Objective Measurement of Brace Wear in Successfully Ponseti-Treated Clubfeet: Pattern of Decreasing Use in the First 2 Years.
Once Ponseti correction of a clubfoot is achieved and 3-month full-time bracing treatment is completed, part-time bracing treatment for 12 hours at night for 2 to 4 years is considered necessary to maintain a successful outcome. This study objectively documents the amount of daily orthosis wear time in those who maintained correction at age 2 years and, in so doing, determines how well patients' caretakers comply with the prescribed brace program. Patients <3 months old with idiopathic clubfeet when Ponseti treatment was initiated, who successfully maintained correction at age 2 years without surgery and who had complete objective brace wear data, were included. The foot abduction orthoses had a temperature data logger embedded in a shoe. Six 3-month time intervals were monitored in every patient as follows: full time: 0 to 3; night time: 4 to 6, 7 to 9, 10 to 12, 13 to 15, and 16 to 18 months. The families were not informed that hours of brace wear were being measured. One hundred twenty-four patients with 187 clubfeet were included. During the 0- to 3-month interval, wear time averaged 19.8 hr/d. After this period of full-time use, the night-time brace wear decreased over each of the subsequent five intervals: 11.9, 9.6, 8.6, 7.9, and 7.7 hours. By the 18-month period of brace wear, 1 of 3 patients wore the orthoses less than 6 hours per day, and nearly 1 of 2 patients wore the orthoses less than 8 hours per day. In patients evaluated at age 2 years whose clubfeet had successful nonsurgical treatment, night-time brace wear varied greatly and decreased over each 3-month period measured. By the second year of bracing treatment, nearly half of the patients wore them 8 hours or less. Level IV case series.


Effects of live yeast (Saccharomyces cerevisiae) and type of cereal on rumen microbial fermentation in a dual flow continuous culture fermentation system.
Live yeast additives may help optimize ruminal fermentation of high-grain diets, especially when the starch of the ration is highly fermentable. Eight dual flow continuous culture fermenters were used in a 2 × 2 factorial design in two replicated periods of 9 days (six for adaptation and three for sampling) to determine the effect of live yeast and type of cereal on rumen microbial fermentation and nutrient digestibility. Main factors were the addition of live yeast: no yeast (NY) vs. 2 × 107 CFU of yeast/g of diet (LY); and type of cereal in the diet: corn (CO) vs. barley (BA). All fermenters were fed 80 g dry matter/day of a 10-90 forage to concentrate diet and pH was allowed to fluctuate with an upper (6.6) and lower (5.5) limit. Treatment BA increased OM digestion, valerate proportion, peptides and ammonia N fractions, ammonia N flow, crude protein degradation and target copies of Megasphaera elsdenii; and decreased NDF digestion, propionate proportion, branched-chain VFA (BCVFA) concentration, AA-N fraction and nonammonia N flow. Treatment LY increased BCVFA and decreased ammonia N fraction and flow, and the target copies of Streptococcus bovis. Treatment LY decreased the slope of pH drop, the area under pH 6.0 and the gas production. These results suggest potential benefits of LY in stabilizing the fermentation of BA-based diets.


Saturation of cubic optical nonlinearity in long-chain polyene oligomers.
The scaling of the cubic nonlinearity gamma with chain length in polyenic molecules has received considerable theoretical attention. Earlier experimental investigations have been restricted to oligomers with fewer than 20 double bonds because of problems associated with the synthesis and solubility of conjugated molecules. These synthetic difficulties have been overcome in the present study by the use of modern living polymerization techniques. Solution measurements of gamma as a function of chain length in long-chain (up to 240 double bonds) model polyene oligomers are reported. A saturation of the increase of gamma with chain length is observed, and the onset of this saturation occurs for chain lengths considerably longer than predicted from theory.


Fibromyalgia, chronic fatigue syndrome, and myofascial pain syndrome.
The prevalence of chronic widespread pain in the general population in Israel was comparable with reports from the USA, UK, and Canada. Comorbidity with fibromyalgia (FM) resulted in somatic hyperalgesia in patients with irritable bowel syndrome. One sixth of the subjects with chronic widespread pain in the general population were also found to have a mental disorder. Mechanisms involved in referred pain, temporal summation, muscle hyperalgesia, and muscle pain at rest were attenuated by the N-methyl-D-aspartate (NMDA) antagonist, ketamine, in FM patients. Delayed corticotropin release, after interleukin-6 administration, in FM was shown to be consistent with a defect in hypothalamic corticotropin-releasing hormone neural function. The basal autonomic state of FM patients was characterized by increased sympathetic and decreased parasympathetic systems tones. The severity of functional impairment as assessed by the Medical Outcome Survey Short Form (SF-36) discriminated between patients with widespread pain alone and FM patients. Chronic fatigue syndrome (CFS) occurred in about 0.42% of a random community-based sample of 28,673 adults in Chicago, Illinois. A significant clinical overlap between CFS and FM was reported. Cytokine dysregulation was not found to be a singular or dominant factor in the pathogenesis of CFS. A favorable outcome of CFS in children was reported; two thirds recovered and resumed normal activities. No major therapeutic trials in FM and CFS were reported over the past year.


Space-dependent color gamut mapping: a variational approach.
Gamut mapping deals with the need to adjust a color image to fit into the constrained color gamut of a given rendering medium. A typical use for this tool is the reproduction of a color image prior to its printing, such that it exploits best the given printer/medium color gamut, namely the colors the printer can produce on the given medium. Most of the classical gamut mapping methods involve a pixel-by-pixel mapping and ignore the spatial color configuration. Recently proposed spatial-dependent approaches for gamut mapping are either based on heuristic assumptions or involve a high computational cost. In this paper, we present a new variational approach for space-dependent gamut mapping. Our treatment starts with the presentation of a new measure for the problem, closely related to a recent measure proposed for Retinex. We also link our method to recent measures that attempt to couple spectral and spatial perceptual measures. It is shown that the gamut mapping problem leads to a quadratic programming formulation, guaranteed to have a unique solution if the gamut of the target device is convex. An efficient numerical solution is proposed with promising results.


Genetic Characterisation of Plasmodium falciparum Isolates with Deletion of the pfhrp2 and/or pfhrp3 Genes in Colombia: The Amazon Region, a Challenge for Malaria Diagnosis and Control.
Most Plasmodium falciparum-detecting rapid diagnostic tests (RDTs) target histidine-rich protein 2 (PfHRP2). However, P. falciparum isolates with deletion of the pfhrp2 gene and its homolog gene, pfhrp3, have been detected. We carried out an extensive investigation on 365 P. falciparum dried blood samples collected from seven P. falciparum endemic sites in Colombia between 2003 and 2012 to genetically characterise and geographically map pfhrp2- and/or pfhrp3-negative P. falciparum parasites in the country. We found a high proportion of pfhrp2-negative parasites only in Amazonas (15/39; 38.5%), and these parasites were also pfhrp3-negative. These parasites were collected between 2008 and 2009 in Amazonas, while pfhrp3-negative parasites (157/365, 43%) were found in all the sites and from each of the sample collection years evaluated (2003 to 2012). We also found that all pfhrp2- and/or pfhrp3-negative parasites were also negative for one or both flanking genes. Six sub-population clusters were established with 93.3% (14/15) of the pfhrp2-negative parasites grouped in the same cluster and sharing the same haplotype. This haplotype corresponded with the genetic lineage BV1, a multidrug resistant strain that caused two outbreaks reported in Peru between 2010 and 2013. We found this BV1 lineage in the Colombian Amazon as early as 2006. Two new clonal lineages were identified in these parasites from Colombia: the genetic lineages EV1 and F. PfHRP2 sequence analysis revealed high genetic diversity at the amino acid level, with 17 unique sequences identified among 53 PfHRP2 sequences analysed. The use of PfHRP2-based RDTs is not recommended in Amazonas because of the high proportion of parasites with pfhrp2 deletion (38.5%), and implementation of new strategies for malaria diagnosis and control in Amazonas must be prioritised. Moreover, studies to monitor and genetically characterise pfhrp2-negative P. falciparum parasites in the Americas are warranted, given the extensive human migration occurring in the region.


Fiber-based tunable microcavity fluidic dye laser.
We investigate a tunable fluidic dye laser formed by a microcavity filled with a dye solution. We achieve a wide 18 nm tunability of the laser wavelength by controlling the cavity length for the first time. The microcavity is made of a silica capillary and two aligned fibers with end faces Au-coated. The Rhodamine 6G dye solution flowing through the microcavity is pumped by 532 nm wavelength laser pulses. Laser emission around 570 nm in the form of TE mode with a threshold of about 58 μJ/pulse is obtained. This work suggests a fiber-based convenient approach to achieve wavelength tunability and integration with lab-on-a-chip systems.


[Clinical evaluation on differential quantitation of human chorionic gonadotropin in testicular cancer].
In 113 patients with testicular germ cell tumor, the authors determined human chorionic gonadotropin (hCG) and the free beta subunit levels in sera by differential quantitation using homologous hCG-beta radioimmunoassay (RIA) and hCG enzyme immunoassay (EIA), respectively. In 59 patients with seminoma, intact hCG was positive in 6 patients (10.1%), whereas free hCG-beta was positive in 23 patients (39.0%). Syncytiotrophoblastic giant cells were detected in 4 cases out of 6 patients with positive intact hCG seminoma. Seventeen cases (28.8%) revealed positive free hCG-beta only without elevation of intact hCG. In 54 patients with nonseminoma, both intact hCG and free hCG-beta were positive in 36 cases (66%). In some cases, hCG-beta/hCG ratio increased up to 275% when recurrence of tumor developed. Molecular heterogeneity of hCG is closely related to proliferation and differentiation of hCG producing cells. The differential quantitation of intact hCG and hCG-beta using these two assays is valuable for clinical detection of molecular heterogeneity of hCG in testicular tumor.


Atypical interference control in children with AD/HD with elevated theta/beta ratio.
The theta/beta ratio (TBR) is a major area of interest within electroencephalogram (EEG) research in AD/HD. While researchers suggest a prognostic role for TBR in AD/HD, its relationship to behavior remains uncertain. Recent evidence suggests that elevated TBR in AD/HD may be related to atypical inhibition, particularly at an attentional level. This study aimed to examine the performance on three inhibitory tasks of children with AD/HD. Fifty-eight children with AD/HD participated, divided into an elevated TBR (ET) group and a control group (CT). A behavioral disassociation was found - compared to CT, ET showed more difficulty in inhibiting surrounding stimuli but had less day-to-day inhibitory issues measured by BRIEF. There was no significant group difference on response inhibition. The results support the prognostic value of TBR in AD/HD. Elevated TBR may be an inhibitory biomarker; further studies are needed to explore the behavioral implications in patients without elevated TBR.


Postoperative analgesia with epidural opioids after cesarean section: Comparison of sufentanil, morphine and sufentanil-morphine combination.
Epidural analgesia with opioid provides good control of postoperative pain in cesarean section, thereby improving the mother's ability to mobilize and interact with her newborn infant. The aim of this study is to evaluate and compare the analgesic actions and side effects of epidural analgesia with sufentanil, morphine or combination of the two after cesarean section. 60 women undergoing elective cesarean section were allocated into three groups of 20 each in a randomized blinded fashion. Epidural analgesia was administered with sufentanil 50 mcg in Group S; morphine 4 mg in Group M; and, a combination of sufentanil 25 mcg and morphine 2 mg was used in Group SM. Analgesic efficacy in terms of onset of action and duration of analgesia was assessed by using the Visual Analog Scale (0 to 10 cm) for 24 hours. Number of opioid doses needed in 24 hours was noted. Side effects like respiratory depression /excessive sedation, pruritus and nausea were recorded. Onset of action were at 7.6 ± 1.5 minutes in group S, 67.6 ± 1.5 minutes in group M and 12.2 ± 2.6 minutes in group SM. Duration of analgesia was longer in group M 17.5 ± 1.9 hours and SM 13.8 ± 1.6 hours than in group S 5.2 ± 1.2 hours. More doses of analgesia were required in group S compared to group M and SM. Side effects were comparable in the three groups. Epidural administration of a combination of sufentanil and morphine offered the advantage of faster onset of action and longer duration of analgesia as compared to the two drugs administered alone.


Stabilization of clinical samples collected for quantitative bioanalysis--a reflection from the European Bioanalysis Forum.
In bioanalysis of small molecules, the analyte concentration in the measured samples should reflect the concentration during sample collection. Precautions may be needed to prevent over- or under-estimation of the obtained result. This might require the addition of stabilizers to prevent degradation or nonspecific binding. For unstable drugs, it is essential to know how analytes can be stabilized before the start of the clinical study. Although the stabilization methods are well documented, the impact of the stabilization on the clinical workflow is not properly addressed. Already during method development, the clinical implications in terms of personnel safety, ease of use, training possibilities and staff capacity should be taken into account, and validation of the bioanalytical method should reflect collection procedures.


Multimodality molecular imaging of glioblastoma growth inhibition with vasculature-targeting fusion toxin VEGF121/rGel.
Vascular endothelial growth factor A (VEGF-A) and its receptors, Flt-1/FLT-1 (VEGFR-1) and Flk-1/KDR (VEGFR-2), are key regulators of tumor angiogenesis and tumor growth. The purpose of this study was to determine the antiangiogenic and antitumor efficacies of a vasculature-targeting fusion toxin (VEGF(121)/rGel) composed of the VEGF-A isoform VEGF(121) linked with a G(4)S tether to recombinant plant toxin gelonin (rGel) in an orthotopic glioblastoma mouse model by use of noninvasive in vivo bioluminescence imaging (BLI), MRI, and PET. Tumor-bearing mice were randomized into 2 groups and balanced according to BLI and MRI signals. PET with (64)Cu-1,4,7,10-tetraazacyclododedane-N,N',N'',N'''-tetraacetic acid (DOTA)-VEGF(121)/rGel was performed before VEGF(121)/rGel treatment. (18)F-Fluorothymidine ((18)F-FLT) scans were obtained before and after treatment to evaluate VEGF(121)/rGel therapeutic efficacy. In vivo results were confirmed with ex vivo histologic and immunohistochemical analyses. Logarithmic transformation of peak BLI tumor signal intensity revealed a strong correlation with MRI tumor volume (r = 0.89, n = 14). PET with (64)Cu-DOTA-VEGF(121)/rGel before treatment revealed a tumor accumulation (mean +/- SD) of 11.8 +/- 2.3 percentage injected dose per gram at 18 h after injection, and the receptor specificity of the tumor accumulation was confirmed by successful blocking of the uptake in the presence of an excess amount of VEGF(121). PET with (18)F-FLT revealed significant a decrease in tumor proliferation in VEGF(121)/rGel-treated mice compared with control mice. Histologic analysis revealed specific tumor neovasculature damage after treatment with 4 doses of VEGF(121)/rGel; this damage was accompanied by a significant decrease in peak BLI tumor signal intensity. The results of this study suggest that future clinical multimodality imaging and therapy with VEGF(121)/rGel may provide an effective means to prospectively identify patients who will benefit from VEGF(121)/rGel therapy and then stratify, personalize, and monitor treatment to obtain optimal survival outcomes.


Molecular mechanisms of Ebola pathogenesis.
Ebola viruses (EBOVs) and Marburg viruses (MARVs) are among the deadliest human viruses, as highlighted by the recent and widespread Ebola virus outbreak in West Africa, which was the largest and longest epidemic of Ebola virus disease (EVD) in history, resulting in significant loss of life and disruptions across multiple continents. Although the number of cases has nearly reached its nadir, a recent cluster of 5 cases in Guinea on March 17, 2016, has extended the enhanced surveillance period to June 15, 2016. New, enhanced 90-d surveillance windows replaced the 42-d surveillance window to ensure the rapid detection of new cases that may arise from a missed transmission chain, reintroduction from an animal reservoir, or more important, reemergence of the virus that has persisted in an EVD survivor. In this review, we summarize our current understanding of EBOV pathogenesis, describe vaccine and therapeutic candidates in clinical trials, and discuss mechanisms of viral persistence and long-term health sequelae for EVD survivors.


Discovery of the antibiotic phosacetamycin via a new mass spectrometry-based method for phosphonic acid detection.
Naturally occurring phosphonates such as phosphinothricin (Glufosinate, a commercially used herbicide) and fosfomycin (Monurol, a clinically used antibiotic) have proved to be potent and useful biocides. Yet this class of natural products is still an under explored family of secondary metabolites. Discovery of the biosynthetic pathways responsible for the production of these compounds has been simplified by using gene based screening approaches, but detection and identification of the natural products the genes produce have been hampered by a lack of high-throughput methods for screening potential producers under various culture conditions. Here, we present an efficient mass-spectrometric method for the selective detection of natural products containing phosphonate and phosphinate functional groups. We have used this method to identify a new phosphonate metabolite, phosacetamycin, whose structure, biological activity, and biosynthetic gene cluster are reported.


Trace elements in health and diseases.
Remarkable progress has occurred in many branches of biology and health sciences during the last few decades. Trace element (TE) research has definitely shared in this explosion of scientific knowledge. Due to the improvements in analytical technology, the discovery of TEs in organism was realized. The developments of TE research have been promoted by the demand of public health (e.g. deficiency or toxicity). The profound knowledge of nutritional importance of TEs has been achieved with application of advanced methods in biological, medical and chemical fields, etc. In this paper, a new definition of essentiality of TEs is introduced. According to this definition, only ten TEs (Fe, Zn, F, Cu, I, Se, Mn, Mo, Cr, Co) are considered to be essential to humans. The others need more evidence to prove their essentiality in humans. The recent progress on the biochemical and immunological functions of TEs and on the roles of TEs in brain development are briefly described. The TEs, mainly Se, I, Zn, Fe, are more closely related to public health. Also, emphases are laid on balancing all nutrients when new knowledge of essential TEs is applied in public health.


Surgical treatment and complications of treating pancreatic tumor.
Pancreatic tumor is one with the worst prognosis of all cancers, and the tenth most frequent cancer in Europe, making the 3% of all cancers affecting both sexes. Most patients seek treatment when the disease is in its advanced stage and the level for possible resectability is low. Late presentation of the disease is responsible for the short survival period of 6 months and a five-year survival of 0.4 to 5% of patients. At the Clinic for Surgery in Tuzla during period from January 1st 1996, to January 1st 2011, a total of 127 resection surgeries were performed due to malignant tumors. The goal of this study was to show that adequate assessment of operability, proper surgical strategy and modern techniques of creating anastomoses reduces morbidity and mortality, results in fewer postoperative complications and contributes to better surgical results. In our study sample the most common place of tumor location was the head of pancreas, in 69 (59.7%) patients. Men develop this type of cancer more often than women in the ratio of 2:1, while the median age of patients was 62 years. We faced postoperative complications in 37 (29.1%) patients, pancreatic fistula being the most prevalent complication, occurring in 16 (12.6%) patients. Overall early and late postoperative mortality was observed in 12 (9.8%) patients. Patients with chronic and hereditary pancreatitis are at a higher risk for developing pancreatic cancer and should be screened for the purpose of early diagnosis. The staging of pancreatic cancer has improved, with the accuracy of 85-90%. Postoperative complications, morbidity, and mortality are significantly reduced (p < 0.05) if the standardized operational procedure is applied and if modern techniques are used to create pancreaticojejunal anastomosis as the anastomosis carrying the highest risk.


Interaction of pituitary hormones and expression of clock genes modulated by bone morphogenetic protein-4 and melatonin.
Functional interaction of clock genes and pituitary hormones was investigated by focusing on bone morphogenetic protein (BMP)-4 and melatonin actions in anterior pituitary cells. A significant correlation between the mRNA expression of proopiomelanocortin (POMC) and Per2 was revealed in serial cultures of corticotrope AtT20 cells. Knockdown of Per2 expression by siRNA in AtT20 cells resulted in a significant reduction of POMC mRNA level with or without corticotropin-releasing hormone (CRH) stimulation. Treatments with BMP-4 and melatonin, both of which suppress POMC expression, reduced Per2 mRNA as well as protein levels in AtT20 cells. On the other hand, in lactosomatotrope GH3 cells, an expressional correlation was found between prolactin (PRL) and Clock mRNA levels, which was attenuated in the presence of forskolin treatment. The siRNA-mediated knockdown of Clock expression, but not that of Bmal1, significantly reduced PRL mRNA levels in GH3 cells. Interestingly, Clock mRNA and protein levels did not fluctuate with melatonin, BMP-4 or forskolin treatment, although Bmal1 expression was significantly increased by forskolin treatment. Collectively, a significant correlation between the expression of POMC and Per2 and that between PRL and Clock were uncovered in corticotrope and lactosomatotrope cells, respectively. Per2 expression was inhibited by POMC modulators including melatonin and BMP-4, while Clock expression was steadily maintained. Thus, the effects of melatonin and BMP-4 on clock gene expression may imply differential stability of circadian rhythms of adrenocorticotropin (ACTH) and PRL secreted from the anterior pituitary.


Regulatory effects of sestrin 3 (SESN3) in BCR-ABL expressing cells.
Chronic myeloid leukemia (CML) and Ph+ acute lymphoblastic leukemia (ALL) are characterized by the presence of the BCR-ABL oncoprotein, which leads to activation of a plethora of pro-mitogenic and pro-survival pathways, including the mTOR signaling cascade. We provide evidence that in BCR-ABL expressing cells, treatment with tyrosine kinase inhibitors (TKIs) results in upregulation of mRNA levels and protein expression of sestrin3 (SESN3), a unique cellular inhibitor of mTOR complex 1 (mTORC1). Such upregulation appears to be mediated by regulatory effects on mTOR, as catalytic inhibition of the mTOR kinase also induces SESN3. Catalytic mTOR inhibition also results in upregulation of SESN3 expression in cells harboring the TKI-insensitive T315I-BCR-ABL mutant, which is resistant to imatinib mesylate. Overexpression of SESN3 results in inhibitory effects on different Ph+ leukemic cell lines including KT-1-derived leukemic precursors, indicating that SESN3 mediates anti-leukemic responses in Ph+ cells. Altogether, our findings suggest the existence of a novel mechanism for the generation of antileukemic responses in CML cells, involving upregulation of SESN3 expression.


Publication rate and citation counts for preprints released during the COVID-19 pandemic: the good, the bad and the ugly.
Preprints are preliminary reports that have not been peer-reviewed. In December 2019, a novel coronavirus appeared in China, and since then, scientific production, including preprints, has drastically increased. In this study, we intend to evaluate how often preprints about COVID-19 were published in scholarly journals and cited. We searched the iSearch COVID-19 portfolio to identify all preprints related to COVID-19 posted on bioRxiv, medRxiv, and Research Square from January 1, 2020, to May 31, 2020. We used a custom-designed program to obtain metadata using the Crossref public API. After that, we determined the publication rate and made comparisons based on citation counts using non-parametric methods. Also, we compared the publication rate, citation counts, and time interval from posting on a preprint server to publication in a scholarly journal among the three different preprint servers. Our sample included 5,061 preprints, out of which 288 were published in scholarly journals and 4,773 remained unpublished (publication rate of 5.7%). We found that articles published in scholarly journals had a significantly higher total citation count than unpublished preprints within our sample (p < 0.001), and that preprints that were eventually published had a higher citation count as preprints when compared to unpublished preprints (p < 0.001). As well, we found that published preprints had a significantly higher citation count after publication in a scholarly journal compared to as a preprint (p < 0.001). Our results also show that medRxiv had the highest publication rate, while bioRxiv had the highest citation count and shortest time interval from posting on a preprint server to publication in a scholarly journal. We found a remarkably low publication rate for preprints within our sample, despite accelerated time to publication by multiple scholarly journals. These findings could be partially attributed to the unprecedented surge in scientific production observed during the COVID-19 pandemic, which might saturate reviewing and editing processes in scholarly journals. However, our findings show that preprints had a significantly lower scientific impact, which might suggest that some preprints have lower quality and will not be able to endure peer-reviewing processes to be published in a peer-reviewed journal.


[Association between wound size after open cholecystectomy with severity of iatrogenic biliary tract lesions].
Transoperative biliary tract injury during open or laparoscopic cholecystectomy is a catastrophic event associated with significant morbidity and mortality. Our objective was to determine if wound size during open cholecystectomy is associated with more complex biliary tract injuries. Prospective cohort study performed between March 2006 and February 2007. Sixty-six patients with biliary tract injuries after open cholecystectomy were included. Wound size was analyzed. Sixty six patients were included, 70% were female with a median age of 44. Seventy four percent were treated for acute cholecystitis. Most participants were overweight or had various degrees of obesity. Biliary tract injuries were not recognized during the primary surgical procedure in 76% of cases. All patients with severe biliary tract injuries (Strasberg E-3 and E-4) had a wound size less than 10 cm in length. Wound size was associated with unrecognized injuries (p=0.000), as well as with injury severity (p=0.000). We were notable to demonstrate a statistically significant association between biliary tract injuries and elective or emergency surgical procedures and midline or subcostal surgical incision for cholecystectomy. Our findings suggest that minor surgical access during open cholecystectomy may constitute a risk factor since smaller incisions were associated with more severe biliary tract injuries and an inability to observe this damage during the surgical procedure. We suggest to adhere strictly to the guidelines of an adequate surgical exposure during open cholecystectomy to prevent biliary tract injuries.


Beliefs, barriers, social support, and environmental influences related to diabetes risk behaviours among women with a history of gestational diabetes.
Women with previous gestational diabetes mellitus (GDM) are at increased risk of developing type 2 diabetes; this risk is higher in non-Caucasian women. This study explored the beliefs, attitudes, social support, environmental influences and other factors related to diabetes risk behaviours among Arabic, Cantonese/Mandarin, and English speaking women with recent GDM. Women living in the Sydney metropolitan area (Australia) who had GDM 6-36 months previously were included. In-depth semi-structured telephone interviews on women's experiences and perceptions of GDM and the lifestyle risk factors for developing type 2 diabetes were conducted in the language participants spoke at home (n=20 Arabic, 20 Cantonese/Mandarin, 17 English). Data were analysed for underlying themes using NVivo software. Mental distress, role perceptions, social support and cultural expectations were major issues related to women's struggles to find the right balance between the large proportion of household and child care responsibilities and leading a healthy lifestyle. Women's ability to follow a healthy lifestyle is embedded in their psychological wellbeing and the social and cultural context of their lives. The study highlights the need for a holistic approach that ensures personal support and access to services as well as lifestyle specific programs.


The deposition of metal nanoparticles on carbon surfaces: the role of specific functional groups.
The enormous complexity of a typical heterogeneous catalyst makes understanding the development and properties of any active nanoparticles present extremely challenging. In the case of carbon based catalysts that difficulty is compounded by the variability of the carbon powders used. We have previously developed a strategy that addresses these problems by mimicking the catalyst preparation conditions very closely but using highly ordered pyrolytic graphite crystals (HOPG) as a model surface. This enables us to examine the effects of specific functional groups on nanoparticle formation. We report here an extension of our work characterising functional groups on the HOPG surface, using XPS and AFM to explore the deposition of gold from aqueous solution onto HOPG surfaces treated in a variety of ways to alter the surface functionality. The structure and oxidation state of the resulting nanoparticles depend critically on the nature of the functional groups present and offers some insight into the development of catalysts based on these materials. Hydroxyls are identified as key functional species, reducing gold ions to their metallic state whilst being oxidised themselves to carbonyls. Carbonyls meanwhile promote the nucleation of Au3+, creating a network of islands at the HOPG surface. The results have relevance not only to catalysts using activated carbons but also the new generation of materials based on graphene and carbon nanotubes.


Infertility due to defective sperm flagella caused by an intronic deletion in DNAH17 that perturbs splicing.
Artificial insemination in pig (Sus scrofa domesticus) breeding involves the evaluation of the semen quality of breeding boars. Ejaculates that fulfill predefined quality requirements are processed, diluted and used for inseminations. Within short time, eight Swiss Large White boars producing immotile sperm that had multiple morphological abnormalities of the sperm flagella were noticed at a semen collection center. The eight boars were inbred on a common ancestor suggesting that the novel sperm flagella defect is a recessive trait. Transmission electron microscopy cross-sections revealed that the immotile sperm had disorganized flagellar axonemes. Haplotype-based association testing involving microarray-derived genotypes at 41,094 SNPs of six affected and 100 fertile boars yielded strong association (P = 4.22 × 10-15) at chromosome 12. Autozygosity mapping enabled us to pinpoint the causal mutation on a 1.11 Mb haplotype located between 3,473,632 and 4,587,759 bp. The haplotype carries an intronic 13-bp deletion (Chr12:3,556,401-3,556,414 bp) that is compatible with recessive inheritance. The 13-bp deletion excises the polypyrimidine tract upstream exon 56 of DNAH17 (XM_021066525.1: c.8510-17_8510-5del) encoding dynein axonemal heavy chain 17. Transcriptome analysis of the testis of two affected boars revealed that the loss of the polypyrimidine tract causes exon skipping which results in the in-frame loss of 89 amino acids from DNAH17. Disruption of DNAH17 impairs the assembly of the flagellar axoneme and manifests in multiple morphological abnormalities of the sperm flagella. Direct gene testing may now be implemented to monitor the defective allele in the Swiss Large White population and prevent the frequent manifestation of a sterilizing sperm tail disorder in breeding boars.


Kinetic study of the aspartate/glutamate carrier in intact rat heart mitochondria and comparison with a reconstituted system.
The homologous exchange of external [14C] aspartate/internal aspartate catalyzed by the aspartate/glutamate carrier of rat heart mitochondria was investigated using aspartate-loaded, glutamate-depleted mitochondria. An inhibitor-stop technique was developed for kinetic studies by applying pyridoxal phosphate. Direct initial rate determinations from the linear phase of [14C] aspartate uptake were insufficiently accurate at high external and/or low internal substrate concentrations. Therefore, the full time-course of [14C] aspartate uptake until reaching isotope equilibrium was fitted by a single exponential function and was used to calculate reliable initial steady-state rates. This method was applied in bisubstrate analyses of the antiport reaction for different external and internal aspartate concentrations. The kinetic patterns obtained in double reciprocal plots showed straight lines converging on the abscissa. This result is consistent with a sequential antiport mechanism. It implies the existence of a catalytic ternary complex that is formed by the translocator and substrate molecules bound from both sides of the membrane. The Km values for aspartate were clearly different for the external and the internal sides of the membrane, 216 +/- 23 microM and 2.4 +/- 0.5 mM, respectively. These values indicated a definite transmembrane asymmetry of the carrier. The same asymmetry became evident when investigating the isolated protein from bovine heart mitochondria after reconstitution into liposomes. In this case the Km values for external and internal aspartate were determined to be 123 +/- 11 microM and 2.8 +/- 0.6 mM, respectively. This comparison demonstrates a right-side out orientation of the carrier after insertion into liposomal membranes. The sequential transport mechanism of the aspartate/glutamate carrier, elucidated both in proteoliposomes and in mitochondria, also seems to be a common characteristic of other mitochondrial antiport carriers.


In Vitro Eradication of Planktonic, Saliva and Biofilm Bacteria Using Lingonberry Extract as a Photosensitizer for Visible Light Plus Water-Filtered Infrared-A Irradiation.
Antimicrobial photodynamic treatment (aPDT) with visible light plus water-filtered infrared-A irradiation (VIS-wIRA) and natural single- or multi-component photosensitizers (PSs) was shown to have potent antimicrobial activity. The aim of this study was to obtain information on the antimicrobial effects of aPDT-VIS-wIRA with lingonberry extract (LE) against bacteria that play a role in oral health. Planktonic bacterial cultures of the Gram-positive E. faecalis T9, S. mutans DSM20523, S. oralis ATCC 35037 and S. sobrinus PSM 203513, the Gram-negative N. oralis 14F2 FG-15-7B, F. nucleatum ATCC 25586, and V. parvula DSM, the anaerobic F. nucleatum ATCC 25586 and V. parvula DSM 2008, and the total mixed bacteria from pooled saliva and supra- and subgingival plaques of volunteers were all treated and compared. aPDT-VIS-wIRA with LE as PS significantly (p < 0.008) reduced the growth of all tested Gram-positive, Gram-negative, as well as aerobic and anaerobic bacterial strains, whereas without irradiation no reductions were seen (p < 0.0001). NaCl, with or without irradiation, was ineffective. After treatment with CHX 0.2%, the highest killing rate (100%) was observed, and no bacteria (0 log10 CFU) were cultivable. The method also significantly reduced all of the bacteria present in saliva and in the gingival biofilms. Three-dimensional visualization of viable and non-viable microorganisms revealed that LE penetrated deeper into the cell wall layers than CHX 0.2%. LE was an appropriate PS for eradicating microorganisms with VIS-wIRA, either in their planktonic form or in saliva and gingival plaque biofilms. These results encourage further investigation in order to determine which LE compounds contribute to the photosensitizing effect and to evaluate the size of the effect on maintaining oral health.


Myricetin and morin hydrate inhibit amyloid fibril formation of bovine α-lactalbumin (BLA).
Amyloid fibrils are self-assembled aggregates of proteins and peptides that can lead to a broad range of diseases called amyloidosis. So far, no definitive and approved treatment to target directly amyloid fibrils has been introduced. Nevertheless, the search for small molecules with ability to inhibit and suppress fibril formation is an active and promising area of the research. Herein, the binding interactions and inhibitory effects of myricetin and morin hydrate on the in vitro fibrillation of bovine α-lactalbumin (BLA) have been investigated. The intrinsic fluorescence of BLA was quenched by myricetin and morin hydrate through combination of the static and dynamic quenching along with non-radiative Förster energy transfer mechanisms. The binding of these two flavonoids to BLA were not accompanied by major alteration in the conformation of BLA as evidenced by CD studies. The results of the fluorescence quenching analyses indicated almost the same binding affinities of myricetin and morin hydrate toward BLA (Kb ~ 106 M-1). However, the results of thioflavin T (ThT) assays showed that myricetin is a stronger inhibitor against BLA fibrillation compared to morin hydrate.


Efficacy and safety of eptastigmine for the treatment of patients with Alzheimer's disease.
To evaluate the efficacy and safety of eptastigmine in patients with moderate to moderately severe AD. Eptastigmine is a centrally acting cholinesterase inhibitor. The study was carried out according a multicenter, randomized, double-blinded, placebo-controlled, parallel-group design. Patients received a 24-week treatment with placebo or eptastigmine 15 mg or 20 mg three times daily after a 4-week, stepwise dose escalation. The effects of treatment on cognition, global function, and activities of daily living were evaluated with the Alzheimer's Disease Assessment Cognitive Subscale (ADAS-Cog), the Clinician's Interview-Based Impression of Change Plus (CIBIC-Plus), and the Instrumental Activities of Daily Living scale (IADL), respectively. Thirty-six centers recruited 491 patients: 164 on placebo, 166 on eptastigmine 15 mg three times daily, and 161 on eptastigmine 20 mg three times daily. Percentages of patients completing double-blinded treatment were 87% in the placebo group and 86% in both the eptastigmine-treated groups. At the end of treatment, the intent-to-treat analysis on 463 patients showed a dose-dependent effect of eptastigmine on all efficacy variables, with a statistically significant effect of the 20 mg three times daily dose compared with placebo on the ADAS-Cog, CIBIC-Plus, and IADL. Patients on eptastigmine 15 mg three times daily performed significantly better than placebo-treated patients only on the ADAS-Cog. Eleven patients on placebo (7%), 13 patients on eptastigmine 15 mg three times daily (8%), and 12 patients on eptastigmine 20 mg three times daily (8%) discontinued study treatment because of adverse events. Adverse events were recorded in 49% of patients on placebo compared with 54% on eptastigmine 15 mg three times daily and 48% on eptastigmine 20 mg three times daily. Cholinergic side effects (nausea, vomiting, diarrhea, and abdominal pain) were reported with similar frequency in the eptastigmine- and placebo-treated patients. There was a dose-dependent transient and mild neutropenic effect associated with eptastigmine treatment, and one patient on 20 mg three times daily had an asymptomatic pancytopenia. Eptastigmine produces significant cognitive, clinical, and functional benefits in patients with probable AD. Although the cholinergic tolerability of eptastigmine was found to be favorable, its potential adverse hematologic effects limit its clinical utility.


Mobile telephony and its effects on human health
In recent years, there has been a rapid increase in the number of electromagnetic radiation sources such as mobile phones and base stations of mobile telephony. This radiation has been classified by the International Agency for Research on Cancer as a possible human carcinogen (group 2B). For this reason, many studies have been carried out on the impact of mobile telephony on human health. The largest of the experiments were carried out on animals. Due to the divergent results of many studies, there was no clear answer on the possible carcinogenic effects of this type of radiation on health. Detection of cancer shortly after an exposure is the most difficult task in analyzing the results. Some of the studies require repetition and verification of the results. In the case of negative effects of electromagnetic fields on brain activity, sleep, heart rate, cognitive function and blood pressure, no consistent evidence has been obtained either. In view of the increasing popularity of mobile phones, their location at short distances from the body (mainly the head) and the development of mobile phone technologies (which entails an increase in the number of base stations), further research, especially among young people, is needed. The duration of human exposure to electromagnetic radiation is subject to an increase and only further research can provide an answer on the possible negative effects of mobile phones and base stations.


Protection against a mixed SHIV challenge by a broadly neutralizing antibody cocktail.
HIV-1 sequence diversity presents a major challenge for the clinical development of broadly neutralizing antibodies (bNAbs) for both therapy and prevention. Sequence variation in critical bNAb epitopes has been observed in most HIV-1-infected individuals and can lead to viral escape after bNAb monotherapy in humans. We show that viral sequence diversity can limit both the therapeutic and prophylactic efficacy of bNAbs in rhesus monkeys. We first demonstrate that monotherapy with the V3 glycan-dependent antibody 10-1074, but not PGT121, results in rapid selection of preexisting viral variants containing N332/S334 escape mutations and loss of therapeutic efficacy in simian-HIV (SHIV)-SF162P3-infected rhesus monkeys. We then show that the V3 glycan-dependent antibody PGT121 alone and the V2 glycan-dependent antibody PGDM1400 alone both fail to protect against a mixed challenge with SHIV-SF162P3 and SHIV-325c. In contrast, the combination of both bNAbs provides 100% protection against this mixed SHIV challenge. These data reveal that single bNAbs efficiently select resistant viruses from a diverse challenge swarm to establish infection, demonstrating the importance of bNAb cocktails for HIV-1 prevention.


Dynamics of hypervariable region 1 variation in hepatitis C virus infection and correlation with clinical and virological features of liver disease.
Hepatitis C virus (HCV) infection is a dynamic process during which molecular variants are continuously selected as the result of virus adaptation to the host. Understanding the nature of HCV genetic variation is central to current theories of pathogenesis and immune response. We prospectively studied hypervariable region 1 (HVR1) variation in the E2 gene of 36 hepatitis C patients, including 10 asymptomatic carriers, followed up for 1 to 2 years. Sequence changes in single and consecutive serum samples were assessed and correlated with clinical and virological parameters of liver disease. A region of the E1 gene was sequenced for comparison in 3 subjects. HVR1 heterogeneity at single time points widely varied in individual patients, did not increase cumulatively over the follow-up period, and did not correlate with HVR1 evolutionary rates. Conversely, the process of HVR1 sequence diversification, although differed considerably among patients, was stable over time and directly correlated with infections by HCV type 2, lower alanine aminotransferase (ALT) levels, and absence of cirrhosis. HCV carriers showed the highest HVR1 variation rates. Our findings indicate that HVR1 variation has an adaptive significance and is associated with favorable features of liver disease and suggest that prospective, rather than static, observations are required to model the process of HCV variation.


Screening and quantification of hypnotic sedatives in serum by capillary gas chromatography with a nitrogen-phosphorus detector, and confirmation by capillary gas chromatography-mass spectrometry.
We describe a quantitative screen for hypnotic-sedative drugs in which we use capillary gas chromatography with a nitrogen-phosphorus detector (GC/NPD) as the primary method and capillary gas chromatography-mass spectrometry (GC-MS) for confirmation. GC retention times of the acid-extracted underivatized drugs were stable (CVs less than 1%), and the detector response varied linearly over a 20-fold concentration range with a mean correlation coefficient for 11 drugs of 0.989. The limits of detection were satisfactory (0.5 mg/L in a 0.5-mL serum sample and 1-microL injection volume), as were precision (average CV 5.2% within day, 6.4% between day). The complementary use of capillary GC-MS not only unambiguously confirms presumptive peaks identified by GC, but also prevents reports of false positives and identifies compounds not included in the quantitative GC screen that may be listed in the GC-MS library.


Scanning electron microscope evidence for bacterial colonization of a drinking-water distribution system.
The surfaces of water distribution mains and suspended particulate matter from drinking water were examined by using scanning electron microscopy to investigate the nature and extent of association of microorganisms with these surfaces. In addition, X-ray energy-dispersive microanalysis was used to determine the elemental constitution of the pipe surface. Though distributed sparsely and randomly along the pipe surface, a variety of morphologically distinguishable bacteria-like structures and microcolonies were observed. The morphologies of the individual cells varied form chain-forming cocci to filamentous and prosthecate cell types. The iron-oxidizing bacterium Gallionella, recognized by its characteristic helical stalks, was observed both in water samples and attached to pipe surfaces. Attachment of some microbes to the pipe surface was apparently mediated by extracellular fibrillar appendages. Large numbers of rod-shaped bacteria were also evident adhering to the surfaces of suspended detritus or silt particles recovered from water samples by filtration. X-ray energy scans of the pipe surface revealed the presence of five major elemental constituents including silicon, phosphorous, sulfur, calcium, and iron. Smaller quantities of the elements zinc, magnesium, aluminum, potassium, and manganese were also detected. The public health significance of sessile microbial communities in drinking-water distribution systems is discussed.


Association between lipoprotein lipase (LPL) gene and blood lipids: a common variant for a common trait?
S447X, a serine substitution by a stop codon on base 99 of exon 9 of the lipoprotein lipase (LPL) gene, has beneficial effects on blood lipids. Other LPL alleles are associated with lipid levels, but whether one of these variants predominates remains elusive. We performed a systematic survey to identify single-nucleotide polymorphisms (SNPs) in all 10 LPL exons and flanking regions by resequencing the gene in 95 subjects. Of 24 variants, 14 were common (> or = 3%). We assayed the common SNPs in 186 cases with atherogenic lipid profiles (low HDL, high LDL) and 185 nonatherogenic controls (high HDL, low LDL). Only S447X and exons 6 (base +73) and 10 (base -11) were individually associated with case-control status (P<0.05, adjusted for major nongenetic covariates with known lipid effects). There were no significant SNP x gender interactions. In adjusted multi-SNP and haplotypic analyses, S447X was interpretable as the sole predictor, with a 2-3-fold reduction in the odds of being atherogenic vs. nonatherogenic (adjusted OR, 0.39; 95% CI, 0.21-0.73). S447X and base -11 of exon 10 were statistically interchangeable because they are strongly associated (r=0.92, P<0.0001), but we posit that the LPL association with lipid profile is more likely attributable to the functional S447X rather than the nonfunctional exon 10 SNP. It appears that the S447X variant of LPL may be another rare example (like APOE4, factor V-Leiden, and PPAR gamma Pro12Ala) of a common variant predisposing to a common disorder.


Efficacy and safety of autologous platelet-rich plasma for diabetic foot ulcers: a systematic review and meta-analysis.
To investigate the efficacy and safety of autologous platelet-rich plasma (au-PRP) for diabetic foot ulcer (DFU) treatment. We conducted database searches (MEDLINE, EMBASE, evidence-based medicine reviews: CENTRAL, PubMed, and Web of Science) and reference mining for randomised controlled trials from inception to 23 January 2022. Results were scrutinised, data were extracted and research quality was investigated by two independent authors. Primary outcome was the proportion of complete ulcer healing. Secondary outcomes included both the mean time to complete healing and the incidence of adverse events. Statistical analyses were performed in RevMan 5.4 (Cochrane, UK). Kaplan-Meier curves for time to complete healing were pooled in R software (version 4.1.2) (R Foundation, Austria). Of the 231 records identified, 17 studies with a total of 1303 participants (649 randomised to the au-PRP group and 654 to a standard of care (SOC) group) met the eligibility criteria and were included in our study. Compared with SOC, au-PRP appeared to promote the complete healing rate (odds ratio (OR): 2.11; 95% Confidence Interval: 1.55-2.86). Au-PRP also appeared to significantly shorten complete healing time (mean duration: -19.04 days; 95%CI: -20.46--17.61]). There was no significant difference on adverse events. Results were robust on sensitivity analyses. Based on the findings of this review and meta-analysis, Au-PRP is an effective and safe adjuvant therapy for DFUs.


Factors Affecting Menstrual Hygiene Management Practice Among School Adolescents in Ambo, Western Ethiopia, 2018: A Cross-Sectional Mixed-Method Study.
Menstruation is still regarded as something unclean in many parts of Ethiopia. Despite the challenges related to menstrual hygiene management, it has been routinely overlooked in the school setting. The existing evidence on menstrual hygiene management lacks objective measurement approaches. Therefore, the aim of this study was to assess factors affecting menstrual hygiene management practice among school adolescents in Ambo, Western Ethiopia. An institutional-based mixed-method cross-sectional study was conducted from 1 to 30 March 2018 with 364 high school females. A systematic random sampling and purposive sampling technique were used to select study participants. We used a self-administered questionnaire, observational checklists, and focus group discussion. Data were analyzed using SPSS statistical software and thematic analysis. Multiple logistic regression analyses were done. With 95% CI, P< 0.05 was considered the level of significance. The prevalence of unsafe menstrual hygiene management practice was 53.6%, and it was affected by factors including age of the females [AOR: 0.16 (0.045, 0.57), P=0.005], frequency of discussing menses [AOR: 0.30, 95% CI: 0.13, 0.71, P-value = 0.006], and source of information about menses [AOR: 3.75, 95% CI: 1.75, 8.00) P= 0.001]. Lack of information about how to manage menses safely and lack of sufficient WASH facility were also identified. High prevalence of unsafe menstrual hygiene management in the study area implies that urgent measures are needed from the concerned body. The school environments were not female friendly for managing their menses safely highlighting that the water, sanitation, and hygiene facilities in the schools need urgent solutions.


Adult Onset Still's Disease Presenting with Acute Respiratory Distress Syndrome: Case Report and Review of the Literature.
Adult-onset Still's disease (AOSD) is a systemic inflammatory disorder characterized by rash, leukocytosis, fevers, and arthralgias. Pulmonary involvement has been reported rarely in AOSD, but acute respiratory distress syndrome (ARDS) is extremely rare and potentially fatal and must be recognized as potential manifestation of underlying AOSD. We present a case of AOSD manifested by ARDS and review the previously reported cases in Medline/Pub med. Including this case, 19 cases of AOSD complicated with ARDS have been reported in the literature. It is important to recognize ARDS as a manifestation of AOSD so that proper diagnostic and therapeutic management can be initiated promptly.


Intracranial aneurysms in the infant population: an institutional case series and individual participant data meta-analysis.
Infantile intracranial aneurysms are exceedingly rare. The goal of this study was to evaluate an institutional case series of infantile intracranial aneurysms, as well as those reported in the contemporary literature, to determine their demographics, presentation, management, and long-term outcome. A comprehensive literature review from 1980 to 2020 was performed to identify individual cases of intracranial aneurysms in the infantile population ≤ 2 years of age. Additional cases from the authors' institution were identified during the same time period. An individual participant data meta-analysis (IPDMA) was performed, abiding by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Patient demographic, radiographic, and clinical information was obtained. Descriptive statistical data were recorded, and multivariate logistic regression analyses were performed. Patient data were obtained for 133 patients from 87 articles in the literature. Ten additional patients at the authors' institution were also identified, for a total of 143 patients included in the IPDMA. The majority (72.7%) of this cohort consisted of idiopathic aneurysms, while 13.3% were posttraumatic pseudoaneurysms, 9.8% were infectious mycotic aneurysms, and 4.2% were aneurysms associated with a systemic connective tissue disorder or vasculitis. The mean age at presentation was 6.6 months. The majority of infants (97.9%) harbored only 1 aneurysm, and hemorrhage was the most common presenting feature (78.3%). The mean aneurysm size was 14.4 mm, and giant aneurysms ≥ 25 mm comprised 12.9% of the cohort. Most aneurysms occurred in the anterior circulation (80.9%), with the middle cerebral artery (MCA) being the most commonly affected vessel (51.8%). Management strategies included open surgical aneurysm ligation (54.0%), endovascular treatment (35.0%), surgical decompression without aneurysm treatment (4.4%), and medical supportive management only (13.9%). Surgical aneurysm ligation was more commonly performed for MCA and anterior cerebral artery aneurysms (p = 0.004 and p = 0.015, respectively), while endovascular techniques were favored for basilar artery aneurysms (p = 0.042). The mean follow-up period was 29.9 months; 12.4% of the cohort died, and 67.0% had a favorable outcome (Glasgow Outcome Scale score of 5). This study is, to the authors' knowledge, the largest analysis of infantile intracranial aneurysms to date. The majority were idiopathic aneurysms involving the anterior circulation. Surgical and endovascular techniques yielded equally favorable outcomes in this cohort. Long-term outcomes in the infantile population compared favorably to outcomes in adults.


Role of CD44 in the reaction of vascular smooth muscle cells to arterial wall injury.
CD44, the principal receptor for hyaluronic acid, is a widely distributed cell surface proteoglycan involved in cellular activation, proliferation, and migration. These processes are also central to the vascular smooth muscle cell's response to arterial wall injury. We evaluated the expression of CD44 and its isoform, CD44-V6, on vascular smooth muscle cells in vitro and in vivo and assessed the role of CD44 in DNA synthesis. Cultured vascular smooth muscle cells expressed CD44 and CD44-V6 at levels equal to or higher than those of the beta 1 and beta 2 integrins. In a rat carotid artery balloon injury model, CD44 and CD44-V6 mRNAs were unregulated in vascular smooth muscle cells after injury, and CD44 protein expression was greatest at the luminal edge of the growing neointima. CD44-expressing smooth muscle cells proliferated actively, and hyaluronic acid expression increased after injury in a temporal pattern similar to that of CD44. Through binding to hyaluronic acid, CD44 augmented DNA synthesis in cultured human and rat smooth muscle cells by 48 +/- 7.8 and 100 +/- 12.5%, respectively, an effect inhibited by an anti-CD44 antibody that blocked hyaluronate binding. These observations support a role for CD44 in the reaction of vascular smooth muscle cells to arterial wall injury.


Transglutaminase-2 mediates calcium-regulated crosslinking of the Y-box 1 (YB-1) translation-regulatory protein in TGFβ1-activated myofibroblasts.
Myofibroblast differentiation is required for wound healing and accompanied by activation of smooth muscle α-actin (SMαA) gene expression. The stress-response protein, Y-box binding protein-1 (YB-1) binds SMαA mRNA and regulates its translational activity. Activation of SMαA gene expression in human pulmonary myofibroblasts by TGFβ1 was associated with formation of denaturation-resistant YB-1 oligomers with selective affinity for a known translation-silencer sequence in SMαA mRNA. We have determined that YB-1 is a substrate for the protein-crosslinking enzyme transglutaminase 2 (TG2) that catalyzes calcium-dependent formation of covalent γ-glutamyl-isopeptide linkages in response to reactive oxygen signaling. TG2 transamidation reactions using intact cells, cell lysates, and recombinant YB-1 revealed covalent crosslinking of the 50 kDa YB-1 polypeptide into protein oligomers that were distributed during SDS-PAGE over a 75-250 kDa size range. In vitro YB-1 transamidation required nanomolar levels of calcium and was enhanced by the presence of SMαA mRNA. In human pulmonary fibroblasts, YB-1 crosslinking was inhibited by (a) anti-oxidant cystamine, (b) the reactive-oxygen antagonist, diphenyleneiodonium, (c) competitive inhibition of TG2 transamidation using the aminyl-surrogate substrate, monodansylcadaverine, and (d) transfection with small-interfering RNA specific for human TG2 mRNA. YB-1 crosslinking was partially reversible as a function of oligomer-substrate availability and TG2 enzyme concentration. Intracellular calcium accumulation and peroxidative stress in injury-activated myofibroblasts may govern SMαA mRNA translational activity during wound healing via TG2-mediated crosslinking of the YB-1 mRNA-binding protein.


The C-terminal GGAP motif of Hsp70 mediates substrate recognition and stress response in yeast.
The allosteric coupling of the highly conserved nucleotide- and substrate-binding domains of Hsp70 has been studied intensively. In contrast, the role of the disordered, highly variable C-terminal region of Hsp70 remains unclear. In many eukaryotic Hsp70s, the extreme C-terminal EEVD motif binds to the tetratricopeptide-repeat domains of Hsp70 co-chaperones. Here, we discovered that the TVEEVD sequence of Saccharomyces cerevisiae cytoplasmic Hsp70 (Ssa1) functions as a SUMO-interacting motif. A second C-terminal motif of ∼15 amino acids between the α-helical lid and the extreme C terminus, previously identified in bacterial and eukaryotic organellar Hsp70s, is known to enhance chaperone function by transiently interacting with folding clients. Using structural analysis, interaction studies, fibril formation assays, and in vivo functional assays, we investigated the individual contributions of the α-helical bundle and the C-terminal disordered region of Ssa1 in the inhibition of fibril formation of the prion protein Ure2. Our results revealed that although the α-helical bundle of the Ssa1 substrate-binding domain (SBDα) does not directly bind to Ure2, the SBDα enhances the ability of Hsp70 to inhibit fibril formation. We found that a 20-residue C-terminal motif in Ssa1, containing GGAP and GGAP-like tetrapeptide repeats, can directly bind to Ure2, the Hsp40 co-chaperone Ydj1, and α-synuclein, but not to the SUMO-like protein SMT3 or BSA. Deletion or substitution of the Ssa1 GGAP motif impaired yeast cell tolerance to temperature and cell-wall damage stress. This study highlights that the C-terminal GGAP motif of Hsp70 is important for substrate recognition and mediation of the heat shock response.


[Changes in the contractile myocardium in acute rhythm disorders].
Experiments in rabbits were carried out to study the morphology of contractile myocardium in arrhythmias caused by electric stimulation of previously intact myocardium (1st group) and in those caused by inhalation of ammonia steam against the background of exogenous poisoning with barium chloride (2nd group). Morphological lesions of contractile myocardium consisting in granular an vacuolar dystrophy at 1--3 days of the experiment and focal myocytolysis at 7 days were demonstrated. In animals of the 2nd group, morphological lesions in the myocardium were more severe and by the 7th day of the experiment included both focal myocytolysis and contracture alterations. Changes in the blood vessels of the microcirculation bed were similar in both groups consisting of vascular spasm followed by paresis, plethora, diapedetic hemorrhages and early signs of thrombus formation. The latter was observed earlier in animals of the 2nd group. The observed morphological changes in contractile myocardium may be regarded as one of the forms on non-coronarogenic myocardiopathies.


[Fever in patients of a Service of Internal Medicine: prospective study in 204 cases].
Fever in hospitalized patients (FHP) is a difficult problem. We study clinical characteristics and predictive parameters of infection, sepsis and outcome. Prospective study of 204 patients with fever > 38 degrees C admitted in an Internal Medicine ward. In each patient clinical evaluation, complete blood count (CBC) urinalysis (UA), C-reactive protein (CRP), chest X- ray (CXR), blood and urine cultures were performed. 115 patients suffered infection (75.9% nosocomial, 7.4% sepsis), 35 had a non-infectious etiology and 54 an unknown cause. CBC and CRP did not distinguish infection, bacteremia or sepsis. In UA, positive nitrites, leukocytes and bacteriuria were predictive of infection. 18.6% of the patients were diagnosed by CXR. 18% of urine cultures and 13% of blood cultures were positive. 71.6% received antibiotics (deemed unnecessary in 18%). Microbiological results modified 25% of initial treatments. Average hospital stay was 17 days; 16.7% died. Chronic lung disease, fever duration, dysphagia and urinary tract alterations predict infection. Obesity, diabetes mellitus, liver failure, immunocompromised host, central vascular access, fever duration and nosocomial infection predict bacteremia. Sepsis is more prevalent in males, with malignancy and vascular or nosocomial infection. Chronic liver disease, nosocomial infection, severe lung infection and sepsis increase mortality. Clinical diagnosis of FHP is inaccurate. Infection is it s most frequent cause; UA, CXR, and blood and urine cultures are useful. FHP increases mortality and hospital stay. We have established predictable models of infection, bacteremia, sepsis and mortality. However, its sensibility and specificity are low.


Somatic cell count assessment at the quarter or cow milking level.
The aim was to investigate whether on-line somatic cell count (SCC) assessment, when combined with electrical conductivity (EC), should be implemented at the udder quarter or at the cow level. Data were collected from 3 farms with automatic milking systems, resulting in 3,191 quarter milkings used in the analyses. Visual observations of foremilk and quarter milk samples for laboratory SCC analysis were used to define 2 gold standards. One was based on visual observation only and the other was based on a combination of visual observation and SCC (using a reference value of 500,000 cells/mL), which means that a quarter milking must have visually abnormal milk as well as an increased SCC to be categorized positive. On-line SCC assessment took place at the quarter level during the first part of the milking. Composite cow level samples were used for laboratory SCC analysis and to compare the performance of SCC assessment at quarter and cow levels. The EC at the quarter level was measured by in-line sensors of the automatic milking system. Alerts for SCC indicators were calculated based on straightforward reference values. Alerts for EC were based on straightforward reference values, or on interquarter ratios. The latter was calculated by dividing the value of a given quarter by the average value of the 2 lowest quarters of that milking. The EC and SCC indicators were combined with either a Boolean "and" or "or" function. Receiver operating characteristic curves were used to visually present results using different threshold values. Sensitivity, specificity, and success rate at the quarter level and false alert rate per 1,000 cow milkings were used to compare indicators at given sensitivity or specificity levels. Quarter level SCC assessment was superior to cow level assessment (transformed partial area under the curve=0.70 vs. 0.62) when combined with EC measurement at quarter level. When aiming for the same sensitivity level (e.g., 50%) with all visual abnormal milk as the gold standard, more false alerts were generated with cow level assessment (137 per 1,000 cow milkings) compared with quarter level SCC assessment (75 per 1,000 cow milkings). As a comparison, using EC alone resulted in 292 false alerts per 1,000 cow milkings in the same situation. Therefore, it is concluded that quarter level SCC assessment was superior to cow level assessment when combined with EC measurement at quarter level.


General Anaesthesia Protocols for Patients Undergoing Electroconvulsive Therapy: Retrospective analysis of 504 sessions over a five-year period at a tertiary care hospital in Oman.
This study aimed to review general anaesthesia protocols for patients undergoing electroconvulsive therapy (ECT) at a tertiary care hospital in Oman, particularly with regards to clinical profile, potential drug interactions and patient outcomes. This retrospective study took place at the Sultan Qaboos University Hospital (SQUH), Muscat, Oman. The electronic medical records of patients undergoing ECT at SQUH between January 2010 and December 2014 were reviewed for demographic characteristics and therapy details. A total of 504 modified ECT sessions were performed on 57 patients during the study period. All of the patients underwent a uniform general anaesthetic regimen consisting of propofol and succinylcholine; however, they received different doses between sessions, as determined by the treating anaesthesiologist. Variations in drug doses between sessions in the same patient could not be attributed to any particular factor. Self-limiting tachycardia and hypertension were periprocedural complications noted among all patients. One patient developed aspiration pneumonitis (1.8%). All patients undergoing ECT received a general anaesthetic regimen including propofol and succinylcholine. However, the interplay of anaesthetic drugs with ECT efficacy could not be established due to a lack of comprehensive data, particularly with respect to seizure duration. In addition, the impact of concurrent antipsychotic therapy on anaesthetic dose and subsequent complications could not be determined.


Feasibility of repairing glomerular basement membrane defects in Alport syndrome.
Alport syndrome is a hereditary glomerular disease that leads to kidney failure. It is caused by mutations affecting one of three chains of the collagen α3α4α5(IV) heterotrimer, which forms the major collagen IV network of the glomerular basement membrane (GBM). In the absence of the α3α4α5(IV) network, the α1α1α2(IV) network substitutes, but it is insufficient to maintain normal kidney function. Inhibition of angiotensin-converting enzyme slows progression to kidney failure in patients with Alport syndrome but is not a cure. Restoration of the normal collagen α3α4α5(IV) network in the GBM, by either cell- or gene-based therapy, is an attractive and logical approach toward a cure, but whether or not the abnormal GBM can be repaired once it has formed and is functioning is unknown. Using a mouse model of Alport syndrome and an inducible transgene system, we found that secretion of α3α4α5(IV) heterotrimers by podocytes into a preformed, abnormal, filtering Alport GBM is effective at restoring the missing collagen IV network, slowing kidney disease progression, and extending life span. This proof-of-principle study demonstrates the plasticity of the mature GBM and validates the pursuit of therapeutic approaches aimed at normalizing the GBM to prolong kidney function.


Quantification of False Vocal Fold Hyperfunction During Quiet Breathing in Muscle Tension Dysphonia.
False vocal fold (FVF) hyperfunction during phonation is thought to be a diagnostic sign of primary muscle tension dysphonia (pMTD). However, hyperfunctional patterns with phonation are also observed in typical speakers. This study tested the hypothesis that FVF posturing during quiet breathing, as measured by the curvature of FVF, could differentiate patients with pMTD from typical speakers. Laryngoscopic images were collected prospectively in 30 subjects with pMTD and 33 typical speakers. Images were acquired at the end of expiration and maximal inspiration during quiet breathing, during sustained /i/, and during loud phonation before and after a 30-min vocal loading task. The FVF curvature (degree of concavity/convexity) was quantified using a novel curvature index (CI, >0 for hyperfunctional/convex, <0 for "relaxed"/concave) and compared between the two groups. At end-expiration, the pMTD group adopted a convex FVF contour, whereas the control group adopted a concave FVF contour (mean CI 0.123 [SEM 0.046] vs. -0.093 [SEM 0.030], p = 0.0002) before vocal loading. At maximal inspiration, the pMTD group had a neutral/straight FVF contour, whereas the control group had a concave FVF contour (mean CI 0.012 [SEM 0.038] vs. -0.155 [SEM 0.018], p = 0.0002). There were no statistically significant differences in FVF curvature between groups in either the sustained voiced or loud conditions. Vocal loading did not change any of these relationships. A hyperfunctional posture of the FVFs during quiet breathing especially at end-expiration may be more indicative of a hyperfunctional voice disorder than supraglottic constriction during voicing. 3 Laryngoscope, 133:3449-3454, 2023.


Anaphylactic Shock to Intravenous Indocyanine Green During a Robotic Right Colectomy.
Intravenous indocyanine green (IV ICG) is regarded as a safe immunofluorescence agent used to assess bowel perfusion prior to creating bowel anastomoses and aids in ureter identification during intra-abdominal surgery. We report the first instance of anaphylactic shock to IV ICG after prior toleration of ICG via an intra-ureteral route. Shortly after administering IV ICG, our patient became hypotensive and hypoxic requiring chest compressions, vasoactive medications, and thoracostomy tubes prior to identifying the symptoms as an allergic reaction. Anaphylaxis is not a recognized side effect of ICG and was not immediately considered. As ICG becomes increasingly utilized as an immunofluorescence agent among surgical specialties, increased awareness and recognition of anaphylactic shock as a potential side effect of ICG may lead to expedited diagnoses, treatment, and more critical evaluation of indications for future use. Additionally, our patient first tolerated intra-ureteral administration without a systemic reaction, suggesting a possible sensitization mechanism.


Protected carotid stenting in high-surgical-risk patients: the ARCHeR results.
Carotid endarterectomy is the standard of care for most patients with severe extracranial carotid bifurcation disease. However, its safety and efficacy in patients with significant surgical risk are unclear. The ARCHeR (ACCULINK for Revascularization of Carotids in High-Risk patients) trial was performed to determine whether carotid artery stenting with embolic protection is a safe and effective alternative to endarterectomy in high-surgical-risk patients. The ARCHeR trial is a series of three sequential, multicenter, nonrandomized, prospective studies. Forty-eight sites enrolled 581 high-surgical-risk patients between May 2000 and September 2003. Patients with severe carotid artery stenosis (angiographically defined, symptomatic > or =50%, or asymptomatic > or =80%) had an ACCULINK nitinol stent implanted. The ACCUNET filter embolic protection system was added to the procedure in the final 2 studies (422 patients). The primary efficacy end point was a composite of periprocedural (< or =30 days) death, stroke, and myocardial infarction, plus ipsilateral stroke between days 31 and 365. The 30-day rate of death/stroke/myocardial infarction was 8.3% (95% confidence interval [CI], 6.2%-10.8%), and that of stroke/death was 6.9% (95% CI, 5.0%-9.3%). Most (23/32) strokes were minor, of which more than half (12/23) returned to baseline National Institutes of Health Stroke Scale scores within 30 days. The 30-day major/fatal stroke rate was 1.5% (95% CI, 0.7%-2.9%). No hemorrhagic strokes were observed in the study. Ipsilateral cerebrovascular accident occurred in 1.3% between 30 days and 1 year, thus giving a primary composite end point of 30-day death/stroke/myocardial infarction plus ipsilateral stroke at 1 year of 9.6% (95% CI, 7.2%-12.0%), which is below the 14.4% historical control comparator. Target lesion revascularization at 12 months and 2 years was 2.2% and 2.9%, respectively. The ARCHeR results demonstrate that extracranial carotid artery stenting with embolic filter protection is not inferior to historical results of endarterectomy and suggest that carotid artery stenting is a safe, durable, and effective alternative in high-surgical-risk patients.


Modulators of Enterococcus faecalis Cell Envelope Integrity and Antimicrobial Resistance Influence Stable Colonization of the Mammalian Gastrointestinal Tract.
The Gram-positive bacterium Enterococcus faecalis is both a colonizer of the gastrointestinal tract (GIT) and an agent of serious nosocomial infections. Although it is typically required for pathogenesis, GIT colonization by E. faecalis is poorly understood. E. faecalis tolerates high concentrations of GIT antimicrobials, like cholate and lysozyme, leading us to hypothesize that resistance to intestinal antimicrobials is essential for long-term GIT colonization. Analyses of E. faecalis mutants exhibiting defects in antimicrobial resistance revealed that IreK, a determinant of envelope integrity and antimicrobial resistance, is required for long-term GIT colonization. IreK is a member of the PASTA kinase protein family, bacterial transmembrane signaling proteins implicated in the regulation of cell wall homeostasis. Among several determinants of cholate and lysozyme resistance in E. faecalis, IreK was the only one found to be required for intestinal colonization, emphasizing the importance of this protein to enterococcal adaptation to the GIT. By studying ΔireK suppressor mutants that recovered the ability to colonize the GIT, we identified two conserved enterococcal proteins (OG1RF_11271 and OG1RF_11272) that function antagonistically to IreK and interfere with cell envelope integrity, antimicrobial resistance, and GIT colonization. Our data suggest that IreK, through its kinase activity, inhibits the actions of these proteins. IreK, OG1RF_11271, and OG1RF_11272 are found in all enterococci, suggesting that their effect on GIT colonization is universal across enterococci. Thus, we have defined conserved genes in the enterococcal core genome that influence GIT colonization through their effect on enterococcal envelope integrity and antimicrobial resistance.


Forskolin and phorbol esters reduce the same potassium conductance of mouse neurons in culture.
Second messenger systems may modulate neuronal activity through protein phosphorylation. However, interactions between two major second messenger pathways, the cyclic AMP and phosphatidylinositol systems, are not well understood. The effects of activators of cyclic AMP-dependent protein kinase and protein kinase C on resting membrane properties, action potentials, and currents recorded from mouse dorsal root ganglion neurons and cerebral hemisphere neurons grown in primary dissociated cell culture were investigated. Neither forskolin (FOR) nor phorbol 12,13-dibutyrate (PDBu) altered resting membrane properties but both increased the duration of calcium-dependent action potentials in both central and peripheral neurons. By means of the single-electrode voltage clamp technique, FOR and PDBu were shown to decrease the same voltage-dependent potassium conductance. This suggests that two independent second messenger systems may affect the same potassium conductance.


Meta-analysis of direct and indirect comparison of ticagrelor and prasugrel effects on platelet reactivity.
Studies have linked on-treatment platelet reactivity (PR) to adverse clinical outcomes. Because new P2Y12 inhibitors (prasugrel and ticagrelor) have been predominantly tested against clopidogrel, data on pharmacodynamic comparisons between these 2 drugs are scarce. We compared ticagrelor with prasugrel in a network meta-analysis. PubMed, Cochrane, and EMBASE were searched for studies assessing PR in patients with coronary artery disease treated with ticagrelor or prasugrel. All studies using prasugrel and/or ticagrelor providing platelet function measurement data using VerifyNow P2Y12 reaction units (PRUs), platelet reactivity index (PRI) vasodilator-stimulated phosphoprotein phosphorylation, or maximal platelet aggregation (MPA) by light transmission aggregometry were considered eligible. Mixed treatment comparison models directly compared ticagrelor and prasugrel and indirectly compared them using clopidogrel as a comparator with data presented as mean difference (95% confidence interval). Data were extracted from 29 studies, including 5,395 patients. Compared with clopidogrel 75 mg, both prasugrel 10 mg and ticagrelor 90 mg twice daily were associated with lower PRU (mean difference -117 [-134.1, -100.5] and -159.7 [-182.6, -136.6], respectively), a lower PRI (-24.2 [-28.2, -20.3] and -33.6 [-39.9, -27.6], respectively), and lower MPA (-11.8 [-17, -6.3] and -20.7 [-28.5, -12.8], respectively). Similar results were obtained with clopidogrel 150 mg. Ticagrelor 90 mg twice daily was associated with lower PRU (-42.5 [-62.9, -21.9]), lower PRI (-9.3 [-15.6, -3.5]), and lower MPA (-8.9 [-16.4, -1.2]) compared with prasugrel 10 mg. In conclusion, our meta-analysis suggests that ticagrelor achieved significantly lower on-treatment PR compared with prasugrel, with both being superior to clopidogrel standard or high dose.


Effect of a center-based early childhood care and education program on child nutritional status: A secondary analysis of a stepped wedge cluster randomized controlled trial in rural Sindh, Pakistan.
High-quality early childhood care and education (ECCE) programs can positively impact children's development. However, as an unintended consequence, ECCE attendance may also affect children's nutritional status. We evaluated the effect of a center-based ECCE intervention on child nutritional outcomes in rural Pakistan. This study utilized data from a stepped-wedge cluster randomized controlled trial of a center-based ECCE program that trained female youth to run high-quality preschools for children aged 3.5-5.5 years (LEAPS program) in rural Sindh, Pakistan. The program did not include any school meals. A total of 99 village clusters were randomized to receive the LEAPS intervention in three steps, and repeated cross-sectional surveys were conducted to assess the impact on children (4.5-5.5 years old) at four time points. Intention-to-treat analyses with multi-level mixed-effect models were used to estimate the effect of the intervention on child anthropometric outcomes. The analysis included 3,858 children with anthropometric data from four cross-sectional survey rounds. The LEAPS intervention was found to have a positive effect on child HAZ (mean difference: 0.13 z-scores; 95% confidence interval (CI): 0.02, 0.24). However, there was a negative effect on weight-based anthropometric indicators, -0.29 WHZ (95% CI: -0.42, -0.15), -0.13 BMIZ (95% CI: -0.23, -0.03), and -0.16 MUACZ (95% CI: -0.25, -0.05). An exploratory analysis suggested that the magnitude of the negative effect of LEAPS on WHZ, BMIZ, and WAZ was greater in the survey round during the COVID-19 lockdown. The LEAPS intervention positively affected child linear growth but had negative effects on multiple weight-based anthropometric measures. ECCE programs in low- and middle-income country settings should evaluate the integration of nutrition-specific interventions (e.g., school lunch, counseling on healthy diets) and infection control strategies to promote children's healthy growth and development. ClinicalTrials.gov, NCT03764436, https://clinicaltrials.gov/ct2/show/NCT03764436.


Percutaneous renal cryoablation in obese and morbidly obese patients.
To compare percutaneous renal cryoablation complications and outcomes in obese and morbidly obese vs nonobese patients. Three hundred eighty-nine percutaneous cryoablation procedures were performed in 367 patients for treatment of 421 renal masses at our institution between 2003 and 2012. Patients were categorized into 3 groups on the basis of body mass index (BMI): nonobese (BMI <30.0 kg/m(2)), obese (BMI 30.0-39.9 kg/m(2)), and morbidly obese (BMI ≥40.0 kg/m(2)). Each group was retrospectively analyzed for major complications (Clavien ≥grade 3) and oncologic outcomes. One hundred eighty-nine renal cryoablation procedures (48.6%) were performed on nonobese patients, 161 (41.4%) on obese patients, and 39 (10.0%) on morbidly obese patients. Eleven (5.8%) major complications occurred in nonobese patients, 15 (9.3%) in obese patients, and 3 (7.7%) in morbidly obese patients. As such, there was no significant difference in the rate of major complications in obese (P = .23) or morbidly obese (P = .67) compared with nonobese patients. There was 1 ablation-related death from complications of urosepsis. Thirteen local treatment failures were identified, including 5 technical failures and 8 local tumor recurrences during median imaging follow-up of 18 months (interquartile range: 8-36). Six local treatment failures (3.2%) occurred in nonobese patients, 5 (2.9%) in obese patients, and 2 (4.8%) in morbidly obese patients. Again, no significant difference was noted in local treatment failure rate between obese (P = .96) or morbidly obese (P = .57) compared with nonobese patients. Percutaneous renal cryoablation complication rates and short-term outcomes in obese and morbidly obese patients are similar to those in nonobese patients.


Progress in pursuit of therapeutic A2A antagonists: the adenosine A2A receptor selective antagonist KW6002: research and development toward a novel nondopaminergic therapy for Parkinson's disease.
Research and development of the adenosine A2A receptor selective antagonist KW6002 have focused on developing a novel nondopaminergic therapy for Parkinson's disease (PD). Salient pharmacologic features of KW6002 were investigated in several animal models of PD. In rodent and primate models, KW6002 provides symptomatic relief from parkinsonian motor deficits without provoking dyskinesia or exacerbating existing dyskinesias. The major target neurons of the A2A receptor antagonist were identified as GABAergic striatopallidal medium spiny neurons. A possible mechanism of A2A receptor antagonist action in PD has been proposed based on the involvement of striatal and pallidal presynaptic A2A receptors in the "dual" modulation of GABAergic synaptic transmission. Experiments with dopamine D2 receptor knockout mice showed that A2A receptors can function and anti-PD activities of A2A antagonists can occur independent of the dopaminergic system. Clinical studies of KW6002 in patients with advanced PD with L-dopa-related motor complications yielded promising results with regard to motor symptom relief without motor side effects. The development of KW6002 represents the first time that a concept gleaned from A2A biologic research has been applied successfully to "proof of concept" clinical studies. The selective A2A antagonist should provide a novel nondopaminergic approach to PD therapy.


Chemical composition of Earth, Venus, and Mercury.
Model compositions of Earth, Venus, and Mercury are calculated from the premise that planets and chondrites underwent four identical fractionation processes in the solar nebula. Because elements of similar properties stay together in these processes, five constraints suffice to define the composition of a planet: mass of the core, abundance of U, and the ratios K/U, Tl/U, and FeO/(FeO + MgO). Complete abundance tables, and normative mineralogies, are given for all three planets. Review of available data shows only a few gross trends for the inner planets: FeO decreases with heliocentric distance, whereas volatiles are depleted and refractories are enriched in the smaller planets.


Bayesian generalized structured component analysis.
Generalized structured component analysis (GSCA) is a component-based approach to structural equation modelling, which adopts components of observed variables as proxies for latent variables and examines directional relationships among latent and observed variables. GSCA has been extended to deal with a wider range of data types, including discrete, multilevel or intensive longitudinal data, as well as to accommodate a greater variety of complex analyses such as latent moderation analysis, the capturing of cluster-level heterogeneity, and regularized analysis. To date, however, there has been no attempt to generalize the scope of GSCA into the Bayesian framework. In this paper, a novel extension of GSCA, called BGSCA, is proposed that estimates parameters within the Bayesian framework. BGSCA can be more attractive than the original GSCA for various reasons. For example, it can infer the probability distributions of random parameters, account for error variances in the measurement model, provide additional fit measures for model assessment and comparison from the Bayesian perspectives, and incorporate external information on parameters, which may be obtainable from past research, expert opinions, subjective beliefs or knowledge on the parameters. We utilize a Markov chain Monte Carlo method, the Gibbs sampler, to update the posterior distributions for the parameters of BGSCA. We conduct a simulation study to evaluate the performance of BGSCA. We also apply BGSCA to real data to demonstrate its empirical usefulness.


Use of initial risk assessment and recording as the main nursing intervention in identifying risk of falls.
The consequences of falls among hospital patients are a great problem, for the patient, the family and society, and cost billions of dollars. In Sweden, almost one-third of all hip fractures occur in the hospital population. Despite this, very few prevention strategies have been developed and tested. In this study, a risk assessment and recording programme in relation to the risk of falling among patients in a geriatric department at a Swedish hospital was implemented. The records of all patients admitted to a geriatric unit during one year, and a stratified random sample of patient records, constituting the control group from the year before, were reviewed. No recording of assessments regarding the patients' risk of falling, and no preventive nursing interventions, were found in the records of the control group. The study group, however, increased the recording of risk assessment to 96%. Only implemented nursing interventions were found in the patients' records, despite the fact that Swedish law makes it obligatory for the registered nurse to record both the planning and implementation of nursing care. In the study group there were explicit descriptions of problems of concern for nursing regarding the patients' risk of falling in less than one-third of the records, the nursing care plans were rare, and the evaluations were not satisfactory. Nursing interventions consisted mostly of information or education, promotion of patient participation, and structuring of the environment. There was no agreement on any standard-care plan. Recording of falls was found more often in the study group than in the control group (probably due to more careful recording), but the proportion of injuries in relation to falls was higher in the control group. The results of this study may be used as a baseline for developing a nursing strategy and documentation relating to falls.


Sire contribution to fertilization failure and early embryo survival in cattle.
Despite passing routine laboratory tests of semen quality, bulls used in artificial insemination (AI) exhibit a significant range in field fertility. The objective of this study was to determine whether subfertility in AI bulls is due to issues of sperm transport to the site of fertilization, fertilization failure, or failure of early embryo or conceptus development. In experiment 1, Holstein-Friesian bulls (3 high fertility, HF, and 3 low fertility, LF) were selected from the national population of AI bulls based on adjusted fertility scores from a minimum of 500 inseminations (HF: +4.37% and LF: -12.7%; mean = 0%). Superovulated beef heifers were blocked based on estimated number of follicles at the time of AI and inseminated with semen from HF or LF bulls (n = 3-4 heifers per bull; total 19 heifers). Following slaughter 7 d later, the number of corpora lutea was counted and the uteri were flushed. Recovered structures (oocytes/embryos) were classified according to developmental stage and stained with 4',6-diamidino-2-phenylindole to assess number of cells and accessory sperm. Overall recovery rate (total structures recovered/total corpora lutea) was 52.6% and was not different between groups. Mean (± standard error of the mean) number of embryos recovered per recipient was 8.7 ± 5.2 and 9.4 ± 5.5 for HF and LF, respectively. Overall fertilization rate of recovered structures was not different between groups. However, more embryos were at advanced stages of development (all blastocyst stages combined), reflected in a greater mean embryo cell number on d 7 for HF versus LF bulls. Number of accessory sperm was greater for embryos derived from HF than for LF bulls. The aim of experiment 2 was to evaluate the effect of sire fertility on survival of bovine embryos to d 15. Day 7 blastocysts were produced in vitro using semen from the same HF (n = 3) and LF (n = 3) bulls and transferred in groups of 5-10 to synchronized heifers (n = 7 heifers per bull; total 42 heifers). Conceptus recovery rate on d 15 was higher in HF (59.4%,) versus LF (45.0%). Mean length of recovered conceptuses for HF bulls was not affected by fertility status. In conclusion, while differences in field fertility among AI sires used in this study were not reflected in fertilization rate, differences in embryo quality were apparent as early as d 7. These differences likely contributed to the higher proportion of conceptuses surviving to d 15 in HF bulls.


Caspase-3 contributes to ZO-1 and Cl-5 tight-junction disruption in rapid anoxic neurovascular unit damage.
Tight-junction (TJ) protein degradation is a decisive step in hypoxic blood-brain barrier (BBB) breakdown in stroke. In this study we elucidated the impact of acute cerebral ischemia on TJ protein arrangement and the role of the apoptotic effector protease caspase-3 in this context. We used an in vitro model of the neurovascular unit and the guinea pig whole brain preparation to analyze with immunohistochemical methods the BBB properties and neurovascular integrity. In both methodological approaches we observed rapid TJ protein disruptions after 30 min of oxygen and glucose deprivation or middle cerebral artery occlusion, which were accompanied by strong caspase-3 activation in brain endothelial cells (BEC). Surprisingly only few DNA-fragmentations were detected with TUNEL stainings in BEC. Z-DEVD-fmk, an irreversible caspase-3 inhibitor, partly blocked TJ disruptions and was protective on trans-endothelial electrical resistance. Our data provide evidence that caspase-3 is rapidly activated during acute cerebral ischemia predominantly without triggering DNA-fragmentation in BEC. Further we detected fast TJ protein disruptions which could be partly blocked by caspase-3 inhibition with Z-DEVD-fmk. We suggest that the basis for clinically relevant BBB breakdown in form of TJ disruptions is initiated within minutes during ischemia and that caspase-3 contributes to this process.


A two-pronged structural analysis of retroviral maturation indicates that core formation proceeds by a disassembly-reassembly pathway rather than a displacive transition.
Retrovirus maturation involves sequential cleavages of the Gag polyprotein, initially arrayed in a spherical shell, leading to formation of capsids with polyhedral or conical morphology. Evidence suggests that capsids assemble de novo inside maturing virions from dissociated capsid (CA) protein, but the possibility persists of a displacive pathway in which the CA shell remains assembled but is remodeled. Inhibition of the final cleavage between CA and spacer peptide SP1/SP blocks the production of mature capsids. We investigated whether retention of SP might render CA assembly incompetent by testing the ability of Rous sarcoma virus (RSV) CA-SP to assemble in vitro into icosahedral capsids. Capsids were indeed assembled and were indistinguishable from those formed by CA alone, indicating that SP was disordered. We also used cryo-electron tomography to characterize HIV-1 particles produced in the presence of maturation inhibitor PF-46396 or with the cleavage-blocking CA5 mutation. Inhibitor-treated virions have a shell that resembles the CA layer of the immature Gag shell but is less complete. Some CA protein is generated but usually not enough for a mature core to assemble. We propose that inhibitors like PF-46396 bind to the Gag lattice where they deny the protease access to the CA-SP1 cleavage site and prevent the release of CA. CA5 particles, which exhibit no cleavage at the CA-SP1 site, have spheroidal shells with relatively thin walls. It appears that this lattice progresses displacively toward a mature-like state but produces neither conical cores nor infectious virions. These observations support the disassembly-reassembly pathway for core formation.


Laws can be unethical.
ICU teams have the difficult emotional burden of continuing complex life-sustaining therapy beyond the limits of what is felt to be reasonable. Among the reasons leading to a delay in the withdrawal of intensive therapy is the unwillingness or unpreparedness of the team or family members, or inadequate laws. We all have the responsibility to promote a legal framework allowing end-of-life decisions that ensure the autonomy, dignity and integrity of all citizens, in addition to the humane practice of medicine.