Dose-response effects of TPI ASM8 in asthmatics after allergen.
TPI ASM8 contains two modified antisense oligonucleotides (AON) targeting the beta subunit (β(c) ) of the IL-3, IL-5, GM-CSF receptors and the chemokine receptor CCR3. A previous study suggested that TPI ASM8 had broader effects than just inhibition of eosinophils in asthmatics. We assessed whether TPI ASM8 caused a dose-dependent attenuation in the inflammatory and physiological changes after inhaled allergen challenge (AIC). This single-center, open-label, stepwise-ascending dose study was conducted in fourteen stable, mild allergic asthmatics. Following placebo AIC, subjects underwent AIC after 4 days treatment with 1, 2, and 4 mg BID and finally 8 mg once daily (OD) of TPI ASM8, inhaled via the I-Neb™ nebuliser. Treatments were separated by 2-3-week washout periods. TPI ASM8 was safe and well tolerated at all doses. TPI ASM8 8 mg OD reduced eosinophils in sputum after AIC (by 60.9% at 7 h and 68.4% at 24 h post-AIC, P=0.016 and P=0.007, respectively). Additionally, TPI ASM8 8 mg OD significantly attenuated the early and late airway responses as shown by the reduction in the area under the curve by 45% (P=0.016) and 59%, (P=0.0015), respectively, the increase in eosinophil cationic protein (ECP) by up to 57% (P=0.021), and airway responsiveness to methacholine by more than 1 doubling dose (P=0.012). A dose-response relationship was noted, and efficacy was maintained with once per day administration. TPI ASM8 attenuated a broad range of inflammatory and physiological changes after AIC, suggesting that CCR3, IL-3, and GM-CSF also are important targets for the management of asthma.


Application of reminiscence treatment on older people with dementia: a case study in Pingtung, Taiwan.
Reminiscence therapy has been utilized for many years in the treatment of dementia in older people. Purposes of the research included examining different methods of promoting interactivity, social participation, cognitive function improvement in those with dementia, and the effectiveness in reducing symptoms of depression following group treatment. This study used pretest and posttest electroencephalography (EEG) measurements to test reminiscence therapy efficacy on participants. This research organized a social group work with 12 elderly clients with dementia (mild to moderate stage) selected from among 90 residents of an older persons care facility in Pingtung. Eleven agreed to join the study, and 10 completed successfully all treatment sessions. Eight sessions of reminiscence cooking lessons were conducted. The effectiveness of interventions was evaluated by comparing presession and postsession EEG, mental health status, depression scale, and feeling of participation scale scores. Significant differences in values, particularly for EEG, were found between the two sets of scores. The average value of participants' fast waves rose from 43.88 to 55.12, whereas average slow-wave values fell from 56.12 to 44.13. After analysis using the Wilcoxon matched paired signed rank test, significant differences were noted. Findings and suggestions include the following: (a) The rise in Mini-Mental State Examination and reduction in depression scale scores, although noted, were not significant, and (b) the self-achievement, emotional stability, family atmosphere, and physical needs of participants were met. The authors recommend that reminiscence group work be promoted in the home for older persons and that childhood cooking sessions twice each week may be the ideal format for reminiscence group work.


Plasma 25-hydroxyvitamin D concentrations in preterm infants receiving oral vitamin D supplements.
Plasma 25-hydroxy-cholecalciferol (25-OHD) was measured serially in two groups of preterm infants receiving either 400 IU or 1000 IU oral vitamin D3 daily. All the babies were able to absorb and hydroxylate vitamin D3 adequately by 36 weeks' gestational age. The higher daily supplement had no advantage over the lower dose.


Femoral nerve palsy secondary to traumatic iliacus haematoma--a young athlete's injury.
Femoral neuropathy secondary to iliacus haematoma in the absence of a bleeding diathesis is rare. Of the few documented cases in the literature, most have occurred in adolescents participating in sport. We report a case of femoral nerve palsy complicating an iliacus haematoma occurring in a young person with minimal trauma that resolved with non-operative management. Although rare, iliacus haematoma should be included in the differential diagnosis of groin pain as the number of children and young adults participating in sport increases.


The role of occupational therapy in the Solomon Islands: experiences and perceptions of occupational therapists and rehabilitation health workers.
In most Western countries occupational therapy is well established as a crucial aspect of overall health care; however, in low-middle-income countries it is still an emerging profession. This article investigates the role of occupational therapy in the Solomon Islands by examining the experiences and perceptions of occupational therapists (OTs) and other rehabilitation health workers who have worked there. Using a qualitative research design, participants were recruited using purposive sampling, and data were gathered through in-depth interviews, diary entries and observations of an occupational therapy setting in the Solomon Islands. Ten participants were interviewed. A significant role in health-promoting practices was revealed through the various tasks undertaken by OTs in the Solomon Islands, including influencing health policy and practice broadly through advocacy and education. Challenges relating to geography, cultural, language and resource barriers, and professional practice issues were identified. Implications for practice were drawn from the data including the need for culturally safe practice, pragmatism, creativity and practising across disciplinary boundaries. Results illustrate a clear role for occupational therapy in promoting health and sustainability of rehabilitation therapy services in the Solomon Islands, which also have relevance throughout Pacific island nations.


Matching catalytic activity to developmental function: tolloid-related processes Sog in order to help specify the posterior crossvein in the Drosophila wing.
The Drosophila tolloid (tld) and tolloid related (tlr) gene products belong to a family of developmentally important proteases that includes Bone Morphogenetic Protein 1 (Bmp1). Tld is required early in Drosophila development for proper patterning of dorsal embryonic structures, whereas Tlr is required later during larval and pupal stages of development. The major function of Tld is to augment the activity of Decapentaplegic (Dpp) and Screw (Scw), two members of the Bmp subgroup of the Tgf beta superfamily, by cleaving the Bmp inhibitor Short gastrulation (Sog). In this study, we provide evidence that Tlr also contributes to Sog processing. Tlr cleaves Sog in vitro in a Bmp-dependent manner at the same three major sites as does Tld. However, Tlr shows different site selection preferences and cleaves Sog with slower kinetics. To test whether these differences are important in vivo, we investigated the role of Tlr and Tld during development of the posterior crossvein (PCV) in the pupal wing. We show that tlr mutants lack the PCV as a result of too little Bmp signaling. This is probably caused by excess Sog activity, as the phenotype can be suppressed by lowering Sog levels. However, Tld cannot substitute for Tlr in the PCV; in fact, misexpressed Tld can cause loss of the PCV. Reducing levels of Sog can also cause loss of the PCV, indicating that Sog has not only an inhibitory but also a positive effect on signaling in the PCV. We propose that the specific catalytic properties of Tlr and Tld have evolved to achieve the proper balance between the inhibitory and positive activities of Sog in the PCV and early embryo, respectively. We further suggest that, as in the embryo, the positive effect of Sog upon Bmp signaling probably stems from its role in a ligand transport process.


Effectiveness of miltefosine in cutaneous leishmaniasis caused by Leishmania tropica in Pakistan after antimonial treatment failure or contraindications to first line therapy-A retrospective analysis.
Cutaneous leishmaniasis (CL) is a neglected tropical skin disease, caused by Leishmania protozoa. In Pakistan, where CL caused by L. tropica is highly endemic, therapy with pentavalent antimonials is the standard of care, but has significant toxicity when used in systemic therapy, while are no evidence-based safer alternative treatment options for L. tropica. The efficacy of oral miltefosine has not been studied in CL caused by L. tropica. We evaluated effectiveness and tolerability of miltefosine in patients with previous treatment failure or with contraindications to systemic antimonial treatment. A retrospective review was conducted of a cohort of CL patients who were treated with a 28-day course of miltefosine between December 2017 and August 2019, in urban Quetta, Pakistan, an area endemic for L. tropica. Descriptive analyses were performed, and effectiveness was assessed by initial response after treatment, and final cure at routine follow up visits, six weeks to three months post-treatment. Tolerability was assessed by routinely reported adverse events. Of the 76 CL patients in the cohort, 42 (55%) had contraindications to systemic antimonial treatment, and 34 (45%) had failure or relapse after antimonial treatment. Twelve patients defaulted during treatment and 12 patients were lost to follow up. In the remaining 52 patients, final cure rate was 77% (40/52). In those with contraindications to systemic antimonial treatment the final cure rate was 83% (24/29) and in the failure and relapse group 70% (16/23). Twenty-eight patients (40.0%) reported 39 mild to moderate adverse events with the main complaints being nausea (41.0%), general malaise (25.6%), and stomach pain (12.8%). Results indicate that miltefosine is an effective second line treatment in CL in areas endemic for L. tropica. Prospective studies with systematic follow up are needed to obtain definitive evidence of effectiveness and tolerability, including identification of risk factors for miltefosine treatment failure.


Terlipressin therapy for renal failure in cirrhosis.
Renal failure is common in cirrhosis frequently due to hepatorenal syndrome (HRS). Terlipressin and albumin improve renal function with a trend to prolong survival in HRS, but prognostic factors with therapy have been poorly studied. Forty-five cirrhotics seen consecutively in a single centre with renal failure defined as oliguria/anuria and/or rising creatinine and no response to volume loading, without intrinsic renal disease, sepsis, gastrointestinal bleeding [median Child-Pugh score 12(8-14)/Model for End-Stage Liver Disease 29(10-40)], had intravenous terlipressin and albumin and were audited retrospectively classified into three groups: group 1 HRS type 1 (15), group 2 HRS type 2 (11) and group 3(19): not fulfilling HRS 1 or 2 criteria. Baseline median creatinine was 1.7 (0.9-5.46) mg/dl and 30 (67%) had creatinine greater than 1.5 mg/dl. All 45 patients had initial colloid/albumin and 31 continued terlipressin (2-4 mg/day) for a median 8 (2-76) days. Improvement in serum creatinine occurred in 23 (51%) [(1.3 mg/dl (0.6-3.9)] compared with baseline [1.7 mg/dl (0.92-3.75)] (P<0.001). In the multivariate analysis a greater reduction in creatinine between baseline and day 4 (95% confidence interval, odds ratio: 0.25) was associated with improved survival at 6 weeks. Albumin and terlipressin improve renal failure in the absence of sepsis in cirrhosis independently of whether HRS criteria are fulfilled or not. Improvement at 4 days of therapy is associated with better survival. Randomized studies are needed for oliguria and rising creatinine in cirrhotics even if HRS criteria are not fulfilled.


Identification of genes with differential regulation in primate endometrium during the proliferative and secretory phases of the cycle.
To study gene expression in the endometrium at different stages of the menstrual cycle, differential mRNA display and reverse Northern analysis were performed on uterine tissues from cynomolgus monkeys. Eutopic endometrial RNA was prepared from uteri of animals that were either ovariectomized and supplemented with hormones, or were not ovariectomized but were subjected to surgically induced endometriosis. A number of genes were identified whose levels fluctuated between the proliferative and secretory phases of the cycle. Expression of four genes thus identified was further examined by in situ hybridization to normal human endometrial biopsies. Iodothyronine deiodinase was uniformly expressed at all stages of the human cycle that were studied: proliferative, secretory, and menstrual. Fibulin 1, osteopontin, and cathepsin H exhibited complex temporal and spatial regulation. Fibulin 1 was expressed in glandular epithelia during the menstrual phase, but expression switched to the stroma during the secretory phase. During the menstrual phase, osteopontin was expressed at high levels in glandular epithelia and in isolated stromal cells that may be of immune origin. Secretory phase expression of osteopontin was confined to a sub-population of epithelial cells. Cathepsin H was expressed in proliferative and menstrual phase endometrium, but expression disappeared in the secretory phase. Messenger RNA for fibulin 1, osteopontin, and iodothyronine deiodinase was detected in an endometriosis sample. Our data support functional roles for fibulin 1, osteopontin, cathepsin H and thyroid hormone in endometrium.


Mapping of Stomach, Colorectal, and Bladder Cancers in Iran, 2004-2009: Applying Bayesian Polytomous Logit Model.
According to the last report of Iran cancer registry, stomach, colorectal, and bladder cancers are the most prevalent cancers. The present study focused on separating the latent risk surface into shared and disease-specific components. In this study, data consisting of stomach, colorectal, and bladder cancers in 30 provinces of Iran during 2004-2009 are considered. These data are analyzed by polytomous logit model. The incidence of stomach cancer acts as the reference category (the surrogate for smoking). Then, the log odds are decomposed into shared and specific structured spatial and unstructured spatial components. These latent components help to detect spatial patterns of shared and disease-specific risk factors. Central, Southern, Eastern, and Southwestern provinces are supposed as high-risk regions for shared risk factor for colorectal and bladder cancers. This shared risk factor is slightly associated more with bladder than with colorectal cancer. Northern, northwestern, and central regions and also three borderline provinces in southwestern are high-risk regions for colorectal cancer. Central, eastern, southern, and western strip of the country except Ilam are found as the high-risk regions of bladder cancer. After considering known shared risk factor of the three cancers, it turns out that colorectal and bladder cancers have unknown shared risk factor. The significant difference in their lifestyle and eating habits could be an assumption of the risk factor.


Embedded human control of robots using myoelectric interfaces.
Myoelectric controlled interfaces have become a research interest for use in advanced prostheses, exoskeletons, and robot teleoperation. Current research focuses on improving a user's initial performance, either by training a decoding function for a specific user or implementing "intuitive" mapping functions as decoders. However, both approaches are limiting, with the former being subject specific, and the latter task specific. This paper proposes a paradigm shift on myoelectric interfaces by embedding the human as controller of the system to be operated. Using abstract mapping functions between myoelectric activity and control actions for a task, this study shows that human subjects are able to control an artificial system with increasing efficiency by just learning how to control it. The method efficacy is tested by using two different control tasks and four different abstract mappings relating upper limb muscle activity to control actions for those tasks. The results show that all subjects were able to learn the mappings and improve their performance over time. More interestingly, a chronological evaluation across trials reveals that the learning curves transfer across subsequent trials having the same mapping, independent of the tasks to be executed. This implies that new muscle synergies are developed and refined relative to the mapping used by the control task, suggesting that maximal performance may be achieved by learning a constant, arbitrary mapping function rather than dynamic subject- or task-specific functions. Moreover, the results indicate that the method may extend to the neural control of any device or robot, without limitations for anthropomorphism or human-related counterparts.


Flow cytometric monitoring of bacterioplankton phenotypic diversity predicts high population-specific feeding rates by invasive dreissenid mussels.
Species invasion is an important disturbance to ecosystems worldwide, yet knowledge about the impacts of invasive species on bacterial communities remains sparse. Using a novel approach, we simultaneously detected phenotypic and derived taxonomic change in a natural bacterioplankton community when subjected to feeding pressure by quagga mussels, a widespread aquatic invasive species. We detected a significant decrease in diversity within 1 h of feeding and a total diversity loss of 11.6 ± 4.1% after 3 h. This loss of microbial diversity was caused by the selective removal of high nucleic acid populations (29 ± 5% after 3 h). We were able to track the community diversity at high temporal resolution by calculating phenotypic diversity estimates from flow cytometry (FCM) data of minute amounts of sample. Through parallel FCM and 16S rRNA gene amplicon sequencing analysis of environments spanning a broad diversity range, we showed that the two approaches resulted in highly correlated diversity measures and captured the same seasonal and lake-specific patterns in community composition. Based on our results, we predict that selective feeding by invasive dreissenid mussels directly impacts the microbial component of the carbon cycle, as it may drive bacterioplankton communities toward less diverse and potentially less productive states.


Mechanism of long-lasting synaptic inhibition in Aplysia neuron R15.
1. Long-lasting inhibition is a synaptically mediated response found in certain molluscan nerve cells that fire action potentials in bursts. It is elicited by repetitive stimulation of a presynaptic nerve and may last for minutes or hours after stimulation. 2. Voltage-clamp techniques were employed to measure the voltage dependence of the synaptically elicited current. Current-voltage curves were obtained by stepping or sweeping the voltage over the range -40 to -120 mV. 3. Long-lasting inhibition was found to be mediated by two separate conductance mechanisms. A component that reverses near -80 mV is most prominent at times up to 5 min following stimulation. A component with no reversal potential between -40 and -120 mV predominates at later times. 4. The reversible component is attenuated by reducing the intensity of stimulation of the presynaptic nerve, by injection of TEA into the postsynaptic cell, or by activation of a potassium conductance with serotonin prior to stimulation of the nerve. Thus, the reversible component appears to be mediated by an increase in potassium conductance. 5. The effects of the nonreversible component measured in the soma appear to be too large to attribute it to a conductance change that is electrically "distant" from the soma. It is attenuated by turning off a resting inward ion conductance with dopamine prior to stimulation of the nerve. It is not affected by short exposure to ouabain, but is attenuated by longer exposures that reduce the sodium and calcium gradients. Thus, the nonreversible component may be mediated by a decrease in voltage-dependent inward current flow carried by sodium or calcium.


The presumptive magnetosome protein Mms16 is a poly(3-hydroxybutyrate) granule-bound protein (phasin) in Magnetospirillum gryphiswaldense.
The Mms16 protein has been previously found to be associated with isolated magnetosomes from two Magnetospirillum strains. A function of this protein as a magnetosome-specific GTPase involved in the formation of intracellular magnetosome membrane vesicles was suggested. Here we present a study of the Mms16 protein from Magnetospirillum gryphiswaldense to clarify its function. Insertion-duplication mutagenesis of the mms16 gene did not affect the formation of magnetosome particles but resulted in the loss of the ability of M. gryphiswaldense cell extracts to activate poly(3-hydroxybutyrate) (PHB) depolymerization in vitro, which was coincident with loss of the most abundant 16-kDa polypeptide from preparations of PHB granule-bound proteins. The mms16 mutation could be functionally complemented by enhanced yellow fluorescent protein (EYFP) fused to ApdA, which is a PHB granule-bound protein (phasin) in Rhodospirillum rubrum sharing 55% identity to Mms16. Fusions of Mms16 and ApdA to enhanced green fluorescent protein (EGFP) or EYFP were colocalized in vivo with the PHB granules but not with the magnetosome particles after conjugative transfer to M. gryphiswaldense. Although the Mms16-EGFP fusion protein became detectable by Western analysis in all cell fractions upon cell disruption, it was predominantly associated with isolated PHB granules. Contrary to previous suggestions, our results argue against an essential role of Mms16 in magnetosome formation, and the previously observed magnetosome localization is likely an artifact due to unspecific adsorption during preparation. Instead, we conclude that Mms16 in vivo is a PHB granule-bound protein (phasin) and acts in vitro as an activator of PHB hydrolysis by R. rubrum PHB depolymerase PhaZ1. Accordingly, we suggest renaming the Mms16 protein of Magnetospirillum species to ApdA, as in R. rubrum.


The molecular dimension of microbial species: 2. Synechococcus strains representative of putative ecotypes inhabiting different depths in the Mushroom Spring microbial mat exhibit different adaptive and acclimative responses to light.
Closely related strains of thermophilic Synechococcus were cultivated from the microbial mats found in the effluent channels of Mushroom Spring, Yellowstone National Park (YNP). These strains have identical or nearly identical 16S rRNA sequences but are representative of separate, predicted putative ecotype (PE) populations, which were identified by using the more highly resolving psaA locus and which predominate at different vertical positions within the 1-mm-thick upper-green layer of the mat. Pyrosequencing confirmed that each strain contained a single, predominant psaA genotype. Strains differed in growth rate as a function of irradiance. A strain with a psaA genotype corresponding to a predicted PE that predominates near the mat surface grew fastest at high irradiances, whereas strains with psaA genotypes representative of predominant subsurface populations grew faster at low irradiance and exhibited greater sensitivity to abrupt shifts to high light. The high-light-adapted and low-light-adapted strains also exhibited differences in pigment content and the composition of the photosynthetic apparatus (photosystem ratio) when grown under different light intensities. Cells representative of the different strains had similar morphologies under low-light conditions, but under high-light conditions, cells of low-light-adapted strains became elongated and formed short chains of cells. Collectively, the results presented here are consistent with the hypothesis that closely related, but distinct, ecological species of Synechococcus occupy different light niches in the Mushroom Spring microbial mat and acclimate differently to changing light environments.


Traumatic cataract in patient with anterior megalophthalmos: Case report.
Megalophthalmos anterior is a rare, bilateral, nonprogressive, hereditary, congenital disorder, characterized by the enlargement of all anterior segment structures of the eye, with megalocornea, iris atrophy, and zonular abnormalities commonly found. Usually almost asymptomatic in young patients, with most complaints concerning blurred vision due to the common corneal astigmatism, it might in time lead to several complications including premature cataract formation and pigmentary glaucoma. This review presents the case of a 47-year old patient referred to our clinic for traumatic cataract surgery, with striking bilateral megalocornea, somehow overlooked during previous ophthalmic examinations in his local outpatient clinic. We noticed markedly enlarged corneas and deepened anterior chambers of his both eyes, accompanied by intumescent, white cataract of the right eye, and incipient cortical cataract of the left eye. Best corrected visual acuity (BCVA) was counting fingers in the right eye and 20/25 in the left eye. Additional examination revealed multiple abnormalities of the anterior segment, leading to the diagnosis of anterior megalophthalmos. It is astounding the patient remained undiagnosed during previous examinations, with his megalocornea and remarkably deep anterior chamber so apparent. We performed standard phacoemulsification procedure, with 3 piece posterior chamber intraocular lens (PCIOL) implantation into the lens capsule. The surgery was uneventful, with postoperative BCVA of 20/20 in the right eye, and no dislocation of the lens in 9-month observation period. Complicated cataract in patients with anterior megalophthalmos can be successfully treated with standard phacoemulsification procedure followed by the bag PCIOL implantation.Care needs to be taken during cataract surgery in these patients, as zonular abnormalities and lens enlargement are common, resulting in increased rate of intra- and postoperative complications. As patients with anterior megalophthalmos require a more careful follow-up.


Reduction of intake in the rat due to gastric filling.
With two exceptions, the literature shows that confining ingested fluid to the stomach of a rat during an intake test had no effect on the volume ingested. In the two exceptions (J. D. Davis and J. L. Sayler, Physiol. Behav. In press. J. A. Deutsch, Handbook of Behavioral Neurobiology. Neurobiology of Food and Fluid Intake, edited by E. M. Stricker. New York: Plenum, 1990, vol. 10), intake with the cuff open was very large, suggesting that, when intake on cuff-open tests exceeds some critical volume, confining all of it to the stomach during the test will reduce intake. To test this, we measured intake of three different solutions known to stimulate large intake with the pylorus open and closed. In cuff-closed tests, intake was less than in cuff-open tests. In cuff-closed tests, rate of licking began to decline within 5-6 min when only about one-quarter of the ultimate contents of the stomach had accumulated, indicating that some signal from the stomach slowed the rate of ingestion before the full capacity of the stomach was reached. This shows that the stomach is sensitive to its contents when it contains only a small proportion of its capacity.


Evaluation of a 5'-nuclease (TaqMan) assay for the detection of virulent strains of Yersinia enterocolitica in raw meat and tofu samples.
Culture methods for detecting virulent Yersinia enterocolitica require selective enrichment and a series of confirmatory tests that are time-consuming, costly, and laborious. The objective of this study was to evaluate a fluorogenic 5'-nuclease assay for detecting the enterotoxin yst gene of virulent Y. enterocolitica in pure cultures, inoculated ground pork samples, and naturally contaminated food samples. These results were then compared with "gold standard" methods recommended by the U.S. Food and Drug Administration in the Bacteriological Analytical Manual for detecting pathogenic Y. enterocolitica. The 5'-nuclease assay was able to identify the organism in 100% of the repetitions when 10(2) CFU/ml or more organisms were present in pure cultures and 10(3) CFU/g or more organisms were present in ground pork. Similar recovery efficiency on cefsulodin-irgasan-novobiocin (CIN) agar plates was only evident when 10(5) CFU/ml or more organisms were present in pure culture and 10(6) CFU/g or more organisms were present in inoculated ground pork. The 5'-nuclease assay indicated a contamination rate of 35.5% (94/265) in various meats and tofu, whereas the CIN plating method indicated a contamination rate of 28.3% (75/265). This resulted in 100% sensitivity and 64.5% specificity for the 5'-nuclease assay when compared with the standard culture recovery method. Only 75% (60/80) of the Yersinia spp. isolated on CIN was identified as containing a virulence plasmid by autoagglutination and crystal violet binding tests. These results indicate that the true rate of contamination of virulent Y. enterocolitica in pork and other processed meats and foods is being underestimated using current detection methods. This study demonstrates the potential of the 5'-nuclease assay for rapidly and specifically detecting virulent Y. enterocolitica in processed foods with the added advantage of being an automated detection system with high-throughput capability.


[Biochemical blood serum indexes in rats with hypothyroidism corrected by iodide-100 preparation].
In experiments performed on rats with hypothyrosis, the blood serum indexes (the level of triodthyronine, thyroxin, thyrothropic hormone of adenohypophysis, the level of cholesterol, triglycerides, nitric oxide) and the level of iodine excretion with urine before and after correction of iodine deficiency by "Iodide-100" drug has been studied. It has been revealed that hypothyrosis has negative influence on lipid metabolism indexes and is accompanied by decrease in nitric oxide level in blood serum. This could lead to destabilization of structure and functions of cells of different tissues of the organism. Administration of the drug "Iodide-100", leads to stabilization of hormonal and lipid states and nitric oxide level in blood serum increases.


Phosphorylation of mitogen-activated protein kinase cascade during early embryo development in the mouse.
The mitogen-activated protein kinase (MAPK) cascade is one of the most important signal transduction pathways that regulate the cell cycle in somatic cells. The present study examined the phosphorylation states of components in the MAPK cascade, Raf-1, MEK-1, and extracellular signal regulated kinases (ERKs), which are activated by mitogens, throughout early mouse embryo development and in cultured somatic cells generally. In somatic cells, Raf-1 and MEK-1 were phosphorylated at M-phase and dephosphorylated during interphase. ERKs were not phosphorylated at any stage during the cell cycle. These results were similar to previous findings for the first and second cell cycles of early mouse embryos. In contrast, after the four-cell stage, not only ERKs, but also Raf-1 and MEK-1, were not phosphorylated at any stage during the cell cycle in mouse early embryos. These results suggest that the MAPK cascade in mouse embryos is regulated by the same mechanism as in somatic cells before the two-cell stage, and that regulation is changed to an embryo-specific mechanism after the four-cell stage.


A molecular link between FGF and Dpp signaling in branch-specific migration of the Drosophila trachea.
The tracheal system of Drosophila embryos achieves its archetypal branching pattern through a series of cell migration events requiring the FGF, Dpp, and Wg/WNT signaling pathways. To gain insight into tracheal cell migration, we performed an F4 EMS mutagenesis screen to generate and characterize new mutations resulting in tracheal defects. From 2591 mutagenized third chromosome lines, we identified 33 mutations with defects in tracheal development, corresponding to 12 distinct complementation groups. The new mutations included novel hypomorphic alleles of the FGF receptor gene, breathless, and the ETS-domain transcription factor gene, pointed. We show that reduced function of either breathless or pointed specifically affects migration of the dorsal and ventral tracheal branches, more specific functions than previously described for these genes. Our analysis reveals that breathless and pointed control dorsal branch migration through transcriptional regulation of the Dpp pathway effectors, Knirps and Knirps-related, which are necessary for migration of this branch. We further show that expression of knirps or knirps-related rescues dorsal but not ventral branch migration in the breathless hypomorph. These studies support a model in which both the Dpp- and the FGF-signaling pathways control expression of knirps and knirps-related, thereby regulating cell migration during dorsal branch formation.


Effect of IL-10 and TNF-α on the competence and cryosurvival of in vitro produced Bos indicus embryos.
In vitro-produced embryos are constantly exposed to stressful conditions that can lead to the activation of the apoptotic pathway. The nuclear Kappa B factor (NF-κB) is an inflammatory mediator that induces the expression of tumor necrosis factor (TNF-α), a pro-inflammatory cytokine, while interleukin-10 (IL-10), an anti-inflammatory cytokine, inhibits NF-κB activity. This study aimed to investigate the effects of IL-10 and TNF-α on the competence and cryosurvival of in vitro-produced bovine embryos. Embryos were produced in vitro using standard protocols, and Grade I blastocysts were vitrified using the Cryotop method. Non-vitrified and vitrified blastocysts were subjected to the TUNEL assay. In Experiment I, on day 6.5 (156 h post-insemination), the embryos were treated with PBS (control), 50 ng/mL of IL-10, or a combination of 25 ng/mL of TNF-α and 50 ng/mL of IL-10. Embryonic development and apoptotic rates were monitored. In Experiment II, the same groups were set up, with the addition of a group treated with 25 ng/mL of TNF-α alone. Grade I blastocysts were vitrified 5 h after treatment, and cryosurvival was monitored at until 48 h post-warming. The apoptosis rate and total cell number were investigated in the vitrified-hatched blastocysts. IL-10 alone did not affect developmental competence or cryosurvival (P > 0.05). The IL-10-treated embryos, when exposed in combination with TNF-α, presented a detrimental effect (P < 0.05) in the embryonic development of non-vitrified embryos. However, vitrified blastocysts had no negative effect (P > 0.05). The TNF-α treatment reduced (P < 0.05) the re-expansion rate at 6 h post-warming and increased (P < 0.05) the apoptosis rate in vitrified hatched blastocysts, whereas no effect (P > 0.05) of the treatments was detected in the hatching rate and total cell number post-warming. In conclusion, TNF-α has a detrimental effect on embryonic developmental competence and cryosurvival by compromising the development of non-vitrified embryos and apoptotic-related events of vitrified blastocysts, whereas IL-10, when in combination with TNF-α, appears to attenuate the detrimental effects of TNF-α.


Exercise-based rehabilitation after hospital discharge for survivors of critical illness with intensive care unit-acquired weakness: A pilot feasibility trial.
The aim of this study was to investigate feasibility of exercise-based rehabilitation delivered after hospital discharge in patients with intensive care unit-acquired weakness (ICU-AW). Twenty adult patients, mechanically ventilated for more than 48 hours, with ICU-AW diagnosis at ICU discharge were included in a pilot feasibility randomized controlled trial receiving a 16-session exercise-based rehabilitation program. Twenty-one patients without ICU-AW participated in a nested observational cohort study. Feasibility, clinical, and patient-centered outcomes were measured at hospital discharge and at 3 months. Intervention feasibility was demonstrated by high adherence and patient acceptability, and absence of adverse events, but this must be offset by the low proportion of enrolment for those screened. The study was underpowered to detect effectiveness of the intervention. The use of manual muscle testing for the diagnosis of ICU-AW lacked robustness as an eligibility criterion and lacked discrimination for identifying rehabilitation requirements. Process evaluation of the trial identified methodological factors, categorized by "population," "intervention," "control group," and "outcome." Important data detailing the design, conduct, and implementation of a multicenter randomized controlled trial of exercise-based rehabilitation for survivors of critical illness after hospital discharge have been reported. Clinical Trials Identifier NCT00976807.


α-Glucosidase Inhibitory Activity of Fermented Okara Broth Started with the Strain Bacillus amyloliquefaciens SY07.
In this work, a new strain of Bacillus amyloliquefaciens SY07 isolated from a traditional fermented soybean food was reported to possess remarkable α-glucosidase inhibitor-producing ability. Different culture media were applied for the proliferation of B. amyloliquefaciens SY07, and it was found that fermented okara broth presented the highest α-glucosidase inhibitory activity, while Luria-Bertani medium showed a negative effect. The extract from fermented okara broth acted in a dose-dependent manner to inhibit α-glucosidase activity, with an IC50 value of 0.454 mg/mL, and main inhibitors in the fermentation extract presented a reversible, uncompetitive pattern according to Lineweaver-Burk plots. Moreover, 1-deoxynojirimycin, a recognized α-glucosidase inhibitor, was found in the extract. Results indicated that B. amyloliquefaciens SY07 could utilize okara, a by-product from the soy processing industry, to generate α-glucosidase inhibitors effectively, and be regarded as a novel excellent microbial candidate for safe, economical production of potential functional foods or ingredients with hypoglycemic effect.


Sexually Transmitted Infection Knowledge among Older Adults: Psychometrics and Test-Retest Reliability.
Sexually transmitted infections (STI) among older adults have dramatically increased in recent years, especially among those who are widowed and divorced. The purposes of this study were to: (1) identify STI-related knowledge among older adults; (2) report the psychometric properties of a tool commonly used to assess STI-related knowledge among younger populations using data from adults 65 years and older; and (3) determine test-retest reliability of the tool. Data were analyzed from 43 adults, aged 65-94 years, using the 27-item Sexually Transmitted Disease Knowledge Questionnaire (STD-KQ). Participants completed identical instruments on two separate days with approximately two weeks between. After responses were coded for correctness, composite scores were created. Cronbach's reliability coefficients were calculated to determine response consistency, and Pearson's r coefficients were used to assess test-retest reliability. Of 27 possible correct answers, participants reported an average of 11.47 (±6.88) correct responses on Day 1 and 11.67 (±7.33) correct responses on Day 2. Cronbach's alpha coefficients for the 27-item composite scale were high for both days (0.905 and 0.917, respectively), which indicates strong response consistency. Pearson's r coefficients were high between responses for the 27-item composite scale on Days 1 and 2 (r = 0.882, P < 0.01), which indicates strong test-retest reliability. Pearson's r coefficients were high between responses for all but three of the 27 items when assessed separately. Findings suggest the utility of the STD-KQ to assess STI knowledge among older adults. However, the consistently low knowledge scores highlight the need for educational interventions among this population.


Pericytes contribute to airway remodeling in a mouse model of chronic allergic asthma.
Myofibroblast accumulation, subepithelial fibrosis, and vascular remodeling are complicating features of chronic asthma, but the mechanisms are not clear. Platelet-derived growth factors (PDGFs) regulate the fate and function of various mesenchymal cells and have been implicated as mediators of lung fibrosis. However, it is not known whether PDGF-BB signaling via PDGFRβ, which is critical for the recruitment of pericytes to blood vessels, plays a role in airway remodeling in chronic asthma. In the present study, we used a selective PDGFRβ inhibitor (CP-673451) to investigate the role of PDGFRβ signaling in the development of airway remodeling and lung dysfunction in an established mouse model of house dust mite-induced chronic allergic asthma. Unexpectedly, we found that pharmacological inhibition of PDGFRβ signaling in the context of chronic aeroallergen exposure led to exacerbated lung dysfunction and airway smooth muscle thickening. Further studies revealed that the inflammatory response to aeroallergen challenge in mice was associated with decreased PDGF-BB expression and the loss of pericytes from the airway microvasculature. In parallel, cells positive for pericyte markers accumulated in the subepithelial region of chronically inflamed airways. This process was exacerbated in animals treated with CP-673451. The results indicate that perturbed PDGF-BB/PDGFRβ signaling and pericyte accumulation in the airway wall may contribute to airway remodeling in chronic allergic asthma.


A novel FIP1L1-PDGFRA mutant destabilizing the inactive conformation of the kinase domain in chronic eosinophilic leukemia/hypereosinophilic syndrome.
The Fip1-like-1-platelet-derived growth factor receptor alpha (FIP1L1-PDGFRA) gene fusion is a common cause of chronic eosinophilic leukemia (CEL)/hypereosinophilic syndrome (HES), and patients suffering from this particular subgroup of CEL/HES respond to low-dose imatinib therapy. However, some patients may develop imatinib resistance because of an acquired T674I mutation, which is believed to prevent drug binding through steric hindrance. In an imatinib resistant FIP1L1-PDGFRA positive patient, we analyzed the molecular structure of the fusion gene and analyzed the effect of several kinase inhibitors on FIP1L1-PDGFRA-mediated proliferative responses in vitro. Sequencing of the FIP1L1-PDGFRA fusion gene revealed the occurrence of a S601P mutation, which is located within the nucleotide binding loop. In agreement with the clinical observations, imatinib did not inhibit the proliferation of S601P mutant FIP1L1-PDGFRA-transduced Ba/F3 cells. Moreover, sorafenib, which has been described to inhibit T674I mutant FIP1L1-PDGFRA, failed to block S601P mutant FIP1L1-PDGFRA. Structural modeling revealed that the newly identified S601P mutated form of PDGFRA destabilizes the inactive conformation of the kinase domain that is necessary to bind imatinib as well as sorafenib. We identified a novel mutation in FIP1L1-PDGFRA resulting in both imatinib and sorafenib resistance. The identification of novel drug-resistant FIP1L1-PDGFRA variants may help to develop the next generation of target-directed compounds for CEL/HES and other leukemias.


Hormone-brain-aging relationships, broadly reactive with imidazole-containing dipeptides: targeting of telomere attrition as an aging biomarker and dynamic telomerase activity flirting.
It has been documented that telomere-associated cellular senescence may contribute to certain age-related disorders, and telomere length (TL) may be an informative biomarker of healthy aging. Hormone-brain-aging behavior-modulated telomere dynamics and changes in telomerase activity are consistent elements of cellular alterations associated with changes in proliferative state, and these processes are consequently considered as the new therapeutic drug targets for physiological control with advanced drug delivery and nutritional formulations. We raise and support a therapeutic concept of using nonhydrolyzed forms of naturally occurring neuron-specific imidazole dipeptide-based compounds carnosine and carcinine, making it clinically possible that slowing down the rate of telomere shortening could slow down the human aging process in specific tissues where proliferative senescence is known to occur, with the demonstrated evidence of telomere shortening that appeared to be a hallmark of oxidative stress and disease. Carnosine released from skeletal muscle during exercise may be transported into the hypothalamic tuberomammillary nucleus (TMN) histamine neurons and hydrolyzed. The resulting L-histidine may subsequently be converted into histamine, which could be responsible for the effects of carnosine on neurotransmission and hormone-like antiaging physiological function. The preliminary longitudinal studies of elderly individuals suggest that longer telomeres are associated with better survival, and an advanced oral nutritional support with nonhydrolyzed carnosine (or carcinine and patented compositions thereof) is a useful therapeutic tool for a critical TL maintenance that may fundamentally be applied in the treatment of age-related sight-threatening eye disorders, prolonged life expectancy, increased survival and chronological age of an organism in health control, smoking behavior, and disease. "Our pleasures were simple-they included survival." -Dwight D. Eisenhower, 34th President of the United States, 1953-1961.


Molecular characterization of a c-type lysozyme from the desert locust, Schistocerca gregaria (Orthoptera: Acrididae).
Lysozymes are bacteriolytic peptides that are implicated in the insect nonspecific innate immune responses. In this study, a full-length cDNA encoding a c-type lysozyme from Schistocerca gregaria (SgLys) has been cloned and characterized from the fat body of immune-challenged 5(th) instar. The deduced mature lysozyme is 119 amino acid residues in length, has a calculated molecular mass of 13.4 kDa and an isoelectric point (Ip) of 9.2. SgLys showed high identities with other insect lysozymes, ranging from 41.5% to 93.3% by BLASTp search in NCBI. Eukaryotic in vitro expression of the SgLys ORF (rSgLys) with an apparent molecular mass of ∼16 kDa under SDS-PAGE is close to the calculated molecular weight of the full-length protein. rSgLys displayed growth inhibitory activity against Gram-negative and Gram-positive bacteria. 3D structure modeling of SgLys, based on comparison with that of silkworm lysozyme, and sequence comparison with the helix-loop-helix (α-hairpin) structure of hen egg white lysozyme (HEWL) were employed to interpret the antibacterial potencies. Phylogenetic alignments indicate that SgLys aligns well with insect c-type lysozymes that expressed principally in fat body and hemocytes and whose role has been defined as immune-related. Western blot analysis showed that SgLys expression was highest at 6-12 h post-bacterial challenge and subsequently decreased with time. Transcriptional profiles of SgLys were determined by semi-quantitative RT-PCR analysis. SgLys transcript was upregulated at the highest level in fat body, hemocytes, salivary gland, thoracic muscles, and epidermal tissue. It was expressed in all developmental stages from egg to adult. These data indicate that SgLys is a predominant acute-phase protein that is expressed and upregulated upon immune challenge.


Prenatal developmental origins of behavior and mental health: The influence of maternal stress in pregnancy.
Accumulating research shows that prenatal exposure to maternal stress increases the risk for behavioral and mental health problems later in life. This review systematically analyzes the available human studies to identify harmful stressors, vulnerable periods during pregnancy, specificities in the outcome and biological correlates of the relation between maternal stress and offspring outcome. Effects of maternal stress on offspring neurodevelopment, cognitive development, negative affectivity, difficult temperament and psychiatric disorders are shown in numerous epidemiological and case-control studies. Offspring of both sexes are susceptible to prenatal stress but effects differ. There is not any specific vulnerable period of gestation; prenatal stress effects vary for different gestational ages possibly depending on the developmental stage of specific brain areas and circuits, stress system and immune system. Biological correlates in the prenatally stressed offspring are: aberrations in neurodevelopment, neurocognitive function, cerebral processing, functional and structural brain connectivity involving amygdalae and (pre)frontal cortex, changes in hypothalamo-pituitary-adrenal (HPA)-axis and autonomous nervous system.


Cost analysis in implementation studies of evidence-based practices for mental health and substance use disorders: a systematic review.
This study is a systematic literature review of cost analyses conducted within implementation studies on behavioral health services. Cost analysis of implementing evidence-based practices (EBP) has become important within implementation science and is critical for bridging the research to practice gap to improve access to quality healthcare services. Costing studies in this area are rare but necessary since cost can be a barrier to implementation and sustainment of EBP. We followed Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) methodology and applied the Consolidated Health Economic Evaluation Reporting Standards (CHEERS) checklist. Key search terms included: (1) economics, (2) implementation, (3) EBP, and (4) behavioral health. Terms were searched within article title and abstracts in: EconLit, SocINDEX, Medline, and PsychINFO. A total of 464 abstracts were screened independently by two authors and reduced to 37 articles using inclusion and exclusion criteria. After a full-text review, 18 articles were included. Findings were used to classify costs into direct implementation, direct services, and indirect implementation. While all studies included phases of implementation as part of their design, only five studies examined resources across multiple phases of an implementation framework. Most studies reported direct service costs associated with adopting a new practice, usually summarized as total EBP cost, cost per client, cost per clinician, and/or cost per agency. For studies with detailed analysis, there were eleven direct cost categories represented. For five studies that reported costs per child served, direct implementation costs varied from $886 to $9470 per child, while indirect implementation costs ranged from $897 to $3805 per child. This is the first systematic literature review to examine costs of implementing EBP in behavioral healthcare settings. Since 2000, 18 studies were identified that included a cost analysis. Given a wide variation in the study designs and economic methods, comparison across studies was challenging, which is a major limitation in the field, as it becomes difficult to replicate studies or to estimate future costs to inform policy decisions related to budgeting. We recommend future economic implementation studies to consider standard economic costing methods capturing costs across implementation framework phases to support comparisons and replicability.


Antibiotic research and development: business as usual?
The global burden of antibiotic resistance is tremendous and, without new anti-infective strategies, will continue to increase in the coming decades. Despite the growing need for new antibiotics, few pharmaceutical companies today retain active antibacterial drug discovery programmes. One reason is that it is scientifically challenging to discover new antibiotics that are active against the antibiotic-resistant bacteria of current clinical concern. However, the main hurdle is diminishing economic incentives. Increased global calls to minimize the overuse of antibiotics, the cost of meeting regulatory requirements and the low prices of currently marketed antibiotics are strong deterrents to antibacterial drug development programmes. New economic models that create incentives for the discovery of new antibiotics and yet reconcile these incentives with responsible antibiotic use are long overdue. DRIVE-AB is a €9.4 million public-private consortium, funded by the EU Innovative Medicines Initiative, that aims to define a standard for the responsible use of antibiotics and to develop, test and recommend new economic models to incentivize investment in producing new anti-infective agents.


Spinal arteriovenous malformation: evaluation of change in venous oxygenation with susceptibility-weighted MR imaging after treatment.
To evaluate the efficacy of susceptibility-weighted (SW) magnetic resonance (MR) imaging for the assessment of the posttreatment change in oxygen saturation in the draining vein in patients with spinal arteriovenous malformation (AVM). The study protocol was approved by the institutional review board, and written informed consent was obtained from all subjects. SW imaging was performed in 11 patients with spinal AVM before and after surgical or endovascular treatment. Eleven healthy subjects were included as a control group. A four-grade response scale was used for the visual assessment of the anterior spinal vein (ASV). For quantitative analysis, the phase value of the ASV was measured and oxygen saturation was calculated. Nonparametric multigroup comparison for visual assessment and one-way analysis of variance for quantitative measurement of oxygen saturation were used as statistical tests for comparison among three groups (pretreatment patients, posttreatment patients, and control subjects). Complete shunt occlusion in all patients was confirmed by using conventional angiography. For visual assessment, the average score of the pretreatment group was significantly less than that of the posttreatment and control groups. For quantitative analysis, the average oxygen saturation of the pretreatment group was significantly higher than that of the posttreatment and control groups, while no significant difference was observed between the posttreatment and control groups. After treatment, normalization of increased oxygen saturation was noninvasively observed by using SW imaging in patients with spinal AVM. SW imaging can be a useful tool for the assessment of treatment efficacy in patients with spinal AVM. (c) RSNA, 2010.


Costs of Transforming Established Primary Care Practices to Patient-Centered Medical Homes (PCMHs).
The patient-centered medical home (PCMH) shows promise for improving care and reducing costs. We sought to reduce the uncertainty regarding the time and cost of PCMH transformation by quantifying the direct costs of transforming 57 practices in a medical group to National Committee for Quality Assurance (NCQA)-recognized Level III PCMHs. We conducted structured interviews with corporate leaders, and with physicians, practice administrators, and office managers from a representative sample of practices regarding time spent on PCMH transformation and NCQA application, and related purchases. We then developed and sent a survey to all primary care practices (practice-level response rate: initial recognition-44.6%, renewal-35.7%). Direct costs were estimated as time spent multiplied by average hourly wage for the relevant job category, plus observed expenditures. We estimated HealthTexas' corporate costs for initial NCQA recognition (2010-2012) at $1,508,503; for renewal (2014-2016), $346,617; the Care Coordination resource costs an additional ongoing $390,790/year. A hypothetical 5-physician HealthTexas practice spent another estimated 239.5 hours ($10,669) obtaining, and 110.5 hours ($4,957) renewing, recognition. Centralized PCMH support reduces the burden on practices; however, overall time and cost remains substantial, and should be weighed against the mixed evidence regarding PCMH's impact on quality and costs of care.


Bubbles and Dust: Experimental Results of Dissolution Rates of Metal Salts and Glasses From Volcanic Ash Deposits in Terms of Surface Area, Chemistry, and Human Health Impacts.
Explosive volcanic eruptions lead to ash deposition and subsequent leaching of contaminants into soils or surface water, impacting flora and fauna, including human health. This study determined the control of ash surface area and chemical composition on ash dissolution rates. Fresh, unhydrated ash samples from four contrasting volcanoes were analyzed in the laboratory. Column leachate tests were used to compare leaching rates over a range of basaltic to andesitic ashes as a function of time and surface area, to analyze the effects of ash deposition. It was found that surface area, measured both geometrically and by multipoint Brunauer-Emmett-Teller analysis, generally increases for a short time, gradually decreases, then increases over the rest of the leaching experiment, due to area to mass ratio fluctuations. After the column leachate tests, postleaching water analyses for elemental compositions were conducted by inductively coupled plasma-mass spectrometry and ion chromatography. Steady state dissolution rates initially decayed rapidly due to the smallest size fraction of ash (dust), which provides a large area of fresh leachable surfaces as well as the rapid dissolution of highly soluble metal salts. Some of the dissolved concentrations of elements relevant to human and ecosystem health such as F, Cd, Se, As, and Cr rose above World Health Organization (WHO) drinking water standards within an hour of experimental leaching. In nature, however, safe consumption standards are further dependent upon bioaccumulation and chronic exposure. As such, individual and recurring ash deposition events have applications to emergency response and preparedness in volcanic regions.


Antimyeloma activity of two novel N-substituted and tetraflourinated thalidomide analogs.
Thalidomide alone or in combination with steroids has significant activity in multiple myeloma (MM). However, given its teratogenic potential, analogs have been synthesized, retaining the anti-MM activity without these side effects. We examined the anti-MM activity of two thalidomide analogs, CPS11 and CPS49. Direct cytotoxicity of the drugs on myeloma cell lines and patient myeloma cells was examined using thymidine uptake. Tumor cell apoptosis was evaluated by flow cytometry as well as Western blotting for caspase and PARP cleavage. Cellular signaling events were examined by immunoblotting for phosphorylated proteins. Both drugs inhibit proliferation of several MM cell lines sensitive and resistant to conventional therapies. They decrease secretion of IL-6, IGF, and VEGF by marrow stromal cells. Importantly, they inhibit proliferation of MM cells adherent to stromal cells. These drugs induce caspase-mediated apoptosis in MM cell lines, as well as patient MM cells. They inhibit the PI3K/Akt and JAK/STAT (signal transducers and activators of transcription) pathways in MM cells and are antiangiogenic in matrigel-based assays. CPS11 and CPS49 have potent antimyeloma activity and can overcome protective effects of the tumor microenvironment. They have potent antiangiogenic activity and direct effect on bone marrow stroma. These encouraging preclinical data provide the basis for further evaluation in the clinic.


Identification of a SNARE protein required for vacuolar protein transport in Schizosaccharomyces pombe.
Intracellular vesicle trafficking is mediated by a set of SNARE proteins in eukaryotic cells. Several SNARE proteins are required for vacuolar protein transport and vacuolar biogenesis in Saccharomyces cerevisiae. A search of the Schizosaccharomyces pombe genome database revealed a total of 17 SNARE-related genes. Although no homologs of Vam3p, Nyv1p, and Vam7p have been found in S. pombe, we identified one SNARE-like protein that is homologous to S. cerevisiae Pep12p. However, the disruptants transport vacuolar hydrolase CPY (SpCPY) to the vacuole normally, suggesting that the Pep12 homolog is not required for vacuolar protein transport in S. pombe cells. To identify the SNARE protein(s) involved in Golgi-to-vacuole protein transport, we have deleted four SNARE homolog genes in S. pombe. SpCPY was significantly missorted to the cell surface on deletion of one of the SNARE proteins, Fsv1p (SPAC6F12.03c), with no apparent S. cerevisiae ortholog. In addition, sporulation, endocytosis, and in vivo vacuolar fusion appear to be normal in fsv1Delta cells. These results showed that Fsv1p is mainly involved in vesicle-mediated protein transport between the Golgi and vacuole in S. pombe cells.


Chlamydia trachomatis major outer membrane protein variants escape neutralization by both monoclonal antibodies and human immune sera.
We have identified two families of novel Chlamydia trachomatis isolates with amino acid changes within the major outer membrane protein (MOMP) variable domains: one family of Da, D*, and D- and one family of Ia and I-. In order to determine whether these MOMP variants can escape antibody neutralization of infectivity, we tested both the D and I prototype strains and the variants in a complement-independent in vitro neutralization assay. We found that variants can indeed escape neutralization by both monoclonal antibodies and polyclonal human immune sera that neutralize the prototype strain.


Episomal maintenance of a bovine papilloma virus vector in transgenic mice.
We have used a bovine papillomavirus-based vector to generate transgenic mice. Transgenic mice result from either pronuclear or cytoplasmic injections of the vector into fertilized eggs. Of 30 mice generated by microinjection, 27 (90%) contained the vector in its episomal form, at less than one copy per cell. This represents a highly efficient means of gene transfer in which the transgene is in a controlled genetic environment.


Reactivation and/or growth of fecal coliform bacteria during centrifugal dewatering of anaerobically digested biosolids.
Fecal coliform bacteria are used as indicator organisms for the presence of pathogens. In sludges, it has often been assumed that the counts of fecal coliforms after digestion (where the sludges may also be called biosolids) are representative of the counts when the sludge is disposed or recycled, such as by land application. The possibility has been raised, however, that dewatering processes can lead to increased counts of fecal coliforms and, by inference, human pathogens. This paper presents data from previous studies of this possibility; the results were inconsistent but showed observable increases in fecal coliforms at one treatment plant. Additional studies were then performed at another treatment facility, which showed statistically significant increases in fecal coliform counts after dewatering and two days of aging. The increases exceeded two orders of magnitude and included two centrifuge types and two biosolids types. Artifacts of media type and enumeration method have been excluded, and shearing of the material by commercial blender did not produce the same effects.


Malaria: passive case detection and healthcare providers' choices of chemotherapy.
Prompt diagnosis and treatment play a central role in the malaria control programme in sub Saharan Africa. However, in most cases the diagnoses are never confirmed either for lack of facility or disinterest of healthcare providers resulting in over-diagnosis. To determine the proportion of clinically diagnosed malaria cases who could be confirmed with microscopy and evaluate compliance of healthcare providers with the National treatment guidelines for malaria. Participants were patients referred for malaria microscopy after the attending physicians had made clinical diagnosis of malaria. Thick blood smears were made under strict asepsis, stained and thereafter examined under oil immersion objective lens. Of the 630 patients who were referred with clinical impression of malaria, only 224 or 35.6% were positive for malaria parasite. The slide positive patients were younger with a mean age of 10.6 +/- 13.0 years versus 17.2 +/- 18.5 years [P < 0.005] for the slide negative individuals. There were only few instances of non-compliance with the National treatment guidelines for malaria. In conclusion, there appears to be over-diagnosis of malaria considering that only about a third of the clinical malaria cases were confirmed by microscopy. There is need for large epidemiological studies and possible policy review.


[Significative lengthening of the time of cornea conservation supporting its natural tension and curvature].
The success of cornea transplantations has always been a concern of world researchers to identify risk factors from the proper surgical techniques up to the several tissue storage procedures, such as the ones used in the new laboratories called eyes banks. However, the integral research works are still limited at present and they continue putting emphasis in the donor cornea storage that guarantee up to for this tissue feasibility. This research had the aim to contribute to the improvement of the corneal tissue quality taking into account the maintenance of its natural curvature and tensions combining them with some standard preservation solutions. This was achieved by means of a new mechanical system. Assays with rabbits were designed according to a sequential experimental groups design. In these ones, the study of the feasibility parameters behavior under different conservation media, related to the time, was splitted into four blocks. The first and second ones attended the cornea conservation in the short and intermediate-terms culture mediums as the ones at 4 degrees C degree isolated cornea preservation and the usual ones such as the Mc Carey Kaufman, Optisol and Likorol solutions used in the eyes banks. The third one was designed for the surgical assays coming from the two named groups and the last one focused the time study of these parameters for a new conservation technique, developed as an alternative medium to keep the cornea tissue curvatures and tensions as well as its nutrition and hydration. The results showed that tissue natural tensions maintenance is an significant factor in the conservation process, which involve the cornea feasibility for the its use in future transplantations. Since the new developed technique is very simple, this study is considered to be a significant contribution for the storage and biological material handling steps.


Underwater sound radiation from an elastically coated infinite plate with periodic inhomogeneities of finite width.
The underwater sound radiation from a line-force-driven coated infinite plate with periodically attached distributed inhomogeneities is investigated. A typical example of a distributed inhomogeneity is a signal conditioning plate (SCP), which can be inserted between the coating and a hydrophone to increase the strength of the incoming signal. Using the wavenumber transform approach, the surface normal velocity and radiated sound power of the plate are determined by solving the indexed equations of the wavenumber components. This study demonstrates that the elastic coating between the base plate and periodic inhomogeneities may reduce the effect of band-passes and band-gaps on the radiated sound power caused by the inhomogeneities. This is explained in detail by the insulation effect of the coating as a decoupler. The effect of the SCP's width and spatial period on the radiated sound is also examined. Finally, recommendations are given for the application of SCPs for underwater sensing purposes.


Tunable aggregation and luminescence of bis(diarylethene)sexithiophenes.
Diarylethenes with two different side groups (phenyl and chloro) were appended to both alpha-ends of a sexithiophene unit. The temperature dependent aggregation properties for both compounds were characterized by steady state and transient absorption spectroscopy. The peripheral side groups show an unexpectedly significant influence on the electronic properties of the sexithiophene core. Furthermore, the relative influence of the phenyl and chloro substituents on the aggregation behavior observed is remarkable. The phenyl compound exhibits formation of H-aggregates over a narrow temperature range, between 240 and 200 K, typical of strong intermolecular interactions. In contrast, the chloro compound shows gradual aggregation over a wide temperature range, forming H-aggregates albeit with weaker intermolecular interactions. The results demonstrate that minor changes in the structure lead to tunability of the aggregation and corresponding luminescence properties of sexithiophenes in solution and hold particular relevance to supramolecular and polymer systems based on sexithiophene units.


Prevention of deaths from harmful drinking in the United States: the potential effects of tax increases and advertising bans on young drinkers.
Harmful alcohol consumption is a leading cause of death in the United States. The majority of people who die from alcohol use begin drinking in their youth. In this study, we estimate the impact of interventions to reduce the prevalence of drinking among youth on subsequent drinking patterns and alcohol-attributable mortality. We first estimated the effect of public health interventions to decrease harmful drinking among youth from literature reviews and used life table methods to estimate alcohol-attributable years of life lost by age 80 years among the cohort of approximately 4 million U.S. residents aged 20 in the year 2000. Then, from national survey data on transitions in drinking habits by age, we modeled the impact of interventions on alcohol-attributable mortality. A tax increase and an advertising ban were the most effective interventions identified. In the absence of intervention, there would be 55,259 alcohol-attributable deaths over the lifetime of the cohort. A tax-based 17% increase in the price of alcohol of dollar 1 per six pack of beer could reduce deaths from harmful drinking by 1,490, equivalent to 31,130 discounted years of potential life saved or 3.3% of current alcohol-attributable mortality. A complete ban on alcohol advertising would reduce deaths from harmful drinking by 7,609 and result in a 16.4% decrease in alcohol-related life-years lost. A partial advertising ban would result in a 4% reduction in alcohol-related life-years lost. Interventions to prevent harmful drinking by youth can result in reductions in adult mortality. Among interventions shown to be successful in reducing youthful drinking prevalence, advertising bans appear to have the greatest potential for premature mortality reduction.


Multimode interference devices for focusing in microfluidic channels.
Low-cost, compact, automated optical microsystems for chemical analysis, such as microflow cytometers for identification of individual biological cells, require monolithically integrated microlenses for focusing in microfluidic channels, to enable high-resolution scattering and fluorescence measurements. The multimode interference device (MMI), which makes use of self-imaging in multimode waveguides, is shown to be a simple and effective alternative to the microlens for microflow cytometry. The MMIs have been designed, realized, and integrated with microfluidic channels in a silica-based glass waveguide material system. Focal spot sizes of 2.4 μm for MMIs have been measured at foci as far as 43.7 μm into the microfluidic channel.


Evaluation of the canine tympanic membrane by positive contrast ear canalography.
Positive contrast ear canalography was described briefly in 1973 as a method for detecting rupture of the tympanic membrane in dogs with otitis media. The purpose of this study was to evaluate the sensitivity and usefulness of the technique. The ears of 10 normal canine cadavers and 31 dogs with clinical signs of ear disease were examined using otoscopy, radiography and contrast radiography after infusing 2 to 5 ml of positive contrast medium into the ear canals. These examinations were repeated in the cadavers after the tympanic membrane had been punctured with a Spreull needle. In the cadavers 14 of 19 (74 per cent) of the tympanic membranes were visible otoscopically; contrast medium did not enter the tympanic bulla of any of the ears before the tympanic membrane was ruptured, but was visible in the bulla in every ear after rupture. In the clinical study, 40 of 61 (66 per cent) of the tympanic membranes were visible otoscopically, and 12 appeared to be ruptured. Radiographic signs of otitis media (increased opacity and/or thickening of the tympanic bulla) were identified in seven ears. Canalography was positive for rupture of the tympanic membrane in 13 ears, including four in which it appeared to be intact otoscopically. In normal canine ears, canalography was a more accurate method for detecting iatrogenic tympanic membrane rupture than otoscopy. In dogs with ear disease, canalography may be more sensitive for otitis media than either otoscopy or survey radiography.


Large-scale characterization of the macrolide resistome reveals high diversity and several new pathogen-associated genes.
Macrolides are broad-spectrum antibiotics used to treat a range of infections. Resistance to macrolides is often conferred by mobile resistance genes encoding Erm methyltransferases or Mph phosphotransferases. New erm and mph genes keep being discovered in clinical settings but their origins remain unknown, as is the type of macrolide resistance genes that will appear in the future. In this study, we used optimized hidden Markov models to characterize the macrolide resistome. Over 16 terabases of genomic and metagenomic data, representing a large taxonomic diversity (11 030 species) and diverse environments (1944 metagenomic samples), were searched for the presence of erm and mph genes. From this data, we predicted 28 340 macrolide resistance genes encoding 2892 unique protein sequences, which were clustered into 663 gene families (<70 % amino acid identity), of which 619 (94 %) were previously uncharacterized. This included six new resistance gene families, which were located on mobile genetic elements in pathogens. The function of ten predicted new resistance genes were experimentally validated in Escherichia coli using a growth assay. Among the ten tested genes, seven conferred increased resistance to erythromycin, with five genes additionally conferring increased resistance to azithromycin, showing that our models can be used to predict new functional resistance genes. Our analysis also showed that macrolide resistance genes have diverse origins and have transferred horizontally over large phylogenetic distances into human pathogens. This study expands the known macrolide resistome more than ten-fold, provides insights into its evolution, and demonstrates how computational screening can identify new resistance genes before they become a significant clinical problem.


The Contribution of Energy Systems in Repeated-Sprint Protocols: The Effect of Distance, Rest, and Repetition.
Purpose: This study aims to investigate the effects of rest intervals, sprint distance, and number of repetitions on performance variables, physiological responses, and energy system contributions in repeated-sprint protocols when total distance variable was the controlled (300 m). Method: Sixteen male soccer players participated in this study. The four protocols, each totaling a distance of 300 m, consisted of the combination of 15 and 30 m sprints with 30 s rest intervals (15meters30sec and 30meters30sec, respectively) and 1:5 work-rest ratios (15meters12sec and 30meters22sec, respectively). Aerobic, glycolytic, and phosphagen energy systems' contributions were calculated from the oxygen consumption (VO2) during the exercise, net lactate production, and the recovery VO2 kinetics using mono-exponential models. Repeated measures ANOVA with the Bonferroni correction was applied to examine the hypothesized differences. Results: The findings indicated that total sprint duration (F3:45=281.14; p<0.001), percentage of performance decline (F3:45=16.58; p<0.001), delta lactate (F3:45=39.72, p<0.001), rating of perceived exertion (F3:45=28.64; p<0.001), energy demand (F3:45=101.6; p<0.001), VO2 during the rest intervals (F3:45=42.72; p<0.001), and the absolute contribution of glycolytic (F3:45=119.6; p<0.001) and phosphagen energy systems (F3:45=72.9; p<0.001) were lowest in the 15meters30sec compared to other protocols. However, the relative contribution of aerobic system was greatest in the 15meters30sec compared to other protocols (F3:45=28.1, p<0.001). Both absolute (F3:45=119.6; p<0.001) and relative contribution of glycolytic system (F3:45=88.5, p<0.001) were greatest in the 30meters22sec compared to other protocols. Conclusion: This study showed that increasing sprint distance when rest interval is equal and decreasing rest interval when sprint distance is equal can increase the glycolytic system contribution.


Distinct time courses of renal protective action of angiotensin receptor antagonists and ACE inhibitors in chronic renal disease.
Although the angiotensin receptor antagonist (ARB) shares the angiotensin-II-blocking activity with the angiotensin-converting enzyme inhibitor (ACE-I), pharmacological mechanisms of action of these agents differ. We evaluated the temporal profiles of action of ACE-I and ARB on urinary protein excretion and nitrate/nitrate (NO(x)) excretion in hypertensive (140 and/or 90 mmHg) patients with chronic renal disease (serum creatinine < 265 (range, 44-265) micromol/l or creatinine clearance > 30 (range, 30-121) ml/min). Patients with mild (<1 g/day; range, 0.4-1.0) and moderate proteinuria (>1 g/day; range, 1.1-6.9) were randomly assigned to ACE-I- and ARB-treated groups, and were treated with ACE-I (trandolapril or perindopril) or ARB(losartan or candesartan) for 48 weeks. In all groups, treatment with ACE-I or ARB decreased blood pressure to the same level, but had no effect on creatinine clearance. In patients with mild proteinuria, neither ACE-I nor ARB altered urinary protein excretion. In patients with moderate proteinuria, ACE-I caused 44 +/- 6% reduction in proteinuria (from 2.7 +/- 0.5 to 1.5 +/- 0.4 g/day, n = 14) at 12 weeks, and this beneficial effect persisted throughout the protocol (48 weeks, 1.2 +/- 0.2 g/day). In contrast, ARB did not produce a significant decrease in proteinuria at 12 weeks (23 +/- 8%, n = 13), but a 41 +/- 6% reduction in proteinuria was observed at 48 weeks. Similarly, although early (12 weeks) increases in urinary NO(x) excretion were observed with ACE-I (from 257 +/- 70 to 1111 +/- 160 micromol/day) and ARB (from 280 +/- 82 to 723 +/- 86 micromol/day), the ARB-induced increase in NO(x) excretion was smaller than that by ACE-I (P < 0.05). In conclusion, although both ACE-I and ARB reduce blood pressure similarly, the effect of these agents on proteinuria differs in chronic renal disease with moderate proteinuria. Relatively early onset of the proteinuria-reducing effect was observed with ACE-I, which paralleled the increase in urinary NO(x) excretion. Conversely, ARB decreased proteinuria and increased urinary NO(x) excretion gradually. These time course-dependent changes in proteinuria and urinary NO(x) may reflect the pharmacological property of ACE-I and ARB, with regard to the action on bradykinin.


Cleaning Effectiveness of Three NiTi Rotary Instruments: A Focus on Biomaterial Properties.
Nickel-titanium (NiTi) instruments are commonly used for shaping the root canal system in endodontic practice. They are more flexible and have better cutting efficiency than conventional stainless steel files. The superelasticity of NiTi rotary files allows the clinicians to produce the desirable tapered root canal form with a reduced tendency to canal transportation and instrument fracture. HyFlex CM instruments are new NiTi rotary instruments with shape memory produced by an innovative methodology (patent pending) that uses a complex heating and cooling treatment that controls the material's memory. The aim of the present study was to compare the cleaning efficacy of two conventional (Mtwo, Revo-S) Ni-Ti rotary instruments with HyFlex CM. 30 single-rooted freshly extracted teeth were divided into three groups. Root canals were shaped with three NiTi instruments (Mtwo, Revo-S and HyFlex CM) using 5.25% NaOCl and 17% EDTA solutions. Specimens were fractured longitudinally and prepared for SEM analysis at standard magnification of 1000×. The presence/absence of debris smear layer and the presence/absence of smear layer at coronal, middle, and apical third of each canal were evaluated using a 5-step scale for scores. Numeric data were analyzed using Kruskall-Wallis and Mann-Whitney U statistical tests and significance was predetermined at P < 0.05. This study revealed significant differences among the various groups. Despite some minor differences, all instruments removed smear layer and debris produced during instrumentation. HyFlex CM seem to be not so effective in promoting cleanliness of root canal walls and in removing smear layer from dentine if compared to Mtwo and Revo-S.


Ecological adaptation during incipient speciation revealed by precise gene replacement.
To understand the role of adaptation in speciation, one must characterize the ecologically relevant phenotypic effects of naturally occurring alleles at loci potentially causing reproductive isolation. The desaturase2 gene of Drosophila melanogaster is such a locus. Two geographically differentiated ds2 alleles underlie a pheromonal difference between the Zimbabwe and Cosmopolitan races. We used a site-directed gene replacement technique to introduce an allele of ds2 from the Zimbabwe population into Cosmopolitan flies. We show that the Cosmopolitan allele confers resistance to cold as well as susceptibility to starvation when the entire genetic background is otherwise identical. We conclude that ecological adaptation likely accompanies sexual isolation between the two behavioral races of D. melanogaster.


Performance comparison of ferroelectric liquid-crystal-technology-based coherent optical multichannel correlators.
Our purpose is to compare two architectures when implemented with ferroelectric liquid-crystal technology: the conventional VanderLugt and joint transform correlators. The architectures are compared in the single-correlation and multichannel cases. The analysis covers both theoretical aspects and practical considerations regarding implementation. Specifications for a multichannel correlator design, including considerations of both spatial light modulators and architecture configurations, are discussed. Experimental results are presented for both architectures. Finally, the benefit resulting from extension to multichannel operation is discussed in terms of both multiplexing and algorithmic capabilities.


Automated Assessment of Cerebral Arterial Perforator Function on 7T MRI.
Blood flow velocity and pulsatility of small cerebral perforating arteries can be measured using 7T quantitative 2D phase contrast (PC) MRI. However, ghosting artifacts arising from subject movement and pulsating large arteries cause false positives when applying a previously published perforator detection method. To develop a robust, automated method to exclude perforators located in ghosting artifacts. Retrospective. Fifteen patients with vascular cognitive impairment or carotid occlusive disease and 10 healthy controls. 7T/cardiac-gated 2D PC MRI. Perforators were automatically excluded from ghosting regions, which were defined as bands in the phase-encoding direction of large arteries. As reference, perforators were manually excluded by two raters (T.A., J.J.M.Z.), based on perforator location with respect to visible ghosting artifacts. The performance of both censoring methods was assessed for the number of (Nincluded ), mean velocity (Vmean ), and pulsatility index (PI) of the included perforators. For within-method comparisons, inter- and intrarater reliability were assessed for the manual method, and test-retest reliability was assessed for both methods from repeated 2D PC scans (without repositioning). Intraclass correlation coefficients (ICCs) and their 95% confidence intervals (CIs) were determined for Nincluded , Vmean , and PI for all within-method comparisons. The ICC to compare between the two methods was determined with the use of both (test-retest) scans using a multilevel nonlinear mixed model. The automated censoring method showed a moderate to good ICC (95% CI) vs. manual censoring for Nincluded (0.73 [0.58-0.87]) and Vmean (0.90 [0.84-0.96]), and a moderate ICC for PI (0.57 [0.37-0.76]). The test-retest reliability of the manual censoring method was considerably lower than the interrater and intrarater reliability, indicating that scanner noise dominates the uncertainty of the analysis. The proposed automated censoring method can reliably exclude small perforators affected by ghosting artifacts. 3. 1.


Erythema nodosum as azathioprine hypersensitivity reaction in a patient with bullous pemphigoid.
A 65-year-old woman with bullous pemphigoid presented with fever and several red-purple nodular subcutaneous lesions on both lower legs 1 week after starting treatment with azathioprine (AZA). Biopsy of a skin nodule was compatible with erythema nodosum (EN) and hypersensitivity reaction to AZA was suspected. AZA was subsequently discontinued, observing complete remission of fever and EN within 2 weeks. This case highlights the importance of recognizing EN as a possible manifestation of hypersensitivity reaction to AZA.


[Video recording and data collection in the operating room: the way to a 'just culture' in the OR].
The Academic Medical Center, Amsterdam, has started a trial to evaluate the usefulness to team debriefings of performance reports generated by a medical data recorder (MDR) in the operating room (OR). Outcome performance reports in structured debriefings in a secure, non-punitive environment are likely to heighten the level of situational awareness of OR teams. This may prevent future error. In addition, the use of video and - even more likely - use of an MDR may contribute to establishing a 'just culture' in the OR. MDRs offer a wealth of data, but only if these data are processed well do the resulting outcome reports reveal insights useful for structured debriefings. The implementation of video recordings or MDRs must be preceded by carefully addressing privacy and litigation issues relating to both OR staff and patients. In this article, we address viewpoints and discuss implementation strategy and the legal considerations involved in enabling the use of video and data registration in the OR.


The development of a reinforcer choice assessment scale for persons with severe and profound mental retardation.
Reinforcement is a cornerstone for treating adaptive skills and eliminating problem behaviors for persons evincing mental retardation. To effectively use reinforcement, professionals must find reinforcers that are most effective for each individual. Establishing reinforcers is particularly important for persons functioning in the severe and profound ranges of mental retardation as reinforcers are often difficult to identify in this population. The current study aimed at developing a reinforcer rating scale that-would have applicability on a broad clinical basis, looking specifically at the psychometric properties of the scale.


Nonimmunoglobulin (non-Ig)/BCL6 gene fusion in diffuse large B-cell lymphoma results in worse prognosis than Ig/BCL6.
Chromosomal translocation involving the BCL6 gene affects not only immunoglobulin (Ig) genes but also a number of non-Ig genes as partners. The molecular anatomy of the BCL6 gene rearrangements in 39 cases with diffuse large B-cell lymphoma (DLBCL) by long-distance polymerase chain reaction-based assays was determined. The results showed that Ig genes were affected in 21 cases; non-Ig genes, 15 cases; a deletion of more than a 1-kb segment, 2 cases; and a point mutation, 1 case. Comparative studies between the 21 cases with Ig gene partners and the 17 cases with non-Ig gene partners, including 2 cases with the deletion, showed that the overall survival of the latter group of patients was significantly inferior to that of the former (P = .0440), and the estimated 2-year overall survival rates were 58.3% vs 17.6% (P = .005). Non-Ig/BCL6 fusion is a poor prognostic indicator of DLBCL, and DLBCL with BCL6 translocation could be subclassified according to the individual partner locus and/or gene. (Blood. 2000;96:2907-2909)


High scFv-receptor affinity does not enhance the antitumor activity of HER2-retargeted measles virus.
The relationship between ligand-receptor affinity and antitumor potency of an oncolytic virus was investigated using a panel of six HER2/neu (HER2)-targeted measles viruses (MVs) displaying single-chain antibodies (scFv) that bind to the same epitope on HER2, but with affinities ranging from 10(-6) to 10(-11) M. All viruses were able to infect SKOV3ip.1 human ovarian cancer cells in vitro, but only the high-affinity MV (Kd≥10(-8) M) induced cytopathic effects of syncytia formation in the cell monolayers. In contrast, all six viruses were therapeutically active in vivo against orthotopic human ovarian SKOV3ip.1 tumor xenografts in athymic mice compared with saline-treated controls. The oncolytic activities of MV displaying the high-affinity scFv (Kd=10(-9), 10(-10), 10(-11) M) were not significantly superior to MV displaying scFv with Kd of 10(-8) M or less. Results from this study suggest that increasing the receptor affinity of the attachment protein of an oncolytic MV has minimal impact on its in vivo efficacy against a tumor that expresses the targeted receptor.


Acute and chronic evolution of MRI findings in a case of posterior spinal cord ischemia.
Case report. Reveal the evolution of the magnetic resonance imaging (MRI) pattern in a patient with a posterior spinal artery infarction, which belongs to a subgroup of spinal cord ischemia syndromes and presents a rare cause of spinal cord injury. Our report underlines that diagnosis of spinal cord ischemia and thus clinical decision making remains challenging. University Hospital of Innsbruck and University Hospital of Salzburg, Austria. Here we present clinical, electrophysiological and imaging data in the acute, subacute and chronic phase of a woman who developed signs and symptoms related to a bilateral posterior spinal cord infarction. At the clinical nadir (24 h after symptom onset), MRI did not exhibit T2 hyperintensities. However, such MRI changes were detected 8 days after symptom onset and persisted until the latest follow-up at 5 months. Repeated MRI constitutes an indispensable diagnostic and follow-up tool for spinal cord ischemia. The imaging data in accordance with the electrophysiological measurements correlated well with the clinical presentation in the subacute und chronic phase. Therefore, further studies might allow using MRI following spinal cord ischemia as a prognostic marker for an individual outcome.


Inferring Rates and Length-Distributions of Indels Using Approximate Bayesian Computation.
The most common evolutionary events at the molecular level are single-base substitutions, as well as insertions and deletions (indels) of short DNA segments. A large body of research has been devoted to develop probabilistic substitution models and to infer their parameters using likelihood and Bayesian approaches. In contrast, relatively little has been done to model indel dynamics, probably due to the difficulty in writing explicit likelihood functions. Here, we contribute to the effort of modeling indel dynamics by presenting SpartaABC, an approximate Bayesian computation (ABC) approach to infer indel parameters from sequence data (either aligned or unaligned). SpartaABC circumvents the need to use an explicit likelihood function by extracting summary statistics from simulated sequences. First, summary statistics are extracted from the input sequence data. Second, SpartaABC samples indel parameters from a prior distribution and uses them to simulate sequences. Third, it computes summary statistics from the simulated sets of sequences. By computing a distance between the summary statistics extracted from the input and each simulation, SpartaABC can provide an approximation to the posterior distribution of indel parameters as well as point estimates. We study the performance of our methodology and show that it provides accurate estimates of indel parameters in simulations. We next demonstrate the utility of SpartaABC by studying the impact of alignment errors on the inference of positive selection. A C ++ program implementing SpartaABC is freely available in http://spartaabc.tau.ac.il.


Diabetes care quality: do large medical groups perform better?
To compare primary care management processes (CMPs) and outcome measures for diabetes quality among large, medium, and small medical groups. Observational comparison of differences in processes and outcomes over time among 329 primary care practices that agreed to participate and returned completed surveys in both 2017 and 2019. We used a standardized composite measure of diabetes quality along with its 5 components and a survey measure of the presence of systematic CMPs to compare the outcomes and processes of care among clinics that were in large (≥ 12 sites), medium (4-11 sites), and small (1-3 sites) medical groups. Practices from large groups had better performance than those in medium and small groups on the composite measure of diabetes outcomes in 2017 (46.5 vs 40.6 and 34.4, respectively; P < .001), as well as on each of the 5 component measures. They also had more CMPs in place (74.2% vs 66.9% and 61.4%; P < .001), including the 10 CMPs that are associated with the highest level of performance (84.2% vs 77.9% and 72.2%; P < .001). However, repeated measures in 2019 showed that the smaller groups had gained on both quality and CMP measures. There was also substantial overlap on both CMPs and performance among practices in groups of different sizes. On average, primary care practices that are part of large well-established medical groups outperformed smaller-sized groups in diabetes care quality, probably because they have the resources, leadership, and infrastructure to provide more consistent care through more organized CMPs.


Spliceosomal introns in the diplomonad parasite Giardia duodenalis revisited.
Complete reference genomes, including correct feature annotations, are a fundamental aspect of genomic biology. In the case of protozoan species such as Giardia duodenalis, a major human and animal parasite worldwide, accurate genome annotation can deepen our understanding of the evolution of parasitism and pathogenicity by identifying genes underlying key traits and clinically relevant cellular mechanisms, and by extension, the development of improved prevention strategies and treatments. This study used bioinformatics analyses of Giardia mRNA libraries to characterize known introns and identify new intron candidates, working towards completion of the G. duodenalis assemblage A strain 'WB' genome and further elucidating Giardia's gene expression. By using a set of experimentally validated positive control loci to calibrate our intron detection pipeline, we were able to detect evidence of previously missed candidate splice junctions directly from expressed transcript data. These intron candidates were further studied in silico using NMDS (non-metric multidimensional scaling) clustering to determine shared characteristics and their relative importance such as secondary structure, splicing efficiency and motif conservation, and thus to refine intron models. Results from this study identified 34 new intron candidates, with several potential introns showing evidence that secondary structure of the mRNA molecule might play a more significant role in splicing than previously reported eukaryotic splicing activity mediated by a reduced spliceosome present in G. duodenalis.


NR5A2 connects zygotic genome activation to the first lineage segregation in totipotent embryos.
Zygotic genome activation (ZGA) marks the beginning of the embryonic program for a totipotent embryo, which gives rise to the inner cell mass (ICM) where pluripotent epiblast arises, and extraembryonic trophectoderm. However, how ZGA is connected to the first lineage segregation in mammalian embryos remains elusive. Here, we investigated the role of nuclear receptor (NR) transcription factors (TFs), whose motifs are highly enriched and accessible from the 2-cell (2C) to 8-cell (8C) stages in mouse embryos. We found that NR5A2, an NR TF strongly induced upon ZGA, was required for this connection. Upon Nr5a2 knockdown or knockout, embryos developed beyond 2C normally with the zygotic genome largely activated. However, 4-8C-specific gene activation was substantially impaired and Nr5a2-deficient embryos subsequently arrested at the morula stage. Genome-wide chromatin binding analysis showed that NR5A2-bound cis-regulatory elements in both 2C and 8C embryos are strongly enriched for B1 elements where its binding motif is embedded. NR5A2 was not required for the global opening of its binding sites in 2C embryos but was essential to the opening of its 8C-specific binding sites. These 8C-specific, but not 2C-specific, binding sites are enriched near genes involved in blastocyst and stem cell regulation, and are often bound by master pluripotency TFs in blastocysts and embryonic stem cells (ESCs). Importantly, NR5A2 regulated key pluripotency genes Nanog and Pou5f1/Oct4, and primitive endoderm regulatory genes including Gata6 among many early ICM genes, as well as key trophectoderm regulatory genes including Tead4 and Gata3 at the 8C stage. By contrast, master pluripotency TFs NANOG, SOX2, and OCT4 targeted both early and late ICM genes in mouse ESCs. Taken together, these data identify NR5A2 as a key regulator in totipotent embryos that bridges ZGA to the first lineage segregation during mouse early development.