Functional analysis of the chimpanzee and human apo(a) promoter sequences: identification of sequence variations responsible for elevated transcriptional activity in chimpanzee.
Lp(a) concentrations vary considerably among individuals and are primarily determined by the apo(a) gene locus. We have previously shown that mean plasma Lp(a) levels in the chimpanzee are significantly higher than those observed in humans (Doucet, C., Huby, T., Chapman, J., and Thillet, J. (1994) J. Lipid Res 35, 263-270). To evaluate the possibility that this difference may result from a high level of expression of chimpanzee apo(a), we cloned and sequenced 1.4 kilobase (kb) of the 5'-flanking region of the gene and compared promoter activity to that of its human counterpart. Sequence analysis revealed 98% homology between chimpanzee and human apo(a) 5' sequences; among the differences observed, two involved polymorphic sites associated with Lp(a) levels in humans. The TTTTA repeat located 1.3 kb 5' of the apo(a) gene, present in a variable number of copies (n = 5-12) in humans, is uniquely present as four copies in the chimpanzee sequence. The second position concerns the +93 C>T polymorphism that creates an additional ATG start codon in the human apo(a) gene, thereby impairing translation efficiency. In chimpanzee, this position did not appear polymorphic, and a base difference at position +94 precluded the presence of an additional ATG. In transient transfection assays, the chimpanzee apo(a) promoter exhibited a 5-fold elevation in transcriptional activity as compared with its human counterpart. This marked difference in activity was maintained with either 1.4 kb of 5' sequence or the minimal promoter region -98 to +141 of the human and chimpanzee apo(a) genes. Using point mutational analyses, nucleotides present at positions -3, -2, and +8 (relative to the start site of transcription) were found to be essential for the high transcription efficiency of the chimpanzee apo(a) promoter. High transcriptional activity of the chimpanzee apo(a) gene may therefore represent a key factor in the elevated plasma Lp(a) levels characteristic of this non-human primate.


Plasticity and redundancy among AMA-RON pairs ensure host cell entry of Toxoplasma parasites.
Malaria and toxoplasmosis are infectious diseases caused by the apicomplexan parasites Plasmodium and Toxoplasma gondii, respectively. These parasites have developed an invasion mechanism involving the formation of a moving junction (MJ) that anchors the parasite to the host cell and forms a ring through which the parasite penetrates. The composition and the assembly of the MJ, and in particular the presence of protein AMA1 and its interaction with protein RON2 at the MJ, have been the subject of intense controversy. Here, using reverse genetics, we show that AMA1, a vaccine candidate, interacts with RON2 to maintain the MJ structural integrity in T. gondii and is subsequently required for parasite internalization. Moreover, we show that disruption of the AMA1 gene results in upregulation of AMA1 and RON2 homologues that cooperate to support residual invasion. Our study highlights a considerable complexity and molecular plasticity in the architecture of the MJ.


The calcitonin receptor on T 47D breast cancer cells. Evidence for glycosylation.
The glycosyl nature of the receptor for the peptide hormone calcitonin has been investigated in a human breast cancer cell line, T 47D. Studies have been carried out to assess the ability of various lectins and of the antibiotic tunicamycin to inhibit specific binding of calcitonin to the cells, to reduce cross-linking of photoactive calcitonin to a macromolecular receptor component and to influence calcitonin stimulation of cyclic AMP. Pre-incubation of cells with low concentrations of tunicamycin for 72 h resulted in a reduction of total specific binding by approx. 80% and a 40% reduction in calcitonin-stimulated adenylate cyclase; formation of the cross-linked receptor component was also inhibited. Wheat-germ lectin showed the most marked inhibition of total specific binding and cyclic AMP production. However, cross-linking of photoactive calcitonin to receptor component was totally inhibited by this lectin. Soya-bean lectin brought about very little reduction in total specific binding but had more profound effects on calcitonin-stimulated cyclic AMP production and cross-linking of photoactive calcitonin. Concanavalin A and lentil lectin showed some inhibition of all parameters. The data indicate that the calcitonin receptor in T 47D cells is associated with glycosyl moieties, the major contributors of which are N-acetyl-D-glucosamine residues, but N-acetyl-D-galactosamine and mannose residues are also associated.


A triadic neurocognitive approach to addiction for clinical interventions.
According to the triadic neurocognitive model of addiction to drugs (e.g., cocaine) and non-drugs (e.g., gambling), weakened "willpower" associated with these behaviors is the product of an abnormal functioning in one or more of three key neural and cognitive systems: (1) an amygdala-striatum dependent system mediating automatic, habitual, and salient behaviors; (2) a prefrontal cortex dependent system important for self-regulation and forecasting the future consequences of a behavior; and (3) an insula dependent system for the reception of interoceptive signals and their translation into feeling states (such as urge and craving), which in turn plays a strong influential role in decision-making and impulse control processes related to uncertainty, risk, and reward. The described three-systems account for poor decision-making (i.e., prioritizing short-term consequences of a decisional option) and stimulus-driven actions, thus leading to a more elevated risk for relapse. Finally, this article elaborates on the need for "personalized" clinical model-based interventions targeting interactions between implicit processes, interoceptive signaling, and supervisory function aimed at helping individuals become less governed by immediate situations and automatic pre-potent responses, and more influenced by systems involved in the pursuit of future valued goals.


Doc practice management set to explode.
An industry that barely existed give years ago is emerging as a prime mechanism to organize doctors. Independent physician management companies are uniquely situated to do the job as HMOs rethink the strategy of employing physicians and as hospitals find doctors reluctant to come under their control.


[Use of immunoglobulin E and pregnancy-specific beta-1-glycoprotein in differential diagnosis of bone malignancies].
Such factors as high levels of immunoglobulin E and pregnancy-specific 1-glycoprotein (PSG) appear as diagnostic criteria for both bone malignancies and acute hematogenous osteomyelitis. It was found that significant increase in blood-serum PSG in conjunction with lowered IgE is a marker of bone malignancy while decreased PSG and high IgE is that of acute hematogenous osteomyelitis.


Cytoplasmic zone analysis. RNA flow studied by micromanipulation.
A technique is described, cytoplasmic zone analysis, by which it is possible to study the flow of different RNA-containing components in the cytoplasm after their release from the nucleus. After a pulse of RNA precursors, the salivary glands of the insect Chironomus tentans are fixed and microdissected for the isolation of three zones of cytoplasm situated at increasing distances from the nucleus. The RNA from each zone is isolated and analyzed by gel electrophoresis. The three ribosomal RNA components, 18 S, 28 S and 5 S RNA, appear in steep, specific radioactivity gradients (exit gradients) during the time interval 2-3 h after a precursor injection, the 18 S RNA gradient lying 30-50 mum peripheral to that of the 28 S or 5 S RNA gradient. Administration of puromycin led to the complete disappearance of the 28 S RNA and most of the 28 S RNA gradient within 45 min, suggesting that the gradients are caused by an engagement of the ribosomal subunits into polysomes close to the nucleus immediately or soon after their exit from the nucleus. The gradients may offer a unique model for the study of polysome formation and maintenance in vivo.


Breastfeeding and maternal health outcomes: a systematic review and meta-analysis.
To evaluate the effect of breastfeeding on long-term (breast carcinoma, ovarian carcinoma, osteoporosis and type 2 diabetes mellitus) and short-term (lactational amenorrhoea, postpartum depression, postpartum weight change) maternal health outcomes. A systematic literature search was conducted in PubMed, Cochrane Library and CABI databases. Outcome estimates of odds ratios or relative risks or standardised mean differences were pooled. In cases of heterogeneity, subgroup analysis and meta-regression were explored. Breastfeeding >12 months was associated with reduced risk of breast and ovarian carcinoma by 26% and 37%, respectively. No conclusive evidence of an association between breastfeeding and bone mineral density was found. Breastfeeding was associated with 32% lower risk of type 2 diabetes. Exclusive breastfeeding and predominant breastfeeding were associated with longer duration of amenorrhoea. Shorter duration of breastfeeding was associated with higher risk of postpartum depression. Evidence suggesting an association of breastfeeding with postpartum weight change was lacking. This review supports the hypothesis that breastfeeding is protective against breast and ovarian carcinoma, and exclusive breastfeeding and predominant breastfeeding increase the duration of lactational amenorrhoea. There is evidence that breastfeeding reduces the risk of type 2 diabetes. However, an association between breastfeeding and bone mineral density or maternal depression or postpartum weight change was not evident.


Trans-Differentiation of Human Dental Pulp Stem Cells Into Cholinergic-Like Neurons Via Nerve Growth Factor.
Cell therapy has been widely considered as a therapeutic approach for neurodegenerative diseases and nervous system damage. Cholinergic neurons as one of the most important neurons that play a significant role in controlling emotions, mobility, and autonomic systems. In this study, Human Dental Pulp Stem Cells (hDPSCs) were differentiated into the cholinergic neurons by β-mercaptoethanol in the preinduction phase and also by the nerve growth factor (NGF) in the induction phase. The hDPSCs were evaluated for CD73, CD31, CD34, and Oct-4. Concentration-time relationships for NGF were assessed by evaluating the viability rate of cells and the immune response to nestin, neurofilament 160, microtubule-associated protein-2, and choline acetyltransferase. The hDPSCs had a negative response to CD34 and CD31. The optimal dose for the NGF was 50 ng/mL seven days after the induction when the highest percentage of expressing markers for the Cholinergic neurons (ChAT) was detected. The results of this study provided a method for producing cholinergic neurons by hDPSCs, which can be used in cytotherapy for degenerative diseases of the nervous system and also spinal cord injury.


Drf1-dependent kinase interacts with Claspin through a conserved protein motif.
The Dbf4/Drf1-dependent kinase (DDK) is required for the initiation of DNA replication in eukaryotes. Another protein, Claspin, mediates the activation of a cellular checkpoint response to stalled replication forks and is also a regulator of replication. In this study, we found that DDK phosphorylates Claspin in vitro and forms a nuclear complex containing Cdc7, Drf1, and Claspin in Xenopus egg extracts. In addition, purified Claspin and DDK are capable of a direct in vitro interaction. We identified a conserved binding site on Claspin required for its interaction with DDK. This site corresponds to the first of two sequence repeats in the Chk1-binding domain of Claspin. Furthermore, we have established that two amino acids in this motif, Asp(861) and Gln(866), are essential for the interaction between Claspin and DDK. We found that mutant forms of Claspin incapable of interacting with DDK are still able to associate with and activate Chk1 in response to DNA replication blockages. However, Claspin-depleted egg extracts that have been reconstituted with these mutants of Claspin undergo DNA replication more slowly. These findings suggest that the interaction of DDK with Claspin mediates a checkpoint-independent function of Claspin related to DNA replication.


Improvement of microbial assays of vitamins.
A method for the improvement of microbial assays of vitamins, which involves the addition of a surfactant to the incubated test, was developed. This surfactant tends to eliminate bacterial clumping, giving a uniform suspension of single cells, thereby making the turbidity readings less erratic and the actual assay standard curves more closely related to the desired theoretical curve.


Relationship Between Cytogenetic Complexity and Peritumoral Edema in High-Grade Astrocytoma.
The purpose of the study is to reveal the association of cytogenetic compltyexi and peritumoral edema volume (PTEV) and its prognostic significance in high-grade astrocytoma patients by culturing patient tumor cells. Twenty-seven high-grade astrocytoma patients were divided into three groups according to karyotype complexity: normal, non-complex karyotype (NCK), and complex karyotype (CK). Endothelial growth factor receptor (EGFR) amplification was detected by FISH, and its association with chromosome 7 abnormalities was analyzed. Mean PTEV of each group was compared by ANOVA to evaluate the relationship between PTEV and cytogenetic complexity. The PTEV of patients in normal (n=6), NCK (n=8), and CK (n=13) groups were 24.52±17.73, 34.26±35.04, and 86.31±48.7 cm³, respectively (P=0.005). Ten out of 11 patients with EGFR amplification showed abnormalities in chromosome 7. The mean PTEV of EGFR-amplified and non-amplified groups were 80.4±53.7 and 41.3±37.9 cm³, respectively (P=0.035). The average survival of patients with PTEV less than 90 cm³ was 30.52±26.11 months, while in patients with PTEVs over or equal to 90 cm³, it was 10.83±5.53 months (P=0.007). The results show an association of complex karyotype with the PTEV of high-grade astrocytoma. EGFR amplification plays a significant role in the formation of peritumoral edema, causing PTEV to increase, which is related with survival. This implies that cytogenetic karyotype can be applied as a prognostic factor.


Carpal tunnel syndrome and work organisation in repetitive work: a cross sectional study in France. Study Group on Repetitive Work.
To study the determinants of signs of carpal tunnel syndrome (CTS) in repetitive industrial work, with special attention to occupational constraints at group level and management practices of the companies. A cross sectional study was conducted in three sectors: assembly line; clothing and shoe industry; food industry. A total of 1210 workers in repetitive work, from 53 different companies, was compared with a control group of 337 workers. Constraints at the workplace were partly self declared, and partly assessed by the occupational physicians in charge of the employees of the company. The definition of CTS was based on a standardised clinical examination. CTS was associated with repetitive work, especially packaging. It was more frequent among subjects who declared psychological and psychosomatic problems and those with a body mass index > or = 27. Dissatisfaction with work, lack of job control, short cycle time, and having to press repeatedly with the hand were associated with the syndrome. An odds ratio (OR) of 2.24 was found for "just in time" production. The results emphasise the complexity of the determinants of CTS, the role of psychosocial factors at work and the potentially negative effects of some practices of the companies aimed at enhancing their competitiveness.


The genetic architecture of UV floral patterning in sunflower.
The patterning of floral ultraviolet (UV) pigmentation varies both intra- and interspecifically in sunflowers and many other plant species, impacts pollinator attraction, and can be critical to reproductive success and crop yields. However, the genetic basis for variation in UV patterning is largely unknown. This study examines the genetic architecture for proportional and absolute size of the UV bullseye in Helianthus argophyllus , a close relative of the domesticated sunflower. A camera modified to capture UV light (320-380 nm) was used to phenotype floral UV patterning in an F 2 mapping population, then quantitative trait loci (QTL) were identified using genotyping-by-sequencing and linkage mapping. The ability of these QTL to predict the UV patterning of natural population individuals was also assessed. Proportional UV pigmentation is additively controlled by six moderate effect QTL that are predictive of this phenotype in natural populations. In contrast, UV bullseye size is controlled by a single large effect QTL that also controls flowerhead size and co-localizes with a major flowering time QTL in Helianthus . The co-localization of the UV bullseye size QTL, flowerhead size QTL and a previously known flowering time QTL may indicate a single highly pleiotropic locus or several closely linked loci, which could inhibit UV bullseye size from responding to selection without change in correlated characters. The genetic architecture of proportional UV pigmentation is relatively simple and different from that of UV bullseye size, and so should be able to respond to natural or artificial selection independently.


Regulation of glutamine production by skeletal muscle cells in culture.
Cells of the L6 rat myogenic line consume glutamine in contrast to skeletal muscle in vivo, which produces glutamine. The basis of glutamine consumption in the cultured cells and the conditions under which net glutamine production occurs were investigated. After treating cultures with cytosine-1-beta-D-arabinofuranoside to eliminate replicating myoblasts remaining after differentiation, the L6 cells continued to consume glutamine from medium containing normal plasma levels (650 microM). Consumption declined when cells were incubated with 325 microM glutamine, and, at 0 microM, net production occurred. In addition to the tendency to consume glutamine, the cells rapidly consumed glutamic acid. When medium glutamic acid was increased from 70 to 150, 500, and 1,000 microM, glutamine production increased. Alterations in levels of NH3, the other substrate for glutamine synthesis, did not have this effect. The L6 cells were found to have glutamine synthetase activity similar to normal rat skeletal muscle but markedly increased glutaminase activity. In spite of the imbalance in synthetic and degradative activities, however, inhibitor studies with L-methionine sulfoximine and 6-diazo-5-oxo-L-norleucine showed that flux through both pathways influenced net glutamine production.


[Not Available].
M. Pocard, J.-C. Sabourin In theory, the concept of sentinel lymph node (SLN) biopsy can be applied to cancer surgery for all solid cancers. Yet sentinel lymph node biopsy has not become a standard part of gastrointestinal cancer surgery. It has been of value in the assessment of small early-stage gastric cancers, but has only achieved widespread practice in Japan. Studies of SLN biopsy in colon cancer have not shown it to be a reliable predictor of N+ status and therefore don't permit the omission of lymph node dissection in selected cases. On the other hand, as a form of intra-operative lymph node mapping, dye injection of the SLN may demonstrate aberrant lymphatic drainage and could occasionally permit the sparing of a middle colic artery whose sacrifice would otherwise be dictated by standard drainage patterns. SLN biopsy may have prognostic usefulness by demonstrating micrometastases; careful serial sectioning focussed on the SLN may detect nests of metastatic cells on HE staining, thereby converting a tumor from Stage I (TxN0M0) to Stage II (TxN1M0). This finding has been noted in 10-15% of cases. However, the prognostic value of micrometastases detected only by immunohistochemical staining or PCR has not been demonstrated. For cancers of the anal canal, SLN biopsy of inguinal nodes has been tested as a means of establishing the indications for inguinal lymph node dissection.


Identification and characterization of avihepadnaviruses isolated from exotic anseriformes maintained in captivity.
Five new hepadnaviruses were cloned from exotic ducks and geese, including the Chiloe wigeon, mandarin duck, puna teal, Orinoco sheldgoose, and ashy-headed sheldgoose. Sequence comparisons revealed that all but the mandarin duck viruses were closely related to existing isolates of duck hepatitis B virus (DHBV), while mandarin duck virus clones were closely related to Ross goose hepatitis B virus. Nonetheless, the S protein, core protein, and functional domains of the Pol protein were highly conserved in all of the new isolates. The Chiloe wigeon and puna teal hepatitis B viruses, the two new isolates most closely related to DHBV, also lacked an AUG start codon at the beginning of their X open reading frame (ORF). But as previously reported for the heron, Ross goose, and stork hepatitis B viruses, an AUG codon was found near the beginning of the X ORF of the mandarin duck, Orinoco, and ashy-headed sheldgoose viruses. In all of the new isolates, the X ORF ended with a stop codon at the same position. All of the cloned viruses replicated when transfected into the LMH line of chicken hepatoma cells. Significant differences between the new isolates and between these and previously reported isolates were detected in the pre-S domain of the viral envelope protein, which is believed to determine viral host range. Despite this, all of the new isolates were infectious for primary cultures of Pekin duck hepatocytes, and infectivity in young Pekin ducks was demonstrated for all but the ashy-headed sheldgoose isolate.


Ranibizumab for exudative age-related macular degeneration: A five year study of adherence to follow-up in a real-life setting.
To analyze adherence to follow-up over 5 years in patients treated with intravitreal ranibizumab for exudative age-related macular degeneration (AMD) in a tertiary health care center. To investigate factors associated with failure to continue follow-up. Retrospective chart review of all consecutive patients with exudative AMD, who received their first intravitreal ranibizumab injection at the Créteil Intercommunal University Hospital Eye Clinic between October 1, 2006 and March 31, 2007. Patient clinical characteristics at baseline and at the last follow-up visit were recorded. Distance from home to hospital was measured for each patient. A multiple-choice telephone survey was conducted for patients lost to follow-up to determine the main reasons for failure to continue follow-up. Two hundred and one patients were included. The rate of loss to follow-up over the 5-year period was 57% (115/201). Fifty-eight patients lost to follow-up completed the questionnaire. The main reasons reported by patients for follow-up discontinuation were long distance from home to hospital (51.7%, 30/58), subjective dissatisfaction with the benefits of intravitreal injections (34.5%, 20/58), and the excessive burden of periodic follow-up visits (24.1%, 14/58). Three factors were significantly associated with follow-up discontinuation: high age at baseline (82.2 vs. 76.5 years, P<0.001), poor best-corrected visual acuity (BCVA) at baseline (42.5 vs. 51.0 letters, P=0.020), and long distance from home to hospital (132 vs. 17.1km, P<0.001). In this study, adherence to follow-up over 5 years was poor. Age and BCVA at baseline and distance from home to hospital were independently associated with long-term adherence.


Prevention and promotion effects of Self Help Plus: secondary analysis of cluster randomised controlled trial data among South Sudanese refugee women in Uganda.
Evidence-based and scalable prevention and promotion focused mental health and psychosocial support interventions are needed for conflict-affected populations in humanitarian settings. This study retrospectively assessed whether participation in Self Help Plus (SH+) versus enhanced usual care (EUC) resulted in reduced incidence of probable mental disorder and increased positive mental health and well-being post-intervention among South Sudanese refugee women in Uganda. This study used secondary data from treatment-oriented pilot (n=50) and fully-powered cluster randomised controlled trials (cRCT)s (n=694) of SH+ versus EUC. Data from baseline and post-intervention assessments were combined. A composite latent indicator for mental health problems was generated using mental health and well-being measures included in both cRCTs. In order to assess incidence, a binary variable approximating probable mental disorder was created to exclude those with probable mental disorder from the analysis sample and as the primary prevention outcome. The promotive effects of SH+ relative to EUC were examined in the same sample by assessing subjective well-being and psychological flexibility scale scores. A single factor for mental health problems was identified with all factor loadings >0.30 and acceptable internal consistency (α=0.70). We excluded 161 women who met criteria for probable mental disorder at baseline. Among those with at least moderate psychological distress but without probable mental disorder at baseline and with follow-up data (n=538), the incidence of probable mental disorder at post-intervention was lower among those who participated in SH+ relative to EUC (Risk ratio =0.16, 95% CI: 0.05 to 0.53). Participation in SH+ versus EUC was also associated with increased subjective well-being (β=2.62, 95% CI: 1.63 to 3.60) and psychological flexibility (β=4.55, 95% CI: 2.92 to 6.18) at post-intervention assessment. These results support the use and further testing of SH+ as a selective and indicated prevention and promotion focused psychosocial intervention in humanitarian settings. ISRCTN50148022.


Detection of SYT-SSX1/2 fusion transcripts by reverse transcriptase-polymerase chain reaction (RT-PCR) is a valuable diagnostic tool in synovial sarcoma.
Cytogenetically, most synovial sarcomas are characterised by a specific chromosomal translocation [(X;18) (p11.2;q11.2)], which results in the generation of fusion transcripts comprising SYT (18q11) and either SSX1 or SSX2 (Xp11) sequences. By using a sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) protocol, specific SYT-SSX1/2 fusion transcripts were detected in 10 histopathologically confirmed synovial sarcomas. Control tumours with morphological spindle cell patterns mimicking monophasic synovial sarcoma tested negative (18/19) in the RT-PCR protocol, with the exception of one spindle cell sarcoma originally classified as a fibrosarcoma. Furthermore, the established RT-PCR protocol was used to evaluate the feasibility of SYT-SSX1/2 fusion transcript detection for minimal residual disease analysis. Analyses of surgical margins revealed a fusion transcript in two of four operations for synovial sarcoma analysed, one of which was diagnosed with tumour free margins by conventional histopathology. These data suggest that the RT-PCR amplification of SYT-SSX1/2 fusion transcripts is a valuable tool in the differentiation of synovial sarcomas, especially in cases of equivocal morphology. Additionally, the RT-PCR approach may be used for the detection of residual tumour cells in synovial sarcoma patients.


Percutaneous fixation of tibial plateau fractures under arthroscopy: a medium term perspective.
Arthroscopically assisted percutaneous internal fixation has found its place in the treatment of Schatzker I-III tibial plateau fractures, with good short-term results reported. The objective of this study was to observe the progression of osteoarthritis at the medium term through clinical and radiological assessment. Twenty-seven patients were treated with arthroscopy-assisted percutaneous fixation for stage I-III Schatzker tibial plateau fractures. Twenty-one patients were reviewed with a mean follow-up of 59.5 months (range, 24-138 months); satisfaction was good except for return to sports activity. The mean IKS score was 85.2 for the knee score and 91 for function. The mean Lysholm score was 86 points, with a mean Tegner activity score of 4. A mean score of 25.5 and 8 points was found for the clinical and radiological Rasmussen scores, respectively; 47.6% of the patients presented early osteoarthritis on radiological evaluation. The medium-term functional results were comparable to the short-term results. The patients were satisfied except for return to sports activity. Age at surgery appears as a prognostic factor for osteoarthritis. Arthroscopic internal fixation remains the technical reference for Schatzker I-III tibial plateau fractures despite the appearance of osteoarthritis, which remains less extensive than in open surgery. Level IV. Retrospective study.


A possible role of NF-kappaB and HSP72 in skeletal muscle hypertrophy induced by heat stress in rats.
Effects of heat stress on phosphorylated nuclear factor-kappaB (phospho-NF-kappaB) and tumor necrosis factor alpha (TNFalpha) contents in skeletal muscles were studied. Male Wistar rats (7-week-old) were randomly assigned to control and heat-stressed groups. Rats in heat-stressed group were exposed to heat stress (42 degrees C for 60 min) in an incubator without anesthesia. Soleus muscles were dissected and weighted 1, 3, and 7 days after the heat exposure. Significant increases in the wet weight and protein content of soleus were observed 7 days following the exposure (p < 0.05). Heat stress also induced the up-regulation of heat shock protein 72 (HSP72), IkappaBalpha (inhibitor of NF-kappaB) and the increase in the relative population of Pax7-positive satellite cells to total muscle nuclei before the increase in muscle mass. The content levels of phospho-NF-kappaB and TNFalpha were significantly decreased 1 and 3 days after heat stress, respectively (p < 0.05). A negative correlation between HSP72 and phospho-NF-kappaB contents was observed 1 day after the heat exposure. These observations suggest that the decrease in NF-kappaB signaling may play a part of a role in heat stress-associated muscle hypertrophy.


IFN-gamma: a cytokine essential for rejection of CTL-resistant, virus-infected cells.
We recently demonstrated differential susceptibility of cells expressing viral antigen to killing by antigen-specific cytotoxic T lymphocytes (CTLs). In addition, interferon-gamma (IFN-gamma) has been implicated in the clearance of some viruses from tissues. We explored the role of IFN-gamma in the cytotoxicity of Sendai virus-specific CTLs against virus-infected RL(male symbol)1 (T cell leukemia) or Meth A (fibrosarcoma) cells, as well as the growth of subcutaneously (s.c.) transplanted, virus-infected cells in IFN-gamma(+/+) or IFN-gamma(/) mice of the syngeneic strain (BALB/c). Sendai virus-specific CTLs were cytotoxic against virus-infected RL(male symbol)1 cells, and s.c. transplanted, virus-infected RL(male symbol)1 cells were acutely rejected from IFN-gamma(+/+) or IFN-gamma(/) mice. In contrast, the CTLs were inactive toward virus-infected Meth A cells, but s.c. transplanted, virus-infected Meth A cells were acutely rejected from IFN-gamma(+/+) but not IFN-gamma(/) mice. The s.c. growth of virus-infected Meth A cells in the mutant mice was markedly inhibited by s.c. injections of IFN-gamma, and the rejection from IFN-gamma(+/+) mice was delayed after specific elimination of macrophages by intravenous (i.v.) injections of dichloromethylene diphosphonatecontaining liposomes. These results suggest an essential role of IFN-gamma and involvement of macrophage in the rejection of CTL-resistant, virus-infected cells.


Calorimetric and potentiometric studies on the binding of calcium by phytic acid.
Potentiometric as well as thermal titrations of phytic acid and its calcium complexes have been conducted using both the batch and titration microcalorimeters. For phytic acid, the experimental values by either method are in excellent agreement. For the calcium complexes, the total number of groups and the total heat evolved are in agreement but the placement of the curves is different due primarily to the differences in calcium concentration. Binding of calcium by phytic acid is endothermic for the pH range 2.0-12.0 while the heat of dilution of 1 M CaCl2 is exothermic. The binding at pH greater than 11 gives a value of 22.5 +/- 0.9 kcal mol-1. The same enthalpy of binding (22.3 +/- 0.6 kcal mol-1) could be calculated for the entire pH range studied which requires a knowledge of the observed heat of binding, the thermal titration curves of the acid and its calcium complex, and the change in the hydrogen ion environment. Inspection of the thermal binding curves at pH greater than 11 indicates that a number of step-binding constants are involved and that 5.2 mol calcium are bound/mole phytic acid. This value has been confirmed by atomic absorption spectroscopy. Both the thermal and the potentiometric curves are reversible either by the instantaneous injection of acid or base or by continuous titration. Values for the ionization constants (as pK') and the enthalpy of ionization (as delta Hi) have been estimated by computer assisted curve fitting.


Granulocytic ehrlichiosis in two dogs in Switzerland.
This case report describes two dogs with granulocytic ehrlichiosis. Dog 1 was a male Labrador retriever with clinical signs of lymphosarcoma. Dog 2 was a female Airedale terrier, whose clinical signs included apathy, pyrexia, diarrhea, and abdominal pain. Examination of blood smears revealed Ehrlichia organisms in the neutrophils of both dogs. There was thrombocytopenia in both dogs, and dog 2 also had leukopenia. In both dogs, bands of identical length were amplified from DNA of leukocytes via nested PCR. The bands had identical nucleotide sequences, which differed from the gene sequences of Ehrlichia equi and E. phagocytophila in three and two positions, respectively. Interestingly, the nucleotide sequence of the 16S rRNA was 100% homologous to that of a human granulocytic ehrlichia.


A comparison of the biological properties of European red deer (Cervus elaphus elaphus) spermatozoa stored in the epididymides and in a liquid state at 5 °C.
The biological properties of European red deer (Cervus elaphus elaphus) spermatozoa stored in the epididymides and in a liquid state were compared. Spermatozoa were collected from the epididymides harvested post-mortem. In the first variant, spermatozoa were diluted in two extenders (Bovidyl® and Salomon's), and were stored at 5 °C for up to 144 h. In the second variant, spermatozoa were stored in the epididymides at 5 °C for up to 144 h, and then diluted in the same extenders. Biological properties were evaluated after 0, 48, 96, and 144 h of storage. Sperm motility parameters were determined in the CASA system. Plasma and acrosomal membrane integrity, mitochondrial activity, apoptotic changes, and DNA integrity were assessed by the fluorescence method. Most variables were significantly influenced by the storage method and time. At 144 h, differences (P ≤ 0.05) in sperm parameters were observed between storage variants. Total motility (TMOT), plasma membrane integrity, and mitochondrial activity decreased below 50% of baseline values in the spermatozoa stored in the epididymides, but remained above 70% of baseline values in the spermatozoa stored in a liquid state. The compared storage variants did not differ in TMOT, mitochondrial activity, or the percentage of viable spermatozoa without apoptotic-like changes up to 96 h of storage, regardless of the applied extender. The earliest significant changes were noted in sperm motility parameters. In conclusion, European red deer spermatozoa can be stored in the epididymides at 5 °C for up to 96 h, but their biological parameters are more effectively preserved during liquid storage.


The Global Fund: managing great expectations.
The Global Fund to fight AIDS, Tuberculosis, and Malaria was created to increase funds to combat these three devastating diseases. We report interim findings, based on interviews with 137 national-level respondents that track early implementation processes in four African countries. Country coordinating mechanisms (CCMs) are country-level partnerships, which were formed quickly to develop and submit grant proposals to the Global Fund. CCM members were often ineffective at representing their constituencies and encountered obstacles in participating in CCM processes. Delay in dissemination of guidelines from the Global Fund led to uncertainty among members about the function of these new partnerships. Respondents expressed most concern about the limited capacity of fund recipients--government and non-government--to meet Global Fund conditions for performance-based disbursement. Delays in payment of funds to implementing agencies have frustrated rapid financing of disease control interventions. The Global Fund is one of several new global initiatives superimposed on existing country systems to finance the control of HIV/AIDS. New and existing donors need to coordinate assistance to developing countries by bringing together funding, planning, management, and reporting systems if global goals for disease control are to be achieved.


A multisample study of longitudinal changes in brain network architecture in 4-13-year-old children.
Recent advances in human neuroimaging research have revealed that white-matter connectivity can be described in terms of an integrated network, which is the basis of the human connectome. However, the developmental changes of this connectome in childhood are not well understood. This study made use of two independent longitudinal diffusion-weighted imaging data sets to characterize developmental changes in the connectome by estimating age-related changes in fractional anisotropy (FA) for reconstructed fibers (edges) between 68 cortical regions. The first sample included 237 diffusion-weighted scans of 146 typically developing children (4-13 years old, 74 females) derived from the Pediatric Longitudinal Imaging, Neurocognition, and Genetics (PLING) study. The second sample included 141 scans of 97 individuals (8-13 years old, 62 females) derived from the BrainTime project. In both data sets, we compared edges that had the most substantial age-related change in FA to edges that showed little change in FA. This allowed us to investigate if developmental changes in white matter reorganize network topology. We observed substantial increases in edges connecting peripheral and a set of highly connected hub regions, referred to as the rich club. Together with the observed topological differences between regions connecting to edges showing the smallest and largest changes in FA, this indicates that changes in white matter affect network organization, such that highly connected regions become even more strongly imbedded in the network. These findings suggest that an important process in brain development involves organizing patterns of inter-regional interactions. Hum Brain Mapp 39:157-170, 2018. © 2017 Wiley Periodicals, Inc.


RapidChek SELECT Salmonella. Performance Tested Method 080601.
RapidChek SELECT Salmonella was previously validated in the Performance Tested Methods program for the detection of Salmonella spp. in raw ground chicken, chicken carcass rinse, sliced cooked turkey, and liquid eggs. The present matrix extension study conducted under the AOAC Research Institute Emergency Response Validation program compared the RapidChek SELECT Salmonella method to the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA-BAM) method for the detection of Salmonella Typhimurium in peanut butter. Overall, 27 samples were found positive by the RapidChek SELECT Salmonella method and 27 were found to be positive by the reference method. All RapidChek SELECT Salmonella presumptive positives were confirmed positive by the cultural reference method; additionally, all presumptive negative results were confirmed negative by the cultural reference method. Accordingly 0% false-negative rate and 0% false-positive rate were found. No significant difference between the RapidCheck SELECT Salmonella and FDA-BAM reference method was found; calculated Chi-square was 0. Results indicate that a low level of Salmonella in peanut butter can be successfully recovered and detected in the minimum 24 h enrichment protocol.


Ultrahigh resolution spectral analysis based on a Brillouin fiber laser.
We report what we believe to be a novel experimental heterodyne technique for the spectral analysis of continuous optical wave sources. The achieved resolution is as low as the kilohertz level, with a dynamic range in excess of 90 dB. The technique is based on a heterodyne detection between the source under test (SUT) and a Brillouin fiber laser generated by this SUT. Contrary to standard self-heterodyne techniques only a few tens of meters of optical fiber is required without the need of any optical modulators. This spectrometer has been used to characterize a distributed-feedback laser diode and a Brillouin fiber laser.


Antipsychotic drug use and the risk of venous thromboembolism in elderly patients.
Our aim was to investigate the relationship between exposure to antipsychotic drugs and the risk of venous thromboembolism (VTE) in elderly patients. A time-matched case-control analysis nested within a cohort of 111,818 patients with at least 1 antipsychotic drug prescription during 1998 to 2008. Data were used from the PHARMO institute's database, which contains drug dispensing data from community pharmacies and hospital admission data. The index date was for the cases defined as the date of hospital admission for VTE (deep venous thrombosis [DVT] or pulmonary embolism) or, for outpatient cases, the start of therapeutic dose low-molecular weight heparin therapy. For each case, 4 controls matched by age and sex were randomly sampled from the cohort. Two measures were used to evaluate the temporal relationship between antipsychotic drug use and the occurrence of VTE: being a current, recent, or past user and the duration of use up to the index date. The strength of the association was expressed as odds ratios with 95% confidence intervals, taking into account potential confounders. We identified 367 cases of hospital admission for DVT, 342 cases of hospital admission for PE, and 323 cases of outpatient treatment of DVT. Current exposure to antipsychotic drugs was not associated with an increased risk of VTE, compared with nonusers (odds ratio, 0.9; 95% confidence interval, 0.7-1.1). We found no association between dosage, the duration of use, or the type of antipsychotic drug and the risk of VTE. We found no evidence of an increased risk of VTE in elderly patients using antipsychotic drugs.


Escherichia coli aconitases and oxidative stress: post-transcriptional regulation of sodA expression.
Escherichia coli possesses two aconitases, a stationary-phase enzyme (AcnA), which is induced by iron and oxidative stress, and a major but less stable enzyme (AcnB), synthesized during exponential growth. In addition to the catalytic activities of the holo-proteins, the apo-proteins function as post-transcriptional regulators by site-specific binding to acn mRNAs. Thus, it has been suggested that inactivation of the enzymes could mediate a rapidly reacting post-transcriptional component of the bacterial oxidative stress response. Here it is shown that E. coli acn mutants are hypersensitive to the redox-stress reagents H(2)O(2) and methyl viologen. Proteomic analyses further revealed that the level of superoxide dismutase (SodA) is enhanced in acnB and acnAB mutants, and by exposure to methyl viologen. The amounts of other proteins, including thioredoxin reductase, 2-oxoglutarate dehydrogenase, succinyl-CoA synthetase and chaperone proteins, were also affected in the acn mutants. The altered patterns of sodA expression were confirmed in studies with sodA-lacZ reporter strains. Quantitative Northern blotting indicated that AcnA enhances the stability of the sodA transcript, whereas AcnB lowers its stability. Direct evidence that the apo-proteins have positive (AcnA) and negative (AcnB) effects on SodA synthesis was obtained from in vitro transcription-translation experiments. It is suggested that the aconitase proteins of E. coli serve as a protective buffer against the basal level of oxidative stress that accompanies aerobic growth by acting as a sink for reactive oxygen species and by modulating translation of the sodA transcript.


Role of arterial stiffness in the association between hand grip strength and cardiovascular events: the Korean Genome and Epidemiology Study.
Hand grip strength (HGS) has been associated with cardiovascular events. However, the exact mechanism responsible for the inverse association between HGS and cardiovascular events has not been established. The aim of this study was to assess whether arterial stiffness mediates this association. We studied 1508 participants (age; 60 ± 5, men; 47.5%) from the Ansan cohort of the Korean Genome Epidemiology Study. Participants were assessed for various parameters of arterial stiffness as well as HGS. The augmentation index (AIx) and brachial-ankle pulse wave velocity (baPWV) were evaluated by using an applanation tonometer and automated waveform analyzer, respectively. Carotid intima medial thickness (IMT) was measured by B-mode ultrasonogram with a 7.5-MHz linear array transducer. HGS was evaluated using a Jamar dynamometer. With increased grip strength, AIx decreased (r = 0.437, P < 0.001). baPWV (r = 0.044, P = 0.107) and carotid IMT (r = 0.005, P = 0.856) had no significant correlation with grip strength. This trend was consistently observed regardless of hypertension, but was more pronounced in participants with hypertension. HGS was significantly correlated with AIx, but not with baPWV and carotid IMT. Our findings suggest that central arterial stiffness could mediate the association between HGS and cardiovascular events.


Endogenous hormones and coronary heart disease in postmenopausal women.
The association between serum levels of endogenous estrogens in postmenopausal women and the subsequent risk of coronary heart disease (CHD) was examined in a prospective case-control study nested within the New York University Women's Health Study (NYUWHS). The NYUWHS is a prospective cohort study of 14,274 healthy women enrolled between 1985 and 1991. A total of 99 women who were postmenopausal and free of cardiovascular disease at enrollment and who subsequently experienced CHD, defined as non-fatal myocardial infarction (MI), fatal CHD, percutaneous transluminal coronary angioplasty (PTCA), or coronary artery bypass grafting (CABG), were matched 1:2 by baseline age, blood sampling date, and postmenopausal status to controls who remained free of CHD as of the date of diagnosis of the matching case. Biochemical analyses for total estradiol, estrone, percent free estradiol, percent estradiol bound to sex hormone-binding globulin (SHBG), and SHBG were performed on pre-diagnostic stored serum samples. Participants had not used any hormone medications in the 6 months prior to blood collection. In the model adjusting only for matching factors, the risk of CHD in the top tertile of calculated bioavailable estradiol was elevated compared with the bottom tertile (OR=2.10; 95% CI=1.13-3.90, P for trend=0.03), and the risk in the top tertile of SHBG was reduced (OR=0.50, 95% CI=0.28-0.92, P for trend<0.01). However, these associations disappeared after adjusting for baseline hypertension status, body mass index, and serum cholesterol levels. These findings suggest that circulating estradiol and SHBG are not associated with CHD risk in postmenopausal women beyond what can be explained by the variation in hypertension status, BMI, and cholesterol.


Altered expression of calcium-activated K and Cl channels in detrusor overactivity of rats with partial bladder outlet obstruction.
To evaluate the activity of large- and small-conductance calcium-activated potassium channels (BKCa, SKCa) and calcium-activated chloride channels (ClCa) in detrusor overactivity (DO) cells after partial bladder outlet obstruction (PBOO) in rats. Thirteen female Wistar rats with DO caused by PBOO were studied simultaneously with eight sham-operated rats. The expression of KCa and ClCa channels was assessed by reverse transcription-polymerase chain reaction, and the function of the two groups compared. In the DO cells the expression of BKCa, SKCa2 and SKCa3 was lower, and that of ClCa channels higher, than in the control group cells. Using confocal laser scanning microscopic analysis, the function of BKCa and SKCa channels was suppressed, and that of ClCa channels was enhanced in DO group cells. KCa and ClCa effectors altered the cell membrane potentials more significantly in the DO cells than in the control cells, indicating a decrease in KCa and an increase in ClCa in DO group in either iso- or hypo-osmolar medium. Moreover, the change in BKCa, SKCa and ClCa channel activators in DO cells showed a more excitable state in hypo-osmolar medium than in iso-osmolar medium. In DO myocytes after PBOO, the expression and function of KCa channels were decreased, and those of ClCa channels increased. These changes all provoke greater cell excitability, and could partly account for the DO.


Differential grazing of two heterotrophic nanoflagellates on marine Synechococcus strains.
Grazing of heterotrophic nanoflagellates on marine picophytoplankton presents a major mortality factor for this important group of primary producers. However, little is known of the selectivity of the grazing process, often merely being thought of as a general feature of cell size and motility. In this study, we tested grazing of two heterotrophic nanoflagellates, Paraphysomonas imperforata and Pteridomonas danica, on strains of marine Synechococcus. Both nanoflagellates proved to be selective in their grazing, with Paraphysomonas being able to grow on 5, and Pteridomonas on 11, of 37 Synechococcus strains tested. Additionally, a number of strains (11 for Paraphysomonas, 9 for Pteridomonas) were shown to be ingested, but not digested (and thus did not support growth of the grazer). Both the range of prey strains that supported growth as well as those that were ingested but not digested was very similar for the two grazers, suggesting a common property of these prey strains that lent them susceptible to grazing. Subsequent experiments on selected Synechococcus strains showed a pronounced difference in grazing susceptibility between wild-type Synechococcus sp. WH7803 and a spontaneous phage-resistant mutant derivative, WH7803PHR, suggesting that cell surface properties of the Synechococcus prey are an important attribute influencing grazing vulnerability.


Streptomyces thermocoprophilus sp. nov., a cellulase-free endo-xylanase-producing streptomycete.
The taxonomic position of a thermophilic actinomycete strain isolated from poultry faeces was examined using a polyphasic approach. The isolate, designated B19T, was assigned to the genus Streptomyces on the basis of chemotaxonomic and morphological criteria. An almost complete 16S rRNA gene (rDNA) sequence obtained for the test strain was compared with those of representative streptomycetes, notably thermophilic streptomycetes. 16S rDNA sequence data not only supported the assignment of the strain to the genus Streptomyces but also showed that the isolate formed a distinct phyletic line within the evolutionary branch composed of Streptomyces thermodiastaticus and related species. The strain was distinguished from related validly described Streptomyces species by a number of phenotypic properties. It is, therefore, proposed that strain B19T be classified in the genus Streptomyces as Streptomyces thermocoprophilus sp. nov.


Preventing (impulsive) errors: Electrophysiological evidence for online inhibitory control over incorrect responses.
In a rich environment, with multiple action affordances, selective action inhibition is critical in preventing the execution of inappropriate responses. Here, we studied the origin and the dynamics of incorrect response inhibition and how it can be modulated by task demands. We used EEG in a conflict task where the probability of compatible and incompatible trials was varied. This allowed us to modulate the strength of the prepotent response, and hence to increase the risk of errors, while keeping the probability of the two responses equal. The correct response activation and execution was not affected by compatibility or by probability. In contrast, incorrect response inhibition in the primary motor cortex ipsilateral to the correct response was more pronounced on incompatible trials, especially in the condition where most of the trials were compatible, indicating a modulation of inhibitory strength within the course of the action. Two prefrontal activities, one medial and one lateral, were also observed before the response, and their potential links with the observed inhibitory pattern observed are discussed.


From promising molecules to orphan drugs: Early clinical drug development.
Phase-1 (also known as "First-in-Man") clinical trials initiate the early clinical development of possible new medicines. Patient participation in this early phase of clinical trials is rather limited. After successful phase 1 trials, further phase 2 and phase 3 clinical trials in patients may lead to a marketing authorization. In the first 15 years of the European Union Orphan Drug Directive, 4.5% of the orphan drug applications were authorized. However, for many of these orphan drugs, no phase 1 studies were required, as these products were already well known pharmaceutical substances, with a clearly defined pharmacological profile. Furthermore, for 19 orphan drugs, already authorized by the European Medicines Agency (EMA), the original rare indication was extended to another rare disease and no phase 1 trials were needed. Phase 1 studies need to be performed in a sufficient number of volunteers even for medicinal products intended for a very limited number of patients.


Clinical outcomes of tracheoesophageal diversion for intractable aspiration.
The purpose of the present study was to examine the clinical outcomes of using tracheoesophageal diversion for preventing intractable aspiration. We retrospectively reviewed 25 patients who underwent tracheoesophageal diversion from 2003 to 2009 at our hospital (median age, 25 years; range, 0-78 years). End-to-side anastomosis was used in 16 cases and side-to-side anastomosis was used in 9. The average operative time was 141 minutes for end-to-side anastomosis and 191 minutes for side-to-side anastomosis. Peri-operative complications were observed in only two (8 per cent) cases: one with infection and one with haematoma. No fistulas were observed. Aspiration was prevented in all cases, but the nutritional route depended on the swallowing function of the patient. Oral feeding was the main nutritional route after surgery in only four patients (16 per cent). This procedure is well suited to patients who lack speech communication and are at high risk of aspiration.


The impact of hypertension on retinal nerve fiber layer thickness and its association with carotid intima media thickness.
Our aim was to investigate retinal nerve fiber layer (RNFL) thickness in hypertensive patients using spectral-domain optical coherence tomography (SD-OCT) and to evaluate the relationship between RNFL thickness and carotid intima media thickness (CIMT). This study included 59 patients with hypertension (HT) (53.6 ± 10.7 years) and 54 age-matched healthy controls (51.0 ± 8.1 years). We evaluated anthropometric and metabolic parameters as well as RNFL and CIMT measurements in patients with hypertension and controls. The average RNFL thickness was 86.60 ± 10.86 μm in hypertensive patients and 93.63 ± 7.30 μm in healthy controls (p < 0.001). Selective thinning of the RNFL was found in the superior and inferior quadrants. Mean CIMT values were higher in patients with HT (0.80 ± 0.15 mm) than the healthy subjects (0.71 ± 0.1 mm) (p < 0.001). The average, inferior and nasal RNFL thickness were negatively associated with diastolic blood pressure respectively (r = - 0.112, r = - 0.210, r = - 0.225). There was an inverse correlation between RNFL thickness in the average and superior retinal quadrant and CIMT (r = - 0.201, r = - 0.185). There were no correlations between RNFL thickness and age, body mass index, fasting plasma glucose, lipid parameters, high-sensitive C-reactive protein and microalbuminuria. RNFL thickness is reduced in hypertensive patients and may be associated with atherosclerosis.


Synaptic vesicle dynamics in rat fast and slow motor nerve terminals.
We have investigated whether rat motor nerve terminals with different in vivo activity patterns also have different vesicle trafficking characteristics. To do this, we monitored, using combined optical and electrical techniques, the rate of exocytosis (during different frequencies and patterns of activity), the releasable pool size, and the recycle time of synaptic vesicles in terminals on soleus (slow-twitch) and extensor digitorum longus [(EDL); fast-twitch] muscle fibers. EDL terminals had a higher initial quantal content (QC) than soleus, but during tonic or phasic stimulation at 20-80 Hz, EDL QC ran down to a greater extent than soleus QC. By recording loss of fluorescence from exocytosing vesicles labeled with the dye FM1-43, EDL terminals were found to destain faster than those in soleus. Simultaneous intracellular recording of end plate potentials, to count the number of vesicles released, permitted estimation of the total vesicle pool (VP) size and the recycle time by combining the optical and electrophysiological data. Soleus vesicle pool was larger than EDL, but recycle time was not significantly different. These terminals, therefore, are adapted to their in vivo activity patterns by alterations in QC and VP size but not recycle time.


Adenosine A2A Receptor Gene Knockout Prevents l-3,4-Dihydroxyphenylalanine-Induced Dyskinesia by Downregulation of Striatal GAD67 in 6-OHDA-Lesioned Parkinson's Mice.
l-3,4-Dihydroxyphenylalanine (l-DOPA) remains the primary pharmacological agent for the symptomatic treatment of Parkinson's disease (PD). However, the development of l-DOPA-induced dyskinesia (LID) limits the long-term use of l-DOPA for PD patients. Some data have reported that adenosine A2A receptor (A2AR) antagonists prevented LID in animal model of PD. However, the mechanism in which adenosine A2AR blockade alleviates the symptoms of LID has not been fully clarified. Here, we determined to knock out (KO) the gene of A2AR and explored the possible underlying mechanisms implicated in development of LID in a mouse model of PD. A2AR gene KO mice were unilaterally injected into the striatum with 6-hydroxydopamine (6-OHDA) in order to damage dopamine neurons on one side of the brain. 6-OHDA-lesioned mice were then injected once daily for 21 days with l-DOPA. Abnormal involuntary movements (AIMs) were evaluated on days 3, 8, 13, and 18 after l-DOPA administration, and real-time polymerase chain reaction and immunohistochemistry for glutamic acid decarboxylase (GAD) 65 and GAD67 were performed. We found that A2AR gene KO was effective in reducing AIM scores and accompanied with decrease of striatal GAD67, rather than GAD65. These results demonstrated that the possible mechanism involved in alleviation of AIM symptoms by A2AR gene KO might be through reducing the expression of striatal GAD67.


Studies toward the total synthesis of clavulactone.
Synthetic studies directed toward a total synthesis of clavulactone are reported. In light of the analysis made in our previous work, cyclopentane 4a (a key intermediate in the present work) was synthesized through a radical-mediated ring closure of a rationally designed substrate 25. Using HWE reactions, the lower and upper side-chains of 4a were converted into an allyl chloride and an allyl cyanohydrin, respectively. Subsequent treatment of the allyl chloride/cyanohydrin in a highly diluted THF solution with sodium bis(trimethylsiliyl)amide led to intramolecular alkylation and thus completed a major endeavor in synthesizing the dolabellane framework, construction of the eleven-membered ring. SmI(2)-mediated lactonization as a model reaction for the formation of the alpha,beta-unsaturated delta-lactone segment of clavulactone is also described.


Effectiveness Analysis of a Part-Time Rapid Response System During Operation Versus Nonoperation.
To evaluate the effect of a part-time rapid response system on the occurrence rate of cardiopulmonary arrest by comparing the times of rapid response system operation versus nonoperation. Retrospective cohort study. A 1,360-bed tertiary care hospital. Adult patients admitted to the general ward were screened. Data were collected over 36 months from rapid response system implementation (October 2012 to September 2015) and more than 45 months before rapid response system implementation (January 2009 to September 2012). None. The rapid response system operates from 7 AM to 10 PM on weekdays and from 7 AM to 12 PM on Saturdays. Primary outcomes were the difference of cardiopulmonary arrest incidence between pre-rapid response system and post-rapid response system periods and whether the rapid response system operating time affects the cardiopulmonary arrest incidence. The overall cardiopulmonary arrest incidence (per 1,000 admissions) was 1.43. Although the number of admissions per month and case-mix index were increased (3,555.18 vs 4,564.72, p < 0.001; 1.09 vs 1.13, p = 0.001, respectively), the cardiopulmonary arrest incidence was significantly decreased after rapid response system (1.60 vs 1.23; p = 0.021), and mortality (%) was unchanged (1.38 vs 1.33; p = 0.322). After rapid response system implementation, the cardiopulmonary arrest incidence significantly decreased by 40% during rapid response system operating times (0.82 vs 0.49/1,000 admissions; p = 0.001) but remained similar during rapid response system nonoperating times (0.77 vs 0.73/1,000 admissions; p = 0.729). The implementation of a part-time rapid response system reduced the cardiopulmonary arrest incidence based on the reduction of cardiopulmonary arrest during rapid response system operating times. Further analysis of the cost effectiveness of part-time rapid response system is needed.


An Outcome Study of Anterior Cervical Discectomy and Fusion among Iranian Population.
Background and Aim. First-line treatment strategy for managing cervical disc herniation is conservative measures. In some cases, surgery is indicated either due to signs/symptoms of severe/progressive neurological deficits, or because of persistence of radicular pain despite 12 weeks of conservative treatment. Success for treatment of cervical disc herniation using ACDF has been successfully reported in the literature. We aim to determine the outcome of ACDF in treatment of cervical disc herniation among Iranians. Methods and Materials/Patients. In a retrospective cohort study, we evaluated 68 patients who had undergone ACDF for cervical disc herniation from March 2006 to March 2011. Outcome tools were as follows: (1) study-designed questionnaire that addressed residual and/or new complaints and subjective satisfaction with the operation; (2) recent (one week prior to the interview) postoperative VAS for neck and upper extremity radicular pain; (3) Japanese Orthopaedic Association Myelopathy Evaluation Questionnaire (JOACMEQ) (standard Persian version); and (4) follow-up cervical Magnetic Resonance Imaging (MRI) and lateral X-ray. Results. With mean follow-up time of 52.93 (months) ± 31.89 SD (range: 13-131 months), we had success rates with regard to ΔVAS for neck and radicular pain of 88.2% and 89.7%, respectively. Except QOL functional score of JOAMEQ, 100% success rate for the other 4 functional scores of JOAMEQ was achieved. Conclusion. ACDF is a successful surgical technique for the management of cervical disc herniation among Iranian population.


Slow Inactivation of Sodium Channels Contributes to Short-Term Adaptation in Vomeronasal Sensory Neurons.
Adaptation plays an important role in sensory systems as it dynamically modifies sensitivity to allow the detection of stimulus changes. The vomeronasal system controls many social behaviors in most mammals by detecting pheromones released by conspecifics. Stimuli activate a transduction cascade in vomeronasal neurons that leads to spiking activity. Whether and how these neurons adapt to stimuli is still debated and largely unknown. Here, we measured short-term adaptation performing current-clamp whole-cell recordings by using diluted urine as a stimulus, as it contains many pheromones. We measured spike frequency adaptation in response to repeated identical stimuli of 2-10 s duration that was dependent on the time interval between stimuli. Responses to paired current steps, bypassing the signal transduction cascade, also showed spike frequency adaptation. We found that voltage-gated Na+ channels in VSNs undergo slow inactivation processes. Furthermore, recovery from slow inactivation of voltage-gated Na+ channels occurs in several seconds, a time scale similar to that measured during paired-pulse adaptation protocols, suggesting that it partially contributes to short-term spike frequency adaptation. We conclude that vomeronasal neurons do exhibit a time-dependent short-term spike frequency adaptation to repeated natural stimuli and that slow inactivation of Na+ channels contributes to this form of adaptation. These findings not only increase our knowledge about adaptation in the vomeronasal system, but also raise the question of whether slow inactivation of Na+ channels may play a role in other sensory systems.


CD63 associates with tyrosine kinase activity and CD11/CD18, and transmits an activation signal in neutrophils.
As a member of the tetraspan family, it has been hypothesized that CD63 may be associated with signal transduction; however, its role in leukocyte function is unknown. To examine the potential ability of CD63 to activate neutrophils, the effects of five CD63 mAbs, AHN-16, -16.1, -16.2, -16.3, and -16.5, were examined for their ability to alter neutrophil adhesion to HUVEC monolayers. These CD63 Abs increased neutrophil adhesion to resting and TNF-stimulated HUVEC monolayers. This increase in neutrophil adhesion caused by CD63 Abs was blocked by a CD18 Ab and was associated with up-regulation of CD11/CD18 and down-regulation of CD62L on the neutrophil surface. CD11/CD18 was also found to be associated with CD63. This increase in neutrophil adhesion required physiologic extracellular calcium concentrations at or near the time of CD63 Ab binding. The incubation of CD63 Abs with cells in the absence of calcium for 10 min before repletion of calcium resulted in no increase in neutrophil adhesion. Protein kinase activity was detected in neutrophils associated with CD63. Most of the protein kinase activity associated with these Ags was tyrosine kinase activity, with a lesser amount of threonine and serine kinase activities. Src family kinases Lyn and Hck accounted for much of the associated tyrosine kinase activity. The data suggest that CD63 Ab binding to the neutrophil surface triggers a transient activation signal that requires extracellular calcium and regulates the adhesive activity of CD11/CD18. Associated protein kinase activity may play a role in signal transduction by CD63 to regulate other cell functions.


Comparative genomics of Lactobacillus.
The genus Lactobacillus includes a diverse group of bacteria consisting of many species that are associated with fermentations of plants, meat or milk. In addition, various lactobacilli are natural inhabitants of the intestinal tract of humans and other animals. Finally, several Lactobacillus strains are marketed as probiotics as their consumption can confer a health benefit to host. Presently, 154 Lactobacillus species are known and a growing fraction of these are subject to draft genome sequencing. However, complete genome sequences are needed to provide a platform for detailed genomic comparisons. Therefore, we selected a total of 20 genomes of various Lactobacillus strains for which complete genomic sequences have been reported. These genomes had sizes varying from 1.8 to 3.3 Mb and other characteristic features, such as G+C content that ranged from 33% to 51%. The Lactobacillus pan genome was found to consist of approximately 14 000 protein-encoding genes while all 20 genomes shared a total of 383 sets of orthologous genes that defined the Lactobacillus core genome (LCG). Based on advanced phylogeny of the proteins encoded by this LCG, we grouped the 20 strains into three main groups and defined core group genes present in all genomes of a single group, signature group genes shared in all genomes of one group but absent in all other Lactobacillus genomes, and Group-specific ORFans present in core group genes of one group and absent in all other complete genomes. The latter are of specific value in defining the different groups of genomes. The study provides a platform for present individual comparisons as well as future analysis of new Lactobacillus genomes.


Synchronization patterns: from network motifs to hierarchical networks.
We investigate complex synchronization patterns such as cluster synchronization and partial amplitude death in networks of coupled Stuart-Landau oscillators with fractal connectivities. The study of fractal or self-similar topology is motivated by the network of neurons in the brain. This fractal property is well represented in hierarchical networks, for which we present three different models. In addition, we introduce an analytical eigensolution method and provide a comprehensive picture of the interplay of network topology and the corresponding network dynamics, thus allowing us to predict the dynamics of arbitrarily large hierarchical networks simply by analysing small network motifs. We also show that oscillation death can be induced in these networks, even if the coupling is symmetric, contrary to previous understanding of oscillation death. Our results show that there is a direct correlation between topology and dynamics: hierarchical networks exhibit the corresponding hierarchical dynamics. This helps bridge the gap between mesoscale motifs and macroscopic networks.This article is part of the themed issue 'Horizons of cybernetical physics'.


Thalamic deep brain stimulation for writer's cramp.
Writer's cramp is a type of idiopathic focal hand dystonia characterized by muscle cramps that accompany execution of the writing task specifically. In this report, the authors describe the clinical outcome after thalamic deep brain stimulation (DBS) therapy in patients with writer's cramp and present an illustrative case with which they compare the effects of pallidal and thalamic stimulation. In addition to these results for the clinical effectiveness, they also examine the best point and pattern for therapeutic stimulation of the motor thalamus, including the nucleus ventrooralis (VO) and the ventralis intermedius nucleus (VIM), for writer's cramp. The authors applied thalamic DBS in five patients with writer's cramp. The inclusion criteria for the DBS trial in this disorder were a diagnosis of idiopathic writer's cramp and the absence of a positive response to medication. The exclusion criteria included significant cognitive dysfunction, active psychiatric symptoms, and evidence of other central nervous system diseases or other medical disorders. In one of the cases, DBS leads were implanted into both the globus pallidus internus and the VO/VIM, and test stimulation was performed for 1 week. The authors thus had an opportunity to compare the effects of pallidal and thalamic stimulation in this patient. Immediately after the initiation of thalamic stimulation, the neurological deficits associated with writer's cramp were improved in all five cases. Postoperatively all preoperative scale scores indicating the seriousness of the writer's cramp were significantly lower (p < 0.001). In the patient in whom two DBS leads were implanted, the clinical effect of thalamic stimulation was better than that of pallidal stimulation. During the thalamic stimulation, the maximum effect was obtained when stimulation was applied to both the VO and the VIM widely, compared with being applied only within the VO. The authors successfully treated patients with writer's cramp by thalamic DBS. Insofar as they are aware, this is the first series in which writer's cramp has been treated with DBS. Thalamic stimulation appears to be a safe and valuable therapeutic option for writer's cramp.


Predicting Remission in Subjects at Clinical High Risk for Psychosis Using Mismatch Negativity.
The declining transition rate to psychotic disorder and the increasing rate of nonpsychotic poor outcomes among subjects at clinical high risk (CHR) for psychosis have increased the need for biomarkers to predict remission regardless of transition. This study investigated whether mismatch negativity (MMN) predicts the prognosis of CHR individuals during a 6-year follow-up period. A total of 47 healthy control (HC) subjects and 48 subjects at CHR for psychosis participated in the MMN assessment. The clinical statuses of the CHR subjects were examined at baseline and regularly for up to 6 years. The CHR subjects were divided into remitter and nonremitter groups, and the baseline MMN amplitudes and latencies were compared across the remitter, nonremitter, and HC groups. Regression analyses were performed to identify the predictive factors of remission, the improvement of attenuated positive symptoms, and functional recovery. CHR nonremitters showed reduced MMN amplitudes at baseline compared to CHR remitters and HC subjects. A logistic regression analysis revealed that the baseline MMN amplitude at the frontal electrode site was the only significant predictor of remission. In a multiple regression analysis, the MMN amplitude, antipsychotic use, and years of education predicted an improvement in attenuated positive symptoms. The MMN amplitude at baseline predicted functional recovery. These results suggest that MMN is a putative predictor of prognosis regardless of the transition to psychotic disorder in subjects at CHR. Early prognosis prediction and the provision of appropriate interventions based on the initial CHR status might be aided using MMN.


A new pathway for heavy metal detoxification in animals. Phytochelatin synthase is required for cadmium tolerance in Caenorhabditis elegans.
Increasing emissions of heavy metals such as cadmium, mercury, and arsenic into the environment pose an acute problem for all organisms. Considerations of the biochemical basis of heavy metal detoxification in animals have focused exclusively on two classes of peptides, the thiol tripeptide, glutathione (GSH, gamma-Glu-Cys-Gly), and a diverse family of cysteine-rich low molecular weight proteins, the metallothioneins. Plants and some fungi, however, not only deploy GSH and metallothioneins for metal detoxification but also synthesize another class of heavy metal binding peptides termed phytochelatins (PCs) from GSH. Here we show that PC-mediated heavy metal detoxification is not restricted to plants and some fungi but extends to animals by demonstrating that the ce-pcs-1 gene of the nematode worm Caenorhabditis elegans encodes a functional PC synthase whose activity is critical for heavy metal tolerance in the intact organism.


Behavioral, endocrine, immune, and performance measures for pigs exposed to acute stress.
Weanling pigs (n = 132) were used to investigate the effects of three common stressors (and a control) and differing social status on behavior, immunity, plasma cortisol, blood chemical, and performance measures. Eleven blocks of 12 pigs each were evaluated. Each pen contained three pigs of dominant (DOM), intermediate (INT), or submissive (SUB) social status. Two weeks later, random pens of pigs experienced either a control treatment (CON) or they were stressed for 4 h by shipping (SHIP), heat-stressed (HEAT) with overhead heat lamps in their home pens, or cold-stressed (COLD) by direct application of water and an air current. Treatments did not influence body weights; however, percentage weight loss during SHIP was greater than for other treatments. Body weights were heavier for DOM pigs than for INT and SUB pigs. Social status had large effects on plasma cortisol, globulin, acute-phase proteins, body weight, and weight changes. Only acute shipping stress resulted in weight loss. Many immune and blood measures were not changed among acutely stressed pigs; however, the relationship between social status and mitogen-induced lymphocyte proliferation and natural killer cell cytotoxicity was disrupted during acute stress. Pig behavior was significantly changed by each stress treatment in a unique manner. During acute stress, behavioral changes seem to be the most consistent and reliable indicators.


A ribosome dissociation factor from rabbit reticulocytes (fluoride-E. coli ribosomes-initiation factors).
A ribosome dissociation factor has been detected in an extract of ribosomal particles from rabbit reticulocytes. This factor dissociates free ribosomes from reticulocytes and also from Escherichia coli; it does not dissociate ribosomes complexed with peptidyl-tRNA and mRNA. The reaction appears to be stoichiometric rather than catalytic; it reaches completion in one minute at 37 degrees C, but is very slow at 0 degrees C, and it is antagonized and reversed by Mg(++). Reticulocyte dissociation factor thus closely resembles that from E. coli. However, the activity has been found primarily associated with the native large subunits rather than the small subunits in lysates.


Activation of protein kinase C-α and Src kinase increases urea transporter A1 α-2, 6 sialylation.
The urea transporter A1 (UT-A1) is a glycosylated protein with two glycoforms: 117 and 97 kD. In diabetes, the increased abundance of the heavily glycosylated 117-kD UT-A1 corresponds to an increase of kidney tubule urea permeability. We previously reported that diabetes not only causes an increase of UT-A1 protein abundance but also, results in UT-A1 glycan changes, including an increase of sialic acid content. Because activation of the diacylglycerol (DAG)-protein kinase C (PKC) pathway is elevated in diabetes and PKC-α regulates UT-A1 urea transport activity, we explored the role of PKC in UT-A1 glycan sialylation. We found that activation of PKC specifically promotes UT-A1 glycan sialylation in both UT-A1-MDCK cells and rat kidney inner medullary collecting duct suspensions, and inhibition of PKC activity blocks high glucose-induced UT-A1 sialylation. Overexpression of PKC-α promoted UT-A1 sialylation and membrane surface expression. Conversely, PKC-α-deficient mice had significantly less sialylated UT-A1 compared with wild-type mice. Furthermore, the effect of PKC-α-induced UT-A1 sialylation was mainly mediated by Src kinase but not Raf-1 kinase. Functionally, increased UT-A1 sialylation corresponded with enhanced urea transport activity. Thus, our results reveal a novel mechanism by which PKC regulates UT-A1 function by increasing glycan sialylation through Src kinase pathways, which may have an important role in preventing the osmotic diuresis caused by glucosuria under diabetic conditions.


In vivo composition of NMDA receptor signaling complexes differs between membrane subdomains and is modulated by PSD-95 and PSD-93.
Lipid rafts are dynamic membrane microdomains enriched in cholesterol and sphingolipids involved in the compartmentalization of signaling pathways, trafficking and sorting of proteins. At synapses, the glutamatergic NMDA receptor and its cytoplasmic scaffold protein PSD-95 move between postsynaptic density (PSD) and rafts following learning or ischemia. However it is not known whether the signaling complexes formed by these proteins are different in rafts nor the molecular mechanisms that govern their localization. To examine these issues in vivo we used mice carrying genetically encoded tags for purification of protein complexes and specific mutations in NMDA receptors, PSD-95 and other postsynaptic scaffold proteins. Isolation of PSD-95 complexes from mice carrying tandem affinity purification tags showed differential composition in lipid rafts, postsynaptic density and detergent-soluble fractions. Raft PSD-95 complexes showed less CaMKIIalpha and SynGAP and enrichment in Src and Arc/Arg3.1 compared with PSD complexes. Mice carrying knock-outs of PSD-95 or PSD-93 show a key role for PSD-95 in localizing NR2A-containing NMDA receptor complexes to rafts. Deletion of the NR2A C terminus or the C-terminal valine residue of NR2B, which prevents all PDZ interactions, reduced the NR1 association with rafts. Interestingly, the deletion of the NR2B valine residue increased the total amount of lipid rafts. These data show critical roles for scaffold proteins and their interactions with NMDA receptor subunits in organizing the differential expression in rafts and postsynaptic densities of synaptic signaling complexes.


Long-lived polarization protected by symmetry.
In this paper we elucidate, theoretically and experimentally, molecular motifs which permit Long-Lived Polarization Protected by Symmetry (LOLIPOPS). The basic assembly principle starts from a pair of chemically equivalent nuclei supporting a long-lived singlet state and is completed by coupling to additional pairs of spins. LOLIPOPS can be created in various sizes; here we review four-spin systems, introduce a group theory analysis of six-spin systems, and explore eight-spin systems by simulation. The focus is on AA'XnX'n spin systems, where typically the A spins are (15)N or (13)C and X spins are protons. We describe the symmetry of the accessed states, we detail the pulse sequences used to access these states, we quantify the fraction of polarization that can be stored as LOLIPOPS, we elucidate how to access the protected states from A or from X polarization and we examine the behavior of these spin systems upon introduction of a small chemical shift difference.


Elevating physical activity as a public health priority: creation of the National Society of Physical Activity Practitioners in Public Health.
Physical activity has emerged as a distinct area of public health practice. As this field evolved, the need for a professional organization for physical activity practitioners in public health became evident. A collaboration of several existing public health professional organizations formed to address this new area of public health practice. The collaboration laid the foundation to establish a professional organization. National Association of Physical Activity Practitioners in Public Health (NSPAPPH) was launched in April 2006. NSPAPPH accomplishments to date include convening a national meeting of physical activity practitioners, conducting strategic planning, adopting bylaws and core competencies for professional practice, developing a website and electronic newsletter, and establishing training opportunities for practitioners. Future plans for NSPAPPH include development of a professional certification for physical activity practitioners in public health; enhancement of training and professional development opportunities; recruitment of members from national, tribal, state, and local organizations working in public and private sectors; publications of journal articles, reports, and issue briefs; and development of a policy agenda. Implementing these plans will serve to strengthen public health infrastructure for physical activity, thus improving the physical activity behaviors of Americans and the health of the nation.


When robots fail: The complex processes of learning and development.
Although robots can contribute to the understanding of biological behavior, they fail to model the processes by which humans cope with their environment. Both development and learning are characterized by complex relationships that require constant modification. Given present technology, robots can only model behaviors in specific situations and during discrete stages. Robots cannot master the complex relationships that are the hallmark of human behavior.


Proctographic findings and symptoms in patients with anterior rectoanal intussusception.
Rectoanal intussusception (RAI) is a common finding on defecography in patients with defecation disorders. This study aimed to compare the proctographic findings and symptoms between patients with anterior RAI and those with circular RAI. We included 208 patients who were diagnosed as having RAI on defecography. Anorectal function was evaluated using Constipation Scoring System (CSS) and Fecal Incontinence Severity Index (FISI). Twenty-four patients had anterior RAI and 184 had circular RAI. While the anterior intussusception descent or pelvic floor descent was significantly smaller in patients with anterior RAI than those with circular RAI [14.3 vs. 18.5 mm, p=0.004; 12.4 vs. 21.6 mm, p=0.005], there were no significant differences in incidences of obstructed defecation (OD) and fecal incontinence (FI) between the groups. Sixteen patients with anterior RAI and 137 patients with circular RAI had OD. There was no significant difference in the CSS scores between the groups. Twelve patients with anterior RAI and 108 patients with circular RAI had FI. No significant difference in the FISI scores between the groups. Approximately one tenth of the whole RAI was anterior in location, and symptoms in patients with anterior RAI were similar to those with circular RAI.


Isolation and characterisation of Vibrio cholerae from fish examined postmortem at ZSL London Zoo between 2014 and 2018.
When suspect Vibrio cholerae were cultured from fish at ZSL London Zoo, investigations were carried out to determine whether they were possible causes of cholera. Bacterial culture was carried out on fish examined postmortem and colonies were identified using standard techniques including the API 20NE biochemical test kits. Suspect isolates were submitted to the Public Health England laboratory for additional testing. Separately, a number of fish were submitted for routine histopathology. On 13 occasions between 2014 and 2018, suspected V cholerae were cultured from individuals of eight different freshwater fish species. Archived cultures for eight of these (from six different fish species) were investigated and seven isolates (from five fish species) were confirmed as V cholerae, but all were non-O1, non-O139 strains. Whole-genome sequencing showed that the five fish species had unique V cholerae multilocus sequence types (three isolates from Aphanius danfordii were identical), all of which were genetically distant from human isolates. There was no evidence that these isolates could cause cholera. Histopathological changes consistent with vibriosis were seen in several fish, suggesting that V cholerae were causing the disease, but there were also concurrent infections or predisposing stress factors.


Approximation algorithms.
Increasing global competition, rapidly changing markets, and greater consumer awareness have altered the way in which corporations do business. To become more efficient, many industries have sought to model some operational aspects by gigantic optimization problems. It is not atypical to encounter models that capture 10(6) separate "yes" or "no" decisions to be made. Although one could, in principle, try all 2(10(6)) possible solutions to find the optimal one, such a method would be impractically slow. Unfortunately, for most of these models, no algorithms are known that find optimal solutions with reasonable computation times. Typically, industry must rely on solutions of unguaranteed quality that are constructed in an ad hoc manner. Fortunately, for some of these models there are good approximation algorithms: algorithms that produce solutions quickly that are provably close to optimal. Over the past 6 years, there has been a sequence of major breakthroughs in our understanding of the design of approximation algorithms and of limits to obtaining such performance guarantees; this area has been one of the most flourishing areas of discrete mathematics and theoretical computer science.


Development of a reverse genetics system for Sosuga virus allows rapid screening of antiviral compounds.
Sosuga virus (SOSV) is a recently discovered zoonotic paramyxovirus isolated from a single human case in 2012; it has been ecologically and epidemiologically associated with transmission by the Egyptian rousette bat (Rousettus aegyptiacus). Bats have long been recognized as sources of novel zoonotic pathogens, including highly lethal paramyxoviruses like Nipah virus (NiV) and Hendra virus (HeV). The ability of SOSV to cause severe human disease supports the need for studies on SOSV pathogenesis to better understand the potential impact of this virus and to identify effective treatments. Here we describe a reverse genetics system for SOSV comprising a minigenome-based assay and a replication-competent infectious recombinant reporter SOSV that expresses the fluorescent protein ZsGreen1 in infected cells. First, we used the minigenome assay to rapidly screen for compounds inhibiting SOSV replication at biosafety level 2 (BSL-2). The antiviral activity of candidate compounds was then tested against authentic viral replication using the reporter SOSV at BSL-3. We identified several compounds with anti-SOSV activity, several of which also inhibit NiV and HeV. Alongside its utility in screening for potential SOSV therapeutics, the reverse genetics system described here is a powerful tool for analyzing mechanisms of SOSV pathogenesis, which will facilitate our understanding of how to combat the potential public health threats posed by emerging bat-borne paramyxoviruses.


Alzheimer's, atherosclerosis, and aggregates: a role for bacterial degradation.
Several of the most prevalent and severe age-related diseases, notably Alzheimer's disease and atherosclerosis, feature the accumulation of non-degradable aggregates within the lysosomes of disease-affected cells. At an early point in disease progression, the breakdown of lysosomal contents by the resident catabolic enzymes stops working properly. A return of lysosomal enzymatic activity to pre-disease levels may restore aggregate elimination. In this review, a method of bioremediation-derived lysosomal enzyme enhancement is proposed, featuring the cellular introduction of microbial-isolated enzymes, or xenoenzymes. The benefits and challenges of using xenoenzymes to break down aggregates are discussed. As the size of our elderly population grows, the incidence of age-related diseases will increase, necessitating the exploration of radical, but potentially powerful, therapeutic strategies.


Precision of vancomycin and daptomycin MICs for methicillin-resistant Staphylococcus aureus and effect of subculture and storage.
The reproducibility of vancomycin and daptomycin MICs, measured by broth microdilution (BMD) and Etest, was prospectively assessed for 10 methicillin-resistant Staphylococcus aureus (MRSA) isolates from the blood samples from patients on vancomycin therapy. The isolates were tested at the time of isolation from blood and following 5, 10, and 20 subcultures and at 1, 3, 6, and 12 months of storage at -70 °C. The MICs were determined by Etest and BMD using two different manufacturers (BBL and Difco) of cation-adjusted Mueller-Hinton broth (CA-MHB), and using three different drug powders: vancomycin from Sigma, vancomycin from Novation, and daptomycin from Cubist. The antimicrobial concentrations tested were 0.25, 0.5, 0.75, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, and 4.0 μg/ml. Two isolates were vancomycin intermediate and daptomycin nonsusceptible, and two isolates had reduced susceptibility to vancomycin (BMD MIC, 1.5 or 2.0 μg/ml). The vancomycin MICs were significantly higher in the BBL CA-MHB than those in the Difco CA-MHB, and with Sigma versus Novation vancomycin powder. The daptomycin MICs were also significantly higher in the BBL CA-MHB. The Etest MICs were significantly higher than those obtained by BMD for vancomycin but not for daptomycin. The average precision of the vancomycin BMD MICs when analyzing 20 results was ± 1.10-fold log2 dilutions, and it was ± 1.67-fold for daptomycin (10 results). The average precision for Etest was ± 1.11-fold for vancomycin and ± 1.16-fold for daptomycin. No significant change in MICs was noted following 5, 10, or 20 subcultures or at up to 6 months of frozen storage. However, the vancomycin MICs alone were significantly lower (0.74-fold) following 12 months of frozen storage. From these data, despite variations in CA-MHB and antimicrobial powder, the MIC result precision was <0.5 log2 dilutions in a single laboratory, suggesting that testing interdilution MICs (e.g., MICs between serial 2-fold dilutions) is a possibility. A more accurate method for measuring vancomycin MIC results is thus possible, but further standardization of BMD testing would be required to achieve this goal.


Resonance scattering particles as biological nanosensors in vitro and in vivo.
Recently dark-field microscopy and Rayleigh scattering spectroscopy have emerged as complementary technologies for ultrasensitive biological detection and imaging with high spatial and temporal resolution. Plasmonic resonant nanoparticles are key nano-scale probes for these technologies that have enabled single-molecule sensitivity and imaging. In this tutorial review, we focus on the use of plasmonic probes as single-particle biological nanosensors in vitro and in vivo. The progress in this field over the last decade will be highlighted.


Applying Neural Manifold Constraint on Point Process Model for Neural Spike Prediction.
Neural prostheses can compensate for functional losses caused by blocked neural pathways by modeling neural activities among cortical areas. Existing methods generally utilize point process models to predict neural spikes from one area to another, and optimize the model by maximizing the log-likelihood between model predictions and recorded activities of individual neurons. However, single-neuron recordings can be distorted, while neuron population activity tends to reside within a stable subspace called the neural manifold, which reflects the connectivity and correlation among output neurons. This paper proposes a neural manifold constraint to modify the loss function for model training. The constraint term minimizes the distance from model predictions to the empirical manifold to amend the model predictions from distorted recordings. We test our methods on synthetic data with distortion on output spike trains and evaluate the similarity between model predictions and original output spike trains by the Kolmogorov-Smirnov test. The results show that the models trained with constraint have higher goodness-of-fit than those trained without constraint, which indicates the potential better approach for neural prostheses in noisy environments.


Circular current in a one-dimensional open quantum ring in the presence of magnetic field and spin-orbit interaction.
In an open quantum system having a channel in the form of loop geometry, the current inside the channel, namely circular current, and overall junction current, namely transport current, can be different. A quantum ring has doubly degenerate eigen energies due to periodic boundary condition that is broken in an asymmetric ring where the ring is asymmetrically connected to the external electrodes. Kramers' degeneracy and spin degeneracy can be lifted by considering non-zero magnetic field and spin-orbit interaction (SOI), respectively. Here, we find that symmetry breaking impacts the circular current conductance vs energy (E) spectra in addition to lifting the degeneracy. For charge and spin current conductances, the corresponding effects are not the same. Under symmetry-breaking they may remain symmetric or anti-symmetric or asymmetric aroundE = 0 whereas the transmission function (which is proportional to the junction current conductance) vs energy characteristic remains symmetric aroundE = 0. This study leads us to estimate the qualitative nature of the circular current and the choices of Fermi-energy/chemical potential to have a net non-zero current. As a result, we may manipulate the system to generate pure currents of charge, spin, or both, which is necessary for any spintronic and electronic applications.


An energetic stellar outburst accompanied by circumstellar light echoes.
Some classes of stars, including novae and supernovae, undergo explosive outbursts that eject stellar material into space. In 2002, the previously unknown variable star V838 Monocerotis brightened suddenly by a factor of approximately 10(4). Unlike a supernova or nova, it did not explosively eject its outer layers; rather, it simply expanded to become a cool supergiant with a moderate-velocity stellar wind. Superluminal light echoes were discovered as light from the outburst propagated into the surrounding, pre-existing circumstellar dust. Here we report high-resolution imaging and polarimetry of those light echoes, which allow us to set direct geometric distance limits to the object. At a distance of >6 kpc, V838 Mon at its maximum brightness was temporarily the brightest star in the Milky Way. The presence of the circumstellar dust implies that previous eruptions have occurred, and spectra show it to be a binary system. When combined with the high luminosity and unusual outburst behaviour, these characteristics indicate that V838 Mon represents a hitherto unknown type of stellar outburst, for which we have no completely satisfactory physical explanation.