantar creases supported the clinical diagnosis of CS, which was confirmed by HRAS sequence analysis. Extremely poor postnatal growth was observed in all 5 patients. Further, all patients exhibited cardiac abnormalities; left ventricular hypertrophy and hypertrophic cardiomyopathy were observed in 3 patients. All 5 patients suffered from airway problems; 3 of them required intubation right after birth, and 2 of them received tracheostomy. One patient with a p.Gly12Ser mutation was diagnosed with retroperitoneal rhabdomyosarcoma alveolar type at the age of 5 years. Consistent with previous reports, both patients with p.Gly12Cys mutations died within the first year of life due to cardiopulmonary failure. Our study summarizes the characteristics of these 5 Korean patients with CS and, along with previous studies, provides clues for genotype-phenotype correlation in patients with CS.


Altered neuroendocrine status at the onset of CNS lupus-like disease.
Neuropsychiatric (NP) manifestations and brain atrophy are common, etiologically unexplained complications of the systemic autoimmune disease lupus erythematosus (SLE). Similar to patients with NP SLE, behavioral deficits and neurodegeneration occur in aged, lupus-prone MRL/lpr mice. In order to gain a better understanding of the time course and nature of CNS involvement, we compare the neuro-immuno-endocrine profiles of two lupus-prone MRL/lpr stocks, which differ in disease onset and severity. Mice from stock 485 (characterized by early lupus-like manifestations) display blunted responsiveness to palatable solutions and impaired nocturnal activity as early as 7 weeks of age. They also have increased IgG in cerebrospinal fluid (CSF) before high serum autoantibody levels and splenomegaly are detected. Moreover, when compared to age-matched 6825 controls, 485 mice exhibit elevated serum corticosterone, enlarged left adrenal gland, and enhanced haematoxylin/eosin staining in the hypothalamic paraventricular nucleus. Swimming speed and novel object exploration become impaired only when more severe peripheral manifestations are documented in 17 week-old 485 mice. The obtained results suggest that performance deficits during the prodromal phase of NP SLE-like disease are associated with autoantibodies in CSF and asymmetric activation of the hypothalamus-pituitary-adrenal axis. Subsequent deterioration in behavioral performance evolves alongside systemic autoimmunity and inflammation. Although a leaky blood-CSF barrier is a possible explanation, one may hypothesize that, similar to neonatal lupus, maternal antibodies to brain antigens cross blood-placental barrier during embryogenesis and induce early endocrine and behavioral deficits in offspring.


Isolation and characterization of a Na-H antiporter-deficient mutant of LLC-PK1 cells.
LLC-PK1 is an established cell line derived from pig kidney epithelium; it differentiates in vitro to form a polarized epithelial sheet, capable of the vectorial transport of solutes and water. We have used a modification of the "proton-suicide" method of Pouyssegur et al. (Proc. Natl. Acad. Sci. USA. 81:4833-4837, 1984) to isolate a mutant of LLC-PK1 cells that is deficient in Na-H antiporter activity. The mutant grows normally at pH 7.0 and above in the presence and absence of bicarbonate; at pH 6.5, however, it requires bicarbonate for growth. Alkalinization of the cytoplasm by the Na-H antiporter thus appears to be essential for growth at acidic pH in the absence of bicarbonate. The antiporter is also essential for the formation of domes (a consequence of vectorial water transport) in the absence of bicarbonate.


Vector control by insecticide-treated nets in the fight against visceral leishmaniasis in the Indian subcontinent, what is the evidence?
Visceral leishmaniasis (VL) is a deadly vector-borne disease that causes an estimated 500 000 new cases a year. In India, Nepal and Bangladesh, VL is caused by Leishmania donovani, which is transmitted from man to man by the sandfly Phlebotomus argentipes. In 2005, these three countries signed a memorandum of understanding to eliminate VL from the region. Integrated vector management is one of the pillars of this elimination strategy, alongside early case detection and treatment. We reviewed the evidence of effectiveness of different vector control methods, to examine the potential role of insecticide treated bednets (ITNs). Indoor residual spraying has shown poor impact for various reasons and resistance to DDT is emerging in Bihar. Environmental management performed poorly compared to insecticide based methods. ITNs could give individual protection but this still needs to be proven in randomized trials. Given the constraints of indoor residual spraying, it is worthwhile to further explore the use of ITNs, in particular long lasting ITNs, as an additional tool in the VL elimination initiative.


RADIANCE: a randomized controlled study of ranibizumab in patients with choroidal neovascularization secondary to pathologic myopia.
To compare the efficacy and safety of ranibizumab 0.5 mg, guided by visual acuity (VA) stabilization or disease activity criteria, versus verteporfin photodynamic therapy (vPDT) in patients with visual impairment due to myopic choroidal neovascularization (CNV). Phase III, 12-month, randomized, double-masked, multicenter, active-controlled study. Patients (N = 277) with visual impairment due to myopic CNV. Patients were randomized to receive ranibizumab on day 1, month 1, and thereafter as needed guided by VA stabilization criteria (group I, n = 106); ranibizumab on day 1 and thereafter as needed guided by disease activity criteria (group II, n=116); or vPDT on day 1 and disease activity treated with ranibizumab or vPDT at investigators' discretion from month 3 (group III, n = 55). Mean average best-corrected visual acuity (BCVA) change from baseline to month 1 through months 3 (primary) and 6, mean BCVA change and safety over 12 months. Ranibizumab treatment in groups I and II was superior to vPDT based on mean average BCVA change from baseline to month 1 through month 3 (group I: +10.5, group II: +10.6 vs. group III: +2.2 Early Treatment Diabetic Retinopathy Study [ETDRS] letters; both P<0.0001). Ranibizumab treatment guided by disease activity was noninferior to VA stabilization-guided retreatment based on mean average BCVA change from baseline to month 1 through month 6 (group II: +11.7 vs. group I: +11.9 ETDRS letters; P<0.00001). Mean BCVA change from baseline to month 12 was +13.8 (group I), +14.4 (group II), and +9.3 ETDRS letters (group III). At month 12, 63.8% to 65.7% of patients showed resolution of myopic CNV leakage. Patients received a median of 4.0 (group I) and 2.0 (groups II and III) ranibizumab injections over 12 months. No deaths or cases of endophthalmitis and myocardial infarction occurred. Ranibizumab treatment, irrespective of retreatment criteria, provided superior BCVA gains versus vPDT up to month 3. Ranibizumab treatment guided by disease activity criteria was noninferior to VA stabilization criteria up to month 6. Over 12 months, individualized ranibizumab treatment was effective in improving and sustaining BCVA and was generally well tolerated in patients with myopic CNV.


Genome-Wide Identification of Catalase Gene Family and the Function of SmCAT4 in Eggplant Response to Salt Stress.
Salinity is an important abiotic stress, damaging plant tissues by causing a burst of reactive oxygen species (ROS). Catalase (CAT) enzyme coded by Catalase (CAT) genes are potent in reducing harmful ROS and hydrogen peroxide (H2O2) produced. Herein, we performed bioinformatics and functional characterization of four SmCAT genes, retrieved from the eggplant genome database. Evolutionary analysis CAT genes revealed that they are divided into subgroups I and II. The RT-qPCR analysis of SmCAT displayed a differential expression pattern in response to abiotic stresses. All the CAT proteins of eggplant were localized in the peroxisome, except for SmCAT4, which localized in the cytomembrane and nucleus. Silencing of SmCAT4 compromised the tolerance of eggplant to salt stress. Suppressed expression levels of salt stress defense related genes SmTAS14 and SmDHN1, as well as increase of H2O2 content and decrease of CAT enzyme activity was observed in the SmCAT4 silenced eggplants. Our data provided insightful knowledge of CAT gene family in eggplant. Positive regulation of eggplant response to salinity by SmCAT4 provides resource for future breeding programs.


Evolutionary relationships of Drosophila mojavensis geographic host races and their sister species Drosophila arizonae.
The cactophilic Drosophila mojavensis species group living in the deserts and dry tropical forests of the southwestern United States and Mexico provides a valuable system for studies in diversification and speciation. Rigorous studies of the relationships between host races of D. mojavensis and the relationships among the members of the species group (D. mojavensis, Drosophila arizona, and Drosophila navojoa) are lacking. We used mitochondrial CO1 sequence data to address the phylogenetics and population genetics of this species group. In this study we have found that the sister species D. mojavensis and D. arizonae share no mitochondrial haplotypes and thus show no evidence for recent introgression. We estimate the divergence time between D. mojavensis and D. arizonae to be between 1.91 and 2.97 million years ago. D. arizonae shows little structure in our population genetic analyses but there is phylogenetic differentiation between southeastern and northern populations of D. arizonae. Drosophila mojavensis shows significant population and phylogenetic structure across the four geographic regions of its distribution. The mitochondrial data support an origin of D. mojavensis on the mainland with early differentiation into the populations now found in the Mojave Desert and the Mainland Sonoran Desert and later colonization of the Baja Peninsula, in contrast to previous models. Also, the sister clade to D. mojavensis/D. arizonae includes D. navojoa and Drosophila huaylasi. By defining the genetic relationships among these populations, we provide a foundation for more sophisticated hypothesis testing regarding the timing of early speciation events and host switches in this species group.


Drug-tolerant persister cancer cells are vulnerable to GPX4 inhibition.
Acquired drug resistance prevents cancer therapies from achieving stable and complete responses. Emerging evidence implicates a key role for non-mutational drug resistance mechanisms underlying the survival of residual cancer 'persister' cells. The persister cell pool constitutes a reservoir from which drug-resistant tumours may emerge. Targeting persister cells therefore presents a therapeutic opportunity to impede tumour relapse. We previously found that cancer cells in a high mesenchymal therapy-resistant cell state are dependent on the lipid hydroperoxidase GPX4 for survival. Here we show that a similar therapy-resistant cell state underlies the behaviour of persister cells derived from a wide range of cancers and drug treatments. Consequently, we demonstrate that persister cells acquire a dependency on GPX4. Loss of GPX4 function results in selective persister cell ferroptotic death in vitro and prevents tumour relapse in mice. These findings suggest that targeting of GPX4 may represent a therapeutic strategy to prevent acquired drug resistance.


Deep learning for fast super-resolution ultrasound microvessel imaging.
Objective. Ultrasound localization microscopy (ULM) enables microvascular reconstruction by localizing microbubbles (MBs). Although ULM can obtain microvascular images that are beyond the ultimate resolution of the ultrasound (US) diffraction limit, it requires long data processing time, and the imaging accuracy is susceptible to the density of MBs. Deep learning (DL)-based ULM is proposed to alleviate these limitations, which simulated MBs at low-resolution and mapped them to coordinates at high-resolution by centroid localization. However, traditional DL-based ULMs are imprecise and computationally complex. Also, the performance of DL is highly dependent on the training datasets, which are difficult to realistically simulate.Approach. A novel architecture called adaptive matching network (AM-Net) and a dataset generation method named multi-mapping (MMP) was proposed to overcome the above challenges. The imaging performance and processing time of the AM-Net have been assessed by simulation andin vivoexperiments.Main results. Simulation results show that at high density (20 MBs/frame), when compared to other DL-based ULM, AM-Net achieves higher localization accuracy in the lateral/axial direction.In vivoexperiment results show that the AM-Net can reconstruct ∼24.3μm diameter micro-vessels and separate two ∼28.3μm diameter micro-vessels. Furthermore, when processing a 128 × 128 pixels image in simulation experiments and an 896 × 1280 pixels imagein vivoexperiment, the processing time of AM-Net is ∼13 s and ∼33 s, respectively, which are 0.3-0.4 orders of magnitude faster than other DL-based ULM.Significance. We proposes a promising solution for ULM with low computing costs and high imaging performance.


L-Dopa release from mesoporous silica nanoparticles engineered through the concept of drug-structure-directing agents for Parkinson's disease.
Parkinson's disease (PD) is a progressive neurodegenerative disease, the 2nd most common after Alzheimer's disease, the main effect of which is the loss of dopaminergic neurons. Levodopa or l-Dopa is an amino acid used in the treatment of PD that acts as the immediate precursor to dopamine. However, over time the efficacy of the medication gradually decreases requiring modified delivery methods. One of the major challenges for the medication to work is to achieve a gradual continuous supply of l-Dopa to the brain to minimise symptoms. Herein, mesoporous silica nanoparticles (MSNs) were engineered through the concept of drug-structure-directing agents (DSDAs) with inherent therapeutic activity. The DSDA used was l-Dopa drug modified by amidation with fatty acids to build anionic surfactants that were able to form micelles as templates for the assembly of inorganic precursors to form the silica framework. This templating route produced MSNs with tunable sizes ranging from 100 nm to 1 μm and with different shapes: spherical, with either solid structures with radial mesopores and porous shells, or hollow-shells with inside large void cavities; and elongated, characterized by long hollows covered by mesoporous shells. The concept of using DSDAs to synthesize drug nanocarriers can be used to avoid the surfactant removal and subsequent drug loading steps involved in the synthesis of conventional MSNs. We hypothesized that the l-Dopa released from MSN materials is mediated by the size and solubility of the DSDAs, and the surface chemical interactions between the DSDAs and MSN hosts. Different pHs (acidic and neutral) simulating gastrointestinal tract conditions were tested, and the results showed hardly any release for gastric conditions at pH 1.2, avoiding the premature release in the stomach typical of conventional MSNs, while for intestinal conditions of pH 7.4, the release of l-Dopa occurred in a continuous and sustained manner, which is well suited to the drug's application and delivery route, and matches well with achieving a sustained l-Dopa delivery to relief symptoms. This could open up new uses for MSNs synthesized by this approach to treat PD.


POSSUM, p-POSSUM, and Cr-POSSUM: implementation issues in a United States health care system for prediction of outcome for colon cancer resection.
The Physiologic and Operative Severity Score for the enUmeration of Mortality and morbidity (POSSUM), Portsmouth revision (p)-POSSUM, and colorectal (Cr)-POSSUM scoring systems were developed as audit tools for comparing outcomes in surgical and colorectal patients on the basis of operative risk assessment. The aim of this study was to evaluate the applicability of these systems to a cohort of colon cancer patients undergoing surgery in the United States. POSSUM factors from 890 consecutive patients undergoing major surgical procedures for colon cancer in nine United States hospitals over a two-year period from January 2000 through December 2001 were prospectively collected. The observed over the expected hospital mortality was compared by means of the POSSUM, p-POSSUM, and Cr-POSSUM scoring systems. The effect of missing data on the utility of this process for outcome assessment was assessed with three methods for data imputation. The number of resections per institution ranged from 13 to 437. The observed mortality rate ranged from 0.8 percent to 15.4 percent among the institutions, with an overall operative mortality of 2.3 percent. The POSSUM, p-POSSUM, and Cr-POSSUM predicted mortality was 10.7 percent, 11.2 percent, and 4.9 percent, respectively. The POSSUM and p-POSSUM models overpredicted mortality in all institutions ( P < 0.01), whereas the Cr-POSSUM demonstrated an observed over expected hospital mortality ratio of >1 in three institutions. The calculations were unaffected by the various methods of inserting missing data. An apparent overprediction of mortality for colon cancer resection was evident with all three POSSUM variants. This implies that a calibration process is required for use of these variants in the United States health care system. Missing data may be treated as normal values without influencing outcome. The Cr-POSSUM appeared to be the most promising audit tool for colorectal cancer surgery; however, it will require further refinement to provide process control graphs for identification of potential outliers and improvement in the quality of care in the United States.


Robot-guided pediatric stereoelectroencephalography: single-institution experience.
OBJECTIVEStereoelectroencephalography (SEEG) has increased in popularity for localization of epileptogenic zones in drug-resistant epilepsy because safety, accuracy, and efficacy have been well established in both adult and pediatric populations. Development of robot-guidance technology has greatly enhanced the efficiency of this procedure, without sacrificing safety or precision. To date there have been very limited reports of the use of this new technology in children. The authors present their initial experience using the ROSA platform for robot-guided SEEG in a pediatric population.METHODSBetween February 2016 and October 2017, 20 consecutive patients underwent robot-guided SEEG with the ROSA robotic guidance platform as part of ongoing seizure localization and workup for medically refractory epilepsy of several different etiologies. Medical and surgical history, imaging and trajectory plans, as well as operative records were analyzed retrospectively for surgical accuracy, efficiency, safety, and epilepsy outcomes.RESULTSA total of 222 leads were placed in 20 patients, with an average of 11.1 leads per patient. The mean total case time (± SD) was 297.95 (± 52.96) minutes and the mean operating time per lead was 10.98 minutes/lead, with improvements in total (33.36 minutes/lead vs 21.76 minutes/lead) and operative (13.84 minutes/lead vs 7.06 minutes/lead) case times/lead over the course of the study. The mean radial error was 1.75 (± 0.94 mm). Clinically useful data were obtained from SEEG in 95% of cases, and epilepsy surgery was indicated and performed in 95% of patients. In patients who underwent definitive epilepsy surgery with at least a 3-month follow-up, 50% achieved an Engel class I result (seizure freedom). There were no postoperative complications associated with SEEG placement and monitoring.CONCLUSIONSIn this study, the authors demonstrate that rapid adoption of robot-guided SEEG is possible even at a SEEG-naïve institution, with minimal learning curve. Use of robot guidance for SEEG can lead to significantly decreased operating times while maintaining safety, the overall goals of identification of epileptogenic zones, and improved epilepsy outcomes.


Analysis of sequence diversity through internal transcribed spacers and simple sequence repeats to identify Dendrobium species.
The Orchidaceae is one of the largest and most diverse families of flowering plants. The Dendrobium genus has high economic potential as ornamental plants and for medicinal purposes. In addition, the species of this genus are able to produce large crops. However, many Dendrobium varieties are very similar in outward appearance, making it difficult to distinguish one species from another. This study demonstrated that the 12 Dendrobium species used in this study may be divided into 2 groups by internal transcribed spacer (ITS) sequence analysis. Red and yellow flowers may also be used to separate these species into 2 main groups. In particular, the deciduous characteristic is associated with the ITS genetic diversity of the A group. Of 53 designed simple sequence repeat (SSR) primer pairs, 7 pairs were polymorphic for polymerase chain reaction products that were amplified from a specific band. The results of this study demonstrate that these 7 SSR primer pairs may potentially be used to identify Dendrobium species and their progeny in future studies.


Interleukin-6 directly influences proliferation and invasion potential of head and neck cancer cells.
Interleukin-6 (IL-6) is a multifunctional regulator of immune response and hematopoiesis. Recently, it has been reported that expression of IL-6 is correlated with prognosis in various cancer patients. In this study, we investigated whether the proliferation and invasion potential of head and neck squamous cell carcinomas (HNSCCs) were influenced by IL-6. All HNSCC cell lines, HEp-2, HSC-2, HSC-4, and SAS, were tested by reverse transcription-polymerase chain reaction (RT-PCR) and expressed the IL-6 receptor (IL-6R), and glycoprotein 130, which is responsible for signal transduction. HEp-2, HSC-2, and HSC-4 also produced IL-6. IL-6 inhibited the proliferation of HSC-2 and SAS, but the invasion potential of all the cell lines increased. Moreover, IL-6 down-regulated soluble IL-6R expression. Anti-IL-6R antibody abrogated the inhibited proliferation and increased invasion induced by IL-6. IL-6 stimulation also induced the extracellular regulated protein kinase 1/2 activation and increased vascular endothelial growth factor release. These results suggest that IL-6 can directly influence cell proliferation and the invasion potential as the first step of tumor metastasis.


α-Synuclein-specific T cell reactivity is associated with preclinical and early Parkinson's disease.
A diagnosis of motor Parkinson's disease (PD) is preceded by a prolonged premotor phase with accumulating neuronal damage. Here we examined the temporal relation between α-synuclein (α-syn) T cell reactivity and PD. A longitudinal case study revealed that elevated α-syn-specific T cell responses were detected prior to the diagnosis of motor PD, and declined after. The relationship between T cell reactivity and early PD in two independent cohorts showed that α-syn-specific T cell responses were highest shortly after diagnosis of motor PD and then decreased. Additional analysis revealed significant association of α-syn-specific T cell responses with age and lower levodopa equivalent dose. These results confirm the presence of α-syn-reactive T cells in PD and show that they are most abundant immediately after diagnosis of motor PD. These cells may be present years before the diagnosis of motor PD, suggesting avenues of investigation into PD pathogenesis and potential early diagnosis.


Brachybacterium muris sp. nov., isolated from the liver of a laboratory mouse strain.
A coccoid- to ovoid-shaped, Gram-positive bacterial strain, designated C3H-21(T), was isolated from the liver of the laboratory mouse strain C3H/He and characterized by a polyphasic approach. The peptidoglycan type was variation A4gamma with meso-diaminopimelic acid as the diagnostic cell-wall diamino acid and an interpeptide bridge of D-asp-D-Glu. The isolate contained menaquinone MK-7 (88 %) as the major component of the quinone system and minor amounts of menaquinone MK-8 (9 %) and menaquinone MK-6 (3 %). The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, unidentified glycolipids and unidentified phospholipids. The fatty acid profile contained predominantly anteiso-C(15 : 0) and significant amounts of iso-C(16 : 0), iso-C(14 : 0,) anteiso-C(17 : 0) and C(19 : 0). The polyamine pattern consisted of spermine and spermidine as the major compounds. Genomic fingerprints clearly distinguished strain C3H-21(T) from other Brachybacterium species. The isolate shared the highest 16S rDNA sequence similarities with members of the genus Brachybacterium, in particular Brachybacterium sacelli LMG 20345(T), Brachybacterium nesterenkovii DSM 9573(T), Brachybacterium rhamnosum LMG 19848(T), Brachybacterium alimentarium CNRZ 925(T) and Brachybacterium fresconis LMG 20336(T) (97.8-97.2 %). The results of biochemical/physiological characterization, chemotaxonomic characteristics and REP-PCR-generated fingerprints demonstrated that the isolate represents a novel species of the genus Brachybacterium, for which the name Brachybacterium muris (type strain C3H-21(T)=DSM 15460(T)=CCM 7047(T)) [corrected] is proposed.