Mechanical ventilation with or without daily changes of in-line suction catheters.
The purpose of this study was to determine the safety and cost-effectiveness of not routinely changing in-line suction catheters for patients requiring mechanical ventilation. Patients were randomly assigned to receive either no routine in-line suction catheter changes (n = 258) or in-line suction catheter changes every 24 h (n = 263). The main outcome measure was the incidence of ventilator-associated pneumonia. Other outcomes evaluated included hospital mortality, acquired organ system derangements, duration of mechanical ventilation, lengths of intensive care and hospital stay, and the cost for in-line suction catheters. Ventilator-associated pneumonia was seen in 38 patients (14.7%) receiving no routine in-line suction catheter changes and in 39 patients (14.8%) receiving in-line suction catheter changes every 24 h (relative risk, 0.99; 95% CI, 0.66 to 1.50). No statistically significant differences for hospital mortality, lengths of stay, the number of acquired organ system derangements, death in patients with ventilator-associated pneumonia, or mortality directly attributed to ventilator-associated pneumonia were found between the two treatment groups. Patients receiving in-line suction catheter changes every 24 h had 1,224 catheter changes costing a total of $11,016; patients receiving no routine in-line suction catheter changes had a total of 93 catheter changes costing $837. Our findings suggest that the elimination of routine in-line suction catheter changes is safe and can reduce the costs associated with providing mechanical ventilation.


Sequential Dual Attention Network for Rain Streak Removal in a Single Image.
Various weather conditions, such as rain, haze, or snow, can degrade visual quality in images/videos, which may significantly degrade the performance of related applications. In this paper, a novel framework based on sequential dual attention deep network is proposed for removing rain streaks (deraining) in a single image, called by SSDRNet (Sequential dual attentionbased Single image DeRaining deep Network). Since the inherent correlation among rain steaks within an image should be stronger than that between the rain streaks and the background (non-rain) pixels, a two-stage learning strategy is implemented to better capture the distribution of rain streaks within a rainy image. The two-stage deep neural network primarily involves three blocks: residual dense blocks (RDBs), sequential dual attention blocks (SDABs), and multi-scale feature aggregation modules (MAMs), which are all delicately and specifically designed for rain removal. The two-stage strategy successfully learns very fine details of the rain steaks of the image and then clearly removes them. Extensive experimental results have shown that the proposed deep framework achieves the best performance on qualitative and quantitative metrics compared with state-of-the-art methods. The corresponding code and the trained model of the proposed SSDRNet have been available online at https://github.com/fityanul/SDAN-for-Rain-Removal.


An effective and better strategy for reducing body burden of radiostrontium.
In this study we have examined the effect of different calcium salts, Ca gluconate (CaG), Ca lactate (CaL), Ca carbonate (CaC) and Ca phosphate (CaP), on the clearance of radiostrontium (*Sr) administered either intraperitoneally (ip) (*Sr-ip group) or orally (*Sr-oral group) in rats. The influence of these Ca salts was examined in a group of animals administered *Sr ip, while the effect of three Ca salts (CaG, CaL and CaP) was studied in another group of rats given *Sr orally and compared with that of Ca alginate (CaA), normally advised for *Sr decorporation. Rats from both groups were subdivided into control and four experimental subgroups and were housed individually. The experimental subgroups were given the respective Ca salts (elemental Ca = 9 mg/rat/day) 2 h post 85Sr, and thereafter once daily. In the *Sr-ip group, CaG was administered ip while the other Ca salts were given orally. In the *Sr-oral group all Ca salts were administered orally. In addition, the diet of all the experimental subgroups was supplemented with the respective Ca salts to 2% elemental Ca. The whole-body retention (WBR) of *Sr in animals treated with Ca salts was found to be significantly reduced from 50-60% at 24 h to 20-30% at the end of 15 days compared with 70-80% at 24 h to 50-60% at the end of 15 days in the untreated control animals. The results strongly suggest that CaA could be replaced by any of the commonly used Ca salts for curtailing the WBR of *Sr. CaG which was administered ip, in the *Sr-ip group, was found to be more effective in reducing the WBR of *Sr.


The rationale behind intraperitoneal chemotherapy in gastrointestinal malignancies.
Local regional chemotherapy has yet to be proven superior to other methods of drug administration. While current studies are underway to assess the value of intraperitoneal chemotherapy in patients with gastrointestinal (GI) malignancies, many factors affect the success of such trials. Route of delivery and drug metabolism are all important. It is theorized that local exposure of liver with high concentrations of drug (even considering the limited efficacy of available agents) will result in improved tumor kill. In GI tumors, the putative route of spread is through the portal system to the liver. Intraperitoneal (IP) administration of drug offers an opportunity to deliver high concentrations of drug to local intraperitoneal surfaces, and, if there is sufficient hepatic extraction, IP administration also delivers a high concentration of drug to the liver. In a study of 5-fluorouracil (5-FU) in patients with metastatic colorectal carcinomas, high portal drug concentrations were achieved by this method of administration. It was concluded that IP therapy is an excellent method of delivering high concentrations of 5-FU and possibly other drugs to the hepatic parenchyma as well as to the intraperitoneal space. Since the effectiveness of such administration of still unclear, further clinical trials are indicated.


Laparoscopic approach to distal and subtotal pancreatectomy: a clockwise technique.
Laparoscopic surgery is becoming a more widely used approach for benign and malignant lesions in the neck, body, and tail of the pancreas. Recent literature reports appear to demonstrate that laparoscopic distal pancreatectomy (LDP) has clear benefits compared with open distal pancreatectomy (ODP). However, the procedure is relatively new and in some patients may remain a technically demanding operation. Twenty-nine LDPs were performed by a single surgeon during the course of 12 months by using a multistep clockwise technique described in detail below. The technique appears to simplify and standardize the approach for both the simpler and the more difficult procedure. Retrospective analysis was performed regarding perioperative outcomes. Twenty-three procedures were performed for a neoplastic process with five patients having pancreatic adenocarcinoma. There was no conversion to ODP, but one patient required a hand-assist method. Splenectomy was performed in 26 patients. Median operative time, estimated blood loss, and length of stay was 182 min, 50 ml, and 4 days respectively. Overall morbidity and pancreatic fistula rate was 17.2% and 10.3%, respectively. Median number of lymph nodes was 14, concomitant left adrenalectomy was performed in 3 patients, and margins were negative in 28 patients. LDP has been shown to be an acceptable approach to both benign and malignant disease of the distal pancreas. The technique used in this manuscript appears to facilitate a reliable and safe five-step method to perform this procedure and ensures that appropriate oncologic principles are followed through each step. Even though this is a small feasibility series focused on surgical technique, our results appear to demonstrate an acceptable pancreatic leak rate.


Role of pathogens in multiple sclerosis.
Multiple sclerosis (MS) is an inflammatory demyelinating autoimmune disease of the central nervous system (CNS). Although the etiology of MS is unknown, genetic and environmental factors play a role. Infectious pathogens are the likely environmental factors involved in the development of MS. Pathogens associated with the development or exacerbation of MS include bacteria, such as Mycoplasma pneumoniae and Chlamydia pneumoniae, the Staphylococcus aureus-produced enterotoxins that function as superantigens, viruses of the herpes virus (Epstein-Barr virus and human herpesvirus 6) and human endogenous retrovirus (HERV) families and the protozoa Acanthamoeba castellanii. Evidence, from studies with humans and animal models, supporting the association of these various pathogens with the development and/or exacerbation of MS will be discussed along with the potential mechanisms including molecular mimicry, epitope spreading and bystander activation. In contrast, infection with certain parasites such as helminthes (Schistosoma mansoni, Fasciola hepatica, Hymenolepis nana, Trichuris trichiura, Ascaris lumbricoides, Strongyloides stercolaris, Enterobius vermicularis) appears to protect against the development or exacerbation of MS. Evidence supporting the ability of parasitic infections to protect against disease will be discussed along with a brief summary of a recent Phase I clinical trial testing the ability of Trichuris suis ova treatment to improve the clinical course of MS. A complex interaction between the CNS (including the blood-brain barrier), multiple infections with various infectious agents (occurring in the periphery or within the CNS), and the immune response to those various infections may have to be deciphered before the etiology of MS can be fully understood.


Communication About Advance Directives and End-of-Life Care Options Among Internal Medicine Residents.
Despite increasing awareness about the importance of discussing end-of-life (EOL) care options with terminally ill patients and families, many physicians remain uncomfortable with these discussions. The objective of the study was to examine perceptions of and comfort with EOL care discussions among a group of internal medicine residents and the extent to which comfort with these discussions has improved over time. In 2013, internal medicine residents at a large academic medical center were asked to participate in an on-line survey that assessed their attitudes and experiences with discussing EOL care with terminally-ill patients. These results were compared to data from a similar survey residents in the same program completed in 2006. Eighty-three (50%) residents completed the 2013 survey. About half (52%) felt strongly that they were able to have open, honest discussions with patients and families, while 71% felt conflicted about whether CPR was in the patient's best interest. About half (53%) felt strongly that it was okay for them to tell a patient/family member whether or not CPR was a good idea for them. Compared to 2006 respondents, the 2013 cohort felt they had more lectures about EOL communication, and had watched an attending have an EOL discussion more often. Modest improvements were made over time in trainees' exposure to EOL discussions; however, many residents remain uncomfortable and conflicted with having EOL care discussions with their patients. More effective training approaches in EOL communication are needed to train the next generation of internists.


[Clinical and morphological features and treatment for lobular breast cancer].
The study presents data on the treatment of lobular breast cancer. On the basis of the studied morphological and molecular-biological characteristics there were substantiated types of treatment that improved the survival of patients with invasive lobular carcinoma. It was showed the efficacy of systemic treatment for lobular breast cancer and also studied long-term outcomes of lobular breast cancer depending on the morphological and biological features.


Effect of cyclic AMP phosphodiesterase inhibitors on cardiac sarcolemmal 5'-nucleotidase.
Two different membrane preparations (fraction HL and fraction S) were used to study the effect of caffeine, theophylline, dipyridamole and papaverine on cardiac sarcolemmal 5'-nucleotidase. Caffeine and theophylline inhibited the 5'-nucleotidase in both fractions, but theophylline appeared to be a more potent agent. The inhibitory effect of caffeine seemed to be the same in both membrane preparations, but the enzyme in fraction S appeared to be more sensitive to theophylline than that present in fraction HL. The inhibition was of the competitive type in both preparations. The inhibitory effect of caffeine and theophylline on 5'-nucleotidase may not be related to their actions on phosphodiesterase since dipyridamole and papaverine, which are also known to inhibit phosphodiesterase, did not affect the 5'-nucleotidase activity. It is suggested that by inhibiting 5'-nucleotidase, caffeine and theophylline may influence the production of adenosine in the myocardium and thereby may interfere with an important mechanism for the regulation of coronary blood flow.


'Hygienic' lymphocytes convey increased cancer risk.
Risk of developing inflammation-associated cancers has increased in industrialized countries during the past 30 years. One possible explanation is societal hygiene practices with use of antibiotics and Caesarian births that provide too few early life exposures of beneficial microbes. Building upon a 'hygiene hypothesis' model whereby prior microbial exposures lead to beneficial changes in CD4+ lymphocytes, here we use an adoptive cell transfer model and find that too few prior microbe exposures alternatively result in increased inflammation-associated cancer growth in susceptible recipient mice. Specifically, purified CD4+ lymphocytes collected from 'restricted flora' donors increases multiplicity and features of malignancy in intestinal polyps of recipient ApcMin/+ mice, coincident with increased inflammatory cell infiltrates and instability of the intestinal microbiota. We conclude that while a competent immune system serves to maintain intestinal homeostasis and good health, under hygienic rearing conditions CD4+ lymphocytes instead exacerbate inflammation-associated tumorigenesis, subsequently contributing to more frequent cancers in industrialized societies.


Copper absorption from human milk, cow's milk, and infant formulas using a suckling rat model.
Since copper deficiency is known to occur during infancy, it becomes important to assess copper uptake from various infant diets. We have investigated the uptake of copper from human milk, cow's milk, cow's milk formulas, cereal/milk formula and soy formula, compensating for the decay of 64Cu and using the suckling rat as a model. Radiocopper was added to the diet in trace amounts. Ultracentrifugation, ultrafiltration, and gel filtration were used to show that the added 64Cu bound to milk fractions and individual binding compounds in a manner analogous to the distribution of native copper, thus validating the use of extrinsically labeled diets. Labeled diets were intubated into 14-day-old suckling rats. Animals were killed after 6 h and tissues removed and counted. Liver copper uptake was 25% from human milk, 23% from cow's milk formula, 18% from cow's milk, 17% from premature (cow's milk based) infant formula, 17% from cereal/milk formula and 10% from soy formula. These results show that the rat pup model may provide a rapid, inexpensive, and sensitive method to assay bioavailability of copper from infant foods.


Temporal heterogeneity in blood supply in human tumor xenografts.
Temporal heterogeneities in tumor blood supply were studied by using a recently developed first-pass imaging technique. First-pass imaging movies of A-07-GFP human tumor xenografts growing in window chambers were recorded at a frame rate of approximately 9 fps and a spatial resolution of 10.8 x 10.8 microm(2) after a bolus of 155-kDa tetramethylrhodamine isothiocyanate-labeled dextran had been administered intravenously. Each tumor was subjected to imaging thrice, with 20 minutes between each repetition. Highly specific maps of the vascular network and blood supply time (BST) images (i.e., images of the time from when arterial blood enters a tumor through the main supplying artery until it reaches a vessel segment within the tumor) were produced from the movies. The tumors had one to three supplying arterioles and showed substantial temporal heterogeneity in BST. Homogeneous changes in BST in the entire vascular network were seen in tumors supplied by one arteriole. Blood supply time fluctuations in tumor subregions were observed in tumors having two or three supplying arterioles. In addition, individual vessel segments frequently showed significant changes in BST with time. High-magnification transmission microscopy imaging substantiated that BST changes could be a consequence of arterial/arteriolar vasomotor activity, vessel wall compression, varying flow rate, and vascular stasis.


Sharing patient care records over the World Wide Web.
In order to obtain appropriate medical care, patients can be referred or transported from one hospital to another based on the capacity, capability and quality of medical care provided by hospitals. Therefore, enabling patient care records to be shared among hospitals is essential not only in delivering the quality of medical care services but also in saving medical expenses. Currently, most patient care records are paper-based and not well organized. Hence, they are usually incomplete and can hardly be accessed in time. The authors in this paper present methods to structure and represent patient care records, design mechanisms for interpreting and integrating the XML-based patient care records into the existing hospital information systems. More importantly, in our approach, each significant piece of medical record is associated with a tag based on the syntax and semantics of the XML. The XML-based medical records enable a computer to capture the meaning and structure of the document on the web. The authors have developed a unified referral information system in which patient care records can be shared among hospitals over the Internet. It can not only facilitate the referral process but also maintain the integrity of a patient's medical record from distributed hospitals. The workflow of the system basically follows the existing manual system and can easily be adapted. The working group on integration of municipal hospital information systems, Department of Health, Taipei City Government, has decided to adapt this system for referral practice among the municipal hospitals.


Interaction between sleep and metabolism in Drosophila with altered octopamine signaling.
Sleep length and metabolic dysfunction are correlated, but the causal relationship between these processes is unclear. Octopamine promotes wakefulness in the fly by acting through the insulin-producing cells (IPCs) in the fly brain. To determine if insulin signaling mediates the effects of octopamine on sleep:wake behavior, we assayed flies in which insulin signaling activity was genetically altered. We found that increasing insulin signaling does not promote wake, nor does insulin appear to mediate the wake-promoting effects of octopamine. Octopamine also affects metabolism in invertebrate species, including, as we show here, Drosophila melanogaster. Triglycerides are decreased in mutants with compromised octopamine signaling and elevated in flies with increased activity of octopaminergic neurons. Interestingly, this effect is mediated at least partially by insulin, suggesting that effects of octopamine on metabolism are independent of its effects on sleep. We further investigated the relative contribution of metabolic and sleep phenotypes to the starvation response of flies with altered octopamine signaling. Hyperactivity (indicative of foraging) induced by starvation was elevated in octopamine receptor mutants, despite their high propensity for sleep, indicating that their metabolic state dictates their behavioral response under these conditions. Moreover, flies with increased octopamine signaling do not suppress sleep in response to starvation, even though they are normally hyper-aroused, most likely because of their high triglyceride levels. Together, these data suggest that observed correlations between sleep and metabolic phenotypes can result from shared molecular pathways rather than causality, and environmental conditions can lead to the dominance of one phenotype over the other.


Tim-3 is differently expressed in genetically susceptible C57BL/6 and resistant BALB/c mice during oral infection with Toxoplasma gondii.
Tim-3 has opposing roles in innate and adaptive immunities. It not only dampens CD4+ and CD8+ T cells responses but also enhances the ability of macrophages to eliminate intracellular pathogens. After peroral infection with 100 cysts of Toxoplasma gondii genetically susceptible C57BL/6 mice develop an unchecked Th1 response associated with the development of small intestinal immunopathology. Here we report that upon infection with T. gondii, both susceptible C57BL/6 and resistant BALB/c mice exhibit increased frequencies of Tim-3+ cells in spleens and mesenteric lymph nodes. The number of Tim-3+ cells was significantly higher in C57BL/6 than in BALB/c mice. Tim-3 was expressed by macrophages, dendritic, natural killer, as well as CD4+ and CD8+ T cells. Highest frequencies of Tim-3+ cells were observed at the peak of Th1 responses (day 7 post infection) concurrent with the development of ileal immunopathology. Infected Tim-3-deficient BALB/c mice did not develop ileal immunopathology nor did their parasite loads differ from those in wildtype BALB/c mice. Thus, although Tim-3 is markedly upregulated upon infection and differentially regulated in susceptible and resistant mice upon infection with T. gondii, the absence of Tim-3 is not sufficient to overcome the genetic resistance of BALB/c mice to the development of Th1-driven small intestinal immunopathology.


Elaborative encoding through self-generation enhances outcomes with errorless learning: Findings from the Skypekids memory study.
Errorless learning has demonstrated efficacy in the treatment of memory impairment in adults and older adults with acquired brain injury. In the same population, use of elaborative encoding through supported self-generation in errorless paradigms has been shown to further enhance memory performance. However, the evidence base relevant to application of both standard and self-generation forms of errorless learning in children is far weaker. We address this limitation in the present study to examine recall performance in children with brain injury (n = 16) who were taught novel age-appropriate science and social science facts through the medium of Skype. All participants were taught these facts under conditions of standard errorless learning, errorless learning with self-generation, and trial-and-error learning after which memory was tested at 5-minute, 30-minute, 1-hour and 24-hour delays. Analysis revealed no main effect of time, with participants retaining most information acquired over the 24-hour testing period, but a significant effect of condition. Notably, self-generation proved more effective than both standard errorless and trial-and-error learning. Further analysis of the data revealed that severity of attentional impairment was less detrimental to recall performance under errorless conditions. This study extends the literature to provide further evidence of the value of errorless learning methods in children with ABI and the first demonstration of the effectiveness of self-generation when delivered via the Internet.


Cloning, characterization, and sequence analysis of the clcE gene encoding the maleylacetate reductase of Pseudomonas sp. strain B13.
A 3,167-bp PstI fragment of genomic DNA from Pseudomonas sp. strain B13 was cloned and sequenced. The gene clcE consists of 1,059 nucleotides encoding a protein of 352 amino acids with a calculated mass of 37,769 Da which showed maleylacetate reductase activity. The protein had significant sequence similarities with the polypeptides encoded by tcbF of pP51 (59.4% identical positions), tfdF of pJP4 (55.1%), and tftE of Burkholderia cepacia AC1100 (53.1%). The function of TcbF as maleylacetate reductase was established by an enzyme assay.


[Acceptability of chemoprophylaxis in an endemic area for leprosy: the PEP-Hans Brazil Project].
The aim was to analyze the acceptability of chemoprophylaxis with single-dose rifampicin (PEP) in contacts, index leprosy cases, and health professionals and related factors that can influence adherence. A qualitative content analysis study was performed after application of semi-structured interviews according to the protocol proposed in the LPEP program (2016) drafted at Alta Floresta, Mato Grosso State, Brazil, in July 2016. Study participants included individuals with leprosy, contacts, and health professionals. The QRS NVivo software version 10 was used. A total of 80 individuals were contacted, including 54 (67%) contacts, 11 (14%) index cases, and 15 (19%) health professionals. 94% of the contacts (51/54) took PEP. Three PEP categories were identified: understanding, acceptance, and expectation towards the intervention. Understanding proved to be related to care by the health team. Acceptance (or lack thereof) of the medication was related to fear, trust, and protection, the strategy's operability, self-esteem, and insecurity regarding the intervention. Expectation towards the intervention was related to wellbeing, prevention of the disease, sequelae, decrease in public expenditures, and expanded access. Participants acknowledged the relevance of the PEP strategy based on the possibility of interrupting the transmission chain, reduction in new cases, and improved quality of life. Insecurity in taking the medication and the possibility of the disease manifesting itself had a negative influence on acceptance of PEP, while prior information on the PEP strategy helped strengthen trust in the health professionals and the medication's acceptance.


Durham Smith Vest-Over-Pant Technique: Simple Procedure for a Complex Problem (Post-Hypospadias Repair Fistula).
Urethrocutaneous fistula, which occurs after hypospadias surgery, is often a baffling problem and its treatment is challenging. The study aimed to evaluate the results of the simple procedure (Durham Smith vest-over-pant technique) for this complex problem (post-hypospadias repair fistula). During the period from 2011 to 2015, 20 patients with post-hypospadias repair fistulas underwent Durham Smith repair. Common age group was between 5 and 12 years. Site wise distribution of fistula was coronal 2 (10%), distal penile 7 (35%), mid-penile 7 (35%), and proximal-penile 4 (20%). Out of 20 patients, 15 had fistula of size <5 mm (75%) and 5 patients had fistula of size >5 mm (25%). All cases were repaired with Durham Smith vest-over-pant technique by a single surgeon. In case of multiple fistulas adjacent to each other, all fistulas were joined to form single fistula and repaired. We have successfully repaired all post-hypospadias surgery urethrocutaneous fistulas using the technique described by Durham Smith with 100% success rate. Durham Smith vest-over-pant technique is a simple solution for a complex problem (post hypospadias surgery penile fistulas) in properly selected patients.


The ββα fold of zinc finger proteins as a "natural" protecting group. Chemoselective synthesis of a DNA-binding zinc finger derivative.
We report the selective modification of cysteine residues engineered in peptides that have two additional cysteine residues as part of a Cys2His2 zinc finger motif. The chemoselective modification is achieved, thanks to the protecting effect exerted by the zinc cation upon coordination with the native cysteines and histidines of the zinc-finger fold. The strategy allows a straightforward synthesis of DNA binding zinc finger constructs.


Benchmark study on glyphosate-resistant cropping systems in the United States. Part 6: Timeliness of economic decision-making in implementing weed resistance management strategies.
The introduction of glyphosate-resistant (GR) crops in the late 1990s made weed control in maize, cotton and soybean simple. With the rapid adoption of GR crops, many growers began to rely solely on glyphosate for weed control. This eventually led to the evolution of GR weeds. Growers are often reluctant to adopt a weed resistance best management practice (BMP) because of the added cost of additional herbicides to weed control programs which would reduce short-term revenue. This study was designed to evaluate when a grower that is risk neutral (profit maximizing) or risk averse should adopt a weed resistance BMP. Whether a grower is risk neutral or risk averse, the optimal decision would be to adopt a weed resistance BMP when the expected loss in revenue is greater than 30% and the probability of resistance evolution is 0.1 or greater. However, if the probability of developing resistance increases to 0.3, then the best decision would be to adopt a weed resistance BMP when the expected loss is 10% or greater. Given the scenarios analyzed, risk-neutral or risk-averse growers should implement a weed resistance BMP with confidence that they have made the right decision economically and avoided the risk of lost revenue from resistance. If the grower wants to continue to see the same level of return, adoption of BMP is required.


Actinomyces marimammalium sp. nov., from marine mammals.
Three strains of a previously undescribed Actinomyces-like bacterium were isolated from samples taken from two dead seals and a porpoise. Biochemical testing and PAGE analysis of whole-cell proteins indicated the strains were phenotypically similar to each other but different from previously described Actinomyces and Arcanobacterium species. Comparative 16S rRNA gene sequencing studies showed the organisms from marine animals were genetically closely related and represent a hitherto unknown subline within the genus Actinomyces (sequence divergence values > 6% with recognized species). Based on phylogenetic and phenotypic evidence it is proposed that the unknown bacterium from the seals and a porpoise should be classified as Actinomyces marimammalium sp. nov. The type strain is CCUG 41710T.


Immunochemistry of Salmonella O-antigens: preparation of an octasaccharide-bovine serum albumin immunogen representative of Salmonella serogroup B O-antigen and characterization of the antibody response.
The O-antigenic polysaccharide of phenol-water extracted Salmonella typhimurium (O antigens 4, 12) lipopolysaccharide was enzymatically cleaved by phage P22 endorhamnosidase. An octasaccharide with the (formula: see text) structure Gal-Man-Rha-Gal-Man-Rha was isolated and shown to retain the O-antigen 4 specificity of the native polysaccharide. After oxidation of the terminal reducing rhamnose residue to the corresponding aldonic acid, the octasaccharide was covalently linked to bovine serum albumin (OLS-BSA) by use of a water-soluble carbodimide. The resulting conjugate showed O-antigen 4 specificity in enzyme-linked immunosorbent assay (ELISA) ans passive hemagglutination inhibition tests. Immunization of rabbits with the OLS-BSA conjugate gave rise to antibodies directed toward both the octasaccharide and the carrier protein. ELISA titration with synthetic disaccharide-protein conjugates as antigens revealed that the antibody titer against the mannose-rhamnose structure was higher than against the abequose-mannose structure. In rabbits immunized with heat-killed whole bacteria the titers against the two disaccharides were equal. The reason for this difference is not obvious. It is evident, however, that the OLS-BSA conjugate elicited in rabbits O-antibodies with the same specificity as whole bacteria.


Absorption, disposition, and urinary metabolism of 14C-rifabutin in rats.
14C-rifabutin was given orally (25 mg/kg) and intravenously (i.v.) (10 mg/kg) to female Sprague-Dawley rats. Radioactivity was eliminated by both the renal and fecal routes, amounting to 44.49 and 43.39% of the dose, respectively, in urine and feces at 96 h after the oral dose and to 47.81 and 40.76% of the dose, respectively, in urine and feces after the i.v. dose. Differences between the two routes of administration were negligible. Tissue distribution of radioactivity after the oral dose was investigated by the combustion technique. At 2 h, the highest concentration of radioactivity was observed in the liver, followed by the lung, abdominal adipose tissue, and spleen, whereas at 72 h, the sequence was abdominal adipose tissue, liver, spleen, bone marrow, and lung. Brain levels of radioactivity were very low. The results of whole-body autoradiography after i.v. administration confirmed the above. Whole-body autoradiography of pregnant rats showed higher concentrations of radioactivity in the uterus than in the placenta and trace levels in the fetuses up to 8 h. Radioactivity was absent in the amniotic fluid. The urinary metabolism was studied by radio-high-pressure liquid chromatography. Rifabutin accounted for 7.4 and 7.2% of the dose in 0- to 48-h urine after oral and i.v. administration, respectively. Metabolites 31-OH rifabutin and 25-O-deacetyl rifabutin amounted to 4.3 and 1.6% of the dose, respectively, after oral administration and to 2.6 and 0.7% of the dose, respectively, after i.v. administration. The remaining urinary radioactivity was mainly due to polar compounds.


[Analysis on treatment outcomes in five patients with combined en bloc liver and pancreas transplantation].
To summarize the treatment outcomes after combined en bloc liver and pancreas transplantation. Five patients with end-stage liver disease and type 2 diabetes mellitus received combined en bloc liver and pancreas transplantation after hepatectomy. Five operations were performed successfully. The operative time ranged from 9 to 16 hours and blood loss from 1600 to 3000 ml. Postoperatively, one patients developed pulmonary infection, one died of graft-versus-host disease(GVHD), and one experienced acute renal failure. No intestinal fistula, anastomotic leakage, biliary complications, chronic and acute rejection and pancreatitis were seen. Liver function index including alanine aminotransferase, aspartate aminotransferase and total bilirubin returned to normal levels a week after surgery, while levels of C peptide and blood glucose resumed within 1 to 2 weeks. Apart from 1 case died of GVHD, the other 4 maintained normal liver function during the follow up ranging from 2 to 23 months and no insulin was required for the diabetes. Combined en bloc liver and pancreas transplantation is technically feasible and an effective treatment for multi-organ diseases.


[Protective properties of Proteus ribosomal vaccine].
The results of the study of the protective properties of Proteus ribosomal vaccine prepared from P. vulgaris museum strain F-30 are presented. The vaccine showed protective effect even in a dose of 0.0001 microgram of protein per mouse; when used in immunization doses of 1 and 10 micrograms of protein per mouse, the vaccine completely protected the animals, ensuring their survival. Experiments aimed at the determination of the maximum protective effect of the vaccine were carried out. The ribosomal vaccine injected in a dose of 1 microgram of protein per mouse protected 50% of mice challenged with 10 LD50; in a dose of 10 micrograms of protein per mouse the vaccine protected 100% of the animals challenged with 3.7 and 10 LD50. Proteus ribosomal vaccine prepared from P. vulgaris museum strain was shown to render protective effect with respect to P. vulgaris and P. mirabilis clinical strains.


How bacterial pathogens eat host lipids: implications for the development of fatty acid synthesis therapeutics.
Bacterial type II fatty acid synthesis (FASII) is a target for the development of novel therapeutics. Bacteria incorporate extracellular fatty acids into membrane lipids, raising the question of whether pathogens use host fatty acids to bypass FASII and defeat FASII therapeutics. Some pathogens suppress FASII when exogenous fatty acids are present to bypass FASII therapeutics. FASII inhibition cannot be bypassed in many bacteria because essential fatty acids cannot be obtained from the host. FASII antibiotics may not be effective against all bacteria, but a broad spectrum of Gram-negative and -positive pathogens can be effectively treated with FASII inhibitors.


Association Between Peripheral Blood Oxygen Saturation (SpO2)/Fraction of Inspired Oxygen (FiO2) Ratio Time at Risk and Hospital Mortality in Mechanically Ventilated Patients.
Acute respiratory failure requiring mechanical ventilation is a leading cause of mortality in the intensive care unit. Although single peripheral blood oxygen saturation/fraction of inspired oxygen (SpO2/FiO2) ratios of hypoxemia have been evaluated to risk-stratify patients with acute respiratory distress syndrome, the utility of longitudinal SpO2/FiO2 ratios is unknown. To assess time-based SpO2/FiO2 ratios ≤ 150-SpO2/FiO2 time at risk (SF-TAR)-for predicting mortality in mechanically ventilated patients. Retrospective, observational cohort study of mechanically ventilated patients at 21 community and 2 academic hospitals. Association between the SF-TAR in the first 24 hours of ventilation and mortality was examined using multivariable logistic regression and compared with the worst recorded isolated partial pressure of arterial oxygen/fraction of inspired oxygen (P/F) ratio. In 28,758 derivation cohort admissions, every 10% increase in SF-TAR was associated with a 24% increase in adjusted odds of hospital mortality (adjusted odds ratio = 1.24; 95% confidence interval [CI] = 1.23-1.26); a similar association was observed in validation cohorts. Discrimination for mortality modestly improved with SF-TAR (area under the receiver operating characteristic curve [AUROC] = 0.81; 95% CI = 0.81-0.82) vs the worst P/F ratio (AUROC = 0.78; 95% CI = 0.78-0.79) and worst SpO2/FiO2 ratio (AUROC = 0.79; 95% CI = 0.79-0.80). The SF-TAR in the first 6 hours offered comparable discrimination for hospital mortality (AUROC = 0.80; 95% CI = 0.79-0.80) to the 24-hour SF-TAR. The SF-TAR can identify ventilated patients at increased risk of death, offering modest improvements compared with single SpO2/FiO2 and P/F ratios. This longitudinal, noninvasive, and broadly generalizable tool may have particular utility for early phenotyping and risk stratification using electronic health record data in ventilated patients.


How I do it? Endoscopic endonasal transpterygoid repair of sphenoid lateral recess cerebrospinal fluid leak after previous unsuccessful transcranial surgery.
Direct access to the sphenoid lateral recess offers the best chance of sealing spontaneous cerebrospinal fluid (CSF) rhinorrhea caused by lateral sphenoid encephaloceles of the Sternberg canal defect. We present a case of spontaneous left-sided sphenoid lateral recess CSF leak after previous unsuccessful transcranial surgery managed with an endoscopic endonasal transpterygoid approach (EETA). An anatomical-based step-by-step illustration of the EETA was presented in the surgical video. This case demonstrates the value of endoscopic endonasal transpterygoid corridor in the exposure and manipulation of the sphenoid lateral recess.


Prevalence of ECG abnormalities among adults with metabolic syndrome in a Nigerian Teaching Hospital.
Co-existence of metabolic syndrome (MetS) and electrocardiography (ECG) abnormalities heightens the risk of sudden cardiac death. However, there is a gap in evidence of how ECG changes cluster among continental Africans with or without MetS. We included 491 participants with interpretable ECG tracings who were consecutively recruited into the Cardiovascular Risk Prediction Registry (CRP). CRP is a registry of newly presenting patients into cardiology clinic of the University College Hospital, Nigeria, with a main objective of cardiovascular risk stratification to prevent cardiovascular morbidity and mortality. Using the International Diabetic Federation (IDF) criteria they were divided into those with metabolic syndrome and non-metabolic syndrome. Four hundred and ninety-one participants comprising 48.3% women with mean age 53.72±15.2 years who met the IDF criteria with complete ECG interpretations were analyzed with 44.2% (men 38.6%; women 50.2%) of the participants having MetS while 74% had ECG abnormalities. Compared to women, men had higher mean serum total cholesterol, creatinine, smoking, and alcohol use, family history of hypertension and diabetes mellitus, QT prolongation, LVH plus or minus strain pattern, and ECG abnormalities in general. Women were heavier, had higher heart rate and proportions of MetS. ECG findings among those with or without MetS were not significantly different. In men, IDF metabolic score was associated with conduction abnormalities (p=0.039) and combined ECG abnormality (p=0.042) which became more significant with an exclusion of QT prolongation (p=0.004). Also, IDF abdominal obesity was associated with QT prolongation (p=0.017), combined ECG abnormality (p=0.034) while HDLc correlated with ECG abnormalities (0.037) in men. There was no significant associations of components of metabolic syndrome with ECG abnormalities among women. There was a high prevalence of MetS and abnormal ECG among the studied population. Abnormal ECG findings were more common in men with no differential association in people with or without MetS. However, a significant association existed between certain components of MetS and ECG abnormalities in men only. Male gender and HDLc were independent predictors of ECG Abnormalities.


The problem and probability of marriage for alumnae in Progressive Era United States.
When Bryn Mawr College opened in 1885, then-president James Rhoads highlighted the precautions taken to ensure that the young women students would remain healthy, in reaction to the publicized warnings of Scottish physician Thomas S. Clouston, M.D. Dr. Clouston's concern that girls' higher education would damage their health epitomized a growing anxiety around the status of wives and mothers at a time of increased educational opportunities for the so-called 'fairer sex'. To counter these opinions, college alumnae and administrators turned to statistics. Through a combination of published statistics and informal anecdotes, this article provides an in-depth study of how marriage data were solicited, tabulated, and framed at Bryn Mawr College during the Progressive Era, contributing a detailed case-study to the historiography on the period's debates over educated women. The tension between marriage and a career in research was acutely apparent to the women in academia who were at once responsible for the statistical analysis and among the subjects under investigation. While the survey design and published results emphasized the desired outcome of marriage after graduation, these same documents also offered space for emerging professional trajectories.


Global analysis of genes differentially expressed in branching and non-branching regions of the mouse embryonic lung.
During development, the proximal and distal regions of respiratory tract undergo distinct processes that ultimately give rise to conducting airways and alveoli. To gain insights into the genetic pathways differentially activated in these regions when branching morphogenesis is initiating, we characterized their transcriptional profiles in murine rudiments isolated at embryonic (E) day 11.5. By using oligonucleotide microarrays, we identified 83 and 128 genes preferentially expressed in branching and non-branching regions, respectively. The majority of these genes (85%) had not been previously described in the lung, or in other organs. We report restricted expression patterns of 22 of these genes were by in situ hybridization. Among them in the lung potential components of the Wnt, TGF beta, FGF and retinoid pathways identified in other systems, and uncharacterized genes, such as translocases, small GTPases and splicing factors. In addition, we provide a more detailed analysis of the expression pattern and regulation of a representative gene from the distal (transforming growth factor, beta induced) and proximal (WW domain-containing protein 2) regions. Our data suggest that these genes may regulate focal developmental events specific of each of these regions during respiratory tract formation.


Increased amounts of a novel penicillin-binding protein in a strain of methicillin-resistant Staphylococcus aureus exposed to nafcillin.
In addition to the four typical penicillin-binding proteins (PBPs), a strain of heterogeneously methicillin-resistant Staphylococcus aureus produced an extra 78-kD PBP (PBP 2a) that had a low affinity for nafcillin and penicillin. Addition of nafcillin to cultures of this strain caused a rapid increase in the amount of this PBP in cell membranes. This increase occurred at subinhibitory concentrations of drug within minutes of exposure, and was blocked by inhibitors of protein and RNA synthesis. This suggests that the synthesis of PBP 2a can be stimulated by exposure to beta-lactam antibiotics. This process may, in part, explain the heterogeneity in methicillin-resistant S. aureus.


Exploring the origins of carbon in terrestrial worlds.
Given the central role of carbon in the chemistry of life, it is a fundamental question as to how carbon is supplied to the Earth, in what form and when. We provide an accounting of carbon found in solar system bodies, and in particular a comparison between the organic content of meteorites and that in identified organics in the dense interstellar medium (ISM). Based on this accounting, identified organics created by the chemistry of star formation could contain at most -15% of the organic carbon content in primitive meteorites and significantly less for cometary organics, which represent the putative contributors to starting materials for the Earth. In the ISM -30% of the elemental carbon exists as CO, either in gaseous form or in ices, with a typical abundance of -10(-4) (relative to H2). Recent observations of the TW Hya disk find that the gas phase abundance of CO is reduced by an order of magnitude compared to this value. We explore an explanation for this observation whereby the volatile CO is destroyed via gas phase processes, providing an additional source of carbon for organic material to be incorporated into planetesimals and cometesimals. This chemical processing mechanism requires warm grains (> 20 K), partially ionized gas, and sufficiently small (a(grain)) < 10 microm) grains, ie. a larger total grain surface area, such that freeze-out is efficient. Under these conditions, static (non-turbulent) chemical models predict that a large fraction of the carbon nominally sequestered in CO can be the source of carbon for a wide variety of organics that are present as ice coatings on the surfaces of warm pre-planetesimal dust grains.


Renal cell carcinoma of native kidney in Chinese renal transplant recipients: a report of 12 cases and a review of the literature.
To present and discuss the epidemiological and clinical aspects, as well as therapeutic options and outcome of de novo renal cell carcinoma (RCC) of the native kidneys in a series of Chinese renal transplant recipients. A retrospective, cohort study examining all renal transplant recipients with the diagnosis of RCC of native kidney followed up in two major regional hospitals in Hong Kong between January 2000 and December 2009. Clinical data included age, gender, cause of renal failure, symptoms at presentation, duration of transplantation, immunosuppressive therapy, and history of acquired cystic kidney disease (ACKD). Laboratory, radiographic, operative, and pathology reports were used to assess the tumor extent. Among the 1,003 renal transplant recipients recruited, 12 transplant recipients had a nephrectomy for a total of 13 RCC. The prevalence of de novo RCC was 1.3%. The mean age at diagnosis of RCC was 48.4 years, and the median time from transplantation to diagnosis was 6.1 years. ACKD was found in 6 (50%) of the patients. All patients except one were asymptomatic. pT1 disease was found in ten patients with a mean tumor size of 3.2 cm. All patients were treated successfully with radical nephrectomy. After a median follow-up of 38 months, two patients (16.7%) died. One died of sepsis, and the other died of metastatic carcinoma. With increasing data showing a better prognosis if RCC is detected early by screening, it is time to consider screening all kidney transplant recipients for ACKD and RCC.


Genomic and Metabolomic Polymorphism among Experimentally Selected Paromomycin-Resistant Leishmania donovani Strains.
Understanding the mechanism(s) underpinning drug resistance could lead to novel treatments to reverse the increased tolerance of a pathogen. In this study, paromomycin (PMM) resistance (PMMr) was induced in three Nepalese clinical strains of Leishmania donovani with different inherent susceptibilities to antimony (Sb) drugs by stepwise exposure of promastigotes to PMM. Exposure to PMM resulted in the production of mixed populations of parasites, even though a single cloned population was used at the start of selection. PMM 50% inhibitory concentration (IC50) values for PMMr parasites varied between 104 and 481 μM at the promastigote stage and 32 and 195 μM at the intracellular amastigote stage. PMM resistance was associated with increased resistance to nitric oxide at the amastigote stage but not the promastigote stage (P < 0.05). This effect was most marked in the Sb-resistant (Sbr) PMMr clone, in which PMM resistance was associated with a significant upregulation of glutathione compared to that in its wild type (P < 0.05), although there was no change in the regulation of trypanothione (detected in its oxidized form). Interestingly, PMMr strains showed an increase in either the keto acid derivative of isoleucine (Sb intermediate PMMr) or the 2-hydroxy acids derived from arginine and tyrosine (Sb susceptible PMMr and Sbr PMMr). These results are consistent with the recent finding that the upregulation of the branched-chain amino acid aminotransferase and d-lactate dehydrogenase is linked to PMMr In addition, we found that PMMr is associated with a significant increase in aneuploidy during PMM selection in all the strains, which could allow the rapid selection of genetic changes that confer a survival advantage.


Rituximab/bendamustine and rituximab/cytarabine induction therapy for transplant-eligible mantle cell lymphoma.
The addition of high-dose cytarabine to rituximab/bendamustine (RB) induction could improve outcomes for transplant-eligible patients with mantle cell lymphoma (MCL). We conducted a pooled analysis of 2 phase 2 trials and an off-trial cohort each testing 3 cycles of RB and 3 cycles of rituximab/high-dose cytarabine (RC) followed by autologous stem cell transplantation (ASCT) among untreated, transplant-eligible patients with MCL. Dana-Farber Cancer Institute (DFCI) and Washington University in St. Louis (WUSTL) led separate phase 2 trials testing sequential and alternating cycles of RB/RC, respectively. Patients treated at DFCI with sequential RB/RC off trial were retrospectively identified. Minimal residual disease (MRD) was assessed in the DFCI trial. A total of 88 patients (23 DFCI trial, 18 WUSTL trial, and 47 off trial) received RB/RC; 92% of patients completed induction, and 84% underwent planned consolidative ASCT. Grade 3 or 4 adverse events among trial patients included lymphopenia (88%), thrombocytopenia (85%), neutropenia (83%), and febrile neutropenia (15%). There were no treatment-related deaths during induction and 2 following ASCT. Among 87 response-evaluable patients, the end-of-induction overall and complete response rates were 97% and 90%, respectively. After a median follow-up of 33 months, 3-year progression-free survival and overall survival were 83% and 92%, respectively. Patients undergoing MRD testing experienced prolonged MRD negativity after ASCT with emergence of MRD occurring in only 1 patient who subsequently relapsed. RB/RC followed by ASCT achieves high rates of durable remissions in transplant-eligible patients with MCL. These trials were registered at www.clinicaltrials.gov as #NCT01661881 (DFCI trial) and #NCT02728531 (WUSTL trial).


Intravascular carbon dioxide monitoring using micro-flow colorimetry.
An intravascular carbon dioxide sensor is investigated which employs continuous perfusion of micro-quantities of reagent through silicone membrane tubing in contact with blood. Blood is sampled from a vessel by periodic withdrawal-reinfusion through a catheter and passes by the sensor membrane tubing integrated into the catheter system. Blood CO2 equilibrates across the silicone membrane causing a color change in the reagent micro-flow stream that is detected by an optical cell external to the vessel. In vivo trials on pigs demonstrate a stable sensor response, a fast response time, and high signal-to-noise ratios. The sensor also exhibits an immunity to temperature changes, reduced intravascular blood flow, photobleaching, and leaching. It has a 2 min response time, a +/-2 mmHg resolution, and minimal drift over a 12 h duration. Using a pig model, measured values compared with true values indicate a 0.998 correlation coefficient, a 1.3 mmHg precision, and a 1.7 mmHg bias.


The chondrotoxicity of single-dose corticosteroids.
Corticosteroids are commonly injected into the joint space. However, studies have not examined the chondrotoxicity of one-time injection doses. The purpose of this study is to evaluate the effect of dexamethasone sodium phosphate (Decadron), methylprednisolone acetate (Depo-Medrol), betamethasone sodium phosphate and betamethasone acetate (Celestone Soluspan), and triamcinolone acetonide (Kenalog) on human chondrocyte viability in vitro. Single-injection doses of each of the corticosteroids were separately delivered to human chondrocytes for their respective average duration of action and compared to controls using a bioreactor containing a continuous infusion pump constructed to mimic joint fluid metabolism. A 14-day time-controlled trial was also performed. A live/dead reduced biohazard viability/cytotoxicity assay was used to quantify chondrocyte viability. Over their average duration of action, betamethasone sodium phosphate/acetate solution and triamcinolone acetonide caused significant decreases in chondrocyte viability compared to control media (19.8 ± 2.9% vs. 5.2 ± 2.1%, P = 0.0025 and 10.2 ± 1.3% vs. 4.8 ± 0.9%, P = 0.0049, respectively). In the 14-day trial, only betamethasone sodium phosphate/acetate solution caused a significant decrease in chondrocyte viability compared to control media (21.5% vs. 4.6%, P < 0.001). A single-injection dose of betamethasone sodium phosphate and betamethasone acetate solution illustrated consistent and significant chondrotoxicity using a physiologically relevant in vitro model and should be used with caution. Given the observed chondrotoxicity of triamcinolone acetonide in a single trial, there may be some evidence that this medication is chondrotoxic. However, at 14 days, betamethasone sodium phosphate and betamethasone acetate was the only condition that caused significant cell death.


Excimer laser calibration system.
Excimer laser photoablation for refractive and therapeutic keratectomies has been demonstrated to be feasible and practicable. However, corneal laser ablations are not without problems, including the delivery and maintenance of a homogeneous beam. We have developed an excimer laser calibration system capable of characterizing a laser ablation profile. Beam homogeneity is determined by the analysis of a polymethylmethacrylate (PMMA)-based thin-film using video capture and image processing. The ablation profile is presented as a color-coded map. Interpolation of excimer calibration system analysis provides a three-dimensional representation of elevation profiles that correlates with two-dimensional scanning profilometry. Excimer calibration analysis was performed before treating a monkey undergoing phototherapeutic keratectomy and two human subjects undergoing myopic spherocylindrical photorefractive keratectomy. Excimer calibration analysis was performed before and after laser refurbishing. Laser ablation profiles in PMMA are resolved by the excimer calibration system to .006 microns/pulse. Correlations with ablative patterns in a monkey cornea were demonstrated with preoperative and postoperative keratometry using corneal topography, and two human subjects using video-keratography. Excimer calibration analysis predicted a central-steep-island ablative pattern with the VISX Twenty/Twenty laser, which was confirmed by corneal topography immediately postoperatively and at 1 week after reepithelialization in the monkey. Predicted central steep islands in the two human subjects were confirmed by video-keratography at 1 week and at 1 month. Subsequent technical refurbishing of the laser resulted in a beam with an overall increased ablation rate measured as microns/pulse with a donut ablation profile. A patient treated after repair of the laser electrodes demonstrated no central island. This excimer laser calibration system can precisely detect laser-beam ablation profiles. The calibration system correctly predicted central islands after excimer photoablation in a treated monkey cornea and in two treated human subjects. Detection of excimer-laser-beam ablation profiles may be useful for precise calibration of excimer lasers before human photorefractive and therapeutic surgery.


Effect of metabolic factors on the susceptibility of albino mice to experimental tuberculosis.
Mice maintained on various types of diets were found to become more susceptible to tuberculosis when deprived of food for periods of 30 hours shortly after infection. In contrast, the susceptibility of the animals to the disease was unaffected by undernutrition resulting from limitation of food intake to a low but constant daily level. The resistance of mice to tuberculosis appeared to be independent-within wide limits-of the protein content of the diet. It is true that mice fed a diet very low in protein and high in carbohydrate proved highly susceptible, but resistance was normal if part of the carbohydrate was replaced by fat (peanut oil)-without any change in the protein content of the food. Resistance to tuberculosis could be consistently and markedly decreased by adding sodium citrate (or glutarate) to a variety of diets. The survival time following infection was greatly shortened if dinitrophenol or thyroxine were administered per os in amounts sufficient to limit the weight gains of non-infected controls. There was usually a lag period of several days before the infection-enhancing effect of these metabolic stimulants became manifest. The procedures which increased the susceptibility of mice to infection with virulent tubercle bacilli also made it possible to establish in these animals a fatal infection with BCG. There was no constant relation between weight gains of uninfected mice on the various regimens, and the effect of the latter on susceptibility to tuberculosis. These findings appear compatible with, but do not prove, the hypothesis that a decrease in resistance to infection can be brought about by metabolic disturbances which cause either a depletion of the glycogen reserves of the body, or a reduction in the glycolytic activity of inflammatory cells, or an increase in the concentration of certain polycarboxylic acids and ketones in the tissues.


Volume (weight) loss and blood pressure response following thiazide diuretics.
Correlations were made between weight change (as an index of volume loss) and blood pressure (BP) reduction before and after hydrochlorothiazide treatment. A total of 343 patients with mild to moderate hypertension (95-114 mm Hg) received hydrochlorothiazide alone. The diuretic was titrated from 50 to 100 to 200 mg daily as needed until the diastolic BP fell below 90 mm Hg (goal BP) or side effects supervened. Of the 305 patients who completed the 10-week titration period, 65% attained goal BP. The effective dose of hydrochlorothiazide in 52% of these responders was 50 mg/day, and this was associated with weight loss averaging 1.58 kg. An additional 29% achieved goal BP with a similar degree of weight loss, but they required double the dose, or 100 mg/day. The remaining 19% of responders required significantly greater weight reductions averaging 3.14 kg to achieve goal BP, which necessitated hydrochlorothiazide, 200 mg/day. More blacks than whites attained goal BP despite similar degrees of weight loss in the two races. Plasma renin activity was initially higher in whites than in blacks and rose significantly more in blacks and whites requiring the greatest volume losses and the highest dose of hydrochlorothiazide to attain goal BP. Nonresponders had less weight loss than responders. Thus, diuretic dose requirements vary in different patients and are related either to different volume losses in response to a given dose or to different degrees of BP reduction in response to the same volume loss.


Induction of utrophin gene expression by heregulin in skeletal muscle cells: role of the N-box motif and GA binding protein.
The modulation of utrophin gene expression in muscle by the nerve-derived factor agrin plausibly involves the trophic factor ARIA/heregulin. Here we show that heregulin treatment of mouse and human cultured myotubes caused a approximately 2.5-fold increase in utrophin mRNA levels. Transient transfection experiments with utrophin promoter-reporter gene constructs showed that this increase resulted from an enhanced transcription of the utrophin gene. In the case of the nicotinic acetylcholine receptor delta and epsilon subunit genes, heregulin was previously reported to stimulate transcription via a conserved promoter element, the N-box, which binds the multimeric Ets-related transcription factor GA binding protein (GABP). Accordingly, site-directed mutagenesis of a single N-box motif in the utrophin gene promoter abolished the transcriptional response to heregulin. In addition, overexpression of heregulin, or of the two GABP subunits in cultured myotubes, caused an N-box-dependent increase of the utrophin promoter activity. In vivo, direct gene transfer into muscle confirmed that heregulin regulates utrophin gene expression. Finally, electrophoretic mobility shift assays and supershift experiments performed with muscle extracts revealed that the N-box of the utrophin promoter binds GABP. These findings suggest that the subsynaptic activation of transcription by heregulin via the N-box motif and GABP are conserved among genes expressed at the neuromuscular junction. Because utrophin can functionally compensate for the lack of dystrophin, the elucidation of the molecular mechanisms regulating utrophin gene transcription may ultimately lead to therapies based on utrophin expression throughout the muscle fibers of Duchenne muscular dystrophy patients.


Distal superior cerebellar artery aneurysm presenting with cerebellar infarction: report of two cases.
We report two cases of aneurysm of the distal branches of the superior cerebellar artery presenting with cerebellar infarction. In both cases, the diagnosis required close correlation of the findings from different imaging techniques, as catheter angiography failed to opacify the lesions. In one patient endovascular parent vessel occlusion was performed, whereas in the second patient the aneurysm thrombosed spontaneously. We describe the clinical and radiologic presentation of these aneurysms and discuss their pathogenesis, diagnosis, and treatment.


Accuracy of two-dimensional speckle tracking echocardiography for the detection of significant coronary stenosis.
Visual assessment of wall motion abnormalities (WMA) by 2-dimensional echocardiography (2DE) is the most semi-quantitative method used to detect coronary artery disease (CAD), but it carries many limitations. Speckle tracking echocardiography (STE) overcomes these limitations and allows an objective quantification of myocardial deformation. The aim of the study to examine the accuracy of global and segmental longitudinal strain (LS) for the detection of CAD compared with visual assessment of WMA using coronary angiography as a golden standard. The study enrolled 25 patients (mean age 51.0 ± 8.7, 64% are male) referred to coronary angiography with clinical suspicion of CAD. 2DE assessment of WMA and evaluation of LS using STE were performed using left ventricular 17-segments models. Significant CAD was defined as ≥ 50% stenosis in one or more major coronary arteries by angiography. Patients were classified into 2 groups: group I included 15 patients with significant CAD and group II included 10 patients with insignificant and/or absence of CAD. WM score was strongly correlated with the global LS in group I and II (R = 0.80, p < 0.0001 and R = 0.88, p < 0.0001 respectively). In all patients, 425 segments were analyzed. WMA was detected in 163 segments of 425 (38.3%) while abnormal LS was detected in 214 segments (50.3%). Compared with coronary angiography, the total sensitivity, specificity and accuracy for visual analysis and STE were (56%, 88.2%, and 60% vs. 68.6%, 77%, and 81.8% respectively). Segmental LS is more accurate for the detection of ischemic myocardial segment corresponding to functionally relevant coronary anatomy than visual assessment of WM.


Practical Poissonian-Gaussian noise modeling and fitting for single-image raw-data.
We present a simple and usable noise model for the raw-data of digital imaging sensors. This signal-dependent noise model, which gives the pointwise standard-deviation of the noise as a function of the expectation of the pixel raw-data output, is composed of a Poissonian part, modeling the photon sensing, and Gaussian part, for the remaining stationary disturbances in the output data. We further explicitly take into account the clipping of the data (over- and under-exposure), faithfully reproducing the nonlinear response of the sensor. We propose an algorithm for the fully automatic estimation of the model parameters given a single noisy image. Experiments with synthetic images and with real raw-data from various sensors prove the practical applicability of the method and the accuracy of the proposed model.


Mineralocorticoid receptor blockade and calcium channel blockade have different renoprotective effects on glomerular and interstitial injury in rats.
We hypothesized that combination treatment with the mineralocorticoid receptor antagonist eplerenone and the calcium channel blocker amlodipine elicits better renoprotective effects than monotherapy with either drug, via different mechanisms in Dahl salt-sensitive (DS) hypertensive rats. DS rats were fed a high-salt diet (4% NaCl) for 10 wk and were treated with vehicle (n = 12), eplerenone (50 mg x kg(-1) x day(-1), p.o., n = 12), amlodipine (3 mg x kg(-1) x day(-1), p.o., n = 12), or eplerenone plus amlodipine (n = 12) after 2 wk of salt feeding. Vehicle-treated DS rats developed proteinuria, which was attenuated by eplerenone or amlodipine. Interestingly, eplerenone attenuated the glomerulosclerosis and podocyte injury, but amlodipine did not. Conversely, treatment with amlodipine markedly improved interstitial fibrosis, while the effect of eplerenone was minimal. Combination treatment markedly improved proteinuria, glomerulosclerosis, podocyte injury, and interstitial fibrosis in DS rats. Renal hypoxia estimated by pimonidazole, vascular endothelial growth factor expression, and density of peritubular endothelial cells was exacerbated by salt feeding. Amlodipine, either as monotherapy or in combination, ameliorated the renal hypoxia, whereas eplerenone treatment had no effect. In conclusion, both eplerenone and amlodipine attenuated renal injuries in high salt-fed DS rats, but the targets for renoprotection differed between these two drugs, with eplerenone predominantly acting on glomeruli and amlodipine acting on interstitium. The combination of eplerenone and amlodipine improved renal injury more effectively than either monotherapy in high salt-fed DS rats, presumably by achieving their own renoprotective effects.


Olmec civilization, veracruz, Mexico: dating of the san lorenzo phase.
Archeological excavations at San Lorenzo Tenochtitlan, Veracruz, show that the Olmec sculptures of this zone are associated with the San Lorenzo phase, which can be placed in the Early Formative period (1500-800 B.C.) on the basis of ceramic comparisons. Five of six radiocarbon dates for the San Lorenzo phase fall within the 1200-900 B.C. span. The San Lorenzo phase therefore marks the beginning of Olmec civilization, and the sites forming the San Lorenzo Tenochtitlan group represent the oldest civilized communities known in Mexico or Central America.


Evaluation of biomonitoring data from the CDC National Exposure Report in a risk assessment context: perspectives across chemicals.
Biomonitoring data reported in the National Report on Human Exposure to Environmental Chemicals [NER; Centers for Disease Control and Prevention (2012)] provide information on the presence and concentrations of > 400 chemicals in human blood and urine. Biomonitoring Equivalents (BEs) and other risk assessment-based values now allow interpretation of these biomonitoring data in a public health risk context. We compared the measured biomarker concentrations in the NER with BEs and similar risk assessment values to provide an across-chemical risk assessment perspective on the measured levels for approximately 130 analytes in the NER. We identified available risk assessment-based biomarker screening values, including BEs and Human Biomonitoring-I (HBM-I) values from the German Human Biomonitoring Commission. Geometric mean and 95th percentile population biomarker concentrations from the NER were compared to the available screening values to generate chemical-specific hazard quotients (HQs) or cancer risk estimates. Most analytes in the NER show HQ values of < 1; however, some (including acrylamide, dioxin-like chemicals, benzene, xylene, several metals, di-2(ethylhexyl)phthalate, and some legacy organochlorine pesticides) approach or exceed HQ values of 1 or cancer risks of > 1 × 10-4 at the geometric mean or 95th percentile, suggesting exposure levels may exceed published human health benchmarks. This analysis provides for the first time a means for examining population biomonitoring data for multiple environmental chemicals in the context of the risk assessments for those chemicals. The results of these comparisons can be used to focus more detailed chemical-specific examination of the data and inform priorities for chemical risk management and research.


Meaning of wellness in caring science based on Rodgers's evolutionary concept analysis.
Wellness is a holistic, multidimensional, and process-oriented property on a continuum. It has been used interchangeably with and is undifferentiated from concepts such as health and well-being without an in-depth clarification of its theoretical foundations and a reflection on its meaning. The concept of wellness is frequently used, but its definition remains unclear. To conceptually and theoretically explore the concept of wellness to contribute to a deeper understanding in caring science. Rodgers' evolutionary concept analysis was applied to the theoretical investigation of data from publications of international origins. The focus was on antecedents, attributes, consequences, surrogate and related terms, and contextual references. A literature search was performed through a manual review of reference lists and an online search in CINAHL and PubMed via EBSCO, and in ProQuest. Abstracts were examined to identify relevant studies for further review. The inclusion criteria were peer-reviewed papers in English; papers published in scientific journals using the surrogate terms 'wellness', 'health', 'health care', and 'health care and wellness'; and papers discussing and/or defining the concept of wellness. Twenty-six studies met the inclusion criteria. Based on the findings from this concept analysis, a definition of wellness was developed: 'a holistic and multidimensional concept represented on a continuum of being well that goes beyond health'. Implications for nursing practice were correspondingly presented. Wellness is defined as a holistic and comprehensive multidimensional concept represented on a continuum of being well, that goes beyond health. It calls attention by applying the salutogenic perspective to health promotion in caring science. It is strongly related to individual lifestyle and health behaviour and is frequently used interchangeably with health and well-being without an in-depth clarification of its theoretical foundation.


Structure of Mycobacterial ergothioneine-biosynthesis C-S lyase EgtE.
L-ergothioneine is widely distributed among various microbes to regulate their physiology and pathogenicity within complex environments. One of the key steps in the ergothioneine-biosynthesis pathway, the C-S bond cleavage reaction, uses the pyridoxal 5'-phosphate dependent C-S lyase to produce the final product L-ergothioneine. Here, we present the crystallographic structure of the ergothioneine-biosynthesis C-S lyase EgtE from Mycobacterium smegmatis (MsEgtE) which represents the first published structure of ergothioneine-biosynthesis C-S lyases in bacteria, and show the effects of active site residues on the enzymatic reaction. The MsEgtE and the previously reported ergothioneine-biosynthesis C-S lyase Egt2 from Neurospora crassa (NcEgt2) fold similarly. However, discrepancies arise in terms of substrate recognition, as observed through sequence and structure comparison of MsEgtE and NcEgt2. The structural-based sequence alignment of the ergothioneine-biosynthesis C-S lyase from fungi and bacteria shows clear distinctions among the recognized substrate residues, but Arg348 is critical and an extremely conserved residue for substrate recognition. The α14 helix is exclusively found in the bacteria EgtE, which represent the most significant difference between bacteria EgtE and fungi Egt2, possibly resulting from the convergent evolution of bacteria and fungi.


[Vertical transmission of HGV and outcome of the infected babies].
To assess the risk of HGV mother-to-infant transmission and the clinical outcome of infected babies, we investigated 103 mother-infant couples and followed-up the infected children for 4-72 months. Twenty (19.4%) mothers were HGV-RNA positive and transmission occurred in ten (50%) babies; only one child acquired HGV and HCV infection. Maternal factors, such as history of intravenous drug use, HCV-RNA positivity, HIV coinfection, type of delivery and type of feeding were not related to HGV transmission. One HGV infected baby showed a mild hepatitis when he was also infected by Cytomegalovirus. Two babies cleared HGV within the first year of life. The HGV transmission rate is elevated but HGV infection seems to be benign, at least in a short-term follow-up.


Frontal Sinus Anatomy Variations in Race and Sex Using the International Frontal Sinus Anatomy Classification.
Objective: To identify frontal sinus anatomical variations grouped by patient sex, race, and presence of chronic rhinosinusitis with frontal sinus involvement (CRFS) using the International Frontal Sinus Anatomy Classification (IFAC) system. Methods: A retrospective review from 2015 to 2020 was performed of consecutive adult patients with computed tomography sinus imaging. Prevalence of frontal sinus cells using the IFAC system was recorded. Comparisons were made between patient race, sex, and CRFS groups. Results: A total of 184 patients (368 sides) were included, 90 (48.9%) of which had CRFS. The racial distribution was 50 white (27.2%), 50 black (27.2%), 45 Hispanic/Latino (24.5%), and 39 Asian (21.2%) patients. The supra agger cell was most prevalent in the white population (P = 0.009), and supraorbital ethmoid cells were more prevalent in the Asian population (P = 0.017). Patients with frontal sinus disease were more likely to have a supraorbital ethmoid cell (P = 0.024). Overall, CRFS was more prevalent in the Asian population (P = 0.013). Conclusion: Significant differences in frontal sinus anatomy and disease exist between patient race and sex. Supraorbital ethmoid cells are associated with the development of CRFS. These patterns in frontal sinus anatomy should be noted prior to frontal sinus surgery to improve surgical awareness and outcomes.


[Properties and conditions of formation of the fibronectin-collagen complex in a culture of human embryo fibroblasts].
In our previous study the macromolecular complexes mostly consisting of fibronectin and procollagen were isolated from human fibroblast culture media using immobilized antibodies against fibronectin. At present an attempt was made to elucidate at what stage the formation of the fibronectin-collagen complex occurs--either in the course of incubation of the immobilized antibodies with labelled proteins secreted by fibroblasts, or in the extracellular space while labelling fibroblasts, or intracellularly. The results obtained show that the fibronectin-collagen complex: 1) pre-exists even before incubation with the immobilized antibodies and 2) it is of intracellular origin. Thus, the considerable amount of the fibroblast-secreted fibronectin (no less than 20%) is released from the cell not in the free form but in the complex with procollagen. It was suggested that the fibronectin-collagen complex presents a stage in the formation of the insoluble extracellular matrix.


Adsorption and reduction of Cr(VI) by N, S co-doped porous carbon from sewage sludge and low-rank coal: Combining experiments and theoretical calculations.
Herein, a novel N, S co-doped porous carbon (S5C5-AC) for Cr(VI) removal was prepared by co-hydrothermal carbonization (HTC) of sewage sludge (SS) and low-rank coal (LC) combining with KOH modification. The results showed that S5C5-AC had excellent adsorption performance on Cr(VI), and lower pH value, higher initial concentration and longer contact time were beneficial for Cr(VI) adsorption. The adsorption kinetics and isotherms revealed that Cr(VI) adsorption by S5C5-AC was homogeneous and dominated by chemisorption. The adsorption isotherm showed that the maximum equilibrium adsorption capacity of S5C5-AC for Cr(VI) was 382.04 mg/g at 25 °C. Furthermore, the results showed that the main mechanisms for Cr(VI) removal were the pore filling, electrostatic interaction and reduction. Moreover, the electron transfer mechanism during the adsorption and reduction process was further explored at the molecular and electronic levels by density functional theory (DFT) and front orbital theory (FOT) simulations. The analysis of DFT and FOT indicated that the synergistic effect between S and N functional groups was exhibited during the Cr(VI) removal process. Considering the existence of synergistic effects between N and S functional groups during adsorption, the S and N content and form were modified collaboratively. Increasing the relative content of pyrrolic N may be the most effective pathway for improving removal performance. Besides that, S5C5-AC exhibited excellent adsorption capacity over a high coexisting ion concentration range and various actual water bodies and regeneration performance, which indicated that S5C5-AC had promising potential for the remediation of wastewater in industrial applications.


Physical attractiveness and the accumulation of social and human capital in adolescence and young adulthood: assets and distractions.
Beauty has a well-documented impact on labor market outcomes with both legal and policy implications. This monograph investigated whether this stratification is rooted in earlier developmental experiences. Specifically, we explored how high schools' dual roles as contexts of social relations and academic progress contributed to the long-term socioeconomic advantages of being physically attractive. Integrating theories from multiple disciplines, the conceptual model of this study contends that physically attractive youths' greater social integration and lesser social stigma help them accumulate psychosocial resources that support their academic achievement while also selecting them into social activities that distract from good grades. A mixed methods design, combining statistical analyses of the National Longitudinal Survey of Adolescent Health and qualitative analyses of a single high school, supported and expanded this model. The data revealed that the benefits of attractiveness flowed through greater social integration but were partially offset by social distractions, especially romantic/sexual partnerships and alcohol-related problems. Interview and ethnographic data further revealed that adolescents themselves understood how physical attractiveness could lead to favorable treatment by teachers and classmates while also enticing youth to emphasize socializing and dating, even when the latter took time from other activities (like studying) and marginalized some classmates. These patterns, in turn, predicted education, work, family, and mental health trajectories in young adulthood. The results of this interdisciplinary, theoretically grounded, mixed methods study suggest that adolescence may be a critical period in stratification by physical appearance and that the underlying developmental phenomena during this period are complex and often internally contradictory. The monograph concludes with discussion of theoretical and policy implications and recommendations for future developmental research.


The use of a general purpose computed tomography unit.
Two thousand five hundred and forty computed tomography (CT) examinations over a three year period were studied to determine the number of examinations of the body being performed, the use of body CT by clinicians from different specialties and the proportion of such patients with known or suspected malignant disease. Examinations were divided into head and body categories, the latter being sub-divided into groups on the basis of known neoplastic, suspected neoplastic and non-neoplastic disease. Forty-eight percent of examinations were of the body, and of these 44% were of patients with suspected neoplasia referred by general clinicians. The number of patients with known extracranial neoplasia referred by oncologists or radiotherapists was similar to the number of patients in whom there was no suspicion of malignancy. There is a significant demand for CT in the management of non-malignant conditions as well as in known or suspected malignant disease. The figures provided should be of value to those involved in establishing or updating CT units in general hospitals.


Monaural temporal integration and temporally selective listening in children and adults.
This study used two paradigms to investigate the development of temporal integration and temporally selective listening. Experiment 1 measured detection as a function of duration for a pure tone at 1625 or 6500 Hz. At both frequencies thresholds of children younger than 7 years old were higher than those for older children and adults. The pattern of temporal integration was similar across groups for the 6500-Hz signal, but younger children showed relatively more temporal integration for the 1625-Hz signal due to high thresholds for the briefest 1625-Hz signal. Experiment 2 measured detection thresholds for one or for three brief tone pips presented in a noise masker. In one set of conditions, the noise masker consisted of 100-ms steady bursts interleaved with 10-ms temporal gaps. In other conditions, the level of the central 50 ms of the 100-ms masking noise bursts was adjusted by either +6 or -6 dB. Children showed higher thresholds but similar temporal integration compared with adults. Overall, these data suggest that children are less efficient than adults in weighting the output of the monaural temporal window at 1625 but not 6500 Hz. Children are efficient in combining energy from brief temporal epochs that are separated by noise.


Antifungal activity of isothiocyanates and related compounds. I. Naturally occurring isothiocyanates and their analogues.
Data are presented concerning the antifungal activity of 11 natural isothiocyanates and 27 synthetized analogues in Aspergillus niger, Penicillium cyclopium, and Rhizopus oryzae, as well as in 13 additional saprophytic and parasitic fungi. A remarkable antifungal activity was observed in some analogues of benzylisothiocyanate and beta-phenylethylisothiocyanate. The latter-mentioned compounds have not been described previously. In the group of benzylisothiocyanates, a correlation, which was inversely proportional, was detected between ed(100) values for A. niger and R. oryzae and the corresponding molar solubilities of compounds in water. In contradistinction, no relationship was observed between antifungal activity and chemical reactivity of investigated derivatives.


Do nonmelanoma skin cancers develop from extra-cutaneous stem cells?
A hypothesis is presented that nonmelanoma skin cancers can develop from extra-cutaneous stem cells, and not exclusively from skin keratinocytes. This idea is supported by recent findings regarding the initiation of cancers in the digestive tract, and by a cancer stem cell model of a neoplasia. It is known that multipotent adult progenitor cells can trans-differentiate into very diverse cellular lineages and can be recruited to areas of profound tissue injury. In these settings, they might also initiate malignant transformation. Some epidemiological data and recent findings regarding mechanisms of wound healing indicate that skin cancers could also originate from bone marrow-derived or other extra-cutaneous stem cells in addition to local stem cells. It can therefore be speculated that the biology of keratinocyte stem cells derived from these sources differs from that of local epidermal stem cells, and consequently, these cells might be poorly controlled within their niches. Furthermore, in chronically inflamed skin, or in an immunodeficient patient, malignant transformation of extra-cutaneous stem cells is more likely to occur. There is one well-documented case of basal cell cancer which has arisen from donor cells in a kidney transplant recipient, but it remains unclear if this cancer developed directly from a donor-derived cell, or via fusion of such cells with premalignant keratinocytes. Hopefully, combining animal models of skin cancer initiation with experiments exploring the role of bone marrow-derived cells in skin healing will bring to light the exact mechanism of carcinogenesis of nonmelanoma skin cancers.


Malnutrition diagnoses in hospitalized patients: United States, 2010.
Malnutrition is common among hospitalized patients in the United States, and its coded prevalence is increasing. Malnutrition is known to be associated with increased morbidity, mortality and healthcare costs. Although national data indicate that the number of malnutrition diagnoses among hospital discharges has been steadily rising, an in-depth examination of the demographic and clinical characteristics of these patients has not been conducted. We examined data from the 2010 Healthcare Cost and Utilization Project (HCUP), the most recent nationally-representative data describing U.S. hospital discharges. Using ICD-9 codes, we constructed a composite variable indicating a diagnosis of malnutrition. Based on our definition, 3.2% of all U.S. hospital discharges in 2010 had this diagnosis. Relative to patients without a malnutrition diagnosis, those with the diagnosis were older, had longer lengths of stay and incurred higher costs. These patients were more likely to have 27 of 29 comorbidities assessed in HCUP. Finally, discharge to home care was twice as common among malnourished patients, and a discharge of death was more than 5 times as common among patients with a malnutrition diagnosis. Taken together, these nationally representative, cross-sectional data indicate that hospitalized patients discharged with a diagnosis of malnutrition are older and sicker and their inpatient care is more expensive than their counterparts without this diagnosis.


Decrease of Bcl-xL and augmentation of thymocyte apoptosis in GILZ overexpressing transgenic mice.
Glucocorticoids promote thymocyte apoptosis and modulate transcription of numerous genes. GILZ (glucocorticoid-induced leucine zipper), being one of them, is strongly up-regulated in the thymus. To elucidate its function we generated transgenic mice overexpressing it specifically in the T-cell lineage and characterized its influence on thymus function. In young adult transgenic mice CD4(+)CD8(+) thymocyte number was significantly decreased and ex vivo thymocyte apoptosis was increased. Apoptotic pathway analysis detected reduced antiapoptotic B-cell leukemia XL (Bcl-xL) expression and increased activation of caspase-8 and caspase-3. Time-course experiments showed that in wild-type (WT) thymocytes GILZ up-regulation was followed by sequential Bcl-xL decreased expression and activation of caspase-8 and of caspase-3. Moreover, GILZ delivered inside WT thymocytes by a fusion protein with the transactivator of transcription (TAT) peptide decreased Bcl-xL and promoted their apoptosis. In aged mice perturbation of thymic subset numbers was amplified over time, as demonstrated by a further decrease in CD4(+)CD8(+) cells and increases in CD4(+)CD8(-), CD4(-)CD8(-), and CD8(+)CD4(-) cell counts. These results support the hypothesis that GILZ participates in the regulation of thymocyte apoptosis by glucocorticoids.


Impaired striatal D2 receptor function leads to enhanced GABA transmission in a mouse model of DYT1 dystonia.
DYT1 dystonia is caused by a deletion in a glutamic acid residue in the C-terminus of the protein torsinA, whose function is still largely unknown. Alterations in GABAergic signaling have been involved in the pathogenesis of dystonia. We recorded GABA- and glutamate-mediated synaptic currents from a striatal slice preparation obtained from a mouse model of DYT1 dystonia. In medium spiny neurons (MSNs) from mice expressing human mutant torsinA (hMT), we observed a significantly higher frequency, but not amplitude, of GABAergic spontaneous inhibitory postsynaptic currents (sIPSCs) and miniature currents (mIPSCs), whereas glutamate-dependent spontaneous excitatory synaptic currents (sEPSCs) were normal. No alterations were found in mice overexpressing normal human torsinA (hWT). To identify the possible sources of the increased GABAergic tone, we recorded GABAergic Fast-Spiking (FS) interneurons that exert a feed-forward inhibition on MSNs. However, both sEPSC and sIPSC recorded from hMT FS interneurons were comparable to hWT and non-transgenic (NT) mice. In physiological conditions, dopamine (DA) D2 receptor act presynaptically to reduce striatal GABA release. Of note, application of the D2-like receptor agonist quinpirole failed to reduce the frequency of sIPSCs in MSNs from hMT as compared to hWT and NT mice. Likewise, the inhibitory effect of quinpirole was lost on evoked IPSCs both in MSNs and FS interneurons from hMT mice. Our findings demonstrate a disinhibition of striatal GABAergic synaptic activity, that can be at least partially attributed to a D2 DA receptor dysfunction.


Astaxanthin inhibits JAK/STAT-3 signaling to abrogate cell proliferation, invasion and angiogenesis in a hamster model of oral cancer.
Identifying agents that inhibit STAT-3, a cytosolic transcription factor involved in the activation of various genes implicated in tumour progression is a promising strategy for cancer chemoprevention. In the present study, we investigated the effect of dietary astaxanthin on JAK-2/STAT-3 signaling in the 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis model by examining the mRNA and protein expression of JAK/STAT-3 and its target genes. Quantitative RT-PCR, immunoblotting and immunohistochemical analyses revealed that astaxanthin supplementation inhibits key events in JAK/STAT signaling especially STAT-3 phosphorylation and subsequent nuclear translocation of STAT-3. Furthermore, astaxanthin downregulated the expression of STAT-3 target genes involved in cell proliferation, invasion and angiogenesis, and reduced microvascular density, thereby preventing tumour progression. Molecular docking analysis confirmed inhibitory effects of astaxanthin on STAT signaling and angiogenesis. Cell culture experiments with the endothelial cell line ECV304 substantiated the role of astaxanthin in suppressing angiogenesis. Taken together, our data provide substantial evidence that dietary astaxanthin prevents the development and progression of HBP carcinomas through the inhibition of JAK-2/STAT-3 signaling and its downstream events. Thus, astaxanthin that functions as a potent inhibitor of tumour development and progression by targeting JAK/STAT signaling may be an ideal candidate for cancer chemoprevention.


Off-target effects of CRISPRa on interleukin-6 expression.
Inactive fusion variants of the CRISPR-Cas9 system are increasingly being used as standard methodology to study transcription regulation. Their ability to readily manipulate the native genomic loci is particularly advantageous. In this work, we serendipitously uncover the key cytokine IL6 as an off-target of the activating derivative of CRISPR (CRISPRa) while studying RP11-326A19.4, a novel long-non coding RNA (lncRNA). Increasing RP11-326A19.4 expression in HEK293T cells via CRISPRa-mediated activation of its promoter region induced genome-wide transcriptional changes, including upregulation of IL6, an important cytokine. IL6 was increased in response to distinct sgRNA targeting the RP11-326A19.4 promoter region, suggesting specificity. Loss of the cognate sgRNA recognition sites failed to abolish CRISPRa mediated activation of IL6 however, pointing to off-target effects. Bioinformatic approaches did not reveal predicted off-target binding sites. Off-target activation of IL6 was sustained and involved low level activation of known IL6 regulators. Increased IL6 remained sensitive to further activation by TNFα, consistent with the existence of independent mechanisms. This study provides experimental evidence that CRISPRa has discrete, unpredictable off-targeting limitations that must be considered when using this emerging technology.


Resolution of experimental lung injury by monocyte-derived inducible nitric oxide synthase.
Although early events in the pathogenesis of acute lung injury (ALI) have been defined, little is known about the mechanisms mediating resolution. To search for determinants of resolution, we exposed wild type (WT) mice to intratracheal LPS and assessed the response at intervals to day 10, when injury had resolved. Inducible NO synthase (iNOS) was significantly upregulated in the lung at day 4 after LPS. When iNOS-/- mice were exposed to intratracheal LPS, early lung injury was attenuated; however, recovery was markedly impaired compared with WT mice. iNOS-/- mice had increased mortality and sustained increases in markers of lung injury. Adoptive transfer of WT (iNOS+/+) bone marrow-derived monocytes or direct adenoviral gene delivery of iNOS into injured iNOS-/- mice restored resolution of ALI. Irradiated bone marrow chimeras confirmed the protective effects of myeloid-derived iNOS but not of epithelial iNOS. Alveolar macrophages exhibited sustained expression of cosignaling molecule CD86 in iNOS-/- mice compared with WT mice. Ab-mediated blockade of CD86 in iNOS-/- mice improved survival and enhanced resolution of lung inflammation. Our findings show that monocyte-derived iNOS plays a pivotal role in mediating resolution of ALI by modulating lung immune responses, thus facilitating clearance of alveolar inflammation and promoting lung repair.


Adherent invasive Escherichia coli (AIEC) strain LF82, but not Candida albicans, plays a profibrogenic role in the intestine.
Intestinal fibrosis is a frequent complication of Crohn's disease. However, the factors that cause chronicity and promote fibrogenesis are not yet understood. In the present study, we evaluated the profibrotic effects of adherent-invasive Escherichia coli (AIEC) LF82 strain and Candida albicans in the gut. Colonic fibrosis was induced in C57BL/6 mice by administration of three cycles of 2.5% (w/v) dextran sulfate sodium (DSS) for 5 weeks. LF82 and C. albicans were administered orally once at the start of each week or each cycle, respectively. Expression of markers of myofibroblast activation was determined in TGF-β1-stimulated human intestinal epithelial cells (IECs). LF82 administration exacerbated fibrosis in DSS-treated mice, revealed by increased colonic collagen deposition and expression of the profibrotic genes Col1a1, Col3a1, Fn1 and Vim. This was accompanied by enhanced gene expression of proinflammatory cytokines and chemokines, as well as more recruited inflammatory cells into the intestine. LF82 also potentiated TGF-β1-stimulated epithelial-mesenchymal transition and myofibroblast activation in IECs, by further inducing gene expression of the main mesenchymal cell markers FN1 and VIM and downregulating the IEC marker OCLN. Proinflammatory cytokines were overexpressed with LF82 in TGF-β1-stimulated IECs. Conversely, C. albicans did not affect intestinal fibrosis progression in DSS-treated mice or myofibroblast activation in TGF-β1-stimulated IECs. These results demonstrate that AIEC strain LF82, but not C. albicans, may play a major profibrogenic role in the gut.