Anti-Saccharomyces cerevisiae mannan antibodies and antineutrophil cytoplasmic autoantibodies in Greek patients with inflammatory bowel disease.
The combined measurement of perinuclear antineutrophil cytoplasmic autoantibodies (pANCA) and anti-Saccharomyces cerevisiae mannan antibodies (ASCA) has recently been suggested as a valuable diagnostic approach in inflammatory bowel disease (IBD). The aim of this study was to assess the value of detecting pANCA and ASCA in the differentiation between ulcerative colitis (UC) and Crohn's disease (CD) in a Greek population with IBD. Sera were collected from 157 patients with IBD (97 with UC, 56 with CD, and four with indeterminate colitis) and 150 healthy controls. Determination of pANCA was performed by a standard indirect immunofluorescence technique on ethanol-fixed granulocytes and ASCA by an ELISA assay. In patients with UC, sensitivity, specificity, positive predictive value, and negative predictive value of the pANCA test was 67%, 84%, 93%, and 46% respectively. These values did not change significantly when the combination of positive pANCA and negative ASCA was used. ASCA test in diagnosing CD yielded a sensitivity, specificity, positive predictive value, and negative predictive value of 39%, 89%, 54%, and 81%. The combination of pANCA negative and ASCA positive increased the positive predictive value to 77% and it was associated with small bowel disease. A positive pANCA test in Greek patients has a diagnostic value in confirming a diagnosis of UC. Measurement of pANCA and ASCA together has a rather limited value in the differential diagnosis between UC and CD but may be of help in studying disease heterogeneity.


Contributions of Robotics to the Safety and Efficacy of Invasive Monitoring With Stereoelectroencephalography.
The purpose of this review is to provide a discussion of the history and utility of robotics in invasive monitoring for epilepsy surgery using stereoelectroencephalography (sEEG). The authors conducted a literature review of available sources to describe how the advent of surgical robotics has improved the efficacy and ease of performing sEEG surgery. The sEEG method integrates anatomic, electrographic, and clinical information to test hypotheses regarding the localization of the epileptogenic zone (EZ) and has been used in Europe since the 1950s. One of the primary benefits of robot-assisted sEEG implantation techniques is the ability to seamlessly transition between both orthogonal and oblique trajectory types using a single technique. Based on available information, it is our view that, when applied appropriately, robotic sEEG can have a low rate of complications and many advantages over both non-robotic sEEG implantation and traditional craniotomy-based invasive monitoring methods.


Radial Forearm Osteocutaneous Free Flap for Reconstruction of Hard Palate With Alveolar Defect.
The radial forearm free flap is beneficial for reconstruction of large palatal defect with oronasal fistula.A 51-year-old male patient who had anterior palate defect including alveolus after the radiation therapy of malignant cancer on the nasopharyngeal area undertook the radial forearm osteocutaneous free flap to close the oronasal fisula and restore the alveolar arch. The small radial bone segment was fixed in the alveolar defect and vascular anastomoses were performed with facial vessels in neck. The donor site was closed with split thickness skin graft. All suture wounds in the oral and nasal side had healed primarily with no complication within 1 month. The patient was able to swallow soluble foods in the 3 weeks postoperatively without the leakage phenomena in the nose and decreased hypernasality nature in his speech. Grafted bone union at alveolus was confirmed by follow-up computed tomography scan. There was no complication at left forearm donor site with intact musculoskeletal function.Radial forearm osteocutaneous free flap is a versatile option for its capability of reconstruction in complicated defect of soft and hard plate with alveolar defect.


[Interaction of carbonic acid reamination and oxidation processes at various functional states of tissue].
The metabolic pathways of generation and oxidation of di- and tricarboxylic acids in mitochondria differ under the resting state and during the stimulation of energy metabolism coupled with the activation of physiological functions. The contribution of amino acids to the respiration via glutamate transamination is greater during stimulation of energy metabolism than in the resting state. This may account for the increased contribution to mitochondrial respiration of succinate, the most powerful energy source among TCA substrates. The increase of succinate in the respiration is especially apparent after addition of physiological concentrations of succinate (0.3-1.0 mM). The relationship between transamination and respiration was observed during short-term experiments with mitochondria preserving their native structure. This interaction was lost in damaged mitochondria under conditions of TCA discovery.


Clinical, biologic, and histologic features of late relapses in diffuse large cell lymphoma.
After completing chemotherapy and achieving a complete remission (CR), patients with diffuse intermediate-grade lymphoma and immunoblastic lymphoma are usually considered cured if they are able to maintain that remission continuously for 24 months. Recently, we observed a number of patients with these disorders who relapsed after a continuous CR of greater than or equal to 30 months from the beginning of therapy or 24 months from completing chemotherapy. This finding led us to examine 503 consecutive cases to determine the risk of late relapse and their clinical and biologic features. We found that the overall risk of late relapse of those who attained CR was 6.8%, but several features at presentation were associated with a high risk: (1) the presence of a divergent histology; (2) a sclerosing large cell lymphoma; (3) a diagnosis based on an extranodal site with no nodal tissue available for examination. When none of these features were present, the risk of late relapse was minimal (only 3%). When any of these features was present, the risk was 14%. Most striking was the 43% late relapse rate of patients with divergent histology. All but one of the eight B-cell tumors studied at relapse showed kappa light chain restriction. Five of these eight has a low S phase at the time of relapse, suggesting chemotherapeutic selection of a clone of cells with a low proliferative potential that could have given rise to the late relapse. Nucleic acid flow cytometry and immunophenotypic studies on three tumors at initial diagnosis and after relapse failed to support the hypothesis of a second de novo lymphoma and were consistent with a true recurrence of the original tumor. The results of salvage chemotherapy in this group of late relapses showed a high CR rate (57%) but no evidence of a trend for cure in the time to treatment failure curve. In contrast to the experience with Hodgkin's disease, retreatment with the same or a similar regimen used for the original induction was not associated with durable response. Clinicians should be aware of the potential for a late relapse in cases with divergent histology and the need for new treatment strategies for such cases.


Ascorbate in the rat lens: dependence on dietary intake.
To establish a method for sample preparation to measure ascorbate in whole lenses and to investigate whether lens ascorbate concentration is dependent on dietary ascorbate intake. Four groups of 3 young Sprague-Dawley rats each were fed chow containing L-ascorbate, either 0.0, 5.7, 57.0 or 114.0 mmol/kg for a duration of 4 weeks. Thereafter, each rat was sacrificed. The lens was extracted, photographed, and lens wet weight was measured. The lens was homogenized in 1.0 ml of 0.25% metaphosphoric acid, the homogenate was centrifuged and the supernatant ultrafiltered. The filtrate was injected into an ion exchange, reversed-phase Polypore H HPLC column equipped with a 254-nm ultraviolet detector. Samples were calibrated against an L-ascorbate standard. Polynomial regression analysis was performed on the data. All lenses were devoid of cataract. A 95% confidence interval for baseline content of ascorbate without any dietary intake was estimated to be 0.16+/-0.01 micromol/g wet weight of lens. The lens ascorbate concentration increased linearly with dietary ascorbate intake with an increased rate, estimated as a 95% confidence interval of 0.33+/-0.18 (micromol ascorbate) (g lens)-1)(mol ascorbate)-1 (kg chow) with r2=0.62. Lens ascorbate concentration linearly increases with dietary ascorbate intake without cataract development in the rat. The currently presented method for sample preparation to measure the whole-lens content of ascorbate is applicable.


Contemporary blood doping-Performance, mechanism, and detection.
Blood doping is prohibited for athletes but has been a well-described practice within endurance sports throughout the years. With improved direct and indirect detection methods, the practice has allegedly moved towards micro-dosing, that is, reducing the blood doping regime amplitude. This narrative review evaluates whether blood doping, specifically recombinant human erythropoietin (rhEpo) treatment and blood transfusions are performance-enhancing, the responsible mechanism as well as detection possibilities with a special emphasis on micro-dosing. In general, studies evaluating micro-doses of blood doping are limited. However, in randomized, double-blinded, placebo-controlled trials, three studies find that infusing as little as 130 ml red blood cells or injecting 9 IU × kg bw-1 rhEpo three times per week for 4 weeks improve endurance performance ~4%-6%. The responsible mechanism for a performance-enhancing effect following rhEpo or blood transfusions appear to be increased O2 -carrying capacity, which is accompanied by an increased muscular O2 extraction and likely increased blood flow to the working muscles, enabling the ability to sustain a higher exercise intensity for a given period. Blood doping in micro-doses challenges indirect detection by the Athlete Biological Passport, albeit it can identify ~20%-60% of the individuals depending on the sample timing. However, novel biomarkers are emerging, and some may provide additive value for detection of micro blood doping such as the immature reticulocytes or the iron regulatory hormones hepcidin and erythroferrone. Future studies should attempt to validate these biomarkers for implementation in real-world anti-doping efforts and continue the biomarker discovery.


The Impact of Sexual Abuse Committed by a Child on Mental Health in Adulthood.
Numerous research studies document the negative mental health outcomes associated with the experience of childhood sexual abuse. In addition, factors such as one's relationship with the perpetrator and the severity of the abuse predict the likelihood of future mental health problems. Less attention, however, has focused on the age of the perpetrator, and recent years have seen an increased interest in children who display sexual behavior problems. College students completed measures of mental health functioning and retrospective reports of maltreatment histories. Participants were categorized as abused by an adult (n = 48), teenager (n = 39), or another child (n = 37), and non-abused (n = 219). Victims of abuse, regardless of perpetrator age, displayed higher levels of mental health problems than non-abused participants. There were no differences between the abused groups on any of the mental health outcomes; however, individuals who were abused by other children were less likely to label their experiences as abuse.


[Bilateral angle-closure glaucoma induced by trimetoprim and sulfamethoxazole combination: case report].
Sulfamethoxazole-trimethoprim (cotrimoxazole) is an antibiotic combination widely used for infections treatment and prophylaxis. These and others sulfonamides have been implicated in a rare syndrome of choroidal effusion with transient myopia and angle-closure glaucoma. Previous cases reported in literature evolved to complete resolution after drug withdrawal. In contrast, we describe a rare case in which a patient developed the syndrome while taking cotrimoxazole, but did not recover visual acuity. A 49-year-old man started Pneumocystis carini prophylaxis with cotrimoxazole; four days later, the patient presented severe ocular pain, hyperemia and chemosis. Intraocular pressure reached more than 50 mmHg in both eyes a 360 degrees choroidal effusion occurred. Medication was removed soon after the diagnosis was suspected and intraocular pressure decreased in four days. Even so total cataract and phthisis bulbi occurred in both eyes two months later. This would be the first case in the literature in which the outcome was unfavorable despite early diagnosis and withdrawal of the drug.


Regression analysis of a future state entry time distribution conditional on a past state occupation in a progressive multistate model.
We present a nonparametric method for estimating the conditional future state entry probabilities and distributions of state entry time conditional on a past state visit when data are subject to dependent censorings in a progressive multistate model where Markovianity of the system is not assumed. These estimators are constructed using the competing risk techniques with risk sets consisting of fractional observations and inverse probability of censoring weights. The fractional observations correspond to estimates of the number of persons who ultimately enter a state from which the future state in question can be reached in one step. We then address the corresponding regression problem by combining these marginal estimators with the pseudo-value approach. The performance of our regression scheme is studied using a comprehensive simulation study. An analysis of existing data on graft-versus-host disease for bone marrow transplant individuals is presented using our novel methodology. A second analysis of another well-known data set on burn patients is also included.


Monoclonal antibodies to the thyrotropin receptor: stimulating and blocking antibodies derived from the lymphocytes of patients with Graves disease.
Human monoclonal antibodies have been generated from heterohybridomas obtained by fusing mouse myeloma cells with peripheral lymphocytes from patients with active Graves disease. This report characterizes four antibodies as presumptive thyrotropin receptor antibodies because they specifically inhibit thyrotropin binding and competitively inhibit thyrotropin-induced cAMP levels in human thyroid cells. Two of these antibodies, 208F7 and 206H3, are representative of autoimmune stimulators in Graves disease sera because they stimulate thyroid function in all assays, including the mouse bioassay; their ability to inhibit thyrotropin-induced cAMP increases in thyroid cells competitively is complemented by more than additive agonism at low (10 pM) thyrotropin concentrations. These stimulating antibodies interact more potently with human thyroid ganglioside preparations than with bovine thyroid or brain gangliosides; in contrast, they are poor inhibitors of 125I-labeled thyrotropin binding to liposomes containing the glycoprotein component of the human thyrotropin receptor. Antibodies 129H8 and 122G3 appear to be representative of inhibiting or "blocking" antibodies in Graves disease sera. Thus they have no intrinsic stimulatory action in assays of thyroid function but rather inhibit thyrotropin activity in the assays tested. These two antibodies do not react with human thyroid gangliosides but are strong inhibitors of thyrotropin binding to liposomes containing the high-affinity glycoprotein component from human, bovine, and rat thyroid membranes. The data unequivocally establish the pluritopic nature of the immunoglobulins in Graves disease and relate individual components or determinants of the thyrotropin receptor structure with specific autoimmune immunoglobulins.


Quality of Communication Life in Individuals with Broca's Aphasia and Normal Individuals: A Comparative Study.
Aphasia is a neurogenic communication disorder with significant deficits in various domains of language and communication. One such type of aphasia, which impacts the quality of life significantly is Broca's aphasia, where the individual is aware of the communication difficulty. To compare the Quality of Communication Life (QoCL) between individuals with Broca's aphasia and normal individuals. The first phase of the study translated and validated QCL scale in Tamil. The second phase involved administration of the Tamil QCL scale on 12 individuals with Broca's aphasia and 12 age matched normal adults. The marked responses were analysed on a visual analogue scale independently. Three domains of QoCL were compared between the two groups using Mann-Whitney U-test. The QoCL scores across three domains were observed to be lower in individuals with Broca's aphasia, when compared to normal adults. Individuals with Broca's aphasia expressed greater challenges in socialization/activities domain of QoCL than confidence/self-concept and roles and responsibilities domains. Information obtained on self-reported QCL scale in familiar or native language will facilitate in planning client-oriented management of aphasia.


A Participatory Group Process within a Health Collaborative to Collect and Disseminate Needs Assessment Data.
Inclusion of community-based participatory research (CBPR) principles within an epidemiological study design is anticipated to promote the effective integration of knowledge derived from research data collection into the strategic planning and collaborative effort of a group focused on improving community-based health outcomes. This article describes how CBPR principles were used throughout survey design, data collection, analyses and results dissemination. A health needs assessment survey was developed as a partnership between an academic research group from the Virginia Commonwealth University (VCU), a neighborhood-level health and wellness collaborative, and community residents. Survey development, pilot testing, data collection, results dissemination, and action among the health collaborative members were carried out using CBPR principles. Feedback from participants at each stage was collected to improve the overall process of data use within the collaborative. Data from 1,064 adult participants living in the East End of Richmond, Virginia, were collected. The use of CBPR and epidemiological approaches was successful in promoting effective collaborative efforts, as indicated by 1) sustained organization-level partner participation in the development of survey items and donation of participant incentives; 2) positive feedback from resident- and organization-level participants in a preliminary data dissemination event; 3) strong resident participation at community-wide dissemination events; 4) increases in survey-related blog traffic occurring in conjunction with community-wide dissemination events; and 5) the use of process protocols and results within similar collaboratives across the city. Use of CBPR principles with epidemiological methods is a powerful tool for facilitating effective community-level strategic planning within health collaboratives.


Thyroxine action on the rat liver nuclear thyroid-hormone receptors. Binding of thyroxine to the nuclear non-histone protein and induction of mitochondrial alpha-glycerophosphate dehydrogenase activity.
The possibility that thyroxine (T4) itself exerts the hormonal effect in vivo on the rat liver nuclear receptor was studied with the aid of iopanoic acid (IOP), an inhibitor of the conversion of T4 into tri-iodothyronine (T3). After administration of 2.4 micrograms of T4/100 g body weight to hypothyroid rats for 7 days, T4 and T3 concentrations in serum and in the liver nuclear non-histone protein (NHP) were all increased to the hyperthyroid range. Hepatic mitochondrial alpha-glycerophosphate dehydrogenase (alpha-GPD) activity and DNA content increased significantly. The equilibrium association constant (Ka) of the nuclear T3 receptor was unchanged and the maximal binding capacity (Cmax.) increased 1.4-fold. Simultaneous administration of IOP (5 mg/100 g body weight) to the rats given 2.4 micrograms of T4/100 g body weight completely blocked the conversion into T3. The serum T4 was even more increased, whereas the serum T3 decreased to the hypothyroid range. Although the NHP-bound T4 was at a concentration comparable with the rats given T4 alone, no NHP-bound T3 was detected. Yet the alpha-GPD activity was elevated 2.8-fold and the DNA content increased to the same extent as observed in the rats given T4 alone. The Ka and Cmax. of the nuclear receptor were significantly decreased. After administration of 48 or 480 micrograms of T4/100 g body weight for 3 days, serum T4 and T3 were markedly increased. The NHP-bound T3 was also increased, but no NHP-bound T4 was detected. The alpha-GPD activity was markedly elevated, but the DNA content was unchanged. The Cmax. per g of liver was increased, whereas the Ka remained unchanged. Simultaneous administration of IOP to these animals could not completely block the T4 conversion. The observed hormonal effects in the absence of nuclear T3 indicate that T4 possesses the intrinsic hormonal activities on the rat liver. T4 is less potent in induction of alpha-GPD activity but as potent in increment of hepatic DNA as T3. Although the binding site for T4 is not fully characterized, it appears to be acidic NHP. T4 is an active hormone, yet is also a prohormone of T3, offering the closest analogy with testosterone.


Raspberry pomace alters cecal microbial activity and reduces secondary bile acids in rats fed a high-fat diet.
The profile of bile acids (BA) largely depends on the enzymatic activity of the microbiota, but this can be modulated by the dietary addition of biologically active compounds, e.g., polyphenols and polyunsaturated fatty acids. The aim of this study was to examine the effect of dietary raspberry pomace as a rich source of biologically active compounds on microbial activity and the BA profile in the caecum of rats fed a high-fat diet. Wistar rats were fed the standard diet AIN-93, a high-fat diet or a modified high-fat diet enriched with 7% different types of processed raspberry pomaces produced by standard grinding and fine grinding, with or without seeds. Rats fed the high-fat diet for 8 weeks showed some disorders in liver function and cecal BA, as manifested by an increased concentration of cholesterol, total BA in the liver and cholic, deoxycholic, and β-muricholic acids in the cecal digesta. In general, irrespective of the type of raspberry pomace, these dietary preparations decreased liver cholesterol, hepatic fibroblast growth factor receptor 4, peroxisome proliferator-activated receptor alpha, cecal ammonia and favorable changed BA profile in the cecum. However, among all dietary pomaces, the finely ground preparation containing seeds had the greatest beneficial effect on the caecum by modulating bacterial activity and reducing the levels of secondary BA.


Identification of mechanisms for attenuation of the FSC043 mutant of Francisella tularensis SCHU S4.
Previously, we identified a spontaneous, essentially avirulent mutant, FSC043, of the highly virulent strain SCHU S4 of Francisella tularensis subsp. tularensis. We have now characterized the phenotype of the mutant and the mechanisms of its attenuation in more detail. Genetic and proteomic analyses revealed that the pdpE gene and most of the pdpC gene were very markedly downregulated and, as previously demonstrated, that the strain expressed partially deleted and fused fupA and fupB genes. FSC043 showed minimal intracellular replication and induced no cell cytotoxicity. The mutant showed delayed phagosomal escape; at 18 h, colocalization with LAMP-1 was 80%, indicating phagosomal localization, whereas the corresponding percentages for SCHU S4 and the ΔfupA mutant were <10%. However, a small subset of the FSC043-infected cells contained up to 100 bacteria with LAMP-1 colocalization of around 30%. The unusual intracellular phenotype was similar to that of the ΔpdpC and ΔpdpC ΔpdpE mutants. Complementation of FSC043 with the intact fupA and fupB genes did not affect the phenotype, whereas complementation with the pdpC and pdpE genes restored intracellular replication and led to marked virulence. Even higher virulence was observed after complementation with both double-gene constructs. After immunization with the FSC043 strain, moderate protection against respiratory challenge with the SCHU S4 strain was observed. In summary, FSC043 showed a highly unusual intracellular phenotype, and based on our findings, we hypothesize that the mutation in the pdpC gene makes an essential contribution to the phenotype.


Timely follow-up of positive cancer screening results: A systematic review and recommendations from the PROSPR Consortium.
Timely follow-up for positive cancer screening results remains suboptimal, and the evidence base to inform decisions on optimizing the timeliness of diagnostic testing is unclear. This systematic review evaluated published studies regarding time to follow-up after a positive screening for breast, cervical, colorectal, and lung cancers. The quality of available evidence was very low or low across cancers, with potential attenuated or reversed associations from confounding by indication in most studies. Overall, evidence suggested that the risk for poorer cancer outcomes rises with longer wait times that vary within and across cancer types, which supports performing diagnostic testing as soon as feasible after the positive result, but evidence for specific time targets is limited. Within these limitations, we provide our opinion on cancer-specific recommendations for times to follow-up and how existing guidelines relate to the current evidence. Thresholds set should consider patient worry, potential for loss to follow-up with prolonged wait times, and available resources. Research is needed to better guide the timeliness of diagnostic follow-up, including considerations for patient preferences and existing barriers, while addressing methodological weaknesses. Research is also needed to identify effective interventions for reducing wait times for diagnostic testing, particularly in underserved or low-resource settings. CA Cancer J Clin 2018;68:199-216. © 2018 American Cancer Society.


Degradation of glyphosate in water by the application of surface corona discharges.
Glyphosate (GLP) is one of the most widely applied herbicides, and is found ubiquitously in the environment. The removal of glyphosate from waste water and soil is challenging and can be achieved with chemical or biological methods, which, nevertheless, suffer from different disadvantages. The application of a physical plasma for the removal of GLP in water was examined by the application of surface corona discharges in a wire-to-cylinder setup filled with argon. The plasma was ignited at the liquid surface without any additives. By applying a photometric method, GLP was detected after derivatisation with fluorenyl methoxycarbonyl chloride, whereas phosphate was determined with ammonium molybdate. A GLP degradation rate of 90.8% could be achieved within a treatment time of 30 minutes with an estimated energy efficiency of 0.32 g/kWh.


ZEITLUPE facilitates the rhythmic movements of Nicotiana attenuata flowers.
Circadian organ movements are ubiquitous in plants. These rhythmic outputs are thought to be regulated by the circadian clock and auxin signalling, but the underlying mechanisms have not been clarified. Flowers of Nicotiana attenuata change their orientation during the daytime through a 140° arc to balance the need for pollinators and the protection of their reproductive organs. This rhythmic trait is under the control of the circadian clock and results from bending and re-straightening movements of the pedicel, stems that connect flowers to the inflorescence. Using an explant system that allowed pedicel growth and curvature responses to be characterized with high spatial and temporal resolution, we demonstrated that this movement is organ autonomous and mediated by auxin. Changes in the growth curvature of the pedicel are accompanied by an auxin gradient and dorsiventral asymmetry in auxin-dependent transcriptional responses; application of auxin transport inhibitors influenced the normal movements of this organ. Silencing the expression of the circadian clock component ZEITLUPE (ZTL) arrested changes in the growth curvature of the pedicel and altered auxin signalling and responses. IAA19-like, an Aux/IAA transcriptional repressor that is circadian regulated and differentially expressed between opposite tissues of the pedicel, and therefore possibly involved in the regulation of changes in organ curvature, physically interacted with ZTL. Together, these results are consistent with a direct link between the circadian clock and the auxin signalling pathway in the regulation of this rhythmic floral movement.


Probable fatal drug interaction between intravenous fenretinide, ceftriaxone, and acetaminophen: a case report from a New Approaches to Neuroblastoma (NANT) Phase I study.
Patients with relapsed/refractory stage 4 high-risk neuroblastoma were enrolled on a phase I study (NANT2004-03) of intravenous fenretinide emulsion. Pharmacokinetic samples were collected during and after the infusion, and the levels were measured using an HPLC system. A likely case of a fatal drug interaction between fenretinide, ceftriaxone, and acetaminophen is described, including the pharmacokinetics of fenretinide, laboratory data, and post-mortem autopsy in a pediatric neuroblastoma patient treated on this study. On Day 4 of a scheduled 5-day-infusion of intravenous fenretinide, the patient developed a fever, acetaminophen was started, ceftriaxone initiated for possible bacteremia, and fenretinide level doubled from 56 to 110 μM. Over the next three days, although blood cultures remained negative, the patient's condition deteriorated rapidly. Acute liver failure was diagnosed on Day 7, and the patient expired on Day 20 of fulminant hepatic failure with associated renal, cardiac, and hemorrhagic/coagulation toxicities. Autopsy showed extensive hemorrhagic necrosis of the liver, marked bile duct proliferation, and abundant hemosiderin, consistent with cholestasis and drug toxicity. After extensive review of patient data, the clinical course, and the literature, we conclude that observed hepatic toxicity was likely due to a drug interaction between fenretinide and concomitant ceftriaxone and acetaminophen. None of the other 16 patients treated on this study experienced significant hepatic toxicity. Although the prevalence of cholestasis with ceftriaxone usage is relatively high, the potential drug interaction with these concomitant medications has not been previously reported. Concomitant use of fenretinide, ceftriaxone, and acetaminophen should be avoided.


Spinal fusion for kyphosis in achondroplasia.
Persistent thoracolumbar kyphosis in patients with achondroplasia is typically prevented with sitting modifications and bracing. When the kyphosis persists and progresses despite bracing, spinal fusion is indicated to prevent further progression and neurologic complications. Previous reports have suggested that instrumentation in such patients carries a high risk of neurologic injury. The purpose of this study was to evaluate the safety and efficacy of a two-stage procedure to control progressive kyphosis in the patient with achondroplasia. The authors treated four such patients (4-8 years old) surgically. The first stage involved an anterior spinal fusion with instrumentation and a posterior spinal fusion, and the second stage involved an additional posterior fusion. There were no neurologic complications. Correction ranged from 23.0% to 31.25%. All patients achieved a solid fusion. These results suggest that when nonoperative treatments fail, this procedure for thoracolumbar kyphosis in the achondroplastic patient can be done safely and effectively.


Major predictors of long-term clinical outcomes after coronary revascularization in patients with unprotected left main coronary disease: analysis from the MAIN-COMPARE study.
The clinical characteristics that identify high-risk subsets of patients with unprotected left main coronary artery disease undergoing percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG) have not been well established. Between January 2000 and June 2006, 2240 patients with unprotected left main coronary artery disease underwent PCI (n=1102) or CABG (n=1138). Twenty-six preprocedural parameters were evaluated by univariate and multivariate Cox regression analysis to identify independent predictors of all-cause mortality and target-vessel revascularization. Interaction tests were performed to compare heterogeneities of effects of preprocedural parameters depending on the revascularization methods. During follow-up (median of 3.1 years), 187 patients died (78 PCI and 109 CABG) and 149 patients had target-vessel revascularization (121 PCI and 28 CABG). EuroSCORE > or =6 was an independent predictor of death in both groups. Additional independent predictors were chronic renal failure and previous congestive heart failure in the PCI group and age > or =75 years, atrial fibrillation, right coronary artery disease, and left main distal bifurcation disease in the CABG group. Interaction analysis showed no heterogeneities of the effects of variables depending on the revascularization methods. Independent predictors of target-vessel revascularization were acute coronary syndrome and left main distal bifurcation disease in the PCI group and history of coronary intervention in the CABG group. The interaction between previous PCI and treatment remained after adjustment for all independent predictors of target-vessel revascularization (interaction P=0.0345). Several clinical characteristics were identified as important preprocedural predictors of long-term adverse outcomes after percutaneous or surgical revascularization in patients with unprotected left main coronary artery disease.


A recombinant influenza A virus expressing domain III of West Nile virus induces protective immune responses against influenza and West Nile virus.
West Nile virus (WNV) continues to circulate in the USA and forms a threat to the rest of the Western hemisphere. Since methods for the treatment of WNV infections are not available, there is a need for the development of safe and effective vaccines. Here, we describe the construction of a recombinant influenza virus expressing domain III of the WNV glycoprotein E (Flu-NA-DIII) and its evaluation as a WNV vaccine candidate in a mouse model. FLU-NA-DIII-vaccinated mice were protected from severe body weight loss and mortality caused by WNV infection, whereas control mice succumbed to the infection. In addition, it was shown that one subcutaneous immunization with 10(5) TCID(50) Flu-NA-DIII provided 100% protection against challenge. Adoptive transfer experiments demonstrated that protection was mediated by antibodies and CD4+T cells. Furthermore, mice vaccinated with FLU-NA-DIII developed protective influenza virus-specific antibody titers. It was concluded that this vector system might be an attractive platform for the development of bivalent WNV-influenza vaccines.


A corrosion inhibiting layer to tackle the irreversible lithium loss in lithium metal batteries.
Reactive negative electrodes like lithium (Li) suffer serious chemical and electrochemical corrosion by electrolytes during battery storage and operation, resulting in rapidly deteriorated cyclability and short lifespans of batteries. Li corrosion supposedly relates to the features of solid-electrolyte-interphase (SEI). Herein, we quantitatively monitor the Li corrosion and SEI progression (e.g., dissolution, reformation) in typical electrolytes through devised electrochemical tools and cryo-electron microscopy. The continuous Li corrosion is validated to be positively correlated with SEI dissolution. More importantly, an anti-corrosion and interface-stabilizing artificial passivation layer comprising low-solubility polymer and metal fluoride is designed. Prolonged operations of Li symmetric cells and Li | |LiFePO4 cells with reduced Li corrosion by ~74% are achieved (0.66 versus 2.5 μAh h-1). The success can further be extended to ampere-hour-scale pouch cells. This work uncovers the SEI dissolution and its correlation with Li corrosion, enabling the durable operation of Li metal batteries by reducing the Li loss.


Intakes of Specific Categories of Vegetables and Fruits Are Inversely Associated With Depressive Symptoms Among Adults.
Epidemiological evidence on the relationships between intakes of different categories of vegetables and fruits and depressive symptoms is very limited and inconsistent, especially with no evidence from the general population. This study aimed to estimate their relationships among a large general population. The cross-sectional design was based on the National Health and Nutrition Examination Survey (2007-2014) and included 16,925 adults. Dietary information was attained from two nonconsecutive 24-hr dietary recalls. Patient Health Questionnaire was applied for measuring depressive symptoms. The associations between vegetables and fruits intakes and depressive symptoms were appraised utilizing logistic regression and restricted cubic spline. Compared with the lowest category of intake, the most-adjusted odds ratios of depressive symptoms for the highest intake category of tomatoes and tomato mixtures were 0.81 (95% confidence interval [CI], 0.66-0.99), and 0.64 (95% CI, 0.48-0.85) for dark-green vegetables, 0.67 (95% CI, 0.53-0.84) for other vegetables, 0.48 (95% CI, 0.29-0.79) for berries, 0.67 (95% CI, 0.55-0.82) for total vegetables, and 0.70 (95% CI, 0.57-0.86) for total fruits, and for the medium categories of bananas and dried fruits were 0.62 (95% CI, 0.41-0.95) and 0.39 (95% CI, 0.19-0.81), respectively. After sensitivity analysis further excluding subjects with co-morbid health conditions, these findings remained significant, except for bananas. An L-shaped relationship was observed between depressive symptoms and intake of total vegetables, while the association was linear with total fruits intake. Intakes of tomatoes and tomato mixtures, dark-green vegetables, other vegetables, berries, dried fruits, total vegetables, and total fruits were inversely related to depressive symptoms among adults.


Discretionary medical reporting of potentially unfit drivers: a questionnaire-based survey in southeast Switzerland.
In Switzerland, every physician has the right to report a patient that is potentially unfit to drive to the licensing authority without violating medical confidentiality. Verified information regarding physicians' attitudes concerning this discretionary reporting and the frequency of such reports are not available. In order to answer these questions, 635 resident physicians were sent a questionnaire. The response rate was 52%. On average, the responding physicians--for all specialties--reported 0.31 patients (SD 0.64, 95% CI 0.24-0.38) in the year before the survey and 1.00 patient (SD 1.74, 95% CI 0.81-1.20) in the past 5 years. Seventy-nine percent of the responding physicians indicated knowing the current legal requirements for driving in Switzerland. In applied logistic regression analysis, only two factors correlate significantly with reporting: male sex (odds ratio 5.4) and the specialty "general medicine" (odds ratio 3.4). Ninety-seven percent of the physicians were against abolishing medical discretionary reporting and 29% were in favor of introducing mandatory reporting. The great majority of the questioned physicians supported the discretionary reporting of drivers that are potentially unfit to drive as currently practiced in Switzerland. The importance and the necessity of a regular traffic medicine-related continuing education for medical professionals are shown by the low number of reports per physician.


Stem cell microencapsulation for phenotypic control, bioprocessing, and transplantation.
Cell microencapsulation has been utilized for decades as a means to shield cells from the external environment while simultaneously permitting transport of oxygen, nutrients, and secretory molecules. In designing cell therapies, donor primary cells are often difficult to obtain and expand to appropriate numbers, rendering stem cells an attractive alternative due to their capacities for self-renewal, differentiation, and trophic factor secretion. Microencapsulation of stem cells offers several benefits, namely the creation of a defined microenvironment which can be designed to modulate stem cell phenotype, protection from hydrodynamic forces and prevention of agglomeration during expansion in suspension bioreactors, and a means to transplant cells behind a semi-permeable barrier, allowing for molecular secretion while avoiding immune reaction. This review will provide an overview of relevant microencapsulation processes and characterization in the context of maintaining stem cell potency, directing differentiation, investigating scalable production methods, and transplanting stem cells for clinically relevant disorders.


Transcriptional regulation of CYP2C19 and its role in altered enzyme activity.
Cytochrome P450 2C19 (CYP2C19) is involved in the metabolism of several drugs that are currently in clinical use. The gene encoding CYP2C19 is polymorphic with the existence of different alleles resulting in altered enzyme activity. In addition, CYP2C19 activity is also dependent on its basal expression levels determined by the transcriptional regulation. Genetic variations located in the CYP2C19 promoter region may alter the interaction of promoter with transcription factors causing variable transcription. Genetic variants may also influence the induction, inhibition of CYP2C19 and may as well affect drug-drug interactions involving CYP2C19 substrates. The role of various transcription factors and genetic variants in the promoter region of CYP2C19 regulating its expression are discussed in this review. Furthermore, induction and inhibition of CYP2C19 by various drugs in clinically meaningful drug interactions are also discussed.


Novel Insights into the Protective Properties of ACTH(4-7)PGP (Semax) Peptide at the Transcriptome Level Following Cerebral Ischaemia-Reperfusion in Rats.
Cerebral ischaemia is the most common cause of impaired brain function. Biologically active peptides represent potential drugs for reducing the damage that occurs after ischaemia. The synthetic melanocortin derivative, ACTH(4-7)PGP (Semax), has been used successfully in the treatment of patients with severe impairment of cerebral blood circulation. However, its molecular mechanisms of action within the brain are not yet fully understood. Previously, we used the transient middle cerebral artery occlusion (tMCAO) model to study the damaging effects of ischaemia-reperfusion on the brain transcriptome in rats. Here, using RNA-Seq analysis, we investigated the protective properties of the Semax peptide at the transcriptome level under tMCAO conditions. We have identified 394 differentially expressed genes (DEGs) (>1.5-fold change) in the brains of rats at 24 h after tMCAO treated with Semax relative to saline. Following tMCAO, we found that Semax suppressed the expression of genes related to inflammatory processes and activated the expression of genes related to neurotransmission. In contrast, ischaemia-reperfusion alone activated the expression of inflammation-related genes and suppressed the expression of neurotransmission-related genes. Therefore, the neuroprotective action of Semax may be associated with a compensation of mRNA expression patterns that are disrupted during ischaemia-reperfusion conditions.


Committing to quantum resistance: a slow defence for Bitcoin against a fast quantum computing attack.
Quantum computers are expected to have a dramatic impact on numerous fields due to their anticipated ability to solve classes of mathematical problems much more efficiently than their classical counterparts. This particularly applies to domains involving integer factorization and discrete logarithms, such as public key cryptography. In this paper, we consider the threats a quantum-capable adversary could impose on Bitcoin, which currently uses the Elliptic Curve Digital Signature Algorithm (ECDSA) to sign transactions. We then propose a simple but slow commit-delay-reveal protocol, which allows users to securely move their funds from old (non-quantum-resistant) outputs to those adhering to a quantum-resistant digital signature scheme. The transition protocol functions even if ECDSA has already been compromised. While our scheme requires modifications to the Bitcoin protocol, these can be implemented as a soft fork.


Does a facultative mutualism limit species range expansion?
The availability and quality of mutualists beyond a species' range edge may limit range expansion. With the legume Chamaecrista fasciculata, we asked to what extent the availability and quality of rhizobia beyond the range edge limits host range expansion. We tested the effect of rhizobia availability on plant growth by transplanting seed from three locations into five sites spanning C. fasciculata's range (interior, at the northern and western range edges, and beyond the range edges), and inoculating half the seeds with rhizobia. We recorded growth of all surviving plants, and, for the uninoculated plants, whether they had formed nodules or not. We isolated rhizobia from nodules collected on the uninoculated plants, and cross-inoculated seed from four populations (both range edge and interior populations) in the greenhouse to determine whether the quality of rhizobia differed between regions. We found that seeds transplanted beyond the range edge were less likely to be nodulated when they were not experimentally inoculated, and there was benefit to inoculation at all sites. In the greenhouse, the three inocula that formed nodules on plants, from the range interior, northern edge and beyond the northern edge, did not detectably differ in their effect on plant growth. These results suggest that low densities of suitable rhizobia beyond the range edge may limit range expansion of legume species.


Stenotrophomonas maltophilia ocular infections.
To determine if the number of ocular infections associated with Stenotrophomonas maltophilia is increasing, to identify predisposing factors, and to evaluate antimicrobial susceptibility. Retrospective review of ocular microbiology laboratory records from January 1, 1972, through December 31, 1995. Stenotrophomonas maltophilia was recovered from 15 cases of ocular infection, at a rate of one in every 1339 ocular specimens in the 1970s, one in 413 in the 1980s, and one in 363 in the 1990s through 1995. The organism was the predominant isolate in five cases and was part of a polybacterial infection in the remaining 10 cases. Eight of the 15 cases had bacterial keratitis, including one with infectious crystalline keratopathy. Of the remaining seven infections, S maltophilia was recovered from two cases of acute conjunctivitis, two infected scleral buckles (one with orbital cellulitis), two cases of infantile dacryocystitis, and one case of preseptal cellulitis. Ocular isolates of S maltophilia were resistant to the aminoglycosides and most beta-lactams, and showed variable susceptibility to the fluoroquinolones. Stenotrophomonas maltophilia is emerging as an important opportunistic ocular pathogen. Most infections by this organism occur in patients with ocular compromise, and the characteristically resistant antibiogram of S maltophilia limits the therapeutic options.


Quantification of Strain in a Porcine Model of Skin Expansion Using Multi-View Stereo and Isogeometric Kinematics.
Tissue expansion is a popular technique in plastic and reconstructive surgery that grows skin in vivo for correction of large defects such as burns and giant congenital nevi. Despite its widespread use, planning and executing an expansion protocol is challenging due to the difficulty in measuring the deformation imposed at each inflation step and over the length of the procedure. Quantifying the deformation fields is crucial, as the distribution of stretch over time determines the rate and amount of skin grown at the end of the treatment. In this manuscript, we present a method to study tissue expansion in order to gain quantitative knowledge of the deformations induced during an expansion process. This experimental protocol incorporates multi-view stereo and isogeometric kinematic analysis in a porcine model of tissue expansion. Multi-view stereo allows three-dimensional geometric reconstruction from uncalibrated sequences of images. The isogeometric kinematic analysis uses splines to describe the regional deformations between smooth surfaces with few mesh points. Our protocol has the potential to bridge the gap between basic scientific inquiry regarding the mechanics of skin expansion and the clinical setting. Eventually, we expect that the knowledge gained with our methodology will enable treatment planning using computational simulations of skin deformation in a personalized manner.


Patterns of Amyotrophic Lateral Sclerosis and Parkinsonism-Dementia on Guam.
The striking concentration of amyotrophic lateral sclerosis (ALS) and parkinsonismdementia (PD) among the Chamorro people of Guam has attracted attention since the early 1950's. Extensive studies conducted over the past 20 years have not revealed the causes of these diseases and the reasons for their remarkable concentrations on Guam. Approximately 1 in 10 Guamanian deaths over age 25 is from ALS, and another 1 in 10 is from PD. ALS still occurs approximately 50 times more frequently on Guam than in the continental United States. Advances in virologic and biochemical techniques, and long years of study, appear to be leading us to a point where some of the problems will be resolved. We shall attempt to review the pertinent background information and summarize the data and observations up to the present time.


Nurses: extending care through telehealth.
Nurses have an immense impact on the growth and scope of Telehealth as they embrace diverse roles. TeleNursing introduces transformational change which positively impacts both clients and providers, increasing access to care and reducing the time and costs associated with traveling for health care. Integration of clinical support and remote diagnostic tools inspire new uses of Telehealth, thus enabling care previously only possible in person to be delivered virtually. Nurses currently leverage Telehealth to deliver care and education, monitor clients remotely and support medical consultations. Over 90% of Island Health nurses surveyed recommend Telehealth as a care modality to clients and 100% support Telehealth as a means to increase care to vulnerable communities. Programs wish to increase uptake of TeleNursing but face numerous challenges regarding funding, resourcing, scheduling and geographical ownership. TeleNursing goes beyond clinical support and has the potential to exponentially expand Telehealth services, normalizing Telehealth as a care modality. Nurses look to Telehealth to improve their ability to partner with clients over distance, providing surgical care, maternal/pediatric care and group education.


Protein-losing enteropathy in dogs is associated with decreased fecal proteolytic activity.
Measurement of proteolytic activity in feces is a traditional method for the diagnosis of exocrine pancreatic insufficiency (EPI). A drawback of this method is the occurrence of falsely low results that may lead to a false-positive diagnosis of EPI. We hypothesized that intestinal loss of serum proteinase inhibitors in protein-losing enteropathy (PLE) may inhibit fecal proteolytic activity and be a potential source of false low results. The objective of this study was to determine the effect of PLE on fecal proteolytic activity in dogs. Fecal proteolytic activity was measured using a radial diffusion casein digestion assay in 12 samples from 4 clinically healthy control dogs and 30 samples from 16 dogs with PLE. Gastrointestinal protein loss was assessed using an ELISA to determine fecal canine alpha 1-proteinase inhibitor concentration. The relationship between the concentration of canine alpha 1-proteinase inhibitor in the feces and the diameter cleared in the casein digestion assay was determined. The mean clearing diameter was compared between control dogs and dogs with PLE. A significant negative correlation was observed between fecal canine alpha1-proteinase inhibitor concentration and casein clearing diameter (P <.001, Pearson r =.6317, r2 =.3999). Mean clearing diameter was significantly lower in dogs with PLE than in control dogs (12.63 vs 16.83 mm, P <.001, two-tailed Student's t-test). Increased fecal loss of alpha1-proteinase inhibitor in dogs with PLE is associated with a significant decrease in fecal proteolytic activity and may result in a false positive diagnosis of EPI.


[Various mechanisms of circulatory failure in septic shock].
The main parameters of hemodynamic and myocardial contractile activity, transcapillary fluid and proteins exchange have been studied in healthy and infected anaesthetized dogs with source of purulent infection during septic shock development. Besides, blood gas tension, pH and toxicity of blood and lymph are determined as well. It is shown that damage of microcirculation promoting tissue hypoxia and endogenous intoxication, is responsible for the initiation of decompensating disturbance of circulation at pus process generalization.


Subcutaneous emphysema to the head and neck resulting from a dental crown preparation.
A 59-year-old African-American female presents with subcutaneous emphysema to thehead and neck region resulting from routine dental treatment with a high speed air-driven handpiece. The patient had a chief complaint of generalized edema, dysphagia and pain to her face and neck. The patient subsequently was admitted to the intensive care unit for airway monitoring. To alert the dental community about the incidence of iatrogenic subcutaneous emphysema from a routine dental procedure, and how to recognize and manage its occurrence,


Thaw-Induced Gelation of Alginate Hydrogels for Versatile Delivery of Therapeutics.
Alginate hydrogels have been extensively used and successfully validated as delivery vehicles of bioactive factors in many tissue engineering applications. This work describes and characterizes a singular alternative method to create alginate hydrogels designated as thaw-induced gelation (TIG). The TIG method involves gelation through the time-dependent release of the polymer or crosslinker by melting into solution. Alginate TIG hydrogels were validated for spatial-temporal control delivery of different cargos including albumin, dextran, and doxorubicin. Chitosan was incorporated into TIG hydrogels to investigate the electrostatic interactions between alginate and the tested cargos. Interestingly, while 90% of doxorubicin was released after 8 h from hydrogels formed with frozen calcium, hydrogels formulated from frozen alginate took 72 h. In addition, the storage modulus of TIG hydrogels prepared from frozen alginate was double that of a hydrogel formed without freezing alginate. Therefore, the utility of TIG strategies are particularly promising for the delivery of therapeutic cargos smaller than the mesh size of the alginate hydrogel, as it enables controlled release of these cargos without any further chemical modifications of the hydrogels. These TIG alginate hydrogels with tunable mechanical properties and control over the delivery of smaller cargos could be useful in many tissue engineering applications.


Experimental verification of a predicted, hitherto unseen separation selectivity pattern in the nonaqueous capillary electrophoretic separation of weak base enantiomers by octakis (2,3-diacetyl-6-sulfato)-gamma-cyclodextrin.
The capillary electrophoretic separation of cationic enantiomers with single-isomer multivalent anionic resolving agents was reexamined with the help of the charged resolving agent migration model. Three general model parameters were identified that influence the shape of the separation selectivity and enantiomer mobility difference curves: parameter b, the binding selectivity (K(RCD)/K(SCD)), parameter s, the size selectivity (mu0(RCD)/mu0(SCD)), and parameter a, the complexation-induced alteration of the analyte's mobility (mu0(RCD)/mu0). In addition to the previously observed discontinuity in separation selectivity that occurs as mu(eff) of the less mobile enantiomer changes from cationic to anionic, a new feature, a separation selectivity maximum was predicted to occur in the resolving agent concentration range where both enantiomers migrate cationically provided that (i) K(RCD)/K(SCD) <1 and mu0(RCD)/mu0(SCD) >1 and (K(RCD)mu0(RCD))/(K(SCD)mu0(SCD)) > 1, or (ii) K(RCD)/K(SCD) >1 and mu0(RCD)/mu0(SCD) <1 and (K(RCD)mu0(RCD))/(K(SCD)mu0(SCD)) <1. This hitherto unseen separation selectivity pattern was experimentally verified during the nonaqueous capillary electrophoretic separation of the enantiomers of four weak base analytes in acidic methanol background electrolytes with octakis(2,3-diacetyl-6-sulfato)-gamma-cyclodextrin (ODAS-gammaCD) as resolving agent.


Waveguide damage detection by the matching pursuit approach employing the dispersion-based chirp functions.
If the wave mode used in guided wave non-destructive inspection is dispersive, reflected pulses from damaged parts may be significantly distorted due to wave dispersion. The main concern, in this case, is how to detect the reflected pulses in noisy signals, and to extract meaningful damage information from the detected pulses. However, current signal processing techniques used for guided wave inspection do not account for pulse dispersion, so the extracted information is often not so accurate. The objective of this study is to develop an efficient technique to deal with dispersed pulses for guided-wave nondestructive evaluation. Our idea is to model dispersed pulses by chirp functions of special form that can simulate up to quadratically varying group delay. To determine the parameters of the chirp functions approximating dispersed, reflected pulses, an adaptive matching pursuit algorithm is employed. Once the characterizing parameters are found, the damage location and extent can be estimated. The proposed method is tested with experimentally measured signals of longitudinal waves in a circular cylinder.


Minimal effect of CD103 expression on the control of a chronic antiviral immune response.
Impaired antiviral CD8 and CD4 T-cell responses are often associated with chronic viral infections. Cell-intrinsic as well as cell-extrinsic mechanisms are thought to dampen such responses, for example programmed death 1 receptor (PD-1) expression on T cells, and interleukin (IL)-10 production primarily by dendritic cells (DCs), have been shown to support viral persistence by suppressing immune responses. Here we demonstrate that CD103, an alpha E integrin necessary for T-cell homing and retention in the gut and other epithelia expressed by the majority of naïve CD8(+), and CD4(+)CD25(+) T cells and some DC subsets, is unnecessary for controlling T-cell responses during chronic lymphocytic choriomeningitis virus clone 13 (LCMV cl13) infection. T-cell analysis following viral infection showed that the primary as well as the memory CD8(+) and CD4(+) T-cell responses among CD103-sufficient and CD103-deficient mice were identical. In addition, no rescue of cytokine production by virus-specific T cells or alterations in viral titers in the absence of intrinsic CD103 expression was observed. Interestingly, CD103 levels on the effector CD8(+) T cells became reduced soon after virus infection, with a small proportion of cells co-expressing PD-1 and CD103. In contrast, although no substantial differences in the frequency and number of the CD4(+)CD25(+) cell population were seen, CD103 expression increased significantly over time in this population, correlating with viral persistence. Thus, a lack of CD103 expression does not affect functional impairment of effector T-cell responses during chronic viral infection.


Iron restriction inside macrophages regulates pulmonary host defense against Rhizopus species.
Mucormycosis is a life-threatening respiratory fungal infection predominantly caused by Rhizopus species. Mucormycosis has incompletely understood pathogenesis, particularly how abnormalities in iron metabolism compromise immune responses. Here we show how, as opposed to other filamentous fungi, Rhizopus spp. establish intracellular persistence inside alveolar macrophages (AMs). Mechanistically, lack of intracellular swelling of Rhizopus conidia results in surface retention of melanin, which induces phagosome maturation arrest through inhibition of LC3-associated phagocytosis. Intracellular inhibition of Rhizopus is an important effector mechanism, as infection of immunocompetent mice with swollen conidia, which evade phagocytosis, results in acute lethality. Concordantly, AM depletion markedly increases susceptibility to mucormycosis. Host and pathogen transcriptomics, iron supplementation studies, and genetic manipulation of iron assimilation of fungal pathways demonstrate that iron restriction inside macrophages regulates immunity against Rhizopus. Our findings shed light on the pathogenetic mechanisms of mucormycosis and reveal the role of macrophage-mediated nutritional immunity against filamentous fungi.


Sequence self-selection by cyclic phase separation.
The emergence of functional oligonucleotides on early Earth required a molecular selection mechanism to screen for specific sequences with prebiotic functions. Cyclic processes such as daily temperature oscillations were ubiquitous in this environment and could trigger oligonucleotide phase separation. Here, we propose sequence selection based on phase separation cycles realized through sedimentation in a system subjected to the feeding of oligonucleotides. Using theory and experiments with DNA, we show sequence-specific enrichment in the sedimented dense phase, in particular of short 22-mer DNA sequences. The underlying mechanism selects for complementarity, as it enriches sequences that tightly interact in the dense phase through base-pairing. Our mechanism also enables initially weakly biased pools to enhance their sequence bias or to replace the previously most abundant sequences as the cycles progress. Our findings provide an example of a selection mechanism that may have eased screening for auto-catalytic self-replicating oligonucleotides.


Recent advances in component resolved diagnosis in food allergy.
Due to the high prevalence of food allergic diseases globally there are increasing demands in clinical practice for managing IgE-mediated conditions. During the last decade, component resolved diagnostics has been introduced into the field of clinical allergology, providing information that cannot be obtained from extract-based tests. Component resolved data facilitate more precise diagnosis of allergic diseases and identify sensitizations attributable to cross-reactivity. Furthermore it assists risk assessment in clinical practice as sensitization to some allergenic molecules is related to persistence of clinical symptoms and systemic rather than local reactions. The information may also aid the clinician in prescription of oral immunotherapy (OIT) in patients with severe symptoms, and in giving advice on food allergen avoidance or on the need to perform food challenges. The use of allergen components is rapidly evolving and increases our possibility to treat food allergic patients with a more individual approach. Using molecular allergology, we can already now better diagnose, prognose and grade the food allergy. In summary, daily routine molecular allergy diagnostics offers a number of benefits that give us a higher diagnostic precision and allow for better management of the patient.


Expression and function of intestinal hexose transporters after small intestinal denervation.
The role of neural regulation in expression and function of intestinal hexose transporters is unknown. The aim of this study is to determine the role of intestinal innervation in gene expression and function of the membrane hexose transporters, SGLT1, GLUT2, and GLUT5 in the enterocyte. We hypothesize that denervation of the small intestine decreases expression of hexose transporters, which leads to decreased glucose absorption. Six groups of Lewis rats were studied (n = 6 each) as follows: control, 1 week after sham laparotomy, 1 and 8 weeks after syngeneic (no immune rejection) orthotopic small-bowel transplantation (SBT) (SBT1 and SBT8) to induce complete extrinsic denervation, and 1 and 8 weeks after selective disruption of intrinsic neural continuity to jejunoileum by gut transection and reanastomosis (T/A1 and T/A8). All tissue was harvested between 8 AM and 10 AM. In duodenum, jejunum, and ileum, mucosal messenger RNA (mRNA) levels were quantitated by real-time polymerase chain reaction (PCR), protein by Western blotting, and transporter-mediated glucose absorption using the everted sleeve technique. Across the 6 groups, the relative gene expression of hexose transporter mRNA and protein levels were unchanged, and no difference in transporter-mediated glucose uptake was evident in any region. The glucose transporter affinity (K(m)) and functional transporter levels (V(max)) calculated for duodenum and jejunum showed no difference among the 6 groups. Baseline regulation of hexose transporter function is not mediated tonically by intrinsic or extrinsic neural continuity to the jejunoileum.


Strategies for Improving Hepatitis C Treatment Access in the United States: State Officials Address High Drug Prices, Stigma, and Building Treatment Capacity.
Curative treatments for hepatitis C virus (HCV) can alter the course of a devastating epidemic, but high drug prices have contributed to restrictions on HCV treatment access. We aimed to learn how state health agencies have responded to the challenges of treatment access for HCV. Qualitative study using semistructured key informant interviews focused on aspects of HCV treatment access between June 2016 and March 2017. Content analysis was used to identify dominant themes. United States. Eighteen health officials and treatment advocates across 6 states selected using purposive sampling. Drug pricing is the most important barrier to access, encouraging restrictive authorization criteria from payers that in turn discourage providers from offering treatment. However, payers have not experienced the budget impact that was initially feared. Although authorization criteria are being lifted for fee-for-service Medicaid programs, ensuring that managed care organizations follow suit remains a challenge. The effect of stigma, a shortage of treating providers, and lack of political motivation are additional challenges to expanding treatment. The response to the human immunodeficiency virus epidemic can augment or inform strategies for HCV treatment delivery, but this is limited by the absence of dedicated funding. While treatment eligibility criteria for HCV treatment are improving, many other barriers remain to achieving the scale-up needed to end the epidemic. Political disinterest, stigma, and a lack of specialty providers are continued barriers in some jurisdictions. States may need to invest in strategies to overcome these barriers, such as engaging in public and provider education and ensuring that treatment by primary care providers is reimbursed. Despite uncertainty about how federal policy changes to Medicaid may affect states' ability to respond, states can identify opportunities to improve access.


Surveillance cultures following a regional outbreak of carbapenem-resistant Acinetobacter baumannii.
The primary aim of this study was to assess the epidemiology of carbapenem-resistant Acinetobacter baumannii (CRAB) for 9 months following a regional outbreak with this organism. We also aimed to determine the differential positivity rate from different body sites and characterize the longitudinal changes of surveillance test results among CRAB patients. Observational study. A 607-bed tertiary-care teaching hospital in Milwaukee, Wisconsin. Any patient admitted from postacute care facilities and any patient housed in the same inpatient unit as a positive CRAB patient. Participants underwent CRAB surveillance cultures from tracheostomy secretions, skin, and stool from December 5, 2018, to September 6, 2019. Cultures were performed using a validated, qualitative culture method, and final bacterial identification was performed using mass spectrometry. In total, 682 patients were tested for CRAB, of whom 16 (2.3%) were positive. Of the 16 CRAB-positive patients, 14 (87.5%) were residents from postacute care facilities and 11 (68.8%) were African American. Among positive patients, the positivity rates by body site were 38% (6 of 16) for tracheal aspirations, 56% (9 of 16) for skin, and 82% (13 of 16) for stool. Residents from postacute care facilities were more frequently colonized by CRAB than patients admitted from home. Stool had the highest yield for identification of CRAB.


PRR14 overexpression promotes cell growth, epithelial to mesenchymal transition and metastasis of colon cancer via the AKT pathway.
PRR14 (Proline rich protein 14) was firstly identified for its ability to specify and localize heterochromatin during cell cycle progression. Aberrant expression of PRR14 is associated with the tumorigenesis and progression of lung cancer. However, its involvement in colon cancer remains unknown. Herein, we report the role of PRR14 in colon cancer. Colon cancer tissue microarray was used to analyze and compare the expression of PRR14 among some clinicopathological characteristics of colon cancer. HCT116 and RKO cells were transfected with siRNA to downregulate PRR14 expression. The roles of PRR14 in proliferation, migration and invasion of the cell lines were determined using cell counting kit-8, colony formation assay, wound healing assay and transwell assays respectively. The expression of PRR14 was measured using immunofluorescence, qRT- PCR and western blot. Epithelial-mesenchymal transition (EMT) markers were determined by western blot. PRR14 was highly expressed in colon cancer tissues, and the expression level was correlated with tumor size, distant metastasis and Tumor Node Metastasis stages. Functional study revealed that downregulation of PRR14 inhibited colon cancer cells growth, migration and invasion. Furthermore, knockdown of PRR14 inhibited epithelial-mesenchymal transition (EMT) process, cell cycle-associated proteins expression and p-AKT level. PRR14 may promote the progression and metastasis of colon cancer, and may be a novel prognostic and therapeutic marker for the disease.


Institutional volume and the effect of recipient risk on short-term mortality after orthotopic heart transplant.
We developed a validated 50-point recipient risk index predicting short-term mortality after orthotopic heart transplant (OHT). This study examined the relationship between institutional volume and recipient risk on post-OHT mortality. We used United Network for Organ Sharing (UNOS) data to identify primary OHT recipients between January 2000 and April 2010. Centers were stratified by mean annual volume. Preoperative Index for Mortality Prediction After Cardiac Transplantation risk scores were calculated for each patient with our validated 50-point system. Primary outcomes were 30-day and 1-year survivals. Multivariable logistic regression analysis included interaction terms to examine effect modification of risk and volume on mortality. In all, 18,226 patients underwent transplant at 141 centers: 1173 (6.4%) recipients at low-volume centers (<7 procedures/y), 5353 (29.4%) at medium-volume centers (7-15 procedures/y), and 11,700 (64.2%) at high-volume centers (>15 procedures/y). Low center volume was associated with worse 1-year mortality (odds ratio, 1.58; 95% confidence interval, 1.30-1.92; P < .001). For 1-year survival, there was significant positive interaction between center volume and recipient risk score (odds ratio, 1.04; 95% confidence interval, 1.01-1.07; P = .02), indicating effect of risk on mortality at low-volume centers greater than from either variable analyzed individually. Among high-risk recipients (score ≥10), 1-year survival was improved at high-volume centers (high, 79%; medium, 75%; low, 64%). In analysis of UNOS data with our validated recipient risk index, institutional volume acted as an effect modifier on association between risk and mortality. High-risk patients had higher mortality at low-volume centers; differences dissipated among lower-risk recipients. These data support a mandate for high-risk transplants at higher-volume centers.


Pilgrims satisfaction with ambulatory health services in Makkah, 2008.
The main objective of this study was to assess the level and correlates of patients' satisfaction with ambulatory health services provided for pilgrims during Hajj period in 2008. This was a facility-based, cross-sectional study conducted in the Makkah region during the Hajj season in December 2008. A two-stage technique was used to select 500 patients from those who attended the ambulatory health services. One hundred subjects were selected by systematic random sampling (every fifth) from each of the five hospitals included in the study and asked to fill in a pilot-tested self-administered questionnaire. A total of 487 questionnaires were analyzed. Descriptive statistics and t-test, Mann Whitney test and ANOVA, or Kruskal-Wallis test was used as appropriate after checking for normality. Level of significance level was set to be <0.05 throughout the study. From 478 subjects analyzed, 390 (81.6%) were man, 345 (72.2%) were married, 28.9% had either intermediate or high secondary school education, and 2.4% were skilled laborers. The total satisfaction score for health facilities was 20.45 ± 4.03 of 25. The satisfaction scores were 20.15 ± 4.7 of 25 for patient satisfaction with physicians and 21.35 ± 4.5 for patient satisfaction with paramedical personnel. The overall satisfaction score was 61.5 ± 4.5 of 75 points. There were significant relations between total satisfaction of health facilities with education level and with occupation (P = 0.012, 0.001, respectively). The total satisfaction of patients with physicians was significant only with education level. The overall satisfaction score had a significant relation with occupation (P = 0.03), but a borderline relation with the education level (P = 0.056). Satisfaction with ambulatory Hajj health services is acceptable. Some physicians and waiting area services need special attention to improve satisfaction levels with ambulatory health in the subsequent Hajj seasons.


A detailed study of patent system for protection of inventions.
Creations of brain are called intellect. Since these creations have good commercial value, are called as property. Inventions are intellectual property and can be protected by patents provided the invention is novel, non-obvious, useful and enabled. To have fare trade among member countries, World Trade Organisation proposed TRIPS agreement. India had taken necessary initiation by signing the World Trade Organisation agreement and transformed to global needs. The aim of this article is to enlighten pharmaceutical professionals especially in the field of research and development about planning inventions by thorough review of prior-art, which saves time and money. A thorough understanding is made possible by providing details of origin; present governing bodies, their role along with the Act that is safeguarding the patent system.


Identification and engineering of human variable regions that allow expression of stable single-chain T cell receptors.
Single-chain antibody fragments (scFv), consisting of two linked variable regions (V(H) and V(L)), are a versatile format for engineering and as potential antigen-specific therapeutics. Although the analogous format for T cell receptors (TCRs), consisting of two linked V regions (Vα and Vβ; referred to here as scTv), could provide similar opportunities, all wild-type scTv proteins examined to date are unstable. This obstacle has prevented scTv fragments from being widely used for engineering or therapeutics. To further explore whether some stable human scTv fragments could be expressed, we used a yeast system in which display of properly folded domains correlates with ability to express the folded scTv in soluble form. We discovered that, unexpectedly, scTv fragments that contained the human Vα2 region (IMGT: TRAV12 family) were displayed and properly associated with different Vβ regions. Furthermore, a single polymorphic residue (Ser(α49)) in the framework region conferred additional thermal stability. These stabilized Vα2-containing scTv fragments could be expressed at high levels in Escherichia coli, and used to stain target cells that expressed the specific pep-HLA-A2 complexes. Thus, the scTv fragments can serve as a platform for engineering TCRs with diverse specificities, and possibly for therapeutic or diagnostic applications.


Thalidomide Improves Psoriasis-like Lesions and Inhibits Cutaneous VEGF Expression without Alteration of Microvessel Density in Imiquimod- induced Psoriatic Mouse Model.
Psoriasis is a chronic inflammatory skin disorder of unknown etiology. Increasing evidence suggests that psoriasis is probably an angiogenesis-dependent disease. Thalidomide has been reported being able to inhibit the effects of fibroblast growth factor 2 and vascular endothelial growth factor (VEGF), and inhibit tumour necrosis factor-alpha synthesis, and suppress tumour necrosis factor-induced nuclear factor-kappa B activation in Jurkat cells, resulting in suppression of proliferation inflammation, angiogenesis, and the immune system, which are related to the pathogenesis of psoriasis. Our study evaluated the influence of thalidomide on the lesional alterations, VEGF expressions and angiogenesis in imiquimod-induced mouse model. Balb/c female mice (n=48) 8-12 weeks of age were randomly divided into 6 groups including negative control (vaseline cream), positive control (5% imiquimod cream), and experimental groups including low-dose (10 mg/kg.d), moderate-dose (30 mg/kg.d) and high-dose thalidomide (85 mg/kg.d), and acitretin group (6 mg/kg.d). Serum levels of VEGF-A were quantified by enzyme-linked immunosorbent assay. VEGF protein expression was measured by western blotting and the microvessel density by immunohistochemical staining. The total psoriasis area and severity index scores in the moderate- and high-dose thalidomide and acitretin groups decreased significantly (p<0.001 for each), and so were the total Baker's scores in the high-dose thalidomide (p=0.008) and acitretin groups (p=0.021). The mean thickness of the epidermis in the experimental and acitretin groups decreased significantly, respectively (p<0.001 for all); the acitretin group was the thinnest. The cutaneous VEGF protein levels down-expressed significantly in the moderate- and high-dose thalidomide groups (p<0.05 for both), while those in the low-dose thalidomide and acitretin did not (p>0.05 for both). There were no differences for serum VEGF-A levels and the density of microvessels among the positive and experimental groups. Thalidomide can improve the psoriasis-like lesions and inhibit the expression of cutaneous VEGF in imiquimod-induced psoriatic model with dose-dependence, however, it does not alter circulating VEGF-A levels and microvessel density in dermis.


Myocardial conditioning: opportunities for clinical translation.
Myocardial conditioning is an endogenous cardioprotective phenomenon that profoundly limits infarct size in experimental models. The current challenge is to translate this paradigm from the laboratory to the clinic. Accordingly, our goal in this review is to provide a critical summary of the progress toward, opportunities for, and caveats to, the successful clinical translation of postconditioning and remote conditioning, the 2 conditioning strategies considered to have the broadest applicability for real-world patient care. In the majority of phase II studies published to date, postconditioning evoked a ≈35% reduction of infarct size in ST-segment-elevation myocardial infarction patients. Essential criteria for the successful implementation of postconditioning include the appropriate choice of patients (ie, those with large risk regions and negligible collateral flow), timely application of the postconditioning stimulus (immediately on reperfusion), together with proper choice of end points (infarct size, with concomitant assessment of risk region). Remote conditioning has been applied in planned ischemic events (including cardiac surgery and elective percutaneous coronary intervention) and in ST-segment-elevation myocardial infarction patients during hospital transport. Controversies with regard to efficacy have emerged, particularly among surgical trials. These disparate outcomes in all likelihood reflect the remarkable heterogeneity within and among studies, together with a deficit in our understanding of the impact of these variations on the infarct-sparing effect of remote conditioning. Ongoing phase III trials will provide critical insight into the future role of postconditioning and remote conditioning as clinically relevant cardioprotective strategies.


Transcription regulation and animal diversity.
Whole-genome sequence assemblies are now available for seven different animals, including nematode worms, mice and humans. Comparative genome analyses reveal a surprising constancy in genetic content: vertebrate genomes have only about twice the number of genes that invertebrate genomes have, and the increase is primarily due to the duplication of existing genes rather than the invention of new ones. How, then, has evolutionary diversity arisen? Emerging evidence suggests that organismal complexity arises from progressively more elaborate regulation of gene expression.


Image identification and restoration in the subband domain.
When faced with a large support point spread function (PSF), the iterative expectation-maximization (EM) algorithm, which is often used for PSF identification, is very sensitive to the initial PSF estimate. To deal with this problem, the authors propose to do EM image identification and restoration in the subband domain. After the image is first divided into subbands, the EM algorithm is applied to each subband separately. Since the PSF can be taken to have smaller support in each subband, these subbands should be less of a problem with the EM model identification. They also introduce an adaptive subband EM method for use in the upper frequency subbands.


Efficacy and safety of intravenous meropenem and tobramycin versus ceftazidime and tobramycin in cystic fibrosis.
Cystic fibrosis (CF) is characterized by chronic bacterial broncho-pulmonary infection. Although intravenous (i.v.) antibiotic therapy is regarded as standard treatment in CF, only few randomised trials comparing different antibiotic compounds exist. We report on a prospective multicenter interventional trial of i.v. meropenem (120 mg/kg/day) or i.v. ceftazidime (200-400 mg/kg/day), each administered together with i.v. tobramycin (9-12 mg/kg/day). Outcome measures were changes in lung function, microbiological sputum burden and blood inflammatory marker. Liver and renal function values were measured to assess safety. One hundred eighteen patients (59/59) were included into the study with the following indications: first infection of P. aeruginosa (n=6), acute pulmonary exacerbation (n=34) and suppression therapy of chronic P. aeruginosa colonization (n=78). Both treatments improved lung function measures, bacterial sputum burden and CRP levels with no differences between treatment groups observed. A significant higher elevation for alkaline phosphatase (p<0.0001) was observed for patients in the meropenem/tobramycin group. i.v. antibiotic therapy in CF patients with meropenem/tobramycin is as effective as with ceftazidime/tobramycin regarding lung function, microbiological sputum burden and systemic inflammatory status. Hepato-biliary function should be monitored carefully during i.v. treatment, possibly important in CF patients with pre-existing liver disease.


Hyperemesis gravidarum and the risk of childhood cancer - A case-control study in Denmark.
Only a few studies have reported on the association between hyperemesis gravidarum and the risk of childhood cancer. We examined possible associations in this population-based study in Denmark. Pediatric cancer cases (n = 6420) were ascertained from the Denmark Cancer Registry among children born between 1977 and 2013. Twenty-five controls were matched to each case by sex and birth date from the Central Person Registry (n = 160500). Mothers with hyperemesis gravidarum were ascertained from the National Patient Register. The risk of childhood cancer was estimated using conditional logistic regression. In a separate analysis, we examined pregnancy prescription of antinauseant medications, ascertained from the National Pharmaceutical Register, to determine associations with childhood cancers. In Denmark, hyperemesis gravidarum was associated with an increased risk of childhood cancer [all types combined; Odds Ratio (OR) = 1.43, 95% confidence interval (CI) 1.12, 1.81; n = 73 exposed cases). Hyperemesis gravidarum was also associated with an increased risk of neuroblastoma (OR = 2.52, 95% CI 1.00, 6.36; n = 5 exposed cases), acute lymphoblastic leukemia (OR = 1.63, 95% CI 0.98, 2.72; n = 16 exposed cases), and non-Hodgkin's lymphoma (OR = 2.41, 95% CI 0.95, 6.08; n = 5 exposed cases). We observed no childhood cancer risk increase from antinauseant prescriptions (OR = 1.05, 95% CI 0.84, 1.30; n = 91 exposed cases). Our results are suggestive of an association between hyperemesis gravidarum and the overall cancer risk in offspring, particularly for neuroblastoma. Mothers with hyperemesis gravidarum should be closely monitored and receive appropriate treatment during pregnancy.


In vivo mutational analysis of the NGFI-A zinc fingers.
NGFI-A is a mammalian transcription factor that contains zinc fingers similar to those observed in several other proteins, including NGFI-C and Krox-20. To define precisely the DNA binding domain of NGFI-A, we selected mutants using a chimeric transcriptional activator that contains the NGFI-A zinc finger domain sandwiched between the lexA DNA binding domain and the GAL4 transcriptional activating domain. Expression of this lexA-NGFI-A-GAL4 (LAG) trimeric protein in yeast significantly retarded their growth, unlike an activator containing only the lexA and GAL4 components. This suggested that LAG inappropriately regulates genes in yeast that contain NGFI-A binding sites. Yeast that contained LAG reverted to wild-type growth at high frequency by inactivation of LAG. The mutations recovered from these revertants were specifically limited to the 83-residue NGFI-A zinc finger domain by requiring that the lexA and GAL4 portions of the LAG chimera remain functional. Nearly all of the 93 mutants obtained contained single missense mutations that mapped within the zinc fingers to residues thought to be important for zinc finger function. Deletion analysis of native NGFI-A verified that residues distant from the zinc fingers do not influence DNA binding, thus establishing the minimal functional DNA binding domain. Interestingly, many zinc finger residues ascribed specific functions by x-ray crystallography were never mutated in yeast, implying that the identity of these residues is not critical. Surprisingly, not all of the mutations tested significantly impaired NGFI-A-specific DNA binding, suggesting that the function of these zinc fingers is more diverse than previously recognized.


Effects of transgenic Bacillus thuringiensis maize grain on B. thuringiensis-susceptible Plodia interpunctella (Lepidoptera: Pyralidae).
Percentage survivorship, developmental time, adult body length, and sex ratio of Plodia interpunctella (Hübner) reared on field-produced grain from sixteen cultivars of maize, Zea mays L., including several transgenic Bacillus thuringiensis (Bt) Berliner hybrids and selected non-Bt isolines, were evaluated under laboratory conditions. Compared with isolines, development was delayed and survivorship reduced for P. interpunctella reared on grain from transgenic hybrids with the CaMV/35s promoter that express Cry1Ab protein. Similarly, compared with non-Bt hybrids, a transgenic hybrid with the CaMV/35s promoter that expresses Cry9C protein delayed development, decreased survivorship, and caused reductions in adult body length of P. interpunctella. In contrast, no significant differences in P. interpunctella developmental times or survivorship were observed between transgenic hybrids with the PEPC promoter expressing Cry1Ab and their isolines. Additionally, developmental time, survivorship, and adult body length were similar between P. interpunctella reared on a transgenic hybrid with the CaMV/35s promoter expressing Cry1Ac and non-Bt hybrids. Our data demonstrate that transgenic Bt maize grain, especially grain from hybrids with the CaMV/35s promoter expressing Cry1Ab or Cry9C, can significantly affect B. thuringiensis-susceptible P. interpunctella populations up to 4 or 5 mo after harvest.


Regulation of nonsmall-cell lung cancer stem cell like cells by neurotransmitters and opioid peptides.
Nonsmall-cell lung cancer (NSCLC) is the leading type of lung cancer and has a poor prognosis. We have shown that chronic stress promoted NSCLC xenografts in mice via stress neurotransmitter-activated cAMP signaling downstream of beta-adrenergic receptors and incidental beta-blocker therapy was reported to improve clinical outcomes in NSCLC patients. These findings suggest that psychological stress promotes NSCLC whereas pharmacologically or psychologically induced decreases in cAMP may inhibit NSCLC. Cancer stem cells are thought to drive the development, progression and resistance to therapy of NSCLC. However, their potential regulation by stress neurotransmitters has not been investigated. In the current study, epinephrine increased the number of cancer stem cell like cells (CSCs) from three NSCLC cell lines in spheroid formation assays while enhancing intracellular cAMP and the stem cell markers sonic hedgehog (SHH), aldehyde dehydrogenase-1 (ALDH-1) and Gli1, effects reversed by GABA or dynorphin B via Gαi -mediated inhibition of cAMP formation. The growth of NSCLC xenografts in a mouse model of stress reduction was significantly reduced as compared with mice maintained under standard conditions. Stress reduction reduced serum levels of corticosterone, norepinephrine and epinephrine while the inhibitory neurotransmitter γ-aminobutyric acid (GABA) and opioid peptides increased. Stress reduction significantly reduced cAMP, VEGF, p-ERK, p-AKT, p-CREB, p-SRc, SHH, ALDH-1 and Gli1 in xenograft tissues whereas cleaved caspase-3 and p53 were induced. We conclude that stress neurotransmitters activate CSCs in NSCLC via multiple cAMP-mediated pathways and that pharmacologically or psychologically induced decreases in cAMP signaling may improve clinical outcomes in NSCLC patients.


Increased Understanding of Stem Cell Behavior in Neurodegenerative and Neuromuscular Disorders by Use of Noninvasive Cell Imaging.
Numerous neurodegenerative and neuromuscular disorders are associated with cell-specific depletion in the human body. This imbalance in tissue homeostasis is in healthy individuals repaired by the presence of endogenous stem cells that can replace the lost cell type. However, in most disorders, a genetic origin or limited presence or exhaustion of stem cells impairs correct cell replacement. During the last 30 years, methods to readily isolate and expand stem cells have been developed and this resulted in a major change in the regenerative medicine field as it generates sufficient amount of cells for human transplantation applications. Furthermore, stem cells have been shown to release cytokines with beneficial effects for several diseases. At present however, clinical stem cell transplantations studies are struggling to demonstrate clinical efficacy despite promising preclinical results. Therefore, to allow stem cell therapy to achieve its full potential, more insight in their in vivo behavior has to be achieved. Different methods to noninvasively monitor these cells have been developed and are discussed. In some cases, stem cell monitoring even reached the clinical setting. We anticipate that by further exploring these imaging possibilities and unraveling their in vivo behavior further improvement in stem cell transplantations will be achieved.


β-Catenin activation and illicit receptor expression in adrenocortical cells.
Adrenal cortisol-producing tumors can express illicit membrane receptors such as luteinizing hormone (LH), glucose-dependent insulinotropic peptide (GIP) or type 4 and 7 serotonin (5-HT4/7) receptors. Abnormal expression of the LH receptor (LH-R) has been ascribed to the activation of the Wnt/β-catenin signaling pathway in adrenocortical cells. In the present study, we have investigated whether β-catenin activation may also trigger the illegitimate expression of GIP and 5-HT receptors. Three models of β-catenin activation in adrenocortical cells were used: an APC-mutated adrenocortical tumor, human-transfected adrenocortical cells and genetically modified mouse adrenal glands. The methods employed include quantitative reverse transcription PCR, immunohistochemistry and measurement of cortisol secretion by cultured tumor cells. Abnormal expression of the GIP, 5-HT7and LH receptors was observed in the APC-mutated adrenocortical tumor tissue. In addition, GIP, 5-HT and human chorionic gonadotropin stimulated cortisol production from tumor cells in primary culture. Conversely, only the LHCGR was upregulated in human and mouse adrenocortical cells harboring the activation of β-catenin. Moreover, LH-R immunoreactivity was detected in clusters of zona fasciculata cells in the β-catenin-activated mouse model. Our data indicate that activation of the β-catenin signaling pathway can promote the illicit expression of functional LH-Rs in adrenal zona fasciculata cells but does not favor the abnormal expression of GIP and 5-HT receptors.


Optimization of the thermal conditions for processing hatchery waste eggs as meal for feed.
The purpose of this study was to optimize the thermal conditions for processing hatchery waste eggs (HWE) into rich feedstuff with lower electricity consumption by using response surface methodology. In the study, the effects of processing temperature and time on HWE meal (HWEM) quality and production were evaluated. As the results indicate, optimization was obtained when the processing lasted for 23 h at the fixed temperature of 65°C, resulting in higher protein digestibility in vitro (89.6%) and DM (88.5%) content of HWEM with lower electricity consumption (82.4 kWh/60 kg of HWE). No significant differences existed between the quality values predicted by mathematical formulae and those obtained through practical analyses in DM (87 vs. 88.5%), CP (39.2 vs. 38.3%), protein digestibility in vitro (90.7 vs. 89.6%), and electricity consumed (80.8 vs. 82.4 kWh/60 kg of HWE). Furthermore, the product derived from the optimized processing conditions had better biosecurity; Salmonella spp. were not found and Escherichia coli levels were substantially reduced (from 10(7) to 10(4) cfu/g). In summary, HWEM of superior quality can be produced when the processing conditions optimized in the current research are utilized.


Fetal liver position and perinatal outcome for congenital diaphragmatic hernia.
Despite advances in postnatal care, patients born with a congenital diaphragmatic hernia (CDH) suffer substantial morbidity and mortality. The present study was undertaken to determine the prognostic influence of prenatally-diagnosed liver herniation in the hemithorax in fetuses with CDH. The medical records of 48 patients evaluated for a prenatally-diagnosed left CDH were retrospectively reviewed. Patients were analysed according to the position of the liver by prenatal ultrasound; 32 fetuses had a major portion of the liver herniated into the left hemithorax ('liver up') and 16 had an intra-abdominal liver ('liver down'). Liver position was determined using colour-flow Doppler ultrasonography. There were two fetal deaths in the liver-up group and one in the liver-down group. The liver-up group more frequently required extracorporeal membrane oxygenation (ECMO) support (53 per cent) compared with the liver-down group (19 per cent). Postnatal survival was significantly less in the liver-up group (43 per cent) vs. the liver-down group (93 per cent). Fetuses with congenital diaphragmatic hernia and liver herniated into the hemithorax have a much worse prognosis than similarly afflicted fetuses without liver herniation. Prenatal ultrasonographic diagnosis of congenital diaphragmatic hernia allows for preparation for a critically ill newborn and aids in prenatal family counselling.