Behavior of eosinophil leukocytes in acute inflammation. I. Lack of dependence on adrenal function.
Acute infection is accompanied by a characteristic reduction in circulating eosinophils. This study examined the generally held assumption that the eosinopenia of infection is a manifestation of adrenal stimulation. Trichinosis, Escherichia coli pyelonephritis, and early subcutaneous pneumococcal abscess were used as experimental infections of limited severity. Trichinosis is associated with eosinophilia, but pyelonephritis and pneumococcal infection produce eosinopenia. An assay for serum corticosterone was developed that is sufficiently sensitive to be performed with the small volumes of blood obtained sequentially from individual mice. The corticosterone response to trichinosis fits the sterotyped reaction previously reported for several other bacterial, viral, and rickettsial infections. The peak concentrations of corticosterone in serum from mice with trichinosis was approximately twice normal and occurred at the onset of clinical illness. Serum corticosterone levels gradually declined to the normal range over the next several days. E. coli pyelonephritis produced a similar adrenal response, although the peak serum corticosterone caused by pyelonephritis was less than the serum corticosterone occurring during the first peak of eosinophilia during trichinosis. Infection of a subcutaneous air pouch with penumococci produced eosinopenia within 6 h after inoculation, but there was no rise in serum corticosterone during the first 12 h of the pneumococcal infection. In addition, the eosinopenic response produced by a 12-hpneumococcal abscess occurred mice adrenalectomized 1-4 days before infection with pneumococci. The eosinopenia of acute infection cannot be ascribed to adrenal stimulation.


Regulation of E2F1 activity by acetylation.
During the G(1) phase of the cell cycle, an E2F-RB complex represses transcription, via the recruitment of histone deacetylase activity. Phosphorylation of RB at the G(1)/S boundary generates a pool of 'free' E2F, which then stimulates transcription of S-phase genes. Given that E2F1 activity is stimulated by p300/CBP acetylase and repressed by an RB-associated deacetylase, we asked if E2F1 was subject to modification by acetylation. We show that the p300/CBP-associated factor P/CAF, and to a lesser extent p300/CBP itself, can acetylate E2F1 in vitro and that intracellular E2F1 is acetylated. The acetylation sites lie adjacent to the E2F1 DNA-binding domain and involve lysine residues highly conserved in E2F1, 2 and 3. Acetylation by P/CAF has three functional consequences on E2F1 activity: increased DNA-binding ability, activation potential and protein half-life. These results suggest that acetylation stimulates the functions of the non-RB bound 'free' form of E2F1. Consistent with this, we find that the RB-associated histone deacetylase can deacetylate E2F1. These results identify acetylation as a novel regulatory modification that stimulates E2F1's activation functions.


Unplanned Readmissions Following Outpatient Hand and Elbow Surgery.
Unplanned readmission following surgery is a quality metric that helps surgeons assess initiatives targeted at improving patient care. We utilized the American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP) database to determine the rates, causes, and predictors of unplanned 30-day readmissions after outpatient elective hand and elbow surgery. The ACS-NSQIP database was queried using hand-and-elbow-specific Current Procedural Terminology (CPT) codes to retrospectively identify patients who had undergone outpatient hand or elbow surgery in 2012 and 2013. Patients who required an unplanned readmission to the hospital within 30 days were compared with those who were not readmitted. Preoperative patient characteristics, intraoperative variables, complications, and mortality were compared between the cohorts. Cox proportional hazard models were used to determine independent associations with 30-day unplanned readmission. A total of 14,106 outpatient hand or elbow surgery procedures were identified between 2012 and 2013, and 169 (1.2%) of them were followed by an unplanned readmission. The leading causes of readmission included postoperative infections (19.5%), pain (4.7%), thromboembolic events (4.1%), and pulmonary complications (3.0%). The causes of approximately 1/3 of the readmissions were missing from the database, and these readmissions were likely unrelated to the principal procedure. Independent predictors of readmission included an age of 70 to 84 years (hazard ratio [HR] = 2.83, 95% confidence interval [CI] = 1.67 to 4.78), smoking (HR = 2.23, 95% CI = 1.57 to 3.18), a lower hematocrit (HR = 2.19, 95% CI = 1.38 to 3.46), renal dialysis (HR = 3.32, 95% CI = 1.60 to 6.91), and an elbow procedure (with or without a hand procedure) (HR = 2.19, 95% CI = 1.57 to 3.04). The prevalence of unplanned readmission following outpatient hand and elbow surgery is low. Several modifiable factors, including preoperative smoking and anemia, are associated with unplanned readmission. These data may be helpful in developing quality-control initiatives to target unplanned readmissions following hand and elbow procedures. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


A wavelet-based method to exploit epigenomic language in the regulatory region.
Epigenetic landscapes in the regulatory regions reflect binding condition of transcription factors and their co-factors. Identifying epigenetic condition and its variation is important in understanding condition-specific gene regulation. Computational approaches to explore complex multi-dimensional landscapes are needed. To study epigenomic condition for gene regulation, we developed a method, AWNFR, to classify epigenomic landscapes based on the detected epigenomic landscapes. Assuming mixture of Gaussians for a nucleosome, the proposed method captures the shape of histone modification and identifies potential regulatory regions in the wavelet domain. For accuracy estimation as well as enhanced computational speed, we developed a novel algorithm based on down-sampling operation and footprint in wavelet. We showed the algorithmic advantages of AWNFR using the simulated data. AWNFR identified regulatory regions more effectively and accurately than the previous approaches with the epigenome data in mouse embryonic stem cells and human lung fibroblast cells (IMR90). Based on the detected epigenomic landscapes, AWNFR classified epigenomic status and studied epigenomic codes. We studied co-occurring histone marks and showed that AWNFR captures the epigenomic variation across time. The source code and supplemental document of AWNFR are available at http://wonk.med.upenn.edu/AWNFR.


Telephone triage by nurses in primary care out-of-hours services in Norway: an evaluation study based on written case scenarios.
The use of nurses for telephone-based triage in out-of-hours services is increasing in several countries. No investigations have been carried out in Norway into the quality of decisions made by nurses regarding our priority degree system. There are three levels: acute, urgent and non-urgent. Nurses working in seven casualty clinics in out-of-hours districts in Norway (The Watchtowers) were all invited to participate in a study to assess priority grade on 20 written medical scenarios validated by an expert group. 83 nurses (response rate 76%) participated in the study. A one-out-of-five sample of the nurses assessed the same written cases after 3 months (n = 18, response rate 90%) as a test-retest assessment. Among the acute, urgent and non-urgent scenarios, 82%, 74% and 81% were correctly classified according to national guidelines. There were significant differences in the proportion of correct classifications among the casualty clinics, but neither employment percentage nor profession or work experience affected the triage decision. The mean intraobserver variability measured by the Cohen kappa was 0.61 (CI 0.52 to 0.70), and there were significant differences in kappa with employment percentage. Casualty clinics and work experience did not affect intrarater agreement. Correct classification of acute and non-urgent cases among nurses was quite high. Work experience and employment percentage did not affect triage decision. The intrarater agreement was good and about the same as in previous studies performed in other countries. Kappa increased significantly with increasing employment percentage.


Shedding Light on Keeping People in the Dark.
We want to keep hackers in the dark about our passwords and our credit card numbers. We want to keep potential eavesdroppers in the dark about our private communications with friends and business associates. This need for secrecy raises important questions in epistemology (how do we do it?) and in ethics (should we do it?). In order to answer these questions, it would be useful to have a good understanding of the concept of keeping someone in the dark. Several philosophers (e.g., Bok, 1983; Carson, 2010; Mahon, 2009; Scheppele, 1988) have analyzed this concept (or, equivalently, the concept of keeping secrets) in terms of concealing and/or withholding information. However, their analyses incorrectly exclude clear instances of keeping someone in the dark. And more important, they incorrectly focus on possible means of keeping someone in the dark rather than on what it is to keep someone in the dark. In this paper, I argue that you keep X in the dark about a proposition P if and only if you intentionally cause X not to have a true belief that P. In addition, I show how this analysis of keeping someone in the dark can be extended from a categorical belief model of epistemic states to a credence (or degree of belief) model.


Blueberries decrease cardiovascular risk factors in obese men and women with metabolic syndrome.
Among all fruits, berries have shown substantial cardio-protective benefits due to their high polyphenol content. However, investigation of their efficacy in improving features of metabolic syndrome and related cardiovascular risk factors in obesity is limited. We examined the effects of blueberry supplementation on features of metabolic syndrome, lipid peroxidation, and inflammation in obese men and women. Forty-eight participants with metabolic syndrome [4 males and 44 females; BMI: 37.8 +/- 2.3 kg/m(2); age: 50.0 +/- 3.0 y (mean +/- SE)] consumed freeze-dried blueberry beverage (50 g freeze-dried blueberries, approximately 350 g fresh blueberries) or equivalent amounts of fluids (controls, 960 mL water) daily for 8 wk in a randomized controlled trial. Anthropometric and blood pressure measurements, assessment of dietary intakes, and fasting blood draws were conducted at screening and at wk 4 and 8 of the study. The decreases in systolic and diastolic blood pressures were greater in the blueberry-supplemented group (- 6 and - 4%, respectively) than in controls (- 1.5 and - 1.2%) (P lt 0.05), whereas the serum glucose concentration and lipid profiles were not affected. The decreases in plasma oxidized LDL and serum malondialdehyde and hydroxynonenal concentrations were greater in the blueberry group (- 28 and - 17%, respectively) than in the control group (- 9 and - 9%) (P lt 0.01). Our study shows blueberries may improve selected features of metabolic syndrome and related cardiovascular risk factors at dietary achievable doses.


Time course of LPS-induced gene expression in a mouse model of genitourinary inflammation.
In this study, self-organizing map (SOM) gene cluster techniques are applied to the analysis of cDNA microarray analysis of gene expression changes occurring in the early stages of genitourinary inflammation. We determined the time course of lipopolysaccharide (LPS)-induced gene expression in experimental cystitis. Mice were euthanized 0.5, 1, 4, and 24 h after LPS instillation into the urinary bladder, and gene expression was determined using four replicate Atlas mouse cDNA expression arrays containing 588 known genes at each time point. SOM gene cluster analysis, performed without preconditions, identified functionally significant gene clusters based on the kinetics of change in gene expression. Genes were classified as follows: 1) expressed at time 0; 2) early genes (peak expression between 0.5 and 1 h); and 3) late genes (peak expression between 4 and 24 h). One gene cluster maintained a constant level of expression during the entire time period studied. In contrast, LPS treatment downregulated the expression of some genes expressed at time 0, in a cluster including transcription factors, protooncogenes, apoptosis-related proteins (cysteine protease), intracellular kinases, and growth factors. Gene upregulation in response to LPS was observed as early as 0.5 h in a cluster including the interleukin-6 (IL-6) receptor, alpha- and beta-nerve growth factor (alpha- and beta-NGF), vascular endothelial growth factor receptor-1 (VEGF R1), C-C chemokine receptor, and P-selectin. Another tight cluster of genes with marked expression at 1 h after LPS and insignificant expression at all other time points studied included the protooncogenes c-Fos, Fos-B, Fra-2, Jun-B, Jun-D, and Egr-1. Almost all interleukin genes were upregulated as early as 1 h after stimulation with LPS. Nuclear factor-kappaB (NF-kappaB) pathway genes collected in a single cluster with a peak expression 4 h after LPS stimulation. In contrast, most of the interleukin receptors and chemokine receptors presented a late peak of expression 24 h after LPS coinciding with the peak of neutrophil infiltration into the bladder wall. Selected cDNA microarray observations were confirmed by RNase protection assay. In conclusion, the cDNA array experimental approach provided a global profile of gene expression changes in bladder tissue after stimulation with LPS. SOM techniques identified functionally significant gene clusters, providing a powerful technical basis for future analysis of mechanisms of bladder inflammation.


Supracricoid laryngectomy outcomes: The Johns Hopkins experience.
To report the oncologic and functional results from our experience in performing supracricoid laryngectomy (SCL) for selected patients with laryngeal cancer. Retrospective chart review. Twenty-four consecutive patients who underwent SCL for laryngeal cancer in our institution from December 2000 to March 2006 have been reviewed. Reports of the site and extent of tumor, type of reconstruction, preoperative or postoperative radiotherapy, and the final histopathologic examination were reviewed. In addition, the reports of the preoperative examination, inpatient course, and postoperative follow-up were reviewed. A total of 24 patients were involved in the study; 19 had tumors involving the glottic region, and 5 patients had tumors involving both the glottic and supraglottic regions (transglottic). Ten patients had their SCL for postradiotherapy recurrence/persistence of disease. Eighteen patients underwent reconstruction through cricohyoidoepiglottopexy (CHEP), whereas six patients had cricohyoidopexy (CHP). Eleven patients had an arytenoid cartilage resected; 8 of 11 had CHEP, and 3 of 11 had CHP. All patients had a tracheostomy and percutaneous endoscopic gastrostomy (PEG) tube placement performed at the same time as the SCL. The median hospital stay period was 6 days. Twenty-three of 24 had successful tracheostomy tube decannulation, with a median time to decannulation of 37 days. The median time to removal of the PEG tube was 70 days. The complications with SCL were postoperative wound infection in two patients (SCL/CHP) and the need for completion total laryngectomy secondary to intractable aspiration in one patient with SCL/CHP. One patient with SCL/CHEP had a ruptured pexy and subsequently underwent a second reconstruction with successful tracheostomy and PEG tube removal. One of 24 patients is still PEG tube dependant, and he had postoperative radiotherapy. Fifteen patients underwent concurrent neck dissection. None of the patients had any local or regional recurrence, with a median follow-up period of 3 years. All final surgical margins were negative for tumor invasion. Three patients had postoperative radiotherapy, two patients because of nodal metastases in the excised lymph nodes and one because of perineural invasion on final histopathologic examination of the SCL specimen. There were no perioperative deaths. SCL with CHEP or CHP represents an effective technique that can be taught and effectively used to avoid a total laryngectomy while maintaining physiologic speech and swallowing in selected patients with advanced stage primary laryngeal cancer or recurrent/persistent laryngeal cancer after radiotherapy. There is a good functional recovery with acceptable morbidity and an excellent oncologic outcome when strict selection criteria are applied and a formal swallowing rehabilitation program is followed.


Overexpression of G6PD Represents a Potential Prognostic Factor in Clear Cell Renal Cell Carcinoma.
Glucose-6-phosphate dehydrogenase (G6PD) participates in glucose metabolism and it acts as the rate-limiting enzyme of the pentose phosphate pathway (PPP). Recently, G6PD dysregulation has been found in a variety of human cancers. Through analyzing published data in The Cancer Genome Atlas (TCGA), our pilot study indicated that G6PD mRNA expression was significantly higher in advanced Fuhrman grade in clear cell renal cell carcinoma (ccRCC). These clues promoted us to further evaluate the expression profile of G6PD and its prognostic impact in patients with ccRCC. In this study, G6PD expression levels were analyzed in 149 human ccRCC and normal tissues using immunohistochemistry. The results showed that compared with that in the normal renal samples, G6PD was found highly expressed in 51.0% of ccRCC (p<0.05). High expression of G6PD was significantly correlated to tumor extent, lymph node metastasis, Fuhrman grade, and TNM stage of ccRCC (all p<0.05). Moreover, positive G6PD expression was associated with poorer overall survival in ccRCC (p<0.001). In Cox regression analyses, high expression of G6PD also could be an independent prognostic factor for overall survival in ccRCC (p=0.007). This study suggests that overexpression of G6PD is associated with advanced disease status and therefore may become an important prognosticator for poor outcomes in ccRCC, as well as a potential therapeutic target for developing effective treatment modalities.


Multicentre study of the learning curve and surgical performance of cytoreductive surgery with intraperitoneal chemotherapy for pseudomyxoma peritonei.
The learning curves for cytoreductive surgery with intraperitoneal chemotherapy for treatment of pseudomyxoma peritonei (PMP) were explored between international centres/surgeons to identify institutional or other factors that might affect performance. Data from patients with PMP treated with the combined procedure across 33 international centres between 1993 and 2012 were analysed retrospectively. A risk-adjusted sequential probability ratio test was conducted after defining the target outcome as early oncological failure (disease progression within 2 years of treatment), an acceptable risk for the target outcome (odds ratio) of 2, and type I/II error rates of 5 per cent. The risk prediction model was elaborated and patients were evaluated sequentially for each centre/surgeon. The learning curve was considered to be overcome and proficiency achieved when the odds ratio for early oncological failure became smaller than 2. Rates of optimal cytoreduction, severe postoperative morbidity and early oncological failure were 84·4, 25·7 and 29·0 per cent respectively. The median annual centre volume was 17 (range 6-66) peritoneal malignancies. Only eight of the 33 centres and six of 47 surgeons achieved proficiency after a median of 100 (range 78-284) and 96 (86-284) procedures respectively. The most important institutional factor affecting surgical performance was centre volume. The learning curve is extremely long, so centralization and/or networking of centres is necessary to assure quality of services. One centre for every 10-15 million inhabitants would be ideal.


Nonlinear magnetotransport in interacting chiral nanotubes.
Nonlinear transport through interacting single-wall nanotubes containing a few impurities is studied theoretically. Extending the Luttinger liquid theory to incorporate trigonal warping and chirality effects, we derive the current contribution Ie even in the applied voltage V and odd in an orbital magnetic field B, which is nonzero only for chiral tubes and in the presence of interactions.


An epigenetic clock for human skeletal muscle.
Ageing is associated with DNA methylation changes in all human tissues, and epigenetic markers can estimate chronological age based on DNA methylation patterns across tissues. However, the construction of the original pan-tissue epigenetic clock did not include skeletal muscle samples and hence exhibited a strong deviation between DNA methylation and chronological age in this tissue. To address this, we developed a more accurate, muscle-specific epigenetic clock based on the genome-wide DNA methylation data of 682 skeletal muscle samples from 12 independent datasets (18-89 years old, 22% women, 99% Caucasian), all generated with Illumina HumanMethylation (HM) arrays (HM27, HM450, or HMEPIC). We also took advantage of the large number of samples to conduct an epigenome-wide association study of age-associated DNA methylation patterns in skeletal muscle. The newly developed clock uses 200 cytosine-phosphate-guanine dinucleotides to estimate chronological age in skeletal muscle, 16 of which are in common with the 353 cytosine-phosphate-guanine dinucleotides of the pan-tissue clock. The muscle clock outperformed the pan-tissue clock, with a median error of only 4.6 years across datasets (vs. 13.1 years for the pan-tissue clock, P < 0.0001) and an average correlation of ρ = 0.62 between actual and predicted age across datasets (vs. ρ = 0.51 for the pan-tissue clock). Lastly, we identified 180 differentially methylated regions with age in skeletal muscle at a false discovery rate < 0.005. However, gene set enrichment analysis did not reveal any enrichment for gene ontologies. We have developed a muscle-specific epigenetic clock that predicts age with better accuracy than the pan-tissue clock. We implemented the muscle clock in an r package called Muscle Epigenetic Age Test available on Bioconductor to estimate epigenetic age in skeletal muscle samples. This clock may prove valuable in assessing the impact of environmental factors, such as exercise and diet, on muscle-specific biological ageing processes.


The flavonoid baicalin protects against cerebrovascular dysfunction and brain inflammation in experimental heatstroke.
The present study was performed to assess the prophylactic effect of baicalin, a flavonoid compound, in an animal model of heatstroke. Anesthetized rats, immediately before the start of heat stress, were divided into two major groups and given the following: vehicle solution (1mL per kg body weight) or baicalin (10-40mg per kg body weight) intravenously. They were exposed to ambient temperature of 43 degrees C to induce heatstroke. Another group of rats was exposed to room temperature (24 degrees C) and used as normothermic controls. Their physiologic and biochemical parameters were continuously monitored. When the vehicle-pretreated rats underwent heat stress, their survival time values were found to be 20-28min. Pretreatment with intravenous doses of baicalin significantly improved survival during heatstroke (65-248min). As compared to those of normothermic controls, all vehicle-pretreated heatstroke animals displayed higher levels of core temperature, intracranial pressure, and nitric oxide metabolite (NO(2)(-)), glutamate, glycerol, lactate/pyruvate ratio, and dihydroxybenzoic acid (DHBA) in hypothalamus. In addition, both serum and hypothalamic levels of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) as well as plasma levels of creatinine, serum urea nitrogen, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and alkaline phosphatase were elevated after heatstroke onset. In contrast, all vehicle-pretreated heatstroke animals had lower levels of mean arterial pressure, cerebral perfusion pressure, cerebral blood flow, and brain PO(2). Administration of baicalin before the start of heat exposure significantly reduced the hyperthermia, intracranial hypertension, and the increased levels of NO(2)(-), glutamate, glycerol, lactate/pyruvate ratio, and DHBA in the hypothalamus that occurred during heatstroke. The heatstroke-induced increased levels of IL-1beta and TNF-alpha in both the serum and hypothalamus, and renal and hepatic dysfunction were suppressed by baicalin pretreatment. In contrast, both the serum and hypothalamic levels of IL-10 were significantly elevated by baicalin during heatstroke. We successfully demonstrated that baicalin can be used as a prophylactic agent for heatstroke. In particular, baicalin may protect against cerebrovascular dysfunction and brain inflammation in heatstroke.


Sympathetic nervous system modulates the ocular hypotensive action of MT2-melatonin receptors in normotensive rabbits.
The aim of this study was to investigate the hypotensive effect of the melatonin analogue, N-butanoyl-2-(2-methoxy-6H-isoindolo[2,1-a]indol-11-yl)ethanamine (IIK7), through MT(2)-melatonin receptors and the involvement of the sympathetic nervous system in this action in New Zealand rabbit eyes. The topical application of melatonin or IIK7 produced a reduction in intraocular pressure of 20.2 +/- 5.3% and 38.5 +/- 3.2% respectively. This effect was concentration-dependent; it was blocked by selective MT(2) receptor antagonists and was severely diminished after chemical sympathectomy. Immunohistochemistry and western blot analysis showed the ciliary processes as the site of this action and no co-localization of MT(2)-melatonin receptor with the sympathetic nervous system was observed. The beta-adrenergic agonists, terbutaline and salbutamol, potentiated the hypotensive effect of IIK7 reducing intraocular pressure (IOP) 41.75 +/- 4.26% and 44.7 +/- 5.6% respectively. Also, IIK7 in presence of the nonspecific protein phosphatase inhibitor okadaic acid, lowered IOP 32.2 +/- 4.5% and in presence of forskolin plus 3-isobutyl-1-methylxanthine decreased IOP in 32.2 +/- 5.47%. These data suggest that the melatonin agonist IIK7 reduces intraocular pressure by acting through MT(2)-melatonin receptors presumably decreasing aqueous humour formation. Also, in the presence of beta-adrenoceptor agonists MT(2)-melatonin receptors activity increase their ability to reduce IOP.


A review of bovine fasciolosis and other trematode infections in Nigeria.
Trematode infections cause serious economic losses to livestock worldwide. Global production losses due to fasciolosis alone exceed US$3 billion annually. Many trematode infections are also zoonotic and thus a public health concern. The World Health Organization has estimated that about 56 million people worldwide are infected by at least one zoonotic trematode species, and up to 750 million people are at risk of infection. Fasciolosis caused by the fluke Fasciola gigantica is endemic in Nigeria and is one of the most common causes of liver condemnation in abattoirs. Total cattle losses from Fasciola infection in Nigeria have been estimated to cost £32.5 million. Other trematode infections of cattle, including paramphistomosis, dicrocoeliasis and schistosomiasis, have all been reported in various parts of Nigeria, with varying prevalence. Most publications on trematode infections are limited to Nigerian local and national journals, with very few international reports. This paper therefore summarized the current data on distribution, control and zoonotic trematode infections in Nigeria and other African countries. We also identified research gaps and made recommendations for future research and areas for funding for policy/planning.


Preclinical Assessment of a Gene-Editing Approach in a Mouse Model of Mitochondrial Neurogastrointestinal Encephalomyopathy.
Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is a rare disease caused by recessive mutations in the TYMP gene, which encodes the enzyme thymidine phosphorylase (TP). In this study, the efficient integration of a TYMP transgene into introns of the Tymp and Alb loci of hepatocytes in a murine model of MNGIE was achieved by the coordinated delivery and activity of CRISPR/Cas9 and a TYMP cDNA. CRISPR/Cas9 was delivered either as mRNA using lipid nanoparticle (LNP) or polymeric nanoparticle, respectively, or in an AAV2/8 viral vector; the latter was also used to package the TYMP cDNA. Insertion of the cDNA template downstream of the Tymp and Alb promoters ensured transgene expression. The best in vivo results were obtained using LNP carrying the CRISPR/Cas9 mRNAs. Treated mice showed a consistent long-term (1 year) reduction in plasma nucleoside (thymidine and deoxyuridine) levels that correlated with the presence of TYMP mRNA and functional enzyme in liver cells. In mice with an edited Alb locus, the transgene produced a hybrid Alb-hTP protein that was secreted, with supraphysiological levels of TP activity detected in the plasma. Equivalent results were obtained in mice edited at the Tymp locus. Finally, some degree of gene editing was found in animals treated only with AAV vectors containing the DNA templates, in the absence of nucleases, although there was no impact on plasma nucleoside levels. Overall, these results demonstrate the feasibility of liver-directed genome editing in the long-term correction of MNGIE, with several advantages over other methods.


Digit Force Controls and Corresponding Brain Activities in Finger Pressing Performance: A Comparison Between Older Adults and Young Individuals.
This study aims toward an investigation and comparison of the digital force control and the brain activities of older adults and young groups during digital pressing tasks. A total of 15 young and 15 older adults were asked to perform force ramp tasks at different force levels with a custom pressing system. Near-infrared spectroscopy was used to collect the brain activities in the prefrontal cortex and primary motor area. The results showed that the force independence and hand function of the older adults were worse than that of the young adults. The cortical activations in the older adults were higher than those in the young group during the tasks. A significant hemodynamic between-group response and mild negative correlations between brain activation and force independence ability were found. Older adults showed poor force independence ability and manual dexterity and required additional brain activity to compensate for the degeneration.


Steroid hormones: effects on adenyl cyclase activity and adenosine 3',5'-momophosphate in target tissues.
The adenyl cyclases of chick oviduct and rat prostate were not stimulated by estrogen and testosterone, respectively, suggesting that growth and differentiation of these target tissues are not mediated by adenosine 3',5'-monophosphate. Estrogen acutely activated adenyl cyclase in the castrate rat uterus, but this was prevented by administration of DL-propranolol, suggesting that the effect was mediated by catecholamines. Progesterone produced a delayed stimulation of oviduct adenyl cyclase preceding and concomitant with the induction of synthesis of avidin.


Extracellular Alkaline Lipase from a Novel Fungus Curvularia sp. DHE 5: Optimisation of Physicochemical Parameters, Partial Purification and Characterisation.
Thirty isolated fungal strains were screened for lipase production using Phenol Red plates, containing tributyrin as lipidic substrate, and a novel fungus identified genetically as Curvularia sp. DHE 5 was found as the most prominent strain. Various agro-industrial substrates were evaluated as inert supports for lipase production in solid-state fermentation. The highest yield of lipase ((83.4±2.2) U/g on dry mass basis) was reported with wheat bran medium after seven days of fermentation at pH=7.0, temperature of 30 °C, 70% moisture content, inoculum size of 1.27·107 spore/mL and 2% olive oil as an inducer. Supplementation of the medium with 0.05% KCl as an ion source further increased lipase production to (88.9±1.2) U/g on dry mass basis. The enzyme was partially purified through ammonium sulphate fractionation (40%) followed by dialysis, and its optimum pH and temperature were reported at 8.0 and 50 °C, respectively, with remarkable pH and thermal stability.


Long-term effects of marijuana use on the brain.
Questions surrounding the effects of chronic marijuana use on brain structure continue to increase. To date, however, findings remain inconclusive. In this comprehensive study that aimed to characterize brain alterations associated with chronic marijuana use, we measured gray matter (GM) volume via structural MRI across the whole brain by using voxel-based morphology, synchrony among abnormal GM regions during resting state via functional connectivity MRI, and white matter integrity (i.e., structural connectivity) between the abnormal GM regions via diffusion tensor imaging in 48 marijuana users and 62 age- and sex-matched nonusing controls. The results showed that compared with controls, marijuana users had significantly less bilateral orbitofrontal gyri volume, higher functional connectivity in the orbitofrontal cortex (OFC) network, and higher structural connectivity in tracts that innervate the OFC (forceps minor) as measured by fractional anisotropy (FA). Increased OFC functional connectivity in marijuana users was associated with earlier age of onset. Lastly, a quadratic trend was observed suggesting that the FA of the forceps minor tract initially increased following regular marijuana use but decreased with protracted regular use. This pattern may indicate differential effects of initial and chronic marijuana use that may reflect complex neuroadaptive processes in response to marijuana use. Despite the observed age of onset effects, longitudinal studies are needed to determine causality of these effects.


Decamethyltitanocene hydride intermediates in the hydrogenation of the corresponding titanocene-(η2-ethene) or (η2-alkyne) complexes and the effects of bulkier auxiliary ligands.
1H NMR studies of reactions of titanocene [Cp*2Ti] (Cp* = η5-C5Me5) and its derivatives [Cp*(η5:η1-C5Me4CH2)TiMe] and [Cp*2Ti(η2-CH2[double bond, length as m-dash]CH2)] with excess dihydrogen at room temperature and pressures lower than 1 bar revealed the formation of dihydride [Cp*2TiH2] (1) and the concurrent liberation of either methane or ethane, depending on the organometallic reactant. The subsequent slow decay of 1 yielding [Cp*2TiH] (2) was mediated by titanocene formed in situ and controlled by hydrogen pressure. The crystalline products obtained by evaporating a hexane solution of fresh [Cp*2Ti] in the presence of hydrogen contained crystals having either two independent molecules of 1 in the asymmetric part of the unit cell or cocrystals consisting of 1 and [Cp*2Ti] in a 2 : 1 ratio. Hydrogenation of alkyne complexes [Cp*2Ti(η2-R1C[triple bond, length as m-dash]CR2)] (R1 = R2 = Me or Et) performed at room temperature afforded alkanes R1CH2CH2R2, and after removing hydrogen, 2 was formed in quantitative yields. For alkyne complexes containing bulkier substituent(s) R1 = Me or Ph, R2 = SiMe3, and R1 = R2 = Ph or SiMe3, successful hydrogenation required the application of increased temperatures (70-80 °C) and prolonged reaction times, in particular for bis(trimethylsilyl)acetylene. Under these conditions, no transient 1 was detected during the formation of 2. The bulkier auxiliary ligands η5-C5Me4tBu and η5-C5Me4SiMe3 did not hinder the addition of dihydrogen to the corresponding titanocenes [(η5-C5Me4tBu)2Ti] and [(η5-C5Me4SiMe3)2Ti] yielding [(η5-C5Me4tBu)2TiH2] (3) and [(η5-C5Me4SiMe3)2TiH2] (4), respectively. In contrast to 1, the dihydride 4 did not decay with the formation of titanocene monohydride, but dissociated to titanocene upon dihydrogen removal. The monohydrides [(η5-C5Me4tBu)2TiH] (5) and [(η5-C5Me4SiMe3)2TiH] (6) were obtained by insertion of dihydrogen into the intramolecular titanium-methylene σ-bond in compounds [(η5-C5Me4tBu)(η5:η1-C5Me4CMe2CH2)Ti] and [(η5-C5Me4SiMe3)(η5:η1-C5Me4SiMe2CH2)Ti], respectively. The steric influence of the auxiliary ligands became clear from the nature of the products obtained by reacting 5 and 6 with butadiene. They appeared to be the exclusively σ-bonded η1-but-2-enyl titanocenes (7) and (8), instead of the common π-bonded derivatives formed for the sterically less congested titanocenes, including [Cp*2Ti(η3-(1-methylallyl))] (9). The molecular structure optimized by DFT for compound 1 acquired a distinctly lower total energy than the analogously optimized complex with a coordinated dihydrogen [Cp*2Ti(η2-H2)]. The stabilization energies of binding the hydride ligands to the bent titanocenes were estimated from counterpoise computations; they showed a decrease in the order 1 (-132.70 kJ mol-1), 3 (-121.11 kJ mol-1), and 4 (-112.35 kJ mol-1), in accordance with the more facile dihydrogen dissociation.


The brown midrib3 (bm3) mutation in maize occurs in the gene encoding caffeic acid O-methyltransferase.
The brown midrib mutations are among the earliest described in maize. Plants containing a brown midrib mutation exhibit a reddish brown pigmentation of the leaf midrib starting when there are four to six leaves. These mutations are known to alter lignin composition and digestibility of plants and therefore constitute prime candidates in the breeding of silage maize. Here, we show that two independent brown midrib3 (bm3) mutations have resulted from structural changes in the COMT gene, which encodes the enzyme O-methyltransferase (COMT; EC 22.214.171.124), involved in lignin biosynthesis. Our results indicate that the bm3-1 allele (the reference mutant allele) has arisen from an insertional event producing a COMT mRNA altered in both size and amount. By sequencing a COMT cDNA clone obtained from bm3-1 maize, a retrotransposon with homology to the B5 element has been found to be inserted near the junction of the 3' coding region of the COMT gene intron. The second bm3 allele, bm3-2, has resulted from a deletion of part of the COMT gene. These alterations of the COMT gene were confirmed by DNA gel blot and polymerase chain reaction amplification analyses. These results clearly demonstrate that mutations at the COMT gene give a brown midrib3 phenotype. Thus, the gene genetically recognized as bm3 is the same as the one coding for COMT.


Age-specific interval breast cancers in Japan: estimation of the proper sensitivity of screening using a population-based cancer registry.
The age-specific sensitivity of a screening program was investigated using a population-based cancer registry as a source of false-negative cancer cases. A population-based screening program for breast cancer was run using either clinical breast examinations (CBE) alone or mammography combined with CBE in the Miyagi Prefecture from 1997 to 2002. Interval cancers were newly identified by linking the screening records to the population-based cancer registry to estimate the number of false-negative cases of screening program. Among 112 071 women screened by mammography combined with CBE, the number of detected cancers, false-negative cases and the sensitivity were 289, 22 and 92.9%, respectively, based on the reports from participating municipalities. The number of newly found false-negative cases and corrected sensitivity when using the registry were 34 and 83.8%, respectively. In detected cancers, the sensitivity of screening by mammography combined with CBE in women ranging from 40 to 49 years of age based on a population-based cancer registry was much lower than that in women 50-59 and 60-69 years of age (40-49: 18, 71.4%, 50-59: 19, 85.8%, 60-69: 19, 87.2%). These data suggest that the accurate outcome of an evaluation of breast cancer screening must include the use of a population-based cancer registry for detecting false-negative cases. Screening by mammography combined with CBE may therefore not be sufficiently sensitive for women ranging from 40 to 49 years of age.


Evolution and Phylogeny of Large DNA Viruses, Mimiviridae and Phycodnaviridae Including Newly Characterized Heterosigma akashiwo Virus.
Nucleocytoplasmic DNA viruses are a large group of viruses that harbor double-stranded DNA genomes with sizes of several 100 kbp, challenging the traditional concept of viruses as small, simple 'organisms at the edge of life.' The most intriguing questions about them may be their origin and evolution, which have yielded the variety we see today. Specifically, the phyletic relationship between two giant dsDNA virus families that are presumed to be close, Mimiviridae, which infect Acanthamoeba, and Phycodnaviridae, which infect algae, is still obscure and needs to be clarified by in-depth analysis. Here, we studied Mimiviridae-Phycodnaviridae phylogeny including the newly identified Heterosigma akashiwo virus strain HaV53. Gene-to-gene comparison of HaV53 with other giant dsDNA viruses showed that only a small proportion of HaV53 genes show similarities with the others, revealing its uniqueness among Phycodnaviridae. Phylogenetic/genomic analysis of Phycodnaviridae including HaV53 revealed that the family can be classified into four distinctive subfamilies, namely, Megaviridae (Mimivirus-like), Chlorovirus-type, and Coccolitho/Phaeovirus-type groups, and HaV53 independent of the other three groups. Several orthologs found in specific subfamilies while absent from the others were identified, providing potential family marker genes. Finally, reconstruction of the evolutionary history of Phycodnaviridae and Mimiviridae revealed that these viruses are descended from a common ancestor with a small set of genes and reached their current diversity by differentially acquiring gene sets during the course of evolution. Our study illustrates the phylogeny and evolution of Mimiviridae-Phycodnaviridae and proposes classifications that better represent phyletic relationships among the family members.


Diabetic persons with foot ulcers and their perceptions of hyperbaric oxygen chamber therapy.
To elucidate how diabetic patients with limb-threatening foot lesions perceive and evaluate content and organisation of treatment in a multi-place hyperbaric oxygen chamber. To our knowledge there are no patients' evaluations of diabetes care in a high-technology area like the hyperbaric oxygen chamber. The burden on persons with diabetic foot complications might be increased if adjuvant therapy with hyperbaric oxygen therapy (HBOT) within a locked airtight vessel is given. Explorative study. Participants were included in the HODFU study, a prospective randomised double-blind study, designed to evaluate whether HBOT heals more chronic foot ulcers than placebo treatment with hyperbaric air. Six females and 13 males, aged 44-83 years (median 70), with diabetic foot ulcers, participated. Focus-group interviews by an external evaluator. Management was perceived as well-functioning with competent staff delivering quick treatment in a positive manner and in good co-operation. HBOT sessions, in groups, were described as unproblematic and pleasant, through sharing experiences with others, although time-consuming and tiring. Recognising the responsible physician and communication with other physicians in the health-care chain was perceived as problematic. Placebo treatment, when given, did not reveal any problems; many perceived HBOT as the last resort and respondents had a negative view of future health and expressed fears of new wounds and amputation. From patients' perspective HBOT in the delivered health-care model was perceived as well-functioning, taking into consideration both technical and relational aspects of care in this high-technology environment. Communication with the patient and between different care givers, with a consistent message given and information about who is responsible and to whom one should turn, wherever treated, is the most crucial aspect of the model. Future fears need to be recognised and group interaction can be encouraged to share the burden of disease.


Antireflection surfaces in silicon using binary optics technology.
Binary optics processing methods were applied to a silicon substrate to generate an array of small pillars in order to enhance transmission. The volume fraction of the silicon in the pillars was chosen to simulate a single homogeneous antireflection layer, and the pillar height was targeted to be a quarter-wave thickness. A mask was generated, using a graphics computer-aided design system; reactive-ion etching was used to generate the pillars. An improvement in long-wavelength infrared transmission is observed, with diffraction and scattering dominating at shorter wavelengths.


[Subcutaneous heparin during the first trimesters of pregnancy in women with prosthetic heart valves].
Oral anticoagulation therapy in pregnant women with prosthetic heart valves is associated with a greater risk of abortion, fetal malformations and thromboembolic complications. To assess the use of subcutaneous heparin in women with pregnancies of less than 12 weeks as an alternative to oral anticoagulation therapy. Pregnant women were admitted to the hospital and coumarinics were replaced with not fractionated subcutaneous heparin in a dose of 10,000 IU every 12 hours, aiming at prolonging partial thromboplastin time, 2 or 2 1/2 times. During the hospitalization period, women were taught about the heparin injection technique. Coumarinics were restarted after week 13 of pregnancy until 10 days prior to the delivery date in which women were again admitted to the hospital and intravenous heparin was used until the delivery. Between 1991 and 1997, this protocol was used during ten pregnancies in seven women aged 19 to 36 years old. Five had a Starr-Edwards prosthesis, one had a mitral Björk-Shilley prosthesis and one, a double prosthesis (mitral Starr-Machi and aortic St Jude). Subcutaneous heparin was started in the fifth week of pregnancy in 1 case, in the sixth week in seven and in the seventh week in two. There was no maternal mortality and one transient ischemic attack without sequelae. One non compliant patient had a Björk Shilley prosthetic valve dysfunction that required a valve replacement at the twelfth week of pregnancy and she had a spontaneous abortion at week 15. Other patient had a pneumonia at week 37 and gave birth to a stillbirth. There were no fetal malformations. This therapeutic protocol can be used among Chilean patients, but must be restricted to compliant women and a strict surveillance must be maintained.


Effect of selective noradrenergic denervation on noradrenaline content and [3H]dopamine release in rat nucleus accumbens slices.
DSP4 (N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine) treatment (50 mg/kg i.p., 10 days previously) significantly decreased the noradrenaline (NA) content of the rostral part of the nucleus accumbens. The medial and caudal areas were not affected. The nucleus accumbens appears to receive noradrenergic innervation predominantly from subcoeruleus nuclei of the pons-medulla while the locus coeruleus neurons project to the rostral area. The isoproterenol-induced enhancement of the K+-evoked release of [3H]dopamine (DA) was not affected by DSP4 treatment. Noradrenergic denervation does not appear to have been sufficient to cause up-regulation of postsynaptic beta-adrenoceptors.


Non-small cell lung cancer: current treatment and future advances.
Lung cancer has a poor prognosis; over half of people diagnosed with lung cancer die within one year of diagnosis and the 5-year survival is less than 18%. Non-small cell lung cancer (NSCLC) accounts for the majority of all lung cancer cases. Risk factors for developing NSCLC have been identified, with cigarette smoking being a major factor along with other environmental and genetic risk factors. Depending on the staging of lung cancer, patients are eligible for certain treatments ranging from surgery to radiation to chemotherapy as well as targeted therapy. With the advancement of genetics and biomarkers testing, specific mutations have been identified to better target treatment for individual patients. This review discusses current treatments including surgery, chemotherapy, radiotherapy, and immunotherapy as well as how biomarker testing has helped improve survival in patients with NSCLC.


Functionalization of any substrate using covalently modified large area CVD graphene.
We report a novel route for the functionalization of any substrates, including chemically inert substrates. CVD grown graphene is electrochemically functionalized with p-(N-maleimido)phenyl residues and consecutively transferred to various substrates. The transfer process is shown to be without noticeable loss. The functional layer exhibits a thickness of appx. 4.5 nm.


[Safety performance of rivaroxaban versus warfarin in patients with atrial fibrillation and advanced chro-nic kidney disease].
Aim To evaluate safety of using rivaroxaban in patients with stage 4 chronic kidney disease (CKD) or transient, stable decline of glomerular filtration rate (GFR) to 15-29 ml /min / 1.73 m2 in the presence of atrial fibrillation (AF).Material and methods This multicenter prospective, randomized study included patients admitted to cardiology departments from 2017 through 2019. Of 10 224 admitted patients 109 (3 %) patients with AF and stage 4 CKD or a stable decline of GFR to 15-29 ml /min / 1.73 m2 were randomized at 2:1 ratio to the rivaroxaban 15 mg /day (n=73) treatment group or to the warfarin treatment group (n=36). The primary endpoint was development of BARC and ISTH major, minor, and clinically relevant minor bleeding. Mean follow-up duration was 18 months.Results Patients receiving warfarin had a significantly higher incidence of BARC (n=26 (72.2 %) vs. n=31 (42.4 %), р<0.01) and ISTH (n=22 (61.1 %) vs. n=27 (36.9 %), p<0.01) minor bleeding and all ISTH clinically relevant (minor clinically relevant and major bleedings) n=10 (27.7 %) vs. n=8 (10.9 %), р=0.03]. The number of repeated hospitalizations was 65 (43% of patients) in the rivaroxaban treatment group and 27 (48% of patients) in the warfarin treatment group (р=0.57), including 24 (36.9 %) and 11 (40.7 %) emergency admissions in the rivaroxaban and warfarin treatment groups, respectively (р=0.96). Significant improvement of changes in creatinine clearance and GFR (by CKD-EPI and Cockroft-Gault) was observed in the rivaroxaban treatment group.Conclusion The study provided evidence for a more beneficial safety profile of rivaroxaban compared to warfarin in patients with AF and advanced CKD.


Acute response of urine pH following ammonium chloride administration to dogs.
To test the acidifying ability of the distal portion of the nephrons in healthy dogs, 0.2 g of NH4Cl/kg of body weight was given PO. Samples for venous blood gas analysis and urine pH were taken hourly for 6 hours. Systemic acidemia developed, as evidenced by a statistically significant (P less than 0.05) decrease in blood pH 1 hour after NH4Cl administration. Four hours after administration, mean urine pH decreased to a low of 5.16 +/- 0.1 and was less than 5.5 3 hours after administration. Changes in urine pH 2 hours after administration were statistically significant (P less than 0.05). In human beings, NH4Cl loading is used to detect patients with distal renal tubular acidosis (defective hydrogen ion secretion by the distal nephrons) and normal acid/base values. Distal renal tubular acidosis is diagnosed if urine pH fails to decrease to less than 5.5 after NH4Cl administration. On the basis of the findings of this study, a similar value would be valid for dogs.


Arabidopsis late blight: infection of a nonhost plant by Albugo laibachii enables full colonization by Phytophthora infestans.
The oomycete pathogen Phytophthora infestans causes potato late blight, and as a potato and tomato specialist pathogen, is seemingly poorly adapted to infect plants outside the Solanaceae. Here, we report the unexpected finding that P. infestans can infect Arabidopsis thaliana when another oomycete pathogen, Albugo laibachii, has colonized the host plant. The behaviour and speed of P. infestans infection in Arabidopsis pre-infected with A. laibachii resemble P. infestans infection of susceptible potato plants. Transcriptional profiling of P. infestans genes during infection revealed a significant overlap in the sets of secreted-protein genes that are induced in P. infestans upon colonization of potato and susceptible Arabidopsis, suggesting major similarities in P. infestans gene expression dynamics on the two plant species. Furthermore, we found haustoria of A. laibachii and P. infestans within the same Arabidopsis cells. This Arabidopsis-A. laibachii-P. infestans tripartite interaction opens up various possibilities to dissect the molecular mechanisms of P. infestans infection and the processes occurring in co-infected Arabidopsis cells.


Spatial access inequities and childhood immunisation uptake in Kenya.
Poor access to immunisation services remains a major barrier to achieving equity and expanding vaccination coverage in many sub-Saharan African countries. In Kenya, the extent to which spatial access affects immunisation coverage is not well understood. The aim of this study was to quantify spatial accessibility to immunising health facilities and determine its influence on immunisation uptake in Kenya while controlling for potential confounders. Spatial databases of immunising facilities, road network, land use and elevation were used within a cost friction algorithim to estimate the travel time to immunising health facilities. Two travel scenarios were evaluated; (1) Walking only and (2) Optimistic scenario combining walking and motorized transport. Mean travel time to health facilities and proportions of the total population living within 1-h to the nearest immunising health facility were computed. Data from a nationally representative cross-sectional survey (KDHS 2014), was used to estimate the effect of mean travel time at survey cluster units for both fully immunised status and third dose of diphtheria-tetanus-pertussis (DPT3) vaccine using multi-level logistic regression models. Nationally, the mean travel time to immunising health facilities was 63 and 40 min using the walking and the optimistic travel scenarios respectively. Seventy five percent of the total population were within one-hour of walking to an immunising health facility while 93% were within one-hour considering the optimistic scenario. There were substantial variations across the country with 62%(29/47) and 34%(16/47) of the counties with < 90% of the population within one-hour from an immunising health facility using scenarios 1 and 2 respectively. Travel times > 1-h were significantly associated with low immunisation coverage in the univariate analysis for both fully immunised status and DPT3 vaccine. Children living more than 2-h were significantly less likely to be fully immunised [AOR:0.56(0.33-0.94) and receive DPT3 [AOR:0.51(0.21-0.92) after controlling for household wealth, mother's highest education level, parity and urban/rural residence. Travel time to immunising health facilities is a barrier to uptake of childhood vaccines in regions with suboptimal accessibility (> 2-h). Strategies that address access barriers in the hardest to reach communities are needed to enhance equitable access to immunisation services in Kenya.


Genotyping and Whole-Genome Resequencing of Welsh Sheep Breeds Reveal Candidate Genes and Variants for Adaptation to Local Environment and Socioeconomic Traits.
Advances in genetic tools applied to livestock breeding has prompted research into the previously neglected breeds adapted to harsh local environments. One such group is the Welsh mountain sheep breeds, which can be farmed at altitudes of 300 m above sea level but are considered to have a low productive value because of their poor wool quality and small carcass size. This is contrary to the lowland breeds which are more suited to wool and meat production qualities, but do not fare well on upland pasture. Herein, medium-density genotyping data from 317 individuals representing 15 Welsh sheep breeds were used alongside the whole-genome resequencing data of 14 breeds from the same set to scan for the signatures of selection and candidate genetic variants using haplotype- and SNP-based approaches. Haplotype-based selection scan performed on the genotyping data pointed to a strong selection in the regions of GBA3, PPARGC1A, APOB, and PPP1R16B genes in the upland breeds, and RNF24, PANK2, and MUC15 in the lowland breeds. SNP-based selection scan performed on the resequencing data pointed to the missense mutations under putative selection relating to a local adaptation in the upland breeds with functions such as angiogenesis (VASH1), anti-oxidation (RWDD1), cell stress (HSPA5), membrane transport (ABCA13 and SLC22A7), and insulin signaling (PTPN1 and GIGFY1). By contrast, genes containing candidate missense mutations in the lowland breeds are related to cell cycle (CDK5RAP2), cell adhesion (CDHR3), and coat color (MC1R). We found new variants in genes with potentially functional consequences to the adaptation of local sheep to their environments in Wales. Knowledge of these variations is important for improving the adaptative qualities of UK and world sheep breeds through a marker-assisted selection.


Optical heterodyne sensor using the Goos-Hänchen shift.
A new sensing scheme, capable of measuring various quantities using the Goos-Häanchen shift in the polarization phase domain, is proposed. The phase retardation between s and p polarizations on total reflection is enhanced by using multiple reflections in a plane-parallel transparent plate. This phase shift can be sensitively and stably detected using an in-line heterodyne method. As an application of this method, a displacement sensor utilizing the deflection of a light beam by a lens is developed. This displacement sensor has a resolution of 60 nm over a dynamic range of 120 microm. Good agreement between theoretical and experimental results is shown.


Wnt-dependent regulation of the E-cadherin repressor snail.
Down-regulation of E-cadherin marks the initiation of the epithelial-mesenchymal transition, a process exploited by invasive cancer cells. The zinc finger transcription factor, Snail, functions as a potent repressor of E-cadherin expression that can, acting alone or in concert with the Wnt/beta-catenin/T cell factor axis, induce an epithelial-mesenchymal transition. Although mechanisms that coordinate signaling events initiated by Snail and Wnt remain undefined, we demonstrate that Snail displays beta-catenin-like canonical motifs that support its GSK3beta-dependent phosphorylation, beta-TrCP-directed ubiquitination, and proteasomal degradation. Accordingly, Wnt signaling inhibits Snail phosphorylation and consequently increases Snail protein levels and activity while driving an in vivo epithelial-mesenchymal transition that is suppressed following Snail knockdown. These findings define a potential mechanism whereby Wnt signaling stabilizes Snail and beta-catenin proteins in tandem fashion so as to cooperatively engage transcriptional programs that control an epithelial-mesenchymal transition.


Chrono-proteomics of human saliva: variations of the salivary proteome during human development.
An important contribution to the variability of any proteome is given by the time dimension that should be carefully considered to define physiological modifications. To this purpose, whole saliva proteome was investigated in a wide age range. Whole saliva was collected from 17 preterm newborns with a postconceptional age at birth of 178-217 days. In these subjects sample collection was performed serially starting immediately after birth and within about 1 year follow-up, gathering a total of 111 specimens. Furthermore, whole saliva was collected from 182 subjects aged between 0 and 17 years and from 23 adults aged between 27 and 57 years. The naturally occurring intact salivary proteome of the 316 samples was analyzed by low- and high-resolution HPLC-ESI-MS platforms. Proteins peculiar of the adults appeared in saliva with different time courses during human development. Acidic proline-rich proteins encoded by PRH2 locus and glycosylated basic proline-rich proteins encoded by PRB3 locus appeared following 180 days of postconceptional age, followed at 7 months (±2 weeks) by histatin 1, statherin, and P-B peptide. The other histatins and acidic proline-rich proteins encoded by PRH1 locus appeared in whole saliva of babies from 1 to 3 weeks after the normal term of delivery, S-type cystatins appeared at 1 year (±3 months), and basic proline-rich proteins appeared at 4 years (±1 year) of age. All of the proteinases involved in the maturation of salivary proteins were more active in preterm than in at-term newborns, on the basis of the truncated forms detected. The activity of the Fam20C kinase, involved in the phosphorylation of various proteins, started around 180 days of postconceptional age, slowly increased reaching values comparable to adults at about 2 years (±6 months) of age. Instead, MAPK14 involved in the phosphorylation of S100A9 was fully active since birth also in preterm newborns.


Biomarkers and Noncalcified Coronary Artery Plaque Progression in Older Men Treated With Testosterone.
Recent results from the Cardiovascular Trial of the Testosterone Trials showed that testosterone treatment of older men with low testosterone was associated with greater progression of noncalcified plaque (NCP). We evaluated the effect of anthropometric measures and cardiovascular biomarkers on plaque progression in individuals in the Testosterone Trial. The Cardiovascular part of the trial included 170 men aged 65 years or older with low testosterone. Participants received testosterone gel or placebo gel for 12 months. The primary outcome was change in NCP volume from baseline to 12 months, as determined by coronary computed tomography angiography (CCTA). We assayed several markers of cardiovascular risk and analyzed each marker individually in a model as predictive variables and change in NCP as the dependent variable. Of 170 enrollees, 138 (73 testosterone, 65 placebo) completed the study and were available for the primary analysis. Of 10 markers evaluated, none showed a significant association with the change in NCP volume, but a significant interaction between treatment assignment and waist-hip ratio (WHR) (P = 0.0014) indicated that this variable impacted the testosterone effect on NCP volume. The statistical model indicated that for every 0.1 change in the WHR, the testosterone-induced 12-month change in NCP volume increased by 26.96 mm3 (95% confidence interval, 7.72-46.20). Among older men with low testosterone treated for 1 year, greater WHR was associated with greater NCP progression, as measured by CCTA. Other biomarkers and anthropometric measures did not show statistically significant association with plaque progression.


Photoselective Vaporization of the Prostate Using 120 W High Performance System: A Prospective Evaluation of Results Over 2 Years.
This study evaluated the safety and efficacy of photoselective vaporization of prostate using the 120 W High Performance System for management of symptomatic benign prostatic hyperplasia (BPH). We prospectively obtained data from 229 patients between January 2009 and December 2012. We evaluated the serum prostate specific antigen level, International Prostate Symptom Score (IPSS), prostate volume; maximum urine flow rate (Qmax); and postvoid residual urine volume (PVR) in the patients at presentation and on follow-up at 1, 6, 12, and 24 months. The mean duration of the surgery, energy used, hospital stay, and intra- and postoperative complications were assessed. The mean age of the patients was 71.2 ± 9.6 years, and the mean preoperative size of the prostate was 59.41 ± 28.1 mL. The mean duration of the surgery was 47.35 ± 16.14 min, and the mean energy use was 184.39 ± 101.3 kJ. The mean time to removal of the urinary catheter was 21.45 ± 11.06 h, while the mean duration of hospital stay was 24.82 ± 11.5 h. The IPSS declined and mean Qmax increased by over twofold within the first month. The PVR also declined significantly in all groups up to 6 months after the surgery and increased slightly thereafter. Urinary urgency and incontinence occurred in two patients, while four patients developed urinary strictures. Between 12 and 24 months after the surgery, four patients underwent repeat surgery. Our findings show that photoselective vaporization is safe and effective for the management of BPH and resulted in few complications. It yielded improvements in all parameters that were sustained even up to 2 years after the surgery.


Clinical trials treating focal segmental glomerulosclerosis should measure patient quality of life.
Optimal therapy of patients with steroid-resistant primary focal segmental glomerulosclerosis (FSGS) remains controversial. This report describes the initial study design, baseline characteristics, and quality of life of patients enrolled in the FSGS Clinical Trial, a large multicenter randomized study of this glomerulopathy comparing a 12-month regimen of cyclosporine to the combination of mycophenolate mofetil and oral dexamethasone. Patients with age ranging 2-40 years, with an estimated glomerular filtration rate > 40 ml/min per 1.73 m², a first morning urine protein-to-creatinine ratio over one, and resistant to corticosteroids were eligible. The primary outcome was complete or partial remission of proteinuria over 52 weeks after randomization. In all, 192 patients were screened, of whom 138 were randomized for treatment. Ethnic distributions were 53 black, 78 white, and 7 other. By self- or parent-proxy reporting, 26 of the 138 patients were identified as Hispanic. The baseline glomerular filtration rate was 112.4 (76.5, 180.0) ml/min per 1.73 m², and urine protein was 4.0 (2.1, 5.3) g/g. Overall, the quality of life of the patients with FSGS was lower than healthy controls and similar to that of patients with end-stage renal disease. Thus, the impact of FSGS on quality of life is significant and this measurement should be included in all trials.


Shape changes of the basal ganglia and thalamus in Alzheimer's disease: a three-year longitudinal study.
A large number of Alzheimer's disease (AD) studies have focused on medial temporal and cortical atrophy, while changes in the basal ganglia or thalamus have received less attention. The aim of this study was to investigate the existence of progressive topographical shape changes in the basal ganglia (caudate nucleus, putamen, and globus pallidus) and thalamus concurrent with AD disease progression over three years. This study also examined whether declines in volumes of the basal ganglia or thalamus might be responsible for cognitive decline in patients with AD. Thirty-six patients with early stage AD and 14 normal control subjects were prospectively recruited for this study. All subjects were assessed with neuropsychological tests and MRI at baseline and Years 1 and 3. A longitudinal shape analysis of the basal ganglia and thalamus was performed by employing a boundary surface-based shape analysis method. AD patients exhibited specific regional atrophy in the right caudate nucleus and the bilateral putamen at baseline, and as the disease progressed, regional atrophic changes in the left caudate nucleus were found to conform to a distinct topography after controlling the total brain volume. Volumetric decline of the caudate nucleus and putamen correlated with cognitive decline in frontal function after controlling for age, gender, education, follow-up years, and total brain volume changes. Our findings suggest that shape changes of the basal ganglia occurred regardless of whole brain atrophy as AD progressed and were also responsible for cognitive decline that was observed from the frontal function tests.


Genetic analysis of chromosome pairing, recombination, and cell cycle control during first meiotic prophase in mammals.
Meiosis is a double-division process that is preceded by only one DNA replication event to produce haploid gametes. The defining event in meiosis is prophase I, during which chromosome pairs locate each other, become physically connected, and exchange genetic information. Although many aspects of this process have been elucidated in lower organisms, there has been scant information available until now about the process in mammals. Recent advances in genetic analysis, especially in mice and humans, have revealed many genes that play essential roles in meiosis in mammals. These include cell cycle-regulatory proteins that couple the exit from the premeiotic DNA synthesis to the progression through prophase I, the chromosome structural proteins involved in synapsis, and the repair and recombination proteins that process the recombination events. Failure to adequately repair the DNA damage caused by recombination triggers meiotic checkpoints that result in ablation of the germ cells by apoptosis. These analyses have revealed surprising sexual dimorphism in the requirements of different gene products and a much less stringent checkpoint regulation in females. This may provide an explanation for the 10-fold increase in meiotic errors in females compared with males. This review provides a comprehensive analysis of the use of genetic manipulation, particularly in mice, but also of the analysis of mutations in humans, to elucidate the mechanisms that are required for traverse through prophase I.


Toxicity and quality of life after cytoreductive surgery plus hyperthermic intraperitoneal chemotherapy.
The purpose of our study was to determine the toxicity and quality of life for patients with peritoneal metastases after cytoreductive surgery (CS) plus hyperthermic intraperitoneal chemotherapy (HIPC). From 2001 to 2005, 35 consecutive patients with peritoneal metastases enrolled in a prospective trial approved by the University of Minnesota Institutional Review Board. Their primary cancer sites included the appendix (19 patients), colon (7), mesothelioma (3), stomach (2), small bowel (2), gallbladder (1), and unknown (1). We performed CS in an effort to remove all or nearly all peritoneal tumor nodules. Using a closed technique, we administered hyperthermic mitomycin C into the peritoneal cavity for 90 min. Before treatment and then at 4-month postoperative intervals, we used the functional assessment of cancer therapy-colon subscale (FACT-C) instrument to assess the patients' quality of life. The median hospital stay was 9 days; 12 patients were hospitalized at least 30 days or required readmission within 30 days after treatment. The postoperative mortality rate was 0%; adverse events occurred in 18 (51%) patients. As of December 2005, 20 patients were alive; 14 had died of progressive disease and 1 of an unrelated cause. The median survival time was 21.4 months. Quality of life measurements, including trial outcome index (TOI), FACT-colon, and FACT-general, returned to baseline 4 months after treatment and were significantly improved at 8 and 12 months. Despite early toxicity, CS plus HIPC may prolong the overall survival rate of patients with peritoneal metastases and improve quality of life measurements.


A comparison of droplet digital polymerase chain reaction (PCR), quantitative PCR and metabarcoding for species-specific detection in environmental DNA.
Targeted species-specific and community-wide molecular diagnostics tools are being used with increasing frequency to detect invasive or rare species. Few studies have compared the sensitivity and specificity of these approaches. In the present study environmental DNA from 90 filtered seawater and 120 biofouling samples was analyzed with quantitative PCR (qPCR), droplet digital PCR (ddPCR) and metabarcoding targeting the cytochrome c oxidase I (COI) and 18S rRNA genes for the Mediterranean fanworm Sabella spallanzanii. The qPCR analyses detected S. spallanzanii in 53% of water and 85% of biofouling samples. Using ddPCR S. spallanzanii was detected in 61% of water of water and 95% of biofouling samples. There were strong relationships between COI copy numbers determined via qPCR and ddPCR (water R2 = 0.81, p < .001, biofouling R2 = 0.68, p < .001); however, qPCR copy numbers were on average 125-fold lower than those measured using ddPCR. Using metabarcoding there was higher detection in water samples when targeting the COI (40%) compared to 18S rRNA (5.4%). The difference was less pronounced in biofouling samples (25% COI, 29% 18S rRNA). Occupancy modelling showed that although the occupancy estimate was higher for biofouling samples (ψ = 1.0), higher probabilities of detection were derived for water samples. Detection probabilities of ddPCR (1.0) and qPCR (0.93) were nearly double metabarcoding (0.57 to 0.27 marker dependent). Studies that aim to detect specific invasive or rare species in environmental samples should consider using targeted approaches until a detailed understanding of how community and matrix complexity, and primer biases affect metabarcoding data.


Rhizobia can induce nodules in white clover by "hijacking" mature cortical cells activated during lateral root development.
We examined a range of responses of root cortical cells to Rhizobium sp. inoculation to investigate why rhizobia preferentially nodulate legume roots in the zone of emerging root hairs, but generally fail to nodulate the mature root. We tested whether the inability to form nodules in the mature root is due to a lack of plant flavonoids to induce the bacterial genes required for nodulation or a failure of mature cortical cells to respond to Rhizobium spp. When rhizobia were inoculated in the zone of emerging root hairs, changes in beta-glucuronidase (GUS) expression from an auxin-responsive promoter (GH3), expression from three chalcone synthase promoters, and the accumulation of specific flavonoid compounds occurred in cortical cells prior to nodule formation. Rhizobia failed to induce these responses when inoculated in the mature root, even when co-inoculated with nod gene-inducing flavonoids. However, mature root hairs remained responsive to rhizobia and could support infection thread formation. This suggests that a deficiency in signal transduction is the reason for nodulation failure in the mature root. However, nodules could be initiated in the mature root at sites of lateral root emergence. A comparison between lateral root and nodule formation showed that similar patterns of GH3:gusA expression, chalcone synthase gene expression, and accumulation of a particular flavonoid compound occurred in the cortical cells involved in both processes. The results suggest that rhizobia can "hijack" cortical cells next to lateral root emergence sites because some of the early responses required for nodule formation have already been activated by the plant in those cells.


Mount st. Helens, washington, 1980 volcanic eruption: magmatic gas component during the first 16 days.
Eruption plumes of Mount St. Helens, Washington, showed low rates of sulfur dioxide emission, and ash leachates had low ratios of sulfur to chlorine. These data and the nonvesicularity of ash fragments are indicative of only a small eruptive magmatic component. The low amounts of soluble fluorine on the ashes pose no health problems. Violent magmatic activity is possible, and thus continued geochemical monitoring is advised.


Transcription of Alu DNA elements in blood cells of sporadic Creutzfeldt-Jakob disease (sCJD).
Alu DNA elements were long considered to be of no biological significance and thus have been only poorly defined. However, in the past Alu DNA elements with well-defined nucleotide sequences have been suspected to contribute to disease, but the role of Alu DNA element transcripts has rarely been investigated. For the first time, we determined in a real-time approach Alu DNA element transcription in buffy coat cells isolated from the blood of humans suffering from sporadic Creutzfeldt-Jakob disease (sCJD) and other neurodegenerative disorders. The reverse transcribed Alu transcripts were amplified and their cDNA sequences were aligned to genomic regions best fitted to database genomic Alu DNA element sequences deposited in the UCSC and NCBI data bases. Our cloned Alu RNA/cDNA sequences were widely distributed in the human genome and preferably belonged to the "young" Alu Y family. We also observed that some RNA/cDNA clones could be aligned to several chromosomes because of the same degree of identity and score to resident genomic Alu DNA elements. These elements, called paralogues, have purportedly been recently generated by retrotransposition. Along with cases of sCJD we also included cases of dementia and Alzheimer disease (AD). Each group revealed a divergent pattern of transcribed Alu elements. Chromosome 2 was the most preferred site in sCJD cases, besides chromosome 17; in AD cases chromosome 11 was overrepresented whereas chromosomes 2, 3 and 17 were preferred active Alu loci in controls. Chromosomes 2, 12 and 17 gave rise to Alu transcripts in dementia cases. The detection of putative Alu paralogues widely differed depending on the disease. A detailed data search revealed that some cloned Alu transcripts originated from RNA polymerase III transcription since the genomic sites of their Alu elements were found between genes. Other Alu DNA elements could be located close to or within coding regions of genes. In general, our observations suggest that identification and genomic localization of active Alu DNA elements could be further developed as a surrogate marker for differential gene expression in disease. A sufficient number of cases are necessary for statistical significance before Alu DNA elements can be considered useful to differentiate neurodegenerative diseases from controls.


Site-specific selfish genes as tools for the control and genetic engineering of natural populations.
Site-specific selfish genes exploit host functions to copy themselves into a defined target DNA sequence, and include homing endonuclease genes, group II introns and some LINE-like transposable elements. If such genes can be engineered to target new host sequences, then they can be used to manipulate natural populations, even if the number of individuals released is a small fraction of the entire population. For example, a genetic load sufficient to eradicate a population can be imposed in fewer than 20 generations, if the target is an essential host gene, the knockout is recessive and the selfish gene has an appropriate promoter. There will be selection for resistance, but several strategies are available for reducing the likelihood of it evolving. These genes may also be used to genetically engineer natural populations, by means of population-wide gene knockouts, gene replacements and genetic transformations. By targeting sex-linked loci just prior to meiosis one may skew the population sex ratio, and by changing the promoter one may limit the spread of the gene to neighbouring populations. The proposed constructs are evolutionarily stable in the face of the mutations most likely to arise during their spread, and strategies are also available for reversing the manipulations.


MHC class I-peptide interactions and TCR recognition.
The recent crystal structure determinations of MHC class I molecules with single bound peptides have allowed us to understand the guidelines that govern peptide binding in a given MHC allele. Evolution has provided for MHC class I molecules whose antigen binding clefts possess distinct physical and chemical properties as a result of the particular arrangement of variable residues lining the binding cleft. As a result, a given molecule binds a unique set of peptides, the position and identity of whose anchor residues are dictated by the features of the cleft. In addition to the interactions between the anchor residues and the cleft, the peptide is held in the groove by a highly conserved array of hydrogen bonds. A tantalizing application of our newfound understanding of peptide binding will be in the design of model peptides that either block or enhance immune response, in order to achieve effective treatments for a variety of disorders of the immune system.


Long-term survival of dialysis patients with bacterial endocarditis undergoing valvular replacement surgery in the United States.
Bacterial endocarditis in dialysis patients is associated with high mortality rates. The literature is limited on the long-term outcomes of valvular replacement surgery and the choice of prosthesis in dialysis patients with bacterial endocarditis. Dialysis patients hospitalized for bacterial endocarditis from 2004 to 2007 were studied retrospectively using data from the United States Renal Data System. Long-term survival of patients undergoing valve replacement surgery with tissue or nontissue valves was compared by use of the Kaplan-Meier method. A Cox proportional hazards model was used to identify independent predictors of mortality in patients undergoing valvular replacement surgery. During the study period, 11 156 dialysis patients were hospitalized for bacterial endocarditis and 1267 (11.4%) underwent valvular replacement surgery (tissue valve, 44.3%; nontissue valve, 55.7%). In the valve replacement cohort, 60% were men, 50% were white, 54% were 45 to 64 years of age, and 36% were diabetic. Estimated survival with tissue and nontissue valves at 0.5, 1, 2, and 3 years was 59% and 60%, 48% and 50%, 35% and 37%, and 25% and 30%, respectively (log-rank P=0.42). Staphylococcus was the predominant organism (66% of identified organisms). Independent predictors of mortality in patients undergoing valve replacement surgery included older age, diabetes mellitus as the cause of end-stage renal disease, surgery during index hospitalization, staphylococcus as the causative organism, and dysrhythmias as a comorbid condition. Valve replacement surgery is appropriate for well-selected dialysis patients with bacterial endocarditis but is associated with high mortality rates. Survival does not differ with tissue or nontissue prosthesis.


Transient expression of oatp organic anion transporter in mammalian cells: identification of candidate substrates.
The cDNA for the rat liver organic anion-transporting polypeptide "oatp" has been shown to encode transport of bromosulfophthalein (BSP) and bile salts in Xenopus oocytes (E. Jacquemin, B. Hagenbuch, B. Stieger, A. W. Wolkoff, and P. J. Meier. Proc. Natl. Acad. Sci. USA 91: 133-137, 1994). Because oatp mRNA is expressed strongly in the kidney, we sought to determine whether renal oatp might play a role in the known secretion of a large variety of organic anions by the kidney. We transiently expressed a full-length oatp cDNA, cloned in pSPORT, in HeLa cell monolayers using the recombinant vaccinia virus vtf7-3. We tested an array of organic anions as candidate substrates by determining their ability to compete with tracer BSP for transport. HeLa cell monolayers transfected with the oatp cDNA transported tracer BSP and taurocholate at rates substantially higher than monolayers transfected with a control plasmid. Thus good expression can be obtained with the vaccinia-HeLa system using a standard plasmid cloning vector. BSP transport varied as a function of the medium albumin, ionic conditions, and pH in a fashion similar to that in Xenopus oocytes. Several organic anions known to be secreted by the classic secretory pathway, including p-aminohippurate (PAH), phenol red, and indigo carmine (10 microM) failed to inhibit oatp-mediated BSP transport. Direct testing using tracers revealed no oatp-mediated transport of sulfate, urate, PAH, several eicosanoids, or unconjugated or conjugated bilirubin. On the other hand, BSP transport was inhibited by approximately 50% by 10 microM corticosterone sulfate, spironolactone, and several other steroids. We conclude that the functional properties of oatp expressed in the HeLa cell/vaccinia transient expression system are comparable to those following expression in Xenopus oocytes and that steroids are likely to represent high-affinity endogenous oatp substrates. The latter hypothesis is addressed in greater detail in a companion paper.


Elevated Prevalence of Suicide Attempts among Victims of Police Violence in the USA.
Recent evidence suggests that police victimization is widespread in the USA and psychologically impactful. We hypothesized that civilian-reported police victimization, particularly assaultive victimization (i.e., physical/sexual), would be associated with a greater prevalence of suicide attempts and suicidal ideation. Data were drawn from the Survey of Police-Public Encounters, a population-based survey of adults (N = 1615) residing in four US cities. Surveys assessed lifetime exposure to police victimization based on the World Health Organization domains of violence (i.e., physical, sexual, psychological, and neglect), using the Police Practices Inventory. Logistic regression models tested for associations between police victimization and (1) past 12-month suicide attempts and (2) past 12-month suicidal ideation, adjusted for demographic factors (i.e., gender, sexual orientation, race/ethnicity, income), crime involvement, past intimate partner and sexual victimization exposure, and lifetime mental illness. Police victimization was associated with suicide attempts but not suicidal ideation in adjusted analyses. Specifically, odds of attempts were greatly increased for respondents reporting assaultive forms of victimization, including physical victimization (odds ratio = 4.5), physical victimization with a weapon (odds ratio = 10.7), and sexual victimization (odds ratio = 10.2). Assessing for police victimization and other violence exposures may be a useful component of suicide risk screening in urban US settings. Further, community-based efforts should be made to reduce the prevalence of exposure to police victimization.


Increased intracellular targeting to airway cells using octaarginine-coated liposomes: in vitro assessment of their suitability for inhalation.
Delivery of macromolecular drugs to airway cells after inhalation can be limited by rapid clearance, in vivo degradation, and poor intracellular targeting. Liposome carriers offer an effective method of improving drug stability, but conventional liposomes have limited intracellular targeting capacity and are cleared rapidly by the lungs. Further modification is required to improve liposome-cell interaction and intracellular targeting. Therefore, we proposed conjugating three arginine-rich membrane translocating peptides, namely, HIV-TAT, Antennapedia, and octaarginine, to neutral liposomes as a biocompatible alternative to cationic lipids for intracellular delivery of macromolecules to airway cells. Conjugation did not significantly affect liposome stability, and each system was nebulized to produce aerosols of mean aerodynamic diameter < 1.5 microm. The peptides caused a significant (p < 0.05) increase in liposome-airway cell association compared to untagged liposomes and to DOTAP liposomes. Up to 30% of the peptide-conjugated liposomes added were bound and internalized (via a temperature-dependent, endocytic process) after just 2 h. The novel carriers all delivered encapsulated dextrans rapidly and efficiently to the cytoplasm of Calu-3 cells. Once internalized by the cells, the modified carriers localize for the most part in the cytoplasm with only a small amount of nuclear localization. These peptide-conjugated liposomes were significantly (p < 0.05) less toxic than DOTAP liposomes with octaarginine-coated liposomes the least toxic. These systems, particularly octaarginine-coated liposomes, offer many advantages for drug delivery to airway epithelial cells including increased stability, improved cell binding, and cell uptake with an improved toxicity profile.


Effects of enamel matrix derivative on Porphyromonas gingivalis.
Enamel matrix derivative (EMD) is used during periodontal surgery for the regeneration of periodontal tissue. It consists of the amelogenin fraction of porcine enamel matrix (AMEL) suspended in a vehicle of propylene glycol alginate (PGA). EMD-treated sites appear to heal with less inflammation. It has been suggested that antimicrobial properties of EMD might account for improved healing in vivo. The objectives of this study were: 1) to determine the antibacterial effects of EMD on the periodontal pathogen Porphyromonas gingivalis and 2) to establish the component(s) of EMD that are responsible for this effect. The antimicrobial effects were determined in vitro using the broth dilution assay. P. gingivalis at a starting inoculum of 10(9) colony forming units/ml was treated with EMD, AMEL, and PGA in Hank's balanced salt solution (HBSS) for 1, 3, and 24 hours. The CFU/ml of P. gingivalis recovered on enriched Brucella blood agar was determined at 6 and 10 days. EMD (containing AMEL and PGA) or PGA alone eliminated recoverable CFUs of P. gingivalis. Interestingly, AMEL in HBSS increased recoverable CFUs from 8.62 log CFU/ml to 8.93 log CFU/ml. Further analysis of the dose response at concentrations of 0.3, 3, and 30 mg/ml of AMEL in HBSS revealed that only 30 mg/ml (clinical concentration) increased CFUs of P. gingivalis relative to baseline (from 8.8 log CFU/ml to 9.2 log CFU/ml in 3 hours). Additionally, AMEL was compared to a protein control bovine serum albumin (BSA) to determine whether this effect was unique to AMEL. A marked increase in recoverable CFUs occurred with the AMEL (increasing from 8.8 log CFU/ml to 9.47 log CFU/ml), but not with BSA. EMD possesses antimicrobial properties that can be attributed to the propylene glycol alginate vehicle. The amelogenin fraction of porcine enamel matrix in enamel matrix derivative (i.e., AMEL) is not antibacterial for P. gingivalis and was shown to increase recoverable CFUs.


A seasonal idiopathic hepatitis syndrome in horses presented to a Midwestern veterinary teaching hospital.
To report history, clinical examination findings, clinicopathologic findings, diagnostic test results, treatment, and outcome in horses with a novel idiopathic hepatitis syndrome. 13 client-owned horses. Medical records of horses that were presented with fever and increased blood liver enzyme activity over a 16-month period were reviewed (December 1, 2020, to April 1, 2022). Collected data included signalment, history, clinical and clinicopathologic findings, diagnostic test results, treatment, clinical progression, and short-term outcome. Affected horses were presented between December and April of each of the 2 seasons investigated. The majority of horses developed cyclic fevers over the course of 3 weeks, during which time histologic evidence of hepatitis was observed. Histologic lesions included hepatic necrosis, neutrophilic to lymphohistiocytic inflammation, biliary epithelial injury, and portal fibrosis. Systemic inflammation was evidenced by increased serum amyloid A concentration and leukon changes. No horse developed signs of hepatic insufficiency, and all horses clinically recovered. Return of serum activity of GGT to within the reference range occurred within 16 weeks in most horses. Histologic lesions remained evident up to 27 weeks after initial presentation in 1 horse. Although an etiologic agent has not been identified, an apparently seasonal equine hepatitis syndrome was characterized by fever, systemic inflammation, increased liver enzyme activity, and histologic evidence of hepatitis. An infectious cause is suspected on the basis of histology and outcome.


Failure of cyclosporine to prevent in vivo T cell priming in man. Studies in allogeneic spleen transplantation.
Cyclosporine is a potent new immunosuppressive agent utilized in clinical organ transplantation. Available evidence suggest that it interferes with the secretion of interleukin-2. However, the long term efficacy of cyclosporine in preventing allograft rejection may depend on a relative sparing of suppressor cells early in the allogeneic response, allowing them to mature and effect a state of operational tolerance. If this is the case, cyclosporine must not affect antigen priming or recognition. Two patients in our center underwent allogeneic spleen transplant in conjunction with renal and pancreatic transplant. Both patients were treated with therapeutic levels of cyclosporine during the course of transplant. Neither developed any clinical signs of renal or pancreatic transplant rejection. Both patients developed graft-versus-host disease and eventually required allogeneic (donor) splenectomy. Studies performed on the splenocytes recovered from these specimens demonstrate alloantigen-specific cytotoxic T cell precursors. These studies demonstrate that although cyclosporine can prevent allograft rejection it does not necessarily prevent or ameliorate graft-versus-host disease. Furthermore, cyclosporine does not prevent in vivo T cell priming of alloantigen recognition. The primed cytotoxic precursors can be expanded in the presence of exogenous interleukin-2 to become fully active cytoxic cells.


The usefulness of elemental iron for cereal flour fortification: a SUSTAIN Task Force report. Sharing United States Technology to Aid in the Improvement of Nutrition.
Fortification of cereal flours may be a useful public health strategy to combat iron deficiency. Cereal flours that are used shortly after production (e.g., baking flour) can be fortified with soluble iron compounds, such as ferrous sulfate, whereas the majority of flours stored for longer periods is usually fortified with elemental iron powders to avoid unacceptable sensory changes. Elemental iron powders are less well absorbed than soluble iron compounds and they vary widely in their absorption depending on manufacturing method and physicochemical characteristics. Costs vary with powder type, but elemental iron powders are generally less expensive than ferrous sulfate. This review evaluates the usefulness of the different elemental iron powders based on results from in vitro studies, rat assays, human bioavailability studies, and efficacy studies monitoring iron status in human subjects. It concludes that, at the present time, only electrolytic iron powder can be recommended as an iron fortificant. Because it is only approximately half as well absorbed as ferrous sulfate, it should be added to provide double the amount of iron.


Suicide rates according to education with a particular focus on physicians in Norway 1960-2000.
Suicide rates are higher in certain educational groups. The highest rates are generally found in the medical and allied professions, but the empirical evidence for high suicide rates may be questionable. This study compares the rate of suicide among trained physicians, dentists, nurses, police officers and theologians with the rate among other university graduates and the general population according to sex, age and time period. Census data from 1960, 1970, 1980 and 1990 relating to education were linked to suicide as cause of death data from Statistics Norway, and followed up for the period 1960-2000, comprising 46 and 49 million person-years among men and women respectively. Physicians still have a higher rate compared with other graduates and the general population, both among males [43.0, 95% confidence interval (CI) 35.3-52.5] and females (26.1, 95% CI 15.1-44.9). Suicide rates increased steeply by age among physicians and other graduates, whereas for non-graduates the rate was highest in the 40-60 years age group. The suicide rate among female nurses was also elevated, whereas police officers seemed to have an intermediate suicide risk. The rate among theologians was low (7.0, 95% CI 2.9-16.9). The suicide rates in the 1990s were significantly lower than in the 1980s. The high suicide rates among physicians and elderly graduates are of concern. The reasons why graduates are more vulnerable than others when getting older and the low rate among theologians warrant further study.


Predictability of higher-order temporal structure of musical stimuli is associated with auditory evoked response.
Sound predictability resulting from repetitive patterns can be implicitly learned and often neither requires nor captures our conscious attention. Recently, predictive coding theory has been used as a framework to explain how predictable or expected stimuli evoke and gradually attenuate obligatory neural responses over time compared to those elicited by unpredictable events. However, these results were obtained using the repetition of simple auditory objects such as pairs of tones or phonemes. Here we examined whether the same principle would hold for more abstract temporal structures of sounds. If this is the case, we hypothesized that a regular repetition schedule of a set of musical patterns would reduce neural processing over the course of listening compared to stimuli with an irregular repetition schedule (and the same set of musical patterns). Electroencephalography (EEG) was recorded while participants passively listened to 6-8 min stimulus sequences in which five different four-tone patterns with temporally regular or irregular repetition were presented successively in a randomized order. N1 amplitudes in response to the first tone of each musical pattern were significantly less negative at the end of the regular sequence compared to the beginning, while such reduction was absent in the irregular sequence. These results extend previous findings by showing that N1 reflects automatic learning of the predictable higher-order structure of sound sequences, while continuous engagement of preattentive auditory processing is necessary for the unpredictable structure.